Study on the Regulation of Bcl-2 Gene on Rat Spermatogenic Cells Apoptosis in Transcription Level

Objective To detect the change of Bcl 2 gene expression in the apopototic process of spermatogenic cells in rat with vasoligation and vasostomy, and to find out the relationship between the transcription of Bcl 2 and the apoptosis of spermatognic cells Materials & Methods Sixty adult male Sprague Dawley rats in 3 groups were operated with vasoligation and vasostomy. Then hybridization in situ with hypersensitive Bcl 2 RNA probe was used to detect the change of Bcl 2 mRNA. Results The transcription of Bcl 2 gene in spermatogenic cells was obviously inhibited in the vasoligation group compared with that in the control group (P<0.05), and the transcription in the vasostomy group showed no difference from that of the control group. Conclusion Bcl 2 gene has an anti apoptotic effect in rats with vasostomy, and there was a transcriptional regulation of Bcl 2 gene in rat spermatogenic cell during the period of pre vasoligation to post vasoligation and to post vasosotomy.

Effect of basic fibroblast growth factor and danshen on bcl-2 and p53 mRNA expression in the brain of rats exposed to repeated, high, positive acceleration (+Gz)

BACKGROUND： Both animal experiments and clinical studies have shown that basic fibroblast growth factor （bFGF） and danshen （Salvia miltiorrhiza） can exhibit protective effects on ischemia-reperfusion cerebral injury. OBJECTIVE： To test whether bFGF and danshen can protect cerebral injury induced by exposure to repeated, high, positive acceleration （＋Gz） in an animal model and to analyze the possible mechanisms. DESIGN, TIME AND SETTING： Randomized controlled animal study. The experiment was performed at the Research Center for Molecular Biology, Air-force General Hospital of Chinese PLA from April to August 2000. MATERIALS： A total of 20 clean grade, healthy, Sprague Dawley rats of both genders, weighing （200 ± 15） g, were provided by our experimental animal center. Rats were randomly divided into 5 groups： the control group, ＋Gz exposure group, bFGF group, danshen group, and saline group, with 4 animals per group. bFGF （Beijing Bailuyuan Biotechnology Co. Ltd.） and danshen solution （Shanghai Zhongxi Pharmaceutical Co. Ltd.） were used. METHODS： All rats were fixed on a rotary arm of a centrifugal apparatus （2 m in radius） with their heads oriented towards the center of the apparatus. Except for rats in the control group, the value of ＋Gz exposure was ＋14 Gz with an acceleration rate of 1.5 G/s. The peak force lasted for 45 seconds. ＋Gz exposure was performed three times with intervals of 30 minutes. Rats in the control group received the same ＋Gz procedure, but the G value was ＋1 Gz. Rats in bFGF group and danshen group were intraperitoneally injected with 100 μg/kg bFGF or 15 g/kg danshen solution, respectively, at 30 minutes prior to centrifugation and immediately after centrifugation. Rats in saline group were injected with the same volume of saline. Six hours after exposure, rats were decapitated. One hemisphere was preserved in liquid nitrogen for RNA extraction and the other was processed for apoptosis detection. MAIN OUTCOME MEASURES： mRNA levels of bcl-2 and p53 were measured by semi-quantitative reverse-transcription polymerase chain reaction. Apoptotic cell death was detected by terminal deoxynuleotidyl transferase-mediated dUTP nick end labeling. RESULTS： Changes in mRNA expression of bcl-2 and p53 and apoptotic cells were observed in rat brain six hours after repeated ＋Gz exposures, bFGF and danshen were able block the changes of bcl-2 and p53 expression and inhibit apoptotic cell death. CONCLUSION： The data suggest that apoptosis and changes in bcl-2 and p53 expression in the rat brain can be induced by repeated ＋Gz exposures. Apoptosis is, therefore, one of the molecular mechanisms of brain damage induced by repeated ＋Gz exposures, bFGF and danshen were of the equal potency in preventing brain injury induced by repeated ＋Gz exposures.

THE EXPRESSION OF BCL-2 GENE AFTER TRANSIENT FOCAL ISCHAEMIA AND THE EFFECT OF MK-801

in order to clarify the pesible role of the bcl-2 gene imolved in the cell death Program,and the relatiouship of glutamate receptors with bcl-2 gene expressin, this study examied the expression of bcl-2 gene protein, the neuronal status of apoptosis and the effects of MK-801 using immunohistochemistry and in situ terminal.labelling methods after 30 min of.middle cerebral artery(MCA) occlusion and followed by 24 h of reperfusion. The presence of bcl-2 gene protein increased in the ipeilateral hemisphere of ischaemis espeially in the MCA territory MK-801 enhanced the expresion of the bcl-2 gene protein. No DNA fragmentation was detected in this experiment. In conclusion. bcl-2 gene activity increased during transient focal ischaemia, and was potentiated by MK MK801, which may be an endogenous protective mechanism .against ischaemic apoptosis. Apoptosis wasnot detected after tranient focal ischoemia. for 30 min rollowed by 24 h of reperfusiou.

