激光对小鼠精母细胞联会复合体影响的研究

以亚离子和He-Ne激光照射活体雄性小鼠睾丸，观察小鼠睾丸中精母细胞的联会复合体。发现小鼠精母细胞的联会复合体发生一定程度的畸变，说明激光辐射不仅可造成哺乳动物表面组织热损伤，还有可能诱发染色体变异，这对今后激光在遗传育种方面的进一步开发应用有重要意义。     

PCNA基因在小鼠睾丸发育过程中的表达研究

为探讨PCNA基因在小鼠睾丸生后发育全过程中的表达规律,以16组出生后不同发育时期的昆明种正常小鼠睾丸组织为材料,在其石蜡组织切片上进行PCNA蛋白的免疫组化检测及PCNA基因的DNA-mRNA原位杂交检测.结果发现:PCNA蛋白仅在精原细胞及初级精母细胞中表达,其表达峰值出现在小鼠生后第11天;PCNA基因在初级精母细胞、次级精母细胞、圆形精子细胞以及少数精原细胞中均有表达,其中初级精母细胞中的表达信号相对强烈.上述结果表明,PCNA基因参与了小鼠睾丸发育过程中精原细胞及初级精母细胞的DNA复制过程,其mRNA表达和蛋白质表达存在着一定差异.     

X射线的直接照射与间接照射对猕猴(Macaca mulatta)生殖细胞的影响

雄性猕猴8只分成对照一直接照射,对照-间接照射,直接照射-间接照射,先阉割-后阉割四个组。每组2只猴子,分别给以200仑X射线直接照射或间接照射——睾丸用6毫米厚的铅板保护后照射躯体其余部分。实验结果指出直接照射引起精原细胞,精母细胞减少。间接照射后,A型精原细胞减少,精原细胞总数有减少趋势,B型精原细胞及精母细胞数量均有变动但似无一定的规律,畸变细胞数目没有增多。这些表明间接照射可能产生了某种物质影响了细田胞的正常代谢活动,可是还无法定量地反映这种间接辐射效应。     

Ag3PO4/ZnO复合物的制备及其降解甲基橙的研究

采用银氨辅助法合成了Ag_3PO_4,超声波分散法制备Ag_3PO_4/ZnO复合物,利用X-射线衍射(XRD)和扫描电镜(SEM)对催化剂进行了表征.在超声照射下,讨论了煅烧温度、煅烧时间、照射时间和染料浓度等对甲基橙降解率的影响.结果表明:Ag_3PO_4/ZnO在煅烧温度为450°C,煅烧时间为60min,超声照射180min时,甲基橙降解率可达85.36%,高于纯ZnO时的降解率.     

小鼠睾丸TERT表达的增龄变化

应用免疫组织化学和图像分析方法研究了10,20,30,60,90天龄的小鼠睾丸不同分化类型生殖细胞中端粒酶催化亚基TERT的表达.结果发现在精子发生过程中精原细胞及成熟精子中均未见TERT阳性表达,而初级精母细胞尤其是粗线期精母细胞和精子细胞是高表达TERT的细胞类群,附睾管上皮细胞也呈TERT阳性表达;同时TERT在小鼠个体发育过程中的表达存在着从弱到强再到弱的动态变化趋势.结果表明TERT在小鼠睾丸中的表达特点与精子发生过程密切相关,可能对维持生殖细胞遗传物质的完整性和减数分裂的正常进行发挥着重要作用.     

小鼠睾丸发育全过程的组织学观察

为系统观察小鼠睾丸组织发育过程,将生后1～57d处于不同发育时期的17组昆明种正常小鼠睾丸组织制备石蜡切片、进行H.E染色分析。结果表明:小鼠生后初期睾丸曲细精管中只有支持细胞和原始生精细胞;至生后8d时出现B型精原细胞;15d时出现初级精母细胞;23d时出现次级精母细胞及少量圆形精子细胞,此后圆形精子细胞逐渐增多;30d时圆形精子细胞发生变态;36d时大量精子开始稳定出现于曲细精管管腔中,并延续至此后各期。这意味着小鼠生精细胞在出生后经过一个相对连续的发育分化过程,至生后约36d发育成熟。这为细化小鼠睾丸组织发育和精子发生过程提供了详细资料。     

