一种适合杨树RNA提取的方法

本试验为三种杨树材料建立了一种杨树RNA提取的方法。本方法具有充分抑制材料中的多酚褐化,提高RNA产率;最大限度地去除多糖及次生代谢物质,避免与RNA共沉淀;去除蛋白质和DNA污染;提取时间短,质量高等特点。通过本方法提取的RNA OD260/OD280值在1.9~2.0之间,能够完全满足后续实验的要求,如体外翻译、cDNA文库构建、基因芯片和DDRT-PCR等。     

Preliminary evaluation of Community-Led Total Sanitation for the control of Taenia solium cysticercosis in Katete District of Zambia

Taenia solium taeniasis/cysticercosis is a zoonotic disease endemic in sub-Saharan Africa. It is associated with poor sanitary practices, free-range pig husbandry and lack of disease awareness in endemic communities. A comparative research was conducted with pre and post-intervention assessments in nine villages to evaluate Community-Led Total Sanitation (CLTS) as an intervention measure for the control of porcine cysticercosis in Katete District in the Eastern Province of Zambia. Blood samples were collected from pigs for circulating antigen detection and a questionnaire focused on the household was administered to a total of 153 respondents whose pigs were examined (64 pre-intervention, 89 post-intervention), in order to obtain information on general demographic characteristics, pig husbandry practices, sanitation practices and associated knowledge and awareness of T. solium infections. The first sampling was conducted prior to the implementation of the CLTS and second sampling eight months after triggering of CLTS in the selected villages. A total of 379 pig serum samples were examined using the B158/B60 Ag-ELISA to detect T. solium cysticercosis, 104 pre-intervention and 275 post-intervention, of which 14 (13.5%) and 45 (16.4%) were positive, respectively. Wald test p-values were computed to assess significant differences in the variables of interest mentioned above for the pre and post CLTS. The research revealed that CLTS as a control measure did not significantly improve T. solium infections in pigs. The research also revealed that the sanitation practices and awareness of cysticercosis did not change. It is recommended that a longer term evaluation be undertaken when the villages have been declared open defaecation free. In addition, the research recommends that health education, mass drug treatment and pig vaccination be incorporated, as an essential component of prevention and control programmes for T. solium infections.     

抗流感病毒药物研究进展

季节性和爆发性流感引起了高发病率和死亡率,不仅严重危害人体健康,还影响社会稳定和经济发展。目前对抗流感的主要措施是预防接种,然而流感病毒变异速度非常快,使得现有的疫苗对新变异的流感病毒无效或者预防效果不好。因此,抗病毒药物的研究显得非常必要,而被深入研究的流感病毒的生命周期和病毒结构蛋白也推进了抗流感病毒药物设计的进展。本文将对抗流感药物设计靶向进行概述,旨在为今后研究开发出更高效、更理想的多靶向抗流感病毒药物提供借鉴。     

单细胞测序对绒山羊毛乳头细胞的鉴定

【目的】 基于单细胞测序技术探究绒山羊毛乳头细胞标记基因,优化毛乳头细胞体外鉴定的方法,为研究绒山羊毛囊发生发育提供良好的细胞模型。【方法】 运用Seurat单细胞分析法分析陕北白绒山羊胚胎期60、90、120 d皮肤组织的单细胞测序数据。原始数据经质控、过滤、标准化后,采用UMAP方法进行数据降维与聚类分析。通过对比已报道的毛囊相关标记基因,获得完整绒山羊毛囊细胞类群谱系信息。通过基因差异表达分析,筛选绒山羊毛乳头细胞特异性标记基因。利用免疫荧光试验鉴定标记蛋白在绒山羊皮肤组织中的表达与分布情况,进一步筛选毛乳头区域特异性表达蛋白。体视镜下机械分离完整绒山羊毛囊,采用酶消化法分离绒山羊毛乳头区域并在体外培养至细胞分离,最终通过差速贴壁法纯化细胞。待毛乳头细胞纯度较高时,利用免疫荧光试验验证候选标记蛋白在分离细胞中的表达情况。【结果】 从单细胞水平分析了绒山羊毛囊发生过程中涉及的关键细胞转录信息,成功获得绒山羊皮肤中的17个主要细胞类群信息,鉴定出包括真皮细胞谱系、表皮细胞谱系、毛乳头细胞、毛干细胞、内根鞘细胞等绒山羊皮肤结构关键细胞类群,以及周皮细胞、巨噬细胞、肌肉细胞等其他功能细胞类群。筛选获得毛乳头细胞特异性基因427个,包括SOX2、FGF7、APOD、BMP3、HHIP、HEY2和SPON1等,这些基因在绒山羊毛乳头细胞中的表达丰度远高于其他细胞类型,被认为是毛乳头细胞特异性标记基因。组织免疫荧光试验进一步证明SOX2、FGF7与APOD等蛋白在绒山羊毛乳头区域内特异性表达,可用于皮肤组织中绒山羊毛乳头细胞的定位。此外,本研究在成功分离单根绒山羊次级毛囊的基础上,实现了绒山羊毛乳头区域的贴壁培养,成功观测到大量细胞从毛乳头区域逐步迁移分离的过程。细胞免疫荧光试验结果显示SOX2、FGF7与APOD均在绒山羊毛乳头细胞中表达,且SOX2阳性细胞约占毛乳头细胞总量的76%左右,而FGF7和APOD阳性细胞则占98%以上。结合绒山羊皮肤组织免疫荧光定位结果,SOX2、FGF7与APOD等标记可用于鉴定体外分离培养的绒山羊毛乳头细胞。【结论】 利用单细胞测序技术描述了绒山羊主要皮肤细胞的转录组信息,筛选出毛乳头细胞特异性表达基因。经免疫荧光试验证实,单细胞测序鉴定标记基因的方法简单高效,且具备较高准确率。筛选的SOX2、FGF7与APOD不仅为绒山羊毛乳头细胞体内定位提供了有效的标记,而且为多标记鉴定毛乳头细胞提供可能,更为进一步研究毛囊发育相关基因的功能及调控机制奠定重要基础。     

