Objective:To explore the effect of heme-binding protein 1(HEBP1)down-regulation on the function of microglia BV2,and to clarify the key role of HEBP1 in the microglia.Methods:Negative control and HEBP1 knockdown small interfering RNA(siRNA)were constructed to knockdown HEBP1 gene in mouse-derived microglial BV2,and the HEBP1 knockdown BV2 cell models were obtained.The BV2 cells were divided into si-NC group,si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of HEBP1 mRNA and protein in the BV2 cells after knockdown;the siRNA with the most significart knockdown effect was selected for stlbsequent expreriments.The proliferation abilities of the cells in si-NC group and si-HEBP1 group were detected by cell counting kit-8(CCK-8)assay,and the cell migration rates were assessed by scratch assay;the cellular mitochondrial membrane potential and reactive oxygen species(ROS)levels were detected by kits;the cellular mitochondrial respiratory function was detected by mitochondrial respirometer.The BV2 cells were divided into si-NC group,si-NC+lipopolysacch aride(LPS)group,si-HEBP1 group,and si-HEBP1+LPS group.RT-qPCR method was used to detect the expression levels of HEBP1,interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the BV2 cells in various groups,and Western blotting method was used to detect the expression levels of HEBP1 protein in the BV2 cells in various groups.Results:When the BV2 cells were transfected with siRNA carrying with red fluorescence tag CY3,the transfect effricacy was above 90％;compared with si-NC group,the expression levels of HEBP1 protein in the BV2 cells in si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group were significantly decreased(P<0.05 or P<0.01),especially in si-HEBP1-1 group.Compared with si-NC group,the expression levels of HEBP1 mRNA in the BV2 cells in si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group were significantly decreased(P<0.01),especially in si-HEBP1-1 group;indicating that si-HEBP1-1 was the siRNA with best HEBP1 knowdown effect,and the HEBP1 knockdown BV2 cell model was successfully constructed.The CCK-8 resuts showed that compared with si-NC group,the proliferation activities of the BV2 cells in si-HEBP1 group were decreased(P<0.05 or P<0.01);from 90 min,the differences in proliferation activities of the BV2 cells in two groups were obvious.The cell scratch experiment results showed that compared with si-NC group,the cell migration rate in si-HEBP1 group was significantly decreased(P<0.05).The fluorescence microscope results showed that compared with si-NC group,the mitochondrial membrane potential of the BV2 cells in si-HEBP1 group was significantly decreased(P<0.05);compared with si-NC group,the ROS level in the BV2 cells in si-HEBP1 group was significantly increased(P<0.05).The mitochondrial respiration function testing results showed that compared with si-NC group,routine respiration(ROUNTINE)and leak respiration(LEAK)in si-HEBP1 group were significautly decreased(P<0.05 or P<0.01),and electron transfer system capacity(ETS)and residual oxygen consumption(ROX)had no significant differences(P>0.05);the ATP amount was decreased(P<0.05).The RT-qPCR results showed that compared with si-NC group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-NC+LPS group were significantly decreased(P<0.01);compared with si-HEBP1 group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly decreased(P<0.01);compared with si-NC+LPS group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly increased(P<0.01).Conclusion:Knockdown of HEBP1 gene can decrease the proliferation and migration abilities of the microglia BV2 and increase inflammatory response to LPS stimulus,and their mechanisms may be related to mitochondrial function damage and decreased ATP production of the BV2 cells.

Objective Evaluation of Chinese-thyroid imaging reporting and data system(C-TIRADS)combined with contrast-enhanced ultrasound(CEUS)for the assessment of category 4 nodules in the setting of Hashimoto's thyroiditis.Methods Retrospective analysis of 120 C-TIRADS category 4 thyroid nodules from 79 patients with confirmed Hashimoto's thyroiditis who attended the Yiyang Central Hospital from June to December 2022.Thyroid nodules exhibiting one or more benign or malignant features that were suspicious on CEUS were treated as downgraded or upgraded one level.Using the final surgical pathology results as the gold standard,working characteristic(ROC)curves of subjects based on C-TIRADS grading before and after CEUS adjustment were plotted to compare diagnostic efficacy.Results The sensitivity,specificity,and accuracy of the CEUS-adjusted C-TIRADS were 93.0％,87.8％and 90.8％,respectively(P<0.05).The area under the ROC curve was 0.811 and 0.904,respectively(P<0.05).Conclusion C-TIRADS combined with CEUS has better diagnostic efficacy in evaluating category 4 nodules in Hashimoto's thyroiditis.

