Proceedings of the international session of the 26th annual meeting October 18–20, 1984—Chiba,...

GastroenterologiaJaponica Vol. 20, No. 3 Copyright �9 1985 by The Japanese Society of Gastroenterology Printed in Japan

P R O C E E D I N G S O F T H E I N T E R N A T I O N A L S E S S I O N O F

T H E 2 6 T H A N N U A L M E E T I N G

October 18-20, 1 9 8 4 - - C h i b a , Japan

C h a i r m a n : Kun io O K U D A , M . D . , Ph .D .

Modera to r s :

International symposium (1):

Hepatitis vaccine R . P . B E A S L E Y ( T a i w a n ) a n d Tosh io S H I K A T A (Japan)

Cl in ica l t r ial of HB vaccine in the families

of HBV carriers

Y. KATO, H. MIYAMORI, M. UNOURA,

N. OHMIZO, N. TANAKA, K. KOBAYASHI

and N. H A T T O R I

1st Dept Intern Med, Sch Med,

Kanazawa Univ

An HB vaccine was given to 75 subjects and anti-

body formation in response to vaccination was

studied. They consisted of hospital workers and

families of HBeAg positive carriers. The subjects

younger than 10 years were given 10 pg of vaccine

and those who were 10 years or older 20 pg. The

second and third injections were given 4 and 24

weeks after the first. Twenty-eight weeks after the

first vaccination, anti-HBs was detected in 70 out of

the 75 subjects (93%). The anti-HBs response was

more marked in females and in younger individuals,

but the differences between males and females and

between the younger and older groups were not sig-

nificant. In subjects under 10 years of age, an ade-

quate anti-HBs response was obtained with 10 pg of

HBsAg protein. No clear correlation could be eluci-

dated between anti-HBs response and HLA because

even the siblings who showed a same low anti-HBs

response did not have one HLA haplotype in com-

mon, and a difference was observed in anti-HBs re-

sponse even among siblings who had the same two HLA haplotype.

A cl inical t r ial of hepati t is B vaccine in hospital employees

Y. ARAKAWA, S. AMAKI and T. SHIKATA

Depts Med and Pathol, Nihon Univ

Sch Med, Tokyo, Japan

We have carried out a clinical trial of hepatitis B

vaccine prepared in Japan. The study subjects con-

sisted of HBsAg-negative 205 hospital employees.

All recipients received subcutaneously three injec-

tions of HB vaccine at 0, 4 and 24 (or 20) weeks, at

three different dose levels of 16/./g, 40 pg and 20/./g

in Phase I, II and III studies, respectively. Ten per-

sons, who were positive for HBsAb before vaccina-

tion, showed an anamnestic response. Of the re-

maining HBsAb-negative 195 recipients, 179 de-

veloped a primary HBsAb response (82/95 in males

and 97/100 in females) and the other 16 failed to re-

spond to the vaccine. The higher immune response

rate was confirmed in recipients aged less than 50,

but a sexual difference in immune response was not

significant. None of the vaccinated persons showed

any sign of hepatic injury. The incidence of local or

general side effects was low. Peripheral T cell subsets

showed no significant difference between good and

weak responders to the vaccine. The overall rate of

HBsAb-positive vaccine recipients remained almost

unchanged during the 15-month follow-up period.

However, several vaccinees who lost the antibody 30

months later, were among the weak responders. The

HB vaccine was suggested to be safe an efficacious

when tested in a high risk population.

Hepatitis B vaccine trials in Korea

Bang-Hyun LIU

274 Proceedings of26th Annual Meeting Vol. 20, No. 3

Dept Med, Pusan Natl Univ Hosp, Pusan, Korea

Chronic HBV carr ier is es t imated more than three

& hal f mill ion in Korea and a round 8 - 1 0 % of these

carriers will be chronic hepatopathies . Since the

licensure of an inact ivated hepati t is B vaccine, H-B

Vax is r ecommended for immuniza t ion of infants,

chi ldren & adults who are exposed to h igh risk of he-

patit is B imfection.

H-B Vax (Merck) was in t roduced in Korea in July

1982 as a commercia l base. Subseqhently Pasteur

Vaccine in Aug. 1982, and currently Hepavax B,

Green Cross Co., Korea in Oct. 1983 are also avail-

able here and this t ime HBV vaccinat ion is widely

administered.

The authors studied on immune response of

heal thy adults to HB vaccine at the six months after

3 times injection and they also observed the i m m u n e

response in neonates to HB vaccine & HBIG.

Fur ther follow up studies on ant ibody response

after HB vaccinat ion will be carr ied out in future in

addi t ion to this pre l iminary report . And fur ther

prevention of hepati t is B infection will be expected

by active vaccinat ion p rog ram at highly contami-

na ted areas.

Efficacy of HBIG and HB vaccine i n

preventing perinatal transmission of HB virus to children born

to HBe antigen-positive carrier mothers

K. KANAI, S. M O R I O K A , M. NISHIDA,

H. N O T O , K. T A K A H A S H I and

K. M A T S U S H I T A

Depts Med, Pediatr, Obst and Gynecol and Epidemiol, Hamamatsu

Univ Sch Med, Shizuoka

The efficacy of the combina t ion of ant ihepat i t i s B

immunoglobu l in (HBIG) and HB vaccine for the

prevent ion of verical t ransmission of HB virus to the

babies born to eAg-positive HB carrier mothers has

been investigated. Babies received 150 m g of

H B I G . F (ab')~ i.v. within 2 hours of delivery and

serum anti-HBs ant ibody (HBsAb) titers were main-

ta ined by s.c. inject ion of 1 ml each of HBIG at 1

day, 2 and 5 mo after bir th , Three dose of HB vac-

cine conta in ing 32/~g HBsAg proteins was s tar ted at

3 mo (6 mo in some cases) after delivery at 1 mo

intervals. It has been disclosed that 58 of 59 babies

who are enrolled in this trial and older than 12 mo

of age, were actively immunized against HBsAg.

Vaccine which contains a lum adjuvant was found to

be more effective in produc ing HBsAb than the vac-

cine without adjuvant . Serum HBsAb titers were

well ma in ta ined in most babies who were actively

immunized without the addi t ional adminis t ra t ion of

vaccine. Mean serum HBsAb titers (PHA) 3, 6, 12

and 18 mo after ob ta in ing active immuniza t ion was

4.4, 4.3, 3.9 and 4.2, respectively. None of t h e

babies t reated by HBIG and HB vaccine revealed

positive HBcAb in their sera at the age of 12 mo. No

side effect was observed.

Immunoprophylaxis on HBV perinatal infection

R. Palmer BEASLEY and Lu-Yu H W A N G

15% to 20% of popula t ion in Ta iwan are HBV

carriers. The infections most occur in infants and

chi ldren often from their own monthers . W h e n the

mothe r is e ant igen positive (40 % of carriers) at least

90% of infants become carriers from the per ina ta l

infection. Passive prophylaxis with Hepati t is B Im-

mune Globul in (HBIG) only can prevent 75% of them from carriers. However, HB vaccine has shown

immunogenic in adults and children. For determin-

ing the immunogenic i ty and efficacy of HB vaccine (Merck Sharp & Dohme) against per inata l transmis-

sion in newborns, the clinical trial was conducted

since 1981. It demons t ra ted 100% i m m u n e response to 3 doses vaccine (20/~g) in 38 newborns. Efficacy

was a randomized trial for prevention of infants

born to e ant igen positive carr ier mothers . Since

1981 more t han 50,000 p regnan t women were

screened for HBsAg in Taipei . There are 260 infants

randomized to receive HBIG plus vaccine or HB

vaccine alone. Among the infants receiving HBIG

plus vaccine only 6% developed HBsAg carriers,

compared with 88% a m o n g the controls. A m o n g those infants who received HB vaccine but no

HBIG, 23% became HBsAg carriers. All the re-

ma in ing infants who d idn ' t become carriers develop antibodies.

June 1985 Proceedings of 26th Annual Meeting 275

The efficacy of HBIG in combination with vaccine

(94%) was superior to HBIG alone (71%) or vaccine

alone (75%). HBIG should be given immediately

after birth. To maximize compliance and minimize

costs, we recommend that HBV vaccination be ini-

tiated in the first week of life. For further evaluation

of the imrnunogenicity and efficacy of lower dose

vaccine, we have recruited 500 infants. The prelimi-

nary result will be known soon.

Efficacy of HB vaccine on HBV maternal i n fan t transmission

M. YANO, M. SHIMA, E. TSUNODA,

M. KOGA and T. M I N A M I N O

Inst Clin Res, Nagasaki Chuo Natl Hosp, Nagasaki, Japan

Prevention for HBV maternal infant transmission

is a very important treatment, especially in Asia

countries. 944 babies born to e antigen positive

mothers have been treated with passive or passive-

active immunization by our group study. All babies

were administrated HBIG just after birth and 1 or 2

additional HBIGs in every 3 months. We have ob-

served the efficacy of HB vaccine in 3 different sche-

dules. 171 babies are treated with schedule A, this is

a schedule of treatment with HBIG only. 108 babies

are treated with schedule B. In this schedule, HBIGs

are given 3 times, just after birth, 3 months and 6

months of age. HB vaccine is started at 6 months of

age and administrated 3 times. 665 babies are

treated with schedule C. In this schedule, HBIG is

given just after birth and 3 months of age. HB vac-

cine is given at 3 months, 4 months and 6 months of

age. The carrier rates of group A, B and C are

12.3%, 7.4% and 5.0%. 37 out of 944 babies

(3.9%) developed carrier state until 6 months of

age. There is no difference in the HBV carrier rates

of each group in this period. In the babies between 6

to 12 months of age, the HBV carrier rates of group

A, B and C are 3.4%, 4.9% and 1.2%. In the

babies of over 12 months of age, the carrier rates of

group A, B and C are 4 .5%, 0% and 0.7%. Preven-

tive efficacy with passive-active immunization is

better than with passive immunizat ion only. The de-

velopment of HBV carrier was restricted in the

babies who received HB vaccine around 3 month of

age compered with the babies who received after 6

months of age.

In ternat iona l s y m p o s i u m (2):

H e p a t i t i s v i r u s a n d h e p a t o c e l l u l a r carc inoma

Modera to r s : J . S U M M E R S (USA) a n d Masao O M A T A (Japan)

Clinico-epidemiological study of HBV carrier state

T. TSUJI, K. KAGAWA, T. OKUNO

and T. T A K I N O

3rd Dept Med, Kyoto Pref Univ Med, Kyoto, Japan

The aims were to investigate the prognosis, trans-

mission and annual incidence of carrier state. We

have done this study at HBV infectious district with

the most highest incidence (19%) in Japan for 10

years since 1974, where was a fishing village at the

coast of the Sea of Japan in Kyoto prefecture. There

were living about 300 people, who were detected an-

nual mass examination, and especially objected

same 103 people of them continuously. S-ALT and

others as liver function test examined, and HBsAg in

serum by R-PHA & RIA, anti-HBs by PHA & RIA,

HBeAg & anti-HBe by RIA, and anti-HBc by IAHA

& RIA. AFP & CEA used RIA. Uhrasonographic

method was done as the screening of HCC for 2 years

recently.

Results: 1) Carrier state was decreased annually

from 3rd year of the most highest levels (25.2%).

Therefore the incidence in 1983 was 18.3%. This

tendency was noticed female. 2) Some of carrier

states disappeared naturally, who was 17.1% and

this tendency also was female. Natural seroconver-

sion of HBsAg revealed that the incidence was

50.0%. They had all anti-HBe and the mean age

276 Proceedings of26th AnnualMeeting Vol. 20, No. 3

was 50.5 years old. The time interval took in 3 to 4

years. 3) HCC confirmed 2 cases of carrier states, in

whom were male and female each, and the inci-

dence of all candidates (232) was 0.86%, in which

was 0.9% of males and 0.8% of females. The inci-

dence of all carrier states (45) was 4.4%, in which

was 4.3% of males and 4 ,5% of females. These re-

sults were suggested that HBV could be had some

behavior of the onchogenesity in liver cell.

