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The expression and antigenicity identification of recombinant rat TGF-β1 in bacteria

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Cell Research 2001年第2期11卷 95-100页

作者： gao chun fang, XIAN TAO KONG, AXEL M GRESSNER, RALF WEISKIRCHEN department of laboratory medicine, changzheng hospital, second military medical university, shanghai 200003, china Institute of Clinical Chemistry and Pathobiochemistry, Central laboratory, R 1department of laboratory medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 1department of laboratory medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 2Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH-University Hospital Pauwelsstr 30 D-52074 Aachen Germany 2Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH-University Hospital Pauwelsstr 30 D-52074 Aachen Germany

In order to study structure-function details of TGF-β1, the recombinant mature form of rat TGF-β1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-β1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. This system allowed an active and selective synthesis of recombinant TGF-β1. The molecular weight of expressed TGF-β1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF-β1 antibody. The mature recombinant rat TGF-β1 expressed in this study provides a useful tool for future detailed structural and functional studies.

关键词： TGF-β1 recombinant expression inclusion bodies purification antigenicity.

Design,synthesis and antitumor evaluation of a new series of N-substi-tuted-thiourea derivatives

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Acta Pharmacologica Sinica 2006年第9期27卷 1259-1271页

作者： Jian LI~2 Jin-zhi TAN~2 Li-li CHEN~2 Jian ZHANG~2 xu SHEN~(2,3)Chang-lin MEI~(4,6)Li-li FU~4 Li-ping LIN~5 Jian DING~5 Bing XIONG~2 Xi-shan XIONG~4 Hong LIU~(2,6)Xiao-min LUO~(2,6)Hua-liang JIANG~(2,3)2 Drug Discovery and Design Centre,State Key laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,shanghai 201203.china 3 School of Pharmacy.East china university of Science and Technology,shanghai 200237,china 4 Division of Nephrology,department of Internal medicine,changzheng hospital,second military medical university,shanghai 200003,china 5 Division of Antitumor Pharmacology,shanghai Institute of Materia Medica,shanghai 201203.china 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 3School of Pharmacy East China University of Science and Technology Shanghai 200237 China 3School of Pharmacy East China University of Science and Technology Shanghai 200237 China 4Division of Nephrology Department of Internal Medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 5Division of Antitumor Pharmacology Shanghai Institute of Materia Medica Shanghai 201203 China 5Division of Antitumor Pharmacology Shanghai Institute of Materia Medica Shanghai 201203 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 4Division of Nephrology Department of Internal Medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China 2Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Bi

Aim:To design and synthesize a novel class of protein tyrosine kinase inhibitors,featuring the N-(2-oxo-1,2-dihydroquinolin-3-yl-methyl)-thiourea ***:First,compounds 1 and 2 were identified using the virtual screeningapproach in conjunction with binding assay based on surface plasmon ***,3 regions of compounds 1 and 2 were selected for chemical *** compounds were characterized potent inhibitory activities toward thehuman lung adenocarcinoma cell line SPAC ***:Forty new compounds(1-2,3a-g,4a-w,and 5a-l)were designed,synthesized and *** com-pounds(1,3e,41,4w,5a,and 5b)were found to show promising inhibitory activityagainst the SPAC1 tumor cell *** inhibitory activity of compound 5aincreases approximately 10 times more than that of the original compound ***:This study provides a promising new template with potential antitu-mot activity.

关键词： N-substituted-thiourea derivatives antitumor SPAC1 tyrosine kinase inhibitor virtual screening

-509C＞T polymorphism in the TGF-β1 gene promoter is not associated with susceptibility to and progression of colorectal cancer in Chinese

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-509C＞T polymorphism in the TGF-β1 gene promoter is not as...

全国临床免疫检验研讨会暨第六届全国临床免疫学术会议

作者： peng qi~(1△),Can-ping Ruan~(2△),hao wang~3,Fei-guo Zhou~4,Xin-yun xu~2,Yun-peng Zhao~1, xing Gu~1,chun-fang gao~(*1) 1.department of laboratory medicine,eastern hepatobiliary hospital,second military medical university,shanghai 200438,china 2.department of General Surgery,changzheng hospital,second military medical university,shanghai 200003,china 3.department of laboratory medicine,changzheng hospital,second military medical university,shanghai 200003,china 4.department of Hepatic Surgery,eastern hepatobiliary hospital,second military medical university,shanghai 200438,china.

