Critical role of bile acid (BA) in the cellular entry and permissiveness of Hepatitis B virus in vitro

Hong‑Jai Lee,Bo‑Young Shin,Jae‑Seung Moon,Ailyn Fadriquela,Selikem Abla Nuwormegbe,Chun‑Chang Ho,Jin‑Su Shin,Jee‑Sang Yoon,Sang‑Kyou Lee,Soo‑Ki Kim 대한독성 유전단백체 학회 2020 Molecular & cellular toxicology Vol.16 No.3

Background Lack of universal replication system for hepatitis B virus with narrow host range and organ tropism has hampered to uncover the pathogenesis of HBV. Previously, we reported the essentiality of humoral milieu and its components toward HBV and hepatitis C virus survival/viability in vitro. Of these components, the precise role of enterohepatic humoral milieu such as bile acid (BA) on HBV cultivation in vitro and in vivo is unknown. Objective We explored whether BA, specifically taurochenodeoxycholic acid (tCDCA) would directly regulate the viral DNA and surface antigen expression of HBV in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. Result We found that higher concentration of taurochenodeoxycholic acid (tCDCA) is able to preserve the genomic stability of HBV in cell-free DMEM, showing higher the surface antigenicity than taurocholic acid (tCA). In line, we found that in vitro cell culture condition (100 μmol/L of tCDCA coupled with 1 × 108 g e/mL HBV) would be optimal for HBV entry into target cells. Using this, human (HepG2, Huh7), and rodent (Hepa1c1c7, H4-II-E) hepatoma cell lines were infected by HBV, as evidenced by the presence of HBV biomarkers (HBsAg, and HBV DNA in culture supernatant, as well as HBcAg in cell). Further, cellular entry test revealed that HBV is able to infect 12 different non-hepatic cell lines regardless of species, and organ/tissue, consequently reproducing progeny as confirmed by HBV biomarkers. Last, reinfection test showed that the progenies of HBV from immortalized HepG2, and Hepa1c1c7 cells are able to enter into each or vice versa naïve HepG2, and Hepa1c1c7 cells with or without BA. Conclusion This study demonstrates that enterohepatic humoral milieu such as BA, specifically tCDCA would directly regulate HBV DNA and its surface antigen expression in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. This is the first note to render HBV permissive to human or rodent hepatic and non-hepatic cells via sole manipulation of humoral milieu, thus establishing the platform for in vitro robust replication system of HBV.

The HBV DNA Amounts in Serum Have No relationship with ALT level and Hetergeneous Population Coexits in Chronic Hepatitis B Virus Infection

Chun, Yoon-Keun,Ha, Joo-hun,Hong-Jung-Woo,Oh, Soo-Myung,Kim, Sung-Soo 경희대학교 동서의학연구소 1999 INTERNATIONAL SYMPOSIUM ON EAST-WEST MEDICINE Vol.1999 No.1

Yoon-Keun Chun¹,Joohun Ha□Hong-Jung Woo□, Soo Myung Oh□,Sung Soo Kim□ ¹Department of Molecular Biology, College of Medicine,²Department of Surgery, college of Medicine,³Department of Internal Medicine, College of Oriental Medicine,and ⁴East-Weat Medical Reserch Institute,Kyung Hee University, Seoul, Korea. The HBV DNA Amounts in Serum Have No relationship with ALT level and Hetergeneous Population Coexits in Chronic Hepatitis B Virus Infection. Proceedings of International Symposium on East-West Medicine, Seoul. 212-230, 1999. -Hepatitis B is caused by hepadnavirus. Hepatitis B virus replicates through 3.5kb pregenomic RNA intermediate which is regulated by core promoter. Pathogenesis of hepatitis B virus has been bilieved the result of host immune response. But recently many studies have reported that high level of viral replication caused by mutation in core promoter might result in severs hepatitis. But these studies were performed in vitro, not in vivo. So there is yet debate about which factor, viral of host factor, is more important in pathogenesis of hepatitis B virus. So we measured real viral replication level in 204 chronic hepatitis B patients by quantifying HBV DNA from sera by our novel PCR-based more sensitive method, and compared these results with ALT level measured from same sera, which indicates liver cell damage. Surprisingly there are no significant correlation between HBV DNA quantity and ALT level. Then we cloned core promoter region. In SSCP, we found that many viral mutants coexist in one patient. Base on SSCP result, we chose main viral core promoter type in each patients, which is thought to determine overall viral replication level in this patient. Main type of core promoter region of each 41 patients were directly sequenced. And with these we measured promoter activity by luciferase assay system and compared promoter activity with on another. We found tha there were some differences in promoter activity according to core promoter sequences. And we constructed replication-competent viral constructs with core promoter from 41 patients and Transfected these into HepG2 cell and measured HBV DNA by southern blot. There were also differences in HBV DNA quantity according to core promoter sequences. On these all results we investigated correlation between the effect of HBV core promoter on viral replication in vitro and HBN DNA quantity, ALT level from sera of each patients. We found there is no significant correlation among them. As a result, we concluded that in determining severity chronic hepatitis B patients, host factors of each patient is more important rather than replicative activity of virus itself.