A prospective study on the development of hepatocellular carcinoma from

liver cirrhosis

Takeshi K U R I H A R A and Hiroshi OBATA

Inst Gastroenterol, Tokyo Women's Med Coll

We made a prospective study on the development

of hepatocellular carcinoma (HCC) in patients with

liver cirrhosis.

Among the patients who were diagnosed as liver

cirrhosis during the period from April 1970 until

May 1977, 110 patients, on whom we were able to

continue follow-up observations until May 1984,

were used as the subjects of the present study. Those

patients were classified into three groups by the anti-

HBc titer (IAHA Method) to carry out the investiga-

tion. Group I included those who were continuously

infected with HBV (anti-HBc titer~210), Group II,

those who has the infection in the past (anti-HBc

titer<29), and Group III, those who did not have

the infection.

HCC developed in 33 cases (30.0%) which were

further broken down as follows: 10 out of 33 cases of

Group I, 13 out of 35 cases of Group II, and 10 out

of 42 cases of Group III. The average observation

period until the development of HCC was 3 years

and 9 months in Group I, 5 years and 5 months in

Group II, and 5 years and 11 months in Group III,

In Group I, HCC developed at the comparatively

early stage and a decreasing tendency was observed

in those cases that were observed for a longer period

of time. On the other hand, in the non-infected

group, there were many cases in which development

of HCC occurred after a long period of observation,

indicating a difference between the two groups.

Hepatocellular carcinoma and HBV infection

R. Palmer BEASLEY

Hepatocellular carcinoma (HCC) is one of the commonest cancers in the world and chronic HBV virus infection is the usual underlying event preceed- ing its occurrence. Throughout the world most cases occur among persons with serological markers of

HBV infection, most of whom are HBsAg carriers; only a small proportion show no evidence of prior HBV infection. Even though there is striking world- wide geographic variation in the incidence of HCC,

there is a close international correlation with the prevalence of HBsAg carriers. Many HBV carriers

in the world were probably infected in early life by their own carrier mothers, and mother to child transmission appears to have unique importance in relation to HCC.

In a long term followup study begun in 1975 of 22,707 middleage men in Taiwan, during which there has been an average of 6.2 years of followup per man. 116 cases of HCC have occurred: 113 among 3,454 HBsAg carriers, and 3 among 19,253 non-carriers, a relative risk of 217. The incidence was 528/100,000/year among carriers, and the estimated life-time risk of death from HCC and /o r cirrhosis is approximately 40 %.

The risk of HCC among HBV carriers in the Taiwan prospective study was related to the presence of the following factors among the study subjects when they entered the study: 1) clinical cirrho-

sis, 2) history of hepatitis, 3) elevated serum anti- HBc IgM, 4) e antigen serum positivity. Even

though HBV is the major cause of HCC, it is probably not the only one. Other factors, especially aflatoxins, probably cause HCC and there may also be causitive interaction.

T h e role of HBV in the development of hepatocellular carc inoma

K. OKITA, T. KONISHI and Y. KADO

Dept Med, Yamaguchi Univ, Ube, Yamaguchi

We have performed the following clinico-patho-

logical studies for elucidation of possible role of


HBV-in hepatocarcinogenesis. Since 1970, 206 cases

of HCC have been collected and 45.2% of them

were positive for HBV-infection in the past. How-

ever, regarding juvenile case of HCC ranging from

11 to 39 years old, 10 out of 12 cases (83.3%)

showed positive for HBsAg or HBsAb. Integration of

HBV-DNA into cancer cell DNA was assayed using

surgically resected materials and was observed in all

cases who showed HBV associated markers in the

sera. These evidences suggest the mode of integra-

tion of HBV-DNA in host cell DNA must be impor-

tant for elucidation of close relationship between

HBV-infection and cancer development. In addi-

tion, in 14 cases who were followed over 3 years

through preneoplastic state, the validities of para-

meters for high risk group of HCC and preneoplastic

lesions directly related to cancer-development which

we already reported were estimated again.

Integration of hepati t is B virus DNA in neoplastic and non-neoplastic

liver diseases

O. YOKOSUKA, M. OMATA, F. IMAZEKI,

M. RYUU and K. OKUDA

1st Dept Med and 2nd Dept Surg, Chiba

Univ Sch Med, Chiba, Japan

By Southern blot hybridization technique using

~2P-labeled HBV DNA, integration of HBV DNA in

liver tissue was studied in 63 cases of non-neoplastic

chronic liver diseases (33 CPH, 19 CAH, 11 cirrho-

sis). Various forms of replicative free viral DNAs

were found in 34 of 38 (89%) of HBeAg positive, in

4 of 18 (22%) of anti-HBe positive, and in 2 of 7

(29%) both negative cases. High molecular weight

radioactive signals on electrophoresis were found in

4 of 63 (6%) cases of non-neoplastic liver diseases.

Two such patients had discrete bands of HBV DNA

after EcoR I and Hind III digestions.

Similarly, integration of HBV DNA was studied

in 14 cases of hepatocellular carcinoma (HCC).

Integrated HBV DNA was detected in all 4 HBsAg

seropositive and 3 of 10 (30%) seronegative cases.

Among these 3 HBsAg seronegative HCC cases who

had integrated HBV DNA, one had been thought to

be a sequla to non-A non-B post-transfusion chronic

liver disease, and another alcoholic liver disease by

conventional examinations.

Conclusion: In non-neoplastic liver diseases, pres-

ence of HBeAg in serum indicates frequent and

large quantities of free viral DNA in liver tissue, but

the presence of anti-HBe, believed to indicate ab-

sence of infectivity, does not necessarily exclude it.

By the application of molecular biology techniques,

new fields have been developed in the study of the

role of HB virus in hepatocarcinogenesis.

State of hepatit is B virus DNA in liver and hepatocellular carcinoma

tissues of HBV carriers

Keiji M I T A M U R A

Inst Clin Med, Univ of Tukuba, Ibaraki, Japan

Molecular hybridization using cloned hepatitis B

virus DNA (HBVDNA) was applied to liver and

tumor tissues from HBV carriers and patients with

hepatocellular carcinoma for determination of state

of HBVDNA in liver and hepatocellular carcinoma.

Free HBVDNA was detected in lliver from thirty-

seven of fifty-two HBV carriers and integration of

HBVDNA into hepatocellular DNA was not iden-

tified in liver from HBV carriers.

Integrated HBVDNA was identified in hepato-

cellular carcinoma tissues from fourteen of fifteen

patients persistently infected with HBV. No inte-

grated HBVDNA was detected in liver tissue ad-

jacent to the tumor. No free HBVDNA was also

identified in liver tissue from a half of patients with

hepatocellular carcinoma.

These results suggest that integration of HBV-

DNA into hepatocellular DNA occurs in conjunc-

tion with malignant transformation.

Hepati t is B virus infection and hepatocellular carcinoma in Taiwan

D.S. CHEN, J .L. SUNG, M.Y. LAI,

J.C. SHEU and H.C. HSU Dept Intern Med, Natl Taiwan Univ

Coll Med, Taipei, Taiwan, ROC

Persistent hepatitis B virus (HBV) infection and


hepatocellular ca rc inoma (HCC) are prevalent in

Taiwan. To study their relationship, the following

studies were performed.

In 247 HCC patients, the serum HBsAg preva-

lence was 88% in contrast to 15% in heal thy adults.

The HBsAg titer tended to be low and did not corre-

late with the frequent ly coexisting m a c r o n u d u l a r

cirrhosis. Ant i -HBc was positive in 98%, but IgM

anti-HBc was present in only 3% of patients. Anti-

HBs was present in 19%. HBeAg/an t i -HBe was

positive in 17% and 79% respectively, correspond-

ing to the infrequest DNA polymerase activities and

serum HBV DNA. In seropositive patients, tissue

HBsAg was present in the non- tumor por t ion in

75-90%, and in HCC tissues in 3 -15%. Tissue

HBcAg was generally absent in these patients, e i ther

in tumor or non- tumor par t .

HBV genome in the liver tissues were analysed

with a sensitive probe and was found in 8 pat ients

studied. It was present in both tumor and non-

tumor tissue. In tegra t ion of HBV genome was found

in all of the 9 tissue specimens conta in ing adequa te

HBV DNA for analysis, including 4 pairs of t umor

and non- tumor liver f rom the same pat ient .

We conclude tha t persistent HBV infection is

common in our HCC patients, and a l though viral

replication is ineffective, the infection is closely re-

lated to HCC in Taiwan.

Close association between activated transforming gene and integrated

H B virus genome in the D N A

from a human hepatoma cell

l i ne (HCC-M)

T. NAKAMURA, T. MORIZANE and

M. TSUCHIYA

Dept Intern Med, Sch Med, Kefo Unfv, Tokyo, Japan

We investigated whether the DNA from a h u m a n

hepa toma cell line, HCC-M, which contains HBV

DNA as in tegrated form had a potent ial of trans-

forming N I H / 3 T 3 into cells with ma l ignan t na ture .

It was demonst ra ted in transfection exper iments

tha t a dose of 50 gg HCC-M DNA was able to induce

about 5 loci of cells growing free from contact in-

hibit ion. It was possible to induce secondary trans-

formants with the DNA from those p r imary trans-

formants. It seemed that those pr imary and secon-

dary t ransformants had a ma l ignan t phenotype

since they grew in semi-solid medium. The integra-

tion of h u m a n DNA sequences, HBV DNA and N-

ras gene in those t ransformants were examined by

the method of Southern using Blur 8, pHBV 114

and N-ras gene as probe. In N I H / 3 T 3 cells N-ras

gene was demonst rab le at two different sites on

agarose gel electrophoresis of Hind III-digested

DNA but the o ther two were not detected. In addi-

tion to the two bands demonst ra ted in N I H / 3 T 3

cells a novel b a n d hybridising to the N-ras probe was

detected only in the t ransformants . One of the

bands hybridising to the HBV DNA was located at

the position of the molecular weight same to the

novel band . These results suggest tha t HB virus

genome and an act ivated N-ras gene of HCC-M are

cotransferred to N I H / 3 T 3 cells and ma l ignan t

phenotypes are provided as the result. T rans fo rming

activity seems to reside in N-ras gene and it seems

likely tha t HB virus genome and N-ras gene are

closely associated on the DNA of HCC-M.

Establishment of a c o n t i n u o u s l y growing cell l i ne (WH257) of woodchuck

hepatocellular carcinoma and its characteristics

K. KOBAYASHI and M. U N O U R A

1st Dept Intern Med, Sch Med, Kanazawa Univ, Ishikawa, Japan

We have established an in vivo nude mouse model

system of woodchuck hepatocel lu lar ca rc inoma

(HCC) (Hepatology 3(5): 633, 1983). This system is

now in the 10th passage, and it produces WHsAg,

confirmed by our made specific monoclonal anti-

body to WHsAg, and the synthesis of a lbumin is also

proven, using an t i - r abb i t ant ibody to woodchuck

a lbumin. So we have tr ied to establish an in vitro

model system of a continuously growing cell line of

woodchuck HCC, using the serially t ransp lan ted

tumor of the nude mouse system.

T u m o r cells derived from the HCC in the 4th

passage were isolated by means of a collagenase per-


fusion technique, and inoculated into plastic dishes.

It has been successfully subcultivated 46 times (8

months) and the doubling time was 26 hours. The

production of WHsAg has not been proven, al-

though the intergration of WHV-DNA in cellular

DNA of WH257 was demonstrated by the Southern

blot technique method, examined by WHV-DNA

probe which was kindly provided by Dr. Gerin in

NIH in the U.S.A.. WH257 produces albumin

proven in its cytoplasm by the indirect immunofluo-

rescent method. This continuously growing cell line

(WH257) provides a definitive model for the study of

oncogenesis of human HCC related to hepatitis B

virus.

Moderators:

Internat ional symposium (3): I m m u n o l o g y and bowel diseases

P.E. H E R M A N S (USA) and Yutaka YOSHIDA (Japan)

Immunological study of lymphocytes isolated from the guinea pig

large intestine

K. OKAWA, K. KOBAYASHI, A. KITANO,

S. KUWAJIMA, H. HASHIMURA,

T. MATSUMOTO, M. HIKI, S. YAMAMOTO*

and Y. KINOSHITA**

*3rd Dept Intern Med and **2ncl Dept

Physiol, Osaka City Univ, Osaka

The purpose of this study was to evaluate the im-

munological characteristics of the large intestinal

lymphocytes in the guinea pig.

