The role of autophagy in the transplantation of induced pluripotent stem cells (iPSCs)-derived neural stem cells (NSCs) to treat spinal cord injury (SCI) and neurogenic bladder was investigated in this study. NSCs derived from human iPSCs were identified by and immunofluorescence assay. To clarify the role of autophagy, iPSCs were treated with either an autophagy inducer (rapamycin), or an autophagy inhibitor (chloroquine). Cell Counting kit-8 (CCK-8), western blot and flow cytometry were used to detect the effect of autophagy on the viability and differentiation of iPSCs. Sixty Wistar rats were selected to establish the SCI model and treated with iPSCs-derived NSCs transplantation. The effect of autophagy on the bladder function of rats with different treatments was evaluated by Basso, Beattie, and Bresnahan (BBB) score, bladder function score, bladder weight measurement, Hematoxylin & Eosin (H&E) staining, and Masson staining. The results of in vitro experiment showed that rapamycin enhanced the cell activity of iPSCs, increased the number of nestin positive cells, up-regulated Beclin-1 and LC3BI/II expressions, and down-regulated p62 expression. And the results of in vivo experiment showed that rapamycin improved exercise ability and bladder function, partially restored bladder weight, and significantly reduced bladder tissue damage in SCI rats. However, chloroquine showed the opposite results. The differentiation of iPSCs into NSCs could be promoted by induced autophagy, while neurogenic bladder of SCI was restored by autophagy activation.

目的 探讨关节镜下改进双Endobutton钢板治疗RockwoodⅢ～Ⅴ肩锁关节脱位的疗效.方法 回顾性分析2016年8月至2019年1月24例RockwoodⅢ～Ⅴ型肩锁关节脱位患者临床资料,所有患者采用关节镜下改进双Endobutton钢板行喙锁韧带重建术.观察关节功能恢复情况,采用Bjerneld肩锁关节脱位评价系统进行术后肩锁关节功能评价.结果 24例患者术后获得随访,时间8～24个月,平均(11.6±2.4)个月.所有患者肩部外形均正常,无肩锁关节复位丢失.锁骨及喙突无医源性骨折.根据Bjerneld肩锁关节脱位评价系统:优20例,良4例.结论 RockwoodⅢ～Ⅴ型肩锁关节脱位患者,关节镜下改进双Endobutton钢板内固定重建喙锁韧带,具有微创,医源性骨折风险小,局部稳定性佳,肩锁关节功能恢复满意.

目的 探讨脊柱内窥镜下腰脊神经背内侧支射频消融术治疗中老年慢性腰椎小关节源性腰痛的近期疗效.方法 采用回顾性队列研究的方法,选取2015年9月—2017年9月杭州市中医院收治的28例慢性腰椎小关节源性腰痛患者,其中14例采用脊柱内窥镜下腰脊神经背内侧支射频消融术治疗(手术组),14例采用经皮腰椎关节突关节封闭治疗(非手术组).分别对2组患者进行电话、门诊随访,记录治疗前及治疗后1周、1个月、3个月、6个月、1年的腰痛视觉模拟量表(VAS)评分、日本骨科学会(JOA)评分和Oswestry功能障碍指数(ODI)并进行比较,评估2种治疗方案近期疗效的差异.结果 治疗前2组患者VAS评分、JOA评分和ODI差异均无统计学意义(P>0.05).治疗后2组患者各时间点VAS评分、JOA评分和ODI均较治疗前改善,其中手术组与治疗前相比差异有统计学意义(P<0.05);手术组患者VAS评分、JOA评分和ODI的改善情况均优于非手术组,差异有统计学意义(P<0.05).结论 脊柱内窥镜下腰脊神经背内侧支射频消融术治疗慢性腰椎小关节源性腰痛能够显著缓解患者症状、改善预后,近期疗效较好,相比经皮腰椎关节突关节封闭具有一定优势.

腰骶分叉神经是一种解剖变异,随着影像技术及脊柱内镜技术的发展,腰骶分叉神经逐渐引起临床重视.脊柱内镜手术的视野范围相对较小,若术前未发现存在腰骶分叉神经,术中容易造成神经损伤,因此提高腰骶分叉神经的诊断率是手术成功的重要保障.本文从脊柱内镜手术神经损伤的发生率及腰骶分叉神经的解剖结构、分型、发生率、影像诊断几个方面对腰骶分叉神经的研究进展进行了综述.

