An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective To test the hypothesis that backward crawling and forward crawling share similar inter-joint coordination patterns,thus providing potential evidence for the application of backward crawling in rehabilitation training.Methods The acceleration signals in the X,Y,and Z directions for 9 joints(including bilateral wrists,elbows,shoulders,knees,and hips)in 9 volunteers during forward and backward crawling were collected using a custom signal acquisition system,and the pressure signals were also recorded when the palms contacted the ground.The collected acceleration signals were preprocessed,segmented into cycles,and vectorized.Based on the pressure signals,a single crawling cycle was divided into support phase and swing phase.In addition,principal component analysis was applied to extract inter-joint coordination in limbs at various scales(sagittal,coronal,and transverse planes).Pearson correlation coefficients of inter-joint coordination patterns were compared between forward and backward crawling in support period,swing period,and full cycle.Results The correlation coefficients for coordination patterns in the full cycle at the transverse plane scale were 0.813 5(PC1)and 0.837 5(PC2),and the correlation coefficient of the support period PC2 was 0.901 8.At the sagittal plane scale,the correlation coefficient of the support period PC1 was 0.948 5.Conclusion The study provides preliminary evidence that limb motion coordination patterns during backward crawling are similar to those observed during forward crawling.Future research will further explore the effects of backward crawling on functional rehabilitation in individuals with motor impairments.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

Objective To survey the free influenza vaccination status and willingness among the elderly in Deqing County,Zhejiang Province,and analyze the influencing factors.Methods Based on population data and the electronic medical system of Deqing County,the influenza vaccine coverage from 2018 to 2022 after the implementation of the free influenza vaccination policy was calculated.A proportional stratified random sampling method was used to select elderly individuals aged 60 and above from six streets/towns in Deqing County.A questionnaire survey was conducted to assess influenza vaccination behaviors from 2020 to 2022 and vaccination willingness for the 2023 influenza season.The chi-square test and logistic regression were used to analyze potential factors associated with vaccination behavior and subsequent willingness among the elderly.Results An average of influenza vaccines for the elderly per year were 56.2 thousand,with influenza vaccination coverage rate of 46.16%in average over the five years.Among 729 subjects,the vaccination rate increased from 69.3%in 2020 to 75.3%in 2022,and 81.9%of them were willing to receive influenza vaccines in the next flu season.Logistic regression analysis revealed that individuals with agricultural household registration(OR=3.05,95%CI:1.82-8.99),lower education levels(OR=2.82,95%CI:1.15-6.93),awareness of the free vaccine policy(OR=4.65,95%CI:1.82-11.91),and belief in vaccine efficacy(OR=6.61,95%CI:3.20-13.61)had higher vaccination rates.Factors associated with a more positive vaccination willingness included an annual household income of<150 thousand CNY(OR=1.65,95%CI:1.04-2.60),awareness of the free vaccine policy(OR=3.07,95%CI:1.84-5.12),perceived vaccine efficacy(OR=4.39,95%CI:2.77-6.96),and perceived vaccine safety(OR=1.57,95%CI:1.01-2.43).The main reasons for non-vaccination were concerned about the side effects of the vaccine(37.9%),thoughts that they were in good health and did not need to be vaccinated(30.3%),and influenza vaccines were ineffective(21.2%).The main reasons for vaccination were to protect themselves(89.1%),vaccination was free(65.5%)and to protect their families(60.6%).Conclusion The elderly in Deqing County had a relatively high influenza vaccination rate and follow-up vaccination willingness rate,but there still existed a gap to reach at target rate recommended by WHO.The local government should strengthen health education related to influenza vaccine knowledge,and publicize free vaccination policy to improve the vaccination rate and willingness in next season.

Objective:To develop and validate machine learning models for predicting the cumulative live birth rate (CLBR) following in vitro fertilization (IVF) and to analyze key predictive features using SHAP values. Methods:This retrospective study included data from patients who underwent IVF-embryo transfer at the Department of Reproductive Medicine, Baoding Maternal and Child Health Hospital, between January 2017 and December 2022. Patients were categorized into two groups based on live birth outcome: the live birth group ( n=1 036) and the non-live birth group ( n=756). The dataset was randomly divided into a training set and a validation set in a ratio of 7∶3. Five algorithms were utilized for model development: logistic regression, random forest, extreme gradient boosting (XGBoost), support vector machine, and neural networks. Model performance was assessed using the area under the receiver operating characteristic (AUC) curve, F1 score, and calibration curves. Clinical decision curve analysis (DCA) was employed to evaluate the clinical utility of the models. SHAP values were used to interpret feature importance in the XGBoost model and enhance its explainability. Results:The XGBoost model demonstrated the best performance in predicting CLBR,with accuracy of 72.44%, AUC of 0.775, and F1 score of 0.654, accuracy and F1 score outperforming logistic regression (accuracy was 70.02%, F1 score was 0.585), random forest (accuracy was 71.69%, F1 score was 0.606), support vector machine (accuracy was 70.20%, F1 score was 0.607), and neural network (accuracy was 68.72%, F1 score was 0.560). The calibration curve of XGBoost closely aligned with the diagonal line, indicating that the predicted probabilities were very close to the actual outcomes, demonstrating good calibration. DCA indicated that the XGBoost model provided higher net benefits across a wide range of clinical decision thresholds. SHAP value analysis identified number of previous IVF failures, antral follicle count, anti-Müllerian hormone level, percentage of normal sperm morphology, and sperm DNA fragmentation index as key predictors of CLBR.Conclusion:The XGBoost model exhibits excellent predictive performance and calibration for CLBR, with SHAP values providing important insights into feature importance. This model has the potential to support the development of personalized treatment strategies in clinical practice. However, its generalizability needs to be validated using external datasets to ensure its applicability to diverse populations.

