槐杞黄颗粒辅助治疗儿童支气管哮喘疗效的真实世界研究

目的 观察槐杞黄颗粒辅助治疗儿童支气管哮喘的疗效。 方法 采取多中心、前瞻性、登记注册的真实世界研究方法,于全国21家医院门诊依序纳入已明确诊断支气管哮喘、年龄2~5岁、采用以下两种治疗方案之一的患儿：使用哮喘长期控制药物,即吸入激素和/或白三烯受体拮抗剂,但未使用槐杞黄颗粒（控制治疗组,n=390）;使用哮喘长期控制药物,同时加用槐杞黄颗粒（联合治疗组,n=1 014）。收集所有患儿的个人及临床资料,于治疗后第4、8、12、20、28、36周进行门诊或电话随访。随访内容包括哮喘发作情况、鼻炎症状等,并对随访观察指标的变化进行统计学分析。 结果 入组前两组患儿哮喘发作次数、天数及鼻炎发作天数比较差异均无统计学意义（P>0.05）。经治疗后,联合治疗组患儿每月哮喘发作天数、重度哮喘发作次数及鼻炎发作天数均显著少于控制治疗组（P<0.05）。两组患儿不良反应发生率比较差异无统计学意义（P=0.667）。 结论 在使用哮喘长期控制药物治疗基础上加用槐杞黄颗粒能够改善支气管哮喘患儿的临床症状及伴发的鼻炎症状,提高哮喘控制水平,且在治疗过程中使用安全,无明显不良反应。 引用格式:     

糖皮质激素治疗对重症肌无力患者BAFF和IL-6水平的影响

目的 探讨糖皮质激素(GC)对新发病重症肌无力(MG)患者外周血B淋巴细胞刺激因子(BAFF)、白介素6(IL-6)水平及临床疗效的影响.方法 采用ELISA法检测26例MG患者GC治疗前、后及正常对照组血清BAFF、IL-6水平,并观察GC治疗的临床效果.结果 MG患者外周血清IL-6、BAFF水平明显高于正常对照组水平(P<0.05),激素治疗2个月后,MG患者外周血BAFF、IL-6水平显著下降(t=9.79 or t=3.45,P<0.01),治疗前后IL-6变化与肌无力相对评分存在显著正相关性(r=0.559,P<0.01).结论 MG患者血清IL-6水平可能与病情严重程度呈正相关,GC可通过抑制IL-6、BAFF分泌,有效调节MG患者细胞免疫功能.     

2型糖尿病对急性脑梗死患者脑干听觉诱发电位及预后的影响

目的探讨2型糖尿病对急性脑梗死患者脑干听觉诱发电位(BAEP)及神经功能的影响。方法选择急性期脑梗死患者154例,包括2型糖尿病合并脑梗死患者(DMCI)74例和非糖尿病脑梗死患者(NDMCI)80例,全部患者发病72h内均行BAEP检查和美国国立卫生院卒中量表(NIHSS)评分。结果 DMCI组与NDMCI组患者BAEP异常率分别为81.2%和63.8%,差异有统计学意义(P0.05);DMCI组患者BAEP主波潜伏期和波间期均较NDMCI组患者明显延长(P0.05);2组患者BAEP异常率与NIHSS评分呈正相关。结论 2型糖尿病加重急性期脑梗死患者脑干功能损伤,BAEP可作为急性期脑梗死患者预后的有效评估指标。     

促红细胞生成素及其衍生物在局灶性脑缺血中的神经保护作用

目的 探讨氨甲酰促红细胞生成素(CEPO)对缺血性脑损伤的保护作用及机制,并与促红细胞生成素(EPO)进行比较. 方法 用腔内线栓法制造小鼠脑缺血90min再灌注模型,根据颈动脉注射药物的不同分为对照组(注射生理盐水)、EPO 5 μg/kg组、EPO 50 μg/kg组、CEPO 50μg/kg组,每组6只,用脑血流激光多普勒监测脑缺血过程中脑血流的变化;用甲酚紫染色法显示脑梗死区并用imageJ软件计算梗死体积和脑水肿体积;用TUNEL法观察凋亡细胞;用免疫组织化学方法 观察脑缺血再灌注后诱导型一氧化氮合酶(iNOS)的改变. 结果与对照组比较,EPO 50μg/kg组和CEPO 50μg/kg组的脑梗死体积、脑水肿体积、神经功能缺损评分明显减少,差异均有统计学意义(P＜0.05).EPO 50μg/kg组、CEPO 50 μg/kg组缺血皮层iNOS阳性细胞数与对照组比较,差异均有统计学意义(P＜0.05).EPO 50μg/kg组的凋亡细胞[(43.6±10.1)个]、CEPO 50 μg/kg组的凋亡细胞[(40.5±9.8)个]与对照组[(94.2±15.2)个]比较,差异均有统计学意义(P＜0.05). 结论 低剂量的EPO(5μg/kg)无脑保护作用,CEPO 50 μg/kg与较高剂量EPO(50μg/kg)具有相似的增加脑缺血再灌注后的脑血流、减少神经功能缺损评分、减少梗死体积、缩小脑水肿体积和抗细胞凋亡作用,它们通过减少iNOS的表达而发挥神经保护作用.     

