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<p><b>OBJECTIVE</b>To investigate the effects of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) of TGF-β1-induced human renal proximal tubular epithelial (HK-2) cells.</p><p><b>METHODS</b>The gene sequence of miR-145 was synthesized and cloned into pCMV-myc to construct recombinant plasmid pCMV-miR-145. HK-2 cells were divided into four groups: control (untreated), TGF-β1 (treated with TGF-β1), blank+TGF-β1 (treated with TGF-β1 after HK-2 cells transfected with blank plasmid) and miR-145+TGF-β1 (treated with TGF-β1 after HK-2 cells transfected with pCMV-miR-145 recombinant plasmid). Expression of miR-145 was detected by real-time PCR (RT-PCR). TGF-β1, Smad3, Smad2/3, p-Smad2/3, α-SMA, FN and type I collagen (Col I) protein levels were detected by Western blot. Concentrations of fibronectin (FN) and Col I in cell culture supernatants were measured using ELISA.</p><p><b>RESULTS</b>pCMV-miR-145 recombinant plasmid was successfully transfected into HK-2 cells. Compared with the control group, the miR-145+TGF-β1 group showed a significant up-regulation in the expression level of miR-145 (P<0.01). However, the TGF-β1 and blank+TGF-β1 groups showed a significant down-regulation in the expression level of miR-145 compared with that in the control and miR-145+TGF-β1 groups (P<0.01). Compared with the TGF-β1 and blank+TGF-β1 groups, the miR-145+TGF-β1 group showed significantly reduced levels of the signal proteins TGF-β1, Smad3, Smad2/3 and p-Smad2/3 (P<0.05), as well as significantly reduced levels of the biomarkers α-SMA, FN and Col I (P<0.05). Meanwhile, concentrations of FN and Col I in cell culture supernatants also decreased (P<0.05).</p><p><b>CONCLUSIONS</b>miR-145 modulates the EMT of HK-2 cells treated with TGF-β1, possibly by inhibition of the activation of TGF-β-dependent Smad signaling pathway.</p>
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Humanos , Células Cultivadas , Células Epiteliais , Patologia , Transição Epitelial-Mesenquimal , Túbulos Renais Proximais , Patologia , MicroRNAs , Fisiologia , Fator de Crescimento Transformador beta1 , FarmacologiaRESUMO
OBJECTIVE@#To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD).@*METHODS@#The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05).@*CONCLUSIONS@#As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.
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Objective To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD). Methods The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA). Results The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05). Conclusions As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.
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Objective To analyze the correlation between clinical characteristics and pathological findings lupus nephritis (LN) in children,together with the correlation of the renal vascular lesion respectively with the glomerular lesion and tubulointerstitial lesion.Methods Forty-one cases of LN in children diagnosed by percutaneous renal biopsy from Jan.2008 to Sep.2012 in Pediatrics of the Second Xiangya Hospital of Central South University were collected.Clinical manifestations and the lab findings of the blood and urine of all the patients were analyzed,and all the frozen sections were evaluated according to the standard of ISN/RPS2003 for LN.The glomerular lesion and tubalointerstitial lesion separately were also evaluated,respectively.The wall thickening/outer diameter ratio,intima thickening/outer diameter ratio and medial thickening/outer diameter ratio of the arterioles were measured.Results The percentage of clinical manifestation with LN increased coupled with the degree of pathological damage.In the different stages of pathological damage,both the glomerular lesion and tubulointerstitial lesion were getting much more serious along with the progressiveness of pathological damage,and what's more,they had a positive correlation(r =0.959,P < 0.05).The wall thickness/outer diameter ratio in all cases was greater than 0.5,in dicating the thickness of vessel wall.Conclusion Renal vascular lesion really existed and is characterized by the progressive loss of integrity of the intima and the medial thickening.
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Objective To research the changes of clinical symptoms and serum free radical in children with severe Henoch-Sch(o)nlein purpura (HSP) between before and after accepted hemoperfusion.And evaluate the curative effect of hemoperfusion treatment for severe HSP and discuss the mechanism.Methods Twenty-three severe HSP patients in Children's Medical Center of the Second Xiangya Hospital of Central South University which were divided into 2 groups:13 cases were divided into traditional treatment group,10 cases were divided into hemoperfusion group; 11 healthy children were divided into healthy control group.The alleviate situation of clinical symptoms were observed and recorded such as purpura,abdominal pain,joint pain,hematuria,albuminuria,and the changes of urine RBC count,24 hour urine protein quantitative before and after hemoperfusion and traditional treatment.Collected the serum before and after the first time hemoperfusion treatment,after the second time hemoperfusion treatment,after the third time hemoperfusion treatment in hemoperfusion group;Collected the serum before and after conventional therapy in traditional treatment group; Collected only once serum in healthy control group.And then their superoxide anion(O2-·),hydroxy radical(· OH),hydrogen dioxide(H2O2),malonaldehyde(MDA) values were detected with spectrophotometry.The differences of each index among each group were compared respectively.Results After treatment,the clinical symptoms of 23 severe HSP children such as rash,abdominal pain,joint pain,hematuria,albuminuria which were reliefed compared to before treatment.The urine erythrocyte count,24 hour urine protein quantitative were reduced in 2 groups,but the symptoms of perfusion group children relieved faster,and the clinical index decreased more obviously.The serum O2-·,· OH,H2O2,MDA levels of 23 HSP children were significantly higher than those of the healthy control group (all P < 0.01).In the traditional treatment group,compared between before and after treatment,the indexes were decreased (all P < 0.05) ; After 3 times of hemoperfusion,all indexes were decreased in hemoperfusion group,but only after the third hemoperfusion the indexes were decreased statistically significant(all P < 0.05).But compared with the hemoperfusion group,the index were decreased more apparently in after the third hemoperfusion,and it was statistically significant (P < 0.05).Conclusions 1.The serum free radicals are increased in severe HSP children,they may play a role in vasculitis.2.For severe HSP,the recent therapeutic effect of hemoperfusion ally with traditional treatment is better than the alone traditional treatment.3.Hemoperfusion ally with traditional treatment can remove more effectly the serum free radicals,reduce lipid peroxide products,then mitigate the damage of the oxidate stress to the vascular endothelial.
