家蚕幼虫血细胞密度变化成因及血细胞密度与耐高温性的关系

[目的]血细胞是昆虫血淋巴免疫的主导者.调查家蚕Bombyx mori幼虫血细胞密度变化和成因、血细胞密度与家蚕抗性的关系,是研究家蚕血细胞相关的免疫调控和抗性育种的重要组成.[方法]用细胞计数板统计家蚕品种大造不同龄期(4龄第1-4天、5龄第1-8天和上蔟期)幼虫10 μL血淋巴中的血细胞数目并计算血细胞密度,利用I...     

西双版纳勐海县勐阿水库对亚洲象迁移活动和人—象冲突的潜在影响

2020年1—12月,采用半结构式访查法、痕迹跟踪调查法和无人机跟踪调查法相结合的方法,对拟建西双版纳勐海县勐阿水库涉及区域的亚洲象种群结构、迁移路线等进行了调查,并采用样方法和3S技术对该区域亚洲象取食植物和栖息地适宜性进行了调查和分析,以探讨拟建勐阿水库可能对亚洲象迁移活动和人—象冲突的潜在影响。结果表明：(1)拟建勐阿水库区域活动的亚洲象小种群被称为澜沧江西部种群或西双版纳勐海—普洱澜沧种群,仅由19头亚洲象(10头雄性,9头雌性)组成,有2条亚洲象迁移路线穿越该区域;(2)在研究区域共统计到亚洲象取食植物12目19科32属33种,基本能满足该象群的取食需求;(3)海拔、坡度、植被隐蔽度及食物资源等生态因子的适宜性分析结果显示,大部分区域(包括占总面积14.61%的最适生境和占总面积82.05%的相对适宜生境)都能满足该亚洲象小种群的基本生活需求;(4)2条亚洲象迁移路线中的1条将因水库建设而阻断,迫使该象群改变原来的迁移路线;(5)水库淹没区的植被将被永久破坏,原本连片的适宜栖息地也将受到进一步切割和压缩影响;(6)当食物资源无法满足亚洲象生存需求时,它们可能会选择进入可在较短时间内获得大量食物的农耕区取食农作物和经济作物,随着人流、车流大量增加,亚洲象与人相遇的概率也会大幅度增加。分析认为,拟建勐阿水库将淹没1条迁移通道,对亚洲象迁移活动造成阻碍,迫使象群改变路线,还可能导致更为严重的人—象冲突。建议在水库工程设计和建设过程中采取有效的保护管理措施,减少工程项目对亚洲象种群及其栖息地的负面影响;水库建设和管理部门、林业和草原管理部门等应加强对亚洲象活动的监测和预警,避免亚洲象肇事造成人员伤亡和更大的经济损失。     

Demecolcine-induced enucleation of sheep meiotically maturing oocytes

The objective of this study was to investigate the possible effect of demecolcine, a microtubule-disrupting reagent, on induced enucleation (IE) of sheep meiotically maturing oocytes. Immunofluorescent staining with anti-tubulin antibodies was used to examine the spindle status of the oocytes. When the oocytes with intact germinal vesicles (GV) were cultured in the medium containing various concentrations of demecolcine (0.01 to 0.4 microg.mL-1) for 20 to 22 h, the spindle microtubule organization and first polar body (PB1) extrusion were inhibited by demecolcine in a dose-dependent manner. The highest IE rate (58.1%) was from the treatment with 0.04 microg.mL-1 demecolcine. Demecolcine treatment applied after germinal vesicle breakdown (GVBD) or at metaphase (M) yielded a PB1 extrusion rate and IE efficiency similar to the treatment applied at the onset of maturation. Analysis by immunofluorescence showed that both nonspindle microtubules and spindle microtubules were significantly disorganized by demecolcine. Combination treatment with demecolcine and cycloheximide (CHX) or 6-dimethylaminopurine (6-DMAP) led to single pronuclear formation rather than PB1 extrusion. When demecolcine-treated oocytes were transferred into demecolcine-free medium, the ability to extrude PB1 was quickly restored and a 72.1% IE rate was obtained following such treatment. These results demonstrate that demecolcine can be used as a potential reagent for induced enucleation of sheep meiotically maturing oocytes and may greatly facilitate research in nuclear transfer.     