Intestinal mucosal lymphocytes (IL) of the large

bowel were isolated by the use of stainless sieve com-

bined with differential centrifugation, and were also

examined lymphocyte responses to polyclonal mito-

gens compaired with mesenteric lymph node lym-

phocytes (MLNL). Guinea pigs were immunized

with ovalbumin (OVA) and Freund's complete adju-

vant, and antigen specific responses of isolated IL

were examined.

Both IL and MLNL responded well to PHA,

ConA and PWM. In contrast, IL did not respond to

LPS, while MLNL responded to LPS. IL isolated

from guinea pig immunized with OVA responded to

OVA.

These results suggested that a majority of isolated

IL was thought to be T-cell, and the presence of the

immunological memory cells was revealed in the

large intestinal mucosa in the guinea pig.

Immunocytochemical observation on solitary lymphoid nodules in human

intestine: Possibity for their participation in local

immune responses

H. NAGURA, Y. SHIODA, Y. TSUTSUMI,

H. HASEGAWA and K. WATANABE

Inst Dis Mechanism ~ Control, Nagoya Univ Sch Med, Nagoya, Dept Surg, Nippon Med

Sch, Tokyo, and Dept Pathol ~ Cell Biol Res Lab, Tokai Univ Sch Med,

Isehara, Japan

Solitary lymphoid nodules are present in the

lamina propria throughout the human intestinal

tract, and look like small Peyer's pathes. If they have

the same functional ability as Peyer's patches, their

role on local immunity must be considerable. In the

present work we have characterized the cells in these

nodules by the peroxidase-labeled antibody method

using monoclonal antibodies detecting T and B

lymphocytes and rabbit antisera to human immuno-

globulins and J chain.

Results: Underlying the covering epithelium were

the lymphoid follicles themselves. The same prin-

ciples for defining T and B areas in the lymphnodes

applied equally well to those nodules. The dome re-

gion contained many macrophages and IgA plasma

cells as well as T and B lymphocytes. T cells in the

areas under the dome as well as in the inter- or peri-

follicular area were mainly composed of Leu 3a-

positive, helper/inducer T cells with a few Leu 2a,


suppressor/cytotoxic T cells. A follicular area con-

tained predominantly small lymphocytes expressing

a B cell surface marker, occasionally with germinal

center, where large and medium lymphocytes of a B

cell lineage and macrophages were present. It is

noteworthy that helper / inducer T cells and Leu 7-

positive, possible NK cells were present also in the

germinal center. This finding seems to be in agree-

ment with the dependence of the germinal center

reaction on the presence of T ceils and NK cells.

A new defect of the transport of IgM into the intestinal lumen in

selective IgA deficiency

T. KANOH*, O. NISHIDA*, H. UCHINO*,

T. MIYAKE** and Y. HISHITANI***

*lst Div, Dept Intern Med, **Dept Geriat, Kyoto Univ, Kyoto, and ***lst Div, Dept Intern Med, Osaka Med Coll,

Takatsuki, Japan

In selective IgA deficiency a large number of IgM- cells appear in the GI mucosa instead of IgA-cells

and IgM was detected on the luminal surface as a secretory form binding SC. In a 49-year-old woman with Sjoegren's syndrome (SS) having selective IgA deficiency, no IgM was found on the surface, al-

though there were many J chain positive (J+) IgM- cells in the lamina propria. In vitro binding of SC to IgM on the colonic sections was investigated in this case and another case of selective IgA deficiency (S.Y.) as a control whose colonic mucosa showed the localization of IgM on the lumina surface. The former showed no binding of free SC to the cytoplasm of IgM-cells. Then, in order to study the capability of IgM from this case in binding SC, the following was performed. Using the antiserum against inaccessible antigens od SC, we determined the remainder of free SC after the incubation of polymeric Ig (pig). The more the remainder, the

less the capability of p lg in binding SC. Two monoclonal IgM (J+) and four monoclonal pIgA (J+) were prepared. The % of SC-linked IgM or pIgA was 49% to 56% or 34% to 100%, respectively. Cohn Fr II and human albumin could not bind free SC at all. Only 3% of IgM from the patient bound free SC, whereas 42% of IgM from the case S.Y.

These interesting findings shown herein indicate a new defect of p ig transport system in the GI mucosa, which seemed to depend upon molecular defect of the patient's lgM. This is also one of abnormalities in IgM system in SS.

Immune response in intestinal

tuberculosis

D.K. BHARGAVA, B.N. TANDON, SHRINIWAS,

B.M.L. KAPUR, T.C. C H A W L A and

Usha KIRAN

All India lnst Med Sci, New Delhi, India

The immune response is one of the most potent

host factor against infection and its study could be

pertinent for morphological expression of disease

and demonstration of antibodies may help in evolv-

ing a serological test.

30 patients having ileal, ileocecal and colonic

tuberculosis were included in the study. The diagno-

sis was confirmed by histology and /o r bacterial

isolation. The immune response was evaluated by

PPD test, DNCB (Dinitrochlorbenzene) skin sen-

sitization, leukocyte migration inhibition (LMI) and

soluble antigen fluorescent antibody test (SAFA).

The PPD response was significantly positive in

95.45% patients. DNCB was negative in 45% and

positive in 55% (majority 2+) of patients. LMI was

positive in 77%. There was good correlation be-

tween PPD response and LMI. These findings indi-

cate that there is a subtle defect in cell mediated im-

mune response in patients of intestinal tuberculosis.

The SAFA test was positive in 25 (83.3%) of 30 pa-

tients. The false positive result was observed only in

one of the 56 control subjects. Thus SAFA test is

more specifically related to existance of active intes-

tinal tuberculosis and differentiates it from other

non-tuberculosis intestinal diseases.

Diagnosis of intestinal tuberculosis using enzyme immunoassay

J .p. GUPTA, A.K. JAIN, B.K. A G R A W A L

and Saroj GUPTA

Div Gastroenterol, Inst Med Sci, Banaras Hindu Univ, Varanasi-5, India


The diagnosis of intestinal tuberculosis often pre-

sents difficulties and many a t ime requires laparo-

tomy. As such, the usefulness of a sensitive serologi-

cal test in its diagnosis needs evaluation.

In the present study sera from 38 histologically

established cases of intestinal tuberculosis, 26 of pul-

monary tuberculosis (+ve control) and 20 healthy

subjects ( -ve control) were studied for the titres of

antibodies to PPD employing ELISA. The titres

were expressed in terms of optical density (O.D.).

Mean titre in intestinal tuberculosis was 0.282

(range 0.114 to 0.532), in +ve controls 0.536 (range

0.074 to 0.98) and in -ve controls 0.121 (range

0.062 to 0.149).

The diagnostic usefulness of the test (sensitivity)

was evaluated taking 100% specificity i.e. highest

value among the negative controls (0.149) as refer-

ence point. 84% of intestinal tuberculosis and 88%

+ve controls had diagnostic titres of 0.150 or more.

These findings suggest usefulness of antibody titre

to PPD in the diagnosis of tuberculosis. However,

further confirmation and its relationship with ac-

tivity of the disease remains to be established.

NK activities and N K (HNK-1 +) cells in familial polyposis coli

M. CHIBA

1st Dept Intern Med, Akita Univ Sch Med

Natural killer (NK) activities and NK cells were

studied in 14 patients with familial polyposis coli

(FPC). Peripheral blood lymphocytes (PBL) from

FPC showed normal cytotoxicities against K562,

P4788 derived from colon cancer, and Chang cells.

Lamina proprial lymphocytes (LPL) isolated from

the resected colons exhibited expected cytotoxicities

against K562 and Chang cells. NK cells identified by

HNK-1 monoclonal antibody were distributed

mostly in a normal range for PBL and were not con-

tradictary to those seen in the controls for LPL. As

these results were not modified by the time factor of

colectomy, it seems unlikely that the deficit or im-

pairment of NK activities is one of the backgrounds

for the development of adenomas and the malignant change in FPC.

Interferon production activity of regional lymphode lymphocytes in patients

with colo-rectal cancer

M. SASAKI, M. KONN, Y. YAMANAKA,

T. MORITA, T. HASHIZUME, K. NARA,

H. ODAGIRI , M. FUJII and

K. ONO

Dept Surg, Hirosaki Univ Sch Med,

Hirosaki, Japan

Accumulative evidences have demonstrated the

anti- tumor effects of interferon (IFN). This study

was undertaken to investigate the roles of regional

lymphnode lymphocytes (RLL) in host defence

against tumors by testing the IFN production

ability.

Materials and Methods: Mononuclear cells were

isolated from regional lymphnodes and peripheral

blood (PBL) by Ficoll-Isopaque density gradient

centrifugation. IFN was induced by in vitro culture

with PHA-P, OK-432 and various cultured cell

lines. Determination of IFN activity were performed

by the cytopatbic effect reduction in VSV chal-

lenged human WISH cells.

Results: Results are summarized as follows:

1) No correlation was observed between the IFN

production ability and clinical stages, and IFN was

still detected even in cases of advanced stages.

2) Two types were demonstrated in IFN produc-

tion pattern according to the difference of IFN in-

ducers. IFN induced by OK-432 showed almost the

same level in both RLL and PBL.

3) IFN level induced by PHA-P was significantly

higher in RLL as compared with PBL. On the con-

trary, IFN induced by K562, HL-1 and HL-2 (lym-

phoblastoid cell lines) were significantly higher in

PBL than in RLL. This might be due to the fact

that NK cells are quite few in regional lymphnodes.

Thus, it was suggested that the function of RLL and

PBL was different in immune responses.

Immunological defense of gut-associated lymphoid tissue in gastrointestinal

malignancies

Y. NIO, T. TSUCHITANI , T. INAMOTO,

H. KODAMA and T. TOBE

282 Proceedings of26thAnnuaIMeeting Vol. 20, No. 3

Dept Surg, Univ of Kyoto Med Sch, Kyoto, Japan

We studied the immune response of gut-as-

sociated lymphoid tissue (GALT), especially mesen-

teric lymph node (MLN) and Peyer's patch (PP), in

murine experimental cecal tumors, which were pre-

pared by the transplantation of tumor cells into the

cecal patch, using syngeneic tumor-host system

(BALB/c mouse-BAMC 1 fibrosarcoma, and DS

mouse-SC 42 mammary adenocarcinoma).

As immunological parameters, Natural Killer

(NK) activity, lymphoblastic response to PHA

(LBR) and mixed lymphocyte tumor reaction

(MLTR) in mice with these tumors were compared

with those in sham-operated mice.

In spleen cells, LBR was remarkably suppressed

from one week after transplantation, but NK ac-

tivity and M L T R increased between one and two

weeks, and then decreased three weeks after trans-

plantation. In MLN cells, moderate suppression of

LBR and slight decrease of NK activity were ob-

served one week after. M L T R was enhanced be-

tween one and two weeks, but decreased thereafter.

In PP cells, these parameters were only slightly influ-

enced by tumors.

To augment the ant i- tumor immunity of G A L T

in gastrointestinal malignancies, we administered

the sterptococcal preparat ion "OK-432" orally. In

mice with transplanted cecal tumor, LBR, NK ac-

tivity and M L T R were augmented in MLN cells, but

not in spleen cells, by oral administration of OK-

432. In PP cells only NK activity was increased. In

addition, tumor growth was suppressed. These re-

sults indicate oral administration of OK-432 aug-

ments the local anti- tumor immunity of G A L T and

may have favorable therapeutic effects in cancer

therapy.

Monoclonal ant ibodies derived human colorectaI cell l ine by hybr idoma

techn ique (pre l iminary report)

Z.R. SHI and Y.S. KIM*

Dept Med, 2nd A)~liated Hosp, Hunan Med Coll, Changsha, Hunan, China and *GI Res Lab, VA Med Cent and Dept Med,

Univ of California, San Francisco, CA 94121 USA

Two monoclonal antibodies, LsL14 and LsL123

derived by fusion of mouse myeloma cells with

spleen cells from mice immunized with a human

colorectal carcinoma cell line, Ls 174T is described.

