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.

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.

Objective:To study the anti-fibrogenetic effect of matrine compound injection and its mechanism. Methods: Experimental liver fibrosis induced by tetrachloride in rats was treated by matrine compound injection. Liver function index, interleukin-1? (IL-1?) and pathological changes in liver tissue were observed in model and therapy groups during the 4th,6th and 9th week. Results:The serum IL-1? levels of model and therapy groups were (25. 51?14. 31) and (10. 49?7. 49) pg/ml in the 4th week, (109. 67?20. 87) and (15. 06?2. 65) pg/ml in the 6th week, and (40. 26?10. 63) and (23. 27?7. 56) pg/ml in the 9th week. The liver function (ALT, AST, TBil) of therapy group was better than that of model group at each stage. Pathological changes indicated that the infiltration of inflammatory cells,the degree of liver cell necrosis and the degree of fi-broplastic proliferation in the therapy group were obviously better than those of the model group. Conclusion : Matrine compound injection has an anti-fibrogenetic action on the liver fibrosis induced by tetrachloride.

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.

Objective To investigate the mechanism and effect of the spinal cord outlet of the skull base on Chiari Ⅰ malformation with syringomyelia.Methods The cervical spinal cord stem angle (Anbc),slope angle of cervical vertebra (Ansc) of Chiari Ⅰ malformation were measured.In foramen magnum (Llf) and anterior vertebral canal level (Laf),spinal canal(Ac),spinal cord (As) and inferior hernia area (Ah) were measured.Angle,area and ratio were compared in Chiari Ⅰ malformation with syringomyelia,Chiari Ⅰ malformation without syringomyelia and normal control group.Results Ansc,Anbc-Ansc had significant differences among control group and Chiari Ⅰ malformation patients (all P＜0.001).In Llf,Laf,As had significant differences among three groups (all P＜0.05),further comparison of the two showed there were significant differences between Chiari Ⅰ malformation with syringomyelia patients and control group,Chiari Ⅰ malformation without syringomyelia patients and control group in Llf(all P＜0.05).In Llf,Laf,Ac in Chiari Ⅰ malformation with syringomyelia was smaller than control group (P＜0.05).Ah in Llf,Lafand Lh in Llf had no statistical significant difference between Chiari Ⅰ malformation with and without syringomyelia patients (all P＞0.05).In Llf,Laf,As/Ac had statistical significant difference among Chiari Ⅰ malformation with and without syringomyelia patients,control group (all P＜0.001),further comparison of the two showed As/Ac in Llf had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group (P＜0.05),As/Ac in La had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group,between Chiari Ⅰ malformation without syringomyelia patients and control group (all P＜0.05),Conclusion The cervical spinal cord,Ansc reducing,narrow vertebral proportion increase are important factors to promote Chiari Ⅰ malformation syringomyelia.

Objective To study the effect and mechanism of bone marrow mesenchymal stem cells (BMSCs) on autologous peripheral blood lymphocyte proliferation in vitro in patients with hepatitis B virus related decompensated cirrhosis.Methods MSCs were isolated and expanded from human bone marrow blood of nine patients with decompensated cirrhosis,four groups were designed for experiment:①BMSCs + lymphocytes + PHA (contact co-culture) ; ② BMSCs + lymphocytes + PHA (non-contact co-culture) ; ③lymphocytes + PHA (positive control) ; ④lymphocytes alone (negative control).Lymphocytes proliferation rate were detected by flow cytometry.The mRNA expression levels of interleukin-10 (IL-10)and transforming growth factor-β1 (TGF-β1)in BMSCs were detected by using reverse transcription-polymerase chain reaction (RT-PCR).Results Cell proliferation of contact and non-contact co-culture groups significantly declined when compared with that of positive control group (all P ＜ 0.01),The relative expression levels of IL-10 mRNA and TGF-βlmRNA in BMSCs of contact and non-contact co-culture group raised up significantly after culture (all P ＜ 0.01).Besides,there was no significant difference on lymphocyte proliferation rate or expression levels of IL-10mRNA and TGF-β1 mRNA between contact co-culture group and non-contact coculture group (all P ＞ 0.05).Conclusion BMSCs from cirrhotic patients can inhibit proliferation of autologous peripheral blood lymphocytes,the mechanism may be associated with the secretion of inhibitory cytokine IL-10 and TGF-β1 in BMSCs.

