目的:探讨MDT教学模式相比传统教学模式在妇科肿瘤学研究生临床教学中的应用效果.方法:将于2016年9月—2017年9月在哈尔滨医科大学附属肿瘤医院妇科进行临床学习的40名研究生随机分为两组,观察组采用MDT教学模式,对照组采用传统教学模式,比较两组研究生的临床教学成绩、教学满意率.结果:观察组的理论知识成绩、临床实践能力明显高于对照组(P<0.05);观察组在教学模式、对疾病分析逻辑思维能力、自主学习能力、医患沟通能力、学习参与感等满意度评分上明显高于对照组(P<0.05).结论:MDT教学模式在妇科肿瘤学研究生临床教学中的应用效果显著,能有效提高教学质量,提高研究生的综合临床素质,有利于为肿瘤科临床培养高素质的专业人才.

BACKGROUND:Activation of autophagy flux contributed to resistance of breast cancer (BC) cells to current chemotherapeutic drugs, which seriously limited their therapeutic efficacy and facilitated BC recurrence in clinic. However, the detailed mechanisms are still not fully understood. In the present study, we identified that inactivation of AMPK-ULK1 signaling cascade mediated protective autophagy sensitized BC cells to doxorubicin in vitro. METHODS:Cell counting kit-8 (CCK-8) assay and colony formation assay were performed to evaluate cell proliferation abilities. Trypan blue staining assay was used to examine cell viability, and Annexin V-FITC/PI double staining method was conducted to determine cell apoptosis. The autophagosomes in BC cells were observed and photographed by electronic microscope (EM). Western Blot analysis was employed to examine genes expressions at protein levels. RESULTS:The parental doxorubicin-sensitive BC (DS-BC) cells were exposed to increasing concentrations of doxorubicin to establish doxorubicin-resistant BC (DR-BC) cells, and the DR-BC cells were much more resistant to high-dose doxorubicin treatment compared to the DS-BC cells. Interestingly, high-dose doxorubicin specifically increased LC3B-II/I ratio, promoted autophagosomes formation and decreased p62 expression levels to facilitate autophagy in DR-BC cells, instead of DS-BC cells, and the autophagy inhibitor 3-methyladenine (3-MA) enhanced the cytotoxic effects of high-dose doxorubicin on DR-BC cells. In addition, we proved that high-dose doxorubicin triggered protective autophagy in DR-BC cells by activating AMPK-ULK1 pathway. Functionally, high-dose doxorubicin increased the expression levels of phosphorylated AMPK (p-AMPK) and ULK1 (p-ULK1) to activate AMPK-ULK1 pathway in DR-BC cells, and the inhibitors for AMPK (compound C) and ULK1 (SBI-0206965) blocked autophagy to promote cell death and slow down cell growth in DR-BC cells treated with high-dose doxorubicin. CONCLUSIONS:Collectively, our in vitro data indicated that blockage of AMPK-ULK1 signaling cascade mediated protective autophagy might be a promising strategy to increase doxorubicin sensitivity for BC treatment.

生物信息技术的快速发展正在推动一些隐藏的基因被人们所发现.环状RNA(circRNA)是内源性非编码RNA(ncRNA)家族的新成员,通过单链RNA分子的3’端和5’端连接而成,具有一定的广泛性、保守性、稳定性和组织特异性.近年来,随着高通量测序技术和生物信息学的不断发展,大量内源性circRNA在各种细胞类型中均可被检测到,且circRNA具有发挥微小RNA(miRNA)海绵作用、调控基因表达及翻译等生物学功能,同时在生物发育过程中和疾病诊断中起着至关重要的作用.circRNA具有差异性表达的特点,与多种恶性肿瘤的发生、发展密切相关,在肿瘤中发挥抑癌基因或癌基因的作用,有望成为恶性肿瘤的新型诊断标志物、分子治疗靶点和预后评估指标.目前,仅有少量关于circRNA在卵巢癌中研究进展的报道,本文拟从circRNA的生物学功能及其在卵巢癌中的研究进展进行简要综述.

