Objective:To validate the diagnostic value of soluble scavenger receptor class A (sSRA) in rheumatoid arthritis (RA) and evaluate the clinical utility of enzyme-linked immunosorbent assay (ELISA) and chemiluminescence immunoassay (CLIA) for sSR-A detection.Methods:A total of 200 RA patients (including 37 seronegative RA cases), 19 osteoarthritis (OA) patients, 45 gouty arthritis (GA) patients, and 86 healthy controls (HC) were enrolled from the Department of Rheumatology and Immunology, Northern Jiangsu People′s Hospital, between July 2023 and December 2023. Serum sSR-A levels were measured using ELISA and CLIA, and clinical data were collected. Statistical analyses included t-tests, Mann-Whitney U tests, chi-square tests, Fisher′s exact tests, Kruskal-Wallis tests with Dunn′s post hoc analysis, Spearman correlation, and receiver operating characteristic (ROC) curve analysis. Results:ELISA Detection of sSR-A, RA patients exhibited significantly higher sSR-A levels [0.46(0.25, 0.78) ng/ml] than OA [0.15(0.08, 0.30) ng/ml; Z=4.11, P<0.001], GA [0.22(0.14, 0.44) ng/ml; Z=3.51, P<0.01], and HC [0.15(0.06, 0.24) ng/ml; Z=9.10, P<0.001]. ROC curve analysis revealed that sSR-A had a sensitivity of 68.00% and specificity of 89.41% for diagnosing RA, with an area under the curve (AUC)(95% CI) of 0.834 2(0.787 5, 0.880 8) ( P<0.001), and an optimal cut-off value of 0.291 7 ng/ml. The sSR-A positivity rates in RF/ACPA double-positive RA, RF/ACPA single-negative RA, and seronegative RA subgroups were 74.4% (99/133), 56.7% (17/30), and 54.1% (20/37), respectively, with all subgroups showing significantly higher sSR-A levels than HC ( P<0.001). sSR-A correlated positively with DAS28 ( r=0.60, P<0.001), ESR ( r=0.29, P<0.001), globulin ( r=0.25, P<0.001), IgM ( r=0.26, P=0.044), IgG ( r=0.39, P=0.002), IgA ( r=0.31, P=0.017), RF-IgG ( r=0.47, P<0.001), RF-IgA ( r=0.46, P<0.001), RF-IgM ( r=0.50, P <0.001), and ACPA ( r=0.26, P <0.001). sSR-A levels in high-activity [0.85 (0.65, 1.33) ng/ml], moderate-activity [0.46 (0.27, 0.68) ng/ml], and low-activity RA groups [0.24 (0.11, 0.40) ng/ml] were significantly higher than HC [0.15(0.06, 0.24) ng/ml, Z=7.08, P<0.001; Z=7.45, P<0.001; Z=2.68, P<0.001], with levels increasing with disease activity. RA patients were divided into three groups based on ESR and CRP levels including ESR +CRP + [0.54(0.26, 0.87) ng/ml], ESR/CRP - [0.53(0.25, 0.89) ng/ml], and ESR -/CRP - [0.36(0.18, 0.53) ng/ml], All three groups showed significantly higher sSR-A levels compared to the HC group [0.15(0.06, 0.24) ng/ml, Z=8.02, P<0.001; Z=7.61, P<0.001; Z=5.06, P<0.001]. Moreover, sSR-A levels were significantly positively correlated with grayscale synovitis scores ( r=0.30, P<0.001), power Doppler blood flow scores ( r=0.26, P<0.001), and total semi-quantitative ultrasound scores ( r=0.32, P<0.001), patients who were tested positive for sSR-A exhibited significantly higher grayscale synovitis, power Doppler blood flow, and total ultrasound scores compared to those who were negative for sSR-A ( Z=-2.90, P=0.004; Z=-2.37, P=0.018; Z=-3.09, P=0.002). CLIA detection of sSR-A, the RA group [6.83(3.22, 10.65) ng/ml] demonstrated significantly elevated sSR-A concentrations compared to both the OA+GA [1.99(1.27, 4.18) ng/ml] and HC groups [1.03 (0.50, 1.73) ng/ml] ( Z=4.20, P<0.001; Z=9.62, P<0.001). ROC curve analysis revealed AUC (95% CI) of 0.924 6(0.787 5, 0.880 8), with a sensitivity of 86.92% and a specificity of 93.10% for the diagnosis of RA. Conclusion:sSR-A is a reliable biomarker for diagnosing RA, particularly in seronegative patients, and is closely associated with disease activity. CLIA offers superior diagnostic performance over ELISA in detecting sSR-A in RA patients.

