单纯疱疹病毒Ⅰ型新开放读码框UL57的鉴定

单纯疱疹病毒1型(Herpes simplex virus type 1,HSV-1)潜伏感染期间LATs的活跃转录可能与其启动子与增强子两侧的CTCF结合序列有关。本研究对位于UL56下游与LAT启动子上游之间并与CTCF结合序列重叠存在的一个新开放读码框(本研究中命名为UL57)进行了鉴定。首先利用HSV-1(F)细菌人工染色体(HSV-BAC)系统构建重组病毒HSV-EGFP-UL57,将EGFP序列插入UL57 5’端;然后分别通过Northern Blot和Western Blot检测EGFP标记的UL57的转录和表达;同时构建敲除UL57的重组病毒HSV-ΔUL57,观察UL57对病毒增殖的影响。结果显示,重组病毒HSV-EGFP-UL57感染HEp-2细胞17h后,EGFP探针检测到两条转录产物,其中1.8kb转录产物与预测大小相符;使用放线菌酮(Cycloheximide,CHX)阻断病毒即刻早期蛋白/早期蛋白合成后,UL57转录受到明显抑制。重组病毒HSV-EGFP-UL57感染Vero细胞后,9h可见融合蛋白表达,24h表达明显;融合蛋白分子量与预测大小(58kD)一致。病毒生长曲线显示,重组病毒HSV-EGFP-UL57及HSV-ΔUL57在Vero细胞中的增殖水平与HSV-1(F)基本一致。本研究表明,在HSV-1基因组(GenBank:GU734771.1)UL56下游与LAT启动子上游之间存在一个新开放读码框UL57(116 921bp～117 799bp),UL57可以进行转录,且其转录受病毒即刻早期蛋白/早期蛋白调控;转录产物可以翻译出融合蛋白,但表达水平较低。删除UL57对病毒增殖无明显影响。     

Thermal performance of quartz capillaries for vitrification

In this paper we report the thermal behavior of a new approach for vitrification. Thermal performance of traditional open pulled straws is compared with a new technique based on the combined use of quartz capillaries with slush nitrogen. This new method of vitrification achieved ultrafast cooling rates of 250,000 °C/min. As a result, a much lower concentration of cryoprotectant was needed to reach vitrification. In fact, a cryoprotectant solution typically used in oocyte slow freezing protocols was shown to remain transparent after cooling to liquid nitrogen temperatures indicating apparent “vitrification”. This approach offers a new and very promising technique for vitrification of cells using low levels of cryoprotectants.     

无充气腋窝入路腔镜下单侧甲状腺癌根治术与开放性手术的疗效比较及对免疫功能和颈部功能的影响

摘要 目的：比较无充气腋窝入路腔镜下单侧甲状腺癌根治术与开放性手术的疗效及对免疫功能和颈部功能的影响。方法：回顾性分析我院2020年8月～2021年8月期间收治的行单侧甲状腺癌根治术患者80例的临床资料。根据手术方式的不同将患者分为开放组（开放性手术）和腔镜组（无充气腋窝入路腔镜下单侧甲状腺癌根治术）,例数分别为37例和43例。对比两组疗效、免疫功能、颈部功能、美容学满意度和并发症情况。结果：与开放组相比,腔镜组术中出血量更少,手术时间、住院时间更长,引流液总量更多（P<0.05）,两组中央组淋巴结清扫数组间对比无统计学差异（P>0.05）。两组CD3⁺、CD4⁺、CD4⁺/CD8⁺下降,但腔镜组高于开放组;CD8+升高,但腔镜组低于开放组（P<0.05）。两组术后3d视觉疼痛模拟评分法（VAS）评分、颈部损伤指数对比,差异无统计学意义（P>0.05）。腔镜组吞咽障碍指数低于开放组（P<0.05）。腔镜组的总满意率高于开放组（P<0.05）。两组并发症发生率组间对比无统计学差异（P>0.05）。结论：与开放性手术治疗单侧甲状腺癌相比,无充气腋窝入路腔镜下单侧甲状腺癌根治术的手术时间和住院时间虽然延长,但其对患者免疫功能影响更轻,同时还可减轻患者吞咽障碍,获得更好的美容学满意度。     

Inbreeding and true seed in tetrasomic potato. IV. Synthetic cultivars

A true potato seed (TPS) synthetic cultivar from open pollination may broaden the genetic base in potato. A synthetic cultivar can also reduce the effect of inbreeding and maintain the productivity from generation to generation. Selected tetraploid parental genotypes, based on combining-ability tests, were chosen for isolated polycrosses to obtain tetraploid offspring. A series of experiments were undertaken to investigate the development of these TPS synthetic populations in two Peruvian locations: San Ramon, a rainfed, humid, mid-altitude environment, and La Molina, an arid, coastal environment under irrigation. Natural open-pollinated synthetics or control-mixture synthetics, involving two to six parents, had a similar tuber set but plant survival and tuber weight were higher in control synthetics including two or six parents. The results suggest that two-parent open-pollinated TPS synthetics may be a feasible option by selecting the right parents for the base population. A subsequent experiment showed that four-parent control synthetics (i.e. double crosses) were the best for tuber weight and size, followed by the two-parent control synthetics. Synthetic breeding populations could be shared with other breeders, who in cooperation with their local farmers may select promising genotypes for further cultivar release. Received: 15 December 2000 / Accepted: 4 April 2001     

