目的:旨在探究miR-613在胶质瘤中的表达及对细胞增殖、侵袭和血管生成的影响.方法:根据细胞转染将实验分组为对照miRNA组(Control组)、miR-613模拟物组(mimics组)和miR-613 mimics+VEGFA组(VEGFA组).采用逆转录定量聚合酶链反应(RT-qPCR)检测胶质瘤细胞和组织中miR-613和VEGFA mRNA的表达水平;采用荧光素酶报告基因分析miR-613与血管内皮生长因子(VEGF)的关系;采用Western blotting检测VEGFA蛋白的表达水平;通过体外实验检测转染细胞的增殖能力、侵袭能力和管状形成能力.结果:与正常组织样本相比,胶质瘤Ⅰ-Ⅱ期组样本的肿瘤细胞呈现异形,具有深核染色,并且肿瘤细胞密度适度较低,而胶质瘤Ⅲ-Ⅳ期组样本的肿瘤细胞的核分裂活跃,具有明显的微血管增殖和明显的细胞异型性;miR-613在胶质瘤Ⅰ-Ⅳ期组织样本中显著降低(P＜0.05).在U87和U251细胞系的VEGFA-WT组中,与Control组相比,mimics组的荧光素酶活性显著降低(P＜0.05).与Control组相比,U87和U251细胞系中mimics组VEGFA的mRNA和蛋白表达水平均显著降低(P＜0.05).克隆形成实验、血管生成实验和细胞侵袭实验结果表明,与Control组相比,mimics组的克隆形成数量、细胞侵袭数、内皮细胞HUVEC的管状形成数和Ang-2蛋白表达水平均显著降低(P＜0.05);与mimics组相比,VEGFA组克隆形成数量、细胞侵袭数、内皮细胞HUVEC的管状形成数和Ang-2蛋白表达水平均显著升高(P＜0.05).结论:miR-613通过靶向VEGFA抑制了神经胶质瘤细胞的侵袭、增殖和血管生成,提示miR-613可能成为未来治疗胶质瘤的潜在靶点.

目的:通过网络药理学方法探讨独活镇痛的分子生物学机制.方法:独活的活性成分及其靶标基因通过TCMSP和CNKI数据库获取,疼痛相关靶标基因通过GeneCards和DisGeNET数据库获取.利用Cytoscape创建独活中化合物-靶基因和疼痛相关基因靶网络,利用STRING数据库分析关键靶标,利用DAVID和Bioconductor数据库分析途径富集.结果:网络分析确定了独活中10种化合物以及45个靶标基因与疼痛高度关联,蛋白互作网络中的核心基因是MAPK1、MAPK8、IL6、TNF、MAPK3、MMP9、CASP3、IL1B、PTGS2和CASP8,并确定了与疼痛相关的FoxO、NF-κB、MAPK、HIF-1等核心信号通路.结论:本研究结果提示可为进一步开发疼痛相关新药提供理论依据.

目的 探讨TRIM22在胶质瘤中的表达模式及功能.方法 收集30例胶质瘤患者的胶质瘤组织以及胶质瘤旁正常组织,并通过qRT-PCR和Western blotting检测TRIM22在胶质瘤组织和细胞系中的表达情况.为验证敲除TRIM22基因对U87细胞的增殖、迁移和侵袭的影响,将实验分为3组:Con-shRNA组、shRNA-TRIM22组和shRNA-TRIM22+LiCl组.Con-shRNA组细胞共转染pMSCV-TRIM22质粒和对照shRNA,shRNA-TRIM22组细胞共转染pMSCV-TRIM22质粒和sh-TRIM22,shRNA-TRIM22+LiCl组细胞共转染10μmol/L LiCl+pMSCV-TRIM22质粒和sh-TRIM22.通过CCK-8比色法测定细胞增殖;Transwell分析细胞的迁移和侵袭情况;Western blotting检测上皮-间充质转化过程(EMT过程)标志蛋白(E-cadherin和Vim-entin)和Wnt/β-catenin通路标志蛋白(cyclinD1和c-Myc蛋白)的表达水平.结果 与胶质瘤旁正常组织相比,胶质瘤患者组织样本中TRIM22蛋白和mRNA的表达水平均显著提高(P<0.01).与Con-shRNA组相比,shRNA-TRIM22组U87细胞中TRIM22的mRNA和蛋白表达水平均显著降低(P<0.01);EMT过程标志蛋白E-cadherin的表达水平显著升高(P<0.01),Vimentin蛋白表达水平显著降低(P<0.01);U87细胞的增殖、迁移和侵袭能力均显著降低(P<0.05).与Con-shRNA组相比,shRNA-TRIM22组U87细胞中cyclinD1和c-Myc蛋白的表达水平显著降低(P<0.01);与shRNA-TRIM22组相比,shRNA-TRIM22+LiCl组U87细胞中cyclinD1和c-Myc蛋白的表达水平显著升高(P<0.01),U87细胞的增殖、迁移和侵袭能力均显著升高(P<0.01).结论 TRIM22基因敲除抑制了U87细胞的增殖、迁移、侵袭和EMT过程,提示TRIM22在胶质瘤中可能是一个潜在的致癌基因.

