Inhibiting PI3K/Akt pathway increases DNA damage of cervical carcinoma HeLa cells by drug radiosensitization

This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells.The 50% inhibition concentration(IC50) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay(MTT) in vitro.HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC20 alone or combined with LY294002 for 24 h,and then radiated by different doses of X-ray.The cell survival ratio was obtained by means of clone formation.One-hit multi-target mod...     

P13K/Akt通路在药物放射增敏中作用的机制

目的 进一步探讨PI3K/Akt信号转导途径在药物放射增敏中作用的分子机制.方法 体外培养HeLa细胞,多西紫杉醇(docetaxel)和顺铂(cisplatin)单独及分别联合P13K抑制剂LY294002作用24 h,X线6 Gy剂量照射;Western blot检测Akt、磷酸化Akt(pAkt)、Bad、磷酸化Bad(pBad)蛋白的表达变化;RT-PCR检测Bad、Ku70、Ku80 mR-NA的表达变化;中性彗星电泳检测不同处理组细胞DNA的损伤.结果 ①docetaxel+LY294002联合照射组、cispla-tin+LY294002联合照射组Bad mRNA表达增高,Ku70 mRNA表达减低,Ku80 mRNA表达无明显变化;②docetaxel+LY294002联合照射组、cisplatin+LY294002联合照射组Bad蛋白表达增高,pAkt、pBad蛋白表达减低,Akt蛋白表达无明显变化;③docetaxel+LY294002联合照射组、cisplatin+LY294002联合照射组细胞彗星电泳尾距明显长于单纯药物增敏照射组.结论 ①docetaxel和cisplatin药物增敏照射能够明显活化PI3K/Akt信号转导途径;②抑制PI3K/Akt信号转导途径能够抑制Ku70表达,减少细胞DNA损伤后的再修复,提高促凋亡因子Bad的表达,促进细胞的凋亡.     

The Association between Polymorphism of TNF-α Gene and Hypertensive Disorder Complicating Pregnancy

To study whether the development of hypertensive disorder complicating pregnancy is associated with -308G→A, -850C→T mutation at promoter of TNF-α gene, the -308G→A, -850C→T polymorphism was examined in patients and healthy pregnant women by PCR-RFLP technique. The frequencies of genotype and allele were compared between the two groups. The re- sults showed that with -308G→A polymorphism distribution, the allele frequency of TNF2 and the frequency of the genotype TNF2/1 in the patient group was significantly higher in the patient group than in control group (P<0.05). A significant difference in genotype distribution of -850C→T poly- morphism was observed between the two groups. The allele frequencies of T in patient group was higher in the control group as compared with the patient group. The frequencies of CT and TT genotypes were lower in the patient group. It is concluded that the TNF2 allele of -308 is associated with the occurrence of hypertensive disorder complicating pregnancy, while T allele of -850 may be the protective factor against the development of the disease. TNF2/1 CC may be susceptibility genotype of hypertensive disorder complicating pregnancy.     

母体血浆中游离胎儿DNA在无创性产前诊断中的应用研究

目的：建立一种从孕妇血浆中检测胎儿DNA的无创性产前诊断新方法。方法：对30例孕7－41周妇女血浆游离DNA进行聚合酶链反应，特异性扩增的胎儿基因为Y染色体上的DYZ1位点，扩增片段的大小为149bp.30例孕妇血浆均经过酚氯仿法提取DNA作为模板，进行聚合酶链反应。结果：20例妊娠男性胎儿孕妇中有17例出现DYZ1基因扩增条带，检出率为85．0％，10例妊娠女性胎儿孕妇中未出现阳性扩增条带，无假阳性结果，本研究中早，中，晚孕期性别符合率分别为80％，80％，90％，总符合率为85．0％，结论：用酚氯仿法提取血浆中的DNA，可以提高模板的浓度和纯度，可改善PCR条件，提高结果的准确性。     

