川芎嗪对大鼠神经病理性痛的影响

目的 评价川芎嗪对大鼠神经病理性痛的影响.方法 成年健康雄性SD大鼠54只,体重180～ 220 g,采用随机数字表法,将其随机分为3组(n=18):假手术组(S组)、神经病理性痛组(NP组)和川芎嗪组(T组).S组仅分离坐骨神经但不结扎,NP组和T组采用坐骨神经慢性压迫损伤法建立神经病理性痛模型.T组于术毕开始腹腔注射川芎嗪100 mg/kg,1次/d,连续14 d;S组和NP组以等容量生理盐水替代.于术前1d、术后3、7、14 d时测定机械痛阈和热痛阈,并于术后3、7和14d时测定痛阈后各取6只大鼠处死,取L(4).5脊髓组织,用免疫组织化学法检测脊髓背角Bcl-2及caspase-3的表达,TUNEL法检测脊髓背角凋亡细胞,计算细胞凋亡率.结果 S组比较,NP组和T组热痛阈及机械痛阈降低,脊髓背角Bcl-2和caspase-3表达上调,细胞凋亡率升高(P＜0.05).与NP组比较,T组热痛阈及机械痛阈升高,脊髓背角Bcl-2表达上调,caspase-3表达下调,细胞凋亡率降低(P＜0.05).结论 川芎嗪可减轻大鼠神经病理性痛,其机制与抑制脊髓背角细胞凋亡有关.     

脊髓背角膜结合型补体调节蛋白表达在大鼠神经病理性痛形成中的作用

目的 评价脊髓背角膜结合型补体调节蛋白表达在大鼠神经病理性痛形成中的作用.方法 健康雄性SD大鼠,体重200 ～ 250 g.取48只鞘内置管成功的大鼠,采用随机数字表法,将其分为4组(n=12):假手术组(S组)、神经病理性痛组(NP组)、生理盐水组(NS组)和米诺环素组(M组).NP组、NS组和M组采用坐骨神经环形结扎法制备大鼠神经病理性痛模型,S组只分离坐骨神经后逐层缝合.M组于结扎坐骨神经前ld开始鞘内注射米诺环素50 μg,NS组鞘内注射等容量生理盐水,1次/d,连续7d.于结扎坐骨神经前1 d(T0)、结扎后1 d(T1)、3 d(T2)和7 d(T3)时测定大鼠机械痛阈和热痛阈.于T3痛阈测定结束后断头处死大鼠,取L4.5脊髓背角组织,分别用Western blot法和RT-PCR法检测CD46、CD55、CD59蛋白及其mRNA的表达水平.结果 与S组比较,NP组、NS组和M组T1-3时机械痛阈和热痛阈降低,T3时脊髓背角CD46、CD55、CD59蛋白及其mRNA表达下调(P＜0.05);与NS组和NP组比较,M组T2.3时机械痛阈和热痛阈升高,T3时脊髓背角CD46、CD55、CD59蛋白及其mRNA表达上调(P＜0.05).结论 脊髓背角膜结合补体调节蛋白表达下调,补体异常活化参与了大鼠神经病理性痛的形成.     

