裸鼠皮下移植人神经母细胞瘤模型的建立和HIF-1α及其相关蛋白的表达

目的 探讨应用人神经母细胞瘤细胞株建立裸鼠皮下神经母细胞瘤模型,测定缺氧诱导因子(hypoxia-inducible factor-1,HIF-1α)及其相关蛋白在该模型中的表达,了解HIF-1α及其相关蛋白在神经母细胞瘤发生发展中的作用,为神经母细胞瘤(neuroblastoma NB)的发病机制研究和诊治提供新的思路与方法.方法 应用KP-N-NS人肾上腺神经母细胞瘤细胞株建立模型.采用S-P免疫组织化学方法检测裸小鼠皮下移植瘤、手术切除的神经母细胞瘤肿瘤标本、正常人肾上腺髓质组织中HIF-1α、VEGF及TGF-α的表达情况.结合免疫组化结果对HIF-1α阳性组与HIF-1α阴性组裸鼠的体重变化、瘤体重量、存活天数进行比较,并对实验结果进行统计学处理.结果 4周后解剖结果证实成模率80%;HIF-1α、VEGF及TGF-α在肿瘤组织中的阳性表达率显著高于正常组织(P＜0.01),肿瘤组与阳性对照组HIF-1α、VEGF、TGF-α三种蛋白的阳性染色的积分光密度(integrating optical density,IOD)值明显高于阴性对照组(P＜0.01).HIF-1α在肿瘤组与阳性对照组中的表达与VEGF的表达成正相关(P＜0.05),与TGF-α的表达无明显相关性(P＞0.05);HIF-1α阳性组与HIF-1α阴性组体重变化、瘤体重量及存活天数比较,差异有统计学意义(P＜0.01);HIF-1α阳性组较HIF-1α阴性组裸鼠生存时间短(P＜0.05).结论 该模型用于研究人类神经母细胞瘤,造模时间短,成瘤率高,生物学行为与人类神经母细胞瘤接近.HIF-1α、VEGF及TGF-α蛋白的阳性表达与神经母细胞瘤的浸润及转移有关.HIF-1α与VEGF在神经母细胞瘤的发生发展过程中可能存在一定的协同作用.HIF-1α的高表达是预后不良的一个重要指标.

Abstract：

Objective To study the HIF-1αand its related proteins' expression in an athymic mouse human neuroblastoma xenograft model. Methods The was established by subcutaneous injection of human adrenal neuroblastoma cells. The expressions of HIF-1α, VEGF and TGF-α were detected by S-P immunohistochemical techniques. The tumor group was divided into HIF-1α positive group and the HIF-1α negative group according to the immunohistochemistry results. The body weight,tumor weight and survival days of the HIF-1αpositive mice and HIF-1αnegative mice were statistically analyzed. Results Four weeks later, significant neuroblastoma growth occurred in 40 mice (80%).The positive HIF-1α, VEGF and TGF-α rate of the tumor group was (73. 70 ± 10. 68) %, (69. 80 ±9. 91) %, and (71.43 ± 8. 52) 0% , respectively. It is significantly increased compared with the control group (P＜0. 01 ), which were (9. 67 ± 4. 53) %, (6. 80 ± 3. 40) %, and (6. 50 ± 4. 44) %, respectively. The integrated optical density (IOD) of HIF-1α, VEGF and TGF-α was 141.97 ± 7. 98,151.85 ±14. 35, and 139. 94 ± 4. 50 in tumor group, and 141.34 ± 6. 44, 144. 06 ± 8. 51 and 149. 00 ± 13. 63 in positive control group. They were significantly higher than those of the negative control group (P＜0. 01 ). In the tumor group and positive control group, the expression of HIF-1α was positively correlated with the expression of VEGF (x2 = 7. 778,r= 0. 504,P＜0. 01). However, no correlation between the expression of HIF-1α and TGF-α was found(x2 = 0. 208,r= 0. 934,P＞0. 05). The body weight,tumor weight and survival days of HIF-1α positive mice were significantly different with those of HIF-1α negative mice (P＜0. 01 ). The mice with positive HIF-1α expression had shorter survival time than the mice with negative HIF-1α (x2 = 19. 013,P＜0. 05). Conclusions The athymic mouse human neuroblastoma xenograft model is a good model to research the biology behavior of human neuroblastoma.The overexpression of HIF-1α, VEGF and TGF-αwere found in neuroblastoma which may play important roles in tumor invasion and metastasis. HIF-1αis synergistic with VEGF to promote carcinogenesis. The overexpression of HIF-1αis correlated with poor prognosis.     

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.     

