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(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.

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.

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.

전기영동에 의한 동양란(Cymbidium spp.)의 단백질 유형비교

윤경은,홍성숙,이남희 서울여자대학교 자연과학연구소 1991 자연과학연구논문집 Vol.2 No.-

본실험은 전기영동을 이용하여 단백질과 동위효소의 유형을 비교 분석함으로 종과 품종을 구분할 수 있는가의 가능성을 조사하고 전기영동에 적합한 난의 부위와 효소의 종류를 선정코자 실시하였다. 1. 한국춘란은 부위별로 꽃봉오리, 뿌리끝, 어린잎, 성엽을 6% polyacrylamide slab gel로 단백질을 비교한 결과 부위별로 유형을 달리하였으나 band의 분리가 명확하지 않았다. 2. 10% SDS polyacrylamide slab gel에서는 band의 분리가 명확하였고 영동재료로는 어느 때나 쉽게 채취가 가능한 잎이나 뿌리를 사용하는 것이 편리하나 뿌리는 phenol 함량은 적으나 단백질 함량이 극히 낮으므로 잎을 시료로 쓰는 것이 양호한 편이었다. 3. 잎을 시료로 쓰는 경우는 엽록소, phenol의 제거가 필요하나 배양중인 rhizome을 사용하면 이러한 난점이 없이 단백질 유형이 뚜렷이 나타나, 춘란(Cymbidium goeingii), 한란(C, kanran), 죽백란(C, lancifolium)을 구분할 수 있었다. 4. Rhyzome의 esterase, peroxidase, acid phosphatase, glucose phosphate isomerase의 동위효소를 비교한 결과 단백질 유형으로는 분류가 어려웠던 춘란(Cymbidium goeringii)과 춘란복륜(Cymbidium goeringii:variated leaves)도 구분할 수 있었다. Peroxidase는 band는 여러가지로 나타났으나 esterase와 같이 band pattern이 명확지 못하였고 acid phosphatase와 glucose phosphate isomerase는 한개씩의 band만이 나타나 분류에는 적합하지 않은 효소로 생각되어졌다. The buffer soluble proteins were extracted from flower, root, young leaves, old leaves, and cultured rhizome. Different patterns of general protein were observed among the tested organs. Among the samples rhizome were the most suitable for electrophoresis to differentiate cultivars. Band patterns of general protein, esterase, peroxidase showed typical band patterns among Cymbidium goeringii, Cymbidium kanran, and Cymbidium lancifolium which have similar appearance of vegetative leaves. The isozymes of esterase, peroxidase, acid phosphatase, and glucose phosphate isomerase in cultured rhizome were analized. The well-resolved zones of esterase activity were detected. The differentiation of Cymbidium goeringii and Cymbidium goeringii : variated leaves which was hard to differentiated by general protein patterns, could be done by esterase isozyme band pattern.

고출력 펄스 레이저로 증착된 유기필름

홍진수,채희백,안성기,박해윤 순천향대학교 기초과학연구소 2001 순천향자연과학연구 논문집 Vol.7 No.2

유기발광 응용에 성공적으로 사용되어 새로운 물질을 개발할 때 기준으로 사용되는 Alq_3와 TPD필름을 기존의 증착법(열증착법과 스핀코터 기술)을 대체할 수 있는 고출력펄스레이저로 합성하였다. 펠릿 타겟은Q-스위칭 Nd:YAG 레이저의 532 nm와 355 nm를 사용하여 2&10^-6 Torr압력 하에서 용발시킨다. 슬라이드 유리 위에 증착된 필름은 유기발광 디바이스에 사용될 가능성이 있는지 알기 위해 다음과 같은 조사를 하였다: PL,AFM, 흡수 스펙트라, XRD. 355 nm 레이저로 유기필름을 증착하면 532 nm 경우의 필름보다 결합이 적고, 특히 열증착 필름보다 스트레스를 덜 받는다는 사실을 알 수 있다. 또한 타겟의 구조가 Alq_3 다를지라도 레이저 증착법으로 합성된 Alq_3 필름은 열증착과 동일한 구조를 갖는다는 사실을 알 수 있다. 어느 레이저를 사용하건 증착된 필름은 결정구조를 확인할 수 없을 정도로 비정질을 이룬다는 사실을 알 수 있다.

