高血糖对小鼠卵母细胞Ca2+振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体表达的影响

目的 探讨高血糖对小鼠卵母细胞Ca2+振荡和卵泡颗粒细胞肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达的影响.方法 出生后20d ICR雌鼠,用链脲酶素建立急性高血糖模型;高血糖模型雌鼠及正常雌鼠注射10IU/只孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(hCG)超排卵,注射hCG后0h、2h、6h和10h取生发泡(GV)期卵母细胞,双光子激光扫描共焦显微镜检测GV期卵母细胞的Ca2+振荡;hCG注射后6h、10h取卵丘-卵母细胞复合体(COC),利用免疫荧光、Western blotting检测TRAIL的表达.结果 1.高血糖组卵母细胞Ca2+振荡频率高于正常对照组,于hCG注射后2h、6h差异显著(P<0.05);2.免疫荧光、Western blotting蛋白半定量检测显示,高血糖组颗粒细胞TRAIL表达量高于正常对照组,于hCG注射后6h、10h差异有显著性(P<0.05).结论 高血糖加快小鼠未成熟卵母细胞Ca2+振荡的频率;高血糖促进小鼠卵泡颗粒细胞TRAIL的表达.     

Molecular determinants on the insect sodium channel for the specific action of type II pyrethroid insecticides

Pyrethroid insecticides are classified as type I or type II based on their distinct symptomology and effects on sodium channel gating. Structurally, type II pyrethroids possess an α-cyano group at the phenylbenzyl alcohol position, which is lacking in type I pyrethroids. Both type I and type II pyrethroids inhibit deactivation consequently prolonging the opening of sodium channels. However, type II pyrethroids inhibit the deactivation of sodium channels to a greater extent than type I pyrethroids inducing much slower decaying of tail currents upon repolarization. The molecular basis of a type II-specific action, however, is not known. Here we report the identification of a residue G¹¹¹¹ and two positively charged lysines immediately downstream of G¹¹¹¹ in the intracellular linker connecting domains II and III of the cockroach sodium channel that are specifically involved in the action of type II pyrethroids, but not in the action of type I pyrethroids. Deletion of G¹¹¹¹, a consequence of alternative splicing, reduced the sodium channel sensitivity to type II pyrethroids, but had no effect on channel sensitivity to type I pyrethroids. Interestingly, charge neutralization or charge reversal of two positively charged lysines (Ks) downstream of G¹¹¹¹ had a similar effect. These results provide the molecular insight into the type II-specific interaction of pyrethroids with the sodium channel at the molecular level.     

卵巢储备功能的评价

卵巢储备功能,又称卵巢储备（ovarian reserve）,是指卵巢产生卵子数量和质量的潜能,间接反映卵巢的功能。据估计约10％的不孕妇女罹患卵巢储备功能降低。目前卵巢储备功能的检测方法有多种,它与辅助生殖技术（ART）时卵巢反应性的关系已进行了较多的研究,而与妊娠率、分娩率的相关性则研究较少,尚无公认的最佳的卵巢储备检测方法,多项指标结合应用检测卵巢储备能获得更好的效果。探讨卵巢储备的检测方法及其在临床的应用,对指导妇女在卵巢储备明显下降前受孕和治疗不孕症,有重要的临床价值。     

Effect of bovine cumulus–oocyte complexes-conditioned medium on in-vitro maturation of canine oocytes

Purpose

To investigate the ability of medium conditioned with bovine cumulus–oocyte complexes (COCs) to support nuclear maturation of canine oocytes recovered from domestic dog ovaries.

Methods

Cumulus–oocyte complexes were obtained from ovaries of domestic bitches (8 months old to 7 years old), and in-vitro maturation was evaluated in TCM-199 supplemented with different concentrations (0, 20, 30 or 50%) of bovine COCs-conditioned medium (BCM). The canine COCs were cultured for 72 or 96 h at 38.5°C in 5% CO₂, 5% O₂ and 90% N₂. The bovine COCs-conditioned medium was obtained from culture of bovine COCs with TCM-199 supplemented with 5% FCS for 22 h at 38.5°C in 2% CO₂, 98% air.

Results

The proportion of germinal vesicle breakdown (GVBD) after 72 h was significantly higher (P < 0.05) in medium supplemented with 30% BCM (20.7%) compared with the control group (13.4%). The rates of GVBD-MII stage were significantly higher (P < 0.05) when oocytes were matured with BCM at concentration of 30% (41.5%) compared with control (26.6%) after 72 h in-vitro culture. After 96 h in-vitro culture, the oocytes matured in medium supplemented with 30% BCM (5.5%) showed a significant increase (P < 0.05) in the proportion of MII compared with control (0.7%). However, increasing the cultivation time from 72 to 96 h resulted in an increase in oocyte degeneration rate.