林分因子对云顶山不同人工林林下植物多样性的影响

该研究采用典型抽样法,以成都云顶山5种人工林——柏木 枫杨林(BF)、银杏 楠木林(YN)、光皮梾木 香樟林(GZ)、枫杨 桤木林(FQ)、柏木林(CB)为研究对象,分析不同人工林的林下植物组成与多样性特征,并确定影响林下植物多样性的主导林分因子,为当地人工林经营管理提供理论依据。结果表明：(1)研究区共记录林下植物168种,隶属于62科130属;5种人工林灌木层与草本层的科属种数均以GZ最多。(2)5种不同人工林灌木层与草本层的优势种数分别为7、4、7、6、4种和5、4、9、9、10种,数量都较少。(3)5种人工林的Shannon Wiener多样性指数(H)、Simpson优势度指数(H′)、物种丰富度指数(D)、Pielou均匀度指数(J_sw) 均基本表现为草本层>灌木层,BF、GZ灌木层的D略高;灌木层的H、H′、D值均以GZ最大,但不同人工林的J_sw差异不显著;草本层的H、H′、D、J_sw均基本呈现CB>FQ>GZ>BF>YN趋势,GZ的D值略高于FQ。(4)6个林分因子对灌木层4个物种多样性指数的影响均无显著差异;林分平均树高、平均枝下高、平均胸径、平均冠幅和林分密度是影响草本层H、D的主要因子,但各林分因子对草本层H′、J_sw的影响差异不显著。研究认为,林分结构对林下草本层物种多样性的影响更大,平均树高、平均枝下高、平均胸径、平均冠幅、林分密度对草本层多样性有显著影响。     

侧金盏胚和胚乳发育的细胞胚胎学研究

利用石蜡切片技术对毛茛科植物侧金盏胚及胚乳发育进行了研究,以明确其胚胎发育的特征,为毛茛科植物的系统研究提供资料。结果表明:(1)侧金盏胚的发育属于柳叶菜型,胚乳发育为核型;初生胚乳核的分裂早于合子的第一次分裂。(2)种子成熟时,种胚尚未分化完全,尚处于球形胚后期或心形胚早期阶段,整个胚发育大约需要50~60d。(3)侧金盏种子存在明显的形态生理休眠现象,经后熟作用逐渐完成种胚的分化与生长,形成子叶形胚;侧金盏种子在相同处理条件下胚分化和发育的速度存在差异。     

Apocynin attenuates renal fibrosis via inhibition of NOXs-ROS-ERK-myofibroblast accumulation in UUO rats

Background: Oxidative stress has been identified as an important pathogenesis mechanism in the development of renal interstitial fibrosis in unilateral ureteral obstruction (UUO). Previous studies have demonstrated increased expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOXs) in response to UUO. We aimed to investigate whether NOXs activation was involved in the development of renal fibrosis in UUO by contribution to oxidative stress and the potential mechanism in the present study.

Methods: Apocynin, a NOXs inhibitor, was initiated immediately by gavage after UUO was performed on Wistar rats and continued until 7 days after UUO. Changes of markers of oxidative stress, renal macrophage infiltration and fibrosis, TGF-β1 expression, NOXs expression and activity, and ERK activation were evaluated.

Results: Apocynin significantly attenuated the activity of NOXs, accompanied with decreased expression of NOX2, NOX4, and oxidative stress markers in the obstructed kidneys of UUO. Additionally, collagen deposition and renal fibrosis induced by UUO were attenuated by apocynin treatment. Furthermore, apocynin treatment significantly attenuated the phosphorylation of ERK, accumulation of myofibroblast and infiltration of macrophage in obstructed kidneys. No significant effect of apocynin on UUO-induced increased TGF-β1 expression could be observed. And there was no significant change of anti-oxidants enzyme activities in the obstructed kidneys of apocynin-treated rats.

