Study on apoptosis and expression of modulin Bcl-2, Bax in villus syncytiotrophoblast cells in 30 patients with spontaneous abortion

目的 通过观察绒毛合体滋养细胞凋亡以及凋亡调控蛋白Bcl-2和Bax在自然流产患者中的表达,探讨细胞凋亡在自然流产的发病机制.方法 采用原位末端标记法(TUNEL)和免疫组织化学法对30例自然流产患者绒毛合体滋养细胞中调亡指数及凋亡调控蛋白Bcl-2和Bax阳性表达率进行检测,并以30例人工终止早孕的健康妇女做对照(对照组),所有数据采用SPSS16.0进行统计学分析.结果 自然流产组绒毛滋养细胞中凋亡指数为(32.45±5.87)％,明显高于对照组的(19.38±4.16)％,差异有统计学意义(P＜0.01).Bax和Bcl-2在两组合体滋养细胞中的阳性率比较显示,自然流产组中Bax阳性率增高,Bcl-2阳性率下降,Bcl-2/Bax比值升高,差异均有统计学意义(P＜0.01).结论 绒毛合体滋养细胞凋亡明显增加、Bax阳性率增高和Bcl-2阳性率下降在自然流产中产生重要作用.     

30例自然流产患者绒毛滋养细胞凋亡及调控蛋白Bcl-2、Bax的表达研究

目的通过观察绒毛合体滋养细胞凋亡以及凋亡调控蛋白Bcl-2和Bax在自然流产患者中的表达,探讨细胞凋亡在自然流产的发病机制。方法采用原位末端标记法(TUNEL)和免疫组织化学法对30例自然流产患者绒毛合体滋养细胞中调亡指数及凋亡调控蛋白Bcl-2和Bax阳性表达率进行检测,并以30例人工终止早孕的健康妇女做对照(对照组),所有数据采用SPSS16.0 进行统计学分析。结果自然流产组绒毛滋养细胞中凋亡指数为(32.45±5.87)%,明显高于对照组的(19.38±4.16)%,差异有统计学意义(P<0.01)。Bax和Bcl-2在两组合体滋养细胞中的阳性率比较显示,自然流产组中Bax阳性率增高,Bcl-2阳性率下降,Bcl-2/Bax比值升高,差异均有统计学意义(P<0.01)。结论绒毛合体滋养细胞凋亡明显增加、Bax阳性率增高和Bcl-2阳性率下降在自然流产中产生重要作用。     

人类白细胞抗原G和E在人早孕胎盘组织中的表达及其与反复自然流产的关系

目的 探讨人类白细胞抗原(HLA)-G、E在人早孕胎盘组织中的表达分布及其与反复自然流产(RSA)发生的关系.方法 选择15例正常早孕妇女及15例RSA患者为研究对象.分别采用原位杂交方法及免疫组织化学法检测HLA-G、E的mRNA和蛋白质在早孕胎盘组织中的表达.结果 在人早孕胎盘组织所有的滋养细胞,包括合体滋养细胞、绒毛细胞滋养细胞、绒毛外滋养细胞中均有HLA-G、E mRNA的表达.特异性针对HLA-G的单克隆抗体4H84仅在绒毛外滋养细胞中检测出HLA-G蛋白表达.HLA-E蛋白表达与其mRNA表达部位一致.RSA组-眦A-G、EmRNA的表达强度明显低于正常对照组.HLA-G、E蛋白的表达水平在RSA组也明显降低.结论 HLA-G mRNA与蛋白表达部位的不一致可能与单克隆抗体4H84有关.母胎界面的滋养细胞表面HLA-G、E mRNA及蛋白表达水平的下降与RSA的发生密切相关.     

