Objective: To explore the effect of TRAF6 gene silencing on the function of hepatocellular carcinoma SMCC7721 and its possible mechanism. Method: Cell lines were constructed by cell transfection technology and verified by quantitative real-time PCR. Cell functional changes were observed by CCK8 method, Transwell test and Method of EdU. Western blotting was used to explore the possible mechanism of action. Result: TRAF6 RNA was abnormally up-regulated in HCC, and TRAF6 levels were detected in both HCC cell lines and L02 cells. SMCC7721 was selected as TRAF6 high expression cell. The results of CCK8 assay and EdU method showed that the decrease of TRAF6 expression significantly inhibited the proliferation of SMCC7721 cells. The results of CCK8 assay and EdU method showed that the decrease of TRAF6 expression significantly inhibited the proliferation of SMCC7721 cells. Overexpression of TRAF6 in TRAF6 knockdown cells can restore and enhance cell proliferation. Transwell assay confirmed that the invasiveness of SMCC 7721 cells treated with siRNA was significantly reduced. After treatment with LV-Rescue plasmid, the cell invasion was restored and enhanced. Western blotting showed that the protein levels of YB-1, Wnt, β-catenin, c-myc and Cyclin D1 were significantly down-regulated in siRNA group. On the contrary, the expression level of CYLD protein increased. Conclusion: As an important intracellular junctive protein in tumor cells, TRAF6 may improve the expression of pro-cancer factors C-myc and Cyclin D1 by modifying (ubiquitination) YB-1, thus improving the proliferation ability of cells. This process may be closely positively correlated with the Wnt/β-catenin pathway, and negatively correlated with the expression of CYLD protein.

目的:探索LINC AC004463.6通过PI3K/AKT/BCL-2通路对肝细胞癌(hepatocellular carcinoma,HCC)细胞生物学行为的影响.方法:运用细胞转染技术构建细胞系,实时荧光定量PCR验证,运用CCK8法、Transwell实验、EdU实验和流式细胞仪检测凋亡技术观察细胞功能学变化,运用Western blotting和ELISA技术探索其可能作用机制.结果:LINC AC004463.6在HCC中异常上调,在HCC细胞系和L02细胞中均检测到LINC AC004463.6的水平.选取SMCC7721和QGY7701作为LINC AC004463.6高表达细胞.CCK8和EdU实验结果表明,LINC AC004463.6表达降低显著抑制了SMCC7721和QGY7701细胞的增殖.而LINC AC004463.6基因敲除细胞中过表达LINC AC004463.6可使细胞增殖恢复和增强.Transwell实验检测表明shRNA处理的SMCC7721和QGY7701细胞侵袭性显著降低;经Lv-Rescue质粒处理后,细胞侵袭恢复并增强.LINC AC004463.6表达降低的细胞凋亡水平较高;LINC AC004463.6基因在LINC AC004463.6基因敲低的细胞中过表达后,细胞的凋亡水平有所恢复.Western blotting和ELISA检测提示shRNA组细胞BCL-2蛋白表达水平明显下调,同时PI3 K和AKT蛋白表达水平明显下降;与此相反BAX蛋白表达水平升高.结论:LINC AC004463.6可能通过PI3K/AKT/BCL-2通路改变肿瘤细胞的生物学行为.LINC AC004463.6可能是诊断HCC的一个前瞻性生物标志物.

目的 探究转录因子7-like 2(transcription factor 7-like 2,TCF7L2)对原发性肝癌大鼠Th1/Th2免疫漂移及微血管密度(microvessel density,MVD)形成的影响.方法 60只Wistar大鼠根据随机数字表法分为空白对照组、模型组、si-NC组和si-TCF7L2组,每组15只.除空白对照组外,其余3组大鼠通过饮用二乙基亚硝胺(DEN)制备原发性肝癌模型.14周后,si-NC组大鼠尾静脉注射阴性对照慢病毒混合液,si-TCF7L2组大鼠尾静脉注射LV-TCF7L2-siRNA慢病毒混合液,空白对照组和模型组大鼠注射等体积生理盐水,每日1次,连续7 d.6周后,大鼠腹主动脉采血,全自动血液分析仪检测血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)及总胆红素(TBIL)含量;ELISA法检测干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、IL-4和IL-10含量;HE染色观察大鼠肝组织形态学变化;免疫荧光染色检测大鼠肝组织血管内皮细胞生长因子(VEGF)表达;免疫组织化学染色检测大鼠肝组织INF-γ、IL-4表达及MVD形成情况;实时荧光定量PCR检测肝组织中转录因子T-bet和Gata-3 mRNA表达;流式细胞术检测脾脏淋巴细胞Th1、Th2比例及Th1/Th2比值.结果 与空白对照组比较,模型组大鼠血清中ALT、AST、TBIL升高(P＜0.01),IL-2、IFN-γ水平下降,IL-4、IL-10水平升高,IFN-γ/IL-4比例降低(P＜0.01),肝组织出现肿瘤细胞,排列紊乱且核质变大、核仁突出,VEGF和IL-4表达水平变高,IFN-γ表达水平降低(P＜0.01),MVD增加(P＜0.01),T-bet mRNA相对表达量下调,Gata-3 mRNA相对表达量上调(P＜0.01),Th1细胞比例减少,Th2细胞比例增加,Th1/Th2比值降低(P＜0.01).与模型组比较,si-TCF7L2组大鼠血清中ALT、AST、TBIL降低(P＜0.01),IL-2、IFN-γ水平升高,IL-4、IL-10水平下降,IFN-γ/IL-4比例升高(P＜0.01),肝组织癌变程度减小,VEGF和IL-4表达水平降低,IFN-γ表达水平升高(P＜0.01),MVD减少(P＜0.01),T-bet mRNA相对表达量上调,Gata-3 mRNA相对表达量下调(P＜0.01);同时,Th1细胞比例增加而Th2细胞比例减少,Th1/Th2比值升高(P＜0.01).结论 沉默TCF7L2表达可效抑制原发性肝癌大鼠模型瘤体中微血管密度,调节Th1/Th2比例趋于正常.

