河北省唐山地区汉族人HLA-DRB1*、DQB1*基因多态性分析

HL A是目前已知最复杂、最具多态性的遗传系统 ,在人类学、法医学、疾病关联、器官移植等领域有重要意义 ,具有显著的群体差异、种族差异和地区差异。我们应用聚合酶链反应 -序列特异性引物 (polymerase chain reaction- sequence specificprimer,PCR- SSP)方法随机对祖居唐山地区的 10 0名汉族健康人进行了 HL A- DRB1*、DQB1*等位基因分型 ,以了解唐山地区汉族人 HL A- DRB1*、DQB1*等位基因频率分布情况 ,为进一步与其它地方民族群体的研究结果做比较提供了有意义的资料。1　对象与方法1.1　对象　祖居唐山地区汉族无血缘关系…     

Interferon-β-1a

The presumed but as yet unspecified autoimmune-mediated basis for the pathogenesis of multiple sclerosis has led to attempts to modify the immune system of patients with this disease based on general and selective approaches. The rationale for using interferon-beta for the treatment of multiple sclerosis is based on its antiviral and complex immunoregulatory activities. Interferon-beta-1a (Avonex(R)) is a recombinant molecule which is indistinguishable from natural interferon-beta derived from human fibroblasts. Its precise mechanism of action is unknown, although effects on immune system processes which have been implicated in the pathogenesis of multiple sclerosis have been documented. T cell activation and migration into the CNS is a primary process in the pathogenesis of multiple sclerosis. In vitro and in vivo, interferon-beta-1a, compared with placebo or no treatment, significantly reduced expression of T cell surface activation markers, and significantly increased CNS levels of interleukin-10 - a potent inhibitor of cell-mediated immune responses. Pharmacokinetic studies indicate that intramuscular injection is the optimal route of administration for this formulation of interferon-beta-1a. In patients with relapsing-remitting multiple sclerosis who participated in a randomised double-blind trial, interferon-beta-1a 30mug (6 MIU) administered by intramuscular injection once weekly for 2 years (n = 158), compared with placebo (n = 143), significantly extended the time to onset of sustained neurological disability. Interferon-beta-1a also reduced the rate of disease relapse by approximately one-third compared with placebo, a finding which was supported by cranial magnetic resonance imaging (MRI) data showing significant reductions in lesion number and volume. Interferon-beta-1a was well tolerated, with influenza-like symptoms making up the majority of adverse reactions. The clinical significance of the beneficial effect of interferon-beta-1a on disease progression has been endorsed by the findings of a retrospective statistical analysis of the disability outcomes data obtained in the double-blind trial. In patients with relapsing multiple sclerosis, interferon-beta-1a is the only drug that has been demonstrated to significantly slow disease progression without excessive toxicity, in addition to significantly reducing the rate of relapse - measured clinically and by MRI. Notwithstanding the absence of long term tolerability data, and data from comparative trials with other agents, interferon-beta-1a represents a promising advance in drug therapy for relapsing multiple sclerosis.     

IL-1β转化酶研究进展

IL－1β转化酶（ICE）是一种ASP-特异性的半胱氨酸蛋白酶，它经过蛋白裂解作用而活化，通过形成（P20）2／（P10）2四聚体而发挥功能。ICE具有5种异构体，其活性可受到多种蛋白的调节。ICE与某些炎性疾病和恶性肿瘤的发生有关，并在Fas介导的凋亡中发挥一定作用。     

HLA-DRB1新等位基因DRB1*1610的发现

目的 对HLA新的等位基因HLA-DRB1*1610的分析.方法 采用商用DNA试剂盒抽提样本基因组DNA,利用HLA-DRB1组特异性引物PCR扩增先证者HLA-DRB1基因的第2外显子,PCR产物经割胶回收后进行测序分析.结果 先证者有两个HLA-DRB1等位基因,其中一个为HLA-DRB1*1202,另一个HLA-DRB1等位基因经BLAST验证为新的等位基因,新的等位基因序列已递交GenBank(DQ192647).与最接近的DRB1*160201等位基因序列相比,新的等位基因仅在第2外显子上有1个核苷酸不同,即第227位A→T,导致第47位氨基酸Tyr→Phe.结论 该等位基因为新的HLA-DRB1等位基因,被世界卫生组织HLA因子命名委员会正式命名为HLA-DRB1*1610.     