Objective Colorectal cancer is one of the most common cancers in the world, accounting for nearly 10%of all incident cases of *** growth factor-β1 (TGF-β1) is a pleiotropic cytokine that has been implicated in the pathogenesis of colorectal *** most studied -509C>T polymorphism of TGF-β1 gene has been associated with various kinds of *** aim was to investigate the association between this genetic variant and the risk and/or progression of colorectal cancer. Method We conducted a case-control study of 150 colorectal cancer cases and 503 healthy *** was extracted from blood specimens,and -509C>T polymorphism in the TGF-β1 gene promoter was genotyped by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).In addition,colorectal cancer tissues(n=70) were obtained from the studied cases for measurement of TGF-β1 mRNA expression *** also assessed the plasma TGF-β1 levels of cases(n=88) and healthy subjects (n=120).Results In our study group,the high TGF-β1 producer genotypes,-509TT,was not associated with an increased risk of colorectal cancer compared with other genotypes, nor with the progression of colorectal ***,we observed that colorectal cancer patients especially those with a more aggressive disease behavior were more frequent with C *** Our results suggest that TGF-β1 -509C>T polymorphism was not associated with both an increased risk and progression of colorectal cancer.

关键词： Colorectal cancer transforming growth factor-β1 single-nucleotide polymorphism case-control study individual susceptibility

Analysis of HLA alleles polymorphism in Chinese patients with primary biliary cirrhosis

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hepatobiliary & Pancreatic Diseases International 2006年第1期5卷 129-132页

作者： Hai-Ying Liu. An-Mei Deng, Ye Zhou, Ding-Kang Yao, De-xing xu and Ren-qian Zhong Clinical laboratory, General hospital of Guangzhou military Command of PLA, Guangzhou 510010, china laboratory Diagnostics, changzheng hospital, second military medical university, and Clinical Immunology Center of PLA, shanghai 200003, china and department of Gastroenterology, changzheng hospital, second military medical university, shanghai 200003, china Clinical laboratory General Hospital of Guangzhou Military Command of PLA Guangzhou 510010 China. xiangliuhaiying@***.

BACKGROUND: A familial dustering of patients with primary biliary cirrhosis (PBC) and the presence of immunological abnormalities in family members suggest a genetic component involved in the pathogenesis of PBC. The aims of this study are to investigate the frequencies of human leukocyte antigen HLA-A, -B, and -DRB1 alleles in Chinese patients with PBC by polymerase chain reaction (PCR)-based techniques, and to assess the correlation of the above-mentioned HLA with some clinical and laboratory features. METHODS: Genotyping of HLA alleles were performed in 65 well-characterized PBC patients and 431 healthy controls with sequence-specifc primers PCR amplification. RESULTS: HLA-DRB1~*07 allele detected in 19 of the 65 (29.2%) PBC patients was subtyped as DRB1~*0701, as well as in 13.9% of controls (P_C2.55, 95% CI: 1.4-4.6). An increased frequency of DRB1~*03 (18.4% vs. 7.2% in healthy controls) and a decreased frequency of DRB1~*12 (16.9% vs. 28.8%) in PBC patients were statistically significant. There was no association with HLADRB1~*08 reported. The frequencies for HLA-A, B and the other DRB1 alleles were similar between patients and healthy controls. CONCLUSIONS: The susceptibility to PBC in Chinese individuals is associated with DRB1~*0701 allele. This association differs from that in North Americans, South Americans, North Europeans and even Japanese, but it is not restricted to any particular subgroup of patients.

