团头鲂卵泡细胞的超微结构

团头鲂排卵前的卵母细胞由外向内被鞘膜层基膜和颗粒层包围着。鞘膜层由鞘膜细胞和胶原纤维组成,鞘膜细胞的细胞质中充满了微丝。颗粒层由特殊颗粒细胞和颗粒细胞组成,特殊颗粒细胞的细胞质中含丰富的滑面内质网和具小管状内嵴的线粒体,表现出与类固醇激素合成有关的细胞器特征,颗粒细胞的细胞质中则含有发达的粗面内质网和具板层状内嵴的线粒体。     

原花青素对体外培养牛卵泡颗粒细胞增殖及其类固醇合成与分泌功能的影响

【目的】探究原花青素（proanthocyanidins,PC）对体外培养的牛卵泡颗粒细胞（granulosa cells,GCs）增殖及其类固醇合成与分泌功能的影响。【方法】采集性成熟荷斯坦奶牛卵巢组织,分离卵泡GCs,使用免疫荧光对其进行鉴定。用不同质量浓度（10,30,50,70和90 μg/mL）的PC对牛卵泡GCs作用不同时间（12,24和48 h）后,测定GCs存活率。以β-actin为内参基因,采用实时荧光定量PCR（qRT-PCR）检测10,50和90 μg/mL PC对牛卵泡GCs周期相关基因（CCND1和CCND2）、凋亡相关基因（caspase-3、caspase-9和Bim）以及类固醇激素合成相关基因（CYP19A1、CYP11A1、STAR和3β-HSD）相对表达量的影响,并检测培养上清液中类固醇激素（雌二醇（E₂）和孕酮（P₄））的浓度。【结果】分离获得了纯度较高的牛卵泡GCs。当PC质量浓度为50 μg/mL且培养24 h时,牛卵泡GCs的存活率最高。当PC质量浓度为50 μg/mL时,牛卵泡GCs周期相关基因CCND1和CCND2相对表达量均极显著升高（P＜0.01）;凋亡相关基因caspase-3和caspase-9相对表达量均极显著降低（P＜0.01）,Bim相对表达量显著降低（P＜0.05）;E₂和P₄浓度均极显著增加（P＜0.01）;类固醇激素合成相关基因CYP19A1、CYP11A1、STAR和3β-HSD相对表达量均极显著升高（P＜0.01）。当PC质量浓度为10 μg/mL时,牛卵泡GCs凋亡相关基因caspase-3的相对表达量显著降低（P＜0.05）;P₄浓度显著增加（P＜0.05）;CYP19A1和3β-HSD相对表达量均显著升高（P＜0.05）,CYP11A1和STAR相对表达量均极显著升高（P＜0.01）。当PC质量浓度为90 μg/mL时,E₂浓度显著增加（P＜0.05）,P₄浓度极显著增加(P＜0.01）;CYP19A1、CYP11A1、STAR和3β-HSD相对表达量均极显著升高（P＜0.01）。【结论】50 μg/mL PC通过促进牛卵泡GCs周期相关基因的表达和抑制凋亡相关基因的表达而促进牛卵泡GCs的增殖。10～90 μg/mL PC通过促进类固醇激素合成相关基因的表达而促进了牛卵泡GCs合成和分泌类固醇激素,进而影响牛卵泡GCs类固醇激素合成与分泌功能。     

Changes in the population of lymphocytes expressing CD4 and CD8 during the process of atresia of white follicles in hens

The aim of this study was to determine whether the population of lymphocytes expressing CD4 and CD8 molecules changed in the white follicles during atresia in chickens. Frozen sections of healthy, early atretic, advanced atretic and late atretic follicles were immunostained for CD4 and CD8, and the populations of positive cells were analyzed under a light microscope. In the healthy, early atretic and advanced atretic follicles both CD4+ and CD8+ cells were localized in the theca layer, but not in the granulosa layer. However, an influx of CD4+ and CD8+ cells was observed not only in the theca but also in the follicular cavity that was formed by disintegration of the oocyte in late atretic follicles. The frequency of CD4+ T cells in the theca layer did not differ among healthy, early atretic and advanced atretic follicles, but was significantly increased in the late atretic follicles (P < 0.05). The frequency of CD8+ cells showed a pattern of change that resembled that of CD4+ T cells, with a significantly greater population in late atretic follicles than the other follicles (P < 0.05). These results suggest that CD4+ and CD8+ cells are increased in the late atretic follicles, probably to promote the tissue regression.     

