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Effects of multidrug resistance, antisense RNA on the chemosensitivity of hepatocellular carcinoma cells

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hepatobiliary & Pancreatic Diseases International 2006年第4期5卷 552-559页

作者： Bo Li, Tian Ye, Lei Zhao, De-Hua Li, Xing-Hua Gou, Lan-Ying Zhao, Lei Han, Lin Chen, Lu-Nan Yan and Jian-Ping Gong department of hepatobiliary surgery, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400010, China department of Traditional Chinese surgery, Second Affiliated Hospital, Luzhou Medical College, Luzhou 646000, China Genetic Engineering Laboratory, Chengdu Di’ao Group Co., Ltd., Chengdu 610041, China department of General surgery, West China Hospital, Sichuan University, Chengdu 610041, China department of hepatobiliary surgery Second Affiliated Hospital Chongqing University of Medical Sciences Chongqing 400010 China.

BACKGROUND: Multidrug resistance is a major obstacle in cancer chemotherapy. We examined whether the antisense RNA of multidrug resistance gene 1 (mdr1) could reverse multidrug resistance in the human hepatocellular carcinoma (HCC) cell line SMMC7721/ADM. METHODS: The recombinant adenoviruses pAdEasy- GFP-ASmdr1 product was produced by the adenoviral vector AdEasy system, which can express antisense RNA against the mdr1 gene. Following that, the recombinant adenovirus was transfected into the P-glycoprotein- producing multidrug resistance cell line, SMMC7721/ADM human HCC cells resistant to adriamycin (ADM) and daunorubicin (DNR). In order to investigate the reversal of multidrug resistance phenotype, we measured the expression of mdr1 mRNA by RT-PCR and the production of P-glycoprotein by flow cytometry. The sensitivities for ADM and DNR SMMC7721/ADM cells were examined by [3-(4, 5-dimethylthi-azol-2-yl)-2,5 diphenyl-terazolium bromide] (MTT) analysis. RESULTS: The low-level expression of mdr1 mRNA and P-glycoprotein production were observed in parental sensitive cells SMMC/7721 in addition to the overexpressionof mdr1 mRNA and P-glycoprotein in SMMC7721/ADM cells. The transfection of antisense-RNA into SMMC7721/ ADM cells resulted in decreases of mdr1 mRNA and P-glycoprotein, but increase of drug sensitivities. The sensitivities of transfected SMMC7721/ADM cells to ADM and DNR in IC50 reduced by 31.25% and 62.96% respectively. CONCLUSIONS: Mdr1 antisense RNA can increase the sensitivities of SMMC7721/ADM cells to anticancer drug by decreasing the expression of the mdr1 gene and inhibiting P-glycoprotein expression. This strategy may be applicable to cancer patients with P-glycoportein mediated multidrug resistance.

关键词： antisense nucleic acid multiple drug resistance P-glycoprotein

Association of polymorphisms in non-classic MHC genes with susceptibility to autoimmune hepatitis

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hepatobiliary & Pancreatic Diseases International 2012年第2期11卷 125-131页

作者： Jie Tang,Cheng Zhou,Zhi-Jun Zhang and Shu-Sen Zheng Hangzhou,China Key Laboratory of Combined Multi-organ Trans- plantation, Ministry of Public Health, department of hepatobiliary and Pancreatic surgery , and State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China [a] Key Laboratory of Combined Multi-organ Transplantation Ministry of Public Health Department of Hepatobiliary and Pancreatic Surgery Hangzhou China [b] State Key Laboratory for Diagnosis and Treatment of Infectious Disease First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

BACKGROUND:Autoimmune hepatitis is a chronic,generally progressive inflammatory disorder of the liver,of which the cause is *** was demonstrated that genetic factors are involved in its *** studies showed that human leukocyte antigen in the major histocompatibility complex(MHC) is associated with susceptibility to autoimmune *** genome scanning studies suggest that genes outside the MHC also play a critical role in autoimmune *** article focuses on our current understanding of the polymorphisms of these genes and their roles in the pathogenesis of autoimmune *** SOURCES:Studies were identified by searching MEDLINE and PubMed for articles using the keywords autoimmune hepatitis,polymorphism,CTLA-4,Fas,TNF-α TGF-β1,TBX21 and VDR up to May *** papers were identified by a manual search of the references from key ***:According to the case-control studies on genetic polymorphisms,at least six genes(CTLA-4,Fas,TNF-α TGF-β1,TBX21 and VDR) are involved in autoimmune hepatitis besides *** far,there has been no agreement about gene susceptibility and the actual clinical significance of these genes is still ***:Studies on gene polymorphisms outside the MHC and knowledge of genetic predispositions for autoimmune hepatitis may not only elucidate pathogenic mechanisms,but also provide new targets for therapy in the future.

