SIRT3在早发型子痫前期胎盘组织中的表达及作用

目的:检测早发型子痫前期(eoPE)患者胎盘组织中SIRT3表达,探讨SIRT3对滋养细胞功能的调控及其可能机制。方法:免疫组化法和免疫印迹法检测eoPE患者胎盘中SIRT3表达。试剂盒检测eoPE患者胎盘组织氧化应激因子水平。应用瞬时转染质粒使HTR-8/SVneo细胞过表达SIRT3,检测滋养细胞的侵袭和迁移能力。通过CoCl₂构建滋养细胞氧化应激模型,研究SIRT3对HTR-8/SVneo细胞抗氧化应激能力的调控。免疫印迹法检测Akt信号通路关键蛋白表达。结果:eoPE患者胎盘组织中SIRT3表达较对照组显著降低,SOD和GSH-Px水平明显降低,MDA水平明显升高。过表达SIRT3可促进HTR-8/SVneo细胞侵袭和迁移,显著缓解CoCl₂诱导的滋养细胞氧化应激。过表达SIRT3可显著上调Akt信号通路上的关键蛋白表达。结论:胎盘组织中SIRT3下调参与eoPE病理过程。SIRT3可能通过Akt信号通路促进滋养层细胞侵袭、迁移以及维持细胞氧化应激处于平衡状态。     

长链非编码RNA中OIP5-AS1在重度子痫前期孕妇胎盘组织中的表达及意义

目的 探讨长链非编码RNA Opa相互作用蛋白5-反义转录物1(Opa-interactingprotein 5 antisense RNA 1, OIP5-AS1)在重度子痫前期孕妇胎盘组织中的表达及其可能作用。方法 采用实时定量荧光（RT-qPCR）法测定30例重度子痫前期孕妇和30例正常胎盘中OIP5-AS1和miR-29b表达情况;采用过划痕实验和Transwell侵袭实验检测缺氧条件下向人绒毛膜滋养层细胞系（HTR-8/SVneo细胞）内转染OIP5-AS1对HTR-8/SVneo细胞迁移和侵袭能力的影响;采用RT-qPCR法测定转染OIP5-AS1和OIP5-AS1基因的小干扰RNA(OIP5-AS1siRNA)后HTR-8/SVneo细胞微小RNA(microRNA,miRNA)miR-29b的表达情况。结果 在mRNA水平上,重度子痫前期组胎盘组织中OIP5-AS1较对照组表达降低（P <0.01）;过表达OIP5-AS1后,HTR-8/SVneo细胞迁移和侵袭能力增加,差异有统计学意义（P <0.01）;过表达OIP5-AS1后HTR-8/SVneo细胞中...     

过表达miR-198通过靶向调控Wnt2 B表达抑制滋养细胞HTR-8/SVneo的增殖、转移与侵袭

目的:研究过表达miR-198对人滋养细胞株HTR-8/SVneo增殖、转移和侵袭能力的影响,并探讨其可能作用机制.方法:qRT-PCR检测子痫前期(PE)患者及正常产妇胎盘组织中miR-198表达水平.培养人滋养细胞株HTR-8/SVneo,采用脂质体法将miR-198 mimics及其阴性对照mimics NC转染...     

胎盘生长因子在妊娠期糖尿病患者胎盘中抑制滋养细胞凋亡的作用研究

目的:探讨胎盘生长因子(PLGF)对妊娠期糖尿病(GDM)患者胎盘滋养细胞凋亡的影响。方法:收集20例GDM和20例正常孕妇胎盘组织,采用HE染色法观察GDM患者胎盘组织形态学变化。TUNEL染色以及Caspase-3活性检测胎盘中细胞凋亡情况。Western blot检测胎盘中PLGF蛋白表达情况,免疫组化观察PLGF的原位表达。离体培养人绒毛滋养细胞(HTR-8/SVneo),分为对照组、高糖组(HG)、PLGF+高糖组(PLGF+HG)和PLGF组,应用流式细胞术检测各组HTR-8/SVneo滋养细胞的凋亡率。结果:与对照组相比,GDM患者胎盘中滋养细胞凋亡明显减少,Caspase-3活性明显减弱。PLGF在GDM胎盘中的表达明显高于对照组;高糖诱导HTR-8/SVneo滋养细胞凋亡增加,PLGF预处理明显逆转高糖诱导的HTR-8/SVneo滋养细胞凋亡增加。结论:GDM患者胎盘中滋养细胞凋亡明显减少,而胎盘中PLGF表达升高。离体实验进一步证明PLGF能够抑制高糖所致HTR-8/SVneo滋养细胞凋亡增加,因此PLGF表达升高可能是GDM患者胎盘中滋养细胞凋亡减少的原因。     

