HLA-DQB1 基因与中国湖北汉族人 SLE 及其自身抗体的相关性研究

目的：探讨ＨＬＡＤＱＢ１等位基因与系统性红斑狼疮（ＳＬＥ）及其自身抗体的相关性。方法：采用ＰＣＲ／ＳＳＰ技术对５２例中国湖北地区汉族ＳＬＥ患者及１４３例正常对照者进行了ＨＬＡＤＱＢ１基因分型，并采用免疫印迹技术检测患者血清中自身抗体。结果：ＳＬＥ患者ＤＱＢ１０６０８（９６２％，χ２＝１０５１，Ｐ＜０００５）基因频率显著升高，ＤＱＢ１０３０２（５７７％，ＲＲ＝０２６Ｐ＜００５，ＰＦ＝０１４）和ＤＱＢ１０５０１（１９２％，ＲＲ＝０１１，Ｐ＜００１，ＰＦ＝０１３）基因频率显著降低，与正常对照组比较，ＤＱＢ１０６０８在伴抗Ｓｍ（１０３４％，Ｐ＜０００５）、抗ＲＮＰ（１１５４％，Ｐ＜０００５）、抗ｄｓＤＮＡ（２２２２％，Ｐ＜０００５）抗体阳性的ＳＬＥ患者中频率显著升高。结论：ＤＱＢ１０６０８与ＳＬＥ关联，并分别与抗Ｓｍ、抗ＲＮＰ、抗ｄｓＤＮＡ抗体的产生有相关性。而ＤＱＢ１０３０２、ＤＱＢ１０５０１等位基因对ＳＬＥ可能具有保护性。     

肝炎后肝硬变肝损害与细胞免疫功能(英文)

目的研究肝炎后肝硬变（ＰＨＣ）患者的细胞免疫状态及其与肝功能损害的关系．方法５１例ＰＨＣ患者，包括ＣｈｉｌｄＰｕｐｈＡ级２０例、Ｂ例１８例、Ｃ级１３例和２２例健康对照者，外周血经用ＦｉｃｏｌＨｙｐａｑｕｅ梯度离心分离单个核细胞后，采用３ＨＴｄＲ掺入技术测定了淋巴细胞转化，ＩＬ２和ＮＫ细胞活性．结果在ＰＨＣ患者淋巴细胞转化指数（ＳＩ）、ＩＬ２活性（ＳＩ）和ＮＫ细胞活性（％）较对照组均明显降低（１８１±１３０ＶＳ３４９±２１７，Ｐ＜００１；８１±６０ＶＳ１３６±５８，Ｐ＜００１；４０３±２１７ＶＳ６１３±２０５，Ｐ＜００１）．免疫功能缺陷与ＣｈｉｌｄＰｕｐｈ分级有关，Ｃ级明显低于Ａ、Ｂ级（Ｐ＜００１），Ｂ级低于Ａ级（Ｐ＜００５）．结论ＰＨＣ患者存在细胞免疫功能缺陷，且与肝损害程度有关．     

糖尿病酮症酸中毒并肝损害临床分析

目的探讨导致糖尿病酮症（ＤＫ）及酮症酸中毒（ＤＫＡ）患者肝损害的相关因素．方法ＤＫ或ＤＫＡ患者９９例,其中ＡＬＴ及ＡＳＴ均异常升高１１例（Ａ组）,单项ＡＬＴ异常升高１３例（Ｂ组）,肝功能正常７５例（Ｃ组）,对以上各组患者的血二氧化碳结合力（ＣＯ２ＣＰ）、尿素氮（ＢＵＮ）、血糖（ＢＧ）和血浆渗透压（ＯＳＭ）进行了统计分析．结果Ａ,Ｂ两组患者的ＣＯ２ＣＰ明显低于Ｃ组（Ｐ＜００１,ｔ＝６３３和ｔ＝６４３）,而ＢＵＮ则明显升高（Ｐ＜００１,ｔ＝３６１,ＡｖｓＣ;Ｐ＜００１,ｔ＝４３５,ＢｖｓＣ）,Ａ组的ＢＧ（Ｐ＜００５,ｔ＝２８４）和血浆ＯＳＭ（Ｐ＜００５,ｔ＝３１０）水平也显著高于Ｃ组,而Ｂ组患者的ＢＧ及血浆ＯＳＭ与Ｃ组比较无差异;与Ｂ组相比,Ａ组患者的ＣＯ２ＣＰ明显降低（Ｐ＜００２,ｔ＝２７１）,ＢＧ（Ｐ＜００５,ｔ＝２８９）和血浆ＯＳＭ（Ｐ＜００５,ｔ＝２３６）明显升高．此外,Ⅰ型糖尿病患者血清转氨酶异常升高的发生率明显高于Ⅱ型糖尿病患者（Ｐ＜００５,χ２＝４３８）．结论酸中毒和脱水是导致糖尿病酮症及酮症酸中毒患者肝损害的重要因素,酸中毒及脱水程度与肝损害程度相关．     

