HPV16 E2基因各区域缺失与宫颈病变相关性的研究

目的:检测湖北地区HPV16阳性宫颈标本E2基因碳端(C)、铰链区(H)、氮端(N)的缺失状态,探讨其与CIN及宫颈癌的关系。方法:将122例宫颈标本按病变程度分为宫颈癌组、CINⅡ~Ⅲ组、对照组。应用多重聚合酶链反应(PCR)将各组HPV16感染标本E2基因的C、H、N分别与E6基因同时扩增,检测E2基因各区域的缺失状态,并应用Scion Image4.0软件半定量分析E2、E6基因条带灰度值,判断HPV DNA的整合状态。结果:宫颈癌组E2基因C、H、N缺失率分别是22.5%,50%,77.5%;CIN组为30.77%,69.23%,92.31%;对照组为9.09%,27.27%,27.27%。E2基因C、H缺失率在3组的任两组间比较无统计学差异(P0.05),而E2基因N端缺失率在宫颈癌组与对照组之间及CIN组与对照组之间差异有统计学意义(P均0.01)。结论:HPV16病毒E2基因N端缺失状态与CIN及宫颈癌有内在关系。在检测HPV感染同时检测E2基因N端缺失状态,对间接判断预后及指导后续治疗有重要意义。     

HPV16 E6基因和蛋白水平与维汉宫颈病变进展的关系研究

目的:评价HPV16 DNA E6载量和蛋白表达与宫颈病变病情进展的关系,并比较维、汉之间的差异性。方法:通过收集妇科门诊就诊的HPV16感染的维吾尔族和汉族各个级别宫颈病变妇女的宫颈组织和脱离细胞。采用实时荧光定量PCR测定HPV16 DNA E6载量,免疫组化法检测HPV16 E6蛋白表达。结果:汉族和维吾尔族中,炎症组、CIN1、CIN2~3、鳞癌组4组的HPV16 E6 DNA载量比较,差异均有统计学意义(H=30.33,P=0.001;H=37.37,P=0.001)。CIN2~3组中,维吾尔族的HPV16 E6 DNA载量高于汉族,差异有统计学意义(Z=-2.266,P=0.023)。汉族和维吾尔族中,炎症组、CIN1、CIN2~3、鳞癌组4组的HPV16 E6蛋白表达比较,差异有统计学意义(χ~2=38.85,P=0.001;χ~2=44.71,P=0.001);HPV16 E6蛋白表达随着病变程度的增加呈上升趋势。炎症、CIN1、CIN2~3中,HPV16 E6 DNA载量和蛋白表达具有明显的正相关性(P0.05);宫颈癌中HPV16 E6 DNA载量和蛋白表达无相关性(P0.05)。结论:维吾尔族CIN2~3患者的HPV16 E6 DNA载量高于汉族CIN2~3患者,E6基因在维吾尔族宫颈癌发生可能有更重要作用,HPV16 E6 DNA载量可成为预测和诊断宫颈癌前病变和宫颈癌的标志物。     

HPV16基因整合与FHIT蛋白缺失在宫颈癌变中的作用

目的 探讨HPV16的基因型整合状态及FHIT基因缺失在宫颈癌发生发展中的作用及相关性。方法选取44例宫颈癌、58例宫颈上皮内瘤样病变(CIN)和20例宫颈正常组织的患者,免疫组化检测宫颈FHIT蛋白的表达;并用多重PCR法检测HPV16 E2/E7表达。结果 122例单一HPV16阳性标本HPV16总整合率为78.69%,在正常宫颈组织、CIN1、CIN2、CIN3和宫颈癌中整合率分别为60.00%、66.67%、75.00%、86.96%和88.63%,随病变加重,整合率增强,组间差异有统计学意义(P0.05);FHIT蛋白的总缺失率为43.44%,FHIT蛋白在正常宫颈组织、CIN1、CIN2、CIN3和宫颈癌中的缺失率分别为15.00%、20.00%、25.00%、43.48%和72.73%。FHIT蛋白的缺失率,随病变加重,缺失率增高,组间差异有统计学意义(P0.05)。FHIT蛋白缺失在不同HPV16整合时不同,差异有统计学意义(P0.05)。结论 HPV16基因整合和FHIT蛋白缺失在宫颈癌的发生、发展中起重要作用,HPV16基因整合可能通过诱导FHIT基因低表达从而促使宫颈癌发生发展。     

