Connective tissue growth factor expressed in rat alveolar bone regeneration sites after tooth extraction

OBJECTIVE: To understand bone regeneration process after tooth extraction could be a clue to develop a new strategy for alveolar bone reconstruction. Recently, accumulated evidences support that connective tissue growth factor (CTGF) is implicated in tissue repair of many tissues. In this study, we investigated the spatial and temporal expression of CTGF in the rat tooth extraction sockets. DESIGN: Five weeks old wild type male rats (weighing 120 g) were used for this experiment. Expression of CTGF was determined by immunohistochemistry and in situ hybridization in the rat upper molar tooth extraction sockets at 2, 4, 7, 10 and 14 days after tooth extraction. RESULTS: CTGF was expressed strongly in the endothelial cells migrating into the granulation tissue at the bottom of the sockets during 4 days after tooth extraction. During the reparative process, no apparent chondrocyte-like cell appeared in the sockets, while osteoblast-like cells proliferated in the sockets with low CTGF expression at 7, 10, 14 days after extraction. As expected, no staining was observed with the preimmune rabbit IgG and CTGF sense probe. CTGF may play an important role in angiogenesis and granulation tissue formation specifically at early healing stage after tooth extraction to initiate alveolar bone repair. CONCLUSION: CTGF was expressed at early healing stage of the rat tooth extraction wound.     

口腔鳞癌组织CK19蛋白质与mRNA水平的关系及其临床意义

目的：探讨口腔鳞癌组织中细胞角蛋白19（CK19）的蛋白质表达与基因转录之间的关系及其临床意义。方法：采用免疫组织化学和荧光实时定量RT—PCR方法，对33例口腔鳞癌患者的癌组织及癌旁组织中CK19的蛋白质表达和mRNA水平进行检测，采用SPSS10.0软件包对数据进行配对t检验和相关分析。结果：口腔鳞癌组织中，CK19蛋白质和mRNA表达呈正相关，且均显著高于癌旁组织。癌组织CK19蛋白质表达阳性率为90，91％（30／33），mRNA水平是癌旁组织的2．21倍。CK19蛋白质和mRNA表达均与肿瘤的恶性程度显著相关。恶性程度越高，两者的表达水平也越高。结论：口腔鳞癌组织中CK19蛋白质和mRNA表达上升，并且与肿瘤恶性程度相关．CK19免疫组织化学和荧光实时定量RT—PCR技术在鉴别组织恶性程度上具有潜在的临床价值。     

釉基质蛋白体外诱导大鼠外胚间充质细胞骨桥素和骨钙素mRNA的表达

目的：了解体外培养环境中，大鼠外胚间充质细胞在两种不同形式釉基质蛋白(粗提型EMPs及纯化型EMD)诱导条件下，成骨分化标志骨桥素及骨钙素mRNA的表达水平。方法：酶消化法原代培养SD大鼠颌突外胚间充质细胞，在含釉基质蛋白(EMPs或EMD或EMPs EMD)的培养液中进行体外诱导培养，提取总RNA，采用RT-PCR法，以β-actin cDNA为内参照，检测骨桥素及骨钙素mRNA的表达水平。结果：1)各诱导实验组细胞均出现骨桥素mRNA的表达；2)仅EMPs EMD组出现骨钙素mRNA的表达。结论：外胚间充质细胞经釉基质蛋白特别是粗提型EMPs及纯化型EMD体外复合和诱导培养，可向成骨或成牙骨质方向分化，出现骨桥素及骨钙素mRNA的表达。     

小鼠釉丛蛋白基因不同mRNA剪接体的克隆

目的：克隆小鼠釉丛蛋白基因不同mRNA剪接体，分析小鼠釉丛蛋白的多态性。方法：采用异硫酸氰胍一步法从新生小鼠磨牙牙胚组织中提取总RNA，反转录成cDNA第一链，经PCR扩增出小鼠釉丛蛋白基因特异片段，播入pBS质粒，筛选阳性克隆，并经酶切和核苷酸序列分析鉴定阳性克隆。结果：克隆T1序列与已发表的完整釉从蛋白mRNA序列一致。克隆T2序列与完整釉丛蛋白mRNA序列比较，缺失序列的84-158的75个碱基片段。结论：小鼠釉从蛋白基因在转录mRNA时，可产生不同的剪接体。     