关键词： autoimmune hepatitis polymorphism gene expression

The U937 cell line induced to express CD14 protein by 1,25-dihydroxyvitamin D3 and be sensitive to endotoxin stimulation

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hepatobiliary & Pancreatic Diseases International 2005年第1期4卷 84-89页

作者： Hai-Zhong Liu, Jian-Ping Gong, Chuan-Xin Wu, Yong Peng, Xu-Hong Li and Hai-Bo You Chongqing, China department of hepatobiliary surgery, Second College of Clinical Medicine & Hospital of Chongqing University of Medical Sciences, Chongqing 400010, China department of General surgery Second College of Clinical Medicine & Second Affiliated Hospital of Chongqing University of Medical Sciences Chongqing 400010 China. haizhong97@***

BACKGROUND: CD14 was first described as a differentia- tion antigen on the surface of myeloid lineage cells. It acts as a glycosylphosphatidylinositol ( GPI)-anchored receptor for the complex of lipopolysaccharide (LPS) and plays a key role in the activation of LPS-induced monocytes. The purpose of this study was to observe the expression of CD14 protein and its gene in the human U937 promonocytic cell line when these cells were exposed to 1,25-dihydroxyvita- min D3 ( VitD3 ) and investigate their sensitivity to endo- toxin stimulation. METHODS: U937 cells were exposed to (0.1 μmol) VitD3 for 24 hours and were induced to express the CD14 mRNA gene and CD14 protein, then their responses were observed when they were stimulated with different concentrations of LPS for different time. RESULTS: The U937 cells induced by VitD3 were found to stably express CD14 mRNA and CD14 protein. And CD14 protein enhanced the sensitivity of U937/CD14 cells to li- popolysaccharide ( LPS ) stimulation. NF-ΚB in U937/ CD14 cells can be activated with low concentration of LPS (1 ng/ml-10 ng/ml), the TNF-α mRNA gene was in- duced , and then TNF-α was produced and released into the supernatant of culture. CONCLUSION: VitD3 can induce U937 cell to express the CD14 gene and CD14 protein and enhance the response of this type of cells to LPS stimulation.

关键词： CD14 lipopolysaccharide U937 cell line VitD3 endotoxin stimulation

Relation of abnormal gallbladder arterioles to gallbladder emptying in patients with gallstone and diabetes mellitus

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hepatobiliary & Pancreatic Diseases International 2004年第2期3卷 275-278页

作者： Xiong Ding, Jian-Ping Gong, Chang-You Lu, Chang-An Liu, Yu-Jun Shi and Gen-Wu Gao Chongqing, China department of hepatobiliary surgery, Second Clinical College, Chongqing University of Medical Science, Chongqing 400010, China department of hepatobiliary surgery Second Clinical College Chongqing University of Medical Science Chongqing 400010 China. dingpanda@***

BACKGROUND: Diabetes mellitus is thought to be related to gallstone formation in emptying the gallbladder. Diabe- tes mellitus may lead to many changes in microarterioles and micronerves; the aim of this study was to investigate the abnormality of arterioles in the gallbladder and its rela- tion to gallbladder hypomotility in patients with gallstone and diabetes mellitus. METHODS: Thirty patients with simple gallstones and 30 patients with gallstones and diabetes mellitus were analy- zed , and their gallbladder emptying function was measured with B ultrasound before operation. After operation, the arterioles of the gallbladder rinsed with periodic acid-schiff (PAS) reagent in photos were used for analysis of the tu- blar area and stereo system with the Beihang CM-2000B biological and medical photo system. RESULTS: In patients with gallstones and diabetes mellitus, the gallbladder emptying function was significantly im- paired, the area ratio of the arteriole wall to whole arte- rioles in cross section was significantly higher than that in patients with simple gallstones (0.81 ±0.09 vs. 0.58±0.15, P 0.05). CONCLUSION: In patients with diabetes mellitus, the se- dimentation of PAS positive material in the wall of arte- rioles leads to the stenosis of arterioles. It is probably con- tributive to hypomotility of the gallbladder.