儿童肾母细胞瘤HMGB1、RAGE及VEGF的表达及意义

目的 探讨高迁移率族蛋白-1(HMGB1)、晚期糖基化终末产物受体(RAGE)和血管内皮生长因子(VEGF)在小儿肾母细胞瘤组织中的表达及其在肿瘤血管浸润、淋巴结转移中的意义.方法 收集郑州大学第一附属医院2003年1月至2010年1月小儿外科手术切除经病理证实为肾母细胞瘤的石蜡标本50例、15例同期相应瘤旁组织和7例正常肾组织标本.应用免疫组织化学SP染色辅以计算机图像分析系统分析结果的方法,研究HMGB1、RAGE及VEGF在各组标本中的表达及意义.结果 HMGB1、RAGE在肾母细胞瘤组织中的表达(144.46±13.55、138.18±12.26)与两者在瘤旁组织和正常肾组织中表达(107.47±5.27、103.91±4.29;100.98±4.82、100.82±3.32)相比差异均有统计学意义(P＜0.05),HMGB1在瘤旁与正常肾之间表达差异有统计学意义(P＜0.05).VEGF在瘤体组与瘤旁组中(147.57±13.77,140.28±7.85)和在正常肾组织组中的表达(106.38±1.92)相比差异显著(P＜0.05).三者在临床分期Ⅲ～Ⅳ期和预后不良组织型组及淋巴结转移组的肾母细胞瘤组织中的表达(148.69±12.17、147.73±6.71、163.14±7.50;157.88±4.44、155.29±3.97、169.17±4.42;152.11±7.36、151.56±5.46、163.83±7.20)显著高于临床分期Ⅰ～Ⅱ期和预后良好组织型及无淋巴结转移组(P＜0.05)(139.23±5.83、133.30±11.23、140.85±8.87;139.52±5.22、135.39±9.71、143.94±10.39;138.61±5.59、133.00±8.75、141.18±8.95).相关分析显示肾母细胞瘤组织中HMGB1与RAGE、VEGF的表达均成正相关(r=0.424,P=0.002;r=0.453,P=0.001),RAGE与VEGF之间无明显相关(r=0.237,P=0.101).结论 HMGB1、RAGE和VEGF的高表达与肾母细胞瘤的临床分期、病理类型、淋巴结转移有关,参与了肿瘤血管浸润及淋巴结转移,HMGB1/RAGE可能是促进肾母细胞瘤转移的一条信号通路,给我们靶向治疗肾母细胞瘤提供了新的思路.

Abstract：

Objective This study aim to assess the expressions of high mobility group box chromosomal proteinl ( HMGB1) and receptor foradvanced glycation end products (RAGE) and vascular endothelial growth factor (VEGF) in Wilms'tumor and their clinical significance both in the tumor blood vessel infiltrates and in the lymph node metastasis. Methods Fifty cases of Wilms'tumor samples, which had been confirmed by pathology, were collected from The First Affiliated Hospitals of Zhengzhou University from january 2003 to january 2010. And 15 cases were taken from the adjacent kidney tissues at the same time. Other 7 cases were normal kidney tissues. The expression of HMGB1, RAGE and VEGF were detected by the Immunohistochemical SP staining in each group of specimens. The expression intensity was analyzed by computer image processing and their significance in each group of specimens were studied. Results The expressions of HMGB1 and RAGE in the Wilms'tumor tissues (144. 46 ± 13. 55、138.18 ±12. 26) were compared with those in the adjacent kidney tissues and in the normal kidney tissues( 107. 47 ± 5. 27、103. 91 ± 4. 29; 100. 98 ± 4. 82、 100. 82 ± 3. 32). The expressions of HMGB1 in the adjacent kidney tissues and in the normal kidney showed significant differences. There was a significant differences between the Wilms' tumor group and the adjacent kidney tissues or in the normal kidney tissues ( F ＜ 0.05 ). The expressions of VEGF proteins in Wilms'tumor tissues and in the adjacent kidney tissuess(147. 57 ± 13. 77,140. 28 ± 7. 85) comoared to those in the normal kidnev tissues(106. 38 ± 1. 92)were remarkably different(P＜0. 05). The expressions of HMGB1 ,RAGE and VEGF proteins in Wilms' tumor tissues of stage Ⅲ-Ⅳ and high risk histopathology and lymph node metastasis group (148. 69 ± 12.17、147. 73 ± 6. 71、163.14 ± 7. 50; 157. 88 ± 4. 44、155. 29 ± 3. 97、169. 17±4.42;152. 11 ± 7. 36、151. 56 ± 5. 46、163. 83 ± 7. 20)were significantly higher than those of stage Ⅰ-Ⅱ and low risk histopathology and no lymph node metastasis group(P＜0. 05) (139. 23 ± 5. 83、133. 30 ± 11. 23、140. 85 ± 8. 87; 139. 52 ± 5. 22、135. 39 ± 9. 71、143. 94 ± 10. 39; 138. 61 ± 5. 59、133. 00 ±8. 75、141.18 ± 8. 95). There was a positive correlation between the expressions of HMGB1 with RAGE and VEGF(r = 0. 424, P = 0. 002; r = 0. 453, P = 0. 001), but the correlation between the RAGE and VEGF is not clear (r = 0. 237, P = 0. 101). Conclusions There was a high expression of HMGB1 ,RAGE and VEGF in the Wilms'tumor tissues and they are partly related to the clinical stages and pathological type and lymph node metastasis of Wilms'tumor. They perhaps participated in the tumor blood vessel infiltration and the lymph node metastasis and the pathway of HMGB1/RAGE is perhaps the main mechanism correlated with the metastasis of Wilms'tumor.     

