Objective:To explore the effects of quercetin on autophagy and proliferation in HBV-positive liver cancer HCCLM3 cells based on STING signaling and its underlying mechanism.Methods:HCCLM3 cells were treated with quercetin (50 μmol/L or 100 μmol/L), designated as the 50 μmol/L group and 100 μmol/L group, respectively. The inhibitory effect of quercetin on HCCLM3 cells was detected using the CCK-8 method. A scratch assay was conducted to assess the impact of quercetin on the migration ability of HCCLM3 cells. A CCK8 and ROS kit was used to detect the effect of quercetin on the levels of reactive oxygen species in HCCLM3 cells. Western blotting was employed to measure the effect of quercetin on the expression of STING signaling and autophagy-related proteins in HCCLM3 cells. RNA interference technology was used to assess the effects of STING signaling inhibition on the expression of autophagy-related proteins and reactive oxygen species levels in HCCLM3 cells. The combined effects of STING activators and quercetin on HCCLM3 cell proliferation and autophagy were evaluated. The t-test was used to detect data differences between two groups, while ANOVA was employed for comparisons among multiple groups, followed by the SNK- q test for further pairwise comparisons. Results:Compared with the control group, quercetin (50 μmol/L and 100 μmol/L groups) significantly inhibited HCCLM3 cell survival activity in a dose-dependent manner (control group: 100%; 50 μmol/L group: 75.25%; 100 μmol/L group: 50.36%, P<0.01 ). Quercetin inhibited HCCLM3 cell migration in a dose-dependent manner (>2 h, control group: 187.16 μm; 50 μmol/L group: 145.22 μm; 100 μmol/L group: 88.21 μm, P<0.01), which significantly increased intracellular reactive oxygen species (ROS) levels in HCCLM3 cells (control group: 1.00; 50 μmol/L group: 1.565; 100 μmol/L group: 2.175, P<0.01). The phosphorylation level of STING was significantly increased ( P<0.01), and the expression of autophagy-related protein microtubule-related protein 1A/1B light chain 3 (LC3) protein was significantly promoted ( P<0.01). Compared with the quercetin group, the cell viability of the small interfering-STING+quercetin group was increased (quercetin group: 56.3%; small interfering-STING+quercetin group: 85.7%, P<0.05), while the expression of autophagy-related protein LC3 was decreased. Compared with the quercetin group, the cell viability of the quercetin+STING activator group was further decreased (quercetin group: 56.7%; quercetin+STING activator group: 35.4%, P<0.01), and the expression levels of STING and autophagy protein LC3 were significantly increased ( P<0.05). Conclusions:STING signaling-regulated cell autophagy mediates the inhibitory effect of quercetin on the proliferation of HCCLM3 cells, and this effect is enhanced after administration of the STING agonist.

Ribosome is an intracellular ribonucleoprotein particle that serves as the site of protein biosynthesis. Ribosomal dysfunction caused by mutations in genes encoding ribosomal proteins (RPs) and ribosome biogenesis factors (RBFs) can lead to a spectrum of diseases, collectively known as ribosomopathy. Phase separation is a thermodynamic process that produces multiple phases from a homogeneous mixture. The formation of membraneless organelles and intracellular structures, including ribosomes and nucleoli, cannot occur without the involvement of phase separation. Here, ribosome structure, biogenesis, and their relationship with ribosomopathy are systematically reviewed. The tissue specificity of ribosomopathy and the role of phase separation in ribosomopathy are particularly discussed, which may offer some clues for understanding the mechanisms of ribosomopathy. Then, some new ideas for the prevention, diagnosis, and treatment of ribosomopathy are provided.
Humans ; Ribosomes/physiology* ; Ribosomal Proteins/metabolism* ; Mutation ; Animals ; Cell Nucleolus/metabolism* ; Protein Biosynthesis ; Phase Separation

Objective:To explore the effects of quercetin on autophagy and proliferation in HBV-positive liver cancer HCCLM3 cells based on STING signaling and its underlying mechanism.Methods:HCCLM3 cells were treated with quercetin (50 μmol/L or 100 μmol/L), designated as the 50 μmol/L group and 100 μmol/L group, respectively. The inhibitory effect of quercetin on HCCLM3 cells was detected using the CCK-8 method. A scratch assay was conducted to assess the impact of quercetin on the migration ability of HCCLM3 cells. A CCK8 and ROS kit was used to detect the effect of quercetin on the levels of reactive oxygen species in HCCLM3 cells. Western blotting was employed to measure the effect of quercetin on the expression of STING signaling and autophagy-related proteins in HCCLM3 cells. RNA interference technology was used to assess the effects of STING signaling inhibition on the expression of autophagy-related proteins and reactive oxygen species levels in HCCLM3 cells. The combined effects of STING activators and quercetin on HCCLM3 cell proliferation and autophagy were evaluated. The t-test was used to detect data differences between two groups, while ANOVA was employed for comparisons among multiple groups, followed by the SNK- q test for further pairwise comparisons. Results:Compared with the control group, quercetin (50 μmol/L and 100 μmol/L groups) significantly inhibited HCCLM3 cell survival activity in a dose-dependent manner (control group: 100%; 50 μmol/L group: 75.25%; 100 μmol/L group: 50.36%, P<0.01 ). Quercetin inhibited HCCLM3 cell migration in a dose-dependent manner (>2 h, control group: 187.16 μm; 50 μmol/L group: 145.22 μm; 100 μmol/L group: 88.21 μm, P<0.01), which significantly increased intracellular reactive oxygen species (ROS) levels in HCCLM3 cells (control group: 1.00; 50 μmol/L group: 1.565; 100 μmol/L group: 2.175, P<0.01). The phosphorylation level of STING was significantly increased ( P<0.01), and the expression of autophagy-related protein microtubule-related protein 1A/1B light chain 3 (LC3) protein was significantly promoted ( P<0.01). Compared with the quercetin group, the cell viability of the small interfering-STING+quercetin group was increased (quercetin group: 56.3%; small interfering-STING+quercetin group: 85.7%, P<0.05), while the expression of autophagy-related protein LC3 was decreased. Compared with the quercetin group, the cell viability of the quercetin+STING activator group was further decreased (quercetin group: 56.7%; quercetin+STING activator group: 35.4%, P<0.01), and the expression levels of STING and autophagy protein LC3 were significantly increased ( P<0.05). Conclusions:STING signaling-regulated cell autophagy mediates the inhibitory effect of quercetin on the proliferation of HCCLM3 cells, and this effect is enhanced after administration of the STING agonist.

