大剂量甲基强地松龙对大鼠脊髓损伤后iNOS表达及细胞凋亡的影响

目的：探讨大鼠脊髓损伤后应用大剂量甲基强地松龙(MP)对诱导型一氧化氮合酶(iNOS)表达及细胞凋亡的影响。方法：成年大鼠随机分为脊髓损伤后应用大剂量甲基强的松龙治疗组(A组)和应用生理盐水对照组(B组)，损伤后不同时间点(4h、8h、1d、3d、7d、14d、21d)按Tarlov标准评价大鼠双后肢神经功能恢复情况，然后处死。用HE染色观察损伤脊髓组织病理变化，用免疫组化染色检测iNOS阳性细胞，原位末端标记法(TUNEL法)标记凋亡细胞。结果：HE染色镜检发现脊髓组织病理学改变A组明显轻于B组。A、B两组均发现凋亡细胞及iNOS表达，神经细胞凋亡指数及iNOS表达均为B组＞A组(P＜0．01)。结论：大剂量甲基强的松龙能抑制大鼠脊髓损伤后iNOS表达及细胞凋亡。     

神经营养素3对大鼠脊髓损伤后运动功能恢复的影响

目的 探讨神经营养素3(neurotrophin-3,NT-3)对大鼠急性脊髓损伤的治疗作用.方法 实验于2005-01/06在解放军第三军医大学中心实验室完成.选择60只健康SD大鼠,随机分为3组:对照组(生理盐水组),实验组(NT-3组),假手术组(n=5);用改良Allen's法以30gcm致伤SD大鼠制作大鼠T8全瘫模型,实验组经蛛网膜下腔导管于术后即刻,4h,8h,12h,24h,3d,7d注入NT-320μl(含NT-3 200ng),对照组在相同时间点给予等量生理盐水,假手术组只打开椎板后蛛网膜下腔置管,不致伤,不给药.各组动物分别于术后24h,3d,7d、14d行BBB评分和MEP检查.结果 60只大鼠全部进入结果分析,①实验组在24h、3d、7d、14d较对照组BBB评分分值提高[0.22±0.43/0.70±0.48(P＜0.05);3.2 2.39/8.01 2.05;9.30 1.49/14.32 2.11;12.80±0.92/18.40±1.08(P＜0.01)],②实验组MEP的N1波潜伏期在24h、3d、7d、14d均较对照组明显缩短[80.90±7.49/49.9±2.44(P＜0.01);54.05±8.52/40.55±4.72(P＜0.05);43.10±1.53/33.65±2.78(P＜0.01);38.45±3.98/22.25±2.84(P＜0.01)].结论 神经营养素3能明显促进脊髓损伤后运动功能的恢复,对大鼠脊髓损伤有明显治疗作用.     

氯胺酮对慢性坐骨神经压迫性损伤大鼠脊髓背角神经元凋亡及Bax和Bcl-2的表达影响

目的观察氯胺酮（Ket）对慢性坐骨神经压迫性损伤（CCI）大鼠脊髓背角神经元凋亡及凋亡相关蛋白Bax和Bcl-2表达的影响。方法选择雄性SD大鼠40只，随机分为4组，分别为空白对照组，假手术组，CCI组和腹腔注射Ket 50mg／kg治疗组。假手术组，CCI组和Ket治疗组又按术后取材时间的不同分别分为3个亚组，即术后3d组、术后9d组和术后14d组。用TUNEL标记法标记脊髓背角浅层凋亡细胞。应用免疫组织化学方法和免疫印迹（Western-blot）法检测大鼠脊髓背角浅层神经元凋亡相关蛋白Bax和Bcl-2表达情况。结果与空白对照组和假手术组各亚组比较，CCI3d，9d，14d亚组和Ket9d，14d亚组大鼠脊髓背角的凋亡细胞明显增多（P〈0．01）；与CCI3d亚组比较，Ket3d亚组大鼠脊髓背角的凋亡细胞明显减少（P〈0．01）。与空白对照组和假手术各亚组比较，CCI3d，9d，14d亚组大鼠脊髓背角浅层Bax阳性神经元和Bax的表达显著增加（P〈0．01），CCI9d，14d亚组大鼠脊髓背角浅层Bcl-2阳性神经元和Bcl-2的表达明显增加（P〈0．01）。与CCI3d亚组比较，Ket3d亚组脊髓背角浅层Bax阳性神经元和Bax的表达显著减少（P〈0．01），而Bcl-2阳性神经元和Bax的表达显著增加（P〈0．01）。结论Ket能抑制脊髓背角浅层神经元Bax的表达，促进Bcl-2的表达，提示Ket对神经病理性疼痛状态下脊髓背角神经元的凋亡具有保护作用。     