关键词： primary biliary cirrhosis human leukocyte antigens genetic susceptibility

Risk factors of intrahepatic cholangiocarcinoma in patients with hepatolithiasis:a case-control study

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hepatobiliary & Pancreatic Diseases International 2011年第6期10卷 626-631页

作者： Zhen-Yu Liu,Yan-Ming Zhou,Le-Hua Shi and Zheng-Feng YinAuthor Affiliations:Molecular Oncology laboratory and department of Comprehensive Treatment I,eastern hepatobiliary Surgery hospital,second military medical university,shanghai 200438,china department of Hepato-Biliary-Pancreato-Vascular Surgery,First Affiliated hospital,Xiamen university,Xiamen 361003,china Molecular Oncology laboratory Eastern Hepatobiliary Surgery Hospital Second Military Medical University Shanghai 200438 China department of Comprehensive Treatment I Eastern Hepatobiliary Surgery Hospital Second Military Medical University Shanghai 200438 China department of Hepato-Biliary-Pancreato-Vascular Surgery First Affliated Hospital Xiamen University Xiamen 361003 China

BACKGROUND:Why 3.3% to 10% of all patients with hepatolithiasis develop intrahepatic cholangiocarcinoma (ICC) remains *** carried out a hospital-based case-control study to identify risk factors for the development of ICC in patients with hepatolithiasis in ***:Eighty-seven patients with pathologically diagnosed hepatolithiasis associated with ICC and 228 with hepatolithiasis alone matched by sex,age (±2 years),hospital admittance and place of residence were interviewed during the period of *** ratios (OR) and 95% confidence intervals (CI) were calculated for each risk ***:Among the patients with hepatolithiasis associated with ICC,the mean age was 57.7 years and 61.0% were female Univariate analysis showed that the significant risk factors for ICC development in hepatolithiasis were smoking,family history of cancer,appendectomy during childhood (under age 20),and duration of symptoms >10 *** multivariate stepwise logistic regression analysis,smoking (OR=1.931,95% CI:1.000-3.731),family history of cancer (OR=5.175,95% CI:1.216-22.022),and duration of symptoms >10 years (OR=2.348 95% CI:1.394-3.952) were independent ***:Smoking,family history of cancer and duration of symptoms >10 years may be risk factors for ICC in patients with hepatolithiasis.

关键词： risk factors intrahepatic cholangiocarcinoma hepatolithiasis case-control study

Changes of serum alpha-fetoprotein and alpha-fetoprotein-L3 after hepatectomy for hepatocellular carcinoma: prognostic significance

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hepatobiliary & Pancreatic Diseases International 2012年第6期11卷 618-623页

作者： Xiao-Feng Zhang, Zheng-Feng Yin, Kui wang, Zong-qin Zhang, Hai-Hua qian,Le-Hua ShiAuthor Affiliations: The 4th department of Hepatic Surgery ,Molecular Oncology laboratory , eastern hepatobiliary Surgery hospital, second military medical university, shanghai 200438, china The 4th department of Hepatic Surgery Eastern Hepatobiliary Surgery Hospital Second Military Medical University Shanghai 200438 China Molecular Oncology laboratory Eastern Hepatobiliary Surgery Hospital Second Military Medical University Shanghai 200438 China

BACKGROUND: Alpha-fetoprotein (AFP) is the most established tumor marker of hepatocellular carcinoma (HCC), but one of its limitations is non-specificity. Many studies have demonstrated that alpha-fetoprotein-L3 (AFP-L3) is more specific than AFP in the early diagnosis and prognosis of HCC. However, there is a lack of knowledge about the post-hepatectomy profiles of serum AFP and AFP-L3 values in HCC patients. To identify the profiles after surgical resection of HCC, we analyzed the correlation between the profiles and postoperative HCC recurrence or survival, and evaluated their utility in predicting postoperative therapeutic efficacy and prognosis. METHODS: From August 2003 to December 2004, 318 patients with positive serum AFP who had received surgical resections were enrolled in this study. Preoperative and postoperative serum AFP and AFP-L3 levels were measured simultaneously and regularly, and their postoperative profiles during a long term follow-up were recorded and summarized. RESULTS: A high ratio of AFP-L3 to total AFP was shown to correlate with pathologic features of aggressiveness. The overall 1-, 3-, and 5-year recurrence rates of the whole series were 28% 57%, and 84%, and the overall survival rates were 86%, 61% and 33%, respectively. The changes of serum AFP and AFP-L3 after hepatectomy for HCC were classified into 3 groups (group A: AFP-L3 undetectable; group B: AFP-L3 10%; and group C: AFP-L3 >10%). Patients with positive postoperative AFP-L3had significantly earlier recurrence than those with negative results. The overall survival was significantly shorter in the positive groups than in the groups negative for postoperative ***: Post-hepatectomy changes in serum AFP and AFP-L3 levels occurred in three distinct patterns, which were closely correlated with HCC recurrence and patient survival with different prognostic values.

