THSD7A在宫颈癌细胞侵袭、迁移等恶性生物学行为中的研究

目的 为了探索THSD7A对宫颈癌细胞的侵袭、转移以及凋亡等生物学行为的影响。方法 对宫颈癌C33a细胞和SiHa细胞培养及转染siTHSD7A基因,采用Western blotting和RT-qPCR方法在蛋白和基因水平对其验证。后应用Transwell小室实验、流式细胞术检测THSD7A沉默后对两种宫颈癌细胞的侵袭、转移、细胞周期及凋亡的改变。结果 与空白组和阴性对照组相比,沉默THSD7A的表达后,宫颈癌C33a细胞和SiHa细胞的侵袭、迁移能力明显下降（P＜0.01）;进入S期和G2/M期细胞数目减少,凋亡比例增高（P＜0.05）。结论 提示THSD7A可能作为一种致癌基因,在宫颈癌恶性生物学行为的发展过程中起到推进作用。     

血清壳多糖酶3样蛋白1、肝纤维化4项及AFP的检测在不同肝脏疾病中的应用价值探讨

目的探讨血清壳多糖酶3样蛋白1(CHI3L1)、肝纤维化4项及甲胎蛋白(AFP)在不同肝脏疾病中的应用价值。方法选取172例符合条件的慢性乙型肝炎病毒感染者,分为慢性乙型肝炎组、肝硬化组和肝癌组。收集同期健康体检者30例作为正常对照组。比较各组间指标的差异,采用受试者工作特征(ROC)曲线分析各指标鉴别诊断效能。采用Spearman秩相关分析各指标与疾病严重程度的相关性。结果血清CHI3L1及肝纤维化4项在肝硬化组和肝癌组明显高于慢性乙型肝炎组和正常对照组(P <0.05)。血清AFP浓度在肝癌组明显高于其他各组,上述差异均有统计学意义(P <0.01),其诊断肝癌的AUC为0.912。ROC曲线分析显示CHI3L1在肝硬化组和慢性乙型肝炎组,慢性乙型肝炎组和正常对照组之间的鉴别诊断效能较好,AUC分别为0.853和0.845。Spearman相关性分析显示肝脏疾病的严重程度与各血清学指标均呈正相关(P <0.05)。结论 CHI3L1、肝纤维化4项及AFP的检测可以辅助诊断慢性肝炎及其肝纤维化导致的肝硬化等不同肝脏疾病,值得在临床推广和进一步的研究。     

Use of a Clostridium perfringens vector to express high levels of SIV p27 protein for the development of an oral SIV vaccine

Clostridium perfringens is a normal bacterial flora of the small and large intestines of humans and other animals. The current study investigates the potential use of a noncytotoxic C. perfringens as an oral vaccine vehicle for expression and intestinal delivery of a large amount of SIV antigens. Here we report the construction of a recombinant C. perfringens vaccine vector expressing high levels of SIV p27 during sporulation. Following oral administration of this recombinant C. perfringens vaccine vector to mice, large amounts of intact p27 protein were detected in the terminal ileum where the majority of Peyer's Patches (PPs) are located. Furthermore, dendritic cells (DCs) beneath the mucosal surface in the PPs were able to capture SIV p27 antigen, when PPs were exposed to C. perfringens expressing SIV p27 antigen. In addition, uptake of C. perfringens was able to induce maturation of mouse DCs. These results support the potential use of C. perfringens as an oral SIV/HIV vaccine vector.     

PIAS3敲低对前列腺癌细胞增殖和凋亡的影响

目的研究PIAS3敲低对人前列腺癌细胞系DU145细胞增殖、细胞周期和凋亡的影响。方法构建PIAS3shRNA表达质粒pSilencer4.1/PIAS3,转染DU145细胞。MTT法检测细胞增殖,流式细胞仪分析细胞周期和凋亡。结果测序证实PIAS3shRNA表达质粒构建成功。转染后DU145细胞PIAS3蛋白表达明显下调。MTT分析显示PIAS3敲低促进细胞增殖,并存在剂量-效应关系。流式细胞术分析显示:PIAS3敲低后S期细胞比例增加,G0/G1期细胞比例减少,凋亡细胞比例减少。结论PIAS3敲低促进体外前列腺癌细胞增殖,抑制其凋亡。     

Extracellular K+ accumulations and synchronous GABA-mediated potentials evoked by 4-aminopyridine in the adult rat hippocampus

