ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.

Objective: The efficacy of antidepressants is influenced by a combination of genetic, individual, and environmental factors. This study aimed to investigate the association between the miR-182 rs76481776 polymorphism and the response to antidepressant treatment in major depressive disorder (MDD) patients, and its underlying molecular mechanisms. Methods: This study enrolled 180 MDD patients and 180 healthy controls. The rs76481776 genotype was determined using TaqMan-based qPCR. The severity of depression and treatment response were assessed using the Hamilton Depression Rating Scale (HAMD). The expression of miR-182 and BDNF was measured using RT-qPCR. The regulatory relationship between miR-182 and BDNF was confirmed through a luciferase reporter gene assay. The correlation between miR-182 and BDNF expression was evaluated using the Pearson correlation coefficient. Results: The T allele of rs76481776 was a significant risk factor for MDD (OR = 2.182, 95% CI: 1.424−3.345, p ＜ 0.001) and was significantly associated with higher baseline scores on the HAMD. Moreover, individuals carrying the T allele (CT/TT genotype) exhibited a significantly poorer response to antidepressant treatment and a lower remission rate within 12 months compared to those with the CC genotype (p ＜ 0.05). Mechanistically, miR-182 was highly expressed in patients with MDD (p ＜ 0.05), and its expression was even higher in T allele carriers (p ＜ 0.05). miR-182 could directly target and suppress BDNF, leading to decreased BDNF expression in MDD patients, and its expression was significantly and inversely correlated with BDNF expression. Conclusion The T allele of rs76481776 diminished the therapeutic efficacy of antidepressants by up-regulating miR-182 expression and subsequently suppressing BDNF expression.

ObjectiveTo prepare a recombinant PETase with a dual-catalytic-triad and to evaluate its efficiency in the biodegradation of polyethylene terephthalate （PET）. MethodsBased on the crystal structure of wild-type PETase， point mutations （T88H/L117D） were introduced via site-directed mutagenesis. The recombinant protein was prepared using prokaryotic expression and chromatography purification techniques. The enzymatic hydrolysis of the mutant PETase was assessed by relatively quantifying the products mono （2-hydroxyethyl） terephthalate （MHET） and terephthalic acid （TPA）. ResultsBoth wild-type and mutant PETases accumulated as inclusion bodies， accounting for approximately 20% of the total bacterial protein. After solubilization in urea， the proteins were eluted at 300 mmol/L imidazole during affinity chromatography purification， with concentrations of 1.824 and 1.833 mg/mL and purities of 83.11% and 84.32%， respectively. Subsequent anion-exchange chromatography yielded highly pure enzymes in the 200 mmol/L NaCl fraction： 2.776 mg/mL （96.86% purity） for the wild type and 1.967 mg/mL （95.13% purity） for the mutant. Following refolding， the final concentrations were 0.484 mg/mL for the wild type and 0.991 mg/mL for the mutant. Hydrolysis assays revealed that the mutant released MHET and TPA at （237.67±17.00）% and （197.33±12.01）% of the wild-type levels， respectively. ConclusionThe T88H/L117D dual-catalytic-triad PETase is successfully prepared and it significantly enhanced PET-degrading activity， thus， it′s a promising biocatalyst for PET bioremediation.