Objective:To evaluate the effect of thunder-fire moxibustion combined with fire-dragon cupping on patients with acute peripheral facial paralysis with co-morbid anxiety and depression.Methods:A total of 80 patients with acute peripheral facial paralysis with co-morbid anxiety and depression were selected and divided into two groups according to the random number table method,with 40 patients in each group.In the control group,acupuncture therapy combined with routine care was given;in the observation group,thunder-fire moxibustion and fire-dragon cupping therapy were added.Both groups were treated once a day,6 times a week,with 1 d of rest,for 4 consecutive weeks(a total of 24 treatments).The House-Brackmann(H-B)grading,facial clinimetric evaluation(FaCE),self-rating anxiety scale(SAS),and self-rating depression scale(SDS)were used before and after the intervention to evaluate the facial nerve function,quality of life,mental health,and clinical efficacy of the two groups.Results:After treatment,patients in both groups showed improvement in H-B grading,scores of all dimensions of the FaCE scale,and SAS and SDS scores compared to those before treatment,and the differences within the groups were statistically significant(P<0.05).After treatment,the total effective rate was 95.0%in the observation group and 75.0%in the control group,and the difference between the two groups was statistically significant(P<0.05).After treatment,the H-B grading of the observation group was better than that of the control group(P<0.05);the scores of all dimensions of the FaCE scale and SAS and SDS scores of the observation group were better than those of the control group(P<0.05).Conclusion:Adding thunder-fire moxibustion and fire-dragon cupping therapy to acupuncture and routine care can promote the recovery of facial nerve function,reduce anxiety and depression,and improve the patients'living standard,and thus is suitable for popularization and application.

Aim To investigate the anti-Zika virus(ZIKV)mechanism of diterpenoid compound 9 from Euphorbia helioscopia in vitro.Methods The cytotox-icity of compound 9 was evaluated using the CCK-8 as-say.A ZIKV-infected Vero cell model was established,and the antiviral activity was assessed through RT-qPCR,plaque assay,Western blot,and immunofluores-cence.Furthermore,the mechanism of action was elu-cidated using multi-cell line validation,nanoparticle tracking analysis,cellular thermal shift assay,and mo-lecular docking.Results In Vero cells,compound 9 exhibited an EC50 of(3.95±0.15)μmol·L-1 and a CC50 of(272.12±8.56)μmol·L-1,demonstrating significantly higher antiviral efficacy than the positive control drug ribavirin(RBV).Its virus inactivation effect was time-dependent and could significantly re-duce viral load and plaque formation.Studies revealed that compound 9 altered the physicochemical properties of ZIKV particles,including reducing surface charge and increasing particle size distribution.Additionally,it significantly enhanced the thermal stability of the prM protein.Molecular docking analysis indicated that compound 9 formed a high-affinity interaction with the prM protein(binding energy:-38.52 kJ·mol-1)and stabilized its structure through hydrophobic interac-tions.Conclusion Compound 9 exerts in vitro anti-ZIKV activity by directly inactivating the virus,disrup-ting viral particle integrity,and targeting the prM pro-tein.

Objective:To analyze and summarize clinical phenotypic characteristics and genetic variations in patients with intellectual disability and pathogenic variations of the DYNC1H1 gene across 4 generations within a single family. Methods:Retrospective case analysis.Clinical data of a child with epilepsy and intellectual disability and her family members were collected from the Children′s Medical Center, Peking University First Hospital on December 2019.The child was followed up regularly.DNA was extracted from the peripheral blood of the child′s family members.Then whole-exome sequencing and Sanger sequencing were performed to identify the genetic variation type in the proband and her family members.The relationship between genotype and phenotype was further analyzed.Results:A total of 13 patients across 4 generations in the family had intellectual disability, and the proband also had drug-resistant epilepsy.The variation c. 13556C> A (p.A4519E) of the DYNC1H1 gene was confirmed by gene testing in 8 patients (no blood samples were obtained from the remaining patients). Conclusions:DYNC1H1 gene-related intellectual disability in most previously reported cases are caused by novel variations of this gene.In this study, a large family of 13 intellectual disability patients across 4 generations caused by a pathogenic mutation in the DYNC1H1 gene was summarized.The findings make precise genetic counseling possible for this family and provide a basis for further studies on the relationship between the genotype and phenotype of the DYNC1H1 gene.

