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CD151 gene delivery increases eNOS activity and induces ECV304 migration, proliferation and tube formation

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Acta Pharmacologica Sinica 2007年第1期28卷 66-72页

作者： Zhen-zhong ZHENG Zheng-xiang LIU~2 Department of Cardiology,tongji Hospital,tongji medical college,Huazhong University of Science and Technology,Wuhan 430030,China Department of Cardiology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To investigate the effects of CD 151 on the activity of endothelial NO syn-thase(eNOS),and ECV304 migration,proliferation and tube ***:pAAV-CD 151 and pAAV-anti-CD 151 were constructed and used to transientlytransfect ECV304 mediated with Lipofectamine *** transfection,the ex-pression of CD 151 was measured by Western *** migration assay wasperformed using Boyden transwell;proliferation assay was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5,diphenyltetrazolium bromide(MTT)method,andtube formation test was examined on matrigel,eNOS activity was assayed by L-[~3H]citrulline production from L-[~3H]*** involvement of eNOS was ex-plored using an eNOS inhibitor(L-NAME)and the effects in the process ***:CD 151 promotes cell migration,proliferation and tube *** addition,CD151 increases eNOS ***,cell migration,prolifera-tion and tube formation induced by CD 151 are inhibited when L-NAME is used,which indicates that there is an involvement of eNOS in CD151-induced cellmigration,cell proliferation and tube ***:CD 151 promotesECV304 migration,proliferation and tube *** mechanism is that CD151increases eNOS *** result also suggests that eNOS is involved in theangiogenic effects of CD151.

关键词： CDI51 endothelial NO synthase(eNOS) cell migration cell proliferation tube formation

Alternation of AFP-mRNA level detected in blood circulation during liver resection for HCC and its significance

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World Journal of Gastroenterology 2002年第5期8卷 818-821页

作者： Xiao-Ping Chen Hong Zhao Hepatic Surgery Center,tongji Hospital,tongji medical college,Huazhong University of Science and Technology,Wuhan 430030,Hubei Province,China Xi-Ping Zhao Department of Clinic Immunology,tongji Hospital,tongji medical college,Huazhong University of Science and Technology,Wuhan 430030,Hubei Province,China Hepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China. chenxp_53@***

AIM: To investigate whether the current manipulation ofliver resection for hepatocellular carcinoma(HCC)causesdissemination of liver tumor cells into blood ***: Fifty patients with HCC, but without evidencesof metastases, were enrolled for the study. Samples ofperipheral blood before skin incision and after abdominalwall suture, and those of hepatic venous blood and portalvenous blood after liver parenchyma dissection,wereobtainedAFP-mRNA was extracted from the blood samplesand detected with reverse transcription polymerase chainreaction assay. The changes of its expression during theoperation were accessed by the ratio of fluorescence of AFPand β-actin primers-amplified ***: The rate of its expression before and afteroperation in peripheral blood, and during operation in portalvenous and hepatic venous blood was 42.0 %, 34.0 %, 42.0 %and 56.0 % respectively, There was no difference betweenthem. However, the ratio of fluorescence of AFP and β-actinprimers-amplified products from hepatic venous blood duringoperation is significantly higher than the other three typesof samples (P＜0.05).CONCLUSION: the current manipulation of liver resectionfor HCC can cause the releases of AFP mRNA-expressedcells from the liver, probably including tumor cells, intohepatic vein. These cells may spread into blood circulationand become the source of recurrence after operation.

关键词：肝癌肝切除术血液循环甲胎蛋白 mRNA 临床意义

Overcoming multi-drug resistance by anti-MDR1 ribozyme

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World Journal of Gastroenterology 2003年第7期9卷 1444-1449页

作者： Hai Wang Xiao-Ping Chen Fa-Zu Qiu The Hepatic Center of tongji Hospital,tongji medical college,Huazhong University of Science and Technology,Wuhan,430030,HuBei Province,China The Hepatic Center of tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China.

AIM: To reverse multidrug resistance (MDR) of HepG2 by anti-MDR1 hammerhead ***: We developed an anti-MDR1 hammerhead ribozyme and delivered it to P-gp-overproducing human hepatocarcinoma cell line HepG2 by a retroviral vector containing RNA polymerase Ⅲ promoter. We detected the expression of mdr1/Pgp and Rz in HepG2, HepG2 multidrugresistant cell line and HepG2 Rz-transfected cells by realtime RT-PCR, semi-quantitative RT-PCR and Western blot methods. Moreover, MTT assay was tested to detect sensitivity of these ribozyme-transfected cells, and Rhodamine123 (Rh123) applied to test the function of ***: The Rz- transfected HepG2 cells became doxorubicin-sensitive, concomitant with the decreases in MDR1 expression, P-gp amounts and efflux pump ***: The approaches using either retrovirus or liposome-mediated transfer of anti-MDR1 ribozyme may be selectively applicable to the treatment of MDR cells.

