目的 探讨微小RNA-429(miR-429)过表达对乳腺癌细胞增殖及侵袭的影响,并分析其与Wnt信号通路的可能作用机制.方法 选取2017年8月至2018年6月手术治疗的76例乳腺癌患者,切除癌组织及癌旁组织作为乳腺癌组与癌旁组,采用实时定量聚合酶链反应(qRT-PCR)检测2组miR-429表达水平.体外培养乳腺癌细胞株MCF-7,采用脂质体Lipofectamin 2000介导的方法 转染细胞,分为空白对照组、阴性转染组以及miR-429 mimics组,倒置荧光显微镜下检测转染效率,qRT-PCR法检测3组细胞中miR-429表达水平;MTT法及平板克隆形成实验检测细胞增殖能力;Transwell法检测细胞侵袭能力;蛋白免疫印迹法(WB)检测ki67、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、细胞核β-链蛋白(β-catenin)及Wnt通路相关蛋白表达情况.结果 与癌旁组相比,乳腺癌组组织中miR-429表达水平明显降低(P<0.05);与空白对照组和阴性转染组相比,miR-429 mimics组的miR-429表达水平、细胞增殖抑制率显著升高,细胞克隆形成率、细胞侵袭数量及ki67、MMP-2、MMP-9、Wnt、GSK-3β、β-Catenin表达水平均显著降低,差异有统计学意义(P<0.05).结论 miR-429过表达可抑制乳腺癌细胞增殖及侵袭,其作用机制可能与抑制Wnt信号通路有关.

目的 探讨甲状腺癌SW579细胞中性别决定区Y框蛋白1(SOX1)表达及其抑制细胞增殖、侵袭的机制,并分析其与Wnt信号通路的作用关系.方法 体外培养正常永生化表皮细胞Hacat、甲状腺癌SW579细胞,采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)及蛋白免疫印迹法(western Blot,WB)检测SOX1 mRNA及蛋白表达水平.实验分为空白对照组(未经任何处理的SW579细胞);阴性转染组(SOX1阴性对照质粒的SW579细胞);SOX1过表达组(含有SOX1过表达载体重组质粒的SW579细胞);Wnt激活剂组(Wnt通路激活剂的SW579细胞);联合组(SOX1重组质粒转染后加入Wnt通路激活剂的SW579细胞).采用CCK-8法检测细胞增殖抑制率;Hoechst染色法检测细胞凋亡情况;Transwell法检测细胞迁移和侵袭;WB法检测β-连环蛋白(β-catenin)、糖原合成酶激酶-3β(GSK-3β)及其磷酸化蛋白(p-GSK-3β)、细胞周期相关蛋白1(CyclinD1)、c-Myc蛋白表达情况.结果 与正常组细胞比较,甲状腺癌SW579细胞中SOX1 mRNA及蛋白表达均显著降低(P<0.05);与空白对照组、阴性转染组、联合组比较,SOX1过表达组SW579细胞增殖抑制率、细胞凋亡率均显著升高(P<0.05),而Wnt激活剂组显著降低(P<0.05);与空白对照组、阴性转染组、联合组比较,SOX1过表达组SW579细胞迁移及侵袭数量、β-catenin、p-GSK-3β、CyclinD1、c-Myc蛋白表达水平均显著降低(P<0.05),而Wnt激活剂组显著升高(P<0.05).结论 SOX1可通过调控Wnt通路进而抑制甲状腺癌SW579细胞增殖及侵袭.