Objective:To investigate the clinical efficacy of compound sabal berry tablets on overactive bladder symptoms in patients with benign prostatic hyperplasia after transurethral laser enucleation of the prostate.Methods:This study was a prospective study. Eighty patients with benign prostatic hyperplasia who underwent laser enucleation at Tongliao People's Hospital from January 2024 to December 2024 were included in this study. The patients were randomly divided into a study group and a control group using the random number table method, with 40 patients per group. The control group received 0.2 mg of tolterodine tartrate tablets twice a day after surgery. The study group was given compound sabal berry tablets (0.5 g orally three times a day) in addition to the treatment provided to the control group. Both groups of patients were treated for 4 weeks after surgery. The clinical efficacy of the two groups was compared, including the International Prostate Symptom Score (IPSS), Overactive Bladder Symptom Score (OABSS), Maximum Postoperative Urinary Flow Rate (Qmax), Post-Void Residual (PVR), and the incidence of postoperative bladder irritative symptoms.Results:The differences in the preoperative indicators, including IPSS, OABSS, Qmax, and PVR, between the two groups were not statistically significant (all P > 0.05). Preoperatively, in the control group, Qmax was (8.64 ± 2.83) mL/s, IPSS was (25.10 ± 4.37), OABSS was (10.52 ± 1.87), and PVR was (80.70 ± 6.34) mL; in the study group, the respective values were (9.12 ± 2.95) mL/s, (24.60 ± 4.53), (10.83 ± 1.73), and (80.10 ± 5.61) mL. Postoperatively, in the control group, Qmax was (20.30 ± 3.65) mL/s, IPSS was (8.50 ± 1.58), OABSS was (4.09 ± 0.52), and PVR was (9.70 ± 2.48) mL, while in the study group, the respective values were (21.40 ± 4.38) mL/s, (7.40 ± 1.76), (1.71 ± 0.36), and (9.00 ± 1.75) mL. Postoperatively, both groups showed a significant increase in Qmax, while IPSS, OABSS, and PVR all significantly decreased (all P < 0.05). Postoperatively, the IPSS and OABSS in the study group were significantly lower than those in the control group ( t = -3.28, -25.89, both P < 0.05). However, there were no statistically significant differences in Qmax and PVR between the two groups (both P > 0.05). The incidence of bladder irritative symptoms in the study group [12.50% (5/40)] was significantly lower than that in the control group [35.00% (14/40), χ2 = 8.64, P < 0.05]. Conclusions:Compound sabal berry tablets can reduce postoperative prostate symptoms and overactive bladder symptoms in patients undergoing transurethral laser enucleation of the prostate for benign prostatic hyperplasia, demonstrating a certain clinical efficacy.

Objective To understand the epidemiological characteristics and trends of postoperative pneumonia(POP)in tertiary general hospitals in Jiangsu Province,and provide theoretical basis for carrying out targeted pre-vention and control measures.Methods Surgery patients from 22 tertiary general hospitals in 12 cities in north,central,and south of Jiangsu Province from January 1,2022 to December 31,2023 were chosen as studied subjects,occurrence of POP was analyzed and compared.Results A total of 848 274 surgical procedures were performed in 22 hospitals,and 3 606 cases of POP occurred,with an incidence of 0.43％.The incidence in 2023 was 0.37％,which was lower than that in 2022(0.49％),with statistically significant difference(P＜0.001).The top three de-partments with high incidence of POP were neurosurgery(6.71％),cardiothoracic surgery(2.91％),and general surgery(0.77％).Among hospitals of different grades,the incidence of POP in tertiary first-class hospitals was 0.44％,which was higher than that in other tertiary hospitals(0.37％).There was no statistically significant difference in the incidence of POP between municipal and district/county hospitals(P＞0.05).The incidence of POP in hospitals with a bed:infection control full-time staff ratio＜200∶1 was lower than that in hospitals with the ratio ≥200∶1(0.39％vs 0.47％,P＜0.001),while the incidence of POP in hospitals with a proportion ≥30％of full-time staff being doctors was higher than that in hospitals with a proportion＜30％(0.45％vs 0.36％,P＜0.001).The incidence of POP in male patients was higher than that in female patients(0.62％vs 0.26％,P＜0.001).The incidence of POP in elderly patients aged≥65 was higher than that in patients aged＜65(0.73％vs 0.26％,P＜0.001).A total of 2 667 strains of infectious pathogens were detected,with the top three being Acine-tobacter baumannii,Klebsiella pneumoniae,and Pseudomonas aeruginosa,accounting for 28.95％,22.72％,and 15.45％,respectively.The detection rates of carbapenem-resistant Acinetobacter baumannii(CRAB),carba-penem-resistant Klebsiella pneumoniae(CRKP),and carbapenem-resistant Pseudomonas aeruginosa(CRPA)were 60.75％,21.45％,and 32.28％,respectively.The detection rate of CRKP decreased in 2023 compared with 2022,with statistically significant difference(P＜0.05).Conclusion The overall incidence of POP in tertiary general hos-pitals in Jiangsu Province is relatively low,but there are significant differences among different hospitals.There-fore,perioperative prevention and control measures should be carried out based on the epidemiological characteristics of patients.