PTEN在子痫前期胎盘滋养细胞浸润不足中的作用及机制研究

目的:检测子痫前期(PE)患者胎盘组织中PTEN表达,探讨PTEN对滋养细胞迁移功能的调控及其可能机制。方法:收集重度PE患者和正常孕妇的胎盘组织各20例,实时定量PCR和Western blot法检测胎盘组织中PTEN表达;将PTEN过表达质粒(pcDNA3.1-HA-PTEN)或特异性siRNA(siPTEN)瞬时转染至HTR-8/SVneo细胞,同时转染pcDNA3.1或siCTL作为对照。采用划痕实验评估细胞迁移能力,Western blot法检测p-Akt(Thr308)、p-Akt(Ser473)、Akt、MMP-2和MMP-9表达。结果:重度PE患者胎盘组织中PTEN mRNA和蛋白表达明显高于正常孕妇,差异均有统计学意义(P0.01)。与转染pcDNA3.1组比较,pcDNA3.1-HA-PTEN瞬时转染HTR-8/SVneo细胞48h后,细胞迁移率明显降低[(26.42±6.68)%vs(52.92±6.71)%,P0.01],p-Akt(Thr308)和p-Akt(Ser473)表达显著下调,Akt表达无明显改变,MMP-2和MMP-9蛋白表达下降;与转染siCTL组比较,siPTEN瞬时转染HTR-8/SVneo细胞48h后,细胞迁移率则明显升高[(50.39±6.84)%vs(30.04±5.40)%,P0.01],p-Akt(Thr308)和p-Akt(Ser473)的表达明显上调,Akt表达无明显改变,MMP-2和MMP-9蛋白表达升高。结论:重度PE患者胎盘组织中PTEN表达明显升高,PTEN可通过下调Akt活性降低MMP-2和MMP-9表达,抑制滋养细胞的迁移。提示PTEN在PE的发病中发挥了一定的作用。     

PTEN在子痫前期胎盘滋养细胞浸润不足中的作用及机制研究26

目的:检测子痫前期(PE)患者胎盘组织中PTEN表达,探讨PTEN对滋养细胞迁移功能的调控及其可能机制.方法:收集重度PE患者和正常孕妇的胎盘组织各20例,实时定量PCR和Western blot法检测胎盘组织中PTEN表达;将PTEN过表达质粒(pcDNA3.1-HA-PTEN)或特异性siRNA(siPTEN)瞬时转染至HTR-8/SVneo细胞,同时转染pcDNA3.1或siCTL作为对照.采用划痕实验评估细胞迁移能力,Western blot法检测p-Akt(Thr308)、p-Akt(Ser473)、Akt、MMP-2和MMP-9表达.结果:重度PE患者胎盘组织中PTEN mRNA和蛋白表达明显高于正常孕妇,差异均有统计学意义(P<0.01).与转染pcDNA3.1组比较,pcDNA3.1-HA-PTEN瞬时转染HTR-8/SVneo细胞48 h后,细胞迁移率明显降低[(26.42±6.68)%vs(52.92±6.71)%,P<0.01],p-Akt(Thr308)和p-Akt(Ser473)表达显著下调,Akt表达无明显改变,MMP-2和MMP-9蛋白表达下降;与转染siCTL组比较,siPTEN瞬时转染HTR-8/SVneo细胞48 h后,细胞迁移率则明显升高[(50.39±6.84)%vs(30.04±5.40)%,P<0.01],p-Akt(Thr308)和p-Akt(Ser473)的表达明显上调,Akt表达无明显改变,MMP-2和MMP-9蛋白表达升高.结论:重度PE患者胎盘组织中PTEN表达明显升高,PTEN可通过下调Akt活性降低MMP-2和MMP-9表达,抑制滋养细胞的迁移.提示PTEN在PE的发病中发挥了一定的作用.     