结肠癌组织中差异表达基因的筛选和鉴定

目的 筛选和克隆结肠癌和正常结肠组织差异表达的基因片段,为探讨结肠癌的发病机制提供线索。方法 应用基因差异显示技术（DD—PCR）,比较结肠癌和正常结肠组织基因表达的差异,对其中一条有明显差异的基因片段进行克隆、测序,测序结果提交GenBank数据库中进行同源性分析,并用半定量RT—PCR进行初步鉴定。结果 同源性分析表明,该片段与已知基因DDX32高度同源（99％）。RT—PCR结果显示,在结肠癌组织中该基因mRNA的表达水平显著高于正常结肠组织（P〈0．05）。结论 DD—PCR是筛选差异表达基因的有效手段;在结肠癌组织中DDX32基因的表达显著高于正常结肠组织,此结果为研究DDX32基因在结肠癌发病中的作用提供了线索。     

Lyophilization provides long-term stability for a lipid nanoparticle-formulated,nucleoside-modified mRNA vaccine

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A Multi-Targeting,Nucleoside-Modified mRNA Influenza Virus Vaccine Provides Broad Protection in Mice

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参芪排毒汤对宫颈癌组织人乳头瘤病毒E6/E7、P53和PRb基因蛋白表达的影响

目的:研究参芪排毒汤对宫颈癌组织人乳头瘤病毒(HPV)E6/E7 mRNA、肿瘤抑制基因(P53)、视网膜母细胞瘤蛋白(PRb)表达水平的影响与意义。方法:选取112例宫颈癌患者进入研究,按随机数字表划分成对照组与治疗组,两组均给予新辅助化疗联合腹腔镜宫颈癌根治术治疗,治疗组在新辅助化疗的同时给予参芪排毒汤口服; 术前评估两组临床疗效,统计不良反应发生情况,术中取癌旁健康组织与病灶病理组织样本检测HPV E6和E7 mRNA表达水平并计算E6/E7比值,检测样本中P53、PRb表达水平; 随访至少3年,比较两组患者的3年无病情进展生存时间(PFS)、总生存时间(OS),分析新辅助化疗后病理组织中HPV E6/E7、P53、PRb表达水平对PFS、OS的预测价值。结果:治疗组缓解率为83.93%,对照组为62.50%,治疗组缓解率高于对照组(P<0.05); 治疗组不良反应发生率为12.50%,对照组为17.86%,差异比较无统计学意义(P>0.05); 化疗后两组患者癌旁组织中HPV E6/E7比值比较,差异无统计学意义(P>0.05),病理组中HPV E6/E7表达高于治疗组(均P<0.05),两组癌旁与病理组织中P53、PRb比较,差异有统计学意义(均P>0.05); 化疗后宫颈癌组织中HPV E6/E7、P53、PRb表达水平对于患者无病情进展生存时间PFS、OS均具有较高预测价值(AUC>0.9,P<0.05); 治疗组患者PFS、OS均长于对照组(均P<0.05)。结论:在腹腔镜宫颈癌根治术前新辅助化疗中联合应用参芪排毒汤,能够提高术前疗效,抑制宫颈癌组织中HPV E6/E7表达、提升P53、PRb表达水平,从而起到延长患者无病情进展生存时间与总生存时间,改善预后的作用。     

Sex differences in circadian timing systems: Implications for disease

Virtually every eukaryotic cell has an endogenous circadian clock and a biological sex. These cell-based clocks have been conceptualized as oscillators whose phase can be reset by internal signals such as hormones, and external cues such as light. The present review highlights the inter-relationship between circadian clocks and sex differences. In mammals, the suprachiasmatic nucleus (SCN) serves as a master clock synchronizing the phase of clocks throughout the body. Gonadal steroid receptors are expressed in almost every site that receives direct SCN input. Here we review sex differences in the circadian timing system in the hypothalamic–pituitary–gonadal axis (HPG), the hypothalamic–adrenal–pituitary (HPA) axis, and sleep–arousal systems. We also point to ways in which disruption of circadian rhythms within these systems differs in the sexes and is associated with dysfunction and disease. Understanding sex differentiated circadian timing systems can lead to improved treatment strategies for these conditions.