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.

Nucleic acid aptamers are short oligonucleotide sequences that can specifically bind to target substances with affinity comparable to or even better than protein antibodies.Due to their advantages of low cost,ease of modification,and low immunogenicity,aptamers have shown great potential in early cancer screening,diagnosis and targeted therapy.This paper systematically re-views the latest research progress on nucleic acid aptamers in immunotherapy,gene therapy,and drug delivery,discusses the current status and technical challenges of aptamers in cancer therapy and outlines future research directions.

Nucleic acid aptamers are short oligonucleotide sequences that can specifically bind to target substances with affinity comparable to or even better than protein antibodies.Due to their advantages of low cost,ease of modification,and low immunogenicity,aptamers have shown great potential in early cancer screening,diagnosis and targeted therapy.This paper systematically re-views the latest research progress on nucleic acid aptamers in immunotherapy,gene therapy,and drug delivery,discusses the current status and technical challenges of aptamers in cancer therapy and outlines future research directions.

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 explore the repair effect of nucleus pulposus spherical cell clusters in the process of intervertebral disc degeneration and its potential therapeutic application.Methods:The degradation and rheological properties of SupraGel hydrogel were characterized, and the biocompatibility of spherical nucleus pulposus cell clusters in SupraGel hydrogel was tested by live-dead cytological staining. A caudal disc puncture degeneration model was established in 20 SD rats, and the rats were divided into groups and intervention measures: puncture of degenerated segment without intervention (positive control group), puncture of degenerated segment injection of SupraGel hydrogel (hydrogel group), puncture of degenerated segment injection of spherical cell clusters (cell cluster group), puncture of degenerated segment injection of SupraGel hydrogel loaded with spherical cell clusters (hydrogel + cell cluster group) and healthy segment (negative control group). X-rays and MRI examinations were performed at the 4th and 8th weeks after intervention, and the disc degeneration was evaluated according to the disc height index and Pfirrmann grading. HE staining, safranin O-fast green staining and COL-2 immunohistochemical staining were used to evaluate the changes in the disc structure and extracellular matrix.Results:The degradation residual rate of SupraGel hydrogel was 73.7% after 10 h, which showed excellent shear thinning and self-recovery properties. Nucleus pulposus cells could spontaneously aggregate in SupraGel hydrogel to form stable spherical cell clusters, which remained active on the 7th day. In the in vivo experiment, the intervertebral height index of the hydrogel + cell cluster group was 88.12%±4.55% and 74.13%±4.09% at 4 and 8 weeks after intervention, which were higher than those of the positive control group and hydrogel group, and lower than those of the negative control group, with statistically significant differences ( P<0.05); the Pfirrmann grades were 2.40±0.55 and 2.60±0.89, which were lower than those of the positive control group and hydrogel group at 4 weeks, and lower than those of the positive control group, hydrogel group and cell cluster group at 8 weeks, and higher than those of the negative control group, with statistically significant differences ( P<0.05); the morphological scores were 6.60±0.55 and 9.40±1.14, which were lower than those of the positive control group, hydrogel group and cell cluster group at 4 weeks, at the 8th week, the levels of collagen type II were lower than those of the positive control group, hydrogel group and cell cluster group, but higher than those of the negative control group, and the difference was statistically significant ( P<0.05). The optical density of type II collagen was 0.12±0.01 and 0.10±0.01, respectively, which were higher than those of the positive control group, hydrogel group and cell cluster group at the 4th week, and higher than those of the positive control group and hydrogel group at the 8th week, and the difference was statistically significant ( P<0.05). Conclusion:Nucleus pulposus spherical cell clusters can be successfully constructed and have been shown to have the effect of delaying degeneration in the rat intervertebral disc degeneration model.

Objective:To explore the repair effect of nucleus pulposus spherical cell clusters in the process of intervertebral disc degeneration and its potential therapeutic application.Methods:The degradation and rheological properties of SupraGel hydrogel were characterized, and the biocompatibility of spherical nucleus pulposus cell clusters in SupraGel hydrogel was tested by live-dead cytological staining. A caudal disc puncture degeneration model was established in 20 SD rats, and the rats were divided into groups and intervention measures: puncture of degenerated segment without intervention (positive control group), puncture of degenerated segment injection of SupraGel hydrogel (hydrogel group), puncture of degenerated segment injection of spherical cell clusters (cell cluster group), puncture of degenerated segment injection of SupraGel hydrogel loaded with spherical cell clusters (hydrogel + cell cluster group) and healthy segment (negative control group). X-rays and MRI examinations were performed at the 4th and 8th weeks after intervention, and the disc degeneration was evaluated according to the disc height index and Pfirrmann grading. HE staining, safranin O-fast green staining and COL-2 immunohistochemical staining were used to evaluate the changes in the disc structure and extracellular matrix.Results:The degradation residual rate of SupraGel hydrogel was 73.7% after 10 h, which showed excellent shear thinning and self-recovery properties. Nucleus pulposus cells could spontaneously aggregate in SupraGel hydrogel to form stable spherical cell clusters, which remained active on the 7th day. In the in vivo experiment, the intervertebral height index of the hydrogel + cell cluster group was 88.12%±4.55% and 74.13%±4.09% at 4 and 8 weeks after intervention, which were higher than those of the positive control group and hydrogel group, and lower than those of the negative control group, with statistically significant differences ( P<0.05); the Pfirrmann grades were 2.40±0.55 and 2.60±0.89, which were lower than those of the positive control group and hydrogel group at 4 weeks, and lower than those of the positive control group, hydrogel group and cell cluster group at 8 weeks, and higher than those of the negative control group, with statistically significant differences ( P<0.05); the morphological scores were 6.60±0.55 and 9.40±1.14, which were lower than those of the positive control group, hydrogel group and cell cluster group at 4 weeks, at the 8th week, the levels of collagen type II were lower than those of the positive control group, hydrogel group and cell cluster group, but higher than those of the negative control group, and the difference was statistically significant ( P<0.05). The optical density of type II collagen was 0.12±0.01 and 0.10±0.01, respectively, which were higher than those of the positive control group, hydrogel group and cell cluster group at the 4th week, and higher than those of the positive control group and hydrogel group at the 8th week, and the difference was statistically significant ( P<0.05). Conclusion:Nucleus pulposus spherical cell clusters can be successfully constructed and have been shown to have the effect of delaying degeneration in the rat intervertebral disc degeneration model.