Both LsL14 and LsL123 were identified as mouse

IgM (u, k). These antibodies, when reacted with 10

different cultured human cancer cell lines, includ-

ing colonic cancer, gastric cancer, melanoma, lung

cancer, pancreatic cancer, bladder cancer and a

human fibroblast cell line using enzyme immunoas-

say technique, showed that LsL123 only reacted to

Ls174T, and LsL14 reacted to both Ls 174T and a

gastric cancer cell line Hutu 80. Immunohisto-

chemical study showed that LsL123 and LsL14 posi-

tively reacted to colorectal adenocarcinoma tissue in

44.5 % and 55.6 % respectively by using indirect im-

munofluorescence technique. But they also cross

reacted to normal human colonic tissue on luminal

membrane of lower colonic crypt (60 % and 30 %) or

on goblet cell mucin (30% for LsL14). This result

indicated that these two monoclonal antibodies were

not specific to the surface antigen of human colorec-

tal adenocarcinoma cells.

Immunohistological studies in various bowel diseases using monoclonal

antibodies

H. YAMAOKA, K. NISHIYAMA, S. TAJIMA,

T. FUKUSHIMA and S. TSUCHIYA

2nd Dept Surg, Yokohama City Univ, Yokohama, Japan

Distribution of T lymphocyte subsets in various

bowel diseases was analysed in tissue sections using

monoclonal antibodies. Ulcerative colitis, Crohn's

disease, familial polyposis coli, adenomatous polyp,

colorectal carcinoma and non-involved colon during

colectomy for carcinoma were studied. Indirect en-

zyme immunoassay was performed. OKT3, OKT4,

OKT8 (Ortho Labs.; 1 in 25), Leu M2, Leu 10 and

HLA-DR (Becton-Dickinson; 1 in 25) were used as

the first layer, and goat IgG F (ab')2 anti-mouse

(IgG + IgM) labelled with horseradish peroxidase

(Tago; 1 in 25) was used as the second layer. In


ulcerative colitis, OKT4 + cells were predominantly

infiltrated into the mucous membrane. In Crohn's

disease, OKT8 + cells were dominant on the con-

trary. OKT4 + cells were dominant in familial poly-

posis coli, and in adenomatous polyp. OKT3 § cells

were abundant and OKT4+ cells were dominant

around colorectal cartinoma. The proportion of

OKT3 + cells was significantly increased in ulcera-

tive colitis and colorectal carcinoma respectively

(p<0.01) Likewise, that of OKT4 + cells was in-

creased in ulcerative colitis (p<0.05) and well differ-

entiated adenocarcinoma of colo-rectum (p<0.01).

In Crohn's disease, the proportion of OKT4 + cells

decreased (p<0.05) and OKT8 + cells increased

(p<0.01).

Significant difference was observed in the dis-

tribution of intestinal lymphocyte subpopulation of

the various diseases, especially in ulcerative colitis

and Crohn's disease.

Studies on colon antigens by monoclona l

an t i bod i e s - -The re la t ionship of

ulcerat ive colitis with

colon ant igens

K. KUROE, A. KIMURA and Y. SUWA

1st Dept Intern Med, Hirosaki Univ Sch Med, Hirosaki, Japan

Histopathological analysis of ulcerative colitis

(UC) tissue has shown numerous inflammatory

mucosal infiltrates consisting of lymphocytes and

plasma cells in the involved colon. These mononu-

clear cells might be responsible for complicated cell

interactions, autoimmune reactions to a ceratin

antigen, and eventually damaging colon epithelium

cells. Helper and suppressor T cells have important

roles in the regulation of immunologic responses,

and their imbalance produces autoimmune phe-

nomena. We have shown that con A induced sup-

pressor activities of colonic mucosal lymphocytes

and PBL in UC were significantly lower than those

in normal controls. It was then suggested that a spe-

cific colon antigen induced the autoimmune reac-

tions with the suppressor activity decreased. There-

fore, the colonic antigen which is capable of in-

ducing anti colon antibody and sensitized lympho-

cytes was analysed by a hybridoma technique. Two

kinds of monoclonal antibodies for human colon

antigens were produced. One was against the base-

ment membrane and a part of Iamina propria of

healthy colonic mucosa. It reacted specifically with

the colonic mucosa. The other was against secretory

granules in the goblet cells of healthy large and

small intestine and against mucin. A monoclonal

antibody for rat colon antigen was produced, and it

reacted specifically with the goblet cells of the rat

colon. The antigens were detected by these mono-

clonal antibodies. From the above results followings

were speculated. Antigen of the basement mem-

brane or goblet cells may induce the autoimmune

reactions; Colonic cells may be damaged by the sen-

sitized lymphocytes and /o r ADCC, or by an altered

local defence mechanism of mucus. The above

speculation is supported by the findings reported

that the lymphocytes are cytotoxic for colonic cells

and that the goblet cells are decreased in active stage

and increased in remission in UC.

Significance of mucus deplet ion of colon

goblet cells in u lcera t ive colitis

H. FURUKAWA* and T. MIYAKE**

*Dept Med, Otsu Red Cross Hosp, Otsu, **Dept Geriat, Univ of Kyoto,

Kyoto, Japan

The phenol-water extract of human fetal colon

(Broberger et al. 1959) was used by us as an antigen

of skin test in patients with ulcerative colitis (u.c.)

and showed high frequency in the delayed reaction.

The extract also produced delayed reaction in

guinea pigs experiments and the immune sera con-

tained precipitating antibody. Immunofluorescence

method with the antisera against the polysaccharide

fraction of the extract using human colon specimens

showed specific fluorescence in cytoplasm of goblet

cells. Therefore, autoantibodies of u.c. are against

the goblet cell mucus. Using the anti-mucus, anti-

SC and anti-IgA fluorescent antisera, we examined

the biopsy specimens of the patients. In u.c., even in

the normal parts, SC and IgA in lumen of the glands

were diminished. Moreover, in the diseased parts,

mucus depletion was noted. The border parts decide

284 Proceedings of 26thAnnualMeeting Vol. 20, No. 3

the fate of the disease. Mucus depletion is integral

characteristics of u.c. Mucus glycoproteins are one

of the most important defence factors and their de-

pletion lays the epithelia bare, thus realizing the in-

vasion of bacteria, lectin-like agents and food aller-

gens. It is not conclusive whether autoimmunity in

u.c. is primary or secondary. However, it plays one

of the important roles in mucus depletion.

Abnormalities of immunoregulatory T-cel l popula t ions in patients

wi th ulcera t ive colitis

M. WATANABE, T. YOSHIDA, S. AISO,

H. ASAKURA and M. TSUCHIYA

Dept Intern Mecl, Sch Med, Keio Univ, Tokyo, Japan

Circulatory T-cells were characterized in 42 pa-

tients with ulcerative colitis (UC) by a fluorescence

activated cell sorter (FACS), using monoclonal anti-

bodies to the cell surface antigens of suppres-

sor/cytotoxic T-cells (Anti-Leu-2a) and helper/ in-

ducer T-cells (Anti-Leu-3a). The population of

Leu-2a positive cells was decreased in patients with

active UC, while that of Leu-3a positive cells did not

show any remarkable change. Moreover, function of

circulating suppressor cell was revealed to be im-

paired in the patients by Con A-induced suppressor

T-cell function assay.

Also, we studied the cell surface characteristics of

thymus cells from UC patients who had undergone

thymectomy for association of thymic hyperplasia.

Although the control thymus consisted of 82-94 % of

cells reactive with Anti-Leu-2a and 90-95% of ceils

reactive with Anti-Leu-3a, both Leu-2a positive and

Leu-3a positive cells were decreased.

Next, the relationship between lymphocytophilic

autoantibodies and peripheral T-cell subsets was ex-

amined by cell sorting and double staining tech-

nique on a FACS. The lymphocytophilic antibodies

in patients with UC were demonstrated to be mainly

reactive with Lau-2a positive cells and also, they

were reactive with some populations of Leu-3a posi-

tive cells and non-helper, non-suppressor T cells.

The other factors that may influence the immuno-

regulatory system in UC patients will be also dis-

cussed.

Cytokine product ion capacity in

inflammatory bowel disease

M. MIURA and N. H I W A T A S H I

3rd Dept Intern Med, Tohoku Univ Sch Med, Sendai

Immunological impairment in inflammatory

bowel disease has been the title of a number of

papers. In the present study we investigated presence

or absence of impairment in cytokine production

from mononuclear cells in inflammatory bowel dis-

ease. In addition T cell subsets and Ia positive

monocytes were examined. The study subjects con-

sisted of 26 cases of Crohn's disease (CD) and 7 cases

of ulcerative colitis (UC). As the controls, healthy

donors (HD) matching age and sex were used. No

differences were seen between UC and HD in any of

the parameters examined. However, reduction in

interleukin 2 and interferon activities was noted in

CD. They can not be attributable to decreased num-

ber of T cells, because no significant differences

were seen in the number of T cells including subsets.

On the other hand, as to the interleukin 1 produc-

tion capacity no differences were seen between HD

and CD. These results suggest that some form of dys-

functions exist at the level of T cells in the peri-

pheral blood in CD.

A novel IgA protease f rom Clostridium sp. (M.O.-6), an isolate f rom fecal

flora of the pat ient with I .B .D. ,

capable of cleaving IgA1 and IgA2:A2m (1), but not

IgA2:A2m (2) paraproteins

Y. FUJIYAMA, K. KOBAYASHI, T. BAMBA,

S. HOSODA and Y. BENNO

Dept Med, Shiga Univ Med Sci, Otsu, Dept Pediatr, Sch Med, Univ of

Yamaguchi, Ube, and Inst Physical and Chem Res,

Wako, Japan

In the course of our study examining the produci-


hility of IgA1 protease in the members of intestinal flora from patients with I.B.D.s to evaluate it's viruluency in the pathogenesis of I.B.D., we found out a unique strain of genus Clostridium which pro-

duced endopeptidase capable of cleaving both human IgA1 and IgA2 paraproteins.

The treatment of both IgA1 and IgA2:A2m (1),

myeloma proteins with the culture filtrates of

Clostridiurn sp. (M. O.-6) resulted fragmentation of these proteins into 2 fragments compatible to Fab and Fc on immunoelectrophoretic analysis. The MW of Fc fragment of IgA1 determined by SDS polyacrylamide gel electrophoresis was larger than

the Fc fragments obtained by the treatment with IgA1 proteases from previously reported bacterial strains. N-terminal aminoacids of Fc fragments obtained from both IgA1 and IgA2:A2m (1)were consisted of Valine. IgA2:A2m (2) paraproteins and also IgG, albumin, bovine serum albumin showed resistence to this endopeptidase.

These data are suggesting that the cleavage site of this novel endopeptidase from Clostridium sp. (M.O.-6) is within the homologous aminoacid se-

quence (221-222) lying in the hinge region of both IgA1 and IgA2:A2m (1) heavy chains.

Experimental inflammatory bowel disease in mice

Y. WAKASHIN, M. WAKASHIN, T. MORI, S. UEDA, H. YOSHIDA, Y. MORI

and K. OKUDA

Dept Med, Univ of Chiba, Chiba, Japan

It has been suggested that immunological mecha- nisms are involved in the etiology of inflammatory bowel disease (IBD), but the evidence is inadequate. In this study, we aimed to produce an experimental IBD model in mice, by injection of anti-colon antibody, rabbit IgG sensitized T cells, and/or colon antigen sensitized T cells, and the relation of pathological lesions to cellular immunity was determined.

In group 1, 70 BALB/c mice were injected with anti-colon rabbit IgG and rabbit IgG sensitized T

ceils, and in group 2, 20 BALB/c were injected with syngeneic colon antigen sensitized T cells. Colon

antigen was prepared from BALB/c mouse colon mucosa, after washing with saline, homogenization and trypsinization. Antiserum was prepared by im- munizing rabbits with 100 mg of colon antigen 3 times. T cells were obtained from lymphnodes of BALB/c mice sensitized with rabbit IgG, and colon antigen. Histology and circulating antibody were examined at various time intervals. In group 1, IBD occurred with a high incidence in 70% of examined

mice, and the changes included focal necrosis, infiltration of mononuclear and polymorphonuclear cells in the lamina propria. And in group 2, changes

were noted in 60% of mice, and it was characterized by typical ulceration on one side of the colon with chronic inflammatory cells infiltration in the ulcer base. These pathological changes seem to be closely related to cellular immune reactions, especially colon antigen sensitized T cells.

