宫颈癌患者高危型人乳头瘤病毒感染 与免疫功能和癌基因表达的相关性

摘要：目的 探讨宫颈癌患者高危型人乳头瘤病毒（HPV）感染的检测及其与免疫功能和癌基因表达的相关性。方法 选择2017年9月至2018年12月在我院接受手术治疗的118例原发性宫颈癌患者为宫颈癌组,根据高危型HPV感染情况进一步分为高危型HPV组（89例）和非高危型HPV组（29例）。选择同期在我院行子宫全切术的57例子宫肌瘤患者为子宫肌瘤组。对比各组患者外周血T淋巴细胞亚群分布情况,病灶组织中原癌基因（E6/E7、c-Met、SALL4、PGRN）和抑癌基因（p53、pRb、PTEN、LKB1）表达量的差异。结果 宫颈癌组患者病灶组织中高危型HPV感染率显著高于子宫肌瘤组（75.42% vs 22.81%,2=43.764,P＜0.05）。高危型HPV组患者外周血中CD4+ T淋巴细胞比例、CD4+T/CD8+ T比值低于非高危型HPV组,CD8+ T淋巴细胞比例高于非高危型HPV组（均P＜0.05）。高危型HPV组患者病灶组织中E6/E7、c-Met、SALL4、PGRN mRNA的表达量高于非高危型HPV组,而p53、pRb、PTEN、LKB1 mRNA的表达量低于非高危型HPV组（均P＜0.05）。结论 高危型HPV感染可导致宫颈癌患者细胞免疫功能下降及肿瘤恶性程度增加,可能是导致病情恶化、治疗效果不佳的危险因素之一。     

前列腺癌根治术治疗不同分级前列腺癌患者的临床研究

目的:探究前列腺癌根治术在不同危险度前列腺癌患者中治疗的临床效果,为临床前列腺癌患者的治疗提供依据。方法:选择2008年1月~2015年12月期间我院94例前列腺癌患者为研究对象,根据D'Amico评分将其分为高危、中危及低危三组,收集患者基线资料、术后随访资料,并比较三组手术并发症;采用Kaplan-Meier分析法计算三组患者生存率,并采用Log-rank检验比较不同危险组的生存率。结果:高危组患者进行开放性手术人数多于中危组和低危组,且中危组多于低危组,差异具有统计学意义(P0.05);高危组患者术前Gleason评分和PAS水平高于中危组和低危组,且中危组高于低危组,差异具有统计学意义(P0.05);术后5年高危组患者完全控尿率显著低于中危组和低危组(P0.05);三组患者间5年无生化复发率比较无统计学意义(P0.05)。结论:前列腺癌根治术治疗高危前列腺患者较中、低危患者疗效较差,但仍可达到较好的疗效,可在临床推广使用。     

miR-9靶向调控PTEN在HPV16+及HPV18+宫颈癌细胞凋亡中的作用

研究微小RNA-9(microRNA-9,miR-9)靶向调控人第10号染色体缺失的磷酸酶及张力蛋白同源蛋白（Recombinantphosphataseandtensinhomolog,PTEN）在人乳头瘤病毒（Humanpapillomavirus,HPV）16+及HPV18+宫颈癌细胞凋亡中的作用。收集宫颈癌组织及正常宫颈组织，培养HPV16+SiHa细胞、HPV18+HeLa细胞、HPV阴性C33A细胞及正常宫颈上皮H8细胞，检测miR-9、PTEN的表达水平。SiHa细胞和HeLa细胞进行分组转染后检测增殖抑制率、凋亡率、PTEN及miR-9表达水平。结果显示高危型HPV阳性及阴性的宫颈癌组织中miR-9的表达水平高于正常宫颈组织、PTEN的表达水平低于正常宫颈组织（P<0.05）且高危型HPV阳性宫颈癌组织中miR-9、PTEN表达的变化更显著；SiHa细胞、HeLa细胞中miR-9的表达水平高于C33A细胞和H8细胞、PTEN的表达水平低于C33A细胞和H8细胞（P<0.05）；敲低miR-9表达后，SiHa细胞、HeLa细胞的PTEN表达水平、增殖抑制率、凋...     

宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性

目的探讨宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。方法选择2018年1月至2019年1月间在我院确诊为原发性宫颈癌的患者50例作为宫颈癌组,在我院诊断为宫颈糜烂的患者78例作为宫颈糜烂组,同期在我院进行体检的健康女性100例作为正常对照组。对比3组研究对象的阴道微生态失调率、高危型HPV感染率以及宫颈癌组、宫颈糜烂组患者宫颈病灶组织中宫颈癌相关增殖基因(Prdx4、Nek2、Fhit、BLCAP)mRNA表达量的差异。采用Pearson检验分析宫颈癌患者阴道微生态失调率与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。结果宫颈癌组、宫颈糜烂组患者的阴道微生态失调率、高危型HPV感染率高于正常对照组,其中宫颈癌组患者这两项指标水平高于宫颈糜烂组(均P0.05)。宫颈癌组患者宫颈病灶组织中Prdx4、Nek2 mRNA表达量高于宫颈糜烂组,Fhit、BLCAP mRNA表达量低于宫颈糜烂组(均P0.05)。相关性分析发现,宫颈癌患者阴道微生态失调率与高危型HPV感染率呈正相关,与癌基因(Prdx4、Nek2)mRNA表达量呈正相关,与抑癌基因(Fhit、BLCAP)mRNA表达量呈负相关(均P0.05)。结论宫颈癌患者阴道微生态失调率较高,可能与高危型HPV感染及癌细胞增殖旺盛密切相关。     

APOBEC3G在宫颈癌发生发展中的表达及意义

目的探讨载脂蛋白BmRNA编辑酶催化多肽样蛋白3G(apolipoprotain B mRNA-editing enzyme catalytic polypeptide 3 protein,APOBEC3G)在维吾尔族女性患者宫颈癌(uterine cervical carcinoma,UCC)发生发展的各个阶段中的表达及意义。方法选择2015年1月—2017年12月在新疆维吾尔自治区人民医院妇科门诊和病区进行宫颈液基薄层细胞学检查和HC-Ⅱ高危型HPV检测联合筛查,并同时进行阴道镜活检和组织病理学诊断的维吾尔族女性患者190例,其中高危型人乳头瘤病毒(human papilloma virus,HPV)阴性慢性宫颈炎41例、高危型HPV阳性慢性宫颈炎40例、UCC前病变(宫颈上皮内瘤变I~III级)41例、UCC 68例。提取宫颈组织中的RNA和蛋白质,并采用Real-Time PCR检测APOBEC3G mRNA转录水平,采用Western blot检测APOBEC3G蛋白表达水平。结果APOBEC3GmRNA在HPV阴性宫颈炎、HPV阳性宫颈炎、UCC前病变、UCC各组表达水平差异均有统计学意义(F=11.00,P=0.000 1);组间两两比较,UCC组表达量是HPV阴性宫颈炎组的1.70倍,HPV阳性宫颈炎组的1.33倍,差异均有统计学意义(P0.05);UCC前病变组表达量是HPV阴性宫颈炎组的3.00倍,HPV阳性性宫颈炎组的1.65倍,差异均有统计学意义(P0.05);APOBEC3G在HPV阴性宫颈炎、HPV阳性宫颈炎、UCC前病变、UCC各组蛋白表达水平差异均有统计学意义(F=213.15,P=0.000 1);组间两两比较,HPV阴性宫颈炎、HPV阳性宫颈炎、UCC前病变、UCC组间差异均有统计学意义(P0.05)。结论 APOBEC3G在维吾尔族女性患者UCC发生发展各个阶段均有不同程度的表达,提示APOBEC3G在其UCC发展进程中发挥一定作用。     