Transient changes in extracellular potassium concentration ([K⁺]₀) and field potentials were evoked by 4-aminopyridine (4-AP; 50–100 M) and recorded with ion-selective microelectrodes in CA1b, CA3b and dentate sectors of adult rat hippocampal slices. Long-lasting field potentials recurred at a frequency of 1/60 s (0.016±0.003 Hz) in association with increases in [K⁺]₀ which were largest and most sustained in the dendritic regions where afferent fibers terminate (dentate>CAl>CA3) and in the hilus. In stratum radiatum of CA1 or stratum moleculare of the dentate these fields had a peak amplitude of 1.4±0.29 mV, duration 8.3±1.6 s, and were accompanied by increases in [K⁺]₀ of 1.8±0.22 mM that lasted 32±5.5 s (n = 17 slices). Interictal epileptiform potentials, which were brief (<0.2 s) and more frequent at 1/3 s (0.30±0.02 Hz) were also present in CA1, CA3 and the hilus and associated with small increases in [K⁺]₀ (0.5 mM, duration 2 s). Interictal activity was blocked by 6-cyano-7-nitroquinoxalone-2,3-dione (CNQX; 5–20 M); the slow, less frequent potentials were resistant to both CNQX and dl-2amino-5-phosphonovaleric acid (APV; 50 M) and reversibly blocked (or attenuated by 80%) by bicuculline methiodide (BMI) (25–100 M). The BMI-sensitive potentials were also abolished by baclofen (100 M), an effect which was reversed by 2-OH-saclofen (100 M). Focal application of KCl or GABA in the absence of 4-AP evoked long-lasting field and [K⁺]₀ potentials which were similar to those evoked by 4-AP but more sustained. The proportional relationship between the amplitudes of field and K⁺ potentials with GABA closely resembled that observed for 4-AP; in contrast the slope of KCl-evoked responses was lower. Our results demonstrate that in the adult rat hippocampus 4-AP induces in many different regions accumulations of [K⁺]₀ in synchrony with the long-lasting field potentials, which are known to correspond to an intracellular long-lasting depolarization of the pyramidal cells. These changes are smaller than those which occur in the immature rat hippocampus — which may be related to differences in Na-K-ATPase and susceptibility to seizures. These events involve the activation of GABA_A receptors, are under the modulatory control of GABA_B receptors, and likely arise from the activity of GABAergic interneurons and/or afferent terminals. The long-lasting field potentials appear to reflect mainly the direct depolarizing actions of GABA and to a much more limited extent the associated accumulation of [K⁺]₀.     

多发性骨髓瘤负性相关基因DAZAP2蛋白高效表达和纯化

目的在前期的研究中，DAZAP2基因在多发性骨髓瘤（Multiple myeloma，MM）患者中表达明显下调，可能为MM的负性基因．须进一步获得DAZAP2蛋白，制备抗体，从而在蛋白水平上研究DAZAP2的表达、结构和功能．方法将原核重组质粒pQE30－DAZAP2转化大肠杆菌，阳性菌株经1 mmol／LIPTG诱导表达目的蛋白，再经过纯化和透析结果IPTG诱导4hr时，得到大量重组目的蛋白，占可溶蛋白的20％左右，经过Ni—NTA层析柱纯化。获得分子量为17kD的DAZAP2蛋白浓度为0．97mg／m1．结论DAZAP2蛋白获得高效表达，并且建立快速简便的纯化方法，目的蛋白可以用于下一步的实验．     

A simple quantitative protein A micro-immunoprecipitation method; assay of antibodies to the N and H antigens of poliovirus

Staphylococcus aureus (Cowan strain I) was used to absorb immune complexes from antiserum to poliovirus to which labeled N or H poliovirus antigens had been added, and the radioactivity in the pelleted organisms and in the supernatant was measured. Excellent agreement was obtained between values calculated separately from the pellet and supernatant readings, validating the use of supernatant measurements from a microtitration plate method.     

前S1蛋白在HBV指标中的重要意义

前S1蛋白是HBV衣壳蛋白的一部分，存在于大分子表面抗原蛋白之中，它是乙肝早期诊断的敏感指标，也反映了乙肝病毒的复制和乙肝病情的活动。本文研究显示，162例非乙肝病人血清中前S1蛋白全部阴性；311例乙肝患者中，前S1蛋白阳性率为38．9％，其中急性乙肝93．8％，慢性活动性肝炎51．2％，在乙肝二对半检测模式中，大三阳中该指标阳性率87．1％，小三阳为32．1％。以HBV DNA检测为标准，前S1蛋白的灵敏度为94．1％等，假阴性5．9％，总符合率93．9％。     

基于创新的四维密码方法分析共调控与蛋白互作关系

【摘要】目的 基于创新的四维密码子的方法挖掘共调控且互作的基因对，研究共调控与蛋白互作之间的关联趋势。方法 我们提出了创新的基于四维密码子的关联分析方法，并把此方法应用于与结肠癌相关的2000个疾病基因。结果 发现共调控的互作蛋白对比随机情况下显著；随着共调控强度的增强，蛋白间互作趋势越来越明显。结论 基于创新的四维密码子的关联分析方法，能够研究共调控与蛋白互作的关联趋势。     

Heymann肾炎致病原受体相关蛋白cDNA分子克隆及其mRNA在肾小球上皮细胞的表达

Ｈｅｙｍａｎｎ肾炎是一种用于人类膜性肾病的自身免疫病模型。ｇｐ３３０糖蛋白和４４ｋＤ受体相关蛋白（ＲＡＰ组成Ｈｅｙｍａｎｎ肾炎抗原复合物（ＨＮＡＣ），被证明是ＨＮ的主要致病原。本研究中，我们用ＲＴ－ＰＣＲ技术，克隆了ＲＡＰ基因，用Ｎｅｓｔｅｄ－ＰＣＲ技术克隆了ＲＡＰ羧基端１１８个氨基酸的基因，ＤＮＡ序列分析克隆基因与ＲＡＰ相同，原位杂交发现ＲＡＰｍＲＮＡ在肾小球上皮细胞表达。