Internat ional Free Paper Session (1): Moderators: N. M A D A N A G O P A L A N (India) and Tatsuro T A K I N O (Japan)

A study of antibody to liver-specific membrane lipoprotein (anti-LSP) in circulating immune complexes

of patients with liver diseases

Li YOU and Ye WEI-FA

Bethune Med Coll, Changchun, Jilin, China

To study the compositions of liver-specific anti-

gen-antibody in circulating immune complexes, we

have measured anti-LSP in circulating immune complexes in 62 patients with liver diseases with positive immune complexes (ICs) in their sera, com- bining solidphase radioimmunoassay to anti-LSP with PEG precipitation method and glucine buffer separation method. To confirm the specific results, we have established competitive inhibition group be- sides standard control group during measurement. 5 of 8 patients with acute hepatitis are ICs-anti-LSP positive, 23 of 26 with chronic hepatitis and 20 of 28


with cirrhosis are positive. There is no statistic

difference between HBsAg (+) group and HBsAg

( - ) group. Our result suggests tha t there is LSP-

anti-LSP complex in ICs of patients with liver dis-

eases.

The function of pituitary adrenal axis in hepatic cirrhosis

Guang-Xao YANG and Wei-Fa YE

Dept Med, Bethune Med Coil, Changchun, Jilin, People's Republ of China

To study the funct ion of pi tui tary adrenal axis, we

observed the rhy thmic secretion of adrenocort ico-

sterol day and night , and the response of p lasma

corticosterol to external ACTH and dexamethason

in patients with decompensated hepatic cirrhosis

(n=21). The rhy thmic secretion of corticosterol was

present, but the level of plasma corticosterol at 8:00

a .m. was lower in the cirrhosis group than that in

the normal control group (8.64 + 4.21 vs 14.9 •

4.96 g g % , p<0.01) . The response peak of p lasma

corticosterol appeared 8 hours after ACTH adminis-

t ra t ion in the control group, whereas it was not pre-

sent unti l after 16 hours after ACTH adminis t ra t ion

in the cirrhosis group. Th e response peak values to

ACTH was also reduced in the cirrhosis group

(21.02 • 8.41 vs 35.84 • 9 . 7 4 / / g % , p<0.05) . Four

of the ten pat ients examined, desamethason escaped

the inhibi tory effect of dexamethason on cortico-

sterol secretion. The da ta suggest tha t funct ional

disorder of pi tui tary adrena l axis be present in pa-

tients with cirrhosis. We conclude that it results

from decreased funct ion of adrenocortex and defect

of hormonal degrada t ion in patients with cirrhosis.

Local aspects of hepatic infections and impaired liver function

R.H.H. N E L W A N

Div Tropical and Infec Dis, Dept Med, Fac Med, Univ of lndonesia Dr.

cipto Mangunkusumo Hosp, Jakarta, Indonesia

As developing countries are struggling to improve

their s tandard of living, serious problems are some-

times encountered in heal th care. One of the prob-

lems that has to be faced with in Indonesia is the

high risk of liver infections and impai red liver func-

tion.

Liver impa i rmen t on the other h a n d calls for

more proper drug prescript ion habits. Many drugs

may affect the liver in a harmful way if not used

judiciously.

The wide spread use of local herbal medicine tha t

also may conta in hepatotoxic substances seems also

to contr ibute to the impa i rmen t of the normal liver

funct ion in our populat ion.

An accepted belief tha t only hepati t is B infections

are associated with hospital cross infections was

proven not to be the case. A case will be presented

on the occurrence of hepati t is A infection in a hospi-

talized pat ient and the hypothetical mode of trans-

mission.

Cell Mediated Immunity in amoebic l ive r abscess

B.M. GANDHI , H. GUPTA, M. IRSHAD,

T.C. C H A W L A and B.N. T A N D O N

Dept Gastroenterol, AIIMS, N. Delhi, India

Pathogenesis of invasive amoebiasis remains un-

known. Present study was under taken to study the

cellular i m m u n e response by leucocyte migra t ion in-

hibi t ion and count ing of pr imary effector cells i.e.

thymus derived (T) cells and bone mar row derived

(B) cells in 26 pat ients of amoebic liver abscess

(ALA) and compared with tha t of non-invasive in-

fections viz. 11 cyst passers, in 2 pat ients of acute

amoebic dysentry and in 3 pat ients of hepat ic

amoebiasis. 20 heal thy controls were also included

in the study. The study in ALA was repeated after

t rea tment and clinical recovery. A positive response

of LMI was ob ta ined in 16/26 acute ALA, whereas

9 /20 after recovery. No response was found in other

groups.

Results of the present study show a t ransient state

of immunodepress ion in patients of ALA as ob-

served by significant decrease in n u m b e r of T-active

(P<.001) and T- to ta l cells (P<.01) in ALA before

and after the recovery, however no significant differ-


ence was found in other groups. It thereby may help

E, histolytica to evade the defence mechanism of the

host to invade target cells and produce liver abscess.

The immune response was reversed after treatment

and clinical recovery in ALA.

Tc-99m DISIDA scintigraphic studies in the diagnosis of hepatobiliary

diseases

Mong CHO and Bang-Hyun LIU

Dept Intern Med, Coll Med, Pusan Natl

Univ, Pusan, Korea

For evaluation of the diagnostic value and useful-

ness of the Tc-99m DISIDA in the hepatobiliary dis-

eases, the hepatobiliary scintigraphy was performed

in 76 patients and 25 normal subjects at Pusan

National University Hospital during the period from

March 1983 to January 1984.

In ten cases of chronic cholecystitis, they showed

delayed visualization of the GB in two cases, and de-

layed biliary to bowel transit times in other two

cases.

In nine cases of partial biliary obstruction, they

showed none of the hepatobiliary elements but he-

patic uptake in two cases, and delayed visualization

of duodenum till 120 minutes in one case but nor-

mal scintigraphic finding in others.

In ten cases of hepatoma, the visualization of

hepatobiliary elements were dependent on size, loca-

tion and involvement of the regional lymph nodes.

The common hepatic duct, common bile duct,

GB, and duodenum were clearly visible in the cases

of serum bilirubin level showing less than 15 mg/d l ,

and also even visible in the case of chronic active

hepatitis showing 25 mg /d l serum bilirubin.

Conclusively Tc-99m DISIDA scintigraphy was

helpful in the diagnosis of cholecystitis, partial bili-

ary obstruction, and involvement of hepatobiliary

system of hepatoma.

Diagnostic value of fasting and postprandial serum conjugated cholic

acid in chronic liver disease

Xie-ning WU Div Gastroenterol, Dept Med, Munic

1st People's Hosp, Shanghai, China

Fasting and postprandial serum conjugated cholic

acid (CCA) was studied by radioimmunoassay in 163

patients of various liver diseases and 97 healthy sub-

jects. Fatty meals of 55 and 88 gin gave similar re-

sults (p>0.05), the former was more tolerable in

Chinese. The mean fasting and postprandial CCA in

various diseases in increasing order were: CPH,

schistosomal cirrhosis, CAH, posthepatitic cirrhosis

and CAH cholestatic type. CPH vs healthy subjects

both were p<0.01; CPH vs CAH both p<0.001;

CPH vs schistosomal cirrhosis, fasting CCA p<0.01,

postprandial CCA p<0.001; posthepatitic cirrhosis

vs schistosomal cirrhosis fasting level p<0.01-0.05;

posthepatitic cirrhosis vs CAH both p<0.05-0.1.

Both fasting and postprandical CCA were 100%

positive in CAH and 93.3% and 85% respectively in

posthepatitic cirrhosis, much higher than other rou-

tine liver tests, whereas in CPH, they were not so.

Elevation of fasting CCA level could be categorized

into four grades: mild, moderate, conspicuous and

very conspicuous. Linear correlation was seen be-

tween fasting and postprandial CCA, but not with

other liver function tests. In conclusion: the sensi-

tivity of fasting and postprandial CCA varies with

different liver diseases, it is most sensitive and valu-

able in diagnosis of CAH and posthepatitic cirrho-

sis, but it can't replace the conventional liver tests in

CPH. To the authors' opinion, postprandial CCA is

not the most sensitive liver function test as advocated

by others.

Gallstones of intermediate cholesterol content in Northern India

Rakesh K. T A N D O N , A.K. SRIVASTAVA,

B.M.L. KAPUR and R.K. MISRA

Dept Gastroenterol, Surg ~ Biophysics,

All India Inst Med Sci, New Delhi, India

Of 152 consecutive gallstone patients operated at

our centre in Northern India, 122 (80.2%) were

found to have cholesterol gallstones (CS) and 10

(6.6%) pigment stones (PS), their individual choles-

terol contents being 70% and <20% of dry weight,

respectively. The remaining 20 (13.2%) patients


had stones with cholesterol contents in between 20

and 70% of dry weight and were therefore referred

to as gallstones of intermediate cholesterol content

(IS). Gallstones from 46 patients and bile samples

from 38 of them were analysed. The mean choles-

terol content of the stones (% dry weight) and the

cholesterol saturation index (CSI)of bile were 85.10

-+ 8.67 and 1.67 -+ 0.69 for CS, 6.07 + 3.57 and 0.83

_+ 0.35 for PS, and 48.10 _+ 17.92 and 2.54 _+ 1.24

for IS respectively. The CSI of control bile samples

was 0.97 _+ 0.30. Thus, CS as well as IS were asso-

ciated with bile supersaturated with cholesterol.

The lower cholesterol content of IS as compared

with CS was made up by a significantly higher con-

tent in IS of calcium, bilirubin, protein and muco-

polysaccharides. Samples of 5 IS were further

studied by X-ray diffraction, infra-red spectroscopy

and scanning electron microscopy. The findings sug-

gested that IS were cholesterol stones with an excess

deposition in them of non-cholesterol substances.

The large amount and the type of distribution of

calcium and pigment compounds in IS might be an

impediment to their dissolution by cholelitholytic

agents.

In ternat iona l Free Paper Session (2):

Modera to r s : B a n g - H y u n L I U (Korea) and Yoshio T A O K A (Japan)

Medical problems and their management in liver transplantation

Shi-huJIANG and Yan-zun LIN

Dept Med, Rui-jin Hosp, Shanghai 2nd Med Coll, Shanghai, China

Homologous orthotopic liver transplantation was

performed in six cases of hepatoma. Four medical

aspects concerned were: 1) selection and prepara-

tion of the patient. Internist should assure the tumor

confined only to the liver and a good coagulation

mechanism before surgery. 2) management of 'ahe-

patic phase'. Hemorrhagic diathesis due to lack of

factors VII, IX and X, increase of fibrinolysis and

heparin-like substances should be corrected by

PPSB, trasylol and protamine respectively. Meta-

bolic acidosis and hyperglycemia were frequent and

must be corrected. 3) management of graft rejec-

tion. Acute rejections occurred regularly on 4th-7th

day and 10th-15th day after operation, but irregu-

larly in subsequent attacks. There were a total of 32

such episodes, each time was well controlled with

bolus injection of methyl prednisolone. We found

sudden decrease of bile excretion a premonitory sign

of impending rejection. 4) management of compli-

cations of immunosuppression. Upper GI bleeding

occurred in first two cases but not in subsequent four

after giving prophylactic cimetidine. Opportunistic

bacterial infection could be controlled by antibio-

tics, but one case contracted acute fulminating HBV

hepatitis. All six recovered from surgery but sur-

vived 54 261 days only, averaged 156.7 days. Causes

of death were bleeding in one, acute hepatic failure

in one and recurrence of hepatoma in four. Our ex-

periences indicate patients with hepatoma are not

good candidates for liver transplantation.

Clinical and scintigraphic studies on id iopathic por ta l hypertension

in Korea

Bang-Hyun LIU

Dept Intern Med, Coll Mecl, Pusan Natl Univ, Pusan, Korea

Clinical & scintigraphic studies on 54 cases of IPH

were carried out at the Pusan University Hospital

between the period of 1962-1982.