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<p><b>OBJECTIVE</b>To explore possible correlations between renal Th1/Th2 ratio and renal microvascular injury in children with Henoch-Sch-nlein purpura nephritis (HSPN).</p><p><b>METHODS</b>Thirty-two children with HSPN were enrolled. They were classified into four groups by renal pathology: HSPN class II (n=8), HSPN class IIIa (n=7), HSPN class IIIb (n=10) and HSPN class IV/V (n=7). Five patients undergoing nephrectomy due to trauma were used as the controls. INFγ, IL-4 and CD34 in the renal tissues were measured by immunohistochemical analysis. INFγ was used as a marker of Th1, IL-4 was used as a marker of Th2 and CD34 was used as a marker of microvessel. The renal microvessel density was evaluated according to the Weidner standard. The relationships among the local Th1/Th2 ratio, renal pathological grade, microvessel score and microvessel density were studied.</p><p><b>RESULTS</b>Immunohistochemical analysis showed a lower expression of INFγ and a higher expression of IL-4 in the HSPN groups than in the control group. The local Th1/Th2 ratio in the HSPN groups decreased and correlated significantly with the renal pathological grade. There were significant differences among four HSPN subgroups (P<0.05). Compared with the control group, the renal microvessel density in the HSPN class II and class IIIa groups increased significantly (P<0.05), but it decreased in the HSPN class IV/V group (P<0.05). The renal microvessel scores in the HSPN class IIIa, class IIIb and class IV/V groups increased significantly compared with those in the control and the HSPN classⅡ. The increased renal microvessel scores were associated with more severe renal pathological changes. A negative correlation was found between the local Th1/Th2 ratio and the microvessel density in kidneys (r=-0.921, P<0.01).</p><p><b>CONCLUSIONS</b>The decrease of Th1/Th2 ratio in kidneys might be responsible for renal microvascular injury in children with HSPN.</p>
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Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Rim , Patologia , Microvasos , Patologia , Nefrite , Alergia e Imunologia , Patologia , Vasculite por IgA , Alergia e Imunologia , Patologia , Células Th1 , Alergia e Imunologia , Células Th2 , Alergia e ImunologiaRESUMO
<p><b>OBJECTIVE</b>To study possible influences of 1,25(OH)(2)D(3) on endothelial cell proliferation, apoptosis and endothelial nitric oxide synthase (eNOS) expression of aorta in apolipoprotein E-deficient (apoE(-/-)) mice and to explore the relationship between vitamin D and atherosclerosis.</p><p><b>METHOD</b>Endothelial cell of aorta in apoE(-/-) mice were isolated and cultured, and the influence of 1,25(OH)(2)D(3) on endothelial cell proliferation were observed by MTT, apoptosis of cells were quantitated by terminal deoxynucleotidyl transferase mediated dUTP nick end labelling, Bcl-2 mRNA, fas mRNA and eNOS mRNA was detected by reverse transcription-polymerase chain reaction.</p><p><b>RESULT</b>Endothelial cell proliferation rate of aorta did not significantly change in the two control groups (0.162 ± 0.031 vs. 0.158 ± 0.006, P > 0.05). Compared with control groups, 1,25(OH)(2)D(3) stimulated endothelial cell proliferation of aorta (P < 0.05), but endothelial cell proliferation rate did not significantly change in different 1,25(OH)(2)D(3) concentration groups [1,25(OH)(2)D(3) concentration: 10(-4)mol/L, 10(-5) mol/L, 10(-6) mol/L, 10(-7) mol/L, 10(-8) mol/L, endothelial cell proliferation rate: 0.189 ± 0.013 vs. 0.285 ± 0.011 vs. 0.296 ± 0.026 vs. 0.284 ± 0.017 vs. 0.233 ± 0.010, P > 0.05]. 1,25(OH)(2)D(3) research concentration as chosen as 10(-6) mol/L. In 1,25(OH)(2)D(3) 10(-6) mol/L group, the expression of Bcl-2, eNOS mRNA was significantly increased (0.78 ± 0.16 vs. 0.46 ± 0.21 vs. 0.42 ± 0.17, 0.56 ± 0.16 vs. 0.39 ± 0.13 vs. 0.35 ± 0.11, 0.46 ± 0.2 vs. 10.42 ± 0.17 vs. 0.78 ± 0.16, 0.79 ± 0.21 vs. 0.81 ± 0.20 vs. 0.43 ± 0.12), apoptotic index, Fas mRNA was significantly decreased (15.14 ± 3.19 vs. 18.94 ± 4.22 vs. 19.27 ± 4.58, 0.43 ± 0.12 vs.0.79 ± 0.21 vs. 0.81 ± 0.20)(P < 0.05). The quantity of eNOS gene expression was inversely associated with apoptosis index and Fas mRNA, was positively associated with Bcl-2 mRNA (r = -0.676, -0.758, 0.762, P < 0.01).</p><p><b>CONCLUSION</b>1,25(OH)(2)D(3) stimulated endothelial cell proliferation, inhibited apoptosis and increased eNOS expression of aorta in apoE(-/-) mice. These results may deepen understanding of the pathogenesis of atherosclerosis.</p>
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Animais , Feminino , Masculino , Camundongos , Aorta , Metabolismo , Apolipoproteínas E , Apoptose , Calcitriol , Farmacologia , Proliferação de Células , Células Cultivadas , Células Endoteliais , Metabolismo , Óxido Nítrico Sintase Tipo III , Metabolismo , RNA Mensageiro , GenéticaRESUMO