서울의 Penicillinase Producing Neisseria gonorrhoeae 발생빈도(1998)

김재홍,김준호,반재용,이정우,황성주,정준규,정성태,강진문,조흔정,홍창의,정혜신,이한승,김이선,이봉길,이종호,선영우,한기덕,윤성필,이성훈,안종성,박석범,문승현,조항래,김형섭,류지호,황재영,박준홍,손상욱 한양대학교 의과대학 2001 한양의대 학술지 Vol.21 No.1

In recent years, gonorrhea has been pandemic and remains one of the most common STDs in the world, especially in developing countries. For the detection of a more effective therapeutic regimen and assessing the prevalence of Penicillinase Producing Neisseria gonorrhoeae(PPNG), we have been trying to study the patients who have visited the Venereal Disease Clinic of Choong-Ku Public Health Center in Seoul since 1980 by menas of the chromogenic cephalosporin method. In 1998, 93 strians of N. genorrhoeae were isolated, among which 60(64.5%) were PPNG. The prevalence of PPNG in Seoul, which had been decreased to 39% in 1996 after a peak of 74.3% in 1993, is increased to 64.5% in 1998.

서울의 Penicillinase Producing Neisseria Gonorrhoeae 발생빈도(1996)

김재홍,황동규,전재홍,김윤석,김중환,김용준,이창균,임동진,김현수,조창근,김경문,박상훈,전우형,김희성,이호정,차명수,김갑형,김형석,김석우,황지환,박병순,권오상,이민수,송기훈,성소영,이인섭,부태성 대한화학요법학회 1999 대한화학요법학회지 Vol.17 No.2

Background : In recent years, gonorrhea has been panedemic and remains one of the most commom STDs in the world, especially in developing countries. Objective & Methods: For the detection of a more effective therapeutic regimen and assessing the prevalence of PPNG, we have been trying to study the patients who have visited the VD Clinic of Choong-Ku Public Health Center in Seoul since 1980 by means of the chromogenic cephalosporin method. Results: In 1996, 139 strains of N. gonorrhoeae were isolated, among which 53(39.0%) were PPNG. Conclusion: Our results suggests that after a peak of 74.3% in 1993, the prevalence of PPNG in Seoul is gradually declining.

정신분열병에 대한 리스페리돈의 효과 및 안정성

이민수,김용구,김영훈,연병길,오병훈,윤도준,윤진상,이철,정희연,강병조,김광수,김동언,김명정,김상훈,김희철,나철,노승호,민경준,박기창,박두병,백기청,백인호,손봉기,손진욱,양병환,양창국,우행원,이정호,이종범,이홍식,임기영,전태연,정영조,정영철,정인과,정인원,지익성,채정호,한상익,한선호,한진희,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.1