复方丹参对大鼠脊髓损伤后细胞凋亡及诱导型一氧化氮合酶表达的影响

目的:探讨大鼠脊髓损伤后应用复方丹参对细胞凋亡及诱导型一氧化氮合酶(iNOS)表达的影响。方法:成年大鼠随机分为正常组、脊髓损伤后应用复方丹参组(组)和应用生理盐水对照组(组),损伤后不同AB时间点(8h、1d、3d、7d、14d、28d)处死大鼠,用HE染色观察损伤脊髓组织病理变化,原位末端标记法(TUNEL法)标记凋亡细胞,用免疫组化染色检测iNOS阳性细胞。结果:HE染色镜检发现脊髓组织病理学改变A组明显轻于B组。A、B两组均发现凋亡细胞及iNOS表达,神经细胞凋亡指数及iNOS表达均为B组>A组(P<0.01)。结论:复方丹参能抑制大鼠脊髓损伤后细胞凋亡及iNOS表达。     

骨髓间充质干细胞移植促进脊髓损伤大鼠神经功能的恢复

目的 探讨骨髓间充质干细胞(MSCs)移植对脊髓损伤大鼠神经功能恢复的影响极其可能机制.方法 采集SD大鼠骨髓,分离出MSCs,并培养、纯化,取传至第6代的MSCs,移植前1 d以5溴2脱氧尿嘧啶核苷(Brdu)标记,然后用显微注射器将其缓慢注射到脊髓损伤大鼠模型(成年SD大鼠)的受损脊髓中心,以不进行移植的脊髓损伤大鼠模型(损伤组)和假手术组为对照.术后进行运动功能评分以判断神经功能恢复情况,并切取移植区域脊髓组织,应用荧光免疫化学染色检测脊髓损伤灶边周的神经元凋亡情况(TUNEI.与NeuN双标记)以及移植的MSCs的分布和向神经元分化情况(Brdu与NeuN双标记),应用逆转录聚合酶链反应检测损伤灶区域脑源性神经营养因子(BDNF)mRNA的表达.结果 移植后第3天开始,损伤组和移植组大鼠的BBB运动功能评分明显上升,第14天后进入平台期,但移植组各时间点的.BBB运动功能评分均明显高于损伤组(P<0.05).移植后3 d,在移植组的脊髓组织中可见Brdu标记阳性细胞,少部分Brdu标记阳性细胞同时表达神经元特异性标志物NeuN;移植后第3、7天,损伤组和移植组脊髓组织中的凋亡神经元均显著多于假手术组(P<0.01),凋亡的神经元多存在于损伤周边区,但移植组的凋亡神经元显著少于损伤组(P<0.01);移植后第3、7天,假手术组未检测到BDNF mRNA的表达,损伤组和移植组均可检测到BDNF mRNA的表达,移植组的表达量分别较损伤组高28.6%和39.2%(P<0.05).结论 移植的骨髓MSCs可促进脊髓损伤大鼠神经功能的恢复,其机制除MSCs直接分化为神经元进行修复外,还可能与其改善脊髓损伤区的微环境、上调BDNF、mRNA表达、减少神经细胞凋亡有关.     

Ephrin-B1在大鼠脊髓损伤的表达

[目的]探讨红细胞生成素诱导肝癌细胞株配体-B1 (Ephrin-B1)在大鼠脊髓损伤的表达变化.[方法]将60只成年Wistar大鼠分正常组、假手术组、伤后3、7、14 d及28 d组.假手术组大鼠不损伤脊髓,损伤组大鼠采用Allen's氏打击法损伤T10脊髓.分别在各个时间点取材,免疫荧光和Western Blot检测Ephrin-B1蛋白表达情况.[结果]免疫荧光结果显示Ephrin-B1在正常、假手术和损伤脊髓的灰质和白质中均有表达,但在损伤脊髓组织中表达较高,脊髓组织中的星形胶质细胞表达Ephrin-B1.Western Blot显示损伤组比对照组Ephrin-B1蛋白含量显著增加(P＜0.01),从伤后3d开始持续到伤后28 d;伤后14 d蛋白含量达到高峰,明显高于对照组及伤后其他组(P＜0.01);伤后28 d蛋白含量较14 d减少(P＜0.01),与伤后7d相比无显著性差异(P＞0.05).[结论]Ephrin-B1在大鼠脊髓损伤后表达升高,伤后14d表达升高显著,表达于脊髓的星形胶质细胞.     

