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.

ObjectivePost-ischemic acute inflammation and the subsequent persistent dysregulation of the immune microenvironment represent major pathological drivers that aggravate neuronal injury and severely restrict functional recovery following ischemic stroke. Although current reperfusion therapies partially restore blood flow, they fail to effectively modulate the secondary inflammatory cascade and oxidative stress, which remain critical barriers to neurological restoration. To address this challenge, this study aimed to engineer and systematically evaluate a biomimetic nanosystem composed of transforming growth factor-β1 (TGF-β1)-loaded platelet membrane-camouflaged lipid nanoparticles (PLP). This nanosystem was designed to achieve dual lesion-targeted delivery and immune microenvironment remodeling. By verifying its spatiotemporal accumulation, anti-inflammatory activity, and neuroprotective efficacy, we sought to establish an integrated therapeutic strategy that simultaneously enables lesion targeting, immune regulation, and functional recovery after ischemic injury. MethodsThe physicochemical properties of PLP, including hydrodynamic particle size, zeta potential, structural stability, and morphology, were characterized using dynamic light scattering, zeta potential analysis, and transmission electron microscopy. The preservation of platelet membrane-derived adhesion and immunoregulatory proteins was confirmed by SDS-PAGE through comparative analysis of protein band profiles between PLP and native platelet membranes. The in vitro biological activities of PLP were evaluated using two complementary cellular models. LPS-induced M1-polarized RAW264.7 macrophages were employed to assess inflammatory modulation, while oxygen glucose deprivation/reperfusion (OGD/R)-induced BV2 microglial cells and SH-SY5Y neuronal cells were utilized to investigate neuroinflammatory regulation and neuronal protection. For in vivo validation, a transient middle cerebral artery occlusion (tMCAO) mouse model was established to mimic ischemia-reperfusion injury. The spatiotemporal biodistribution and lesion-targeting capability of the PLP were monitored through live fluorescence imaging. Therapeutic efficacy was comprehensively evaluated by triphenyltetrazolium chloride (TTC) staining, glial fibrillary acidic protein (GFAP) immunofluorescence analysis, body weight monitoring, and neurological severity score (NSS) assessment. ResultsPLP nanoparticles displayed a uniform spherical morphology, nanoscale particle size distribution, and stable negative surface charge, indicating favorable colloidal stability and circulation potential. SDS-PAGE results confirmed the effective retention of key platelet membrane proteins associated with endothelial adhesion, immune evasion, and inflammatory regulation, demonstrating the successful biomimetic construction. Optimal therapeutic concentrations were determined in OGD/R-induced BV2 cells, where PLP exhibited excellent cytocompatibility and anti-inflammatory activity.In vitro experiments demonstrated that PLP significantly inhibited the polarization of RAW264.7 macrophages toward the pro-inflammatory M1 phenotype and markedly reduced neuronal apoptosis under ischemia-reperfusion conditions. In vivo fluorescence imaging revealed that PLP rapidly accumulated in the ischemic brain hemisphere and maintained prolonged retention for up to 7 d, suggesting enhanced lesion-specific targeting and sustained drug release. Compared with control group, PLP treatment significantly reduced cerebral infarct volume, attenuated reactive astrogliosis, improved weight recovery, and accelerated neurological functional restoration, as reflected by significantly improved NSS scores. ConclusionThis study establishes a multifunctional biomimetic nanoplatform that integrates platelet membrane-mediated active targeting with the anti-inflammatory, antioxidative, and neuroprotective properties of TGF-β1. The PLP system enables rapid lesion homing and long-term retention while synergistically regulating the post-stroke inflammatory microenvironment by suppressing pro-inflammatory immune activation, reducing neuronal apoptosis, and limiting excessive astrocyte reactivity. Importantly, this study proposes a conceptually therapeutic paradigm that combines targeted delivery with immune microenvironment remodeling to achieve comprehensive neurovascular protection. These findings provide strong experimental evidence supporting the translational potential of biomimetic nanotherapeutics as next-generation precision interventions for ischemic stroke.

