Genotype/phenotype analysis in Chinese laminin‐α2 deficient congenital muscular dystrophy patients

Laminin‐α2 deficient congenital muscular dystrophy (CMD) is an autosomal recessive disorder characterized by severe muscular dystrophy, which is typically associated with abnormal white matter. In this study, we assessed 43 CMD patients with typical white matter abnormality and laminin‐α2 deficiency (complete or partial) diagnosed by immunohistochemistry to determine the clinical and molecular genetic characteristics of laminin‐α2 deficient CMD. LAMA2 gene mutation analysis was performed by direct sequencing of genomic DNAs. Exonic deletion or duplication was identified by multiplex ligation‐dependent probe amplification (MLPA) and verified by high‐density oligonucleotide‐based CGH microarrays. Gene mutation analysis revealed 86 LAMA2 mutations (100%); 15 known and 37 novel. Among these mutations, 73.9% were nonsense, splice‐site or frameshift and 18.8% were deletions of one or more exons. Genetic characterization of affected families will be valuable in prenatal diagnosis of CMD in the Chinese population.     

Mimics软件模拟置钉在腰椎关节突重度退变椎弓根螺钉内固定中的应用

背景：目前腰椎椎弓根螺钉置入为脊柱外科非常成熟的一项技术,临床上有多种术中确定进钉点、进钉方向的方法。但腰椎关节突重度退变患者由于手术区域骨赘增生严重,顺利准确置钉有一定难度。 目的：观察腰椎关节突关节重度退变患者修复术前应用Mimics软件模拟置钉对椎弓根螺钉置入内固定效果的影响。 方法：将78例需要行腰椎椎弓根螺钉置入单节段内固定且治疗前CT、MR评估为腰椎关节突重度退变的患者纳入研究,随机分为两组,模拟置钉组39例,对照组39例,两组共置入312枚椎弓根螺钉。模拟置钉组术前对需要置钉的椎体使用Mimics软件行3D重建,并在拟置钉椎体模拟椎弓根螺钉,在3D影像上观察椎弓根螺钉进钉点与周围骨性标志的关系并测量进钉的角度和深度,术中参考观察和测量的结果进行椎弓根螺钉置入。对照组采用传统方式置入椎弓根螺钉。治疗后均行X射线、CT检查对椎弓根螺钉置入准确性进行评估。 结果与结论：模拟置钉组置入椎弓根螺钉后均未发生相关并发症,置钉后复查156枚螺钉中153枚位于椎弓根内,置钉正确率显著高于对照组(98.1%,88.5%,χ²=11.49,P < 0.05)。且模拟置钉组手术时间、出血量及患者放射线暴露时间少于对照组,差异均有显著性意义(P < 0.05)。提示需要行腰椎椎弓根螺钉置入内固定的患者中,腰椎关节突关节退变占相当大的比例,而关节突的退变使进钉点解剖标志难以辨别,给螺钉置入带来很大的难度。Mimics软件模拟置钉可辅助腰椎关节突重度退变患者修复术中椎弓根螺钉的正确置入,降低相关并发症,减少手术时间及出血量,减少患者放射线暴露时间。 中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程全文链接：     