Conclusions: These results suggested that apocynin might exert beneficial effects on renal fibrosis by inhibition of NOXs activation and subsequent reduction of oxidative stress, ERK activation, and myofibroblast accumulation in UUO rats. Targeting NOXs may serve as a therapeutic strategy for the treatment of renal fibrosis.     

农田-荒地边缘地带中蝗虫边缘反应分析

探讨了中华蚱蜢等10种蝗总科昆虫对农田－荒地的边缘反应。研究发现,就同一边缘而言,有些物种的多度在靠近边缘时上升,而有些物种则下降。从科的水平上分析的结果与物种水平上的分析有差异。为了更精确地了解边缘对蝗虫的分布格局的影响,从物种水平上对其进行分析是 很有必要的。     

Therapeutic inhibition of hepatitis B virus surface antigen expression by RNA interference

RNA interference (RNAi) mediated inhibition of virus-specific genes has emerged as a potential therapeutic strategy against virus induced diseases. Human hepatitis B virus (HBV) surface antigen (HBsAg) has proven to be a significant risk factor in HBV induced liver diseases, and an increasing number of mutations in HBsAg are known to enhance the difficulty in therapeutic interventions. The key challenge for achieving effective gene silencing in particular for the purpose of the therapeutics is primarily based on the effectiveness and specificity of the RNAi targeting sequence. To explore the therapeutic potential of RNAi on HBV induced diseases in particular resulted from aberrant or persistent expression of HBsAg, we have especially screened and identified the most potent and specific RNAi targeting sequence that directly mediated inhibition of the HBsAg expression. Using an effective DNA vector-based shRNA expression system, we have screened 10 RNAi targeting sequences (HBsAg-1 to 10) that were chosen from HBsAg coding region, in particular the major S region, and have identified four targeting sequences that could mediate sequence specific inhibition of the HBsAg expression. Among these four shRNAs, an extremely potent and highly sequence specific HBsAg-3 shRNA was found to inhibit HBsAg expression in mouse HBV model. The inhibition was not only preventive in cotransfection experiments, but also had therapeutic effect as assessed by post-treatment protocols. Moreover, this HBsAg-3 shRNA also exhibited a great potency of inhibition in transgenic mice that constitutively expressed HBsAg. These results indicate that HBsAg-3 shRNA can be considered as a powerful therapeutic agent on HBsAg induced diseases.     

Novel knowledge-based mean force potential at the profile level

Background  

The development and testing of functions for the modeling of protein energetics is an important part of current research aimed at understanding protein structure and function. Knowledge-based mean force potentials are derived from statistical analyses of interacting groups in experimentally determined protein structures. Current knowledge-based mean force potentials are developed at the atom or amino acid level. The evolutionary information contained in the profiles is not investigated. Based on these observations, a class of novel knowledge-based mean force potentials at the profile level has been presented, which uses the evolutionary information of profiles for developing more powerful statistical potentials.     

真核表达载体pcDNA3.1(-)+KG的构建及在HeLa细胞中的表达

为了将绿色荧光蛋白(green fluorescent protein,GFP)引入细胞核内,采用两轮PCR方法从原先克隆在pcD-NA3.1(-)+GFP载体中将GFP编码序列扩增出来并引入Kozak序列和核定位信号,使用常规酶切和连接方法将其重组至pUCm-T克隆载体中,再将目的片段重组至pcDNA3.1(-)中,对阳性克隆进行酶切、PCR和测序鉴定后,构建了带有Kozak序列和核定位信号的绿色荧光蛋白(GFP)真核表达载体pcDNA3.1(-)+KG。真核表达载体pcDNA3.1(-)+KG被转染试剂Su-perfect转染至HeLa细胞中,绿色荧光蛋白基因在HeLa细胞中得到表达而且在细胞核中观察到绿色荧光。该研究以绿色荧光蛋白为标记初步建立了活体观察真核细胞核动态变化的研究体系。