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Objective@#To investigate the effect of erythropoietin (EPO) on the expression of aquaporin 2-3 after the release of unilateral ureter obstruction in young rats.@*Methods@#Twenty-four SD rats were randomly divided into 3 groups(CUUO-R group, CUUO-R+ EPO group and sham group, with 8 rats in each group). The CUUO-R model was built through unilateral ureteral ligation, after 48 h the obstruction was released.EPO was given to the CUUO-R+ EPO group at the time point of removing obstruction, and then repeated every other day for 1 week, and the same volume of saline was simultaneously given to the CUUO-R rats.The rats in sham group experienced the laparotomy and free dissection of left ureter but not ligation.The kidneys were harvested 7 d after the release of CUUO.The methods of Western blot and immunohistochemistry were used to examine the effects of erythropoietin on the expression of AQP2 and AQP3.@*Results@#The osmotic pressure of CUUO-R+ EPO group was higher than those of CUUO-R group, but lower than that of sham group(P=0.007). The concentration of creatinine and urea in the CUUO-R group[(58.001±2.416) μmol/L and (9.025±1.158) mmol/L]were higher than those of CUUO-R+ EPO group [(57.072±2.286) μmol/L and (1.479±0.043) mmol/L] and sham group [(54.820±1.536) μmol/L and (6.929±0.604) mmol/L]. The differences of the concentration of creatinine and urea between CUUO-R group and sham group were statistically significant(P<0.05). There was no significant difference between CUUO-R+ EPO group and Sham group(P>0.05). The immunohistochemistry showed that the expressions of AQP2 and AQP3 in co-llecting duct in CUUO-R group were significantly weaker than those of in sham group, and the expression of those in CUUO-R+ EPO group were slightly weaker than sham group.These results were further confirmed by Western blot, as the relative quantity of AQP2 and AQP3 were also the lowest in CUUO-R group(AQP2 in 3 groups were 0.974±0.109, 1.923±0.097 and 2.002±0.044, F=392.4, P=0.000; AQP3 in 3 groups were 0.941±0.048, 1.497±0.043 and 1.863±0.043, F=735.8, P=0.000).@*Conclusions@#EPO treatment is beneficial for the recovery expre-ssion of AQP2 and AQP3 as well as renal function at the early period after the release of ureteral obstruction in young rats.
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Objective To investigate the effect of erythropoietin(EDO)on the expression and function of aqua_porin_1( AQD _1)in the kidney of young male SD rats after release of bilateral ureter obstruction( BUO _ R). Methods Porty_eight young SD rats were randomly divided into bilateral ureteral complete obstruction(BUO)group (n﹦6),BUO_R group(n﹦12),BUO_R﹢EDO group( n﹦12)and Sham group( n﹦18). The BUO model was built through bilateral ureteral ligation. BUO group were killed after 24 h,and BUO_R group and BUO_R﹢EDO group were relieved after obstruction of 24 h. EDO(500 U∕kg)was given to BUO_R﹢EDO rats at 1 h after release of BUO,and then repeated 1 d,3 d and 5 d thereafter and the same volume of 9 g∕L saline was simultaneously given to BUO_R rats. The Sham group was prepared in parallel by laparotomy and free dissection of bilateral ureters but not ligated,both side kidneys and blood samples were collected on 3 d and 7 d(24 h,3 d,7 d for Sham group)after release of BUO. The u_rine samples were collected by using metabolic cage before death. The plasma osmotic pressure,creatinine(Cr)and u_rea nitrogen(BUN)in the plasma of young rats were detected. The expression of AQD_1 protein in all groups of kidney tissues was detected by adopting immunohistochemistry and Western blot. Results On day 3 after release of BUO,24 h water intake and urine volume of BUO_R﹢EDO group were higher than those of Sham group,but lower than those of BUO group(P〈0. 05),the urine osmotic pressure of BUO_R﹢EDO group was higher than that of BUO group,but lower than that of Sham group(P〈0. 05),while plasma osmotic pressure,Cr and BUN of BUO_R﹢EDO group were higher than those of Sham group,but lower than those of BUO group(P〈0. 05),and they were all of lower than BUO group( P 〈0. 05). On day 7 after release of BUO,there was no obvious change in Sham group,and the indexes of BUO_R group and BUO_R﹢EDO group gradually recovered,but they still did not reach the normal level(P〈0. 05). The difference between BUO_R group and BUO_R﹢EDO group was statistically significant(P〈0. 05). The immuno_histochemical results showed that the expression of AQD_1 in collecting duct in BUO group was significantly down_regulated compared with that in Sham group,whereas it was slightly weaker in BUO_R group and BUO_R﹢EDO group than that of Sham group(P〈0. 05). Compared with 3 days after release of BUO,the staining intensity of BUO_R﹢EDO group and BUO_R group was enhanced,but still lower than that of the Sham group. These results were further confirmed by adopting Western blot,and BUO group was also the lowest of the four groups,and BUO_R﹢EDO group was higher than that of BUO group,but lower than that of Sham group( P〈0. 05). Conclusion EDO can promote not only the recovery of AQD_1 protein expression but also the recovery of renal function in young BUO_R rats.