重组人TGF-β1对TGF-β1基因启动活性的影响

目的：探讨TGF-β1对人TGF-β1基因自身启动子活性的影响。方法：以人全血基因组DNA为模板，PER扩增获得TGF-β1基因转录起始位点上游5′端-1328～ 812bp和 227～ 812bp的片段，以此作为启动子与含氯霉素乙酰基转移酶(CAT)报告基因而不含启动子的pCAT-enhancer质粒构建成重组质粒，并将其转染至大鼠星状细胞株nFSC中，细胞在DMEM培养基中进行培养，用不同浓度的TGF-β1进行干涉，测定并比较细胞的CAT表达水平。结果：不同浓度的TGF-β1对大鼠的肝脏星状细胞的增殖具有抑制作用，而且这种抑制作用在实验用的浓度范围内具有剂量依赖关系；浓度为2．5ng/ml TGF-β1可以使转染phTGF0．585、phTGF1．120、phTGF1．423、phTGF1．680、phTGF2．140质粒的大鼠肝星状细胞的CAT活性分别增加2～5倍。结论：TGF-β1在大鼠nFSC细胞中对TGF-β1启动子活性具有自身促进作用，为进一步研究TGF-β1的调节机制提供了依据。     

DRB1*1454:中国汉族人群人类白细胞抗原-DRB1*14的常见等位基因

RB1*14等位基因的分布格局明显不同,DRB1*1405在两个群体中的分布比例比较接近,而DRB1*1454在南方汉族中的分布比例明显高于北方汉族.     

Decreased humoral antibody episodes of acute renal allograft rejection in recipients expressing the HLA-DQβ1*0202 allele

The present investigation was designed to show the effect of human leukocyte antigen (HLA) class II molecular allelic specificities in the recipient on the induction of humoral antibody rejection, identified by C4d peritubular capillary staining, as well as specific antibody identified by Luminex technology. Major histocompatibility complex (MHC) class II molecules are expressed on dendritic cells, macrophages, and B lymphocytes and they present antigenic peptides to CD4 positive T lymphocytes. Human renal peritubular and glomerular capillaries express class II MHC molecules upon activation. Expression of class II molecules on renal microvascular endothelial cells exposes them to possible interaction with specific circulating antibodies. We hypothesize that HLA-DQβ1*0202 expression in recipients decreases the likelihood of antibody-mediated renal allograft rejection. We found that 80% (=25) of DQ2 positive haplotype recipients failed to induce humoral antibody renal allograft rejection and 20% (n=25) of DQ2 positive haplotype recipients induced humoral antibody renal allograft rejection (p=0.008). By contrast, 48% (n=46) of DQ2 negative haplotype recipients failed to induce a humoral antibody component of renal allograft rejection and 52% (n=46) of DQ2 negative haplotype recipients induced humoral antibody-mediated renal allograft rejection. Our results suggest that recipients who express the DQβ1*0202 allele are less likely to induce a humoral antibody component of acute renal allograft rejection than are those expressing DQ1, DQ3, or DQ4 alleles. DQβ1*0202 allele expression in recipients could possibly be protective against acute humoral allograft rejection and might serve as a future criterion in recipient selection and in appropriate therapy for acute renal rejection episodes.     