单克隆抗体SC3A在胃癌及癌前病变的表达意义

目的研究单克隆抗体ＳＣ３Ａ在胃癌及癌前病变的表达意义．方法应用免疫组化ＡＢＣ法及粘液组化染色检测１０１例胃良恶性病变组织中ＳＣ３Ａ的表达．结果胃癌７１例中ＳＣ３Ａ阳性５７例（８０３％），但与癌组织类型、分化程度、转移及术后生存率无明显关系．ＳＣ３Ａ阳性率在酸性粘液（＋）组胃癌明显高于酸性粘液（－）组（９０２％对２００％，Ｐ＜００１），硫酸粘液（＋）组胃癌明显高于硫酸粘液（－）组（９１３％对６００％，Ｐ＜００１）．而且癌旁肠化硫酸粘液阳性率明显较良性病变伴肠化高（８８９％对３５３％，Ｐ＜００１）；硫酸粘液（＋）组肠化ＳＣ３Ａ阳性率明显高于硫酸粘液（－）组（６０９％对３１３％，Ｐ＜００５）．结论单克隆抗体ＳＣ３Ａ的表达对胃癌诊断及组织发生探讨有一定意义．     

胃癌组织P53蛋白和PCNA的表达意义

目的研究胃癌组织Ｐ５３蛋白和增殖细胞核抗原（ＰＣＮＡ）的表达意义．方法应用ＡＢＣ免疫组织化学技术检测７６例保存资料完整并经病理学证实的胃癌石蜡包埋组织和３０例正常胃粘膜Ｐ５３蛋白及ＰＣＮＡ的表达进行同步检测．结果正常胃粘膜无Ｐ５３蛋白表达，而６４４％（４９／７６）胃癌组织中Ｐ５３蛋白表达阳性，Ｐ５３蛋白表达阳性者其细胞增殖活性为８２１％±１７６％，明显高于Ｐ５３蛋白表达阴性组的６４２％±１４３％（Ｐ＜００１）．Ｐ５３蛋白阳性率和ＰＣＮＡ计数值与胃癌临床分期及复发呈正相关（Ｐ＜００１）．结论联合检测Ｐ５３蛋白和ＰＣＮＡ对胃癌诊断、分期及预后有重要价值．     

回生口服液对人IL-2水平及LAK细胞活性的影响

目的探讨回生口服液对人体ＩＬ２水平及ＬＡＫ细胞活性的影响．方法取健康献血者外周静脉血，分离出单个核细胞（ＰＢＭＣ），加入植物血凝素（ＰＨＡ），于９６孔微量培养板中，分别加入不同浓度的回生口服液，孵育２４ｈ，以ＣＴＬＬ２细胞作为检测细胞，用ＭＴＴ法测定ＩＬ２水平，同时设空白对照及阴性对照组；分离出的ＰＢＭＣ加入γＩＬ２，于２５ｍｌ培养瓶中共同孵育９６ｈ，制备ＬＡＫ细胞，然后将其与经不同浓度回生口服液处理后的ＳＭＭＣ７７２１，Ｈｅｌａ细胞株于９６孔微量培养板中共同孵育２０ｈ，用ＬＤＨ释放法测ＬＡＫ细胞活性，同时设空白对照．全部数据采用ｔ检验方法进行处理．结果回生口服液在浓度为１０ｍｇ／ｍｌ以上时能促进ＰＢＭＣ在ＰＨＡ作用下分泌ＩＬ２（Ｐ＜００１），在１ｍｇ／ｍｌ时即能拮抗１％（Ｐ＜００１）、５％（Ｐ＜００５）人血清ＩＬ２抑制物的作用，且都与药物浓度呈正相关，在高浓度（１００ｍｇ／ｍｌ）时还能增强ＬＡＫ细胞的活性（Ｐ＜００１）．结论回生口服液可提高人ＩＬ２水平，并增强ＬＡＫ细胞活性，以实现抗癌作用．     