Objective: To analyze the intervention effects of moderate to vigorous classroom physical exercise on psychological sub health among Uyghur junior high school students, providing reference for mental health promotion and intervention in the ethnic group. Methods: From April to July 2022, 647 eighth grade Uighur junior high school students from two middle schools in the Kashgar Prefecture, Xinjiang, were selected using a random cluster digital sampling method. They were randomly assigned to a control group ( n =326) and an intervention group ( n =321). While adhering to the regular curriculum, the intervention group additionally received 12 week, 36 session (every Monday, Wednesday, and Friday afternoon) moderate to vigorous classroom physical exercise intervention (45 minutes per session, including warm up, main exercises, and cool down activities). Self administered psychological sub health was completed through Brief Instrument on Psychological Health of Youths before and after intervention, and the data were analyzed using covariance analysis. Results: Before intervention, there were no statistically significant differences between the control and intervention groups in the total scores for psychological sub health or in the scores for the three dimensions including emotional problems, conduct problems, and social adaptation difficulties ( t =0.76, 0.40, 0.01, 0.94 , all P >0.05). After intervention, the scores of the intervention group s junior high school students on the emotional problems, conduct problems, and social adaptation difficulties dimensions decreased by 0.50, 0.28, and 0.18, respectively, compared with the control group, and all differences were statistically significant ( t =3.35, 3.36, 2.07, all P <0.05). The main effect of the psychological sub health category was statistically significant ( F =60.66, P <0.01, partial η 2=0.09). Compared with the control group, the psychological sub health scores of junior high school students in the intervention group decreased by 0.96 ( t =3.42, P < 0.05 ). Conclusion The 12 week moderate to vigorous classroom physical exercise intervention has a positive effect on the psychological sub health of Uyghur junior high school students.

OBJECTIVE: To observe the clinical efficacy of acupuncture based on "status-target coherence" theory combined with Kegel exercises for vaginal laxity syndrome (VLS). METHODS: Sixty-six patients with VLS were randomized into an observation group (33 cases, 2 cases dropped out, 1 case was discontinued) and a control group (33 cases, 5 cases dropped out). The observation group was treated with acupuncture combined with Kegel exercises, acupuncture was applied to bilateral Ciliao (BL32), Zhongliao (BL33), Sanyinjiao (SP6), etc. The control group was treated with Kegel exercises. Both acupuncture and Kegel exercises were performed once every other day, three times a week for 12 weeks. Before and after treatment, the vaginal laxity questionnaire (VLQ) score, pelvic floor muscle strength (vaginal resting pressure, vaginal systolic pressure, vaginal contraction duration), degree of vaginal laxity and sexual satisfaction questionnaire (SSQ) grade were observed in both groups. RESULTS: After treatment, the VLQ score, vaginal resting pressure, vaginal systolic pressure, vaginal contraction duration in the observation group were elevated compared with those before treatment (P<0.05), and SSQ grade was improved (P<0.05); and the above indexes in the observation group were better than those in the control group (P<0.05). There were no significant difference before and after treatment in the degree of vaginal laxity in the two groups (P>0.05). CONCLUSION Acupuncture based on "status-target coherence" theory combined with Kegel exercises can effectively enhance the strength of pelvic floor muscles, improve the symptoms of vaginal laxity, and improve the satisfaction of sexual life, and its therapeutic effect is better than Kegel exercises alone.
Humans ; Female ; Adult ; Vagina/physiopathology* ; Acupuncture Therapy ; Exercise Therapy ; Young Adult ; Middle Aged ; Treatment Outcome ; Acupuncture Points ; Combined Modality Therapy ; Vaginal Diseases/therapy*

This study employs ultra-performance liquid chromatography(UPLC) to analyze the differences in alkaloid content of Commelina communis from various geographical origins, exploring its feasibility as a quality evaluation indicator. A total of 57 batches of C. communis samples from 23 provinces, autonomous regions, and municipalities in China were selected. The MicroPulite HSS T3(2.1 mm×50 mm, 1.8 μm)column was used with a mobile phase of acetonitrile-0.2% phosphoric acid aqueous solution(20∶80), detection wavelength at 254 nm, and a flow rate of 0.3 mL·min~(-1) to measure the content of 1-deoxynojirimycin(DNJ) and deoxymannojirimycin(DMJ). The MicroPulite XP tC_(18)(2.1 mm×100 mm, 1.7 μm)column was employed with a mobile phase of acetonitrile-0.2% phosphoric acid aqueous solution(4∶96), detection wavelength at 254 nm, and a flow rate of 0.4 mL·min~(-1) to measure the content of norharmine(NHM) and harmanme(HM). Chemometric methods were applied to study the relationships and differences among the 57 batches of C. communis. Significant differences in alkaloid content were observed among C. communis from different regions, with the average total content decreasing in the order of North China, Northeast China, Northwest China, East China, Southwest China, Central China, and South China. Cluster analysis(CA) and principal component analysis(PCA) further revealed the quality differences of C. communis from various origins, and partial least squares discriminant analysis(PLS-DA) identified DNJ as a marker compound to distinguish the quality differences between different geographical sources of C. communis. It is recommended that the content limit of DNJ be set at no less than 0.055 9%, providing a reference for the quality evaluation and clinical application of C. communis medicinal materials.
Alkaloids/analysis* ; Drugs, Chinese Herbal/chemistry* ; China ; Chromatography, High Pressure Liquid ; Chemometrics/methods* ; Quality Control

