The ambiguity of etiology makes temporomandibular joint osteoarthritis (TMJOA) "difficult-to-treat". Emerging evidence underscores the therapeutic promise of exosomes in osteoarthritis management. Nonetheless, challenges such as low yields and insignificant efficacy of current exosome therapies necessitate significant advances. Addressing lower strontium (Sr) levels in arthritic synovial microenvironment, we studied the effect of Sr element on exosomes and miRNA selectively loading in synovial mesenchymal stem cells (SMSCs). Here, we developed an optimized system that boosts the yield of SMSC-derived exosomes (SMSC-EXOs) and improves their miRNA profiles with an elevated proportion of beneficial miRNAs, while reducing harmful ones by pretreating SMSCs with Sr. Compared to untreated SMSC-EXOs, Sr-pretreated SMSC-derived exosomes (Sr-SMSC-EXOs) demonstrated superior therapeutic efficacy by mitigating chondrocyte ferroptosis and reducing osteoclast-mediated joint pain in TMJOA. Our results illustrate Alix's crucial role in Sr-triggered miRNA loading, identifying miR-143-3p as a key anti-TMJOA exosomal component. Interestingly, this system is specifically oriented towards synovium-derived stem cells. The insight into trace element-driven, site-specific miRNA selectively loading in SMSC-EXOs proposes a promising therapeutic enhancement strategy for TMJOA.
MicroRNAs/metabolism* ; Mesenchymal Stem Cells/drug effects* ; Osteoarthritis/drug therapy* ; Exosomes/drug effects* ; Strontium/pharmacology* ; Synovial Membrane/cytology* ; Humans ; Animals ; Temporomandibular Joint Disorders/therapy* ; Temporomandibular Joint

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Objective:To evaluate the efficacy and safety of first-line anti-tuberculosis (TB) drugs combined with linezolid in treatment of children with tuberculous meningitis (TBM).Methods:A retrospective cohort study design was performed . Eight-nine Children diagnosed as TBM during January 1 st 2016 and December 31 st 2023 in Department of Infectious Disease, Children′s Hospital of Chongqing Medical University were enrolled in the study. According to different treatment regimens, children were divided into a group of first-line anti-tuberculous drugs (isoniazid, rifampicin, pyrazinamide, ethambutol (HRZE)) and a group of HRZE and linezolid combination (HRZEL). The efficacy and safety of the 2 regimens were compared and the relationship between linezolid drug concentration and adverse reactions were analyzed. Comparisons between groups were performed using χ2 test and Mann-Whitney U test. Results:The 89 children with TBM included 53 males and 36 females with an onset age of 4.6 (1.4, 9.6) years. There were 27 cases in the HZREL group and 62 cases in the HRZE group. Before treatment, positive rate of interferon-gamma release assays (IGRA) in HRZEL group was lower than that in HRZE group (64% (16/25) vs.92% (55/60), χ2=9.82, P<0.05), but protein level of cerebrospinal fluid (CSF) was higher than that in HRZE group (1.2 (1.0, 2.0) vs.0.8 (0.4,1.4) g/L, Z=0.32, P<0.05). By the end of the intensive phase, there were no significant differences of rates of CSF improvement and etiology negativity between HRZEL group and HRZE group (both P>0.05).The 44 TBM children with high CSF protein (>1 g/L) included 25 males and 19 females with an onset age of 6.7 (3.0, 11.8) years. There were 21 cases in the HZREL group and 23 cases in the HRZE group accordingly. Before treatment, there were no significant differences of positive rate of IGRA test and CSF protein level between the 2 groups (62% (13/21) vs. 87% (20/23), 1.7 (1.1, 2.2) vs. 1.5 (1.2, 1.9) g/L, χ2=3.67, Z=0.23, both P>0.05). There were no significant differences in CSF indicators, etiology negativity or imaging remission between the two groups by the end of intensive phase (all P>0.05). Higher frequencies of granulocytopenia, gastrointestinal symptoms as well as withdrawal or change of drugs were found in HRZEL group when compared to those in HRZE group (44% (12/27) vs. 19% (12/62), 7% (2/27) vs. 0, 33% (9/27) vs. 3% (2/62), χ2=6.01, 4.70, 15.74, all P<0.05). Conclusions:The efficacy of HRZEL regimen is similar to conventional HRZE regimen in children with TBM, but with higher adverse effect. Prudentially evaluating the pros and cons of linezolid in the usage of drug-susceptible TB and carefully monitoring of linezolid associated adverse effects is suggested.