醒脑静对急性重度CO中毒大鼠神经元Bcl-2基因表达的影响

目的研究醒脑静注射液(xingnaojing injection,XNJI)对急性重度CO中毒大鼠海马神经元Bcl-2基因表达和迟发性损伤的影响。方法45只雄性SD大鼠随机分成3组:正常对照组、CO中毒组、XNJI治疗组。各组大鼠在治疗完毕后取脑组织分别行常规HE染色、免疫组化染色和TUNEL染色,观察CO染毒后1、3、5和7 d大鼠海马神经元损伤和Bcl-2基因表达变化的特点。结果CO中毒组大鼠海马发生迟发性神经元凋亡,Bcl-2表达无明显变化;XNJI治疗组凋亡神经元明显减少,Bcl-2蛋白表达增多,尤以染毒后3 d和5 d明显(P<0.05)。结论XNJI可以促进急性CO中毒大鼠海马神经元Bcl-2表达,抑制神经元凋亡,具有保护神经元的作用。

不同月龄大鼠阴茎组织中血红素氧合酶2和一氧化碳含量的比较研究

目的:比较不同月龄大鼠阴茎海绵体组织中血红素氧合酶2(HO-2)及一氧化碳(CO)的含量,探讨HO-2/CO系统与年龄的相关性。方法:应用SABC免疫组化染色法观察8、16、24月龄(每组10只)大鼠阴茎组织中的HO-2并用图像分析法测定其含量;通过碳氧血红素曲线法测定阴茎组织中CO的含量。结果:阴茎海绵体组织中普遍存在HO-2,特别是在阴茎动脉外膜层周围及血窦内皮细胞系统中的含量较多,并且随着月龄的增长,阴茎海绵体组织中HO-2的含量逐步下降,高月龄组(16、24月龄)大鼠阴茎海绵体组织中HO-2含量明显低于低月龄组(8月龄)大鼠(P<0.05);阴茎海绵体组织中CO水平也随着大鼠月龄的增长显著降低(P<0.05)。结论:阴茎海绵体组织中因HO-2含量降低而导致CO含量下降且与年龄增长相关。

愈肾合剂对糖尿病模型大鼠肾组织血红素氧合酶-1和血红素氧合酶-2表达的影响

目的观察愈肾合剂对糖尿病模型大鼠肾脏组织血红素氧合酶(HO)-1、HO-2的影响,探讨其治疗糖尿病肾病作用机制。方法 SD大鼠随机分为正常组、模型组、对照组、治疗组。采用皮下注射链脲佐菌素法复制模型,对照组和治疗组给予相应药物灌胃,正常组和模型组灌胃等量蒸馏水。给药8周后,收集尿液测定24 h尿蛋白(Upro)、尿肌酐(Ucr),断头取血测定一氧化碳(CO)、尿素氮(BUN)、血肌酐(Scr),采用免疫组织化学法检测肾脏组织HO-1、HO-2的表达。结果与正常组比较,模型组大鼠BUN、24 h Upro、Scr升高,肌酐清除值(Ccr)、血浆CO降低,肾脏组织的HO-1、HO-2表达明显减弱(P<0.01);与模型组比较,治疗组和对照组BUN、24 h Upro、Scr明显下降,Ccr、血浆CO活性升高,肾脏组织的HO-1、HO-2表达明显增强(P<0.05,P<0.01);与对照组比较,治疗组24 h Upro降低,血浆CO活性升高,肾脏组织HO-1、HO-2表达明显增强(P<0.05,P<0.01)。结论愈肾合剂对糖尿病肾病大鼠有一定的治疗作用,可能是通过HO/CO系统实现的。

血红素氧合酶-2在新生大鼠延髓的表达及意义

目的探讨产生内源性一氧化碳(carbon monoxide,CO)的关键酶血红素氧合酶-2(heme oxygenase-2,HO-2)在新生大鼠延髓的表达及意义。方法采用新生Sprague-Dawley大鼠,行石蜡切片,分别进行HE染色和免疫组织化学染色(SP法),观察新生大鼠延髓神经核团的分布和不同神经核团中神经元的形态,以及新生大鼠延髓是否存在HO-2阳性神经元和阳性神经元的分布情况。结果在舌下神经核、疑核、巨细胞网状核、面神经核等部位均可见HO-2阳性神经元。结论 HO-2阳性神经元存在于新生大鼠延髓呼吸相关神经核团,提示内源性CO可能参与中枢性呼吸活动的调节。

