目的:探讨睾酮在体外对颗粒细胞凋亡的诱导作用,并阐明其作用机制.方法:噻唑蓝(MTT)比色法检测人卵巢颗粒细胞SVOG的细胞增殖抑制率,筛选最佳药物作用浓度后,将SVOG 细胞分为对照组、睾酮组(1.0×10-5 mol·L-1睾酮)、睾酮+氟他胺组(1.0×10-5 mol·L-1睾酮+1.0×10-5 mol·L-1 氟他胺)、睾酮+牛磺熊去氧胆酸(TUDCA)组(1.0×10-5 mol·L-1 睾酮+1.5 g·L-1 TUDCA).采用实时荧光定量PCR(RT-qPCR)法检测各组 SVOG 细胞中剪切型X盒结合蛋白1(XBP1s)、激活转录因子4(ATF4)、转录因子C/EBP同源蛋白(CHOP)和死亡受体5(DR5)mRNA表达水平,流式细胞术检测各组 SVOG 细胞凋亡率.结果:与对照组比较,睾酮组SVOG细胞形态改变,异型细胞增多,胞质中出现较多颗粒,培养基中细胞碎片增多,SVOG细胞增殖抑制率升高(P<0.05);与睾酮组比较,睾酮+氟他胺组和睾酮+TUDCA组SVOG细胞形态变化减少,细胞增殖抑制率降低(P<0.05);流式细胞术,与对照组比较,睾酮组SVOG细胞凋亡率升高(P<0.05);与睾酮组比较,睾酮+氟他胺组和睾酮+TUDCA 组 SVOG 细胞凋亡率降低(P<0.05).RT-qPCR法,作用48 h后,与对照组比较,睾酮组SVOG细胞中XBP1s、ATF4、CHOP和DR5 mRNA表达水平均明显升高(P<0.05);与睾酮组比较,睾酮+氟他胺组和睾酮+TUDCA组SVOG细胞中XBP1s、ATF4、CHOP和DR5 mRNA表达水平降低(P<0.05).结论:睾酮可诱导人卵巢颗粒细胞凋亡并引起卵泡闭锁,其作用机制可能是雄激素与雄激素受体(AR)结合激活CHOP-DR5通路,诱导内质网应激(ERS)发生,促进颗粒细胞凋亡.

目的:探讨妊娠期糖尿病(GDM)对新生子代大鼠膈肌功能和氧化应激水平的影响,阐明其可能作用机制.方法:44只怀孕的8周龄SD雌性大鼠随机分为正常对照组和GDM模型组.妊娠第1天,GDM组大鼠单次腹腔注射35 mg·kg-1链脲佐菌素(STZ),建立GDM大鼠模型(建模成功18只);正常对照组大鼠单次腹腔注射相同体积的柠檬酸-柠檬酸钠缓冲液.分娩后,体外检测2组新生子代大鼠的膈肌收缩力,免疫荧光法检测子代大鼠膈肌纤维横截面积,常规HE染色观察子代大鼠膈肌病理形态表现,透射电镜下观察子代大鼠膈肌超微结构,采用试剂盒检测子代大鼠膈肌组织中丙二醛(MDA)和羰基化蛋白水平及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性.结果:体外膈肌收缩力检测,与正常对照组比较,GDM组子代大鼠膈肌最大紧张收缩力、最大强直收缩力和不同刺激频率下的膈肌收缩力均明显降低(P<0.05),膈肌疲劳指数明显升高(P<0.05).免疫荧光法检测,与正常对照组比较,GDM组子代大鼠膈肌组织中Ⅰ型和Ⅱ型膈肌纤维横截面积均明显缩小(P<0.05).HE染色,2组子代大鼠膈肌纤维结构完整,排列整齐、致密,膈肌细胞无明显肿胀,细胞核染色清晰.透射电镜下观察,正常对照组子代大鼠膈肌纤维排列整齐,明暗带及"Z"线规整且清晰可见,线粒体结构完整,嵴成隔板状,清晰可见;GDM组子代大鼠膈肌纤维明暗带及"Z"线无规则排列或消失,线粒体数量减少,更多线粒体出现凋亡呈空泡化.与正常对照组比较,GDM组子代大鼠膈肌组织中MDA和羰基化蛋白水平均明显升高(P<0.05),SOD和CAT活性明显降低(P<0.05).结论:GDM可引起子代大鼠膈肌收缩力降低、纤维横截面积减小和超微结构改变,而氧化应激水平的升高是影响其膈肌功能的原因之一.

