TRIM59 通过结合 BCLAF1 调控人皮肤黑色素瘤 SK-MEL-2 细胞的恶 性生物学行为

目的：探究三结构域蛋白59（TRIM59）调控人皮肤黑色素瘤细胞SK-MEL-2增殖、细胞周期、凋亡及迁移侵袭的作用机制，及其与Bcl2相关转录因子1（BCLAF1)之间的关系。方法：qPCR和WB法检测人表皮黑色素细胞HEMn-LP、人皮肤黑色素瘤细胞SK-MEL-2、UACC903、A375及36例邢台市人民医院2019年2月至2021年7月收集的皮肤黑色素瘤组织中TRIM59的mRNA和蛋白表达，使用脂质体将si-con、si-TRIM59转染至SK-MEL-2细胞中，WB法检测干扰TRIM59表达对细胞中周期蛋白D1（CCND1）、细胞周期素依赖性激酶2（CDK2）、肿瘤抑制蛋白基因（TP53）和 BCLAF1 蛋白表达的影响，CCK-8法、流式细胞术、划痕愈合实验、Transwell实验检测对细胞的活性、凋亡、迁移和侵袭的影响，免疫共沉淀（Co-IP）实验检测对细胞中TRIM59蛋白与BCLAF1结合能力的影响。结果：与HEMn-LP细胞相比，SK-MEL-2、UACC903、A375细胞中TRIM59 mRNA和TRIM59、BCLAF1蛋白均呈高表达（均P<0.05），SK-MEL-2细胞中TRIM59表达水平最高。相较于si-con组和Normal组，沉默TRIM59后，SK-MEL-2细胞的活性显著降低，细胞周期阻滞于G2期，CCND1、CDK2的蛋白表达显著降低，TP53蛋白和细胞凋亡率均显著升高，划痕抑制率明显升高，迁移侵袭细胞数明显降低（均P<0.05）。免疫共沉淀实验结果显示，TRIM59与BCLAF1之间存在蛋白结合关系。TRIM59与 BCLAF1 在肿瘤组织中的表达呈显著的正相关（r=0.878，P<0.001）。结论：干扰TRIM59表达能够抑制人皮肤黑色素瘤SK-MEL-2细胞的增殖、迁移和侵袭而促进凋亡，抑制SK-MEL-2细胞的恶性生物学行为，其机制可能与TRIM59结合BCLAF1有关。     

电针刺激对脊髓挫伤模型大鼠神经细胞凋亡的影响

BACKGROUND: Electroacupuncture has promoting effects on the functional recovery of the injured spinal cord, can decrease pain, and elevate postoperative effect after acute spinal cord contusion.  OBJECTIVE: To observe the effect of electroacupuncture on apoptosis in the injured site after spinal cord contusion, and analyze its neuroprotective effects on neurological function in rats with spinal cord contusion. METHODS: A total of 66 adult female Sprague-Dawley rats were randomly divided into: sham surgery group (n=20), spinal cord contusion group (n=20), electroacupuncture stimulation group (n=20) because six rats were excluded due to modeling failure and death. Before model establishment, at 1, 3 days, 1, 2, 3 and 4 weeks after model establishment, motor functions were evaluated by BBB score and the inclined plate test. At 3 days after model establishment, apoptosis of nerve cells could be detected in the site of injury in each experimental group using TUNEL assay. mRNA and protein expression of bax, bcl-2 and caspase-3 was detected surrounding the injury site using RT-PCR and western blot assay. Morphological changes in the site of injury could be observed using hematoxylin-eosin staining. The regeneration of nerve fibers was observed using HRP tracing.  RESULTS AND CONCLUSION: (1) Motor function score was significantly increased at various time points in the 2nd week of treatment in the electroacupuncture stimulation group than in the spinal cord contusion group (P < 0.05). (2) Apoptotic index was significantly lower in the electroacupuncture stimulation group than in the spinal cord contusion group at 3 days after model establishment (P < 0.05). (3) mRNA and protein expression of bax and caspase-3 was significantly lower in the electroacupuncture stimulation group than in the spinal cord contusion group at 72 hours (P < 0.05); bcl-2 gene and protein expression was significantly higher (P < 0.05). (4) The number of HRP-positive nerve fibers was highest in the sham surgery group, followed by electroacupuncture stimulation group, and lowest in the spinal cord contusion group at 4 weeks (P < 0.05). Results indicated that electroacupuncture plays a protective role on the spinal cord contusion by reducing apoptosis of nerve cells at the site of injury.      

