室旁核注射胰高血糖素样肽-1对不同病程糖尿病大鼠胃排空的影响（英文）

目的：研究下丘脑室旁核（paraventricular nucleus,PVN）注射胰高血糖素样肽-1（GLP-1）对糖尿病早期大鼠胃排空的影响,并探讨其相关作用机制。方法：60只清洁级雄性Wistar大鼠随机分为正常对照组（NC组）,糖尿病组（DM组）,GLP-1干预组（GLP-1组）,每组各20只,后两组腹腔注射链脲佐菌素（STZ）制备糖尿病模型,分别于注射STZ2周、6周后每组随机取半数进行实验,实验前于无菌条件下大鼠一侧下丘脑PVN区埋置套管,GLP-1组经套管注入GLP-1,NC组及DM组注入等体积生理盐水。酚红灌胃法检测胃排空率,酶联免疫吸附法（ELISA）测定血浆GLP-1浓度,半定量RT-PCR法测定胃窦、胃底GLP-1RmRNA表达。结果：注射STZ 2周后,DM组较NC组胃排空率显著升高（P〈0.01）。GLP-1组胃排空率低于DM组（P〈0.01）,血浆GLP-1浓度高于DM组及NC组（P均〈0.05）,胃窦GLP-1RmRNA表达明显高于DM组、NC组（P均〈0.01）。注射STZ 6周后,DM组胃排空率高于NC组（P〈0.01）。GLP-1组较DM组胃排空率显著降低（P〈0.01）,血浆GLP-1浓度、胃窦GLP-1RmRNA表达显著高于DM组、NC组（P均〈0.01）。结论：下丘脑PVN区注射GLP-1后,可减慢糖尿病大鼠初期加速的胃排空,原因可能与血浆GLP-1浓度及胃窦GLP-1RmRNA表达增加有关。     

生物化学“2463”课堂教学设计与实践

生物化学是生命科学领域学科专业重要基础课之一,也是"教"与"学"的两难课程之一。基于国内外教学理论和农学专业课程多年的实践教学经验,构建了生物化学课程的"2463"教学模式:在师生双主体的前提下,在教学内容、教学方法、教学手段和教学评价四个层面开展了六个目标方向的实践探索,即教学内容的前沿化、生活化和人文化,教学方式的多元化,教学手段的信息化和教学评价的过程化,而且每个目标方向至少进行三个路径的实践探索。该教学模式经过五轮的教学实践,取得了良好的教学效果。     

潮霉素A的生物合成研究进展

潮霉素A是一种从吸水链霉菌中发现的具有广谱生物学活性的抗生素。它在吸水链霉菌Streptomy-ces hygroscopicus NRRL 2388中的生物合成基因簇已被克隆并测序,其生物合成机制、遗传操作等方面的研究也取得了一定的进展。就潮霉素A的化学结构、生物合成基因簇的组织结构、生物合成和抗性机制等方面的研究进展进行综述。     

The cost of virulence: retarded growth of Salmonella Typhimurium cells expressing type III secretion system 1

Virulence factors generally enhance a pathogen's fitness and thereby foster transmission. However, most studies of pathogen fitness have been performed by averaging the phenotypes over large populations. Here, we have analyzed the fitness costs of virulence factor expression by Salmonella enterica subspecies I serovar Typhimurium in simple culture experiments. The type III secretion system ttss-1, a cardinal virulence factor for eliciting Salmonella diarrhea, is expressed by just a fraction of the S. Typhimurium population, yielding a mixture of cells that either express ttss-1 (TTSS-1(+) phenotype) or not (TTSS-1(-) phenotype). Here, we studied in vitro the TTSS-1(+) phenotype at the single cell level using fluorescent protein reporters. The regulator hilA controlled the fraction of TTSS-1+ individuals and their ttss-1 expression level. Strikingly, cells of the TTSS-1(+) phenotype grew slower than cells of the TTSS-1(-) phenotype. The growth retardation was at least partially attributable to the expression of TTSS-1 effector and/or translocon proteins. In spite of this growth penalty, the TTSS-1(+) subpopulation increased from <10% to approx. 60% during the late logarithmic growth phase of an LB batch culture. This was attributable to an increasing initiation rate of ttss-1 expression, in response to environmental cues accumulating during this growth phase, as shown by experimental data and mathematical modeling. Finally, hilA and hilD mutants, which form only fast-growing TTSS-1(-) cells, outcompeted wild type S. Typhimurium in mixed cultures. Our data demonstrated that virulence factor expression imposes a growth penalty in a non-host environment. This raises important questions about compensating mechanisms during host infection which ensure successful propagation of the genotype.     