Objective:To explore the effects of subclinical hypothyroidism on hemorheology and levels of platelet-associated factors in patients with type 2 diabetes mellitus complicated with coronary heart disease.Methods:83 patients with type 2 diabetes mellitus complicated with coronary heart disease were included in the control group,and 83 subclinical hypothyroidism patients with type 2 diabetes mellitus complicated with coronary heart disease were included in the case group.Hemorheology,blood lipids and platelet-associated factors were compared.Results:The levels of TC,TG and LDL-C in case group were significantly higher than those in control group(P＜0.05).The level of plasma viscosity,as well as high cut and low cut blood viscosity in case group were also remarkably higher than those in control group(P＜0.05).In addition,in case group,the levels of PAF,GMP-140,β-TG,TXA2 were (124.1±37.5) μg/L,(5.8±2.6) × 1010/L,(41.0±13.7) μg/L,(407.3±48.5) ng/L,respectively,they were statistically increased compared with those in control group,which were (96.9±23.4) μg/L,(3.5±1.4)1010/L,(23.8±9.6)μg/L and (256.3±42.7) ng/L,respectively (P＜0.05).While,the level of PGI2 in case group (223.6±37.8)ng/L was significantly decreased as campared with that in control group (364.1±34.5)ng/L (P＜ 0.05).Conclusion:Subclinical hypothyroidism patients with type 2 diabetes mellitus complicated with coronary heart disease could cause increasing levels of TC,TG and LDL-C,and also induce increase of blood rheology and platelet-related factors.

Cyclin-dependent kinase (CDK) family members have been considered as attractive therapeutic targets for cancer. In this study, we aim to investigate the anticancer effects of a selective CDK7 inhibitor, BS-181, in gastric cancer (GC) cell line. Human GC cells (BGC823) were cultured with or without BS-181 at different concentrations for 24-72 hours. BS-181 significantly reduced the activity of CDK7 with downregulation of cyclin D1 and XIAP in GC cells. Treatment with BS-181 induced cell cycle arrest and apoptosis. The expression of Bax and caspase-3 was significantly increased, while Bcl-2 expression was decreased in cells treated with BS-181. In addition, the inhibition of CDK7 with BS-181 resulted in reduced rates of proliferation, migration, and invasion of gastric cells. Those results demonstrated the anticancer activities of selective CDK7 inhibitor BS-181 in BGC823 cells, suggesting that CDK7 may serve as a novel therapeutic target or the treatment of GC.

Objective To explore the relationship between gene mammalian STE20-like 1(MST1) expression and hepatitis B virus (HBV) infection and the hepatitis B virus x-protein (HBx) in hepatocellular carcinoma.Methods Collection of clinical specimens and cultured Hepatic carcinoma cell lines,By Western blotting and real-time quantitative polymerase chain reaction (Real-time PCR) method for detection of protein and mRNA levels and MST1 relationship with HBV and HBx.Extract genome of infection with HBV,Detect methylation of the promoter region.We tested the luciferase activity of MST1 promoter after adding different concentrations of methyl enzyme inhibitors 5-Aza-2'-deoxycytidine (5-Aza-CdR).Results MST1 was not deteded in 4 pairs liver tissue.The mRNA and protein espression levels were significantly lower than that of the adjacent tissues in 28 pairs liver tissue (P ＜ 0.05).The expression of MST1 in HBV infection was significantly lower than that of the control group (P ＜ 0.05).The rate of methylation of MST1 promoter region was 39％ after HBV infection,the rate of methylation of non HBV infected cells was 17％.And MST1 increases with the inhibitor concentration increasesafter adding 5-Aza-CdR (P ＜ 0.05).Conclusion Expression level of MST1 promoter in the cells containing HBV genome plasmid significantly decreased.

Human sulfatase-1 (hSulf-1) has been shown to desulfate cellular heparin sulfate proteoglycans and modulate several growth factors and cytokines. However, hSulf-1 has not been previously shown to mediate the signal transducer and activator of transcription 3 (stat3) signaling pathway, which is known to regulate cell proliferation, motility and apoptosis. The present study investigated the role of hSulf-1 in stat3 signaling in hepatocellular cancer. hSulf-1 expression vector and stat3 small interfering RNA (siRNA) were constructed to control the expression of hSulf-1 and stat3 in HepG2 cells. hSulf-1 was found to inhibit the phosphorylation of stat3 and downregulate its targeted protein. MTT and Transwell chamber assays, as well as Annexin V/propidium iodide double-staining methods, were used to examine the effects of hSulf-1 on stat3-mediated motility, proliferation and apoptosis in HepG2 cells. Transfection with hSulf-1 cDNA and/or stat3 siRNA inhibited cell proliferation and motility, concurrent with G0/G1 and G2/M phase cell cycle arrest and apoptosis. Overall, the results of the current study suggested that hSulf-1 functions as a negative regulator of proliferation and migration and as a positive regulator of apoptosis in hepatocellular carcinoma, at least partly via the downregulation of stat3 signaling.