Objective:To analyze the characteristics of patients with Takayasu arteritis (TA) in the east China Takayasu arteritis (ECTA) cohort and their subgroups, and evaluate the disease characteristics.Methods:Patients diagnosed with TA in ECTA cohort from January 2009 to October 2019 were enrolled and their data were analyzed. The characteristics were analyzed and compared within subgroups using t-test or Wilcoxon rank sum test or Chi-square test. Results:A total of 454 patients were included, with the male to female ratio of 1∶4.75(79/375), and the main complaint were dizziness/headache, fatigue, and chest tightness/pain. The type Ⅴ and Ⅰ were the most common angiographic pattern, among which the subclavian artery and carotid artery were most vulnerable, manifested as vascular stenosis. Hypertension, tuberculosis and hepatitis B were common complications. In subgroup comparison, symptoms and inflammation index were much more evident in the active group, female group, <40 years old, and newly diagnosed group. C-reactive protein (CRP)[10(2, 33) mg/L vs 3(1, 14) mg/L, Z=-4.49, P<0.01), erythrocyte sedimentation rate (ESR) [(45±33) mm/1 h vs (25±23) mm/1 h, t=-5.82, P<0.01), in the active group were significantly higher than those in the inactive group, while the ESR in female patients was only higher than that in males, but without statistical significant difference. SAA in the young age group, ESR in the newly diagnosed group was significantly higher than that in the other subgroups [19(6, 95) mg/L vs 10(4, 39) mg/L, Z=2.06, P<0.05] [(44±34) mm/1 h vs (32±28) mm/1 h, t=3.77, P<0.01]. Conclusion:The TA patients are mainly young women, and are in active disease when first being diagnosed. The type Ⅴ and Ⅰ are the most common artery involve-ment pattern. Hypertension and tuberculosis are the most frequent complications.

Objective:To compare the diagnostic efficacy of Chinese diagnostic model, the 1990 American College of Rheumatology (ACR) classification criteria and the 2018 ACR new classification criteria (draft) for Takayasu arteritis (TA).Methods:A total of 196 TA patients who came to our hospital from January 1, 2009 to May 31, 2019 in the TA database of the department of rheumatology and immunology, Zhongshan Hospital, Fudan University and 131 patients with other vascular diseases visited during the same period were selected. General characteristics, clinical data, laboratory tests and imaging tests of all patients were collected. Categorical variables were presented as numbers and percentages, between-group differences were analyzed using the χ2 test. Continuous variables were presented as the Mean± SD for a normal distribution or median and interquartile range (IQR) for a non-normal distribution. Between-group differences were analyzed using the Student's t-test or Mann- Whitney test, as appropriate. The sensitivity, specificity, positive predictive value, negative predictive value, accuracy and receiver operating characteristics of the above diagnostic/classification criteria area under the curve were analyzed. P<0.05 was considered significant. Results:In terms of sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and area under receiver operating characteristic curve (ROC), Chinese diagnostic models was 85.7%, 96.2%, 89.9%, 97.1%, 81.5%, 0.909, 1990 ACR criteria was 47.4%, 97.7%, 67.6%, 96.9%, 55.4%, 0.726, 2018 ACR classification criteria was 79.1%, 98.5%, 86.5%, 98.7%, 75.9%, 0.888. The difference between the Chinese diagnostic model and the 2018 ACR criteria in AUC was not statistically significant ( Z=1.186 , P>0.05). The sensitivity, accuracy and diagnostic efficiency of Chinese diagnostic model was the best, that of the 1990 ACR classification criteria was the worst, and the specificity of the 2018 ACR classification criteria was the highest. The Kappa value of the 2018 ACR classification criteria and the Chinese diagnostic model was 0.719, which had good consistency, and the Kappa value of the consistency between the 1990 ACR classification criteria and the Chinese TA diagnostic model was 0.516. Conclusion:The Chinese diagnostic model, which is based on the clinical characteristics of the Chinese TA population, has a good diagnostic efficacy for the Chinese population. The 2018 ACR classification criteria (draft) is highly consistent with the Chinese TA diagnostic model, and can be promoted and applied in practice.