血浆uPA、uPAR在乳腺癌分子亚型中的表达及临床意义

目的:探讨血浆uPA、uPAR在不同分子亚型乳腺癌中的表达水平及其对乳腺癌患者治疗、预后等的临床意义。方法:采用免疫组化ELISA方法测定的女性乳腺癌初治患者86例的血浆uPA、uPAR水平,所有患者均经组织病理学确诊,以患者的临床病理学资料提供的免疫组化结果为基础进行分子分型,结合二者进行分析。结果:血浆uPA在乳腺癌不同分子亚型中表达有统计学差异(P<0.01),uPAR在乳腺癌不同分子亚型中表达有统计学差异(P<0.05);乳腺癌患者血浆uPA和uPAR呈显著正相关(P<0.01,Pearson相关系数r=0.735)。结论:乳腺癌患者血浆uPA和uPAR的水平与分子亚型密切相关,他们和分子亚型联合,可能对乳腺癌个体化治疗及判定预后有指导意义。     

布拉酵母联合舍曲林治疗产后抑郁症的疗效观察

目的探讨布拉酵母菌联合舍曲林对产后抑郁症(PPD)的临床疗效及安全性。方法选择2016年1月至2018年12月我院104例PPD患者为研究对象。入选患者随机分为观察组和对照组各52例,两组患者均给予舍曲林50 mg/次,1次/d,口服。观察组患者同时联用布拉酵母0.5 g/次,2次/d,口服,8周为一疗程。治疗前后检测患者孕酮及雌二醇水平,并于治疗前及治疗第4周、第8周时采用汉密顿抑郁量表(HAMD)评价患者治疗结果、临床疗效及安全性。结果治疗后,两组患者HAMD评分与治疗前比较均降低(t=8.162,P0.001;t=17.916,P0.001;t=16.995,P0.001;t=28.683,P0.001),且观察组患者降低幅度大于对照组(t=7.741,P0.001;t=13.073,P0.001)。观察组患者临床总有效率(90.38%)明显高于对照组(69.23%)。治疗后观察组患者孕酮水平低于对照组,雌二醇水平高于对照组,差异均有统计学意义(t=10.774,P0.001;t=7.239,P0.001)。两组母婴无明显不良反应。结论布拉酵母联合舍曲林对产后抑郁症的临床疗效明显,安全性高,值得临床推广。     

Rab GDP解离抑制因子的定位和表达

目的:构建含人GDI1和GDI2基因的真核表达载体进行定位和蛋白表达研究。方法:用PCR从U251细胞cDNA克隆GDI1和GDI2基因,构建真核表达载体pEGFP-N2-GDI1和pEGFP-N2-GDI2,转染HEK293T细胞。荧光显微镜观察GDI1和GDI2蛋白的细胞内定位,再通过SDS-PAGE和Western blotting鉴定蛋白的表达。结果:成功克隆到1 341bp和1 335bp的人源GDI1和GDI2基因,并准确插入真核表达载体pEGFP-N2中,荧光观察这两个蛋白定位到细胞浆中,并能利用标签抗体检测到GDI1和GDI2的表达。结论:GDI1和GDI2能够定位到细胞浆,并能通过Western blotting检测,为进一步研究GDI1和GDI2的功能奠定了基础。     

