AIM: To investigate the expression profile of peripheral-type benzodiazepine receptor(PBR) involved in mitochondrial permeability transition(PT) regulation,and to observe the binding dynamic of the mitochondrial PBR with specificity ligand during rat live regeneration.METHODS: Liver regeneration model was produced by 70% partial hepatectomy(PH) performed in male SD rats.The animals of sham groups underwent the same surgical operations as PH groups did,but the liver lobes were not resected.The animals in the PH groups and corresponding sham groups were sacrificed at 3,6,12,24,48,72,120 and 168 hours after the operation.The livers were removed,weighted and processed for isolation of mitochondria.Semi-quantitative RT-PCR was performed to examine the expression level of PBR in 70% hepatectomized rat livers during the whole regeneration process and compared to that in the sham and normal groups.Compared with healthy rats,the kinetic parameters of PBR was evaluated by using a specific radioligand -PK11195.RESULTS: Compared with healthy rats,the expression of PBR was unchanged.Meanwhile,the results obtained in the present experiments by scatchard analysis,Bmax of PK11195 for PBR significantly decreased,returned to normal level in 168 h after PH.Kd of PK11195 for PBR significantly decreased at 72 h and 168 h after PH of rat liver regeneration(P

Objective:To analyze the association between CRHR1 gene(rs1876828)polymorphism and the effect of inhaled corticosteroids(ICS)in children with bronchial asthma.Methods:A total of 60 children with moderate persistent asthma who were treated in the Department of Pediatrics of the First Affiliated Hospital of Harbin Medical University from January 2018 to June 2019 were included.The CRHR1 gene rs1876828 locus in children with asthma was detected by Sanger sequencing.The children were divided into TT genotype group(TT group) and CC genotype group(CC group)according to the different base sequences of gene loci.There were 22 cases in TT group(36.7%)and 38 cases in CC group(63.3%). Both groups were given aerosol inhalation of ICS and symptomatic treatment.Clinical symptoms and signs were observed and scored before and after treatment for 3d, 10d and 30d, and the days required for complete disappearance of symptoms and signs were recorded.Results:After 3d of treatment, clinical symptoms and signs of TT group and CC group were improved to a certain extent, but there was no statistical significant difference between two groups( P>0.05). At 10d and 30 d after treatment, the recovery of the two groups was better than that at 3d, and the improvement degree of the TT group was significantly better than that of the CC group, with statistical significance( P<0.05). The time of complete remission of symptoms and signs in TT group and CC group was(8.68±7.42)d and(16.21±7.82)d; the difference was statistically significant( P<0.01). Conclusion:There is a polymorphism of CRHR1 rs1876828 locus in children with bronchial asthma, which manifests as TT genotype and CC genotype, and CC genotype is the majority.The polymorphism of CRHR1 gene rs1876828 in asthmatic children is associated with the efficacy of ICS.The efficacy of ICS in children with TT genotype is better than that of CC genotype.

Objective:To investigate the regulatory effects and mechanism of mannose-binding lectin(MBL) on autophagy during the differentiation of 3T3-L1 adipocytes, and provide the feasibility for targeting autophagy to prevent obesity and related pathological conditions in natural immunity.Methods:3T3-L1 preadipocytes were cultured in vitro and induced to differentiation. Cell differentiation and lipid accumulation were analyzed by oil red O staining and CCK-8 was used to detect the effect of different concentrations of MBL (0, 1, 5, 10 μg/ml) on cell proliferation ability at different differentiation stages. Western blot was used to analyze the expression of MBL(10 μg/ml) on the key autophagy factors LC3B, Beclin1 and p62 protein at different stages of differentiation, and the changes of lipid droplet accumulation under the intervention of MBL were observed by oil red O staining. The protein and mRNA expression of autophagy key factors under the intervention of different concentrations of MBL were detected by Western blot and qRT-PCR. And autophagy flow analysis based on autophagic degradation was used to further illustrate the autophagic activity. The expression and phosphorylation of adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) signaling molecules were analyzed by Western blot. Results:The results of oil red O staining showed that 3T3-L1 preadipocytes could achieve complete differentiation after 10 days of induction. CCK-8 showed that the concentration of MBL (1-10 μg/ml) in the experimental group had no effect on cell proliferation at different differentiation stages. During the differentiation of 3T3-L1 preadipocytes, Western blot and qRT-PCR showed that the expression of autophagy-related proteins and mRNA levels was enhanced in the MBL treated group, and presented a concentration-dependent relationship. Oil red O staining showed that the lipid droplets in adipocytes at different stages of differentiation are reduced to varying degrees under the intervention of MBL. Fluorescence microscopy results further confirmed that MBL enhanced the autophagy activity of adipocytes by increasing the synthesis of autophagosomes. Moreover, under the intervention of MBL, the phosphorylation level of AMPK was significantly up-regulated, while the phosphorylation level of mTOR was significantly down-regulated, also showing a concentration-dependent relationship.Conclusions:MBL accelerates the autophagy process during the differentiation of 3T3-L1 adipocytes through AMPK/mTOR signaling pathway, reduces lipid accumulation, providing a possible functional pathway for the treatment of obesity and related metabolic diseases.