Objective To observe the efficacy and safety of voriconazole in the treatment of malignant tumor patients with invasive fungal infection.Methods Forty four malignant tumor patients complicated with invasive fungal infection were randomly enrolled in two groups.They were administered by Voriconazole and Itraconazole respectively.Results In Voriconazole group,the cure rate and overall clinical efficacy rate were 68.2% and 77.3% respectively.The rate of side effects in Voriconazole group was 27.3%.In Itraconazole group,the cure rate and overall clinical efficacy rate were 61.9% and 76.2% respectively.The adverse reaction rate was 19.0%.But there were no significant differences in overall cure rate,clinical efficacy rate and adverse reaction rate between two groups.Conclusion Voriconazole was effective and safe in the treatment of malignant tumor complicated by invasive fungal infection,which is more economical than Itraconazole.

Objective To explore the molecular mechanisms of sodium butyrate(NaB)inducing apoptosis of human hepatoma cell line 7721.Methods The hepatoma cells were treated with various concentrations of NaB in different durations in vitro.Proliferation suppression was observed bv MTT method and invert microscope,and morphological changes of apoptosis were observed by transmission electron microseopy(TEM).Flow cytometry(FCM)was used to investigate the apoptosis rate and the cell cycle.The expression of p21 mRNA and protein was detected by semi-quantitative RT-PCR and Western-blot respectively.Results The NaB inhibited the growth of bepatoma cells in a dose-dependent and time-dependent manner.Invert microscopy and TEM showed that the cells treated wlth NaB exhibited characteristics of apoptosis.NaB blocked cells mainly in the G0/G1 phase,but stimulated p21 expression both at the mRNA and protein levels.Condusion NaB effect on proliferation inhibition and apoptosis induction may be linked to its ability to up-regulate of D21 gene.

Objective To explore the clinical characteristics of hepatic tuberculosis. Method To analyse 14 cases who were diagnosed hepatic tuberculosis by pathological examination.Result 64.28% of the 14 patients was diagnosed by percutaneous liver biopsy.Fever,hepatomegaly and abdominal pain were the major clinical manifestations,they were present in 71.43%,42.86% and 35.71% of the patients respectively.Anemia,liver function damage and increased ESR were noted in the patients (57.14%,57.14%,35.71% respectively).Conclusion The diagnosis should be considered in patients with unexplained fever associated especially with hepatomegaly or hepatosplenomegaly,liver function damage and increased ESR.Percutaneous liver biopsy is the most valuable method to confirm the diagnosis.

Objective To explore the clinical characteristics of hepatic tuberculosis.Methods The clinical data of 14 cases who were diagnosed as having hepatic tuberculosis by pathological examination were analyzed.Results 64.28% of the 14 patients was diagnosed by percutaneous liver biopsy. Fever,hepatomegaly and abdominal pain were the major clinical manifestations and present in 71.43%,42.86% and 35.71% of the patients respectively. Anemia,liver function damage and increased ESR were noted in the patients (57.14%,57.14%,35.71% respectively). Conclusion The diagnosis should be considered in patients with unexplained fever associated especially with hepatomegaly or hepatosplenomegaly,liver function damage and increased ESR.Percutaneous liver biopsy is the most valuable method to confirm the diagnosis.

