A Cocktail ELISA Semi‐nested RT‐PCR Assay to Detect Bean pod mottle virus in Soya bean Seeds

Bean pod mottle virus (BPMV) has been identified as an important pathogen for plant quarantine in China because large quantities of soya bean seeds (approximately 7 × 10⁷ tons) are imported annually. To develop a practical detection programme for BPMV, a cocktail enzyme‐linked immunosorbent assay (ELISA) nested RT‐PCR using a combination of serological and molecular methods was designed for soya bean seeds. The single‐vessel detection assay was performed in a 96‐well ELISA plate, which served as a carrier for the subsequent nested RT‐PCR assay. Assay specificity was demonstrated by the production of the expected 330‐ and 296‐bp bands using the external and internal primers, respectively. This method was 10⁴‐fold more sensitive than immunocapture‐RT‐PCR (IC‐RT‐PCR). In particular, it is important to note that this assay resulted in successful micro‐extraction from soya bean seeds and combined the advantages of each individual technique. The cocktail ELISA nested RT‐PCR is a specific, sensitive, rapid and economical procedure to rapidly identify and characterize BPMV and could be suitable for both primary‐level platforms and laboratories.     

外阴阴道假丝酵母菌病伴发支原体及沙眼衣原体感染的研究

为研究外阴阴道假丝酵母菌病(Vulvovaginal Candidiasis,VVC)患者合并支原体和衣原体感染情况,对86例复发性VVC(recurrent Vulvovaginal Candidiasis,RVVC)患者、219例单纯性VVC患者以及健康妇女200例,分别进行解脲脲原体(Ureaplasma urealyticum,Uu)、人型支原体(mycoplasma hominis,Mh)和沙眼衣原体(Chlamydia trachomatis,CT)检测;所有VVC病例均进行真菌培养。RVVC组和单纯VVC组与对照组之间Uu、Mh及Uu混合Mh阳性率比较,差异均有统计学意义(P0.05)。而三组之间CT和CT混合支原体阳性率比较,均无显著性差异(P0.05)。真菌培养阳性组与阴性组之间,Uu阳性率比较有显著性差异(P0.05),而CT阳性率比较无显著性差异(P0.05)。结果表明,VVC患者合并Uu感染较正常人群明显增高,假丝酵母菌与Uu混合感染可能导致VVC的发生和复发。     

高校病理生理学双语教学探析

双语教学作为蕴含着全新教育理念的教学形式,是我国高等教育改革的一项新措施,适应新时代背景下社会对高质量人才的需求。探析双语教学引入高校病理生理学中的可行意义、病理生理学双语教学中存在的问题、病理生理学双语教学的改进路径,有利于高校病理生理学双语教学的实践,对高校教育改革及医学专业人才培养起到积极的推动作用。     

高校互动式教学模式探讨

互动式教学模式是指以学生为主体,立足于培养学生创新能力的一种教学模式。高等医学院校是高素质医学人才培养的主要单位,研究高校互动式教学模式的内涵及医学意义、病理生理学中互动式教学模式的应用、互动式教学模式的制约瓶颈、探究高校互动式教学模式的实施策略有利于促进高校互动式教学模式的创新及医学人才培养质量的提高。     

藜科植物藜和灰绿藜实时荧光定量PCR内参基因的选择

选择合适的内参基因是实时荧光定量PCR(qRT-PCR)研究的关键,目前对藜科耐盐(盐生)植物胁迫相关基因的表达分析中所用内参基因的报道较为有限.该研究利用GeNorm、NormFinder和BestKeeper 3个内参基因分析软件,对已选择过的β-TUBULIN、β-ACTIN、GAPDH 3个常用候选内参基因进行了比较分析,筛选出在NaCl和PEG胁迫下藜和灰绿藜中表达相对稳定的内参基因.结果表明:GeNorm、NormFinder内参软件分析在NaCl和PEG胁迫下,GAPDH是藜和灰绿藜中均共同稳定表达的内参基因,同时在藜和灰绿藜中也有各自表达较稳定的内参基因,β-ACTIN在藜中稳定表达,β-TUBULIN则在灰绿藜中稳定表达.对相同科不同种的植物内参基因表达差异进行比较,内参基因在相同科中具有相同稳定表达的内参;对相同胁迫下两种不同植物内参基因表达稳定性进行分析,内参基因的选择需根据实际的实验材料和实验条件而定.基于3个分析软件对以上3个常用内参基因的分析结果,初步确定了在藜科植物藜和灰绿藜中相对稳定的内参基因,为藜和灰绿藜胁迫相关基因的定量表达分析提供了参考依据.     