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Objective To investigate the effect of erythropoietin (EPO) on the expression of aquaporin 2-3 after the release of unilateral ureter obstruction in young rats.Methods Twenty-four SD rats were randomly divided into 3 groups(CUUO-R group,CUUO-R + EPO group and sham group,with 8 rats in each group).The CUUO-R model was built through unilateral ureteral ligation,after 48 h the obstruction was released.EPO was given to the CUUO-R + EPO group at the time point of removing obstruction,and then repeated every other day for 1 week,and the same volume of saline was simultaneously given to the CUUO-R rats.The rats in sham group experienced the laparotomy and free dissection of left ureter but not ligation.The kidneys were harvested 7 d after the release of CUUO.The methods of Western blot and immunohistochemistry were used to examine the effects of erythropoietin on the expression of AQP2 and AQP3.Results The osmotic pressure of CUUO-R + EPO group was higher than those of CUUO-R group,but lower than that of sham group (P =0.007).The concentration of creatinine and urea in the CUUO-R group [(58.001 ± 2.416) μmol/L and (9.025 ± 1.158) mmol/L] were higher than those of CUUO-R + EPO group [(57.072 ± 2.286) μmol/L and (1.479 ± 0.043) mmol/L] and sham group [(54.820 ± 1.536) μmol/L and (6.929-±0.604) mmol/L].The differences of the concentration of creatinine and urea between CUUO-R group and sham group were statistically significant (P ＜ 0.05).There was no significant difference between CUUO-R + EPO group and Sham group(P ＞ 0.05).The immunohistochemistry showed that the expressions of AQP2 and AQP3 in collecting duct in CUUO-R group were significantly weaker than those of in sham group,and the expression of those in CUUO-R + EPO group were slightly weaker than sham group.These results were further confirmed by Western blot,as the relative quantity of AQP2 and AQP3 were also the lowest in CUUO-R group (AQP2 in 3 groups were 0.974 ± 0.109,1.923 ± 0.097 and 2.002 ± 0.044,F =392.4,P =0.000;AQP3 in 3 groups were 0.941 ± 0.048,1.497 ± 0.043 and 1.863 ± 0.043,F =735.8,P =0.000).Conclusions EPO treatment is beneficial for the recovery expression of AQP2 and AQP3 as well as renal function at the early period after the release of ureteral obstruction in young rats.
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Objective To investigate the value of detrusor isovolumetric test(DIT)on female SUI with sus-pected detrusor contraction weakness. Methods A total of 86 SUI patients with suspected detrusor contraction weakness diagnosed by preoperative bladder cystometry(CMG)were enrolled in this study.DIT method was used to evaluate the bladder function. The difference of first sensation cystometry capacity, maximum cystometric capacity, bladder compliance and maximum detrusor pressure were compared between the two methods.Patients in group Ⅰwhowere diagnosed with normal detrusor function with DIT method underwent TOT surgery. Ninety normal SUI patients were selected as control group Ⅱ,who were diagnosed with normal detrusor function with CMG method. The surgery effect was compared between the two groups. Results By using DIT detection,there were 65 cases showed normal detrusor function of the 86 cases(75.6%).The maximum detrusor pressure with DIT was(31.2 ± 12.9)cm H2O versus(11.7 ± 3.1)cm H2O with cystometry(P＜0.05).No significant differences were observed in the first sensation cystometry capacity,maximum cystometric capacity and bladder compliance.The effective rate of TOT in both group Ⅰ and group Ⅱ was more than 95%,with no significantly difference between the two groups (P＞0.05). Conclusions DIT is a valuable method to evaluate the bladder function of female SUI patients with suspected detrusor contraction weakness.
