某些环境促癌因素的实验研究——Ⅰ乌桕与了哥王对HSV_2诱癌的促进作用

为了研究可疑促癌剂乌桕及了哥王提取物的作用,我们采用我室HSV_2诱发小白鼠宫颈癌的动物模型,进行体内促癌实验。这篇论文报道了实验结果。小白鼠分成6组,每组癌发率如下:HSV_2+0.9%;NaCl:13.0％;HSV_2+乌桕:40.0％;HSV_2+了哥王:36.4％;HSV_2+巴豆油:55.0％;乌桕+0.9％NaCl:7.7％;了哥王+0.9％NaCl:0。经过x~2检验指出差别在统计学上具显著性。因此我们提出:乌桕和了哥王提取物对小鼠宫颈癌有促进作用。与巴豆油提取物比较,其中巴豆油作用较强,乌桕作用次之,了哥王作用较弱。     

黄芫花及桐油提取物对Ⅱ型单纯疱疹病毒诱癌的促进作用

本文报道,瑞香科植物黄芫花Wikstroemia chamaedaphne(WC)提取物,及大戟科桐油提取物12-O-hexadecanoyl-16-hydroxyphorboI-13-acetate(HHPA),能促使Ⅱ型单纯疱疹病毒(HSV-2)333株诱发小鼠宫颈癌,其癌发率分别为21.1％和26.3％,而对照组的癌发率仅为7.4％,统计学处理(累积法及权重法计算后,用方差分析),其差异均有显著性,证明WC和HHPA是促癌物。     

长链非编码RNA与宫颈癌关系的研究进展现,维

LncRNA是一类长度大于200个碱基的非编码RNA分子,广泛参与真核细胞内基因的表达与调控,在维持机体正常的生理功能中发挥重要作用。lncRNA表达异常与包括肿瘤在内的多种疾病的发生与发展密切相关。在宫颈癌细胞中,部分lncRNA发挥肿瘤抑制因子的作用,在癌组织中表达沉默,而另一些lncRNA则起促癌因子的作用,在癌组织中表达显著性上调。这些lncRNA的异常表达通过影响与肿瘤发生发展相关的信号分子活性以及细胞周期、细胞凋亡和上皮细胞间质转化等过程而促进宫颈癌细胞的恶性行为。LncRNA异常表达与宫颈癌的恶性表型、FIGO分级、侵袭转移和不良预后密切相关,是宫颈癌诊断与预后的重要指标和潜在的治疗靶点。     

瘦素对大鼠主动脉血管平滑肌细胞增殖和移行的影响

目的本实验旨在研究瘦素(Leptin)对大鼠主动脉血管平滑肌细胞(VSMC)增殖和移行的影响。方法采用原代细胞培养方法建立VSMC细胞模型;以四甲基偶氮唑盐(MTT)比色法、免疫细胞化学染色等方法观察瘦素对VSMC增殖的影响;采用细胞移行实验观察瘦素对VSMC移行的影响;采用MAPK的抑制剂SB203580观察瘦素对VSMC增殖的抑制作用以探讨MAPK在瘦素促VSMC增殖中的信号转导机理。结果瘦素能显著促进VSMC增殖和移行,而MAPK的抑制剂SB203580可显著抑制瘦素对VSMC的增殖效应。结论瘦素具有明显的促VSMC增殖和移行的作用,其作用机制与激活MAPK信号途径有关。     

黄芫花提取物对V_(79)细胞和WB肝细胞的生物学效应 1.细胞连接通讯与增殖活性的相关性变化

用SLDT技术、细胞生长曲线、~3H-TdR掺入、细胞分裂指数和流式细胞光度术等方法在V_(79)细胞和WB肝细胞中观察到中药促癌物黄芫花提取物刺激细胞增殖的作用与对细胞GJIC抑制的作用有平行相关性。黄芫花提取物抑制V_(79)细胞的GJIC,同时刺激DNA合成,使细胞周期M期时相百分比升高,与TPA的效应相似,对黄芫花提取物促癌作用机理、GJIC抑制与细胞增殖周期的相关性进行分析讨论。     