太空诱变玉米核不育突变体矮化的遗传及外施赤霉素分析

为了解太空诱变玉米核不育突变体矮化的遗传规律和原因,该研究以不育突变体为母本,自交系178、478为父本,对测交 F1、F2群体进行育性鉴定和株高分析,对 F2可育株进行基因型和株高分析,对姊妹交后代分离群体进行育性鉴定和株高、雄穗长度、节间数、节间长度分析,同时,还对姊妹交后代分离群体进行施赤霉素处理,调查育性和株高的变化。结果表明：178和478背景下的 F1表现出与测交母本一样的极显著差异;在178和478核背景下的 F2中,不育株株高极显著矮于可育株,两核背景下的不育株间株高差异不显著,而可育株间株高差异极显著;F2中纯合和杂合可育株的株高差异不显著;姊妹交后代分离群体中不育株株高、雄穗长度、节间数和节间长度极显著小于可育株;外施赤霉素的不育株在苗期表现出对赤霉素一定的敏感性,但株高最终未恢复正常高度。因此,得出该突变体矮化表现稳定,与不育性状并存,且不受细胞核背景的影响;核不育基因对植株株高的矮化无剂量效应;突变体的矮化与雄穗长度、节间数和节间长度有关;突变体不完全属于赤霉素不敏感型,其矮化并不是单一缺乏赤霉素而引起。该研究结果为认识太空诱变玉米核不育突变体矮化的遗传和生理机制提供了参考。     

油茶叶片营养元素、叶脉密度及生理指标随树龄变化规律及其与产量的相关性

该研究关注了不同树龄油茶叶片的营养元素含量、叶脉密度及生理指标,旨在了解它们随树龄的变化规律及其对产量的影响。结果表明：(1)不同树龄间金属或稀土元素含量差异显著;大部分元素随树龄的增加表现为先增后减的变化趋势,但不同元素达到最大含量的树龄不尽相同;(2)叶脉呈网络状分布,根据叶脉直径大小分为粗、中、细三个等级;粗、中叶脉密度随树龄的变化不明显,细叶脉密度随树龄呈先增后减变化趋势。(3)叶绿素、MDA、可溶性糖、可溶性蛋白质含量以及POD活性等生理指标均随树龄的变化,表现出各自变化规律。(4)叶绿素含量、POD活性、稀土元素总量以及细叶脉密度与产量表现出明显正相关关系(P<0.05, P<0.01)。该研究结果丰富了油茶的基础资料,可为油茶的栽培与管理提供些许参考,并提示人们可以利用叶片的部分营养、生理、结构指标来跟踪监测植株状态和预测产量。     

Role of Phosphotyrosine Interaction Domain Containing 1 in Porcine Intramuscular Preadipocyte Proliferation and Differentiation

Phosphotyrosine interaction domain containing 1 (PID1), a recently identified gene involved in obesity-associated insulin resistance, plays an important role in fat deposition. However, its effect on porcine intramuscular preadipocyte proliferation and differentiation remains poorly understood. In this study, the plasmid pcDNA3.1(+)-pPID1 was transfected into porcine intramuscular preadipocytes with Lipofectamine 3000 reagent to over-express porcine PID1 (pPID1). Over-expression of pPID1 significantly promoted porcine intramuscular preadipocyte proliferation. Expression of pPID1 mRNA was significantly increased upon porcine intramuscular preadipocyte differentiation. Indirect fluorescent immunocytochemistry demonstrated that pPID1 protein was localized predominantly in the nucleus of porcine intramuscular preadipocyte. The mRNA levels of peroxisome proliferators-activated receptor γ, CCAAT/enhancer binding protein α and lipoprotein lipase were significantly increased by pPID1 over-expression. Over-expression of pPID1 also led to an increase in lipid accumulation which was detected by Oil Red O staining, and significantly increased the intramuscular triacylglycerol content. These results indicate that pPID1 may play a role in enhancing porcine intramuscular preadipocyte proliferation and differentiation.