卡维地洛对免疫性心肌炎小鼠心肌凋亡调控蛋白影响

目的 观察卡维地洛对免疫性心肌炎小鼠心肌凋亡蛋白Bcl-2、Bax及Fas的影响,探讨其在免疫性心肌炎中的作用.方法 60只4～5周龄近交系雄性BALB/C小鼠,随机将小鼠分为3组:免疫性心肌炎模型组(模型组),卡维地洛干预组(干预组),空白对照组(对照组),每组20只.眼球取血后处死,留取心脏及血液标本.光镜观察各组小鼠心肌组织病理学改变,电镜观察心肌细胞超微结构,化学发光免疫法检测血清心肌肌钙蛋白I(cTnI)水平;免疫组化法检测心肌Bcl-2、Bax及Fas蛋白表达的含量;原位缺口末端标记(TUNEL)法检测心肌细胞凋亡情况.结果 模型组小鼠心肌细胞光镜下见大量炎症细胞浸润,电镜下可见细胞核固缩、染色质边集,对照组小鼠心肌细胞光镜下未见炎症细胞浸润,电镜下染色质边集不明显.模型组Bcl-2、Bax及Fas蛋白表达较对照组高(23.48±2.24 vs 6.64±1.60,26.15±2.02 vs 5.09±0.85,21.22±3.62 vs 5.86±1.37,P＜0.01);干预组与模型组比较,小鼠心肌组织炎症积分轻(2.60±0.3l vs 2.02±0.26,P＜0.05),电镜下细胞核固缩轻,心肌细胞Bcl-2、Bax及Fas蛋白表达低(17.13±1.94 vs 23.48±2.24,17.66±2.62 vs26.15±2.02,16.79±2.83 vs 21.22±3.62,P＜0.05),凋亡指数低[(16.61±4.67)%vs(24.51±4.70)%,P＜0.05],cTn-I水平低[(1.878±0.48)ng/ml vs(1.102±0.23)ng/ml,P＜0.05].结论 卡维地洛可减轻免疫性心肌炎小鼠心肌细胞凋亡,对免疫性心肌炎起一定的保护作用.

Abstract：

Objective To observe the effects of carvedilol on the expression of Bcl-2, Bax and Fas in autoimmune myocarditis (AM).Methods A total of 60 inbred male BALB/C mice 4-5 weeks of age were divided at random into 3 groups as follows: AM group ( n = 20), carvedilol group ( n = 20 ) and control group (n = 20).The mice were sacrificed after gathering blood specimens by taking out the eyeballs and hearts tissue.The histological and ultrastructural changes were observed under light microscope and electron microscope.The concentrations of cardiac troponin Ⅰ (cTn Ⅰ ) were detected by chemiluminescence immunoassay (CLIA).Immunohistochemistry (IHC) was performed to analyze the contents of Bcl-2, Bax and Fas, TUNEL to detect the apoptotic index in myocardial cells.Results There were large number of lymphocyte and monocyte infiltrates under light microscope and karyopyknosis and chromatin gathered along the nuclear membrane under electron microscope in AM group.There were no inflammations and chromatin gathering in group C.Compared with control group, the Bcl-2, Bax and Fas protein expression significantly elevated in AM group ( 23.48 ± 2.24 vs.6.64 ± 1.60,26.15 ± 2.02 vs.5.09 ± 0.85,21.22 ± 3.62 vs.5.86 ± 1.37, P ＜ 0.0l ) . The histopathologic scores ( 2.60 ± 0.31 vs.2.02 ± 0.26, P ＜ 0.05 ) and karyopyknosis of carvedilol group decreased as compared with AM group.The Bcl-2, Bax and Fas protein expression (17.13 ±1.94 vs.23.48 ±2.24,17.66 ±2.62 vs.26.15 ±2.02,16.79 ±2.83 vs.21.22 ±3.62, P ＜ 0.05 ), AI [( 16.61 ± 4.67 ) % vs.( 24.51 ± 4.70) %, P ＜ 0.05]and contents of cTnI [( 1.878± 0.48 ) ng/ml vs. ( 1.102 ± 0.23 ) ng/ml, P ＜ 0.05]also decreased in carvedilol group compared with AM group.Conclusion Carvedilol could protect against AM by alleviating cardiomyocyte apoptosis.     

