Identification of Rabbit Oviductal Fluid Proteins Involved in Pre‐Fertilization Processes by Quantitative Proteomics

Oviductal fluid (ODF) proteins modulate and support reproductive processes in the oviduct. In the present study, proteins involved in the biological events that precede fertilization have been identified in the rabbit ODF proteome, isolated from the ampulla and isthmus of the oviduct at different time points within 8 h after intrauterine insemination. A workflow is used that integrates lectin affinity capture with stable‐isotope dimethyl labeling prior to nanoLC‐MS/MS analysis. In total, over 400 ODF proteins, including 214 lectin enriched glycoproteins, are identified and quantified. Selected data are validated by Western blot analysis. Spatiotemporal alterations in the abundance of ODF proteins in response to insemination are detected by global analysis. A subset of 63 potentially biologically relevant ODF proteins is identified, including extracellular matrix components, chaperones, oxidoreductases, and immunity proteins. Functional enrichment analysis reveals an altered peptidase regulator activity upon insemination. In addition to protein identification and abundance changes, N‐glycopeptide analysis further identifies 281 glycosites on 199 proteins. Taken together, these results show, for the first time, the evolving oviductal milieu early upon insemination. The identified proteins are likely those that modulate in vitro processes, including spermatozoa function.     

Identification of Prolificacy‐Related Differentially Expressed Proteins from Sheep (Ovis aries) Hypothalamus by Comparative Proteomics

Reproduction, as a physiologically complex process, can significantly affect the development of the sheep industry. However, a lack of overall understanding to sheep fecundity has long blocked the progress in sheep breeding and husbandry. In the present study, the aim is to identify differentially expressed proteins (DEPs) from hypothalamus in sheep without FecB mutation in two comparison groups: polytocous (PF) versus monotocous (MF) sheep at follicular phase and polytocous (PL) versus monotocous (ML) sheep at luteal phase. Totally 5058 proteins are identified in sheep hypothalamus, where 22 in PF versus MF, and 39 proteins in PL versus ML are differentially expressed, respectively. A functional analysis is then conducted including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis to reveal the potential roles of these DEPs. The proteins ENSOARP00000020097, ENSOARP00000006714, growth hormone (GH), histone deacetylase 4 (HDAC4), and 5′‐3′ exoribonuclease 2 (XRN2) in PF versus MF, and bcl‐2‐associated athanogene 4 (BAG4), insulin‐like growth factor‐1 receptor (IGF1R), hydroxysteroid 11‐beta dehydrogenase 1 (HSD11B1), and transthyretin (TTR) in PL versus ML appear to modulate reproduction, presumably by influencing the activities of gonadotropin‐releasing hormone (GnRH). This study provides an alternative method to identify DEPs associated with sheep prolificacy from the hypothalamus. The mass spectrometry data are available via ProteomeXchange with identifier PXD013822.     

脆木耳生物学特性及驯化栽培

对采自海南省白沙县鹦哥岭阔叶树腐木上的标本,根据形态学特征和多基因分析,鉴定为脆木耳Auricularia fibrillifera,经分离纯化获得纯菌株作为实验材料,为了充分利用和开发木耳属资源,首次对该种的生物学特性和驯化栽培进行了研究。研究不同碳源、氮源、无机盐和生长因子在固体培养条件下对脆木耳菌丝生长的影响,对以上4个因素进行单因素试验,从中选出3个最优的水平进行正交试验。结果表明：脆木耳菌丝最适生长温度为33℃,最佳碳源为麦芽糖,最佳氮源为牛肉粉,最佳无机盐添加量为1.5% PO₄^3--1% Mg²⁺,最佳生长因子为玉米汁。驯化栽培过程中,栽培配方为：木屑78%,麸皮20%,石膏粉1%,石灰粉1%,发菌温度为22℃可使脆木耳菌丝在40d左右满袋,且生长旺盛,可培育出子实体。     