Diagnostic evaluation and short-term outcome as indicators of long-term prognosis in horses with findings suggestive of inflammatory bowel disease treated with corticosteroids and anthelmintics

Background

Recurrent colic and unexplained weight loss despite good appetite and adequate feeding and management practices are common conditions in the horse. However, little information has been published on the systematic diagnostic evaluation, response to treatment, prognostic factors or outcome of either presentation. The aims of this study were to 1) identify possible prognostic indicators and 2) report the short- and long-term response to treatment with corticosteroid therapy of a variety of horses with a presumptive diagnosis of inflammatory bowel disease (IBD).Thirty-six horses with a history of recurrent colic and/or unexplained weight loss were screened with a detailed clinical, clinicopathological and diagnostic imaging examination. Twenty horses were subsequently selected that had findings consistent with inflammatory bowel disease based on the fulfilment of one or more of the following additional inclusion criteria: hypoproteinaemia, hypoalbuminaemia, malabsorption, an increased intestinal wall thickness on ultrasonographic examination or histopathological changes in rectal biopsy. These 20 horses were treated with a standardized larvicidal anthelmintic regime and a minimum of three weeks of corticosteroid therapy.

Results

The initial response to treatment was good in 75% (15/20) of horses, with a 3-year survival rate of 65% (13/20). The overall 3-year survival in horses that responded to initial treatment (12/15) was significantly higher (P = 0.031) than in those that did not respond to initial treatment (1/5). The peak xylose concentration was significantly (P = 0.048) higher in survivors (1.36 ± 0.44 mmol/L) than non-survivors (0.94 ± 0.36 mmol/L).

Conclusions

The overall prognosis for long-term survival in horses with a presumptive diagnosis of IBD appears to be fair to moderate, and the initial response to anthelmintic and corticosteroid therapy could be a useful prognostic indicator. The findings of the present study suggest that a low peak xylose concentration in absorption testing is associated with a less favourable prognosis, supporting the use of this test.     

布鲁菌感染小鼠RAW264．7细胞对IRGl基因的表达调控作用

针对小鼠RAW264．7细胞IRGl基N设计4个RNA干扰靶位,筛选出最佳干扰序列构建shRNA慢病毒载体质粒并包装获得慢病毒颗粒,进而经嘌呤霉素筛选获得稳转细胞系,实现IRGl基因在RAW264．7细胞基因表达的沉默。并通过布鲁菌l6M株及M5株感染基因沉默细胞对IRGl基因在布鲁菌感染中的作用进行研究。结果表明,慢病毒介导的shRNA高效、稳定地沉默了IRGl基因的表达,布鲁菌侵染RAW264．7细胞后IRGl基因表达上调。未试验为1RG1基因及相美调控基因抗布鲁菌病作用研究奠定了基础。     

基于全站仪精准量测树木材积方法初探

笔者根据北京林业大学冯仲科教授提出的精准林业的理念,使用全站仪对树木材积进行量测,并将全站仪量测的材积数据与解析法所得砍伐数据进行比较,结果表明,全站仪量测立木材积的精度在91%以上,平均精度在96%左右,完全能满足高精度量测立木材积的要求,建议推广应用。     

树皮内含物对香梨优斑螟寄主选择的影响

为了明确香梨优斑螟的寄主选择和寄主树皮内含物对寄主选择的影响,调查研究野外香梨优斑螟在不同寄主植物上的危害情况和室内幼虫的取食选择,并对寄主植物树皮中几种内含物含量与香梨优斑螟的寄主选择关系进行分析。结果表明:在野外,不同寄主植物的受害率、平均虫口数及最大虫口数,均为香梨树苹果树枣树,但枣树的蛹与幼虫的比值高于苹果;室内幼虫取食选择试验,幼虫对4种寄主植物的选择存在显著差异,香梨树枣树砀山梨树苹果树,推测枣树为香梨优斑螟潜在的高风险寄主。香梨优斑螟幼虫的寄主选择与单一的可溶性糖、可溶性蛋白、总糖、氨基酸和单宁含量均无显著相关,与4种树的碳氮比(r=-0.960,P0.05)和总酚含量(r=-0.900,P0.05)呈显著负相关,均衡的营养和较低的取食抑制物含量是香梨优斑螟选择寄主植物的标准。