谷氨酰胺对坏死性小肠结肠炎新生大鼠Toll样受体4的调控

目的：研究坏死性小肠结肠炎（NEC）新生大鼠肠道组织中Toll样受体4（TLR 4）和caspase-3的表达情况,探讨谷氨酰胺（Gln）对NEC肠道保护作用的可能机制。方法：60只早产新生大鼠随机分为模型组、Gln组和对照组（n=20）。代乳品人工喂养+缺氧+冷刺激建立NEC模型,Gln组采用代乳品+Gln（0.3 g/kg）人工喂养,同时给予缺氧冷刺激,对照组未进行任何干预,第3天处死后取出肠道,苏木素-伊红染色后对回肠进行病理评分,免疫组织化学检测空肠、回肠及结肠caspase-3、TLR-4蛋白的表达,实时荧光定量PCR检测空肠、回肠及结肠TLR-4 mRNA的表达。结果：与对照组相比,模型组的病理评分、caspase-3、TLR-4蛋白和TLR-4 mRNA的表达均增加（P＜0.01）;与模型组相比,Gln组的各项指标均有所下降（P＜0.05）。结论：TLR-4可能参与了NEC的发病机制,降低TLR-4的表达,抑制肠黏膜细胞的凋亡可能是Gln保护NEC新生大鼠肠道的机制之一。     

孕酮对新生鼠脑室周围白质软化模型脑白质少突细胞标记物O1 和O4 表达的影响

目的探讨孕酮对新生鼠脑室周围白质软化(PVL)动物模型脑室周围白质O1和O4表达的影响。方法 2日龄新生SD大鼠随机分为模型组、实验组、假手术组。模型组和实验组结扎左侧颈总动脉并吸入低氧气源(8%O2+92%N2)0.5 h制备PVL动物模型,实验组脑缺氧缺血后立即腹腔注射孕酮10 mg/(kg·d);假手术组只分离左侧颈总动脉不结扎、不缺氧。术后1、4、7、14天比较三组脑组织病理改变,免疫组织化学染色方法检测三组大鼠各个时点脑室旁白质区O1、O4表达。结果假手术组各时点脑白质区未见异常病理改变;模型组和实验组均出现左侧脑室扩大和脑室周围白质软化灶等病理改变,但实验组各时点脑白质病理改变均明显轻于模型组。三组大鼠脑室旁白质区O1、O4阳性细胞的积分光密度(IOD)在术后1、4、7天的表达逐渐增加,第7天达高峰,第14天下降,差异有统计学意义(P均0.01)。在术后1、4、7、14天,三组脑室旁白质区O 1、O 4阳性细胞IOD在同一时间点均以假手术组最多,实验组其次,模型组最少,差异有统计学意义(P0.01)。结论孕酮可减轻新生大鼠PVL模型动物脑室旁白质区病理损伤,促进脑室周围白质O1、O4表达,促进少突胶质细胞分化成熟。     

脑白质损伤新生大鼠脑白质P75NTR蛋白与RhoA mRNA的表达及意义

目的：最近研究发现NgR-P75NTR- RhoA信号通路在神经损伤和重塑方面发挥关键作用,但其确切传导机制及在缺氧缺血后新生动物P75NTR对下游分子RhoA调控变化尚不清楚。该实验在建立新生大鼠缺氧缺血性脑白质损伤(WMD)模型基础上,观察WMD新生大鼠脑白质P75NTR和RhoA mRNA表达变化, 探讨WMD时两者的关系及意义,为临床治疗早产儿WMD提供新思路和实验依据。方法：制备新生大鼠WMD模型,光镜及电镜观察脑组织形态学改变。应用免疫组织化学方法及荧光定量RT-PCR检测对照组及WMD 12,24,48,72 h和7 d组病变侧脑白质P75NTR蛋白及RhoA mRNA表达变化。结果：光镜和电镜形态学发现WMD组缺氧缺血48 h后即有显著的脑室周围脑白质损伤。WMD组大鼠纹状体和胼胝体P75NTR蛋白水平12 h开始升高,48 h达峰值,至72 h略有下降,与对照组比较差异有显著性,均P<0.01,之后下降明显,第7天与对照组比较差异无统计学意义（P>0.05）。WMD组大鼠脑白质RhoA mRNA 表达12 h开始升高,48 h达到峰值,与对照组比较,差异有显著性,均P<0.05。72 h仍高于对照组水平,之后开始下降,第7天与对照组比较差异无显著性（P>0.05）。结论：WMD新生大鼠脑白质中P75NTR蛋白水平呈现先上升后下降的趋势。其升高与介导神经细胞凋亡和抑制神经轴突再生可能有关,表达下降则与缺氧缺血性损伤加重神经细胞坏死相关。RhoA mRNA的表达变化趋势与P75NTR一致,提示P75NTR水平升高可促使RhoA mRNA高表达。［中国当代儿科杂志,2007,9（4）：317-320］     