RNA干扰趋化因子受体4基因表达对神经母细胞瘤体外侵袭能力的影响

目的 构建趋化因子受体4(CXCR4)小分子干扰RNA(siRNA)表达载体,研究其对体外神经母细胞瘤侵袭能力的影响.方法 选择CXCR4高表达的神经母细胞瘤SH-SY5Y细胞系,设计合成人CXCR4基因不同靶点的能编码siRNA的3条双链DNA序列,克隆到真核表达载体pSilencerTMneo中构建siRNA表达载体,体外脂质体介导转染SH-SY5Y细胞,用半定量RT-PCR分析CXCR4基因mRNA的变化,用免疫组织化学和Western blot分析CXCR4蛋白表达,Transwell小室检测细胞的侵袭能力.结果 成功构建了CXCR4-siRNA表达载体,转染后半定量RT-PCR检测神经母细胞瘤细胞CXCR4 mRNA丰度分别为siR1转染组0.32±0.09、siR2转染组0.35±0.13和siR3转染组0.33±0.11,相对于对照组0.58±0.13表达下降(P＜0.05);转染后免疫组化检测神经母细胞瘤细胞CXCR4的蛋白表达分别为siR1转染组75.98±4.81、siR2转染组75.52±3.95和siR3转染组76.35±6.51,相对于对照组92.196±3.89表达下降(P＜0.01);转染后Western b1ot检测神经母细胞瘤细胞CXCR4的蛋白表达分别为siR1转染组0.1103±0.0023、siR2转染组0.1203±0.015和siR3转染组0.1308±0.0018,相对于对照组0.4832±0.0012表达下降(P＜0.01);且转染后神经母细胞瘤细胞侵袭能力较对照组53.11±3.72降低(P＜0.05),siR1转染组为25.48±2.81、siR2转染组为30.89±2.77、siR3转染组为18.83±1.79.结论 CXCR4-siRNA表达载体通过降低CXCR4基因的蛋白表达能显著抑制神经母细胞瘤细胞的体外侵袭能力,有望为神经母细胞瘤的基因治疗开辟新途径.

Abstract：

Objective To explore the effect of silencing chemokine receptor type 4 (CXCR4) by siRNA on the invasion capability of neuroblastoma cell line SH-SY5Y in vitro. Methods Three siRNAs targeting CXCR4 were chemically synthesized and transfected into SH-SY5Y cells. The transfection efficiency was observed under fluorescence microscope. CXCR4 expression at mRNA and protein levels were detected by semi-quantitative RT-PCR and Western blotting. The invasion capability of the cells was evaluated by Boyden Chamber in vitro. Results Compared with control groups, after the SH-SY5Y cells being transfeeted with the three CXCR4 targeting siRNAs, CXCR4 mRNA in transfected cells significantly decreased (0. 32 ± 0. 09, 0. 35 ± 0. 13 and 0. 33 ± 0. 11 vs 0. 58 ± 0. 13, P＜0. 05 ), CXCR4 protein detected by immunohistochemistry was decreased (75. 98 ± 4. 81, 75. 52 ± 3. 95and 76. 35 ± 6. 51 vs 92. 196 ± 3. 89, P＜0. 01 ), CXCR4 protein detected by Western blotting was also decreased (0. 1103 ± 0. 0023, 0. 1203 ± 0. 0015 and 0. 1308 ± 0. 0018 vs 0. 4832 ± 0. 0012, P＜0. 01 ).The invasion capability of the SH-SY5Y cells was decreased 48 hours after the cells were transfected (25.48±2.81, 30.89±2.77 and 18.83± 1.79 vs 53. 11 ±3.72, P＜0.05). Conclusions Silencing CXCR4 by siRNA decreases the invasion capability of SH-SY5Y cells.     

腹腔镜下高位肛门闭锁肛门成形术及中期临床评估

目的 评估腹腔镜下高位肛门闭锁成形术后临床疗效和直肠肛门功能.方法 2004年6月至2007年9月收治高位肛门闭锁患儿61例,33例行腹腔镜下肛门成形术(LAARP),平均年龄5.3个月;28例行后矢状入路肛门成形术(PSARP),平均年龄4.9个月.随访包括手术时间、住院时间和并发症.手术后3～4年对患儿进行排便功能的Kelly评分(KCS)、磁共振成像(MRI)和直肠肛管向量测压(AVVM)评估.结果 LAARP和PSARP组手术时间分别为(112.5±12.4)min和(120.4±18.5)min(P＞0.05),LAARP组住院时间(11.3±2.1)d短于PSARP组(14.6±2.3)d(P＜0.01).两组患儿KCS无显著差异(3.52±1.42比3.49±0.82,P＞0.05).MRI显示:LAARP组33例患儿中在I线上和M线上各有1例存在直肠位置偏移;PSARP组28例患儿中I线上有4例存在直肠位置偏移,M线上有3例存在直肠位置偏移.直肠肛管向量测压结果显示:与PSARP组相比,LAARP组非对称指数小,向量容积大,静息时和收缩时肛管压力高(P＜0.05).但高压带长度(15.2±5.8比15.1±6.2 mm)和直肠肛管抑制反射阳性率(84.8%比85.7%)无显著差异.结论 高位肛门闭锁患儿LAARP术后排便控制满意,与PSARP相比,LAARP术后住院时间短、直肠位置更准确.长期随访对评估LAARP术后功能非常必要.

