伴肌动蛋白相关锚定蛋白（N-RAP）在颈椎后纵韧带骨化中的表达及意义

目的:从蛋白质及核酸水平研究伴肌动蛋白相关锚定蛋白(N-RAP)在颈椎后纵韧带骨化中的表达,探讨其存在的临床意义.方法:选择2006～2010年间行手术治疗的20例颈椎后纵韧带骨化症患者(OPLL组)及10例因外伤行颈椎手术患者(对照组)的颈椎后纵韧带标本.所有标本福尔马林固定、石蜡包埋,以N-RAP兔抗人多克隆抗体作为Ⅰ抗免疫组化SP法制片,DAB显色.选择染色满意的标本切片,观察表达N-RAP的细胞(棕黄染色)在两组样本中的数量及分布特点.5例骨化样本及5例对照样本冰浴匀浆、提取总RNA,行实时定量PCR检测.观测后纵韧带骨化症患者及对照病例后纵韧带组织中N-RAP mRNA的表达情况.结果:20例骨化韧带标本中,16例染色满意;10例对照组后纵韧带标本中,6例染色满意.免疫组化结果显示N-RAP表达于韧带组织中细胞的胞浆中,呈棕黄色染色.在OPLL组的后纵韧带组织中,可见多个黄染细胞及少量蓝色无N-RAP表达的细胞.而在对照组后纵韧带中,仅可见蓝色无N-RAP表达的细胞.实时定量PCR检测发现N-RAP在对照组标本中平均表达量为14.29±4.70,在OPLL组标本中为161.29±60.14,两组间差异有统计学意义(P＜0.05).结论:颈椎OPLL韧带组织中存在较高水平的N-RAP,其可能参与了韧带骨化的发生、发展过程.     

机械牵张应力对骨化颈椎后纵韧带成纤维细胞的影响

目的探讨机械牵张应力对体外培养的颈椎后纵韧带骨化症（OPLL）患者颈椎韧带成纤维细胞的影响。方法对2012年1月至2013年12月 OPLL 与颈椎外伤但无后纵韧带骨化（非OPLL）患者（各15例）行前路颈椎手术治疗,术中取韧带标本。采用组织块培养法进行细胞体外培养,免疫细胞化学及免疫荧光技术检测胞质波形蛋白。采用 Flexercell 4000细胞加载培养系统分别对两组患者第3代细胞进行机械牵张应力加载,逆转录-聚合酶链式反应（RT-PCR）方法检测两组成纤维细胞应力刺激前及刺激后12、24 h 成骨特异性指标骨钙素、碱性磷酸酶与Ⅰ型胶原 mRNA 表达。结果免疫细胞化学及免疫荧光检测显示胞质波形蛋白呈阳性表达。OPLL组后纵韧带成纤维细胞经机械牵张应力刺激12 h后,骨钙素、碱性磷酸酶及Ⅰ型胶原 mRNA 表达明显升高,应力刺激前后差异具有统计学意义;而非OPLL组应力刺激前后骨钙素、碱性磷酸酶及Ⅰ型胶原 mRNA 表达无明显变化。结论机械牵张应力可促使OPLL患者后纵韧带成纤维细胞骨钙素、碱性磷酸酶及Ⅰ型胶原mRNA表达增加,促进其骨化,其在OPLL进展中发挥重要作用。     