新生期大鼠反复癎性发作对海马神经发生的影响

目的 探讨新生期大鼠反复痫性发作对海马神经发生的影响。方法64只生后1d的Wistar大鼠随机分为惊厥组40只和对照组24只。惊厥组新生鼠在生后1d（P1）、4d（P4）、7d（P7）给予腹腔注射匹罗卡品,制备新生鼠癫痫模型,对照组腹腔注射生理盐水。惊厥组分别在第3次致痫后即刻、第4、14、45天4个时间点处死,各时间点设相应对照组,处死前36h给予惊厥组和对照组大鼠腹腔注射BrdU,用BrdU标记新生细胞,再以NF200双标免疫荧光法标记神经元。结果对照组BrdU标记阳性细胞呈年龄依赖性变化,不同年龄组BrdU阳性细胞的差异有统计学意义（P〈0．01）,惊厥组3次发作后即刻和第4天齿状回BrdU阳性细胞数明显少于相应日龄对照组（P〈0．01）,而发作后14d和45dBrdU阳性细胞数高于相应日龄对照组,发作后14d差异无统计学意义。各时间点惊厥组和对照组大多数BrdU阳性细胞同时表达NF200。结论正常大鼠的神经发生有年龄依赖性,随着日龄增大,齿状回颗粒细胞增生减慢,而新生期反复痫性发作对大鼠不同时期影响不同,反复发作会导致幼鼠早期神经发生减少,而后期神经发生增加,这些新生细胞不仅存在于齿状回,还可见于CA3区及海马以外的某些区域,并可分化为神经元。     

肺炎链球菌脑膜炎室管膜下区神经发生的实验观察

目的观察内源性神经干细胞在幼年大鼠肺炎链球菌性脑膜炎室管膜下区的增殖。方法 3周龄SD大鼠分为感染组(n=28)、对照组(n=16)和假手术组(n=16)。感染组脑池穿刺接种肺炎链球菌,对照组注射生理盐水,假手术组仅剪开皮肤,分别于接种后24 h,及5、14及21天各处死4只大鼠,采用免疫组化法检测神经干细胞间接标志物5-溴脱氧尿核苷(Brdu)的表达,观察内源性神经干细胞在室管膜下区的增殖情况。结果感染组、对照组及假手术组大鼠在接种后第24小时、5天、14天及21天时,双侧室管膜下区均可见Brdu阳性细胞,感染组的Brdu阳性细胞在接种后24小时开始增加,第5天达到高峰,随后逐渐下降,各时间点的差异有统计学意义(P0.05)。在接种后24小时、5天和14天时,感染组的Brdu阳性细胞数均高于对照组及假手术组,差异有统计学意义(P0.05);接种后第21天时,感染组与对照组的差异无统计学意义(P0.05),但与假手术组的差异仍有统计学意义(P0.05)。结论肺炎链球菌性脑膜炎可刺激幼年大鼠室管膜下区神经干细胞的增殖。     

SOX9在胆道闭锁小鼠模型中的表达特点

目的检测胆道闭锁小鼠模型中肝脏SOX9阳性细胞及SOX9/HNF4α双阳细胞表达特点及其与肝纤维化进展之间的关系。方法取新生的71只BALB/c小鼠进行实验, 在出生24～28 h内, 新生的57只BALB/c小鼠行20 μl RRV腹腔注射作为实验组, 其余14只同时注射同体积生理盐水作为对照组, 实验组和对照组根据注射后的时间又各自按随机数字表法分为7天组(实验组8只、对照组5只)、10天组(实验组19只、对照组5只)及14天组(实验组30只、对照组4只), 观察小鼠临床特征变化。在注射后7 d、10 d、14 d分别留取相应组别小鼠肝脏标本, 对肝脏组织切片进行苏木精-伊红染色及天狼猩红染色以评估肝纤维化程度, 同时利用多重免疫荧光染色对肝脏组织切片进行SOX9阳性细胞及SOX9/HNF4α双阳细胞计数。利用t检验及One-way ANOVA进行数据分析。结果最终7天组5只小鼠造模成功, 10天组6只小鼠造模成功, 14天组4只小鼠造模成功, 对照组小鼠14只均顺利存活。胆道闭锁小鼠皮肤黄染且体重增长缓慢;所有小鼠肝脏均存在SOX9阳性细胞, 胆道闭锁小鼠肝脏中央静脉及小叶间静脉...     