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Objective To explore the expression of aquaporin-2 (AQP-2) in human fetus kidney and amniotic fluid at different stages of pregnancy.Methods Twenty-two cases of aborted fetuses' kidneys were collected.They were divided into 3 groups according to the pregnancy age:8 cases in 17-23 + 6 weeks,8 cases in 24-31 +6 weeks,and 6 cases in 32-38 +6 weeks.Western blot was used to examine the expression of AQP-2 in the kidney.Twenty-four cases of the amniotic fluid were collected,and they were divided into 3 groups according to the pregnancy age:10 cases in 17-23 +6 weeks,6 cases in 24-31 +6 weeks,and 8 cases in 32-38 +6 weeks.Eight cases of healthy adult morning urine were collected as positive controls.The AQP-2 protein in the amniotic fluid was detected with the method of enzyme-linked immunosorbent assay (ELISA) and the osmotic pressure of amniotic fluid at different stages of the pregnancy was measured with the freezing point osmometer.Results The expression of AQP-2 was increased with the extending of pregnancy age,and the AQP-2 expressions in fetus kidney of 17-23 +6 weeks,24-31 + 6 weeks and 32-38 +6 weeks were 0.986 ± 0.335,1.566 ± 0.272,and 2.080 ± 0.246,respectively,and the difference was significant (P ＜ 0.05).The AQP-2 detected from amniotic fluid was positively correlated with the result of AQP-2 in the kidney(r =0.985,P ＜ 0.05),and the AQP-2 expression also increased with the extending of pregnancy age:17-23 +6 weeks,24-31 +6 weeks,32-38 +6 weeks and adult urine was (30.253 ±5.843) mg/L,(35.103 ±7.271) mg/L,and (42.580 ± 1.230) mg/L and (46.493 ± 0.450) mg/L,respectively.The osmolality of the amniotic fluid of 17-23 +6 weeks,24-31 +6 weeks,32-38 +6 weeks was (272.600 ± 4.827) mmol/L,(252.00 ± 15.360) mmol/L,and (261.750 ±5.560) mmol/L,respectively,and the difference was significant(P ＜0.05).Conclusions The AQP-2 expression in human fetus kidneys has good correlation with amniotic fluid,which indicates that the level of AQP-2 of the amniotic fluid may reflect the expression of AQP-2 in the fetus kidney.
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Objective To investigate the effects of Forskolin on the activation of nucleotide－binding oligome-rization domain like receptor family,pyrin domain－containing 3(NLRP－3)inflammasome and the secretion of inter-leukin－1β(IL－1β)in activated human THP－1 macrophages,which can provide evidence for clinical treatment of inflammatory diseases in children.Methods Human monocyte cell line THP－1 cells were induced to differentiate into macrophages by Phorbol－12－myristate－13－acetate(PMA),and Forskolin(10,50,100 μmol/L)stimulated activa-ted macrophages by nigericin.The mRNA of the inflammasome markers NLRP－3,IL－1β and Caspase－1 were detec-ted by real－time quantitative polymerase chain reaction(qPCR)method.The protein of NLRP －3,pro －IL －1β, pro－Caspase－1 and Caspase－1 were detected by Western blot.The secretion of inflammatory cytokines IL－1β was detected by enzyme－linked immuno－sorbent assay(ELISA).Results Nigericin activated the cells and the mRNA expression of NLRP－3,IL－1β and Caspase－1 increased by 7.59 times(P＜0.001),579.10 times(P＜0.001)and 3. 59 times (P ＜0. 001 ),compared to non － activated cells;Forskolin had no effect on the mRNA expression of NLRP－3,Caspase－1 and IL－1β on activated THP－1 macrophages(P＞0.05).Western blot showed that Forskolin also had no effect on the protein expression of pro－Caspase－1 and pro－IL－1β in activated THP－1 macrophages (P＞0. 05),but the expression of NLRP －3 and Caspase －1 decreased significantly (P ＜0.001). The results of ELISA showed that the IL －1β secretion increased from basal 584. 0 nmol/L to activated 2 695. 6 nmol/L (t ＝16.031 1,P＜0. 001)on THP －1 macrophages by nigericin,but Forskolin(10,50,100 μmol/L)reduced it to 1 858. 4 nmol/L(t＝5.365 5,P ＜0. 001),1 467.9 nmol/L(t ＝8.047 5,P ＜0.001)and 1 246.7 nmol/L(t ＝10.199 0,P＜0.001)on the activated THP－1 macrophages.Conclusions Forskolin can not affect the expression of pro－cytokines at the gene transcription and protein translation levels in activated THP－1 cells,but inhibit the secre-tion of inflammatory cytokines IL－1β,so as to inhibit inflammatory response,which can be used to treat pediatric in-flammatory diseases caused by IL－1β.
