转基因显微注射中的基因装液方法改良

目的 对转基因注射过程中的基因装液方法 进行改良.方法 预先从显微注射针背部装入石蜡油至直管部,再使用可编程微量注射仪从注射针针尖倒吸入基因液.结果 基因液顺利装入显微注射针,后续进行显微注射操作的压力状态稳定,避免溶液回吸.结论 本法简便有效地将注射用基因液装入注射针,确保基因液注入原核,有利于提高转基因注射的成功率.     

人卵母细胞玻璃化冷冻的临床应用及成功分娩

目的探讨人卵母细胞玻璃化冷冻保存的方法及临床应用价值。方法将从促排卵周期获得的人的卵母细胞用冷冻环进行玻璃化冷冻保存,复苏后行卵细胞浆内单精子注射-胚胎移植,观察复苏后卵母细胞形成胚胎的发育能力。结果冷冻后融解235个卵母细胞,存活169个（71．9％,169／235）,受精122个（72．2％,122／169）。移植11例,获得临床妊娠2例。第1例于2005年12月15日妊娠35^＋2周行剖宫产,为双胎女婴,均发育正常。第2例为单胎妊娠,现妊娠16周,目前胎儿及孕妇健康。结论卵母细胞玻璃化冷冻是一种较理想的保存女性生育力的方法,但是其技术应用于临床有待进一步研究和实践。     

Pregnancies after Transfer of Vitrified Biopsied Blastocysts

Objective To show two cases describing the successful pregnancies after transplan- tation of vitrified biopsied embryos after preimplantation genetic diagnosis （PGD）. Methods PGD by day 3 embryo biopsy was performed. Excess embryos were frozen using vitrification method. Four months later, the patient elected to undergo a frozen- thawed embryo transfer （FET） cycle. Results These two patients became pregnant and both of them delivered healthy PGD baby respectively. Conclusion This report shows that vitrification is an efficient and practical method for embryo cryopreservation during PGD. Frozen-thawed blastocysts that do survive obtained from vitrified D3 biopsied embryos are able to implant.     

Mechanism of lead induced effects on human spermatozoa after occupational exposure

Objectives: Occupational lead exposure caused several types of male reproductive impairments in different working populations. In the present study we examined the paint factory workers of active reproductive age and compared the data with the non-occupationally exposed desk job holders taken as control from Bangalore, India. Materials and methods: In the above perspective, sperm cell morphology, morphometery and motile activity were assessed. Routine seminal biochemistry, cell cycle phase analysis of sperm head DNA, estimation of serum reproductive hormones and metal levels in blood and semen were also taken into account. Result: Low sperm velocity, ATPase activity, gross and forward progressive motility with high stationary motile spermatozoa revealed lowering of cellular activity after lead exposure (p<0.001), which was supported by high seminal plasma fructose level (p<0.001). Lowering of seminal plasma total protein with concomitant rise in free amino acid level was prevalent as the exposure increased (p<0.001), suggesting disturbance in cellular nutritional support essential for cellular motility. Prolonged liquefaction time, reduced semen volume and viscosity as well as altered seminal plasma protein, fructose and cholesterol level among the workers indicated dysfunction of accessory sex glands viz. prostate and seminal vesicle after occupational lead exposure (p<0.001). Deterioration of sperm count, structural abnormality of spermatozoa and sperm head DNA hyploidy was also associated with high blood and semen lead levels in the paint factory workers (p<0.001) without interfering serum FSH, LH and testosterone level (non-significant at p<0.05). Conclusion: Therefore, the present study suggested that at the present exposure level lead might cross blood-testis-barrier and increased its value in semen of the occupationally exposed paint factory workers in Bangalore, India, thereby producing detrimental effects on semen quality and sperm characteristics. Key words: Blood/semen lead level, cell cycle, paint factory workers, semen biochemistry, sperm morphology.     

