肥大型骨不连断端组织成骨潜能的实验研究

目的　观察肥大型骨不连断端及其周围区域组织的结构特点,评估其成骨潜能。 方法　收集2009年至2012年因肥大型骨不连在我院行手术治疗患者的骨不连断端标本共25例,通过光学显微镜及透射电镜观察不同区域的组织结构特点和微观特征。采用免疫组化检测BMP-2的表达,在高倍镜(400倍)下对阳性细胞进行观察并计数,并作统计分析。 结果　肥大型骨不连断端区域呈现出不同的组织类型,中心区主要是纤维化组织,其内可见大量的成纤维样细胞;边缘区可见大量的软骨细胞,可见少量的新生血管形成;骨痂区见有一定量的成骨细胞及丰富的新生血管。免疫组化染色显示肥大型骨不连断端中心区、周围区及骨痂区组织BMP-2的表达明显不同（P<0.05）。 结论　肥大型骨不连断端区域的组织及细胞类型呈现出不同的特点,BMP-2含量减少使得不同区域的成骨潜能不同。     

丹参果糖磷酸酶基因的克隆和功能研究

丹参是著名的药用植物,在中国、日本、美国和欧洲国家被广泛地用于心脑血管系统疾病的治疗.笔者首次从丹参中克隆出了一个新的果糖磷酸酶基因,并将其命名为SmFBA,GenBank编号为FJ540907.SmFBA的cDNA全长含有1 390个核苷酸,包含一个完整的1 065 bp的开放阅读框,编码355个氨基酸残基.SmFBA基因全长含有3个外显子和2个内含子.生物信息学分析显示SmFBA蛋白预测的等电点为5.60,预测的分子量为37.78 ku,和其他植物物种中果糖磷酸酶具有很高的序列同源性.用Southern杂交技术显示SmFBA在丹参基因组中呈低拷贝.该基因在丹参根、茎、叶等器官都表达,根中表达量最高.体外重组表达的SmFBA在大肠杆菌中具有酶活性,并且能够提高大肠杆菌的耐盐性.这项研究进一步拓展了人们对高等植物糖酵解途径的认识.     

刺五加环阿屯醇合酶基因的克隆及其表达分析

目的克隆刺五加的环阿屯醇合酶(cycloartenol synthase,CAS)基因,并对其进行生物信息学和表达分析。方法采用cDNA末端快速扩增(rapid amplification of cDNAends,RACE)技术克隆刺五加CAS基因的全长cDNA序列。运用生物信息学方法对该基因进行分析,预测其编码蛋白的结构与功能,并通过RT-PCR法检测CAS在不同生长发育时期和不同器官中的表达情况。结果刺五加CAS基因的cDNA全长为2 758 bp,开放阅读框长2 277 bp,编码758个氨基酸的蛋白,包含三萜合成酶的标志性序列。CAS蛋白无跨膜区域,定位于细胞质中。RT-PCR的结果显示,刺五加CAS基因在各时期和器官中均有表达,但表达量具有显著差异(P<0.05)。其中果实基本成熟期的表达量最高,是最低量萌芽期的1.56倍,各器官中,叶片的表达量最高,是最低量叶柄的1.37倍。结论首次分离并报道了刺五加的CAS cDNA克隆,并证实其在不同生长发育时期和不同器官中的表达量不同,为进一步研究CAS对刺五加皂苷量的影响和表达调控奠定基础。     

Length-tension relation of cat heart muscle studied by a segment-control method

Summary We studied the effects of the damaged ends on the length-tension relationship of the cat right ventricular papillary muscle during twitch contraction (30/min) using a newly developed segment-control system. The segment length (SL), defined as the distance between two thin black films attached to the central surface of the muscle with silicone grease, was measured by a Hamamatsu Photonics Width Analyzer (C1170) and Camera (C1000) and controlled with a servoactuator. The marker distance was determined to be either the central 24%–30% (short marker distance) or 47%–56% (long marker distance) of the muscle length (ML) at which the maximum developed twitch tension was observed (MLmax). The muscle was lengthened and shortened in a triangular fashion under both ML and SL isometric twitch modes over a period of 250s. The relationship between ML or SL and developed tension (DT, total minus resting tension) was obtained from the twitch tension data during the lengthening. The DT in the SL isometric twitch was always greater than that in the ML isometric twitch over the range of lengths studied. The slope of the SL-DT relation curve was much less steep than that of the ascending limb of the ML-DT relation curve. The gradient of the SL-DT relationship curve was greater for the long marker than for the short marker distance. The percentage shortening of SL in the ML isometric twitch increased with decreasing ML; it was greater for the short marker than for the long marker distance, particularly at shorter MLs. These results suggest that even the central part of the excised cardiac muscle has inhomogeneous contractile properties.     

