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| 1 | Effect of a cancer vaccine prepared by fusions of hepatocarcinoma cells with dendritic cells显示文摘AIM To prepare a cancer vaccine (H22-DC) expressing high levels of costimulatory molecules based on fusions of hepatocarcinoma cells ( H22 ) with dendritic cells (DC) of mice and to analyze the biological characteristics and induction of specific CTL activity of H22-DC.``METHODS DCs were isolated from murine spleen by metrizamide density gradient centrifugation, purified based on its characteristics of semi-adhesion to culture plates and FcR , and were cultured in the medium containing GM. CSF and IL-4. A large number of DC were harvested. DCs were then fused with H22 cells by PEG and the fusion cells were marked with CDllc MicroBeads. The H22-DC was sorted with Mimi MACS sorter. The techniques of cell culture, immunocytochemistry and light microscopy were also used to test the characteristice of growth and morphology of H22-DC in vitro. As the immunogen, H22-DC was inoculated subcutaneously into the right armpit of BALB/C mice, and their tumorigenicity in vive was observed. MTT was used to test the CTL activity of murine spleen in vitro.``RESULTS DC cells isolated and generated were CD1 lc +cells with irregular shape, and highly expressed CD80,CD86 and CD54 molecules. H22 cells were CDllc cells with sphericel shape and bigger volume, and did not express CD80, CD86 and CD54 molecules. H22-DC was CDllc+ cells with bigger volume, being spherical, flat or irregular in shape, and highly expressed CD80, CD86 and CD54 molecules, too. H22-DC was able to divide and proliferate in vitro, but its activity of proliferation was significantly decreased as compared with H22 cells and its growth curve was flatter than H22 cells. After subcutaneous inoculation over 60 days, 1-12_2-DC showed no tumorigenecity in mice, which was significantly different from control groups (P< 0.01 ) . The spleen CTL activity against H22 cells in mice implanted with fresh H22-DC was significantly higher than control groups (P< 0.0l).``CONCLUSION H22-DC could significantly stimulate the specific CTL activity of murine spleen, which suggests that the fusion cells have already obtained the function of antigen presenting of parental DC and could present H22specific antigen which has not been identified yet, and H22-DC could induce antitumor immune response; although simply mixed H22 cells with DC could stimulate the specific CTL activity which could inhibit the growth of tumor in some degree, it could not prevent the generation of tumor. It shows that the DC vaccine is likely to become a helpful approach in immunotherapy of hepstocarcinoma. | Juan Zhang~1 Jin-Kun Zhang~2 Shao-Hong Zhuo~3 Hai-Bin Chen~2 1 Clinical Laboratory,The First Affiliated Hospital of Shantou University Medical College,Shantou 515041,Guangdong Province,China2 Cancer Pathology Laboratory,Shantou University Medical College,Shantou 515031,Guangdong Province,China3 Department of Gastroenterology,Third Municipal Hospital of Shantou,Shantou 515073,Guangdong Province,China | 2001 | World Journal of Gastroenterology2001，7，5： | 26 |
| 2 | Antitumor activities of human dendritic cells derived from peripheral and cord blood显示文摘AIM: To observe the biological specialization of humanperipheral blood dendritic cells (DC) and cord blood derivedDC and its effects on effector cells killing humanhepatocarcinoma cell line BEL-7402 in vitro.METHODS: The DC biological characteristics were detectedwith immunohistochemical and MTT assay. Two antitumorexperimental groups are: peripheral blood DC and cordblood DC groups. Peripheral blood DC groups used LAKcells as the effector cells and BEL-7402 as target cells, whilecord blood DC groups used CTL induced by tumor antigentwice pulsed DC as effector cells and BEL-7402 as targetcells, additional peripheral blood DC and cord blood DC areadded to observe its stimulating activities to effector cells.The effector's