高温高湿环境颅脑火器伤后兔脑p38MAPK变化

OBJECTIVE: To study the activation of p38 mitogen-activated protein kinase (MAPK) in the brain of rabbits after craniocerebral gunshot injury in a hot and humid environment (HHE) and explore its possible mechanism. METHODS: Craniocerebral gunshot injury model was established in 30 New Zealand white rabbits, which were subsequently exposed to environment of normal temperature (at 22.0% +/- 0.5 degrees C; with relative humidity of 50%) or HHE at 39.0 +/- 0.5 degrees C; with relative humidity of 80%-85% for 10 min, 30 min, 1 h, 1.5 h, and 2 h groups, respectively, with 5 rabbits in each group. p38 MAPK activity in the brain tissues of the rabbits following the injury and environmental exposure were detected by Western blotting and analyzed semi-quantitatively by Bio-Profil gel image analysis system. RESULTS: p38 MAPK activity in the cortex and hypothalamus was significantly elevated following gunshot injury and HHE exposure, reaching the peak level at 1 h of HHE exposure and then decreased. p38 MAPK activity was significantly higher in the hypothalamus than in the cortex. CONCLUSION: p38 MAPK activity increases in the early stage following craniocerebral gunshot injury and HHE exposure in rabbits, the mechanism of which might involve the secondary brain insult.     

高温高湿环境颅脑火器伤后兔脑p38MAPK变化

目的 研究p38MAPK在高温高湿环境下颅脑火器伤后活化及其原因。方法 采用高温高湿颅脑枪伤模型,新西兰大白兔30只,随机分成常温[(22.0±0.5)℃、RH50%]对照组,高温高湿[(39.0±0.5)℃、RH80%~85%]枪伤后受热10和30min,1h、1.5h、2h组,每组5只。采用Westernblot检测脑匀浆p38MAPK活性,应用化学发光和X线片显示,凝胶图像分析仪半定量分析。结果 颅脑枪伤受高温高湿环境作用脑皮质和下丘脑的p38MAPK的活性受湿热后迅速增高,于受热1h达高峰,随后下降。下丘脑的p38MAPK活性比脑皮质高。结论 高温高湿环境颅脑火器伤后p38MAPK活性早期明显升高,参与了脑的继发性损害过程。     

高温高湿环境下犬贯通性颅脑火器伤模型的建立

目的 建立高温高湿环境下犬贯通性颅脑火器伤动物模型.方法 成年杂种犬40只,采用国产"54"式军用手枪,7.62mm手枪弹,枪口距犬额部10cm远处,冠状方向射击,制作颅脑贯通性火器伤模型.致伤后随机分为二组,即常温常湿组(对照组) 气温(Ta)(22.0±0.5)℃,相对湿度(RH)50%和高温高湿组(Ta) (39.0±0.5)℃,RH(80%～85%),每组20只.Powlab/8 sp生理记录仪动态监测体温、呼吸(R)、心率(HR)和平均动脉压(MAP)变化;监测血气分析和血电解质变化.结果 高温高湿环境下颅脑火器伤组伤情明显重于对照组,表现为生命体征紊乱,使呼吸明显加快,循环、体温调节功能衰竭;缩短了动物生存时间,增加动物的死亡率.结论 该动物模型对观察高温高湿环境下颅脑火器伤早期伤情变化特点是可行的、合理的,适应于高温高湿环境下颅脑火器伤的早期病理生理研究,并为该类创伤的早期救治提供实验依据.     

高温高湿环境下猫颅脑火器伤后细菌生长特点

OBJECTIVE: To investigate the growth behavior of characteristics in craniocerebral gunshot wound of cats in a hot and humid environment. METHODS: Twenty-three cross-bred cats were randomly divided into 4 groups: group A, the gunshot wound control group at normal temperature, in which tissue sampling was performed immediately after the wounding; group B, another gunshot wound control group at normal temperature, in which the samples were taken 6 h after the wounding; group C, the gunshot wound group subjected to a hot and humid environment, in which the tissue samples were obtained 6 h after the wounding; group D, the control group without undergoing the wound. The tissues from the wound tract and the surrounding tissues were sampled for bacterial culture and counting. RESULTS: The bacterial counts of the tissues from the wound tract, the tissues within 5 mm and within 5-10 mm from wound tract varied insignificantly between groups A, B and C (P>0.05). In each group, the bacterial counts declined in the tissues as the distance of the sampling sites from the wound tract increased (P<0.01). The bacterial counts of the tissues from the wound tract and within 5 mm from the wound tract in group A, B and C were significantly different from those in group D (P<0.01). CONCLUSION: Hot and humid environment does not significantly affect the bacterial growth in the craniocerebral gunshot wound within the first 6 h, which is a safe period against rapid bacterial growth and suitable for debridement.     