关键词： hepatocellular carcinoma hepatectomy alpha-fetoprotein alpha-fetoprotein-L3

Increased frequency of the CTLA-4 +49A>G polymorphism in Chinese patients with colorectal cancer compared to healthy controls

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Increased frequency of the CTLA-4 +49A>G polymorphism in Chi...

全国临床免疫检验研讨会暨第六届全国临床免疫学术会议

作者： peng qi~1,Can-ping Ruan~2,Tong-hai Dou~3,hao wang~4,Fei-guo Zhou~5,Xin-yun xu~2,xing Gu~1,Yun-peng Zhao~1,chun-fang gao~(*1) 5.department of laboratory medicine,eastern hepatobiliary hospital,second military medical university,shanghai 200438,china 6.department of General Surgery,changzheng hospital,second military medical university,shanghai 200003,china 7.Institute of Genetics,Fudan university,shanghai 20043,china 8.department of laboratory medicine,changzheng hospital,second military medical university,shanghai 200003,china 9.department of Hepatic Surgery,eastern hepatobiliary hospital,second military medical university,shanghai 200438,china.

Objective Colorectal cancer is one of the most common malignancies in the world and a multipathway *** T-lymphocyte antigen-4(CTLA-4) is a potent immunoregulatory molecule that suppresses antitumor response by down-regulating T-cell *** most studied +49A>G polymorphism of CTLA-4 gene has been associated with various kinds of *** aim was to investigate the association between this genetic variant and the risk and progression of colorectal cancer in Chinese. Method We conducted a case-control study of 124 colorectal cancer cases and 407 healthy *** was extracted from blood specimens,and +49A>G polymorphism in the CTLA-4 gene was genotyped by polymerase chain reaction-ligation detection reaction(PCR-LDR).Results In our study group,the frequency of AG or GG or carrying at least one G allele at position +49 was significantly different in colorectal cancer patients and the control group,indicating that the risk of CRC was significantly higher among subjects with the AG or GG genotype or carrying at least one G allele at position +49 than among the subjects with the AA ***,we observed no association between CTLA-4 +49A>G polymorphism and the progression of CRC including tumor stage,tumor grade,status of lymph node and distant ***,the CTLA-4 +49A allele was in non-significantly higher numbers in CRC patients with distant *** Our results suggested that CTLA-4 +49A>G polymorphism was associated with an increased risk of colorectal cancer,but this polymorphism did not play an important role in Chinese patients with CRC.

关键词： Colorectal cancer cytotoxic T-lymphocyte antigen-4 single-nucleotide polymorphism case-control study individual susceptibility

Long non-coding RNA high expressed in hepatocellular carcinoma(IncRNA-HEIH) facilitates tumor growth through enhancer of zeste homolog 2

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Long non-coding RNA high expressed in hepatocellular carcino...

遗传学进步促进粮食安全与人口健康高峰论坛

作者： Fu yang1,ling Zhang1,Xi-song Huo1,Ji-hang Yuan1,Dan xu1,Sheng-xian Yuan2,Nan Zhu3,Wei-ping Zhou2,Guang-shun yang3,Yu-zhao wang1,Jing-li Shang1,chun-fang gao4,Feng-rui Zhang5,fang wang1,Shu-han Sun1 department of medical Genetics,second military medical university,shanghai,china 2 The Third department of Hepatic Surgery,eastern hepatobiliary hospital,second military medical university,shanghai,china 3 The second department of Hepatic Surgery,eastern hepatobiliary hospital,second military medical university, shanghai,china 4 department of laboratory medicine,eastern hepatobiliary hospital,second military medical university,shanghai,china 5 department of hepatobiliary Surgery,Hebei Cancer hospital,Hebei medical university,Hebei,china.