cytotoxicity to tumor cells were detected withneutral red colorimetric assay at two effector/target ratios of5:1 and 10: 1.RESULTS: Peripheral blood DC and cord blood DC highlyexpressed HLA-ABC, HLA-DR, HLA-DQ, CD54 and S-100protein. The stimulating activities to lymphocyteproliferation were compared between experimental groups(DC added) and control group (no DC added). In sixexperiment subgroups, the DC/lymphocyte ratio wassequentially 0.25: 100, 0.5: 100, 1: 100, 2: 100, 4: 100 and 8:100, A values(x± s) were 0.75396± 0.009, 0.84916± 0.010,0.90894± 0.012, 0.98371 ± 0.007, 1.01299 ± 0.006 and 1.20384± 0.006 in peripheral blood DC groups and 0.77650 ± 0.005,0.83008± 0.007, 0.92725 ± 0.007, 1.05990 ± 0.010, 1.15583 ±0.011, 1.22983 ± 0.011 in cord blood DC groups. A value was0.59517 ± 0.005 in control group. The stimulating activitieswere higher in experimental groups than in control group ( P< 0.01 ), which were increased when the DC concentrationwas enlarged ( P < 0.01 ). Two differently derived DCs hadthe same phenotypes and similar stimulating activities ( P >0.05). In peripheral blood DC groups, the cytotoxicity (x ±s) of the LD groups (experimental groups) and L groups(control group) was 58.16% ± 2.03% (5: 1), 46.18% ±2.25% (10:l) and 38.13% ± 1.29% (5: 1) and 65.40% ±1.56% (10: 1), respectively; in cord blood DC groups, TDgroups (experimental groups ) and T groups (controlgroups) were 69.71% ± 2.33% (5: 1 ), 77.64% ± 1.94% ( 10:1) a. nd 56.89% ± 1.82% (5:1) and 60.99% ± 1.42% (10:1),respectively. The cytotoxicity activities were enhanced withincreased effector/target ratio ( P < 0.01 ). At the sameeffector/target ratio, the cytotoxicity of experimental groupswas higher than that of control groups ( P < 0.01 ). Thecytotoxicity of cord blood DC groups were higher than ofperipheral blood DC groups ( P < 0.01 ).CONCLUSION: Peripheral blood DC and cord blood DC aremature DCs in morphology and function, both can enhancethe effector cell killing activities to hepetocarcinoma cells.DC pulsed with tumor antigen can induce ligher specific CTLactivity than unpulsed DC. | Jin-Kun Zhang Jun Li Hai-Bin Chen Jin-Lun Sun Yao-Juan Qu Juan-Juan Lu Cancer Pathology Laboratory,Shantou University,Medical College,Shantou 515031,Guangdong Province,China | 2002 | World Journal of Gastroenterology2002，8，1： | 18 |
| 3 | Early diagnosis for colorectal cancer in China显示文摘AIM: To review the present studies on early diagnosis ofcoiorectal cancer.METHODS: The detective rate for early cancer is 1.7%-26. 1 % based on various statistical data, with much higherdetective rate in endoscopy. Since early cancer meansinvasion involved in the mucosa or submucosa, thediagnosis can only be made when the invasive depth isidentified. Pathological tissue materials from bothsurgical operation or endoscopic resection are suitable forearly cancer evaluation.RESULTS: Incidence of polyp malignancy is 1.4%~20.4%. The various constitutive proportion of polyps mayexplain the different rates. Malignant incidence is higherin adenomatous polyps, that for villous polyps can reach21 .3%-58.3%. Type Ⅱ early stage of colorectal carcinomais rarely reported in China. it is shownd that majority ofthem were not malignant, most of type lla being adenomaor hyperplasia, and llb being inflammatory and llc mightbe the isolated ulcers. The occurrence of malignancy oftype Ⅱ is far lower than that of polypoid lesion. In China,the qualitative diagnosis and classification of neoplasmgenerally adopted the WHO standard, including surgicalexcision or biopsies. There is impersonal evaluationbetween colorectal pre-malignancy and cancer. Theformer emphasizes the dysplasia of nuclei and gland,while the latter is marked with cancer invasion. Diagnosisof early stage colorectal cancer in endoscopy is made withtoo much caution which made the detective rate muchlower. Mass screening for asymptomatic subjects andfollow-up for high risk population are mainly used to findthe early stage colorectal cancer