目的 检测并分析前列腺特异性膜抗原(PSMA)与程序性细胞死亡蛋白1配体1(PD-L1)在原发性前列腺癌组织中的表达及相关性,探索PSMA/PD-L1联合检测与原发性前列腺癌患者临床病理特征及生存周期的相关性.方法 采用免疫组织化学染色法检测218例原发性前列腺癌组织中PSMA与PD-L1表达,应用Pearson相关分析前列腺癌组织中PSMA和PD-L1的免疫组织化学染色评分相关性.根据免疫组织化学染色法检测结果,将患者分为PSMA+ PD-L1+、PSMA+ PD-L1-、PSMA-PD-L1+、PSMA-PD-L1-四组,而后应用Spearman秩相关分析PSMA/PD-L1联合检测与原发性前列腺癌患者临床病理特征的相关性,应用Kaplan-Meier生存周期曲线分析PSMA/PD-L1联合检测与原发性前列腺癌患者术后生存周期的关系.结果 原发性前列腺癌组织中PSMA与PD-L1表达呈正相关,PSMA和PD-L1双阳性表达与前列腺癌患者临床分期和淋巴结转移呈正相关,与原发性前列腺癌患者术后5年总体生存率呈负相关.结论 PSMA与PD-L1在原发性前列腺癌组织中表达呈正相关,两分子联合检测有助于前列腺癌恶性程度评估及生存周期预测.

目的 探讨老年消化性溃疡患者的临床治疗和护理的疗效及价值.方法 选取2015年5月～2017年5月收治的32例老年消化性溃疡患者为研究对象,给予有效的治疗和护理,观察其疗效.结果 所有患者经治疗后,28例症状全部消失,治愈率为87.5％;而在治疗过程中出现恶心呕吐2例,胃酸1例,给予常规治疗后症状缓解,其中2例发生胃穿孔,需给予进一步的手术治疗.结论 对老年消化性溃疡患者给予有效的治疗和护理,具有良好的效果,可有效改善其治疗效果,降低不良反应发生率,提高患者的护理满意度,值得在临床上大力推广并应用.

目的:研究脂多糖(LPS)促进氧化低密度脂蛋白(ox-LDL)诱导的泡沫细胞形成的机制.方法:人源性THP-1细胞的培养,经ox-LDL诱导形成泡沫细胞.采用油红O染色鉴定泡沫细胞的形成,免疫荧光和Western blot方法检测自噬活性,观察自噬作用对泡沫细胞脂质沉积的影响.结果:①经形态学观察,脂多糖可以促进ox-LDL诱导的泡沫细胞的形成.②脂多糖可以激活自噬作用,并且自噬活性在16h达到最强.③脂多糖可以增强自噬激活剂雷帕霉素(Rap)的促自噬作用(P＜0.05),并且削弱自噬抑制剂3-甲基腺嘌呤(3-MA)的作用.④Rap单独作用不能影响泡沫细胞中脂质的累积,然而脂多糖能够增强Rap的作用,显著促进脂滴在泡沫细胞中的累积(P＜0.05);3-MA可以抑制基础水平和脂多糖诱导后泡沫细胞中脂滴的积累.结论:脂多糖通过增强自噬作用促进泡沫细胞的形成.

目的:观察银杏叶提取物对黑素细胞氧化应激损伤的影响.方法:用1.0mmol/L的H2O2处理人黑素细胞系PIG1细胞,采用CCK8法检测黑素细胞的活力,并于倒置显微镜下观察黑素细胞形态学的变化.结果:① 银杏叶提取物各浓度组对黑素细胞氧化应激损伤具有保护作用并呈剂量依赖性增强,与模型组相比差异均具有显著性(P≤0.05).② 倒置显微镜下观察可见H2O2组黑素细胞减少,细胞树突明显收缩,部分细胞变圆,不同浓度银杏叶提取物可明显抑制细胞树突收缩,并显示一定的剂量依赖性.结论:银杏叶提取物对氧化应激损伤的黑素细胞具有保护作用.