异丙酚预处理联合脐血间充质干细胞移植治疗脑缺血再灌注损伤

BACKGROUND:As propofol has a neuroprotective effect, and umbilical cord blood mesenchymal stem cells have a high differentiation potential, their combination will have a better therapeutic effect on cerebral ischemia-reperfusion injury. OBJECTIVE:To study the effect of propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation on cerebral ischemia-reperfusion injury in rats. METHODS:Sixty-three Sprague-Dawley rats were randomized into model, propofol, and combined group (n=21 per group). Rat models of cerebral ischemia-reperfusion injury were made using ligation of the middle cerebral artery occlusion in the three groups. Rats in the combined group were given 100 mg/kg propofol injection at 1 day before injury and injection of umbilical blood mesenchymal stem cells via the tail vein (0.5 mL, 2×10⁹/L). RESULTS AND CONCLUSION:Compared with the model group, the neurological function was improved significantly in the propofol and combined group, especially in the latter one, presenting with a remarkable mitigation in brain injury and an increased level of survivn mRNA in the rat hippocampus. The content of serum malondialdehyde was lower but the activity of superoxide dismutase was higher in the combined group compared with the propofol group. These findings indicate that propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation has better therapeutic effects than propofol pretreamtnet alone for improving cerebral ischemia-reperfusion injury in rats.     

电针刺激对神经干细胞移植治疗脊髓损伤大鼠后肢功能的影响

背景：单纯的神经干细胞移植对受损脊髓组织的修复作用并不理想,为了进一步提高移植细胞在体内的存活、增殖及定向分化为神经元的比例,必须进一步改善脊髓损伤区的微环境。 目的：观察神经干细胞移植联合电针刺激对脊髓损伤大鼠后肢功能及电生理的影响。 方法：将脊髓损伤模型SD大鼠72只按随机数字表法分为4组：对照组尾静脉注入培养液,神经干细胞组经尾静脉注入等体积神经干细胞悬液,电针刺激组自模型完成6 h起采用督脉加体穴电针1周,联合组尾静脉注射神经干细胞后,同时采用督脉加体穴电针1周。分别于造模前、造模后1,3 d、1-4 周通过BBB评分、斜板试验进行运动功能评定。造模后4周取材行病理切片苏木精-伊红染色,荧光显微镜观测CM-Dil 标记的神经干细胞存活及分布情况,辣根过氧化物酶示踪观察神经纤维再生情况,运动诱发电位和体感诱发电位观察大鼠神经电生理恢复情况。 结果与结论：造模后2-4周大鼠下肢运动功能评价联合组优于神经干细胞组及电针刺激组,神经干细胞组和电针刺激组优于对照组。造模后4周,神经干细胞组和电针刺激组损伤区可见少量神经轴索样结构,脊髓空洞较小,联合组可见较多神经轴索样结构,未见脊髓空洞。造模后4周,CM-Dil 阳性细胞和辣根过氧化物酶阳性神经纤维数：联合组>神经干细胞组与电针刺激组>对照组,各组之间差异有显著性意义(P < 0.05)。运动诱发电位和体感诱发电位的潜伏期：联合组<神经干细胞组与电针刺激组<对照组,各组之间差异有显著性意义(P < 0.05);运动诱发电位和体感诱发电位的波幅：联合组>神经干细胞组与电针刺激组>对照组,各组之间差异有显著性意义(P < 0.05)。结果提示神经干细胞移植的同时联合电针刺激能够促进脊髓损伤大鼠神经突触的再生,改善其大鼠肢体运动功能及电生理功能。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程      