红花黄素腹腔注射联合BMSCs移植对大鼠脊髓损伤的保护作用

目的 探讨红花黄素腹腔注射联合骨髓阳J充质干细胞(BMSCs)移植对大鼠脊髓损伤保护作用及其机制.方法 体外分离、培养BMSCs;SD大鼠45只,改良Allen法制备脊髓损伤模型,随机分为3组:A组、B组、C组.A组腹腔注射红花黄素治疗(40 mg/kg),B组和C组注射等体积的生理盐水,每日1次,共14 d;A组和B组损伤局部行BMSCs移植(6×106/60μl),C组注射等体积的PBS液.于术后1 d、2 d、3 d、4 d、7 d取材检测损伤处超氧化物歧化酶(SOD)、丙二醛(MDA)和细胞凋亡,术后1 d和7 d、14 d、21 d、28 d采用改良BBB评分法进行行为学检查评分.结果 术后随着时间的推移,各组BBB评分得分均增高,术后28 d时A组得分为(19.33±1.37),B组为(15.83±1.17),C组为(9.0±1.41),A组高于B组和C组,B组高于C组,差异有统计学意义(P＜0.05).SOD活性在相同时间点,均有A组高于B组和C组,B组高于C组,差异有统计学意义(P＜0.05);MDA含量在术后2 d、3 d、4d A组低于B组和C组(P＜0.05),且有A组低于B组(P＜0.05).术后3 d时各组细胞凋亡达高峰:A组(72.83±13.38),B组(96.33±6.35),C组(219.33±28.64);之后各组细胞凋亡数逐渐减少,至术后7d:A组(3.67±3.72),B组(16.33±5.54),C组(86.17±12.27);除术后第1天,在相同时间点,细胞凋亡数有A组少于B组和C组,B组少于C组,差异有统计学意义(P＜0.05).结论 红花黄素和BM-SCs移植均能抑制损伤脊髓局部自由基生成、脂质过氧化和细胞凋亡,减轻继发损伤,两者联合具有累加协同效应,有助于脊髓损伤大鼠肢体运动功能的恢复.     

牙髓干细胞移植治疗大鼠脊髓损伤的实验研究

目的:探讨牙髓干细胞移植治疗脊髓损伤的神经修复及运动功能恢复情况.方法:培养取材第2代来源于第三磨牙的人牙髓于细胞,鉴定其表面标记物,应用B27、碱性呈纤维细胞生长因子和胰岛素转铁蛋白硒进行神经诱导,并行免疫荧光染色检测.改良Allen's法制作SD大鼠脊髓损伤模型,3d后随机分为实验组及对照组,每组各20只大鼠.分别于脊髓损伤处注入人牙髓干细胞及生理盐水,于造模后1d、治疗前1d、治疗后3d、治疗后7、14、28d进行动物后肢运动功能检测.28d时脊髓取材进行HE染色,观察脊髓空洞形成,计算空洞面积;Tunnel法检测两组细胞凋亡情况;免疫荧光双标法标记HuNu-NeuN和HuNu-GFAP,观察人牙髓干细胞体内生长分化情况.结果:细胞培养传代后呈长梭形,细胞形态均匀,胞浆丰富、胞核增大,细胞平行排列呈漩涡状或螺旋状.流式细胞仪分析人牙髓干细胞诱导分化后高表达CD44、CD90和CD146,低表达STRO-1,CD34、CD45表达阴性.人牙髓干细胞神经诱导14d,免疫荧光标记GFAP、NeuN呈阳性.细胞移植3d、7d,两组间BBB评分无统计学差异;细胞移植14d及28d实验组BBB评分分别为3.8±0.8、7.2±1.6,对照组BBB评分为2.2±0.8、3.6±1.1,两组间比较差异有显著性(P＜0.05).28d时HE染色两组均可见脊髓出血、炎性细胞浸润、小血管增生及空洞形成.实验组脊髓空洞面积百分比(26.75±2.50)％,对照组为(49.50±6.25)％,两组差异有统计学意义(P＜0.05);Tunnel法检测实验组神经细胞凋亡百分比(32.33±1.54)％,对照组为(46.33±1.53)％,实验组显著减少神经细胞的凋亡(P＜0.05).免疫荧光双标法检测到部分细胞为带有HuNu-NeuN和HuNu-GFAP双抗体细胞.结论:人牙髓干细胞能够在体外特定条件下及移植入脊髓损伤大鼠体内可分化为神经细胞,用于治疗脊髓损伤时可减少神经细胞的凋亡,促进后肢运动功能的恢复.     