子痫前期患者胎盘组织中松弛素受体的表达

目的 探讨子痫前期患者胎盘组织中松弛素受体LGR7表达与子痫前期发病的关系.方法 应用链霉菌抗生物素蛋白-过氧化物酶连接法和RT-PCR技术检测20例正常孕妇(正常组)及26例重度子痫前期孕妇(子痫前期组)胎盘组织中松弛素受体LGR7蛋白及mRNA的表达.结果 (1)两组孕妇的胎盘组织中均有松弛素受体LGR7表达,主要定位于胎盘绒毛滋养细胞的细胞膜上,在细胞滋养细胞、合体滋养细胞中都有较高的表达水平.(2)正常组胎盘组织中松弛素受体LGR7蛋白的表达水平为0.912±0.003,子痫前期组为0.625±0.037,两组比较,差异有统计学意义(P＜0.01).(3)正常组胎盘组织中松弛素受体LGR7 mRNA表达水平为0.776±0.021,子痫前期组为0.393±0.075,两组比较,差异有统计学意义(P＜0.01).结论 子痫前期患者胎盘组织中松弛素受体表达水平下降,可能与子痫前期的发病有关.     

piR-3127964经PIWIL-3/载唾液酸蛋白途径调控滋养细胞侵袭力的机制

目的研究子痫前期与非病理妊娠胎盘组织之间差异表达的piR-3127964调控滋养细胞侵袭力的分子机制。方法收集2020年11月至2021年8月于重庆医科大学附属第一医院产科就诊的非病理性早产(对照组, n=12)与子痫前期孕妇(子痫前期组, n=10)经剖宫产分娩的胎盘以及早孕期人工流产者的绒毛组织(早孕期人工流产组, n=10)。提取对照组和子痫前期组胎盘组织的总RNA进行测序, 利用实时荧光定量聚合酶链反应(real-time quantitative-polymerase chain reaction, qRT-PCR)检测piR-3127964在各类组织中的相对表达量。利用蛋白质印迹法检测3组中PIWIL-1、PIWIL-2和PIWIL-3的表达量。外源性调节HTR-8/SVneo细胞内piR-3127964的水平, 通过qRT-PCR检测下游候选靶分子, 即鸟嘌呤核苷酸结合蛋白样3-样蛋白(guanine nucleotide-binding protein-like 3-like, GNL3L)和载唾液酸蛋白(sialophorin, SPN)mRNA的相对表达量, 利用q...     

子痫前期患者胎盘组织中Rho激酶Ⅱ表达与滋养细胞凋亡的研究

目的探讨Rho激酶Ⅱ(Rho-associatedProteinKinaseⅡ,ROCKⅡ)在正常妊娠和子痫前期患者胎盘的分布和表达及与滋养细胞凋亡的关系。方法采用免疫组织化学法检测30例子痫前期(轻、重度)孕妇和15例正常妊娠妇女胎盘中ROCKⅡ的分布和表达,采用TUNEL法检测三组胎盘滋养细胞的凋亡情况。结果ROCKⅡ在正常妊娠和子痫前期患者胎盘的滋养细胞、内皮细胞、基质细胞中均有表达,以滋养细胞表达为主。子痫前期重度和轻度组以及正常妊娠组ROCKⅡ的免疫组化积分分别为(4.53±0.58)、(3.18±0.59)和(2.06±0.63),P<0.01。正常妊娠和子痫前期患者胎盘的滋养细胞、内皮细胞、基质细胞均存在凋亡,以滋养细胞凋亡为主。重度子痫前期组胎盘滋养细胞凋亡(0.28±0.03)%显著高于子痫前期轻度组(0.22±0.04)%,子痫前期轻度组显著高于正常妊娠组(0.16±0.05)%(P<0.01)。滋养细胞ROCKⅡ表达和滋养细胞凋亡具有相关性(r=0.96,P<0.01)。结论子痫前期患者胎盘组织中Rho激酶Ⅱ的高表达可能与其滋养细胞凋亡增加有关。     