目的:探讨子宫内膜腺癌组织中Tip60蛋白的表达与临床病理特征及预后之间的关系,初步阐明Tip60蛋白在子宫内膜腺癌发生发展过程中的作用.方法:选取子宫内膜腺癌患者84例,采用免疫组织化学SP法检测84例子宫内膜腺癌组织和36例正常子宫内膜组织中Tip60蛋白表达水平,分析其与患者临床病理特征之间的关系;采用Kaplan-Meier曲线分析Tip60蛋白的表达与患者预后的关系;采用Western blotting法检测人子宫内膜癌细胞株ISHIKAWA和HEC-1A中Tip60蛋白表达水平.结果:免疫组织化学检测,子宫内膜腺癌组织中Tip60蛋白阳性表达率为54.76％(46/84),明显低于正常子宫组织中Tip60蛋白阳性表达率[83.33％(30/36)](χ2=8.859,P=0.003).Tip60蛋白表达与患者年龄无关(χ2=1.073,P=0.300);FIGO分期(临床病理分期)中,Ⅰ期子宫内膜腺癌患者癌组织中Tip60蛋白阳性表达率明显高于Ⅱ期和Ⅲ-Ⅳ期患者(χ2=13.481,P=0.001);高分化子宫内膜腺癌患者癌组织中Tip60蛋白阳性表达率明显高于低分化和中分化癌组织的患者(χ2=8.962,P=0.013).子宫内膜腺癌患者的中位随访时间为58个月,Kaplan-Meier生存分析及Log-rank检验,Tip60蛋白高表达的患者生存率明显高于低表达患者(χ2=64.391,P=0.002).高表达雌激素受体(ER)的ISHIKAWA细胞中Tip60蛋白表达水平明显高于低表达ER的HEC-1A细胞(P<0.05).结论:Tip60蛋白在子宫内膜腺癌患者癌组织中和高表达ER的ISHIKAWA细胞中呈低表达,提示其参与子宫内膜腺癌的发生发展,检测组织中Tip60蛋白表达水平有助于子宫内膜腺癌的诊断及评估预后.

