Axonal protection by Nmnat3 overexpression with involvement of autophagy in optic nerve degeneration

Axonal degeneration often leads to the death of neuronal cell bodies. Previous studies demonstrated the crucial role of nicotinamide mononucleotide adenylyltransferase (Nmnat) 1, 2, and 3 in axonal protection. In this study, Nmnat3 immunoreactivity was observed inside axons in the optic nerve. Overexpression of Nmnat3 exerts axonal protection against tumor necrosis factor-induced and intraocular pressure (IOP) elevation-induced optic nerve degeneration. Immunoblot analysis showed that both p62 and microtubule-associated protein light chain 3 (LC3)-II were upregulated in the optic nerve after IOP elevation. Nmnat3 transfection decreased p62 and increased LC3-II in the optic nerve both with and without experimental glaucoma. Electron microscopy showed the existence of autophagic vacuoles in optic nerve axons in the glaucoma, glaucoma+Nmnat3 transfection, and glaucoma+rapamycin groups, although preserved myelin and microtubule structures were noted in the glaucoma+Nmnat3 transfection and glaucoma+rapamycin groups. The axonal-protective effect of Nmnat3 was inhibited by 3-methyladenine, whereas rapamycin exerted axonal protection after IOP elevation. We found that p62 was present in the mitochondria and confirmed substantial colocalization of mitochondrial Nmnat3 and p62 in starved retinal ganglion cell (RGC)-5 cells. Nmnat3 transfection decreased p62 and increased autophagic flux in RGC-5 cells. These results suggest that the axonal-protective effect of Nmnat3 may be involved in autophagy machinery, and that modulation of Nmnat3 and autophagy may lead to potential strategies against degenerative optic nerve disease.     

Apoptotic microtubules delimit an active caspase free area in the cellular cortex during the execution phase of apoptosis

Apoptotic microtubule network (AMN) is organized during apoptosis, forming a cortical structure beneath plasma membrane, which has an important role in preserving cell morphology and plasma membrane permeability. The aim of this study was to examine the role of AMN in maintaining plasma membrane integrity during the execution phase of apoptosis. We demonstrated in camptothecin-induced apoptosis in H460 cells that AMN delimits an active caspase free area beneath plasma membrane that permits the preservation of cellular cortex and transmembrane proteins. AMN depolymerization in apoptotic cells by a short exposure to colchicine allowed active caspases to reach the cellular cortex and cleave many key proteins involved in plasma membrane structural support, cell adhesion and ionic homeostasis. Cleavage of cellular cortex and plasma membrane proteins, such as α-spectrin, paxilin, focal adhesion kinase (FAK), E-cadherin and integrin subunit β4 was associated with cell collapse and cell detachment. Otherwise, cleavage-mediated inactivation of calcium ATPase pump (PMCA-4) and Na⁺/Ca²⁺ exchanger (NCX) involved in cell calcium extrusion resulted in calcium overload. Furthermore, cleavage of Na⁺/K⁺ pump subunit β was associated with altered sodium homeostasis. Cleavage of cell cortex and plasma membrane proteins in apoptotic cells after AMN depolymerization increased plasma permeability, ionic imbalance and bioenergetic collapse, leading apoptotic cells to secondary necrosis. The essential role of caspase-mediated cleavage in this process was demonstrated because the concomitant addition of colchicine that induces AMN depolymerization and the pan-caspase inhibitor z-VAD avoided the cleavage of cortical and plasma membrane proteins and prevented apoptotic cells to undergo secondary necrosis. Furthermore, the presence of AMN was also critical for proper phosphatidylserine externalization and apoptotic cell clearance by macrophages. These results indicate that AMN is essential to preserve an active caspase free area in the cellular cortex of apoptotic cells that allows plasma membrane integrity during the execution phase of apoptosis.     

对比不同类型干细胞对于缺血性脑卒中治疗的研究进展

成人中枢神经系统存在着一定量的神经干细胞,其具有两大关键能力;自我更新和多向分化潜能。缺血性脑卒中是一种由于由脑血流的缺失或减少引起的脑动脉闭塞,进而导致脑组织梗死的脑血管疾病。虽然对于脑损伤的药物治疗已经取得了一定的成果,但目前以干细胞为基础的治疗方法仍成为了研究热点。无论是内源性神经干细胞还是外源性神经干细胞移植均可在脑损伤后向远端损伤区迁移并分化成新的神经细胞,从而在中枢神经系统疾病尤其是脑梗死后进行组织修复和功能恢复。因此在这篇综述中,我们主要探讨不同类型的干细胞对脑梗死介导的脑损伤的应用潜能,对比不同类型干细胞对缺血性脑卒中的治疗优缺点。     