bjective To analyze the growth trends of height and weight among primary and secondary school students, and explore the developmental characteristics and gender differences at different age groups, and to provide a scientific basis for adolescent health policy formulation. Methods Based on 675 175 health examination records of 227 978 students aged 6-17 years in Shiyan City from 2015 to 2024, a logistic growth model was employed to fit the curves of height and weight changes with age. Results From 2015 to 2024, height and weight showed steady increases across all age groups, exhibiting typical sigmoidal growth patterns. The growth rates varied across age groups: the younger age group (6-9 years) showed a moderate growth (annual height increase of 0.5-1.0 cm, weight increase of 0.03-0.06 kg/year), while the older age group (10-17 years) demonstrated a significant growth (annual height increase of 1.5-2.0 cm, weight increase of 0.22-0.38 kg/year). The growth rate curves displayed a unimodal distribution. The growth inflection points of male students occurred later than that of female students (height inflection point: 9.87 years for males vs. 8.98 years for females; weight inflection point: 10.70 years for males vs. 9.99 years for females). Female students experienced a more concentrated but shorter period of growth and development. The peak height growth rate was 7.40 cm/year at age 9 for females and 7.09 cm/year at age 10 for males, while the peak weight growth rate was 5.04 kg/year at age 10 for females and 5.27 kg/year at age 11 for males. Conclusion The physical development of primary and secondary school students in Shiyan City follows a logistic growth pattern, with significant gender differences and characteristics of adolescent growth spurts. Female students exhibit an earlier and more concentrated growth process.

ObjectiveTo develop and preliminarily validate a prediction model for brace treatment failure risk in patients with adolescent idiopathic scoliosis (AIS), based on information obtained before brace treatment and during initial brace fitting. MethodsA total of 191 patients with AIS who initiated thoracolumbosacral orthosis treatment at Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from March, 2015 to August, 2024 and had determinable outcomes were retrospectively included. Patients were divided according to a prespecified scheme into a training set (n = 133) and a validation set (n = 58). Brace treatment failure was defined as progression of the major-curve Cobb angle by ≥ 5° from baseline during follow-up or at brace weaning, or a final major-curve Cobb angle > 45°. Logistic regression, random forest, support vector machine with radial basis function kernel, and extreme gradient boosting (XGBoost) models were developed using routinely available clinical and radiographic variables before brace treatment and during initial brace fitting. Stratified five-fold cross-validation was used for hyperparameter tuning in the training set. Model performance was evaluated in the validation set using the area under the receiver operating characteristic curve (AUC), area under the precision-recall curve (AUPRC), Brier score, calibration curves, and decision curve analysis (DCA). SHapley Additive exPlanations (SHAP) was used to interpret the main explanatory model. ResultsIn cross-validation of the training set, both random forest and XGBoost showed good discriminative performance, with cross-validation-AUC values of 0.871 and 0.880, respectively. In the validation set, the AUC values of random forest and XGBoost were 0.888 and 0.899, the AUPRC values were 0.678 and 0.830, and the Brier scores were 0.086 and 0.076, respectively. Random forest was selected as the primary model for interpretation, with a calibration intercept of 0.091 and a calibration slope of 0.995. SHAP analysis indicated that the in-brace correction rate and the major-curve Cobb angle measured in the brace contributed most to the model predictions. ConclusionThe random forest model developed using routinely available pretreatment and initial-fitting variables shows preliminary predictive value for brace treatment failure risk in AIS, and may provide quantitative support for follow-up scheduling, brace-fit reassessment and early adherence-oriented management.