P21蛋白在不同级别宫颈组织中的表达及其与高危HPV感染相关性的研究

目的通过检测宫颈组织中p21waf基因表达与高危HPV感染的情况,研究P21蛋白与高危HPV感染在宫颈组织恶性转化过程中的作用及其相互关系。方法应用免疫组化检测P21蛋白及基因杂交捕获Ⅱ代技术(HC-Ⅱ)检测高危HPV在正常宫颈组、宫颈炎、宫颈上皮内瘤样病变(CIN)及宫颈癌组这4组中的表达情况。结果在正常宫颈组、宫颈炎、宫颈上皮内瘤样病变(CIN)、及宫颈癌组中P21的阳性表达率分别为11.8%、15.4%、39.1%和57.7%;高危HPV的阳性表达率分别为20%、23.5%、65.2%和88.5%,这两项指标在CIN、宫颈癌组有明显的升高趋势且与正常宫颈和宫颈炎两组相比差异具有显著性统计学意义(P<0.05);各级别宫颈组织中高危HPV阳性组的P21蛋白阳性率明显高于阴性组的P21蛋白阳性率。差异在统计学上具有显著性意义(P<0.05)。结论P21蛋白的表达和高危HPV感染与宫颈病变的恶化均有高度的相关性,其检出率和阳性表达率随着宫颈病变恶性程度的增加而升高。在CIN向宫颈癌恶性转化过程中,P21与高危HPV共同促进肿瘤的发生。     

宫颈癌细胞系中HPV病毒感染与P16和E-cadherin表达的相关性分析

目的探讨宫颈癌细胞系中HPV感染状态与P16和E-cadherin表达之间的相关性。方法应用免疫细胞化学染色、免疫荧光、Western-blot以及RT-PCR的方法检测HeLa、SiHa和C33A三个细胞系中P16和E-cadherin的表达情况。结果HPV阳性的两个细胞系HeLa和SiHa中P16的表达呈强阳性而E-cadherin的表达呈弱阳性,HPV阴性的细胞系C33A中P16的表达为弱阳性而E-cadherin呈强阳性表达。结论宫颈癌细胞系中HPV的感染与P16的表达呈正相关而与E-cadherin的表达呈负相关。     

宫颈癌中高危型HPV感染与miR-362-3p、Nemo样激酶表达的相关性研究

高危型人乳头瘤病毒(High-risk human papillomavirus,HPV)感染是宫颈癌发病的重要因素,但高危型HPV感染引起宫颈上皮恶变的机制尚不清楚.微小RNA (micro RNA,miR)-362-3p是具有抑癌活性的miR,在宫颈癌中表达降低;Nemo样激酶(Nemo-like kinase,NLK)是生物信息学预测得到的miR-362-3p靶基因,在宫颈癌中表达增加.但宫颈癌发病过程中高危型HPV感染与miR-362-3p、NLK的关系尚不清楚.本研究检测了miR-362-3p、NLK在宫颈癌组织及宫颈癌细胞株中的变化,并通过转染miR-362-3p模拟物、NLK表达质粒的方式验证了miR-362-3p靶向NLK调节高危型HPV阳性宫颈癌细胞增殖的作用.结果 显示:与癌旁组织比较,宫颈癌组织中miR-362-3p的表达明显降低、NLK的表达明显增加且与高危型HPV阴性的宫颈癌组织比较,高危型HPV阳性的宫颈癌组织中miR-362-3p的表达明显降低、NLK的表达明显增加;与正常宫颈上皮细胞比较,HPV16感染的SiHa细胞、HPV18感染的HeLa细胞及HPV阴性的C33A细胞中miR-362-3p的表达明显降低、NLK的表达明显增加且SiHa细胞中miR-362-3p表达降低、NLK表达增加最明显;在SiHa细胞中,过表达miR-362-3p能够降低细胞活力及NLK的表达,同时也使NLK双荧光素酶报告基因的荧光活力降低;过表达NLK能够逆转miR-362-3p降低细胞活力及NLK表达的作用.以上结果表明宫颈癌中高危型HPV感染与miR-362-3p低表达、NLK高表达有关,过表达miR-362-3p通过靶向抑制NLK降低高危型HPV感染宫颈癌细胞的增殖活力.本研究的创新点在于初步探索了高危型HPV感染促进宫颈癌细胞增殖的作用及机制,在高危型HPV感染的宫颈癌中miR-362-3p表达减少并靶向引起NLK表达增加,进而促进了宫颈癌细胞的增殖,这为将来深入认识高危型HPV引起宫颈癌发病的机制提供依据.     