CLDN6对宫颈癌细胞凋亡、凋亡相关蛋白及Akt信号通路活性的影响

目的研究紧密连接蛋白6(claudin 6,CLDN6)对宫颈癌细胞凋亡的影响。方法收集46例宫颈癌患者宫颈癌组织及对应的癌旁组织,应用免疫组织化学、Western blot和RT-PCR检测宫颈组织中CLDN6的表达水平;应用Annexin V/PI双染法流式细胞术检测过表达CLDN6对人宫颈癌细胞系Si Ha细胞凋亡的影响;Western blot检测过表达CLDN6对Si Ha细胞Bcl-xl、Bax、Bcl-2、Akt和p-Akt等蛋白水平的影响。结果宫颈癌组织中CLDN6 m RNA水平及蛋白水平均明显低于癌旁组织;过表达CLDN6使Si Ha细胞凋亡率和促凋亡蛋白Bax水平明显增加,抑凋亡蛋白Bcl-2和Bcl-xl水平和Akt磷酸化水平明显降低。结论 CLDN6可能通过抑制Akt信号通路活性促进宫颈癌细胞凋亡,宫颈癌的发生可能与CLDN6表达减少有关。     

长链非编码RNA Dleu2 对宫颈癌细胞增殖、迁移及侵袭的影响

目的:研究长链非编码RNA(long non-coding RNA,lnc RNA)Dleu2对宫颈癌细胞增殖、迁移和侵袭能力的影响。方法:利用高通量Lnc RNA芯片技术检测10例宫颈癌组织及对应的癌旁组织,筛选得到一批表达水平具有显著差异的Lnc RNA,进一步针对可能具有生物学功能的Lnc RNA-Dleu2,利用q-PCR验证其在癌组织样本中的相对低表达。再通过在细胞内过表达Lnc RNA-Dleu2研究其对宫颈癌Hela和Caski细胞系增殖、迁移和侵袭的影响。结果:q-PCR结果验证了Lnc RNA芯片筛选的结果,即相较于癌旁组织和正常宫颈上皮细胞系,Lnc RNA-Dleu2在宫颈癌组织和细胞中均低表达。CCK8和克隆形成实验结果显示,过表达Lnc RNA-Dleu2能显著抑制Hela和Caski细胞增殖能力(P0.01);细胞划痕实验结果显示,过表达Lnc RNA-Dleu2能显著抑制Hela和Caski细胞增殖和迁移能力;Matrigel细胞侵袭实验结果显示,过表达Lnc RNA-Dleu2能显著抑制Hela和Caski细胞的侵袭能力(P0.01)。结论:Lnc RNA-Dleu2在宫颈癌中相对低表达,提高Dleu2表达水平能够抑制宫颈癌细胞系Hela和Caski的增殖、迁移和侵袭能力。     

双香豆素在HBx介导的肝癌细胞生长增殖中的作用及其机制研究

HCC是世界范围内最常见的恶性肿瘤之一,其中50％以上与乙型肝炎病毒(HBV)感染相关,且乙肝相关性肝癌患者术后复发率明显高于未合并乙肝患者.而病毒蛋白HBx是促进乙肝相关性肝癌进展的主要因素.因此,寻找能够有效抑制HBx促癌作用的小分子化合物,减缓乙肝相关性肝癌的疾病进展,对于延长患者生存期具有重要意义.该研究初步探...     

HPV-16癌蛋白促宫颈癌血管生成及其机制

多种肿瘤组织中有人乳头瘤病毒(human papillomavirus,HPV)相关癌蛋白的检出,尤其是宫颈癌活检组织中高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)癌蛋白的检出率高达95%。近年的研究发现,HR-HPV癌蛋白(特别是HPV-16E6和E7)与宫颈癌血管生成有关,其机制主要是:(1)HR-HPV癌蛋白可以增强一些血管生成促进因子,如血管内皮生长因子(vascular endothelial growth factor,VEGF)、缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)等的表达从而促进血管生成;(2)HR-HPV癌蛋白也可以降低一些血管生成抑制因子,如血小板应答蛋白(thrombospondin,TSP)、乳腺丝酶抑制蛋白(mammary serine protease inhibitor,maspin)等的表达进而使血管生成免受抑制。本文对HPV-16癌蛋白及其与宫颈癌血管生成的关系作一综述。     

宫颈癌中相关基因启动子高甲基化研究进展

本文主要从分子水平介绍了在宫颈癌发生发展过程中抑癌基因的甲基化的作用.以往认为人乳头状病毒高危型的的感染是导致宫颈癌发生的主要因素.随着科技的发展,宫颈癌组织中抑癌基因的甲基化越来越备受关注.既往认为基因内突变和染色体物质缺失是肿瘤抑制基因失活的主要原因.但是,启动子CPG岛异常甲基化导致的基因失活在肿瘤发生发展过程中起着非常重要的作用现已确切证明,DNA甲基化是肿瘤抑制基因失活的第三种机制,而且在某些情况下是抑癌基因失活的惟一机制.对宫颈癌组织中的对相关抑癌基因甲基化的筛选并作为标记应用在检测宫颈癌中,这在防止宫颈癌的发生起到重大作用,还可有望作为宫颈癌治疗疗效检测的一项手段.     

