Microtiter micromass cultures of limb-bud mesenchymal cells

Summary A method is described for growing high-density micromass cultures of chick and mouse limb mesenchyme cells in 96-well microtiter plates (μTμM cultures). Rapid quantitative estimates of chondrogenic expression were obtained by automated spectrophotometric analysis of Alcian-blue-stained cartilage matrix extracts performed in the wells in which the cells had been grown. Quantitative estimates of myogenic expression were obtained similarly using anti-sarcomere myosin monoclonal antibody and modified ELISA techniques. This μTμM-ELISA method may be adapted for use with other antigens for which specific antibodies are available. These methods were used to compare cartilage and muscle differentiation in 1 to 4 d μTμM cultures grown in serum-containing (SCM) and defined (DM) media. The DM contains minimal additives (insulin, hydrocortisone, and in some cases, ascorbate or transferrin) and supports both chondrogenesis and myogenesis. The colorimetric analyses agree well with the morphologic appraisal of chondrogenesis and myogenesis. Similar numbers of cartilage nodules formed in all cultures, but in DM the nodules failed to enlarge; explaining the reduced matrix synthesis in DM as compared with SCM, and suggesting that nodule enlargement is a discrete, serum-dependent step. Studies of selected additives to DM show that transferrin enhances myogenesis, ascorbic acid enhances chondrogenesis, and retinoic acid inhibits chondrogenesis. Together, the μTμM system, in situ colorimetric assays of chondrogenesis and myogenesis, and DM will allow rapid prescreening of teratogens and screening of various bioactive compounds (e.g., hormones, growth factors, vitamins, adhesion factors) for effects on limb mesenchymal cell differentiation. This work was supported by grants RR08006-13 (DFP) and HD05505 and HD18577 (MS) from the National Institutes of Health, Bethesda, MD. MF-20 hybridoma supernatant was obtained from the Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa 52242 (maintained by NIH grant NO1-HD62915).     

检测特异性抗体分泌细胞(ASCs)的固相酶联免疫斑(ELISPOT)技术的建立

ＥＬＩＳＰＯＴ技术是目前国外评价疫苗免疫原性和免疫应答机理研究中最常使用的方法。本文用自制的ＮＣ膜２４孔板为固相支持物,建立了检测小鼠脾细胞中特异性ＡＳＣｓ的ＥＬＩＳＰＯＴ技术,在检测方法的特异性、结果的稳定性上均取得了满意的效果。并对该方法的某些实验条件进行了摸索。     

丙型肝炎病毒实验诊断技术的研究进展

目前,世界各国医学界均在积极探索非甲非乙型肝炎病毒的病原。随着分子克隆技术及特异性丙肝病毒抗原的出现,使特异性丙肝检测手段迅速发展,各种类型的诊断试剂盒不断推出,并在敏感性和待异性方面均有明显提高,日趋发展为简便、快速、可靠的方法。本文特就有关丙肝分子生物学及实验诊断技术的研究进展作一综述。     

Rat and human embryo and post-natal sera contain a potent endogenous competitor of estrogen-rat alpha-fetoprotein interactions

A highly active inhibitor of the binding of estrone and estradiol-17β to rat alpha-fetoprotein is demonstrated for the first time in embryo, immature and adult rat sera as well as in fetal and adult human sera. The competitive character and the narrow specificity of this inhibition effect is shown. The major compound responsible for this activity is isolated by successive column Sephadex LH₂₀ and thin layer chromatography : it is characterized as a nonpolar, nonphenolic, dialysable and thermostable substance, unreactive towards anti-estrone and anti-estradiol-17β anti-bodies. The possible biological role of an endogenous non-estrogen ligand of rodent fetoproteins is discussed.     

Stooping,crouching, and standing – Characterizing balance control strategies across postures

BackgroundWhile stooping and crouching postures are critical for many activities of daily living, little is known about the balance control mechanisms employed during these postures. Accordingly, the purpose of this study was to characterize the mechanisms driving net center of pressure (COP_Net) movement across three postures (standing, stooping, and crouching) and to investigate if control in each posture was influenced by time.MethodsTen young adults performed the three postures for 60 s each. Kinetic signals were collected via a force platform under each foot. To quantify mechanisms of control, correlations (Correl_LR) were calculated between the left and right COP trajectories in the anterior-posterior (AP) and medio-lateral (ML) directions. To examine the potential effects of time on balance control strategies, outcomes during the first 30 s were compared to the last 30 s.ResultsCorrel_LR values did not differ across postures (AP: p = 0.395; ML: p = 0.647). Further, there were no main effects of time on Correl_LR (AP: p = 0.976; ML: p = 0.105). A significant posture-time interaction was observed in the ML direction (p = 0.045) characterized by 35% decreases in Correl_LR over time for stooping (p = 0.022).ConclusionThe dominant controllers of sway (i.e., AP: ankle plantar/dorsi flexors; ML: hip load/unload mechanism) are similar across quiet stance stooping, and crouching. Changes in ML control strategies over time suggests that fatigue could affect prolonged stooping more so than crouching or standing.     