介入治疗成人非创伤性股骨头缺血性坏死的疗效分析

目的：分析介入治疗对于成人非创伤性股骨头缺血性坏死的疗效。方法：采用介入方法治疗95例共117髋非创伤性股骨头缺血性坏死。对比介入治疗前后DSA造影分型结果及血供异常例数。结果：117患髋中,Ia型的31病髋中,11髋（35．48％）介入治疗后动脉主干再通;Ib型的27髋中,24髋（88．89％）介入治疗后分支再通;II型的11髋中,5髋（45．45％）实质期股骨头缺损面积缩小;III型的10髋中,8髋（80．00％）静脉期见股骨头浓密染色明显减轻;Ⅳ型的33髋中,28髋（84．85％）可见动脉主干再通、分支增粗、实质期股骨头缺损面积缩小和静脉期股骨头染色减轻等。治疗前117病髋异常率为95．73％。经过介入治疗后,异常率为30．77％。治疗后异常率明显低于治疗前异常率（P〈0．01）。结论：介入治疗可有效改善非创伤性股骨头缺血性坏死血供异常,增加股骨头血供,具有操作简单、创伤小、疗效确切等优点。关键阗：股骨头缺血性坏死;介入治疗;数字减影血管造影     

慢性牙周炎患者龈沟液中IL-8 和TNF-alpha水平变化及临床意义

目的：探讨慢性牙周炎患者龈沟液中白细胞介素-8（IL-8）和肿瘤坏死因子-α（TNF-α）水平变化及临床意义,为临床诊疗提 供参考。方法：选择2013 年12 月到2015 年12 月我院收治的60 例慢性牙周炎患者为实验组,选择同期60 例牙周健康者为对照 组。对照组对象于体检时、实验组患者于牙周常规治疗前后收集龈沟液并记录牙龈指数（GI）、牙龈沟出血指数（SBI）、菌斑指数 （PLI）、牙周袋探诊深度（PD）、临床附着丧失（CAL）等牙周临床指标。测量并比较两组对象龈沟液中IL-8 和TNF-α水平。结果：实 验组患者治疗前GI、SBI、PLI、PD 及CAL 等牙周临床指标均明显高于对照组,差异具有统计学意义（P<0.05）;治疗后,实验组患 者各牙周临床指标明显低于治疗前,差异有统计学意义（P<0.05）,但与对照组比较差异无统计学意义（P>0.05）。实验组患者治疗 前龈沟液中IL-8 和TNF-α水平均明显高于对照组,差异有统计学意义（P<0.05）;治疗后,实验组患者龈沟液中IL-8 和TNF-alpha水 平均明显低于治疗前,差异有统计学意义（P<0.05）。实验组患者治疗前龈沟液中IL-8 水平与牙周临床指标PD 呈正相关性（r=0. 495,P=0.027）,TNF-α水平与牙周临床指标SBI、PD 呈正相关性（r=0.512,0.673;P=0.019,0.012）。结论：慢性牙周炎患者龈沟液中 IL-8 和TNF-α具有较高水平,两者可能与慢性牙周炎的发生发展有关,对于临床诊断慢性牙周炎具有积极的临床意义。     

生长因子颗粒蛋白前体和肿瘤坏死因子受体基因启动子区改变与阿尔茨 海默病的相关性研究

目的:探讨生长因子颗粒蛋白前体(PGRN)、肿瘤坏死因子受体(TNFR)基因启动子区改变以及全基因组DNA甲基化与阿尔茨海默病的相关性。方法:收集阿尔茨海默病患者血液样本80例以及健康对照血液样本80例,PCR扩增PGRN和TNFR基因启动子区并进行测序,观察两组间的单核苷酸多态性位点是否有差异。同时,用甲基化特异性PCR法检测启动子区DNA甲基化情况以及用ELISA法检测全基因组DNA甲基化水平。结果:在TNFR基因启动子区域发现阿尔茨海默病和对照组之间在多态性位点rs4149570和rs4149569有显著性差异(P0.001和P=0.033)。阿尔茨海默病患者全基因组甲基化水平为(0.79±0.29)%,显著低于对照组的(1.00±0.36)%(P0.001)。结论:TNFR基因多态性位点rs4149570和rs4149569的变异可能与阿尔茨海默病相关,全基因组甲基化水平降低可能与阿尔茨海默病相关。     