연구목적 : 본 시험의 목적은 임상시험 시작전에 연구자들을 대상으로 PANSS Workshop을 통하여 PANSS, ESRS에 대한 국내에서의 표준화 작업을 구축하고 새로운 정신병 치료제인 리스페리돈의 효과와 안정성을 재확인하여 리스페리돈 사용에 대한 적정화를 이루는데 있다. 연구방법 : 1996년 4월부터 1996년 9월까지 국내 39개 대학병원 정신과에 입원중인 혹은 증상이 악화되어 입원하는 정신분열병 환자 377명을 대상으로 다시설 개방 연구를 시행하였다. 1주일간의 약물 배설기간을 가진후, 리스페리돈을 8주간 투여하였고, 기준점, 1주, 2주, 4주, 그리고 8주후에 평가되었다. 용량은 제1일에는 리스페리돈 1mg씩 1일 2회, 제2일에는 2mg씩 1일 2회, 제3∼7일에는 3mg씩 1일 2회 투여하였다. 이후 환자의 임상상태에 따라 임의로 증량할 수 있으며, 최대 일일 16mg을 초과하지 않도록 하였다. 추체외로 증상을 조절하기 위한 투약을 허용하였다. 임상증상 및 부작용의 평가는 PANSS(Positive and Negative Syndrome Scale), CGI(Clinical Global Impression) 그리고 ESRS(Extrapyramidal Symptom Rating Scale)을 사용하였다. 연구결과 : 377명중 343명(91%)이 8주간의 연구를 완결하였다. 치료 종결시점인 8주후 PANSS 총점수가 20% 이상 호전된 경우를 약물 반응군으로 정의할때, 약물반응군은 81.3%였다. 리스페리돈에 반응하는 예측인자로는 발병연령, 이전의 입원 횟수, 유병기간이 관련 있었다. 리스페리돈은 1주후부터 PANSS양성, 음성, 및 일반정신병리 점수상에 유의한 호전을 보여 효과가 빨랐다. CGI의 경우도 기준점에 비해 1주후부터 유의한 감소를 나타내었다. ESRS의 경우, 파킨슨 평가점수는 기준점과 비교해 투여 1주, 2주, 4주후 유의하게 증가되었다가 8주후 기준점과 차이가 없었다. Dystonia 평가점수는 1주후만 유의한 증가를 보였으며, dyskinesia 평가점수는 유의한 차이가 없었다. 혈압, 맥박수의 생명징후 및 일반 혈액학 검사, 생화학적 검사, 심전도 검사에서 유의한 변화는 없었다. 결 론 : 이상의 다시설 개방 임상 연구를 통해 리스페리돈은 정신분열병 환자에서 양성증상뿐만 아니라 음성증상 및 전반적인 증상에도 효과적인 것으로 사료된다. 보다 명확한 평가를 위해서는 다른 항정신병약물과의 이중맹검 연구가 필요할 것으로 생각되며, 또한 장기적 치료에 대한 평가도 함께 이루어져야 하겠다. Objective : The purpose of this study was to investigate the efficacy and safety of risperidone in the treatment of Korean schizophrenic patients. Method : This multicenter open study included 377 schizophrenic patients drawn from 39 university hospitals. After a wash-out period of 1 week, the schizophrenic patients were treated with risperidone for 8 weeks and evaluated at 5 points ; at baseline, and 1, 2, 4 and 8 weeks of treatment. The dose was increased from 2mg/day(1mg twice daily) to 6mg/day(3mg twice daily) during the first week and adjusted to a maximum of 16mg/day over the next 7 weeks according to the patient's clinical response. Medication to control extrapyramidal symptoms was permitted. The psychiatric and neurological status of the patients was assessed by PANSS, CGI, and ESRS scales. Results : 343(91%) of 377 patients completed the 8-week trial period. Clinical improvement, as defined by a 20% or more reduction in total PANSS score at end point, was shown by 81.3% of patients. The predictors of response to risperidone were associated older age, shorter duration of illness, fewer previous hospitalization. Risperidone had rapid onset of action ; a significant decrease of the total PANSS and three PANSS factor(positive, negative, general), and CGI was already noticed at the end of first week. For the ESRS, parkinsonism rating scores were significantly increased until week 4 comparing with baseline. Dystonia rating scores were significantly increased until week 1, and dyskinesia rating scores were not significantly changed during the study. Laboratory parameters including vital sign, EKG, hematological, and biochemical values showed no significant changes during the trial. Conclusions : This study suggests that risperidone is generally safe and effective against both the positive and negative symptoms in our group of patients.