宫颈癌中抑癌基因FHIT表达与HPV16基因状态

目的:探讨抑癌基因FHIT表达及人乳头瘤病毒16(HPV16)的基因型整合状态在宫颈癌发生发展中的作用及相关性。方法:选取柳州市人民医院2013年6月至2014年12月收治的42例宫颈癌、55例宫颈上皮内瘤样病变(CIN)和20例宫颈正常组织的患者,免疫组化法检测宫颈组织中FHIT蛋白表达;多重PCR法检测HPV16 E2/E7表达。结果:FHIT蛋白的总阳性表达率为57.26%(67/117),正常宫颈组织、CINⅠ、CINⅡ、CINⅢ和宫颈癌中FHIT蛋白阳性率分别为85.00%、80.00%、75.00%、60.00%和26.19%。随着宫颈病变加重,FHIT蛋白阳性表达率下降,组间差异有统计学意义(χ~2=7.335;P=0.003)。117例单纯HPV16阳性标本HPV16总整合率为81.20%,正常宫颈组织、CINⅠ、CINⅡ、CINⅢ和宫颈癌中整合率分别为60.00%、66.67%、75.00%、95.00%和92.86%;随病变加重,整合率增强,组间差异有统计学意义(χ~2=5.713,P=0.003);FHIT蛋白阳性表达在不同HPV16整合时不同,差异有统计学意义(χ~2=11.989,P=0.000)。结论:HPV16基因整合可能通过诱导FHIT基因低表达从而促使宫颈癌发生发展。     

HPV-16 E2结合位点甲基化水平与宫颈病变的相关性研究

目的:探讨HPV-16长调控区(LCR)4个E2蛋白结合位点(E2BSs)甲基化水平与宫颈病变的关系。方法:选取单一型HPV-16感染者的宫颈脱落细胞标本59例,分为3组。焦磷酸测序技术定量检测4个E2BSs序列包含的7个Cp G位点的甲基化水平。b DNA技术检测对应标本内E6/E7 mRNA表达水平。分析E2BSs位点甲基化水平与E6/E7 mRNA水平及宫颈病变程度的相关性。结果:HPV-16 E2BSs甲基化水平与宫颈病变均呈正相关(r_s=0.614,0.599,0.289,0.502,P均0.05),其中E2BS3与宫颈病变间相关性较弱(r_s=0.289,P=0.027)。除E2BS3位点(P=0.070),E2BS 1、2及4位点甲基化水平分别在HPV-16无症状感染组、LSIL组和HSIL+组组间的差异均有统计学意义(P均0.05)。LSIL组中,4个E2BSs位点间的甲基化水平差异有统计学意义(χ~2=17.672,P=0.001),且E2BS1位点甲基化水平高于E2BS 2、3及4位点甲基化水平(P均0.05)。HSIL+组内,4个E2BSs位点间的甲基化水平比较,差异有统计学意义(χ~2=18.387,P=0.000),E2BS1位点甲基化水平高于E2BS3和E2BS4位点(P均0.05),E2BS3位点甲基化水平最低(P均0.05)。E6/E7 mRNA水平随E2BS1和2位点甲基化水平升高而升高(rs=0.480,0.467,P均0.05)。E2BS1和E2BS2位点甲基化水平诊断HSIL和SCC病变的ROC曲线下面积(AUC)较大,AUC分别为0.791(95%CI=0.668~0.914)和0.806(95%CI=0.680~0.931),诊断敏感性均为73.9%,特异性分别为72.2%和80.6%。结论:HPV-16E2BSs基因甲基化状态与宫颈病变程度存在相关性,E2BSs甲基化状态有望作为临床上预测宫颈癌前病变及宫颈癌的早期分子指标。     