134例老年肺结核患者痰结核菌耐药情况分析

目的探讨老年肺结核患者痰结核菌的耐药情况。方法采用ＢＡＣＴＥＣ法对１３４例老年痰菌阳性肺结核患者痰结核分支杆菌做耐药性检测。结果老年肺结核患者初始耐药率为３６８％，显著低于获得性耐药率的７５９％（Ｐ＜００１）；耐异烟肼（ＩＮＨ，Ｈ）、利福平（ＲＦＰ，Ｒ）、链霉素（ＳＭ，Ｓ）前者亦明显低于后者（Ｐ＜００１）。初治组耐２和３种药比例为６６％和５３％，与复治组的２２４％和２７７％相比差异有显著性（Ｐ＜００１）。至少耐异烟肼和利福平两种药的比例，初治组为９２％，显著低于复治组的５００％（Ｐ＜００１）。结论老年人肺结核耐药情况严重，尤其是获得性耐药和耐多药比例偏高，应引起足够重视     

紫外线照射自血回输对肺心病患者血液抗氧化能力的影响

为进一步探讨紫外线照射自血回输（ＡＵＶＩＢ）对肺心病急性加重期患者辅助治疗的机理，对４８例肺心病患者ＡＵＶＩＢ治疗前后血液中超氧化物歧化酶（ＳＯＤ）、丙二醛（ＭＤＡ）、谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）、过氧化氢酶（ＣＡＴ）活性进行了检测，并以正常人作对照。结果发现：肺心病患者ＳＯＤ水平虽高于正常对照组，但无显著差异（Ｐ＞００５），ＭＤＡ含量显著高于正常对照组（Ｐ＜００１），而ＧＳＨ－Ｐｘ与ＣＡＴ活性明显低于正常对照组（Ｐ＜００５）。经ＡＵＶＩＢ辅助治疗后，ＧＳＨ－Ｐｘ与ＣＡＴ活性显著升高（Ｐ＜００５），ＭＤＡ水平显著降低（Ｐ＜００５）。说明ＡＵＶＩＢ能提高肺心病患者血液抗氧化能力     

乙型和丙型肝炎病毒感染与肝细胞癌

为探讨乙型和丙型肝炎病毒（ＨＢＶ和ＨＣＶ）感染与肝细胞癌（肝癌）发生的关系，采用ＥＬＩＳＡ和聚合酶链反应（ＰＣＲ）对沈阳地区１１７例肝癌、１０７例肝硬化和４５例血液透析患者血清进行了ＨＢＶ和ＨＣＶ血清标志及ＨＢＶＤＮＡ和ＨＣＶＲＮＡ检测，并采用限制性片段长度多态性对其中７３例ＨＣＶＲＮＡ阳性血清进行了ＨＣＶ基因分型。结果，肝癌组ＨＢＶ感染率（６０７％）显著高于ＨＣＶ感染率（３３３％，Ｐ＜００１），肝硬化组ＨＢＶ感染率（４３９％）明显高于ＨＣＶ感染率（２９０％，Ｐ＜００５）；血液透析组ＨＢＶ和ＨＣＶ重叠感染率（２６７％）明显高于肝硬化组（１０３％，Ｐ＜００５）；各组均以ＨＣＶⅡ型为主（６５２％～８００％），ＨＣＶⅢ型次之（２００％～３１４％）。结果提示：沈阳地区肝癌的诱发因素仍以ＨＢＶ为主，血液透析患者ＨＢＶ和ＨＣＶ重叠感染的机会更大，ＨＣＶⅡ型感染在本地区ＨＣＶ相关性肝癌和肝硬化的发生中可能起主要作用。     

