胎羊单侧输尿管不完全性梗阻后肾脏足细胞表型转化及PAX2、VEGF的表达

Objective To investigate the phenotypic change of the podocytes and paired box gene 2 (PAX2) and vascular endothelial growth factor (VEGF) expressions in fetal lambs' kidneys with unilateral partial ureteral obstruction. Methods Sixteen fetal lambs were randomly assigned to the obstruction and control group. The obstruction group included 12 lambs, whose superior segment ureters were tied with splited silastic tube to induce unilateral partial ureteral obstruction between 75and 85 gestational days. The control group had 4 fetal lambs subjected to sham operation. After birth,the kidneys of the lambs were harvested to study the phenotypic change of the podocytes as well as expressions of PAX2 and VEGF in the kidneys. Results Nine out of the 12 lambs of the obstruction group were delivered alive. All 4 sham operated lambs were delivered alive. Renal cortex cysts of varying sizes, interstitial fibrosis, a decrease in glomeruli number, and severe podocyte foot process fusion (4. 20± 1.08% vs 86. 79 ± 1.66%) were found in the obstructed kidneys compared with the control kidneys. The PAX2 expression in obstructed kidneys was significantly higher than that of the control kidneys (1.43 ± 0. 09 vs 2. 44 ± 0. 09, P＜0. 05). The VEGF expression was significantly decreased compared with the control kidneys (0. 80 ± 0. 15 vs 0. 33 ± 0. 14, P＜0. 05). Conclusions The changes of the phenotype and PAX2 and VEGF expressions may play roles in the pathogenesis of obstructive nephropathy.     

胎羊单侧输尿管不完全性梗阻解除前后肾脏组织足细胞表型转化及PAX2、VEGF的表达

目的 从超微结构和分子学方面探讨胎儿期单侧输尿管不完全性梗阻解除后肾脏恢复之机制.方法 26只孕75～85 d的胎羊分3组:梗阻组,对12只胎羊实行宫内手术造成单侧输尿管不完全性梗阻;梗阻解除组,对10只胎羊以同样的手术方法造成单侧输尿管不完全性梗阻,3周后,再次宫内手术解除梗阻;对照组,对4只胎羊实行宫内手术但不结扎输尿管.待胎羊出生后,取其肾脏标本检测病理改变,足细胞表型转化以及PAX2和VEGF的表达变化.结果 梗阻组12只胎羊,其中9只顺利出生;梗阻解除组10只胎羊,其中8只顺利出生;对照组4只胎羊,均顺利生产.与对照组羔羊肾脏相比,梗阻组羔羊梗阻侧肾脏,表现为皮质囊性改变,间质纤维化,肾小球数目显著减少;足细胞足突广泛融合[(4.20±1.08)%比(86.79±1.66)%];PAX2表达显著升高(1.43±0.09比2.44±0.09);而VEGF表达明显减少(0.80±0.15比0.33±0.14).解除梗阻后,肾脏的囊性改变大大改善,肾小球数目增加,但细胞外基质沉着仍明显;足细胞足突融合减轻[(41.18±3.13)%比(86.79±1.66)%];VEGF的表达显著升高(2.08±0.21比0.33±0.14);PAX2的表达也有一定程度的恢复(2.05±0.14比2.44±0.09).结论 通过对胎羊单侧输尿管不完全性梗阻解除前后肾脏组织病理改变、足细胞表型转化以及PAX2和VEGF表达变化的观察,说明宫内干预能减缓甚至逆转输尿管不完全性梗阻所致的肾脏损伤性改变.