关节镜治疗髋关节疾病的临床研究

目的:研究髋关节镜治疗髋关节疾病的方法、疗效及适应证,探讨髋关节镜在髋关节疾病中的临床价值。方法:研究对象为86例有明显髋关节疼痛的患者,包括股骨头坏死(osteonecrosis,ON)(43%)、盂唇损伤(20%)、退行性关节病变(degenerative joint disease,DJD)(10%)、股骨头骨骺缺血性坏死(Legg-Calve'-Perthes,LCP)(10%)、髋关节游离体(10%)、髋关节疼痛(100%)、机械性损伤(78%)、运动损伤(56%)。对患者采用仰卧位进行髋关节镜检查,使用牵引床,300或700,前外侧入口。观察不同疾病在治疗后的预后结果。结果:所有患者均无并发症,平均随访时间30个月,有60%的患者疼痛症状得到缓解。盂唇(91%,P0.003)或LCP(89%,P0.05)患者疗效较好,而ON和DJD患者疼痛症状缓解较差,改善率仅为40%和44%。在吻合血管游离腓骨移植(free-vascularized fibular graft,FVEG)的患者中有34%在随访期间得到改善(P=0.003)。其中18名患者(21%)进行了全髋关节置换术。结论:髋关节镜手术对于游离体、盂唇损伤、局灶性软骨病变、晚期LCP后遗症患者有良好的治疗效果;对股骨头坏死的治疗效果不佳。     

尼莫地平在预防缺血性脑血管事件所引起的认知障碍中的作用

目的:观察尼莫地平治疗预防缺血性脑血管事件所引起的认知障碍中的临床效果。方法:将在我院进行治疗的120例缺血性脑血管事件患者随机分为观察组60例和对照组60例。对照组患者按照常规方法进行治疗,观察组患者在此基础上给予尼莫地平治疗,观察对比两组的治疗效果。结果:观察组总有效率显著高于对照组,差异具有统计学意义(P0.05)。两组患者治疗后MOCA评分和MMSE评分均随着治疗时间的延长而逐渐增加。对照组患者治疗2个月及3个月后MOCA评分与治疗前比较,差异具有统计学意义(P0.05);观察组治疗1个月后MOCA评分与治疗前比较,差异具有统计学意义(P0.05)。观察组每月治疗后MOCA评分显著高于对照组,差异具有统计学意义(P0.05)。两组患者治疗2个月及3个月后MMSE评分与治疗前比较,差异具有统计学意义(P0.05)。观察组患者治疗3个月后MMSE评分显著高于对照组,差异具有统计学意义(P0.05)。随访1年后,观察组MMSS、MOCA评分均显著高于对照组,差异均具有统计学意义(P0.05)。两组患者治疗期间均未出现严重不良反应。结论:尼莫地平对于缺血性脑血管事件所引起的认知障碍的预防有较好疗效,安全性良好,值得推广应用。     

NPR3 protects cardiomyocytes from apoptosis through inhibition of cytosolic BRCA1 and TNF-α

Natriuretic peptide receptor 3 (NPR3) is a clearance receptor by binding and internalizing natriuretic peptides (NPs) for ultimate degradation. Patients with cardiac failure show elevated NPs. NPs are linked to poor long-term survival because of their apoptotic effects. However, the underling mechanisms have not been identified yet. Here we report the role of NPR3 in anti-apoptosis via the breast cancer type 1 susceptibility protein (BRCA1) and tumor necrosis factor α (TNF-α ). To demonstrate a role for NPR3 in apoptosis, stable H9C2 cardiomyocyte cell lines using shRNA to knockdown NPR3 were generated. The activities of caspase-3, 8, and 9 were significantly increased in NPR3 knockdown H9C2 cardiomyocytes. Knockdown of NPR3 increased the expression of BRCA1. Also NPR3 knockdown remarkably increased the activity of cAMP response element-binding protein (CREB), a positive regulatory element for BRCA1 expression. BRCA1 showed dispersed nuclear localization in non-cardiomyocytes while predominantly cytoplasmic localization in H9C2 cells. Meanwhile, NPR3 knockdown significantly increased TNF-α gene expression. These data show that NPR3 knockdown in H9C2 cells triggered both extrinsic and intrinsic apoptotic pathways. NPR3 protects cardiomyocytes from apoptosis through inhibition of cytosolic BRCA1 and TNF-α, which are regulators of apoptosis. Our studies demonstrate anti-apoptosis role of NPR3 in protecting cardiomyocytes and establish the first molecular link between NP system and programmed cell death.