关键词： gallstone diabetes mellitus arterioles gallbladder motility

Strong additive effect of calcitriol and cyclosporine A on lymphocyte proliferation in vitro and rat liver allotransplantations in vivo

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Chinese Medical Journal 2006年第24期119卷 2090-2095页

作者： ZHANG Ai-bin and ZHENG Shu-sen department of hepatobiliary surgery,First Affiliated Hospital of Medical College,Zhejiang University Key Lab of Combined Multi-Organ Transplantation,Ministry of Health,Hangzhou 310003,China department of hepatobiliary surgery First Affiliated Hospital of Medical College Zhejiang University

Background Vitamin D3 and its metabolites have been found to exert immunosuppressive effects both in vivo and in vitro. We investigated the synergistic effect of calcitriol and cyclosporine A （CsA） on lymphocyte proliferation in vitro and graft rejection following rat liver allotransplantations in vivo. Methods Alloantigen driven, human peripheral mononuclear cells＇ proliferation and cytokine production capacity were tested in the presence or absence of various concentrations of calcitriol or CsA. In vivo, liver allografts were transplanted in a high responder strain combination （SD to Wistar） rats and combination of subtherapeutical dose of CsA and calcitriol was administered in recipients, whereas the control recipients received single or no immunosuppressant. Proliferation of splenocyte from recipient was tested with mixed lymphocyte reaction. Serum intedeukin-2 （IL-2） and interferon gamma （IFN-γ） concentrations were measured with enzyme linked immunosorbent assay. Results Combined medication of 10^-9 mol/L calcitriol and 100 ng/ml CsA inhibited human peripheral mononuclear cells＇ proliferation to alloantigen and the production of IL-2 and IFN-γ but promoted that of IL-4 and IL-10. Similarly, combination of 250 ng · kg^-1·-d^-1 calcitriol and 1.0 mg · kg^-1·-d^-1 CsA showed an additive effect in liver transplant model. It restrained splenocyte proliferation to alloantigen from donor and significantly reduced serum concentration of IL-2 and IFN-γ in recipients. Consequently, aUograft rejection in combined medication group was minor （median William＇s grade was 1.0 vs 3.0 in combined medication group and in the control group, P〈0.05） and the recipients＇ survival was evidently prolonged [（93.7±5.8） days vs （12.6±1.4） days in combined medication group and in the control group, P〈0.01 ]. Conclusion A combination of calcitriol and CsA has an additive effect on limiting lymphocyte proliferation and prolonging liver graft survival. With its additional immunomodulating pro

关键词： liver transplantation vitamin D cyclosporine cytokine lymphocyte graft rejection

Procedures for congenital choledochal cysts and curative effect analysis in adults

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hepatobiliary & Pancreatic Diseases International 2002年第3期1卷 442-445页

作者： Kai-Shan Tao Yong-Gang Lu Ting Wang Ke-Feng Dou the department of hepatobiliary surgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, China department of hepatobiliary surgery Xijing Hospital Fourth Military Medical University Xi'an 710032 China. taokaishan@***

Objective: To evaluate the procedures and timing of operation as well as long-term postoperative effect of congenital choledochal cysts (CCC) in adults. Methods: The procedures and timing of operation, effective rate, re-operation rate and incidence of car- cinoma after operation for 70 adult patients with CCC from January 1980 to June 1999 were analyzed retrospectively. Results: The re-operation rate of external drainage was 86% (6/7). The effective rate of internal drain- age was significantly lower than that of cyst resection (3/10 vs 45/49, X^2=20. 94, Psurgery was higher than that of selective operation (8/10 vs 6/56, X^2=24. 37, P<0.001). Conclusions: External drainage should be the first- aid measure and the therapy of choice on emergency basis. Internal drainage should never be attempted. Cyst resection with Roux-en-Y hepaticojejunostomy is recommended as the treatment of choice in selective operation.

关键词： choledochal cyst drainage surgical procedure

Effects of 5-hydroxytamine and its antagonists on hepatic stellate cells

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hepatobiliary & Pancreatic Diseases International 2006年第1期5卷 96-100页

作者： Tao Li, Shan-Geng Weng, Xi-Sheng Leng, Ji-Run Peng, Yu-Hua Wei, Dong-Cheng Mou and Wan-Xiang Wang department of hepatobiliary surgery, Peking University People’s Hospital, Beijing 100044, China department of hepatobiliary surgery Peking University People's Hospital Beijing 100044 China.