目的:探讨三维动脉自旋标记(3D-ASL)与动态对比增强磁共振(DCE-MRI)在脑胶质瘤(BG)术前诊断与分级的价值.方法:回顾性纳入首发颅内占位性病变经术后病理证实的脑胶质瘤患者42例,术前均行3D-ASL、DCE-MRI扫描,统计DCE扫描下容积转运参数(Ktrans)、血管外细胞外容积分数(Ve)和3D-ASL扫描测量的肿瘤最大血流量值与对侧半球、灰质、白质血流值(rCBF),将患者分为高级别胶质瘤(H组)26例和低级别胶质瘤(L组)16例,对比2组Ktrans、Ve、rCBF参数差异,ROC曲线分析3D-ASL、DCE-MRI在诊断和鉴别BG分级中的效能.结果:Ktrans值和Ve值在BG各级别差异有统计学意义(F=8.264,P＜0.05),其中Ⅰ级和Ⅱ级Ktrans值和Ve值低于Ⅲ级和Ⅳ级(P＜0.05),但在Ⅰ级和Ⅱ级、Ⅲ级和Ⅳ级之间差异无统计学意义(P＞0.05),H组平均Ktrans值和Ve值高于L组(P＜0.05);H组肿瘤各项rCBF均高于L组(P＜0.05);Ktrans值、Ve值、rCBF值均与BG分级呈正相关(r值分别为0.681、0.503、0.635,P＜0.05);Ktrans值、Ve值、rCBF值鉴别BG级别的最佳界值分别为0.058/min、0.362、2.13,其对应的灵敏度、特异度、AUC分别为:96.1%、86.2%、0.926;93.5%、79.2%、0.895;97.2%、97.3%、0.993.结论:3D-ASL在BG分级鉴别中效能优于DEC-MRI.

目的 探讨银杏内酯注射液联合阿替普酶静脉溶栓治疗急性缺血性脑卒中的效果.方法 分析中国人民解放军第二一一医院神经内二科2015年5月～2017年7月收治的106例急性缺血性脑卒中患者临床资料,依据治疗方式进行分组,对照组(阿替普酶治疗)53例和观察组(银杏内酯注射液联合阿替普酶治疗)53例,两组均治疗14 d.观察两组患者治疗前后美国国立卫生研究院卒中量表(NIHSS)评分、改良Rankin量表(mRS)评分、纤维蛋白原(FIB)、凝血酶原时间(PT)、凝血酶时间(TT)、生活质量评分、日常生活能力(Barthel)指数,观察两组患者临床疗效、并发症情况.结果 两组患者治疗前NIHSS评分、改良mRS评分、FIB、PT、TT、生活质量评分和Barthel指数比较,差异均无统计学意义(P>0.05).治疗后,两组患者NIHSS评分、改良mRS评分均低于本组治疗前,生活质量评分和Barthel指数均高于本组治疗前,差异均有统计学意义(P<0.05).治疗后,观察组NIHSS评分、改良mRS评分均低于对照组,生活质量评分和Barthel指数均高于对照组,差异均有统计学意义(P<0.05).对照组治疗后FIB低于治疗前,PT、TT高于治疗前(P<0.05),观察组治疗前后FIB、PT、TT比较,差异均无统计学意义(P>0.05).治疗后,观察组FIB高于对照组,PT、TT低于对照组,差异均有统计学意义(P<0.05).观察组临床治疗总有效率高于对照组、并发症总发生率低于对照组,差异均有统计学意义(P<0.05).结论 银杏内酯注射液联合阿替普酶静脉溶栓治疗急性缺血性脑卒中患者,可以更好地改善患者的神经功能和生活质量,对于凝血功能影响较小,效果明显,并发症少,值得临床推广应用.