关键词：多药耐药性抗多药耐药核酶肝细胞癌逆转录病毒

Expression of Androgen Receptor in Meningiomas

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Journal of Huazhong University of Science and Technology(medical Sciences) 2001年第2期21卷 140-142页

作者：陈坚陈刚 Department of Neurosurgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan

In order to investigate the expression of androgen receptor in meningiomas and its relation to tumor proliferative potential, we examined the expression of AR and proliferating cell nuclear antigen by avidine biotin complex immunohistochemistry in 39 cases of meningiomas. Of the 39 cases of meningiomas, 20 showed positive AR immunoreactivity. The AR expression positivity rates were 31 % in benign meningiomas, 58 % in atypical meningiomas, 87.5 % in malignant meningiomas, respectively. In addition to the tumor cells, cells of microvascular endothelial proliferation were frequently AR positive. Malignant meningiomas had a significantly higher percentage of AR positive cells compared with atypical and benign meningiomas . The mean proliferating cell nuclear antigen labeling index was significantly higher in the malignant meningiomas when compared with atypical meningiomas and benign meningiomas . AR positive meningiomas had higher PCNA LI than AR negative meningiomas . The expression of AR in tumor tissues was significantly related with PCNA LI. These data indicated that AR in the meningiomas was correlated with histological grade and AR might participate in the growth of these tumors and tumor angiogenesis. The measurement of AR in these tumors may indirectly represent tumor growth potential.

关键词： meningiomas androgen receptor immunohistochemistry proliferating cell nuclear antigen angiogenesis

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Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro

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Asian Journal of Andrology 2003年第2期5卷 113-115页

作者： Ji-Hong LIU, Yang LI, Zheng-Guo CAO, Zhang-Qun YEDepartment of Urology, tongji Hospital, tongji medical college, Huazhong University of Science and Technology,Wuhan 430030, China Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Aim: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. Methods: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 癈. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. Results: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. Conclusion: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

关键词： dibutyryl cyclic AMP forskolin sperm motility

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Deguelin regulates nuclear pore complex proteins Nup98 and Nup88 in U937 cells in vitro1

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Acta Pharmacologica Sinica 2005年第10期26卷 1265-1273页

作者： Hong-li LIU~2, Yan CHEN~(2,3), Guo-hui CUI~2, Qiu-ling WU~2, Jing HE~2, Wei-hua CHEN~2, Jian-feng ZHOU~4 ~2 Department of Hematology, Union Hospital, tongji medical college, Huazhong University of Science and Technology, Wuhan 430022 ~4 Department of Hematology, tongji Hospital, tongji medical college, Huazhong University of Science and Technology, Wuhan 430030, China Department of Hematology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China Department of Hematology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim: To investigate the anticancer effects and the molecular mechanisms of deguelin on human U937 leukemia cells, and to explore the underlying mechanism regulating nucleoporin 98 （Nup98） and nucleoporin 88 （Nup88） in vitro. Methods: The effects of deguelin on the growth of U937 cells were studied by MTT assay. The effect of deguelin on the cell cycle of U937 cells was studied by using a propidium iodide method. The localization of the nuclear pore complex proteins Nup98 and Nup88 was investigated by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 and Nup88 in U937 cells was investigated by using flow cytometry and Western blot. Results: The prolif eration of U937 cells was inhibited in the deguelin-treated group, with a 24-h IC50value of 21.61 nmol/L and a 36-h IC50value of 17.07 nmol/L. U937 cells treated with deguelin had reduced percentages of cells in the G0/G1phase, whereas cells accumulated in the S and G2/M phases. Nup88 and Nup98 were found on both the nuclar and cytoplasmic sides of the U937 cells by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 was upregulated and that of the Nup88 protein was downregulated in U937 cells treated with deguelin. Conclusion: Deguelin is able to inhibit the proliferation of U937 cells by regulating the cell cycle such that cells are arrested at the S and G2/M phases, so that the proportion of cells in the G0/G1phase decreases. The antitumor effects of deguelin are related to upregulating the expression of Nup98 and downregulating the expression of Nup88 protein in U937 cells.

关键词： deguelin U937 cells nucleoporin Nup98 Nup88

The Relationship of CyclinD1 and Estrogen Receptor Expression in the Process of Proliferation and Metastasis in Breast Neoplasm

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Journal of Huazhong University of Science and Technology(medical Sciences) 2001年第3期21卷 231-232页

作者：王欣邹声泉 Department of Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan

The role of CyclinD1 and estrogen receptor (ER) in the process of proliferation and metastasis of breast neoplasm and their relationship were studied. The expression levels of CyclinD1 and ER in the tissue samples were detected by using flow cytometry and L SAB immunohistochemistry staining, respectively. The results showed that CyclinD1 and ER expression levels in breast cancer were significantly higher than in benign breast neoplasm (P0.05), but the CyclinD1 expression level in ER (+) group was significantly higher than in ER (-) group (P<0.05). It was concluded that CyclinD1 expression level might be obviously related with the proliferation and metastasis of breast neoplasm and ER.