血清肿瘤标志物在肺癌辅助诊断中的应用

目的探讨血清肿瘤标志物在肺癌诊断中的临床价值。方法收集40例健康人、45例肺部良性疾病患者和90例肺癌患者,采用电化学发光分析检测患者血清中肿瘤标志物细胞角蛋白19(CYFRA21-1)、鳞状细胞癌相关抗原(SCC)和癌胚抗原(CEA),以及胃泌素释放肽前体(pro-GRP)和神经元特异性烯醇化酶(NSE)的含量。结果健康人组和肺部良性疾病患者组血清NSE、pro-GRP、CYFRA21-1、SCC和CEA水平较肺癌患者组水平低,差异有统计学意义(P0.01)。NSE和pro-GRP在小细胞肺癌患者中的水平均明显高于其他类型的肺癌患者(P0.01),CYFRA21-1和SCC在鳞癌患者中的含量比其他类型肺癌患者高(P0.01)。联合检测此5种血清肿瘤标志物敏感性高于单独的肿瘤标志物(P0.01)。结论联合检测NSE、pro-GRP、CYFRA21-1、SCC和CEA可以提高肺癌诊断的灵敏度。     

HPV16 E6对HeLa细胞中Rap1GAP蛋白水平下调的研究

目的探讨HPV16E6对细胞中Rap1GAP蛋白水平的影响,为阐明宫颈癌发生发展的分子机制提供实验依据。本研究组前期实验显示,宫颈癌石蜡切片组织中Rap1GAP蛋白水平下降,且与高危型HPV16/18感染相关。本文将进一步探讨HPV16 E6是否导致Rap1GAP蛋白下调的原因。方法通过将HPV16 E6基因插入pGEX-KG的BamHI和Hind Ⅲ酶切位点构建GST标记的HPVE6质粒,采用脂质体转染法将其转染入HeLa细胞中,Westernblot方法观察GST-tagged-HPV16 E6在HeLa细胞中的表达,并观察其对HeLa细胞中内源性Rap1GAP蛋白水平的影响。结果测序表明成功构建GST标记的HPV16E6质粒;Western blot检测表明HPV16E6在HeLa细胞中成功表达;并且发现HeLa细胞中过表达HPV16 E6后,Rap1GAP蛋白相对含量(0.602±0.205)明显低于未转染组(1.130±0.163),差异具有统计学意义(P0.05)。结论HPV16 E6下调Rap1GAP蛋白水平。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

Scurvy in capybaras bred in captivity in Argentine

In order to determine if the absence of vitamin C in the diet of capybaras (Hydrochoerus hydrochaeris) causes scurvy, a group of seven young individuals were fed food pellets without ascorbic acid, while another group of eight individuals received the same food with 1 g of ascorbic acid per animal per day. Animals in the first group developed signs of scurvy-like gingivitis, breaking of the incisors and death of one animal. Clinical signs appeared between 25 and 104 days from the beginning of the trial in all individuals. Growth rates of individuals deprived of vitamin C was considerably less than those observed in the control group. Deficiency of ascorbic acid had a severe effect on reproduction of another population of captive capybaras. We found that the decrease in ascorbic acid content in the diet affected pregnancy, especially during the first stages. The results obtained suggest that it is necessary to supply a suitable quantity of vitamin C in the diet of this species in captivity.     

Changes in lactate dehydrogenase activity in chicken tissues in hyperthyreosis in ontogenesis

The lactate dehydrogenase activity in reactions of lactate oxidation and synthesis was studied in subfractions of the chicken brain, heart and liver at the embryonal, early postembryonal and adult stages of development after thyroxine administration. It has been shown that during embryogenesis thyroxine predominantly enhanced the rate of lactate oxidation in the mitochondrial tissues. A marked increase in the lactate synthesis was found in cytoplasm of the adult chicken tissues. Specificity of enzyme activity alterations was detected in the chicken brain during ontogenesis after thyroxine administration.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.