有機農業의 實態와 發展에 관한 事例 硏究 : 忠南 洪城郡 洪東面 有機農家를 中心으로

尹英相,洪性贊,金成敏,具漢謨,申東一,金昌護,姜熙慶 公州大學校 資源科學硏究所 1997 資源科學硏究論文集 Vol.5 No.-

This study was carried out to improve the organic farming system about crop cultivation, compost production and application, plant diseases & pest control, weed control and marketing system of organic farming products to the case study in Chungnam Hongdong area. Agricultural technologies concerning pest & diseases control, weed control and compost production are not established due to lacks of systemized researches by the agricultural scientist, and agricultural machines profitable to this works had not developed yet. In cropping system, reasonable crop rotation had not conducted. The related cultivation of vegetables on the same ground for a long time cause a various plant disease easily. So crop rotation is a reasonable method about soil nutrient utility and soil disease control. It is a good method that introduce the barley and wheat in this field crop system. Those crop have a long root, so can use the other nutrient against the vegetable. In organic farming products, the safety and quality of products by the National Agricultural Products Inspection Center had not inspected except rice, lettus and apple. Inspection is needed for the development of organic farming and mutual confidence between producer and consumer, because the mutual confidence is a major thing to sustain the organic farming. In products marketing, it is more need for the information of organic farming products. Nowaday most products are marketed in Seoul region, but it is not a good method. To search the consumer in near region is a good method for decrease the cost of conveyance and maintain the quality of products. The varieties of distribution structure of organic farming products are required. The products are sold to department store in large city and the prices are about 30% higher than normal products. But when those are supplied in near region, the prices of those are similar to normal products. Direct transactions in near regions is a good method for decrease the cost of conveyance and strength the mutual confidence between producer and consumer.

근관충전방법에 따른 내흡수 치근의 근관충전 효율성 비교

홍찬의,신동훈,하상윤 大韓齒科保存學會 1995 Restorative Dentistry & Endodontics Vol.20 No.2

당뇨병환자에서 게이트심장혈액풀신티그라피를 이용한 심기능 평가

윤상임,송치운,이진홍,안미애,성기양,송민호,이강욱,신영태,김영건,노흥규 충남대학교 의과대학 지역사회의학연구소 1993 충남의대잡지 Vol.20 No.2

Major cardiovascular complications of diabetes are coronary atherosclerosis, diabetic dilated cardiomyopathy, autonomic neuropathy and those are major causes of morbidity and mortality in diabetic patients. Gated blood pool heart scan is noninvasive and useful method for evaluation of functional status of heart in diabetics. We evaluated 52 patients with diabetes and divided 3 groups. Group 1 were 11 patients without proteinuria or with proteinuria less than 550mng during 24 hours. Group 2 were 9 patients with proteinuria more than 550mg during 24 hours and group 3 were 32 patients with endstage renal diasese due to diabetes. We performed 99mTc-HSA cardiac gated blood pool scan and used left ventricular ejection fraction(LVEF), peak ejection rate(PEF) to indices of LV systolic function and peak filling rate(PER) to index of LV diastolic function. The results were follows : 1) LVEF, PER were significantly lower in diabetics with ESRD than diabetics without ESRD, but there were no significant difference between normal controls and diabetics without ESRD 2) PFR was significantly lower in diabetics than normal controls, but there were no significant differences in diabetics with or without nephropathy. 3) There were negative correlation between PER, PFR and duration of diabetes. On the basis of results, PFR is a LV functional index of GBPS which can disclose early change of LV dysfunction in patients with diabetes.