Abstract：

Objective To evaluate the clinical outcomes and postoperative anal function in infants with congenital high imperforate anus who underwent laparoscopically assisted anorectal pullthrough (LAARP). Methods From January 2004 to July 2007,33 consecutive patients (28 males and 5 females,age ranging from 3 to 10 months) with high imperforate anus underwent LAARP. Clinical data of the LAARP group were retrospectively compared with those treated by posterior sagittal anorectoplasty ( PSARP,n = 28) at the same time period. Anorectal function of these patients was evaluated using the Kelly's score,anorectal vector volume manometry(AWM) and magnetic resonance imaging (MRI) at the age of 3. 1 to 4. 4 years. Results The operative time in LAARP and PSARP groups was 112. 5 ± 12.4 and 120.4 ± 18.5 min (P ＞ 0. 05), respectively. The length of hospital stay in LAARP group was shorter than that of PSARP group (11. 3 ± 2. 1 vs. 14. 6 ± 2. 3 days,P＜0. 01). No significant difference was observed between LAARP and PSARP groups regarding Kelly's score (3. 52 ± 1. 42 vs. 3. 49 ± 0. 82). Although MRI revealed the lower rate of poorly aligned rectum in LAARP group than PSARP group at both I-line (3. 0% vs. 14. 3%) and M-line (3. 0% vs. 10. 7%) levels,no statistically significant difference was noted (P＞0. 05). Compared with the PSARP group, lower asymmetric index, larger vector volume, and higher anal canal pressure at rest and during voluntary squeeze were observed in LAARP group (P＜0. 05), without significant differences in the length of high-pressure zone (15. 2 ± 5. 8 vs. 15. 1 ± 6. 2 mm) and the presence of rectoanal relaxation reflex (84. 8% vs. 85.7%). Conclusions Satisfactory fecal continence can be achieved in patients with high/intermediate type imperforate anus after LAARP. LAARP has some advantages over PSARP, including shorter hospital stay and better position of rectum. However, long-term follow-up is necessary to compare the benefits of LAARP against PSARP.     

高效抗逆转录病毒治疗对艾滋病患儿CD8+T细胞活化分子CD38和人类白细胞抗原DR表达水平的影响及其与病毒载量的关系

目的 观察高效抗逆转录病毒治疗(HAART)对我国艾滋病患儿CD8+T细胞活化标志分子CD38和人类白细胞抗原DR(HLA-DR)表达水平的影响及其与病毒载量的关系.方法 对194例接受HAART的艾滋病患儿进行横断面研究,用流式细胞术检测CD4+、CD8+T细胞数,以及CD8+/CD38+和CD8+/HLA-DR+T细胞比例,RT-PCR检测血浆HIV RNA载量.并检测52名健康儿童CD8+/CD38+和CD8+/HLA-DR+T细胞比例作为正常对照.结果 本组中,135例病毒载量＜400 copies/ml,59例病毒载量≥400 copies/ml.病毒载量≥400 copies/ml组CD8+/CD38+T和CD8+/HLA-DR+T细胞水平显著高于病毒载量＜400 copies/ml组,差异有统计学意义(29.6±10.1 vs19.9±9.8;17.7±6.4 vs 9.6±6.1,P＜0.05);病毒载量＜400 copies/ml组,CD8+/CD38+T细胞接近正常水平,而CD8+/HLA-DR+T细胞仍高于正常水平,差异有统计学意义(19.9±9.8 vs 15.6±9.0;9.6±6.1 vs 5.8±3.3,P＜0.05).CD8+/CD38+和CD8+/HLA-DR+T细胞百分比均与病毒载量成正相关(前者相关系数R=0.403,P=0.03,后者相关系数R=0.569,P=0.09).结论 艾滋病患儿CD8+T细胞活化程度与病毒载量成正比,有效的HAART治疗能够显著地降低HIV感染者免疫活化程度,CD8+/CD38+和CD8+/HLA-DR+T细胞百分比可能是资源有限地区替代病毒载量检测的潜在指标.

Abstract：

Objective To study the expression of CD38 and HLA-DR on CD8 + T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART) and the relationship of immune activation and disease progression. Methods A cross-section study of 194 pediatric AIDS patients receiving HAART was carried out and 52 age-matched healthy children were recruited as control. The percentage of CD4+ , CD8+ , CD8+/CD38 + and CD8+/HLA-DR+ T cells was tested using flow cytometry, and HIVRNA in plasma was detected by quantitative RT-PCR. Results One hundred and ninety-four pediatric AIDS patients were divided into two groups according to the viral load: 59 patients with VL≥400 copies/ml and 135 patients with VL＜400 copies/ml. The percentage of CD8 +/CD38+ and CD8 +/HLA-DR+ T cells of patients with VL≥400 copies/ml was significantly higher than that of patients with VL ＜ 400 copies/ml (P ＜ 0. 05 ). Of patients with VL ＜ 400 copies/ml, the percentage of CD8 +/CD38 + T cells was nearly normal, and the percentage of CD8 +/HLA-DR+ T cells was higher than normal level ( P ＜ 0. 05 ). There was a positive correlation between percentage of CD8+/CD38+ and of CD8 +/HLA-DR+ T cells and viral load ( R = 0. 403, P = 0. 03 for the former and R = 0. 569, P = 0. 09 for the later). Conclusions Effective HAART could decrease immune activation of HIV-infected children significantly. And there was a positive correlation between percentage of CD8 +/CD38 + and of CD8 +/HLA-DR + T cells and viral load, suggesting that the two indicators might be used as the substitution of viral load in resource-limited areas.     

