不同来源的CD34^＋细胞归巢相关分子的表达

目的 研究不同来源CD34^＋细胞归巢相关分子（HRM）的表达情况。方法采用免疫磁珠法（MACS）分选不同来源的CD34^＋细胞，免疫荧光标记流式细胞仪测定HRM的表达。结果骨髓（BM）、动员后的外周血（mPB）及脐血（UCB）来源的CD34^＋细胞均高表达细胞黏附分子CD49d、CD49e、CD54、CD11a、CD62L、CD44、CD31。UCB来源的CD34^＋细胞表面表达的细胞黏附分子中，CD49e表达显著低于BM和mPB来源的CD34^＋细胞（P〈0．05），CD54表达显著低于mPB来源者（P〈0．05），CXCR4表达显著低于BM来源者（P〈0．05）。结论UCB来源的造血干／祖细胞归巢能力低可能是UCB移植造血重建延迟的原因之一。     

Dissociation of regional activity in the default mode network in first-episode,drug-naive major depressive disorder at rest

Background

Major depressive disorder (MDD) is associated with altered neural activity in the default mode network (DMN). In the present study, we used a fractional amplitude of low-frequency fluctuations (fALFF) approach to directly investigate the features of spontaneous brain activity of the DMN in patients with the first-episode, drug-naive MDD at rest.

Methods

Twenty-four first-episode, drug-naive patients with MDD and 24 age-, gender-, and education-matched healthy subjects participated in the study. The fALFF and independent component analysis (ICA) approaches were utilized to analyze the data.

Results

Patients with MDD exhibited a dissociation pattern of resting-state fALFF in the DMN, with increased fALFF in the left dorsal medial prefrontal cortex (MPFC) and decreased fALFF in the left parahippocampal gyrus (PHG). The increased fALFF values of the left dorsal MPFC were positively correlated to the Hamilton Rating Scale for Depression (HRSD) scores.

Conclusions

Our results first suggested that there was a dissociation pattern of resting-state fALFF in the DMN in patient with MDD, which highlighted the importance of the DMN in the pathogenesis of MDD.     

茶多酚对鼻咽癌细胞增殖、凋亡、迁移和侵袭的影响及机制的研究

目的:探索茶多酚对鼻咽癌细胞株HONE1增殖、凋亡、迁移和侵袭能力的影响,并研究相关的分子作用机制。方法:用不同浓度的茶多酚处理鼻咽癌HONE1细胞,分别用细胞计数法-8(CCK-8)法、荧光染色法、细胞划痕、Transwell侵袭实验、流式细胞术等方法,观察细胞生物学行为的变化;同时,检测茶多酚对鼻咽癌细胞中血管内皮生长因子(VEGF)表达的影响。结果:茶多酚能显著抑制鼻咽癌细胞HONE1的增殖,并呈浓度依赖性,而对正常鼻咽上皮细胞则无明显抑制作用;诱导HONE1细胞凋亡;茶多酚还有降低鼻咽癌细胞HONE1迁移和侵袭的能力,并明显减少鼻咽癌细胞内VEGF蛋白的表达。结论:茶多酚在体外能够抑制鼻咽癌细胞增殖,诱导细胞凋亡,并降低鼻咽癌细胞体外迁移和侵袭能力。     

Bioinformatic analysis of the pathogenic mechanism of talaromyces marneffei infection