Efficiency of Various Growth Media in Recovering Oral Bacterial Flora from Human Dental Plaque

MM10 sucrose blood agar (MM10 SB agar), N(2)C agar, Schaedler agar (SH agar), and mitis salivarius agar (MS agar) were tested for their ability to recover human dental plaque flora by a continuous anaerobic procedure and by a conventional anaerobic method. MM10 SB agar yielded higher recovery of bacteria from plaque samples as determined by the enumeration of colony-forming units (CFU). The CFU on N(2)C agar, SH agar, and MS agar were lower than MM10 SB agar when the continuous anaerobic procedure was used. The superior performance of MM10 SB agar was much more apparent when used for the cultivation of dental plaque by the conventional anaerobic method. Under these conditions the counts were consistently higher on MM10 SB agar as compared to the other media tested. However, the differential counts of Streptococcus sanguis and S. mutans from carious plaque samples were in general comparable on all culture media. Deletion of blood from MM10 SB agar did not lower counts. The elimination of dithiothreitol from this medium resulted in a significantly lower recovery of bacteria from the plaque samples when cultured by the conventional anaerobic method. The storage of MM10 SB agar for varying periods of time aerobic conditions did not seem to affect its performance. These findings suggest that MM10 SB agar is an ideal culture medium for the isolation, nonselective enumeration, and differential counts of bacteria present in normal and disease-associated plaques.     

宫颈癌中p53表达和病毒感染的研究

应用PCR对121例宫颈脱落细胞和组织分别检测HPV-16／18和HSV-2DNA。同时应用免疫组化S-P法检测76例宫颈组织中P53过度表达。结果发现,宫颈癌组织中HPV-16／18和HSV-2阳性率分别为61．3％和32．3％,慢性宫颈炎组HPV-16／18和HSV-2阳性率分别为22．5％和20．0％,与正常宫颈组比较均有显著性差异。宫颈癌中HPV-16／18和HSV-2混合感染率为16．1％。p53过度表达率在慢性宫颈炎、宫颈上皮内瘤变(CIN)、宫颈癌组织中呈梯度递增。另外,宫颈癌组织中P53过度表达与HPV-16／18、HSV-2的感染无相关性。提示：宫颈癌的发生与HPV-16／18关系密切,HSV-2可能与HPV-16／18协同作用导致宫颈癌的发生。宫颈组织中p53过度表达率与宫颈癌的进程有关,这在宫颈癌的防治方面有一定的临床意义。     

老年人胃内菌群研究

目的研究人体胃内菌群,探讨老年与非老年人胃内菌群的差异。方法选择67例无严重胃肠道疾病的患者为研究对象。其中男性50例、女性17例。年龄≥60岁53例,〈60岁14例。胃镜下取胃组织及胃液,测胃液pH,并做胃组织需氧、厌氧细菌培养及真菌培养,计数胃组织细菌培养数量。16SrRNA法鉴定胃组织细菌种类。真菌的鉴定按微生物科常规菌种方法鉴定。结果老年人中胃内需氧细菌培养阳性为23例（48.93%）,12例（25.53%）胃内需氧菌培养〉1×10^5CFU/g;厌氧细菌培养阳性为22例（46.81%）,12例（25.53%）胃内厌氧菌培养〉1×10^5CFU/g。非老年人中需氧细菌培养阳性为4例（28.57%）,1例（7.14%）胃内需氧菌培养〉1×10^5CFU/g;厌氧细菌培养阳性为4例（28.57%）,1例（7.14%）胃内厌氧菌培养〉1×10^5CFU/g。但老年人与非老年人比较,细菌培养阳性率及细菌培养〉1×10^5CFU/g的比率差异无显著性。仅1例老年人胃组织分离出真菌,为白色念珠菌。胃内共分离出细菌69株,其中革兰阳性球菌31株（44.93%）,革兰阳性杆菌12株（17.39%）,革兰阴性球菌11株（15.94%）,革兰阴性杆菌15株（21.74%）。需氧菌13株（18.84%）,需氧兼性厌氧菌54株（78.26%）,专性厌氧菌2株（2.90%）。老年人胃内常见的细菌是：链球菌、大肠埃希菌、奈瑟菌;非老年人胃内常见的细菌是：链球菌和大肠埃希菌。多为口咽部和胃肠道常见菌群,部分为条件致病菌。结论约46%～48%的老年人胃内细菌培养阳性,约25%的老年人有胃内细菌过度生长（〉1×10^5CFU/g）。约28%的非老年人胃内细菌培养阳性,约7%的非老年人有胃内细菌过度生长。老年人胃内菌群分布与非老年人相似,为口咽部和胃肠道常见菌群,部分为条件致病菌。     

