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Reversal of multi-drug resistance by pSUPER-shRNA-mdr1 in vivo and in vitro

World Journal of Gastroenterology 2009年第4期15卷 431-440页

作者： Guang-Dong Pan Jian-Qing yang lv-nan yan Guang-Ping Chu Qiang Liu Yi Xiao Lin Yuan Department of General Surgery People's Hospital of Liuzhou Medical Universityof Guangxi Liuzhou 545001 Guangxi Zhuang Autonomous Region China

AIM： To explore the possibility of reversing multi-drug resistance （MDR） to HepG2/mdr1 in vitro and in vivo with RNA interference （RNAi）. METHODS： HepG2/mdr1 was obtained by cloning the whole gene mdrl into HepG2 cells, shRNA targeting sequence was designed to be homologous to the P-gp encoding MDR1 mRNA consensus sequence, pSUPER- shRNA/mdrl was constructed using the enzyme- digested technique. HepG2/mdrl cells were transfected with vectors of pSUPER-shRNA/mdrl to measure their efficacy by real-time PCR for mdrl mRNA, flow cytometry （FCM） for P-gp expression, and Rhodamine efflux, MTT method for HepG2/mdrl function, respectively. In vivo, mice tumors were treated by injecting pSUPER-shRNA/mdrl in situ and into intra- abdominal cavity. Tumors were collected to create cell suspension and cryosections after chemothearpy with adiramycin and mytomycin. The cell suspension was incubated in RPMI-1640 supplemented with G418 to screen stable cells for appreciating the reversal of MDR. Cryosections were treated with immunohistochemistry technique to show the effectiveness of transfection and the expression of ***： pSUPER-shRNA/mdrl was successfully constructed, which was confirmed by sequencing. The MDR phenotype of HepG2/mdr1 was decreased significantly in vitro transfection. HepG2/mdr1 showing its MDR was reversed notably in P-gp expression （11.0% vs 98.2%, P 〈 0.01）. Real-time PCR showed that mRNA/mdrl was lower in test groups than in control groups （18.73± 1.33 vs 68.03±2.21, P 〈 0.001）. Compared with HepG2, the sensitivity of HepG2/ mdrl and HepG2/mdr1-dsRNA cells to ADM was decreased by 1.64 times and 15.6 times, respectively. The accumulation of DNR in positive groups was decreased evidently. In vivo, the p-gp expression in positive groups was significantly lower than that in control groups （65.1% vs 94.1%, P 〈 0.05）. The tumor suppressing rate in test groups was 57.8%. After chemotherapy, the growth rate in test groups was lower than that in control groups （

关键词： Hepatocellular carcinoma Multi-drugresistance RNA interference

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Salvage liver transplantation in the treatment of hepatocellular carcinoma:A Meta-analysis

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World Journal of Gastroenterology 2012年第19期18卷 2415-2422页

作者： Hong-Yu Li Yong-Gang Wei lv-nan yan Bo Li Department of Liver and Vascular Surgery Center of Liver TransplantationWest China HospitalSichuan UniversityChengdu 610041Si-chuan ProvinceChina

AIM:To evaluate survival and recurrence after salvage liver transplantation(SLT) for the treatment of hepatocellular carcinoma(HCC) compared with primary liver transplantation(PLT) using a ***:Literature on SLT versus PLT for the treatment of HCC published between 1966 and July 2011 was retrieved.A meta-analysis was conducted to estimate pooled survival and disease-free rates.A fixed or random-effect model was established to collect the ***:The differences in overall survival and disease-free survival rates at 1-year,3-year and 5-year survival rates were not statistically significant between SLT group and PLT group(P > 0.05).After stratifying the various studies by donor source and Milan criteria,we found that:(1) Living donor liver transplantation recipients had significantly higher 1-year survival rate,lower 3-year and 5-year survival rates compared with deceased-donor liver transplantation(DDLT) *** in DDLT recipients they had better 1-year and 5-year disease-free survival rate in SLT group;and(2) No difference was seen in 1-year,3-year and 5-year survival rates between two groups who beyond Milan criteria at the time of liver ***:SLT can be effectively performed for patients with recurrence or deterioration of liver function after hepatectomy for *** does not increase the perioperative mortality and has a similar long-term survival rates compared to PLT.