肝门静脉海绵样变性模型大鼠组织抑制因子及基质金属蛋白酶表达 与周围血管新生

背景：目前国内外尚无有效治疗肝门静脉海绵样变性的策略,且其病因的基础研究报道较为鲜见。 目的：拟建立大鼠门静脉海绵样变性模型,检测基质金属蛋白酶2,9,基质金属蛋白酶组织抑制剂1,2在大鼠门静脉及其周围组织的表达及周围血管新生中的作用。 方法：SD大鼠80只随机分为3组。以门静脉部分缩窄法复制门静脉海绵样变性的大鼠模型,以21 G钝针头操作。模型组和假手术组分别设术后2,4,6周3个时间点,对照组即不做任何处理的正常SD大鼠(造影后取材)。各组分别于处理后不同时间点行门静脉造影,免疫组织化学检测血管内皮细胞标记物CD31观察门静脉周围血管变化,并使用实时荧光定量PCR和免疫组织化学法检测大鼠门静脉及其周围组织的基质金属蛋白酶2,9,基质金属蛋白酶组织抑制剂1,2 mRNA的含量和蛋白的表达。 结果与结论：门静脉造影及血管内皮细胞标记物CD31免疫组织化学显示,模型组门静脉周围新生血管明显增多。RT-PCR与免疫组织化学结果分析显示：对照组和假手术组基质金属蛋白酶2 mRNA及蛋白表达均较同期模型组低(P < 0.01,P < 0.05),基质金属蛋白酶9 mRNA及蛋白表达均较同期模型组低。模型组基质金属蛋白酶组织抑制剂1,2各个时间段的表达水平与对照组和假手术组相比差异无显著性意义(P > 0.05);模型组造模后第2周基质金属蛋白酶2/基质金属蛋白酶组织抑制剂2比值明显高于对照组和假手术组同期(P < 0.05)。结果提示,构建的门静脉海绵样变性的模型大鼠死亡率低,成模率高,且比较稳定。基质金属蛋白酶2,9表达升高以及基质金属蛋白酶2/基质金属蛋白酶组织抑制剂2的比值失衡,可能是大鼠门静脉海绵样变周围血管新生的分子机制之一。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

Prognostic significance of long non-coding RNA PCAT-1 expression in human hepatocellular carcinoma

Background: Long non-coding RNAs (lncRNAs) play widespread roles in gene regulation and cellular processes. However, the functional roles of lncRNAs in hepatocellular carcinoma (HCC) are not yet well elucidated. The aim of the present study was to measure the levels of lncRNA PCAT-1 expression in HCC and evaluate its clinical significance in the development and progression of HCC. Methods: We examined the expression of PCAT-1 in 117 HCC tissues and adjacent non-tumor tissues using quantitative real-time-PCR and analyzed its correlation with the clinical parameters. Results: Our data showed that PCAT-1 expression in HCC tissues was significantly increased compared with adjacent non-tumor tissues (P<0.05). Up-regulated expression of PCAT-1 was significantly associated with TNM stage and metastasis (P<0.05), but not other clinical parameters. Moreover, Kaplan-Meier survival analysis showed that a high expression level of PCAT-1 resulted in a significantly poor overall survival of HCC patients. The multivariate Cox regression analysis demonstrated that PCAT-1 expression level was an independent prognostic factor for the overall survival rate of HCC patients. Conclusions: Our data suggested that the increased expression of PCAT-1 was associated with advanced clinical parameters and poor overall survival of HCC patients, indicating that PCAT-1 up-regulation may serve as a novel biomarker of poor prognosis in HCC patients.     

Association of Genetic Variants of SIRT1 With Type 2 Diabetes Mellitus

SIRT1 has been demonstrated in nutrient-sensing and insulin-signaling pathways in in vivo and in vitro experiments, but there is minimal information concerning the association between gene polymorphisms of SIRT1 and type 2 diabetes mellitus (T2DM) in a Chinese Han population. Using case-control design, we recruited 310 unrelated T2DM patients from inpatients at Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, while 301 healthy controls were volunteers from the community for regular medical checkup. All participants were genotyped within the SIRT1 region. The following five SNPs rs10509291, rs12778366, rs10997870, rs10823112, and rs4746720 cover 100% of common genetic variations (minor allele frequency ≥ 0.05) within the SIRT1 gene (r ² ≥ 0.8). The genotypes of SIRT1 gene polymorphisms were analyzed by the Snapshot assay and DNA sequencing. The resulting data show that there was significant genetic differentiation in rs10823112 [p = 0.003; OR (95% CI) = 1.515 (1.152–1.994) for genotype], rs4746720 [p = 0.024; OR (95% CI) = 1.37 (1.037–1.674) for genotype], and rs10509291 [p = 0.002; OR (95% CI) = 1.551 (1.179–2.04) for genotype] between T2DM and control subjects. However, the result of rs4746720 was no longer significant after correction for multiple testing (p after Bonferroni correction = 0.12); the results of rs10509291and rs10823112 were still significantly different between the two groups (p after Bonferroni correction = 0.01 and 0.015, respectively). Linear regression analyses adjusting for age, gender, and body mass index (BMI) showed that HbA1c and HOMA-IR in subjects with rs10509291 AA genotype were higher than those with TT genotype in T2DM group (p = 0.045, p = 0.035, respectively). Together, our data show that genetic variation of the SIRT1 gene is related to insulin resistance and increase risk of T2DM in Chinese Han population. The risk allele A at SIRT1 rs10509291 was closely associated with T2DM, and subjects who were homozygous of the A allele were more likely to develop T2DM.Key words: Type 2 diabetes, Genetic variants, SIRT1, Chinese Han population     