BackgroundSuicide and self-injury risk in patients with depression disorder is associated with pain symptoms， but the relationships between specific pain types and their severity and distinct self-injury/suicide risk phenotypes remain controversial. ObjectiveTo explore whether somatic pain can reflect different self-injury/suicide risk phenotypes， and to provide references for the clinical identification of suicide and self-injury risk in patients with depression. MethodsBased on data from the National Survey on Symptomatology of Depression （NSSD）， 3 275 patients who met the diagnostic criteria for major depressive disorder in the Diagnostic and Statistical Manual of Mental Disorders， fourth edition， text revision （DSM-IV-TR） were recruited. According to previous suicidal behavior， suicidal ideation and self-injury behavior in the past two weeks， patients were divided into the no self-injury/suicide risk group， self-injury group， and high suicide-risk group. The occurrence of headache， muscle soreness， trunk pain， limb pain， and other pain symptoms was assessed using the structured symptom assessment instrument of the NSSD. With the no self-injury/suicide risk group as the reference， an ordinal Logistic regression model was used to analyze the associations between the risk groups and the occurrence frequency of pain， with demographic and clinical characteristics entered into the model as covariates for adjustment. ResultsA total of 3 211 patients （98.05%） were included in the analysis， including 1 329 cases （41.39%） in the no self-injury/suicide risk group， 415 cases （12.92%） in the self-injury group， and 1 467 cases （45.69%） in the high suicide-risk group. Compared with the no self-injury/suicide risk group， the occurrence frequencies of headache （OR=1.526， 95% CI： 1.221–1.894） and muscle soreness （OR=1.287， 95% CI： 1.034–1.597） were higher in the self-injury group， and the occurrence frequencies of headache （OR=0.583， 95% CI： 0.504–0.672）， muscle soreness （OR=0.682， 95% CI： 0.597–0.780）， trunk pain （OR=0.557， 95% CI： 0.475–0.634）， limb pain （OR=0.454， 95% CI： 0.384–0.522）， and other pain difficult to localize （OR=0.542， 95% CI： 0.472–0.631） were all lower in the high suicide-risk group. ConclusionPatients with depression disorder across different self-injury and suicide risk phenotypes exhibit distinct pain characteristics， and these differences may provide a reference for the clinical identification of suicide and self-injury risk in patients with depression disorder. ［Funded by The National Key Research and Development Program of China during the "13th Five-Year Plan" period （number， 2016YFC1307100）； Shanghai Key Clinical Specialties Construction Project］

ObjectiveTo investigate the effectiveness and safety of coblopasvir hydrochloride combined with sofosbuvir （CLP/SOF） in the treatment of patients with chronic hepatitis C （CHC） and those with genotype 3b in terms of virologic response rate， changes in liver and renal function parameters， and adverse events， and to provide a reference for clinical diagnosis and treatment. MethodsThe patients with CHC who were admitted to Maanshan Fourth People’s Hospital from March 1， 2023 to July 31， 2024 were enrolled and treated with CLP/SOF for a course of 12 weeks. Follow-up assessments were conducted at weeks 4， 8， and 12 of treatment and at 12 weeks after the end of treatment， and related data were collected， including HCV RNA， routine blood test results， and liver and renal function parameters. Sustained virologic response at 12 weeks after treatment （SVR12） and safety profile were evaluated. The Friedman test was used for comparison of liver and renal function parameters， routine blood test results， and liver fibrosis score at different time points. ResultsA total of 107 CHC patients treated with CLP/SOF were enrolled， with an age of 22 — 92 years， and there were 55 male patients. The predominant genotypes were 1b （46.73%） and 3b （28.97%）. Comorbidities mainly included liver cirrhosis （15 patients）， fatty liver disease （27 patients）， syphilis （15 patients）， HBV infection （8 patients）， and HIV infection （2 patients）. The overall SVR12 rate was 100% （107/107）， with an SVR12 rate of 100% for subgroups with different genotypes and those with liver cirrhosis， fatty liver disease， syphilis， HBV infection， or HIV infection. The patients with HCV genotype 3b comorbid with liver cirrhosis， fatty liver disease， syphilis， HBV infection， hypertension or diabetes also achieved an SVR12 rate of 100%. After CLP/SOF treatment， the patients showed significant reductions in the levels of the liver function parameters alanine aminotransferase， aspartate aminotransferase， total bilirubin， and hemoglobin （all P<0.001） and significant reductions in the liver fibrosis scores fibrosis-4 and aspartate aminotransferase-to-platelet ratio index （both P<0.001）. The median creatinine level was 66 μmol/L before treatment， which decreased to 52 μmol/L at week 12 of treatment （P<0.001）. No adverse events were reported. ConclusionPatients with CHC achieve a high SVR12 rate and significant improvements in liver function parameters and liver fibrosis markers after treatment with the CLP/SOF regimen， and this regimen has no significant impact on renal function and shows a favorable safety profile. Therefore， it holds promise for clinical application.