Anaerobic seminal fluid micro-flora in chronic prostatitis/chronic pelvic pain syndrome patients

We investigated the seminal micro-flora of 116 men. Eighty-four men had chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), and 34 of them were also leukocytospermic. Thirty-two asymptomatic men formed the control group. Micro-organisms were found in all of the 116 seminal fluid specimens. More than 20 different micro-organisms were found in both groups. Neisseria gonorrhoeae and Chlamydia trachomatis were not found. A high frequency of anaerobic bacteria was found in all groups (68-79%), and in most of the specimens, anaerobic micro-organisms were equal to or outnumbered the aerobic strains. We found 1-8 different micro-organisms in each semen sample, the total count of micro-organisms ranged from 10(2) to 10(7)/mL of semen. Both parameters were significantly higher in leukocytospermic CP/CPPS (NIH IIIA category) patients (median=5 different micro-organisms; total median count 5 x 10(4)) than in the control group (median=3 different micro-organisms; total median count 10(3)). In the CP/CPPS patients, the prevalence and/or count of some opportunistic bacteria was higher than in the control group. To show that the micro-organisms do not originate from the urethra, first voided urine was also investigated in 17 prostatitis patients and 15 controls. One patient had significantly fewer micro-organisms (median 1 vs. 4) and a lower total count of micro-organisms (median 10(2) vs. 10(4)/mL) in the first-catch urine than in the seminal fluid. We found only one third of the micro-organisms to be similar in urine and semen while anaerobic bacteria and some aerobic opportunists were infrequent in urine. Semen is a suitable specimen for the diagnosis of prostatitis.     

慢性前列腺炎患者前列腺液的微生物研究

目的探讨慢性前列腺炎患者前列腺内的微生物种类。方法慢性前列腺炎81例,其中Ⅱ型前列腺炎（慢性细菌性前列腺炎）11例,Ⅲ型前列腺炎（慢性非细菌性前列腺炎）70例。前列腺液取材行细菌培养、解脲脲原体（UU）和人型支原体（MH）的培养、沙眼衣原体（CT）检测,并行淋病奈瑟菌（NG）、结核分枝杆菌（TB）、UU、CT、HPV、HSV-2、CMV的PCR检测,以及细菌16SDNA检测。结果Ⅱ型前列腺炎的病原谱涵盖多种革兰阳性菌和革兰阴性菌,还有CT、CMV、HSV-2和HPV。Ⅲ型前列腺炎中解脲支原体阳性率为37.0%（30/81）,沙眼衣原体阳性率为13.6%（11/81）。前列腺液病毒感染率分别为HPV8.6%（7/81）,HSV-211.1%（9/81）,CMV18.5%（15/81）。研究组前列腺液细菌16SDNA阳性率为66.7%,其中Ⅱ型前列腺炎为100%,Ⅲ型前列腺炎为61.4%。结论慢性前列腺炎患者前列腺内微生物种类繁多,其中许多微生物可能是Ⅲ型前列腺炎的致病因素。     

Estimation of Type-Specific Neutralizing Antibody to Herpes Simplex Virus Type 2 in Uterine Cervical Cancer Patients by a New Absorption Method

A simple method of estimating type-specific neutralizing antibody to type 2 herpes simplex virus (HSV-2) was devised with the use of the microneutralization system. Serially diluted serum was mixed in the well with a constant amount of type 1 virus (HSV-1), and after 3 days' incubation at 37 C, the plate was irradiated with ultraviolet light. The absorbing HSV-1 consisted of culture fluid plus an extract of infected Vero cells not especially concentrated. The well then received indicator HSV-1 or HSV-2, and after being left at 37 C for 1 hr a suspension of dispersed Vero cells was dropped into the wells, following our standard neutralization procedure. Preliminary tests with rabbit antisera showed that even a low level of HSV-2 antibody was detected by this method, unless an exceptionally high titer of HSV-1 antibody originally coexisted with the HSV-2 antibody. Sera from acutely infected persons testified to the specificity of the antibody so detected. It was revealed by means of the new technique that the rate of HSV-2 antibody was significantly higher in uterine cervical cancer patients than in control women. There was no correlation between the clinical stage of cervical cancer and the presence of HSV-2 antibody.     