老年人睡眠呼吸暂停综合征与夜间心绞痛的关系

目的研究睡眠呼吸暂停综合征（ＳＡＳ）与夜间心绞痛的关系。方法对伴有或不伴有ＳＡＳ的老年冠心病患者９２例进行睡眠呼吸暂停和动态心电监测以及夜间心绞痛、无症状心肌缺血的观察。结果ＳＡＳ组夜间心绞痛１６例（３４８％），心肌缺血２７例（５８７％），心律失常４１例（８９１％），均较无ＳＡＳ组的４例（８７％）、８例（１７４％）和１６例（３４８％）明显增加（均为Ｐ＜００１）。夜间心绞痛和无症状心肌缺血发作次数与呼吸暂停次数、持续时间呈显著正相关（ｒ＝０４７５～０６１３，Ｐ＜００１），与血氧饱和度呈显著负相关（ｒ＝－０５２６，Ｐ＜００１）。ＳＡＳ患者经治疗夜间心绞痛和无症状心肌缺血显著改善（Ｐ＜００１）。结论老年人ＳＡＳ与夜间心绞痛有关。     

DEAD-box protein CYT-19 is activated by exposed helices in a group I intron RNA

DEAD-box proteins are nonprocessive RNA helicases and can function as RNA chaperones, but the mechanisms of their chaperone activity remain incompletely understood. The Neurospora crassa DEAD-box protein CYT-19 is a mitochondrial RNA chaperone that promotes group I intron splicing and has been shown to resolve misfolded group I intron structures, allowing them to refold. Building on previous results, here we use a series of tertiary contact mutants of the Tetrahymena group I intron ribozyme to demonstrate that the efficiency of CYT-19–mediated unfolding of the ribozyme is tightly linked to global RNA tertiary stability. Efficient unfolding of destabilized ribozyme variants is accompanied by increased ATPase activity of CYT-19, suggesting that destabilized ribozymes provide more productive interaction opportunities. The strongest ATPase stimulation occurs with a ribozyme that lacks all five tertiary contacts and does not form a compact structure, and small-angle X-ray scattering indicates that ATPase activity tracks with ribozyme compactness. Further, deletion of three helices that are prominently exposed in the folded structure decreases the ATPase stimulation by the folded ribozyme. Together, these results lead to a model in which CYT-19, and likely related DEAD-box proteins, rearranges complex RNA structures by preferentially interacting with and unwinding exposed RNA secondary structure. Importantly, this mechanism could bias DEAD-box proteins to act on misfolded RNAs and ribonucleoproteins, which are likely to be less compact and more dynamic than their native counterparts.DEAD-box proteins constitute the largest family of RNA helicases and function in all stages of RNA metabolism (1, 2). In vivo, many DEAD-box proteins have been implicated in assembly and conformational rearrangements of large structured RNAs and ribonucleoproteins (RNPs), including the ribosome, spliceosome, and self-splicing introns (3). Thus, it is important to establish how these proteins use their basic mechanisms of RNA binding and helix unwinding to interact with and remodel higher-order RNA structures.Structural and mechanistic studies have elucidated the basic steps of the ATPase cycle of DEAD-box proteins and have provided an understanding of the coupling between ATPase and duplex unwinding activities (4–11). The conserved helicase core consists of two flexibly linked RecA-like domains that contain at least 12 conserved motifs, including the D-E-A-D sequence in the ATP-binding motif II (3, 12). Binding of ATP and double-stranded RNA to domains 1 and 2, respectively, induces domain closure, which completes the formation of an ATPase active site at the domain interface and introduces steric clashes in the RNA binding site, leading to the displacement of one of the RNA strands (6, 7). ATP hydrolysis and inorganic phosphate release are then thought to regenerate the open enzyme conformation (4, 8, 13). Unlike conventional helicases, DEAD-box proteins have not been found to translocate, limiting the unwinding activity to short helices that can be disrupted in a single cycle of ATP binding and hydrolysis (4, 8, 9, 14–16). This mechanism is compatible with the physiological roles of DEAD-box proteins, because cellular RNAs rarely contain continuous base-paired regions that are longer than one or two helical turns.The interactions of DEAD-box proteins with structured RNAs have been extensively studied using two homologous proteins that function as general RNA chaperones: CYT-19 from Neurospora crassa and Mss116 from Saccharomyces cerevisiae. In vivo, CYT-19 is required for efficient splicing of several mitochondrial group I introns and can promote splicing of group I and group II introns in yeast mutants that lack functional Mss116 (17, 18). Both proteins have been shown to act as general RNA chaperones during group I and group II intron folding in vitro and are thought to act primarily by reversing misfolding of the intron RNAs, although additional mechanisms may be used for some substrates (17–23). Importantly, the chaperone activities of these and other DEAD-box proteins correlate with their ATP-dependent helix unwinding activities, suggesting that DEAD-box proteins function by lowering the energy barriers for transitions between alternative structures that involve disruption of base pairs (24, 25).In vitro studies using the group I intron ribozyme from Tetrahymena thermophila have been instrumental in probing the chaperone mechanism of CYT-19 (17, 