Abstract：

Objective To investigate ultrastructure and molecular change of kidneys after correction of partial unilateral ureteral obstruction. Methods The 26 fetal lambs at 75-85 days of gestation were divided into 3 groups. Obstructed group: partial unilateral ureteral obstruction was achieved with intrauterine surgery in 12 fetal lambs. Decompressed group: partial unilateral ureteral obstruction in 10 fetal lambs first was released 3 weeks later by intrauterine surgery. Control group: a sham operation was performed. The kidneys of the lambs were removed after birth to study the transformation of podocytes as well as expression of paired-box 2 (PAX2) and vascular endothelial growth factor (VEGF). Results Overall, 21 fetal lambs survived: 9 from the obstructed group, 8 from the decompressed group, and 4 from the control group. Compared with the control kidneys, the obstructed kidneys showed cysts of various sizes in the cortex, fibrosis of the interstitial tissue, significantly decreased number of glomeruli, severer podocyte foot process fusion (4. 20 ± 1. 08% vs 86.79 ± 1. 66%) , markedly increased PAX2 (1. 43 ± 0. 09 vs 2. 44 ± 0. 09) and decreased VEGF expressions (0. 80 ± 0. 15 vs 0. 33 ± 0. 14). However, in the decompressed kidneys, the cystic changes decreased, number of glomeruli increased, but extracellular matrix (ECM) deposition was still obvious, fusion of the podocyte foot processes was reduced (41. 18±3. 13% vs 86. 79± 1. 66%), VEGF expression was significantly increased (2. 08 ± 0. 21 vs 0. 33 ± 0. 14) , and expression of PAX2 was restored to some extent (2. 05 ± 0. 14 vs 2. 44 ± 0. 09) in decompressed kidneys. Conclusions Relief of obstruction in utero can reverse the development of nephropathy, evidenced by the phenotype transformation of podocytes, expression of PAX2 and VEGF.     

单侧隐睾大鼠对侧睾丸组织中SCF/c-kit基因表达变化及意义

Objective To investigate the pathologic changes and expressions of SCF and c-kit in the contralateral testes in rat model of unilateral cryptorchidism. Methods Thirty male SD rats were maintained under controlled temperature and constant photoperiodic conditions with access to food and water. The rats were randomly assaigned to the control group and the experimental unilateral cryptorchidism group. The left testis of the rats in the unilateral cryptorchidism group was placed into the abdominal cavity. The control group rats were subjected to sham surgery. Three months later, the rats were sacrificed and their right testes were harvested. The pathological changes were observed under microscope. The mRNA and protein expression of SCF and c-kit were also investigated using quantitative Real-time RT-PCR, Western blotting and immunohistochemical staining. Apoptotic germ cells were detected by TUNEL staining. Results All rats survived to the endpoints. The right testes of rats of the unilateral cryptorichid group were smaller than those of the control group. Normal testes of the control group rats manifested active spermatogenesis and orderly arrangement of germ cells.However, disordered and sloughed germinal cells with less distinct seminiferous tubule borders were observed in the contralateral testes of rats with experimental unilateral cryptorichid under the microscope. Cmpared with the control testes, the mRNA and protein expression of SCF and c-kit of the contralateral testes of rats with experimental unilateral cryptorichid was decreased (P＜0.05). The apoptotic germ cells were increased (19.7±3.83 vs 5. 4 ± 1.02, P＜0. 05). The SCF/c-kit expression was positively correlated with the germ cell apoptosis (P＜0. 01 ). Conclusions The decreased expression of SCF and c-kit and increased germ cell apoptosis are found in the contralateral testes, which may contribute to thc infertility in the rat model of experimental unilateral cryptorchidism.     

小儿肾积水尿细胞因子IGF-1、ET-1、TGF-β1及MCP-1水平的检测及意义

Objective To evaluate clinical significance of urinary cytokines in evaluating the damage severity of affected kidneys in children with hydronephrosis. Methods The pyelic urinary in-sulin-like growth factor-1 (IGF-1), endothelin-1 (ET-1), transforming growth factor-β1 (TGF-β1) and monocyte chemoattractant protein-1 (MCP-1) in affected and healthy kidneys (as controls) were de-tected in 41 children with congenital hydronephrosis by enzyme-linked immuno-sorbent assay (ELISA). Pathologic changes of the affected kidneys were graded and the correlations of pathologic grades with urinary level of IGF-1, ET-1, TGF-β1 and MCP-1 were analyzed by Spearman's test. Re-sults The level of urinary IGF-1 in healthy group was (279. 45 ± 31.57) pg/ml, which was signifi-cantly higher than that in hydronephrosis group ( ( 186. 69 ± 24. 63) pg/ml). However, the levels of u-rinary ET-1, TGF-β1 and MCP-1 in healthy group was significantly less than those in hydronephrosis group ((13.21±2.91 vs 49.81 ±9.08) pg/ml, (232.57±32.68 vs 395.91 ±83.52) pg/ml and (328. 54 ± 36. 81 vs 486. 59 ± 89. 72) pg/ml, respectively). Negative correlation was noted between u-rine IGF-1 levels and pathologic grades (P<0. 01 ). Conclusions Urinary IGF-1 is an ideal marker in evaluating the damage severity of affected kidneys in children with hydronephrosis.     