鞘内注射DREAM-shRNA对神经病理性痛大鼠脊髓背角p-CREB表达的影响

目的 探讨鞘内注射转录因子下游调控元件拮抗因子-短发夹RNA(DREAM-shRNA)对神经病理性痛大鼠脊髓背角磷酸化环磷酸腺苷反应元件结合蛋白(p-CREB)表达的影响.方法 成年健康雄性SD大鼠,体重280～320 g,采用坐骨神经慢性压迫(CCI)法建立大鼠神经病理性痛模型.于CCI后第3天鞘内置管.取鞘内置管成功的大鼠24只,随机分为4组,每组6只,假手术组(S组):仅暴露坐骨神经,不结扎;神经病理性痛组(NP组):于CCI后第8天鞘内注射生理盐水10 μl;RNA干扰组(RNAi组):于CCI后第8天鞘内注射DREAM-shRNA 5 μl和牛理盐水5 μl;空白载体组(BV组):于CCI后第8天鞘内注射慢病毒空白载体5 μl和生理盐水5μl,各组连续注射7 d.于CCI前1 d(T0,基础状态)、CCI后第7～14天(T1-8)时测定机械痛阈.于CCI后第15天时测定脊髓背角绿色荧光蛋白(GFP)和p-CREB的表达水平.结果 与基础值比较,各组各时点机械痛阈降低(P<0.05或0.01);与T1时比较,NP组和BV组T5-8时机械痛阈降低,RNAi组T8时机械痛阈升高(P<0.05或0.01);与S组比较,NP组和BV组机械痛阈降低,RNAi组T2时机械痛阈降低,T8时升高,NP组、RNAi组和BV组脊髓背角p-CREB表达上调(P<0.05);与NP组比较,RNAi组T6-8时机械痛阈升高,脊髓背角p-CREB表达下调(P<0.05).RNAi组脊髓背角可见大量绿色荧光,即GFP表达阳性,其余3组脊髓背角未见绿色荧光,即GFP无表达.结论 鞘内注射DREAM-shRNA缓解大鼠神经病理性痛的机制可能与抑制脊髓背角p-CREB的表达有关.     

鞘内注射PSD-93反义寡核苷酸对大鼠神经病理性痛的疗效

目的 探讨鞘内注射PSD-93反义寡核苷酸对大鼠神经病理性痛的疗效.方法 雄性SD大鼠60只,随机分为5组(n=12):假手术组(S组)、C7脊神经压迫组(N组)、C7脊神经压迫+鞘内注射PSD-93误义寡核苷酸10 μg组(M组)、C7脊神经压迫+鞘内注射PSD-93反义寡核苷酸5 μg组(AJ组)和C7脊神经压迫+鞘内注射PSD-93反义寡核苷酸10 μg组(A2 组).N组、M组、A1组和A2组用60 g无创微血管夹压迫大鼠右侧C7脊神经15 min,制备神经病理性痛模型,经枕骨大孔鞘内置管,术毕当日开始给药,每日1次,连续4 d.于术前2 d(T0)和术后1、3、5、7 d(T1-4)测定机械痛阈和、热痛阈;T2和T4时分别处死6只大鼠,取C7脊髓,免疫组化法测定脊髓背角PSD-93蛋白表达.结果 与S组比较,N组、M组和A1组T1-4时机械痛阈及热痛阈降低,N组和M组T2,4时脊髓背角PSD-93蛋白表达上调(P＜0.05);与N组和M组比较,A1组T1,2时、A2组T1～4时机械痛阈及热痛阈升高,A1组T2时、A2组T2,4时脊髓背角PSD-93蛋白表达下调(P＜0.05);与A1组比较,A2组T1-4时机械痛阈及热痛阈升高,T2,4时脊髓背角PSD-93蛋白表达下调(P＜0.05).结论 鞘内注射PSD-93反义寡核苷酸可减轻大鼠神经病理性痛.     

丹参酮ⅡA对糖尿病大鼠神经病理性疼痛的影响

目的研究丹参酮IIA对糖尿病大鼠神经病理性疼痛的影响及其机制。方法18只成年雄性SD大鼠,随机分为3组：正常对照组(C组)、糖尿病组(B组)、糖尿病+IIA组(A组)(n=6)。以佐脲霉菌素(STZ)75mg·kg~(-1)一次性经腹腔注射建立糖尿病模型,分别于注射STZ前、注射后7、14、27d测定机械性痛阈。于A组机械性痛阈小于8g开始腹腔注射丹参酮IIA 25 mg·kg~(-1),每日1次,持续2周。于注射STZ后27d处死大鼠,取I_(?)脊髓,采用尼氏体染色法光镜下观察脊髓组织的形态学变化,应用免疫组化法测脊髓神经元降钙素基因相关肽(CGRP)表达。结果与C组比较,A组、B组机械性痛阈降低(P＜0．05)；与B组比较．A组在注射STZ后27 d机械性痛阈升高(P＜0．05)。B组脊髓背角萎缩．冲经元变性坏死,数量减少．背角神经元内尼氏体减少,溶解消失；与B组比较,A组脊髓背角萎缩减轻,神经元数量增多．可见少量尼氏体。C组脊髓背角神经元有较多CGRP表达；B组背角神经元有少量或者无CGRP表达；与B组比较,A组脊髓背角神经元有大量CGRP表达。结论丹参酮IIA减轻糖尿病大鼠神经病理性疼痛,对脊髓背角神经元有保护作用,其机制可能与丹参酮IIA上调脊髓背角神经元CGRP的表达有关。     