<p><b>OBJECTIVE</b>To investigate the changes of blood pressure by 24-hour ambulatory blood pressure (ABP) monitoring in children with primary nephrotic syndrome (PNS) and explore the relationship of the changes in blood pressure with rennin-angiotensin-aldosterone system (RAAS) in these children.</p><p><b>METHODS</b>ABP and casual blood pressure (CBP) monitoring were performed in 114 children with PNS. Plasma levels of rennin activity (PRA), angiotensin II (AngII) and aldosterone (ALD) were measured. The correlation of plasma levels of PRA, AngII and ALD with ABP was evaluated.</p><p><b>RESULTS</b>Of the 114 children with PNS, 101 (88.6%) presented elevated blood pressure. Mild or severe masked hypertension was found in 45 children (39.5%). Eighty (70.2%) children showed non-dipper blood pressure. The index and load of systolic blood pressure were higher than those of diastolic blood pressure. The blood pressure index and blood pressure load during sleep were higher than those during wakefulness. The boy presented higher diastolic blood pressure index and load than girls. Decubitus blood PRA, AngII and ALD levels in children with PNS were significantly higher than normal controls. The group with elevated blood pressure presented significantly higher decubitus blood PRA, AngII and ALD levels than the group with normal blood pressure. AngII level was significantly positively correlated with the index and load of both systolic blood pressure and diastolic blood pressure.</p><p><b>CONCLUSIONS</b>The children with PNS present a high incidence of hypertension, with a large percentage of masked hypertension and non-dipper blood pressure. Systolic blood pressure increases more significantly than diastolic blood pressure. Blood pressure during sleep increases more significantly than that during wakefulness. Diastolic blood pressure increases more significantly in boys than in girls. RAAS activity is elevated and the elevated RAAS activity might increase the blood pressure mainly by AngII in children with PNS.</p>
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Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pressão Sanguínea , Monitorização Ambulatorial da Pressão Arterial , Síndrome Nefrótica , Sistema Renina-Angiotensina , FisiologiaRESUMO
<p><b>OBJECTIVE</b>To study the effect of H2O2 on the proliferation and apoptosis of endothelial progenitor cells (EPCs) and the antogonistic effects of catechin on the cell apoptosis induced by H2O2 in rats.</p><p><b>METHODS</b>Immuno-fluoreascence assay was applied to detect CD34, CD133 and VEGFR-2 expression. EPCs of generation 2 were divided into control cells, H2O2-treated cells and catechin-H2O2-treated cells (H2O2: 100 mg/L; catechin: 10 mg/L). Genomic DNA was extracted by the conventional method after intervention for the analysis of apoptosis ladder pattern. The MTT assay was applied to detect proliferation rate of EPCs.</p><p><b>RESULTS</b>The cultured cells at day 10 expressed CD34, CD133 and VEGFR-2. DNA apoptosis ladder pattern appeared in H2O2-treated cells 2 days after intervention. After 3 days of intervention DNA apoptosis ladder pattern appeared in both H2O2-treated cells and H2O2-catechinjtreated cells, with more ladders and grayer scale in H2O2 -treated cells. Compared with the controls, H2O2-treated cells and H2O2-catechin-treated cells showed significantly decreased proliferation rate (p<0.01), with the lowest proliferation rate at the 2nd day (p<0.05). The H2O2-catechin-treated cells showed increased proliferation rate than H2O2-treated cells at the 1st, 2nd and 3rd days.</p><p><b>CONCLUSIONS</b>H2O2 may impair EPCs proliferation and induce EPCs apoptosis. Catechin may increase the capacity of EPCs for the resistance to apoptosis induced by H2O2.</p>
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Animais , Feminino , Ratos , Antígeno AC133 , Antígenos CD , Antígenos CD34 , Apoptose , Catequina , Farmacologia , Proliferação de Células , Células Endoteliais , Biologia Celular , Glicoproteínas , Peróxido de Hidrogênio , Toxicidade , Peptídeos , Ratos Sprague-Dawley , Células-Tronco , Biologia Celular , Receptor 2 de Fatores de Crescimento do Endotélio VascularRESUMO
<p><b>OBJECTIVE</b>To investigate the relationship between vascular endothelial growth factor (VEGF) expression and microvessel injury of renal interstitium in children with Henoch-Schönlein purpura nephritis (HSPN).</p><p><b>METHODS</b>Thirty-two children with HSPN and who had not received glucocorticoid or immunodepressants treatment before hospitalization were enrolled. Five children undergoing nephrectomy due to renal trauma were used as the control group. Renal samples were stained by hematoxylin and eosin and renal pathological changes were evaluated semi-quantitatively. CD34 and VEGF expression was detected by immunohistochemistry. CD34 was used as the marker for endothelial cells of renal microvessels. The microvessel density (MVD) was calculated by CD34 immunostaining.