Objective To observe the development and progression of autophagy,and investigate the effect of autophagy on recovery of neurological dysfunction in rats after cerebral ischemia-reperfusion injury.Methods Preparation of middle cerebral artery occlusion (MCAO) models was performed by Longa method.(1) Forty-two SD rats were randomly assigned to blank control 1 group (n=9) and MCAO 1 group (n=33),and the rats of the MCAO 1 group were randomly divided into 6,12,48 and 72 h subgroups (n=6) and 24 h subgroup (n=9) according to the reperfusion times;the ultrastructural changes and autophagosome formation in hippocarnpal tissues of the blank control 1 group (n=3) and 24-h-reperfusion MCAO 1 subgroup (n=3) were observed under transmission electron microscope;the expressions of microtubule associated proteins light chain-3 (LC3)-Ⅱ,LC3-Ⅰ and Beclin-1 in the hippocampal tissues of each group (n=6) were detected by Western blotting.(2) Eighteen SD rats were randomly divided into blank control 2 group,MCAO 2 group and 3-methyladenine (3-MA,autophagy inhibitor) group (60 min prior to MCAO,injection of 10 μL [600 nmoL] 3-MA dilution into the lateral ventricle by stereotactic technique,n=6);the neurological rehabilitation of rats was analyzed by modified neurological severity scale (mNSS) one,3,5 and 7 d after reperfusion;the morphological changes and number of apoptotic cells in the hippocampal tissues were observed by HE staining 7 d after reperfusion.Results (1) The formation ofautophagy in the 24-h-reperfusion MCAO 1 subgroup was clearly observed under microscope;as compared with blank control 1 group,the ratio of LC3-Ⅱ/Ⅰ (excepted for 72-h-reperfusion MCAO 1 subgroup) and Beclin-1 expression in the hippocampus of 6,12,24 and 48-h-reperfusion MCAO 1 subgroups were significantly increased (P＜0.05);as compared with those in the 24-h-reperfusion MCAO 1 subgroup,the ratio of LC3-Ⅱ/Ⅰ and Beclin-1 expression in the hippocampus of 6,12,48 and 72-h-reperfusion MCAO 1 subgroups were significantly decreased (P＜ 0.05).(2) As compared with those the MCAO 2 group,the mNSS scores of 3-MA group were significantly decreased 3,5 and 7 d after surgery (P＜0.05);HE staining indicated that the injury of neurons in the hippocampus of 3-MA group was alleviated,and the number of apoptotic cells in the 3-MA group was significantly smaller than that in the MCAO 2 group ([14.00±2.10]/field vs.[37.83± 2.64]/field,P＜0.05).Conclusion Cerebral ischemia-reperfusion injury can activate autophagy,by which it can alleviate brain damage and improve its neurological dysfunction in rats after cerebral ischemia-reperfusion injury.

Objective To explore the theoretical framework and method of community-based rehabilitation assessment for children with cerebral palsy based International Classification of Functioning, Disability and Health (ICF). Methods The system of assessment of community-based rehabilitation was developed based on the theoretical framework ICF, combined with multi-tools and self-compiled work scales.The system was used for 50 rural children with cerebral palsy after 1-year of community-based rehabilitation. Results and Conclusion The system of assessment is applicable for rural community-based rehabilitation of children with cerebral palsy, with satisfactory efficacy and compliance.

Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146,Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1:6 400, 1:128 00 and 1:128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% (Cpn0146) , 94.5% (Cpn0147) and 96.7% (Cpn0308), respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 (38.8% positive recognition rate), 75 (40.9%), and 82 (44.8%) samples, respectively. The recombinant antigen-based detection assays displayed > 97% of detection specificity and>87%of sensitivity.Condusion GST-Cpn0308 shows a better sensitivity and specificity,which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.

Objective To investigate the attentional bias for emotionally faces in subjects with low-trait anxiety when they are under normal or induce-fear emotion.Methods Trait anxiety inquiry(T-AI) were used to evaluate the mental health status of 480 freshman in a university,and 450 valid questionnaires were obtained.Among them,90 students were identified as low trait anxiety.Sixty students from them were randomized into 2 group(n=30) to receive a dot-probe task with emotional face as stimulus.They were asked to judge the target.The attentional bias under normal or fear-induced emotion to the emotionally faces was examined.Results Low trait anxious college students showed attentional avoidance to the negative faces when they were under normal emotion [D=(-11.09?25.40),t=-2.391,P=0.024],and the orientation was significant [OI=(-20.28?22.90),t=-4.850,P0.05].Conclusion Low trait anxious college students have attentional avoidance to negative faces because of orientation,but the avoidance will disappear when they are under fear emotion.