中国东北地区未经抗病毒治疗的HIV/AIDS患者HIV毒株的耐药基因变异研究

目的　研究我国东北地区未接受抗逆转录病毒治疗的HIV AIDS患者HIV毒株的逆转录酶和蛋白酶耐药变异情况 ,为开展大规模临床抗病毒药物治疗提供本底数据。方法　RT PCR和套式PCR扩增HIVpol区基因 ,双脱氧法测定逆转录酶和蛋白酶基因序列 ,与国际耐药数据库比对辨别耐药变异。结果　 (1) 5 3例患者毒株亚型分析结果 :B′亚型 4 7例 ,B′ C亚型 4例 ,A、B亚型各 1例 ;(2 )未发现逆转录酶和蛋白酶原发耐药变异存在 ,但发现存在逆转录酶抑制剂继发变异 :M4 1L(1.9% )、I6 3M (1.9% )、L74I (1.9% )、S6 8G (1.9% )、V75L (3.8% )、V10 6I (1.9% )、I135L T (5 .7% )、V179D (7.5 % )和V189I (1.9% ) ,无症状感染者RT继发耐药变异出现率为 11.8% ,而艾滋病患者为5 2 .6 % (P <0 .0 1)。存在大量蛋白酶耐药继发变异V77I (88.7% )、L6 3P (86 .8% )、E35D (81% )、A71V(2 4 .5 % )、R4 1K (15 .1% )、L10I (9.4 % )、R5 7K (9.4 % )、D6 0E (9.4 % )、N37D (5 .7% )、G16E (3.8% )、I15V (1.9% )、M36I (1.9% )、K5 5R (1.9% )和L89M (1.9% )。未发现明显的亚型特异性耐药变异。结论　在中国东北地区未接受抗逆转录病毒治疗的HIV AIDS患者中未发现毒株逆转录酶和蛋白酶耐药原发变异 ,但大量继发耐药变异的存在提     

215例急性和亚急性重型肝炎临床特征对比性分析

目的进一步了解急性重型和亚急性重型肝炎（简称急重和亚急重）患者的临床特征，以及它们之间的异同。方法收集和整理215例急重和亚急重住院患者的临床资料，使用X^2检验、t检验、回归分析等方法进行相关的统计学分析。结果①乙型肝炎病毒感染仍是急重和亚急重型肝炎的主要病因，均占30％以上。抗结核药物是药物性急重和亚急重肝炎的首要原因；②急重和亚急重患者肝性脑病发生率分别为78．13％和43．05％，差异有统计学意义（P〈0．001）；③急重患者的平均凝血酶原活动度低于20％，而亚急重患者平均凝血酶原活动度低于30％；④急重患者发生率前三位的并发症分别为肝性脑病、电解质紊乱及脑水肿；而亚急重则分别为腹水、电解质紊乱及肝性脑病；⑤急重和亚急重患者的病死率与病情最重时PT、WBC及中性粒细胞比例均呈正相关，而与PTA、TC均呈负相关；亚急重还与病情最重时TB、BLA及CRE呈正相关，与CHE、TG、PLT、ALB呈负相关。结论①急重和亚急重患者无论在好发年龄、肝性脑病发生率、肝性脑病出现时间，还是在凝血功能异常、预后与实验室指标等方面差异较大，属两个独立的疾病；②对于无肝性脑病的急重和亚急重患者，严重的凝血功能异常是一个重要的灵敏和特异性指标。     

47例骨髓穿刺干抽的临床和病理分析

分析４７例骨髓穿刺干抽的临床和病理改变，干抽占同期１３１９例骨穿的３．６％。根据骨髓细胞的密度和间质纤维增殖的程度，把干抽的原因分为４类：高细胞伴间质细胞增多者１９例（４２．２％）；间质细胞增多者１４例（３１．１％）；高细胞者８例（１７．８％）及低细胞者４例（８．９％）。主要疾病为：急性淋巴细胞白血病（２５．５％），急性非淋巴细胞白血病（１７．０％），慢性白血病（１４．９％），骨髓转移瘤（１０．６     