纳米晶胶原基骨材料在腰椎后外侧植骨融合中的应用：安全性及效果

背景：腰椎后外侧植骨融合中需要进行植骨,但骨量常不够,需再取患者部分自体髂骨补充。 目的：探讨纳米晶胶原基骨材料在腰椎后外侧植骨融合中的应用安全性及效果。 方法：回顾性分析四川省眉山骨科医院2013年2月至2014年2月收治的52例腰椎退行性疾病患者的临床资料,均实施腰椎后外侧植骨融合治疗,术中所使用的植骨材料为患者自体骨颗粒混合纳米晶胶原基骨材料。 结果与结论：52例患者中,内固定后伤口一期愈合的有51例,患者局部皮肤均未出现皮温上升以及红肿、疼痛、流液等现象。患者内固定后功能和疼痛症状均得到不同程度的改善,对患者的内固定后植骨融合情况进行分析,可得内固定后3个月完全融合率达13%,内固定后6个月达54%,内固定后12个月达84%,且未出现假关节,随访12个月,所有患者均未出现螺钉松动或者移位等现象,对固定节段相临椎体情况进行检查,均未出现滑脱或者移位等情况。表明在腰椎后外侧植骨融合中应用纳米晶胶原基骨材料可以获得良好的效果,材料具有生物相容性及一定的安全性。 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

10月龄APP/PS1双转基因AD小鼠海马超微结构特征的观察

目的:观察APP/PS1双转基因小鼠海马内Aβ沉积的形态特征。方法:以10月龄雄性APP/PS1双转基因小鼠和C57BL/6小鼠为研究对象,采用Aβ免疫组化和透射电镜技术,观察海马部位Aβ的沉积情况及相关的病理特征。结果:在双转基因小鼠海马内存在致密性神经炎性斑块及营养障碍性突起,但脑微血管壁的Aβ沉积不甚明显;营养障碍性突起及神经细胞、微血管内皮细胞、周细胞内存在大量的自噬泡及次级溶酶体,提示该模型存在自噬溶酶体途径功能紊乱。结论:该鼠模拟了AD的老年斑及相关退行性变等病理改变,其中可能伴有细胞多种代谢的变化及血脑屏障功能的变化。从老年斑角度而言,该动物是成功的阿尔茨海默病动物模型,可用于AD的生化、病理研究及新治疗方法的探索。     

Abolishing Cell Wall Glycosylphosphatidylinositol-Anchored Proteins in Candida albicans Enhances Recognition by Host Dectin-1