缺氧通过TrkB抑制人滋养细胞系HTR-8/SVneo细胞增殖和侵袭

目的:探讨缺氧对人滋养细胞系HTR-8/SVneo细胞增殖和侵袭的影响及可能的分子机制。方法:将人滋养细胞系HTR-8/SVneo细胞分为缺氧培养(实验组)和常氧培养(对照组),培养48h后,采用CCK8法检测细胞增殖能力,采用Transwell实验评估细胞侵袭能力;采用RT-PCR和Western blot法分别检测HIF-1αmRNA、p-TrkB、TrkB、p-Akt和Akt表达。结果:缺氧处理显著抑制了HTR-8/SVneo细胞的增殖和侵袭能力(P0.05);48h的缺氧处理降低了HIF-1αmRNA表达;缺氧处理48h后,p-TrkB表达显著减少(P0.05);同时Akt的磷酸化水平明显被抑制(P0.05)。结论:长时间持续缺氧通过抑制HTR-8/SVneo细胞中TrkB的活化,进而抑制PI3K/Akt通路的激活来抑制人滋养细胞的增殖和侵袭。     

Differences in female and male perception of information and decision-making in single-embryo transfer in in vitro fertilization in Sweden

Purpose The aim of this study was to evaluate the information and the factors that contribute to the decision to accept and choose single embryo transfer (SET) in females and males. Materials and methods Fifty-four females and males undergoing SET were interviewed separately using a structured questionnaire. Results The women were significantly more satisfied with the information than the men (odds ratio 3.3), but the decision to accept SET was nevertheless more difficult for women (OR 3.1). Only one-third of both female and males were aware of the increased maternal risks with twin pregnancies. There was a tendency that the women who accepted SET had previous children, shorter duration of infertility, and were younger. Cryopreservation of embryos and a good pregnancy chance were important irrespective of gender. Conclusion The female needs more support to choose SET. The male needs better information and further involvement in decision-making. The females were more aware of the fetal risks, but the awareness of the increased maternal risks with twin pregnancies was low. The female need more support to accept and choose single embryo transfer, compared to the male and information should in some areas be directed differently to females and males.     

Non-persistent pesticides in breast milk in an agricultural area in Turkey

Background: Organophosphates, pythyreoids, carbamate pesticides and fungicides are heavily used in agriculture. They may have dangerous effects on newborn health especially on immune system and growth via prenatal transmission by placenta or postnatal transmission by breastfeeding.

Methods: In 2015, 144 non-persistent pesticides in 64 milk samples of 32 mothers were studied by OuEChERS method in liquid chromatography/tandem mass spectrometer in neonatal Intensive Care Unit in Adana, a city in Cukurova region which is an important agricultural area in Turkey.

Results: Pesticides were detected in milk samples of 11 mothers (34.3%) and 21 (32.8%) of milk samples. In five mothers, fungicides (in 5/10 samples propicanozole-PP, in 4/10 samples bromucanozole-BM), in five mothers, organophosphates (in 10/10 samples primyphosphomethyl-PPM), in one mother, both organophosphates and fungicide (in 1/2 samples PPM and in 1/2 samples buprimate) were detected. However, the estimated daily intakes (EDI) were less than acceptable daily intakes (ADI) for PPM, PP and BM, respectively.

Conclusions: Although pesticides levels in human milk did not exceed the ADIs, we suggest monitoring pesticides in human breast milk especially for newborn health.     

Ovulation induction in in vitro fertilization

The main goal of an in vitro fertilization (IVF) program is to maximize the number of women who become pregnant in any treatment cycle. In order to achieve pregnancy, many steps should be successfully accomplished. The number of mature oocytes that are retrieved and fertilized in a treatment cycle has the major impact on the success rate of the IVF program. The chances of achieving conception increases dramatically when the number of embryos replaced into the uterus increases^1-3. Hence, most IVF treatment programs currently use some combination of ovulation induction agents in order to stimulate and aspirate as many follicles as possible.     