关键词： breast cancer CyclinD1 flow cytometry estrogen receptor

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Immune tolerance induced by adoptive transfer of dendritic cells in an insulin-dependent diabetes mellitus murine model

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Acta Pharmacologica Sinica 2007年第1期28卷 98-104页

作者： Cheng-liang ZHANG Xiao-lei ZOU Jia-bei PENG Ming XIANG~2 Department of Pharmacology college of Pharmacy tongji medical college,Huazhong University of Science and Technology,Wuhan 430030,China Department of Pharmacology College of Pharmacy Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To investigate the effect and underlying mechanisms of immune-toleranceinduced by the adoptive transfer of bone marrow(BM)-derived dendritic cells(DC)in insulin-dependent diabetes mellitus(IDDM)***:The IDDMmodel was established by a low close of streptozotocin(STZ)in Balb/c *** subpopulations were generated from the BM cells with granulocyte-macroph-age colony-stimulating factor with or without *** purity and the Tcell stimulatory capability of DC were *** cells were used to modu-late autoimmune response in pre-diabetic *** glucose was examinedweekly;pancreas tissues were taken for histopathological analysis,and CD4~+ Tcells were isolated to detect lymphocyte proliferation by MTT assay and the ratioof CD4~+CD25+ T cells by fluorescence-activated cell sorting(FACS).The cytokinesecretion was determined by ELISA ***:Two DC subsets weregenerated from BM,which have phenotypes of mature DC(mDC)and immatureDC(iDC),*** level of blood glucose decreased significantly bytransferring iDC(P<0.01)rather than *** lymphocyte infiltration was ob-served in the islets,and pancreatic structure was *** vitro,proliferation oflymphocytes decreased and the proportion of CD4~+CD25~+ T cells increasedremarkably,compared with the mDC-treated groups(P<0.05),which were associ-ated with increased level of the Th2 cytokine and reduced level of the Th 1 cytokineafter iDC ***:Our data showed that iDC transfer was able toconfer protection to mice from STZ-induced *** immune-tolerance toIDDM may be associated with promoting the production of CD4~+CD25~+ T cellsand reducing regulatory Th2 responses in vivo.

关键词： dendritic cells insnlin-dependent diabetes mellitus streptozotocin immune tolerance regulatory T cells

Effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatoceUular carcinoma

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World Journal of Gastroenterology 2003年第11期9卷 2460-2463页

作者： Zhi-Xin Cao Xiao-Ping Chen Zai-De Wu, Hepatic Surgery Center, tongji Hospital, tongji medical college, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China Hepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China.

AIM: To study the effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatocellular ***: Twenty-six patients with HCC associated with cirrhosis were divided into hepatectomy with splenectomy group (splenectomy group, n=11) and hepatectomy without splenectomy group (non-splenectomy group, n=15). T lymphocyte subsets such as CD4, CD8, CD4/CD8, helper T (Th) lymphocyte cytokines such as interferon γ(IFN-γ),interleukin 2(IL-2), interleukin 10(IL-10) and white blood cell (WBC), platelet (PLT), total bilirubin (T-Bil) were measured and used as parameters to evaluate the effects of ***: There was no significant difference in CD4, CD8,CD4/CD8, IL2, IFN-γ, IL10, WBC, PLT, T-Bil levels between two groups before surgery. Two months after operation,the levels of CD4 (41.2 %±4.2 % vs 34.7 %±3.8 %), CD4/CD8 (1.7±0.2 vs 1.0±0.2), IFN-γ (102.3±15.9 pg/ml vs 86.5±14.8 pg/ml), IL-2(97.2±15.6 pg/ml vS77.6±14.5 pg/ml) were increased and those of CD8 (25.6±3.9 vs32.8 %±4.1%),IL-10 (56.9±10.4 pg/ml vs 72.6±15.3 pg/ml) were decreased in splenectomy groups as compared with those in nonsplenectomy group (P<0.05). WBC and PLT counts in the splenectomy group were 8.9±1.6×109 and 310±32×109,respectively, which were significantly higher than those in non-splenectomy group (3.7±1.4×109 and 104±41×109) respectively on the 14th post-operative day. T-Bil concentration in the splenectomy group (24±7 μmol/L) was significantly lower than that in the non-splenectomy group (37±13 μmol/L) on the 7th post-operative day (P<0.05).CONCLUSION: Splenectomy combined with hepatectomy for HCC associated with cirrhosis is helpful for the recovery of T-lymphocyte subsets and the maintenance of Th:L/Th2 cytokine balance.

关键词：肝细胞癌肝切除术肝硬化脾切除术临床研究