Objective:To investigate the synergistic killer effect of natural killer cells(NK cells) combined with tamoxifen(TAM) on breast cancer cells(BCC)through the experiment in vitro,and to explore its mechanism.Methods:Three kinds of BCC with different receptor expression levels were selected for the experiment.Blank control group, different concentrations of TAM groups and different time groups were set up.MTT assay was used to detect the inhibitory rates of proliferation of cells,and the final experiment concentration of 5 μmol·L-1 was determined.The cells were divided into natural-release group,largest-release group,TAM group,NK cells group, and combined-experimental group(BCC+NK cells+TAM),and the synergistic killer effect of NK cells combined with TAM in different effector-target ratios were detected with Calcein-AM release assay.In ELISA assay the cells were divided into blank control group (NK cells),NK cells+TAM group, NK cells+BCC group and combined-experimental group,and the levels of IFN-γ and TNF-α in the NK cells in various groups were measured.In flow cytometry detection the cells were divided into blank control group (NK cells),NK cells+TAM group,NK cells+ BCC group, and combined-experimental group,and the expression levels of NKp46,CD158a,CD158b,CD158b2,and CD158e were determined;while the cells were divided into blank control group (BCC),BCC+TAM group,BCC + NK cells group, and combined-experimental group,and the expression levels of the MICA,ULBP1 and ULBP2 were detected.Results: The MTT assay results showed that the inhibitory rates of proliferation of 3 kinds of BCC had obvious time-and concentration-dependence (P<0.05).The Calcein-AM release assay results showed that the killing-rates of BCC in TAM groups were increased with the increase effector-target ratios of compared with NK cells group;and the killing-rate in combined experimental group was obviously higher than those in NK cells and TAM groups(P<0.05).The ELISA assay results showed that the levels of TNF-α and IFN-γ of NK cells in various experimental groups with BCC or not were increased compared with blank control group(P<0.05 or P<0.01);the levels were significantly increased when combined with TAM (P<0.05).The flow cytometry results showed that the NKp46 expression levels in various experimental groups were elevated compared with blank control group(P<0.05);the expression levels of CD158a, CD158b,CD158b2, and CD158e were significantly decreased(P<0.05);the expression levels MICA,ULBP1, and ULBP2 in BCC were significantly increased (P<0.05).Conclusion:The NK cells combined with TAM has the synergistic killer effect on the BCC in vitro.The synergetic mechanism may be as follows: TAM could increase the secretion of TNF-α and IFN-γ of NK cells to enhance their cytotoxicity;TAM also could up-regulate the expression levels of activating receptors and activating ligands,and down-regulate the expression levels of inhibitory receptors to increase the killing ability of NK cells.

Objective:To discuss the expressions of hMOF protein and mRMA in endometrial cancer tissue,and to illuminate the possible mechanism of hMOF in the occurrence and development of endometrial cancer. Methods:The PT-PCR and immunohistochemistry methods were used to detect the expressions of hMOF protein and mRNA in 38 cases of endomerial cancer tissues and 20 cases of normal endometrial tissues,and to analyze the relationship between its expression level and the clinical characteristics and prognosis of the patients with endometrial cancer.Results:The immunohistochemistry results showed that the positive expression rate of hMOF protein in endometrial cancer tissue was lower than those in normal endometrial tissue (P=0.037);the positive expression rate of hMOF protein in stage Ⅰ patients was higher than those in stage Ⅱ and Ⅲ-Ⅳ patients (χ2=6.591,P=0.037);the positive exprssion rate of hMOF protein in the patients with poor differentiation was lower than those in the patients with moderate and high differentiation (χ2=7.351,P=0.025).The RT-PCR results showed that the percentage of the patients with expression up-regulation of hMOF mRNA in endometrial carcinoma tissue was remarkly decreased compared with normal endometrial tissue (P=0.003).Conclusion:hMOF protein may participate in the occurrence and development of endometrial cancer,and hMOF protein could be a biomarker for prognosis of endometrial cancer.

随着高新技术的深入,微创观念和微创外科发展迅速,宫腔镜作为微创的重要组成部分,改善了传统的妇科疾病,近30年来,宫腔镜技术在我国得到快速的发展和应用,它创伤小、治愈率高、手术和住院时间短、适用范围广,从最初的单纯宫腔镜检查术,到现在治疗各种妇科疾病的宫腔镜治疗,已被广大患者所认可,本文回顾性分析了吉大一院应用宫腔镜技术诊治542例妇科疾病患者临床资料、不同宫腔镜手术方法及预后。

甲状腺乳头状癌（PTC）是最常见的甲状腺恶性肿瘤，近年来发病率呈明显上升趋势。Ki-67是与细胞增殖相关的核抗原，鼠类肉瘤滤过性毒菌致癌同源体 B1（BRAF）基因所表达的蛋白参与细胞的增殖分化和凋亡，Ki-67及BRAF基因突变均可影响细胞周期的调控及肿瘤的发生发展。本文就Ki-67和 BRAF基因突变在PTC发生发展中的作用及其对肿瘤侵袭性、临床诊断及预后方面的影响进行综述。