Heterogeneity in alpha-thalassemia interactions in Malays, Chinese and Indians in Malaysia

AIM: Interactions between different determinants of alpha-thalassemia raises considerable problems, particularly during pregnancies where antenatal diagnosis is necessary. This study aims to determine the different types of deletional alpha-thalassemia and Hemoglobin Constant Spring (HbCS), and their frequency in Malays, Chinese and Indians in Malaysia. METHODS: DNA from 650 pregnant women from the Antenatal Clinic of the University of Malaya Medical Center in Kuala Lumpur, Malaysia who showed mean cell volume < or =89 fL and/or mean cell hemoglobin < or =28 pg were analyzed for the double alpha-globin gene South-East Asian deletion (--SEA), the -alpha3.7 and -alpha4.2 single alpha-globin gene deletions and HbCS. RESULTS: One hundred and three (15.8%) of the pregnant women were confirmed as alpha-thalassemia carriers: 25 (3.8%) were alpha-thalassemia-1 carriers with the --SEA/alphaalpha genotype, 64 (9.8%) were heterozygous for the -alpha3.7 rightward deletion (-alpha3.7/alphaalpha), four (0.6%) were heterozygous for the -alpha4.2 leftward deletion (-alpha4.2/alphaalpha), nine (1.4%) were heterozygous for HbCS (alphaCSalpha/alphaalpha) and one (0.2%) was compound heterozygous with the -alpha3.7/alphaCSalpha genotype. The double alpha-globin gene --SEA deletion was significantly higher in the Chinese (15%) compared to the Malays (2.5%) and not detected in the Indians studied. The -alpha3.7 deletion was distributed equally in the three races. HbCS and -alpha4.2 was observed only in the Malays. CONCLUSION: The data obtained gives a better understanding of the interactions of the different alpha-thalassemia determinants in the different ethnic groups, thus enabling more rapid and specific confirmation of alpha-thalassemia in affected pregnancies where antenatal diagnosis is necessary.     

缺氧缺血性脑病的性别差异性研究进展

新生儿缺氧缺血性脑病(hypoxic-ischemic encephalopathy,HIE)是指围产期窒息缺氧导致的脑缺氧缺血性损害,可遗留不同程度的神经系统后遗症。动物研究表明,缺氧缺血性损伤后,炎性反应、氧化应激和细胞死亡途径等关键病理生理过程中存在明显的性别差异,雌性动物对亚低温、促红细胞生成素、脑源性营养因子和别嘌呤醇等治疗效果也明显优于雄性。临床研究发现男性HIE患儿病情更重、预后更差。基于性别的治疗干预很有可能在围产期脑损伤中提供更好的神经保护。本文总结了目前HIE性别差异性的相关证据,以期为临床治疗提供新思路。     

Trends in preterm birth in singleton deliveries in a Hong Kong population

Objective

To examine trends in preterm birth and its relationship with perinatal mortality in Hong Kong.

Methods

In a retrospective cohort study, data were reviewed from singletons delivered between 1995 and 2011 at a university teaching hospital. Trends in preterm birth (between 24 and 36 weeks of pregnancy), perinatal mortality, and subtypes of preterm birth (spontaneous, iatrogenic, and following preterm premature rupture of membranes [PPROM]) were examined via linear regression.

Results

There were 103 364 singleton deliveries, of which 6722 (6.5%) occurred preterm, including 1835 (1.8%) early preterm births (24–33 weeks) and 4887 (4.7%) late preterm births (34–36 weeks). Frequency of preterm birth remained fairly consistent over the study period, but that of spontaneous preterm birth decreased by 25% (β = –0.83; P < 0.001), from 4.5% to 3.8%. Frequency of preterm birth following PPROM increased by 135% (β = 0.82; P < 0.001), from 0.7% to 1.7%. The perinatal mortality rate decreased from 56.7 to 37.0 deaths per 1000 deliveries before 37 weeks (β = –0.16; P = 0.54). Early preterm birth contributed to 16.0% of all deaths.

Conclusion

Although the overall rate of preterm birth in Hong Kong has remained constant, the frequencies of its subtypes have changed. Overall perinatal mortality is gradually decreasing, but early preterm birth remains a major contributor.