ABSTRACT
AIM:To observe the expression of Snail1 and insulin-like growth factor-1 (IGF-1) in NRK-52E cells induced by high glucose,and to investigate the relationship of Snail1 and IGF-1 in the mechanism of epithelial to mesenchymal transition (EMT) in diabetic kidney disease (DKD).METHODS:The NRK-52E cells were treated with Snail1 siRNA and IGF-1 siRNA after cultured with high glucose medium for 72 h,and divided into control group,high glucose group,non-targeting (NT) siRNA group,Snail1 RNAi group and IGF-1 RNAi group.The cells were harvested at 48 h and 72 h.Real-time PCR was used to detect the mRNA expression of Snail1,IGF-1,E-cadherin and fibronectin (FN),and the protein levels were determined by immunofluorescence staining.RESULTS:Compared with control group,the expression of E-cadherin at mRNA and protein levels declined after stimulation with high glucose (P ＜ 0.01),while that of FN was elevated (P ＜0.01).Meanwhile,the mRNA and protein levels of Snail1 and IGF-1 were markedly increased (P ＜0.01).The expression of E-cadherin at mRNA and protein levels was improved in Snail1 RNAi group as compared with high glucose group (P ＜ 0.01),while that of FN,IGF-l and Snail1 was significantly down-regulated (P ＜ 0.01).The same changes were observed in IGF-1 RNAi group (P ＜0.01).The protein expression of each factor in NT group had no significant change as compared with high glucose group (P ＞ 0.05).Pearson correlation analysis showed a close positive relationship between the expression of Snail1 and IGF-1 protein (r =0.852,P ＜ 0.01).CONCLUSION:Snail1 may facilitate DKD development by regulating IGF-1 in the process of EMT.
ABSTRACT
AIM:To observe the expression of Snail1 and insulin-like growth factor-1 (IGF-1) in NRK-52E cells induced by high glucose,and to investigate the relationship of Snail1 and IGF-1 in the mechanism of epithelial to mesenchymal transition (EMT) in diabetic kidney disease (DKD).METHODS:The NRK-52E cells were treated with Snail1 siRNA and IGF-1 siRNA after cultured with high glucose medium for 72 h,and divided into control group,high glucose group,non-targeting (NT) siRNA group,Snail1 RNAi group and IGF-1 RNAi group.The cells were harvested at 48 h and 72 h.Real-time PCR was used to detect the mRNA expression of Snail1,IGF-1,E-cadherin and fibronectin (FN),and the protein levels were determined by immunofluorescence staining.RESULTS:Compared with control group,the expression of E-cadherin at mRNA and protein levels declined after stimulation with high glucose (P ＜ 0.01),while that of FN was elevated (P ＜0.01).Meanwhile,the mRNA and protein levels of Snail1 and IGF-1 were markedly increased (P ＜0.01).The expression of E-cadherin at mRNA and protein levels was improved in Snail1 RNAi group as compared with high glucose group (P ＜ 0.01),while that of FN,IGF-l and Snail1 was significantly down-regulated (P ＜ 0.01).The same changes were observed in IGF-1 RNAi group (P ＜0.01).The protein expression of each factor in NT group had no significant change as compared with high glucose group (P ＞ 0.05).Pearson correlation analysis showed a close positive relationship between the expression of Snail1 and IGF-1 protein (r =0.852,P ＜ 0.01).CONCLUSION:Snail1 may facilitate DKD development by regulating IGF-1 in the process of EMT.
ABSTRACT
This study was aimed to investigate a quality control method for ABO typing of neonatal umbilical cord blood(UCB). The routine serology method was used to identify the ABO type of UCB samples. These samples with questions were further detected by sequence specific primer PCR (PCR-SSP). The results showed that among total of 76120 UCB samples identified by positive ABO typing, there were 78 samples (1 per thousand) which could not be determined. Of these 78 samples, 30 (56.92%) samples with a weak agglutination reaction were excluded by reverse ABO typing. Out of 260 samples in reverse ABO typing, 148 samples were consistent with positive ABO typing, 112 samples (43.08%) were inconsistent with the positive ABO typing. 58 undetermined samples were detected by PCR-SSP. Out of them the genotyping results of 45 samples confirmed the serological typing, the phenotyping results in 3 cases were inconsistent to that of genotyping. 10 cases showed the unconformity between positive and reverse typing, but the genotyping results were fully consistent with the positive typing. In conclusion, positive typing for red cell antigens combined with PCR-SSP is efficient and sensitive for quality control of ABO typing for neonatal UCB.
Subject(s)
Humans , Infant, Newborn , ABO Blood-Group System , Genetics , Blood Grouping and Crossmatching , Methods , Fetal Blood , Genotype , Quality ControlABSTRACT
To investigate the correlation between the HLA genes and pathogenesis of aplastic anemia (AA), polymerase chain reaction with specific sequence primers (PCR-SSP) method was used to HLA typing in 82 patients with AA and 400 normal healthy individuals as control. The results showed that A*2301 (1.84%), B*5501 (4.36%) and DRB1*0901 (23.48%) gene frequency in AA patients were significantly higher than those in controls (relative risk: RR=5.0253, 3.3645, 2.1269, chi2=4.6634, 6.3120, 9.1511 respectively) (p<0.01). In contrast, DRB1*1301 (1.23%) gene frequency was significantly lower in AA than that in controls, RR=0.2257, chi2=6.6629 (p<0.01). It is concluded that A*2301, B*5501 and DRB1*0901 genes may be considered as the risk markers while DRB1*1301 gene as a protective marker of AA.