Fungi can shield surface pathogen-associated molecular patterns (PAMPs) for evading host immune attack. The most common and opportunistic human pathogen, Candida albicans, can shield β-(1 3)-glucan on the cell wall, one of the major PAMPs, to avoid host phagocyte Dectin-1 recognition. The way to interfere in the shielding process for more effective antifungal defense is not well established. In this study, we found that deletion of the C. albicans GPI7 gene, which was responsible for adding ethanolaminephosphate to the second mannose in glycosylphosphatidylinositol (GPI) biosynthesis, could block the attachment of most GPI-anchored cell wall proteins (GPI-CWPs) to the cell wall and subsequently unmask the concealed β-(1,3)-glucan. Neutrophils could kill the uncloaked gpi7 mutant more efficiently with an augmented respiratory burst. The gpi7 mutant also stimulated Dectin-1-dependent immune responses of macrophages, including activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways and secretion of specific cytokines, such as tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and IL-12p40. Furthermore, the gpi7 null mutant could induce an enhanced inflammatory response through promoting significant recruitment of neutrophils and monocytes and could stimulate stronger Th1 and Th17 cell responses to fungal infections in vivo. These in vivo phenotypes also were Dectin-1 dependent. Thus, we assume that GPI-CWPs are involved in the immune mechanism of C. albicans escaping from host recognition by Dectin-1. Our studies also indicate that the blockage of GPI anchor synthesis is a strategy to inhibit C. albicans evading host recognition.     

产时手术在治疗出生缺陷儿及改善其预后中的价值

目的 探讨产时手术在治疗出生缺陷儿及改善其预后中的价值.方法 2008年8月至2009年11月,在中国医科大学附属盛京医院母胎医学中心通过彩色三维多普勒超声(彩超)筛查出有胎儿出生缺陷(淋巴管瘤和脐膨出各3例,膈疝和腹裂各2例,骶尾部畸胎瘤1例)的11例病例,通过MRI检查和胎儿染色体核型分析,明确为可行外科手术治疗的先天性疾病.11例产妇均行子宫下段剖宫产术及产时手术治疗出生缺陷儿,其中将胎儿取出宫外不断脐带的产时胎儿手术3例(膈疝修补术2例、骶尾部畸胎瘤切除术1例);子宫外产时处理(EXIT)后行产房外科手术6例(腹裂和巨型脐膨出修补术各2例、颈部和面部淋巴管瘤切除术各1例);分娩后立即行产房外科手术2例(脐膨出修补术和胸壁淋巴管瘤切除术各1例).对产妇进行常规的产后复查.随访出生缺陷儿手术后的生长发育及营养状态.结果 (1)手术结果:11例产妇均行剖宫产术娩出胎儿行产时手术治疗.其中3例实施产时胎儿手术治疗,手术时间平均为89 min;6例先对胎儿实施EXIT,在保持胎儿胎盘循环的情况下完成气管插管,EXIT平均时间为5.5 min,然后切断脐带,实施新生儿产房外科手术治疗;2例实施单纯产房外科手术,平均时间为37 min.除重度膈疝新生儿术后3.5 h死亡外,其他10例出生缺陷儿术后均存活至今.11例产妇剖宫产术及出生缺陷儿手术的母体平均失血量为275 ml,11例产妇术后均无发热及感染征象,子宫复旧良好,手术切口愈合良好.所有病例均未输血治疗,术后3～5 d出院.(2)随访结果:10例出生缺陷儿术后分别于1～18个月到我院儿科发育门诊随访,患儿体质量及身长等发育正常.其中,1例腹裂患儿术后1个月由于肠管旋转不良,喂养不耐受,体质量小于同龄婴儿,给予体位疗法治疗后,现喂养良好,体质量增加,术后4个月发育至正常水平.轻度膈疝患儿于术后2个月发生肺部感染,住院治疗2周后好转,患侧胸部X线片提示肺气胸比约1/4,术后6个月发现动脉导管未闭,复查胸部X线片,患侧肺叶几近全部扩张正常.1例巨型脐膨出患儿术前诊断为先天性轻度室间隔缺损,1年后复查心功能未受影响.骶尾部畸胎瘤患儿术后无自主排尿,10 d后排尿基本正常,1个月后排尿完全正常.结论 产时手术治疗可迅速终止疾病进一步发展,并明显改善出生缺陷儿的预后.