胆道闭锁多中心综合诊断治疗方案研究

目的 探讨胆道闭锁综合诊疗方案应用的有效性和安全性.方法 根据具有地域代表性的四家儿童医院专家确定的胆道闭锁诊疗方案,2007年1月开始对胆道闭锁进行诊断及治疗.收集2004年1月至2006年12月四家医院经确诊为胆道闭锁并进行根治手术的病例作为对照组2007年1月至2009年12月病例作为观察组.使用标准化量表统计首次就诊年龄、手术年龄、术前及术后3个月,6个月和1年黄疸清除率,术后胆管炎发生率,2年自体肝生存率.结果 符合诊断标准498例,实施Kasai术456例,其中对照组184例,手术168例;观察组314例,手术288例.二组首次就诊年龄无明显差异[(33±27)d比(34±26)d,P＞0.05],使用标准化诊断方案后平均手术年龄由(64±24)d下降为(74±31)d,P＜0.05;观察组术后3个月、6个月及1年退黄率(53.9%、56.9%、58.0%)较对照组(38.9%、39.9%、43.4%)明显提高(P＜0.05);其术后胆管炎发生率亦显著降低(31.8%比47.8%,P＜0.05);对照组2年自体肝生存率39.3%,观察组56.1%(P＜0.05).结论 标准化胆道闭锁综合诊断治疗方案可降低手术年龄、提高术后黄疸清除率、减少术后反流性胆管炎,显著提高术后2年自体肝生存率.

Abstract：

Objective To evaluate the efficacy and safety of a standardized protocol for the diagnosis and treatment of biliary atresia (BA) in children.Methods This study enrolled 498 patients from 4 hospitals diagnosed with BA from January 2004 to December 2009.The BA patients of the standardized protocol group,who enrolled in this study after January 2007,were diagnosed and treated using the standardized protocol.The rest of the patients were taken as controls.The patients were followed up for 2 years.The age of the first diagnosis,the age at operation,jaundice clearance,the occurrence of cholangitis and two year survival rates with native liver were retrospectively reviewed and compared between the 2 groups.Results Among the 498 patients,456 patients underwent Kasai procedure.Age of the first diagnosis between the two groups showed no difference (33 ± 27 vs 34 ± 26 days,P＞0.05),but the patients of the standardized protocol group received surgeries at earlier ages (64 ± 24 vs 74 ± 31 days,P＜0.05).The jaundice clearance rates of 3,6 and 12 months after operation in the patients of the standardize protocol group were higher than those of controls (53.9%,56.9%,58.0% vs 38.9%,39.9%,43.4%,P＜0.05).Patients of the standard protocol group had lower occurrence of cholangitis (31.8% vs 47.8%,P＜0.05),and better two years survival rate (56.1% vs 39.3%,P＜0.05).Conclusions The standardized protocol for the diagnoses and treatment of BA improves the clinical outcomes of the children with BA.     

小儿肾母细胞瘤中SOCS-1和SOCS-3的表达及其临床意义

目的 探讨小儿肾母细胞瘤中细胞因子信号转导抑制因子-1(SOCS-1)和细胞因子信号转导抑制因子-3(SOCS-3)的表达,以及与小儿肾母细胞瘤不同病理类型之间的内在关系.方法 应用RT-PCR法及Western-Blot法测定10例小儿正常肾脏组织、30例小儿肾母细胞瘤及瘤旁组织中sOCS-1和SOCS-3的表达.结果 与正常小儿肾脏组织(mRNA表达:0.721±0.183、0.694±0.148,蛋白表达:0.746±0.168、0.652±0.183)相比,肾母细胞瘤组织中SOCS-1和SOCS-3 mRNA表达(0.156±0.027、0.103±0.021)及蛋白的表达(0.205±0.079、0.185±0.041)均明显降低(P＜0.01).在肿瘤不同病理类型之间SOCS-1和SOCS-3mRNA的表达(预后良好型:0.175±0.086、0.149±0.100,预后不良型:0.128±0.074、0.091±0.067)有差异(P＜0.05).瘤旁组织SOCS-1 和SOCS-3mRNA的表达(0.572±0.154、0.315±0.137)及蛋白的表达(0.628±0.183、0.439±0.137)与正常肾脏组织有差异(P＜0.05).结论 SOCS-1和SOCS-3的表达下降可能与小儿肾母细胞瘤的发生、发展密切相关,与病理分期也有一定联系.

Abstract：

Objective To investigate the expression levels of suppressor of cytokine signaling-1 and suppressor of cytokine signaling-3 in the Wilms' tumor and to study the inherent relationship between the different pathological types. Methods The expression levels of SOCS-1 and SOCS-3 were assessed by RT-PCR and Western-Blot in ten normal kidney tissues, thirty nephroblastoma specimens and adjacent tissues. Results Compared to the normal kidney tissues(mRNA expressions: 0.721±0.183、0.694±0.148,protein expressions:0.746±0.168、0.652±0.183), the expressions of mRNA (0.156±0.027、0.103±0.021)and protein(0.205±0.079、0.185±0.041)of the SOCS-1 and SOCS-3 in nephroblastoma were significantly lower (P＜0.01). There were statistically significant between mRNA expressions and the different pathological types(FH:0.175±0.086,0、149±0.100,UH:0. 128±0.074、0.091±0.067) (P＜0.05).The difference betweenneuroblastoma and the normal tissues in expressions of mRNA (0. 572 ± 0. 154、0. 315 ± 0. 137)and protein(0. 628± 0. 183、0. 439 ± 0. 137)of the SOCS-1 and SOCS-3 were statistically significant (P＜0. 05). Conclusions The low SOCS-1 and SOCS-3 expressions may have a role in the development and progress of nephroblastoma, and the different pathological types.     

