脂肪组织提取细胞及其临床展望

新近发现的脂肪组织提取细胞(PLA细胞)已成为干细胞研究的又一热点。PLA细胞不但具有自体来源、容易获取、再生能力强、低衰老性等特点,而且尚具备多向分化潜能,可定向分化为脂肪细胞、肌细胞、软骨细胞及成骨细胞等。该文对PLA细胞的基本特性及多向分化潜能进行综述,并探讨其在基因治疗中的优势和临床应用前景。     

长效和短效GnRH-a在IVF/ICSI周期长方案中的应用比较

目的:探讨长效和短效促性腺激素释放激素激动剂(GnRH-a,曲普瑞林)在体外授精/卵细胞浆内单精子注射(IVF/ICSI)周期长方案中的应用及其对临床结局的影响。方法:对排卵正常者采用黄体中期降调长方案,无排卵者行口服避孕药(OC)预处理长方案,根据采用不同的剂型分为长效GnRH-a组和短效GnRH-a组,对两组在应用过程中的检测值进行比较。结果:无论黄体中期降调长方案,还是OC预处理长方案,其长效GnRH-a组的Gn用量和Gn天数,注射绒促性素(HCG)日血孕酮水平均明显高于短效GnRH-a组(P<0.05,P<0.01),长效GnRH-a组妊娠率和种植率明显低于短效GnRH-a组(P<0.05)。两种方案中,长效和短效GnRH-a组间按主导卵泡平均直径范围(≥18mm～≤20mm、>20mm～≤22mm和>22mm)比较,各直径范围中周期所占比例差异无统计学意义(P>0.05)。结论:长方案中使用长效GnRH-a降调比短效会增加Gn用量,使孕酮水平升高,妊娠率和种植率降低,对临床结局有不利影响。     

短时受精周期中患者年龄与多原核合子发生及妊娠结局的关系

目的探讨短时受精周期中患者年龄与多原核合子发生及妊娠结局的关系。方法回顾性分析本中心2008年1月至2010年12月共3862例IVF短时受精治疗周期资料,按患者年龄分为<35岁,35-37岁,38-40岁,>40岁四组,分别比较四组患者的多原核合子发生率、妊娠率、种植率、早期胚胎丢失率。结果短时受精患者年龄高于37岁时,其产生多原核合子的周期比例显著降低(P<0.05)。随着患者年龄的增加,妊娠率、种植率均显著性降低(P<0.01),而早期胚胎丢失率显著性升高(P<0.01)。结论短时受精周期中,年龄37岁以上患者周期产生多原核合子概率低于37岁及以下患者,患者的年龄可以作为预测短时受精周期临床结局的指标。     

根据卵泡输出率评估IVF中卵巢的反应性及结局

目的:研究在体外受精-胚胎移植(IVF-ET)中,卵泡输出率(follicular output rate,FORT)对卵巢反应性的评估能力及对妊娠结局的预测能力?方法:共研究3 292例控制性超排卵患者,每位患者均以外源性重组卵泡刺激素(recombinant follicle stimulating hormone,rFSH) 225 iu启动?根据FORT值将其分为低?中?高3组?回顾性研究了3组间控制性超排卵数据及实验室指标?结果:窦卵泡数(antral follicle count,AFC)在高FORT组最低,在低FORT组最高?排卵前卵泡数(preovulatory follicle count,PFC)及注射人体绒毛膜促性腺激素(human chorionic gonadotrophin,HCG)日当天雌二醇 (estradiol,E2)在高FORT组中最高,在低FORT组中最低?获卵数?总胚胎数?优质胚胎数在低?中?高3组间逐步升高,差异有统计学意义?而胚胎种植率及临床妊娠率在3组间亦有统计学差异,在高FORT组中最高?结论:FORT是研究卵巢反应性的有力工具,高FORT组将具有更好的妊娠结局?     