青岛地区宫颈病变组织中HPV16 URR和E6基因突变分析

目的:分析青岛地区妇女不同宫颈病变组织中人乳头瘤病毒16型(HPV16)上游调控区(URR)和E6基因序列多态性,以及序列变异与宫颈病变的相关性。方法:采用聚合酶链式反应(PCR)技术检测120例宫颈癌及254例宫颈上皮内瘤变(CIN)标本,筛选出HPV16阳性标本,进而扩增出HPV16URR和E6基因,PCR产物纯化后测序,与德国HPV标准株进行对比分析。结果:URR测序结果发现了19个突变位点,所有标本在7521位点均发生了G→A突变;63.30%(69/109)标本中检测到24,7730和7842的联合突变,为另一突变热点。E6测序结果共发现17个突变位点,突变热点为178位点(D25E),其在宫颈癌组、CINⅢ组和CINⅡ组的比例分别为61.11%(44/72)、62.00%(31/50)和50.00%(16/32),3组的差异无统计学意义(P=0.499)。仅观察到6例350位点突变(L83V)。亚洲型(AS)是最多见的突变类型,在154例HPV16阳性标本中占59.09%,其次为E-P原型(33.12%)。结论:青岛地区妇女宫颈病变组织中HPV16URR突变热点为nt7521以及nt24、nt7730和nt7842的联合突变。HPV16E6突变热点为nt178,这些突变热点可能与癌前病变的进展有密切关系。AS型和原型是青岛地区宫颈病变组织中两种主要的HPV16分支。     

子宫颈癌组织中抑癌基因DNA甲基化的检测

目的 通过分析宫颈上皮内瘤变（CIN）和宫颈癌组织中p16、CDH1、RASSF1A和TIMP3基因DNA甲基化的变化情况,探讨其在宫颈癌发生中的意义。方法 用甲基化特异性聚合酶链反应技术（MSP）对CINI组（40份）、CINⅡ～Ⅲ（40份）、宫颈癌组（40份）病变组织中p16、CDH1、RASSF1A和TIMP3基因DNA甲基化程度进行检测。另取正常宫颈组织20份作为对照组。结果（1）对照组中p16、CDH1、RASSF1A和TIMP3基因的DNA甲基化阳性率均为0。（2）p16和CDH1基因的DNA甲基化阳性率,CIN11～m组（分别为22％、35％）明显高于CINI组（分别为2％、5％,P〈0．05）;RASSF1A和TIMP3基因的DNA甲基化阳性率,CINⅡ～Ⅲ组（分别为12％、15％）虽高于CINI组（均为2％）,但差异无统计学意义（P〉0．05）。（3）宫颈癌组p16（40％）、CDH1（58％）、RASSF1A（20％）和TIMP3（35％）基因的DNA甲基化阳性率虽均高于CINⅡ-Ⅲ组,但差异无统计学意义（P〉0．05）。（4）宫颈癌组p16、CDH1、RASSF1A和TIMP3基因的DNA甲基化阳性率均高于CINI组,差异有统计学意义（P〈0．05）。（5）上述基因的DNA甲基化的总阳性率（即任何一个基因出现甲基化即为阳性）,宫颈癌组（90％）明显高于CINⅡ～Ⅲ组（55％,P〈0．05）,且此两组均明显高于CINI组（8％,P〈0．05）。结论 随着从宫颈不典型增生向浸润癌的进展,p16、CDH1、RASSF1 A和,TIMP3基因的DNA甲基化阳性率明显升高,提示抑癌基因的DNA甲基化在宫颈癌发生、发展中起着一定作用,可能成为判断宫颈癌发生、发展的重要指标。     