ANXA1拮抗IL-1β导致的胰岛β细胞功能损伤的机制

目的探究ANXA1拮抗IL-1β引起的胰岛β细胞功能缺陷的作用及机制.方法将ANXA1基因插入pCMV-HA载体中,构建ANXA1过表达质粒,将其转染入小鼠胰岛β细胞系βtc-6细胞后,用IL-1β处理上述细胞,借助于MTT、葡萄糖刺激的胰岛素分泌实验(GSIS)等方法研究ANXA1对胰岛β细胞的保护作用.结果IL-1β导致胰岛β细胞ANXA1表达降低,同时出现胰岛β细胞存活率下降以及胰岛素合成和分泌量减少;而过表达ANXA1可拮抗上述现象的发生.结论ANXA1保护胰岛β细胞免受IL-1β造成的细胞凋亡增加与功能下降.     

Recombinant Interferon-β-1a

This review focuses on Rebif((R)), one of 2 available formulations of recombinant interferon-beta-1a, a molecule with the same molecular weight and primary structure as native interferon-beta. The product under review is intended for subcutaneous injection and contains 22 or 44microg of recombinant interferon-beta-1a. This molecule has the same antiviral, antiproliferative and immunomodulatory profile as native interferon-beta. Regulation of excessive immune responses in the inflamed lesions of patients with multiple sclerosis is thought to be important to its mode of action. In vivo studies indicate that the biological response to interferon-beta-1a is sustained with 3-times-weekly, in preference to once-weekly, administration. Subcutaneous interferon-beta-1a 22 and 44microg 3 times weekly for 2 years slowed sustained progression of disability (by 6.6 and 9.4 months; first quartile), decreased the mean number (by 27 and 33%) and severity of relapses, decreased the number of hospital visits and steroid courses, and decreased the acute activity [measured as the number of new or enlarging lesions seen with magnetic resonance imaging (MRI)] and burden (measured as the cumulative area or calculated volume of the lesions) of disease in patients with relapsing-remitting multiple sclerosis. All changes (except hospital visits: significant results were obtained with the higher dose only) were significant versus placebo with both doses. A recent study of once weekly interferon-beta-1a 22 or 44microg has confirmed the dose-dependency of the clinical and MRI effects. Patients with more severe disease appear to require the higher dosage regimen. Further studies of long term (>2 years) clinical efficacy, tolerability, and pharmacoeconomic aspects are required. Although injection site disorders and alterations in liver enzymes and lymphopenia are common, they rarely lead to withdrawal from treatment. As with other interferons, an influenza-like syndrome is often seen in patients receiving interferon-beta-1a. The sustained (over 2 years) presence of neutralising antibodies has been noted in 6 to 7% of patients; 16 to 18% of patients developed neutralising antibodies at some time during 2 years of treatment. This formulation is available as powder for reconstitution, or in liquid-prefilled syringes or an autoinjector device. CONCLUSIONS: Subcutaneous interferon-beta-1a 22 to 44microg 3 times weekly dose-dependently decreases the number and severity of relapses in patients with relapsing-remitting multiple sclerosis, slows the progression of disability, and decreases lesion activity and burden of disease. This formulation offers ease of dosage adjustment and convenience of administration, and is a valuable well-tolerated and effective addition to the choice of treatments for relapsing-remitting multiple sclerosis.     

PAI-1在TGF-β1基因和反义TGF-β1基因转染人肝癌细胞株中的表达

目的 :通过对人肝癌细胞株 (SMMC 772 1)转染TGF β1基因及反义TGF β1基因 ,观察该细胞PAI 1表达的变化。方法 :采用电穿孔法进行基因转染 ,并用Westernblot、Northernblot法加以鉴定后 ,分别应用Westernblot及Northernblot观察该细胞表达PAI 1mRNA的变化。结果 :过表达的TGF β1细胞克隆PAI 1mRNA表达均高于对照组 ,而低表达TGF β1者PAI 1mRNA表达低于对照组。结论 :TGF β1可能通过上调肝癌细胞PAI 1的表达 ,在肝癌浸润和转移过程中发挥重要作用。     