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Objective To investigate the effect of early application of clean intermittent catheterization(CIC) in infants with neurogenic bladder(NB).Methods Eighty-seven children with NB diagnosed in our urodynamic center were less than 1 year old when they first came to hospital from January 2007 to January 2010, and CIC was carried out at different age.Sixty-four patients were followed up for a long time and divided into early CIC group(less than 1 year old children) and late CIC group(more than 3 years old children) according to the treatment time.Early CIC group included 29 patients [19 boys and 10 girls with the mean age of (7.5 ±2.8) months].And 4 cases were suffering from postoperative spina bifida manifesta;22 cases with spina bifida occulta;2 cases with sacral dysplasia;1 case with meningitis.Late CIC group included 35 patients [20 boys and 15 girls with the mean age of (8.0 ±2.9) months].2 cases were suffering from postoperative spina bifida manifesta;28 cases with spina bifida occulta;4 cases with sacral dysplasia;1 case with postoperative pelvic surgery.Before the treatment, there were no significant differences of the bladder compliance (BC), the maximum cystometric capacity (MCC) and the safety bladder capacity (SBC) between two groups.Urodynamic parameters and complications of 64 patients who were successfully followed up for 6 years were compared.Results After 3 years follow up, BC, SBC and MCC in early CIC group [(8.5 ± 1.9) ml/cmH2O, (140 ±25) ml, (142 ±29) ml]were significantly higher than those of late CIC group [(7.0 ± 2.2) ml/cmH2O, (110 ± 31) ml, (120 ± 28) ml;all P ＜ 0.05].After 6 years follow up, BC, SBC and MCC in early CIC group [(12.0 ±2.5) ml/cmH2O, (210 ±26) ml, (230 ±30) ml] were significantly higher than those of late CIC group [(9.3 ± 2.3) ml/cmH2O, (192 ± 31) ml, (205 ± 35) ml;all P ＜ 0.05], and the vesicoureteral reflux rate [24.1％ (7/29)] in early treatment group was significantly less than that in late treatment group [54.3％ (19/35), P ＜ 0.05].Increases in BUN and serum creatinine were found in 6 cases (20.7％) in early CIC group and 17 cases (48.6％) in late CIC group, the difference was significant (P ＜ 0.05).Conclusion For NB patients, the effect of early CIC is better than that of late CIC.
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Objective To explore the expressions of histamine receptor subtypes (H1, H2, H3, H4 receptor) in children's mid-urethral striated muscles and during mouse C2C12 striated myogenesis.Methods Children's mid-urethral striated muscle samples were paraffin embedded and tissue sections were made, then immunohistochemical staining was used to check H1, H2, H3, H4 receptors.C2C12 myogenesis was induced, the early differentiation early markers of desmin, middle marker of myogenin, late marker of myosin heavy chain and histamine 4 receptor subtype mRNA were checked by quantitative real-time polymerase chain reaction.Immunofluorescence staining was done to check 3 differentiation markers and histamine H3 receptor protein.Results During myogenesis, the expression of desmin mRNA in the differentiation of 2,4,6 days were 12,68,60 times as many as that of the undifferentiated myoblasts;the expression of myogenin mRNA in the differentiation of 2,4,6 days were 631,1 408,914 times as many as that of the undifferentiated myoblasts;the expression of myosin heavy chain mRNA in the differentiation of 2,4,6 days were 7 718,9 448,286 288 times as many as that of the undifferentiated myoblasts.The expression level of H1 receptor mRNA in the differentiation of 6 days was about 25％ to undifferentiated cells;the expression of H2 receptor mRNA in undifferentiated cells and differentiated cells groups had no significant difference (F =1.47, P ＞ 0.05);the expression of H3 receptor mRNA in the differentiation of 2,4,6 days was 28,103,198 times to undifferentiated cells;H4 receptor mRNA was not detected.In immunofluorescence staining, H3 receptor protein staining intensity increased with the differentiation.Immunohistochemistry of pediatric urethral striated staining indicated that H1, H2, H3 receptor staining was positive,H1 receptor showed strong positive staining, H3 receptor moderate positive staining,and H2 receptor showed weak positive staining.Conclusions Histamine receptor subtypes of H1 receptor, H2 receptor and H3 receptor were found during mouse striated myogenesis and in the children's mid-urethral striated muscles.The increasing expression of H3R with myogenesis might indicate it plays a role in mature striated muscle cells.