Preliminary experiments on mechanical stretch-induced activation of skeletal muscle satellite cells in vivo

We have shown in vitro that mechanical stretch triggers activation of quiescent satellite cells of skeletal muscle to enter the cell cycle through an intracellular cascade of events including nitric oxide (NO) synthesis that results in the release of hepatocyte growth factor (HGF) from its extracellular association and its subsequent presentation to signaling receptors. In order to explore the activation mechanism in vivo, stretch experiments were conducted in the living animal using our suspension model developed. This system used the weight of the hind portion of rats to stretch the inside muscles of the left hind limb suspended for a period of 0.5–2.0 h. At the end of the stretch period, the rats received an intraperitoneal injection of bromodeoxyuridine followed by immunocytochemistry for its incorporation as an index of satellite cell activation in vivo. Depending on the period of stretch, bromodeoxyuridine labeling was increased significantly over the contralateral unstretched leg or control muscle from untreated rats. A stretched muscle extract prepared from the 2 h stretched tissue by incubating it in PBS, showed the active form of HGF as revealed by immunoblotting and it could stimulate the activation of unstretched satellite cells. Also, administering NO synthase inhibitor L‐NAME prior to muscle stretch abolished the stretch activation of satellite cells. Therefore, the results from these experiments demonstrate that stretching muscle triggers NO synthesis and HGF release, which could activate satellite cells in vivo.     

卵母细胞激活的信号机制

本文主要阐述精子与卵母细胞质膜相互作用的信号转变,特别是精子启动这些信号途径的机制,并讨论与精子诱导的Ca2 振荡发生反应并在卵母细胞激活中起重要作用的下游信号分子。     

序贯培养法对猪孤雌激活胚胎发育能力的影响

经体外成熟、孤雌激活和培养获得猪胚胎,研究了不同培养体系、共培养体细胞和序贯培养对猪孤雌激活胚胎发育的影响。试验表明:孤雌激活卵母细胞在SOF 10?S培养体系中分裂效果最好,添加胎牛血清的NCSU-23和颗粒细胞对胚胎发育有促进作用,培养6d后发育到桑囊胚的比率增加(P<0.05)。在序贯培养的前3d,SOF培养基(不含葡萄糖)和颗粒细胞对胚胎的发育有促进作用,分裂率(P<0.05)和突破4细胞阻滞的数目显著增加,在培养的后3d,添加胎牛血清的NCSU-23和输卵管上皮细胞能支持较多胚胎发育到桑囊胚,桑囊胚的发育率为(59.5±3.2)%(P<0.05)。结果表明,SOF培养基和颗粒细胞 添加胎牛血清的NCSU-23和输卵管上皮细胞的序贯培养系统能较好的促进胚胎的发育。     

Physical activation of biochar and its meaning for soil fertility and nutrient leaching – a greenhouse experiment

The slow alteration of the surface of charred biomass (biochar) over time may contribute to an improved nutrient retention and thus fertility of tropical soils. Here, we investigated soils from temperate climates and investigated whether a technical steam activation of biochar could accelerate its positive effects on nutrient retention and uptake by plants relative to nonactivated biochar. To this aim, we performed microcosm experiments with sandy or silty soil, mixed with 2.0, 7.5 and 15.0 g/kg soil of fine (<2 mm) or coarse‐sized (2–10 mm) biochar from beech wood (Fagus sp.). After initial fertilizer (NPK), ashes and excess nutrients were leached with water, and the microcosms were planted for 142 days with Italian Ryegrass (Lolium multiflorum ssp. italicum). Thereafter, leachate, soil and plant samples were analysed for their nutrient contents. The results showed that biochar additions of ≤15 g/kg soil left elevated contents of available P and N in the surface soil but reduced their uptake into the plants. As a result, total biomass production was unchanged. Different particle size and application amounts influenced these findings only marginally. Nitrate leaching was enhanced in the sandy soil (+41% for nitrate, but reduced in the silty soil ?17%) and P was immobilized. Hence, the fertility of the temperate soils under study was only marginally affected by pure biochar amendments. Steam activation, however, almost doubled the positive effects of biochars in all instances, thus being an interesting option for future biochar applications.     