牵张延长下颌骨对二腹肌纤维型构成影响的研究

目的 :研究牵张延长下颌骨前后二腹肌的组织化学特征。方法 :应用肌球蛋白ATP酶 (pH9.4)染色法 ,观测了牵张前后 8只狗的二腹肌肌纤维类型 ,并用VIDAS图像分析仪测量其横截面积。结果 :ATP酶染色 ,二腹肌肌纤维分为Ⅰ型和Ⅱ型 ,Ⅰ型纤维数量较Ⅱ型纤维少。牵张延长下颌骨后 ,Ⅰ型纤维数量显著性增多 ,Ⅱ型纤维数量显著性减少。牵张成骨后 ,Ⅰ型纤维、、Ⅱ型纤维的横截面积与术前比较没有显著性差异。结论 :牵张延长下颌骨后 ,与牵张方向平行的二腹肌肌纤维型分布发生了与功能变化相适应的改变     

基于形态学的均值平移算法的神经元干细胞图像的自动分割

在神经元干细胞的图像分析中,准确快速的图像分割是干细胞分化增值自动追踪系统的基础。为了准确地分割低对比度的灰度神经元干细胞图像,本研究提出一种基于形态学运算和均值平移算法的神经元干细胞分割方法,称其为形态学的均值平移算法。此算法可以快速地获得任意形状细胞的图像,并且能检测到图像中多连接边缘不封闭的细胞。将此方法应用于神经元干细胞序列图像分割中并且将其与门限分割、水线分割和活动轮廓进行对比。实验结果证明,与其它的方法相比,此方法获得的细胞分割形状更接近于真实细胞的形状,并且能获得或接近于原始图像中准确的独立细胞数目。此方法可以获得正确的分割结果,为进一步图像处理奠定了良好的基础。     

中国HIV感染者细胞毒性淋巴细胞与CD4~+CD25~+调节性T细胞相关性研究

目的：对中国HIV感染者NK细胞、CD8^＋T细胞及胞内穿孔素、颗粒酶-B表达与CD4^＋CD25^＋Foxp3^＋调节性T细胞水平的相关性进行研究,探讨调节性T细胞在HIV感染中的作用机制。方法：选取73名HIV/AIDS患者（长期不进展组、无症状HIV组、AIDS组）,应用流式细胞仪胞内染色技术检测NK细胞、CD8^＋T细胞数量及胞内穿孔素、颗粒酶-B表达水平,分析其与CD4^＋CD25^＋Foxp3^＋调节性T细胞水平的相关性。结果：CD4^＋CD25^＋Foxp3^＋调节性T细胞百分率与NK细胞、CD8^＋T淋巴细胞数量呈明显负相关（P〈0.01）,与CD8^＋T细胞内穿孔素、颗粒酶-B表达百分率呈明显正相关（P〈0.05）,与NK细胞内穿孔素、颗粒酶-B表达水平绝对值负相关（P〈0.01）,与CD8＋T细胞内颗粒酶-B表达绝对值呈明显负相关（P〈0.01）,与CD8^＋T细胞内穿孔素表达绝对值无明显相关性（P〉0.05）。CD4^＋CD25^＋Foxp3^＋调节性T细胞绝对值与CD8^＋T细胞内穿孔素、颗粒酶-B表达百分率呈明显负相关（P〈0.05）。结论：中国HIV感染者细胞毒性淋巴细胞数量功能的变化与调节性T细胞显著相关,提示高水平的调节性T细胞可能与细胞毒性淋巴细胞耗竭相关,可能为导致疾病进展的原因之一。     