正常妇女和下生殖道炎症及不孕症时泌尿生殖道中的溶脲脲原体

本文报道165例妇女宫颈阴道分泌物和部分不育男性精液的溶脲脲原体的分离结果。正常妇女组检出率为33％(11/33)、宫颈炎和阴道炎组为67％(39/59)、不孕症组80％(35/44)、富颈癌组24％(5/21)。不孕夫妇中,13例男性精液检查有9例溶脲脲原体阳性。经比较有炎症组和不孕症组的溶脲脲原体检出率高于正常组2倍多,差异显著(P<0.01和<0.005)。鉴于溶脲脲原体作为人泌尿生殖道中的常住菌,其微生态意义与下生殖道炎症和某些不孕症的关系值得进一步深入研究。     

Prevalence of serum antibodies to LA-1 oncoprotein, herpes simplex virus type-2 glycoprotein and human papillomavirus type 16 transactivator (E2) protein among Indian women with cervical neoplasia.

Prevalence of serum antibodies to synthetic peptide to oncoprotein of LA-1 known as oncogene of herpes simplex virus type-2, herpes simplex virus type-2 glycoprotein-D as an determinant of viral pathogenicity and human papillomavirus type 16 transactivator E2 protein was studied among 46 Indian women with cervical neoplasia using immunoblot assay for HSV-2 gD glycoprotein and LA-1 antibodies as well as peptide ELISA assay to detect HPV16 E2 antibodies. The seropositivity to LA-1 oncoprotein was found to be high (61%) among patients with invasive cervical carcinoma as compared to 35% in various grades of cervical intraepithelial neoplasia (CIN) and 36% in normal control women. In contrast to this, a uniformly high frequency of antibody to HPV 16 E2 was observed among women with CIN (68%), normal healthy controls (50%) and invasive cervical carcinoma (43%). However, a low frequency of seropositivity (13%) to recombinant vaccinia virus HSV-2 gD protein was found among 15 tested sera each from group of women with various grades of CIN as well as invasive cervical carcinoma as compared to 28% among seven normal healthy control. A negative correlation of LA-1 and HPV16 E2 seropositivity on patient by patient comparison among CIN and invasive cervical carcinoma group was observed which is statistically significant (P = 0.019 for CIN; P = 0.038 for invasive cervical carcinoma). However, a positive correlation (P = 0.144) was found among normal control women. The study has shown a desirable serological marker of cervical neoplasia. This serological marker could be employed as a screening tool in conjunction with cytopathological screening to diagnose women harbouring LA-1 oncogene associated cervical lesions.     

Mutagenicity testing of seminal fluid: seminal fluid increases the mutagenicity of the precursor mutagen benzo[a]pyrene in the presence of S9 mix

Small amounts of seminal fluid strongly enhanced the mutagenicity of the precursor mutagen benzo[a]pyrene (BP) in the Salmonella/microsome test. This previously unreported effect was found only in the presence of S9 mix for metabolic activation. The increase far exceeded the additive effect expected from experiments where seminal fluid and BP were tested separately with S9 mix. Testing of the direct-acting mutagen 4-nitro-o-phenylene-diamine (NPD) together with seminal fluid resulted in a lower mutagenic activity than that of NPD alone. Seminal fluid had a bactericidal effect on the Salmonella bacteria, thus only volumes up to 40 microliter could be used per plate. The mutagenic effect of only seminal fluid and S9 mix was slightly increased over controls in a standard Ames test, but was equal to the spontaneous mutation rate with a preincubation test modified according to Kado and coworkers. There were no significant differences between seminal plasma from smokers and non-smokers in any experimental series. Seminal fluid concentrated 20-fold by extraction with the mutagen-removing adsorbant Mutasorb did not have any enhancing effect on the mutagenicity of BP, nor did it exhibit any mutagenic activity in itself with or without S9 mix.