26–28). This ∼400-nt RNA folds into a compact, globular structure composed of a conserved core and a series of peripheral elements that encircle the core by forming long-range tertiary contacts (Fig. 1) (29–31). Upon addition of Mg²⁺ ions, the majority of the ribozyme population becomes trapped in a long-lived misfolded conformation, which then slowly refolds to the native state (32). The misfolded intermediate is remarkably similar to the native ribozyme, forming a complete native network of secondary and tertiary interactions and a globally compact fold (33, 34). Despite these similarities, refolding to the native state requires extensive unfolding, including disruption of all five peripheral tertiary contacts and the core helix P3 (33, 35). To explain these results, a topological error has been proposed, wherein two single-stranded joining elements are crossed incorrectly in the core of the misfolded ribozyme, and transient disruption of the surrounding native structure is required for refolding (33, 35).Open in a separate windowFig. 1.The Tetrahymena group I intron ribozyme. (A) Secondary structure and mutations. Peripheral elements are colored and thick arrows mark the long-range peripheral tertiary contacts. Paired regions (P) and loops that were mutated in this study (L) are labeled based on group I intron nomenclature in ref. 31. The mutated regions are enclosed in dashed boxes and labeled in bold, with sequence substitutions indicated nearby. Sequences that were deleted to construct the helix truncation mutants (Fig. 6) are enclosed in gray dashed boxes and the replacement nucleotides are shown in gray italic font. (B) Tertiary structure model of the ribozyme (31). Peripheral elements (colored surface) and the locations of the long-range peripheral tertiary contacts (circles) are highlighted using the same color scheme as in A. The ribozyme core is shown in silver. The block arrows indicate the approximate positions of tertiary contacts not visible in each respective view of the ribozyme. The figures were prepared using PyMOL.Given the structural similarity between the native and misfolded ribozyme, it is interesting that CYT-19 can accelerate refolding of the misfolded intermediate by at least an order of magnitude without detectably unfolding the native ribozyme (26). Insights into this apparent preference for the misfolded ribozyme came from studies of two ribozyme mutants in which the tertiary structure was destabilized, making the stability of the native ribozyme comparable to that of the misfolded intermediate (28). CYT-19 unfolded the native and misfolded conformers of these mutants with comparable efficiencies, suggesting that the efficiency of chaperone-mediated unfolding depended on the stability of ribozyme tertiary structure. However, the mutations studied were concentrated in one region of the ribozyme, leaving open the possibility that CYT-19 recognizes local disruptions rather than global stability.Here we investigate the roles of RNA stability in CYT-19-mediated unfolding of the Tetrahymena ribozyme by using a series of ribozyme mutants with disruptions of each of the five peripheral tertiary contacts. We observe a strong correlation between CYT-19 activity and global stability of ribozyme tertiary structure. Further, we find that the RNA-dependent ATPase activity of CYT-19 depends on the accessibility of secondary structure in the ribozyme. Our results lead to a general model for recognition and remodeling of unstable or incorrectly folded RNAs by a DEAD-box protein.     

RNA degradosomes exist in vivo in Escherichia coli as multicomponent complexes associated with the cytoplasmic membrane via the N-terminal region of ribonuclease E

RNase E isolated from Escherichia coli is contained in a multicomponent "degradosome" complex with other proteins implicated in RNA decay. Earlier work has shown that the C-terminal region of RNase E is a scaffold for the binding of degradosome components and has identified specific RNase E segments necessary for its interaction with polynucleotide phosphorylase (PNPase), RhlB RNA helicase, and enolase. Here, we report electron microscopy studies that use immunogold labeling and freeze-fracture methods to show that degradosomes exist in vivo in E. coli as multicomponent structures that associate with the cytoplasmic membrane via the N-terminal region of RNase E. Whereas PNPase and enolase are present in E. coli in large excess relative to RNase E and therefore are detected in cells largely as molecules unlinked to the RNase E scaffold, immunogold labeling and biochemical analyses show that helicase is present in approximately equimolar amounts to RNase E at all cell growth stages. Our findings, which establish the existence and cellular location of RNase E-based degradosomes in vivo in E. coli, also suggest that RNA processing and decay may occur at specific sites within cells.     