关键词： Salvage liver transplantation Primary livertransplantation Hepatocellular carcinoma Meta-analy-sis Survival rate

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Living donor liver transplantation using dual grafts:Ultrasonographic evaluation

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World Journal of Gastroenterology 2010年第31期16卷 3979-3983页

作者： Qiang Lu Yu-Ting Fan yan Luo Hong Wu lv-nan yan Zhe-Yu Chen Department of Sonography West China Hospital Sichuan University Chengdu 610041 Sichuan Province China Liver Transplantation Center We st China HospitalSichuan UniversityChengdu 610041Sichuan ProvinceChina

AIM: To evaluate the dual-graft living donor liver transplantation (LDLT) with ultrasonography, with special emphasis on the postoperative complications. METHODS: From January 2002 to August 2007, 110 adult-to-adult LDLTs were performed in West China Hos- pital of Sichuan University. Among them, dual-graft implantations were performed in six patients. Sonographic findings of the patients were retrospectively reviewed. RESULTS: All the six recipients survived the dual-graft adult-to-adult LDLT surgery. All had pleural effusion. Four patients had episodes of postoperative abdominal complications, including fluid collection between the grafts in three patients, intrahepatic biliary dilatation in two, hepatofugal portal flow of the left lobe in two, and atrophy of the left lobe in one. CONCLUSION: Although dual-graft LDLT takes more efforts and is technically complicated, it is safely feasible. Postoperative sonographic monitoring of the recipient is important.

关键词： Liver transplantation Dual grafts Complications Ultrasonography

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p53 codon 72 polymorphism and liver cancer susceptibility: A meta-analysis of epidemiologic studies

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World Journal of Gastroenterology 2011年第9期17卷 1211-1218页

作者： Xi Chen Fei Liu Bo Li Yong-Gang Wei lv-nan yan Tian-Fu Wen Department of Liver and Vascular Surgery West China Hospital Sichuan University 37 Guo Xue Road Chengdu610041 Sichuan Province China

AIM:To evaluate the association between p53 codon 72 polymorphism and liver cancer risk by means of meta-analysis. METHODS:Two investigators independently searched the Medline,Embase and Chinese Biomedicine *** odds ratios and 95%CI for p53 codon 72 polymorphism and liver cancer were calculated in fixedeffects model(Mantel-Haenszel method)and randomeffects model(DerSimonian and Laird method)when appropriate. RESULTS:This meta-analysis included 1115 liver cancer cases and 1778 *** combined results based on all studies showed that there was a statistically significant link between Pro/Pro genotype and liver cancer,but not between Arg/Arg or Pro/Arg genotype and liver *** stratifying for race,similar results were obtained,*** with liver cancer had a significantly higher frequency of Pro/Pro genotype than non-cancer patients among *** stratifying thevarious studies by control source,gender,family history of liver cancer and chronic hepatitis virus infection,we found that(1)patients among hospital-based studies had a significantly higher frequency of Pro/Pro and a significantly lower frequency of Arg/Arg genotype than individuals without cancer;(2)female patients with liver cancer had a significantly lower frequency of Arg/Arg and a higher frequency of Pro/Arg+Pro/Pro genotypes than female individuals without cancer;(3)subgroup analyses for family history of liver cancer did not reveal any significant association between p53 codon 72 polymorphism and liver cancer development;and(4) patients with negative hepatitis virus infection had a significantly higher frequency of Pro/Pro and a significantly lower frequency of Arg/Arg genotype than individuals without cancer. CONCLUSION:This meta-analysis suggests that the p53 codon 72 polymorphism may be associated with liver cancer among Asians.