瘦素对早孕绒毛滋养细胞分泌CXCR4及MMP9的影响

目的 研究瘦素对早孕绒毛滋养细胞分泌趋化因子受体4(CXCR4)及金属基质蛋白酶-9 (MMP9) 的调节作用.方法 分离、培养早孕期绒毛滋养细胞,添加不同浓度的瘦素,培养24h后,RT-PCR法检测滋养细胞中CXCR4及MMP9 mRNA表达水平.结果 添加了瘦素的滋养细胞表达CXCR4及MMP9 mRNA的量较空白对照组极显著增加(P＜0.01).结论 瘦素能增强滋养细胞分泌CXCR4及MMP9的能力.瘦素可能在孕早期参与了胎盘形成等病理生理过程.     

PI3KAktCOX-2途径在人宫颈癌HeLa细胞放射抗拒中的作用

目的 探讨PI3K/Akt/COX-2途径在人宫颈癌HeLa细胞放射抗拒中作用的分子机制。 方法 COX-2抑制剂塞来昔布单独及联合PI3K抑制剂LY294002作用HeLa细胞24 h,X射线不同剂量照射,利用克隆形成法计算细胞存活率,单击多靶模型拟合细胞存活曲线,计算D_q、D₀、SF₂值和放射增敏比(SER);应用Western blot方法检测Akt、磷酸化Akt、COX-2、Bad和磷酸化Bad蛋白的表达;应用RT-PCR检测COX-2和Bad mRNA的表达。结果 塞来昔布单独及联合LY294002作用HeLa细胞组D_q、D₀、SF₂明显低于单纯照射组;X射线照射后,能够活化HeLa细胞PI3K/Akt/COX-2途径,塞来昔布能够多靶点抑制PI3K/Akt/COX-2途径的活化。结论 PI3K/Akt/COX-2途径的活化是HeLa细胞放射抗拒的重要原因,PI3K/Akt与COX-2之间存在正反馈的调节,抑制PI3K/Akt/COX-2途径的活化能够提高HeLa细胞放射敏感性。     

内皮型一氧化氮合酶基因多态性与妊娠期高血压疾病的相关性研究

目的：探讨内皮型一氧化氮合酶基因(eNOS)第4内含子27bp可变数目的短片段重复序列(variable number tandem repeats,VNTR)的多态性现象与妊娠期高血压疾病(hypertensive disorder complicating pregnancy)的相关性研究,并探讨该位点的多态性在发病机制中的作用。方法：应用聚合酶链反应结合聚丙烯酰胺电泳,银染技术检测患者和健康孕妇的eNOS基因第4内含子多态性。采用RT-PCR技术检测对照组、病例组胎盘中eNOS mRNA的含量,对两组中不同基因型胎盘eNOS mRNA表达水平分别进行半定量比较。结果：eNOS基因VNTR 4a等位基因频率在正常孕妇组为7．50％,明显低于病例组的16．00％(χ^2=4．63,P=0．031,OR值为2．35,95％可信限为4．74～23．2),4a／4b+4a／4a基因型频率在两组分别为28．00％和13．75％(χ^2=5．08,P=0．024,OR=2.69,95％可信限为6.12～35.4)差异具有统计学意义。病例组eNOS mRNA的表达明显低于正常组,病例组中464b基因型组的eNOS mRNA水平明显高于4a／4b+4a／4a基因型组(76．81±1．41vs62．79±0．95,t=11．63 P=0．012)。结论：eNOS基因第4内含子VNTR与妊娠期高血压疾病具有相关性,这可能与该位点的突变可能引起eNOS mRNA转录减少有关。     