急性一氧化碳中毒患者血清Tau蛋白、心型脂肪酸结合蛋白、可溶性生长刺激表达基因2蛋白与病情严重程度和预后的关系

目的探讨急性一氧化碳中毒(ACMP)患者血清Tau蛋白、心型脂肪酸结合蛋白(H-FABP)、可溶性生长刺激表达基因2蛋白(sST2)与病情严重程度和预后的关系。方法回顾性分析2019年1月至2021年6月西安市中心医院职业病科收治的50例ACMP患者的临床资料,按照中毒程度分为轻度组16例、中度组23例和重度组11例,检测并比较三组患者的血清Tau蛋白、H-FABP、sST2水平。此外,根据脑功能分级(CPC)评分的不同将患者分为预后不良组23例和预后良好组27例,比较两组患者的血清Tau蛋白、H-FABP、sST2水平。采用多因素Logistic回归分析建立多指标联合应用预测ACMP患者预后的回归预测模型。以受试者工作特征曲线(ROC)分析血清Tau蛋白、H-FABP、sST2预测ACMP患者预后的效能。结果不同中毒程度患者血清Tau蛋白、H-FABP、sST2水平比较,重度组[(68.84±16.84)pg/mL、(50.34±12.37)μg/L、(42.41±2.56)μg/L]明显高于中度组[(45.86±11.93)pg/mL、(28.48±6.04)μg/L、(37.56±2.03)μg/L],中度组明显高于轻度组[(22.03±6.23)pg/mL、(19.21±2.55)μg/L、(33.52±1.63)μg/L],差异均有统计学意义(P<0.05);不同预后患者血清Tau蛋白、H-FABP、sST2水平比较,预后不良组[(50.31±13.29)pg/mL、(35.20±10.05)μg/L、(40.38±2.16)μg/L]明显高于预后良好组[(37.31±7.94)pg/mL、(26.17±4.57)μg/L、(34.74±1.26)μg/L],差异均有统计学意义(P<0.05);预后不良组患者的年龄大于预后良好组,中毒时间长于预后良好组,急性生理学及慢性健康状况评分系统(APACHEⅡ)评分、肌酸激酶水平高于预后良好组,差异均有统计学意义(P<0.05);回归预测模型及ROC分析结果显示:血清Tau蛋白、H-FABP、sST2三指标单独及联合应用预测ACMP患者预后的ROC-AUC(0.95CI)分别为0.734(0.516~0.934)、0.731(0.489~0.957)、0.736(0.497~0.974)、0.885(0.793~0.952),其中联合应用诊断效能最高。结论血清Tau蛋白、H-FABP、sST2水平与ACMP患者的病情严重程度有关,同时也是ACMP患者预后的影响因素,联合检测可提高对ACMP患者预后的预测效能。

Regulating Effects of Different Moxibustion Therapies on Bax and Bcl-2 in Rats with Ulcerative Colitis

Objective： To investigate the improving effects of different moxibustion therapies on colonic injury in rats with ulcerative colitis （UC）, and on the expression of Bcl-2 and Bax mRNA.Methods： Rats model of UC was made with immune methods and local stimulation. The models were treated with different moxibustion therapies 14 times, and then the expression of Bcl-2 and BaxmRNA were tested in the colonic tissue with fluorescent quantitative PCR. Results： （1）General scores of colon and histologic scores of injury were improved in all treatment groups except the model group（P〈0.05, P〈0.01）; （2） The expression of Bcl-2 mRNA in the model rats of UC was higher than that in the normal rats （P〈0.01）, whereas Bax lower （P〈0.01）; （3） After treatments, the expression of Bcl-2mRNA was lower in the herb cake-partitioned moxibustion, ginger-partitioned moxibustion, and garlic-partitioned moxibustion groups than that in the model group （P〈0.05, P〈0.01）, and Bax mRNAin the herb cake-partitioned and ginger-partitioned groups was higher （P〈0.05）. Conclusion： Herb cake-partitioned moxibustion, ginger-partitioned moxibustion, garlic-partitioned moxibustion, andmild moxibustion could improve the pathological injury in UC rats. Herb cake-partitioned moxibustion, ginger-partitioned moxibustion, and garlic-partitioned moxibustion could inhibit theexpression of Bcl-2 mRNA and Bax mRNA in the colonic tissue of UC rats to regulate the ratio of Bcl-2/Bax mRNA.

