目的 研究浙江省丽水地区妇女X-射线交错互补修复基因1(XRCC1)Arg194Trp、Arg280His和Arg399Gln单核苷酸多态性与人乳头瘤病毒(HPV)(16、18、52、58)感染宫颈鳞状细胞癌发病风险的关系.方法 采用实时荧光聚合酶链式反应方法检测2016年5月-2019年7月浙江省丽水市中心医院收治的115例HPV(16、18、52、58)宫颈鳞状细胞癌患者和143例健康对照者血液标本的Arg194Trp、Arg280His和Arg399Gln三个SNP基因型频率分布,分析其与HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险的关系,28种HPV基因型采用聚合酶链式反应-反向点杂交法对宫颈分泌物标本进行检测.结果 宫颈鳞状细胞癌组和对照组XRCC1-Arg194Trp、Arg280His和Arg399Gln基因型频率分布均符合Hardy-Weinberg平衡定律(P>0.05),宫颈鳞状细胞癌组和对照组XRCC1-Arg194Trp和Arg399Gln基因型和等位基因频率分布差异有统计学意义(P<0.05),宫颈鳞状细胞癌组和对照组XRCC1-Arg280His基因型和等位基因频率分布差异无统计学意义(P>0.05),XRCC1-Arg194Trp多态性位点中,携带Trp基因的个体HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险是Arg个体的1.793倍(OR=1.793,95％CI=1.190～2.702,P<0.05);XR-CC1-Arg399Gln多态性位点中,携带Gln基因的个体HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险是携带Arg基因个体的1.668倍(OR=1.668,95％CI=1.126～2.472,P<0.05);而XRCC1-Arg280His的多态性位点与HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险无关(OR=1.210,95％CI=0.753～1.945,P>0.05).结论 浙江省丽水地区XRCC1-Arg194Trp、Arg399Gln多态性可能与人群HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险相关,而XRCC1-Arg280His多态性与本地区人群HPV(16、18、52、58)感染宫颈鳞状细胞癌易感性无关.

目的 分析浙江省丽水地区人群亚甲基四氢叶酸还原酶(MTHFR)基因C677T和A1298C单核苷酸多态性与人乳头瘤病毒(16、18、52、58)感染的宫颈鳞状细胞癌发病风险的关系.方法 采用实时荧光聚合酶链式反应方法检测在丽水市中心医院住院治疗的115例HPV(16、18、52、58)感染的宫颈鳞状细胞癌患者和143例HPV阴性的健康对照妇女血液标本MTHFR C677T和A1298C两个单核苷酸多态性基因型频率的分布,分析其与HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险的关系;28种HPV基因型采用聚合酶链式反应－反向点杂交法对宫颈分泌物标本进行检测.结果 病例组和对照组MTHFR C677T和A1298C基因型频率分布均符合Hardy-Weinberg平衡定律(均P＞0.05),病例组和对照组MTHFR C677T和A1298C基因型频率分布均有统计学差异(均P＜0.05).在MTHFR C677T多态性位点中,携带T的受试者HPV(16、18、52、58)感染的宫颈鳞状细胞癌发病风险是C受试者的1.871倍(OR=1.871,95％CI:1.294～2.705,P＜0.05);在MTHFR A1298C多态性位点中,携带C的受试者HPV(16、18、52、58)感染宫颈鳞状细胞癌发病风险是A受试者的1.867倍(OR=1.867,95％CI:1.204～2.896,P＜0.05).结论 MTHFR C677T和A1298C多态性可能与浙江省丽水地区人群HPV(16、18、52、58)感染者宫颈鳞状细胞癌发病有关.