子痫前期患者胎盘组织中细胞间黏附分子1的表达

目的:检测子痫前期患者和正常孕妇胎盘组织中细胞间黏附分子-1(ICAM-1)的表达水平,探讨其在子痫前期发病机制中的作用.方法:采用免疫组化PV-9000法检测50例子痫前期患者(其中轻度子痫前期20例,重度子痫前期30例)和30例正常孕妇子宫胎盘绒毛合体滋养细胞ICAM-1的表达情况.结果:重度子痫前期患者胎盘绒毛合体滋养细胞ICAM-1的表达明显高于正常孕妇组和轻度子痫前期组(P＜0.05),轻度子痫前期组与正常孕妇组ICAM-1的表达差异无统计学意义(P＞0.05).结论:重度子痫前期患者胎盘绒毛合体滋养细胞ICAM-1的表达增强,可能通过引起母胎界面炎症反应而参与子痫前期的发病.     

MMP-25在早孕绒毛和蜕膜中的表达及意义的研究

目的研究基质金属蛋白酶-25 (matrix metalloproteinase-25，MMP-25)在正常早期妊娠和自然流产患者绒毛及蜕膜组织中的表达特点，探讨其在胚胎植入中的作用及其与自然流产的关系。方法采用半定量RT-PCR和免疫组化法检测60例正常早期妊娠和40例自然流产患者绒毛及蜕膜组织中MMP-25mRNA及蛋白的表达。结果① MMP-25mRNA在正常早期妊娠的绒毛及蜕膜组织中均有表达，且绒毛组织中MMP-25mRNA表达水平随妊娠周数的增加逐渐上升，各孕周组间相比差异有统计学意义(P＜0.05)。自然流产组绒毛及蜕膜组织中MMP-25mRNA的表达明显低于正常妊娠组(P＜0.05)。② 正常妊娠组的绒毛组织中，MMP-25主要定位于细胞滋养细胞和合体滋养细胞，并且随妊娠周数增加表达逐渐增强，与RT-PCR结果吻合；自然流产组仅在合体滋养细胞中有表达，两组平均光密度分别为0.21±0.02和0.13±0.04，差异有统计学意义(P＜0.05)。蜕膜组织中MMP-25主要在腺上皮细胞表达，自然流产组的表达微弱，两组平均光密度分别为0.25±0.13和0.21±0.12，差异有统计学意义(P＜0.05)。结论MMP-25表达变化与滋养细胞浸润能力相吻合，且蜕膜组织中呈阳性表达，推测其参与胚胎植入及胎盘形成过程中滋养细胞浸润活性调控及子宫内膜组织重构；自然流产患者MMP-25表达水平降低，造成滋养细胞浸润能力下降，可能与自然流产的发生有关。     

IVF-ET术后流产患者绒毛和蜕膜组织凋亡及相关调控蛋白的表达

目的:检测体外受精-胚胎移植(IVF-ET)术后早期自然流产患者绒毛和蜕膜组织凋亡及相关调控蛋白血管内皮生长因子(VEGF)、Bcl-2的表达,探讨其发病和基因调控机制。方法:采用免疫组织化学染色技术检测IVF-ET术后早期自然流产者(IVF-ET组)绒毛及蜕膜组织Bcl-2、VEGF的表达水平,并以同期正常早孕者(正常早孕组)的标本为对照。结果:VEGF主要表达于绒毛滋养细胞和血管内皮细胞胞浆。IVF-ET组VEGF表达明显低于正常早孕组,差异具有统计学意义(P<0.05)。Bcl-2主要在绒毛合体滋养细胞上表达。IVF-ET组滋养细胞及蜕膜细胞染色较浅,表达呈弱阳性,正常早孕组染色较深,表达强阳性,差异有统计学意义(P相似文献   

原位RT-PCR方法检测妊高征患者胎盘内皮型一氧化氮合酶-mRNA的表达

目的：检测内皮型一氧化氮合酶-mRNA(eNOSmRNA)在妊高征患者胎盘组织中的定位和表达水平的变化。方法：采用标记引物的直接原位RT—PCR方法检测eNOSmRNA在40例妊高征患者与30例正常足月孕妇胎盘组织的定位和表达，并利用IMAGER—PRO PLUS图像分析处理系统对eNOSmRNA在妊高征与正常足月胎盘合体滋养细胞表达水平进行定量分析。结果：eNOSmRNA主要定位在正常足月孕妇和妊高征患者胎盘绒毛的合体滋养细胞。妊高征患者胎盘绒毛合体滋养细胞eNOSmRNA阳性染色减弱，而干绒毛和终末绒毛血管内皮细胞eNOSmRNA阳性染色增强。定量分析表明，妊高征患者胎盘合体滋养细胞eNOSmRNA表达低于正常足月孕妇。结论：eNOSmRNA主要定位在胎盘绒毛的合体滋养细胞，妊高征患者胎盘绒毛合体滋养细胞eNOSmRNA表达水平的下降可能在妊高征的发病中起重要作用。     