Bcl-2基因在一氧化碳暴露大鼠脑中的表达

目的 观察一氧化碳 (CO)暴露后大鼠脑中 Bcl- 2基因的分布。方法 用 Wistar大鼠制成CO暴露动物模型 ,随机分组 ,分别在染毒后第 3天、第 2天和当天处死 ,取脑制成切片 ,用地高辛 Bcl- 2探针原位杂交 ,观察 Bcl- 2的信号分布。结果 Bcl- 2 m RNA的表达在海马、大脑皮层和胼胝体比较明显 ,其中海马的表达信号最强 ,大脑皮层次之 ,胼胝体最弱。其他部位呈散在的弱信号。相应区域在各个时间点无明显的差异。对照组未见表达。结论 CO暴露后 ,Bcl- 2在海马、大脑皮层和胼胝体的表达较为明显 ;Bcl- 2在上述部位的表达可能部分的解释 CO中毒迟发性神经损伤的发生。

HO/CO在糖尿病肾病大鼠肾脏组织中的表达及其意义

目的:观察糖尿病肾病模型大鼠血浆一氧化碳(CO)的变化和血红素氧合酶-1(HO-1)、血红素氧合酶-2(HO-2)在肾脏组织的表达,探讨血红素氧合酶/一氧化碳(HO/CO)系统在糖尿病肾病发生和发展中的作用。方法:将24只SD大鼠随机分为对照组和模型组,按Anderson等方法建立糖尿病肾病模型。8周后收集尿液测定24 h的尿蛋白(Upro)和尿肌酐(Ucr);大鼠断头取血测定CO、尿素氮(BUN)和肌酐(Scr),计算肌酐清除值(Ccr);剖取肾脏组织,采用免疫组织化学法检测肾脏组织HO-1和HO-2的表达。结果:与对照组比较,模型组大鼠Upro、BUN和Scr明显升高(P<0.01),Ccr降低(P<0.01);模型组大鼠血浆CO与对照组比较明显降低(P<0.01),肾脏组织HO-1和HO-2的表达强度显著减弱(P<0.01)。结论:HO/CO的调节失常参与了糖尿病肾病的发生和发展。

基因作为一氧化碳中毒诊断标志物的初步研究

目的探讨急性一氧化碳(CO)中毒后大鼠海马组织基因表达的变化,为发现更稳定、可靠的CO中毒诊断标志物提供线索。方法选取雄性Wistar大鼠20只,均为4月龄。随机分为正常对照组(NC组)10只、急性CO中毒组(ACO-P组)10只。用吸入CO气体法建立急性CO中毒模型,用Biostar R40s基因表达谱芯片检测急性CO中毒14 d后大鼠海马组织基因表达变化的情况。结果急性CO中毒大鼠建模成功,ACO-P组大鼠血液中碳氧血红蛋白(COHb)浓度维持在高水平(>50%);NC组大鼠血中COHb浓度在正常水平(<2%)。急性CO中毒14 d后,大鼠海马组织中共有33条基因出现明显表达变化,其中18条基因上调,15条基因下调;这些基因涉及免疫应答、细胞信号转导、传递蛋白、细胞代谢、衰老等多个方面。结论急性CO中毒后14 d发现多条基因出现差异表达,初步为急性CO中毒的诊断提供了较COHb更为稳定、可靠的生物学标志物。