慢性丙型肝炎患者甲状腺功能异常情况观察

目的 了解慢性丙型肝炎患者甲状腺疾病发生情况及其与肝功能、病毒复制、肝硬化发生及合并症的关系.方法 检测133例未接受干扰素治疗的慢性丙型肝炎患者血清中总三碘甲状腺原氨酸、总甲状腺素、游离三碘甲状腺原氨酸、游离甲状腺素和促甲状腺激素(thyroid stimulating hormone,TSH),并收集患者临床资料,...     

hfgl2凝血酶原酶短干扰RNA表达质粒的构建及其效应

目的 探索人fg12凝血酶原酶(hfg12)短干扰RNA(siRNA)在体外对hfg12凝血酶原酶基因表达的干预效应.方法 构建产生hfg12短发夹状RNA(shRNA)的载体p-hfg12shRNA,将其与hfg12-增强型绿色荧光蛋白(EGFP)融合基因表达质粒pEGFP-hfg12共转染到中国仓鼠卵母细胞(CHO细胞),转染48 h后,倒置荧光显微镜下观察EGFP在CHO细胞中的表达,并通过流式细胞仪检测荧光细胞阳性率.将P-hfg12shRNA和hfg12表达质粒pcDNA3.1-hfg12共转染到CHO细胞,通过实时荧光定量PCR和免疫组织化学检测hfg12基因转录和蛋白表达水平的变化.分为4组:共转染p-hfg12shRNA和pcDNA3.1-hfg12为干预组,共转染无关序列shRNA表达质粒和pcDNA3.1-hfg12为非相关干预组,仅转染pcDNA3.1-hfgl2为未干预组,不转染任何质粒为空白组.结果 在CHO细胞中,含p-hfg12shRNA的干预组较未干预组和非相关干预组的绿色荧光强度明显减弱,荧光细胞数明显减少;荧光表达阳性细胞率:干预组α为6.5%±0.2%,未干预组0t为40.1%±1.8%,两组比较,x<,2>=8.056,P<0.01,差异有统计学意义.抑制效率达85.5%.hfg12shRNA显著抑制了hfg12 mRNA和蛋白质的表达,干预组hfg12 mRNA水平为0.11±0.01,未干预组为0.84±0.06,两组比较,F=10.7,P<0.01,差异有统计学意义.结论 p-hfg12shRNA可在体外高效特异地抑制hfg12基因转录和蛋白的表达,为进一步的体内干扰实验奠定了基础.     

细胞内La蛋白对乙型肝炎病毒蛋白质表达的影响

目的 探讨细胞内La蛋白对HBV蛋白质表达的影响.方法 针对人La蛋白序列设计特异性的小分子干扰RNA(siRNA),通过体外转录的方法合成双链siRNA,并转染进入稳定表达HBV的HepG2.2.15细胞,实时荧光定量基因扩增技术测定HepG2.2.15细胞中的La蛋白mRNA变化水平;电化学发光分析技术检测HepG2.2.15细胞分泌的HBsAg和HBeAg变化情况.所得数据进行配对假设检验的统计学分析.结果 特异性siRNA干扰La蛋白mRNA的表达,使La蛋白在HepG2.2.15细胞中的表达降低;HepG2.2.15细胞分泌在培养上清液中的HBsAg和HBeAg表达显著下降,相关性分析显示,HBsAg、HBeAg变化水平与La蛋白mRNA变化水平呈正相关,相关系数分别为0.938和0.899.结论 特异性siRNA能够抑制细胞内La蛋白mRNA的表达,La蛋白可通过某种方式影响HBV蛋白质的表达.     