ObjectiveTo investigate the association of the polymorphisms of the acetyl-CoA acetyltransferase 1 （ACAT1） gene and the melatonin receptor 1B （MTNR1B） gene with the susceptibility to nonalcoholic fatty liver disease （NAFLD）. MethodsA total of 164 healthy controls and 228 NAFLD patients were enrolled in this study. PCR and sequencing methods were used to determine the genotypes of the polymorphisms of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus， and fasting venous blood samples were collected for biochemical analysis. The t-test was used for comparison of normally distributed continuous data between groups， and the non-parametric Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups； the chi-square test was used for comparison of categorical data between groups. ResultsThere were no significant differences between the NAFLD group and the healthy control group in the genotype distribution of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus （all P>0.05）. The carriers of AA genotype at the rs1044925 locus of the ACAT1 gene had a significantly higher level of low-density lipoprotein than the carriers of C allele （Z=-2.08， P=0.04）， and the carriers of G allele at the rs10830963 locus of the MTNR1B gene had a significantly higher level of fasting blood glucose than the carriers of CC genotype （Z=-3.01， P<0.01）. ConclusionThe polymorphisms of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus were not associated with the susceptibility to NAFLD. The rs1044925 locus of the ACAT1 gene and the rs10830963 locus of the MTNR1B gene are associated with the levels of low-density lipoprotein and fasting blood glucose， respectively.

Objective:Non-alcoholic fatty liver disease(NAFLD)is a common metabolic disorder in overweight and obese children,and its etiology and pathogenesis remain unclear,lacking effective preventive and therapeutic measures.This study aims to explore the association between whole blood copper,zinc,calcium,magnesium and iron levels and NAFLD in overweight and obese children aged 6 to 17 years,providing a scientific basis for the prevention and intervention of early NAFLD in overweight and obese children. Methods:A cross-sectional study design was used to collect relevant data from overweight and obese children who visited the Hunan Children's Hospital from January 2019 to December 2021 through questionnaire surveys.Fasting blood samples were collected from the subjects,and various indicators such as blood glucose,blood lipid,and mineral elements were detected.All children were divided into an overweight group(n=400)and a NAFLD group(n=202).The NAFLD group was divided into 2 subgroups according to the ALT level:A non-alcoholic fatty liver(NAFL)group and a non-alcoholic steatohepatitis(NASH)group.Logistic regression analysis was used to analyze the association between minerals(copper,zinc,calcium,magnesium,and iron)and NAFLD,NAFL and NASH. Results:A total of 602 subjects were included,of whom 73.6%were male,with a median age of 10(9,11)years,and a body mass index(BMI)of 24.9(22.7,27.4)kg/m2.The intergroup comparison results showed that compared with the overweight group,the NAFLD group had higher levels of age,BMI,diastolic blood pressure(DBP),systolic blood pressure(SBP),triglyceride(TG),low density lipoprotein(LDL),alanine transaminase(ALT)and aspartate aminotransferase(AST),and lower level of high density lipoprotein(HDL).The NAFL group had higher levels of age,BMI,DBP,SBP,ALT,and AST,and lower levels of HDL compared with the overweight group.The levels of age,BMI,DBP,SBP,TG,LDL,ALT,and AST of NASH were higher than those in the overweight group,while the level of HDL was lower than that in overweight group(all P<0.017).After adjusting for a variety of confounders,the OR of NAFLD for the highest quantile of iron was 1.79(95%CI 1.07 to 3.00)compared to the lowest quantile,and no significant association was observed between copper,zinc,calcium,and magnesium,and NAFLD.The subgroup analysis of NAFLD showed that the OR for the highest quantile of iron in children with NAFL was 2.21(95%CI 1.26 to 3.88),while no significant association was observed between iron level and NASH.In addition,no significant associations were observed between copper,zinc,calcium,and magnesium levels and NAFL or NASH. Conclusion:High iron level increases the risk of NAFLD(more likely NAFL)in overweight and obese children,while copper,zinc,calcium,magnesium,and other elements are not associated with the risk of NAFLD in overweight and obese children.

