小鼠胚芽干细胞同种移植治疗慢性肝脏损伤的实验研究

将分离和体外扩增的小鼠胚芽干细胞,用BrdU标记后,经尾静脉同种移植于CCL4制备慢性肝损伤小鼠体内。分别于移植后2w、4w取出肝脏,进行连续冰冻切片,用免疫组织化学和免疫荧光双重染色检测移植细胞在受体小鼠肝内的存活、分布和肝细胞特异性白蛋白的表达;用PAS染色检测移植细胞的糖原代谢;HE染色观察移植动物肝组织的结构变化。探索胚芽干细胞在慢性损伤肝脏中的存活、分化和治疗作用。实验结果表明:胚芽干细胞经静脉移植后可以进入损伤肝脏中,并分化为肝样细胞和肝小叶内的其他细胞,并明显改善损伤肝脏的组织结构,具有较好的治疗作用。     

Advances in mammalian spermatogonial stem cell transplantation

Spermatogonial stem cell (SSC) transplantation is a novel technique by which testicular cells from normal, transgenic or mutant donor are introduced into the seminiferous tubules of recipient testes through microinjection. Subsequently, donor SSCs survive,migrate, anchor and proliferate in the recipient testis, furthermore, initiate spermatogenesis and even produce sperms capable of fertilization. This technique provides a new approach for the researches of spermatogenesis mechanism, regeneration of spermatogenesis in sterile individuals and reproduction of transgenic animals. This review focuses on the methodological breakthroughs and highlights the recent findings that have substantially increased understanding of SSC biology. The article provides a comprehensive overview of this technique and its multiple applications in basic science and medicine. And the perspective direction of this field in the near future is proposed.     

慢病毒载体转染犬睾丸生殖细胞

摘要: 目的观察慢病毒载体( Lentiviral vector,LV) 对犬生殖细胞的转染,为基于LV 的精子介导法制备转基因动 物提供依据。方法采用睾丸打点注射法,向比格犬两侧睾丸分别注入滴度为5 × 108、1 × 108、2 × 107 TU /mL 的 慢病毒液组成3 个剂量组,每点注射量为每只犬0. 2 mL。于注射后第2 周开始采集精液,通过精液DNA 的PCR 检 测和精子荧光镜检评估绿色荧光蛋白( green fluorescent protein,GFP) 基因的整合及表达。结果1) 在高剂量组,整 合并表达GFP 基因的精子于第4 周首现并持续至第17 周; 在中、低剂量组于第5 周首现,分别持续到第14 周、12 周。2) GFP 表达的高峰期在注射后第7 周～ 10 周。3) GFP 表达于整个精子,以顶体后区和精子尾颈部表达最强。 4) 注射后第2 周～ 6 周,中剂量组犬采精困难,高低剂量组犬精子畸形率有上升的趋势。5) 在GFP 表达的高峰期, 绿色荧光精子的百分率在高、中、低剂量组分别为43. 33% 、35. 53% 和4. 55% ,具有明显的差异。结论基于LV 的 精子介导转基因法( Sperm-mediated gene transfer,SMGT) 可成功获得转基因犬精子,转基因阳性率、表达的强度与 持续时间与慢病毒滴度相关。     

Neural stem cell transplantation in the repair of spinal cord injury

Neural stem cells are a pronising candidate for neural transplantation aimed at neural cell replacement and repair of the damaged host central nervous system (CNS). Recent studies using neural stem cells have shown that implanted neural stem cells can effectively incorporate into the damaged CNS and differentiate into neurons, astrocytes, and oligodendrocytes. The recent explosion in the field of neural stem cell research has provided insight into the inductive factors influencing neural stem cell differentiation and may yield potential therapies for several neurological disorders, including spinal cord injury. In this review, we summarize recent studies involving neural stem cell biology in both rodents and humans. We also discuss unique advantages and possible mechanisms of using neural stem cell trans plantation in the repair of spinal cord injury.     