中药复方对戊四氮致痫大鼠海马mGLuR1表达的影响

目的：观察戊四氮（PTZ）致痫大鼠海马神经元代谢型谷氨酸受体（mGluR1）表达以及中药复方AAP的脑保护作用。方法：动物随机分为6组,复制戊四氮致痫大鼠模型;于致痫后12h、2d、5d、7d相应时间点取材,制备脑标本;免疫组化技术检测大鼠海马神经元mGluR1表达。结果：与正常组比较,模型组mGluR1免疫反应阳性表达增加,差异显著（P〈0.05）;与模型组及丙戊酸钠组比较,中药复方AAPl、AAPm、AAPs组海马CA3区mGluR1阳性表达水平降低,差异显著（P〈0.05）。结论：戊四氮致痫大鼠海马mGluR1表达增加,mGluR1可能在PTZ致大鼠癫痫发作中起作用;中药复方AAP可降低mGluR1表达,对癫痫大鼠有脑保护作用。     

Assessment of the clinical effect of Chinese medicine therapy combined with psychological intervention for treatment of patients of peri-menopausal syndrome complicated with hyperlipidemia

<正>Objective:To observe the effect of Chinese medicine therapy combined with psychological intervention(combined therapy) on the clinical symptoms and levels of blood lipids and sex hormones of patients of peri-menopausal syndrome complicated with hyperlipidemia.Methods:With the use of a randomizing digital table method,185 patients that fit the registration standard were randomly assigned to three groups.The 59 cases in Group A were treated with two Chinese patents,Kunbao Pill(坤宝丸) and Modified Xiaoyao Pill(加味逍遥丸);the 63 in Group B received psychological intervention alone;and the 63 in Group C were treated with both(the combined therapy),with the treatment course for all six months.The items of observation included: (1) scoring by SCL-90 on eight factors and seven symptoms;(2) scoring on Chinese medicine symptoms by Kupermann scale,including anxiety and bad temper,scorching sense ation with sweating,dizziness,tinnitus, soreness and weakness of the loin and knees,palpitation,insomnia,lassitude,weakness,and hyposexuality; (3) blood contents of total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C),apoprotein Al(ApoAl) and B(ApoB);(4) levels of sex hormones, including estradiol(E_2),progesterone(P),pituitary prolactin(PRL),follicular stimulating hormone(FSH),and luteinzing hormone(LH) in some randomly selected patients;(5) adverse reaction;and(6) one-year follow-up study on long-term effect.Results:A total of 21 patients(6,8,and 7 cases in Groups A,B,and C,respectively) dropped out;the drop-out rate was insignificant among groups.(1) The markedly effective rates in Group A, B,and C were 26.42%(14/53),18.18%(10/55),and 53.57%(30/56),respectively,and the total effective rates in them were 64.15%(34/53),50.91%(28/55),and 87.50%(49/56),respectively,suggesting the therapeutic efficacy in Group C was significantly better than that in Groups A and B(P0.01).(2) SCL-90 scoring showed that the total scores decreased significantly after treatment in Group C(P0.01),but remained unchanged in Groups A and B(P0.05).(3) Scoring on Chinese medicine symptoms showed the same results as shown by SCL-90 scoring in terms of total scores and individual symptoms,except that menstrual disorder and amenorrhea were unchanged in all three groups(P0.05).(4) Levels of HDL-C,ApoAl,and E_2 increased and those of TG,TC,LDL-C,ApoB,FSH,and LH decreased after treatment in Group C,reaching near normal levels;similar trends of blood lipids were shown in Group A,but the level of sex hormones was unchanged.In Group B all the above-mentioned indices were unchanged(P0.05).(5) A one-year follow-up study showed the markedly effective rate and the total effective rate in Group C were higher than those in the other two groups respectively(P0.01).(6) No adverse reaction was found.Conclusion:Chinese medicine therapy combined with psychological intervention could not only improve the nervous symptoms,but also regulate the blood levels of lipids and sex hormones in patients of peri-menopausal syndrome complicated with hyperlipidemia.     