不同年龄大鼠颈上神经节节前神经元在脊髓内分布差异及其机制

目的 观察不同年龄大鼠颈上神经节节前神经元在脊髓内的分布及其传出纤维与臂丛神经的关系,为阐明Horner征在产瘫手术指征及预后判断中的价值提供实验依据.方法 取出生后7、14 d及成年SD大鼠各24只,各年龄段随机分为保留C7、保留C8、保留T1和保留T2组.将FG注入大鼠双侧颈上神经节,48～72 h后计数脊髓内FG标记神经元,并行TUNEL神经元凋亡检测,计算神经元凋亡率.结果 在保留C7组,生后7 d大鼠FG标记神经元数为240±72,占总数的(15.64±4.82)%;14 d大鼠为191±64,占(13.64±6.95)%;成年大鼠仅为11±5,占(0.91±0.45)%,幼年大鼠(生后7 d和14 d)FG标记神经元比例明显高于成年大鼠(P＜0.01).其余各组不同年龄大鼠间差异无统计学意义.幼年大鼠FG标记神经元保留C7组凋亡率最高,在7 d和14d大鼠中分别为为(3.1±0.9)%和(1.9±0.8)%,成年大鼠未检测到凋亡神经元.结论 幼年大鼠有一部分交感神经节前神经元发出的节前纤维通过C7神经根传出支配头面部,但成年后该部分神经元凋亡.     

脊髓损伤大鼠触液神经元中p75NTR的表达

目的 :探讨大鼠触液神经元(cerebrospinal fluid-contacting neurons,CSF-CNs)p75神经营养因子受体(p75 neurotrophin receptor,p75NTR)在脊髓损伤后的表达变化。方法 :成年雌性Sprague-Dawley(SD)大鼠36只,按随机数字表法分为正常对照组(6只)、假手术组(6只)和脊髓损伤组(24只),脊髓损伤组采用Allen′s打击模型(10g×3cm)在大鼠脊髓T10段造成急性脊髓损伤,分别于损伤3d、1周、2周、4周后进行取材;对照组不做任何处理,假手术组只暴露脊髓,不击伤脊髓。对各组大鼠运动功能行BBB评分,各时间点取材行病理切片HE染色观察。取材前48h侧脑室注射霍乱毒素B亚单位与辣根过氧化物酶复合物(CB-HRP)特异性标记触液神经元。处死大鼠后,取损伤的脊髓节段10mm,用免疫荧光双标法检测触液神经元p75NTR的表达,ImagePro Plus计数目标神经元CB-HRP/p75双阳性细胞的数目。结果 :假手术组各时间点BBB评分均为21.0±0;脊髓损伤组在术后3d、1周、2周、4周各时间点BBB评分分别为3.20±0.81、10.73±1.02、12.48±1.86、13.29±1.93,两组各时间点差异均具有统计学意义(P0.05)。HE染色可见正常对照组和假手术组脊髓组织结构完整,细胞形态正常;脊髓损伤组脊髓组织结构紊乱,神经元变性坏死,胶质细胞增生,胶质瘢痕和脊髓空洞形成。免疫荧光双标示正常对照组和假手术组可见少量CB-HRP/p75双阳性细胞,计数分别为5.16±0.55、4.31±0.61,两组比较差异无统计学意义(P0.05);脊髓损伤组伤后3d、1周、2周CB-HRP/p75双阳性细胞数分别为13.35±1.53、21.68±2.15、16.26±2.09,与正常对照组和假手术组比较差异均有统计学意义(P0.05),伤后4周时,CB-HRP/p75双阳性细胞数为4.83±0.73,与正常对照组和假手术组比较差异无统计学意义(P0.05)。结论 :p75NTR可在大鼠脊髓触液神经元中表达,且在脊髓损伤后表达增加,触液神经元可能通过p75NTR参与脊髓损伤的修复过程。     

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。     

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.     

Surgical management of ductal carcinoma in situ in Australia in 1995

Background: In the present paper we describe the presentation and management of ductal carcinoma in situ (DCIS) of the breast in women in Australia in 1995. This representative, national data set provides a historical comparator for studies examining DCIS management that follow. Methods: Surgeons identified by population‐based cancer registries as having treated a new diagnosis of DCIS between 1 April and 30 September 1995 completed a questionnaire on the presentation and management of each case. Results: Two hundred and five surgeons supplied treatment details on 418 DCIS tumours in 415 women . Half of all tumours were detected at BreastScreen clinics and a further 25% were detected at other mammography centres. Twenty‐six percent of tumours were palpable at presentation, 33% were multifocal and 55% were high grade (including comedocarcinoma). Breast conserving therapy (BCT) rather than mastectomy was utilized in 260 (62%) of cases. Tumours that were of low grade, small in size and not multifocal were more likely to be treated by BCT. Surgeons seeing six or more DCIS cases in the 6‐month period were more likely to utilize BCT. Of the conservatively treated cases, 22% were referred for a radiation oncology consultation. The most common reasons for treating DCIS with mastectomy were that the tumour was too extensive or multifocal (63%), it extended to margins of the specimen (42%), or patient concerns about recurrence (34%). Conclusions: In 1995 the majority of DCIS was treated with breast conserving surgery alone. Surgeons treating more DCIS cases were more likely to perform conservative surgery than surgeons treating only one DCIS case in the study period.     

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.