β-TCP材料改性研究进展及在骨缺损修复中的应用*

大段骨缺损再生修复是临床骨科面临的难题之一,利用组织工程材料修复大段骨缺损是最具前景的治疗方式。β-TCP材料由于具有与自体骨相似的无机成分,良好的生物相容性、可吸收降解以及骨传导等性能被广泛应用于骨科临床治疗。虽然β-TCP材料是一种最具潜力的骨替代材料,但是还存在诸如降解速率与新骨再生速率不匹配、脆性大力学强度不够、骨诱导能力差等缺陷,从而阻碍其临床应用。采用其他材料对β-TCP材料改性提高降解、力学及骨诱导等性能是目前研究热点。本文就β-TCP材料改性研究及在骨缺损再生修复中的应用作一综述。     

Cerebral small-vessel disease protein HTRA1 controls the amount of TGF-β1 via cleavage of proTGF-β1

Cerebral small-vessel disease is a common disorder in elderly populations; however, its molecular basis is not well understood. We recently demonstrated that mutations in the high-temperature requirement A (HTRA) serine peptidase 1 (HTRA1) gene cause a hereditary cerebral small-vessel disease, cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). HTRA1 belongs to the HTRA protein family, whose members have dual activities as chaperones and serine proteases and also repress transforming growth factor-β (TGF-β) family signaling. We demonstrated that CARASIL-associated mutant HTRA1s decrease protease activity and fail to decrease TGF-β family signaling. However, the precise molecular mechanism for decreasing the signaling remains unknown. Here we show that increased expression of ED-A fibronectin is limited to cerebral small arteries and is not observed in coronary, renal arterial or aortic walls in patients with CARASIL. Using a cell-mixing assay, we found that HTRA1 decreases TGF-β1 signaling triggered by proTGF-β1 in the intracellular space. HTRA1 binds and cleaves the pro-domain of proTGF-β1 in the endoplasmic reticulum (ER), and cleaved proTGF-β1 is degraded by ER-associated degradation. Consequently, the amount of mature TGF-β1 is reduced. These results establish a novel mechanism for regulating the amount of TGF-β1, specifically, the intracellular cleavage of proTGF-β1 in the ER.     

MDS患者外周血中TCR Vβ1-24-Dβ1 sjTRECs的分析

目的检测骨髓增生异常综合征（MDS）患者外周血中TCRVβ1-24-Dβ1 sjTRECs的存在情况。方法利用半巢式PCR扩增9例MDS患者外周血单个核细胞DNA中TCRβ链基因重排时形成的T细胞受体DNA删除环（Vβ1-24-Dβ1 sjTRECs），10例正常人外周血作为对照。结果所有TCRVβ1-24-Dβ1sjTRECs在正常人组外周血T细胞DNA中均有检出，除TCR Vβ12-Dβ1sjTRECs和TCRβ19-Dβ1sjTRECs外，其余TCRVβ-Dβ1 sjTRECs在MDS患者外周血T细胞DNA中均可检。大多数TCRVβ1-24-Dβ1sjTRECs在正常人组中的检出率较MDS患者高，但其中只有6种TCRVβ-Dβ1sjTRECs（分别是TCRVβ3、12、13、15、21、22-Dβ1sjTRECs）的检出率差异有统计学意义。结论本研究率先提供了MDS患者外周血T细胞中TCRVβ-Dβ1sjTRECs的存在情况。MDS患者TCRVβ-Dβ1sjTRECs出现频率低于正常人提示其胸腺近期输出功能存在部分缺陷情况。     

Rotavirus-neutralizing antibodies inhibit virus binding to integrins α2β1 and α4β1