Objective:To investigate the application of time series models in forecasting pre-hospital emergency ambulance trips in Handan City and develop the LPro ensemble model for improved prediction accuracy to support emergency resource allocation.Methods:Pre-hospital emergency data from Handan Emergency Medical Command Center (2019-2023) were retrospectively analyzed. From 324 799 original records, 289 949 valid records were included after cleaning. The training set (2019-2022: 215 918 records) included 35 527 records in 2019, 52 015 in 2020, 61 836 in 2021, and 66 540 in 2022. The validation set (2023) contained 74 031 records. ARIMA, linear trend seasonal, exponential smoothing, and Prophet models were fitted to the training set. The LPro ensemble model was constructed using MAPE-based weighting (linear trend seasonal model: 0.38, Prophet: 0.62). Performance metrics included MAPE, RMSE, MAE, and R 2. Results:Data showed annual growth (compound annual growth rate 23.27%) and seasonal patterns (October peaks, February troughs). Ambulance dispatches increased annually with monthly cyclical patterns. For 2023 validation predictions: ARIMA (MAPE 8.76%, RMSE 619, MAE 491, R 2 0.4563), linear trend seasonal (MAPE 9.83%, RMSE 671, MAE 545, R 2 0.3608), Prophet (MAPE 8.43%, RMSE 562, MAE 503, R 2 0.5513), exponential smoothing (MAPE 8.08%, RMSE 643, MAE 410, R 2 0.4124). LPro model showed superior performance (MAPE 7.05%, RMSE 491, MAE 393, R 2 0.6570), with 16.37% lower MAPE, 12.63% lower RMSE, 21.87% lower MAE, and 19.17% higher R 2 versus Prophet. Conclusion:The LPro ensemble model substantially enhances prediction accuracy and reliability, offering scientific support for emergency resource optimization and dispatch scheduling in Handan City.

Ankylosing spondylitis(AS)is a chronic autoimmune disease characterized by inflammatory involvement of the axial skeleton and pathological bone formation.The T helper 17 cell(Th17 cell)subset of lym-phocytes plays a central role in mediating the inflammatory processes associated with AS.This review summarizes recent advances in the regulation of Th17 cell differentiation in AS,with a focus on the complex mechanisms governed by cytokine microenvironments,transcription factor networks,and metabolic and epigenetic regulatory pathways.Key regulatory components discussed include the IL-23/STAT3 signaling axis,the CCL20/CCR6 chemo-tactic axis,and the master transcription factor RORγt.Additionally,this review critically evaluates emerging thera-peutic strategies targeting metabolic reprogramming(e.g.,PKM2),epigenetic regulators(e.g.,JMJD3,EZH2),engineered exosome delivery systems,and modulators of metabolic enzymes.By analyzing the limitations of current treatment approaches,the review proposes future research directions emphasizing multi-target therapeutic strategies and highlights the importance of personalized medicine in achieving precise and effective treatment for AS.These developments reveal promising new avenues for modulating Th17-mediated immunity,offering transformative poten-tial for the clinical management of AS.