高温高湿环境兔颅脑火器伤后HSP70的变化

目的:研究热休克蛋白70(HSP70)在高温高湿环境下颅脑火器伤后的变化及其作用.方法:采用高温高湿颅脑枪伤模型,新西兰大白兔24只,随机分成常温对照组:(22.0±0.5)℃、RH 50%,常温枪伤组:(22.O±0.5)℃、RH 50%,高温高湿枪伤组(39.0±0.5)℃、RH 80%～85%,每组8只.采用Western blot法检测脑组织、淋巴细胞HSP70,应用化学发光和X线片显示,凝胶图像分析仪定量分析.结果:颅脑枪伤受高温高湿环境作用后,大脑皮层、下丘脑和淋巴细胞的HSP70显著增高;淋巴细胞中的HSP70与大脑皮层、下丘脑的变化趋势相一致.结论:脑组织和淋巴细胞中HSP70在高温高湿环境颅脑火器伤时显著增加.     

高温高湿环境下颅脑火器伤的基础与救治研究概论

较系统地介绍了高温高湿环境下颅脑火器伤的病理生理改变特点，指出高湿高湿环境是引起脑二次创伤的重要因素，是导致伤病员早期伤死的主要原因之一。提出加强这一军事医学课题研究、提高救治成功率的重要性。     

高温高湿环境肢体火器伤病理形态学变化

目的 通过建立高温高湿环境下犬肢体火器伤动物模型,探讨这一特殊环境下肢体火器伤的病理形态学变化,为其临床救治提供理论基础。方法 将８只杂种犬随机分为高温高湿组（５只）和常温常湿组（３只）,分别于火器伤后４、８、１２、２４h进行大体观察并通过光镜和电镜进行病理形态学观察。结果 大体观察可见高温高湿组６－８h后较常温常湿组道及肌肉变色区明显坟大,肢体肿胀加重,挫伤区色泽暗红,肌肉无收缩且有腐败臭味,有感染征;而常温常湿组１２－２４h伤道始出现臭味,光镜及电镜观察到高温高湿组各区肌肉纤维的病理变化均较常温常湿组显著,且损伤呈进行性加重,而常温常湿组的震荡区及震荡外区２４h伤道组织损伤均有减轻趋势。结论 高温高湿环境下肢体火器伤伤道肌肉组织病理损伤严重,且随时间的延长而逐渐加重,救治时尖强调早期彻底清创。     

高温高湿环境下颅脑火器伤的基础与救治研究概论

较系统地介绍了高温高湿环境下颅脑火器伤的病理生理改变特点,指出高湿高湿环境是引起脑二次创伤的重要因素,是导致伤病员早期伤死的主要原因之一。提出加强这一军事医学课题研究、提高救治成功率的重要性。     

高温高湿环境肢体火器伤病理形态学变化

目的通过建立高温高湿环境下犬肢体火器伤动物模型,探讨这一特殊环境下肢体火器伤的病理形态学变化,为 其临床救治提供理论基础。方法将８只杂种犬随机分为高温高湿组（５只）和常温常温组（３只）,分别于火器伤后４、８、 １２、２４ ｈ进行大体观察并通过光镜和电镜进行病理形态学观察。结果大体观察可见高温高湿组６－８ ｈ后较常温常湿组 伤道及肌肉变色区明显扩大,肢体肿胀加重,挫伤区色泽暗红,肌肉无收缩且有腐败臭味,有感染征象;而常温常湿组 １２～２４ ｈ伤道始出现臭味。光镜及电镜观察到高温高湿组各区肌肉纤维的病理变化均较常温常湿组显著,且损伤呈进行 性加重,而常温常温组的震荡区及震荡外区２４ｈ伤道组织损伤均有减轻趋势。结论高温高湿环境下肢体火器伤伤道 肌肉组织病理损伤严重,且随时间的延长而逐渐加重,救治时更应强调早期彻底清创。     

高温高湿环境下猫颅脑火器伤的病理学改变

目的 研究高温高湿环境下猫颅脑火器伤后病理学变化特点。方法 将杂种猫16只随机分为常温组、常温枪伤组、高温高湿组和高温高湿枪伤组，分别在光镜和电镜下观察其脑组织病理改变。结果 常温枪伤组早期表现为血管舒缩功能障碍，血管痉挛及扩张并存；高温高湿组血管痉挛减少，出血性表现增多；高温高湿枪伤组脑组织切片及超薄切片观察发现神经细胞存活数量明显减少，细胞器水肿变性，毛细血管紧密连接增宽，毛细血管破裂出血，髓鞘变性松解等病理变化，较常温颅脑火器伤中明显加重。结论 高温高湿环境对颅脑火器伤时的病理变化影响显著。     

p38丝裂原活化蛋白激酶与高血压血管重构

高血压时,血管平滑肌细胞的增殖、细胞外基质的改变、内皮细胞损伤等因素均可引起血管的重构。p38丝裂原活化蛋白激酶主要介导细胞的发生、分化、增殖、凋亡等多种病理生理过程。本文简要介绍了p38丝裂原活化蛋白激酶信号转导通路与高血压血管重构的关系。     

p38蛋白激酶(p38 MAPK)在癫疒间大鼠脑内的表达

 目的  观察p38蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)在癫疒间大鼠脑内的表达情况。方法  健康雄性SD大鼠随机分成正常对照组(n=8)和癫疒间组(n=8)。采用戊四氮腹腔注射建立癫疒间模型,大鼠点燃后的惊厥行为按照Racine的标准进行观察评分,采用Western blot和免疫荧光法比较两组大鼠脑内p38 MAPK的表达情况。结果  癫疒间组大鼠脑内p38 MAPK在皮层和海马的表达均显著高于正常对照组(P<0.01)。结论  p38 MAPK在癫疒间大鼠脑内表达上调。     

