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Combination of CTLA4-FasL gene transfer and allogeneic bone marrow transplantation led to durable macrochimerism and donor-specific tolerance in mouse model

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Chinese Science Bulletin 2005年第20期50卷 2311-2317页

作者： FENG Yougang WANG Guangming HAO Jie LI Ailing YUAN Guohong LI Chong zeng fuqing XlE Shusheng Institute of Urology Union Hospital. Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China Department of Immunology Peking University Health Science Center Beijing 100083 China

Mixed hemopoietic chimerism is capable of inducing donor specific tolerance, thus eliminating the chronic immunosuppressive therapy following organ trans- plantation. As yet no safe and effective tolerance protocol is available for clinical implementation. Here we describe an alternative nonmyeloablative based strategy of using a single injection of recombination adenovirus vector encoding CTLA4-FasL fusing gene and donor bone marrow cells to promote durable mixed macrochimerism (>20% on 140 d). Chimeras exhibited robust donor-specific tolerance, as evi- denced by acceptance of fully allogeneic skin grafts (the mean survival time (MST)>200 d) and rejection of third- party skin grafts in a normal manner (MST<10 d). In this model, the frequencies of helper T lymphocyte precursor (HTLp) and cytotoxic T lymphocyte precursor (CTLp) were greatly reduced on day 14 after transplantation, suggesting that CTLA4-FasL led to rapid systemic peripheral tolerance to facilitate the bone marrow engraftment, while both HTLp and CTLp remained at low level only in recipient mice with mixed chimerism on day 140 after transplantation, demon- strating that long-term skin grafts tolerance was associated with stable mixed chimerism, and central deletion of donor specific T cell may be the main mechanism for tolerance maintenance.

关键词：老鼠动物模型骨髓移植嵌合现象免疫抑制力成活率

Protective Effect of Magnesium Isoglycyrrhizinate on Ethanol-Induced Testicular Injuries in Mice

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The Journal of Biomedical Research 2010年第2期24卷 153-160页

作者： Yuanqiao He fuqing zeng Qing Liu Wen Ju Houju Fu Hua Hao Lulu Lia Yifeng Xie Department of Urology Huazhong University of Science and Technology Wuhan 430022 Hubei Province China Department of Obstetrics and Gynecology Huazhong University of Science and Technology Wuhan 430022 Hubei Province China Department of Pathophysiology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China Department of Integrated Chinese Medicine and Western Medicine Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Hubei Province China

Objective： Ethanol treatment induces an increase in oxidative stress. As licorice compounds are potent antioxidants, our aim was to examine whether magnesium isoglycyrrhizinate attenuated lipid peroxidation, the major end-point of oxidative damage resulting from ethanol administration. Methods： Four groups（18 animals in each group） of male Kunming mice were used. The first group served as control and received 0.4 ml normal saline daily for 18 days orally. The second group of mice was given 56% ethanol at 16 ml/kg body weight per day for 18 days orally. The third group was given the same dose of ethanol and administrated magnesium isoglycyrrhizinate （15 mg/kg.d, i.p.） for 18 days. The fourth group was given the same dose of ethanol and administrated with magnesium isoglycyrrhizinate （45 mg/kg.d, i.p.） for 18 days. Twenty four hours after 9 days or 18 days of treatment the mice were sacrificed using 10% chloral hydrate. Sperm counts and motility in the epididymis were assessed. The lipid peroxidation and antioxidants of testicular mitochondria were also determined. The pathological changes of testicle tissue of the mice were observed by light microscopy. Results： Magnesium isoglycyrrhizinate effectively prevented the ethanol-induced seminiferous epithelium disorganization and degeneration of Sertoli cells and germ cells. Sperm counts and motility of the magnesium isoglycyrrhizinate treated groups were higher than those of the alcohol treated group, but were lower than those of the control group. The drug exhibited an ability to counteract ethanol induced oxidative challenge as it effectively reduced testicular malondialdehyde （MDA） and increased the activities of superoxide dismutase and glutathione peroxidase. Conclusion： Magnesium isoglycyrrhizinate is able to inhibit the ethanol-induced lipid peroxidation and has a protective effect against testicular oxidative injury.

关键词： ethanol oxidative damage testicular injury magnesium isoglycyrrhizinate

Construction of Smac gene-carrying and human uroplakin Ib promoter-Regulated Genetic Vector and its Expression

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Journal of Nanjing Medical University 2006年第5期20卷 275-278页

作者： Zhongxing Zhang fuqing zeng Guiyi Liao Xianghui Yue Department of Urology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

Objective： To construct an eukaryotic expression vector that contains Smac gene, which is regulated by human Uroplakin Ib （UpIb） promoter. Methods： For the directionality of Smac expression in the transitional cell carcinoma of bladder, internal CMV and T7 promoter sequences in eukaryotic expression vector pcDNA3.1-Smac were replaced with UpIb promoter to construct a new plasmid. The plasmid DNA was identified by gel electrophoresis after being double digested at respective sites, and then the sequence was analyzed. The expression of Smac mRNA and protein in BIU87 cell line were detected after the transfection by using the newly constructed vector. Results： The Smac gene-carrying and UpIb promoter-regulated eukaryotic expression vector pcDNA3-UpIb-promoter-Smac was successfully constructed. The expression of Smac mRNA was approximately increased by 2.1 times and the expression of Smac protein was increased in about 71% BIU87 cells. Conclusion： The new vector can be effectively expressed in bladder cancer cells and be of great significance for bladder cancer-targeted gene therapy.