鸭胚胎血液中PGCs数量及其外源DNA的体内转染

抽取了麻鸭和骡鸭胚胎发育第14－20期的血液，制作血涂片，PAS染色，观察血液中的原生殖细胞的含量，并对各个时期血液中原生殖细胞的含量进行了统计，最高含量都出现在第16期,分别为8．9752％和2．1871％，用In vivo Gene Delivery Systern和pEGFP-N对PGCs进行了外源DNA的体内转染。结果在6只胚胎中的3只中发现GFP报告基因的表达。     

自体造血干细胞移植后的胸腺近期输出功能

目的:了解血液病患者接受自体造血干细胞移植(auto-HSCT)后外周血中的T细胞受体重排删除环(TRECs)水平,从而了解其胸腺近期输出功能和T细胞免疫重建情况。方法:采用实时定量PCR法检测9例血液病患者经auto-HSCT治疗后外周血单个核细胞(PBMNCs)中的TRECs水平;22例正常人外周血样本作为对照。结果:PBMNCs中的TRECs水平有着较大的个体差异,年龄与PBMNCs中的TRECs水平呈负相关;auto-HSCT组的每1 000个PBMNCs中的TRECs平均拷贝数(2.533±4.194)明显低于正常组(4.367±3.626)(P<0.05),但部分病例的TRECs水平已基本达到正常水平。结论:在一定时间内,auto-HSCT后的胸腺近期输出功能仍较低,不同个体T细胞免疫重建时间存在差异。     

5种细胞周期调控基因在小鼠生殖细胞发育过程中的表达研究

 以成熟(10周龄以上)的昆明种正常小鼠的精巢和卵巢为材料,利用地高辛标记的基因探针进行组织切片上的DNA-mRNA分子原位杂交,研究了PCNA,cdc2,cyclin D1,p2 1和 p16 5种细胞周期调控基因在生殖细胞发育过程中的表达.结果表明:PCNA基因在睾丸组织的精原细胞和精母细胞中有强杂交信号,而在雌性生殖细胞及滤泡细胞的发育过程都没有杂交信号;cyclin D1,cdc2,p2 1,p16基因在生殖细胞的发育过程中都没有,表明这些基因并没有参与小鼠生殖细胞的生长和分化调控.这些事实表明在生殖细胞发育过程中,控制细胞增殖和增殖抑制的基因与培养细胞有不同的机制,它们可能采用了不同的调控系统.     

实验用五指山小型猪胚胎生殖细胞的分离培养

目的对从猪的原始生殖细胞分离获得胚胎干细胞的方法进行初步研究。方法胎儿取自怀孕26～28d的实验用五指山母猪。分离胎儿生殖嵴获得PGCs,接种于STO饲养层上,以DMEM+2mmol/L谷氨酰胺+0.1mmol/L非必需氨基酸+0.1mmol/Lβ-巯基乙醇+100IU/mL青霉素+100μg/mL链霉素+15%FCS作为培养基,对细胞进行培养传代。结果培养的细胞传至4～5代,形态多样,经AKP染色、SSEA-1免疫荧光染色、Oct-4免疫荧光标记等方法鉴定为阳性,具有胚胎干细胞的特征。     

超声监测卵泡在不孕症中的应用

目的探讨应用超声监测卵泡发育，指导临床治疗不孕症。方法对本院门诊妇科就诊的78例不孕症患者，采用超声对用促排卵药前、后卵泡进行定期观察。结果可明显观察到用促排卵药前、后卵泡发育情况。结论采用超声检查可动态观察卵泡发育及排卵，有利于指导临床妇科对不孕症的治疗。     

无臭蒜素与玉米胚芽油的制取及其在化妆品中的应用

论述了无臭蒜素与玉米胚芽油的制取,探讨了无臭蒜素与玉米胚芽油的保健功效及其在化妆品中的应用。     

L型菌所致肺部感染性疾病的研究

目的:探讨提高培养L型菌检出率的方法,了解L型菌对常用抗生素的耐药谱,为防治L型菌所致的肺部感染提供检测依据.方法:对200例肺部感染患者痰标本进行常规与L型菌培养的检测,对分离培养出的细菌进行药敏试验.结果:分离出细菌39株,其中L型菌21株,占53.8%,普通细菌18株,占46.2%;L型菌以克雷伯茵和大肠埃希氏菌最多,各占42.8%.药敏试验对丁胺卡那霉素、庆大霉素、左氧氟沙星及部分头孢类抗生素较为敏感,对氨苄青霉素耐药.结论:L型菌在肺部感染性疾病中比例较高,应将L型菌检测作为常规实验开展,以减少漏诊.同时,临床合理使用抗生素能减少L型菌的产生及提高治疗效果.     