They involved 29 males and 25 females.

Male:female ratio was 1.2:l . Majority cases

(63%) were found in younger age (between age

20-39).

Hematologically anemia, leukopenia and throm-

bocytopenia were significant but chemical liver

function abnormality was minimum.

On immunological studies he found HB Virus

markers in 4 cases out of 20 patients.

In this study the author found huge spleno-

megaly, portal hypertension and pancytopenia. His-


topathologically congestive changes and Gandy-

Gamma nodule (Fibroadenie) was also characteris-

tic in splenectomized spleen but hepatic dysfunction

was slight and minimal fibrosis was noticed on liver

biopsy. And on radiogold scintigram marked spleen

visualization was characteristic in all cases, however

no significant changes of hepatic figure and uptake.

Correlation between high portal pressure and

spleen visualization was significant and correlation

between spleen weight and spleen visualization also

noticed.

Sonography of portal venous system in

portal hypertension: Correlation with the size of oesophageal

varices

S.A. ABDURACHMAN and Sujono HADI

Div Gastroenterol, Dept Intern Med, Padjadjaran Univ Sch Med /Hasan

Sadikin Hosp, Bandung, Indonesia

Portal venous system was studied with sonography

in patients with endoscopically documented variceal

bleeding. Optimal scans were obtained in 60 pa-

tients.

Dilatation of the portal venous system may occur

in portal hypertension, and the diameter of portal,

splenic and superior mesenteric veins greater than

13 rnm, 10 mm and 8 mm respectively, were indica-

tive of portal hypertension with a sensitivity of 60 %,

74% and 55% respectively.

There was no significant relationship between the

variceal size and the diameter of portal, splenic and

superior mesenteric veins, although larger varices

seem to be associated with a wider portal venous sys-

tem.

It is concluded that in patients with UGI bleed-

ing, UGI endoscopy should be followed by sono-

graphy of the portal system. Variceal bleeding will

be demonstrated by endoscopy, while sonography is

a valuable screening procedure in the evaluation of

portal hypertension and provides valuable informa-

tion regarding the caliber and patency of the portal

venous system.

Endoscopic sclerotherapy (EST) in the

long term: Management of portal

hypertension

D.K. BHARGAVA

Dept Gastroenterol, All India Inst Med Sci, New Delhi, India

The repeated EST procedures at intervals has

been undertaken to evaluate its value in prevention

of recurrent bleeding and eradication of varices.

82 patients with an underlying cirrhosis in 51 and

noncirrhotic diseases in 31 were included in the

study. Polidocanol 1% was injected intravariceally

at an interval of 3 weeks till the varices were obliter-

ated. The patients were followed from 2 to 34 mth

with an average duration prior to EST was 11.43

and post EST follow up was of 10.88 mths. Alto-

gether prior to EST there were 204 episodes of

bleeding & 681 units of blood were transfused. In

contrast in post EST period they have 48 episodes of

bleeding and 70 units of blood was needed. Recur-

rent bleeding after EST was more in cirrhotic than

non cirrhotic group and majority of them were seen

prior to 6 EST injections. The varices disappeared

in 40 (74%) out of 54 patients. 28 patients are still

early for analysis. None of them bled after eradica-

tion of varices. In 2, varices reappeared after 12 and

16 mths. 6 (7.3%) patients had complications as

stricture and ulcer. 14 (28%) patients died among

50 patients classified as child's B and C. 10 of them

were cirrhotic. It is difficult to comment on quanti-

tative survival. Thus EST has decreased recurrent

bleeding and need of blood transfusion thereby

quality of life improved. Varices can be obliterated

in majority of patients.

Modified injection sclerotherapy of esophageal varices

B.K. A G R A W A L and K.R. BHAT

Dept Med, Kasturba Hosp, Manipal, India

A prospective randomised study comparing the

efficacy of intravariceal injection sclerotherapy with

or without propranolol administration was done in

27 consecutive pts. (20 cirrhosis, 2 NCPF, 6 portal

vein obstruction) in whom initial active variceal

290 Proceedings of 26th Annual Meeting Vol. 20, No. 3

bleeding was controlled by conservative methods.

Oral propranolol was given in the dose to reduce the

resting pulse rate by 25%. The two groups (14 with

& 13 without propranolol) did not differ in age, sex,

cause of portal hypertension, Child's grade or grade

of varices significantly. Intravariceal sclerotherapy

was done with Olympus GIF-Qscope and flexible in-

jection cannula using 1% Aethoxysklerol. Injections

were repeated at interval of 1 to 3 weeks till the

varices were obliterated.

The recurrence of bleeding within first 24 hours

was significantly low in the propranolol treated

group than in the control group. The number of

courses of injections required to obliterate the

varices in the propranolol treated group was also sig-

nificantly less than that in the control group.

Concl.: Intravariceal sclerotherapy in patients

pretreated with propranolol leads to early oblitera-

tion of varices & also reduces risk of early rebleed-

ing.

method, the mortality was 33%. After 1977, herbs

have been used in combination with the conven-

tional western treatment. A Sengstaken tube is

passed routinely for temponade. Herbal mixture

compounded of Rheum and Bletilla striata is intro-

duced into the stomach through the gastric tube re-

gularly at intervals of 2-4 hours. If bleeding is not

stopped, patient is given a paste of Rheum and

Panax orally, and then the esophageal baloon is in-

flated to compress the esophageal wall with the paste

in between. As shown in the table, the mortality rate

has been reduced to about 11-14% in the recent

years. The incidence of post-hemorrhagic coma has

been markedly lowered. Plasma fibrinogen and pro-

thrombin were increased in galactosamine intoxi-

cated rats when treated with Rheum in contrast to

the untreated control.

Death Death Cases Mortality due to due to

shock come

Combined treatment of esophageal varices Western bleeding with western and Chinese treatment

1973-77.6 183 61 (33%) 25 (13.7%) 31 (16.9%) herbal medicine in cirrhotics Combined

Wang BAOEN, et al. treatment 1977-79 89 13 (14.6%) 7(7 .8%) 3 (3 .3%)

Dept Med, Beijing Friendship Hosp, 1980 52 6 (11.5%) 5 (9.6%) 1 (1 .9%) Beijing, China 1981 46 6(13.7%) 4 (8 .7%) 2 (4 .9%)

1982 58 8 (13.7%) 5 (8 .5%) 0 Bebore 1977, we treated massive esophageal p<0.01 p<0.05 p<0.01

bleeding in cirrhotics with the conventional western

I n te rna t i o na l Free Paper Session (3):

Modera to r s : M. P A N C H A N A D A M (India) a n d Shi ro S U Z U K I (Japan)

Clinical spectrum and surgical relief of chronic obstruction of hepatic and suprahepatic segment of inferior vena cava (IVC)--

10 year survey

N. MADANAGOPALAN, V. SOLOMON,

V. JAYANTHI , K. RAGHURAM,

I. KANDASAMY, S. A N N A P O O R A N I ,

A. GAJARAI and M. PANCHANADAM

Dept Gastroenterol, Thoracic Surg, Barnard

Inst Radio ~ Inst Pathol, Madras Med Coil ~ Govt Gen Hosp,

Madras, India

Clinical spectrum, etiopathology, hepatic outflow

and IVC-gram studies in 50 patients with chronic

hepatic and suprahepatic segment obstruction of

IVC with or without hepatic vein (HV) obstruction

seen in 10 years, will be described. Hepatocellular

carcinoma was a common cause of hepatic vein ob-

struction with IVC involvement, but possible throm-


bosis of long segment of hepatic segment of IVC

with or without HV involvement and hour glass con-

striction of IVC just above the drainage of HVs were

also encountered. Such hour glass constriction high

in IVC similar to coarctation of aorta, was observed

in 15. The appearances even suggest a designation

of 'coarctation of IVC. ' Some patients in whom early

surgical repair could have been rewarding presented

after advent of ascites and features of decompen-

sated cirrhosis, making surgery too risky. Even then,

surgery was possible in 10 with such hour glass con-

striction, who presented all features of chronic

Budd-Chiari syndrome. The surgical procedures

used were retro-hepatic cavocaval or cavo-atrial

bypass, using either aortic homograft (3 cases),

plain or ringed P.T.F.E. (Goretex) graft in 6. In one

patient, 'isthmusplasty,' a novel step, offered excel-

lent relief. Special mention will be made of observ-

ance of indisputable histopathological features of

chronic active hepatitis in some with chronic Budd-

Chiari syndrome.

Non-A non-B hepatitis: A health problem in India

Y.K. JOSHI, B.M. GANDHI and

B.N. T A N D O N

Dept Gastroenterol, AIIMS, New Delhi

Non-A non-B (NANB) hepatitis is common dis-

ease in India. Studies at our center have shown that

about 50% of sporadic AVH in adults and 20% in

children is due to NANB infection. Similarly half of

the cases of fulminant hepatic failure and subacute

hepatic failure are due to this infection. Chronic se-

quelae have been observed in 2% of the sporadic

NANB hepatitis. 11 of the 12 epidemics of jaundice

studied by us were due to NANB hepatitis. Disease

predominantely affected the adult population and

had a moderately severe course. No chronic sequelae

has been recorded in one year follow up of the pa-

tients from one of these epidemics. 80% cases of

sporadic NANB hepatitis are suggested to be due to

fecal-oral transmission. Only 15% gave history of

parenteral exposure and 5% had close contact with

jaundiced patients. All the epidemics were water

borne for following reasons. 1) Involvement of large

areas and population. 2) Absence of sewage disposal

system. 3) Improper and inadequate chlorination of

water. 4) Fecal contamination of water. 5) Control

of the epidemic after proper chlorination and other

measures of public hygiene.

These findings are important to plan out the mea-

sures for control of hepatitis in India. Following

measures may prove helpful to reduce the incidence

of NANB hepatitis. 1) Prevention of water con-

tamination with fecal material , 2) Safe water supply

and 3) Health education on personal and environ-

mental hygiene.

The prevalence of non A non B acute

viral hepatitis in Dr. Soetomo

Hospital Surabaya

A. HASSAN, H. GUNTUR, R. SOEMARTO,

S.H. RAHARDJA and W. SOEMARTO

Div Hepato-Gastroenterol, Dept Med, Airlangga Univ, Sch Med,

Surabaya, Indonesia

The aim of the study was to investigate the inci-

dence and the clinical features of patients with non

A, non B acute viral hepatitis.

The diagnosis of acute viral hepatitis A and B can

now be established, because methods for detection

of HAV and HBV markers are now available.

HBsAg was determined by RPHA, while IgM anti-

HAV and anti-HBc by RIA. At present the diagno-

sis of non A non B hepatitis can be made only in-

directly by exclusion of HAV and HBV.

Out of the 108 cases of acute viral hepatitis in Dr.

Soetomo Hospital, 49 cases (39.8%) were HA, 28

cases (25.9%) were HB and 37 cases (34.2%) were

non A non B hepatitis. Of the 9 cases of post-trans-

fusion hepatitis there was only 1 case (11.1%) of HB

and the other 8 cases (88.9%) were non A non B he-

patitis. No HA was found in all 9 PTH cases.

Cl in ica l and immunolog ica l observation on the multisystem manifestations

of chronic hepatitis

Wang BAOEN, et al.

Dept Med, beijing Friendship Hosp, Beijing, China

292 Proceedings of26th Annual Meeting Vol. 20. No. 3

942 cases of chronic hepatitis have been observed

in the past five years. Among them, 126 cases

(13.4%) showed extrahepatic muhisystem mani-

festations. They were: 1) High fever, lasting from

several months to a year. 2 )Jo in t and muscle in-

volvement, including 75 cases (59.5%) of arthralgia

and arthritis, 44 cases (34.9%) of muscular pain. 3)

Skin lesions, showing macropapil lary rashes, urti-

caria, purpura and longstanding itching. 4) Cardiac

and pulmonary lesions consisting of A-V block

(I~176 myocardial damage and interstitial

pneumonitis. 5) Nephritic lesions. 6) Hematologi-

cally, anemia, leukopenia, thrombocytopenia and

aplastic anemia. 7) Beh~et's disease and Sj6gren's

syndrome. Immunological studies showed there was

no difference in incidence of extrahepatic mani-

festations between HBsAg positive and negative

cases.