丙泊酚对脐带间充质干细胞移植治疗大鼠脊髓损伤后肢功能变化的影响

目的探讨脊髓损伤模型大鼠单独移植人脐带间充质干细胞(UC-MSCs)、单独应用丙泊酚治疗及联合使用UCMSCs和丙泊酚之后,对其电生理的改变及脊髓损伤后肢功能的影响。方法纳入70只SD大鼠,在进行正式实验前将所有大鼠提前制成脊髓损伤模型,然后随机将存活下来的已制备成模型的SD大鼠分为对照组(n=16)、UC-MSCs组(n=16)、丙泊酚组(n=16)、联合组(n=16)。斜板试验、Basso Beattie Bresnahan(BBB)评分被应用于实验前1 d及实验后1、3 d、1、2、3、4 w,共6次,用来评价脊髓损伤模型大鼠的下肢运动功能水平。实验第4周,CM-Dil被应用于观察获取样本的UC-MSCs存活率,并且应用HE染色观察病理切片UC-MSCs分布情况。此外,随机抽取每组大鼠,分别对其进行辣根过氧化物酶(HRP)束路示踪法和体感诱发电位(SEP)和运动诱发电位(MEP)的检测,观察大鼠神经纤维再生及神经电生理的恢复情况。结果通过对SD大鼠后肢运动功能进行评定并且比较后,联合组的功能优于其他组,但对照组是4组中功能最弱的。HRP束路示踪法所观察到的神经纤维再生恢复情况与CM-Dil标志的UC-MSCs存活率相似,对照组相对其他三组,数值最低,而UC-MSCs组和丙泊酚组也略低于联合组。通过HE染色的应用,可对病理切片观察到,对照组脊髓损伤明显,无神经轴索通过,并且可见脊髓空洞的形成,组织也部分缺失,而经过单纯UC-MSCs移植和单纯应用丙泊酚的大鼠,对比对照组,尽管也有部分损伤,但可见少许神经轴索,联合组的切片比其他三组均病理化程度较低,神经轴索较多且不可见脊髓空洞。MEP、SEP均显示联合组优于UC-MSCs组及丙泊酚组,但经过治疗的三组又优于对照组。结论丙泊酚对大鼠脊髓损伤有着神经保护的作用,UC-MSCs移植的同时泵注丙泊酚可以促进大鼠脊髓损伤神经的再生突触,对下肢的神经生理功能有着极大的改善。     

颈肩部手术患者改良肌间沟法颈丛阻滞的效果

颈丛阻滞是颈肩部手术常用的麻醉方法,传统的颈深丛或浅丛阻滞多采用C4横突注射法,而此法局麻药扩散较局限,阻滞效果不甚理想,且不良反应发生机率较高.本研究拟评价改良肌间沟法颈丛阻滞的效果,为临床提供参考.     

不同体位下股骨近端防旋髓内钉内固定治疗股骨转子间骨折肥胖患者效果观察

目的观察肥胖患者发生股骨转子间骨折时取平卧位与侧卧位下股骨近端防旋髓内钉（PFNA）内固定治疗的效果。方法 2010年1月—2013年6月河北医科大学附属邢台市人民医院采用PFNA治疗股骨转子间骨折肥胖患者（体质量指数〉26 kg/m^2）43例,根据体位不同分为平卧位组（21例）与侧卧位组（22例）。比较两组术中出血量、手术时间、转子顶端切口长度、住院时间、开始负重时间、骨折愈合时间、髋关节Harris评分及术后并发症发生率。结果侧卧位组手术时间、转子顶端切口长度较平卧位组短,术中出血量较平卧位组少,差异有统计学意义（P〈0.05）。两组住院时间、开始负重时间、骨折愈合时间、髋关节Harris评分比较差异无统计学意义（P〉0.05）。侧卧位组并发症发生率（13.6%）明显低于仰卧位组（38.1%）,差异有统计学意义（P〈0.05）。结论对于股骨转子间骨折肥胖患者,侧卧位下行PFNA内固定术能充分暴露术区、有效降低手术难度、减少术中出血量、缩短手术切口长度和手术时间,较平卧位有明显优势。