in China. Fecal occultblood test is also widely made as primary screening test,galactose oxygenase test of rectal mucus (T antigen),fecal occult albumin test are also used. The detective rateof colorectal cancer is 24-36.5 per 105 mass population.CONCLUSION: Although carcinoma associated antigen inblood or stool, microsatellite DNA instability for high riskfamilial history, molecular biology technology for stooloncogene or antioncogene, telomerase activity andexfoliative cytological examination for tumor marker, areutilized, none of them is used in mass screening by now. | Ya-LiZhang Zhen-SuZhang 等 | 2002 | World Journal of Gastroenterology2002，8，1： | 14 |
| 4 | Construction and selection of the natural immune Fab antibody phage display library from patients with colorectal cancer显示文摘AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer.METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer.RT-PCR was used to amplify the heavy chain Fd and light chain к and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected.RESULTS: The amplified fragments of Fd and к gained by RT-PCR were about 650bp. Fd and к PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 106. The libraries were enriched about 120-fold by 3 cycles of adsorption-elution- multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer.CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer. | Bao-Ping Wu~1 Bing Xiao~1 Tian-Mo Wan~1 Ya-Li Zhang~1 Zhen-Shu Zhang~1 Dian-Yuan Zhou~1 Zhuo-Sheng Lai~1 Chun-Fang Gao~2 1 Institute for Digestive Diseases,Nanfang Hospital,Guangzhou 510515,Guangdong Province,China2 Surgical Department of Colon and Rectum,150 Central Hospital,Luoyang 471031,Henan Province,China | 2001 | World Journal of Gastroenterology2001，7，6： | 9 |
| 5 | Effect of DPC4 Gene on Invasion and Metastasis of Colorectal Carcinoma Cells显示文摘在侵略和转移 ofcolorectal 癌房间上调查 DPC4 基因的效果, DPC4 的表示在 colorectaltumors 的六十三件样品和颜色的七个案例被检测表面的 mucosa.The 表示 DPC4 的肿瘤的生物行为被评估(包括肿瘤阶段,区别度和转移) .pcDNA3.1-DPC4 原生质标志被构造并且转了进不稳定地表示 DPC4.The 房间模型( DPC4^+-HCT116 )的 HCT116 房间表示 DPC4 被获得。与 HCT116 和 pcDNA3.1-HCT116 房间相比, DPC4^+-HCT116 房间的双倍工资显然被变长(P<0.01 ) , DPC4^+-HCT116cells 的 apoptosis 率显著地被增加(P<0.01 ) ,克隆的效率,房间 adherency,迁居和 DPC4^+-HCT116 房间的侵略能力显然被扔(P<0.01 ) 。癌症小瘤的数字在腹的洞显著地被减少,裸体老鼠的肝接种 withDPC4^+-HCT116 房间。 MMP-9 和 MMP-2 的活动被明胶 zymography.In 比较与 HCT116 和 pcDNA3.1-HCT116 房间检测, MMP-9 的活动在 DPC4^+-HCT116cells.Therefore 被减少, DPC4 表示的下面规定可以与 ofcolorectal carcinoma.DPC4 可以禁止的 carcinogenesis 被联系由制止生长并且导致 apoptosis 的结肠癌房间的增长,并且表面的癌 cells.MMP-9 可以是的颜色的侵略和转移下游的目标基因之一由 DPC4 调整了。 | De-Sheng XIAO Ji-Fang WEN Jing-He LI Kuan-Song WANG Zhong-Liang HU Jian-Hua ZHOU Zheng-Hao DENG Ying LIU | 2006 | Acta Biochimica et Biophysica Sinica2006，38，12： | 8 |
| 6 | Influence of granulocyte-macrophage colonystimulating factor and tumor necrosis factor on anti-hepatoma activities of human dendritic cells显示文摘INTRODUCTIONDendritic cells (DCs) play a key regulatory role inantitumor immunity,especially in its immuneaccessory role via MHC-Ⅰ molecules.We haverecently reported that DCs were able to enhance thekilling activity of Lymphokine and PHA activatedkiller (LPAK) cells in vitro.In the presentstudy,we evaluated the effects of GM-CSF andTNF upon antitumor activities of freshly | Jin Kun Zhang Jin Lun Sun Hai Bin Chen Yang Zeng Yao Jun Qu | 2000 | World Journal of Gastroenterology2000，6，5： | 8 |
| 7 | In situ expression and significance of B7 costimulatory molecules within tissues of human gastric carcinoma显示文摘AIM: To explore the role and significance of costimulatory molecules B7H1,B7H2 and ICOS within tissues of human gastric carcinoma and the possible mechanisms in tumor escape.
METHODS: mRNA expressions of costimulatory molecules including B7H1,B7H2,ICOS and B7-1 in tissues of human gastric carcinoma were investigated by in situ hybridization using digoxigenin-labeled oligonucleotide-probes. The tissue of chronic gastric ulcer was used as a control. All data were analyzed by SPSS statistic software.