目的 探讨常规管腔成像联合磁共振管壁成像技术在头颈部动脉闭塞诊断中的价值.方法 选择2016年5月至2017年5月中国人民解放军211医院收治的60例头颈部动脉闭塞病患者为研究对象,患者均行常规管腔成像和磁共振管壁成像检查,对闭塞血管局部节段的管壁特征以及远端腔内的血栓特征进行分析研究.比较常规管腔成像、常规管腔成像联合磁共振管壁成像技术诊断病因种类的准确率.结果 60例患者共诊断出闭塞血管70段,其中常规管腔成像联合磁共振管壁成像诊断出68段,准确率为97.14％(68/70);常规管腔成像诊断出46段,准确率为65.71％(46/70).常规管腔成像联合磁共振管腔成像技术诊断头颈部动脉闭塞的准确率高于常规管腔成像(x2=10.263,P＜0.05).常规管腔成像联合磁共振管壁成像诊断的68段动脉硬化性血管闭塞中,中动脉硬化性闭塞45段,动脉夹层性闭塞12段,动脉炎性闭塞11段.动脉硬化性闭塞患者闭塞近段的管壁均出现明显的偏心性加厚的现象,动脉夹层性闭塞患者均出现了典型的双腔征或者是新月形管壁内高信号血肿现象,动脉炎性闭塞患者闭塞血管中段出现明显的增厚和强化现象.结论 常规管腔成像联合磁共振管壁成像技术,可充分显示出头颈部动脉闭塞的起始位置的动脉管壁特征,对于诊断动脉硬化性闭塞、动脉炎性闭塞以及动脉夹层性闭塞的病因具有一定的优势.

目的 探讨尿激酶静脉溶栓与超选择性动脉溶栓治疗急性大脑中动脉脑梗死的效果.方法 选择2015年4月～2017年6月解放军第二一一医院神经内二科收治的104例急性大脑中动脉脑梗死患者的临床资料,依据治疗方式不同将其分为尿激酶静脉溶栓组(n=52)和超选择性动脉溶栓组(n=52).观察并比较两组患者溶栓前、溶栓后7d美国国立卫生研究院卒中量表(NIHSS)评分、凝血功能、血管再通率、满意度和并发症的发生率.结果 两组溶栓前NIHSS评分比较,差异无统计学意义(P＞0.05),溶栓后7d两组NIHSS评分均低于溶栓前,超选择性动脉溶栓组溶栓后7 d NIHSS评分低于尿激酶静脉溶栓组,差异均有统计学意义(P＜0.05).两组患者溶栓前、溶栓后7d纤维蛋白原(FIB)、凝血酶原时间(PT)及凝血酶时间(TT)比较,差异无统计学意义(P＞0.05),超选择性动脉溶栓组患者血管再通率、总有效率、患者满意度均高于尿激酶静脉溶栓组,并发症发生率低于尿激酶静脉溶栓组,差异均有统计学意义(P＜0.05).结论 超选择性动脉溶栓治疗可明显改善急性大脑中动脉脑梗死患者神经功能,溶栓效果较好,并发症少,值得临床推广应用.

目的:探讨三维动脉自旋标记成像(3D-ASL)全脑灌注成像技术在脑胶质瘤中的应用价值.方法:前瞻性选取医院收治的18例颅内胶质瘤患者,其中10例高级别胶质瘤、8例低级别胶质瘤患者.测量3D-ASL血流灌注图像肿瘤实质部分最大肿瘤血流量及对侧半球、白质、灰质的脑血流量,分析不同脑肿瘤的磁共振成像(MRI)灌注情况;通过MRI常规扫描序列做出脑肿瘤初步诊断联合3D-ASL灌注成像技术再次诊断后,将两种方法的诊断符合率进行比较.结果:3D-ASL灌注成像技术所得rCBF定量参数在低级别胶质瘤与高级别胶质瘤之间差异有统计学意义(t=5.03,t=4.65,t=4.89;P<0.05).通过常规MRI扫描得出的脑肿瘤定性诊断的符合率为88.9％,联合应用3D-ASL灌注成像技术的诊断符合率为100％,两种方法差异无明显统计学意义.结论:3D-ASL能定量反映肿瘤微循环的灌注情况,与MRI常规序列联合应用,可作为其重要补充,对脑肿瘤的定性诊断以及胶质瘤的术前分级有重要参考价值.