Summary Rotavirus outer capsid proteins VP5^*, VP8^* and VP7 elicit neutralizing, protective antibodies. The α2β1 integrin is a cellular receptor for rotavirus that is bound by VP5^*. Some rotaviruses also recognize the α4β1 integrin. In this study, the effects of antibodies to rotavirus on virus binding to recombinant α2β1 and α4β1 expressed on K562 cells were determined. All neutralizing monoclonal antibodies to VP5^* tested (YO-2C2, 2G4, 1A10) and two to VP7 (RV-3:2, RV-4:2) inhibited rotavirus binding to α2β1. Rotavirus binding to α4β1 was reduced by 2G4 and neutralizing antibody F45:2, directed to VP7. However, a neutralizing antibody to VP8^* (RV-5:2) and one to VP7 (RV-3:1) did not affect rotavirus binding to these integrins. Virus-cell binding was unaffected by non-neutralizing antibody RVA to the rotavirus inner capsid protein VP6. The attachment of human rotavirus strain Wa to these integrins was inhibited by infection sera with neutralizing activity collected from two children hospitalised with severe rotavirus gastroenteritis. A negative reference serum did not affect rotavirus-cell attachment. As the binding of rotaviruses to α2β1 and α4β1 is inhibited by neutralizing antibodies to VP5^* and VP7, and serum from children with rotavirus disease, rotavirus recognition of these integrins may be important for host infection.     

IL-1β(白细胞介素1β)在大鼠外周血中的节律变化研究

研究证实白细胞介素 1β(interleukin- 1β,IL- 1β)在细胞免疫激活、抗肿瘤、减轻机体炎症反应中发挥重要作用。此外 ,还有导致厌食、体温升高、活动能力降低、激活下丘脑 -垂体 -肾上腺轴以及引起一氧化氮合酶含量增加等作用。而有关 IL - 1β在外周血中的节律报道甚少 ,仅 Zabel P等报道过健康人外周血中 IL- 1β具有节律性 ,但对常用作实验动物的大鼠其 IL - 1β在外周血中的节律却未见报道。鉴此 ,本研究就 IL - 1β在大鼠外周血中的节律作初步探讨 ,为进一步了解 IL - 1β的生理及病理学作用提供时间生物学资料。成年健康雄性 S…     

新的HLA-DRB1等位基因DRB1*1212的核苷酸序列分析

目的验证一个新的HLA等位基因HLA—DRB1＊1212的序列。方法采用盐析法抽提样本基因组DNA，利用HLA—DRB1组特异性引物PCR扩增先证者HLA—DRB1等位基因的第2外显子，PCR产物经割胶回收后进行测序分析，通过聚合酶链反应-序列特异性寡核苷酸探针方法验证测序发现突变点。结果先证者有两个HLA—DRB1等位基因，其中一个为HLA—DRB1＊090102，另一个HLA—DRB1等位基因，经BLAST验证为新的等位基因，新的等位基因序列已递交GenBank（AY899825）。与最接近的DRB1＊120101等位基因序列相比，新的等位基因仅在第2外显子上有1个核苷酸不同，即第199位A→C，导致第67位氨基酸Ile—Leu。结论该等化基因为新的HLA—DRB1等位基因，被世界卫生组织HLA因子命名委员会正式命名为HLA-DRB1＊1212。     

Hepatitis C virus-induced furin and thrombospondin-1 activate TGF-β1: role of TGF-β1 in HCV replication

In this study, we demonstrated the molecular mechanisms of TGF-β1 induction as well as proteolytic activation in HCV (JFH-1)-infected cells. Our studies showed the synthesis and secretion of TGF-β1 in HCV-infected cells which was reduced in the presence of Ca²⁺ chelators, an inhibitor of mitochondrial Ca²⁺ uptake, and antioxidants. We also showed that the expression of HCV NS proteins NS3/4A, and NS5A can induce TGF-β1 by cell-based luciferase assay. Furthermore, mutational analysis revealed that the functionally active protease domain of NS3 and N-terminus domain of NS5A are required for TGF-β1 activity. Using siRNA approach we demonstrated that HCV-induced furin and thrombospondin-1 (TSP-1) are involved in the proteolytic activation of TGF-β1. Our results also suggest that TGF-β1 positively regulates HCV RNA replication. Collectively, these observations provide insight into the mechanism of TGF-β1 activation, which likely manifest in liver fibrosis associated with hepatitis C infection.