A simplified method for the reconstruction of fully competent mouse zygotes from adult somatic donor nuclei

Until now, full-term development of mouse embryos reconstructed from somatic nuclei has been convincingly achieved only when a piezoimpact pipette drive unit is used for the injection of a donor nucleus into an enucleated recipient oocyte. Here we describe a simplified method for mouse cloning that requires neither electrofusion nor a piezo device. Efficient rates of enucleation can be achieved without staining the chromosomes of the recipient oocyte and high survival rates are obtained after direct injection of the donor nucleus. Although a low proportion of reconstructed embryos could implant after their transfer into the oviducts of foster mothers (less than 5%), we show that at least some of them can develop into normal young.     

Effect of gender on the development of hypocapnic apnea/hypopnea during NREM sleep.

We hypothesized that a decreased susceptibility to the development of hypocapnic central apnea during non-rapid eye movement (NREM) sleep in women compared with men could be an explanation for the gender difference in the sleep apnea/hypopnea syndrome. We studied eight men (age 25-35 yr) and eight women in the midluteal phase of the menstrual cycle (age 21-43 yr); we repeated studies in six women during the midfollicular phase. Hypocapnia was induced via nasal mechanical ventilation for 3 min, with respiratory frequency matched to eupneic frequency. Tidal volume (VT) was increased between 110 and 200% of eupneic control. Cessation of mechanical ventilation resulted in hypocapnic central apnea or hypopnea, depending on the magnitude of hypocapnia. Nadir minute ventilation in the recovery period was plotted against the change in end-tidal PCO(2) (PET(CO(2))) per trial; minute ventilation was given a value of 0 during central apnea. The apneic threshold was defined as the x-intercept of the linear regression line. In women, induction of a central apnea required an increase in VT to 155 +/- 29% (mean +/- SD) and a reduction of PET(CO(2)) by -4.72 +/- 0.57 Torr. In men, induction of a central apnea required an increase in VT to 142 +/- 13% and a reduction of PET(CO(2)) by -3.54 +/- 0.31 Torr (P = 0.002). There was no difference in the apneic threshold between the follicular and the luteal phase in women. Premenopausal women are less susceptible to hypocapnic disfacilitation during NREM sleep than men. This effect was not explained by progesterone. Preservation of ventilatory motor output during hypocapnia may explain the gender difference in sleep apnea.     

Recording of calcium transient and analysis of calcium removal mechanisms in cardiac myocytes from rats and ground squirrels

With confocal microscopy, we recorded calcium transients and analyzed calcium removal rate at different temperatures in cardiac myocytes from the rat, a non-hibernator, and the ground squirrel, a hibernator. The results showed a remarkable increase of the diastolic level of calcium transients in the rat but no detectable change in the ground squirrel. Calcium transient of the ground squirrel, compared with that of the rat at the same temperature, had a shorter duration and showed a faster calcium removal. As indicated by the pharmacological effect of cyclopiazonic acid, calcium uptake by sarcoplasmic reticulum (SR) was the major mechanism of calcium removal, and was faster in the ground squirrel than in the rat. Our results confirmed the essential role of SR in hypothermia-tolerant adaptation, and negated the importance of Na-Ca exchange. We postulated the possibility to improve hypothermia-tolerance of the cardiac tissue of non-hibernating mammals.     

丙型肝炎病毒RNA多聚酶在昆虫细胞中的表达

ＨＣＶ ＮＳ５Ｂ基因片段克隆入ＢＡＣ－ＴＯ－ＢＡＣ＾ＴＭ重组杆状病毒表达系统的ｐＦＡＳＴＨＴｃ载体质粒,转化ＤＨ１０ＢＡＣ＾ＴＭ感受态细菌获得重组的Ｂａｃｍｉｄ质粒,将重组Ｂａｃｍｉｄ质粒转染Ｓｆ细胞,获得的重组杆状病毒可表达目的蛋白。免疫印迹和体外活性检测表明,所表达蛋白为ＨＣＶ ＮＳ５Ｂ蛋白,具有多聚酶活性。     

虎杖根茎蒽醌类化合物细胞化学定位和含量测定(简报)