脱脂麦胚蛋白粉的功能特性及其应用研究进展

作为面粉工业的副产品,小麦胚芽的世界年蕴藏量约１７００万吨,大部分未被开发利用小麦胚芽具有十分丰富的营养素,主要是蛋白质、脂肪、维生素、矿物质和膳食纤维,还含有谷胱甘肽等生理活性物质经脱脂的麦胚蛋白粉具有优良的功能特性,如溶解度、持水能力、保脂能力、起泡能力、乳化特性和胶凝作用等在食品中添加麦胚蛋白粉,不仅可以强化食品营养,还可以改善食品的结构和品质,提高产品的得率国外麦胚粉主要用于法兰克福香肠研究结果表明,麦胚蛋白粉加入量不超过５．０％时,产品营养构成更合理,对外观和感官品质影响不大,产品得率增加此外,麦胚蛋白粉还用于松饼、面包、饼干等食品中,或制成乳化蛋白饮料等目前,对小麦胚芽蛋白的研究与大豆蛋白和玉米胚芽蛋白相比,还比较粗浅,尤其在麦胚蛋白的保脂能力、蛋白分离物和蛋白浓缩物的功能特性以及在其它食品中的应用等方面有待进一步加强     

双螺杆挤压加工麦胚芽冲剂的研究

以感官评价为指标,通过挤压膨化、粉碎和调配等工艺研制了麦胚芽和玉米粉混合的冲剂产品。结果表明：麦胚芽经挤压加工后粗纤维含量下降达46．2％,水溶性蛋白质含量从原来的3．6mg／g增加到38．1mg／g。配料中玉米粉含量越高,挤压加工产物的膨化率越高,但不能改善其粉碎品的溶解性。物料配比对麦胚芽冲剂产品的感官评分有明显的影响（p〈0．1）。多重比较结果表明：物料配比A3（麦胚芽：玉米粉=9：1）与A2（麦胚芽：玉米粉=7：3）之间麦胚芽冲剂的感官评分无明硅的差异。     

小麦胚芽复合营养粉配方及工艺研究

以小麦胚芽、大米和奶粉为主要原料,通过配方及喷雾干燥工艺研究,得出了产品的基本配方为：小麦胚芽为200 g,大米为200 g,奶粉为100 g;经分析,喷雾干燥工艺参数对产品冲调性有较大影响,当进风温度为180℃,喷雾压力为1.0 MPa,物料浓度为30%时,所制得的产品具有良好的感官品质及冲调性能.     

人脐血间充质干细胞移植治疗大鼠急性肝损伤

目的:探讨人脐血间充质干细胞对急性肝损伤大鼠模型的治疗作用.方法:体外培养人脐血间充质干细胞.60只SD大鼠随机分为3组,各20只:空白对照组、模型对照组、治疗+模型组.模型对照组大鼠采用体积分数50%四氯化碳橄榄油溶液腹腔注射造成急性肝损伤疾病模型,测定ALT/AST水平、肝脏切片HE染色评估造模效果.治疗+模型组大...     

静宁鸡原生殖细胞的分离及形态学观察

原生殖细胞(PGCs)是配子的始祖细胞,它可发育演变生成卵细胞或精子细胞.上世纪90年代发展了体外培养PGCs技术,现在PGCs已能通过体外培养形成多潜能干细胞(ES),对于研究体外分化机制和细胞治疗具有重要意义.文章通过对静宁鸡种蛋进行36~82 h孵化,并对其血涂片经PAS染色后,在显微镜下观察其原生殖细胞(PGCs)的形态结构,以便为鸡的胚胎干细胞分离培养奠定基础.实验结果:原生殖细胞呈圆形或卵圆形,明显较其他细胞大2~4倍,PGCs的细胞核颜色较深,偏向一侧,后期常聚成一团.     