OBJECTIVE To explore the relationship between virulence genes in clinical isolates of Ureaplasma urealyticum (Uu) and pathogenicity so as to clarify the genetic background of pathogenicity.METHODS A total of 2661 specimens were collected from patients with Uu infections who were treated in hospitals from Dec 2012 to Dec 2016,totally 201 strains of highly pathogenic Uu were screened and isolated,which were isolated from female patients with infertility,premature rupture of fetal membranes,threatened abortion and urinary system disease.The downstream primers of 4 conservative housekeeping genes of Uu (ftsH,rpL22,valS,thrS) and 2 variable virulence genes (ureG and rnbanp 1) were designed,and the 6 genes were detected and sequenced by using polymerasechain-reaction (PCR).RESULTS Totally 201 clinical isolates of highly pathogenic Uu were amplified by PCR,all of the amplified products were target genes through the sequencing,and the positive rates of ureG and mbanp 1 were 93.1％ and 90.6％,respectively.The result of drug susceptibility testing showed that the drug resistance rate of the 201 strains of highly pathogenic Uu was higher than that of the strains isolated from overall population,the drug resistance rates to levofloxacin,roxithromycin and azithromycin were the highest,which were respectively 21.2 ％vs15.2％,12.5％vs6.9％ and 10.9％vs3.5％.The positive rate of Uu infection tended to be same in the hospital from 2012 to 2016 and showed downward trend in recent two years.CONCLUSION Most of the 201 strains of Uu carry with the ureG and mbanpl genes,showing that the highly pathogenic clinical isolates have the pathogenic genes in common.The positive rate of Uu infection does not show a sign of significant rise in this area in recent years,however,the drug resistance rate of the 201 strains of highly pathogenic Uu is remarkably higher than that of the common strains.It is necessary for the clinicians to strengthen the identification of the pathogenic Uu strains and reasonably use antibiotics based on the result of drug susceptibility testing so as to prevent the increase of drug resistance rate of the clinical isolates of Uu.

目的:了解丽水地区妇女宫颈人乳头瘤病毒(HPV)感染的阳性率与各年龄组关系及亚型分布情况,为该地区宫颈癌防治及流行病学研究提供依据.方法:采用DNA反向斑点杂交技术对2 156名妇女宫颈脱落细胞标本进行21种HPV基因型检测,统计分析HPV感染的阳性率和年龄组的关系及亚型分布特点.结果:2 156例标本中阳性944例,阳性率为43.8％.检出前8位亚型优势型别依次为16、58、52、66、53、68、51、33.HPV感染的高峰年龄是30～50岁,检出率最高的年龄组是＞60岁的人群,其次是50～60岁年龄组.各年龄组的HPV阳性检出率及优势亚型检出率有显著差异.结论:HPV基因分型检测对宫颈癌的防治有重要意义.

Objective To investigate the distribution of human papillomavirus (HPV) genotypes in male patients with anogenital warts in Lishui area,Zhejiang province.Methods Tissue specimens were obtained from the lesions of 150 male patients with anogenital warts.PCR-reverse dot blot hybridization was performed to detect the presence of 3 low-risk HPV types (HPV 6,11,and 43) and 16 moderate-or high-risk HPV types (HPV 16,18,31,33,35,39,45,51,52,53,56,58,59,66,68 and CP8304) in these specimens.Chi-square test was used for statistical analysis.Results HPV was detected in 91 (60.67％) of the 150 male patients.Of the 91 positive patients,74 (81.32％) were infected by single or multiple low-risk HPV types,whereas 17 (18.68％) by single or multiple high-risk HPV types.Thirty-one (34.07％) patients harbored 2-5 HPV genotypes,including 20 (64.52％)patients infected with both low-risk and high-risk HPV types,and 6 (19.35％) patients infected with two low-risk HPV types.The coexistence of moderate-or high-risk HPV types with HPV 6 was observed in 13 (41.94％)patients,and that with HPV 11 in 6 (19.35％) patients.The most prevalent genotype was HPV 6 (28.68％,39/136),followed by HPV 11 (26.47％,36/136),16 (8.09％,11/136),52 (5.15％,7/136),53 (5.15％,7/136),51 (4.41％,6/136),58 (4.41％,6/136) and 43 (4.41％,6/136).Conclusions Low-risk HPV genotypes predominate in male patients with anogenital warts,and there are large differences in the distribution of multiple infections and HPV genotypes.