家族性急性髓系白血病相关新基因FAMLF cDNA全长的克隆及其生物功能分析

目的 克隆家族性急性髓系白血病(AML)相关新基因cDNA全长,在分子水平上探讨急性白血病发生发展的机制.方法 以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87(GenBank注册号CV973101)为基础,应用cDNA末端快速扩增法(RACE)克隆其全长cDNA,应用生物信息学对其功能进行初步分析,应用一步法半定量RT-PCR检测其在AML患者及正常人中的表达.结果 获得了家族性AML相关新基因FAMLF的cDNA全长,该基因定位于染色体lq31.3,cDNA全长2313 bp,开放读码框(ORF)为249 bp,编码82个氨基酸的蛋白质,含有信号肽,富含亮氨酸重复单位(LRR_SD22),内在固有无序结构域等功能区.Blast检索为功能未知的新基因,已被GenBank收录,并命名为FAMLF,核酸注册号为EF413001,蛋白质注册号为ABN58747.新基因FAMLF在AML患者中的表达明显高于正常人,差异有统计学意义(2.61±0.66 vs 0.97±0.51,P<0.01).结论 成功获得一个家族性AML相关新基因FAMLF cDNA全长,FAMLF基因在AML中高表达,可能是具有一定生物功能的新基因.     

肢端皮肤角化过度增生症的手术治疗

目的介绍肢端皮肤角化过度增生症10例的治疗体会,其中掌跖角化病9例,疣状表皮结构不良1例。方法采用病变皮肤切除皮肤移植术治疗,其中两例有深部组织外露和放射性溃疡选用皮瓣修复,其余行皮片移植。结果 10例均获满意效果,追踪观察8年以上,无复发,能保持接近正常的功能活动。结论手部以皮片效果最佳,足底应用皮瓣修复,耐磨性较皮片为好,但因皮瓣缺乏纤维隔结构,走路时皮瓣与深部组织间产生相对滑动,也容易造成破溃,必须注意妥为保护。     

猪-人移植细胞性排斥反应相关候选基因pOSR1的克隆和鉴定

为寻找与异种移植细胞性排斥反应相关的新基因 ,在抑制消减杂交 (SSH )基础上 ,对一个与人氧化应激反应 1(OSR1)基因同源的猪内皮细胞上调表达基因片段CS3作进一步研究。半定量RT PCR结果证实 ,在人PBMC作用后 ,该基因表达的确上调。应用快速cDNA末端扩增 (RACE )方法 ,获得了猪OSR1(pOSR1)的全长cDNA序列。pOSR1cDNA序列全长 4 333个碱基 ,开读框架长 15 90个碱基 ,编码 5 2 9个氨基酸。与人OSR1(hOSR1)相比 ,编码区核酸一致性为 92 8% ,氨基酸一致性为 95 5 %。GenBank登录号为AY2 7135 6。其功能研究正在进行中。     

INFLUENCE OF LOWERED TEMPERATURE UPON THE RESPONSE OF PROSTATIC AND EPIDIDYMAL PORTION OF THE RAT ISOLATED VAS DEFERENS TO FIELD STIMULATION

This paper reports the effects of lowering (bath) temperature upon contractile responses of prostatic and epididymal ends of the rat isolated vas deferens to electrical field stimulation. Responses to field stimulation with single pulses at 60 s intervals were potentiated by lowering the bath temperature. This effect was maximal at temperatures of 25-27 degrees C and was reduced in preparations treated with phentolamine (5 mumol/l). Field stimulation with trains of 50 pulses at 5 Hz every 5 min produced biphasic contractions of preparations. The initial, twitch-like component, prominent at the prostatic end and resistant to blockade by phentolamine (5 mumol/l) was little affected by lowering the bath temperature from 37.5 degrees C to 19 degrees C. In contrast, the second, more prolonged, component of the response to pulse trains, which was prominent at the epididymal end of the tissue was decreased by lowering the bath temperature and by phentolamine. In the epididymal segment, responses to exogenous noradrenaline were enhanced by lowering the bath temperature. It is concluded that lowering the bath temperature enhances the contractility of the vas deferens in response to the release of neurotransmitters and to the application of exogenous noradrenaline via a postsynaptic action. In addition, via a presynaptic action, lowering the bath temperature depresses or delays the release of the noradrenergic transmitter from the sympathetic nerve terminals.     

失神经状态下胫骨骨折断端骨痂成骨的组织学变化

摘要背景：有研究表明神经系统在骨折的修复重建中发挥重要的作用,但其具体机制尚未阐明,中枢神经及神经生长因子对骨痂中成骨的影响至今少有报道。目的：观察失神经状态下胫骨骨折断端骨痂成骨的组织学变化。方法：用SD大鼠建立左胫骨闭合骨折髓内固定大鼠模型,建模成功后随机为3组,左胫骨骨折组;T10脊髓横断组：于左胫骨骨折后,横断切除T10段脊髓约0.3 cm,制成T10脊髓完全性损伤大鼠胫骨骨折模型;神经生长因子治疗组：建立T10脊髓完全性损伤大鼠胫骨骨折模型后,肌注神经生长因子。术后各组均用苏木精伊红染色法观察骨组织形态变化,骨钙素免疫组织化学法观察骨痂中成骨情况,透射电镜观察成骨细胞内微结构变化。结果与结论：T10脊髓横断组大鼠在失神经状态下,骨痂中成骨细胞的数量减少,骨钙素表达减少,成骨细胞内细胞器功能低下。神经生长因子治疗组的以上各组织学观察指标均较T10脊髓横断组均有不同程度的改善。结果显示,神经生长因子可部分改善脊髓横断失神经状态下的骨组织再生情况。