关键词： Liver cancer p53 codon 72 Gene polymorphism Meta-analysis

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Interleukin-10-1082G/A polymorphism and acute liver graft rejection:A meta-analysis

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World Journal of Gastroenterology 2012年第8期18卷 847-854页

作者： Fei Liu Bo Li Wen-Tao Wang Yong-Gang Wei lv-nan yan Tian-Fu Wen Ming-Qing Xu lia-Yin yang Department of Liver and Vascular Surgery West China HospitalSichuan UniversityChengdu 610041Sichuan ProvinceChina

AIM: To investigate the association between interleukin (IL)-10-1082 (G/A) promoter polymorphism and acute rejection (AR) in liver transplant (LT) recipients. METHODS: Two investigators independently searched the Medline, Embase, China National Knowledge Infrastructure, and Chinese Biomedicine Databases. Summary odds ratios (ORs) and 95% CIs for IL-10-1082 G/A polymorphism and AR were calculated in a fixed and a random-effects model as appropriate. RESULTS: This meta-analysis included seven casecontrol studies, which comprised 652 cases of LT recipients in which 241 cases developed AR and 411 cases did not develop AR. Overall, the variant A allele was not associated with AR risk when compared with the wild-type G allele (OR = 0.94, 95% CI: 0.64-1.39). Moreover, similar results were observed when the AA genotype was compared with the AG/GG genotype (OR = 1.05, 95% CI: 0.55-2.02). When stratifying for eth-nicity, no significant association was observed among either Caucasians or Asians. Because only one study was performed in Asian patients, the result of subgroup analysis by ethnicity would not be reliable for Asians. Limiting the analysis to the studies with controls in the Hardy-Weinberg equilibrium, the results were persistent and robust. No publication bias was found in the present study. CONCLUSION: This meta-analysis suggests that IL-10-1082 G/A polymorphism may be not associated with AR risk in LT recipients among Caucasians.

关键词： Liver transplantation Acute rejection In-terleukin-10 Gene polymorphism Meta-analysis

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Adult-to-adult living donor liver transplantation for acute liver failure in China

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World Journal of Gastroenterology 2012年第48期18卷 7234-7241页

作者： Ding Yuan Fei Liu Yong-Gang Wei Bo Li lv-nan yan Tian-Fu Wen Ji-Chun Zhao Yong Zeng Ke-Fei Chen Department of Liver and Vascular Surgery and Liver Transplantation Center of West China Hospital Sichuan University Department of Hepatopancreatobiliary Surgery of West China Hospital Sichuan University

AIM:To investigate the long-term outcome of recipients and donors of adult-to-adult living-donor liver transplantation(AALDLT) for acute liver failure(ALF).METHODS:Between January 2005 and March 2010,170 living donor liver transplantations were performed at West China Hospital of Sichuan *** living liver donor was voluntary and provided informed *** ALF patients underwent AALDLT for rapid deterioration of liver *** was defined based on the criteria of the American Association for the Study of Liver Diseases,including evidence of coagulation abnormality [international normalized ratio(INR) ≥ 1.5] and degree of mental alteration without pre-ex-isting cirrhosis and with an illness of < 26 wk *** reviewed the clinical indications,operative procedure and prognosis of AALDTL performed on patients with ALF and corresponding living *** potential factors of recipient with ALF and corresponding donor outcome were respectively investigated using multivariate *** rates after operation were analyzed using the Kaplan-Meier *** operator characteristic(ROC) curve analysis was undertaken to identify the threshold of potential risk ***:The causes of ALF were hepatitis B(n = 18),drug-induced(n = 1) and indeterminate(n = 1).The score of the model for end-stage liver disease was 37.1 ± 8.6,and the waiting duration of recipients was 5 ± 4 *** graft types included right lobe(n = 17) and dual graft(n = 3).The mean graft weight was 623.3 ± 111.3 g,which corresponded to graft-torecipient weight ratio of 0.95% ± 0.14%.The segment Ⅴor Ⅷ hepatic vein was reconstructed in 11 right-lobe *** 1-year and 3-year recipient's survival and graft survival rates were 65%(13 of 20).Postoperative results of total bilirubin,INR and creatinine showed obvious improvements in the survived ***,the creatinine level of the deaths was increased postoperatively and became more aggravated compared with the level

关键词： Acute liver failure Adult-to-adult liver donor liver transplantation Recipient Donor Risk factors

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Kupffer细胞与肝脏缺血再灌注损害的研究进展

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世界华人消化杂志 2008年第24期16卷 2741-2745页