Nerve growth factor-induced hyperexcitability of rat sensory neuron in culture

Abnormal spontaneous firing of primary sensory neurons is considered to be a cause of neuropathic pain. However, pathogenic mechanisms of hyperexcitable sensory neurons in neuropathic model animals are unclear. We examined effects of chronic treatment of nerve growth factor (NGF), one of candidate mediators for the pathogenesis, on excitability of sensory neurons by voltage-clamped recording in a cell-attached configuration. From rat dorsal root ganglion (DRG) neurons cultured without NGF, only stable holding currents without spontaneous firing activity were recorded. On the other hand, more than 20% neurons cultured in the presence of NGF for more than 3 days showed spontaneous current spikes at frequencies between 0.1 and 5 Hz. Each spikes had an initial inward phase followed by the outward phase, resulted from spontaneous transient depolarization followed by transient hyperpolarization. These spontaneous spikes were abolished by tetrodotoxin, lidocaine and reduction of extracellular concentration of Na+ from 154 mM to 100 mM, in all-or-none fashion, suggesting that spontaneous current spikes reflected spontaneous action potentials. From these results, it became evident that DRG neurons of adult rats had a nature to respond to NGF and obtained the abnormal hyperexcitability to fire spontaneously.     

Lipofectin介导转移的外源性ADA基因在反复上感儿T淋巴细胞的表达

本文对４１例健康儿童和１７例反复上呼吸道感染患儿外周血淋巴细胞腺苷脱氨酶（ＡＤＡ）活性进行了检测。在此基础上筛选出２例反复上感伴ＡＤＡ活性低下患儿。在体外对这２例患儿的外周血Ｔ淋巴细胞进行培养后，以Ｌｉｐｏｆｅｃｔｉｎ（脂质体）介导的方法对其进行了外源性ＡＤＡ基因的基因转移。结果显示：２例患儿体外培养淋巴细胞ＡＤＡ活性较转基因前升高。同步进行的标志基因ｐＢＬａｃＺ的基因转移的检测结果也直观地证实了Ｌｉｐｏｆｅｃｔｉｎ介导的基因转移是成功的。该研究为ＡＤＡ－ＳＣＩＤ淋巴细胞基因治疗的研究提供了初步的体外实验资料。     

基于膝关节三维模型的髌股关节动力学特性研究

目的构建有效的膝关节三维模型以计算髌股关节动力学参数。方法基于CT图像处理和CAD技术, 重建包括股骨下端、胫骨上端及髌骨的膝关节的骨骼三维模型,并定义髌股关节运动起重要作用的股四头肌直线模型、韧带及其它软组织的非线性弹性纤维束模型,以股四头肌肉力为输入控制变量。结果构造出一个有效的膝关节三维模型。结论该模型可以有效的计算髌股关节动力学参数。     

Primary structure,functional characterization and developmental expression of the ascidian Kv4-class potassium channel

Ascidians belong to the primitive chordates and their larvae show symmetrical beating of the tail, which is reminiscent of the swimming pattern in primitive vertebrates. Since ascidian larva contains only a small number of neurons in their entire larval nervous system, they will potentially provide a simple model for the study of animal locomotion. In a step towards the goal of establishing the molecular basis underlying ascidian larval neurophysiology, we describe here a Kv4 class of voltage-gated potassium channel, TuKv4, from Halocynthia roretzi. Whole mount in situ hybridization indicates that TuKv4 is expressed in most of larval neurons including motor neurons. TuKv4-currents reconstituted in Xenopus oocytes show currents with similar properties to the lower-threshold A-type currents from cleavage-arrested ascidian blastomeres of neural lineage. However, the voltage-dependency of the steady-state inactivation and activation was shifted leftward by 20 mV, as compared with native A-type currents, suggesting that other components may be required to restore full function of the Kv4 channel. Unexpectedly, another isoform lacking C-terminal cytoplasmic region was also isolated. This truncated isoform did not lead to a functional current in Xenopus oocytes. RT-PCR analysis showed that the truncated form is transiently expressed during larval development, suggesting some developmental role for potassium channel expression.