Background:Talaromyces marneffei (T marneffei), known as a significant pathogen in patients with AIDS in Southeast Asia, is a dimorphic fungus, which can cause deadly systematic infection in immunocompromised hosts. What is more, the dimorphic phase transition has been reported as a conspicuous process linked with virulence. Interestingly, the yeast form was found in infected individuals, representing the pathogenic phase. However, few researches were found to study the mechanism of dimorphic transition. Thus, a diverse insight into the dimorphic switch mechanism, is urgently needed and we are the first one to research the mechanism of dimorphism.Methods:Firstly, we investigated the microarray of T. marneffei in the Gene Expression Omnibus database (GEO) for differentially expressed genes (DEGs). Then Database for Annotation, Visualization and Integrated Discovery (DAVID) v6.8 was employed to analyze the underlying enrichment and pathway in biological process of DEGs. Meanwhile, protein-protein interaction (PPI) network was constructed using STRING database. On the strength of the theory that similar amino acid sequences share similar structures, which play a decisive role on the function of protein, three dimensional structures of hub-genes were predicted to further investigate the likely function of hub-genes.Results:{"type":"entrez-geo","attrs":{"text":"GSE51109","term_id":"51109"}}GSE51109 was elected as the eligible series for the purpose of our research, including {"type":"entrez-geo","attrs":{"text":"GSM1238923","term_id":"1238923"}}GSM1238923 ({"type":"entrez-geo","attrs":{"text":"GSM23","term_id":"23"}}GSM23), {"type":"entrez-geo","attrs":{"text":"GSM1238924","term_id":"1238924"}}GSM1238924 ({"type":"entrez-geo","attrs":{"text":"GSM24","term_id":"24"}}GSM24), and {"type":"entrez-geo","attrs":{"text":"GSM1238925","term_id":"1238925"}}GSM1238925 ({"type":"entrez-geo","attrs":{"text":"GSM25","term_id":"25"}}GSM25). PMAA_012920, PMAA_028730, PMAA_068140, PMAA_092900, PMAA_032350 were the most remarkable genes in all of the three PPI networks, thus, were viewed as hub-genes. With regard to the three-dimensional construction, except that there was no significant prediction structure of PMAA_092900 with the criterion seq identify > 30%, GMQE: 0-1, QMEAN4: -4-0, the parallel templates for four structures were Crystal structure of Saccharomyces cerevesiae mitochondrial NADP(+)-dependent isocitrate dehydrogenase in complex with isocitrate, Organellar two-pore channels (TPCs), Yeast Isocitrate Dehydrogenase (Apo Form) and Crystal Structure Of ATP-Dependent Phosphoenolpyruvate Carboxykinase From Thermus thermophilus HB8 in order.Conclusion:The dimorphic transition of T. marneffei was viewed as a pathogenic factor and DEGs were observed. In-depth study of the function and pathway of DEGs revealed that PMAA_012920, PMAA_028730, PMAA_068140, PMAA_092900, PMAA_032350 were most likely acting as the hub-genes and were likely taking effect through regulating energy metabolism.     

α-玉米赤霉醇对小鼠骨髓间充质干细胞向成骨细胞分化过程中OPG和RANKL蛋白表达的影响

目的 探讨α-玉米赤霉醇(α-zearalanol,α-ZAL)对小鼠骨髓间充质干细胞(Bone marrow-derived mesenchymal stem cells, BMSCs)向成骨细胞增殖、分化的影响。方法 采用全骨髓培养差速贴壁法体外分离培养小鼠骨髓间充质干细胞,取第3代细胞分为空白对照组、雌二醇阳性对照组、低浓度α-ZAL组、中浓度α-ZAL组,高浓度α-ZAL组。镜下每天观察细胞形态变化,碱性磷酸酶染色鉴定成骨细胞纯度,MTT实验测定细胞增殖活性绘制细胞生长曲线,采用酶联免疫吸附法(ELISΑ)测定碱性磷酸酶(alkaline phosphatase,ALP)、骨形态发生蛋白2( Bone morphogenetic protein-2,BMP-2),运用蛋白免疫印迹法（Western blotting,WB）检测护钙素（Osteoprotegerin,OPG）和NF-KB受体活化配体（Receptor actiator of nuclear factor-KB ligand, RANKL）的蛋白水平表达变化。结果 不同浓度的α-玉米赤霉醇可显著的促进小鼠骨髓间充质干细胞向成骨细胞增殖、分化,增加细胞活性（P<0.05）,明显提高小鼠骨髓间充质干细胞向成骨细胞分化后ALP和BMP-2的表达（P<0.05）,并且显著的上调了成骨细胞内OPG/ RANKL的表达率（P<0.05）。结论 不同浓度的α-玉米赤霉醇可以促进骨髓间充质干细胞向成骨细胞增殖、分化,并且通过上调OPG/ RANKL的表达率抑制成骨细胞细胞向破骨细胞分化,有望成为临床骨折疏松症治疗的激素替代药物。     