吸入高浓度氧对大鼠肺泡发育过程促红细胞生成素受体的影响

目的 探讨大鼠肺泡发育阶段促红细胞生成素受体(erythropoietin receptor,EPOR)的动态变化以及吸入高浓度氧对EPOR的影响.方法 将48只新生Wistar大鼠生后12 h内随机分为空气组和高氧组.高氧组将动物置于自制密闭氧箱中,持续输入氧气,FiO2=0.85±0.02.在实验3,7和14 d每组随机选取8只处死,采集标本.采用HE染色观察肺组织病理改变,放射状肺泡计数评价肺泡化程度,免疫组织化学法检测肺组织血小板内皮细胞黏附分子-1(platelet endothelial cell adhesion molecule-1,PECAM-1)及EPOR表达,PT-PCR及Western blot法检测肺组织EPOR基因和蛋白表达.结果 高氧组3 d时肺毛细血管扩张、充血,7 d时肺泡体积增大、数量减少,14 d时肺泡数量及毛细血管进一步减少.空气组RAC值随日龄增长而增加,与空气组相比,高氧组7 d时RAC值降低[(6.85±104):(7.33±1.0),P＜0.01],差异在14 d更显著[(6.20±1.58):(9.07±0.69),P＜0.001].空气组PECAM-1表达随日龄增长逐渐增强,高氧组7 d和14 d PECAM-1表达较空气组明显减弱[(15.14±1.51):(31.47±2.43),(11.04±1.76):(41.41±3.83),P＜0.001].空气组EPOR染色在生后3 d最强,随日龄增长逐渐减弱,与空气组相比,高氧组各时点EPOR表达均明显减弱[(1.62±0.04):(1.82±0.06),P＜0.05;(0.48±0.01):(1.10±0.07),(0.39±0.04):(0.87±0.03),P＜0.001].空气组EPOR mRNA表达在生后3 d最强,高氧组各时点EPOR mRNA表达较空气组明显减弱[(0.87±0.07):(1.1±0.17),(0.18±0.07):(0.36±0.08),P＜0.01;(0.14±0.05):(0.36±0.09),P＜0.001].结论 EPOR可能在正常肺泡发育过程发挥调节作用,吸入高浓度氧导致的肺泡及血管发育阻滞可能与EPOR及PECAM-1蛋白表达减弱有关.

Abstract：

Objective Oxygen toxicity is thought to be a major contributing factor in the pathogenesis ofbronchopulmonary dysplasia (BPD). Animal experiments reveal that erythropoietin (EPO) may have protective effects against hyperoxic lung injury, but the mechanisms remain unknown.The aim of this study was to evaluate effects of hyperoxia on erythropoietin receptor expression in lung development of neonatal rats.Methods Several litters of Wistar pups were pooled together within 12 hours after birth and randomly divided into two groups (n = 24 in each ): air-exposed control group and hyperoxia-exposed group. In hyperoxia-exposed group, the rats were exposed to 85% oxygen. Pups ( n = 8 ) from each group were sacrificed on postnatal days 3, 7, and 14.The pulmonary histologcal and morphometric changes were observed after hematoxylin-eosin (HE) staining under light microscope. Radical alveolar counts ( RAC )were compared between the two groups to evaluate the differences of aveolarization. Expressions of platelet endothelial cell adhesion molecule-1 ( PECAM-1 ) and erythropoietin receptor (EPOR) in lung tissue were measured by immunohistochemistry.Expressions of EPOR mRNA and EPOR protein were measured by RTPCR and Western blotting. Results In hyperoxia-exposed group, there were a few inflammatory cells infiltration in interstitium on day 3 and inflammatory response worsened on day 7. Alveolar and capillary hypoplasia and interstitial fibrosis were evident on day 14.RAC increased in air-exposed control group along with the age in days. RAC decreased from day 7 in hyperoxia-exposed group compared with air-exposed control group [( 6.85 ± 1.04 ) vs.( 7.33 ± 1.0 ), P ＜ 0.01], which was more evident on day 14 [( 6.20 ±1.58 ) vs.(9.07 ± 0.69 ), P ＜ 0.001].Expression of PECAM-1 protein increased in air-exposed control group along with the age in days.But in hyperoxia-exposed group, it decreased on day 7 and 14 [( 15.14 ±1.51) vs.(31.47 ±2.43),(11.04 ± 1.76)vs.(41.41 ±3.83),P＜0.001]compared with air-exposed control group.Expression of EPOR on day 3 in air-exposed control group was the strongest and weakened gradually with the increase of postnatal days.Expression of EPOR in hyperoxia-exposed group decreased on day 3 and became more evident on day 7 and day 14 compared with air-exposed control group [( 1.62 ±0.04) vs.(1.82±0.06), P＜0.05;(0.48 ±0.01)vs.(1.10±0.07), (0.39±0.04) vs.(0.87±0.03 ) ,P ＜0.001].Expression of EPOR mRNA on day 3 in air-exposed control group was the strongest and was decreased significantly in hyperoxia-exposed group compared with air-exposed control group at all time points [(0.87±0.07)vs.(1.1±0.17),(0.18±0.07)vs.(0.36±0.08),P＜0.01;(0.14±0.05)vs.(0.36 ± 0.09 ), P ＜ 0.001]. Conclusions EPOR may participate in the modulation of normal lung development.Depressed expression of EPOR and PECAM-1 may be involved in the pathogenesis of alveolar and capillary hypoplasia induced by hyperoxia.     

