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Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第3期19卷 198-202页

作者： LIU Shangqing , LIU Xiufang , ZOU Ping , SONG Shanjun , WANG Bianming Institute of Hematology, Xiehe hospital, tongji Medical University, Wuhan 430022 Institute of Hematology Xiehe Hospital Tongji Medical University Wuhan 430022

An immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding bone marrow sections obtained from 10 patients with myelodysplastic syndrome (MDS) was presented. The results of qualitative and quantitative immunohistochemical analysis are as follows: (1) Labile antigens of T lymphocytes were well preserved, thus allowing analysis of distribution of T lymphocyte subsets in situ ; (2) the average number of T 3, T 4 and T 8 lymphocyte of the diffuse infiltrate was about 2 %, 0.4 %, 0.5 %, respectively, of all nucleated cells in bone marrow, and T 4/T 8 of T cells were below 1.0 in patients with MDS; (3) there were cases of RAS showing T lymphocyte aggregation in bone marrow, but no patient exhibited progressive refractory anemia with excess of blasts(RAEB) and RAEB in transformation (RAEBT). These findings indicated that the immunological abnormalities are of importance in the evaluation of pathogenesis and prognosis of MDS.

关键词： immunohistochemistry plastic embedding of bone marrow distribution of T cell subsets in situ myelodysplastic syndrome

Cloning,Eukaryotic Expression of Human ISG20 and Preliminary Study on the Effect of Its Anti-HBV

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2008年第1期28卷 11-13页

作者：郝友华杨东亮 Division of Clinical Immunology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

Human ISG20 gene was cloned and the effect of its anti-HBV was primarily studied. The ISG20 gene was amplified from HeLa cells by RT-PCR and recombinant vector expressing ISG20 was constructed by genetic engineering. The overexpression of ISG20 in HepG2 cells was detected by Western blot and the levels of secretion of HBs antigen and HBe antigen tested by ELISA. The results showed that：（1） Sequence of ISG20 cloned was consistent to that published in Genebank; （2） Recombinant vector expressing ISG20 could be expressed in HepG2 cells by transfection; （3） The overexpression of ISG20 protein could reduce the levels of the secretion of HBs antigen and HBe antigen in transfected HepG2 cells. It was suggested that the overexpression of recombinant ISG20 in culture cells could reduce the synthesis of HBV proteins.

关键词： human ISG20 effect of anti-HBV

Laparoscopic Assisted Surgery for Crohn's Disease an Initial Experience and Results

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2000年第4期20卷 332-335页

作者：栾晓军 Department of Internal Medicine Tongji Hospital Tongji Medical University Wuhan 430030

The inflammatory process associated with Crohn's disease often makes dissection difficult, even in open surgery. The aim of this study was to assess the technical feasibility, safety and indica- tion of laparoscopic assisted procedures performed in patients with Crohn's disease. Records of pa- tients undergoing surgical operation for Crohn's disease from 1993 to 1998 at our hospital were re- viewed. Intestinal resection was performed laparoscopically in 24 patients (LAP) and by open tech- nique in 23 patients(OPEN). No significant differences existed as to age, gender, body-mass-index and previous surgery. In the laparoscopic group, seven operations (CON) were converted to open la- parotomy (29 %) because of large inflammatory mass and/or fistula. The mean intraoperative blood loss was significantly higher in CON- and OPEN-groups than in LAP-group (P<0. 01). Major com- plications occurred only in one patient who underwent laparoscopic assisted high anterior resection. Patients who underwent laparoscopic operation tolerated p. o. liquids sooner than patients who under- went open surgery (median: 2 vs. 5 day, P<0. 05). Compared with the CON- and OPEN-groups, patients in LAP-group had lower analgesic requirements (median: 3 vs. 6 and S day, P<0. 01). The median postoperative length of stay was significantly shorter in LAP-group than in OPEN-group (median: 11 vs. 14 day, P<0. 05). Our study showed that LAP is technically feasible for Crohn's disease. The preoperative correct diagnosis and selection of indications are very important, because the laparoscopic mobilization and resection may be difficult or impossible in patients with large fixed masses, multiple complx fistulas, or recurrent Crohn'sdisease.