Objective:To evaluate the effect of thunder-fire moxibustion combined with fire-dragon cupping on patients with acute peripheral facial paralysis with co-morbid anxiety and depression.Methods:A total of 80 patients with acute peripheral facial paralysis with co-morbid anxiety and depression were selected and divided into two groups according to the random number table method,with 40 patients in each group.In the control group,acupuncture therapy combined with routine care was given;in the observation group,thunder-fire moxibustion and fire-dragon cupping therapy were added.Both groups were treated once a day,6 times a week,with 1 d of rest,for 4 consecutive weeks(a total of 24 treatments).The House-Brackmann(H-B)grading,facial clinimetric evaluation(FaCE),self-rating anxiety scale(SAS),and self-rating depression scale(SDS)were used before and after the intervention to evaluate the facial nerve function,quality of life,mental health,and clinical efficacy of the two groups.Results:After treatment,patients in both groups showed improvement in H-B grading,scores of all dimensions of the FaCE scale,and SAS and SDS scores compared to those before treatment,and the differences within the groups were statistically significant(P<0.05).After treatment,the total effective rate was 95.0%in the observation group and 75.0%in the control group,and the difference between the two groups was statistically significant(P<0.05).After treatment,the H-B grading of the observation group was better than that of the control group(P<0.05);the scores of all dimensions of the FaCE scale and SAS and SDS scores of the observation group were better than those of the control group(P<0.05).Conclusion:Adding thunder-fire moxibustion and fire-dragon cupping therapy to acupuncture and routine care can promote the recovery of facial nerve function,reduce anxiety and depression,and improve the patients'living standard,and thus is suitable for popularization and application.

OBJECTIVE To evaluate the efficacy and safety of Weiyan Tongluo Granules in treating chronic atrophic gastritis(CAG)and explore its potential mechanisms.METHODS From June 2020 to December 2022,100 CAG patients with spleen defi-ciency and blood stasis syndrome were enrolled and randomly divided into a trial group(n=50)and a control group(n=50)using a random number table.The trial group received Weiyan Tongluo Granules plus a folic acid placebo,while the control group received fo-lic acid tablets plus a traditional Chinese medicine granule placebo.The treatment course for both groups was 24 weeks,with 8 and 10 dropouts in the trial and control groups,respectively.Post-treatment comparisons included OLGA/OLGIM staging reversal rates,low-grade intraepithelial neoplasia regression rate,SSDPRO-CG(Patient-Reported Outcome Scale for Chronic Gastritis in Spleen-Stomach Diseases)scores,TCM syndrome scores,and safety indicators.Serum levels of PG I,PGⅡ,PGR,and G-17 were measured via ELISA before and after treatment.Gastric mucosal p-NF-κB and CDX2 protein expression levels were analyzed by Western blot,while mRNA levels of IL-1β,IL-6,VIL1,and MUC2 were quantified via qPCR.RESULTS After treatment,the trial group showed sig-nificantly higher OLGA and OLGIM stage reversal rates than the control group(P<0.05,P<0.01),though no significant difference was observed in low-grade intraepithelial neoplasia regression.Both groups exhibited significant improvements in physiological domain scores and total SSDPRO-CG scores(P<0.01),with the trial group outperforming the control group in physiological,independence,psychological domains,and total scores(P<0.05,P<0.01).TCM syndrome scores(total and sub-items:gastric distension,pain,poor appetite,bloating)decreased significantly in both groups(P<0.01),while the trial group showed greater reductions in loose stools and dull complexion(P<0.01).After-treatment,the trial group had significantly lower TCM syndrome scores than the control group(P<0.05,P<0.01).Serum PG I,PGⅡ,PGR,G-17,gastric mucosal p-NF-κB,CDX2,and IL-1β,IL-6,VIL1,MUC2 mRNA levels improved significantly in the trial group(P<0.05,P<0.01),while the control group improved only in PGⅡ(P<0.05,P<0.01).The trial group's improvements in these biomarkers surpassed the control group's(P<0.05,P<0.01).No treatment-related adverse events occurred in either group.CONCLUSION Weiyan Tongluo Granules ameliorate gastric mucosal pathology,clinical symptoms,psychological state,and quality of life in CAG patients without significant adverse effects.Its mechanism may involve sup-pressing the NF-κB pathway to reduce IL-1β and IL-6 expression,downregulating CDX2 to inhibit VIL1 and MUC2 transcription,thereby reversing the vicious cycle of inflammation-intestinal metaplasia.