目的观察ROCK-Ⅰ选择性抑制剂Y-27632对体外培养的人眼Tenon囊成纤维细胞（ocular Tenon capsule fibroblasts,OTFs）增殖、凋亡和黏附性的影响。方法体外培养的第4～6代OTFs经溶血磷脂酸（lysophosphatidic acid,LPA）诱导后,不同浓度Y-27632处理,采用MTT测定细胞增殖抑制率和细胞黏附抑制率,未经LPA诱导OTFs细胞同时采用Annexin V-FITC/PI双标记流式细胞术测定细胞凋亡率,以及细胞平板克隆增殖实验测定不同浓度Y-27632组OTFs细胞增殖克隆形成率。结果 6、30、150、750μmol/L Y-27632各组经LPA诱导的OTFs细胞增殖抑制D（490）与LPA组比较,差异均有统计学意义（P＜0.05,P＜0.01）,OTFs细胞黏附抑制D（490）与LPA组比较,同样具有统计学差异（P＜0.05,P＜0.01）。随着Y-27632浓度增加,细胞凋亡率相应增加。未经LPA诱导的OTFs细胞克隆形成率随着Y27632浓度的增高而降低。结论 Y-27632有效抑制LPA诱导的OTFs增殖和细胞间黏附,诱导细胞早期凋亡。

A prospective review about the relationship between melanoma and nevus entitled "The evolution of melanoma diagnosis:25 years beyond the ABCDs" was published in "CA Cancer J Clin" on July 29,2010.We read it with great interests and raised some opinions that malignant melanoma incidence is closely related to nevus,some of skin cancer is resulted from nevus,and it is possible that pre-existing nevus can transform to melanoma.So nevus is an important factor in the prevention of melanoma.We should pay attention to the relationship between melanoma and nevus in clinical work.All these should be concerned with in the prevention and diagnosis of melanoma.Our suggestions were accepted by the editors and the letter we wrote was published in "CA Cancer J Clin" on August 30,2010.

AIM:To isolate and identify single chain Fv (scFv) antibodies against breast cancer from a constructed human phage display library.METHODS:Recombinant phages specific for breast cancer cells were enriched after four-round screening with MCF-7 cells. We selected the antigen-positive ones from the enriched clones by phage ELISA. The positive clones were used to infect E.coli HB2151 to express soluble scFv antibody. The antigen binding activity of the soluble antibodies was detected by Western blotting.RESULTS:The specific antibodies against MCF-7 cells were enriched after four rounds of affinity selection. SDS-PAGE and Western blotting showed a band at relative molecular mass 30 000 Da, which indicated soluble antibodies were present. ELISA analysis revealed that soluble antibodies had the affinity to a human breast cancer cell line MCF-7 but not to other cancer cell line, which demonstrated scFv could react specifically with breast cancer cells.CONCLUSION:We constructed a scFv phage library against human breast cancer with high capacity by phage display technology. The scfv was demonstrated successfully to be expressed in E.coli HB215 and have a specificity to breast cancer. Our findings may provide an alternative approach, and a basis for further studies on diagnosis and therapy of breast cancer.

白癜风是一种常见的色素障碍性皮肤病,是世界皮肤病的三大顽症之一.其临床特征主要表现为黑色素细胞减少或消失而出现色素脱失斑,严重影响患者的容貌、心理、工作和人际关系.本科室既往对42833例患者的流行病学调查显示,约64.3％的患者初发年龄小于20岁,给患者带来不同程度的个人生活和心理的困扰,并影响其生活、工作,婚姻等,部分心理素质差的患者甚至为此而萌发轻生的念头,也给家庭和社会带来了巨大的压力[1].因此,早期预防和有效治疗白癜风已受到广泛的关注.由于目前白癜风的发病机制尚不明晰,其治疗仍比较困难.笔者结合临床经验,认为"肝肾阴虚为本,肝郁血瘀为标"为白癜风的基本病机,提出治疗应"从肝肾论治"的治疗思路,浅述如下.