虎杖(Polygonum cuspidatum Sidb.et Zucc.)为蓼科多年生灌木状草本。其根茎入药。主要化学成分有蒽醌、萘醌、酚类和多糖化合物;其中蒽醌类化合物是虎杖的主要药用有效成分,包括大黄素、大黄酚,大黄素甲醚和大黄酸等。虎杖的蒽醌类化合物具显著的抗病毒、抗菌和消炎作用;并可保护心肌,防止血栓形成,改善微循环,提高细胞免疫力,也是致癌基因转导信号的抑制因子。关于虎杖蒽醌类化合物细胞化学研究尚未见报道。刘文哲等曾对贯叶连翘分泌结构中金丝桃素和大黄中蒽醌类同化合物进行了组织化学研究。本文对不同生长期虎杖的根茎以及同     

云南干热河谷地区种子植物资源与开发利用初探

云南干热河谷植被均为耐旱的旱生植被,主要有以下三种类型：1）半稀树草原（Semi-savanna）型植被;2）肉质多刺灌丛;3）在金沙江上游滇西北的亚热带性的干热河谷地区分布着类似马基(Maquis-like）型的多刺小叶矮生灌丛。按其产物和用途可分为6类：药用植物、纤维植物、树脂和树胶植物、芳香油植物以及淀粉植物等。资源破坏性开发和环境的变化,使现存资源的保护和开发利用成为目前的主要问题。     

Generation of dwarf goat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes

The developmental potential of caprine fetal fibroblast nuclei after in vitro transfection and nuclear transfer (NT) into enucleated, in vitro-matured oocytes was evaluated. Fetal fibroblasts were isolated from Day 27 to Day 30 fetuses from a dwarf breed of goat (BELE: breed early lactate early). Cells were transfected with constructs containing the enhanced green fluorescent protein (eGFP) and neomycin resistance genes and were selected with G418. Three eGFP lines and one nontransfected line were used as donor cells in NT. Donor cells were cultured in Dulbecco minimum Eagle medium plus 0.5% fetal calf serum for 4-8 days prior to use in NT. Immature oocytes were recovered by laparoscopic ovum pick-up and matured for 24 h prior to enucleation and NT. Reconstructed embryos were transferred as cleaved embryos into synchronized recipients. A total of 27 embryos derived from transgenic cells and 70 embryos derived from nontransgenic cells were transferred into 13 recipients. Five recipients (38%) were confirmed pregnant at Day 35 by ultrasound. Of these, four recipients delivered five male kids (7.1% of embryos transferred) derived from the nontransfected line. One recipient delivered a female kid derived from an eGFP line (7.7% of embryos transferred for that cell line). Presence of the eGFP transgene was confirmed by polymerase chain reaction, Southern blotting, and fluorescent in situ hybridization analyses. Nuclear transfer derivation from the donor cells was confirmed by single-strand confirmation polymorphism analysis. These results demonstrate that both in vitro-transfected and nontransfected caprine fetal fibroblasts can direct full-term development following NT.     

秀丽白虾精子发生的研究

利用透射电镜观察秀丽白虾的精子发生,并根据染色质及细胞形态的变化将精子发生的全过程划分为五个时期,即精原细胞、初级精母细胞、次级精母细胞、精细胞和精子。在精子发生过程中,细胞器经历了由少到多,到最后解体特化的过程。晚期精细胞中出现单个中心粒,但在成熟精子中消失。棘突由片层复合体衍生物汇集并延伸而成。染色质在精原细胞中为部分异固缩,在精母细胞中高度凝聚为染色体,在精细胞及精子中为均匀非致密态。减数分裂同步率较高。成熟精子中帽状体和棘突构成顶体复合体。     

海岛退化生态系统的恢复

海岛在干扰极下极易退化且不易恢复,这些干扰包括毁林、引种不当和自然灾害三类。海岛恢复的限制性因子是缺乏淡水和土壤,生物资源缺乏、严重的风害或暴雨。不同大小的海岛和海岛不同部分的恢复策略不同。海岛植被恢复可参考其群落演替过程,其恢复至少是一个群落或生态系统水平的恢复。海岛恢复的长期利益包括重建海岛的生物群落,再现海岛生态系统的营养循环,恢复海岛的进化过程。海岛恢复的过程比较复杂,最关键的是要选择好适生的关键种。