Objective To investigate the effect of hypoxia on acid sensitive ion channel 3 (ASIC3) in nucleus pulposus microenvironment and its mechanism.Methods Primary nucleus pulpocytes were isolated and identified by immunohistochemistry,hematoxylin and eosin (HE) and toluidine blue.Immunohistochemistry,HE,toluidine blue and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were used to explore the appropriate hypoxic concentration,apoptosis and FQ-PCR to explore the appropriate action time.The interference and overexpression of ASIC3 vector were constructed,and their efficiency was verified by FQ-PCR and Western blotting.The experiment was divided into the normal group (Control).Hypoxic treatment group (2％ O2);shRNA-NC + 2％ O2);shRNA-ASIC3 + 2％ O2;Expression of no-load + hypoxic treatment (Vector + 2％ O2);ASIC3 expression + hypoxic treatment group (ASIC3 + 2％ O2).The proliferation was detected by cell counting kit-8 (CCK-8),the cell cycle and apoptosis were detected by flow cytometry,the formation of autophagosomes in each group was detected by transmission electron microscopy,the expression of hypoxia inducible factor (H1F)-1 and ASIC3 was detected by immunofluorescence,the expression of HIF-1,light chain 3 (LC3),extracellular signal-regulated kinase 1/2 (ERK1/2) and mitogen-activated protein kinase (MAPK) and phosphorylation were detected by FQ-PCR and Western blotting,and the effect of ASIC3 on HIF-1 and ERK1/2 was detected by immunoco-precipitation.Results Nucleus pulposus cells were successfully isolated with appropriate hypoxic concentration of 2％ and appropriate action time of 48 h.Both interference and overexpression vector construction were successful and effective.Compared with the control group,cell proliferation was inhibited in the hypoxic group and ASIC3 expression + hypoxic treatment group,apoptosis was increased,cell cycle arrest was in G1 phase,calcium ion concentration was increased,and HIF-1,LC3,p -ERK1/2 and p-mapk expression were increased,the difference was statistically significant (P ＜ 0.05).Conclusion Interference with ASIC3 can inhibit the activation of mitogen-activated protein kinase (MAPK) signal to improve the microenvironment of hypoxic nucleus pulposus cells,which may be one of the pathogenesis of discogenic low back pain.

通常,MRI发现的腰椎间盘突出的位置与下肢放射痛的症状是相对应的.但是,有学者报道临床上有部分腰椎间盘突出症患者表现为下肢症状与椎间盘突出侧左右不一致的特殊现象.这种现象产生的原因尚没有定论,给诊断和治疗带来困难.笔者根据国内外类似病例的文献报道和临床经验,探讨了该现象形成的原因、病理机制以及不同的治疗方案并作一综述.

腰椎关节突关节病变引起的腰痛称为腰椎关节突关节源性腰痛,目前关于该病的报道较少,在发病率及最佳诊断方法、治疗方法方面还存在争议.本文从发病机制、诊断、治疗3个方面对腰椎关节突关节源性腰痛的研究进展进行了综述,以期为临床诊治该病提供参考.

Objective To investigate the effect of amiloride on the expression of acid sensing ion channel 3 (ASIC3) in dorsal root ganglion (DRG) following nerve root inflammation induced by nucleus pulposus in rats.Methods Forty eight SD rats were randomly divided into control group(Sham,n=16),inflammation group (NP,n=16),inflammation+amiloride group (NP+Ami,n=16).The L5 DRG was exposed in rats of NP and NP+Ami groups by surgery,and sham surgery was performed in sham group.Intraperitoneal injection of amiloride 20μg/kg was given to rats in NP+Ami group q.d for 2 weeks after surgery,and no amiloride was given to rats in sham group and NP group.The mechanical pain withdrawal threshold (PWT) was evaluated at 2,4,6,8,10 and 12d after modeling.The expression of ASIC3,TNF-α,IL-6 mRNA and protein were examined by Real-time PCR and Western blot,respectively.TNF-α and IL-6 concentrations present in peripheral blood and DRG were determined at 1,3,7 and 14d after the surgical procedures using ELISA method.NO levels were examined by biochemistry test.Results Rats exposed to the NP showed decreased PWT at 12d after surgery (P＜0.01),and the levels of ASIC3 and the NO,TNF-α,IL-6 were up-regulated in DRG induced by NP 14d after surgery(all P＜0.01).Compared to NP group the mechanical PWT decreased at 8d after surgery and then gradually increased at 10 and 12d after surgery in NP+Ami group (all P＜0.01).ASIC3 was down-regulated (both P＜0.05),and the levels of NO,TNF-α,IL-6 were significantly decreased at 14d after surgery (all P＜0.01) in NP+Ami group.Conclusion ASIC3 may upgrade the expression of NO and inflammatory cytokines TNF-α and IL-6 to induce hyperalgesia and never root pain.