Serum immune complex detected with methods of

HBsAg specific I.C., PEG, total serum complement

and C3, was positive in 50% of the cases. This sug-

gests that immune complex formation may be in-

volved in the pathogenesis. Immunoglobulins were

elevated, while autoimmune antibodies and cellular

immunity showed no marked change.

Comparat ive studies on HBV markers detection by radioimmunoassay,

RPHA, EIA and C I E P technique

in var ious l iver diseases

Keun-Soo PARK and Bang-Hyun LIU

Dept Intern Med, Coil Med, Pusan Natl Univ, Pusan, Korea

Sera from patients with various liver diseases,

visited to Pusan National University Hospital, from

March, 1977 to June, 1983, were examined for HBV

markers by RIA, RPHA, EIA and CIEP technique.

The results were as follows:

1. HBsAg:

Group with acute hepatitis revealed significant

difference in sensitivity in order of RIA, RPHA and

CIEP. Group with chronic hepatitis showed signifi-

cant difference between CIEP and RPHA, and be-

tween CIEP and RIA, but no significant difference

between RPHA and RIA technique.

In patients with liver cirrhosis, RIA was sensitive

significantly than RPHA, but in hepatoma, there

was no significant difference.

2. Anti-HBs:

Generally RIA revealed higher incidence than

RPHA, but RIA showed significant difference than

RPHA only in group with chronic hepatitis.

3. Other markers:

In group with acute hepatitis, EIA and RIA

showed significant difference for detection of

HBeAg and no significant difference for detection of

Anti-HBc.

HBsAg-associated albumin receptor

in sera of patients with

v i ra l hepatit is

G.B. YAO, L.M. ZHANG, M.F. WU,

S.T. T O N G and Y.F. T I A N

Chn Immunol Res Unit,Jing'an Centr District Hosp, Shanghai, China

The receptor for polymerized human serum albu-

min (PHSA) was studied by hemagglutination

method in 125 cases with viral hepatitis. The geo-

metric mean titer (GMT) of PHSA binding activity

in 50 normal controls was 2.0 + 1.9, and in 50 cases

of acute viral hepatitis A was 4.4 ___ 2.2 (p<0.05).

The GMT titers of PHSA binding in patients with

various HBV infection were as follows: acute hepati-

tis (10) 16.0 _ 7.5, chronic persistent hepatitis (30)

66.6 __ 6.2, chronic active hepatitis (10) 149.4 + 1.7,

chronic asymptomatic carrier (10) 53.8 + 3.2 and

posthepatitis cirrhosis (8) 10.3 _+ 2.7. The correla-

tion coefficient between the PHSA binding GMT

and HBsAg GMT (RPHA) in HBsAg positive sera

was 0.68 (p<0.01). The correlation between PHSA

and HBsAg GMT in cases with HBeAg positive was

also highly significant (r=0.77, p<0.01). The GMT

of PHSA binding activity in 16 cases with HBeAg

positive was 261.8 _ 3.1, and in 58 cases with

HBeAg negative (HBsAg positive) was 20.5 • 5.0

(p<0.01). This study suggests that HBsAg-associated

PHSA receptor may represent a new and useful

virus-associated marker in HBV infection.


I n terna t i o na l Free Paper Session (4):

Modera to r s : W a n g B A O E N (Ch ina ) a n d Kei ichi K A W A I ( Japan)

The action of gastrin, secretin and prostaglandin F2a on lower esophageal sphincter after Nissen

fundopl ica t ion in dog

N. IWAI, H. KANEDA, T. TSUTO,

J. YANAGIHARA and T. TAKAHASHI

Div Surg, Kyoto Pref Univ Med, Kyoto, Japan

This study was done to investigate the action of

gastrin, secretin and prostaglandin on the antireflux

mechanism of the gastroesophageal junction after

Nissen fundoplication.

Nissen fundoplication was performed in 27 mon-

grel dogs under general anesthesia. Esophageal

manometry was done preoperatively, intraopera-

tively, and 2, 4, and 8 weeks after operation.

The injection of 4 r /kg of pentagastrin signifi-

cantly raised the LES pressure both preoperatively

and 8 weeks postoperatively. The LES pressure after

gastrin stimulation increased from 38.1 + 3.5

cmH20 before the operation to 65.0 _+ 8.5 cmH20 2

weeks after operation (P<0.005). However, no sig-

nificant changes occurred in LES length before and

after gastrin stimulation. The injection of 1.5

r / k g / m i n of prostaglandin F2c~ raised significantly

the LES pressure preoperatively. However, the LES

pressure after prostaglandin stimulation did not in-

crease from preoperatively (33.7 + 3,6 cmH20) to 2

weeks postoperatively (33.8 + 7.8 cmH20).

It seems that gastric stimulation to a new mus-

cular sphincter substitute is associated with the anti-

reflux mechanism.

Pyloric sphincter pressure measured with miniature transducer under

fiber-gastroscopy

ZhangJINKUN, et al.

Dept Med, 1st Hosp, Wuhan Med Coil, Wuhan, China

Pyloric sphincter pressure (PSP) was assessed with

a sensitive miniature transducer under gastroscopy.

The mean amplitude of the pyloric high pressure

zone was 21.8 _+ 1.3 m m H g in 50 normal Chinese

adults. The PSP of chronic gastritis (30), peptic

ulcer (38) and gastric carcinoma (6) was diminished,

and those diseases were also found to have pyloric

dysfunction. This data not only present the patho-

physiology and clinical menifestations, but also help

to choose the effective drugs for the treatment. Fur-

thermore, if the PSP 10 m m H g in duodenal ulcer, it

suggests that the protective reflux function of

pylorus is lost.

Carcinoma of gastric stump in China

ZhangJINKUN, et al.

Dept Endos, China Natl Co-operative Group gast Cancer, Wuhan Med

Coil, Wuhan, China

51 cases of gastric stump carcinoma were col-

lected from 18 hospitals throughout China during

the past 9.5 years. There were 47 males and 4 fe-

males. 41 cases (73%) were over 50 years old. The

primary preceding diseases were: gastric ulcer 31,

duodenal ulcer 17 and benign gastric lesion 3 cases.

The types of previous operation were Billorth II 29,

Billorth I 7 and indeterminate 15. The mean inter-

val from the operation to the diagnosis of cancer was

13.8 years. 88% of the present series had a definite

diagnosis by the fibergastroscopy and biopsy. The

rest of the cases were diagnosed by the postoperative

pathologic examination. The X-ray examination

was positive in only 19% of cases. The location of

the cancer was at the anastomotic stoma 45 %, in the

body of stump 36% and in the fundus 19%.

Local irritation following the anastomosis was

considered as a possible cause of malignant trans-

formation. Meticulous care in suturing and the

proper use of resorbable suture material may help

reduce the development of this cancer.

Prevelance of gastric cancer in


Kashmir natives

Hamid A. DURRANI

GI Div, Dept Med, Med CoU, Srinagar, Kashmir, India

Gastric cancer ranks next only to esophageal can-

cer as a common cause of mortality in the native

population of Kashmir valley in the mountains of

north-west India. The incidence is strikingly high

when compared to other population groups in this

country.

The study is a comprehensive analysis of 850 cases

diagnosed in the last five years at one teaching-cum-

referral center. 82% cases were males and 18% fe-

males, 62% smokers and 38% nonsmokers, 60% of

the patients belonged to the age group 41-60 years.

Endoscopic appearances were classified as nodular

mass 47%, polypoid mass 20%, malignant ulcer

10%, infilterative lesion 8%, rounded tumor mass

8%, linitis plastica 5% and early gastric cancer 2%.

On histopathological typing 79% lesions were

adenocarcinomas, 8% mucoid carcinomas, 8%

leimyosarcomas, 4% schirrous carcinomas, and 1%

reticulum cell sarcomas.

The possible etiological factors peculiar to the

dietary constituents are singled out. Genetic factors

may have an important role in the etiology of gastric

cancer in this high risk group.

Small bowel changes in chyluria

J.P. GUPTA, A.K. JAIN, A.K. GOVIL

and V.N.P. T R I P A T H I

Div Gastroenterol, Inst Med Sci, Banaras Hindu Univ, Varanasi-5, India

Chyluria is one of the common manifestations of

filariasis. Although there is retroperitoneal lymph-

angiectasia with lymphatic varices in chyluria, the

involvement of intestinal lymphatics and its adverse

effects on intestinal structure and function remain

unknown.

Sixteen subjects of chyluria (urine +ve for chyle)

with repeated stool examination negative for ova

and cyst were studied. In them urinary excretion of

d-xylose, faecal fat excretion, gastrointestinal (GI)

protein loss (using RIHSA), serum proteins with

A:G ratio and lipidograms were done. Barium con-

trast jejunal radiography and jejunal biopsies were

also studied. The majority (88%) had no GI symptoms. Serum

lipid profile and faecal fat excretion were normal in

all. Three fourth subjects had reduced serum albu-

min with normal globulins. RIHSA studies revealed

decreased half life in chylurias compared to con-

trols. Jejunal biopsy showed marked to moderate

lymphangiectasia in three fourths with grade II to

grade III villous changes in 40%.

Thus, although varying degree of intestinal lym-

phangiectasia was present in the majority of chylu-

rics and was associated with villous changes in more

than third but biochemical evidence of malabsorp-

tion was absent.

Cl in ica l appl icat ion of "Purgation Method" in t reat ing digestive

diseases and its mechanism

Wang BAOEN, et al.

Dept Med, Beijing Friendship Hosp

"Purgation method" of Chinese traditional medi-

cine has been used in combination with western

medicine in treating digestive diseases, such as acute

pancreatitis, cholecystitis, liver abscess, bacillary

dysentery and upper G-I bleeding. Good clinical re-

sult was observed, especially in regard to the subsi-

dence of fever and reduction of toxic symptoms.

Animal experiment showed that the antipyretic

effect is at tr ibuted to the chief component of the

purgative mixture, Rhubarb. Further experiment

revealed that rhubarb could significantly reduce the

activity of prostaglandin E in the C.S.F. drained

from the third ventricle near the thermoregulation

center (P<0.001). The lowering of anal temperature

is proportionally correlated to the reduction of PGE

in the C.S.F. (r=0.72, P<0.01). The purgative mix-

ture also increases the phagocytic function of PMN

and the activity of total serum complement. It can

also counter-act endotoxin in the blood. The hemo-

static effect is due to the local effect of Rheum as

well as some change of the coagulation factor.

Alteration in human colonic muc in


b i n d i n g to the lectins in the diagnosis of colonic

adenocarcinoma

Z.R. SHI, D.Y. YIN and F.Q. WEN

Dept Med, 2nd Aj~liated Hosp, Hunan Med Coll, Changsha, China

Changes in the pa t te rn of colonic mucin secretion

has been described in colon cancer. In this study,

the b ind ing of F ITC-PNA (peanut agglut inin) and

FITC-DBA (dolichos biflorus agglut inin) to the

muc in in the colorectal ca rc inoma (Ca), t ransi t ional

mucosa (Tin), normal colonic mucosa (Nm), adeno-

matous polyps (Ap), villous adenomas (Va) and

mult iple adenomas (Ma) were studied by using fluo-

rescence microscopy. The b ind ing capacity was

scored as 0, no; 1+ weak; 2+ modera te ; 3+ strong

fluorescence. The result is listed on Tab le 1.

Table 1. Bindingof FITC-PNA and FITC-DBA to different kind of colonic mucosa

Ca Tm Nm Ap Va Ma n=25 n=25 n=25 n=20 n=10 n=5

1+ 8 2+ 3 1+ 5 0 4 1+ 5 2+11 3+ 22 2+ 3 1+ 4 3+ 6 3+ 12 2+ 2 0 12 0 25 0 20 0 2 0 4 1+ 8 1+ 3 1+ 1 2+ 5 2+ 5

DBA 0 18 1+ 4 2+ 3

PNA 1+ 3 2+ 8 3+ 14

An exposed carbohydra te s t ructure in the secreted

mucin was found in h u m a n colonic carc inoma

which was not present in no rma l colonic mucosa. It

may be of benefit for the diagnosis of colorectal car-

c inoma.

