Long-term hypothermia has been reported to prevent intracranial pressure (ICP) rebound in clinical patients, but the duration for hypothermia and the corresponding ICP data are not available. This study investigated the optimal duration of long-term hypothermia in traumatic brain injury (TBI) rats, and observed the effect on ICP and neurological function. In this study, we established a rat severe TBI model with electronic Controlled Cortical Injury device, and implemented hypothermia (33 °C) for different durations. The motor function of the rats in each group was evaluated by beam walking test and inclined-grid climbing test, brain water content was calculated by the wet-dry weight method, Evan's blue staining was used to measure the blood-brain barrier (BBB) permeability, the change of hippocampal neurons was observed by Nissl staining, the expressions of BrdU, NeuN, and CD86 positive cells were detected by immunofluorescence staining, and the expressions of Bcl-2, Bax, iNOS, IL-10, and Arg-1 were detected by Western blot. We found that therapeutic hypothermia improved neurological recovery after TBI with declining ICP, reducing brain edema, decreasing BBB permeability, promoting neurogenesis, inhibiting apoptosis, and regulating inflammation. Moreover, 48 h hypothermia amplified the neuroprotective effect after injury on the basis of 4 or 24 h hypothermic treatment. Both 4 and 24 h hypothermia led to ICP rebound during or after rewarming, whereas 48 h hypothermia completely abolished ICP rebound. Our study suggests that long-term hypothermia amplifies neuroprotection after TBI by antagonizing ICP rebound.
目的 探究经颅直流电刺激(tDCS)联合右佐匹克隆治疗慢性失眠的临床应用价值.方法 68例慢性失眠患者分为对照组35例(接受tDCS假刺激联合右佐匹克隆治疗)和研究组33例(接受tDCS联合右佐匹克隆治疗),连续治疗4周.比较2组治疗后临床疗效、多导睡眠监测(PSG)结果、血清皮质醇(CORT)、促肾上腺皮质激素(ACTH)水平及匹兹堡睡眠质量指数量表(PSQI)、抑郁症9项筛查量表(PHQ-9)、广泛性焦虑量表-7(GAD-7)评分及不良反应.结果 治疗4周,研究组治疗总有效率93.94%(31/33)高于对照组的74.29%(26/35)(P＜0.05).治疗2周、4周,与对照组比较,研究组总睡眠时间(TST)延长,睡眠潜伏期(SL)变短,N3期睡眠比例、睡眠效率(SE)升高,微觉醒指数、N1期睡眠比例、血清CORT、ACTH水平均降低(P＜0.05).治疗4周和治疗后6个月,与对照组比较,研究组PSQI、PHQ-9评分均降低(P＜0.05).治疗2周、4周及治疗后6个月,与对照组比较,研究组GAD-7评分降低(P＜0.05).2组不良反应发生率比较差异无统计学意义(P＞0.05).结论 tDCS联合右佐匹克隆治疗可提高慢性失眠患者睡眠质量,降低焦虑抑郁程度,并具有远期疗效.其机制可能与下调患者血清CORT、ACTH水平有关.
目的 探讨MDA-MB-231乳腺癌细胞培养上清液对人脂肪间充质干细胞(hAdMSC)迁移、增殖和凋亡的影响及分子机制.方法 将MDA-MB-231细胞上清液和不含胎牛血清的RPMI-1640培养基以1∶4的体积比混匀后培养的hAdMSC为MDA-MB-231上清液组.向MDA-MB-231上清液组添加10µmol/L Reparixin(CXCR1/2抑制剂)为CXCR1/2抑制剂组;向MDA-MB-231上清液组添加10 nmol/L GSK690693(Akt抑制剂)为Akt抑制剂组.无任何刺激进行培养的hAdMSC为对照组.细胞划痕实验检测各组hAdMSC的迁移能力,CCK-8实验检测各组hAdMSC增殖情况,Annexin V-FITC/PI双标记流式细胞凋亡实验检测各组hAdMSC凋亡率,Western blot检测各组hAdMSC的mTOR/磷酸化mTOR(p-mTOR)和Akt/磷酸化Akt(p-Akt)蛋白表达.结果 与对照组相比,MDA-MB-231上清液组hAdMSC的24h和48h细胞划痕闭合面积、细胞增殖水平以及p-Akt和p-mTOR蛋白相对表达量均增加,细胞凋亡率降低(P＜0.05);与MDA-MB-231上清液组相比,Akt抑制剂组和CXCR1/2抑制剂组hAdMSC的48h细胞划痕闭合面积、细胞增殖水平以及p-Akt和p-mTOR蛋白相对表达量均降低,细胞凋亡率均增加(P＜0.05).结论 MDA-MB-231乳腺癌细胞培养上清液通过激活Akt-mTOR信号通路促进hAdMSC迁移和增殖,抑制hAdMSC凋亡,其中IL-8-CXCR1/2轴发挥关键作用.