颈椎后纵韧带骨化椎间盘及后纵韧带应力的有限元分析

[目的]探讨颈椎后纵韧带骨化(OPLL)椎间盘及后纵韧带应力的变化。[方法]选择C4/5节段孤立型、混合型和节段型OPLL患者和正常人各1例行CT扫描,采用Mimics 14.0、Geomagic Studio 10.0和Hypermesh11.0软件对数据进行处理,建立有限元模型。采用Abaqus 6.12软件计算在前屈、后伸、旋转位载荷下椎间盘及后纵韧带的Von Mises应力。[结果]前屈和旋转载荷下,正常组C4/5、C5/6椎间盘应力显著小OPLL 3组(P0.05)。孤立型、连续混合型的C4/5、C5/6椎间盘应力显著小于节段型(P0.05)。前屈及旋转载荷下,正常组后纵韧带应力显著小于OPLL 3组(P0.05)。孤立型和混合型C4/5、C5/6节段后纵韧带应力显著大于节段模型(P0.05)。单开门成形术前与术后比较,C4/5、C5/6椎间盘及后纵韧带应力无显著改变(P0.05)。[结论]孤立型和混合型后纵韧带骨化的后纵韧带应力显著大于节段型,颈椎后路单开门不改变椎间盘与后纵韧带的受力状态。     

颈椎后纵韧带骨化症手术治疗的进展

颈椎后纵韧带起自第2颈椎,沿诸椎体后面抵于骶管。后纵韧带分为两层,浅层为一坚强韧带,自颅底垂直下行,在侧方延伸达椎间孔;深层呈齿状,椎体钩椎关节的关节囊一些纤维即始于此层[1] 。随着年龄增长,在众多因素作用下,后纵韧带组织中新生异位骨结构形成而逐渐发生骨化,导致椎管、椎间孔狭窄,压迫脊髓、神经根,临床上出现脊髓损害症状及神经根刺激症状,即为后纵韧带骨化症(ossi ficationofposteriorlongitudinalligament,OPLL )。OPLL在日本患者中较常见,所以又称为日本人病。OPLL患者通常有放射学表现,而无症状或只有轻微神经根、脊髓症…     

颈椎后纵韧带骨化症的自然史

颈椎后纵韧带骨化（ossification of the posterior longitudinal ligament,OPLL）是一种病因尚未明确的病理现象，表现为颈椎后纵韧带内异位骨的形成。1839年，Key首先报告了脊椎韧带的骨化现象。1960年，Tsukimoto根据尸体标本的解剖结果对颈椎OPLL进行了描述。1964年，Teray     

颈椎后纵韧带骨化症手术治疗进展

颈椎后纵韧带骨化(Ossification of the posterior longitudinal ligament of the cervical spine,OPLL)是一种原因未明的病理现象,表现为颈椎后纵韧带内异位骨形成。当骨化块压迫脊髓、神经根或血管引起临床症状时叫颈椎后纵韧带骨化症。近年来,有关手术治疗OPLL症的文献报道日益增多,手术方式亦不断改进,但每种方法均有其优劣之处及适应范围。本文就OPLL症的手术治疗进展作一综述。     

颈椎CT在OPLL临床诊治中的运用价值

后纵韧带骨化(ossification of the posterior longitudinal ligament,OPLL)在1838年被首次介绍,但直到19世纪60年代才被广泛报道[1]。日本学者依据患者颈椎平片上OPLL的不同特点将其分成四种类型:节段型、连续型、混合型以及局灶型[2]。颈椎平片在OPLL诊治中具有重要的运用价值,如Fujiyoshi等[3]把OPLL患者颈椎曲度及骨化大小合并一起并提出K-line的概念,认为K-line阴性的患者行后路手术术后改善不明显;而且颈椎平片因其单次费用[4]     

颈椎后纵韧带骨化症治疗策略研究进展

颈椎后纵韧带骨化症(ossification of the posterior longitudinal ligament,OPLL)为颈椎后纵韧带呈现进行性高度骨增生的异位骨化,导致增生骨化的韧带压迫脊髓,椎管体积减小[1、2],临床表现主要为脊髓病和或神经根病,伴有严重的神经病理学改变,导致肢体瘫痪和运动功能紊乱,降低生活质量。该病为OPLL的一种,并占OPLL总体发病率的70%[3、4],甚至在脊髓型颈椎病中的出现率高达25%,是引起脊髓型颈椎病的主要原因之一[5],应予高度重视。虽然早期的颈椎OPLL可保守治疗,但是在大多数颈椎OPLL患者中,病情是进展性的,甚至会导致毁灭性的神经系统并发症,如四肢轻瘫和四肢瘫痪,因此常需手术治疗[6、7]。现阶段临床中,治疗颈椎OPLL的手术方式可依据手术入路的选择分为前入路手术、后入路手术、后前入路联合手术。本研究将近10年发表于CNKI及Medline中有关颈椎OPLL的手术策略之适应证及优缺点进行如下综述。     

