男性尖锐湿疣患者尿样中人乳头瘤病毒检测的临床意义

目的　探讨男性尖锐湿疣患者尿样中人乳头瘤病毒检测的临床意义。方法　用 PCR方法对 70份尿样和 5 2份石腊包埋组织进行 HPV DNA检测。结果　 5 0例尿道内尖锐湿疣患者中有 4 3例 (4 3/ 5 0 ,86 % ) HPV DNA阳性 ,7例尿道内尖锐湿疣患者 (7/ 5 0 ,1 4 % )、2例包皮系带处尖锐湿疣和 1 8例正常男性对照尿样为阴性 ;5 2例石腊包埋组织 HPV DNA均阳性 ;4 3例尿中 HPV DNA的阳性类型与其相对应的石腊包埋组织 HPV DNA完全一致。结论　对男性无症状的生殖道 HPV感染 ,尿样是一种非创伤性检查 ;尿样 HPV DNA检测有助于男性尿道内尖锐湿疣治疗效果的监测     

Epoxide hydrase in human liver biopsy specimens: assay and properties

A sensitive assay for the determination of epoxide hydrase activity in needle biopsy specimens of human liver has been developed with [³H]styrene oxide as substrate and was used for the study of some properties of human epoxide hydrase. Levels of epoxide hydrase in liver of man are 4.71 ± 0.41 nmoles styrene glycol/mg protein per min, comparable to guinea-pig (5·00 ± 0·38) rather than to Rhesus monkey (13·16 ± 0·88). Human hepatic epoxide hydrase which was found exclusively in the microsomal fraction, was solubilized with Cutscum, a neutral detergent, and purified. The enzyme was remarkably stable as long as it was particle-bound. Neither dialysable cofactors nor endogenous activators or inhibitors appeared to exist in the homogenate. The optimum pH for the purified enzyme was 9. The apparent K_m was 0.38 mM and the apparent V_max was 62.1 nmoles product/mg protein per min with respect to styrene oxide as the substrate. The product, styrene glycol, had no inhibitory effects. High (5–17 mM) concentrations of substrate, styrene oxide, markedly inhibited epoxide hydrase activity at low (0·2 mg/ml) but not at high (2 mg/ml) concentrations of protein, indicating that this inhibition may be due to the alkylating properties of the subsirate, styrene oxide. Sulfhydryl reagents slightly but significantly inhibited the enzyme suggesting that no sulfhydryl group is essentially involved in the catalytic mechanism at the active site, but that sulfhydryl group(s) may be of importance for holding the enzyme molecule in the optimal conformation for maximal activity. Chelating agents, a carbonyl reagent and β-diethylaminoethyl diphenylpropylacetate (SKF 525-A) had no effect.     

Monooxygenase activity of human liver in microsomal fractions of needle biopsy specimens.

1 Methods are described for the determination of mixed function oxidase activity in microsomal fractions from percutaneous needle biopsies of human liver. 2 Activities of needle biopsy samples were comparable with those of wedge biopsy sample obtained at laparotomy from different subjects. 3 Although cytochrome P-450 content of liver from rat and man was similar, human AHH activity was only 10% of that in the rat. 4 In biopsies with preserved hepatic architecture, AHH activity and cytochrome P-450 content showed a significant positive correlation. 5 Cigarette smoking significantly increased both AHH activity and turnover number, but not cytochrome P-450 content, of biopsies with normal architecture. 6 The presence of liver disease caused a significant decrease in AHH activity and cytochrome P-450 content.     

The start of a programme for measuring diamine oxidase activity in biopsy specimens of human renal mucosa

In human subjects, apart from in the kidney, diamine oxidase occurs mainly in the gut. Therefore this enzyme can be used as an indicator of intestinal integrity. In biopsies of rectal mucosa the diamine oxidase activity was assayed in 55 patients, 41 having a histologically normal mucosa and 14 being diseased. The determinations of the enzymic activity were supervised by statistical quality control. In the unchanged rectal mucosa the diamine oxidase activity was 40 nmol/min X g on average. In 7 patients with rectal polyps the enzymic activity was significantly diminished in these benign tumors (mean = 7.7 nmol/min X g) apart from one, where it was elevated. A decrease in diamine oxidase activity was further observed in rectal carcinoma and ulcerative colitis. Whether the reduction of intestinal diamine oxidase activity accompanies premalignant or malignant states or whether it is a general sign of a disturbance of intestinal integrity remains questionable.     

