ABSTRACT
Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
Subject(s)
Humans , China , Cystic Fibrosis/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Mutation , Poly T , RNA, Messenger/geneticsABSTRACT
AIM: To observe the effects of H_2O_2 on apoptosis of skeletal muscle satellite cells (SMSC)and mitochondrial membrane potential (MMP), and protective effect of erythropoietin(EPO). METHODS: SMSC in vitro were divided into three groups: H_2O_2 group, H_2O_2+EPO group and control. Apoptosis rate and the means were obverted by monofluorescence flow cytometry. The morphological change of apoptosis cells were observed under fluorescence microscopy after Hoechst 33258 staining. RESULTS: The cells in H_2O_2 group show the highest apoptosis rate (22.13±1.79)%. In H_2O_2+EPO group, apoptosis rate were (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% according to the EPO treated levels (10, 20 or 40 kU/L), respectively. MMP level in H_2O_2 group was the lowest 9.70±0.09. MMP levels in H_2O_2+EPO group were 12.67±0.32, 27.90±0.66, 44.53±0.93, respectively according to the EPO treated levels (10, 20 or 40 kU/L). In control group, apoptosis rate was 1.93±0.57 and MMP was 51.37±0.64. In H_2O_2 group and H_2O_2+ low dosage EPO group, Hoechst 33258 staining showed obvious apoptosis. CONCLUSION: EPO inhibits the apoptosis induced by H_2O_2 and stabilizes the MMP, which is related to the dosage of EPO.
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AIM and METHODS: Chemotactic migration of peripheral blood mononuclear cell(PBMNC) of healthy blood donors(BD) and patients with blood stasis syndrome(BSS) across polycarbonate membrane(PCM) and human umbilical vein endothelial cell(HUVEC) monolayer, IL-8 produced by migrat PBMNC and effects of protocatechuic aldehyde(PCA) on the process mentioned above were investigated. RESULTS: 1) The numbers of migrating PBMNC in group BSS was higher than that in group BD(P
ABSTRACT
Objective The self-microemulsifying drug delivery system(SMEDDS) with tanshinone ⅡA was prepared in order to develop its new dosage forms.Methods Pseudo ternary phase diagrams were used to evaluate the self-microemulsification existence area under emulsifier,coemulsifer,and oil phase.The HPLC analysis in vitro was set up.Solubility in various vehicles was determined.The self-microemulsification efficiency was assessed,such as stability,particle size,and Zeta potential.Results The solubi-lity of tanshinone ⅡA in SMEDDS was about 2.5 mg/g,droplet size was within 20 nm,and the absolute value of Zeta potential was over 60 mV.The stability of SMEDDS with tanshinone ⅡA was better in centrifugal condition with high temperature but not good enough to the light.Conclusion The SMEDDS can make tanshinone ⅡA solublized in water,and is an optimum vehicle in new dosage forms of tanshinone ⅡA.
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Objective To study the puncture method of Angioseal closure device by femoral artery. Methods A prospective trial was carried out in 80 patients using Angioseal closure device in angiography and angioplasty. All patients were divided into tow groups, according to the horizontal distance between the entrance of skin and the entrance of the femoral artery: Group A, the distance 1.5 cm. Results 1. The rate of successful puncture was 92% in group A and 81% in group B (P