关键词： Crohn's disease laparoscopic assisted surgery indication

Specific cellular immunity and antitumor responses in C57BL/6 mice induced by DNA vaccine encoding murine AFP

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Hepatobiliary & Pancreatic Diseases International 2004年第3期3卷 440-443页

作者： Geng Tian, Ji-Lin Yi and Ping Xiong Department of Oncology, Shenzhen Second hospital,Shenzhen 518035, China tongji hospital and Depart-ment of Immunology , tongji Medical College, Huazhong Uni-versity of Science and Technology, Wuhan 430030, China Department of Oncology Shenzhen Second Hospital Shenzhen 518035 China. geng_tian707@***

BACKGROUND: The inoculation of plasmid DNA encod-ing tumor-associated antigens is a novel and powerful stra-tegy for antitumor vaccination. This study was designed toconstruct the DNA vaccine of mouse AFP and to observethe specific cellular immunologic responses and the antitu-mor responses in mice induced by this ***: The murine AFP gene was amplified by RT-PCR from total RNA extracted from Hepal-6 cells andcloned into the vector pcDNA3.1 to construct *** DNA vaccine was identified by restriction enzymeanalysis, sequencing and expression. EL-4 ( mAFP) wasdeveloped by stable transfection of EL-4 cells with *** frequency of cells producing IFN-γ in splenocytes har-vested from the mice immunized with the DNA vaccine byintramuscular injection was measured by enzyme linkedimmunospot (ELISPOT). The mice immunized with theDNA vaccine were inoculated with EL-4 (mAFP) cells inback to observe the inhibitory effect of the immunizationon tumor. On the other hand, blood samples were collect-ed from the immunized mice to check the functions of theliver and ***: The murine AFP gene was successfully clonedby RT-PCR. Results from restriction enzyme analysis, se-quencing and expression showed that the DNA vaccine wassuccessfully constructed. The expression of mAFP mRNAin EL-4 (mAFP) was confirmed by RT-PCR. The resultsof ELISPOT showed that the number of IFN-γ-producingcells of the pmAFP vaccine group was significantly higherthan that of other groups (P <0.01). The tumor volume inthe pmAFP vaccine group (1042. 42 ± 123. 71 mm3 ) wassignificantly smaller than that in other groups (P<0.01).The function of mouse liver and kidney in each group ***: The successfully constructed DNA vaccineof AFP can induce specific cellular immunologic responsesand significant antitumor reponses in mouse and has no im-pact on the function of mouse liver and kindey.

关键词： DNA vaccine α-fetoprotein mouse immunotherapy

Non-radioactive Hybridization in Microwells Using Enzym eLinked Im m uneSorbentAssay forDetection ofRT-PCR-Am -plified CK19-and CEA-m RNA

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Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第3期19卷 181-184,189页

作者： ZHONG Xiaoyan 1, Sepp Kaul 2, Astrid Eichler 2, Gunther Bastert 2 1 Department of Obstetrics and Gynecology, tongji hospital, tongji Medical University, Wuhan 430030 2 Laboratory of Oncology, Department of Obstetrics and Gynaecology, Univ Department of Obstetrics and Gynecology Tongji Hospital Tongji Medical University Wuhan 430030

PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT PCR amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non radioactive hybridization was performed in a streptavidin coated microwell with digoxigenin labeled PCR products and with biotin labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19 , CEA RT PCR products. These results were proven by sequencing.

关键词： CK19 CEA RT PCR ELISA non radioactive hybridization

Immunohistochemical Study of HLA-DR Antigen in Endometrial Tis-sue of Patients with Endometriosis

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2002年第1期22卷 60-61页

作者：刘义罗丽兰赵海波 Department of Obstetrics and Gynecology Xiehe HosPital Huazhong University of Science and Technology Wuhan 430022 Department of Obstetrics and Gynecology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

In order to evaluate the expression of HLA DR antigen in glandular cells in eutopic and ectopic endometrium in patients with endometriosis, 19 infertile patients with endometriosis were analyzed immunohistochemically by labelled streptavidin biotin (LSAB) method. Nineteen infertile patients without endometriosis were studied as controls. The results showed that the expression of HLA DR antigen in the glandular cells in both eutopic and ectopic endometrium was increased significantly as compared with that in the controls ( P <0.01). It is likely that aberrant expression of HLA DR antigen in endometriotic tissue is involved in abormal immunogenesis of endometriosis.

关键词： HLA DR antigen immunohistochemistry endometriosis

Inhibiting Apoptosis of CTLL-2 Cells to Enhance Their GVL Effects via Anti-Fas Ribozyme

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 559-565页

作者： Min ZHANG, Fang LIU, Lin-Bo LIU, Yong YOU, Zhi-Chao CHEN, and Ping ZOUInstitute of Hematology, Union hospital, tongji Medical College, Huazhong University of Science & Technology, Wuhan 430022, China Institute of Hematology Union Hospital Tongji Medical College Huazhong University of Science & TechnologyWuhan 430022 China