促血管生成素-1基因重组腺病毒载体的构建及鉴定

目的构建携带小鼠促血管生成素-1(Ang-1)基因的重组腺病毒载体并鉴定。方法化学合成小鼠Ang-1基因经PCR扩增后亚克隆至穿梭质粒,重组质粒转化感受态细胞,所得转化子经PCR电泳分析并测序鉴定,携带外源基因的腺病毒穿梭质粒与携带腺病毒大部分基因组的辅助包装质粒共转染人胚肾细胞系HEK 293细胞,反复冻融获取重组腺病毒,终点稀释法测定重组腺病毒滴度,所得腺病毒转染293T细胞48 h收集培养上清,通过Western Blot分析Ang-1蛋白的表达。结果 PCR产物电泳分析可见大小约1.5 kbp的特征性条带,测序结果显示基因序列与Gen Bank提供的Ang-1序列一致,经HEK293细胞包装后腺病毒滴度为1×109pfu/ml,转染293T细胞后培养上清Western Blot分析可见大小58 k D特征性条带,即转染后能分泌正确大小的重组Ang-1蛋白。结论 Ang-1基因重组腺病毒载体可成功构建,为进一步研究Ang-1蛋白对造血干细胞动员及血管新生的作用提供实验依据。     

广西汉族IL-10启动子区基因多态性与HBV易感性及感染转归的关系

目的探讨白细胞介素-10(IL-10)基因启动子区-592A/C、-1082G/A位点单核苷酸多态性与乙型肝炎病毒(HBV)易感性及后临床转归之间的关系。方法 256例慢性HBV感染者,其中乙型肝炎肝硬化59例,慢性乙型肝炎151例,慢性HBV携带46例,以52例健康体检者作为对照组。采用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)技术,检测各组IL-10基因启动子区-592A/C、-1082G/A位点基因型及等位基因分布频率。结果肝硬化组、肝炎组、携带组与对照组的IL-10-592A/C位点AA、AC、CC基因型及A、C等位基因分布频率比较,差异均无统计学意义(P>0.05);IL-10-1082G/A位点GG、GA和AA基因型分布频率比较,差异有统计学意义(P<0.05),AA基因型频率分布依次为肝硬化组>肝炎组>携带组>对照组(P<0.05);各组IL-10-1082G/A位点G、A等位基因频率比较,差异有统计学意义(P<0.05),肝硬化组的A等位基因频率分布最高,其余依次为肝炎组、携带组及健康对照组(P<0.05)。不同HBV-DNA水平组的IL-10-592A/C、IL-10-1082G/A两位点基因型及等位基因分布频率比较,差异均无统计学意义(P>0.05)。结论IL-10-1082G/A位点AA基因型及A等位基因可能与广西汉族人群HBV易感性及其临床结局有关;而IL-10基因启动子区-592A/C、-1082G/A位点多态性可能与广西汉族人群感染HBV后的病毒复制水平无明显关系。     