脊髓背角星形胶质细胞NF-κB在紫杉醇诱发大鼠神经病理性痛中的作用

目的 评价脊髓背角星形胶质细胞NF-κB紫杉醇诱发大鼠神经病理性痛中的作用.方法 健康雄性SD大鼠20只,6周龄,体重180～200 g,采用随机数字表法,将其随机分为2组(n＝10):对照组(C组)和神经病理性痛组(NP组).NP组隔日腹腔注射紫杉醇2 mg/kg,共4次;对照组隔日腹腔注射生理盐水1 ml/kg,共4次.于给药前和给药结束后1、7、14 d时分别测定体重、机械痛阈和热痛阈.给药结束后14 d时,处死大鼠,取L4～6脊髓组织,采用免疫荧光法测定脊髓背角星形胶质细胞NF-κB p65的表达.结果 两组体重比较差异无统计学意义(P＞0.05).与C组比较,NP组给药结束后7和14 d时机械痛阈和热痛阈降低,脊髓背角星形胶质细胞NF-κB p65表达上调(P＜0.05或0.01).结论 紫杉醇通过激活脊髓星形胶质细胞中的NF-κB,诱发大鼠神经病理性痛.     

鞘内注射DREAM-shRNA对神经病理性痛大鼠的镇痛效应

目的 评价鞘内注射转录因子下游调控元件拮抗因子-短发夹RNA(DREAM-shRNA)对神经病理性痛大鼠的镇痛效应.方法 健康雄性SD大鼠,体重280-320g,采用坐骨神经慢性缩窄性损伤(CCI)法建立大鼠神经病理性痛模型,于CCI后第3天鞘内置管.取鞘内置管成功的大鼠24只,随机分为4组,每组6只,假手术组(Sham组):仅暴露坐骨神经,不结扎;神经病理性痛组(NP组):于CCI后第8天鞘内注射生理盐水10μl;RNA干扰组(RNAi组):于CCI后第8天鞘内注射含DREAM-shRNA的慢病毒5μl、生理盐水5μl;空白载体组(BV组):于CCI后第8天鞘内注射慢病毒空白载体5μl、生理盐水5μl,连续注射7d.于CCI前1d(基础状态)、CCI后第7～14天(T1-8)测定热痛阈和机械痛阈,于CCI后第15天测定脊髓背角绿色荧光蛋白(GFP)的表达水平.结果 与基础值比较,NP组和BV组热痛阈降低,Sham组T1-4时热痛阈降低,RNAi组T1-4,6时热痛阈降低,4组CCI后各时点机械痛阈降低(P<0.05或0.01);与T1时比较,NP组和BV组其余时点热痛阈和机械痛阈降低.RNAi组T3-5,时热痛阈降低,T1时热痛阈和机械痛阈升高(P<0.05或0.01);与Sham组比较.NP组和BV组热痛阈和机械痛阈降低,RNAi组T2时机械痛阈降低,T5时升高(P<0.05),热痛阈差异无统计学意义(P>0.05);与NP组比较.RNAi组热痛阈和机械痛阈升高(P<0.05).仅RNAi组脊髓背角有GFP表达,其余3组脊髓背角未见GFP表达.结论 鞘内连续注射DREAM-shRNA可在一定程度上缓解大鼠神经病理性痛.     