肝癌转移相关的微小RNAs在不同转移潜能肝癌细胞系的定量研究

目的 研究与转移密切相关的微小RNAs(miRNAs)在不同转移潜能肝癌细胞系的表达水平,探讨其在肿瘤转移过程中的生物学功能.方法 提取细胞系MHCC97H、MHCC97L、HepG2、L02的总RNA,通过反转录获得特异miRNA(miR-122a、miR-124a、miR-148a、miR-148b、miR-15a、miR-219、miR-30c、miR-338、miR-34a、Let-7g、miR-9)的cDNA,并应用TaqMan MGB探针法对其进行定量检测.采用AB17500系统软件V1.3.1采集Ct值,并使用miRNA内参基因RNU6B校正,相对定量计算公式RQ=2-△Ct,A Ct=CtmiRNAs-CtRNu6B.数据均经SPSS13.0统计软件包处理,采用t检验或非参数检验. 结果 转移相关的miRNAs(miR-124a除外)在MHCC97H与MHCC97L中表达,差异均有统计学意义.HepG2中miR-30c、miR-338、miR-34a和Let-g的表达水平明显高于L02,分别为miR-30c(8.41±0.40比6.82±0.29),miR-338(3.14±0.29比-2.36±0.32),miR-34a(0.71±0.40比-2.95±0.26),Let-7g(-4.07±0.55比-6.98±0.56),t值依次为2.948,12.656,7.484,3.684,P值均＜0.05,差异有统计学意义.而miR-148b,miR-9的表达则显著低于正常肝细胞,分别为miR-148b(1.96±0.51比3.76±0.28),miR-9(-4.38±0.86比-1.10±0.53),t值依次为-3.073,-3.324,P值均＜0.05,差异有统计学意义.miR-148家族中miR-48b在所测细胞系中的表达(5.46±1.21)均显著强于miR-148a的表达(1.29±0.35),Z=-5.097,P=3×10-7,差异有统计学意义.结论 可以利用肝癌细胞系列细胞平台进一步研究肝癌转移相关miRNAs在肿瘤转移过程中的生物学功能.     

C反应蛋白通过CD32影响人内皮祖细胞分泌白细胞介素8

目的研究C反应蛋白(CRP)对人外周血内皮祖细胞(EPCs)分泌白细胞介素8(IL-8)的影响。方法EPCs经培养鉴定后,分别加入浓度为0、5、10、25 mg/L的人重组CRP共同培养,依次设为对照组及低、中、高浓度组;另取EPCs分别加0、10 mg/L人重组CRP分为实验组和CRP组,先经抗CD32抗体(抗CD32组)、抗CD16抗体(抗CD16组)预处理后,再与人重组CRP 10 mg/L共同培养;以ELISA检测IL-8表达,以实时定量PCR检测CD32、CD16 mRNA的表达。结果与对照组比较,中、高浓度组显著抑制EPCs分泌IL 8(P<0.05);与实验组比较,CRP组IL-8降低;与CRP组比较,抗CD32组可拮抗CRP的作用(P<0.05),EPCs不表达CD16 rnRNA,表达CD32 mRNA。结论CRP抑制EPCs分泌IL-8,可能与CD32相关。     

补肝养髓法对老年性痴呆海马神经元RNA和Nissl体的定量研究

目的观察补肝养髓法对自发老年性痴呆模型海马神经元内RNA和Nissl体的影响.方法用跳台实验(step-downtest)从21月龄昆明种小鼠筛选出自发老年性痴呆(记忆障碍)鼠,随机分为空白对照组、西药对照组、补肝养髓小剂量组、补肝养髓大剂量组,另设老年学习记忆正常组(老年正常组).西药对照组给以喜得镇(Hydergine)0.6mg/kg,补肝养髓小、大剂量组分别予补肝养髓方6.80 g/kg及20.41g/kg,连续60 d,正常对照和痴呆对照组均灌以等量双蒸馏水(DW);脑组织冰冻切片,细胞化学方法显示大脑皮质和海马神经元内RNA和Nissl体,全自动显微图像分析系统定量检测相关脑区RNA和Nissl体含量.结果补肝养髓法能显著增高大脑皮质和海马锥体细胞的RNA和Nissl体含量,而且其作用呈现出一定的量效关系.结论补肝养髓方可明显改善自发老年性痴呆模型的学习记忆关键脑区海马的神经元结构,并显著改善其海马神经元的RNA代谢.