尖锐湿疣(Conyloma acuminate,CA)是由人乳头瘤病毒(Human papilloma virus,HPV)感染所致,在我国是最常见的性病之一,发病率居淋病之后,占第二位,且发病率逐年上升.由于有大量的尖锐湿疣病例未报告,所以实际发病率还要更高.该病常以亚临床感染形式存在,传染性强,复发率高,临床治疗较为棘手.CA病例的HPV流行型别有地区差异性,不同型别的HPV致病性有差别.为了了解丽水地区CA组织中HPV的感染型别,对丽水市中心医院2012年1-11月共129例患者CA组织中的HPV进行检测及分型,结果报告如下.

Objective To investigate the distribution of hepatitis B virus(HBV) genotypes in She population compared with that in Han population also in Lishui area,and to further study the liver injuries and HBV serum markers among She populations infected with HBV of different genotypes.Methods The genotypes of HBV DNA positive subjects were examined by real time fluorescence polymerize chain reaction and S gene sequencing,while ALT and AST by automatic biochemical analyzer and HBV serum markers by ELISA.Results 173 HBV DNA positive samples were screened from 1744 She population,and 78 were genotype B,92 genotype C,3 genotype B/C.The mean HBV DNA copy logarithm of subjects infected with HBV genotype B and C were 4.596±1.814 and 5.364±2.189 respectively.13 of those infected with HBV genotype B and 15 of genotype C were detected with enhanced ALT and AST;and 11 of genotype B and 26 of genotype C were HBeAg positive.The same number of Han populations were also examined as contrasts.Conclusions It can be concluded that She population in Lishui area is mainly infected with HBV genotype B and C,and genotype C takes more proportion,which is obviously different from Han population in the same area.For She population in Lishui,the HBV DNA copy and HBeAg positive rate of HBV genotype B are lower than those of HBV genotype C,which indicates that populations infected with HBV genotype B might have less active virus,lighter liver injuries and better prognosis.

Objective To investigate the infection of Human Papillomavirus(HPV) genotype in cervical diseases and their distribution characteristic in women in Llishui area.Methods The clinical common genotype and the characteristic distribution of HPV was analyzed.A total of 2638 women patients in the locality during 19,Sep.2008 to 15,Mar.2011 were selected for 21 genotypes(6,11,16,18,31,33,35,39,42,43,44,45,51,52,53,56,58,59,66,68,CP8304)HPV detecting with HybriMax.Results The total positive rate of HPV infection was 50.87%.The 10 most common high-risk genotypes were HPV16,52,58,33,31,53,68,CP8304,18,39.The 2 most common low-risk genotypes were HPV11,6.The total multiple infection rate was 32.41 % and two types co-infection were the most common situation.Conclusion The prevalence of HPV infection and multiple infection are common.The infection genotypes are accordant to those of Asia,yet still have its own characters.

为了解丽水地区畲族人群乙型肝炎病毒(hepatitis B virus,HBV)基因型感染分布,本调查共筛查1 744份畲族样本,分析畲族人群乙型肝炎病毒基因型分布状况,并与本地区汉族人群[1]进行初步比较,现将结果报告如下.

目的:分析丽水市中心医院鲍曼不动杆菌耐药特征及blaOXA23基因携带情况,指导临床合理应用抗生素.方法:经VITEK2-compact微生物系统鉴定,收集丽水市中心医院2008年3月至2009年6月从临床分离的鲍曼不动杆菌62株.统计其科室来源,以K-B法检测菌株对抗生素的耐药性,并利用PCR扩增法检碳青酶烯酶耐药基因blaOXA23.结果:鲍曼不动杆菌检出率前两位是脑外科和ICU病房,分别占32.3%和27.4%.多重耐药鲍曼不动杆菌阳性率最高的是ICU,为70.6%(12/17);其次为脑外科,为35.0%(7/20).亚胺培南、美洛培南、头孢他啶、头孢吡肟、替卡西林/克拉维酸、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦、多粘菌素耐药率分别为33.9%、38.9%、61.3%、53.3%、56.1%、45.2%、16.9%和0.0%.blaOXA23总检出率35.5%,亚胺培南耐药鲍曼blaOXA23检出率100.0%,敏感鲍曼均未检出.结论:鲍曼不动杆菌分布广泛,并且多重耐药性严重;blaOXA23基因可能是耐碳青酶烯类抗生素的主要原因.建议加强鲍曼不动杆菌耐药性监测及耐药机制的研究.

