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High efficient mammalian expression and secretion of a functional humanized single-chain Fv/human interleukin-2 molecules

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World Journal of Gastroenterology 2006年第24期12卷 3859-3865页

作者： Yue-Chun Shen xue-haowang xiao-ming wang Zao-lai chen Xi-Ping Shen Chao-chen zhao jun li division of Cardiology Department of InternalMedicine The University of Michigan Medical School 1150 WestMedical Center Drive Ann Arbor M148109 United States division of hepatic surgery Department of General Surgery The First Affiliated Hospital of Nanjing Medical University 300 Guangzhou Rd Nanjing 210029 Jiangsu Province China

AIM： To construct and produce a recombinant bispecific humanized single-chain Fv （sFv） /Interleukin-2 （IL-2） fusion protein by using mammalian cells. METHODS： The sFv/IL-2 protein was genetically engineered, and transfected to mammalian cells to determine whether the mammalian protein folding machinery can produce and secrete active sFv/IL-2 with high efficiency. RESULTS： The fusion protein was constructed and high efficiently expressed with yields up to 102 ±4.2 mg/L in culture supernatant of the stably transfected 293 cell line. This recombinant fusion protein consisted of humanized variable heavy （VH） and light （VL） domains of monoclonal antibody （mAb） 520C9 directed against the human HER-2/neu （c-erbB2） proto-oncogene product p185, and human IL-2 connected by polypeptide linker. The fusion protein was shown to retain the immunostimulatory activities of IL-2 as measured by IL- 2-dependent cell proliferation and cytotoxicity assays. In addition to its IL-2 activities, this fusion protein also possessed antigen-binding specificity against p185, as determined by indirect EliSA using p185 positive SKOV 3ip1 cells. CONCLUSION： The large-scale preparation of the recombinant humanized sFv antibody/IL-2 fusion protein is performed with 293 cells. The recombinant humanized sFv antibody/IL-2 fusion protein may provide an effective *** targeting therapeutic doses of IL-2 to p185 positive tumors without increasing systemic toxicity or immunogenicity.

关键词： Interleukin-2 Humanization Antibody Fusion protein HER-2/neu

Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

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World Journal of Gastroenterology 2023年第18期29卷 2798-2817页

作者： liu Shi li Zhou ming Han Yu Zhang Yang Zhang xiao-xue Yuan Hong-Ping Lu Yun wang xue-liang Yang chen liu jun wang Pu liang Shun-Ai liu xiao-Jing liu jun cheng Shu-Mei lin department of Infectious Disease Medicine The First Hospital Affiliated to Xi’an Jiaotong UniversityXi’an 710061Shaanxi ProvinceChina china-Japan Friendship hospital Department of Infectious Disease China-Japan Friendship HospitalBeijing 100029China Institute of Infectious Diseases Beijing Ditan HospitalCapital Medical UniversityBeijing 100015China The division of liver Diseases Beijing Ditan HospitalCapital Medical UniversityBeijing 100015China Institute of liver Diseases Beijing Pan-Asia Tongze Institute of Biomedicine Co.LtdBeijing 100015China department of Rehabilitation Medicine The First Affiliated Hospital of Xi’an Jiaotong UniversityXi’an 710061Shaanxi ProvinceChina Beijing Key Laboratory of Emerging Infectious Diseases Peking University Ditan Teaching HospitalBeijing 100015China

BACKGROUND hepatic fibrosis is a serious condition,and the development of hepatic fibrosis can lead to a series of ***,the pathogenesis of hepatic fibrosis remains unclear,and effective therapy options are still *** group identified hepatitis C virus nonstructural protein 3-transactivated protein 1(NS3TP1) by suppressive subtractive hybridization and bioinformatics analysis,but its role in diseases including hepatic fibrosis remains ***,additional studies on the function of NS3TP1 in hepatic fibrosis are urgently needed to provide new targets for *** To elucidate the mechanism of NS3TP1 in hepatic fibrosis and the regulatory effects of calcitriol on *** Twenty-four male C57BL/6 mice were randomized and separated into three groups,comprising the normal,fibrosis,and calcitriol treatment groups,and liver fibrosis was modeled by carbon tetrachloride(CCl4).To evaluate the level of hepatic fibrosis in every group,serological and pathological examinations of the liver were ***-β1 was administered to boost the in vitro cultivation of LX-2 ***3TP1,α-smooth muscle actin(α-SMA),collagen I,and collagen Ⅲ in every group were examined using a Western blot and real-time quantitative polymerase chain *** activity of the transforming growth factor beta 1(TGFβ1)/Smad3 and NF-κB signaling pathways in each group of cells transfected with pcDNA-NS3TP1 or siRNA-NS3TP1 was *** statistical analysis of the data was performed using the Student’s t *** NS3TP1 promoted the activation,proliferation,and differentiation of hepatic stellate cells(HSCs)and enhanced hepatic fibrosis via the TGFβ1/Smad3 and NF-κB signaling pathways,as evidenced by the presence of α-SMA,collagen I,collagen Ⅲ,p-smad3,and p-p65 in LX-2 cells,which were upregulated after NS3TP1 overexpression and downregulated after NS3TP1 *** proliferation of HSCs was lowered after NS3TP1 interference and elevated after

