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Objectives To explore the association between the single nucleotide polymorphism (SNP) rs2239669 in makorin ring-finger protein 3 (MKRN3) gene and the susceptibility to central precocious puberty (CPP). Methods A case-control study including 246 children with CPP and 269 healthy children was performed.The genotype and MKRN3 expression levels of patients were analyzed by PCR-HRM and RT-PCR,respectively. Results SNP rs2239669 genotype (TT,TC,CC) and allele frequencies (T and C) were different between cases and controls,with higher CC genotype in CPP patients. Under recessive model (CC/TT+TC),CC genotype was higher in CPP group and associated with higher risk of CPP (95%CI:1.062-2.143,P=0.021). MKRN3 expression levels were different among patients with different genotypes,of which TT genotype had the highest level followed by TC and CC (0.376±0.094, 0.330±0.068, 0.250±0.072, P=0.041). Conclusions MKRN3 SNP rs2239669 was associated with increased risk of CPP, and patients with TT genotype had higher MKRN3 levels.
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Objective To investigate the molecular mechanism of calreticulin ( CRT) transcription induced by HBV and its viral proteins. Methods The human hepatocellular cell line, HepG2, was trans-fected with pHBV1. 3 and eukaryotic expression plasmids of HBV viral proteins, respectively. The expres-sion of CRT was measured after transfection. A reporter plasmid of CRT promoter was constructed to analyze the induction of CRT promoter by pHBV1. 3 and HBV viral proteins. Furthermore, two truncated and one C/EBPα site deficient mutants were constructed to evaluate the regulatory effects of HBx on CRT promoter. Fi-nally, HepG2 cells were transfected with HBx expression plasmids and the cellular localization of C/EBPαwas analyzed. Results In this study, pHBV1. 3 could significantly up-regulate the expression of CRT at mRNA and protein levels as well as enhancing the activity of CRT promoter. Among the seven HBV viral proteins, HBx could enhance the activity of CRT promoter and the expression of CRT at mRNA and protein levels. HBx could not induce the transcription of CRT when the C/EBPα binding site was deleted from the CRT promoter. The expression of HBx could promote the nuclear translocation of C/EBPα. Conclusion HBV and its viral protein HBx could up-regulate the CRT expression at transcriptional level. The transcrip-tional factor C/EBPα played a critical role in HBx-induced transcriptional activation of CRT.
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Objectives To explore the clinical features and diagnosis of LMNA-associated congenital muscular dystrophy. Methods The clinical data from a case of muscular dystrophy caused by LMNA gene mutation were retrospectively analyzed. The related literatures were reviewed. Results A 8-month-old female infant suffered from weakness of raising head, eyelid droop, and motor development retardtion. LMNA gene was sequenced for the infant, her parents and the older sister. Heterozygous mutation of c. 94_96 del AAG (p. K 32 del) was found in the infant leading to the diagnosis of LMNA- associated congenital muscular dystrophy. No mutation was found in the infant’s parents and her older sister. The literature review showed that all ofLMNA- associated congenital muscular dystrophy patients had LMNA gene mutation, more than 80% patients mainly presented with weakness of raising head and may accompany with weakness of proximal limb, motor development retardation, and weakness of axial muscle. Conclusions Mutation analysis of LMNA gene is conducive to the diagnosis of congenital muscular dystrophy.
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Objective:To investigate the role of vitamin D-induced autophagy in macrophages against Mycobacterium tuberculosis(Mtb).Methods:Induce U937 cells to differentiate into macrophages with phagocytic capacity by phorbol 12-myristate 13-acetate(PMA).The cells were randomly divided into negative control group,vitamin D group,autophagy inhibitor(3-MA)+vitamin D group,positive control(rapamycin)group.Infect all groups with Mtb for 6 hours.In the 4th day after infection,the mRNA expressions of autophagy-related genes ATG5, Beclin-1 and LL-37, LC3B were determined by semi-quantitative RT-PCR.LC3B-Ⅱ+and/or Mycobacterium tuberculosis antigen 85A+( Ag85A+)-cells were counted by flow cytometry.Results:Compared with the control groups, the mRNA expressions of ATG5,Beclin-1,LL-37 and LC3B were enhanced(P<0.01),the numbers of LC3B-Ⅱ+-cells increased,the numbers of Ag85A+-cells decreased, and the numbers of LC3B-Ⅱ+-Ag85A--cells increased ( vitamin D group 38.0% vs negative control group1.08%).Compared with the vitamin D group,the mRNA expressions of ATG5,Beclin-1 and LC3B were suppressed in the autophagy inhibitor(3-MA)+Vitamin D group,the mRNA expressions of LL-37 were reduced,and 3-MA inhibited the expression of LC3B-Ⅱ in cells with inhibition of LC3B-Ⅱ+-Ag85A--cells increase as well.Conclusion: Vitamin D can induce macrophage autophagy and further contribute to scavenging Mycobacterium tuberculosis.