剖宫产术后再次妊娠分娩方式的 Fisher 判别

目的：应用Fisher判别法归类剖宫产术后再次妊娠的分娩方式,为剖宫产史（ PCS）孕妇分娩方式的选择提供参考。方法以愿意阴道试产的216例 PCS 孕妇为研究对象,采集年龄、职业、临产前体重指数（ BMI）、距前次剖宫产时间、妊娠并发症、超声测量胎儿腹围、胎儿双顶径、子宫下段厚度、胎儿性别,记录分娩中Bishop宫颈成熟评分、宫缩剂应用情况等。结果①分娩方式：阴道自然分娩成功66例,阴道助产分娩成功58例,剖宫产分娩92例。②单因素分析显示：3种分娩方式在职业、临产前BMI、胎儿腹围、双顶径、宫缩剂应用、距前次剖宫产时间、子宫下段厚度和Bishop评分方面差异有统计学有意义,P＜0．05。③逐步法Fisher判别入选判别函数的因素是胎儿腹围（ X1）、子宫下段厚度（ X2）、Bishop评分（ X3）和应用宫缩剂（ X4）。 Fisher判别函数：Function 1＝-0．101X1＋2．202X2＋0．649X3＋0．289X4-9．243;Function 2＝0．258X1-0．156X2＋0.593X3＋0．409X4-14．384。④函数预测阴道顺产、阴道助产和剖宫产的敏感性分别是80．30％、56．90％和78．26％,总体符合率为73．15％。结论胎儿腹围、子宫下段厚度、Bishop评分和应用宫缩剂是影响PCS孕妇阴道分娩成败的重要因素。以此4项指标建立的Fisher判别函数可为PCS孕妇分娩方式的选择提供中等强度诊断效能。     

受精卵最佳显微注射时间的探索

目的探讨最佳显微注射受精卵的时间。方法对小鼠诱导超排卵后,在不同时间点采集受精卵,观察采集到受精卵的不同阶段中雄原核的状态,确定受精卵出现雄性原核的最佳时间。结果在注射人绒毛膜促性腺激素(HCG)后的23-25 h,出现雄性原核的受精卵所占的百分率达到75%以上,此时雌雄原核清晰可见,受精卵雄原核较大。结论在注射HCG后23-25 h的受精卵最适合进行显微注射。     

含雌激素的冷冻保护液对人精子冻贮效果的影响比较

[目的]探讨雌激素在各种冷冻保护液中对冻融后精子功能的影响。[方法]将正常人精液在4种不同冷冻保护液中进行冻贮。①单一甘油型冷冻保护液。②单一甘油型 雌激素冷冻保护液。③甘油混合型冷冻保护液。④甘油混合型 雌激素冷冻保护液。通过速冻法液氮冷冻24h后对照分析精子功能。[结果]①单一甘油型 雌激素冷冻保护液在精液冻融后精子的复苏率及活力较单一甘油型有明显提高(P<0.05)。②甘油混合型 雌激素冷冻保护液在精液冻融后精子的复苏率及活力较甘油混合型有明显提高(P<0.05)。[结论]雌激素加入人精液冷冻保护液中有提高精子功能的作用。     

Effects of microinjection of L-glutamate into locus coeruleus complex area on respiration

Summary The experiments were performed on 55 rabbits vagotomied, anesthetized with urethane and immobilized with flaxedil. Injection of L-glutamate (L-glu) into the locus coeruleus complex (Lc-Sc) area led to a marked increase in respiratory frequency (RF) and phrenic nerve discharge rate (phr. d) (16±3.5% and 6.5±2.5%;P<0.01 andP<0.05, respectively), a decrease in inspiratory and expiratory duration, and no obvious change in blood pressure (Bp) was observed. While prazosin (0.5 μg/μl), yohimbin (4 (μg/μl) and propranolol (2μg/μl) were preinjected into the nuclei tractus solitraii (NTS) respectively, the above-mentioned effects of L-glu on respiration was blocked. These results indicate that the excitation of neurons in the Lc-Sc can induce an increase in RF and phr d and the excitative effect of L-glu on RF was mediated by α₁,α₂ and β-receptors in the NTS, while the effect on phrd was mediated differently. This project was supported by grant no. 3870190 from the National Natural Sciences Foundation of China.     

人精液的冻贮研究

目的 研究在Ｔ－Ｇ型冷冻保护剂（Ｃｒｙｏｐｒｅｓｅｒｖａｔｉｖｅ ｍｅｄｉｕｍ,ＣＰＭ）作用下,速缓冻贮法对人精子结构和功能的影响。方法 随机双盲对照,将４５份正常精液随机分组,加与不加ＣＰＭ,以速缓冷冻法分别进行冻贮,冻贮前后分别进行常规分析、低渗肿胀试验、穿卵试验、超微结构观察及精子染色体分析。结果 未加ＣＰＭ组冻贮后的精液未见有精子复苏,经冷冻保存的精子质量明显降低,缓冻法对精子的影响显著小     