黄精褐斑病的病原生物学特性

为明确发生在贵州省施秉县的黄精叶斑类病害的病原菌,通过形态学和分子生物学方法对病原菌进行了鉴定,并对其生物学特性进行了初步研究。形态特征及rDNA-ITS、β-tubulin和tef1多基因序列分析表明,该病原菌为棕榈拟盘多毛孢Pestalotiopsis trachicarpicola。生物学特性研究结果表明,该菌菌丝体适宜生长温度为15-30℃,最适温度为28℃;最适pH值为5;以葡萄糖为碳源、酵母浸膏为氮源比较适合菌丝体的生长;菌丝体生长的最佳培养基为PDA;光照对菌丝体生长无明显影响;菌丝体致死温度为45℃。     

灵芝蛛网病病原菌及其生物学特性

为明确吉林省蛟河市灵芝Ganoderma lingzhi栽培主产区发生的疑似灵芝蛛网病的病原菌,作者通过罹病灵芝子实体病原物的分离纯化、致病性测定、形态学和分子生物学鉴定以及病原菌的生物学特性研究,证明引起吉林省蛟河市灵芝蛛网病的病原菌为嗜菌枝葡霉Cladobotryum mycophilum。该菌营养体最适生长条件为温度25℃、pH 5、蔗糖作碳源、酵母浸粉作氮源,光照对菌丝体生长有一定的抑制作用,完全黑暗最适宜生长。本文研究结果为进一步研究该病害的发生规律和防治措施提供了理论参考。     

流域视野下梯田聚落的人居环境空间特征——以元江流域为例

针对村落人居环境研究过于注重村落本体的现状,以GIS的技术方法结合实地调研,选取梯田聚落所处的元江流域及典型样本大瓦遮河子流域为研究范围,探讨流域视野下人居环境空间的宏观与微观特征。结论如下：1）哀牢山区的崎岖山脉与密集水系造就了元江流域的格局,以水平、垂直特征奠定子流域尺度中梯田聚落的人居环境基础;2）大瓦遮河子流域的人居环境以“典型山地本底、帚状水系关联”为共性,在中低山区形成“面状森林核心、村落散布周边、旱地梯田混合”的彝族聚落区,在中山区形成“面状梯田核心、村落环绕周边、带状森林围合”的哈尼族聚落区,二者差异源自不同民族适应垂直分异效应的差异化农作策略;3）流域是人居环境研究的基本单元,超越村落本体之外的流域视野对复杂地理环境中的人居环境研究具有重要价值与普遍意义。     

结核分枝杆菌硫醇乙酰基转移酶基因敲除株的构建及其生物学特性分析

为探索硫醇乙酰基转移酶(mycothiol acetyltransferase,MshD)在结核分枝杆菌中的生物学特性,本实验利用噬菌体为载体的同源重组技术,构建结核分枝杆菌mshD基因敲除株、mshD基因回补株,用实时定量聚合酶链反应(real time-quantitative polymerase chain reaction, RT-qPCR)对所构建的菌株进行验证。分别收集H37Ra野生株、mshD基因敲除株、mshD基因回补株对数生长期菌液各5 mL, 离心收集菌体并培养,以观察菌落形态、生物膜形成及生长曲线测定;用5 mmol/L H₂O₂、0.05% SDS,50 ℃热激及低氧条件下分别处理基因敲出菌株和野生菌株,将菌液进行10倍梯度稀释,培养4～6周后检测抗胁迫能力并计算存活率。结果显示, 与野生株H37Ra相比,mshD基因敲除株菌落褶皱减少且菌落偏小,生长趋势较为缓慢;生物膜形成所需时间增长且褶皱明显减少;抗逆能力下降,存活率略低于野生株和回补株。揭示了mshD基因对结核分枝杆菌的生长具有重要作用,为进一步揭示该基因的功能和作用机制奠定了基础。     