胰岛素样生长因子-1减少胎羊缺血性脑白质损伤后淀粉样前体蛋白表达(英文)

目的：淀粉样前体蛋白 (β APP)是脑白质损伤早期敏感的指标 ,并参与缺氧缺血性脑损伤机制。本研究观察胎羊缺血性脑白质损伤及胰岛素样生长因子 1(IGF 1)治疗对淀粉样前体蛋白 (β APP)表达的影响。方法：胎羊于胎龄 117 12 4天 (足月为 14 7天 )时通过双侧颈动脉阻塞 30min造成双侧脑缺血损伤 ,损伤后胎羊随机分为损伤组 (n =8)和重组人IGF 1(rhIGF 1)治疗组 (n =9) ;另设正常对照组 (n =5 ) ,为假手术动物。治疗组缺血后 90min经侧脑室注射 3μgrhIGF 1;损伤组经侧脑室注射等量人工脑脊液。缺血损伤后 96h结束实验 ,处死动物 ,取出胎羊 ,固定脑组织。免疫组化法检测脑白质胶质原纤维酸性蛋白 (GFAP)、β APP阳性细胞及白质内髓鞘碱性蛋白 (MBP)密度。应用免疫荧光双标记观察APP表达阳性细胞。结果：与正常对照组 (2 7.8± 4 .8)比较 ,缺血损伤组MBP密度 (4.7± 7.1,P <0 .0 0 1)明显减少。正常对照组未见 β APP阳性细胞 ,损伤后阳性细胞数明显增加 (49.6± 2 3.7,P <0 .0 0 1) ,rhIGF 1治疗可减少 β APP阳性细胞数 (17.9± 16 .5 ,P <0 .0 1)。免疫荧光双标记显示部分细胞为 β APP GFAP双标阳性细胞。 结论：胎羊缺血性脑白质损伤可导致星形胶质细胞表达β APP ,β APP表达增加可能与脑损伤有关     

咪康唑对大鼠缺氧缺血性早产儿脑白质损伤的保护作用

目的探讨咪康唑对早产儿脑白质损伤(WMD)大鼠髓鞘的保护作用。方法新生3日龄SD大鼠随机分为假手术组、WMD模型组、10 mg/(kg·d))和40 mg/(kg·d)咪康唑组,每组15只;采用结扎右侧颈总动脉,缺氧80 min的方法制作早产儿WMD模型。咪康唑组于建模后第1~5天腹腔注射10 mg/(kg·d)和40 mg/(kg·d)咪康唑,WMD模型组注射等浓度二甲基亚砜(DMSO)。采用髓鞘碱性蛋白(MBP)免疫荧光染色及Western blot检测脑白质特异性MBP表达量,超微结构电镜观察髓鞘超微结构变化,并比较各组幼鼠体质量变化。结果 WMD大鼠经咪康唑治疗后,胼胝体MBP表达量较WMD模型组高,差异有统计学意义(P0.05)。咪康唑治疗组MBP的表达量较模型对照组增高。模型对照组胼胝体髓鞘疏松,髓鞘内小空泡形成,呈筛网状改变,髓鞘厚度明显降低,结构紊乱。经咪康唑治疗后可明显改善缺氧缺血所致的脱髓鞘改变。WMD模型组幼鼠体质量增长速度较假手术组明显减慢,咪康唑治疗后大鼠的体质量生长速度增快。结论咪康唑可通过促进髓鞘形成保护新生大鼠脑缺氧缺血诱导的白质损伤,并改善大鼠的生长发育情况。     