关键词： Nonstructural protein 3-transactivated protein 1 Calcitriol liver fibrosis hepatic stellate cells Mouse model TGFβ1/Smad3 NF-κB Signaling pathway

Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma(2022 version)

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Journal of Clinical and Translational Hepatology 2023年第7期11卷 1553-1564页

作者： Han wang jun chen Xin Zhang Xia Sheng xiao-Yan Chang Jie chen Min-Shan chen Hui Dong Guang-Jie Duan He-Ping Hu Zhi-Yong Huang Wei-Dong Jia xiao-Qing Jiang Dong Kuang Shan-Shan li Zeng-Shan li Chang-li Lu Shu-Kui Qin xue-Shan Qiu li-Juan Qu Chun-Kui Shao Feng Shen Guo-ming Shi Su-Sheng Shi Yu-jun Shi Hui-Chuan Sun xiao-Dong Teng Bin wang Zhan-Bo wang Tian-Fu Wen Jia-Mei Yang Qiao-Qiao Yang Sheng-Long Ye Hong-Fang Yin Zhen-Gang Yuan Jing-Ping Yun Feng-lin Zang Hong-Qi Zhang li-Hong Zhang Jing-Min zhao Jian Zhou Wei-Xun Zhou Jia Fan xiao-Ping chen Wan Yee Lau Yuan Ji Wen-ming Cong Chinese Society of liver Cancer of Chinese Anti-Cancer Association Digestive Disease Group of Chinese Society of Pathology,Chinese medical Association Chinese Society of Pathology of Chinese Anti-Cancer Association hepatic surgery Group of Chinese Society of surgery,Chinese medical Association Biliary Tract Tumor Committee of china Anti-Cancer Association Chinese Society of Clinical Oncology Idepartment of Pathology Eastern Hepatobilary Surgery HospitalNaval Medical UniversityShanghaiChina department of Pathology the Affiliated Drum Tower HospitalNanjing University Medical SchoolNanjingJiangsuChina department of Pathology Zhongshan HospitalFudan UniversityShanghaiChina department of Pathology Minhang HospitalFudan Uni-versityShanghaiChina department of Pathology Peking Union Medical College HospitalChinese Academy of Medical Sci-ences and Peking Union Medical CollegeBeijingChina department of liver surgery Sun Yat-Sen University Cancer CenterGuangzhouGuangdongChina department of Pathology The First Affiliated HospitalArmy Medical UniversityChongqingChina department of Hepatobiliary Medicine Eastern Hepatobiliary Surgery HospitalNaval Medical UniversityShanghaiChina hepatic surgery Center Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWu-hanHubeiChina department of General surgery The First Affliated Hospital of USTCDivision of Life Science and Medi-cineUniversity of Science and Technology of ChinaHefeiAnhuiChina department of Biliary surgery I Eastern Hepatobil-iary Surgery HospitalNaval Medical UniversityShanghaiChina department of Pathology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubeiChina department of Pathology The First Afiliated Hospital of Zhengzhou UniversityZhengzhou UniversityZhengzhouHenanChina department of Pathology Xijing Hospi-talAir Force Medical UniversityXi'anShaanxiChina department of Pathology West China HospitalSichuan UniversityChengduSichuanChina Cancer Center of Jinling hospital Nanjing University of Chinese MedicineNanjingJiangsuChina department of Pathology The First Affiliated HospitalChina Medical UniversityShenyangLiaoningChina department of Pathology The 900 Hospital of the Chinese People's Liberation Army Joint Logistics TeamFuzhouFujianChina department of Pathology The Third Affiliated HospitalSun Yat-Sen UniversityGuangz