过敏性紫癜患儿血清IgA1对人脐血管内皮细胞凋亡影响的实验研究

目的 探讨过敏性紫癜(HSP)患儿血清IgA1对脐静脉内皮细胞(HUVEC)凋亡的影响.方法 培养人HUVEC,分为HSP组、正常对照组和空白对照组.亲和层析法分离HSP患儿及正常儿童血清IgA1,分别用50、100、200 μg/ml予以刺激HUVEC,空白对照组采用RPMI-1640培养.6、12、24 h后分别行流式细胞仪和TUNEL法检测细胞凋亡率.Real time PCR和Western blot检测HUVEC中Fas和Caspase-3表达情况.结果 HSP患儿的IgA1(200 μg/ml)可诱导脐静脉内皮细胞发生凋亡,其凋亡率显著高于空白对照组[(14.77 ±2.23)％ vs.(2.25 ±0.77)％,P＜0.01];健康对照的IgA1(200μg/ml)也可诱导脐静脉内皮细胞发生凋亡,凋亡率也显著高于空白对照组[(7.97±1.48)％ vs.(2.25±0.77)％,P＜0.01].HSP患儿的IgA1诱导HUVEC的凋亡呈浓度依赖及时间依赖性.而且HSP患儿的IgA1能显著促进HUVEC中Caspase-3及Fas的表达(P＜0.01).结论 HSP患儿体内IgA1可诱导HUVEC凋亡,其可能参与了HSP发生发展的过程.     

Salubrinal抑制脂多糖诱导H9C2心肌细胞凋亡的保护机制

目的 研究脓毒症中内质网应激诱导心肌细胞凋亡的机制.方法 建立脂多糖(lipopolysacchride,LPS)H9C2心肌细胞脓毒症模型.通过实时定量聚合酶链反应(RTq-PCR)和流式细胞技术分别观察LPS干预H9C2心肌细胞24 h后的促凋亡蛋白CHOP mRNA、线粒体膜电位(mitochondrial membrane potential,MMP)和细胞凋亡的变化.使用内质网应激抑制剂Salubrinal预处理H9C2心肌细胞,观察以上指标变化.结果 LPS组CHOP mRNA表达(2.40±0.30)与对照组比较(1.00±0)显著上调(P＜0.05),LPS组MMP(1.07 ±0.33)与对照组(2.62±0.51)比较显著下降(P＜0.05),LPS组细胞凋亡率[(16.58±2.71)％]与对照组[(3.85±1.07)％]比较显著上升(P＜0.05).Salubrinal+ LPS组CHOP mRNA表达(1.60±0.23)与LPS组(2.40±0.30)比较显著下调(P＜0.05),Salubrinal+ LPS组MMP(1.85 ±0.31)与LPS组比较,显著上调(P＜0.05);Salubrinal+ LPS组细胞凋亡率[(6.05±1.48)％]与LPS组比较,显著下调(P＜0.05).结论 在H9C2心肌细胞脓毒症模型中,Salubrinal通过抑制LPS诱导CHOP mRNA的表达,减轻线粒体的损伤,从而降低心肌细胞凋亡率.脓毒症心肌细胞中内质网应激可能通过线粒体途径诱导细胞凋亡.     