丙氨酰-谷氨酰胺对低出生体重新生大鼠缺氧性肠损伤后胰岛素样生长因子1的影响

目的　探讨丙氨酰-谷氨酰胺(Ala-Gln)对低出生体重(LBW)新生大鼠缺氧后肠组织胰岛素样生长因子-1(IGF-1)及IGF-1受体(IGF-1R)水平的影响。方法　将孕鼠分别置于吸烟和非吸烟环境中饲养, 非吸烟条件下饲养孕鼠所分娩的新生大鼠设为A组; 将吸烟条件下饲养孕鼠所分娩的正常体重新生大鼠设为B组, LBW新生大鼠随机分为对照组(C组)、缺氧-复氧组(D组)和Ala-Gln组(E组); 每组24只。D、E组行缺氧-复氧操作连续3 d, 每天2次造模, E组在每日缺氧处理前腹腔注射Ala-Gln(10 mL/kg), C、D组以等量生理盐水行替代注射。各组于生后第4、7、10天分别处死8只大鼠取肠组织。采用ELISA法检测各组肠组织IGF-1水平; 免疫组化法检测各组肠组织IGF-1R水平。结果　A组与B组IGF-1及IGF-1R蛋白水平在各时间点差异无统计学意义(P>0.05)。C组IGF-1及IGF-1R蛋白水平呈上升趋势, 第7天时均较其他各组升高(P<0.05), 第10天时趋于正常水平, 与A、B组比较差异无统计学意义(P>0.05)。D组各时间点IGF-1及IGF-1R蛋白水平均明显低于C组(P<0.05)。E组在第4、7天IGF-1及IGF-1R蛋白水平低于C组(P<0.05), 但到第10天上升至接近C组水平, 且显著高于D组水平(P>0.05)。结论 宫内和生后缺氧易使LBW新生大鼠出现肠损伤, 肠道外给予大剂量Ala-Gln可减轻LBW新生大鼠缺氧性肠损伤程度, 对缺氧性肠损伤有一定保护作用。     

早期脑损伤新生大鼠脑白质神经蛋白聚糖和多功能蛋白聚糖表达及意义

目的观察宫内感染致脑损伤仔鼠脑白质Neurocan和Versican表达情况。方法 40只受孕母鼠随机分为对照组(n=10)和脂多糖组(n=30),给予受孕18 d的脂多糖组孕鼠脂多糖(血清型055)380μg/kg,连续2 d腹腔注射;对照组孕鼠腹腔注射同等剂量的生理盐水。每组随机选取幼鼠各60只,于0,1,7,14,21,28日龄各处死10只仔鼠,HE染色法检测脑组织病理变化,荧光定量RT-PCR方法测定Neurocan和Versican相对表达量。结果对照组孕鼠无死亡及提前分娩,共娩出活产足月仔鼠124只;脂多糖组中孕鼠5只死亡,6只提前分娩,剩余19只共娩出活产足月仔鼠132只,死亡24只。与对照组比较,0,1,7,14,21,28日龄脂多糖组新生大鼠脑白质Neurocan mRNA和Versican mRNA表达增加,差异有统计学意义(P<0.05)。结论宫内感染致脑损伤后,脑白质区Neurocan和Versican的表达明显上调。     

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.     

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］     

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.     

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.     

Rotavirus infection in children in Japan

Currently, a high morbidity of rotavirus diarrhea has been seen in children in developed and developing countries. Improvement of the vaccines is necessary in order to reduce the burden of diarrhea caused by rotavirus. A survey of rotavirus infection from diarrheal stool specimens in children of seven regions in Japan was conducted from 1984 to 1999. The present study discusses the survey results and reviews the national and international data of more than 23 papers and congress proceedings about rotavirus infection in Japan. We analyze the prevalence of rotavirus infection in acute diarrheal in- and outpatients, the distribution of rotavirus G-serotypes and surveillance data for seasonality and age groups in Japan. The data indicated that rotavirus is the most important cause of diarrhea in Japan among young children, with the prevalence ranging from approximately 9.7 to 88%. The most common rotavirus strains belonged to serotype G1, specifically since 1993. Serotypes G2, G3 and G4 had also been documented to be predominantly based in the area and year before 1992. However, untypeable rotavirus strains had been found each year, with a prevalence up to 56.7% which suggests that rare serotypes (except G1-4) or new serotypes might exist. Unexpectedly, in Tokyo and Sapporo from 1998 to 1999, G9 was found to be the first most prevailing serotype with a high prevalence of 52.9 and 71.4%, respectively. Despite these data from different geographic areas, the year under investigation was relatively clear in respect to seasonality, with a peak of rotavirus activity in late winter (February) through early spring (March). Age distribution had also characterized that the infection was predominant among children aged 1-2 years of age, although it was also common in children of 2-3 years. In addition, mixed infection with bacteria was documented.