Objective:To study the inhibitory effect of γδT cells on the proliferation of ovarian cancer SKOV3 cells,and to clarify its possible mechanism of inducing apoptosis. Methods:The human ovarian cancer SKOV3 cells cultured in vitro were used as control group,and theγδT and SKOV3 cells were co-cultured for 72 h as γδT cells treatment group.Laser scanning confocal microscope was used to obeserve the morphological changes of nucleus SKOV3 cells,and the inhibitory rate of proliferation of SKOV3 cells in two groups were detected by MTT method;Transwell Chambers was used to detect the cell migration ability,then the apoptotic rates of SKOV3 cells were tested by flow cytometry (FCM).Results:The apoptotic morphology of nucleus of SKOV3 cells in γδT cells treatment group were found under microscope,such as nuclear shrinkage.The MTT resultes displayed that the inhibitory rate of proliferation of SKOV3 cells in γδT cells treatment group was higher than that in control group (P <0.05).The Transwell Chambers results showed that the number of transmembrane cells in γδT cells treatment group was lower than that in control group,and the migration rate was decreased compared with control group (P <0.05).The FCM results showed that the apoptotic rate of SKOV3 cells in γδT cells treatment group was higher than that in control group (P < 0.05 ).Conclusion:γδT cells can inhibit the proliferation and the migration abilities of ovarian cancer SKOV3 cells,and promote the apoptosis.

Signal transducer and activator of transcription 3(STAT3) is a dual functional transcription factor with the functions of signal transduction and transcription regulation. It is reported that the expression of STAT3 in ovarian cancer is significantly higher and STAT3 can facilitate ovarian cancer growth and metastasis. To clarify the definite effect and molecular mechanism of STAT3 involved in ovarian cancer growth and metastasis, STAT3 expression was significantly downregulated by transfecting ovarian cancer model SK-OV-3 cells with the plasmid vector which express specific RNAi that targets human STAT3. The downregulated STAT3 not only decreased the invasion and migration but also inhibited the proliferation of SK-OV-3 cells. Western blot assay shows that the expression of vascular endothelial growth factor(VEGF) and that of Survivin were reduced in the cells with the plasma vector expressing specific RNAi that targets human STAT3. These results demonstrate that STAT3 involved in the invasion and migration of SK-OV-3 regulates the expression of VEGF and Survivin. In addition, VEGF and Survivin could play an important role in ovarian cancer growth and metastasis.

目的 探讨乙酰唑胺是否可以抑制体外培养的卵巢癌细胞的生长.方法卵巢癌细胞体外培养,调整细胞浓度为1 x 105/ml,分成实验组3瓶和对照组1瓶,实验组3瓶更换成乙酰唑胺浓度分别为1×10-8mol/L、5×10 -8moL/L、10×10-8mol/L的培养液,对照组培养液不变,分别收集24、48、72 h的细胞,用预冷的70％乙醇固定,调整细胞浓度1×106/ml,PI染色,进行流式细胞仪检测.每组重复3次.结果①与对照组比较,不同浓度实验组细胞凋亡率均增加,且随着药物浓度增加,细胞凋亡率也在增加,差异有显著性(P＜0.05).②各实验组间比较,差异均有显著性(P＜0.05).③随着时间增加,对照组和实验组所有的细胞凋亡率增加,24h与48 h之间,24h与72 h之间,48h与72 h之间,差异均有显著性(P＜0.05).结论乙酰唑胺对体外培养的卵巢癌细胞有明显的抑制作用,且与药物浓度和培养时间有关.

Objective To determine the expression of Aquaporin1(AQP1) in ovarian serous tumor and investigate it was relationship with ascites.Methods A total of 66 frozen ovarian tissues of serous tumor and 10 of normal ovarian tissues were examined for the expression of AQP1 by immunohistochemistry.Results ①It was statistically difference between either ovarian serous borderline tumor(5 case,62.5%) or ovarian serous malignant(38 case,82.6%) and normal ovarian tissue(2 case,20%)(P0.05).②Expression of AQP1 in ovarian serous malignant carcinoma has no relationship with it was histological grade(P0.05) and clinical stage(P0.05).③Expression of AQP1 was statistically different between the cases of with(26 case,72.2%)and without ascites(4 case,40%)(P0.05).Conclusion Expression of AQP1 in ovarian carcinoma was positive and might have relationship with ascites in ovarian carcinoma.