To investigate the inhibition role of anti-Fas hammerhead ribozyme on fas expression and Fas-mediated apoptosis of CTL cell line CTLL-2 cells, the cDNA of an anti-Fas hammerhead ribozyme was synthesized, its expression plasmid was constructed and transfected into CTLL-2 cells by electroporation. fas expression of CTLL-2 cells was detected by RT-PCR and Western blot. CTLL-2 cell viability was measured using MTT assay when co-cultured with mouse T cell leukemia cell line EL4 cells that highly expressed Fas ligand (FasL). Meanwhile, caspase-3 proteolytic activity was detected, and cell apoptosis was measured by flow cytometry and Hochest-PI double staining. Killing activity of CTLL-2 cells was detected by lactate dehydrogenase (LDH) releasing assay in vitro. Results showed that the expression of both Fas mRNA and protein in CTLL-2 cells were decreased after transfection of anti-Fas ribozyme. Compared with mock- transfected group and mutant ribozyme-transfected group, viability of CTLL-2 cells co-cultured with EL4 cells was increased significantly and cells killing activity was enhanced after transfected with anti-Fas ribozyme, while the caspase-3 activity and apoptosis rate was significantly decreased. The results demonstrated anti-Fas ribozyme could efficiently cleave Fas and inhibit Fas-mediated apoptosis of CTLL-2 cells to improve their viability. Our study made a basis for enhancing CTLL-2 cells anti-leukemia effect in DLI.

关键词： Hammerhead ribozyme anti-Fas CTLL-2 cells apoptosis tumor immunity

Low and Maternal-specific Expression of p57^(KIP2) in Hydatidiform Mole and Its Clinical Implication

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2002年第2期22卷 121-122,157页

作者：熊雅丽曹阳李红发 Department of Obstetrics and Gynecology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Department of Ophthalmology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 Department of Obstetrics and Gynecology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

Summary: In situ hybridization was applied to locate and detect the expression of p57 KIP2 in hydatidiform mole (5 cases of partial hydatidiform mole and 18 cases of complete hydatidiform mole) and normal villi (23 cases). The positive signals of p57 KIP2 expression were analyzed by HPIAS-1000 Image-Analysis System. p57 KIP2 was highly expressed in normal villi but showed distinct low expression in hydatidiform mole (P<0.01). Furthermore, the locus of low expression of p57 KIP2 accorded with the place where lesion of trophoblast occurred. Detection of p57 KIP2 made it possible to study the genetics of hydatidiform mole at the transcriptional level. Low expression of p57 KIP2 could be a molecular marker in hydatidiform mole and a target for therapy.

关键词： p57 KIP2 hydatidiform mole in situ hybridization

Experimental Study of Mouse Cytomegalovirus Infected M ice

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2002年第3期22卷 260-264页

作者：崔雯董永绥方峰 Department of Pediatrics Tongji Hospital Tongji Medical University Huazhong University of Science and Technology Wuhan

In order to investigate the human cytomegalovirus (HCMV) infection, the mouse cytomegalovirus (MCMV) infected mice were experimentally studied. 6 to 8 week old female BALB/C mice with immunosuppression were selected to undergo the MCMV inoculations: intracranial inoculation and peritoneal inoculation. MCMV of the infected mice in various organs and tissues were detected by using  gal staining and in situ nucleic acid hybridization assay. The pathological cha nges were observed in HE staining paraffin embedded sections. It was found that all the MCMV infected mice showed the retardation of growth and development, an d feather looseness. Both intracranial inoculation of 10 4 PFU viruses or perit oneal inoculation of 10 6 PFU viruses resulted in the pathological changes, to some extent, of various organs and tissues in the mice. The pathological changes in liver were consistent with the amount of  gal staining positive cells, in dicating the liver lesions were mainly caused by viral proliferation. It was als o found that the viruses in the immunosuppressed mice subjected to intracranial inoculation could spread to whole body organs, while the viruses in the immunosu ppres sed mice subjected to intrapeitoneal inoculation couldn't spread to the brain, s uggesting blood brain barrier could prevent the virus from spreading to the bra in.

关键词： mouse cytomegalovirus infection

Investigation of the Distribution and Changes of VLDLR Subtype in Fibrotic Cardiac Muscles

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2000年第4期20卷 297-300页

作者：杨光田鲍力赵金平屈伸 Department of Emergency Tongji Hospital Tongji Medical University Wuhan Department of Biochemistry Tongji Medical University Wuhan Department of Cardiothoracic Surgery Tongji Hospital Tongji Medical University Wuhan

Very low-density lipoprotein receptor (VLDLR) is the major receptor with which cells can uptake the triacylglycerol from blood. It is divided into two subtypes according to presence of O- linked sugar domain located in the VLDLR receptor immediately outside of the membrane. Type Ⅰ VLDLR contains the O-link domain, while type Ⅱhas no such domain. The type ⅠVLDLR are mainly found on the surface of human myocardial cells. The result of our quantitative polymerase chain reaction on the normal and fibrotic cardiac muscles showed that both subtypes and expression level of VLDLR on the myocardial cell surface did not vary significantly between the normal and the fibrotic cardiac muscles despite the presence of malfunction due to fibrosis. This finding suggests that fibrosis doesn't exert significant influence on the subtype and the expression of VLDLR on the sur- face of myocardial cells. Such inconsistence with the changes found in other fibrotic tissues is awaiting further studies.

关键词： myocardial cells cardiac muscle fibrosis very low-density lipoprotein receptor (VLDLR)