高氧致慢性肺疾病新生大鼠肺泡损伤及肺泡上皮细胞的变化

Objective To explore the changes of alveolar morphology and alveolar epithelial cells in rats with hyperoxia-induced chronic lung diseases (CLD). Methods CLD model in neonatal rats was established by inhalation of high concentration oxygen(85%～90% ). Eighty neonatal rats were randomly exposed to hyperoxia (model group) and to room air (control group) (n =40 each). Radical alveolar counts and the alveolar septum thickness were used to evaluate alveolar development. The site and intensity of expression of SPC,AQP5 protein were detected by immunohistochemical staining,the dynamic expression of SPC mRNA,AQP5mRNA was detected by RT-PCR on day 1,3,7,14 and 21 after exposure. Results There were no significant differences about alveolar wall thickness and RAC between experimental groups and control group on day 1～3 ( P > 0. 05 ). But there was significant difference between the model group and the control groups on day 7 and 14 (P <0. 01 ). For model group,alveolar septum thickness peaked on day 21, the difference was significant compared with control group ( 10. 62±5.01 vs 3.62±0. 88, P < 0. 001 ), but RAC decreased to the lowest level, the difference was significant compared with control group ( 3.57±1.24 vs 10. 47±0. 88,P <0. 001 ). The expression of SPC decreased on day 3 manifestedly but increased on day 7 and the levels of SPC were higher than that in the control group. Experimental group showed gradual decrease in AQP5 expression as the lung impairment devastated. Conclusion Alveolar development was delayed and alveolar epithelial cell (AEC) was damaged in the neonatal CLD rats. The changes of SPC,AQP5 expression suggested AECI was severely damaged and failed in full recovery, meanwhile the quantity of AEC Ⅱ was increased but the ability of its differentiation and transformation was decreased.     

高氧致慢性肺疾病新生大鼠肺泡损伤及肺泡上皮细胞的变化

Objective To explore the changes of alveolar morphology and alveolar epithelial cells in rats with hyperoxia-induced chronic lung diseases (CLD). Methods CLD model in neonatal rats was established by inhalation of high concentration oxygen(85%～90% ). Eighty neonatal rats were randomly exposed to hyperoxia (model group) and to room air (control group) (n =40 each). Radical alveolar counts and the alveolar septum thickness were used to evaluate alveolar development. The site and intensity of expression of SPC,AQP5 protein were detected by immunohistochemical staining,the dynamic expression of SPC mRNA,AQP5mRNA was detected by RT-PCR on day 1,3,7,14 and 21 after exposure. Results There were no significant differences about alveolar wall thickness and RAC between experimental groups and control group on day 1～3 ( P > 0. 05 ). But there was significant difference between the model group and the control groups on day 7 and 14 (P <0. 01 ). For model group,alveolar septum thickness peaked on day 21, the difference was significant compared with control group ( 10. 62±5.01 vs 3.62±0. 88, P < 0. 001 ), but RAC decreased to the lowest level, the difference was significant compared with control group ( 3.57±1.24 vs 10. 47±0. 88,P <0. 001 ). The expression of SPC decreased on day 3 manifestedly but increased on day 7 and the levels of SPC were higher than that in the control group. Experimental group showed gradual decrease in AQP5 expression as the lung impairment devastated. Conclusion Alveolar development was delayed and alveolar epithelial cell (AEC) was damaged in the neonatal CLD rats. The changes of SPC,AQP5 expression suggested AECI was severely damaged and failed in full recovery, meanwhile the quantity of AEC Ⅱ was increased but the ability of its differentiation and transformation was decreased.     

外源性血管内皮生长因子基因对新生鼠缺氧缺血性脑损伤脑细胞凋亡的影响

Objective To explore the effect of exogenesis VEGF 120 gene on the apoptosis of brain cells in the HIBD of newborn rats. Methods VEGF eukaryotic expression plasmid (pCDNA 3.1/r VEGF 120) was constructed by cloning rat VEGF 120 cDNA into eukaryotic expression vector pCDNA 3.1. The HIBD model was established with seven days old SD rats,and all rats were diveded into two groups at random :contral group 18 rats( every rat model was injected pCDNA 3.1 100 μg immediately after hypoxia-is-chemic.then raised seven days) and therapy group 18 rats (every rat model was injected pCDNA 3.1/ rVEGF 120 100 μg immediately after hypoxia-ischemic). Flow cytometer( FCM) was used to detect the ratio of apoptosis of brain cell. Results There was a significant decrease in the ratio of apoptosis brain cells( control group 17.505 ± 0.949; therapy group 8.93 ± 0. 332). Conclusion The VEGF gene product can reduce apoptosis of brain cells.     