根据乙型肝炎病毒(HBV)全基因序列的差异≥8%或S基因序列≥4%,可以将HBV分为A～H等8种基因型.不同基因型致病性及治疗效应不同.为了解丽水市畲族人群乙型肝炎病毒基因型的分布特点及其与混居汉族人群乙型肝炎病毒基因型分布是否存在差异,我们随机抽样检测了1744例畲族及混居的557例汉族人群血样,对其中的HBVDNA阳性者采用荧光定量PCR法(试剂由深圳匹基生物工程有限公司提供)进行病毒基因分型检测,现将结果报告如下.

目的总结缺血性肠病的临床特点,提高对该疾病诊治的认识。方法收集缺血性肠病患者23例,对其临床特征进行回顾性分析。结果主要临床表现为腹痛、腹泻、便血、恶心、呕吐及发热等。腹痛23例,腹泻11例,便血17例。所有患者均行彩色多普勒超声检查,阳性率43.5%(10/23);17例行全腹螺旋CT检查,阳性率52.9%(9/17);肠系膜上、下动脉造影阳性率100%(8/8),造影检查发现血管畸形、管腔狭窄、闭塞;1例腹部立位X线检查提示肠梗阻。所有患者经恰当治疗后均痊愈出院。结论缺血性肠病临床症状与体征无特异性,误诊率高,如及时行肠镜等辅助检查能早期发现,及早治疗,多数患者预后良好。

为了解丽水市畲族人群乙型肝炎病毒基因型的分布特点及其与HBeAg关系,为临床提供帮助,我们随机抽样检测了1744例畲族人群标本,对其中的HBV DNA阳性者进行了病毒基因分型及HBeAg检测,现报道如下.

目的了解丽水地区人乳头状瘤病毒(HPV)基因型在宫颈疾病中的感染状况及其分布特点。方法运用HybriMax医用核酸分子快速杂交系统对该地区288例妇科患者进行21种HPV(6、11、16、18、31、33、35、39、42、43、44、45、51、52、53、56、58、59、66、68、CP8304)基因型的分型检测,分析常见感染亚型和分布特点。结果研究人群HPV总阳性率为47.6%,常见的10种高危型为HPV16、58、52、33、53、31、66、39,CP8304、51;常见的2种低危型为HPV11、6。多重感染的比例为35.8%,其中以二重感染最多见。结论该地区HPV感染率和多重感染比例均较高,常见感染亚型总体符合亚洲人群的分布规律,又有独特的区域分布特点,值得重视。

为了解丽水地区汉族农村人群中乙型肝炎病毒(HBV)基因型的分布,我们共筛查2 478份样品,对其中HBV DNA呈阳性的乙型肝炎病人进行病毒基因型分析,现将结果报告如下.

从老年慢性阻塞性肺疾病患者痰标本中分离到1株罕见的泛耐药黏液型鲍曼不动杆菌(slime-production pandmg-re-sistant A. baumannii,SPAB),现报告如下.

宫颈癌为最常见的妇科恶性肿瘤,在全球妇女癌症病死率中位居第二,近年来多项研究结果已证实,人乳头状瘤病毒(HPV)持续感染为其主要病因[1-2],因此目前HPV感染检测已成为宫颈癌中必不可少的筛查内容.根据HPV不同基因型与宫颈癌发生的危险性高低可分为低危型HPV和高危型HPV,前者常可引起外生殖器湿疣、宫颈上皮内低度瘤变等良性病变,而后者则与宫颈癌和宫颈上皮内高度瘤变的发生密切相关[3].