Objective To investigate the effectiveness of splenectomy in patients with idiopathic thrombocytopenic purpura who failed to respond to conservative management.Method Fifty-eight patients were treated with splenectomy and the clinical outcome was observed.Results Platelet count recovered to normal (≥10×109/L) one week after operation in 45 cases(77.6%).The total effective rate was 75.6% when followed-up for 2 months, 80% when tbliowed-up for 6 months and 10 cases had recurrence.Conclusions Splenectomy is a safe and effective therapy for patients with ITP who failed to respond to conservative therapy and those who had an early increase of platelet count after operation have better prognosis.

将2.85 mmol/L丁酸钠(NaB)体外作用于人结肠癌SW480细胞12、24、48 h分别用半定量RT-PCR法与Western-blot法检测p21基因mRNA及蛋白表达变化;并用FCM检测细胞周期变化.结果 p21基因mRNA 及p21蛋白表达随时间延长逐渐增加;结肠癌SW480细胞周期阻滞于G0/G1期,S期比例明显减少,细胞增殖指数下降.认为NaB可诱导结肠癌细胞凋亡,其机制可能为细胞周期阻滞及p21蛋白高表达.

乙型病毒性肝炎(乙肝)肝硬化行胆道手术与择期门体分流手术相比,术后并发症多,病死率高.而胆道疾病并存肝硬化或其他慢性肝病临床常见,容易被忽视,如处理不当,可导致不良后果.我院近2年收治乙肝肝硬化并胆道疾病5例,均行手术治疗.现将诊治体会报告如下.

Objective:To explore the effect of tanreqing injection on acute pancreatitis (AP) with acute lung injury (ALI). Method: In 79 cases with AP, 53 cases accepted routine therapy of AP, and the orther 26 cases added tanreqing injection. Incidence of ALI and acute respiratory distress syndrome (ARDS) were observed, blood gas analysis and levers of IL-6, IL-8 and TNF-αin pe-ripherial blood were tested and recorded. Result:Incidence of ALI /ARDS and the levers of IL-6, IL-8 and TNF-a in tanreqing group were much lower than that in routine group (P <0. 05) , and PaO2 and PaO2/FiO2 were higher than the routine control group. Conclusion:Tanreqing injection can decrease the incidence of ALI and ARDS in patients with AP and improve hypoxemia.

丁酸钠（NaB）作为一种组蛋白去乙酰化酶（HDAC）抑制剂,已被证实能抑制多种体外培养的肿瘤细胞增殖、诱导肿瘤细胞衰老和凋亡。我们拟用NaB作用于人肝癌SMMC-7721细胞,采用FCM检测细胞周期、逆转录-聚合酶链反应（RT-PCR）方法及Weatern blot方法在mRNA及蛋白水平检测p21WAF。一、材料与方法1．细胞培养：人肝癌SMMC-7721细胞购自复旦大学肝癌研究所。细胞用含10%小牛血清的1640培养基(购自Gib-

器官移植后长期免疫抑制剂的应用所致的毒性作用以及供者的短缺,严重制约着器官移植的进一步发展.而采取基因治疗手段可调控移植物局部微环境的免疫反应,使移植物免遭排斥,也可诱导受者对供者抗原的特异性免疫耐受,使移植物存活时间延长,基因治疗手段还可培育转基因动物,用于异种器官移植,以缓解供者短缺的矛盾.

通过观察丁酸钠(NaB)对人肝癌SMMC-7721细胞抑制增殖和诱导凋亡作用,初步确定丁酸钠对体外培养人肝癌SMMC-7721细胞的最适作用浓度.将丁酸钠以不同终浓度、不同时间作用于人肝癌SMMC-7721细胞,用华罗庚优选法确定实验浓度,采用MTT比色法、倒置显微镜观察细胞增殖抑制;采用流式细胞术分析细胞凋亡率和细胞周期;电镜观察超微结构确定细胞凋亡情况.结果显示.丁酸钠对人肝癌SMMC-7721细胞生长的抑制呈剂量依赖和时间依赖,透射电镜观察到细胞皱缩、核质浓缩、核碎裂以及凋亡小体形成等凋亡特征的形态学改变.高浓度(≥4mmol/L)时,细胞在12h迅速出现坏死,电镜下观察,正常细胞膜结构消失,线粒体肿胀、破裂.流式细胞术分析细胞阻滞于细胞周期的G0/G1期,S期比例明显减少,细胞增殖指数明显下降.