微RNA-218通过抑制RUNX2及Ⅰ型胶原基因表达延缓后纵韧带骨化

目的探讨微RNA-218(mi RNA-218)对后纵韧带骨化症(OPLL)患者原代后纵韧带细胞骨化的影响及其作用机制。方法原代培养5例OPLL患者及5例非OPLL者的韧带细胞,比较2组细胞miRNA-218的表达差异。利用agomir过表达或antagomir抑制OPLL患者韧带细胞中miRNA-218的表达水平后进行成骨诱导,通过检测茜素红染色水平、碱性磷酸酶活性及成骨相关基因的表达验证miRNA-218对韧带细胞骨化的作用。采用Target scan预测miRNA-218的靶基因,并采用双荧光素酶报告基因实验验证miRNA-218的靶向作用。结果 OPLL患者韧带细胞miRNA-218的表达水平低于非OPLL者,差异有统计学意义(P 0.05)。过表达miRNA-218后OPLL患者韧带细胞茜素红染色水平、碱性磷酸酶活性及成骨相关基因的表达均低于对照组;抑制miRNA-218后OPLL患者韧带细胞茜素红染色水平、碱性磷酸酶活性及成骨相关基因的表达均高于对照组;差异均有统计学意义(P 0.05)。Target scan预测miRNA-218靶基因可能为RUNX2和Ⅰ型胶原(COL1A1),双荧光素酶报告基因实验显示miRNA-218能降低RUNX2及COL1A1基因的荧光素酶活性水平。结论 miRNA-218能明显抑制原代后纵韧带细胞的骨化反应,其作用机制与抑制RUNX2和COL1A1基因表达相关。     

Deficiency of TBL1XR1 causes asthenozoospermia

Transducin (β)-like 1 X-linked receptor 1 (TBL1XR1) is an evolutionarily conserved protein related to spermatozoa. To clarify its role and mechanism of action in spermatozoa, qRT-PCR was used to analyse the expression of TBL1XR1 in human spermatozoa and mouse testes. The mice were established as an animal model by injecting the mice testes with small interfering RNA against TBL1XR1 or control siRNA. Our results indicated that deficiency of TBL1XR1 in mice reduced the motility of spermatozoa and disrupted the histone-to-protamine transition. We also found the decreased expression of TBL1XR1 in the spermatozoa of human patients with asthenozoospermia (AZ) compared with that in the spermatozoa of healthy males. Moreover, we carried out chromatin immunoprecipitation analyses and found that genes downstream of TBL1XR1 were related to sperm motility. Thus, TBL1XR1 might be related to sperm motility and might function through its downstream genes. Our data highlight the role of TBL1XR1 involved in spermatozoa and provide new molecular insights into the intricate systems required for male fertility.     

Alpha1-antitrypsin deficiency. 1: epidemiology of alpha1-antitrypsin deficiency

The protein and molecular characteristics of variants of the alpha1-antitrypsin (AAT) gene are described, and available data on the genetic epidemiology of AAT deficiency are presented.     