白藜芦醇对大鼠肠缺血再灌注致肠黏膜通透性改变的影响

目的 探讨白藜芦醇(RES)对肠缺血再灌注致肠黏膜损伤大鼠的保护作用及其可能机制.方法 成年雄性SD大鼠24只随机分为假手术组、缺血再灌注损伤组、RES治疗组.假手术组仅分离肠系膜上动脉(SMA)而根部不夹闭.肠缺血再灌注损伤组和RES治疗组均用无损伤血管夹夹闭SMA根部,分别立即经阴茎背静脉注射9g·L-1盐水、RES( 20 mg·kg-1),45 min后放松血管夹形成再灌注.各组大鼠均于制模后6h采集静脉血和回肠标本.检测血清二胺氧化酶(DAO)及小肠脂肪酸结合蛋白(IFABP)水平,应用原位末端转移酶标记法检测肠黏膜上皮细胞凋亡率,HE染色法观察肠组织病理损伤情况.结果 肠缺血再灌注6h后,RES治疗组DAO及IFABP水平与肠缺血再灌注损伤组比较显著减少[DAO:(1 650±1 150)U·L-1vs(2920±1 520)U·L-1;IFABP:(845.12±123.86) μg·L-1vs(1 443.76±174.62) μg·L-1,Pa＜0.05],但二组较假手术组[(630±150)U·L-1,(26.76±4.86)μg·L-1)]均显著增加(Pa＜0.05).假手术组大鼠肠绒毛顶部、固有层和黏膜下层只有少量散在分布的凋亡细胞,缺血再灌注损伤组大鼠肠黏膜凋亡阳性细胞数量较假手术组明显增加[(66.63±1.71)％ vs (9.60±1.76)％,P＜0.05],分布范围从绒毛顶部扩大到中、底部,固有层和黏膜下层,细胞凋亡亦明显加重;RES组大鼠肠黏膜细胞凋亡率[(46.72±1.50)％]明显低于缺血再灌注损伤组(P＜0.05).结论 RES对肠缺血再灌注损伤具有保护作用,其机制可能与其抑制肠黏膜上皮细胞凋亡有关.     

锌指蛋白A20在炎症性肠病患儿肠黏膜中的表达及意义

目的 探讨炎症性肠病(IBD)患儿肠道炎症反应与锌指蛋白A20(A20)表达水平之间的关系.方法 收集2008至2010年就诊于我院并行肠镜检查的患儿肠道黏膜标本共57份.将标本分为正常对照组(n=16)、IBD缓解期组(n=12)、IBD活动期组(n=13)和非IBD肠炎组(n=16).内镜下取各组患儿末端回肠黏膜标本,采用荧光定量PCR和免疫组化法检测A20、NF-κB、IL-6、IL-8的表达水平.结果 (1)NF-κB、A20在正常对照组肠黏膜中仅微量表达,IBD活动期组和非IBD肠炎组NF-κB、A20表达水平明显高于正常对照组(P均＜0.01);(2)IBD缓解期组较正常对照组NF-κB[(9.35±4.84)%vs(0.57±0.44)%,P＜0.01]、IL-6(t'=1.34,P＞0.05)、IL-8(t=1.38,P＞0.05)表达水平高,而A20在mRNA水平(t=1.03,P＞0.05)和蛋白水平[(0.36±0.18)%vs(0.87±0.29)%,P＜0.01]上表达均偏低;(3)与非IBD肠炎组相比,IBD活动期组NF-κB[(24.17±11.27)%vs(55.29±21.84)%,P＜0.01]、IL-6(t=2.22,P＜0.05)、IL-8(t=2.97,P＜0.01)表达水平明显升高,而A20在mRNA(t=2.26,P＜0.05)和蛋白水平[(29.23±11.70)%vs(16.8l±5.90)%,P＜0.01]上表达均较低;(4)IBD缓解期组与非IBD肠炎组相比,IL-6、IL-8表达水平差异无统计学意义(t'值和t值分别为0.03和0.28,P均＞0.05),而A20在mRNA水平(t=4.42,P＜0.01)和蛋白水平[(29.23±11.70)%vs(0.47±0.25)%,P＜0.01]上表达均较低.结论 IBD患儿存在肠道炎症反应过度而A20表达水平上调不足的现象;A20表达水平的异常可能参与了IBD的发生和发展.