The effects of immediate and delayed mild hypothermia on endogenous antioxidant enzymes and energy metabolites following global cerebral ischemia

Background: The optimal time window for the administration of hypothermia following cerebral ischemia has been studied for decades, with disparity outcomes. In this study, the efficacy of mild brain hypothermia beginning at different time intervals on brain endogenous antioxidant enzyme and energy metabolites was investigated in a model of global cerebral ischemia. Methods: Forty-eight male Sprague-Dawley rats were divided into a sham-operated group, a normothermia (37-38 °C) ischemic group and a mild hypothermic (31-32°C) ischemia groups. Rats in the last group were subdivided into four groups: 240 minutes of hypothermia, 30 minutes of normothermia plus 210 minutes of hypothermia, 60 minutes of normothermia plus 180 minutes of hypothermia, and 90 minutes of normothermia plus 150 minutes of hypothermia (n=8). Global cerebral ischemia was established using the Pulsinelli four- vessel occlusion model for 20 minutes and mild hypothermia was applied after 20 minutes of ischemia. Brain tissue was collected following 20 minutes of cerebral ischemia and 240 minutes of reperfusion, and used to measure the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and adenosine triphosphate (ATP). Results: Mild hypothermia that was started within 0 to 60 minutes delayed the consumption of SOD, GSH-Px, GSH, and ATP (P<0.05 or P<0.01) in ischemic tissue, as compared to a normothermic ischemia group. In contrast, mild hypothermia beginning at 90 minutes had little effect on the levels of SOD, GSH-Px, GSH, and ATP (P>0.05). Conclusions: Postischemic mild brain hypothermia can significantly delay the consumption of endogenous antioxidant enzymes and energy metabolites,that are critical to the process of cerebral protection by mild hypothermia. These results show that mild hypothermia limits ischemic injury if started within 60 minute,but loses its protective effects when delayed until 90 minutes following cerebral ischemia.     

Bcl-2基因修饰的骨髓间充质干细胞移植治疗脑缺血的实验研究

目的观察Bcl-2基因修饰的骨髓间充质干细胞（MSCs）移植对大鼠脑缺血性损伤的治疗作用及对移植的MSCs保护性作用。方法将114只SD大鼠随机分为假手术组、Model组、MSCs组和Bcl-2-MSCs组;线栓法制作大鼠一侧大脑中动脉缺血再灌注模型,MSCs组及Bcl-2-MSCs组在缺血24h后经尾静脉注射方式移植BrdU标记的MSCs及人Bcl-2基因修饰的MSCs;分别于术后1、7、14、28d对各组大鼠进行神经功能缺损评分（NSS）;在脑缺血14d应用TTC法观察梗死灶体积;BrdU和TUNEL免疫荧光双重标记检测移植的MSCs凋亡情况;Westernblot检测大鼠脑梗死周边区Bcl-2蛋白的表达;HE染色观察脑组织病理形态。结果MSCs-Bcl-2组和MSCs组NSS评分、脑梗死体积百分比、TUNEL阳性细胞数较Model组低（P〈0.05）,且MSCs-Bcl-2组比MSCs组更低,BrdU阳性细胞数较多（P〈0.05）;MSCs-Bcl-2组BrdU和TUNEL免疫荧光双标细胞数稍多,但差异不显著（P〉0.05）,而MSCs-Bcl-2组BrdU和TUNEL免疫荧光双标细胞占BrdU阳性细胞百分比明显较低（P〈0.05）。MSCs-Bcl-2组Bcl-2蛋白呈持续较高水平的表达,和MSCs组同时间点相比差异有统计学意义（P〈0.05）。HE染色示MSCs组和MSCs-Bcl-2组脑组织损伤及细胞丢失较轻,MSCs-Bcl-2组更明显,脑梗死周边区均未见到核大、浓染的异形细胞。结论Bcl-2基因修饰的MSCs移植较MSCs移植能进一步改善脑缺血大鼠的神经功能,减少脑梗死体积;其机制为Bcl-2基因修饰使MSCs持续、稳定表达一定量的Bcl-2蛋白,从而能保护移植的MSCs,减少其凋亡,增加其存活。     

Over-expression of VEGF in marrow stromal cells promotes angiogenesis in rats with cerebral infarction via the synergistic effects of VEGF and Ang-2

This study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction.MSCs were isolated by using a direct adherent method and cultured.Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection.The transfection efficiency was measured by the optical density method.The protein expression of VEGF gene before and after transfection was measured by Western blotting.SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion.Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction.ELISAs were used to measure the levels of VEGF in the rat cerebral tissues.The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically oserved.Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting.VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%.ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction,peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days.The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group.Moreover,the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day.The immunohistochemical results showed that 10 days after the infarction,the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group.Moreover,the VEGF level was higher in the VEGF-MSC group th     