鞘内注射PSD-93反义寡核苷酸对神经病理性痛大鼠脊髓nNOS的影响

目的 探讨鞘内注射PSD-93反义寡核苷酸对神经病理性痛大鼠脊髓神经型一氧化氮合酶(nNOS)的影响.方法 雄性SD大鼠32只,体重250～350 g,随机分为4组(n=8):假手术+生理盐水组(C组);C7脊神经压迫+生理盐水组(N组);C7脊神经压迫+PSD-93误义寡核苷酸10μg组(M组);C7脊神经压迫+PSD-93反义寡核苷酸10μg组(A组).N组、M组和A组采用60 g微血管夹压迫大鼠右侧C7脊神经15 min制备神经病理性痛模型,经枕骨大孔鞘内置管至颈膨大处.术毕当日开始给药,每日1次,连续4 d.于术前2 d(T0)、术后1、3、5和7 d(T1-4)时测定机械痛阈和热痛阈,术后7 d处死大鼠取C7段脊髓,免疫组化法检测神经压迫侧脊髓PSD-93蛋白和nNOS的表达.结果 与T0时比较,N组和M组各时点机械痛阈及热痛阈降低(P<0.05);与C组比较,N组和M组各时点机械痛阈及热痛阈降低,N组、M组和A组脊髓背角PSD-93蛋白和nNOS表达上调(P<0.05);与N组和M组比较,A组各时点机械痛阈及热痛阈升高,脊髓背角PSD-93蛋白和nNOS表达下调(P<0.05).结论 鞘内注射PSD-93反义寡核苷酸可抑制神经病理性痛大鼠脊髓nNOS表达,nNOS在神经病理性痛中的作用可能受PSD-93蛋白调节.     

氯胺酮与可乐定对神经病理性痛大鼠脊髓P2X4R mRNA表达的影响

目的 评价氯胺酮与可乐定对神经病理性痛大鼠脊髓P2X4受体mRNA(P2X4R mRNA)表达的影响.方法 雄性SD大鼠80只,体重180～220 g,随机分为假手术组(S组)、神经病理性痛组(NP组)、氯胺酮组(K组)、可乐定组(CL组)和氯胺酮+可乐定组(KC组),每组16只.采用坐骨神经慢性压迫(CCI)法制备大鼠神经病理性痛模型.K组、CL组、KC组于CCI后3～21 d每天分别腹腔注射氯胺酮10 mg/kg、可乐定1 mg/kg和氯胺酮5 mg/kg+可乐定0.5 mg/kg.S组和NP组腹腔注射等容量生理盐水.分别于CCI前1 d、CCI后3、7、14和21 d且腹腔注射前,随机取4只大鼠,测定机械痛阈和热痛阈,并于CCI前1 d、CCI后7、14和21 d痛阈测定结束后断头处死,测定脊髓P2X4R mRNA表达水平.结果 与CCI前1 d比较,CCI后S组热痛阈、机械痛阈和脊髓P2X4R mRNA表达差异无统计学意义(P>0.05),其余4组热痛阈和机械痛阈降低,脊髓P2X4R mRNA表达上调(P<0.05);与NP组比较,K组、CL组、KC组CCI后热痛阈及机械痛阈升高,脊髓P2X4R mRNA表达下调(P<0.05);与K组比较,KC组CCI后热痛阈和机械痛阈升高,脊髓P2X4R mRNA表达下调(P<0.05),CL组上述指标差异无统计学意义(P>0.05).结论 氯胺酮与可乐定减轻大鼠神经病理性痛的机制可能与下调脊髓P2X4R mRNA的表达有关.     