Direct vision of the pancreas wi th

laparoscope

J inkun ZHANG

Dept Med, 1st Hosp, Wuhan Med Coil, Wuhan, China

We, with supragastr ic approach th rough the

lesser omentum, inserted the endoscope into the

lesser peri toneal sac for direct vision of the pancreas.

Twelve cases were examined and ten of them were

successfully done. T h e diagnoses were: Three nor-

mal , four chronic pancreat i t is , one pseudocyst, one

cancer of pancreat ic body and one cancer of pancre-

atic head. All diagnoses were supported by the

ERCP, B-type ul t rasonography, and BT-PABA test,

etc.

This paper discusses the endoscopic technique for

direct vision of the pancreas and how to perform the

laparoscopic examina t ion with the GIFP3 and its

accessories such as cold l ight supply, biopsy forceps

with UES generatory and photography.

In terna t iona l Free Paper Session (5):

M o d e r a t o r s : D i n g - S h i n n C H E N ( T a i w a n ) a n d Yasuo E N D O ( J a p a n )

Correlated c l in ica l , h i s topa tho log ica l ,

HBV and AFP studies in primary carcinoma liver and speculations

on possible additional role of non -ed ib l e oi l i n

pa thogenes is

M, PANCHANADAM, V.C. BALASUBRAMANIA,

P. SARE, S.P. THYAGARAJAN,

S. MAHADEVAN and S. D R A V I D A M A N I

Inst Pathol, Microbiol and Animal Experimental La b, Madras Med

Coll, Madras-3, India

This paper presents clinical and histopathological

features of over 300 cases of p r imary liver cancer

over last 14 yrs. This large n u m b e r suggests increas-

ing prevalence in last 2 decades. Special features

are, a) 40% are below age 40, b) 75% have sympto-

mat ic illness <6 too, c) large propor t ion are non-

alcoholic. The initial HBsAg (+) was 31%, but it in-

creased to 60% with ELISA. AFP was <200 n g / m l

in 20% of those who were (+). Ground nut oil was

the ma in cooking oil in most patients. It became es-


sential to explore possible role of con taminan t s and

adulterants of oils for the apparen t increase of HCC,

particularly the short dura t ion illness in younger age

group. The impor t an t change over last 2 decades

has been the shift from cattle operated rotaries for

ground nut oil extract ion to huge electric rotaries

which at times operate for extract ing nonedib le oils

such as cashew nut shell l iquid (CNSL). If potent ial-

ly toxic oil were to con tamina te edible oil, more so if

already rich in mycotoxins, and consumed by people

at high risk with HBV infection, it may speed up

evolution of HCC. Prel iminary studies in rats have

shown hepatic pa renchymay and bile duct changes,

and unusual f ibroadenoma breast, in animals fed

with ground oil mixed with minimal amounts of

CNSL. Animal studies with pure ground nut oil and

commercial oil mixed with CNSL will also be pre-

sented.

Primary hepato-cellular carcinoma

Pranesh NIGAM, A.K. A G R A W A L and

K.K. K A P O O R

Dept Med, BRD Med Coll, Gorakhpur-273013,

U.P., India

Fifteen patients (mean age=58.6 yrs, sex

ra t io=4 .5 : l ) were studied for the clinical profile. As-

sociated cirrhosis was seen in 5 males who deterio-

rated for no obvious reason. Usual presenta t ion of

malaise, abdomina l fullness, anorexia, const ipat ion

etc, was mainly seen. Dull acheing type of pain over

epigastrium, r ight upper quad ran t and back was

present in all of them which was severe in one case

with friction rub over epigastr ium. Mild to moder-

ate jaundice was seen with nodular ha rd liver. Left

lobe of liver was exclusively involved in 2 cases. Arte-

rial m u r m u r over r ight hypochondr ium was audible

in 4 cases. Ascites was present in 6 cases and ascitic

fluid revealed high prote in (4.6 gms%), lactic de-

hydrogenase (108 IU) and mal ignan t cells (3 cases).

Massive blood t inged r ight sided pleural effusion de-

veloped in a lady, and painful gynecomastia was

present in one case. Hypoglycemia (fasting 50 m g % ,

PP 90 m g % blood sugar) noted in 4 cases. Serum

cholestarol was raised (382 m g % ) in 3 cases, Serum

alk. phosphatase was markedly raised (102 IU) with

ra ised s e r u m t r a n s a m i n a s e s . Ery throcy tos i s

(Hb.=16.2 g ins%) was seen in one case. Histology

revealed smaller t umor cells resembling normal liver

cells with compact finger like processes or solid tra-

biculae. Max imum survival=l year.

Diagnostic value of fine needle aspiration: A prospective study

L. LESMANA, H. SIDHARTA,

W. M A R W O T O , N. AKBAR, A. SULAIMAN

and H.M. SJAIFOELLAH NOER

Dept Med and Anatomic Pathol Univ

o f Indonesia, Jakarta, Indonesia

This study was carr ied out to assess the accuracy

of fine needle aspirat ion (FNA) in focal hepat ic

lesion.

Dur ing an eight months period (August 1, 1983 to

April 30, 1984) FNA under ultrasonic guidance was

performed on 60 pts in whom US revealed a hypo or

hyperechoic lesion(s). A real t ime sector or l inear

scanning with a c o m m o n t ransducer were used for

this purpose. The aspirated mater ia l was stained ac-

cording to Papanico laou technique for cytological

examinat ion .

Results: Correct diagnosis of FNA was found in 53

of 60 pts (88.3%); 44 with liver cancer (LC) and 9

with abscess of the liver (AL). False negative diagno-

sis was ob ta ined in. 3 pts (5%) with pr imary LC. In-

conclusive diagnosis was found in 4 pts (6 .7%) one

pt proved to have secondary LC on liver biopsy, in 2

pts a follow-up US showed no growth of the lesion

and the r ema in ing one pt refused to undergo fur ther

examinat ions. Complicat ions were not noticed in

this study.

Conclusion: Fine needle aspirat ion is a safe

method and has a high diagnostic value in focal

hepatic lesion.

Serodiagnosis of hepatic neoplasia

J .P. G U P T A , B.V. T A N T R Y and

A . K . J A I N

Div Gastroenterol, Inst Med Sci, Banaras

Hindu Univ, Vanarasi-5, India

Serological diagnosis of tumors is a recent de-


velopment and has been used in various malignancies. Amongst various tests LAI (leukocyte ad- herence inhibition) is a reliable, rapid and simple

technique. In the present study, LAI test was done in patients

with established primary and secondary carcinomas of liver using specific primary and secondary liver carcinoma antigens. Six healthy subjects and 16 patients of non-malignant hepatic diseases (e.g. cirrhosis, viral hepatitis and amoebic liver abscess) served

as controls. Positive LAI test was obtained in all the six cases

of HCC (hepatocellular carcinoma) using HCC antigen. There was no false positive among both the

control groups whereas 2/21 cases of secondary car-

cinoma of the liver gave false positive tests with HCC

antigen. Using sec. adenocarcinoma liver antigen, 19/21

cases of sec. carcinoma of the liver gave positive tests. There were two false positives among non- malignant liver disease controls and 1 (out of 8) in

HCC. Thus, in the present study LAI test was found to

be helpful in diagnosis of hepatic neoplasia.

Sonographic evolution of hepatocellular carcinoma in its early stage

J.C. SHEU, D.S. CHEN, J.L. SUNG, C.N. CHUANG, P.M. YANG and

J .T. LIN Dept Intern Med, Natl Taiwan Univ Coll

Med, Taipei, Taiwan R. O. C.

To study the sonographic evolution of hepatocellular carcinoma (HCC) in its early stage, 26 HCCs smaller than 5 cm in 24 patients, were ob-

served regularly by real-time ultrasound for a period of 90 to 691 days. In the beginning, 21 tumors were hypoechoic, 2 isoechoic, and 3 diffusely hyperechoic. On follow-up, 2 of the 21 initially hypoechoic HCCs remained in the same echodensity, 12 increased in internal echoes but were still hypoechoic; 1 became isoechoic, 4 changed to hyperechoic pattern; and the remaining 2 shifted from hypo- to iso- and then to hyperechoic pattern. The 2 initially isoechoic HCCs also gained echogenecity

and became hyperechoic. By contrast, the 3 initially hyperechoic HCCs kept on the same echofeatures. Those "acquired" hyperechoic HCCs were inhomo- geneous in echodensity and larger in size whereas the

3 origionally hyperechoic HCCs were homogeneous

and smaller. It is concluded that most small HCCs evolve progressively from hypoechoic to iso- then to

inhomogeneously hyperechoic pattern as they grow; a few HCCs get their diffusely high echogenecity

early since their beginning and remain in the same feature thereafter.

Prognostic histologic features of resected hepatocellular

carcinoma in Taiwan

H.C. HSU, J.C. SHEU, Y.H. LIN, D.S. CHEN, C.S. LEE, R.P. BEASLEY and

L.Y. HWANG Dept Pathol, Intern Med, and Surg,

Natl Taiwan Univ, and Univ of Washington Med Res Unit,

Taipei, Taiwan R. O. C.

This study analyzed the morphologic features and

their prognostic significance in 83 surgically resected hepatocellular carcinoma (HCC) which had been

followed up from 10 months up to 4 years, or until recurrence occurred: 44 were of <5 cm in diameter

and 39 were larger. Although small HCC had a higher incidence of cirrhosis (72.7%) than large HCC (43.6%), both had high prevalences of tumor capsule (86.4% and 84.6%, respectively), and HBsAg positivity (93.2% and 87.2%, respectively). However, small HCC had a higher HBsAg positivity in the tumor (40.9%) than large HCC (10.3%), P<0.003. In small HCC, the capsule was more frequent in liver with cirrhosis (31/32) than without (7/12), P<0.001. This comparative study demonstrated that the utmost important histologic para- meter influencing the tumor recurrence was the liver invasion for both small (P<(0.001) and large HCC (P<0.05). For small HCC, the extracapsular invasion also posed a worse prognosis than without,

P<0.05. The better prognosis of small than large HCC was attributed to the lower incidences of liver invasion (P<0.001), portal vein invasion (P<0.01),


and satellite nodule (P<0.01). The tumor recur-

rence did not correlate with the tumor grading,

presence of clear cells, or liver cell dysplasia. These

findings indicate tha t the prognosis of HCC may be

predetermined by the HCC itself.

Metastatic malignancy of liver

Pranesh NIGAM, K.K. K A P O O R and

A.K. G U P T A

Dept Med, BRD Med Coll, Gorakhpur-273013, India

Thir ty cases (25 males, 5 females, mean age=62.5

+ 5.6 yrs) of metas ta t ic mal ignancy of liver were

studied for the clinical profile. Majority of them

(66.6%) presented with non-specific symptoms of

weakness, weight loss, fever, sweating and loss of ap-

petite. Pain over r ight hypochondr ium was sharp

and in te rmi t ten t in 16,6% of cases. Tende r hepato-

negaly with mul t ip le ha rd nodules were observed in

53 .3% of cases. Jaundice was deep in 13.3% and

mild in 6 .6% of cases. Friction rub over liver area

was audible in 10% and splenomegaly in 50% of

cases. Right supra-clavicular lymph glands were en-

larged in 6 .6%. Oedema over leg was seen in 33 .3%

of cases. Blood stained pleural effusion and ascites

was seen in 6.6 % and 13.3 % of cases respectively.

Hyperb i l i rub inaemia was present in 23 .3% of

cases. Serum alk. phosphatase, lactic dehydrogenase

and t ransaminases levels were raised in all of them.

Ratio of lactic dehydrogenase in ascitic fluis and

serum was more t han 3 in 13.3% of cases. Trea t -

ment of these cases remained highly unsatisfactory

with survival of 3-6 months from the diagnosis

established.

Proceedings of the international session of the 26th annual meeting October 18–20, 1984—Chiba,...

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Transcript of Proceedings of the international session of the 26th annual meeting October 18–20, 1984—Chiba,...