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Objective To study the gene expression profile in mice kidney with acute paraquat (PQ) poisoning and identify key genes related to renal injury.Methods A total of 20 mice (C57BL/6) were randomly (random number) divided into four groups, namely control group (group A, n =5) and poisoned groups (groups B, C, D, n =5/group).In group A, mice were administrated with distilled water (0.01 mL/g weight) while in groups B, C, D were administered with equivalent volume of PQ solution (diluted from 20％ to 0.05％ with distilled water) dissolved in distilled water via a gastric tube.Mice of group A were sacrificed immediately and mice of groups B and C at 6 h and 24 h after administration of PQ.The gene expression profile changes of kidney tissue were measured by cDNA Arraychip technology.Mice of group D were observed for mortality rate 48 h later.Results The body weights of mice decreased significantly after administration of PQ.The mortality in group D at 48 h after PQ poisoning was 100％.Compared with the control group, totally 1 792 genes with differential expression variations were identified in 6 h group and 24 h group.There were 8 key genes selected through bioinformatics analysis and they were arranged in real-time PCR: Nlrc5 , Serpinb9 , Cd40 , Rnf135 , Dhx58 , Spl 10 , Fcgrl , and Arhgef12.And then, Nlrc5 , Serpinb9 and Rnf135 were under Western blot investigation.The results of PCR and Western blot showed no significant difference to those from bioinformatics genetic analysis.Conclusions The investigation based on genome wide chip in researching related genes of PQ kidney has offered a novel idea in studying pathogenesis of acute PQ intoxication.
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Objective To investigate the effect of erythropoietin (EPO) on mesenchymal stem cells (mMSCs) proliferation under acute kidney injury (AKI) microenvironment,and to study its possible mechanism.Methods C57BL/6 mice's MSCs (mMSCs) were isolated by Percoll density gradient centrifugation and adherence cultivation.Surface markers were identified by flow cytometry.AKI mice models were made by clamping bilateral renal pedicles for 30 minutes and reopening for 30 minutes.Then both renal cortex was drew immediately to make IR kidney homogenate supernatant.P3-mMSCs were divided into different groups: Group A: low glucose DMEM medium with 10% fetal bovine serum; Group B: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant; Group C: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant and different concentrations of EPO (1,5,10,50 U/ml).Each group was incubated for 1 d,3 d,5 d,7 d.Proliferation of mMSCs was detected by CCK-8,and apoptosis was detected by TUNEL.The protein expression of erythropoietin receptor(EPOR) and the proteins of proliferation/apoptosis related signal pathway were examined by Western blotting.Results Under IR kidney homogenate supernatant,the proliferation ability of mMSCs decreased significantly (P＜0.01),while the apoptoic percentage was significantly higher than that of Group A (P＜0.01).After intervention of EPO,mMSCs proliferation enhanced,at the same time,the apoptoic percentage decreased,in a dose-dependent manner.EPOR was positive in P3-mMSCs by Western blotting.EPO decreased the expression of caspase-3 in mMSCs under AKI microenvironment in a dose- and time-dependent manner,but increased the expression of Bcl-2.Cultured for 5 d,the expression of phosphor-Janus kinase2(p-JAK2) [(0.641 ±0.028) vs (0.456±0.012)] and phosphor-signal transducer and activator of transcription(p-STAT5)[(0.398±0.016) vs (0.209±0.020)] was significantly higher in 10 U/ml EPO group compared to group B.Conclusion Erythropoietin can promote proliferation of mMSCs in vitro under AKI microenvironment,which is mediated by EPOR and related with proliferation/apoptosis signal pathway.
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Objective To investigate the application of expanded p01ytetrafluoroethylene(ePTFE)grafts in upper arm to build arteriovenous aCCeSS for hemodialysis. Methods ePTFE graft vascular access was built in the upper arm in 20 uremia patients.Three operation strategies were applied according to the reference,including loop grafts connected axillary artery and axillary vein,straight graft connected axillary artery and elbow basilic vein,and bridge connected elbow brachial artery and axillary vein. Results Twenty operations were successful and after 6-8 weeks the fistula of all cases were used in hemodialysis.The blood flows were 220-300 ml/min without re-circulation found.Conclusion ePTFE graft arteriovenous vascular access in the upper arm could be an alternative for hemodialysis patients who are difficult to build native arteriovenous fistula.
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Objective To observe the changes of CD4+CD25+ regulatory T cells, Foxp3 mRNA and soluble interlukin 2 receptor (sIL-2R) in the peripheral blood of kidney transplantation recipients and to evaluate their effect on the diagnosis of acute rejection. Methods Forty-two renal transplant recipients and 30 healthy controls were enrolled in this study. CD4+CD25+ regulatory T cells proportion, Foxp3 mRNA and sIL-2R of pre-transplantation and those of day 7,14, 28, 56 of post-transplantation were measured by flow cytometer, fluorescent quantization PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Biochemistry appliance was used to detect serum creatinine. The diagnosis of acute rejection in transplanted kidney was based on the clinical symptoms, the laboratory examinations, Doppler ultrasound and biopsy. Results (1)At day 7, 14, 28, 56 of post-transplantation, CD4+CD25+ regulatory T ceils proportion, Foxp3 mRNA level in acute rejection group were significantly decreased compared with those in non-acute rejection group. (2) There were significant differences of peripheral blood CD4+CD25+ regulatory Tcells[(9.22±3.53)% vs (6.09±1.99)%, P<0.01], Foxp3 mRNA[(0.82±0.36)×10-3 vs (0.50±0.28)×10-3, P<0.01] and sIL-2R levels [(856.30±108,24) U/ml vs (247.35±11.24) U/ml, P<0.01]between patients of pre-transplantation and healthy control group. (3)Plasma CD4+CD25+ regulatory T cells [(16.53±4.14)%] and the expression of Foxp3 mRNA [(4.97±1.94)×10-3] was significantly increased, but sIL-2R level [(463.72±31.23) U/ml] was significantly decreased as the transplanted renal function was restored (all P<0.01). (4) Plasma CD4+CD25+regulatory T cells [(12.18~2.86)%] and the expression of Foxp3 mRNA [(3.15±1.22)×10-3] was significantly decreased (P<0.01), and sIL-2R level [(748.36±115.41) U/ml] was significantly increased (P<0.01) when acute rejection occurred. The above changes had an earlier onset than the change of Scr. (5)The percentage of CD4+CD25+ regulatory T cells was positively correlated with the Foxp3 mRNA level (P<0.01), but was not correlated with sIL-2R level in all the patients. Conclusion The measurement of these markers in peripheral blood may be an important guideline to the diagnosis and prognosis of acute rejection in renal transplant recipients.