OBJECTIVE To evaluate the quality of Zhachong-13 pills.METHODS The contents of agarotetrol， costunolide， hypaconitine， dehydrocostus lactone， mesaconitine， aconitine， liquiritigenin， glycyrrhizic acid， ellagic acid， liquiritin， corilagin， and gallic acid in 18 batches of samples were determined by high-performance liquid chromatography-tandem triple quadrupole mass spectrometry （HPLC-MS/MS） method. Cluster analysis （CA） was performed using the Microbioinformatics online analysis platform. Principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were performed using SIMCA 14.1 software. Random forest， extremely randomized trees， and extreme gradient boosting machine learning models were constructed using the SPSSAU online analysis platform. The comprehensive quality of the samples was evaluated by the entropy weight-technique for order preference by similarity to ideal solution （TOPSIS） method.RESULTS The contents of the above 12 components were 104.331-694.382， 652.217-1 085.075， 6.051-130.980， 612.174-1 417.179， 1.018-32.239， 0.953-24.967， 65.552-652.239， 2 019.412-5 385.807， 52.867-4 990.450， 156.307-883.180， 10.142-475.722， and 1 102.863-4 826.396 μg/g， respectively. The results of CA and PCA showed that the 18 batches of samples were divided into two categories： S1-S6， S10-S15 were grouped into one category， while S7-S9 and S16-S18 were grouped into another category. The OPLS-DA results showed that the variable importance in projection values of mesaconitine， glycyrrhizic acid， liquiritin， ellagic acid， hypaconitine， liquiritigenin， agarotetrol， gallic acid， and dehydrocostus lactone were greater than 1. The results of the three machine learning models showed that ellagic acid and hypaconitine had relatively large differential contributions. The entropy weight-TOPSIS results showed that the overall quality of samples from manufacturer B was superior to that of the other four manufacturers.CONCLUSIONS The established method is stable and feasible， and can be used to evaluate the quality of Zhachong-13 pills， in combination with multivariate statistical analysis. Ellagic acid and hypaconitine are differential markers that affect its quality.

The recombinant human thyroid-stimulating hormone (rhTSH) can be used for differentiated thyroid cancer (DTC) preparation before ¹³¹I therapy, and follow-up. Since its approval by the American Food and Drug Administration (FDA) in 1998, rhTSH has been clinically used in European countries, the United States, and other countries for nearly 30 years. In April 2024, China’s first independently developed rhTSH agent was approved. Based on clinical experience and evidence from domestic and international studies, an expert consensus on the application of rhTSH has been developed following thorough consultations among experts. The consensus offers evidence-based recommendations to guide the appropriate use of rhTSH in clinical practice.