曲马多对神经病理性痛大鼠中脑远位触液神经元5-HT1A受体表达的影响

目的 探讨曲马多对神经病理性痛大鼠中脑远位触液神经元5-HT1A受体表达的影响.方法 SPF级雄性SD大鼠40只,体重220～280 g,采用坐骨神经慢性压迫法制备大鼠神经病理性痛模型,随机分为5组(n=8):正常对照组(C组)、生理盐水组(NS组)、曲马多组(T组)、神经病理性痛+生理盐水组(NP+NS组)和神经病理性痛+曲马多组(NP+T组).C组不行任何处理;NS组和T组仅暴露坐骨神经,分别腹腔注射生理盐水2 ml/kg或曲马多10 mg/kg;NP+NS组和NP+T组制备神经病理性痛模型,模型制备后第7天分别腹腔注射生理盐水2 ml/kg或曲马多10 mg/kg.除C组外,其余4组于腹腔注射曲马多或生理盐水前(T1)和注射后1 h(T2)时测定热痛阈和机械痛阈.于模型制备后第5天,左侧侧脑室注射30%霍乱毒素亚单位B与辣根过氧化物酶复合物(CB-HRP)3μl以标记远位触液神经元,并测定远位触液神经元5-HT1A表达水平.结果 与C组比较,NP+NS组中脑远位触液神经元5-HT1A受体表达下调(P＜0.05),其余组差异无统计学意义(P＞0.05);与NS组和T组比较,NP+NS组中脑远位触液神经元5-HT1A受体表达下调,热痛阈和机械痛阈降低,NP+T组热痛阈和机械痛阈降低(P＜0.05),中脑远位触液神经元5-HT1A受体表达差异无统计学意义(P＞0.05);与NP+NS组比较,NP+T组中脑远位触液神经元5-HT1A受体表达上调,热痛阈和机械痛阈升高(P＜0.05).结论 曲马多可下调中脑远位触液神经元5-HT1A受体的表达,该作用可能是其减轻大鼠神经病理性痛的机制之一.     

2015年乳腺癌辅助化疗进展

【摘要】〓乳腺癌是危害我国女性健康的头号杀手,尽管近年来辅助化疗的研究进展突飞猛进,但临床中仍有不少问题未能明确,如辅助化疗的合适人群、化疗的开始时间、蒽环及紫杉类的地位和用法、强化维持治疗的作用、疗效及预后的生物标志物等。本文结合乳腺癌辅助化疗在临床上的常见问题和2015年各大乳腺癌会议阐述乳腺癌辅助化疗的最新进展。     

Alterations in T-Cell Subset Frequency in Peripheral Blood in Obesity

Background: Obesity affects the regulation of immune and inflammatory responses. This study characterizes differences in peripheral blood lymphocyte phenotype in obese humans. Methods: Frequencies of lymphocyte subsets among peripheral blood mononuclear cells were compared between 10 obese (BMI ≥35) and 10 lean subjects, as determined by antibodies directed against cluster differentiation (CD) markers. Results: Obese patients demonstrated an increased frequency of CD3+CD4+ T-cells (mean difference 12%, P=0.004), a decreased frequency of CD3+CD8+ T-cells (mean difference 9.4%, P=0.016) and an increased frequency of CD3+CD8+CD95+ T-cells (mean difference 13.3%, P=0.032). No other differences among T-cell or monocyte subsets were noted. Conclusions: Obesity is associated with alterations in frequencies of peripheral CD4+ and CD8+ T-cells and aberrations in the expression of CD95 among CD8+ T-cells. These data suggest both CD4+ and CD8+ T-cell compartments, as well as the regulation of CD95 expression on CD8+ T-cells, as targets for further study into obesity's effects on the immune system.     

西宁地区烧伤病人动脉血气分析观察

对高海拔地区的27例烧伤病人动脉血气变化进行了分析和观察。结果证明:无论是存活病人还是死亡病人伤后均存在有低氧血症问题。并且在死亡病人和烧伤合并吸入性损伤病人其低氧血症的发生早于单纯烧伤病人。提示:吸入性损伤病人应立即行气管切开术以保障氧气供给,单纯烧伤病人可常规吸氧以维持正常血 PaO_2,ARDS 均发生在合并吸入性损伤的病人,高频喷射通气技术对纠正低氧血症有一定效果。     

Controversies in Fistula in Ano

Managing a complex fistula in ano can be a daunting task for most surgeons; largely due to the two major dreaded complications—recurrence & fecal incontinence. It is important to understand the anatomy of the anal sphincters & the aetiopathological process of the disease to provide better patient care. There are quite a few controversies associated with fistula in ano & its management, which compound the difficulty in treating fistula in ano. This article attempts to clear some of those major controversies.     