</p><p><b>RESULTS</b>Compared with the control and the renal pathological grade II HSPN groups, MVD in the grade III and above HSPN groups decreased significantly, with an obvious reduction in MVD with the increased renal pathological grade (p<0.05). The renal microvessel score in the grades IIIa, IIIb, IV, and V HSPN groups decreased obviously compared with that in the control group. The renal microvessel score decreased with the increased renal pathological grade (p<0.05). VEGF expression in the grade II HSPN group was higher (p<0.05), while that in the grades IV and V HSPN group was lower than that in the control group (p<0.05). VEGF expression in the HSPN group showed a significant reduction with the increased renal pathological grade (p<0.05). There was a positive correlation between VEGF expression and MVD in renal tissue in the HSPN group (r=0.935, p<0.01).</p><p><b>CONCLUSIONS</b>The decreased expression of VEGF may be responsible for the renal pathological damage and microvessel injury in HSPN.</p>
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Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Imuno-Histoquímica , Rim , Química , Microvasos , Patologia , Nefrite , Vasculite por IgA , Metabolismo , Patologia , Fator A de Crescimento do Endotélio VascularRESUMO
<p><b>OBJECTIVE</b>To evaluate the effect of clearance of superoxide anion by catechin on the expression of nitrogen monoxidum (NO) and endothelial nitricoxide synthase (eNOS) and apoptosis in endothelial progenitor cells (EPCs) induced by angiotensin II (Ang II).</p><p><b>METHODS</b>The marrow endothelial progenitor cells of Sprague-Dawley rats were isolated and assigned to control (no treatment), Ang II treatment and Ang II + catechin treatment groups. After 48 hrs of culture, the concentration of O2*- in the supernate was measured by the NBT method, and NO concentration in the supernate was measured by the nitrate reductase method; the apoptosis rate of EPCs was detected by the TUNEL method; the mRNA expression of eNOS was detected by RT-PCR; the protein expression of eNOS was detected by Western blot analysis.</p><p><b>RESULTS</b>Ang II of 10-6 mol/L was determined as the suitable concentration for cell induction by the MTT test. Catechin of 400 mg/L was determined as an advisable intervention dosage. The apoptosis rate of EPCs in the control, the Ang II and the Ang II+catechin treatment groups were 2.48+/-0.12%, 54.18+/-0.77% and 16.87+/-0.35%, respectively, and there were significant differences among the three groups (P<0.01). The O2*- concentration in the Ang II and the Ang II+catechin treatment groups (81.7+/- 3.6 and 62.3+/- 2.2 U/L respectively) was significantly higher than that in the control group (33.7+/- 2.8 U/L) (P<0.01). An increased NO concentration was also found in the Ang II (189. 8+/- 9.0 micromol/L) and the Ang II+catechin treatment groups (276.4+/- 10.1 micromol/L) compared with that in the control group (105.8+/- 9.8 micromol/L) (P<0.01). There were significant differences in the concentrations of O2*- and NO between the Ang II and the Ang II+catechin treatment groups (P<0.05). The mRNA (P<0.05) and protein expression (P<0.01) of eNOS in the Ang II and the Ang II+catechin treatment groups increased significantly compared with those in the control group. The Ang II+catechin treatment group showed increased eNOS protein expression compared with the Ang II group (P<0.05).</p><p><b>CONCLUSIONS</b>Ang II may induce the generation of O2*-, inactivate NO and increase gene and protein expression of eNOS in EPCs. Catechin might decrease the apoptosis of EPCs through the effective clearance of O2*-and the reduction of NO inactivation and of eNOS protein uncoupling.</p>
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Animais , Feminino , Ratos , Angiotensina II , Farmacologia , Apoptose , Catequina , Farmacologia , Sobrevivência Celular , Células Endoteliais , Metabolismo , Óxido Nítrico , Óxido Nítrico Sintase Tipo III , Genética , RNA Mensageiro , Ratos Sprague-Dawley , Células-Tronco , Metabolismo , Superóxidos , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To study the regulattory effect of Astragalus membranaceus on immune disturbance of the rats with IgA nephropathy.