Oncostatin M (OSM), a pleiotropic cytokine, has been shown to have distinctive effects in different tissues. In ovarian carcinoma, it is commonly co-expressed together with its receptors but its precise role and the underlying molecular mechanism governing its activity remains unclear. The aim of the present study was to investigate the function of the recombinant human OSM (rH-OSM) in human ovarian cancer. The study demonstrated that rH-OSM promotes the proliferation of SKOV3 ovarian cancer cells. Western blot analysis showed that phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), phosphorylated-extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) and p-p38 protein levels increased in the cell lines treated with rH-OSM. Proliferation in SKOV3 cells induced by rH-OSM was suppressed by inhibitors of p-p38 or p-ERK1/2. Western blot analysis showed that p-STAT3 protein levels decreased in SKOV3 cells treated with inhibitors of p-p38 prior to treatment with rH-OSM. Also, p-STAT3 levels did not increase in cells treated with inhibitors of ERK1/2 prior to treatment with rH-OSM. Cell proliferation was moderately increased, and p-ERK1/2 and p-p38 protein expression were similarly affected in STAT3-RNAi knocked-down SKOV3 cells treated with rH-OSM compared to the control group. The data demonstrate that the growth-promoting activity of rH-OSM may be mediated through different signaling pathways. ERK1/2 and p38 proteins regulate STAT3 expression in SKOV3 cells, while STAT3 may be pivotal to the proliferation of SKOV3 cells in vitro.

Objective To analyze the risk factors of fevers after percutaneous nephrostolithotomy (PCNL) and to determine a more effective prophylaxis method.Methods A retrospective analysis of 320cases who underwent PCNL for renal calculi from 2008 to 2011 (men 233,women 87,age between 22 years to 72 years) was made.The average age of the patients and the average diameter of the stones were 42 years and 3cm(0.8 -6 cm),respectively.We analyzed factors such as age ( ＞60 years and ≤60 years),stone size ( ＞2.0 cm and ≤2.0 cm),operative time ( ＞60 min and ≤60 min),irrigation pump pressure ( ＞120 mm Hg and ≤ 120 mm Hg),obstructive conditions,preoperative urinary tract infection and fever.Wethen compared the fever rate with each of the risk factors.Results There were 59 cases with fever after PCNL; 18.4％ of the cases had a temperature over 38 ℃.There were two cases of pyemia.The patients whose stone diameter ＞ 2 cm had a post-operative fever rate of 22.4％.The fever rate in patients whose stone diameter ≤2.0 cm was 10.4％.The fever rate in patients with an operative time ＞60 min and ≤60 min was 27.2％ and 10.4％,respectively.Patients with irrigation pump pressure ＞ 120 mm Hg and ≤ 120mm Hg had post-operative fever rates of 28.3％ and 11.0％,respectively.There was statistical significance between each post-operative fever risk factor group.Conclusions The post PCNL fever risk factors are stone diameter ＞ 2.0 cm,operation time ≤60 min and irrigation pump pressure ＞ 120 mm Hg.The effective prophylaxis policy of post-operative fever are the pre-operative using of broad-spectrum antibacterial agents,shorter operative time and lower irrigation pump pressure.A two-stage operation procedure will also reduce the post-operative fever.

Apoptin is a chicken anemia virus-derived, p53-independent, bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells, but not of normal diploid cells. To explore the application of apoptin in tumor gene therapy, we used a recombinant fowlpox virus expressing apoptin protein (vFV-Apoptin) to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma(HeLa) cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide(MTT) assay, acridine orage/ethidium bromide(AO/EB) and annexin V staining test, respectively. The results show that vFV-Apoptin inhibites the proliferation of HeLa cells in vitro through inducing the apoptosis of HeLa cells, and the inhibition effect of vFV-Apoptin has a dose-effect and time-effect relationship. The results of animal models show that vFV-Apoptin significantly inhibits tumor growth, extends the lifespan of animals and improves the mean survival. Experimental results indicate that vFV-Apoptin has a potential application in the tumor gene therapy.