Involvement of PI3K/Akt pathway in the neuroprotective effect of sonic hedgehog on cortical neurons under oxidative stress

The Sonic hedgehog (SHH) signaling pathway plays a pivotal role in neurogenesis and brain damage repair. Our previous work demonstrated that the SHH signaling pathway was involved in the neuroprotection of cortical neurons against oxidative stress. The present study was aimed to further examine the underlying mechanism. The cortical neurons were obtained from one-day old Sprague-Dawley neonate rats. Hydrogen peroxide (H2O2, 100 μmol/L) was used to treat neurons for 24h to induce oxidative stress. Exogenous SHH (3μg/mL) was employed to activate the SHH pathway, and cyclopamine (20 μmol/L), a specific SHH signal inhibitor, to block SHH pathway. LY294002 (20 μmol/L) were used to pretreat the neurons 30 min before H2O2 treatment and selectively inhibit the phosphatidylinositol 3-kinase (PI3K)/Akt pathway. The cell viability was measured by MTT and apoptosis rate by flow cytometry analysis. The expression of p38, p-p38, ERK, p-ERK, Akt, p-Akt, Bcl-2, and Bax in neurons was detected by immunoblotting. The results showed that as compared with H2O2 treatment, exogenous SHH could increase the expression of p-Akt by 20% and decrease the expression of p-ERK by 33%. SHH exerted no significant effect on p38 mitogen-activated protein kinase (p38 MAPK) pathway. Blockade of PI3K/Akt pathway by LY294002 decreased the cell viability by 17% and increased the cell apoptosis rate by 2-fold. LY294002 treatment could up-regulate the expression of the proapoptotic gene Bax by 12% and down-regulate the expression of the antiapoptotic gene Bcl-2 by 54%. In conclusion, SHH pathway may activate PI3K/Akt pathway and inhibit the activation of the ERK pathway in neurons under oxidative stress. The PI3K/Akt pathway plays a key role in the neuroprotection of SHH. SHH/PI3K/Bcl-2 pathway may be implicated in the protection of neurons against H2O2-induced apoptosis.     

神经调节素对大鼠脑缺血损伤的预防性治疗作用和机制

目的探讨神经调节素（NRG-1 β）对大鼠脑缺血损伤的预防性治疗作用和机制。方法Wistar大鼠150只，应用线栓法建立大鼠大脑中动脉闭塞（MCAO）模型，随机分为对照组和治疗组，在MCAO前经颈内动脉注射NRG-1 β（2μg/kg）进行预防性治疗。在MCAO后0h、0．5h、1．0h、1．5h和2．0h分别用干湿重法测定脑组织含水量，氯化三苯基四氮唑（TYC）染色测定脑梗塞体积，细胞凋亡用末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记（TUNEL）法，早期生长反应基因-1（Egr-1）表达水平用免疫组化检测。结果大脑中动脉阻断后，动物脑组织含水量和梗塞面积随着缺血时间的延长而逐渐增加，NRG预处理组脑组织含水量低于对照组，梗塞范围也明显缩小。脑缺血可诱导脑组织细胞凋亡和Egr-1表达，随着缺血时间的延长，凋亡细胞和Egr-1阳性细胞数明显增多；NRG预处理能明显减少缺血诱导的细胞凋亡数，增加Egr-1的表达水平。结论NRG-1 β可能通过抑制凋亡途径和诱导Egr-1表达水平，对缺血性脑损伤具有积极的保护作用。     

血清MMP-9与脑梗死患者及其颈动脉粥样硬化斑块稳定性的关系研究

目的检测具有不同性质颈动脉粥样硬化斑块的脑梗死患者的血清基质金属蛋白酶-9（MMP-9）水平,探讨颈动脉粥样硬化斑块稳定性及相关炎性标志物MMP-9水平与脑梗死的关系。方法采用彩色多普勒超声检查48例颈内动脉系统的急性脑梗死患者（CI组）颈动脉粥样硬化斑块,同时检测患者血清MMP-9水平,并与20例慢性脑供血不足患者（CCCI组）及20例体检健康者（对照组）比较;根据斑块性质将CI组分为不稳定斑块组、稳定斑块组及无斑块组3个亚组,并进行组内比较。结果脑梗死组斑块检出率、不稳定斑块率及血清MMP9水平均明显高于慢性脑供血不足组及对照组（P〈0．05）;脑梗死不稳定斑块组MMP-9水平显著高于脑梗死稳定斑块组,脑梗死稳定斑块组高于脑梗死无斑块组（P〈0．01）。结论颈动脉粥样硬化斑块及其稳定性与脑梗死发生有密切关系,具有不同性质颈动脉斑块的脑梗死患者的血清MMP-9水平存在差异,MMP-9可能是不稳定性粥样硬化斑块及脑梗死的一个潜在的血清标志物。