Objective To clarify the differences in the effects of bupivacaine liposome and ropivacaine on quadratus lumborum block in patients undergoing laparoscopic total hysterectomy.Methods A total of 70 patients who underwent laparoscopic total hysterectomy at Suqian First People's Hospital Affiliated to Nanjing Medical University from January 2023 to October 2024 were selected as the research subjects.The patients were randomly divided into the control group and the observation group,with 35 cases in each group.Before general anesthesia,patients of the control group were given ropivacaine quadratus lumborum block,while patients of the observation group were given bupivacaine liposome quadratus lumborum block.The general conditions before and after operation,visual analogue scale(VAS)scores at different time points after operation,rescue analgesia,mini-mental state examination(MMSE)scores at different time points after operation,postoperative rehabilitation and analgesic adverse reactions were compared between the two groups.Results There was no statistically significant difference in the general conditions of patients in the two groups before and after the operation(P>0.05).At 24 hours,48 hours,and 72 hours after surgery,the abdominal resting VAS scores of patients in the observation group were significantly lower than those in the control group(P<0.05).The time of the first self-controlled use of the intravenous analgesic pump after surgery in the observation group was significantly later than that in the control group(P<0.05).The number of analgesic pump compressions and the rate of oxycodone rescue analgesia in the observation group were significantly fewer/lower than those in the control group within 48 hours after surgery(P<0.05).At 24 hours,48 hours,and 72 hours after surgery,the MMSE scores of patients in the observation group were significantly higher than those in the control group(P<0.05).The time for patients to get out of bed and move around after surgery and the recovery time of anal exhaust in the observation group were significantly earlier than those in the control group(P<0.05).There was no statistically significant difference in the incidence of postoperative analgesic adverse reactions between the two groups of patients(P>0.05).Conclusion Compared with the use of ropivacaine for quadratus lumborum block in patients undergoing laparoscopic total hysterectomy,bupivacaine liposome can provide a longer-lasting analgesic effect,reduce patients'dependence on intravenous analgesia,be conducive to protecting patients'cognitive function,accelerate the recovery of patients without increasing the incidence of adverse reactions.

Pasteurella multocida(Pm)type A is an important pathogen responsible for respiratory diseases,such as bovine pneumonia,which seriously restricts the development of cattle industry in China.Currently,the prevention of Pm infection-related diseases primarily relies on vaccination in production.However,the diverse Pm serotypes result in inadequate cross-immunological protection from vaccines.Therefore,it is of great significance to develop vaccines with cross-protection for the prevention and control of Pm infectious diseases.The previous studies conducted by our team have demonstrated that PmCQ2△cra exhibits a strong immune protective effect against Pm type A(PmA),Pm type B(PmB),and Pm type F(PmF).Transcriptomic sequencing results suggest that the cross-immunoprotective effect of PmCQ2△cra may be attributed to high expression levels of bacterial surface protective antigens.Consequently,four putative immune protective antigens,namely PmCQ2-5175,PmCQ2-6290,PmCQ2-0275 and PmCQ2-2640,were screened through bioin-formatics analysis in this study.Subunit vaccines formulated with these potential antigenic proteins exhibited protective efficacy of 62.5％,25％,12.5％and 10％against PmA-infected mice,respectively.Importantly,PmCQ2-5175,one of the most protective single-component antigen vac-cines,demonstrating a 75％cross-protection against PmB infection in mice.Furthermore,the pro-tective efficacy of the PmCQ2-5175 protein screened in this study was superior to that of the previ-ously reported Pm antigen protein plpE.Moreover,the fusion expression protein PmCQ2-5175-PLPE exhibited better protective effects against PmA compared to a single protein.The findings of this study will establish a theoretical foundation for the advancement of Pm subunit vaccines with broad-spectrum immune protection.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective To develop and validate a prediction model for risk of amputation in patients with diabetic foot ulcers(DFU)based on systematic review and meta-analysis.Methods The studies on the risk factors of amputation in DFU patients was retrieved by using subject words+free words.After screening,37 cohort studies were finally included,and the Newcastle-Ottawa scale(NOS)was used for quality evaluation.Meta-analysis was performed on the risk factors of amputation in DFU.Then a prediction model for DFU amputation risk were constructed based on the statistically significant risk factors in the meta-analysis.The corresponding β value was calculated based on the combined odds ratio(OR)value of each risk factor,and each risk factor was scored to establish a scoring system model.The clinical data of 453 DFU patients hospitalized in our department from 2021 to 2023 were collected as a validation cohort.Receiver operating characteristic(ROC)curve analysis was used to evaluate the model performance.The area under the curve(AUC)was calculated,and the optimal cutoff score was determined by calculation of the maximum Youden index through sensitivity and specificity.Results Our meta-analysis showed a cumulative amputation rate of approximately 34.65%in 11 779 DFU patients.The final risk prediction models include gangrene[OR=11.92(5.86～24.24)],ulcer depth[OR=4.93(2.52～9.64)],osteomyelitis[OR=3.19(2.36～4.29)],previous amputation history[OR=3.19(2.00～5.09)]and lower extremity arterial disease[OR=3.10(2.31～4.17)].According to the weights of each risk factor,the total score of the model is 76,and the optimal cut-off score is 36.5.The prediction model performed well,with an AUC value of 0.864(0.824,0.903),a sensitivity of 0.743,a specificity of 0.859,and an accuracy rate of 83.00%.Conclusion A prediction model for DFU amputation risk is developed based on risk factor scoring,and has good discrimination and calibration,providing effective scientific basis for clinical research and clinical decision-making related to DFU amputation.