胆囊结石患者结石形态学特征及血浆脂多糖水平与急性胆源性胰腺炎的关系研究

摘要 目的：探讨胆囊结石患者结石形态学特征及血浆脂多糖（LPS）水平与急性胆源性胰腺炎（ABP）的关系。方法：选取2015年10月~2018年9月期间武汉大学人民医院收治的胆囊结石患者164例为研究对象,分析结石形态学特征与并发ABP的关系,同时采用Logistic回归分析ABP发生的危险因素。将所有患者根据LPS水平分为低LPS组（n=65,<10 pg/mL）以及高LPS组（n=99,≥10 pg/mL）,分析血浆LPS水平对不同结石大小、不同总胆固醇（TG）水平患者并发ABP的影响。结果：多发胆囊结石、球状结石、<3 mm结石、软碎型结石患者并发ABP的概率明显高于单发胆囊结石、不规则状结石或泥沙状结石、3~10 mm结石或>10 mm结石、硬型结石或胶冻型结石（P<0.05）。Logistic回归分析结果显示,多发胆囊结石、球状结石、<3 mm结石以及软碎型结石均是ABP发生的高危因素（P<0.05）。当患者处于高TG水平时,高LPS组并发ABP的概率高于低LPS组（P<0.05）,在细小结石患者中,高LPS组并发ABP的概率高于低LPS组（P<0.05）。结论：依据结石形态学特征可对胆囊结石患者并发ABP的可能性作出早期的判断,同时血浆LPS水平升高是高TG以及细小胆囊结石患者易并发ABP的重要因素之一。     

早期非小细胞肺癌患者血清巨噬细胞抑制因子-1、趋化素与临床病理特征及预后的关系

目的:探讨早期非小细胞肺癌(NSCLC)患者血清巨噬细胞抑制因子-1(MIC-1)、趋化素(chemerin)水平与临床病理特征及预后的关系。方法:选择72例NSCLC患者(NSCLC组)、53例肺良性疾病患者(良性组)、50例体检健康人群(对照组),分别检测血清MIC-1、chemerin水平,分析血清MIC-1、chemerin水平与NSCLC患者临床病理参数的关系。Kaplan-Meier法分析不同血清MIC-1、chemerin水平NSCLC患者生存时间的差异,COX比例风险回归分析血清MIC-1、chemerin水平与NSCLC患者预后的关系。结果:NSCLC组患者血清MIC-1、chemerin水平高于良性组和对照组(P0.05)。血清MIC-1水平与NSCLC患者年龄、目前吸烟、肿瘤直径、TNM分期、分化程度、复发或转移、生存状态有关(P0.05),chemerin水平与NSCLC患者目前吸烟、TNM分期、复发或转移、生存状态有关(P0.05)。高MIC-1水平患者生存率低于低MIC-1水平患者(P0.05),高chemerin水平患者生存率低于低chemerin水平患者(P0.05)。COX比例风险回归分析结果显示:血清MIC-1、chemerin、TNM分期与NSCLC不良预后独立相关。结论:血清MIC-1、chemerin水平与NSCLC患者部分临床病理参数和预后相关,可作为早期NSCLC患者预后预测的潜在指标。     

A large nested association mapping population for breeding and quantitative trait locus mapping in Ethiopian durum wheat

The Ethiopian plateau hosts thousands of durum wheat (Triticum turgidum subsp. durum) farmer varieties (FV) with high adaptability and breeding potential. To harness their unique allelic diversity, we produced a large nested association mapping (NAM) population intercrossing fifty Ethiopian FVs with an international elite durum wheat variety (Asassa). The Ethiopian NAM population (EtNAM) is composed of fifty interconnected bi‐parental families, totalling 6280 recombinant inbred lines (RILs) that represent both a powerful quantitative trait loci (QTL) mapping tool, and a large pre‐breeding panel. Here, we discuss the molecular and phenotypic diversity of the EtNAM founder lines, then we use an array featuring 13 000 single nucleotide polymorphisms (SNPs) to characterize a subset of 1200 EtNAM RILs from 12 families. Finally, we test the usefulness of the population by mapping phenology traits and plant height using a genome wide association (GWA) approach. EtNAM RILs showed high allelic variation and a genetic makeup combining genetic diversity from Ethiopian FVs with the international durum wheat allele pool. EtNAM SNP data were projected on the fully sequenced AB genome of wild emmer wheat, and were used to estimate pairwise linkage disequilibrium (LD) measures that reported an LD decay distance of 7.4 Mb on average, and balanced founder contributions across EtNAM families. GWA analyses identified 11 genomic loci individually affecting up to 3 days in flowering time and more than 1.6 cm in height. We argue that the EtNAM is a powerful tool to support the production of new durum wheat varieties targeting local and global agriculture.