关键词： Smac Uroplakin Ib promoter eukaryotic expression vector

Overview of Kidney Transplantation: A Single Centre Report from China

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Open Journal of Organ Transplant Surgery 2016年第2期6卷 7-11页

作者： Kiran Jang Kunwar Li Heng fuqing zeng Zhendi Wang Wuhan Union Hospital of China Tongji Medical College of Huazhong University of Science & Technology Wuhan China

Introduction: Worldwide, End Stage Renal Disease (ESRD) is one of the leading disease with prolong morbidity. Kidney transplantation offers the best solution for the problem. The shortage of donor kidney is even bigger problem due to transplantation being one of the routine procedures. The use of deceased donor definitely increases the pool of donor with excellent immediate and long-term follow-up proven results. Aim: The aim is to analyze and summarize the outcome of Kidney transplantation. Methods & Materials: A total of 78 cases of Kidney Transplantation were selected for the study and categorized as: Group I—41 (living Donor), Group II—23 (DCD) & Group III—15 (DBD). Perspective study was done with clean data recorded & maintained pre-operatively, post-operatively and follow-up from Jan 2011 to Dec 2015 in our hospital. Post-operative graft status, complications and at least 1-year follow-up were area of main focus. Results: All patients underwent successful kidney transplantation. In Group I, the number of living donor kidney transplantation is 41 whereas in Group II (DCD) & III (DBD), the number of deceased donor transplantation is 23 and 15 respectively. The Normal functioning of graft (NGF) was 38 (87.8%), 16 (69.6%) & 11 (73.3%) in Group I, II & III respectively along with Poor Graft function (PGF) in Group I—4 (9.7%), II—5 (21.7%) & III—2 (13.3%) managed by continuing dialysis. Delayed graft function (DGF) was noted I-1 (2.4%), II-2 (8.6%) & III-1 (6.6%) in respective group, which returned to normal function post intervention. Therefore, 1st year graft survival was >93% [(Group I (97.6%), Group II (95.6%) & Group III (93.3%) respectively]. Manageable surgical complication were found in Group I—8 (19.5%), Group II—5 (21.7%) & Group III—2 (13.3%) like hematoma, hydronephrosis, leakage except one emboli related nephrectomy of transplanted kidney & one pneumonia led death in Group II. The overall survival was greater than 90% [(Group I (97.6%), Group

关键词： Kidney Transplantation (KTx) ESRD—End Stage Renal Disease Deceased Donor (DCD—Donor after Cardiac Death DBD—Donor after Brain Death)

Construction of Smac gene-containing and human prostate specific antigen promoter-regulated vector and its expression

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Journal of Nanjing Medical University 2007年第3期21卷 134-138页

作者： Yu Wu fuqing zeng Liang Wang Yanbo Wang Guiyi Liao Department of Urology Union Hospital Tongji Medical College Huazhong University of Science and Technotogy Wuhan 430022 China

To construct an eukaryotic expression vector containing Smac gene and study the expression efficiency and specificity of prostate specific antigen（PSA） enhancer/promoter in a possible targeted gene therapy scheme for prostate cancer. Methods： PSA enhancer （PSAE） and promoter （PSAP） sequences were amplified using PCR method. CMV and T7 promoters were deleted from pcDNA3.1-Smac and replaced by the two specific fragments to generate pPSAE-PSAP-Smac. After transfection into different cell lines, the status of cells was observed. And then, we determined the relative concentration of Smac mRNA in RT-PCR. Results： The recombinant plasmid of pPSAE-PSAP-Smac was successfully constructed. And only the prostate cancer cell line PC-3 was suppressed after transfection with pPSAE-PSAP-Smac. However, other nonprostate lines were not. Moreover, the concentration of Smac mRNA regulated by PSA promoter and enhancer was higher in comparison to the CMV promoter-driven control vectors. Conclusion： An expression vector containing the Smac gene （based on elements of the PSA gene regulatory sequences） has been developed and shown to function in prostate cancer cell lines which provides a solid platform for launching clinical studies.