</p><p><b>METHODS</b>Rats IgA nephropathy (IgAN) model was duplicated by oral feeding of bovine serum albumin (BSA), subcutaneous injection of carbon tetrachloride (CCl4) and injection of lipopolysaccharide (LSP) into vena caudalis. The rats were divided into three groups randomly for the normal, IgAN model group and the group treated with Astragalus membranaceus (treatment group). The treatment group was given the Astragalus membranaceus granules via intragastric administratsion, the normal group and the IgAN model group were given the equal amount of aqua destillata by gastric perfusion. The rats were examined for albuminuria, hematuria and pathological changes of renal tissue and the distribution of TGF-beta and interleukin-5 in renal tissue was determined by immunohistochemistry and the IFN-gamma and IL-4 of cytokine of Th1 and Th2 types were detected in rats IgA nephropathy model by sandwich enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>(1) The hematuria in rats with IgA nephropathy significantly increased compared with normal control group and Astragalus treatment group (P < 0.05). There was significant increase in albuminuria in rats with IgA nephropathy, compared with normal control group and astragalus treatment group (P < 0.01). (2) The pathological change of glomerular mesangium, renal tubules and renal interstitia became serious in rats IgA nephropathy model when compared with normal control group and astragalus treatment group. Immumofluorescence showed renal IgA density in rats IgA nephropathy model was significantly higher than that in the normal control group (P < 0.001) and astragalus treatment group (P < 0.001). (3) The result of immuno histochemistry showed that there was only weak expression of TGF-beta and interleukin 5 in normal renal tissue. The expression of TGF-beta and interleukin 5 in IgA nephropathy model was significantly stronger than those in normal control group (P < 0.05) and astragalus treatment group (P < 0.05). (4) The serum IL-4 levels were (33.74 +/- 7.52) pg/ml in rats IgA nephropathy model, significantly higher than that in normal control group (2.36 +/- 0.85) pg/ml and astragalus treatment group (3.24 +/- 1.13) pg/ml. The IFN-gamma level in serum of rats IgA nephropathy model was (18.79 +/- 3.80) pg/ml, which was significantly higher than that in normal control group (46.53 +/- 5.56) pg/ml and astragalus treatment group (41.28 +/- 2.95) pg/ml.</p><p><b>CONCLUSIONS</b>The astragalus could lower the level of hematuria and 24 hours-albuminuria of the IgAN model, and amelioratse the change of the renal pathology and reduce the deposit of IgA in glomerular mesangium. The possible mechanism of the effect is that astragalus could regulate the derangement of Th1, Th2, accordingly could improve the level of IL-4 and IFN-gamma in the serum and diminish the expression of cytokine Th2 TGF-beta1 and IL-5 of the renal tissue, and thereby could postpone the development of IgAN.</p>
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Animais , Bovinos , Ratos , Astragalus propinquus , Química , Alergia e Imunologia , Medicamentos de Ervas Chinesas , Farmacologia , Glomerulonefrite por IGA , Alergia e Imunologia , Interleucina-4 , Farmacologia , Interleucina-5 , Farmacologia , Túbulos Renais , Fator de Crescimento Transformador beta , Alergia e Imunologia , Fator de Crescimento Transformador beta1 , FarmacologiaRESUMO
OBJECTIVE@#To explore the relationship between pathological features and clinical manifestations in children with nephropathy under 6 years old.@*METHODS@#Renal biopsy by rapid percutaneous puncturation was performed on 313 children under 6 who were all diagnosed clinically as kidney diseases of 14 different kinds. The specimens were divided into 3 parts for microscope, electron microscope and immuno fluorescence examination respectively and processed by HE, PAS, PASM, and Masson staining. Immunofluorescence was used to detect the deposition of IgG, IgM, IgA, C3, C4, C1q, and Fb in the renal tissues. Additional examinations were done to detect HBs-Ag, HBeAg and HBcAg deposition in some cases with positive serum HBs-Ag. Altogether 290 of the specimens (290/313, 92.65%) were examined by electron microscope.@*RESULTS@#All the renal biopsy performances were successful. The clinical manifestations comprised of persistent haematuria (32.92%, 103/313), idiopathic nephritic syndrome (26.1%, 82/313), acute nephritic syndrome (20.14%, 63/313), Henoch Schonlein purpura nephritis (8.32%, 26/313), HBV-nephritis (4.79%, 15/313), and isolated proteinuria (2.56%, 8/313). The main pathological patterns of glomerular disease were identified as mesangial proliferation (51.75%, 162/313), IgM nephropathy (8.31%,26/313), minor and minimal change (7.99%, 25/313), IgA nephropathy (7.35%, 23/313), endocapillary proliferative glomerulonephritis (5.11%, 16/313), focus segmental glomerulosclerosis (4.47%, 14/313), thin basement membrane nephropathy (4.47%, 14/313), and membrane nephropathy (4.47%, 14/313). Alport syndrome, congenital nephrotic syndrome, and thin basement membrane nephropathy can be diagnosed by electron microscope, white IgA nephropathy, IgM nephropathy and C1q nephropathy by immunopathology.@*CONCLUSION@#Similar clinical manifestations may differ in the pathology and the clinical features of one pathological diagnosis may vary greatly. Renal biopsy is of great help to the diagnosis, treatment and the prognosis evaluation for children with nephropathy under 6. Electron microscopes also play an important role in the diagnosis of nephropathy.