BACKGROUND:Modification of food consistency and volume is a commonly used method of swallowing compensation in clinical practice.Dry swallowing is a commonly used method of evaluation.The hyoid muscles are very important in swallowing.The effects of dry swallowing and swallowing tasks of different consistencies and volumes on hyoid muscle activation levels are still unclear. OBJECTIVE:To explore the effects of simple dry swallowing and swallowing tasks of different consistencies and volumes on the hyoid muscles in healthy adults. METHODS:A total of 44 healthy adults were included from April to August 2019,including 19 males and 25 females,with an average age of(21.7±2.8)years.They randomly performed dry swallowing and swallowing tasks of different consistencies(the International Dysphagia Diet Standardisation Initiative(IDDSI)frame levels 0-4)and volumes(5,10,20 mL),and the surface electromyogram signals of the hyoid muscles during each swallowing task were recorded.After processing the raw surface electromyogram signals,the activation levels of the hyoid muscles were compared between dry swallowing and swallowing tasks of different consistency and volume. RESULTS AND CONCLUSION:The mean amplitude values of the suprahyoid muscles corresponding to swallowing tasks of 20 mL for levels 0-4,10 mL for level 3,and 5 mL for level 4 were higher than those of dry swallowing(P＜0.05).The mean amplitude values of the suprahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than those of the 5-mL swallowing tasks of the corresponding consistencies,except for level 3(P＜0.05).The mean amplitude values of the suprahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than those of the 10-mL swallowing tasks of the corresponding consistencies,except for levels 2 and 3(P＜0.05).The mean amplitude values of the infrahyoid muscles corresponding to all swallowing tasks were higher than that of dry swallowing(P＜0.05).The mean amplitude values of the infrahyoid muscles corresponding to the 20-mL swallowing tasks of different consistencies were higher than that of the 5-and 10-mL swallowing tasks of the corresponding consistencies(P＜0.05).The mean amplitude values of the infrahyoid muscles corresponding to the 10-mL swallowing tasks of different consistencies were higher than those of the 5-mL swallowing tasks of the corresponding consistencies,except for level 3(P＜0.05).To conclude,in healthy adults performing swallowing tasks of different volumes and consistencies,the level of activation of the hyoid muscles is less susceptible to IDDSI frame levels 0-4 consistency and more susceptible to volume.The higher volume indicates the higher activation level of the hyoid muscles.

Objective To observe the value of B1 corrected T1 mapping for distinguishing pathological types and differentiation degrees of lung cancers.Methods A total of 74 lesions in 65 patients with lung cancers were prospectively enrolled,including 49 poorly differentiated lesions and 25 moderately or well differentiated ones,i.e.42 adenocarcinomas,14 squamous cell carcinomas and 18 small cell lung cancers(all poorly differentiated).B1 corrected T1 mapping was performed,ROI(ROI1 and ROI2)were delineated using 2 methods,and T1 values of different pathological types and differentiation degrees lung cancers were compared.The receiver operating characteristic(ROC)curves were drawn,and the areas under the curve(AUC)were calculated.Results Significant differences of T1 values were found among different pathological types of lung cancer(all P＜0.05),as well as between small cell lung cancer and the rest 2 types of lung cancer(both P＜0.05).There were significant differences of T1 values between poorly differentiated and moderately well differentiated lung cancer(squamous cell carcinoma+adenocarcinoma)(both P＜0.05).Taken ROI1 T1 value=1 524.21 ms as the cut-off value,the AUC of T1 value for distinguishing poorly differentiated and moderately well differentiated lung cancer(squamous cell carcinoma+adenocarcinoma)was 0.698,with sensitivity of 64.50％and specificity of 76.00％.Taken ROI2 T1 value=1 630.68 ms as the cut-off value,the AUC of T1 value was 0.676,with sensitivity of 54.80％and specificity of 80.00％.Conclusion B1 corrected T1 mapping was helpful for distinguishing pathological types and differentiation degrees of lung cancers.