外源性血管内皮生长因子基因对新生鼠缺氧缺血性脑损伤脑细胞凋亡的影响

Objective To explore the effect of exogenesis VEGF 120 gene on the apoptosis of brain cells in the HIBD of newborn rats. Methods VEGF eukaryotic expression plasmid (pCDNA 3.1/r VEGF 120) was constructed by cloning rat VEGF 120 cDNA into eukaryotic expression vector pCDNA 3.1. The HIBD model was established with seven days old SD rats,and all rats were diveded into two groups at random :contral group 18 rats( every rat model was injected pCDNA 3.1 100 μg immediately after hypoxia-is-chemic.then raised seven days) and therapy group 18 rats (every rat model was injected pCDNA 3.1/ rVEGF 120 100 μg immediately after hypoxia-ischemic). Flow cytometer( FCM) was used to detect the ratio of apoptosis of brain cell. Results There was a significant decrease in the ratio of apoptosis brain cells( control group 17.505 ± 0.949; therapy group 8.93 ± 0. 332). Conclusion The VEGF gene product can reduce apoptosis of brain cells.     

体位引流辅助气管吸引对胎粪吸入综合征的防治效果

Objective To evaluate the effect of postural drainage assisting trachea suction on meconium aspiration syndrome. Methods Total 61 cases of asphyxia neonates with MAS who were born in our hospital from Jan,2007 to Dec,2008, were divided into control group (24 cases) and observing group( 37 cases). The neonates in control group had endotracheal suction directly after intubation. But the infant in observing group was treated with endotracheal suction after postural drainage. The amount of suction from endotracheal tube,the complication of MAS and the outcome of these newborns were evaluated. Results The total amount of meconium drainage from endotrachea in observing group was statistics significantly more than that in control group [( 2. 16 ± 1.82) ml vs ( 1.23 ± 0. 97 ) ml, P ＜ 0. 05 )]; The intubating times in observing group were statistical significantly less than that in control group[( 1.19 ± 0. 46) vs ( 1.79 ± 0. 83 ) times, P ＜0. 01 ). The incidence of complication in observing group was 8. 11% ,which was significantly lower than that in control group(29. 17% ,P ＜0. 05). There were shorter needing oxygen time [(21.30 ± 22. 38) h vs (52. 91 ±39. 20) h,P ＜0. 01]and shorter hospitalization days [(9. 24 ±3.94) d vs ( 14. 39 ±6. 49) d,P ＜0.01 )]in observing group than those in control group respectively. The mortality in control group was 4. 17%, and no death occurred in observing group. Apgar scores of the first minute was similar in both groups ( P ＞ 0. 05 ). But there was significant difference(70. 16% vs 58. 34% ,respectively;P ＜0. 05) in the fifth minute Apgar scoring of 8 ～ 10 scores between the observing group and the control group. Conclusion Postural drainage assisting endotracheal suction may remove meconium in trachea effectively, decrease the complications of MAS and shorten the oxygen days and hospitalization time.     

白藜芦醇甙对缺氧缺血性脑损伤新生大鼠学习记忆和海马突触素表达的影响

Objective To explore the protective effects and possible mechanisms of Polydatin (PD)on hypoxic-ischemia brain injury(HIBD) in neonatal rat by means of spatial learning memory and the expression of synaptophysin in hippocampal CA1. Methods Thirty-seven neonatal SD rats were divided into 3 groups at random: normal sham-operated group( no hypoxia and ischemia); HIBD group( no medication) ;PD treatment group. 7-old-day rat' s model of HIBD was established by left carotid artery ligation and 2 h hypoxia. Morris water maze test was used to evaluate cognitive function in the rats after 28-day-old( 21-day later after HI). Immunohistochemical method was used to measure the expression of synaptophysin after the end of Morris water maze test. Results Morris water maze results showed that the mean escape latency of the shamgroup (SG) ,HIBD group (HIBD) and PD treatment group (PD) were (39. 55 ±8. 08) s, (52. 37 ±8.03) s and (43.29 ± 7. 63 ) s respectirely. For PD and SG, the mean escape latency was significantly shorter than the HIBD (P ＜0.05). After training,the mean escape latency in the three groups of rats was shortened gradually. The frequency of platform crossings were 5. 29 ±2.62、2. 36 ± 1.80、4. 25 ± 1. 66 in the SG,HIBD and PD respectirely. The frequency of platform crossings in PD was higher than that of HIBD ( P ＜ 0. 05 ). The swimming time in target quadrant were ( 15.74 ± 3.85) s, ( 10. 63 ± 3.66) s and ( 14. 32 ± 2. 52 ) s in SG, HIBD and PD respectirely. For HIBD ,the swimming time in target quadrant was significantly shorter comparing to SG and PD ( P ＜ 0. 05 ). The expression of synaptophys in hippocampal CA1 in PD ( 0. 295 2 ± 0. 044 3 )were evidently higher than that in the HIBD group (0.261 2 ±0.032 3) at 3 week after operation (P ＜0. 05). Conclusion Spatial learning memory deficits and the decrease of synaptophys in hippocampal CA1 could be induced by hypoxic-ischemia. Polydatin could improve the learning and memory ability in neonatal rats following hypoxic-ischemia brain damage. The mechanisms of improvement with Polydatin treatment is associated with the enhancement of expression of synaptophys.     