Frequency distribution of dextromethorphan O-demethylation in a Greek population

OBJECTIVE: To determine the CYP2D6 phenotype in a Greek population by using dextromethorphan (DM) as a probe drug. METHODS: DM (30 mg) was given orally to 102 unrelated Greek subjects and 8-hour urine samples were collected. Concentrations of DM and its metabolite dextrorphan (DX) were determined using a validated HPLC assay. Metabolic molar ratio (MR) of DM to free DX in log form was used as an in vivo index of metabolic status. RESULTS: The frequency distribution histogram of MR was bimodal. An antimode of 0.25 for the mean log MR was determined using probit analysis. Seven of 102 subjects (6.9%) were poor metabolizers (PMs). CONCLUSION: The PM frequency of CYP2D6 in Greek subjects was similar to other Caucasian populations.     

流动人口密集地区宫颈上皮内瘤变与高危型人乳头瘤病毒关系调查

目的 了解流动人口密集地区宫颈上皮内瘤变与高危型人乳头瘤病毒间的关系及相关因素.方法 对妇科门诊所有感染HPV16、HPV18、HPV31亚型患者的基本资料进行初步登记,再通过查阅电子病历或电话回访逐一完善相关资料.结果 从198例高危HPV感染患者中检出的217频次的阳性结果中发现：HPV16感染比率最多,占61.75％,HPV18其次,占25.35％,HPV31位居第三,占12.90％.198例患者中过早或多伴侣性生活的占16.16％,多孕、多产和多次人流占32.83％、感染性阴道炎占90.40％、慢性宫颈炎占81.82％;在198例高危HPV感染者中共发现了宫颈上皮内瘤变及宫颈浸润癌26例,26例宫颈上皮内瘤变及宫颈浸润癌患者中22例感染HPV16,其恶性程度也更高.结论 流动人口密集地区宫颈上皮内瘤变与高危型HPV16感染密切相关,慢性宫颈炎及阴道感染所致宫颈低免疫力是最主要的高危HPV感染因素.     

高危型人乳头瘤病毒DNA载量在宫颈病变中的应用

目的探讨高危型人乳头瘤病毒（HR-HPV）的载量在宫颈病变诊断中的应用价值。方法246例21～69岁妇女采用第2代杂交捕获试验（HC2）进行HPV DNA检测,病毒载量由样本的相对光单位（RLU）与标准阳性对照（PC）之比来衡量。根据HR-HPV检测值将RLU/PC分为阴性（〈1）、低度载量（1～9）、中度载量（10～99）和高度载量（≥100）。宫颈病变按照病理诊断分为宫颈炎、CINⅠ级、CINⅡ～Ⅲ级、宫颈癌（SCC）。结果随着宫颈病变程度发展,中、高度载量病例所占比例增加（宫颈炎组30.61%,CINⅠ组83.33%,CINⅡ～Ⅲ组90.48%,SCC组100%）。结论宫颈HR-HPV病毒载量是影响宫颈病变及发展的重要因素。     

人乳头瘤病毒基因分型检测联合液基薄层细胞学检查的临床应用

目的研究并探讨人乳头瘤病毒(HPV)基因分型检测联合液基薄层细胞学检测(TCT)在宫颈癌筛查中的应用价值。方法于2015年5月~2016年6月,选取120例疑似宫颈癌患者作为此次研究的对象,所有患者均接受HPV检测、TCT检查,以病理诊断结果作为金标准,计算HPV、TCT以及HPV+TCT对宫颈癌的诊断敏感性、特异性、准确性。根据宫颈病变程度进行分组,将患者分为CINⅠ级、CINⅡ级、CINⅢ级、宫颈癌,观察不同宫颈病变程度的HPV病毒载量,并计算宫颈病变程度与HPV病毒载量之间的相关性。结果 HPV的敏感性明显高于TCT(P0.05),TCT的特异性明显高于HPV(P0.05),两种检测方法的准确性相当(P0.05);HPV+TCT的诊断准确性为95.83%,较HPV、TCT更高(P0.05)。宫颈病变程度与HPV病毒载量密切相关(P0.05),且呈正相关。结论在宫颈癌的筛查中,HPV与TCT联合诊断的准确性高,可减少漏诊、误诊。     