Objective To find out the social competence level of school-age children with speech sound disorders after cleft palate operation and functional speech sound disorders.Methods Thirty-four school-age children with postoperative cleft palate speech sound disorders and thirty-seven school-age children with functional speech sound disorders attending a specialty clinic for the diagnosis and treatment of childhood speech and language disorders in 2023 were selected,and 32 age-and gender-matched normal children in a local elementary school were also randomly selected as the normal group.The Achenbach Child Behavior Checklist was used to assess the so-cial competence of the three groups of children and a cross-sectional study was conducted.Results Children with cleft palate speech sound disorders and functional speech sound disorders had lower scores on activity,communication and academic ability,with specific manifestations varying in different gender groups,but there was no significant difference in scores between the two groups.In contrast,the activity and communication scores of the functional speech sound disordergroup were significantly lower than those of the normal group(P＜0.05).Conclusion School-age children with cleft palate speech sound disorders and functional speech sound disorders are at a higher risk of difficulties in social functioning,in terms of activity,communication and academic ability,whereas speech sound disorders may be one of the most important influencing factors of such difficulties and cleft palate does not have an additional impact on the social competence level of the child.

Objective To investigate the effects of dihydropteridine reductase (QDPR) on regulating apoptosis induced by plamitic acid(PA). Methods The transfection of HEK293T cells experiment was divided into 3 groups. A group was the control vector group. B group was the control vector group induced by PA. C group was the recombinant plasmid QDPR group induced by PA. First, control vector and recombinant plasmid QDPR was respectively transfected into HEK293T cells. After 24 h, PA with concentration of 0. 5 mmol/L was added into the medium of above cells. The cells of control vector group, the cells of control vector group induced by PA and the cells of recombinant plasmid QDPR group induced by PA were cultured for another 24 hours. At last, cells were harvested to detect tetrahydrobiopterin (BH4) and reactive oxygen species(ROS) generation, Beclin 1, Caspase 3 and Beclin 2 expression. Results ① After transfection, the recombinant plasmid QDPR was successfully constructed and expressed in cells.② There was no significant difference between A group and B group in BH4 generation. Compared with B group, BH4 generation increased in C group (P < 0. 05).③ ROS generation was increased in B group compared with A group, and decreased ROS generation in C group compared with B group (P < 0. 05).④ Western blot analysis revealed that Beclin 1 and Caspase 3 increased (P < 0. 05 ) while Beclin 2 decreased in B group compared with A group (P < 0. 05). Compared to B group, Beclin 1 and Caspase 3 decreased while Beclin 2 increased in C group (P < 0. 05 ). Conclusion QDPR may regulate apoptosis induced by fatty acids by decreasing the generation of ROS and increasing the level of BH4 and the expression of Beclin 2 associated with anti-apoptosis.

Aim To explore the effect and mechanism of resveratrol (Res) on proliferation and differentiation of murine 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were cultured and treated with resveratrol in different dosages.Cell proliferation was analyzed by WST-1 method. Oil red O staining method and spectrophotography were applied to analyze the degree of differentiation. Real-time PCR was applied to detect the mRNA expression of adiponectin and leptin. Western blot was applied to detect the expression of silent information regulator 1 (Sirt1),peroxisome proliferator activated receptor γ (PPARγ) and CCTTA enhancer binding proteinα (C/EBPα).Results Res inhibited proliferation of murine 3T3-L1 preadipocytes in a time-dose dependent manner.The expression levels of adiponectin and leptin mRNA were decreased, and Res also inhibited 3T3-L1 preadipocytes to differentiate into mature adipocytes. Res increased the expression levels of Sirt1 and decreased the expression levels of PPARγ and C/EBPα.Conclusions Resveratrol can inhibit the proliferation and differentiation of 3T3-L1 preadipocytes.The underlying mechanisms may include enhancing expression of Sirt1 and inhibiting expression of PPARγ,C/EBPα which are related to cell differentiation.