手术创伤中腹膜组织Sp1、COL1A1和TIMP-1的表达

目的　探讨人手术创伤腹膜组织中核转录因子Sp1激活 ,COL1A1和TIMP 1表达变化与腹膜纤维化之间的关系。方法　采用凝胶电泳迁移率改变分析法 (EMSA)检测手术创伤后不同时间的腹膜组织核转录因子Sp1的表达水平 ,WesternBlot检测COL1A1和TIMP 1蛋白表达 ,Masson染色观察腹膜组织中胶原纤维的变化。结果　Sp1在手术创伤后 0 .5h被活化 ,随着手术时间延长Sp1活性逐渐增强 ,至创伤后 4h时达高峰 ,同时创伤腹膜组织中的COL1A1和TIMP 1蛋白表达水平逐渐升高 ,存在差异显著性 (P <0 .0 1)。在手术创伤期内随手术时间的延长腹膜组织中胶原纤维增加。结论　核转录因子Sp1活化导致Ⅰ型胶原合成增加 ,细胞外基质降解减少 ,从而启动腹膜纤维化进程。     

Evaluation of the influence of polymorphic variants CYP1A1 2B, CYP1B1 2, CYP3A4 1B, GSTM1 0, and GSTT1 0 in prostate cancer

Background/objectiveGenetic polymorphisms in cytochrome P-450 (CYPs) and glutathione S-transferase (GSTs) genes can influence the appearance of tumors by the formation of new enzymes with altered activities. In the present study, 5 polymorphic variants were examined in 154 patients with prostate carcinoma and in 154 controls.Materials and methodsDNA analysis was carried out through PCR-based methods. The statistical methods used were odds ratio and confidence interval (95% CI), χ², Fisher, and Mann-Whitney.ResultsThe study showed absence of association for CYP1A1*2B, CYP1B1*2, GSTM1*0, and GSTT1*0. The statistical analysis implied a positive association of variant CYP3A4*1B for prostate cancer. The combined analysis of CYP1A1*2B, CYP1B1*2, and CYP3A4*1B genotypes showed positive association. The analysis of histopathologic parameters detected statistically significant differences for Gleason score and biochemistry recurrence risk. The presence of the GSTT1*0 genotype in red meat consumers increased the risk for this disease.ConclusionSome polymorphic variants analyzed can influence the development and the progression of prostate cancer.     

Genetic polymorphisms of CYP1A1, CYP2E1, GSTM1, and GSTT1 associated with head and neck cancer

BACKGROUND: Alcohol intake and tobacco smoke, in addition to other environmental and genetic factors, have been associated with head and neck cancer. We evaluated the role of metabolic enzyme polymorphisms on the risk of head and neck cancer in a hospital-based case-control study. METHODS: CYP1A1MspI, CYP2E1PstI, GSTM1, and GSTT1polymorphisms were evaluated in 103 histologically confirmed head and neck cancer cases and 102 controls by means of polymerase chain reaction-restriction fragment length polymorphism methods. RESULTS: GSTM1null increased the risk of head and neck cancer (odds ratio [OR], 2.2; 95% confidence interval [95% CI], 1.24-3.79), oral cancer (OR, 2.8; 95% CI, 1.28-5.98), and pharyngeal cancer (OR, 2.2; 95% CI, 1.08-4.63). CYP2E1PstI polymorphism indicated a risk for oral cancer (OR, 3.6; 95% CI, 1.29-11.56). The joint effect of GSTM1 null and CYP1A1 polymorphism increased the risk of head and neck cancer (OR, 2.4; 95% CI, 1.13-5.10). CONCLUSIONS: GSTM1 null alone or associated with CYP1A1 increased the risk of head and neck cancer; the CYP2E1PstI mutated allele increased the risk for only oral cancer.     

IgA肾病患者C1GALT1C1基因的体细胞突变筛查

目的 探讨IgA肾病（IgAN）患者β1,3半乳糖转移酶的分子伴侣Cosme编码基因C1GALT1C1基因体细胞突变情况。方法 27例IgA肾病患者及19例正常健康对照作为研究对象。提取研究对象外周血基因组DNA,扩增C1GALT1C1基因的编码区,采用PCR产物直接测序的方法进行突变筛查。然后,分离其中15例IgA肾病患者及7例健康男性对照的外周血B淋巴细胞,提取DNA。对C1GALT1C1基因编码区进行扩增,PCR产物进行克隆,各挑选平均8～10个克隆进行体细胞突变筛查。结果 46例个体全血基因组DNA的PCR扩增产物测序发现,2例患者及1例健康对照存在外显子T393A变异,次等位基因频率（MAF）为6．9%[SNP数据库（dbSNP）报告为9．5%]。B淋巴细胞DNA序列分析显示,在22例个体（15例IgA肾病患者,7例健康对照）送检的总共202个克隆中,未发现新的突变和多态性位点。结论 C1GALT1C1基因编码区T393A多态位点在本研究人群中为唯一发现的多态性位点,其次等位基因频率（MAF）较既往报道略低。本研究尚未发现IgA肾病患者B淋巴细胞存在体细胞突变。     