Objective To discuss the cause and management of delayed hemorrhage after minimally invasive percutaneous nephrolithotomy (PCNL). Methods From 2004 to 2009,there were 13 cases (10 men and 3 women, mean age 47 years) developed severe bleeding following PCNL. The cause of hemorrhage and treatments were retrospectively analyzed and summarized. Results The time of hemorrhage was 5-40 d. In 7 of 13 cases, the bleeding were was controlled by complete bed rest,hemostatic,balloon compression and clamped nephrostomy tube. Six of 13 underwent selective renal arteriography after ineffective with conservative treatment. The DSA showed 4 were false aneurysm and 2 were arteriovenous fistula. All the 6 cases were treated by the super-selective arteriolar embolization. The hematuria was disappeared 1- 3 d later. The intravenous pyelogram revealed the renal function kept well during follow-up visit. Conclusions Delayed hemorrhage is one of the severe complications, which may be caused when the renal puncture passage established as forming false aneurysm and arteriovenous fistula. The renal arteriography and super-selective arteriolar embolization could be a safe and effective treatment for the severe hemorrhage after PCNL.

The constitutive signal transducer and activator of transcription 3(STAT3)is high expressed in tumor specimens,and is related to tumor occurrence,metastasis and tumor-related drug-resistance.And researches suggests that P53,OSM,Piwi,ARHI,BRCA,VEGF,HIF-1α,survivin gene were increased in tumor samples which accompanied by abnormal expression of STAT3 phosphorylation,further studies found that P53,OSM,Piwi,ARHI,BRCA,VEGF,HIF-1α,survivin gene may directly or indirectly affect tumor occurrence and progress by activating STAT3.In addition,STAT3 plays an important role in tumor cells and host immune regulation.Activation of STAT3 transmitted from tumor cells to tumor immune cells,which provide favorable circumstances for the occurrence of tumor.The aim of the paper is to elucidate the function of STAT3 in cancer.It maybe a the threshold of controlling the tumor and a target for treating tumor.

China-Japan Union Hospital of Jilin University implemented the problem-based learning (PBL) in urologist clinical medicine teaching. This paper described the teaching methods used, analyzed the information feedbacked from the students and finally summarized the experience got from whole course.It increased of the students' interests and raised the student' s learning efficiency and cultivated the stadents' abilities at all aspects. It concluded that PBL should be promoted in the course of practice teaching for medical students.

Objective To compare the efficacy and safety of minimally invasive percutaneous nephrosomy for complicated renal calculi between holimium laser lithotripsy and ultrasonic lithotripsy.Methods 315 cases were analyzed including the average time of lithotripsy,the rate of stone completely expelled and the rate of complications.Results The average time of lithotripsy in ultrasound group(56 min)was lower than that in laser group(82 min)(P0.01);the clearnce rate of stone at stage Ⅰ operation in ultrasound group(81.5%)was higher than that in laser group(65.5%)(P0.01);the average hospitalization time in ultrasound group(6 d) was less than that in laser group(8 d)(P0.05);the incidence of hemoglobin decrease in ultrasound group(5.0%) was lower than that in laser group(12.7%)(P0.05).Conclusion Percutaneous nephrostomy with ultrasonic lithotripter for complicated renal calculi has the advautages of mini-invasion,less operative duration,radid recovery,safety and very effective and faster clearance compared with laser lithotripsy.

目前研究认为多种因素与宫颈癌的预后有关.该文主要从宫颈癌的病因学(高危型 HPV分布)、临床参数、病理参数、分子生物学指标等方面对宫颈癌的预后进行简单的概述.