Tumor necrosis factor-a-induced protein 8 (TNFAIP8) presented a elevated expression in endometrial cancer (EC). However, the relationship of TNFAIP8 gene polymorphisms with EC risk remains unclear. This case–control study aimed to investigate the effect of single nucleotide polymorphisms (SNPs) in TNFAIP8 on northern Chinese women with EC. SNP rs11064, rs1045241, and rs1045242 in TNFAIP8 were successfully genotyped in 248 cancer-free controls and 226 ECs by SNaPshot method, respectively. Logistic regression was performed to assess relationship of SNPs with EC risk. The relationships of SNPs with clinicopathological variables were evaluated by Chi-square test or Student’s t-test or Fisher’s text. The minor alleles of rs11064 in TNFAIP8 were strongly associated with EC risk, with adjust odds ratio (OR) of 1.719 (95% CI 1.180–2.506, P = 0.005). The minor allele of rs1045242 in the TNFAIP8 gene was strongly associated with with EC risk (adjust OR: 1.636, 95% CI 1.107–2.417, P = 0.014). rs11064 SNPs correlated with TNFAIP8 protein expression in EC (P = 0.015). For rs1045242, patients with AG + GG presented higher TNFAIP8 protein expression than that with AA (P = 0.020). It also showed that SNP rs11064 was associated with advanced FIGO stage (P = 0.001), deep myometrial invasion (P = 0.047), and lymph node metastasis (P = 0.048) under the codominant model in ECs. SNP rs11064 in TNFAIP8 increased EC risk and significantly related with its protein expression in northern Chinese women.

The role of immune response has gradually become a key factor in the regulation of various complex cancers.Immunotherapy has also become an effective means of cancer cancer intervention. Tumor microenvironment contains different types of immune cells. This helps to regulate the fine balance between innate and adaptive immune systems in anti-tumor signals. In this context, the mechanism of the interaction between tumor cells and immune cells remains to be elucidated. However, it has been proved that many kinds of microRNA play a regulatory role in the development and function of immune cells associated with solid tumors. It enhances or suppresses the immune response through tumor and immune cell mediated immunosuppressive or immunostimulatory factor secretion, which target to tumor associated immune regulatory pathway. Thus playing a key role in all stages of cancer initiation and metastasis, and to find a new therapeutic target in tumor immunotherapy. This article reviews the regulation of microRNA in tumor immune response.

PR domain 14 (PRDM14)has a PR domain and six zinc finger structures,and is one of the representa-tive members of the PRDM family. PRDM family is associated with human tumor formation and plays an important role in cell differentiation and malignant transformation. Each member of PRDM family has a PR-domain region,which has high homology with SET-domain. At home and abroad,it has been found that PRDM family members are abnor-mally expressed during the development of various tumors. In recent years,studies on PRDM14 and tumor gradually have increased,showing its different roles in different tumor types,studies have shown that PRDM14 can also serve as a good drug target malignant tumors. In this paper,the structure and function of PRDM gene were introduced,and the significance of PRDM14 in tumorigenesis and its related research progress was expounded.

Nogo receptor (NgR) play important role of inhibition axonal regeneration after central nervous system (CNS) injury. Rat retinal ganglion cells were transfected with AAV-NgR1siRNA in vivo and vitro to investigate its effect on axonal regrowth. NgR protein expression was assayed by western blot. The sections of cultured RGCs and optic nerve (ON) were stained with GAP-43 antibody. There is little expression of NgR after transfection of AAV-NgR1siRNA compared with control group. There was a significant difference of axonal length between experimental and control group. The present finding indicates that NgR genes play an inhibitive role in the axonal regeneration of RGCs, while transfection of AAV-NgR1siRNA is an effective way to enhance axonal regeneration of ON after injury.