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.

Objective To study the effect of bone marrow mesenchymal stem cells (BMSCs) on autologous peripheral blood lymphocyte proliferation and regulatory T cells (Tregs)subsets in vitro in patients with hepatitis B virus related decompensated cirrhosis.Methods MSCs were isolated and expanded from human bone marrow blood of eight patients with decompensated cirrhosis.The purity of MSCs was identified by flow cytometry.lymphocytes were isolated from the peripheral blood of patients and stained with carboxy fluoresce indiacetate succinimidyl ester (CFDA SE).The BMSCs and peripheral blood lymphocytes from patients were added into wells containing autologous serum in the presence of phytohemagglutinin (PHA).lymphocytes proliferation rate and CD4 + CD25 + CD127-Tregs frequency were detected by flow cytometry.Results Flowcytometric analysis showed that lymphocyte proliferation in contact co-culture group and noncontact co-culture group were much lower than positive control (all P ＜ 0.01),and significantly higher than negative control (all P ＜ 0.01),while CD4+ CD25+ CD127-Tregs in contact co-culture group and noncontact co-culture group were much higher than positive control and negative control (all P ＜ 0.01).Besides,there was no significant difference on lymphocyte proliferation rate or Tregs frequency between contact co-culture group and non-contact co-culture group(all P ＞ 0.05).Conclusion BMSCs can inhibit proliferation of autologous peripheral blood lymphocytes and increase expression of CD4 + CD25 + CD127-Tregsin patients with decompensated cirrhosis.

OBJECTIVE To establish a clinical pharmaceutical pathway of anti-infective therapy for high-risk populations of antibiotic-associated encephalopathy （AAE）， and analyze its application effects. METHODS Clinical pharmacists developed the “AAE High-Risk Population Screening Form” and “Antibiotic AAE Risk Comparison Form” based on literature and expert opinions， and established the “Clinical Pharmaceutical Pathway of Anti-infective Treatment for AAE High-Risk Population” in our hospital. A prospective， non-randomized controlled study was conducted from May 2023 to April 2024， including 50 cases in the observation group and 50 cases in the control group among patients with pulmonary infections admitted to the Dept. of Internal Medicine in our hospital. The observation group was involved in the development of an anti-infective treatment following the clinical pharmaceutical pathway by clinical pharmacists， while the control group received routine anti-infective treatment by clinical physicians. The occurrence of AAE， the rational antibiotic drug use， and the effectiveness of initial anti-infective treatment in the two groups were observed， and the intervention measures and outcomes of AAE cases were summarized. RESULTS The anti-infective treatment clinical pharmaceutical pathway for AAE high-risk population was preliminarily established in our hospital. The analysis of the application effects showed that there was 1 case of AAE in the observation group and 8 cases in the control group， with a significantly lower incidence of AAE in the observation group than in the control group； the rational antibiotic drug use and the effectiveness of initial anti-infective treatment in the observation group were both significantly superior to those in the control group （P＜0.05）. Drug withdrawal and dressing change were the preferred effective intervention measures for AAE， and encephalopathy treatment drugs could be used as auxiliary measures for symptom relief. Timely and effective intervention was conducive to rapid symptom relief， with a total improvement rate of AAE of 88.89%. CONCLUSIONS The anti-infective treatment clinical pharmaceutical pathway for AAE high-risk population can effectively prevent the occurrence of AAE as well as contribute to promoting rational drug use and the effectiveness of initial anti-infection plans and strengthening treatment outcomes.