目的 评价经圆韧带途径行Ⅲ段胆管空肠吻合术姑息性治疗不能切除的恶性梗阻性黄疸病人的临床效果。方法 对 1999年 1月至 2 0 0 2年 12月间 5例不能切除的肝门部恶性梗阻性黄疸病人实施经圆韧带途径显露Ⅲ段肝内胆管 ,并与空肠行Roux en Y式吻合术。对术后血清胆红素和碱性磷酸酶进行动态检测 ,并就病人住院时间、术后生活质量、生存时间进行观察。结果 5例病人实施肝内胆管空肠吻合术后血清胆红素逐渐降低至正常范围 ,2例病人临终前可维持无黄疸状态。术后 1例并发胆漏 ,1例住院期间因胃肠道恶性梗阻恶病质死亡 ,其他病人均存活超过 13个月 ,最长达 2 3个月。本组 5例术后均无胆管炎和复发性胆道梗阻。结论 经圆韧带途径肝内胆管空肠吻合术是恶性梗阻性黄疸病人姑息治疗的有效术式 ,损伤小 ,方法简便 ,减黄效果确切 ,病人生存质量提高

目的探讨重症急性胰腺炎(SAP)合并深部真菌感染的预防和治疗措施.方法将1998年7月～2002年6月收治的70例SAP病人随机分3组:大蒜素预防组、氟康唑(小剂量)预防组、对照组,比较各组的真菌感染发生率,治疗后真菌清除率及死亡率.结果大蒜素组真菌感染率明显低于对照组(16% ∶30%,P＜0.05)及氟康唑组(9% ∶30% ,P＜0.01).真菌感染发生后,采用治疗剂量的氟康唑和两性霉素B对大蒜素组、对照组的真菌感染病人有效,而对氟康唑组真菌感染病人无效.结论 预防性应用大蒜素、小剂量氟康唑均可明显降低SAP的深部真菌感染发生率.真菌感染发生后,如氟康唑治疗无效,应及时改用两性霉素B.

ObjectiveTo study the early diagnosis and prevention of fungal infection in severe acute pancreatitis(SAP). Method 1.SAP patients from July 1998 to June 2002 were prospectively randomized into 3 groups: garlicin prevention group, fluconazole (low dosage) prevention group and control group, the incidence of fungal infection in SAP was compared between the groups. For fungal infection patients, the fungal clearance and mortality rate were observed. 2.Clinical data of SAP patients with fungal infection and with simple bacterial infection was compared by multivariate logistic regression, and clinical characters and risk factors of fungal infection were evaluated. Results 1.There were lower incidences of fungal infection in garlicin group (16% vs. 30%,P0.05) and fluconazole group(9% vs. 30%,P0.01) than that in control group; After treatment with amphotericin B or therapy-dose fluconazole, the fungal clearance rate in garlicin group (3/4) and control group (4/7) was higher than that in fluconazole group (0/2); Mortality in the 3 groups were 3/7,1/4 and 2/2.2.There were 13 cases with fungal infection(FI) and 25 cases with bacterial infection(BI); Hospital stay (58 d vs. 43 d, P=0.04) and mortality(46% vs. 16%,P=0.05) of FI were significantly higher than that of BI; Risk factors of FI included diabetes, grade Ⅱ severity, multi-operation, intestinal and/or biliary fistulas. ConclusionsFungal infection is an independent risk factor of death in SAP. Prophylactic antifungal agents could reduce the incidence of fungal infection in SAP. Therapy-dose fluconazole could clear the fungus strain of most fungal infection cases.