目的 探讨肿瘤相关巨噬细胞(TAM)对三阴性乳腺癌(TNBC)细胞白蛋白紫杉醇(Nab-PTX)化疗敏感性的影响及作用机制.方法 构建并鉴定TAM模型;通过Transwell小室共培养法建立TAM与TNBC细胞系MDA-MB-231细胞共培养模式,分为对照组(MDA-MB-231细胞及空白小室)、Nab-PTX组(MDA-MB-231细胞、空白小室及0.5 nmol/L Nab-PTX)、TAM组(MDA-MB-231细胞、含M2型巨噬细胞的小室)、TAM+Nab-PTX组(MDA-MB-231细胞、含M2型巨噬细胞的小室及0.5 nmol/L Nab-PTX)、胰岛素样生长因子1受体(IGF-1R)抑制剂组(MDA-MB-231细胞、含M2型巨噬细胞的小室、0.5 nmol/L Nab-PTX及4 nmol/L IGF-1R抑制剂Linsitinib);CCK-8法检测各组细胞增殖情况;流式细胞术检测细胞凋亡;实时荧光定量PCR(qPCR)检测多药耐药蛋白(MDR)1和胱天蛋白酶(Caspase)-3 mRNA水平;Western blot检测IGF-1R信号通路关键蛋白表达.结果 THP-1细胞经诱导分化为M2型巨噬细胞;与Nab-PTX组相比,TAM+Nab-PTX组细胞增殖水平升高,凋亡率降低(P＜0.01),MDR1 mRNA表达升高,Caspase-3 mRNA表达降低(P＜0.05),IGF-1R信号通路关键蛋白激活(P＜0.01);与TAM+Nab-PTX组相比,IGF-1R抑制剂组细胞增殖水平降低,凋亡率升高,MDR1 mRNA表达降低,Caspase-3 mRNA的表达增高,IGF-1R信号通路关键蛋白的表达降低(P＜0.01).结论 TAM可能通过激活IGF-1R信号通路诱导TNBC细胞对Nab-PTX耐药.
目的 探讨CircBACH1对多发性骨髓瘤(MM)细胞增殖、凋亡和化疗敏感性的影响以及在此过程中对miR-140-5p/MDM2轴的调节机制.方法 收集MM组和正常对照组骨髓浆细胞,将人MM细胞系U266细胞分为未转染(Control)组、si-NC组、si1-CircBACH1组、si2-CircBACH1组、si-CircBACH1+anti-miR-NC组、si-CircBACH1+ anti-miR-140-5p组.实时荧光定量PCR检测细胞中CircBACH1、miR-140-5p、鼠双微体2(MDM2)mRNA的表达;CCK-8法检测细胞增殖活性;流式细胞术检测细胞凋亡及对硼替佐米的敏感性;Western blot检测细胞中B细胞淋巴因子2(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解的胱天蛋白酶(cleaved caspase)-3蛋白的表达;双萤光素酶报告基因实验验证CircBACH1、miR-140-5p、MDM2三者的靶向关系.结果 与正常对照组相比,MM组的骨髓浆细胞及U266细胞中CircBACH1、MDM2 mRNA表达升高,miR-140-5p表达降低(P＜0.05);与Control组和si-NC组相比,si-CircBACH1组细胞中CircBACH1、MDM2的mRNA表达、细胞增殖活性、Bcl-2蛋白表达均降低,miR-140-5p的表达、细胞凋亡率、化疗敏感性及cleaved caspase-3、Bax表达均升高(P＜0.05);而回补实验中,si-CircBACH1对MM细胞的影响被anti-miR-140-5p逆转,且MDM2的表达也升高(P＜0.05).双萤光素酶基因报告实验验证了CircBACH1、MDM2均与miR-140-5p存在靶向关系.结论 敲低CircBACH1能够上调miR-140-5p表达,下调MDM2的表达,抑制MM细胞的增殖,促进MM细胞凋亡,提高化疗敏感性.