Objective:To investigate the expression of microRNA (miR)-938, its effect on cell proliferation and its regulatory mechanism in hepatocellular carcinoma (HCC).Methods:HCC and paracancerous tissues were collected from 40 patients with HCC who underwent surgical resection in the Second Affiliated Hospital of Nantong University from January 2015 to June 2019, including 25 males and 15 females, with an average age of 61.4 years. HepG2 cells in the miR-938 overexpression group were transfected with miR-938 mimics, and the negative control group was transfected with the negative control sequence. Cell proliferation was detected by kit, the expression of miR-938 and the succinate dehydrogenase complex subunit D (SDHD) was detected by fluorescence quantitative polymerase chain reaction, and SDHD protein expression was detected by Western blot. The target genes of miR-938 were verified by dual luciferase reporting.Results:The relative expression of miR-938 in HCC tissues was (0.060±0.002), which was higher than that in adjacent tissues (0.030±0.002), and the difference was statistically significant ( P<0.05). The mRNA relative expression of SDHD in HCC tissues was (0.028±0.002), lower than that in adjacent tissues (0.062±0.002), and the protein expression of SDHD in HCC tissues was (0.963±0.008), lower than that in adjacent tissues (1.083±0.037), with statistical significance (both P<0.05). The proliferation activity of miR-938 overexpression group was significantly higher than that of negative control group, and the difference was statistically significant ( P<0.05). MiR-938 significantly inhibited the luciferase activity of SDHD wild-type 3’-untranslated regions. In the overexpression miR-938 cells, SDHD mRNA and protein levels were significantly lower than those in the negative control group ( P<0.05). Conclusion:MiR-938 was highly expressed in HCC tissues. MiR-938 promoted the proliferation of HCC cells by inhibiting the expression of SDHD.

目的 为探究JAK2/STAT3信号通路与炎性因子白细胞介素6、1β、10(interleukin-6、1β、10)的关系,IL-6、IL-1β、IL-10是否参与刀豆蛋白A(ConA)诱导的小鼠肝纤维化过程以及川芎嗪对其的影响.方法 BALB/C小鼠随机分为5组,正常对照组(A 组)10只;ConA 模型(B组)15只;川芎嗪低剂量(100 mg/kg)组(C组)15只;川芎嗪中剂量(200 mg/kg)组(D组)15只,川芎嗪高剂量(800 mg/kg)组(E组)15只.通过实时荧光定量 PCR 检测 IL-6mRNA、IL-1βmRNA和 IL-10mRNA 的表达水平;Western-blot检测 JAK2、STAT3和 p-JAK2、p-STAT3的表达情况.结果 与模型组相比,小鼠经不同剂量川芎嗪干预8周后,IL-6mRNA 和IL-1βmRNA 表达水平均降低,且与给药剂量呈相关性;IL-10mRNA表达水平升高,且与给药剂量呈相关性;p-JAK2、p-STAT3蛋白表达均降低,且下降程度与给药呈剂量相关性;JAK2和 STAT3蛋白表达变化不明显.结论 川芎嗪可能通过JAK2/STAT3信号通路及炎性因子 IL-6、IL-1β、IL-10对ConA诱导的小鼠肝纤维化产生保护作用.

The occurrence of Lung Cancer is the interaction of internal and external factors,mostly due to lung deficiency,lung Yin deficiency and the invasion of six Evils into the Lung.Commonly used Chinese medicine types are Heat-clearing,Mass-resolving,Phlegm-dispelling,Spleen-tonifying and other invigorating types,in which Heat-clearing type of Chinese medicine is the main type in treatment of lung tumor and can be used at all stagcs of the disease,particularly in the early stage of Lung Cancer.These medicines include Scutellaria Barbata,Houttuynia,Houttuynia,Fagopyrum and so on.At the same time the Mass-resolving and Phlegmdispelling types of traditional Chinese medicine can be supplemented to enhance the effectiveness of dispelling excessive evils.The commonly used medicines are Ginger Pinellia,Fritillaria,Campanulaceae,Rhizoma Amorphophalli,Rattus norvegicus and so on.Besides,the Spleen-tonifying and other invigorating types of Chinese medicines are commonly used at the late stage of Lung Cancer,which can play a positive effect on patients' constitution building.These medicines include Astragalus,Codonopsis,Poria,Atractylodes,Licorice and so on.

分析青霉素的中药药性理论,认为青霉素属寒凉之性,味苦,归心、肺、脾、肝、肾诸经;根据“取象比类”理论研究,可归于“清热药”之属;根据用量、归经的不同,轻则清热解表,重则解毒排脓,亦有祛风除湿的“风药”特性.基于此,青霉素作为“清热药”可适用于多系统、多病症,是一味药性复杂、功效丰富的“中药”.

目的分析2007-2009年我院药物严重不良反应发生的情况,以促进临床安全、合理用药。方法回顾性调查我院2007-2009年45例严重不良反应病例报告,从患者的年龄、性别、药品种类、给药途径等方面进行统计和分析。结果 45例严重不良反应中,性别无显著差异,以≥51岁老年患者居多;经静脉给药引发者39例占86.67%;所用药物中抗肿瘤药居首位,代表药物为紫杉醇和多西他赛;主要的临床表现为疾病、全身性损害。结论继续加强临床药物严重不良反应的监测,为临床安全合理用药提供依据。