The universal health coverage agenda promotes population health and social equity and is a priority for the WHO and governments worldwide. This article outlines the basic concept, development, content, monitoring indicators, global progress, and challenges of the universal health coverage agenda. After over half a century of development, a global consensus has been reached on the definition and content of the universal health coverage agenda which emphasizes coverage proportion of the population, content of healthcare services, and economic protection measures. The implementation principle of the agenda for universal health coverage is to prioritize providing healthcare services of high health benefits and social value to the entire population under resource constraints. However, the healthcare service recommendations and evaluation frameworks proposed by the WHO and other international organizations tend to favor low-income countries, neglecting services related to injury prevention and mental health, and therefore may not be suitable for all countries. The development across various dimensions of the agenda for universal health coverage is uneven, with low-income countries lagging. Progress in the prevention and control of non-communicable diseases and injuries is delayed. Low-income groups and vulnerable populations are at a disadvantage in accessing services and economic protection. It is suggested that a globally applicable set of standards, methods, and processes be used to identify high-priority healthcare services. Countries should gradually expand the scope of healthcare services and population coverage based on their needs and capabilities. Additionally, efforts should be made to increase investment in healthcare system resources and international collaboration to promote the development and technological advancement of healthcare systems in low-income countries. Furthermore, it is also necessary to build a high-quality primary healthcare service system and strengthen protection for vulnerable groups.

Objective To investigate the relationship between circular RNA homeodomain interacting protein ki-nase 3 (circHIPK3) and the activation of rat microglia (RM) cells.Methods In vitro, RM cells were cultured and randomized into normal and oxygen-glucose deprivation/reperfusion (OGD/R) groups, and the expression lev-el of circHIPK3 in each group was detected by RT-qPCR.The circHIPK3 lentiviral vector with puromycin resist-ance was constructed, and the overexpression (OE) group and negative control (NC) group were set up.The opti-mal multiplicity of infection (MOI) for RM cells was determined based on fluorescence expression, and puromycin was used to screen RM cells stably expressing circHIPK3 .The cells of OE and NC groups were treated with OGD/R, and the expression levels of ionized calcium binding adaptor molecule 1 (Iba-1) and eukaryotic tumor necrosis factor receptor superfamily (CD40) were detected by Western blot.The circHIPK3 translational protein potential was analyzed by the circRNAdb database, while the potential binding microRNAs on circHIPK3 were predicted by circBank and Starbase databases.Results OGD/R down-regulated circHIPK3 in RM cells (P <0.0001).The sequencing results were accurate and the lentiviral vector of circHIPK3 was constructed successfully.The optimal MOI of RM cells was 80 , puromycin at a concentration of 2μg/ml was used to screen RM cell lines stably express-ing circHIPK3 .RT-qPCR results showed that the expression level of circHIPK3 was significantly higher in the OE group compared with the NC group (P<0.01) .Western blot results revealed that the expression levels of Iba-1 and CD40 in the OE group were markedly lower than those in the NC group (P<0.05) .Protein translation analy-sis showed that circHIPK3 encoded a polypeptide of 404 amino acids with two internal ribosome entry sites (IRES) and an open reading frame (ORF) .Database analysis uncovered that circHIPK3 could target eight specific miR-Nas, namely hsa-miR-3529-5p, hsa-miR-379-5p, hsa-miR-506-3p, hsa-miR-33, hsa-miR-450b-5p, hsa-miR-551b-3p, hsa-miR-193, and hsa-miR-508-3p.Conclusion The overexpression of circHIPK3 effectively suppres-ses OGD/R-induced activation of RM cells.It has the potential to encode peptides and may act as a miRNA sponge.These findings provide a foundation for further study of circHIPK3 functions.