RESULTS: At the site of gastric carcinoma, mRNA expression levels of B7H1, B7H2 and ICOS were much higher than that of B7-1. Their mRNA positive expression indexes were 0.512±0.333, 0.812±0.454, 0.702±0.359 and 0.293±0.253, respectively. The positively stained cells were mainly tumor infiltrating lymphocytes (TILs), and some tumor cells. The difference between them was greatly significant P<0.005. The mRNA expression levels of four molecules were not correlated to the pathological grade and matastasis of gastric carcinoma.
CONCLUSION: ICOS-B7H costimulatory pathway may be predominant at the site of gastric carcinoma. B7-1mRNA might be the basis of ICOS-B7H interaction. ICOS-B7H interaction induces the production of IL-10 which inhibits the antitumor immune responses. Therefore, it is supposed that ICOS-B7H costimulatory pathway may be involved in the negative regulation of cell-mediated immune responses. | Bao-ShanSu Li-LiWang Xiao-LiChen Xu-DongCao An-JingKang Kang-MinWang | 2003 | World Journal of Gastroenterology2003，9，6： | 7 |
| 8 | Arterial chemotherapy of 5-fluorouracil and mitomycin C in the treatment of liver metastases of colorectal cancer显示文摘AIM: Regional chemotherapy using hepatic arterycatheters is a good method of treating patients withcolorectsl cancer liver metastases. We investigated thesurvival of patients with liver metastases fromcolorectal cancer using 5-fluorouracil (5-FU) andmitomycin C Cthrough implantable hepatic arterialinfusion pert. METHODS: Seventy-five patients with inoperable livermetastases from colorectal cancer were includedbetween March, 1992 and November, 2001. We placedimplantable hepatic arterial catheter (HAC) port bylaparotomy. 5-FU, 1 000 mg/m2/d continuous infusionfor five days every four weeks, was delivered in thehepatic arterial catheter through the port. MitomycinC, 30 mg/m2/d infusion in the first day every cyclethrough the port. Response to the treatment wasevaluated by serial determinations of plasma CEA andimaging techniques consisting of computerizedtomography and sonography of liver.RESULTS: Sixty-eight were performed hepatic arterychemotherapy and fifty-six were followed up amongseventy-five HAC patients. Twenty-six patients(46.4 %)have responded and 4 complete remission wereachieved. Eight patients (14.3 %) had stable livermetastases. Twenty-two patients (39.3 %) wereprogressed with increased tumor size and