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Criança , Pré-Escolar , Humanos , Lactente , Biópsia por Agulha , Glomerulonefrite , Diagnóstico , Patologia , Rim , Patologia , Nefropatias , Diagnóstico , Patologia , Síndrome Nefrótica , Diagnóstico , PatologiaRESUMO
<p><b>OBJECTIVE</b>To study the mobilization effects of stem cell factor (SCF) along with granulocyte colony-stimulating factor (G-CSF) on bone marrow stem cells and endothelial progenitor cells in rats with unilateral ureteral obstruction (UUO).</p><p><b>METHODS</b>Fifty-six healthy male Wistar rats were randomly divided into seven groups: control, SCF, G-CSF, SCF+G-SCF, Sham-operated, UUO and UUO+SCF+G-CSF groups (n=8 each). The rats from the control, SCF, G-CSF and SCF+G-CSF groups were hypodermically injected with normal saline (2 mL/kg), SCF (200 microg/kg), G-CSF (200 microg/kg) and SCF along with G-CSF respectively for 5 days. The rats from the UUO and UUO+SCF+G-CSF groups were subjected to the ligation of right ureter and then were hypodermically injected with normal saline (2 mL/kg) and SCF (200 microg/kg)+G-CSF (200 microg/kg) respectively for 5 days. The sham-operated group had the same operative approach as the UUO and the UUO+SCF+G-CSF groups but the right ureter was not ligated. After operation they received a hypodermical injection of 2 mL/kg normal saline for 5 days. Five days later blood samples were collected. The percentages of CD34+ and CD34+/CD133+ cells in intravenous blood mononuclear cells were detected by flow cytometry. Serum contents of glutamate-pyruvate transaminase (GPT), glutamic oxalacetic transaminase (GOT), urea nitrogen and creatinin were measured.</p><p><b>RESULTS</b>Except for the sham-operated group, the other five groups (SCF, G-CSF, SCF+G-SCF, UUO and UUO+ SCF+G-CSF groups) had significantly higher percentage of CD34+ cells and CD34+/CD133+ cells in intravenous blood mononuclear cells than the control group (P < 0.05). There were significant differences in the percentage of CD34+ cells and CD34+/CD133+ cells among the five groups (P < 0.05). The UUO+SCF+G-CSF group showed the highest percentage of CD34+ cells and CD34+/CD133+ cells, followed by the SCF+G-CSF group. There were no significant differences in serum contents of GPT, urea nitrogen and creatinin among the seven groups. Except the UUO group showed higher GOT contents, there were no significant differences in the GOT contents among the other six groups.</p><p><b>CONCLUSIONS</b>The mobilization effects of SCF and G-CSF on bone marrow stem cells and endothelial progenitor cells were not always in paraller. A combination of SCF and G-CSF can effectively mobilize stem cells and endothelial progenitor cells, and side effects were not found in the liver and the kidney.</p>
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Animais , Masculino , Ratos , Antígeno AC133 , Antígenos CD , Antígenos CD34 , Células da Medula Óssea , Biologia Celular , Células Endoteliais , Biologia Celular , Glicoproteínas , Fator Estimulador de Colônias de Granulócitos , Farmacologia , Mobilização de Células-Tronco Hematopoéticas , Peptídeos , Ratos Wistar , Proteínas Recombinantes , Fator de Células-Tronco , Farmacologia , Obstrução UreteralRESUMO
<p><b>OBJECTIVE</b>This study investigated the clinical manifestations and renal pathological findings of 95 children with Henoch-Schonlein purpura nephritis (HSPN) in order to explore the relationship between clinical manifestations and renal pathology in HSPN.</p><p><b>METHODS</b>According to clinical manifestations, 95 HSP patients were classified into six clinical groups: 1) normal urine analysis; 2) isolated hematuria or proteinuria; 3) proteinuria with hematuria; 4) acute nephritis; 5) nephrotic syndrome; 6) acute nephritis with over 50 mg/(kg.d) of proteinuria. The severity of the renal pathological findings was determined based on the classification of the International Study of Kidney Disease (ISKDC), including grades I-VI. The relationship between clinical manifestations and the severity of renal pathological findings was studied.</p><p><b>RESULTS</b>Nephrotic syndrome was the most common clinical diagnosis (26 cases), followed by proteinuria with hematuria (23 cases), normal urine analysis (20 cases), isolated hematuria or proteinuria (15 cases), acute nephritis with over 50 mg/(kg.d) of proteinuria (7 cases) and acute nephritis (4 cases). Twenty-five out of 26 patients with nephrotic syndrome had an ISKDC classification of grade III-IV. All of the four patients with acute nephrits had a classification of grade IIIb. The 7 cases of acute nephritis with over 50 mg/(kg.d) of proteinuria had a classification of grade IIIa-V. The 20 patients with normal urine analysis had a classification of grade Iia- IIIb. There were no significant differences in ISKDC classification among the patients with normal urine analysis, isolated hematuria or proteinuria, and hematuria plus proteinuria. As the course progressed, the degree of renal pathological changes in patients with isolated hematuria or proteinuria and hematuria plus proteinuria became more serious. Of all the 95 patients, 58% had co-deposition of immunoglobulins A, G and M. The percentage of co-deposition of immunoglobulins A, G and M was related to the disease course and the severity of renal pathological findings.</p><p><b>CONCLUSIONS</b>HSPN children with nephrotic syndrome or acute nephritis with or without proteinuria had relatively severe renal pathological changes. The clinical manifestations were not always in parallel with the severity of renal pathological findings in HSPN children. With the course progressing, the renal pathological changes tended to be serious. The severe renal pathological manifestations came with co-deposition of immunogolobulins A, G and M in the glomerulin.</p>