松花粉对小鼠睾丸染色体的影响

用赤松粉对小白鼠精母细胞的染色体进行了致畸变的测试,表明赤松花粉在ＬＤ〉１３０００ｍｇ／ｋｇ的情况下无致染色体畸变作用。     

纳米氧化锌(ZnO)催化超声波降解酸性红B的研究

采用高温活化处理过的纳米氧化锌(ZnO)粉末为催化剂,研究了超声照射对酸性红B的降解.另外,还考查了超声照射时间,酸性红B初始浓度和纳米ZnO的添加量对酸性红B降解反应的影响.结果表明:在纳米ZnO作用下,酸性红B的超声降解效果明显优于单纯超声波照射降解.降解动力学符合一级反应.在超声波频率40 kHz,输出功率50W,催化剂用量1.0 g/L,pH为7.0,温度25℃,酸性红B水溶液初始浓度10 mg/L的条件下,照射120min左右时降解率即可达到86%以上.     

黄犏牛与黄牛b-Boule基因编码区序列、结构、表达模式和睾丸组织表达水平的差异分析

Boule是动物精母细胞减数分裂过程中的必需蛋白,与精子发生减数分裂阻滞、雄性不育等密切相关.文中通过PCR扩增和克隆测序获得黄犏牛b-Boule基因的cDNA全序列,生物信息学分析发现黄犏牛b-Boule基因编码区全长885 bp,具有典型的RNA识别基序(RRM)和DAZ重复序列;与黄牛氨基酸序列的同源性为98.65%,仅在C-末端有4个氨基酸的差异,典型结构域的氨基酸序列和高级结构相同.RT-PCR发现b-Boule基因在黄牛和黄犏牛睾丸组织中特异表达,原位杂交显示b-Boule存在于初级精母细胞和次级精母细胞的细胞质中.荧光定量PCR发现黄牛睾丸组织中b-Boule基因表达水平显著高于黄犏牛(P<0.05),且黄犏牛精子发生减数分裂阻滞的表型与人、小鼠BDule基因表达缺失或降低引起的不育表型一致,因此推测b-Boule基因可能在精母细胞减数分裂过程中发挥重要作用,且与黄犏牛雄性不育有密切的关系.     

X射线一次急性照射与分次累积照射对猕猴精子发生中染色体畸变的影响

本实验以性成熟猕猴(Macaca mulatta)12只,分为对照、一次急性照射和分次累积照射三组,每组4只猴子。经200仑X 射线一次急性和分次累积(每隔24小时照射20仑,10天内累积总剂量为200仑)局部照射猕猴睾丸,以精子发生中精母细胞第一次减数分裂后期和早末期所出现的染色体畸变为指标,此较研究猕猴在总剂量和剂量强度相同的情况下,经一次急性照射和分次累积照射的细胞遗传学效应。对照组的平均染色体畸变率为0.297±0.103%,200仑X 射线一次急性照射组和分次累积照射组的平均染色体畸变率分剐为2.979±0.179%和1.276±0.144%。由此可见,两种不同处理组的染色体畸变率均高于对照组,而一次急性照射组的染色体畸变率又比分次累积照射组增加一倍以上。经统计分析指出:各组染色体畸变率之间差异是显著的,对照～处理P<0.001(对照～一次急性照射P<0.001;对照～分次累积照射P=0.005-0.001),一次急性照射～分次累积照射P=0.005-0.001。实验证明:在总剂量和剂量强度相同时,猕猴睾丸经X 射线一次急性照射所引起的细胞遗传学损伤比分次累积照射更为严重,并表明200仑X 射线分次累积照射诱发染色体畸变的效应并不是按照简单的相加法则而累积的。     

小鼠精原细胞体外培养分化的研究

目的 探讨小鼠精原细胞的分离纯化及体外培养条件。方法采用组合酶消化法分离纯化小鼠精原细胞和支持细胞,在添加FSHC和TC的基本培养基中进行体外培养。采用反转录PCR方法检测精子细胞特异性基因过渡蛋白9（Tnp2）的表达以对分化细胞进行鉴定。结果培养5d后有分化的圆形精子细胞出现,培养7d后收集的细胞可见Tnp2基因表达。结论通过体外培养小鼠精原细胞可迅速完成减数分裂过程和减数分裂后的分化过程,并可获得更接近成熟阶段的精子细胞。     