目的:分析新辅助化疗联合间隔肿瘤细胞减灭术(NACT-IDS)和初次肿瘤细胞减灭术(PDS)两种方法在治疗晚期上皮性卵巢癌时对其耐药性和生存时间的影响,并分析铂类耐药性的影响因素.方法:回顾性分析2008年1月至2014年6月就诊我院的354例晚期(FIGOⅢC～Ⅳ期)上皮性卵巢癌患者.根据治疗方法不同分为NACT-IDS组173例和PDS组181例,术后均辅以铂类药物化疗.全部治疗结束后,比较NACT-IDS组及PDS组患者对铂类药物的敏感性,并通过Logistic回归对铂类耐药的单因素、多因素影响进行分析,利用Kplan-Meier方法进行生存分析.结果:至随访结束,PDS组有149例(82.3％)复发,NACT-IDS组有132例(76.3％)复发,两组比较差异无统计学意义(P =0.162).铂类耐药患者PDS组有26例(14.3％),NACT-IDS组有42例(24.3％),两组比较差异有统计学意义(P =0.018).对铂类耐药性作单因素分析发现治疗方法、FIGO分期、总化疗周期与铂耐药性有关(P＜0.05).而多因素分析发现新辅助化疗不是影响铂耐药性的独立危险因素.NACT-IDS组与PDS组中位无病生存期(DFS)(13.5月vs 15.4月,P=0.475)、总生存期(OS)(51.7月vs 46.2月,P=0.147)比较,差异均无统计学意义.结论:NACT-IDS治疗晚期卵巢癌时增加了对铂类药物的耐药性,但对生存时间无明显影响.新辅助化疗不是铂类耐药性的独立危险因素.

Objective:To study the expression and clinical significance of miRNA-34a in cervical exfoliated cells of HPV persistence infection patients.Methods:Poly real(A) tail method SYBR green Ⅰ real-time fluorescent quantitative PCR was performed to examine the expression of miRNA-34a on cervical exfoliated cells in 60 patients,and analyzed its relations with HPV persistent infection.Results:The expression level of miRNA-34a of HPV persistent infection group was down-regulated in cervical exfoliated cells as compared with the HPV infection outcome group(P ＜ 0.05).Conclusion:miRNA-34a has certain relation with HPV persistent infection.And is expected to become a new indicator for HPV persistent infection in early cervical cancer screening,diagnosis and clinical diagnosis and treatment.

Persistent infection with human papillomavirus (HPV), particularly type 16, is causally associated with cervical cancer and its precursors. The role of miRNAs in HPV16 persistence currently remains unclear. Preliminary analysis of miRNA profile demonstrated that HPV16 infection caused a striking downregulation of miR-34a. Through bioinformatics analysis and dual-luciferase assay with site-directed mutagenesis strategy, NLRC5, a negative regulator of NF-κB signaling, was identified to be a novel interactor of miR-34a. Transfection of miR-34a mimic strikingly downregulated NLRC5 in the HPV16-positive cervical cells, which might result in the nuclear accumulation of NF-κB p65. However, transfection of miR-34a inhibitor exhibited an opposite effect. The antagonistic expressions of NLRC5 and miR-34a were also observed in keratinocytes harboring HPV16 genome as well as in human cervical samples with persistent infection of HPV16. Our data uncover a previously unknown connection among HPV16 persistence, miR-34a and its interactor NLRC5.

分化抑制因子-1(inhibitor of differentiation-1,Id1),属于螺旋-环-螺旋(helix-loop-helix,HLH)蛋白,研究已经证实,Id1蛋白的异常高表达与人类恶性肿瘤的增殖、侵袭和转移关系密切,是影响肿瘤患者预后的重要影响因素,同时为抗肿瘤药物提供了新的作用靶点.本文针对Id1基因与妇科肿瘤的相关性研究做一综述.