基于细胞穿透肽技术对p38丝裂原活化蛋白激酶蛋白功能的研究

目的 构建基于TAT技术的p38 MAPK蛋白转运系统并对导入细胞的融合蛋白的功能进行鉴定。方法 采用亚克隆方法构建重组质粒pHis-TAT-p38和pHis-TAT-p38（AF）无活性突变体,并诱导原核表达纯化融合蛋白;将这两种蛋白加入培养的ECV304细胞,在高渗刺激下,通过检测p38底物ATF2的磷酸化水平,以观察由TAT转导进入细胞His-TAT-p38及其无活性突变体对内源性p38活性的影响。结果 酶切和测序结果表明,载体构建正确;SDS-PAGE凝胶电泳可见原核表达纯化得到高纯度目的蛋白：Western blot表明,融合蛋白His-TAT-p38及其突变体能够以一种时间和浓度依赖性方式高效转导进入细胞：导入His-TAT-p38蛋白后,结果发现导入的野生型p38可增强内源性p38的功能,而无活性突变体则竞争性抑制了内源性p38MAPK蛋白的功能,从而阻止或抑制其对内源性底物ATF2的磷酸化．使得信号不能传递下去,进而抑制p38MAPK通路的活动。结论 成功建立了基于TAT的蛋白转运系统．并证实TAT能够以浓度和时间依赖方式高效转运蛋白进入真核细胞;进入细胞的His-TAT-p38和His-TAT-p38无活性突变体蛋白均具有较高的生物学活性,在高渗刺激作用下前者可以增加p38磷酸化水平,而后者则显著抑制p38信号通路的活性。     

高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶的影响

目的 探讨高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶表达的影响.方法 幼年Wistar大鼠90%氧气暴露建立高氧肺损伤模型,行肺组织病理学检查,应用TUNEL法检测肺组织的细胞凋亡,Western blot法检测p38 MAPK表达.结果 高氧暴露3 d可见肺组织水肿、出血、炎症细胞浸润等急性肺损伤改变.高氧3 d组的肺凋亡指数较空气对照组明显增加,凋亡发生的主要部位为肺泡、支气管上皮及血管内皮细胞.Westem blot结果显示高氧暴露1 d,肺组织的p38MAPK活性迅速升高,于第2天达到高峰,第3天活性有所下降,但仍高于空气对照组.结论 高氧暴露可以诱导肺组织发生细胞凋亡;高氧可以激活p38MAPK通路,参与高氧性肺损伤的调控;活化的p38MAPK可能参与了高氧诱导的肺组织细胞凋亡的调控.     

The Relationship between p38MAPK and Apoptosis during Paclitaxel Resistance of Ovarian Cancer Cells

To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC50) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and West- ern blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7±1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to pacli- taxel was (57.18±2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to pacli- taxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in pa- clitaxel-resistant ovarian carcinoma cells depends on the activation of p53.     

The Relationship between p38MAPK and Apoptosis during Paclitaxei Resistance of Ovarian Cancer Cells

To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC(50)) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and Western blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7+/-1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to paclitaxel was (57.18+/-2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to paclitaxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in paclitaxel-resistant ovarian carcinoma cells depends on the activation of p53.     

The relationship between p38MAPK and apoptosis during paclitaxel resistance of ovarian cancer cells

Summary To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC₅₀) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and Western blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7±1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to paclitaxel was (57.18±2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to paclitaxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in paclitaxel-resistant ovarian carcinoma cells depends on the activation of p53. This project was supported by a grant from R&D program of Heilongjiang Province (No. GB05C402-11).     

坐骨神经压迫性损伤大鼠背根神经节磷酸化p38MAPK表达的变化

目的:观察背根神经节(DRG)磷酸化的p38丝裂原活化蛋白激酶(p -p38MAPK)在大鼠坐骨神经慢性压迫性损伤(CCI)后表达的变化,探讨慢性神经痛的发生机制。方法:2 4只雄性SD大鼠随机分成空白对照组(Naive组)、假手术组(Sham组)、坐骨神经结扎后7d组和1 4d组(CCI7d和CCI1 4d组) ,分别在各自的时间点灌注取材L4- 5背根神经节,用免疫组织化学方法观察磷酸化p38MAPK表达的变化。结果:坐骨神经结扎组背根神经节磷酸化的p38MAPK免疫阳性神经元数和染色深度均明显增加,与假手术组或空白对照组相比,差异具有显著性(P <0 .0 1 )。结论:坐骨神经损伤后背根神经节p38MAPK的激活与神经性疼痛的病理过程密切相关。