Objective To observe the acute and chronic toxical reactions of Shengxue Capsules on mice and rats to suggest the reference dosage for humans.Methods Shengxue Capsules were ig administrated to mice and rats twice a day,24 and 16 g/kg,respectively.The mortality rate and drug toxicity were observed and recorded.During long-term toxicological studies,rats were given capsules by intragastric administration at the dosage of 0.5,2.5,5.0 g/kg,respectively,twice a day for 26 weeks.Hematology,blood biochemistry,measure weights and weight indexes,and pathological changes in major organs were examined at 13th week,26th week and 4 weeks after drug withdrawal.Results The maximum ig administration dosage was greater than 24 g/kg in mice and 16 g/kg in rats,240 and 160 times the clinical dosage in humans.Hematology,blood biochemistry,weights and weight indexex of rats were proved to be in the normal range and no pathologic changes in major organs were found.Conclusion Shengxue Capsules are safe at the prescribed dose.

Objective To discuss and establish the method for mass culturing dendritic cells from human peripherial blood.Methods The following five steps for mass culturing dendritic cells from human peripherial blood are established:①To use Ficoll discontinuous density gradient centrifugation to separate PBMC;②To use RPMI 1640 culture medium for centrifugalization and removing thrombocytes;③To use the ephemeral adhesion of DC precursors to remove suspending cells and strongly adhesive cells;④DC were isolated from human peripheral blood with the induction of mononuelear cells by recombinant human granulocyte/macrophage colony stimulating factor and recombinant human interleukin 4.⑤Mature dendritic cells were induced by recombinant human tumor necrosis factor.The expression of surface molecules of DC was examined by fluorescent microscopy and scanning electron microscopy for morphological study.Results Nine days after induction,the expression of CD40,HLA-DR and CD80 had been observed under fluorescent microscopy.The morphological appearance of DC was irregular under SEM.Numerous dendrites radially protrude from cell bodies,some of them being as long as 2 or 3 times of cell body,and the dendrites cross and fuse with each other.A large quantity of circular or oval small dendrites exist at the base of long dendrites,they were in conformity with the morphology of DC as reported before.Conclusion Highly effective and typical DC were obtained by this method,and this method is stable,simple,economic,and it can be used for clinical immunotherapy.

目的:建立放射性损伤的小鼠血虚证模型,观察三康胶囊的药理作用.方法:选取昆明种小鼠随机分为正常对照组、模型组、阳性对照组及三康胶囊低、中、高剂量组.采用60C-γ射线照射的方法造成小鼠血虚证模型,分别观察给药后第14、21、28、60天外周血象以及骨髓细胞形态学改变,以此考察三康胶囊的疗效.结果:①低、中、高剂量三康胶囊给药后第14、21、28、60天对放射性损伤的小鼠血虚证所致的红细胞(RBC)、白细胞(WBC)、血小板(PLT)和血红蛋白(HB)降低均具有显著升高作用(P＜0.05或P＜0.01);②60天镜下观察可见:正常对照组小鼠骨髓增生极度活跃,巨核细胞、粒细胞、红细胞大小、形态正常;模型组有8例小鼠骨髓增生不良,退化细胞胞浆不完整,胞浆胞核溶解,涂片中有空泡;其余4例骨髓增生极度活跃,有大量的粒细胞(以中性杆状粒细胞、中性分叶核细胞为主),幼稚红细胞;各剂量给药组小鼠骨髓增生极度活跃,有粒细胞、晚幼粒细胞、中性杆状粒细胞、中性分叶核细胞及幼稚红细胞逐步增多;三康胶囊低、中、高剂量组分别有6、4、3例小鼠骨髓增生不良退化,细胞胞浆、胞核不完整逐步减少.阳性对照组小鼠骨髓增生极度活跃,有大量的粒细胞(以中性杆状粒细胞、中性分叶核细胞为主),幼稚红细胞;其中有3例小鼠骨髓增生不良,退化细胞胞浆、胞核不完整.结论:三康胶囊对放射性损伤的小鼠血虚证模型有一定的治疗作用.

Objective: To investigate the suppressive effect of Prunella vulgaris on the growth and the proliferation cycle and apoptosis of human thyroid cancer cell line SW57.Methods: The survival rate and the inhibitory rate of SW579 cell in vitro were detected by MTT colorimetrie assay and cell growth curve assay at different time points under different concentration of Prunella vulgaris;The cell proliferation cycle and the apoptotic rate were examined by flow cytometryanalysis.Results: Prunella vulgaris inhibited the prolifera-tion of SW579 cell at G0/Gl phase,decreased the cell ratio at S phase.It suppressed the growth of tumor cells and raised the apoptotic rate in a concentration and time depending manner in a certain extent.Conclusion: Prunella vulgaris could suppress the proliferation of SW579 cell to induce apoptosis and block cell cycle.