number.Twenty-nine patients(51.8% ) had a decreased serumCEA level, while 10 patients (17.9 %) were stable and17 patients (30.4 %) had an increased serum CEA level.There were no operative death in this series.Complications, which occurred in 18 patients (32.1%),were as followed: hepatic artery thrombosis in 11,Upper gastric and intestinal bleeding in 3, liver abscessin 1, pocket infection in 1, cholangitis in 1, and hepaticartery pseudo-aneurysm in one patient.CONCLUSION: Combined infusion of 5-FU and mitomycinC by hepatic artery catheter port is an effectivetreatment for liver metastases from colorectal cancer.The high response and lower complication rates provethe adjuvant treatment of colorectal cancer with thistreatment. | Lian-Xin Liu Wei-Hui Zhang Hong-Chi Jiang An-Long Zhu Lin-Feng Wu Da-Xun Piao Department of Surgery,the First Clinical College,Harbin Medical University,Harbin 150001,Heilongjiang Province,China Shu-Yi Qi Department of VIP,the First Clinical College,Harbin Medical University,Harbin 150001,Heilongjiang Province,China | 2002 | World Journal of Gastroenterology2002，8，4： | 7 |
| 9 | 大肠腺癌组织Survivin蛋白的表达意义显示文摘目的:检测凋亡抑制蛋白Survivin在大肠腺瘤、腺癌组织中的表达,观察其与细胞增生、凋亡的关系,探讨Survivin基因表达在大肠癌发生、发展过程中的作用.方法:应用DNA缺口未端标记技术和免疫组织化学S-P法,原位观察20例正常大肠黏膜组织、35例大肠腺瘤和60例大肠腺癌组织中的凋亡细胞和Survivin、Ki-67蛋白阳性表达.结果:Survivin蛋白在正常大肠黏膜组织中不表达,在大肠腺癌和腺瘤中阳性表达率分别为60.0%和17.1%,二者比较差异有显著性(P<0.01).Survivin蛋白表达与大肠腺癌分化程度呈负相关,与Dukes’分期和淋巴结转移无明显相关.大肠腺癌Ki-67标记指数(labelingindex,LI)为39.1±10.4%,较腺瘤22.3±6.2%显著增高(P<0.01),大肠腺癌Survivin阳性组Ki-67LI25.1±7.6%与Survivin阴性组19.7±5.8%比较差异有显著性(P<0.01).大肠腺瘤和腺癌细胞凋亡指数(AI)显著高于正常大肠黏膜(P<0.01),高分化癌凋亡指数显著高于低分化癌(P<0.05),大肠腺癌Survivin阳性组凋亡指数0.85±0.52%与Survivin阴性组1.25±0.58%比较差异有显著性(P<0.01).结论:Survivin基因在大肠腺癌组织中表达上凋,通过促进肿瘤细胞增生和抑制凋亡参与大肠癌的发生、发展过程.检测大肠组织Survivin蛋白表达,对预测癌变及判断预后有重要意义.Survivin基因为肿瘤的基因治疗提供一理想靶点. | 肖军 邓长生 朱尤庆 | 2003 | 世界华人消化杂志2003，11，5： | 6 |
| 10 | 结肠癌组织Smad4蛋白表达的意义显示文摘目的:探讨Smad4在结肠癌发生、发展过程中的作用方法:采用免疫组化S-P法检测70例正常结肠黏膜组织和结肠肿瘤组织中Smad4的表达. 结果:结肠癌(n=52)中Smad4的表达水平明显低于正常结肠黏膜(n=7),并与肿瘤分期、分化程度及转移有关(P<0.05). 结论:Smad4的低表达可能与结肠癌的发生、发展过程有关,并可能在结肠癌浸润和转移中发挥作用. | 肖德胜 李景和 傅春燕 文继舫 | 2004 | 世界华人消化杂志2004，12，5： | 6 |