Hepatocellular carcinoma(HCC) is one of the most common human cancers worldwide, particularly in Southeast Asia and *** than 70-80%of HCC cases occur in high hepatitis B virus(HBV) endemic regions,and 50%of HCC cases worldwide are attributable to chronic infection with ***,the 5-year survival rate of HBV-related HCC patients remains poor,and approximately 600,000 HCC patients die each year despite recent advances in surgical techniques and medical *** previous studies identified many aberrantly expressed protein-coding genes in HCC,novel molecular markers that can help in early diagnosis and risk assessment are still urgently *** is of paramount importance to understand the relationships between clinical symptoms and molecular changes in HCC for developing new diagnosis and treatment strategies for HCC and improving the prognosis of diagnosed patients. The human transcriptome comprises not only large numbers of protein-coding messenger RNAs(mRNAs) but also a large set of non-protein coding transcripts that have structural,regulatory or unknown *** studies of small non-coding RNAs(18 to 200 nucleotides(nt)) have dominated the field of RNA biology in recent years,a surprisingly wide array of cellular functions has also been associated with long non-coding RNAs(IncRNAs).IncRNAs,tentatively defined as non-coding RNAs more than 200 nt in length,are characterized by the complexity and diversity of their sequences and mechanisms of action.A handful of studies have implicated IncRNAs in a variety of disease states,and altered IncRNA levels can result in aberrant expression of gene products that may contribute to cancer ***,there are only preliminary studies on the role of IncRNAs in HCC,and the overall pathophysiological contributions of IncRNAs to HCC remain largely unknown. In this study,we have identified non-overlapping signatures of a small number of IncRNAs that are aberrantly expressed in human HBV

关键词： hepatocellular carcinoma noncoding RNAs PRC2 IncRNAs cell cycle

Endoscopic management of biliary complications after orthotopic liver transplantation

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hepatobiliary & Pancreatic Diseases International 2006年第1期5卷 39-42页

作者： Yun-Sheng qin, Zhao-Shen Li, Zhen-xing Sun, Ren-Pei Wu, Na wang and Yin-Zhen Yao department of hepatobiliary Surgery, First Affiliated hospital, Zhejiang university, School of medicine, Hangzhou 310003, china department of Gastroenterology, Changhai hospital, second military medical university, shanghai 200433, china department of hepatobiliary Surgery First Affiliated Hospital Zhejiang University School of Medicine Hangzhou China.

BACKGROUND: Biliary complications are a serious problem in patients after liver transplantation and often require reoperation. This study was conducted to summarize the endoscopic diagnosis and management of biliary complications after orthotopic liver transplantation (OLT). METHODS: From December 2000 to November 2003, twelve endoscopic retrograde cholangiopancreatographies(ERCPs) were performed in 7 patients after OLT at Digestive Endoscopic Center of Changhai hospital in shanghai, china. The therapeutic maneuvers included endoscopic sphincterotomy (EST), biliary stent placement, balloon and basket extraction, irrigation, and nasobiliary tube placement. A retrospective study was made to determine the types of biliary tract complications after OLT. The success of ERCP and therapeutic maneuvers was also evaluated. RESULTS: Biliary tract complications including biliary stricture, biliary leak, biliary sludge, and stump leak of the cyst duct were treated respectively by endoscopic sphincterotomy with sludge extraction, stricture dilation or endoscopic retrograde biliary drainage. Two of the 3 patients with proximal common bile duct stricture were successfully treated with ERCP and stent placement. Four patients with anastomotic stricture and/without bile leak were treated successfully by dilation and stent placement or endoscopic nosobiliary drainage. No severe ERCP-related complications occurred. CONCLUSIONS: ERCP is an effective and accurate approach for the diagnosis of biliary tract complications after OLT, and placement of a stent is a safe initial treatment for biliary complications after liver transplantation.

关键词： liver transplantation biliary tract complications endoscopic retrograde cholangiopancreatography