ABSTRACT
Objective To explore the effect of hypoxia on exosomes secreted by renal tubular epithelial cells and the function of exosomes in chronic kidney diseases.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normoxia (21％ O2) or hypoxia(1％ O2) for 48 h was collected and centrifuged gradiently to harvest exosomes.Exosomes were identified and compared by transmission electron microscope, nanoparticle tracking analysis, Western blotting and measurement of the protein concentration.(2) Primary peritoneal macrophages of rats were co-cultured with exosomes in different concentrations (1, 10, 50, 100, 300 mg/L).The expression of interleukin-6(IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) in cells and supernatant were separately detected by quantitative real-time PCR (qRT-PCR) and ELISA, and the expression of phospho (p)-STAT/STAT and suppressors of cytokine signaling 1 (SOCS1) in macrophages was detected by Western blotting.At last, the expression of inflammatory microRNAs (miR) in exosomes was measured by qRT-PCR.Results (1) The vesicles harvested by gradient centrifugation were less than 150 nm and expressed CD63 which was characteristic of exosomes.Hypoxia had no effect on the morphology of exosomes, but stimulated their secretion.(2) Hypoxic exosomes dose-dependently improved the expression of IL-6, TNF-α, iNOS in macrophages polarized by lipopolysaccharide (LPS) and increased the expression of p-STAT while decreased the expression of SOCS1 (P ＜ 0.01).MicroRNAs referred to inflammation such as miR-155 and miR-27a increased in hypoxic exosomes compared to that in normoxic exosomes (P ＜ 0.05).Conclusions Hypoxia makes exosomes promoted the polarization of macrophages to M1, which may account for the microinflammation in chronic kidney diseases.
ABSTRACT
Objective The study was aimed to detect the correlation of B cell activating factor (BAFF) promotor polymorphism-871 C/T and systemic lupus erythematosus (SLE).Methods BAFF promotor polymorphism-871 C/T was detected by the means of allele specific polymerase chain reaction (ASPCR) and agarose gel electrophoresis in 76 cases of SLE and 80 nonthrombosis normal individuals.The data of genotypic frequency and allele genotypic frequency were analyzed statistically with x2 test between the two groups.Results Fifty-one point two percent of normal individuals exhibited C/C.Thirty-five percent were heterozygous for C/T,and 13.8％ were homozygous for T/T.SLE group exhibited a different distribution pattern (30.3％ C/C,43.4％C/T,26.3％T/T).The allele frequency of T in SLE and normal individuals was 48.0％ and 31.2％ respectively.There was significant difference in the BAFF-871 C/T genotypic frequency between the SLE and nonthrombosis normal individuals (P＜0.05).Conclusion The polymorphism-871 C/T of BAFF promoter is correlated with the pathogenesis of SLE.The gene may be a major susceptible gene for SLE in Chinses Han people.Further investigations may be needed.
ABSTRACT
Famous TCM gynecological experts in the contemporary era represent the highest level of TCM gynecology during modern age.This article studies the academic thoughts of eight famous TCM gynecologic experts according to academic schools.The purpose is to seek the commonness of these academic thoughts and research their characteristics.It is extremely significant in inheriting and developing these doctors′ academic thoughts without sectarianism,which will help to guide the clinical works.