Intrahepatic cholangiocarcinoma(iCCA)can originate from the large bile duct group(segment bile ducts and area bile ducts),small bile duct group(septal bile ducts and interlobular bile ducts),and terminal bile duct group(bile ductules and canals of Hering)of the intrahepatic biliary tree,which can be histopathological corresponding to large duct type iCCA,small duct type iCCA and iCCA with ductal plate malformation pattern,and cholangiolocarcinoma,*** challenge in pathological diagnosis of above subtypes of iCCA falls in the distinction of cellular morphologies,tissue structures,growth patterns,invasive behaviors,immunophenotypes,molecular mutations,and surgical *** these reasons,this expert consensus provides nine recommendations as a reference for standardizing and refining the diagnosis of pathological subtypes of iCCA,mainly based on the 5th edition of the World Health Organization Classification of Tumours of the Digestive System.

关键词： Intrahepatic cholangiocarcinoma Pathology Histological type Immunohistochemistry Molecular variation

A practical community-based response strategy to interrupt Ebola transmission in sierra Leone, 2014–2015

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Infectious Diseases of Poverty 2016年第1期5卷 670-679页

作者： Zhong-Jie li Wen-xiao Tu xiao-Chun wang Guo-Qing Shi Zun-Dong Yin Hai-jun Su Tao Shen Da-Peng Zhang Jian-Dong li Shan Lv Chun-li Cao Rui-Qian Xie Hong-Zhou Lu Rong-Meng Jiang Zheng Cao Zhi-Jie An Lei-Lei li Jie Xu Yan-Wen Xiong Wei Zang Wei Zhang Hong-Wei Zhang Wen-Sen chen Hua ling Wen Xu Jian Cai Huan-Jin Luo xue-Sheng Xing Can-jun Zheng Qiang Wei Xin-Xu li Mei li Hai Jiang li-Quan Deng ming-Quan chen Xiang Huo Feng Xu xue-Hui lai Xi-chen Bai Long-Jie Ye Jian-Yi Yao Wen-Wu Yin Jiao-Jin Sun lin xiao Fu-Qiang liu xiao-Qiang liu Hong-Wei Fan Zeng-Qiang Kou Ji-Kun Zhou hao Zhang Da-Xin Ni Thomas TSamba Qun li Hong-Jie Yu Yu wang xiao-Feng liang division of Infectious Disease Key Laboratory of Surveillance and Early-warning on Infectious DiseaseChinese Center for Disease Control and PreventionBeijingChina Public Health Emergency Center Chinese Center for Disease Control and PreventionBeijingChina National Center for AIDS/STD Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina Chinese Field Epidemiology Training Program Chinese Center for Disease Control and PreventionBeijingChina National Immunization Program Chinese Center for Disease Control and PreventionBeijingChina Bureau of Disease Prevention and Control National Health and Family Planning Commission of the People’s Republic of ChinaBeijingChina National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina National Institute of Parasitic Disease Chinese Center for Disease Control and PreventionShanghaiChina Chinese Center for Health Education BeijingChina department of Infectious Diseases Shanghai Public Health Clinical CenterShanghaiChina Beijing Ditan hospital Capital Medical UniversityBeijingChina Health News BeijingChina National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and PreventionBeijingChina department of Training Chinese Center for Health EducationBeijingChina Center for Infectious Diseases Beijing Youan HospitalCapital Medical UniversityBeijingChina Infection Management The First Affiliated Hospital of Nanjing Medical UniversityNanjingChina Chongqing Municipal Center for Disease Control and Prevention ChongqingChina Yunnan Provincial Center for Disease Control and Prevention KunmingChina division of Infectious Disease Control and Prevention Zhejiang Provincial Center for Disease Control and PreventionHangzhouChina Guangdong Provincial Center for Disease Control and Prevention GuangzhouChina division of Acute Infectious Disease Control and Prevention Hubei Provincial Center for Disease Control and PreventionW