Effect of kinase-negativeEGFR gene on differentiation of embryonic germ cell line EG4

EG4 cells derived from primordial germ cells (PGCs) of 10.5 d post coitum 129/svJ mouse embryos can be used as a model system for in vitro differentiation study due to their pluripotential development ability. EG4 cell lines with stable expression of kinase-negative EGFR cDNA, designated EG4-EGFR^d, were generated by gene transfection. We found that: (ⅰ) EG4-EGFR^d cells share the similar morphology and growing character with wildtype cells that can maintain undifferentiated state in long term culture. (ⅱ) Treatment of EG4 cells with RA resulted in differentiation of adipocyte, while in mutant clones of EG4-EGFR^d, adipocytes were sparse or absent under the same condition, indicating the role of EGFR expressed during adipocyte development. (ⅲ) Histological analysis showed that predominant tissues in teratocarcinomas derived from EG4-EGFR^d cells and wildtype cells are different. A large amount of undifferentiated cells was present in those coming from mutant cell clones. In addition some cardiac and skeletal muscles are prominently differentiated cell types. EG4 wildtype cells produced multiple differentiated cell types of three primary germ layers such as cartilage, epithelia and neural tube. These studies suggested that EGFR-dependent differentiation was inhibited in kinase-negative EG4 cells.     

Abnormal expression of centrosome protein (centrin) in spermatozoa of male human infertility

To study the relations between male infertility and centrosome protein (centrin) and the functions of centrin in spermatogenesis, the matured spermatozoa of 10 normal male people and 18 male infertility patients were stained by immunofluorescence labeling antibody against centrin. The results showed that two fluorescence signal dots appeared in the normal male spermatozoa and were located at the base of flagellum. They are proximal centriole and distal centriole. However, in some spermatozoa of the male infertility, centrin protein was located abnormally at the base of flagellum and its staining signals were spread, the normal proximal and distal centrioles were confused and could not be recognized separately. These results suggest that abnormality of centrosome protein may be related to male infertility. This discovery may be used as a marker of abnormal sperm and male infertility.     

不育不孕症生殖道衣原体及支原体的感染

目的：了解不育不孕症生殖道衣原体及支原体的感染及其与年龄、性别、原发性、继发性、不洁性接触史的关系;解脲支原体（ＵＵ）感染、生殖道炎症与精子活动率、抗精子抗体（ＡｓＡｂ）的关系,进一步探讨不育不孕的致病因素,提高诊断准确率和治愈率。方法：选用衣原体检测及支原体培养技术、抗精子抗体（ＡｓＡｂ）检测技术、明胶凝集试验（Ｇｅｌａｔｉｎｇ Ａｇｇｌｕｔｉｎａｔｉｏｎ Ｔｅｓｔ,ＧＡＴ）,试管玻片凝集试验（     

Replacement of inner cell mass in mouse

The intra- or inter-strain reconstituted blastocysts were produced by replacing the inner cell mass of Kunming mouse blastocysts with that of Kunming or C57BL/6 mouse strain blastocysts. A total of 192 intra-strain reconstituted blastocysts were transferred into 17 pseudopregnant Kunming mice, and 2 reconstituted embryos were developed into term: while 115 inter-strain reconstituted blastocysts were produced, analysis of the reconstituted blastocysts showed that the morphology and cytoskeleton srtucture of the blastocysts were not different from those of normal blastocysts, however, no viable offspring was obtained after embryo transfer for these inter-strain reconstituted blastocysts. The results demonstrated that the intra-strain reconstituted blastocysts could normally develop into term, whereas the inter-strain reconstituted blastocysts possessed less developmental potential as the intra-strain reconstituted blastocysts. This study may give light to solve the problem of low implantation rate and placenta abnormality in mammal cloning.