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

We describe the complete process of AcroPrep Advance Filter Plates for 96 plasmid preparations, starting from prokaryotic culture and ending with high purity DNA. Based on multi-well filtration for bacterial lysate clearance and DNA purification, this method creates a streamlined process for plasmid preparation. Filter plates containing silica-based media can easily be processed by vacuum filtration or centrifuge to yield appreciable quantities of plasmid DNA. Quantitative analyses determine the purified plasmid DNA is consistently of high quality with average OD_260/280 ratios of 1.97. Overall, plasmid yields offer more pure DNA for downstream applications, such as sequencing and cloning. This streamlined method of using AcroPrep Advance Filter Plates allows for manual, semi-automated or fully-automated processing.     

后侧手术入路、锁定钢板固定治疗胫骨平台后髁骨折的临床疗效

目的：探讨采用膝关节后内和后外侧切口、锁定钢板固定治疗胫骨平台后髁骨折的临床疗效。方法：选取2009年1月～2011年12月来我院治疗的13例胫骨平台后髁骨折患者作为研究对象,所有患者采用后内和后外侧切口、锁定钢板固定治疗,观察术后骨折愈合情况,采用Rasmussen放射学评分评定骨折复位情况,采用HSS评分系统评定膝关节功能疗效。结果：术后随访6～24个月,平均13．6个月,所有患者骨折均于16周内愈合。Rasmussen放射学评分15～18分,平均17．4分,其中优8例,良3例,可2例,优良率为84．7％。末次随访时膝关节功能HSS评分73~97分,平均91．4分,其中获优7例,良5例,可1例,优良率为92．3％。结论：采用膝关节后内和（或）后外侧切口、锁定钢板固定治疗胫骨平台后髁骨折具有操作简单、显露清晰、直视下复位骨折及关节面、固定牢靠、利于早期功能锻炼、临床疗效确切、并发症少等优点。     

肱骨近端锁定加压钢板与解剖钢板治疗老年肱骨外科颈骨折的临床疗效

目的:研究肱骨近端锁定加压钢板与解剖钢板治疗老年肱骨外科颈骨折的临床疗效。方法:选取92例老年肱骨外科颈骨折患者作为研究对象。根据数字法随机将患者分成观察组及对照组,各含46例。其中对照组予以解剖钢板术式治疗,观察组则予以肱骨近端的锁定加压钢板术式治疗。对比两组不同治法的疗效,两组患者的手术相关指标,两组患者治疗后的肩关节活动度以及两组患者治疗前后的Constant评分及VAS评分。结果:观察组的优良率是89.13%(41/46),显著高于对照组的76.09%(35/46),差异有统计学意义(P0.05)。观察组的术程、手术出血量以及住院时间和骨折的愈合时间均分别显著少于对照组,差异均有统计学意义(均P0.05)。观察组的肩关节活动度显著大于对照组,差均有统计学意义(P0.05)。治疗后观察组的Constant评分及VAS评分均分别显著高于对照组,差异均有统计学意义(均P0.05)。结论:利用肱骨近端的锁定加压钢板术式治疗FSNH,能够帮助患者获得更为积极的临床预后,值得在临床治疗过程中推广应用。     

锁定加压钢板和动力加压钢板治疗肱骨中下段骨折的临床比较研究

目的:对比锁定加压钢板与动力加压钢板治疗肱骨中下段骨折患者的临床疗效。方法:抽取我院2010年8月~2013年12月收治的肱骨中下段骨折患者76例,按照随机数字表法分为锁定组(锁定加压钢板治疗)与动力组(动力加压钢板治疗),每组38例,随访1年。对比两组患者临床指标,治疗效果及并发症发生率。结果:锁定组患者手术时间、骨折愈合时间及住院时间均小于动力组,差异均有统计学意义(P0.05);锁定组患者优良率为89.47%,显著高于动力组的71.05%,差异有统计学意义(P0.05);锁定组的并发症发生率为10.53%,显著低于动力组的31.58%,差异有统计学意义(P0.05)。结论:锁定加压钢板应用于治疗肱骨中下段骨折患者疗效优于动力加压钢板治疗,它具有创伤小、恢复快的优点,且并发症发生率较低,值得推广。     

微创经皮钢板联合锁定加压钢板固定术治疗胫骨干骺端骨折的临床疗效

目的:探讨微创经皮钢板固定技术(MIPPO)与锁定加压钢板(LCP)治疗胫骨干骺端骨折的临床疗效。方法:选择2013年6月-2015年6月在我院接受手术治疗的胫骨干骺端骨折患者103例作为研究对象,根据手术方法不同将所选患者分为三组。其中,联合手术组(35例)患者采用MIPPO联合LCP内固定术治疗,外固定组(33例)患者采用超关节外固定支架术治疗,内固定术组(35例)患者采用切开复位钢板内固定术治疗。观察并比较三种手术方法的临床效果及对患者骨关节功能的影响。结果:与外固定术组及内固定术组比较,联合手术组患者的手术时间短,术中出血量少,骨折愈合时间早,差异具有统计学意义(P0.05);联合手术组患者的手术优良率高于外固定组和内固定组,差异有统计学意义(P0.05);内固定组手术优良率高于外固定组,差异具有统计学意义(P0.05);联合手术组患者术后并发症的发生率低于外固定组及内固定组,差异具有统计学意义(P0.05);外固定组与内固定组术后并发症的发生率比较,差异无统计学意义(P0.05)。结论:MIPPO联合LCP内固定治疗胫骨干骺端骨折具有手术切口小、术中出血量少、术后并发症发生率低、骨折愈合快、关节功能恢复较好等优点,是治疗胫骨干骺端骨折的理想方法,值得临床推广应用。