TSLC1基因异常甲基化及与HPV的关系在宫颈癌发生发展中的意义

目的:探讨TSLC1基因启动子甲基化状态以及与HPV感染的关系在子宫颈癌发生发展中的意义。方法:用甲基化特异性聚合酶链反应(MSP)法检测68例原发性宫颈癌,67例CIN及29例慢性宫颈炎症石蜡包埋组织中TSLC1基因甲基化状态。用聚合酶链反应(PCR)法检测人乳头瘤病毒(HPV)DNA的感染状况。结果:宫颈癌组织中45例异常甲基化(66.2%),其中鳞癌67.8%(40/59),腺癌55.5%(5/9);67例宫颈上皮内瘤变组织中29例可见异常甲基化(43.3%);29例慢性炎症组织中2例甲基化(6.9%);将鳞癌、腺癌、CIN与慢性炎症比较,鳞癌与CIN比较,CINⅡ～Ⅲ与CINⅠ比较均显示差异有统计学意义(P<0.05),腺癌与CIN比较,鳞癌分化程度之间的差异无统计学意义(P>0.05)。HPV感染率宫颈癌组51.5%(35/68),CINⅠ23.1%(6/26),CINⅡ～Ⅲ41.5%(17/41),慢性炎症组17.2%(5/29);TSLC1甲基化与HPV感染相关性比较,CIN组之间差异有统计学意义(P<0.05),宫颈癌组之间的差异则无统计学意义(P>0.05)。结论:TSLE1基因启动子区CPG岛的高甲基化是导致TSLC1基因失活的重要机制,可能参与宫颈癌的发生;它与HPV之间的关系提示TSLC1基因可能是宫颈细胞癌变过程中的早期事件。     

青岛地区子宫颈病变组织中人乳头瘤病毒16型L1基因多态性分析

目的分析青岛地区不同宫颈病变组织中人乳头瘤病毒（HPV）16型L1基因序列的多态性,探讨其序列变异与宫颈癌的相关性。方法提取2008年6月至2010年5月青岛大学医学院附属医院妇产科61例宫颈癌组织和181例宫颈脱落细胞DNA,采用PCR技术筛选HPV16阳性标本,进而对其L1基因全长进行扩增、测序,应用DNASTAR软件进行序列比对,分析核苷酸和氨基酸的变异。结果慢性宫颈炎组、宫颈上皮内瘤变（CIN）组和宫颈癌组HPV16的感染率分别为30.0%（12/40）、41.1%（58/141）和65.6%（40/61）,3组比较,差异有统计学意义（P〈0.05）。共有96例成功扩增出HPV16L1基因,慢性宫颈炎、CIN和宫颈癌组扩增率分别为25.0%（10/40）、37.6%（53/141）和54.1%（33/61）。HPV16型L1基因共发现13处变异,宫颈癌组织均发生C6240G、A6432G、6902insATC、6954delGAT及G7061A（无义）5处突变,另一突变热点为A6178C,但其在慢性宫颈炎、CIN、宫颈癌组的频率分布比较,差异无统计学意义（P〉0.05）。结论青岛地区HPV16 L1基因的多态性具有地域性,可能导致病毒地方株的免疫原性和抗原性改变,但与宫颈癌的发生发展无明显关系。     

p16基因与宫颈癌和宫颈上皮内瘤样病变诊断

p16基因是一种多肿瘤抑制基因,与多种肿瘤的发生密切相关.p16基因失活导致肿瘤的发生,其失活机制主要为甲基化、缺失及突变.宫颈癌是妇女常见肿瘤之一,发生机制尚不清楚.p16与宫颈癌及宫颈上皮内瘤样病变(CIN)诊断相关性的研究中发现,p16基因的失活及P16的表达很复杂,p16基因在宫颈癌发生、发展中的作用还不清楚.有研究发现p16基因在宫颈癌发展中主要的失活形式是启动子甲基化,失活率随病情恶化而增加;而在人乳头状瘤病毒(HPV)感染的宫颈癌和CIN中,p16基因的缺失和突变是罕见事件,表现为过表达,几乎所有HPV阳性宫颈癌中p16表达阳性.为此对p16基因与宫颈癌、CIN的研究进展进行综述.     