2型糖尿病患者心率变异性与心率减速力的相关性及其对自主神经功能的评估价值

目的:探讨2型糖尿病(T2DM)患者心率变异性(HRV)与心率减速力(DC)的相关性及其对自主神经功能的评估价值。方法:选取2018年1月～2019年12月期间合肥市第一人民医院收治的120例T2DM患者为研究对象,纳入实验组,其中单纯T2DM患者69例(单纯T2DM组),T2DM伴周围神经病变(DPN)患者51例(DPN组),另选同期在我院进行心功能检查的健康志愿者120例作为对照组。所有受试者均进行24h动态心电图检查,根据检查结果计算HRV值[总标准差(SDNN)、两个相邻RR间期互差(PNN50)、差值均方根(RMSSD)]和DC值,根据各组受试者的DC值,统计对比各组的猝死风险,并分析T2DM患者DC值与HRV各项指标的相关性。结果:对照组、单纯T2DM组、DPN组的SDNN、PNN50、RMSSD、DC值依次降低(均P0.05)。对照组、单纯T2DM组、DPN组各猝死风险等级分布整体比较差异有统计学意义(P0.05),单纯T2DM组、DPN组猝死高危比例显著高于对照组,且DPN组猝死高危比例高于单纯T2DM组(均P0.05)。经Pearson相关分析,T2DM患者的DC值与SDNN、PNN50、RMSSD呈明显的正相关(P0.05)。结论:T2DM患者的DC值、HRV指标均低于正常人群,合并DPN的T2DM患者猝死风险明显提高,DC值与HRV指标间存在明显的正相关,可作为T2DM患者自主神经功能状态评估的重要指标。     

Live covisualization of competing adeno-associated virus and herpes simplex virus type 1 DNA replication: molecular mechanisms of interaction

We performed live cell visualization assays to directly assess the interaction between competing adeno-associated virus (AAV) and herpes simplex virus type 1 (HSV-1) DNA replication. Our studies reveal the formation of separate AAV and HSV-1 replication compartments and the inhibition of HSV-1 replication compartment formation in the presence of AAV. AAV Rep is recruited into AAV replication compartments but not into those of HSV-1, while the single-stranded DNA-binding protein HSV-1 ICP8 is recruited into both AAV and HSV-1 replication compartments, although with differential staining patterns. Slot blot analysis of coinfected cells revealed a dose-dependent inhibition of HSV-1 DNA replication by wild-type AAV but not by rep-negative recombinant AAV. Consistent with this, Western blot analysis indicated that wild-type AAV affects the levels of the HSV-1 immediate-early protein ICP4 and the early protein ICP8 only modestly but strongly inhibits the accumulation of the late proteins VP16 and gC. Furthermore, we demonstrate that the presence of Rep in the absence of AAV DNA replication is sufficient for the inhibition of HSV-1. In particular, Rep68/78 proteins severely inhibit the formation of mature HSV-1 replication compartments and lead to the accumulation of ICP8 at sites of cellular DNA synthesis, a phenomenon previously observed in the presence of viral polymerase inhibitors. Taken together, our results suggest that AAV and HSV-1 replicate in separate compartments and that AAV Rep inhibits HSV-1 at the level of DNA replication.     

Collagen synthesis of articular cartilage explants in response to frequency of cyclic mechanical loading

Articular cartilage in vivo experiences the effects of both cell-regulatory proteins and mechanical forces. This study has addressed the hypothesis that the frequency of intermittently or continuously applied mechanical loads is a critical parameter in the regulation of chondrocyte collagen biosynthesis. Cyclic compressive pressure was applied intermittently to bovine articular cartilage explants by using a sinusoidal waveform of 0.1–1.0 Hz frequency with a peak stress of 0.5 MPa for a period of 5–20 s followed by a load-free period of 10–1,000 s. These loading protocols were repeated for a total duration of 6 days. In separate experiments, cyclic loading was continuously applied by using a sinusoidal waveform of 0.001–0.5 Hz frequency and a peak stress of 1.0 MPa for a period of 3 days. Unloaded cartilage discs of the same condyle were cultured in identically constructed loading chambers and served as controls. We report quantitative data showing that (1) no correlation exists between the relative rate of collagen synthesis expressed as the proportion of newly synthesized collagen among newly made proteins and either the frequency of intermittently or continuously applied loads or the overall time cartilage is actively loaded, and (2) individual protocols of intermittently applied loads can reduce the relative rate of collagen synthesis and increase the water content, whereas (3) continuously applied cyclic loads always suppress the relative rate of collagen synthesis compared with that of unloaded control specimens. The results provide further experimental evidence that collagen metabolism is difficult to manipulate by mechanical stimuli. This is physiologically important for the maintainance of the material properties of collagen in view of the heavy mechanical demands made upon it. Moreover, the unaltered or reduced collagen synthesis of cartilage explants might reflect more closely the metabolism of normal or early human osteoarthritic cartilage.This work was supported by the Federal Ministry of Education and Research (BMBF no. 0311058) and by the foundation S.E.T.