親環境農業을 위한 有機農業의 作付體系 確立에 관한 硏究

홍성찬,강희경,윤영상 공주대학교 자원과학연구소 2000 資源科學硏究論文集 Vol.8 No.-

This study was carried out to produce safe agricultural product and to improve the soil environment of crop cultivation using the reasonable cropping system in organic farming to the case study of organic farmhouse in Chungnam Hongdong area for sustainable agriculture. In the ratio of farming forms in these organic farmhouses, paddy field-upland field-greenhouse form, paddy field-upland field form, crop-livestock form and only greenhouse form were 23.8%, 23.8%, 23.8% and 19.0% respectively. In the greenhouse area of organic farmhouse, the 50% of total greenhouse number were 600~1,000 pyung(200~330a) and the 40% of total were 1,000~1,500 pyung(330~500a). In the area of upland field, the 80% of total upland field number were 500~1,000 pyung(167~330a). In the cropping systems of surveyed farmhouses, the number of crop-combination patterns were twenty-seven and the 96.3% of those was combined with vegetable. Vegetable-vegetable pattern, cereal crops-vegetable pattern and root and tuber crop-vegetable pattern were 44.4%, 18.5% and 14.8% respectively. Leguminous crop combined pattern was 7.4%, but patterns combined with barley or wheat did not exist. Among the vegetable combined patterns, leaf vegetables combined pattern, fruit vegetables combined pattern and root vegetables combined pattern were 66.7%, 31.0% and 22.2% respectively. Among the vegetable-vegetable patterns, the portion of fruit vegetables-leaf vegetables pattern was the highest and that of root and tuber vegetables-root and tuber vegetables pattern was the lowest. Among the twenty-seven crop-combination patterns, unreasonable crop rotation pattern was 50% over and especially the almost crop-combination pattern of greenhouse was unreasonable to crop rotation pattern. The farmhouse that control the amount of compost application by means of periodical soil analysis did not exist in this area.

요골동맥을 이용한 관동맥 조영술의 유용성

윤정한,이승환,이한요,김장영,김일회,최윤종,이형준,이명옥,김승년,황성오,홍인수,최경훈 대한순환기학회 1998 Korean Circulation Journal Vol.28 No.10

消癌去痰益氣湯의 cyclophosphamide 부작용 형성 억제효과

홍종희,류기원,류봉하,윤상협,김진성 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2002 東西醫學硏究所 論文集 Vol.2002 No.-

Object : It is well known that modern chemotherapy against cancer has side effcts to a living body, especially hemopoietic and immunologial disfunctions. However, there are no effctive ways to reduce them.Recently, traditonal Korean herb medicine has been repoted to have some biological modifying responses. Therefore, we hypothesized that additional application of herb medicine during chemotherapy is more effective to reduce its side effects. While we were studying the effects, we have observed the inhibitory effect of Soamgudamikgitang on formation of side effects derived from Cyclophosphamide, it has been used in clinical practice at Kyung Hee Medicd Center.Methods : We injected 200㎎/㎏ of Cyclophosphamide, one time, to an experimental group, consisting of ten mice. We divided them into eight groups: normal, CPX, SAKT 2㎎, SAKT 10㎎, SAKT 50㎎, SAKI 2㎎+CPX, SAKT 10㎎+CPX, SAKT 50㎎+CPX. We injeted Soamgudamikgitang seven days, five days, three days, and one day before we injected CPX. One day, three days, and five days after CPX injection, we injected Soamgudamikgitang again and then killed all the mice.The parameters determined in this experiment were daily body weight liver and spleen weight, RBC, WBC, and platelet for hemopoietic dysfunction and AST, ALT for hepatotoxicity, BUN, creatine for renal toxcity, Iymphocyte proliferation activity and lymphocyte subsets for immunological toxvity.Results : We have found that Soamgdamikgitang has inhibitory effects on the formation of Cyclophosphamide 's side effects. Significant differences between the group, which contained Cyclophosphamide, and the other group, which contains Cyclophosphamide and 2, 10, 50㎎ of Soamgudamikgitang respectively were observed. Platelets(2㎎ of Soamgudamikgitang, p<O.05 ;10㎎, p<0.Ol ;50㎎, p<0.001), liver weight(SOw p.001), spieen weight(10mg, p<0.05), AST(all groups, p<0.01), ALT(2㎎, p<001 ;10㎎, p<O.05 ;50㎎, p<0.01), BUN(2㎎, p<0.01 ;50㎎, p<0.05). Althought immunological in both lymphocyte proliferation and its subsets were not observed, which shows that Soamgudamikgilang has a strong effect on T cell activities Conclusions : From the above results, we can expect that the combined therapy of Soamgudamikgitang and Cyclophosphamide is more effective for treating cancer Patients.