BACKGROUND: Hepatic stellate cell (HSC) plays a key role in hepatic fibrosis. This study was undertaken to investigate the expression of 5-hydroxytamine receptors in HSC and the effect of 5-hydroxytamine on biological characteristics of HSC. METHODS: Liver ex vivo perfusion of collagenase and density gradient centrifugation were used to isolate HSCs. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of 5-hydroxytamine receptor subtypes 1A, 2A, 2B and 3. Western blot hybridization was used to elucidate the effect of 5-hydroxytamine and its 2A receptor antagonist ketanserin and 3 receptor antagonist ondanosetron on the expression of transforming growth factor-β_1(TGF-β_1) and Smad4 in HSC. RESULTS: HSC expressed 5-hydroxytamine receptor subtypes 1A, 2A and 2B. 5-hydroxytamine significantly increased the expression of TGF-β_1, and Smad4 in HSC (P<0.05). This action can be antagonized by ketanserin, not by ondanosetron. CONCLUSIONS: HSC expresses 5-hydroxytamine receptors. 5-hydroxytamine could effect the biological characteristics of HSC through its receptor mediation, and may play a role in the pathogenesis of liver cirrhosis and portal hypertension.

关键词： hepatic stellate cell 5-hydroxytmine receptor transforming growth factor

Caveolin-1 is important for nitric oxide-mediated angiogenesis in fibrin gels with human umbilical vein endothelial cells

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Acta Pharmacologica Sinica 2006年第12期27卷 1567-1574页

作者： Yi-ming PAN Yong-zhong YAO Zhang-hua ZHU Xi-tai SUN Yu-dong QIU Yi-tao DING~2 department of hepatobiliary surgery,Drum Tower Hospital,Medical College of Nanjing University and hepatobiliary Institute of Nanjing University,Nanjing 210008,China departmentofhepatobiliarysurgery DrumTowerHospitalMedicalCollegeofNanjingUniversityandHepatobiliaryInstituteofNanjingUniversityNanjing210008China

Aim:The role of caveolin-1(Cav-1)in angiogenesis remains poorly *** endothelial nitric oxide(NO)synthase(eNOS),a caveolin-interacting protein,was demonstrated to play a predominant role in vascular endothelial growth fac-tor(VEGF)-induced *** purpose of our study was to examine therole of Cav-1 and the eNOS complex in NO-mediated ***:Human umbilical vein endothelial cells(HUVEC)were isolated and cultured in 3-Dfibrin gels to form capillary-like tubules by VEGF *** expression ofCav-1 and eNOS was detected by semiquantitative *** HUVEC weretreated with antisense oligonucleotides to downregulate Cav-1 *** and non-infected HUVEC were cultured in fibrin gels in the presenceor absence of VEGF(20 ng/mL)and N~G-nitro-L-arginine methyl ester(L-NAME;5 mmol/L).NO was measured using a NO assay kit and capillary-like tubules werequantified by tubule formation index using the Image J ***:RT-PCRanalysis revealed that Cav-1 levels steadily increased in a time-dependent mannerand reached their maximum after 5d of incubation,but there were no obviouschanges in eNOS mRNA expression in response to VEGF in the fibrin gel ***(20 ng/mL)can promote NO production and the formation of capillary-liketubules,and this promoting effect of VEGF was blocked by the addition ofL-NAME(5 mmol/L).When transduced HUVEC with the antisense Cav-1 oligo-nucleotides were plated in the fibrin gels,the capillary-like tubules were signifi-cantly fewer than those of the non-infected *** capillary-like tubules forma-tion and NO production of transduced HUVEC with the antisense Car-1 oligo-nucleotides cultured in fibrin gels showed no responses to the addition of VEGF(20 ng/mL)and L-NAME(5.0 mmol/L).Conclusion:NO was a critical angiogenicmediator in this ***-1 was essential for NO-mediated angiogenesis andmay be an important target of anti-angiogenesis therapy.

关键词： caveolin-1 angiogenesis nitric oxide vascular endothelial growth factor

Variations and reconstruction of the hepatic artery in liver transplantation

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hepatobiliary & Pancreatic Diseases International 2006年第2期5卷 170-172页

作者： Xiao Xu and Shu-Sen Zheng department of hepatobiliary and Pancreatic surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China

Any surgical refinement and innovation is certain to improve the long-term survival of recipients following liver transplantation. In the past de- cade, the shortage or donor organs has prompted us to use partial liver graft in split liver transplantation and living donor liver transplantation. As an anatomic problem in a successful transplantation, which actually is beyond the category of anatomy, the variations and reconstruction of hepatic vasculatures have drawn much attention of surgeons.

关键词： HAV