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Objective To investigate the protective effect of protein kinase C (PKC) inhibitor rottlerin on rat renal vascular endothelial injury induced by lipopolysaccharide (LPS). Methods Rat renal microvascular endothelial cells cultured for 3-6 generations were divided into three groups according to random number table: blank control group in which cells were not challenged, LPS group in which cells were only stimulated by LPS 10 mg/L for 24 hours, and PKC inhibitor group in which cells were treated with PKC inhibitor rottlerin 2 μmol/L 30 minutes before LPS stimulation. The levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-1β, IL-8) were determined by enzyme-linked immunosorbent assay (ELISA). Monolayer permeability was determined by Transwell assay. The expressions of PKC, RhoA and vascular endothelial-cadherin (VE-cadherin) were detected by Western Blot. The morphological characteristic and distribution of F-actin was measured by laser confocal fluorescence microscope. Results Compared with blank control group, the levels of inflammatory cytokines at 24 hours after 10 mg/L LPS stimulation were significantly increased in LPS group [TNF-α (ng/L): 397.3±25.4 vs. 46.8±8.9, IL-1β(ng/L): 76.7±11.2 vs. 12.6±3.2, IL-8 (ng/L): 574.5±31.4 vs. 73.2±9.6, all P < 0.05], the permeability of endothelial cells was significantly increased (A value: 1.32±0.03 vs. 0.36±0.02, P < 0.05), while the expressions of PKC and RhoA were significantly up-regulated (PKC/β-actin: 0.88±0.02 vs. 0.61±0.03, RhoA/β-actin: 0.96±0.01 vs. 0.49±0.03, both P < 0.05), VE-cadherin expression was significantly down-regulated (VE-cadherin/β-actin: 0.51±0.01 vs. 0.72±0.04, P < 0.05), and the F-actin distribution disorder had obvious stress fiber formation. Compared with LPS group, the levels of inflammatory cytokines were significantly lowered in PKC inhibitor group [TNF-α (ng/L): 127.4±14.6 vs. 397.3±25.4, IL-1β(ng/L): 43.2±7.8 vs. 76.7±11.2, IL-8 (ng/L): 212.7±18.2 vs. 574.5±31.4, all P < 0.05], the permeability of endothelial cells was significantly decreased (A value: 0.81±0.02 vs. 1.32±0.03, P < 0.05), the expressions of PKC and RhoA were significantly down-regulated (PKC/β-actin: 0.44±0.03 vs. 0.88±0.02, RhoA/β-actin: 0.63±0.05 vs. 0.96±0.01, both P < 0.05), the VE-cadherin expression was significantly up-regulated (VE-cadherin/β-actin: 0.69±0.03 vs. 0.51±0.01, P < 0.05), and the F-actin remodeling and stress fiber formation were significantly reduced. Conclusion PKC inhibitor could significantly attenuate the damage of vascular endothelial barrier induced by LPS, and plays an important role in endothelial cell barrier.
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Objective To explore the clinical and genetic features of Rubinstein-Taybi syndrome (RSTS). Methods The clinical data of 2 children with RSTS were reviewed and analyzed. Results Two male children (3 years old and 4 months old) were admitted to hospital because of growth retardation. Both of them were characterized by short stature, language and motor retardation, excessive hairiness and cryptorchidism. Case 1 had slightly broad thumbs and toes, and case 2 had distinctive facial features of high arched palate, broad nasal bridge, ptosis, and obviously broad thumbs and toes. Cardiac dysplasia was found in both of them by echocardiography. The c.152T>G (L51X) heterozygous mutation was found in case 1 by high throughput sequencing and genomic chip technology, and this mutation has not been reported. Deletion of 2.5 Mb in chromosome 16p13.3 region was found in case 2. Conclusions The main clinical manifestations of RSTS are excess hair, deformity of thumbs and toes, deformity of the heart development, and growth retardation. Molecular detection can help the clinical diagnosis.
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Objective To confirm the role of human parvovirus B 19 ( B19) infection in childhood immune thrombocytopenia ( ITP) patients.Methods A total of 416 cases of newly diagnosed childhood ITP patients from Jan .2011 to Dec.2013 had been summarized to be cases group , Then a total of 130 childhood patients with common respiratory tract infection were selected randomly as the control group . All patients had been divided in grouped by age as 0.05) was found.All the ITP patients had not been given anti -B19 treatment.The PLT remission rate,respectively, after treated with the same pro-tocol including glucocorticoid and/or immunoglobulin had a declining trend as the ages increasing .The B19 infection groups of all ages al-so had no significant difference PLT remission rate had been confirmed in non -B19 infection patients in each age groups (all P>0.05). Conclusions B19 infection may not be a major cause in childhood newly ITP , and treatment protocol with no anti -B19 treatment had no influence on the clinical curative efficacy .
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ObjectiveTo investigate the association between nucleotide polymorphisms (SNP) of rs2282430 and rs2031234 inSLC26A9 gene and clinical characteristics of asthma in Han children in central China.MethodsA case-control study was performed. Two hundreds and three children with asthma were recruited in this study and 221 normal children were selected as controls. The genotypes of two SNPs inSLC26A9 gene were examined using PCR-RFLP.ResultsBetween children with asthma and controls, the distribution of three genotypes (AA, AG and GG) in rs2282430 locus had signiifcant difference (P=0.042). The percentage of AA genotype was higher in children with asthma than that in controls. In implicit mode (AAvs. AG+GG), the two groups was statistically signiifcant difference (P=0.028). The frequency of A allele was higher in children with asthma than that in controls (P=0.011). Between children with asthma and controls, the distribution of three genotypes (TT, GT, and GG) in rs12031234 locus had no signiifcant difference (P=0.479). The frequency of alleles in rs12031234 locus also had no signiifcant difference (P=0.215). Among asthmatic children with different genotype of rs2282430, the lymphocytecounts (LYM), C-reaction protein (CRP), IgE, neutrophils (NEU%), and eosinophils (EOS%) were not signiifcantly different (P>0.05). Conclu-sionsThe rs2282430 polymorphism inSLC26A9 gene is associated with childhood asthma in the central China and the A allele is the risk factor. The rs2282430 polymorphism is not associated with LYM counts, CRP level, IgE level, NEU%, and EOS%.