Ki-67的表达在胎盘绒毛滋养细胞生存活性评价中的作用

目的：探讨Ki-67的表达在胎盘绒毛滋养细胞生存活性评价中的作用.方法：应用免疫组化MaxVision法检测Ki-67在48例自然流产、48例葡萄胎和25例早期正常妊娠（作为对照组）妇女的胎盘绒毛滋养细胞中的表达情况.结果：葡萄胎组中Ki-67强阳性表达显著高于对照组和自然流产组（P＜0.01）.结论：在胎盘绒毛滋养细胞中,Ki-67表达降低可能导致自然流产,过表达则可导致肿瘤的发生.     

镉对小鼠睾丸间质细胞睾酮合成的影响

目的 探讨镉对小鼠睾丸间质细胞睾酮合成的影响.方法 镉处理组采用20 μmol/L CdCl2处理TM3细胞,分别在加入CdCl2 4 h和8 h后收集细胞上清液,对照组TM3细胞给予等容积磷酸盐缓冲液(DPBS).采用ELISA法测定细胞上清液中睾酮含量,采用RT-PCR和Western blot技术检测睾酮合成关键酶mRNA和蛋白表达水平.结果 镉处理组细胞上清液中睾酮含量明显降低(P＜0. 01).与对照组相比,镉显著下调TM3细胞类固醇激素合成急性调控蛋白(StAR)、细胞色素P450胆固醇侧链裂解酶(P450scc)与 17β-羟类固醇脱氢酶(17β-HSD) mRNA表达(P＜0.01),镉处理组TM3细胞StAR、P450SCC、细胞色素P45017α-羟化酶(P45017α)、3β-HSD与17β-HSD的蛋白表达水平也明显降低,与对照组相比,差异均有统计学意义(P＜ 0.05,P＜0.01).结论 镉抑制小鼠睾丸间质细胞睾酮的分泌,其原因可能与镉抑制间质细胞部分睾酮合成关键酶mRNA和蛋白表达水平有关.     

ES细胞体外定向分化为睾丸间质样细胞模型的建立

目的:建立小鼠胚胎干(embryonic stem,ES)细胞体外分化为睾丸间质样细胞(leydig-like cells)的模型。方法:采用脂质体转染方法将含类固醇生成因子1(steroidogenic factor 1,SF-1)基因片断的质粒转入小鼠ES-D3细胞,并在RA和8Br-cAMP作用下,体外诱导ES-D3细胞分化为睾丸间质样细胞,以空质粒转染组做阴性对照;通过光镜观察转染SF-1的ES-D3细胞定向分化为睾丸间质样细胞过程中的形态变化;用RT-PCR及Western Blot法检测分化过程中类固醇激素合成急性调节蛋白(StAR),胆固醇侧链裂解酶P450scc(CYP11A1)和3β-羟甾脱氢酶1(3β-HSD1)表达情况;免疫荧光法检测睾丸间质样细胞特异性蛋白P450scc及3β-HSD1表达。结果:ES-D3细胞转染SF-1质粒24 h,即检测到SF-1基因表达。转染成功的ES-D3细胞在RA和8Br-cAMP诱导分化48 h后出现睾丸间质样细胞。分化而来的睾丸间质样细胞胞体大而圆润,长有触角,呈纺锤形;且表达睾丸间质样细胞特异性蛋白P450scc和3β-HSD1,同时StAR表达显著增加,而对照组细胞未表达睾丸间质样细胞特异性标志物StAR、P450scc和3β-HSD1。结论:通过对ES-D3细胞转染SF-1质粒并予以RA和8Br-cAMP诱导,成功建立ES-D3细胞定向分化为睾丸间质样细胞模型。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.