| 11 | Regulatory effects of electro-acupuncture at Zusanfi on ir-SP content in rat pituitary gland and peripheral blood and their immunity显示文摘INTRODUCTIONIt has been reported in many studies that electro-acupuncture(EA)can positively regulate erythrocyticimmunity and T-lymphocytic subgroups.Nevertheless,its mechanism remains to be explored.In thepresent study,a multi-group,multi-stepped and multi-indexed observation was conducted on the effects of EA | Wei Gao Yu Xin Huang Hong Chen Da Yong Song Qin Li Wang Department of Gastroenterology Medical Affair Office,Tangdu Hospital,4th Military Medical University,Xi’an 710038,Shanxi Province,China Department of Gastroenterology,Chinese PLA General Hospital of Guangzhou Command Area,Guangzhou 510210,Guangdong Province,China | 2000 | World Journal of Gastroenterology2000，6，4： | 5 |
| 12 | Tumor necrosis factor-related apoptosis-inducing ligand gene on human colorectal cancer cell line HT29显示文摘AIM: To evaluate the therapeutic efficiency of Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) gene on human colorectal cancer cell line HT29.METHODS: Human embryonal kidney cells transformed by introducing sheared fragments of Ad5 DNA (293 cell)were used for amplification of adenoviral vectors: Ad/GTTRAIL,Ad/GT-Bax, Ad/GT-LacZ and Ad/PGK-GV16. Human colorectal cancer cell line HT29 was transfected with binary adenovirus-mediated TRAIL gene. Bax gene was used as positive control, LacZ gene was used as the vector control,and cells treated with PBS only were used as a mock control. The morphological changes, cell growth and apoptosis were measured by reversmicroscope, MTT method and flow cytometry.RESULTS: All adenoviral vectors titer determined by optical absorbency at A260nm were 1×1010 viral particle/ml(vp/ml).Obviously morphological changes of HT29 cells were observed when infected with Ad/GT-TRAIL, and these changes were much more obviously when Ad/PGK-GV16was coinfected. The cell suppression percentage and the percentage of apoptotic cells were 52.5 % and 16.5 %respectively when infected with Ad/GT-TRAIL alone, while combining with Ad/PGK-GV16, the growth of HT29 was suppressed by 85.2 % and the percentage of apoptotic cells was 35.9 %. It showed a significantly enhanced therapeutic efficiency with binary system (P<0.05).CONCLUSION: A binary adenoviral vector system provides an effective approach to amplify viral vectors that express potentially toxic gene, TRAIL. Ad/GT-TRAIL showed a significantly enhanced therapeutic efficiency for HT29 when coinfected with Ad/PGK-GV16. Ad/GT-TRAIL could induce apoptosis of HT29 and inhibit its growth. | Xiang-MingXu ChaoHe Xiao-TongHu Bing-LiangFang | 2003 | World Journal of Gastroenterology2003，9，5： | 3 |