无症状沙眼衣原体和解脲支原体感染对早孕绒毛滋养细胞表达CXCL12/CXCR4的影响

目的研究无症状沙眼衣原体和解脲支原体感染患者趋化性细胞因子CXCL12／CXCR4在母胎界面的表达情况，探讨其对早孕绒毛滋养细胞侵蚀力的影响。方法采用免疫组化法观察正常早孕和无症状沙眼衣原体、解脲支原体感染患者早孕绒毛组织中的CXCL12／CXCR4的定位和分布，采用免疫印迹、RT—PCR法检测其蛋白和mRNA的表达情况；将无症状沙眼衣原体和解脲支原体感染患者绒毛外滋养细胞进行分离培养和鉴定，应用免疫细胞化学法检测CXCL12／CXCR4的表达。结果免疫组化检测显示正常妊娠时CXCL12、CXCR4主要表达于绒毛滋养细胞、绒毛间质细胞以及蜕膜组织中；RT—PCR法和Western bolt检测显示：与正常妊娠相比，无症状沙眼衣原体和解脲支原体绒毛感染者的早孕绒毛组织中CXCL12、CXCR4 mRNA和蛋白表达减弱；正常早孕绒毛外细胞滋养细胞中CXCL12、CXCR4均呈强阳性表达，而无症状沙眼衣原体和解脲支原体绒毛感染者其绒毛外细胞滋养细胞中CXCL12、CXCR4表达减弱。结论CXCL12／CXCR4受体配体系统参与了绒毛外滋养细胞的侵蚀，无症状沙眼衣原体和解脲支原体感染状态下可能影响早孕绒毛CXCL12／CXCR4受体配体系统表达，导致母胎界面出现免疫紊乱现象，从而对早孕绒毛滋养细胞侵蚀力造成影响。     

CXCL12/CXCR4 Expression in Trophoblasts Takes Part in Materno-fetal Immune Tolerance and Vascular Remodeling

Immunological tolerance is necessary for materno-fetal interface in pregnancy. The enrichment of CD56brightCD16-NK cells in decidual plays a crucial part in the immunological tolerance[1], which was reported recently to be related with chemokine CXCL12[2]. On the other hand, EVTs exert significant effect on placenta embed during gestation. It is known that CXCL12 is concerned with tumor invasion and metastasis[3], and EVTs have the invasive and migratory ability similar to malignant tu…     

宫颈机能不全紧急环扎术的效果评估——附20例临床分析

目的：探讨宫颈紧急环扎术治疗宫颈机能不全（cervical incompetence,CI）的效果。方法：将我院2005-06-2009-12收治妊娠期宫颈机能不全患者20例分为：选择性环扎组7例,紧急环扎组7例,另外6例纳入保守治疗组作为对照（简称对照组）。入院后完善相关检查,全部手术组实施宫颈环扎术及药物保胎治疗,对照组仅给予保胎药物治疗。上述处理后观察5～7d,无流产征兆者出院,规律产检,随访至妊娠终止。观察其平均保胎天数、新生儿出生体重、新生儿存活、〉34周分娩、〉37周分娩、胎膜早破发生情况,对结果进行统计分析。结果：保胎天数：选择性环扎组（154.86±10.51）d,紧急环扎组（34.86±46.84）d,对照组（94.75±30.93）d;新生儿平均出生体重：选择性环扎组（3171.43±424.12）g,紧急环扎组（1557.14±907.11）g,对照组（2812.50±383.79）g;新生儿存活：选择性环扎组7例,紧急环扎组2例,对照组6例;〉34周者分娩：选择性环扎组7例,紧急环扎组2例,对照组6例;〉37周分娩：选择性环扎组4例,紧急环扎组1例,对照组3例;胎膜早破的发生：选择性环扎组0例,紧急环扎组4例,对照组0例。结论：宫颈机能不全是导致妊娠晚期流产和早产的常见原因,预防性宫颈环扎可延长保胎天数、增加〉34周分娩、减少胎膜早破的发生、增加新生儿平均出生体重,但并不能减少早产的发生。对CI的低危患者,保守治疗也能取得较好的妊娠结局和新生儿结局。