高频超声检查在甲状腺结节良恶性诊断中的应用价值

目的 探讨高频超声检查对甲状腺良恶性结节诊断的临床应用价值. 方法 回顾分析经病理证实的223个甲状腺结节的高频超声表现,分析甲状腺肿块内部回声特点、肿块大小、形态、纵横比、边缘、有无钙化等指标. 结果 223个甲状腺结节中,病理诊断为甲状腺癌68个,良性结节155个;超声诊断为甲状腺癌73个,良性结节150 个;68 个甲状腺癌中,超声诊断恶性病变58个,阳性预测值为85.3%,155个甲状腺良性肿瘤中,超声诊断良性病变140个,阴性预测值为90.3%. 分别以结节内部呈低回声、砂粒样钙化为标准,诊断甲状腺癌的敏感性分别为88.2%和82.4%;以结节纵横比≥1或边界不清和(或)形态不规则为标准,诊断甲状腺癌的特异性较高,分别为95.4%和90.3%. 结论 高频超声检查对鉴别甲状腺结节良恶性具有很高的临床价值;结节纵横比≥1、内部呈低回声、砂粒样钙化、肿块边界不清和(或)形态不规则是诊断甲状腺恶性肿瘤的重指标,具有较高的诊断准确性.     

慢性肝损伤兔胆石症动物模型的构建

目的构建稳定、可行、可重复的兔慢性肝损伤胆石症模型。方法将90只新西兰大白兔随机分为空白对照组、肝纤维化组、肝纤维化胆石症组、肝硬化组和肝硬化胆石症组,空白对照组10只,其余各组各20只。空白对照组正常饮食,肝纤维化组、肝纤维化胆石症组予20%的CCl4-橄榄油溶液0.2ml/kg腹腔注射,肝硬化组、肝硬化胆石症组予25%的CCl4-橄榄油溶液0.3ml/kg腹腔注射,每4d1次,连续12周,进行肝组织病理学检查及Masson染色,计算结石形成率。结果肝纤维化组、肝硬化组、肝纤维化胆石症组和肝硬化胆石症组结石形成率分别为5%、10%、70%、70%,空白对照组、肝纤维化组与肝纤维化胆石症组成石率的三组间比较差异具有统计学意义(χ2=25.476,P<0.001),两两比较结果显示,空白对照组、肝纤维化组与肝纤维化胆石症组的比较差异均具有统计学意义(P1<0.001;P2<0.001);空白对照组、肝硬化组与肝硬化胆石症组成石率的三组间比较差异具有统计学意义(χ2=22.426,P<0.001),两两比较结果显示,空白对照组、肝硬化组与肝硬化胆石症组的比较差异均具有统计学意义(P1<0.001;P2<0.001)。结石主要为胆固醇结石,HE染色光镜下可见肝小叶结构遭到破坏,肝细胞肿大;Masson染色见纤维组织增多并变为蓝色,尤其在肝小叶汇管区较为明显,可见假小叶。结论通过用CCl4和高胆固醇、高热量致石饲料,能成功构建稳定、可行的兔慢性肝损伤胆石症模型,为慢性肝损伤兔胆石症实验动物模型的构建提供一定的借鉴和指导意义。     

超声内镜弹性成像对消化系肿瘤良恶性的鉴别诊断价值

目的初步探讨超声内镜实时组织弹性成像及弹性应变率比值(SR)对消化系肿瘤病灶良恶性的鉴别诊断价值。方法对26例患者32个消化系统实质性肿瘤病灶进行超声内镜实时弹性成像,采用5分法进行弹性成像评分;测量病灶与周围对照组织的SR,并进行组间比较。结果以评分≤2分的病灶为良性病灶,评分≥3分的病灶为恶性病灶,良恶性病灶组的超声弹性评分对比差异有显著性(P0.05),超声内镜弹性成像诊断消化系恶性肿瘤的敏感性87.0%,特异性88.9%,准确性87.5%。良性病灶组SR范围为0.01~7.33,中位SR为7.39;恶性病灶组SR范围为1.00~47.66,中位SR为20.07,良性病灶组的SR显著低于恶性病灶组的SR,两者的差异有显著性(P0.05)。结论消化系肿瘤良恶性组织弹性成像呈不同的图像特征,超声内镜实时组织弹性成像评分及SR有助于消化系肿瘤良恶性的鉴别诊断。EUS弹性成像不能代替FNA,但是可以协助FNA靶向活检,提高活检的阳性率。