地域經濟活性化를 위한 農村市場開發에 관한 硏究 : 忠南 禮山地域市場을 中心으로

尹畯相,李武鉉,申尹撤,洪性贊,朴奉圭 公州大學校 産業開發硏究所 1994 産業開發硏究 Vol.2 No.-

Since the first year economic development plan, imblanced development of national territory during the economic development process has aggravated remarkable disparity between urban and rural areas. For this reason, this study introduced the development of rural market as one of the development strategies of regional economic to correct the imblance and relieve regional disparity. The purpose of this study were to look into the present situation of traditional rural market and to analyze the problems for the rural development in view of the current road situations in the rural areas and status of marketing facilities in market center and to measure the degree of rural area people's satisfaction to the rural market and to suggest the development plan of activating for rural market center. To collect data, the Yesan Gun rural market were selected and a total of 591 residents and 178 merchants were sampled as final subjects of survey. Interview and/or questionnaire method was the main method of data collection. The major findings of this study were as follows: 1) The population of Yesan Gun has decreased 2.85 persons per year from in 1984, but the number of vehicles were increased 2.85 times than in 1989. And the parking capacity of Yesan Eup was only 129 vehicles which were below of the total vehicles of Yesan Gun area (14,352). The number of Yesan area markets were 8 markets which consisted of 1 permanent market and 7 periodic markets. But as the economy has been developed, the conditions of rural markets have been changing. That is, the role of periodic market tends to be weakened with the growth of large cities, reinforcement of permanent markets, development of traffic facilities, large production, preferring high quality goods. 3) Generally, the people of Yesan Gun were attached for purchasing the necessaries of life and comodities and farming materials to ① Yesan Eup ② Seoul ③ Chonan Shi ④ Hongsung gun ⑤ Daejeon Shi. 4) The degree of community people's satisfaction to the town market presented 26.6% which meaned higher than dissatisfaction(9.5%). And the degree of people's inconvenience of accessibility to market center by transferring of bus terminal presented 64.2% which meaned higher than average. 5) The degree of people's dissatisfaction of traffic system in Yesan gun area presented followings: the road networks(61.5%), parking facility(68%), bus route(54%). 6) By the survey of merchants of Yesan gun area, on the average, it showed followings: the size of store was 6-10 pyeong(40%), the type of enterprise was retail dealer(70.9%), and purchasing place was wholesale dealer from Seoul(51.8%). 7) And they thought that Yesan town market were too small(70.8%), and the diversifying of goods was not enough(35.5%). 8) The merchants of Yesan area showed that the degree of into venience to traffic system was 72.7% and its to parking facility was 83.3%. And they complained that community administration agency have not tried to activate market(79.1%). Based on the results of this study, the following recommendation were made for activating regional economy and community market. That is, it is to strengthen the economic function in community market so as to suffice the economic and social activities for settlement area as their living space. And it is necessary to improve the interrelationship of the central functions. 1) So, the accessibility between market center and its surrounding areas must be rised. ① the enforcing preferentially to pave the bus terminal and market center to link two place. ② the running of shuttle-bus two place every regular market day. ③ the widening and connecting the road to rise accessibility between market center and its surrounding area. ④ the enforcement of parking facility around market center. ⑤ the constructing marketing centered area as intergrated body of various marketing facilities in the adjacent site is the centered area planned to achieve efficient marketing. 2) The modernization and specialization market center facilities. 3) The strengthening of drawing surrounding area people into Yesan market center. So, the following recommended: ① the development of special products market ② inducing special projects ③ the development tour and leisure course drawing industrial facilities. 4) And it needs to enforce community people's local patriotism and to support community markets by administration agency.