Germ cell transplantation in infertility mouse

This work investigated the spermatogenesis in an infertility BALB/c-nu mouse model by reinfusing germline stem cells into seminiferous tubules. Donor germ cells were isolated from male FVB/NJ-GFP trensgenic mice. Seminiferous tubule microinjection was applied to achieve intratubular germ cell transfer. The germ cells were injected into exposed testes of the infertility mice. We used green fluorescence and DNA analysis of donor cells from GFP transgenic mice as genetic marker. The natural mating and Southern blot methods were applied to analyze the effect of sperm cell transplantation and the sperm function after seminiferous tubule microinjection. The spermatogenesis was morphologically observed from the seminiferous tubules in 41/60 （68.33%） of the injected recipient mice using allogeneic donor cells. In the colonized testes, matured spermatozoa were seen in the lumen of the seminiferous tubules. In this research, BALB/c-nu infertility mouse model, the recipient animal, was used to avoid immunological rejection of donor cells, and germ cell transplantation was applied to overcome infertility caused by busulfan treatment. These results demonstrate that this technique of germ cell transplantation is of great use. Germ cell transplantation could be potentially valuable to oncological patients.     

人参皂甙Rb1和Rg1对小鼠睾丸支持细胞增殖的影响

观察人参皂甙Rb1(ginsenodde Rb1,GRb1)和Rg1(ginsenoside Rg1,GRg1)对体外培养的小鼠睾丸支持细胞(sertoli cell)增殖的影响,探讨二者的药理作用机制.取原代培养传代一次第3天的支持细胞,加入不同浓度的GRb1和GRg1,利用MTT比色分析法和BrdU增殖细胞标记法,检测不同浓度GRb1和GRg1对体外培养支持细胞增殖的影响.结果显示,20 mg/L GRb1对支持细胞增殖有明显促进作用,而不同浓度的GRg1对支持细胞的增殖无明显影响.GRb1能维持小鼠支持细胞的存活,并在适当浓度范围内可以明显促进支持细胞的增殖,而GRg1对支持细胞的增殖无明显影响.     

超声激活血卟啉对S180瘤细胞杀伤作用的观察

光敏剂血卟啉具有在肿瘤组织易聚集的特性,受激发时发生光敏反应,产生具有细胞毒性的单线态氧,它对癌细胞的显微结构具有一定的破坏作用,本文报道采用超声激活血卟啉的方法,探讨超声激活血卟啉对S180瘤细胞形态结构的影响,为超声化学激活血卟啉抗癌研究提供形态学依据。     

Zfy gene expression patterns are not compatible with a primary role in mouse sex determination

The Y chromosome determines maleness in mammals. A Y chromosome-linked gene diverts the indifferent embryonic gonad from the default ovarian pathway in favour of testis differentiation, initiating male development. Study of this basic developmental switch requires the isolation of the testis-determining gene, termed TDF in humans and Tdy in mice. ZFY, a candidate gene for TDF, potentially encodes a zinc-finger protein, and has two Y-linked homologues, Zfy-1 and Zfy-2, in mice. Although ZFY, Zfy-1 and Zfy-2 seem to map to the sex-determining regions of the human and mouse Y chromosomes, there is no direct evidence that these genes are involved in testis determination. We report here that Zfy-1 but not Zfy-2 is expressed in differentiating embryonic mouse testes. Neither gene, however, is expressed in We/We mutant embryonic testes which lack germ cells. These observations exclude both Zfy-1 and Zfy-2 as candidates for the mouse testis-determining gene.     

Sperm from neonatal mammalian testes grafted in mice

Spermatogenesis is a productive and highly organized process that generates virtually unlimited numbers of sperm during adulthood. Continuous proliferation and differentiation of germ cells occur in a delicate balance with other testicular compartments, especially the supporting Sertoli cells. Many complex aspects of testis function in humans and large animals have remained elusive because of a lack of suitable in vitro or in vivo models. Germ cell transplantation has produced complete donor-derived spermatogenesis in rodents but not in other mammalian species. Production of sperm in grafted tissue from immature mammalian testes and across species has not yet been accomplished. Here we report the establishment of complete spermatogenesis by grafting testis tissue from newborn mice, pigs or goats into mouse hosts. This approach maintains structural integrity and provides the accessibility that is essential for studying and manipulating the function of testes and for preserving the male germ line. Our results indicate that this approach is applicable to diverse mammalian species.     