海马CA1区微量注射去甲肾上腺素对慢波睡眠的影响

目的探讨海马CA1区给予去甲肾上腺素（NA）对大鼠睡眠的影响。方法运用立体定位技术进行海马微量注射,通过多导睡眠描记（PSG）观察大鼠睡眠的变化。结果NA引起慢波睡眠（SWS）和总睡眠时间（TST）增加,NA的B型受体阻断剂普奈洛尔减少SWS和TST,并可阻断NA的促睡眠效应;NA的仅型受体阻断剂酚妥拉明对睡眠无影响,亦不能阻断NA的促睡眠效应。结论海马CA1区参与睡眠一觉醒的调节,海马中的NA通过B受体发挥其促进慢波睡眠的作用。     

未成熟卵母细胞体外成熟培养技术研究进展

<正>1935年,Pincus和Enzmanz[1]观察到兔的未成熟卵母细胞在体外培养条件下可以自发完成卵母细胞的成熟过程,自此在生殖领域引入了一个新的概念即卵母细胞体外成熟(in vitro maturation,IVM)。自1991年Cha[2]等报道IVM技术取得第1例妊娠以来,IVM技术已应用于治疗多囊卵巢综合征(PCOS)的不孕,预     

双绒毛膜三胎妊娠经阴道减胎术后妊娠结局分析

目的分析双绒毛膜三胎妊娠行减胎术后的妊娠结局。方法回顾性分析2007年1月至2011年1月在广西医科大学第三附属医院生殖中心经辅助生殖技术治疗受孕的20例多胎妊娠的临床资料,其中双绒毛膜三胎妊娠7例(A组),三绒毛膜三胎妊娠13例(B组)均于移植后33~56 d行减胎术,减灭1胎,保留2胎。比较两组自然胎儿丢失率、分娩率、婴儿畸形率。结果 A组的自然胎儿丢失率、婴儿畸形率显著高于B组(64.3%比19.2%,40.0%比0%),差异有统计学意义(P<0.05),两组分娩率(42.9%比83.3%)比较差异无统计学意义(P>0.05)。结论为了避免对妊娠结局的不良影响,双绒毛膜三胎妊娠减胎时宜减灭单绒毛膜双胎,保留单绒毛膜单胎。     

侧脑室微量注射去甲肾上腺素对大鼠应激性胃溃疡形成的影响

采用立体定位侧脑室微量注射去甲肾上腺素（ＮＡ）的方法，观察ＮＡ对冷束缚大鼠应激性胃溃疡形成的影响，并初步分析其作用的外周机制。结果表明：侧脑室注射ＮＡ可以明显减轻正常、中枢ＮＡ耗竭和幽门结扎大鼠的应激性胃粘膜损伤，而应激大鼠的胃液分泌量、总酸排出量和胃壁结合粘液量均不受ＮＡ注射的影响。提示中枢ＮＡ对大鼠应激性胃粘膜损伤具有保护作用，且这种作用与局部胃分泌功能变化无关。     

线粒体活性对人类精子活动力的影响

目的:分析线粒体活性对人类精子活动力的影响。方法:以线粒体特异性荧光染料——mitotracker deep red 633(MTDR对不同活力的精子进行染色,然后用流式细胞术测定精子线粒体活性(即MTDR荧光强度值)。用雷帕霉素(rapamycin,RP)和羟氯喹(hydroxychloroquineon,HCQ)调控精子活性水平,测定精子活动力的变化。结果:MTDR荧光强度值与前向运动速率(progressive motility,PR)不相关(r=0.182,P=0.268)。5 nmol/L和100 nmol/L的RP均可明显降低线粒体活性,50μmol/L HCQ可明显提高线粒体活性(P<0.05),但它们对精子活动力影响都不明显(P>0.05)。结论:线粒体活性对人类精子活力影响不明显。     

精子顶体酶活性检测方法研究

本文报告检测精子顶体酶活性的方法，是以Ｎａ一苯甲酰一ＤＬ一精氨酸一ｐ一硝酰基苯胺（ＢＮＰＮＡ）为底物，检测新鲜去精浆精子顶体酶活性。文中分析了实验的温度、时间、反应体系中精子浓度等因素对实验结果的影响，提出了最佳实验条件及注意事项。     

中缝背核注射对氯苯丙氨酸观察睡眠的变化及5-羟色胺神经元形态学的改变

目的观察大鼠中缝背核(DRN)微量注射对氯苯丙氨酸(PCPA)后睡眠的变化及5-羟色胺(5-HT)能神经元的形态学改变.方法采用核团微量注射、多导睡眠描记和免疫组织化学技术.结果DRN内微量注射PCPA(10μg)引起睡眠时间增加,尤以深慢波睡眠增加明显;免疫组织化学显示DRN内5-HT阳性神经元明显减少.结论大鼠DRN微量注射PCPA后睡眠增加且5-HT阳性神经元明显减少,提示5-HT神经元参与睡眠调节.