Unrestricted modification search reveals lysine methylation as major modification induced by tissue formalin fixation and paraffin embedding

Formalin‐fixed paraffin‐embedded (FFPE) tissue is considered as an appropriate alternative to frozen/fresh tissue for proteomic analysis. Here we study formalin‐induced alternations on a proteome‐wide level. We compared LC‐MS/MS data of FFPE and frozen human kidney tissues by two methods. First, clustering analysis revealed that the biological variation is higher than the variation introduced by the two sample processing techniques and clusters formed in accordance with the biological tissue origin and not with the sample preservation method. Second, we combined open modification search and spectral counting to find modifications that are more abundant in FFPE samples compared to frozen samples. This analysis revealed lysine methylation (+14 Da) as the most frequent modification induced by FFPE preservation. We also detected a slight increase in methylene (+12 Da) and methylol (+30 Da) adducts as well as a putative modification of +58 Da, but they contribute less to the overall modification count. Subsequent SEQUEST analysis and X!Tandem searches of different datasets confirmed these trends. However, the modifications due to FFPE sample processing are a minor disturbance affecting 2–6% of all peptide‐spectrum matches and the peptides lists identified in FFPE and frozen tissues are still highly similar.     

生物化学检验技术实验教学与开放式综合实验设计初步效果探讨

目的:探讨生物化学检验技术实验教学(Experimental biochemistry inspection teaching,EBIT)与开放式综合实验设计(Open comprehensive experiment design,OCED)的初步效果,为EBIT教学提供新型选择方案。方法:选择2012届我们所带教医学检验专业的2个班级111例学生作为研究对象,分别归入观察组和对照组,其中观察组56例,对照组55例。对照组EBIT中予以传统的模式,观察组则采用OCED模式。对比两组实验成绩、教学效果的满意度。结果:观察组的合格率显著高于对照组,差异有统计学意义(P<0.05)。观察组对教学效果的满意度显著高于对照组,差异有统计学意义(P<0.05)。结论:OCED方式应用于EBIT过程中,可获得较好的教学效果,值得推荐。     

腹腔镜与开腹全系膜切除术治疗118 例直肠癌疗效研究

目的：探讨开腹与腹腔镜全系膜切除术(TME)治疗直肠癌的疗效及安全性。方法：选择本院收治的直肠癌患者118 例,随机 分为对照组和观察组,各为59 例。对照组行开腹直肠全系膜切除术,观察组行腔镜直肠全系膜切除根治术。比较两组术中效果、 淋巴结清扫效果、术后肠道功能恢复、手术风险和随访结果。结果：两组手术时间和淋巴结清扫数目无显著差异(P>0.05);观察组 切口长度和术中出血量显著少于对照组(P<0.05),肛门排气、肠胃功能恢复和总住院时间均显著短于对照组(P<0.05);观察组总并 发症率为5.08%,显著低于对照16.95%(P<0.05);观察组保肛率达到81.36%,显著高于对照组66.10%(P<0.05);两组2 年复发率和 转移率间差异无统计学意义(P>0.05)。结论：腹腔镜直肠癌TME 创伤小,根治率高,术后恢复快,并发症少,远期疗效与开腹手术 相近,综合疗效优于开腹根治术。     

tsORFdb: Theoretical Small Open Reading Frames (ORFs) database and massProphet: Peptide Mass Fingerprinting (PMF) tool for unknown small functional ORFs

Peptide mass fingerprinting (PMF) has become one of the most widely used methods for rapid identification of proteins in proteomics research. Many peaks, however, remain unassigned after PMF analysis, partly because of post-translational modification and the limited scope of protein sequences. Almost all PMF tools employ only known or predicted protein sequences and do not include open reading frames (ORFs) in the genome, which eliminates the chance of finding novel functional peptides. Unlike most tools that search protein sequences from known coding sequences, the tool we developed uses a database for theoretical small ORFs (tsORFs) and a PMF application using a tsORFs database (tsORFdb). The tsORFdb is a database for ORFeome that encompasses all potential tsORFs derived from whole genome sequences as well as the predicted ones. The massProphet system tries to extend the search scope to include the ORFeome using the tsORFdb. The tsORFdb and massProphet should be useful for proteomics research to give information about unknown small ORFs as well as predicted and registered proteins.