Background:The Ebola virus disease spread rapidly in West Africa in 2014,leading to the loss of thousands of *** engagement was one of the key strategies to interrupt Ebola transmission,and practical community level measures needed to be explored in the field and tailored to the specific context of ***:first,community-level education on Ebola virus disease(EVD)prevention was launched for the community’s social mobilizers in six districts in Sierra Leone beginning in November ***,from January to May of 2015,in three pilot communities,local trained community members were organized to engage in implementation of EVD prevention and transmission interruption measures,by involving them in alert case report,contact tracing,and social *** epidemiological indicators of transmission interruption in three study communities were ***:A total of 6016 community social mobilizers from 185 wards were trained by holding 279 workshops in the six districts,and EVD message reached an estimated 631680 *** three pilot communities,72 EVD alert cases were reported,with 70.8%of them detected by trained local community members,and 14 EVD cases were finally *** tracing detected 64.3%of EVD *** median duration of community infectivity for the cases was 1 *** secondary attack rate was 4.2%,and no third generation of infection was *** health worker was infected,and no unsafe burial and noncompliance to EVD control measures were *** community-based measures were modeled to reduce 77 EVD cases,and the EVD-free goal was achieved four months earlier in study communities than whole country of Sierra ***: The community-based strategy of social mobilization and community engagement was effective in casedetection and reducing the extent of Ebola transmission in a country with weak health system. The successfullypractical experience to reduce the risk of Ebola transmission in the

关键词： Ebola virus disease Community engagement Health education Outbreak control

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Cellular & Molecular Immunology 2021年第4期18卷 1005-1015页

作者： xuezhen Zeng Jingying Zhou Zhewen Xiong Hanyong Sun Weiqin Yang Myth T.S.Mok Jing wang Jingqing li Man liu Wenshu Tang Yu Feng Hector Kwong-Sang W ang Shun-Wa Tsang King-Lau Chow Philip Chun Yeung John Wong Paul Bo-San lai Anthony Wing-Hung Chan Ka Fai To Stephen Lam Chan Qiang Xia Jing xue xiao chen jun Yu Sui Peng Joseph Jao-Yiu Sung ming Kuang Alfred Sze-Lok cheng School of Biomedical Sciences The Chinese University of Hong KongHong Kong SARChina lnstitute of Precision Medicine The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina department of liver surgery The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina department of Pharmacy The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina department of Anatomical and Cellular Pathology The Chinese University of Hong KongHong Kong SARChina department of liver surgery Ren Ji HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina department of Gastroenterology The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina division of life Science Hong Kong University of Science and TechnologyHong Kong SARChina department of surgery The Chinese University of Hong KongHong Kong SARChina State Key Laboratory of Translational Oncology The Chinese University of Hong KongHong Kong SARChina department of Clinical Oncology The Chinese University of Hong KongHong Kong SARChina State Key Laboratory of Oncogenes and Related Genes Stem Cell Research CenterRen Ji HospitalSchool of MedicineShanghai Cancer InstituteShanghai Jiao Tong UniversityShanghaiChina department of Medicine and Therapeutics The Chinese University of Hong KongHong Kong SARChina State Key Laboratory of Digestive Disease The Chinese University of Hong KongHong Kong SARChina Clinical Trial Unit The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina Cancer Center The First Affiliated HospitalSun Yat-sen UniversityGuangzhouChina

The liver is an immunologically tolerant organ and a common metastatic site of multiple cancer *** a role for cancer cell invasion programs has been well characterized,whether and how liver-intrinsic factors drive metastatic spread is incompletely ***,we show that aberrantly activated hepatocyte-intrinsic cell cycle-related kinase(CCRK)signaling in chronic liver diseases is critical for cancer metastasis by reprogramming an immunosuppressive *** an inducible liverspecific transgenic model,we found that CCRK overexpression dramatically increased both B16F10 melanoma and MC38 colorectal cancer(CRC)metastasis to the liver,which was highly infiltrated by polymorphonuclear-myeloid-derived suppressor cells(PMNMDSCs)and lacking natural killer T(NKT)*** of PMN-MDSCs in CCRK transgenic mice restored NKT cell levels and their interferon gamma production and reduced liver metastasis to 2.7% and 0.7%(metastatic tumor weights)in the melanoma and CRC models,***,CCRK activated nuclear factor-kappa B(NF-κB)signaling to increase the PMN-MDSC trafficking chemokine C-X-C motif ligand 1(CXCL1),which was positively correlated with liver-infiltrating PMN-MDSC levels in CCRK transgenic ***,CRC liver metastasis patients exhibited hyperaaivation of hepatic CCRK/NF-κB/CXCL1 signaling,which was associated with accumulation of PMN-MDSCs and paucity of NKT cells compared to healthy liver transplantation *** summary,this study demonstrates that immunosuppressive reprogramming by hepatic CCRK signaling undermines antimetastatic *** findings offer new mechanistic insights and therapeutic targets for liver metastasis intervention.

关键词： Cell cycle related kinase liver metastasis liver immune microenvironment myeloid-derived suppressor cell natural killer T cell