| 13 | DPC4基因转染对结肠癌细胞生物学行为的影响显示文摘目的:研究DPC4基因转染对结肠癌细胞生物学行为的影响,探讨DPC4在结肠癌的发生和发展中的作用.方法:利用基因重组技术构建PcDNA3.1-DPC4质粒;利用脂质体介导转染技术和G418筛选得到稳定表达Smad4蛋白的DPC4+-SW620(PcDNA3.1-DPC4转染的SW620高转移性结肠癌细胞株)细胞模型;采用免疫组化S-P法和Westernblot检测细胞中Smad4的表达;生长曲线和平板克隆形成实验检测其生物学行为的改变;采用流式细胞术检测其S期细胞百分数(S%)和凋亡率.结果:成功构建了PcDNA3.1-DPC4质粒;DPC4+-SW620细胞的Smad4蛋白表达于细胞质及细胞核,以细胞质为主,且Smad4蛋白表达水平明显高于SW620细胞及PcDNA3.1-SW620细胞(PcDNA3.1转染的SW620细胞);DPC4+-SW620细胞的倍增时间为116h,较SW620细胞(31h,P<0.01)及PcDNA3.1-SW620细胞(29h,P<0.01)明显延长;DPC4+-SW620细胞的克隆形成率为(12%),明显低于SW620细胞(69%,P<0.01)及PcDNA3.1-SW620细胞(67%,P<0.01)的克隆形成率;与SW620细胞及PcDNA3.1-SW620细胞相比较,DPC4+-SW620细胞G0-G1期细胞百分数增多,S期细胞百分数明显减少(P<0.05);与SW620细胞及PcDNA3.1-SW620细胞相比较,DPC4+-SW620细胞的凋亡率较高.结论:成功地构建了质粒PcDNA3.1-DPC4,并转染SW620细胞株,得到稳定且表达Smad4蛋白明显增强的细胞模型;DPC4对结肠癌细胞增生的调控可能是通过DPC4抑制细胞生长和诱导细胞凋亡两方面作用实现的. | 肖德胜 文继舫 李景和 胡忠良 郑晖 傅春燕 | 2002 | 世界华人消化杂志2002，10，9： | 2 |
| 14 | 肝癌细胞H_(22)与树突状细胞杂交瘤苗的实验研究显示文摘目的:将肝癌H22细胞与树突状细胞(DC)相融合,研制杂交瘤苗,并观察瘤苗的生长特性、致瘤性及免疫活性.方法:将DC与肝癌H22细胞融合制备H22-DC融合细胞,磁式分选器分选融合细胞;观察融合细胞的生长特性和体内致瘤性;从Balb/C小鼠皮下接种融合细胞,实验设肿瘤等三组对照,每组各5只小鼠.MTT比色法检测小鼠脾CTL活性.结果:融合细胞在体外能分裂增生,但明显低于肿瘤细胞,无体内致瘤陛;活融合细胞免疫的小鼠,其脾CTL杀伤H22细胞的活性明显高于对照组小鼠(P<0.01).结论:融合细胞能诱导出特异性的CTL活性,可望成为肝癌免疫治疗的新途径. | 张娟 张锦堃 卓少宏 | 2004 | 世界华人消化杂志2004，12，2： | 2 |
| 15 | B7-1基因转染骨肉瘤细胞诱导抗骨肉瘤主动免疫的实验研究显示文摘目的:探讨B7-1基因转染骨肉瘤细胞能否诱导抗骨肉瘤主动免疫作用。方法:利用脂质体将B7-1真核表达载体pcDNA3-B7-1和空载体pcDNA3分别导入骨肉瘤细胞株LM8中,G418筛选出阳性克隆(分别命名为LM8/B7-1和LM8/pcDNA3),通过RT-PCR、Western blot和流式细胞仪检测B7-1基因与蛋白的表达;通过软琼脂克隆形成实验观察转基因细胞株LM8/B7-1的体外增殖能力;用LM8、LM8/B7-1和LM8/pcDNA3分别经腹腔免疫小鼠,得到腹腔浸润淋巴细胞和致敏脾细胞,MTT法检测其体外杀伤活力;将LM8、LM8/B7-1和LM8/pcDNA3细胞分别接种到C3H雄性小鼠前腋下,观察其致瘤能力;观察LM8/B7-1致敏的小鼠对骨肉瘤细胞LM8是否具有免疫保护作用。结果:B7-1基因在转基因细胞LM8/B7-1中能得到高表达;LM8/B7-1体外增殖能力与LM8和LM8/pcDNA3无明显差异(P>0.05);LM8/B7-1诱导的CTL的杀伤活力显著高于LM8和LM8/pcDNA3诱导的CTL对相同靶细胞的杀伤活力,LM8/B7-1诱导的CTL对LM8/B7-1的杀伤活力也明显高于对LM8和LM8/pcDNA3的杀伤活力(P<0.05);LM8/B7-1致瘤能力较LM8和LM8/pcDNA3细胞明显下降(P<0.01);LM8/B7-1致敏的小鼠对骨肉瘤细胞LM8具有免疫保护作用。结论:B7-1基因转染骨肉瘤细胞能诱导抗骨肉瘤主动免疫作用,为利用B7-1基因进行骨肉瘤免疫基因治疗提供了实验依据。 | 潘海涛 郑启新 杨述华 刘勇 叶树楠 | 2006 | 肿瘤2006，26，9： | 1 |
| 16 | 转染B7基因的U14疫苗对小鼠宫颈癌的防治作用研究显示文摘目的 :探讨转染B7基因的U14疫苗能否在中国 6 15系小鼠体内诱导抗宫颈癌主动免疫应答。方法 :将小鼠B7 1基因导入 6 15系小鼠宫颈癌U14细胞 ,建立高效表达B7 1的U14细胞株 (B7+U14 )及其细胞疫苗。 (1)用B7+U14疫苗免疫 6 15系小鼠 ,观察U14皮下移植成瘤情况 ;(2 )用B7+U14疫苗治疗 6 15系荷瘤小鼠。设立对照组 ,观察各组小鼠的成瘤情况、生存期 ;(3)体外实验检测经B7+U14和U14疫苗免疫小鼠T细胞杀伤肿瘤的活性。结果 :(1)B7+U14疫苗免疫可有效地预防野生型U14细胞移植瘤的产生 (比较肿瘤大小、生存期 :P <0 .0 1) ;(2 )B7+U14疫苗能治愈部分荷瘤小鼠 ,其效果受荷瘤大小的限制 ,肿瘤直径 >3mm时 ,B7+U14疫苗治疗无效 (P <0 .0 5 ) ;(3)经B7+U14和U 14疫苗免疫小鼠T细胞 ,其不同效靶比杀伤肿瘤效率前者明显高于后者 (P <0 .0 0 1)。结论 :转染B7基因的U14细胞疫苗能诱导机体产生有效的抗宫颈癌主动免疫应答。应用转染B7基因的肿瘤细胞疫苗可能成为临床术后治疗宫颈癌的有效辅助方法。 | 陶光实 胡锦跃 邹红卫 林秋华 刘凤英 吴宜林 孙去病 | 2003 | 现代妇产科进展2003，12，4： | 1 |