NLR联合sST2对中重度急性CO中毒心肌损伤患者发生院内心血管不良事件的预测价值——一项单中心前瞻性观察性研究

目的探讨中性粒细胞/淋巴细胞比值(NLR)联合可溶性生长刺激表达基因2蛋白(sST2)对中重度急性一氧化碳中毒(ACOP)心肌损伤患者发生院内主要心血管不良事件(MACE)的预测价值。方法采用单中心前瞻性观察性研究方法,选择2016年11月至2020年2月河北医科大学哈励逊国际和平医院急救医学部收治的中重度ACOP合并心肌损伤患者作为研究对象,收集患者的基线资料、入院时NLR、sST2和入院3 d sST2以及其他心肌损伤相关生化指标。根据是否发生MACE将患者分为MACE组和非MACE组,比较两组患者临床资料的差异;各项指标的相关性采用Pearson相关性分析法;采用二元Logistic回归分析影响中重度ACOP心肌损伤患者发生院内MACE的独立危险因素;绘制受试者工作特征曲线(ROC曲线),计算ROC曲线下面积(AUC),分析NLR、sST2单独以及二者联合检测对中重度ACOP心肌损伤患者发生院内MACE的预测价值。结果278例中重度ACOP心肌损伤患者纳入最终分析,MACE发生率11.51%(32/278)。MACE组心肌肌钙蛋白I(cTnI)、血乳酸(Lac)、NLR、入院3 d sST2均明显高于非MACE组〔cTnI(μg/L):0.83±0.15比0.46±0.37,Lac(mmol/L):2.96±1.14比2.43±1.35,NLR:13.14±4.37比9.49±4.21,入院3 d sST2(μg/L):59.88±23.42比39.83±12.60,均P<0.01〕,MACE组与非MACE组入院时sST2比较差异无统计学意义(μg/L:269.09±90.89比240.14±113.02,P>0.05)。Pearson相关性分析显示,NLR与急性生理学与慢性健康状况评分Ⅱ(APACHEⅡ)、入院3 d sST2与APACHEⅡ评分、NLR与入院3 d sST2均呈显著正相关(r值分别为0.226、0.209、0.193,均P<0.01)。二元Logistic回归分析显示,入院3 d sST2和NLR均是影响中重度ACOP心肌损伤患者发生院内MACE的独立危险因素〔优势比(OR)和95%可信区间(95%CI)分别为1.064(1.039~1.090)、1.176(1.066~1.298),均P<0.01〕。ROC曲线分析显示,NLR联合入院3 d sST2对ACOP心肌损伤患者发生院内MACE的预测效能(AUC=0.876)优于NLR(AUC=0.754)和入院3 d sST2(AUC=0.813)。当NLR的最佳临界值为10.02,入院3 d sST2的最佳临界值为43.50μg/L时,预测中重度ACOP心肌损伤患者发生MACE的敏感度为69.8%和86.2%,特异度为74.3%和70.4%,而二者联合检测的特异度和敏感度为83.4%、79.8%。结论NLR和入院3 d sST2是中重度ACOP心肌损伤患者发生院内MACE的独立预测因素,二者联合检测的预测价值好;对于NLR>10.02、入院3 d sST2>43.50μg/L的中重度ACOP心肌损伤患者需警惕发生院内MACE。

外源性一氧化碳释放分子2对大肠杆菌活力及毒力的作用

目的 探讨外源性一氧化碳释放分子2（CORM-2）对大肠杆菌ATCC 25922菌株活力及毒力的作用及可能机制。方法 （1）体外实验1。将菌株按照随机数字表法分为细菌组、细菌＋1.2 mmol/L CORM-2组、细菌＋1.6 mmol/L CORM-2组、细菌＋1.2 mmol/L无活性CORM-2（iCORM-2）组、细菌＋1.6 mmol/L iCORM-2组,每组样本数为6。细菌组不添加任何物质，其余4组添加相应浓度的CORM-2或iCORM-2。于培养0、3、5、8、10、12、16、20、24、27、30、48 h，测定各组菌液的增殖活性，结果以吸光度值（波长为600 nm）表示；同时进行菌落计数。（2）体外实验2。另取菌株，将其分为细菌组和细菌＋0.8 mmol/L CORM-2组，采用基因芯片筛查出大肠杆菌的4个相关基因fliA、dnaK、marA和waaQ进行实时定量PCR（qRT-PCR），检测各基因表达量。（3）在体研究。另取菌株同体外实验1分组及处理，培养至细菌组菌液的吸光度值（波长600 nm）为0.4时，各组收集0.5 mL菌液。将72只C57BL/6小鼠按照随机数字表法分为空白对照组、细菌组、细菌＋1.2 mmol/L CORM-2组、细菌＋1.6 mmol/L CORM-2组、细菌＋1.2 mmol/L iCORM-2组、细菌＋1.6 mmol/L iCORM-2组，每组12只。空白对照组小鼠不行任何处理，其余5组小鼠取对应有或无添加物的0.5 mL菌液进行腹腔注射。注射后对后5组小鼠进行大体观察。注射后6、12 h检测后5组小鼠血清中TNF-α、IL-6水平，注射后12 h收集小鼠肝、肺组织标本检测髓过氧化物酶（MPO）活性。空白对照组小鼠行相同检测。对数据进行重复测量设计方差分析、析因设计方差分析、单因素方差分析和t检验。结果 （1）体外实验1。与细菌组和细菌＋1.2 mmol/L iCORM-2组比较，细菌＋1.2 mmol/L CORM-2组多数时相点细菌增殖活性明显受抑，菌落明显数量减少（F值分别为1170.80、217.52，P值均小于0.01）；与细菌组和细菌＋1.6 mmol/L iCORM-2组比较，细菌＋1.6 mmol/L CORM-2组多数时相点细菌增殖活性亦明显受抑，菌落数量亦明显减少（F值分别为7948.34、14 432.85，P值均小于0.01）。（2）体外实验2。qRT-PCR检测结果显示：与细菌组比较，细菌＋0.8 mmol/L CORM-2组fliA基因表达下调，dnaK、marA和waaQ基因表达上调（t值分别为30.28、-165.54、-168.88、-187.28，P值均小于0.01）。（3）在体研究。细菌组和细菌＋1.2 mmol/L iCORM-2组、细菌＋1.6 mmol/L iCORM-2组小鼠出现诸如精神萎靡、呼吸急促等症状，细菌＋1.2 mmol/L CORM-2组、细菌＋1.6 mmol/L CORM-2组小鼠上述症状轻微或不明显。注射后6 h，细菌组、细菌＋1.2 mmol/L iCORM-2组小鼠血清TNF-α、IL-6水平分别为（647.3±3.8）pg/mL、（3.44±0.22）ng/mL以及（639.3±0.8）pg/mL、（2.47±0.32）ng/mL，明显高于细菌＋1.2 mmol/L CORM-2组［（124.6±10.7）pg/mL、（1.03±0.16）ng/mL，t值为15.22~84.03，P值均小于0.01］。与细菌组和细菌＋1.6 mmol/L iCORM-2组比较，注射后6、12 h细菌＋1.6 mmol/L CORM-2组小鼠血清TNF-α、IL-6水平亦均明显降低（t值为19.27~245.34，P值均小于0.01）。注射后12 h，细菌＋1-2 mmol/L CORM-2组小鼠肝、肺组织中MPO活性明显低于细菌组、细菌＋1.2 mmol/L iCORM-2组，细菌＋1.6 mmol/L CORM-2组小鼠肝、肺组织中MPO活性亦明显低于细菌组、细菌＋1.6 mmol/L iCORM-2组（t值分别为17.36~18.92、2.35~3.61，P值均小于0.01）。结论 CORM-2能够明显抑制大肠杆菌的活力和毒力，其抑制作用可能与其调节大肠杆菌部分重要的靶基因（fliA、dnaK、marA和waaQ）有关。