B16F10黑色素瘤小鼠双阴性T细胞变化及与肿瘤生长关系的研究

目的 探讨B16F10黑色素瘤小鼠双阴性T细胞(DNT)数量变化及其与肿瘤生长的关系.方法 以B16F10-luc-G5细胞系接种Babl/C小鼠建立移植瘤模型,并以活体成像系统(IVIS)监测其生长动态,流式细胞术检测接种后不同时间外周血中T淋巴细胞的比例;流式分选正常小鼠DNT再回输到B16F10黑色素瘤荷瘤小鼠体内,观察DNT对移植瘤生长的作用.结果 ①荷瘤鼠DNT含量高于正常鼠(P<0.05);CD4~+细胞含量与肿瘤大小高度负相关(R~2=0.999 3);②以外源性DNT干预移植瘤,DNT干预组移植瘤生长速度较对照组快(P<0.05).结论 B16F10黑色素瘤小鼠体内DNT的数量升高且与肿瘤大小正相关,且提高体内DNT含量可促进移植瘤生长,提示DNT将是肿瘤免疫治疗的靶点之一.     

2009甲型H1N1流感患儿免疫功能改变初探

目的 探讨2009甲型H1N1流感(以下简称"甲流")患儿免疫功能及可能的免疫发病机制.方法 深圳市儿童医院2009年11月1日-2010年1月10日甲流住院患儿60例,轻症35例(轻症肺炎),重症25例(重症肺炎或甲流相关性脑病,死亡3例),同年龄正常对照组20例.采用real-time PCR、流式细胞术及ELISA检测外周血单个核细胞胞浆模式识别受体(PRRs)维甲酸诱导基因I/黑色素瘤分化相关基因5(RIG/MDA5)、胞膜PRRs Toll样受体(TLRs)分子及其信号途径传导分子、细胞因子/趋化因子及负性调节因子变化;T、B及NK细胞凋亡及凋亡相关基因TRAIL和CASPASE-3表达.结果 (1)甲流患儿RIG/MDA5表达、TLR2、TLR4表达明显高于正常对照组[TLR2(9.69±3.15)×10-2vs.(3.96±0.83)×10-2,t=10.16,P＜0.05;TLR4(10.23±2.85)×10-2vs.(7.46±2.18)×10-2,t=3.76,P＜0.05],以重症甲流患儿增高为著,RIG/MDA5表达增高最为明显;TLRs途径信号传导分子MyD88、TRAM等表达明显高于轻症甲流患儿.(2)甲流患儿CD3+[(1.22±0.38)×109/Lvs.(3.59±1.10)×109/L,t=9.21,P＜0.05]、CD4+、CD8+T细胞及NK细胞绝对计数明显低于正常对照组,B细胞无明显改变.(3)轻症甲流患儿TNF-α、IL-6、IL-1β等炎症细胞因子血浓度或基因高于正常对照组,重症患儿炎症细胞因子TNF-α[(6.42±1.76)×10-2vs.(9.05±2.51)×10-2,t=4.55,P＜0.05]明显低于正常对照组.IFN-α/β表达持续高于正常对照组,尤以重症甲流患儿为著;IFN-I诱导基因IP-10[(20.52±6.09)×10-2vs(1.18±0.34)×10-2,t=18.74,P＜0.05]、RANTES或iNOS轻症患儿表达高于正常对照组,重症患儿表达则趋于减少.(4)甲流患儿CD3+[(32.90±7.66)%vs.(20.21±6.58)%,t=6.21,P＜0.05]、CD4+、CD8+T细胞、NK细胞凋亡高于正常对照组,以重症患儿更为显著.凋亡相关基因TRAIL和CASPASE-3表达明显高于正常对照组.(5)重症患儿PRRs负性凋节因子SOCS1、SOCS3、IRAK-M、TRAF4及FLN29表达明显高于轻症患儿,抗炎细胞因子IL-10及IL-10/TNFα比值随病情加重增高.结论 甲流患儿机体免疫功能紊乱,轻症患儿处于全身免疫激活状态,重症患儿同时存在免疫激活/免疫抑制反应.     

α-干扰素对血管瘤血管内皮细胞周期、凋亡的影响

目的 观察α-干扰素对体外培养血管瘤内皮细胞周期、凋亡的影响,探讨干扰素治疗血管瘤的机制.方法 用组织块法培养增生期血管瘤内皮细胞,分别用1×104U,1×105U,1×106Uα-干扰素干预,对照组不加任何药物,继续培养24h后,用流式细胞仪检测细胞周期、凋亡变化.结果 α-干扰素作用24 h后血管瘤内皮细胞G1期前出现二倍体凋亡峰;1×104U、1×105U、1×106Uα-干扰素组G1期百分率分别为(63.68±0.66)%、(72.97±1.56)%、(63.27±5.64)%,对照组为(58.75±0.32)%,与对照组比较,P＜0.05;1×104U、1×105U、1×106Uα-干扰素组S期百分率分别为(32.99±2.01)%、(25.02±1.31)%、(37.93±5.31)%,对照组为(26.46±1.94)%,与对照组比较,P＞0.05;1×104U、1×105U、1×106Uα-干扰素组G2/M期百分率分别为(0.01±0.05)%、(2.43±0.27)%、(3.99±1.27)%,其百分率随α-干扰素浓度升高而增高.细胞凋亡百分率随α-干扰素浓度增加而增高,1×104U、1×105U、1×106Uα-干扰素组凋亡百分率分别为(11.89±0.56)%、(18.88±3.04)%、(31.92±1.92)%,对照组凋亡率为(3.25±0.23)%,与对照组比较,P＜0.01.结论 α-干扰素能导致血管瘤血管内皮细胞G1、G2期阻滞和凋亡,细胞凋亡率与药物浓度存在剂量依赖关系.干扰素可能通过血管内皮细胞G1、G2期阻滞,抑制血管瘤血管内皮细胞的增殖、诱导血管内皮细胞凋亡而达到治疗血管瘤的作用.     