Objective:To establish a real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)to detect FOXP3 mRNA expression in human peripheral blood mononuclear cells(PBMCs)and to investigate the correlations between activity of CD4+CD25+ regulatory T cells and expression of FOXP3 mRNA.Methods:Total RNA was extracted with Trizol from human PBMCs and mRNA was transcribed reversely into cDNA.Real-time fluorescent quantitative RT-PCR with β-actin as the internal control gene was used to detect the expression levels of FOXP3 mRNA in 29 pediatric patients with asthma and 24 age-matched healthy children.The specificity of PCR productions was identified with the dissociation curves and agarose gel electrophoresis.Flow cytometry analysis was used to assess the percentage of CD4+CD25+regulatory T cells.The correlation between the expression and activity was analyzed.Results:The percentages of CD4+CD25+ regulatory T cells in group of asthma were significantly lower than those in group of control(P0.01),so did the expressions of FOXP3 mRNA(P0.05).The analysis of dissociation curves on FOXP3 and β-actin showed that both of them had only one simple spike and the Tm values were 82.4 ℃ and 87.8 ℃,respectively.The agarose gel electrophoresis of them showed only single PCR product each.Conclusion:It is an easy and reliable test to detect the expression level of FOXP3 mRNA by SYBR GreenⅠreal-time fluorescence quantitative RT-PCR and the preliminary experimental result shows that the percentages of CD4+CD25+ regulatory T cells and the expressions of FOXP3 mRNA have the same trend and confirms the correlation between them.

目的探讨人外周血单个核细胞中FOXP3mRNA的表达与CD4^＋CD25^＋Treg表达的相关性。方法提取人外周血单个核细胞总RNA,逆转录成mRNA,以β-actin为内参照,实时荧光定量RT-PCR检测29例哮喘患儿及24例同龄对照FOXP3mRNA的相对表达量,采用融解曲线和琼脂糖电泳鉴定PCR产物特异性;同时采用流式细胞技术检测CD4^＋CD25^＋Treg的比例。结果哮喘患儿组CD4^＋CD25^＋Treg细胞百分率明显低于同龄对照组,P〈0.01;FOXP3mRNA的表达也显著降低,P〈0.05;FOXP3和β-actin的融解曲线分析表明均仅有单一峰,Tm值分别为82.4℃和87.8℃;琼脂糖电泳显示都仅有单一扩增产物。结论应用SYBRGreenⅠ实时荧光定量RT-PCR技术检测FOXP3mRNA表达水平简便、经济、可行,哮喘患儿CD4^＋CD25^＋Treg明显降低可能参与哮喘发病,FOXP3表达降低可能是导致CD4^＋CD25^＋Treg发育障碍的重要因素。

Objective To study the significance of the correlation between FOXP3 mRNA in mononuclear cells and CD4+CD25+Treg. Methods Total RNA was extracted with Trizol from human peripheral blood mononuclear cells(PBMC) and mRNA was transcribed reversely into cDNA. The real-time fluorescence quantitative RT-PCR was used to detect the expression of FOXP3 mRNA in 29 pediatric patients with asthma and 24 healthy children with β-actin as the internal control gene. The specificity of FOXP3 and β-actin amplifications was confirmed by analyzing the dissociation curves and electrophoresis of the target amplicon. Flow cytometry analysis was performed to assess the percentage of CD4+CD25+ regulatory T cells. Results The percentage of CD4+CD25+ regulatory T cells and FOXP3 mRNA expression of the asthma children was significantly lower than that of the healthy children(P0.01, P0.05) respectively. The specificities of FOXP3 and β-actin amplifications were confirmed by analyzing the dissociation curves and electrophoresis of PCR products with the values of Tm 82.4℃ and 87.8℃, respectively. Conclusions Quantitative detection of FOXP3 mRNA by the method of SYBR GreenⅠreal-time fluorescence quantitative RT-PCR is stable and reliable. The decrease of CD4+CD25+ regulatory T cells may be involved in the pathogenesis of asthma. The decreased mRNA expression of FOXP3 may be associated with the aberrant development of CD4+CD25+ regulatory T cells.