Diagnosis and management of Candida of the nipple and breast

Diagnosis and treatment of ductal and/or nipple candidiasis in breastfeeding women is complicated by the variety of symptoms women experience. The differential diagnosis includes candidiasis of the nipple, candidiasis of the breast, bacterial infection of either nipple or breast, and other less common problems such as Raynaud's syndrome. Diagnosis and treatment are based on history, physical examination, and presenting symptomatology because cultures of breast milk are often inconclusive. Differential diagnoses and treatment options are reviewed.     

Comparison of indications of pregnancy termination and prognosis of mothers and neonates in early- and late-onset preeclampsia

Objective: To compare the indications of pregnancy termination and prognosis between early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP). Methods: In total, 100 patients diagnosed early-onset preeclampsia in our hospital from January 1, 2012, to June 30, 2014, were recruited for this retrospective cohort study. At the same time, we randomly chose another 100 late-onset preeclampsia as the contrast group. Criterion distinguishing early versus late was set at week 34 of gestation. Indications for pregnancy termination and prognosis of mothers and neonates were compared between the groups. Results: Significant differences were observed between the groups regarding indications for terminating pregnancy. The EOP indications to terminate the pregnancy were mainly fetal-related, while LOP were mainly maternal-related. Postpartum neonatal morbidity and mortality were significantly higher, mean gestational age onset and delivery were significantly earlier, latent period for delivery and postpartum hospitalization time were significantly longer, admission 24 h proteinuria was significantly higher in EOP than in LOP group (P < 0.05). Conclusion: EOP is a distinct and more severe clinical entity with earlier gestational age onset and delivery. EOP might be a fetal-related disease complicated by severe placental and perinatal injuries; LOP might be a maternal-related derived disease condition.     

Pharmacokinetics and endometrial and tubal tissue penetration of cefalotin and cefazolin

The pharmacokinetics and concentrations of the two antibiotics cefazolin and cefalotin were studied during gynecologic operations in endometrial and tubal tissue. The patients received 0.05 g/kg of the antibiotics by intravenous injection. Under the given conditions, pharmacokinetic calculation of the plasma elimination gave half-lives of 24.8 min for cefalotin and of 63 min for cefazolin. Fitting of the tissue levels to the Bateman function showed that the two antibiotics diffuse rapidly into both tubal and endometrial tissue and attain peak concentration levels between 10 and 25 min. In both tissues the concentrations of cefazolin were higher than those of cefalotin. Higher tissue concentrations of cefazolin could also be demonstrated in experiments of longer duration.     

Vitamin and homocysteine status of mothers and infants and the risk of nonsyndromic orofacial clefts

OBJECTIVE: This study was undertaken to investigate the involvement of maternal and infant B vitamins and homocysteine as risk factors for orofacial clefting. STUDY DESIGN: Venous blood samples were taken from 96 infants with nonsyndromic orofacial clefts and 88 infants without a congenital malformation and from their mothers at approximately 14 months after the index pregnancy. Red blood cell and serum folate, serum vitamin B(12), whole blood vitamin B(6) as pyridoxal-5'-phosphate (PLP), and plasma homocysteine concentrations were measured. RESULTS: A vitamin B(12) concentration of 185 pmol/L or less and a PLP concentration of 44 nmol/L or less in mothers increased the risk of having a child with an orofacial cleft (odds ratio [OR]=3.1; 95% CI: 1.3-7.4, OR=2.9; 95% CI: 1.2-7.1, respectively). Infants with orofacial clefts had a 15% lower serum folate concentration compared with controls (P=.06). CONCLUSION: A low vitamin B(12) and PLP concentration in mothers increased the risk of orofacial clefts in the offspring. A possible role of the infant's folate status is suggested.     