Background and aims:Primary sclerosing cholangitis(PSC)is an autoimmune liver disease characterized by complex pathogenesis and limited available therapeutic options.The mechanisms underlying the development and progression of PSCs remain unclear.Liver X receptor beta(LXR-β)is recognized to modulate lipid metabolism and immune response,but its specific involvement in the PSC has not been elucidated.Here,we explored the role and mechanism of LXR-β in PSC induced by 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine(DDC).Methods:CRISPR-Cas9 technology was applied to generate Abcb4(coding MDR2,next named as Mdr2),Nr1h2(coding LXR-β,next named as Lxrβ),and Rag2(coding RAG2)knockout mice.DDC was used to induce PSC.Hematoxylin and eosin and Sirius red staining were used to assess the extent of hepatic injury and fibrosis.Flow cytometry was used to observe immune cell subsets.Results:We observed a declining trend in hepatic Lxrβ in the PSC model.Unexpectedly,Lxrβ knockout failed to modulate DDC-induced PSC pathogenesis.Concomitantly,assessment of the influence of Rag2 deficiency on PSC progression revealed the absence of aggravated or alleviated hepatic injury or fibrosis in the Rag2-/-DDC mice.However,Lxrβ depletion intensified DDC-induced PSC in the Rag2-/-mice,with more abundant infiltrative inflammatory cells and more severe liver fibrosis.Compared with Rag2-/-DDC mice,Lxrβ-/-Rag2-/-DDC mice had higher serum ALT and AST levels and mRNA expression of proinflammatory and profibrotic genes.Flow cytometry showed that LXR-β deficiency resulted in a diminished population of hepatic innate immune cells.Conclusion:This study indicated innate immune cell LXR-β deficiency can exacerbate hepatic injury and fibrosis in murine models of PSC suggesting that LXR-β may regulate the function of innate immunity in the fibrotic advancement of PSC.

Objective To determine the median effective dose(ED50)of remimazolam for preoperative sedation in pediatric patients aged 1-6 years using the modified Dixon sequential method.Methods This is a prospective clinical study.Pediatric patients scheduled for elective short surgery(surgery time≤1 h)under general anesthesia from January to July 2023 were selected.Inclusion criteria were age 1-6 years,an ASA physical status Ⅰ-Ⅱ and the preoperative parent separation anxiety scale(PSAS)score≥3 points.Remimazolam was administered intravenously preoperatively,and its sedative effect was assessed.The modified Dixon sequential method was used to determine the ED50 of remimazolam,with the initial dose set at 0.10 mg/kg and the dose increment set at 0.02 mg/kg.Sedation was considered successful(positive,included in positive group)if the child with sedation score≥2 points,preoperative PSAS score<3 points,and the mask acceptance score of 4 points during anesthesia induction.If any criterion was not met,sedation was considered failure(negative,included in negative group),and the next patient's dosage was increased by 0.02 mg/kg based on the previous patient's dosage.The test was completed after 7 consecutive positive and negative turning points appeared alternately.Probabilistic unit regression analysis was used to determine the ED50,ED95 and the corresponding 95%confidence interval(CI)of remimazolam for preoperative sedation.Postoperative recovery time and adverse events such as airway spasm,respiratory depression,hypotension,nausea and vomiting during anesthesia were recorded.Results A total of 23 pediatric patients were included,with 13 in positive group and 10 in negative group.There were no statistically significant differences in mean arterial pressure,pulse oxygen saturation or heart rate before and after sedation(P>0.05).Compared with negative group,positive group showed a significant reduction in preoperative parent separation anxiety and an increase in mask acceptance during anesthesia(P<0.05).There was no significant difference in sedation score and anesthesia awakening time between two groups(P>0.05).The ED50 of remimazolam for preoperative sedation in pediatric patients aged 1-6 years was 0.051 mg/kg(95%CI 0.033-0.065 mg/kg),and the ED95 was 0.077 mg/kg(95%CI 0.064-0.161 mg/kg).No adverse events such as airway spasm,respiratory depression,hypotension,nausea and vomiting occurred during anesthesia in any of pediatric patients.Conclusion The ED50 of intravenous administration of remimazolam for preoperative sedation in pediatric patients aged 1-6 years is 0.051 mg/kg(95%CI 0.033-0.065 mg/kg).