miR-181a促进人子宫内膜间质细胞蜕膜化催乳素的表达

目的目前虽已发现一些在胚胎着床过程中起重要作用的分子,但微小RNA(microRNA,miRNA)在蜕膜化过程中作用的研究报道较少,文中调查孕鼠围着床期子宫组织中miR-181a的表达规律,并探讨miR-181a对人子宫内膜间质细胞(human endometrial stromal cell,hESC)体外诱导蜕膜化过程中蜕膜化标志基因蜕膜化催乳素(decidual prolactin,dPRL)表达的调控作用。方法收集怀孕0～6.5 d孕鼠子宫组织,采用TRIZOL法提取总RNA,通过实时定量PCR检测子宫组织中miR-181a的表达。制备Ad-miR-181a重组腺病毒,并感染hESC,24 h后采用0.5 mmol/L 8-Br-cAMP和1μmol/L Medroxyproges-terone(MPA)联合进行体外蜕膜化诱导培养;通过化学发光免疫法和实时定量PCR分别检测细胞培养液上清中dPRL的浓度与间质细胞中催乳素(prolactin,PRL)mRNA的表达;同时采用荧光素酶报告基因检测miR-181a对hESC中dPRL(-332/+65)启动子的调控作用。结果早孕小鼠子宫组织中miR-181a的表达高于非妊娠小鼠子宫,且随着妊娠天数的增加呈逐渐上升的趋势,在小鼠着床"窗口期"即孕4.5 d达到高峰(2.60±0.15倍,P<0.01,n=5),孕5.5 d子宫miR-181a的表达开始平稳下降;成功获得滴度为5.19×1010ifu/ml的miR-181a腺病毒,该腺病毒介导的miR-181a过表达显著增加hESC蜕膜化标志基因dPRL mRNA表达水平,增高超过8倍(P<0.01);在8-Br-cAMP和MPA联合诱导hESC体外蜕膜化时,高表达miR-181a可明显增加8-Br-cAMP联合MPA诱导的PRL mRNA表达水平与PRL蛋白分泌(P<0.01)。荧光素酶报告基因进一步证实miR-181a可以激活dPRL(-332/+65)启动子的活性达到2倍(P<0.05)。结论 miR-181a参与8-Br-cAMP和MPA诱导子宫内膜间质细胞蜕膜化过程中PRL的表达调控。     

人类肥胖相关新基因LYRM1真核表达载体的构建及稳定转染3T3-L1细胞系的建立

目的:构建人LYRM1基因的真核表达载体,转染3T3-L1前体脂肪细胞,建立稳定过表达人LYRM1基因的3T3-L1前体脂肪细胞系.方法:运用RT-PCR技术从人网膜脂肪组织中分离LYRM1基因的完整编码框,将其亚克隆到真核表达载体pcDNATM3.1/myc-His B,脂质体转染3T3-L1前体脂肪细胞,通过G418筛选,建立稳定转染的3T3-L1前体脂肪细胞系,并利用Western blot方法鉴定其表达.结果:PCR、酶切鉴定及测序结果表明重组质粒构建正确.建立了稳定转染LYRM1的3T3-L1前体脂肪细胞,成功地表达目的基因.结论:LYRM1真核表达载体的成功构建及稳定转染3T3-L1细胞系的建立为进一步研究其功能奠定了良好的实验基础.     

鲜胚移植后累计妊娠率及其影响因素

目的：探讨体外受精‐胚胎移植妊娠结局及其影响因素。方法回顾性分析1660例鲜胚移植患者临床结局。根据是否临床妊娠分为妊娠组（A组）和非妊娠组（B组）；根据冻胚移植结果，新鲜周期未获得活胎分娩的妇女分为妊娠组（C组）和未妊娠组（D组）。分析影响妊娠结局的相关因素。结果鲜胚移植临床妊娠率60.12％（998／1660），高于冻胚移植临床妊娠率53.15％（253／476）（ P＜0．05）。A组取卵个数、有效胚胎数、有效胚胎比例、内膜厚度及胚胎评分均高于 B组（P＜0．05）。C组有效胚胎数、有效胚胎比例、移植胚胎数均高于D组（P＜0．05）。A组临床妊娠率高于C组（P＜0．05）。第3次冻胚移植时临床妊娠率较第1、2次下降（P＜0．05）。结论应争取鲜胚移植；但鲜胚移植失败患者仍有较高的冻胚移植成功率。2次冻胚移植失败后应考虑特殊原因引起的不孕，必要时重新取卵。