作者：刘作金严律南四川大学华西医院普外二科四川省成都市610041

全身80%-90%的巨噬细胞为位于肝血窦内的Kupffer细胞(KC).KC激活后将诱发TNF-α、前列腺素、一氧化氮及氧自由基等各种炎症细胞因子的大量形成,除导致自身功能形态发生改变外,还直接影响邻近的肝细胞,血管内皮细胞以及位于血窦腔内的中... 详细信息

全身80%-90%的巨噬细胞为位于肝血窦内的Kupffer细胞(KC).KC激活后将诱发TNF-α、前列腺素、一氧化氮及氧自由基等各种炎症细胞因子的大量形成,除导致自身功能形态发生改变外,还直接影响邻近的肝细胞,血管内皮细胞以及位于血窦腔内的中性粒细胞等多种细胞,进而启动热缺血或冷缺血后肝脏的缺血再灌注损害(ischemia reperfusion injury,IRI).本文拟就KC与肝脏IRI的关系进行综述,以期为IRI的防治寻求到适宜的治疗靶点及途径.

关键词： Kupffer细胞肝脏缺血-再灌注损害

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Tacrolimus dosage requirements in living donor liver transplant recipients with small-for-size grafts

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World Journal of Gastroenterology 2009年第31期15卷 3931-3936页

作者： Fei Liu Ya Li Xiang Lan Yong-Gang Wei Bo Li lv-nan yan Tian-Fu Wen Ji-Chun Zhao Ming-Qing Xu Wen-Tao Wang Jia-Yin yang Department of Liver and-Vascular Surgery Center of Liver Transplantation West China Hospital Sichuan University Chengdu 610041 Sichuan Province China West China School of Clinical Medicine Sichuan University Chengdu 610041 Sichuan Province China

AIM： To investigate the tacrolimus dosage requirements and blood concentrations in adult-to-adult right lobe living donor liver transplantation （AALDLT） recipients with small-for-size （SFS） grafts. METHODS： During January 2007 and October 2008, a total of 54 cases of AALDLT with an observation period of 6 mo were enrolled in this study. The 54 patients were divided into two groups according to graftrecipient body weight ratio （GRBW）： SFS grafts group （Group S, GRBW 〈 0.8%, n = 8） and non-SFS grafts group （Group N, GRBW ≥ 0.8%, n = 46）. Tacrolimus 12-hour blood levels and doses were recorded during weeks 1, 2, 3 and 4 and months 2, 3, 4, 5 and 6 in group S and group N. Meanwhile, acute rejection rates, liver and renal function test results, and the number of potentially interacting medications were determined at each interval in the two groups. A comparison of tacrolimus dosage requirements and blood levels were made weekly in the first month post-surgery, and monthly from months 2 to 6. RESULTS： There were no differences in the demo-graphic characteristics, acute rejection rates, liver and renal function test results, or the number of potentially interacting medications administered between the two groups. The tacrolimus dosage requirements in group S were significantly lower than group N at 2 wk （2.8 ± 0.4 mg/d vs 3.6 ± 0.7 mg/d, P = 0.006）, 3 wk （2.9± 0.7 mg/d vs 3.9±0.8 rag/d, P = 0.008）, 4 wk （2.9 ± 0.8 mg/d vs 3.9 ± 1.0 mg/d, P = 0.023） and 2 mo （2.8 ±0.7 mg/d vs 3.8±1.1 mg/d, P = 0.033）. Tacrolimus 12-h trough concentrations were similar between the two groups at all times except for 2 wk post-transplantation, when the concentrations were significantly greater in group S recipients than in group N recipients （11.3 ± 4.8 ng/mL vs 7.0 ± 3.8 ng/mL, P = 0.026）. CONCLUSION： SFS grafts recipients have significantly decreased tacrolimus dosage requirements compared with non-SFS grafts recipients in AALDLT during the first 2 mo post-surgery.