Abstract：

Objective It is demonstrated that excessive activation of NF-κB is central to the pathogenesis of inflammatory bowel disease(IBD).Zinc finger protein A20(A20)is a key player in the negative feedback regulation of NF-κB signaling in response to multiple stimuli and has been described as central gatekeeper in inflammation and immunity.Mice genetically deficient in A20 develop severe intestinal inflammation and have increased susceptibility to dextran sodium sulfate(DSS)-induced colitis.Few studies have been done to explore the role of A20 in the pathogenesis of IBD.To clarify the relationship between intestinal inflammation and the expression level of A20 in IBD patients,the expression level of A20 and a series of inflammatory cytokines,such as NF-κB,IL-6,and IL-8,in children with IBD and controls were examined.Method Terminal ileal mucosal samples were obtained via endoscopy. Fifty-seven mucosal samples were divided into 4 groups:normal control group(n = 16),IBD remission group(n = 12),IBD active group(n = 13)and non-IBD enteritis group(n = 16).According to disease activity index scores,the IBD patients were divided into IBD remission group and IBD active group. Normal control group was consisted of patients with functional bowel disorders or intestinal polyps.Non-IBD enteritis was defined as changes in which endoscopy and histological examination showed inflammatory changes but could not be diagnosed as IBD.Real-time PCR was adopted for detecting the mRNA levels of A20,IL-6 and IL-8.Meanwhile immunohistochemistry was performed to measure the expression of A20 and NF-κB.Result (1)The expression of A20 and NF-κB were very low in normal control group,but significantly up-regulated in IBD active group and non-IBD enteritis group(P＜ 0.01 for beth);(2)Compared with normal control group,expression of NF-κB [(9.35±4.84)% vs.(0.57±0.44)%,P＜0.01],IL-6(t' = 1.34,P ＞0.05),IL-8(t = 1.38,P ＞0.05)increased in IBD remission group,while the expression of A20 in both mRNA(t = 1.03,P ＞ 0.05)and protein levels [(0.36±0.18)% vs.(0.87±0.29)%,P＜ 0.01]decreased;(3)Compared with non-IBD enteritis group,although the expression of NF-κB [(24.17±11.27)% vs.(55.29±21.84)%,P＜0.01],IL-6(t =2.22,P＜0.05),IL-8(t=2.97,P＜0.01)were highly increased in IBD active group,the expression of A20 in both mRNA(t =2.26,P＜0.05)and protein levels [(29.23±11.70)% vs.(16.81±5.90)%,P＜ 0.01] significantly decreased;(4)The expression of IL-6,IL-8 were similar in IBD remission group and non-IBD enteritis group(both P ＞0.05),but the expression of A20 was much lower in both mRNA(t =4.42,P＜0.01)and protein levels [(29.23±11.70)% vs.(0.47±0.25)%,P＜ 0.01] in IBD remission group.Conclusion The results demonstrate that there is an excessive inflammatory response but insufficient up-regulation of A20 expression in IBD patients.Low levels expression of A20 may play an important role in the pathogenesis of IBD.     

核因子-kB信号途径活化对川崎病冠状动脉损害作用的研究

目的 探讨核因子-kB(NF-kB)信号途径在川崎病(KD)冠状动脉免疫损害中的作用.方法 2004年10月至2005年12月武汉市儿童医院收治的KD患儿48例,将患儿分为两组:冠状动脉损害亚组(CALs亚组)11例,无冠状动脉损害亚组(Non-CALs亚组)37例.感染对照组:同年龄组感染发热住院患儿18例.健康对照组:同年龄组健康体检儿童12例.采用免疫印迹(Western blot)法检测外周血单个核细胞(PBMC)中NF-KBp65及IkBα蛋白表达;采用逆转录聚合酶链反应(RT-PCR)检测TNF-α、MCP-lmRNA表达.结果 (1)KD组NF-kBp65明显高于两对照组[(16.53±4.81 vs.(8.37±3.15,0.33±0.05)(P<0.001)].CALs亚组NF-kBp65显著高于Non-CALs亚组[(22.86±3.11)vs.(13.55±3.23)(P<0.05)].KD组NF-kBp65抑制物IKBa明显低于感染对照组及健康对照组[(5.69±2.03)vs.(21.22±4.37,28.58±7.89)(P<0.001)].CALs亚组中胞核NF-KBP65/IkBα比值与CALs的严重程度呈正相关(r=0.536,P<0.05).(2)KD组TNF-αmRNA、MCP-1mRNA较两对照组明显升高[(7.02±2.91)vs.(3.05±2.33,0.61±0.38)(P<0.01);(3.89±1.10)vs.(1.11±0.42.0.04±0.01)(P<0.01)]CALs亚组TNF-αmRNA、MCP-1mRNA明显高于Non-CALs组[(8.79±1.52)vs.(6.13±2.52)(P<0.05);(4.26±0.78)vs.(3.01±0.53)(P<0.05)].结论 NF-kBp65信号通路的活化参与KD急性期血管炎的发生机制,可能参与冠状动脉损害的形成.     