Assuntos
Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Complexo Antígeno-Anticorpo , Rim , Patologia , Nefrite , Alergia e Imunologia , Patologia , Vasculite por IgA , Alergia e Imunologia , PatologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the role of mast cells in the development of renal interstitial fibrosis in children with Henoch-Schonlein purpura nephritis (HSPN) and possible mechanisms.</p><p><b>METHODS</b>Paraffin-embedded renal biopsy tissue sections from 20 children with HSPN were examined for the levels of tryptase-beta and transforming growth factor-beta1 (TGF-beta1) by immunohistochemical staining. Mast cells were counted by toluidine blue staining. Masson staining was used to assess the level of renal interstitial fibrosis and renal histopathological scores. Normal renal tissue sections from 5 nephrectomized children for nephroma were used as control group.</p><p><b>RESULTS</b>The percentages of positive tryptase-beta cellsand mast cells and the TGF-beta1 expression in the HSPN group were significantly higher than those in the control group (P < 0.05). The percentages of positive tryptase-beta cells and mast cells and the TGF-beta1 expression in renal tissue were positively correlated with the glomeruli histopathological score (r =0.940, 0.920, 0.937, respectively; P < 0.05) and were also positively correlated with the histopathological score of renal interstitium (r=0.903, 0.859, 0.948, respectively; P < 0.05). The level of renal interstitial fibrosis was positively correlated with the percentages of positive tryptase-beta cells and mast cells and the expression of TGF-beta1 (r =0.790, 0.766, 0.858, respectively; P < 0.05). There was a positive correlation between the percentages of positive tryptase-beta cells and mast cells (r =0.941, P < 0.05), between the percentage of positive tryptase-beta cells and the TGF-beta1 expression (r =0.897, P < 0.05) and between the percentage of positive mast cells and the TGF-beta1 expression (r=0.942, P < 0.05).</p><p><b>CONCLUSIONS</b>Tubulointerstitial mast cell infiltration is associated with the development of renal interstitial fibrosis in children with HSPN. Mast cells together with TGF-beta1 and mast cell-derived tryptase-beta may be involved in the development of the renal interstitial fibrosis in HSPN.</p>
Assuntos
Adolescente , Criança , Feminino , Humanos , Masculino , Fibrose , Rim , Química , Patologia , Mastócitos , Fisiologia , Nefrite , Patologia , Vasculite por IgA , Metabolismo , Patologia , Fator de Crescimento Transformador beta1 , TriptasesRESUMO
<p><b>OBJECTIVE</b>To investigate the clinicopathologic characteristics of childhood renal diseases.</p><p><b>METHODS</b>A retrospective analysis of 1316 renal biopsies performed over the past 20 years was performed.</p><p><b>RESULTS</b>Of the 1316 patients, 383 (29.09% ) were diagnosed as nephrotic syndrome, 291 (22.00%) as acute nephritis syndrome, 224 (17.21%) as isolated hematuria, 209(15.87%) as purpura nephritis, and 96 (7.30% ) as hepatitis B virus-associated nephritis . Mesangial proliferation was the most common pathological change (756 cases; 57.45%), followed by IgA nephropathy (113 cases; 8.59%), endothelial capillary proliferation(112 cases; 8.51%), membranous nephropathy (66 cases; 5.02%), and various minor and minimal changes (59 cases; 4.48%). Alport syndrome, congenital nephrotic syndrome, thin basement membrane nephropathy, fibrillary glomerulopathy disease, and Fabry disease were confirmed by electronic microscopy. IgA, IgM and C1q nephropathy were definitely diagnosed using immune histochemistry or immunofluorescent. A diagnosis of primary glomerular disease was made in 69.53% of the cases (915 cases); secondary glomerular disease was noted in 26.14% (344 cases). Of the 915 cases of primary glomerular disease, 375 (41.0%) had nephrotic syndrome. Secondary glomerular disease due to purura nephritis was common (209/344; 60.8%).</p><p><b>CONCLUSIONS</b>Primiary glomerular disease predominates in children. Nephrotic syndrome is the most common clinical diagnosis. Mesangial proliferation is the most common pathological patterns in children with renal disease.</p>
Assuntos
Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Rim , Patologia , Nefropatias , Patologia , Glomérulos Renais , Patologia , Insuficiência Renal , Patologia , Estudos RetrospectivosRESUMO