目的:通过构建人卵巢癌SKOV3 Id1特异性siRNA慢病毒载体及其裸鼠移植瘤模型,观察沉默分化抑制因子(inhibitcr of differentiation-1,Id1)表达对卵巢癌SKOV3生长的影响.方法:根据Id1基因序列设计并合成的siRNA片段转染SKOV3细胞,将转染细胞分为四组:cell+ lv-shRNA-Id1(实验组)、cell+ lv-shR-NA-NC(质粒对照组)、cell+ lv-control(空病毒对照组)、cell(空白对照组),筛选出稳定转染的SKOV3细胞克隆,48小时后检测细胞生长活性.将40只裸鼠随机分为四组,分别接种上述转染成功的细胞,30天后称取瘤体重量,计算抑瘤率;然后通过Western blotting法检测移植瘤中Id1蛋白的表达.结果:在体外试验中,实验组的细胞生长活性显著低于空白对照组,具有统计学意义(P＜0.05),三对照组间无明显差异(P＞0.05);体内实验显示,实验组平均瘤重均低于其他三对照组,差异具有统计学意义(P＜0.05),三对照组间瘤重差异无统计学意义(P＞0.05);实验组、质粒对照组、空病毒组抑瘤率分别为45.98％、4.84％、4.50％;Westernblotting结果显示,实验组中Id1表达较对照组明显下调(P＜0.05).结论:沉默Id1基因能抑制卵巢癌细胞的生长,有望成为卵巢癌治疗新靶点.

[目的]探讨过氧化物酶体增殖物激活受体γ(PPAR-γ)配体罗格列酮(RGZ)对人卵巢浆液性乳头状囊腺癌SKOV3细胞的抑制作用.[方法]建立卵巢癌SKOV3细胞裸鼠移植瘤模型,观察不同剂量RGZ对移植瘤体积的影响;HE染色观察各组移植瘤组织细胞形态学变化;免疫细胞化学法观察移植瘤中Bcl-2、Cytochrome C蛋白表达水平.[结果]RGZ组(4mg/kg,12mg/kg,60mg/kg,120mg/kg)裸鼠移植瘤体积分别为0.190±0.023mm3、0.065±0.003mm3、0.195 ±0.004mm3和0.182±0.023mm3,较对照组(0.300±0.016mm3)均明显缩小,差异有统计学意义(P＜0.05).抑瘤率分别为36.7％、78.3％、35.0％和40.0％.HE染色观察RGZ作用SKOV3细胞后有变性细胞、单细胞死亡及大面积的凝固性坏死.Bcl-2在RGZ组(60mg/kg,120mg/kg)中的表达较对照组显著下调,Cytochrome C在RGZ组(60mg/kg,120mg/kg)中的表达较对照组显著上调,差异均有统计学意义(P＜0.05).[结论] RGZ对卵巢癌裸鼠移植瘤有明显抑制作用,可促进凋亡.提示PPAR-γ可能成为卵巢癌治疗的新分子靶点.

[目的]探讨体外DNA结合抑制因子(inhibitor of DNA bindingor differentiation,Id)的表达对卵巢上皮癌SKOV3对顺铂化疗敏感性的影响.[方法]构建靶向Id1基因siRNA慢病毒载体并转染SKOV3细胞,筛选出稳定转染的SKOV3细胞克隆.实验分为4组:实验组(Cell+Lv-shRNA-Id1)、阴性对照组(Cell +Lv-shRNA-NC)、病毒对照组(Cell +Lv-control)与空白对照组(Cellgroup).CCK8检测各组SKOV3细胞不同时间增殖活性以及暴露于不同浓度顺铂(0.1、0.2、0.5、1、2、5、10μg/ml) 48h后顺铂对各组SKOV3细胞的半数抑制浓度,分析沉默Id1基因后卵巢癌细胞对顺铂化疗敏感性的影响.[结果]三组对照组细胞增殖活性随着时间增加显著,而实验组则增加缓慢.在48h和72h时,实验组细胞增殖活性值分别为0.449±0.072μg/ml、0.885±0.232μg/ml,与三组对照组比较差异均有统计学意义(P均＜0.05);实验组顺铂对SKOV3细胞的半数抑制浓度为1.5±0.71 μg/ml,明显低于对照组(P＜0.01).[结论]抑制Id1基因表达可以增加顺铂对卵巢癌细胞的生长抑制作用,为临床进一步提高卵巢癌的疗效提供了研究基础.