p53 mRNA和Caspase-3 mRNA在幼年大鼠心肌缺血再灌注损伤中的动态表达及药物干预作用

目的 探讨p53 mRNA、Caspase-3 mRNA在幼年大鼠心肌缺血再灌注损伤(MIRI)不同时间的动态表达与心肌细胞凋亡的关系,观察药物干预的影响.方法 将70只幼年大鼠随机分为3组:对照组10只,模型组和治疗组各30只,制备MIRI模型.对照组和模型组大鼠右股静脉注入9 g/L盐水,治疗组注入甲泼尼龙针,原位杂交检测p53、Caspase-3 mRNA.原位末端标记染色(TUNEL)检测凋亡细胞在各组不同时间的动态变化.结果 模型组与治疗组p53 mRNA表达峰值出现在缺血再灌注(IR)8 h,Caspase-3 mRNA峰值在IR 12 h;模型组凋亡细胞在IR 12 h达高峰,治疗组延迟至IR 24 h;相同时间点治疗组与模型组比较,p53 mRNA和Caspase-3 mRNA表达均显著减弱(Pa<0.05),凋亡细胞数显著减少(P<0.05).结论 p53和Caspase-3通过诱发心肌细胞凋亡在MIRI的病程中发挥重要作用,甲泼尼龙干预对MIRI具有一定的积极作用.     

吲哚胺2,3-双加氧酶在哮喘小鼠模型中的作用

目的 探讨吲哚胺2,3-双加氧酶(IDO)在哮喘小鼠模型中所起的作用.方法 卵白蛋白(OVA)致敏、激发诱导哮喘小鼠模型.分别检测肺组织中IDO在蛋白水平和mRNA水平上的表达.免疫荧光组织化学方法检测树突状细胞(DCs)在肺组织中的分布和成熟状态.结果 ①OVA诱导和激发的小鼠哮喘模型的症状和肺部炎症病理改变较对照组严重;哮喘组小鼠血清总IgE为(165.50 ± 30.13)ng/ml,对照组为(94.45 ± 28.30)ng/ml,差异有统计学意义(P ＜ 0.05).②哮喘组小鼠肺部IDO的表达低于对照组.以平均累积光密度为指标检测的小鼠肺组织中IDO在蛋白水平上的表达,哮喘组为11.38 ± 6.05,对照组为23.62 ± 8.92,差异有统计学意义(P ＜ 0.05);实时荧光定量PCR检测小鼠肺组织中IDO在mRNA水平上的表达,哮喘组是对照组的33%,显著低于对照组(P ＜ 0.05).③小鼠肺组织中CD11c+CD86+细胞主要分布于肺泡壁和小血管周围.以双标阳性细胞平均累积荧光强度为指标检测小鼠肺组织中CD11c+CD86+细胞数量,哮喘组的中位数为9 961.86(7 406.52 ～ 12 724.98),对照组为15 974.60(10 006.39 ～ 16 171.46),差异有统计学意义(P ＜ 0.05).结论 哮喘小鼠肺组织表达IDO低,且成熟树突状细胞减少.提示在哮喘小鼠中,由于成熟的树突状细胞较少,产生IDO偏少,这可能在哮喘发生中起着重要作用.

Abstract：

Objective To investigate the role of indoleamine 2,3-dioxygenase(IDO)in asthma.Methods The mouse asthma model was induced by ovalbumin(OVA).IDO expression was detected on the level of protein and mRNA respectively.Distribution and maturation of dendritic cells were detected by the immunofluorescence method.Results (1)The symptoms and lung inflammation in the model group were more serious than control group.The serum total IgE was significantly higher in the model group than that in the control group,165.50 ± 30.13 ng/ml vs.94.45 ± 28.30 ng/ml(P ＜ 0.05).(2)IDO expression in the model group was lower than that in control group.On the level of protein,mean intergrated optical density was 11.38 ± 6.05 in the model group vs.23.62 ± 8.92 in the control group(P ＜ 0.05);on the level of mRNA,IDO expression of the model group was 33% of the control group(P ＜ 0.05).(3)CD11c+CD86+ cells were distributed in alveolar wall and around small vessels.The quantity of CD11c+CD86+ cells in lungs of the model group were significantly smaller than that in the control group.The median intergrated fluorescence intensity was 9961.86(range,7 406.52 ～ 12 724.98)in the model group vs.15974.60(range,10 006.39 ～ 16 171.46)in the control group(P ＜ 0.05).Conclusions IDO expression is low and matured dendritic cells are less in situ in the asthma model.These suggest that less matured DCs may produce less IDO,which may play an important role in asthma.     