<p><b>OBJECTIVE</b>To investigate fibronectin synthesis in SD rat mesangial cells after transforming growth factor-beta1 (TGF-beta1) is silenced by the short interfering RNA (siRNA) expressed by reconstructed pGEFP-C1 vectors.</p><p><b>METHODS</b>Depending upon the 538th - 556th (A) and 895th - 913th (B) nucleotides of rat TGF-beta1 gene, a nucleotide (A or B) was constructed into a small hairpin nucleotide which was separately (A or B) or together (A plus B) inserted into a pGEFP-C1 vector with three reconstructed pGEFP-C1 vectors separately expressing the siRNAs for A or/and B. TGF-beta1 and fibronectin were dynamically investigated for their interrelationship by ELISA in the supernatant and RT-PCR in their extracted total RNA.</p><p><b>RESULTS</b>The siRNA hairpin-like molecules were constructed according to the 538th - 556th nucleotides of rat TGF-beta1 gene were able to markedly silence the expression of TGF-beta1 mRNA (P < 0.01) and protein (P < 0.01) at 48 h. Lipfectamin 2000 transfection stimulated the peak secretion of fibronectin at 24 h in the control and the experimental group whose TGF-beta1 was not silenced, but the silence of TGF-beta1 in both experimental groups delayed the top values of fibronectin to 48 h (P < 0.01).</p><p><b>CONCLUSION</b>The silence of TGF-beta1 by siRNA decreased the fibronectin expression, but the latter was possibly not completely TGF-dependent.</p>
Assuntos
Animais , Ratos , Células , Células Cultivadas , Fibronectinas , Metabolismo , Células Mesangiais , Metabolismo , Interferência de RNA , Fisiologia , RNA Mensageiro , Metabolismo , RNA Interferente Pequeno , Farmacologia , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1 , Química , GenéticaRESUMO
OBJECTIVE@#To evaluate the clinical and pathological features of 94 children suffering from IgA nephropathy (IgAN) while estimating the prevalent situation in Hunan province.@*METHODS@#To summarize the annual number of hospitalized children, those with kidney diseases, those accepted biopsy, and those confirmed as IgAN in both Xiangya Hospital and Second Xiangya Hospital undertaking kidney biopsy in Hunan province during 1995 and 2004.@*RESULTS@#In the past 10 years, as the hospitalized population in both hospitals accrued to 9.98% each year. The rate of 7.5% was seen in those with kidney diseases. Among whom 56.3% accepted kidney biopsy and 94 of them were confirmed as IgAN. Hematuria was the main clinical presentation, seen in 71 cases, accounting to 76%, and even to 98% after excluding those with nephrotic syndrome and isolating proteinuria type of IgAN. Inflammation infiltration (91%), renal tubule degeneration (81%), and renal interstitial fibrosis (31%) were the major pathological features of 94 children, especially in nephrotic syndrome IgAN.@*CONCLUSION@#The number of children with IgAN synchronously accrues as hospitalized population, those with kidney diseases, and those by kidney biopsy. Hematuria is the major symptom. To routinely perform urine analysis and kidney biopsy in asymptomatic hematuria may improve the diagnosis. Inflammation infiltration, renal tubule degeneration, and renal interstitial fibrosis are the major pathological features in IgAN children, especially in nephrotic syndrome IgAN, probably relating to continuous proteinuria. Early control of proteinuria may delay or decrease renal tubule fibrosis.
Assuntos
Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Biópsia por Agulha , China , Epidemiologia , Glomerulonefrite por IGA , Epidemiologia , Patologia , Hematúria , Diagnóstico , Hospitalização , Rim , PatologiaRESUMO
OBJECTIVE@#To explore the effect and possible mechanism of catechin microcapsulation on the repair of DNA damage in glumreular mesangial cells (GMCs) induced by H2O2.@*METHODS@#According to H2O2 concentration, the experiment GMCs were divided into 6 groups: a control group, 50 micromol/L group, 100 micromol/L group, 150 micromol/L group, 200 micromol/L group and 250 micromol/L group. Each group was sub-divided into 3 groups: 6 h group, 12 h group and 24 h group, in order to determining the optimum dose and the best time of detecting the DNA damage in GMCs. The cultured cells were divided into 8 groups as follows: the NS control group, the H2O2 group, the catechin groups (the final concentrations were 10.0, 15.0, and 20.0 mg/L respectively) and the various catechin microcapsulation groups (the final concentrations were 10.0, 15.0, and 20.0 mg/L respectively). At the end of the experiment, hydroxy radical (OH), malonydialdehyde (MDA) and total superoxide dismutase (tSOD) concentration of supernadant in GMCs were determined by biochemistry assay, the repair of DNA damage in GMCs were detected by single cell gel electrophoresis assay.@*RESULTS@#(1)At 6th h, H2O2 of 100 micromoL/L could cause the DNA damage of GMCs, and H2O2 of 150 micromol/L could result in DNA damage significantly. (2) No difference was found in the comet span of GMCs DNA in the catechin group and catechin microcapsulation group of different concentrations, while the DNA comet tail-long in the catechin microcapsulation group was shorter than that of the catechin group(all P(s)<0.05), and the fluorescence intensity of tail in the catechin microcapsulation group was lower than that of the catechin group(all P(s)<0.01). (3)When the concentration of catechin was 10.0 mg/L, no statistical significance was obtained in the concentration of dOH-, MDA and tSOD between the catechin microcapsulation group and the catechin group; while dOH- and MDA concentrations were lower, and the tSOD was higher in the catechin microcapsulation group than that in the catechin group when the concentration of catechin was 15.0 mg/L and 20.0 mg/L(all P(s)<0.05).@*CONCLUSION@#Catechin microcapsulation can enhance the GMCs ability of repairing DNA damage,which may be due to elevating the capacity of its anti-oxidation by catechin microcapsulation.