关键词： prostate specific antigen enhancer promoter Smac gene therapy

Detection of Smac expression in bladder cancer and its clinical significance

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Journal of Nanjing Medical University 2006年第1期20卷 48-51页

作者： Guiyi Liao fuqing zeng Xianghui Yue Liang Wang Fangmin Cheng Department of Urology. Union Hospital Tongji Medical College Huazhong University of Sciences and Technology Wuhan 430022 China

Objective： To detect the expression of second mitochondria-derived activator of caspase （Smac） in bladder cancer and discuss its clinical significance. Methods： Smac was detected in 15 specimens of normal bladder epithelium and 72 specimens of bladder cancer by reverse transcription-polymerase chain reaction （RT-PCR） and immunohistochemistry at the level of gene and protein, respectively. Results： The differences of both Smac protein and mRNA expressions between normal mucous membrane of bladder and grade i bladder cancer had no statistical significance （ P 〉 0.05 ）. The expressions of Smac protein and its mRNA in bladder cancer decreased gradually with the advance of bladder cancer （ P 〈 0.01 and P 〈 0.05, respectively）. In invasive bladder cancer, the expressions of Smac protein and its mRNA were higher than those in superficial bladder cancer （ P 〈 0.01）. Conclusions： Normal bladder epithelium has high expression of Smac while bladder cancer has low expression of Smac. The expression of Smac is closely related to the grade and stage of bladder cancer. Detection of Smac expression helps to judge the grade and stage of bladder cancer and Smac gene might become a valid target for gene therapy of bladder cancer.

关键词： Smac bladder neoplasm cancer apoptosis

The inhibitory effects of siRNA expression vector on CXCR4 expression in prostate carcinoma cell lines

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Journal of Nanjing Medical University 2006年第2期20卷 104-108页

作者： Yuefeng Du Yifei Xing fuqing zeng Peng Lu Xianyin Liu Department of Urology Union hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022.China

Objective： To investigate the inhibitory effects of RNAi （ RNA interference, RNAi） expression vector on CXCR4 expression in prostate carcinoma cell lines. Methods： Small interference RNA （siRNA） expression vectors for CXCR4 gene were constructed and transfected into prostate carcinoma cell lines（PC-3m and LNCaP）with liposomes. T expression of CXCR4 was detected by RT-PCR and western blot. Results： T expression of CXCR4 mRNA and protein in the PC-3m and LNCaP cells was reduced by RNAi expression vectors. The inhibitory rate of CXCR4 mRNA expression in the PC-3m cells was 87.81% ± 10.20% ,56.10% ± 9.32% at the 24th hour and the 48th hour, compared with 56.93% ±8.78% ,49.24% ± 11.23% in LNCaP cells. The inhibitory rate of the expression of CXCR4 protein was 64.71% ± 6.68% ,58.66% ± 11.56% respectively. Conclusion： The expression of CXCR4 gene can effectively be inhibited by RNAi expression vectors.

关键词： RNA interference(RNAi) CXC receptor 4 expression vector RT-PCR western blot

Emergency Treatment of Testicular Torsion and Postoperative Follow-up:71 Case Reports

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Emergency Treatment of Testicular Torsion and Postoperative ...

双侧上尿路结石并发肾功能不全术后急性肾衰的处理

中华医学会男科学分会第十三次全国男科学术会议

作者： Feng Pan Chaohui Zhu Bing Li Zili Pang Yajun Xiao fuqing zeng Department of Urology Union HospitalTongji Medical CollegeHuazhong University of Science and Technology

This study summarize the experience in the treatment oftesticular torsion;which promote early diagnosis and improve therapeutic effects.A retrospective analysis was conducted in 71 patients with testicular torsion that was treated in our hospital from May 2007 to April *** age of the patients ranged from 16 day to 34 years *** the patients showed unilateral testicular torsion;which presented on the left side of 43 cases and on the right side of 28 *** course of the disease varied between 3 hours to 30 ***-operative

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临床泌尿外科杂志 2009年第5期24卷 349-350,355页

作者：陈浩鞠文曾甫清华中科技大学同济医学院附属协和医院泌尿外科武汉430022

目的:探讨治疗双侧上尿路结石并发肾功能不全术后急性肾衰的方法。方法:对54例双侧上尿路结石并发肾功能不全术后急性肾功能衰竭(ARF)患者的治疗方法进行回顾性分析,根据不同病情分别行急诊输尿管镜碎石(URL)和输尿管逆行插管或经皮肾造... 详细信息

目的:探讨治疗双侧上尿路结石并发肾功能不全术后急性肾衰的方法。方法:对54例双侧上尿路结石并发肾功能不全术后急性肾功能衰竭(ARF)患者的治疗方法进行回顾性分析,根据不同病情分别行急诊输尿管镜碎石(URL)和输尿管逆行插管或经皮肾造瘘(PCN)后二期碎石。所有病例监测血肌酐(Cr)和尿素氮(BUN)。结果:术后全部患者血Cr和BUN进行性下降,恢复到正常43例,氮质血症6例,尿毒症4例,死亡1例。结论:双侧上尿路结石并发肾功能不全应早期诊断,选择恰当的外科治疗时机和方法,一旦术后出现ARF,要立即明确原因,进行治疗。

关键词：上尿路结石肾功能不全术后急性肾功能衰竭