Breastfeeding perceptions in communities in Mangochi district in Malawi

Aim: To investigate mothers’ perceptions of breastfeeding and influences from their social network. Methods: A cross‐sectional survey was carried out in Mangochi district, Malawi where questionnaire data from 157 rural and 192 semi‐urban mother–infant pairs were obtained. Results: The proportion of mothers who thought that exclusive breastfeeding should last for 6 months and those who reported to have actually exclusively breastfed were 40.1% and 7.5% respectively. Of those who reported practising exclusive breastfeeding for 6 months, 77.5% stated that exclusive breastfeeding should last for 6 months. This opinion was independently associated with giving birth in a Baby‐Friendly facility, OR = 5.22; 95% CI (1.92–14.16). Among the mothers who thought that exclusive breastfeeding should last for less than 6 months, 43.9% reported having been influenced in their opinion by health workers. Infant crying was the most common (62.4%) reason for stopping exclusive breastfeeding. Conclusion: The findings illustrate the positive impact health workers can have, as well as the need to raise awareness of the benefits of exclusive breastfeeding among both health workers and mothers. Furthermore, continued counselling of mothers on how to deal with stressful infant behaviour such as crying may assist to prolong exclusive breastfeeding.     

胶原成分在儿童系膜增生性肾小球肾炎中的变化

目的观察系膜增生性肾小球肾炎（ＭｓＰＧＮ）系膜区胶原成分的变化。方法应用链菌素亲生物素过氧化酶连接法观察了３０例轻度ＭｓＰＧＮ肾穿刺活组织标本和正常的肾小球系膜区Ⅳ型胶原及其α链（α１、α３、α５链）、Ⅵ型胶原及Ⅰ型胶原的变化。结果（１）正常肾脏组织中，Ⅳ型胶原及其α１（Ⅳ）链分布于系膜区和基底膜，α３（Ⅳ）、α５（Ⅳ）链分布于基底膜，Ⅵ型胶原分布于系膜区、肾小球基底膜和间质，Ⅰ型胶原仅分布于肾间质。（２）在轻度ＭｓＰＧＮ时，系膜区内Ⅳ型胶原及其α１链、Ⅵ型胶原含量较正常对照明显增多（Ｐ＜０．０１）；当系膜区系膜细胞超过４个时，Ⅰ型胶原开始在肾小球内出现，且在硬化肾小球内Ⅰ型胶原均呈阳性；α３（Ⅳ）、α５（Ⅳ）链与正常对照比较无明显变化，硬化肾小球α３（Ⅳ）、α５（Ⅳ）染色呈阳性。结论系膜区胶原成分增多可先于系膜细胞增生，并随系膜细胞增生而增多，间质胶原成分Ⅰ型胶原，不但出现于硬化肾小球内，而且出现于系膜细胞增生较重时     

Cardiomyopathy in children in Ahmedabad

A clinical study and follow up of 20 children with cardiomyopathies upto age of 16 years are presented. The DCM was most common variety followed by RCM and HCM in pediatric age group. SHMD presenting with cardiomyopathy were common in infancy and early childhood. Cardiomyopathies presented most frequently between 2–5 years and 10–16 years age group with DCM having almost equal distribution. Clinical presentation of various types is described, despite of vigorous decongestive and vasodilator treatment in advanced cases, course was rapidly downhill and prognosis is poor in general.     

Achalasia in siblings in infancy

Achalasia is rare in children, more so familial. We report two siblings with familial achalasia who presented in their infancy with vomiting and failure to thrive. Achalasia can be misdiagnosed as upper gastrointestinal obstruction as happened in one of our siblings. Esophageal contrast roentgenography is diagnostic. Both the children were treated successfully by transabdominal esophagomyotomy with fundoplication.     

北京地区2007年儿童A群轮状病毒腹泻的流行病学研究

目的 研究北京地区急性腹泻儿童中A群轮状病毒(RV)感染的流行病学特点.方法 收集2007年4月至12月我院肠道门诊就诊的2039例急性腹泻患儿的粪便标本,采用标记金的A群RV单克隆抗体,以免疫层析双抗体夹心法定性检测A群RV抗原.结果 2039份粪便标本中,621份检测到A群RV,总检出率为30.5%(621/2039),其中男430例(69.2%),女191例(30.8%).RV感染者中,以6个月～2岁年龄段的患儿为最多,共571例(91.9%).检出率以10～12月份最高,均在30%以上,其中高峰出现在11月份,达43.4%.北京地区18个区县的统计数据显示,距市区较近的区县RV抗原检出率较低,边远区县较高.少部分患儿合并肠道细菌感染.结论 A群RV为北京地区2岁以下儿童急性腹泻病的主病原,6个月～2岁婴幼儿是A群RV的易感人群,10～12月份为北京地区的流行高峰.在流行季节对肠道细菌感染患儿常规进行A群RV抗原检测有助于避免漏诊和进行更合理的治疗.     