Investigation of acupuncture in improving sleep,cognitive and emotion based on attenuation of oxidative stress in prefrontal cortex in sleep-deprived rats

Objective To explore whether acupuncture can improve sleep disturbance,cognitive impairment and emotional disorders caused by sleep deprivation,and its association with the attenuation of oxidative stress injury in prefrontal cortex.Methods Fifty-two male Sprague-Dawley rats were randomly divided into a control group(n=10),a model group(n=14),a manual acupuncture(MA)group(n=14),and a sham-MA group(n=14).All the groups were established as sleep deprivation models via the modified multiple platform method,except for the control group.Rats in both the MA group and the sham-MA group received corresponding intervention,respectively.After modeling and intervention,the four groups received three behavioral tests,namely sleep monitoring,by comprehensive lab animal monitoring system(CLAMS),Morris water maze(MWM)test and open-field test(OFT),followed by oxygen free radical level test and Western blot(WB)detection for the expression levels of Bax and Bcl-2.Results The MA group derived more sleep time within 24 h than either the model group or the sham-MA group(both P<0.05).On MWM orientation navigation test day 1,there were no significant differences in escape latency among the control,MA and sham-MA groups(P>0.05),and the escape latency was significantly shorter in these three groups than that in the model group(all P<0.05).On test day 4,the escape latency was markedly shorter in the MA group than that in either the model group or the sham-MA group(both P<0.05);meanwhile,the MA group showed significantly better performance compared with these two groups in space probe test(both P<0.05).In OFT,compared with the control group,there was a significant decline in the horizontal movement score in the other three groups(all P<0.05),and the decrease was more significant in the model group and the sham-MA group than that in the MA group(both P<0.05).The superoxide dismutase(SOD)content was markedly higher and the malondialdehyde(MDA)content was markedly lower in the MA group than those in the model group and the sham-MA group(all P<0.05).Compared with the model group and the sham-MA group,the expression of Bax was significantly lower and the expression of Bcl-2 was significantly higher in the MA group(all P<0.05).Conclusion MA therapy can lengthen the sleep time in sleep-deprived rats and improve learning and memory impairments induced by sleep deprivation,and the underlying mechanism may be associated with the enhancement of antioxidant capacity in the prefrontal cortex and the inhibition of hippocampal neuronal apoptosis.