The excessive accumulation of advanced glycosylation end products(AGEs), the end products of non-enzymatic glycosylation reactions, can be involved in the pathological processes of various ocular diseases through mechanisms such as oxidative stress, inflammatory responses and apoptosis. In this paper, we systematically reviewed the key role of AGEs in diabetic keratopathy, cataract, glaucoma, age-related macular degeneration(ARMD)and diabetic retinopathy(DR). It was found that AGEs activate signalling pathways such as NADPH oxidase, MAPK and NF-κB by binding to the receptor RAGE, leading to reactive oxygen species(ROS)generation, release of inflammatory factors, and vascular endothelial dysfunction, which in turn induces delayed corneal healing, cross-linking of lens proteins, optic nerve degeneration, formation of choroidal neovascularisation(CNV), and blood-retinal barrier(BRB)disruption. For example, in diabetic keratopathy, AGEs delay wound healing via the ROS/NLRP3 inflammatory vesicle axis; in cataract, ascorbic acid-mediated cross-linking of lens proteins due to AGEs directly impairs lens transparency; and in DR, AGEs exacerbate microvascular damage by regulating vasucular endothelial growth factor(VEGF)expression and pericyte apoptosis. In addition, this article discusses the advances and limitations of AGEs detection techniques, such as the potential application of lens AGEscan fluorescence assay in screening for diabetic complications, and the need to develop tissue-specific assays for aqueous humour and vitreous. For therapeutic strategies, the research directions of inhibiting AGEs production, blocking RAGE signalling pathway and developing anti-glycosylation drugs are proposed to emphasise their clinical value in delaying disease progression. This review not only integrates the molecular mechanisms and clinical associations of AGEs in ocular diseases, but also provides a theoretical basis for targeted interventions, which is of great significance in exploring novel diagnostic and therapeutic strategies.

Objective To investigate the efficacy and influencing factors of programmed death-1(PD-1)inhibitors in neoadjuvant chemotherapy for triple-negative breast cancer(TNBC).Methods A total of 86 patients with TNBC who received neoadjuvant therapy in The Fifth Medical Center,PLA General Hospital between Jan.1,2018,and Jan.1,2024 and met the inclusion criteria were enrolled,and their clinicopathological data were collected.Based on the neoadjuvant treatment regimens,40 patients who received TP+PD-1 inhibitor(paclitaxel+carboplatin+pembrolizumab)were assigned to TP+PD-1 inhibitor group,and 46 patients who received TP(paclitaxel+carboplatin)were assigned to TP group.The efficacy and incidence of adverse events were compared between the 2 groups after 6 cycles of neoadjuvant therapy.According to the efficacy of neoadjuvant therapy,the patients were further categorized into pathological complete response(pCR)group and non-pCR group.Multivariate logistic stepwise regression analysis was performed to identify independent factors influencing neoadjuvant treatment efficacy.Patients were followed up until Dec.31,2024,and survival analysis was conducted using Kaplan-Meier method.Results There was no significant difference in the objective response rates between the TP+PD-1 inhibitor group and TP group after neoadjuvant therapy(95.0%[38/40]vs 91.3%[42/46],P=0.351].However,the pCR rate was significantly higher in the TP+PD-1 inhibitor group compared with the TP group(65.0%[26/40]vs 43.5%[20/46],P=0.047).There were no significant differences between the 2 groups in terms of disease-free survival,overall survival,or incidence of adverse events(all P＞0.05).Multivariate logistic stepwise regression analysis revealed that the expression of Ki-67 and treatment regimen were influencing factors of pCR after neoadjuvant therapy(odds ratio[OR]=3.382,95%confidence interval[95%CI]1.290-8.868,P=0.013;OR=2.524,95%CI 1.013-6.285,P=0.047).One case of distant metastasis and death occurred in the pCR group,while 8 cases of distant metastasis and 4 deaths occurred in the non-pCR group.The disease-free survival was significantly longer in the pCR group than in the non-pCR group(P=0.031),while the overall survival was similar between the 2 groups(P=0.087).Conclusion Compared with the 6-cycle TP regimen,the 6-cycle TP combined with PD-1 inhibitor regimen can improve the pCR rate in the neoadjuvant treatment of TNBC,with manageable adverse events,suggesting it may serve as a preferred option for TNBC neoadjuvant therapy.Ki-67 expression may serve as a predictive biomarker for achieving pCR.TNBC patients who achieved pCR have better disease-free survival than those who did not.