哮喘大鼠气道重塑中尾加压素-的表达变化

目的：研究哮喘大鼠气道重塑血清和支气管肺泡灌洗液（BALF）中尾加压素 Ⅱ（U-II）含量的变化及其作用。方法：32只雄性Sprague-Dawley大鼠随机分为正常对照组、哮喘2周组、哮喘4周组和哮喘8周组,每组8只。以卵清白蛋白(OVA)致敏与激发建立哮喘大鼠气道重塑模型,图像分析技术测量大鼠支气管壁总面积和平滑肌面积,计算单位基底膜周径(Pbm)的支气管壁厚度(Wat)和平滑肌厚度(Wam),ELISA法测定血清和BALF中U-II的含量。结果：哮喘各组Wat及Wam均明显高于正常对照组（P<0.01）;哮喘组血清和BALF中U-II含量均显著高于正常对照组（P<0.01）,其中哮喘8周组血清和BALF中U-II含量显著高于哮喘4周组和哮喘2周组（P<0.01）,哮喘4周组也显著高于哮喘2周组（P<0.01）。各组大鼠BALF中的U-II含量与Wat及Wam呈正相关,BALF与血清中U-II含量亦呈正相关。结论：哮喘大鼠气道重塑血清和BALF中U-II含量增加;且U-II含量的变化与气道重塑相关。［中国当代儿科杂志,2010,12（4）：287-289］     

转流术治疗小儿精索静脉曲张

目的 采用转流术治疗小儿精索静脉曲张，重新建立精索静脉通道，使静脉回流受阻立即得到改善，消除因睾丸淤血而造成的损害，以利睾丸的正常发育。方法 对28例30侧(左侧26例，双侧2例)精索静脉曲张与腹壁下静脉进行吻合，通过腹壁下静脉，髂静脉转流，手术在放大镜下应用显微外科技术进行，其中28侧用精索静脉主干，2侧结扎一条属支，用另一条静脉进行吻合。结果 通畅率为100％。术后扩张迂曲静脉团消失，阴囊下坠感消失。术后随访24例，时间为3个月-10年。除1例二次手术证实为一条静脉属支漏扎而复发外，另23例全部治愈。结论 精索静脉曲张转流术效果明显优于结扎术，可减少因睾丸淤血对其造成的进一步损害，且术后复发率低。     

Poisoning in children in Japan

The Japan Poison Information Centre (JPIC) received 31510 inquiries about poisoning in children under 6 years old being exposed to poison in the fiscal year 1995. The most frequently implicated products were tobacco (20%) and the peak age for ingestion of household products was 1 year and younger (83.3%). Especially, the inquiries related to children less than 1 year old were 35.7% of the cases. In contrast, the American Association of Poison Control Centers (AAPCC) data showed that the most common poisonings were due to pharmaceutical products and the inquiries related to children less than 1 year old were only 12.1%. The objective of this report was to find out the poison exposure in children in Japan and to compare the data with that of AAPCC.     

Diagnosis of Chlamydia pneumoniae in community-acquired pneumonia in children in Chile

The incidence of Chlamydia pneumoniae in community-acquired pneumonia in children was studied prospectively in 112 children aged 1 mo to 14 y. Diagnosis of C. pneumoniae was performed by polymerase chain reaction (PCR) on nasopharyngeal aspirates and serology by the microimmunofluorescence test on a single serum specimen. Three (2.7%) cases of pneumonia due to this agent were diagnosed by both PCR and serology. C. pneumoniae was not found in any of 62 children below 5 y of age. In the age group 5-8 y, only 1/30 (3%) was found positive, and in the age group 9-14 y, C. pneumoniae was diagnosed in 2/20 (10%) children. Conclusion: Although the number of enrolled patients is small, and the diagnostic techniques used may have some limitations, the results of this study suggest that C. pneumoniae plays a minor role in the aetiology of pneumonia in children less than 9 y of age in our country. However, it should be considered as a potential agent in pneumonia in older children.