一氧化碳释放分子-2对乳腺肿瘤大鼠肠屏障损伤的影响

目的观察一氧化碳释放分子-2(CORM-2)对乳腺肿瘤大鼠肠屏障损伤的影响。方法选取60只雌性SD大鼠为研究对象,采用随机数字表法分为假手术组、乳腺肿瘤组、CORM-2组、灭活型CORM-2(iCORM-2)组,各15只。CORM-2组和iCORM-2组均在制作乳腺肿瘤模型1 h前腹腔注射5 mg/kg干预药物。制作模型或术后24 h留取回肠组织,蛋白质印迹法(Western blot)检测回肠组织的咬合蛋白(Occludin)、紧密连接蛋白-1(ZO-1)、闭合蛋白(Claudin3)表达,酶联免疫吸附试验(ELISA)法检测血D-乳酸、肿瘤坏死因子-α(TNF-α)、肠型脂肪酸结合蛋白(I-FABP)、二胺氧化酶(DAO)、髓过氧化物酶(MPO)水平,反转录-聚合酶链反应(RT-PCR)检测小肠黏膜Rho激酶-1 mRNA、ROCK-2 mRNA。分析CORM-2对乳腺肿瘤大鼠肠屏障损伤的影响,应用SPSS 22.0统计软件分析。结果乳腺肿瘤组、iCORM-2组大鼠的Occludin(0.07±0.01、0.13±0.02)、ZO-1(0.09±0.03、0.12±0.02)、Claudin3(0.21±0.03、0.24±0.06)蛋白表达均低于假手术组(0.35±0.04、0.49±0.05、0.64±0.07)和CORM-2组](0.33±0.03、0.23±0.04、0.42±0.05),CORM-2组的ZO-1、Claudin3蛋白表达水平低于假手术组,差异有统计学意义(t=2.389、2.186、2.801,P<0.05)。乳腺肿瘤组、iCORM-2组大鼠的血D-乳酸[(138.28±29.37)、(135.78±20.93)μg/L]、TNF-α[(354.32±23.39、314.43±21.17)ng/L]、I-FABP(585.63±119.34、548.37±132.38)水平均高于假手术组和CORM-2组(65.30±23.19、75.29±32.11),(65.48±10.92、103.28±19.92),(230.19±38.27、306.28±19.22),CORM-2组大鼠的血D-乳酸、TNF-α、I-FABP、MPO水平均高于假手术组,差异有统计学意义(t=2.549、2.495、2.858,P<0.05)。4组大鼠的血DAO水平比较差异无统计学意义(t=2.252,P>0.05)。乳腺肿瘤组、CORM-2、iCORM-2组大鼠小肠黏膜Rho激酶-1 mRNA(1.03±0.04、3.27±0.07、2.21±0.05)、ROCK-2 mRNA(0.78±0.08、2.24±0.12、2.17±0.17)均高于假手术组(2.85±0.08、2.36±0.08),CORM-2组大鼠的小肠黏膜Rho激酶-1 mRNA低于乳腺肿瘤组和iCORM-2组,差异有统计学意义(F=7.831、8.427、4.997,P<0.05)。乳腺肿瘤组、CORM-2、iCORM-2组大鼠的小肠黏膜ROCK-2 mRNA比较差异无统计学意义(F=1.605,P>0.05)。结论CORM-2可缓解乳腺肿瘤大鼠的肠屏障损伤,抑制炎性介质释放,增强肠上皮细胞间的紧密联系,增大肠黏膜通透性,保护肠屏障。

依达拉奉对急性一氧化碳中毒大鼠脑组织超微结构及抗氧化因子表达的影响

目的 观察急性一氧化碳（CO）中毒后大鼠脑组织超微结构及血红素加氧酶-1（HO-1）和核转录因子红细胞2相关因子-2（NRF-2）表达的变化,探讨依达拉奉对中毒后脑损伤的神经保护作用.方法 按文献建立急性CO中毒的动物模型,用透视电镜观察CO中毒后大鼠脑组织超微结构变化,TUNEL染色法检测大鼠脑组织细胞凋亡.免疫组化和免疫荧光双标法检测依达拉奉给药前后大鼠脑组织HO-1和NRF-2的表达变化.结果 CO中毒可引起大鼠脑组织超微结构改变,主要表现为：细胞核内染色质凝聚固缩,细胞质溶解,胞浆内细胞器溶解消失.依达拉奉能明显改善CO中毒后大鼠脑组织超微结构损伤.CO中毒可诱发神经细胞凋亡.随着中毒时间的延长,CO中毒组大鼠脑组织凋亡细胞逐渐增多,但皮质区和纹状体区凋亡细胞数比较差异无统计学意义（P＞0.05）.依达拉奉给药后大鼠脑组织凋亡细胞明显减少,A值也相对降低.HO-1和NRF-2蛋白在正常大鼠脑组织中可见少量表达.CO中毒后两者的表达水平逐渐增加,在皮质区的表达略高于纹状体区,依达拉奉给药后这2种蛋白的表达水平发生变化.结论 依达拉奉可改善中毒后脑组织超微结构损伤,通过抑制神经细胞凋亡,上调HO-1/NRF-2表达,进而对急性CO中毒后脑损伤发挥重要的神经保护作用.