Surgical management of atresia ani and perineal hypospadias in a miniature donkey foal

A 3‐week‐old miniature donkey was evaluated for tenesmus and constipation resulting from atresia ani. The foal was also diagnosed with hypospadias extending from its perineum to the glans penis and caudoventral penile deviation. A common opening on the perineum allowed faeces and urine to be excreted together, preventing complete intestinal obstruction. Surgical correction of the atresia ani was performed, along with resection of the incomplete urethra, nonfunctional penis and testes. The anal and perineal reconstruction procedures resolved the miniature donkey's difficulty in defaecation. The anourethral portion of the repair healed, but the anoperineal portion dehisced and healed by second intention. The client was satisfied with the outcome.     

Developmental ability of oocytes retrieved from Meishan neonatal ovarian tissue grafted into nude mice

Ovarian xenografting makes it possible to obtain oocytes with fertilization ability from immature pigs of Western breeds. In this study, we applied these methods to the Meishan, an indigenous Chinese pig breed, and investigated the developmental competence of oocytes grown in their neonatal tissue after grafting into nude mice. First, mice harboring neonatal ovarian tissue were infused with follicle stimulating hormone (FSH) (62.5 U/ml) for 13 days starting at 10, 30, and 60 days after vaginal opening (D10‐, D30‐, and D60‐FSH groups, respectively). Development of antral follicles and their oocytes was most enhanced in the D60‐FSH group. For the next step, we examined the in vitro maturation ability of the oocytes recovered from host mice after infusion with FSH at a dose of 62.5 U/ml or 125 U/ml (FSH‐62.5 or ‐125 group) for 13 days starting at 60 days after vaginal opening. Many more oocytes with maturation ability were obtained from the FSH‐125 group. The FSH‐125 mature oocytes were fertilized in vitro, as shown by formation of male and female pronuclei, but did not reach the blastocyst stage. These results indicate that Meishan neonatal ovaries are able to produce oocytes with fertilization ability after being grafted into nude mice.     

Intrafollicular and systemic serotonin,oestradiol and progesterone concentrations in cycling mares

The hypothesis that a local serotonergic network might also exist in the follicle of mares remains poorly documented, with exception for humans and laboratory species. For this reason, the aim of the present study was to clarify this possibility, investigating intrafollicular serotonin concentrations of the cycling mare at ovulation time. Sixty ovaries collected from 30 clinically healthy mares of slaughterhouse meat production with clinically normal reproductive tracts after slaughtering were evaluated. Blood samples were taken prior to sacrifice. Follicles were classified in three categories in relation to size, as small (20–30 mm), medium (31–40 mm) and large (>41 mm), and the follicular fluid samples were extracted from each follicle. Intrafollicular and systemic serotonin (5‐HT), oestradiol‐17β (E₂) and progesterone (P₄) were determined by means of enzyme‐linked immunosorbent assay and RIA, respectively. Intrafollicular 5‐HT, E₂ and P₄ concentrations were higher than systemic ones (p < .05). 5‐HT concentrations increased in larger compared to medium follicles, without differences compared to small size follicles (p < .05). 5‐HT and E₂ (r = .79) and 5‐HT and P₄ (r = .79; p < .05) were positively correlated. 5‐HT and P₄ concentrations in follicular fluid increased progressively with the increase in follicular size (p < .05). Follicle diameter and E₂ (r = .85) and P₄ (r = .68) were correlated (p < .05). Since serotonin interacts with steroids, its role on steroidogenesis during growth of the dominant follicle may be suggested.     