Detection of human papillomavirus DNA, p53 and ki67 expression in penile carcinomas

Our study is aimed at evaluating the presence of p53 and Ki67 expression by immunohistochemistry in a series of 11 paraffin-embedded penile carcinomas. We also investigated the presence of Human Papillomavirus (HPV) DNA in these tumours and performed an accurate typing by DNA sequencing on positive samples. Immunohistochemistry (IHC) was performed with the anti-p53 and Ki67 mouse monoclonal antibodies. DNA extracted from small sections of each specimen was submitted to amplification with HPV specific general primers; PCR products of the proper length were purified and sequenced. IHC demonstrated nuclear accumulation of mutated p53 and Ki 67 expression in 10/11 tumour samples (90.9%). The prevalence of HPV DNA was 72.7%; the most prevalent type was HPV16. Sequencing analysis revealed the presence of HPV53 (12.5%), HPV18 (25%) and HPV16 (62.5%). Out of the p53 or Ki67 positive carcinomas the percentage of HPV positives was 80% and 70% respectively. Our results indicate that penile carcinoma is frequently associated to high risk HPV and with diffuse p53 and Ki67 expression.     

采用基因分型质控品评价荧光PCR法人乳头瘤病毒核酸检测试剂盒

目的:使用人乳头瘤病毒(HPV)L1基因分型质控品对荧光PCR法人乳头瘤病毒核酸检测试剂盒进行评价。方法:以中国药品生物制品检定所体外诊断试剂与培养基室建立的含有30种不同型别HPV基因分型质控品盘为样本,按照各个试剂盒的要求进行检测。结果:使用含有30种不同型别HPV基因分型质控品盘为模板进行试剂盒评价时,A试剂检测有23种型别检测结果符合,7种型别不符合,HPV 59型漏检,HPV 53,54,61,66,72,81型有交叉现象。B试剂检测有29种型别检测结果符合,1种型别不符合,HPV 58型漏检。C试剂检测有26种型别检测结果符合,4种型别不符合,HPV 43,66,81,CP 8304型有交叉现象。D试剂有29种型别检测结果符合,1种型别不符合,HPV 68型漏检。对照检测E试剂30种型别的检测结果全部符合。4家公司的试剂基本能正确区分质控品中HPV基因型,但有部分的HPV基因型有漏检现象和交叉反应。对照试剂无漏检交叉现象。结论:含有30种不同型别的HPV-L1基因分型质控品能准确地评价4家公司的HPV核酸检测试剂盒(荧光PCR法)和高危型人乳头瘤病毒DNA检测试剂盒(酶切信号放大法)的性能;在HP...     

Biflorin induces cytotoxicity by DNA interaction in genetically different human melanoma cell lines

Cancer is a public health problem and the second leading cause of death worldwide. The incidence of cutaneous melanoma has been notably increasing, resulting in high aggressiveness and poor survival rates. Taking into account the antitumor activity of biflorin, a substance isolated from Capraria biflora L. roots that is cytotoxic in vitro and in vivo, this study aimed to demonstrate the action of biflorin against three established human melanoma cell lines that recapitulate the molecular landscape of the disease in terms of genetic alterations and mutations, such as the TP53, NRAS and BRAF genes. The results presented here indicate that biflorin reduces the viability of melanoma cell lines by DNA interactions. Biflorin causes single and double DNA strand breaks, consequently inhibiting cell cycle progression, replication and DNA repair and promoting apoptosis. Our data suggest that biflorin could be considered as a future therapeutic option for managing melanoma.     

Recent developments in human papillomavirus vaccines

The association of certain high-risk human papillomaviruses with the development of anogenital cancer in humans is well-established. Numerous preclinical studies have underwritten the development of both prophylactic and therapeutic vaccine candidates for clinical evaluation. Prophylactic strategies are utilising virus-like particles composed of the L1 viral capsid protein to induce neutralising antibodies while therapeutic approaches are aimed at generating specific T cells targeted at the viral E6 and/or E7 oncogene products. Thus far, human papillomavirus virus-like particle vaccines have proven to be clinically efficacious in the early trials looking at the prevention of infection. Important future milestones will be showing the prevention of high-grade cervical intraepithelial neoplasia and sufficient longevity for such protection. Different types of therapeutic vaccines including peptide, protein, DNA or viral vector-based vaccines have proven to be safe and immunogenic in patients, although there is often no correlation with clinical outcome. The possibility of combined prophylactic and therapeutic vaccines may offer the best chance for a significant reduction in the incidence of death from cervical cancer worldwide.     