体位引流辅助气管吸引对胎粪吸入综合征的防治效果

目的 评价体位引流辅助气管吸引防治胎粪吸入综合征(MAS)的临床效果.方法 2007年1月至2008年12月我院收治的窒息合并MAS新生儿61例,分为对照组24例,生后气管插管应用胎粪吸引管气管内吸引;观察组37例,插管后体位引流再行气管吸引.主要观察两组气管内吸出胎粪量、MAS并发症及转归.结果 观察组气管内抽出胎粪量(2.16±1.82)ml,较对照组(1. 23±0.97)ml明显增多,差异有显著性(P＜0.05);观察组吸净胎粪所需插管次数为(1.19±0.46)次,较对照组[(1.79±0.83)次]显著减少,差异有显著性(P＜0.01).观察组并发症发生率为8.11%,明显低于对照组29.17%,差异有显著性(P＜0.05).观察组吸氧时间(包括机械通气)为(21.30±22.38)h,较对照组[(52.91±39.20)h]显著缩短,差异有显著性(P＜0.01);观察组住院天数(9.24±3.94)d,对照组为(14.39±6.49)d,差异有显著性(P＜0.01).观察组病死率为0,对照组病死率4.17%,差异无显著性(P＞0.05).观察组1 min Apgar评分与对照组相近,差异无显著性(P＞0.05);观察组5 minApgar评分8～10分者占70.16%,明显高于对照组(58.34%),差异有显著性(P＜0.05).结论 体位引流辅助气管吸引可有效清除气管内胎粪,减少MAS并发症的发生,缩短吸氧时间和住院天数.     

儿童肿瘤化疗后粒细胞减少期并发ARDS肺泡巨噬细胞活性检测

目的 研究儿童肿瘤化疗后粒细胞减少期并发感染后出现ARDS,肺泡巨噬细胞(AM)HLA-DR的表达情况.方法 收集13例儿童肿瘤化疗后并发ARDS患者,其中5例为粒细胞减少期,予粒细胞集落刺激因子治疗.所有患儿行纤维支气管镜检查,收集支气管肺泡灌洗液(BALF)进行细胞计数、分类,流式细胞仪检测AM HLA-DR表达率.结果 粒细胞减少组ARDS患儿肺泡细胞总数、AM绝对值和AM百分比分别为(62.6±9.6)/μl、(40.8±4.3)/μl和65.9%±9.0%,粒细胞正常组ARDS患儿分别为(124.0±6.7)/μl、(67.6±10.7)/μl和54.6%±8.7%,两组差异均有显著性(P＜0.05).粒细胞减少组BALF中AM HLA-DR表达率较粒细胞正常组明显下降(35.3%±5.8%vs62.2%±5.8%),差异有非常显著性(P＜0.01).结论 粒细胞减少的ARDS患儿呈"肺泡细胞减少症"状态,AM呈失活状态.     

电针可下调缺氧缺血性脑损伤幼鼠大脑皮层硫化氢含量

目的 建立缺氧缺血性脑损伤(HIBD)大鼠模型,研究电针干预后大鼠大脑皮层硫化氢(H2S)含量及相关酶的表达变化,探讨电针治疗神经系统疾病的气体生物学机制.方法 SD大鼠32只,日龄7 d,随机分为4组,假手术(sham)对照组,sham+电针(EA)组,HIBD对照组和HIBD+EA组,每组8只.Sham+EA组、HIBD+EA组大鼠于造模次日开始予以电针刺激百会、大椎穴,30min/d,14 d为1疗程.对照组仅固定四肢,不予针刺.疗程结束次日将大鼠处死、取材.应用敏感硫电极法测定大脑皮层H2S含量;应用免疫组织化学法定位和半定量检测胱硫醚-β-合成酶(cystathionine-β-synthase,CBS)表达;应用蛋白质免疫印迹技术定量分析大脑皮层组织CBS表达.结果 HIBD对照组幼鼠脑皮层H2S含量为(26.83±4.31)nmol/mg蛋白,较sham对照组[(22.78±1.54)nmol/mg蛋白]明显升高,差异有统计学意义(P＜0.01);HIBD+EA组幼鼠脑皮层H2S含量为(18.08±2.71)amol/mg蛋白,sham+EA组为(18.91±2.78)amol/mg蛋白,较相应对照组明显降低,差异有统计学意义(P＜0.01);EA组大鼠脑皮层CBS表达较相应对照组明显降低,差异有统计学意义(P＜0.05).结论 电针可下调大脑皮层H2S/CBS体系,这可能是电针发挥对脑损伤修复作用的机制之一.