氯化钴通过诱发DNA氧化损伤抑制猪卵泡颗粒细胞增殖的研究

旨在探究氯化钴（CoCl₂）诱导的DNA氧化损伤对猪卵泡颗粒细胞增殖的影响。本研究选取猪卵泡颗粒细胞作为试验材料,使用化学性低氧模型诱导剂CoCl₂处理猪卵泡颗粒细胞建立缺氧模型。前期研究已确定CoCl₂最适处理浓度,用200 μmol·L^-1 CoCl₂处理猪卵泡颗粒细胞12 h,将培养出的传代细胞分成4组处理,分别为：对照组、CoCl₂诱导缺氧处理组、GSH处理组、GSH+CoCl₂联合处理组。对照组为完全培养基培养12 h,CoCl₂诱导缺氧处理组给予终浓度200 μmol·L^-1 CoCl₂的培养基培养12 h,单独GSH处理组加入浓度为2 mmol·L^-1的GSH处理12 h,GSH+CoCl₂联合处理组加入200 μmol·L^-1 CoCl₂和2 mmol·L^-1的GSH联合处理细胞后培养12 h。12 h后检测各组细胞增殖活性、ROS水平、γH2AX蛋白活性水平和Oxo-8-G水平。采用CCK-8法检测其增殖活性;采用双氯荧光素（2’,7’-dichlorofluorescin diacetate,DCFH-DA）检测ROS水平;Western blot检测γH2AX蛋白活性水平,采用FITC荧光小鼠单克隆抗体检测Oxo-8-G水平。为了进一步明确CoCl₂诱导的颗粒细胞增殖阻滞与DNA氧化损伤的关系,本试验在CoCl₂处理的基础上添加抗氧化物GSH处理12 h后检测细胞增殖、ROS水平、DNA氧化损伤等相关指标。与对照组相比,200 μmol·L^-1 CoCl₂处理猪卵泡颗粒细胞后,细胞增殖活力极显著降低（P<0.000 1）,缺氧组ROS水平极显著升高（P<0.000 1）,γH2AX蛋白表达极显著升高（P<0.000 1）,Oxo-8-G水平极显著升高（P<0.000 1）。与缺氧组相比,在CoCl₂处理基础上添加GSH后,ROS、Oxo-8-G、γH2AX水平极显著降低（P<0.000 1）,并伴随细胞增殖活性的显著恢复（P<0.000 1）。CoCl₂可抑制猪卵泡颗粒细胞增殖活性,机制与诱导DNA氧化应激损伤有关。     

Integrated immunohistochemical and DNA copy number profiling analysis provides insight into the molecular pathogenesis of canine follicular lymphoma

Follicular lymphomas (FLs) typically exhibit a chromosome translocation that induces constitutive expression of the anti‐apoptotic bcl2 protein and accumulation of additional molecular defects. This rearrangement offers a promising therapeutic target, but its nature as a fundamental driver of FL pathogenesis remains unclear as 15% of cases lack the translocation. We performed an integrated immunohistochemical and genomic investigation of 10 naturally occurring FL cases from domestic dogs, showing that, as with human tumours, they exhibit marked heterogeneity in the frequency and intensity of bcl2 protein expression. Genomic copy number aberrations were infrequent and broadly consistent with those of other canine B‐cell lymphoma subtypes. None of the canine FL specimens exhibited a rearrangement consistent with the hallmark translocation of human FL, despite their remarkable histomorphologic similarity. Parallel exploration of canine and human cases may reveal alternative tumour‐initiating mechanisms other than BCL2 disruption, yielding a more complete definition of the molecular pathogenesis of FL.     

理囊散治疗卵泡囊肿不孕奶牛血清白介素与激素的相关性分析

探讨理囊散治疗卵泡囊肿不孕奶牛血液生殖激素和白介素含量关系,分析其相关性与回归方程.用ELISA法测定理囊散治疗卵泡囊肿不孕奶牛前后血清中促卵泡激素（FSH）、促黄体激素（LH）、孕激素（P）和雌二醇（E2）的浓度以及血清IL-1、IL-6含量,统计分析了4种激素和两种白介素间的相关关系与回归方程.结果表明,IL-1、IL-6与P呈正相关,与FSH、LH、E2为负相关.提示卵泡囊肿不孕奶牛血清中白介素与激素的分泌存在一定的相关性.