Specifics of histopathological and genetical diagnosis and classification of lymphomas in children and adolescents

Malignant lymphoma along with leukemias account for nearly half of all malignancies arising in childhood and adolescence. The correct tissue-based histopathological diagnosis of lymphomas results from a close interdisciplinary exchange between pediatric oncologists and hematopathologists. We describe here relevant features of lymphoma subtypes arising in the young age group, Burkitt lymphoma, precursor/lymphoblastic lymphomas, anaplastic large cell lymphoma and diffuse large B-cell lymphoma as well as primary mediastinal B-cell lymphoma and the rare pediatric follicular lymphomas. Special focus is put on specific diagnostic difficulties as well as new insights into biological features of pediatric lymphomas in comparison with their adult counterpart. In addition the relevance of newly defined lymphoma entities of the WHO-classification 2008, e.g. greyzone lymphomas, will be discussed for the young age group.     

Age-dependency of alpha- and beta-adrenoceptors on thrombocytes and lymphocytes of asthmatic and nonasthmatic children

Among the possible mechanisms which may cause wheezing or asthmatic episodes a genetically determined -adrenoceptor blockade and a hyperresponsiveness of -andrenoceptors has been postulated. Evidence to support this hypothesis stems from an increased bronchial sensitivity to -blockers, a reduced formation of cyclic AMP in response to -adrenergic stimulation and enhanced -adrenergic responses in asthmatic subjects. The recent development of techniques for measuring the specific, high-affinity binding of radiolabeled -and -adrenergic antagonists made it possible to study - and -adrenoceptors in vitro. Based upon the assumption that a change in the number and/or affinity of adrenergic receptors might be a general phenomenon, we have performed - and -receptor binding studies on lymphocytes and platelets from wheezing infants and asthmatic children as well as of infants, children, and adults not suffering from these diseases.Using ¹²⁵[I]-cyanopindolol (ICYP) and ³[H]-yohimbine (HYOH) as highly specific ligands for - and -adrenoceptors, the following results were obtained: (1) Lymphocytes and platelets from control subjects and asthamatics bound similar amounts of ICYP and HYOH and thus showed no differences either in the number or the affinity of - and -adrenoceptors. Lymphocytes and platelets of wheezing and nonwheezing infants also bound the same amounts of the radioligands. (2) In asthmatic children receiving 4×2 puffs salbutamol -adrenoceptor were down-regulated and this may mimic -adrenoceptor blockade. (3) When subjects were divided into four categories according to age (0–5, 5–10, 10–20 years, adults) the number of -adrenoceptor binding sites showed an age-dependent increase. The number and affinity of -adreneceptor binding sites on platelets was neither influenced by age nor disease.It is concluded that the - and -adrenoceptors of wheezing infants and asthmatic children at least on blood cells are normal. However the -adrenoceptors show an age-dependent maturation process, which may account for an unresponsiveness to -adrenoceptor agonists in wheezing infants.Supported by a grant from the Ministerium für Wissenschaft und Forschung, NRWPresented at the 19th Workshop for Pediatric Research, University of Göttingen, March 10–11, 1983     

小阴茎、小睾丸的诊断与治疗

小阴茎、小睾丸的病因复杂,临床诊断较为困难。下丘脑-垂体-性腺轴及雄激素合成和转化过程中任何一个环节出现异常,皆会影响阴茎和睾丸的发育。治疗应在明确诊断的基础上予以个体化方案。提倡重视小青春期内的早期诊断,以期早期治疗,使病变的危害降至最低。