关键词： Blood concentration Liver transplantation Living donor Small-for-size graft Tacrolimus

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UPLC法同时测定三黄睛视明丸中9个成分含量

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药物分析杂志 2018年第10期38卷 1677-1682页

作者：吕燕男夏忠庭何毅丁黎中国药科大学药物分析教研室南京210009 天士力控股集团有限公司研究院现代中药开发中心天津300410 天士力医药集团股份有限公司创新中药关键技术国家重点实验室天津300410

目的:建立超高效液相色谱法同时测定三黄睛视明丸中香蒲新苷、毛蕊异黄酮苷、异鼠李素-3-O-新橙皮苷、芒柄花苷、三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd 9个成分的含量。方法:采用Waters Acquity UPLC?BEH C18... 详细信息

目的:建立超高效液相色谱法同时测定三黄睛视明丸中香蒲新苷、毛蕊异黄酮苷、异鼠李素-3-O-新橙皮苷、芒柄花苷、三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd 9个成分的含量。方法:采用Waters Acquity UPLC?BEH C18色谱柱(2.1 mm×100 mm,1.7μm),柱温30℃,流动相为水(A)-乙腈(B),梯度洗脱,流速0.3 mL·min^(-1),检测波长203 nm。结果:香蒲新苷、毛蕊异黄酮苷、异鼠李素-3-O-新橙皮苷、芒柄花苷、三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的线性范围分别为1.92～36.94 ng(r=0.999 8)、1.90～36.63 ng(r=0.999 9)、1.88～36.20 ng(r=0.999 6)、0.99～19.09 ng(r=0.999 2)、40.04～770.05 ng(r=0.999 8)、107.78～2 072.68 ng(r=0.999 8)、31.74～610.34 ng(r=0.999 1)、81.33～1 564.08 ng(r=0.999 9)、19.711～379.06 ng(r=0.999 9);平均回收率(n=6)均在91%～105%之间,RSD均小于3.0%。测定的3批样品中上述9个成分含量范围分别为0.261～0.294、0.196～0.209、0.189～0.193、0.101～0.111、2.728～2.773、12.051～12.196、3.235～3.313、8.317～8.464、2.131～2.229 mg·g-1。结论:所建立的多成分含量测定方法可用于三黄睛视明丸质量控制。

关键词：三黄睛视明丸香蒲新苷毛蕊异黄酮苷异鼠李素-3-O-新橙皮苷芒柄花苷三七皂苷人参皂苷多成分含量测定超高效液相色谱

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香橘乳癖宁胶囊UPLC特征指纹图谱研究

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中草药 2018年第2期49卷 346-352页

作者：李响白雪何毅吕燕男缪兴龙周桂荣李萍中国药科大学天然药物活性组分与药效国家重点实验室南京210009 天士力控股集团有限公司研究院现代中药开发中心天津300410 天士力医药集团股份有限公司创新中药关键技术国家重点实验室天津300410

目的建立香橘乳癖宁胶囊(XRC)的UPLC指纹图谱,利用系统指纹定量法鉴定各批次制剂质量,并采用电喷雾离子源-四极杆-飞行时间质谱(ESI-Q-TOF/MSE)对其化学成分进行定性分析。方法采用Acquty UPLC BEH C18(100mm×2.1 mm,1.7μm)色谱柱,以0... 详细信息

目的建立香橘乳癖宁胶囊(XRC)的UPLC指纹图谱,利用系统指纹定量法鉴定各批次制剂质量,并采用电喷雾离子源-四极杆-飞行时间质谱(ESI-Q-TOF/MSE)对其化学成分进行定性分析。方法采用Acquty UPLC BEH C18(100mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,体积流量为0.25 m L/min,柱温为28℃,检测波长为270 nm。以系统指纹定量法的宏定性相似度(Sm)、宏定量相似度(Pm)为评价指标,对10批制剂的整体质量进行评价。并利用UPLC-ESI-Q-TOF/MSE正、负2种离子模式扫描对共有峰成分进行定性分析。结果建立10批XRC的UPLC对照指纹图谱,标定33个共有峰,指认其中28个成分,并分别归属药材来源。系统指纹定量法评价结果表明,9批制剂质量在5级以上。结论建立有效、可靠的XRC制剂质量评价方法,对其质量控制方法进行提升,并为物质基础的进一步研究奠定基础。

关键词：香橘乳癖宁胶囊电喷雾离子源-四极杆-飞行时间质谱系统指纹定量法指纹图谱丹酚酸B

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