Recent developments in human papillomavirus vaccines

The association of certain high-risk human papillomaviruses with the development of anogenital cancer in humans is well-established. Numerous preclinical studies have underwritten the development of both prophylactic and therapeutic vaccine candidates for clinical evaluation. Prophylactic strategies are utilising virus-like particles composed of the L1 viral capsid protein to induce neutralising antibodies while therapeutic approaches are aimed at generating specific T cells targeted at the viral E6 and/or E7 oncogene products. Thus far, human papillomavirus virus-like particle vaccines have proven to be clinically efficacious in the early trials looking at the prevention of infection. Important future milestones will be showing the prevention of high-grade cervical intraepithelial neoplasia and sufficient longevity for such protection. Different types of therapeutic vaccines including peptide, protein, DNA or viral vector-based vaccines have proven to be safe and immunogenic in patients, although there is often no correlation with clinical outcome. The possibility of combined prophylactic and therapeutic vaccines may offer the best chance for a significant reduction in the incidence of death from cervical cancer worldwide.     

Sociodemographic correlates of abstinence and excessive drinking in the Greek population

The objective of this study was to investigate the determinants of abstinence and drinking patterns among 14,367 Greeks. Participants older than 60, single and widowed men, as well as participants living outside the Greater Athens area were significantly more likely to abstain than to be "light to moderate" drinkers. In contrast, those at higher educational levels and those separated or divorced were less likely to abstain than their counterparts. With respect to excessive drinking, older people and those at higher educational levels were less likely to engage excessive drinking than their counterparts. Among men, subjects who were separated or divorced were more likely to drink excessively than those who were married. The results of our study help detect groups at high risk for diseases associated with high or low consumption of alcohol, and outline preventive strategies.     

人乳头瘤病毒流行病学研究进展

人乳头瘤病毒(human papillomavirus,HPV)持续感染是妇女发生宫颈癌的必要因素,此文介绍了HPV的流行病学特征,包括HPV的分布、感染的危险因素以及导致宫颈癌的流行病学证据.     

Drug population in one thousand geographically distributed urine specimens

Hundreds of drugs and other substances are excreted in urine. In a comprehensive drug screen, it is impossible to identify every detectable substance. In order to delineate the most commonly detected drugs, approximately 1000 urine specimens from geographically distributed clinical laboratories were analyzed for drug substances. Twenty-nine (29) clinical laboratories submitted up to 76 consecutive urine specimens found to be positive for substances other than nicotine and caffeine. Specimens were analyzed by thin layer chromatography and special procedures for salicylates, benzoylecgonine, PCP, benzodiazepines, and cannabinoid metabolites. Every specimen was also tested for opiates, PCP, and cannabinoid metabolites by an enzyme immunoassay procedure. The total number of drugs detected in 1000 specimens was 3014, an average of three drugs per specimen; and 110 different drugs were identified. Of these, 50 drugs accounted for 95% of the total detected; 21% of the specimens contained cannabinoid metabolites, and 4% of the specimens contained cocaine and/or benzoylecgonine. Most of the specimens were routine, and only 4% originated from comatose patients.     

应用Luminex液相芯片技术检测宫颈癌组织中人乳头瘤病毒基因型

目的：评价Luminex液相芯片技术在宫颈癌组织中人乳头瘤病毒（ HPV）基因型检测中的临床价值。方法收集20例临床确诊宫颈癌病例的石蜡组织标本,采用液相芯片法对所有标本进行HPV基因分型。考虑到HPV16亚型在宫颈癌组织中检出率较高,设计针对HPV16 E6区通用型引物采用PCR方法对其进行HPV DNA检测,然后进行测序分析。结果在20例组织病理学确诊为宫颈癌组织中,液相芯片和PCR法HPV检测率均为90%（18/20）。所有液相芯片检出的单一型感染经基因测序验证准确率为94.4%（17/18）。结论 Luminex液相芯片技术可用于检测人组织中HPV基因型,是HPV分型检测及大规模筛查的理想方法。