关于妇产科综合征的理解和应用

综合征(syndrome)来源于希腊语syn("一起"、"共同"之意)和drómos(事件),并起意为"同时发生或同时发生的事件"(concurrence).其实,这也是医学词汇里最古老的名词之一,可以追溯到希腊的波克拉底和盖伦时代,就已被用于描述同时发生的一组症状[1].     

Irisin与糖脂代谢性疾病的研究进展

Irisin是一种新发现的肌肉因子,其前体物质为Ⅲ型纤连蛋白结构域5(FNDC5)。运动使骨骼肌中的过氧化物酶体增殖活化受体γ(PPARγ)辅助激活因子1α(PCG-1α)的表达增加,PCG-1α能诱导FNDC5表达,FNDC5断裂后可形成分泌性的肌肉因子Irisin,从而诱导白色脂肪棕色化,导致能量消耗增多,体质量减轻,改善糖耐量和胰岛素抵抗,调节糖脂代谢和能量代谢。因此,Irisin有望成为治疗糖脂代谢性疾病的新靶点。本文对Irisin与糖脂代谢性疾病的相关性研究进行综述。     

Accuracy and correlates of maternal recall of birthweight and gestational age

Objective  To determine the accuracy of maternal recall of children birthweight (BW) and gestational age (GA), using the Danish Medical Birth Register (DBR) as reference and to examine the reliability of recalled BW and its potential correlates.
Design  Comparison of data from the DBR and the European Youth Heart Study (EYHS).
Setting  Schools in Odense, Denmark.
Population  A total of 1271 and 678 mothers of school children participated with information in the accuracy studies of BW and GA, respectively. The reliability sample of BW was composed of 359 women.
Method  The agreement between the two sources was evaluated by mean differences (MD), intraclass correlation coefficient (ICC) and Bland–Altman's plots. The misclassification of the various BW and GA categories were also estimated.
Main outcome measures  Differences between recalled and registered BW and GA.
Results  There was high agreement between recalled and registered BW (MD =−0.2 g; ICC = 0.94) and GA (MD = 0.3 weeks; ICC = 0.76). Only 1.6% of BW would have been misclassified into low, normal or high BW and 16.5% of GA would have been misclassified into preterm, term or post-term based on maternal recall. The logistic regression revealed that the most important variables in the discordance between recalled and registered BW were ethnicity and parity. Maternal recall of BW was highly reliable (MD =−5.5 g; ICC = 0.93), and reliability remained high across subgroups.
Conclusion  Maternal recall of BW and GA seems to be sufficiently accurate for clinical and epidemiological use.     

PRL-3和MMP-9与子宫内膜异位症侵袭、种植关系的研究

目的:观察促肝细胞再生磷酸酶3(PRL-3)和基质金属蛋白酶9(MMP-9)在子宫内膜异位症(EMs)的表达,探讨它与子宫内膜异位症侵袭、种植的关系。方法:用SABC免疫组织化学法检测PRL-3和MMP-9在31例EMs的异位内膜及在位内膜组织、27例正常子宫内膜组织的表达。结果:PRL-3与MMP-9表达的强弱顺序均为:EMs组异位内膜>EMs组在位内膜>对照组子宫内膜,组间差异有统计学意义(P<0.05);Ⅰ~Ⅱ期的异位内膜PRL-3及MMP-9表达强度均高于Ⅲ~Ⅳ期(P<0.05),但在位内膜二者表达强度在不同临床分期中无统计学差异(P>0.05)。EMs组异位内膜的PRL-3与MMP-9表达呈正相关(P<0.05),在位内膜的PRL-3与MMP-9表达则无明显相关性(P>0.05)。结论:PRL-3和MMP-9可能与EMs的侵袭、种植密切相关。