| 17 | DPC4RNAi慢病毒载体的构建及其基因沉默效应显示文摘目的 :构建DPC4基因RNA干扰慢病毒载体及其基因沉默效应。方法 :针对DPC4基因序列,利用网站设计程序按照RNA干扰序列设计原则,设计多个RNA干扰靶点序列,根据设计经验和设计软件进行评估测定,选择最佳的动力学参数靶点进入后续实验流程;生工生物合成含干扰序列的双链DNA oligo,其两端含酶切位点粘端,直接连入酶切后的RNA干扰载体上。将连接好的产物转入制备好的细菌感受态细胞,对长出的克隆进行酶切鉴定,挑选出阳性克隆测序,进行测序比对后,鉴定阳性克隆即为构建成功的目的基因RNA干扰慢病毒载体。结果:经过Western blot方法检测和测序证实,成功构建了DPC4 shRNA慢病毒载体LVshSmad4,并成功制备了DPC4shRNA慢病毒,3株病毒感染细胞后均具有有效的基因沉默,其中SH1序列最为显著。结论 :DPC4基因RNA干扰靶点的成功设计和RNA干扰靶点慢病毒载体制备,而且具有显著的基因沉默效果。 | 仲丽丽 白云 张英博 李洋 潘思文 刘宏 周忠光 | 2014 | 辽宁中医药大学学报2014，16，2： | 0 |
| 18 | 转染B7基因的U14疫苗对小鼠宫颈癌的防治作用研究(英文)显示文摘目的 探讨转染B7基因的U14疫苗能否在体内诱导中国 615系小鼠抗宫颈癌主动免疫应答。方法 将小鼠B7 1基因导入 615系小鼠宫颈癌U14细胞 ,建立高效表达B7 1的U14细胞株 (B7+U14)及其细胞疫苗。 ( 1)用B7+U14疫苗免疫 ,预防 615系小鼠皮下移植成瘤。 ( 2 )用B7+U14疫苗治疗 615系荷瘤小鼠。设立对照组 ,观察各组小鼠的成瘤情况、生存期。 ( 3)体外检测经B7+U14和U14疫苗免疫小鼠T细胞杀伤肿瘤实验。结果 将B7基因转染入U14细胞后获得了B7 1高表达克隆株 (B7+U14)。 ( 1)B7+U14疫苗免疫 ,可有效预防野生型U14细胞移植瘤产生 (比较肿瘤大小、生存期 :P <0 0 1)。 ( 2 )B7+U14疫苗能治愈部分荷瘤小鼠 ,其效果受荷瘤大小限制 ,即肿瘤直径 >3mm时 ,B7+U14疫苗治疗无效 (P <0 0 5)。 ( 3)体外检测经B7+U14和U14疫苗免疫小鼠 ,前者T细胞在不同效靶比杀伤肿瘤效率明显高于后者 (F =310 8,组间有显著差异P <0 0 0 1)。结论 转染B7基因的U14细胞疫苗能诱导机体有效的抗宫颈癌主动免疫应答。提示应用转染B7基因的肿瘤细胞疫苗可能成为临床术后治疗宫颈癌的有效辅助方法。 | 陶光实 胡锦跃 邹红卫 林秋华 刘凤英 吴宜林 孙去病 | 2001 | Chinese Medical Journal2001，，6： | 0 |
| 19 | 树突状细胞与肝癌细胞瘤苗的实验研究显示文摘目的 :探讨杂交瘤苗H2 2 DC的生物学特性及特异性CTL活性。方法 :利用Metrizamide等分离小鼠脾DC ,融合DC与H2 2 细胞 ,磁式细胞分选器分选出H2 2 DC ,从BALB/C小鼠皮下接种H2 2 DC法检测小鼠脾CTL活性。结果 :(1)H2 2 DC虽在体外能分裂增殖 ,但明显低于H2 2 细胞 ,无体内致瘤性 ;(2 )活H2 2 DC免疫的小鼠 ,其脾CTL杀伤H2 2 细胞的活性明显高于对照组 (H2 2 +DC、H2 2 、PBS)小鼠 (P <0 0 1)。结论 :DC H2 2 能诱导出特异性的CTL活性 。 | 张娟 张锦堃 陈海滨 | 2004 | 实用医学杂志2004，20，3： | 0 |
| 20 | HPV16 E6 DNA疫苗诱导小鼠抗宫颈癌主动免疫显示文摘目的 :探讨HPV16E6DNA疫苗能否在体内诱导小鼠抗宫颈癌主动免疫应答。方法 :将HPV16E6基因转染 6 15小鼠宫颈癌U14 ,建立高表达E6的U14细胞株即E6 + U14 ;制备HPV16E6DNA疫苗 ;随后分组进行体内预防和治疗E6 + U14移植同系小鼠试验 ,设立对照组 ;观察各组小鼠肿瘤大小 ,计算抑瘤率、生存时间 ;计算淋巴和肺转移率 ;体外分别检测经HPV16E6DNA疫苗免疫后的小鼠T淋巴细胞对E6 + U14和U14的杀伤作用。结果 :HPV16E6DNA疫苗预防和治疗组小鼠各时期瘤体大小均显著低于对照组 (均P <0 .0 0 1) ,两组抑瘤率均大于70 % ;平均生存时间均大于对照组 (均P <0 .0 0 1) ;预防组淋巴转移率低于对照组 (P <0 .0 5 ) ;HPV16E6DNA疫苗免疫小鼠T淋巴细胞在不同效靶比 ,对E6 + U14的杀伤效率均明显高于对U14者 (均P <0 .0 0 1)。结论 :HPV16E6DNA疫苗能诱导机体产生针对HPV16E6阳性的宫颈癌细胞主动免疫应答及特异性细胞毒淋巴细胞 。 | 陶光实 胡锦跃 刘凤英 吴宜林 刘毅智 李官成 | 2004 | 中南大学学报(医学版)2004，29，1： | 0 |
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