β-半乳糖苷酶在体内外成骨细胞中的表达

目的 研究β—半乳糖苷酶(β—gal)在成骨细胞中的表达状况，为阐明MorquioB综合征的发病机制提供依据。方法 裸鼠各器官和骨组织标本行X-gal染色检测。抽取羊和人骨髓行骨髓基质细胞(BMSCs)培养，分为4组：I：Adv-hBMP-2转染组；Ⅱ：Adv—β—gal转染组；Ⅲ：未转染组；Ⅳ：地塞米松诱导组。分别行X-gal染色和RT-PCR检测β—gal的表达。结果 裸鼠骺板两侧、骨膜内面及松质骨的成骨细胞和破骨细胞可见多量β—gal的表达。未转染BMSCs组有少量β—gal的表达，其他3组细胞的β—gal表达增高。结论成骨细胞和破骨细胞可表达多量β—gal，该两种细胞的β—gal缺乏可能是MorquioB综合征骨骼异常的直接原因。     

Smoking-Attributable mortality in Spain in 2016

IntroductionSmoking-attributable mortality (SAM) is a valuable indicator that can be used to characterize the course and health burden of the smoking epidemic. The aim of this paper was to estimate SAM in Spain in 2016 in the population aged 35 and over, using the best available evidence.MethodsA smoking prevalence-dependent analysis based on the estimation of population-attributable fractions was performed. Smoking prevalence (never, former, and current smokers) was calculated from a combination of the Spanish Health Survey (2016) and the European Health Survey (2014); the relative risk of death among current and former smokers was taken from the follow-up of various cohorts; and mortality rates were obtained from National Center for Statistics data. SAM estimates are presented globally, and by sex, age groups, and major disease categories: cancer, cardiometabolic diseases and respiratory diseases.ResultsIn 2016, 56,124 deaths were attributed to tobacco consumption, 84% in men (47,000), and 50% in the population aged over 74 (27,795). Overall, 50% of SAM was due to cancer (28,281), 65% of which was lung cancer. One in 4 attributable deaths (13,849) occurred before the age of 65.ConclusionsOne in 7 deaths in Spain in 2016 were attributable to smoking. This estimation of SAM clearly highlights the great impact of smoking on mortality in Spain, mainly due to lung cancer and chronic obstructive pulmonary disease.     

MicroRNAs in hepatic pathophysiology in diabetes

MicroRNAs(miRNAs or miRs) are small approximately 22 nucleotide RNA species that are believed to regulate diverse metabolic and physiological processes.In the recent past,several reports have surfaced that demonstrate the role of miRNAs in various biological processes and numerous disease states.For a disease as complex as diabetes,the emergence of miRNAs as key regulators leading to the disease phenotype has added a novel dimension to the area of diabetes research.On the other hand,the liver,a metabolic hub,contributes in a major way towards maintaining normal glucose levels in the body as it can both stimulate and inhibit hepatic glucose output.This equilibrium is frequently disturbed in diabetes and hence,the liver assumes special significance considering the correlation between altered hepatic physiology and diabetes.While the understanding of the mechanisms behind this altered hepatic behavior is not yet completely understood,recent reports on the status and role of miRNAs in the diabetic liver have further added to the complexities of the knowledge of hepatic pathophysiology in diabetes.Here,we bring together the various miRNAs that play a role in the altered hepatic behavior during diabetes.     

Fluid-phase transcytosis in the primate epididymis in vitro and in vivo

Ligated tubules from the corpus epididymidis of men and monkeys were incubated in medium containing horseradish peroxidase (HRP) as a marker for fluid-phase endocytosis. HRP was localized by light and electron microscopy after 0, 15, 30 and 60 min of incubation. Movement between the cells was prevented by tight junctions, but bypass of this barrier was apparently achieved by an intracellular vesicular mechanism leading to a time-dependent appearance of HRP in the lumen. Uptake of HRP into basal cells and capture by the lysosomal apparatus of principal cells were also observed. HRP-filled vesicles also appeared in the basal, mid and apical cytoplasm of epithelial cells in the caput 1 h after injection of the tracer into the epididymal circulation of the monkey, suggesting that this pathway also operates in vivo.