The HLA-DPB1--associated component of the IDDM1 and its relationship to the major loci HLA-DQB1, -DQA1, and -DRB1

The major histocompatibility complex (MHC) HLA region on chromosome 6p21 contains the major locus of type 1 diabetes (IDDM1). Common allelic variants at the class II HLA-DRB1, -DQA1, and -DQB1 loci account for the major part of IDDM1. Previous studies suggested that other MHC loci are likely to contribute to IDDM1, but determination of their relative contributions and identities is difficult because of strong linkage disequilibrium between MHC loci. One prime candidate is the polymorphic HLA-DPB1 locus, which (with the DPA1 locus) encodes the third class II antigen-presenting molecule. However, the results obtained in previous studies appear to be contradictory. Therefore, we have analyzed 408 white European families (200 from Sardinia and 208 from the U.K.) using a combination of association tests designed to directly compare the effect of DPB1 variation on the relative predisposition of DR-DQ haplotypes, taking into account linkage disequilibrium between DPB1 and the DRB1, DQA1, and DQB1 loci. In these populations, the overall contribution of DPB1 to IDDM1 is small. The main component of the DPB1 contribution to IDDM1 in these populations appears to be the protection associated with DPB1*0402 on DR4-negative haplotypes. We suggest that the HLA-DP molecule itself contributes to IDDM1.     

Impact of CYP1A1, GSTM1, and GSTT1 polymorphisms in overall and specific prostate cancer survival

ObjectivePrognostic biomarkers that distinguish between patients with good or poor outcome can be used to guide decisions of whom to treat and how aggressively. In this sense, several groups have proposed genetic polymorphisms as potential susceptibility and prognostic biomarkers; however, their validity has not been proven. Thus, the main goal of the present work was to investigate the potential role of single and combined CYP1A1, GSTM1, and GSTT1 genotypes as modifiers of cancer survival in Chilean patients with prostate cancer.Methods and materialsA total of 260 histologically confirmed patients were recruited from a voluntary screening, and genomic DNA was obtained from their blood samples for genotyping analyses to detect the CYP1A1*2A polymorphism and GSTM1 and GSTT1 deletions. The progression of illness and mortality were estimated with a median follow-up of 8.82 years. Adjusted estimated genotype risks were evaluated by hazard ratio and 95% CI using the Cox proportional model. In addition, the Kaplan-Meier survival method and log-rank test were used to evaluate patient survival with regard to genotype.ResultsThe 9-year overall and specific survival rates were 67.6% and 36.6% in the GSTT1null group, 67.6% and 58.7% in the GSTM1non-null group, 69.0% and 51.6% in the *1A/*2A group, 63.9% and 61.5% in the *2A/*2A group vs. 76.2% and 62.3% in the GSTT1non-null group, 82.3% and 50% in the GSTM1null group, and 83.7% and 56.3% in the *1A/*1A group, respectively. The hazard ratios and the Kaplan-Meier curve results demonstrate that the GSTM1non-null, GSTT1null, and CYP1A1*2A genotypes are significantly associated with mortality. Our study has two main limitations: a relatively small sample size and a low global mortality percentage (25.4%); thus, we need to continue the follow-up to confirm these findings.ConclusionsOur results suggest that the GSTM1non-null, GSTT1null, and CYP1A1*2A genotypes may be good prognosis markers, particularly in patients with high-risk tumors.