쇄골 중간부 골절후 불유합 및 지연 유합의 원인

윤정로,심재익,김택선,이성종,김영배,김학준,안국환,장재영,홍명표 대한골절학회 2002 대한골절학회지 Vol.15 No.4

쇄골 중간 1/3 부위의 골절은 치료 결과가 양호하여 보존적 치료가 원칙적으로 받아 들어지고 잇으며 불유합에 대한 빈도도 낮은 것으로 되어 있다. 그러나 최근 전위 및 단축의 정도가 심할 경우 높은 불유합의 결과와 불량한 임상 결과를 보고하고 있어 이런 예에 대해선 수술적 치료를 권장하고 있다. 저자들은 단축 및 전위의 정도와 불유합의 연관성을 알아보고자 하였다. 연구 대상 및 방법 : 1993년 2월부터 2002년 1월까지 본원에 내원 했던 성인 중간부 쇄골 골절 환자 194명 중 완전 전위를 보인 78례를 조사하여 이중 63례를 대상으로 분석하여 후 향적 연구를 시행하였다. 4개월 추시 후에도 방사성 사진상 유합 소견이 관찰되지 않으면 불유합의 유의성을 알아보고자 하였다. 결 과 : 63례 중 불유합 및 지연 유합을 소견을 보인 환자는 15(23%)례였다. 골유합을 보인 48례의 단축은 평균 8.6㎜(2㎜-17㎜)였고, 전위는 평균 9.7㎜(2-22㎜)였다 .불유합 및 지연 유합을 보였던 15례에서 단축은 평균 14.5㎜(3㎜-37㎜), 전위는 평균 17.3㎜(4-25㎜)였다. 18㎜이상 단축이 있었던 경우 불유합의 발생과는 통계학적으로 유의성을 나타내었고(Fisher's exact test, p<0.01), Chi-square test상 16㎜ 이상의 전위를 보였던 경우 불유합 발생과 통계학적으로 의미가 있었다(p<0.01). 결 론 : 쇄골 골절 특히 중간 1/3의 골절은 비교적 보존적 치료에 결과가 좋은 것으로 되어 있으나, 전위 및 단축이 심한 경우 불유합에 대한 가능성이 높이 때문에 보다 경과 관찰시 유합의 진행이 없으면 적극적인 치료를 고려해야한다. Purpose : Because the prognosis of the mid 1.3 clavicle fracture is good, the conservative treatment with a figure of 8 bandage is the gold standard and the nonunions are rare. However, recently surgical treatment is recommended when the shortening and displacement is severe because of the high nonunion rate and the poor clinical result. This study was undertaken to evaluate that the shortening and displacement at fracture site are associated with the development of nonunion. Materials and Methods : We analysed the 194 fractures of mid 1/3 clavicle in adults which had been treated conservatively from February 1993 to January 2002 and did the retrospective study. Of these, 78 cases were originally in the middle third of the clavicle and had been completely displaced. We reviewed 63 of these cases. The shortening and displacement at the fracture site was measured on the initial roentgenogram. And the analysis of the patients' chart was done for another predisposing nonunion factors. Nonunion and delayed union are considered to be present when there has been little or no progression of clinical or radiographic healing at a minimum of 4 months after injury. Results : 15 of the 63 cases had developed nonunion.. The average 8.6㎜(2㎜-17㎜) shortening and average 9.7㎜(2-22㎜) in the union patients. The average 14.5㎜(3㎜-37㎜) shortening and average 17.3㎜(4-25㎜) in the nonunion patients. We found that initial shortening ≥18㎜(Fisher's exact test, p<0.01) and initial displacement ≥ 16㎜(Chi-square test. p<0.01)at the fracture site were significantly associated with the development of nonunion. Conclusion : The conservative treatment with figure-80-bandage is the gold standard in the clavicle middle one third fracture. However, the nonunion is commonly occurs in the cases of more of severely shortened and displaced fractures. If there are no signs of callus formation and the patient complains of pain after several week, osteosynthesis should be considered.