Abstract：

Objective To establish hypoxic-ischemic brain damaged (HIBD) rat model,investigate whether H2S and cystathionine-β-synthase (CBS), the key enzyme for its generation, may be a mediator of electro-acupuncture(EA) stimulation treatment for HIBD. Methods Thirty-two healthy Sprague-Dawley neonatal rats were divided into four groups randomly: sham control group ( n = 8 ); sham + EA group ( n =8); HIBD control group ( n = 8); and HIBD + EA group ( n = 8 ). HIBD rat models were established on their 7-day-old. From the next day ,rats of sham + EA group and HIBD + EA group were electric stimulated 30 min daily for 14 d,BAIHUI and DAZHUI as the acupoints. Control ones were just fixed at the same time,without acupuncture. The rats were sacrificed on the 22 nd day, one day after the treatment course. Cortical H2S concentration was measured by sensitive sulphur electrode assay. The CBS protein expression was measured by western blot analysis and immunohistochemistry for localization. Results The concentration of cortical H2S in HIBD control group was (26. 83 ± 4. 31 ) nmol/mg protein, which was significantly higher than that of sham control group[(22. 78 ± 1.54) nmol/mg protein]( P ＜ 0. 01 ). The H2 S levels in HIBD + EA group and sham + EA group were ( 18.08 ± 2.71 ) nmol/mg protein and ( 18.91 ± 2. 78 ) nmol/mg protein, respectively. Compared with corresponding control group, they were much lower( P ＜ 0. 01 ). The expression of CBS protein in rats with EA stimulation decreased in cortex compared to corresponding control group( P ＜0. 05 ).Conclusion EA can down-resulate H2S/CBS pathway. This may be one of the mechanisms of how EA contributes to the recovery of brain damage.     

MDR1基因转染的骨髓单个核细胞对放疗后造血功能重建的研究

目的 研究BALB/c小鼠放射治疗后经尾静脉回输携带多药耐药MDR1基因的骨髓单个核细胞(bone marrow mononuclear cells,BM-MNCs)对造血功能重建的影响.方法 BAB/c近交系小鼠32只,随机分为4组:正常对照组、空白对照组、阴性对照组和实验组.正常对照组不做任何处理,其他3组首先接受1.5Gy60Co-γ射线全身照射,空白对照组尾静脉回输等量生理盐水,阴性对照组经尾静脉回输未转染MDR1基因BM-MNCs,实验组回输已转染MDR1基因BM-MNCs.动态观察各组外周血变化.结果 同种异体回输后第七天,阴性对照组和实验组与空白对照组比较均表现为白细胞恢复提前.实验组7、10、14 d白细胞计数分别为(2.9±0.3)×109/L、(3.2±0.2)×109/L、(4.2±0.3)×109/L,阴性对照组为(2.7±0.2)×109/L、(2.8±0.2)×109/L、(3.5±0.3)×109/L,实验组与阴性对照组比较,白细胞数量增加,差异有统计学意义(t=2.21、3.53、4.73,P＜0.05),14 d后差异无统计学意义(t=0.79,P＞0.05).结论 同种异体携MDR1基因的骨髓单个核细胞移植,能在早期显著提高外周血白细胞数量,利于骨髓造血微环境恢复,加快辐射损伤后骨髓早期造血功能重建.     

血管紧张素Ⅱ对幼鼠输尿管梗阻后AQP2表达变化的调控

目的 通过观察血管紧张素Ⅱ(angiotensin Ⅱ,ANG Ⅱ受体阻滞剂坎地沙坦对双侧输尿管梗阻(bilateral ureteral obstruction,BUO)幼鼠肾脏水通道蛋白2(aquaporin 2,AQP2)表达的影响,探讨ANG Ⅱ对梗阻肾脏功能和AQP2的调控作用。方法 24只慕尼黑幼鼠随机分为BUO组、坎地沙坦干预组(BUO+ CAN)和对照组(Sham),每组n=8只。BUO组和BUO+ CAN组均行双侧输尿管结扎,并采用微型泵分别给以生理盐水和坎地沙坦,Sham组仅游离输尿管但不结扎,24h后解除梗阻并继续观察48h后收集血液和肾脏标本,采用免疫印记技术检测肾脏AQP2表达水平。结果 梗阻解除后BUO组与Sham组相比尿量显著增高,(92±7)μl·min-1 ·kg-1比(25±3)μl· min-1·kg-1、尿渗透压显著降低,(636±55) mosmol/kgH2O比(1 853±163) mosmol/kgH2O、血浆渗透压和血浆醛固酮均显著增高,分别为(336±10) mosmol/kgH2O比(303±7)mosmol/kgH2O和(4.1±0.2)nmol/L比(1.4±0.1)nmol/L;肾脏AQP2表达下调到Sham组17％各组比较差异有统计学意义,P＜0.05。BUO+ CAN组与BUO组相比尿量显著减少,(55±5)μl·min-1 ·kg-1比(92±7)μl·min-1 ·kg-1,尿渗透压显著增高(783±47) mosmol/kgH2O比(636±55) mosmol/kgH2O,血浆醛固酮含量显著降低(2.8±0.5) nmol/L比(4.1±0.2)nmol/L,肾脏AQP2表达增高,各组比较差异有统计学意义,P＜0.05。结论 ANG Ⅱ受体拮抗剂可通过阻止AQP2下调纠正水代谢紊乱,保护肾功能,提示ANG Ⅱ通过调节肾脏AQP2表达参与输尿管梗阻后肾脏水代谢变化。