小鼠腹腔注射阿霉素诱导骨髓细胞微核率的最佳时间—剂量效应分析

目的：寻找一次染毒条件下,腹腔注射阿霉素诱导小鼠骨髓细胞微核率出现峰值的时间-剂量条件。方法：将150只小鼠随机分为5组,每组30只,雌雄各半。腹腔注射,1次给药,对照组给生理盐水,处理组的阿霉素剂量分别是2.5 mg/kg、5 mg/kg、10 mg/kg和20 mg/kg。染毒18 h、24 h、30 h、48 h和72 h后取小鼠骨髓进行微核分析,评价阿霉素对小鼠的遗传毒性作用。结果：理想的腹腔注射诱导的小鼠骨髓细胞微核率峰值的条件就是：一次染毒,阿霉素剂量在10%LD50即2.5 mg/kg至80%LD50即20 mg/kg的范围内剂量越大微核率越高,取样时间在30~48 h间微核率最高。     

超声协同钙浸渍对樱桃番茄贮藏过程中番茄红素和品质特性的影响

通过研究超声协同钙浸渍处理对樱桃番茄贮藏过程中理化品质和番茄红素的影响,阐明了超声协同钙浸渍提高樱桃番茄品质特性的原因。采用质构仪、色差计和高效液相色谱仪研究了樱桃番茄贮藏过程中硬度、色泽和番茄红素含量的变化,同时利用核磁共振技术分析了樱桃番茄贮藏过程中番茄红素结构特性的变化。研究结果表明:超声协同钙浸渍处理可促进钙离子渗入到果肉组织中,有效延缓贮藏期间樱桃番茄硬度的下降,同时抑制果实褐变指数、失重率和腐烂率的上升,维持番茄果实较高的可滴定酸和维生素C含量;与对照组相比,超声协同钙浸渍对樱桃番茄全反式番茄红素含量无显著性影响。核磁共振结果表明,超声协同钙浸渍处理有效抑制了樱桃番茄贮藏期间全反式番茄红素、5-顺式番茄红素和9-顺式番茄红素的氧化,维持了其结构完整性。超声协同钙浸渍技术可较好地维持樱桃番茄贮藏期间的营养和品质,研究结果旨在为超声协同钙浸渍技术在果蔬保鲜中的应用提供理论依据。     

Spermatogenic failure in male mice lacking H-Y antigen

The mammalian Y chromosome carries a factor that initiates male sexual development by directing the fetal gonads to form testes. Wachtel and his colleagues proposed that this testis-determining function of the Y is mediated by the male-specific cell-surface antigen H-Y, originally defined by skin grafting. This attractive hypothesis, which has been widely accepted, was based on the assumption that serological tests using antisera raised against male cells were recognizing H-Y antigen. Although disputed this assumption is supported by some recent studies. However, mice have been described which develop testes but lack the cell-surface H-Y antigen as defined by T-cell-mediated transplantation tests. Thus, although it remains possible that a serologically detected male-specific antigen is responsible for testis determination, it seems that H-Y, as originally defined, is not. We show here that H-Y negative male mice, in losing the genetic information that encodes H-Y, have also lost genetic information required for spermatogenesis. This result identifies a gene on the mouse Y, distinct from the testis-determining gene, which is necessary for spermatogenesis, and raises the intriguing possibility that the product of this 'spermatogenesis gene' is H-Y antigen.     

卵巢切除小鼠冠状动脉封堵诱导心肌梗塞模型的建立与评估

摘要:目的 建立模拟绝经后妇女发生心肌梗塞( MI) 的小鼠模型。 方法 将雌性小鼠卵巢切除( OVX) 后进行左冠状动脉前降支封堵诱导心肌梗塞发生;通过超声、血流动力学、形态学分析、免疫组织化学和 TUNEL 染色评估小鼠心脏功能和心肌梗塞面积的变化及心肌纤维化的程度。 结果 卵巢切除联合心肌梗塞可加重心脏功能受损,显著增加心肌纤维化和心肌梗塞面积,可诱导细胞凋亡和 TGF-β1 表达,血清中促炎细胞因子 TNF-α 和 IL-10 水平显著增加。 结论 成功建立模拟绝经后女性发生 MI 的小鼠模型,并对其功能和组织学特征进行评估。