目的:观察酸奶对大鼠胃黏膜损伤的促进或抑制修复作用.方法:利血平0.5 mL/kg腹腔注射,制作大鼠胃溃疡模型,然后各组大鼠分别用酸奶,纯奶,生理盐水5 mL/kg每天1次灌胃,7 d后,处死并解剖大鼠.观察胃黏膜损伤情况、测定胃黏膜中前列腺素E2(PGE2)的表达量、超氧化物歧化酶(SOD)表达量来评估酸奶对胃溃疡的损伤或修复作用.结果:3组大鼠胃黏膜均有点状出血,Guth积分法得出酸奶组较纯奶组积分少,有统计学差异(1.33±0.577 vs 5.00±1.323,P<0.05),镜下病理切片观察发现,各组大鼠胃黏膜不同程度充血,生理盐水组上皮严重脱落且腺体萎缩,酸奶组损伤不明显,仅有少量中性粒细胞浸润和上皮脱落.酸奶组PGE2表达明显比生理盐水组表达量高,有统计学差异(248.6±43.73 vs79.52±31.74,P<0.05).酸奶组SOD表达量比纯奶组低,差异有统计学意义(148.1±2.18 vs156.6±1.66,P<0.01).结论:酸奶对大鼠胃溃疡的恢复有一定的促进作用,同时胃黏膜PGE2表达量升高,SOD表达量下降.

Objective: To explore the relationship between serum estradiol (E2) and expression of parathyroid hormone receptor 1 (PTHR1) in ovariectomized osteoporosis Rats. Methods: Twenty adult female rats were randomly divided into two groups: the ovariectomized group and the model control group. The rats of castration group excised double ovaries and induced hormonoprivia. After three months, gained blood from cardinal vein and detected the level of serum E2; put all rats to death and got femur bone tissue with asepsis condition. Detected the transcriptional level of PTHR1 gene with RT-PCR. And detected the expression level of PTHR1 protein with Western blot. Results: In ovariectomized groupthe level of serum E2 was 13.80±1.10 pmol/L,the E2 of model control group was 23.51±1.20 pmol/L. In ovariectomized group right femur skeleton strength: fracture test was 9.89±0.65 N/kg, crushing test was 16.10±1.23 N/kg and bone density was 0.0730±0.0075 gms/cm2 . In control group, right femur skeleton strength: fracture test was 11.74±0.95 N/kg,crushing test was 19.38±1.84N/kg and bone density was 0.0839±0.0097gms/cm2. All indexes in ovariectomized group compared with model control group were obviously lower. The level of PTHR1 gene and protein in ovariectomized were group compared with model control were group obviously lower. Conclusions: Castration could lower the level of serum E2 obviously and induce osteoporosis in rats. It is very interesting that the level of PTHR1 gene and protein are obviously lower. It implicated that PTHR1 might play an important role in ovariectomized rats to induce osteoporosis.

中医学认为,肝肾阴虚、肝阳上亢是高血压病的主要发病机理,平肝潜阳法是高血压病最常用的临床治则.随着高血压病中医辨证分型客观化研究的不断发展,对平肝潜阳法的作用机制也从整体、器官、细胞、分子等水平进行了初步探讨,取得了一定的成效.本文就近10年来平肝潜阳法治疗高血压病的作用机制研究进展作一综述.

Objective To investigate the role of CD147 in glioma invasion, metastasis, and angiogenesis. Methods Glioma celt fine U251 was transfectzd with a CD147 antisense RNA expression vector. After selection and identification of the transfected cells, gelatin zymography, scrape-wound migration assay and Matrigel Boyden chamber assay were performed to analyze the inhibitory effects on the invasiveness and angiogenesis of the glioblastoma cells. Enzyme-linked immunosorbent assay (ELISA) was used to detect vascular endothelial growth factor (VEGF) level in the supernatant of the transfected cells. Result Gelatin zymography showed that MMP-9 and MMP-2 production was decreased after antisense RNA transfection of the cells in comparison with the blank control and empty vector-transfected U251 cells. Matrigel Boyden chamber assay demonstrated a significant reduction in the invasiveness of the transfected cells. Conclusion CD147 antisense RNA can inhibit the invasion, metastasis, and angiogenesis of glioma cells in vitro, suggesting the potential of CD147 as a new target for tumor therapy.

癫痫是神经系统常见疾病,发病率为1%,全世界约5 000万患者,我国目前约有900万患者.约60%～70%患者通过药物可以控制,30%以上属难治性癫痫,约7%～8%可进行外科治疗,还有大约25%患者目前无有效治疗,因此,从发病机制着手研究探寻新的有效治疗手段仍是当今癫痫研究的重点.