铅酸蓄电池复合脉冲活化原理及电路设计

分析铅酸蓄电池充、放电化学反应过程及其硫化现象、原因和危害,探讨解决硫化问题的途径和主要方法,研究铅酸蓄电池脉冲充电的活化原理,设计了一种复合脉冲循环充放电工作电路.为铅酸蓄电池活化与修复研究提出了新的观点和应用技术.     

桑蚕卵电晕人工孵化技术研究：V.电晕处理蚕种的农村饲育

通过１９９４～１９９６年电晕处理蚕种的农村饲养，得到以下几点认识，在保证卵面消毒和排除孵化不齐因素情况下，电晕蚕种与浸酸蚕种的产量质量无显著差异，专供农用的大板电晕仪存在放电不匀，孵经不齐和孵化率欠高的问题，有待改进，电晕处理蚕种还须进行卵消毒。     

马尾松种源对磷肥的遗传反应及根际土壤营养差异

利用5a生马尾松种源与磷肥互作试验林，研究不同种源对磷肥的遗传反应式样以及在低(缺)磷环境下的根际营养差异。结果发现，广东高州和广西岑溪两种源对磷肥的反应显著，属于对磷肥敏感型种源。广东信宜和福建武平两种源对磷肥反应不敏感，属于耐低磷型或对磷肥不敏感型种源，而江西崇义种源显著的磷肥效应仅在造林后头2～3年显示。对于根际土的的化学性质，不同种源虽有较大差异，但根际土的有机质、全氮、水解氮和有效磷含量一般显著高于非根际土。不同种源根际对土壤中的磷都具有活化作用和富集作用，尤其是广东信宜和福建武平两种源对土壤磷的活化和富集作用最为强烈，这可初步解释这两个种源对磷肥不敏感、能适应低(缺)磷胁迫的遗传机制。马尾松不同种源根际土壤并未发生明显的酸化。     

稳定表达山羊SLAM受体的BHK-21细胞系的建立及应用

信号淋巴激活分子(SLAM)又称为CD150,是小反刍兽疫病毒(PPRV)和犬瘟热病毒(CDV)等麻疹病毒属病毒感染淋巴细胞的主要受体,在病毒侵入细胞中发挥重要作用。为建立稳定表达山羊SLAM(g SLAM)的真核细胞系,本研究将人工合成的g SLAM基因克隆至真核表达质粒p IRESpuro3中,并在该基因的3'端引入Flag标签序列作为分子标记,构建了重组质粒p IRES3-g SLAM。将该重组质粒转染BHK-21细胞,经嘌呤霉素加压筛选及采用表达绿色荧光蛋白(GFP)的重组PPRV病毒(r PPRV/GFP)感染鉴定后,筛选到稳定表达g SLAM基因的细胞系。r PPRV/GFP感染和western blot鉴定表明,无论是否有嘌呤霉素压力的存在,该细胞系在传代至第20代,仍能稳定表达g SLAM蛋白。由于g SLAM氨基酸序列与犬的同源性较高,以表达GFP重组CDV强毒株(r CDV/GFP)感染该细胞系,病毒可以感染且能形成明显的细胞病变,表明该细胞系可用于CDV强毒分离和致弱机制等相关研究。     

No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse

Background

This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation.

Results

First, mouse metaphase II (MII) oocytes were vitrified in the open-pulled straws (OPS). After warming, they were cultured for 1 h in M₂ medium containing melatonin at different concentrations (0, 10⁻⁹, 10⁻⁷, 10⁻⁵, 10⁻³ mol/L). Then the oocytes were used to detect reactive oxygen species (ROS) and glutathione (GSH) levels (fluorescence microscopy), and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10⁻³ mol/L melatonin group was significantly lower than that in melatonin-free group (control). The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp90aa1, Hsf1, Hspa1b, Nrf2 and Bcl-x1 with qRT-PCR in oocytes treated with 10⁻⁷, or 10⁻³ mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90aa1 expression in 10⁻⁷ mol/L melatonin-treated group than in the control (P < 0.05); the Hsf1, Hsp90aa1 and Bcl-x1 expression were significantly decreased in 10⁻³ mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10⁻⁷ or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them.

Conclusions

Our results indicate that the supplementation of melatonin (10⁻⁹ to 10⁻³ mol/L) in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.