Conclusions

The results suggested that bovine COCs-conditioned medium supplementation significantly increased nuclear maturation of canine oocytes.     

哺乳动物卵泡相关代谢组学研究进展

卵母细胞质量是决定辅助生殖结局的一项重要因素,创建一种简单可靠的卵母细胞质量评价体系有助于提高辅助生殖的成功率。新兴的代谢组学有望成为评估卵母细胞质量的新方法。目前,代谢组学已广泛应用于女性生殖相关的科学研究,通过对卵母细胞、颗粒细胞和卵泡液等样品中的中小分子进行定性和定量分析,研究者们寻找到多种与女性不孕相关的标志性代谢物,涉及糖类、脂质和氨基酸等多种物质的代谢,对于临床不孕症的诊断和治疗具有潜在的应用价值。综述近年哺乳动物卵泡相关的代谢组学研究。     

酸中毒时心脏克隆钾离子通道Kv1．4的动力学特性

目的：探讨心脏克隆钾离子通道Kv1．4的C型失活(Kv1．4△N)在非洲蟾蜍卵母细胞上表达后的动力学特性以及酸中毒时的改变。方法：将Kv1．4△N cRNA(最大体积为50 n1)注入非洲爪蟾的卵母细胞内,于18℃孵育16h以上。电极采用两步法拉制,微电极由1．5 mm口径的电极拉制。电极内充3M KCl,电阻0．5～1．0MΩ。采用双微电极电压钳制法(two electrode voltage clamp,TEV)在室温下(20～24℃)记录电流。结果：与正常pH时相比,酸性环境下,Kv1．4△N的峰电流减小,在pH7．4时通道在去极化至-40mv激活,而pH 6．8时为-30mv激活；通道在pH 7．4时最大失活为0．384±0．072,而在pH6．8时为0．197±0．013；在pH6．8时通道复活减慢(P＜0．05)。结论：酸中毒导致通道电流减小,并且使通道失活加快和恢复减慢。     

Fertility Preservation Methods in Breast Cancer

Thanks to the recent advances in reproductive medicine, more and more young women with breast cancer may be offered the possibility of preserving their fertility. Fertility can be endangered by chemotherapy, by treatment duration and by patient's age at diagnosis. The currently available means to preserve a young woman's fertility are pharmacological protection with gonadotrophin-releasing hormone analogues during chemotherapy, and ovarian tissue or oocyte/embryo freezing before treatment. New future venues, including in vitro maturation, will improve the feasibility and efficacy of the fertility preservation methods in breast cancer patients.     

不同操作温度对小鼠卵母细胞体外受精的影响

目的：观察不同操作温度对小鼠体外受精以及后续胚胎发育的影响,探讨较合适的实验室受精及操作温度,为提高人类体外受精-胚胎移植（IVF-ET）技术提供实验依据。方法对IVF实验室温度共设置19℃、21℃、23℃、25℃、27℃、29℃及31℃温度点,选择10~12周龄（20~25g）的ICR雌性小鼠及雄性小鼠168只。每个温度点设自然受精组和体外受精组,随机分配雌鼠8只、雄鼠4只。在上述实验室温度下对小鼠自然受精卵团与体外受精卵团进行操作、观察,并统计受精率、卵裂率、囊胚形成率及囊胚孵出率。结果体外受精组中25℃组与19℃、21℃、29℃、31℃四组间比较受精率（94.83%、52.69%、78.98%、82.76%、66.67%）、囊胚形成率（87.12%、54.17%、68.75%、70%、56.56%）、囊胚孵出率（86.62%、46.15%、66.23%、67.857%、51.78%）有统计学差异（P＜0.05）,尤以19℃、31℃差异明显;体外受精组中25℃组与19℃、21℃、31℃三组间比较卵裂率（98.79%、81.82%、90.32%、89.83%）有统计学差异（P＜0.05）,尤以19℃、31℃差异明显;顺次两组间比较,19℃与21℃、29℃与31℃组受精率、囊胚形成率、囊胚孵出率有统计学差异（P＜0.05）;组内比较19℃、21℃、29℃、31℃组各自的受精率、卵裂率、囊胚形成率、囊胚孵出率有统计学差异（P＜0.05）,尤以19℃、31℃组差异明显。结论实验室温度在23℃~27℃之间时,小鼠卵母细胞体外受精组与自然受精组在受精率、卵裂率、囊胚形成率和囊胚孵出率等方面较其他温度具有显著优势,提示23℃~27℃为较合适的实验室受精及胚胎操作温度范围。