两指标联合对急性中重度一氧化碳中毒心肌损伤患者发生院内心血管不良事件的预测价值

目的探讨可溶性生长刺激表达基因2蛋白(soluble growth stimulating expression gene 2 protein,sST2)联合中性粒细胞/淋巴细胞比值(neutrophil/lymphocyte ratio,NLR)对预测急性中重度一氧化碳中毒(acute carbon monoxide poisoning,ACOP)心肌损伤患者发生院内心血管不良事件的临床意义。方法纳入2017年1月至2019年12月于河北医科大学哈励逊国际和平医院急诊病房及EICU收治的ACOP合并心肌损伤的患者,收集患者基线资料,入院时查血常规计算NLR,入院时、入院3 d酶联免疫吸附法(ELISA)检测sST2(T0sST2、T3dsST2)。根据是否发生心血管不良事件,分为事件组和无事件组,二元logistics回归分析院内心血管不良事件危险因素,受试者工作曲线分析sST2、NLR、sST2联合NLR对预测ACOP心肌损伤患者发生院内心血管不良事件的价值。结果 255例ACOP心肌损伤患者纳入最终分析,事件组29例,发生率11.37%。事件组NLR为(13.38±4.33),无事件组NLR为(9.57±4.22),事件组T3dsST2(61.59±22.67)ng/mL,无事件组T3dsST2(40.52±13.14)ng/mL,差异有统计学意义(均P<0.001);事件组T0sST2(265.34±89.95)ng/mL,无事件组T0sST2(242.43±93.09)ng/mL,差异无统计学意义(P=0.333)。二元logistics回归分析显示,NLR(OR=1.270,95%CI:1.125~1.434,P<0.01)、T3dsST2(OR=1.082,95%CI:1.052~1.114,P<0.01)是ACOP心肌损伤患者发生院内心血管不良事件的独立危险因素。T3dsST2的最佳截断值为44.5 ng/mL,NLR的最佳截断值为12.08,联合应用T3dsST2和NLR预测ACOP心肌损伤患者发生院内心血管不良事件的敏感度为79.3%,特异度为82.7%,曲线下面积为0.857,约登指数为0.620。结论 T3dsST2和NLR是ACOP心肌损伤患者发生院内心血管不良事件的独立危险因素,联合应用T3dsST2和NLR对ACOP心肌损伤患者发生院内心血管不良事件具有较好的预测价值。以T3dsST2为44.5 ng/mL、NLR为12.08为界值,可以作为临床预测ACOP心肌损伤患者发生院内心血管不良事件的早期预测指标。

两指标在一氧化碳中毒性心脏病评估中的应用

目的探讨心率校正后的QT离散度(QTcd)及可溶性生长刺激表达基因2蛋白(sST2)与急性一氧化碳中毒性心脏病患者病情严重程度与预后的关系。方法回顾性分析2017年1月至2020年3月入住本院急救医学部的急性一氧化碳中毒性心脏病患者135例,记录入院即刻、3 h、12 h、2 d、3 d血sST2、肌酸激酶同工酶(CK-MB)及肌钙蛋白I(cTnI)浓度;由专人测量并计算患者入院当天、2 d、3 d及出院时QTcd;根据一氧化碳中毒性心脏病严重程度分为轻度心脏病组(58例)、中度心脏病组(45例)、重度心脏病组(32例),按照是否发生重度心脏病分为重度组(32例)和非重度组(103例),在重度心脏病患者中根据患者是否死亡分为死亡组和存活组。两组间比较采用t检验,相关性分析采用Spearman相关法分析,并绘制受试者工作特征曲线(ROC),分析sST2、QTcd对重度急性一氧化碳中毒性心脏病早期的预测价值及相关性。结果135例患者中有32例为重度中毒性心脏病,发生率为23.7%。重度组sST2自入院时开始升高,cTnI自24 h开始升高,CK-MB自2 d开始升高,检测值均高于非重度组及健康对照组,差异有统计学意义(P<0.05)。治疗前各中毒组患者sST2、QTcd与非重度组、健康对照组比较,差异有统计学意义(P<0.05)。中毒后2 d、3 d不同程度中毒组之间各指标比较,差异有统计学意义(P<0.05)。ROC曲线分析显示,sST2曲线下面积为0.726,95%CI为0.555~0.898,敏感度56.3%,特异度94.1%,截断值为88.5 ng/mL;QTcd曲线下面积为0.745,95%CI为0.602~0.889,敏感度56.3%,特异度82.4%,截断值为68.5 ms;两者联合检测曲线下面积为0.939,95%CI为0.874~1.000,敏感度81.3%,特异度91.2%。结论急性一氧化碳中毒性心脏病患者sST2水平升高时间早于cTnI、CK-MB,sST2与QTcd两者联合观测可作为早期预测急性一氧化碳中毒性心脏病及其严重程度的指标。