To promote the effective implementation of China's Essential Public Health Services Program(EPHSP)and ensure the secure and efficient utilization of project funds,China officially initiated the performance evaluation of EPHSP in 2011.This performance evaluation has evolved through three distinct phases:initial exploration(2011-2014),steady advancement(2015-2018),and reform and enhancement(2019—present).The evaluation objectives have progressively expanded from an initial focus on fund security and service coverage to a broader emphasis on enhancing service quality,improving residents'health status and sense of benefit,and facilitating the refinement of policy frameworks and the implementation of primary responsibilities.Performance evaluation has emerged as a critical instrument for strengthening project governance and optimizing resource allocation,gradually establishing a performance-driven incentive mechanism that aligns rewards with both the quantity and quality of work.This approach has effectively contributed to the continuous improvement of service quality.To further advance the high-quality development of EPHSP,future efforts should optimize the performance evaluation system,prioritize the adoption of a full-cycle performance management approach and the integration of health outcome-based indicators.Additionally,it is essential to deepen the application of information technologies to enhance the precision and efficiency of evaluations,and to innovate mechanisms for utilizing evaluation results to reinforce accountability at the local level.These measures will collectively strengthen project performance management capabilities.

Purpose: Cancer has become a significant major public health concern, making the discovery of new cancer markers or therapeutic targets exceptionally important. Elevated expression of tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) expression has been observed in certain types of cancer. This project aims to investigate the function of TNFRSF12A in tumors and the underlying mechanisms. Materials and Methods: Various websites were utilized for conducting the bioinformatics analysis. Tumor cell lines with stable knockdown or overexpression of TNFRSF12A were established for cell phenotyping experiments and subcutaneous tumorigenesis in BALB/c mice. RNA-seq was employed to investigate the mechanism of TNFRSF12A. Results: TNFRSF12A was upregulated in the majority of cancers and associated with a poor prognosis. Knockdown TNFRSF12A hindered the colorectal cancer progression, while overexpression facilitated malignancy both in vitro and in vivo. TNFRSF12A overexpression led to increased nuclear factor кB (NF-κB) signaling and significant upregulation of baculoviral IAP repeat containing 3 (BIRC3), a transcription target of the NF-κB member RELA, and it was experimentally confirmed to be a critical downstream factor of TNFRSF12A. Therefore, we speculated the existence of a TNFRSF12A/RELA/BIRC3 regulatory axis in colorectal cancer. Conclusion TNFRSF12A is upregulated in various cancer types and associated with a poor prognosis. In colorectal cancer, elevated TNFRSF12A expression promotes tumor growth, potentially through the TNFRSF12A/RELA/BIRC3 regulatory axis.

Learning-associated functional plasticity at hippocampal synapses remains largely unexplored. Here, in a single session of reward-based trace conditioning, we examine learning-induced synaptic plasticity in the dorsal CA1 hippocampus (dCA1). Local field-potential recording combined with selective optogenetic inhibition first revealed an increase of dCA1 synaptic responses to the conditioned stimulus (CS) induced during conditioning at both Schaffer collaterals to the stratum radiatum (Rad) and temporoammonic input to the lacunosum moleculare (LMol). At these dCA1 inputs, synaptic potentiation of CS-responding excitatory synapses was further demonstrated by locally blocking NMDA receptors during conditioning and whole-cell recording sensory-evoked synaptic responses in dCA1 neurons from naive animals. An overall similar time course of the induction of synaptic potentiation was found in the Rad and LMol by multiple-site recording; this emerged later and saturated earlier than conditioned behavioral responses. Our experiments demonstrate a cued memory-associated dCA1 synaptic plasticity induced at both Schaffer collaterals and temporoammonic pathways.
Animals ; CA1 Region, Hippocampal/physiology* ; Male ; Association Learning/physiology* ; Neuronal Plasticity/physiology* ; Cues ; Memory/physiology* ; Synapses/physiology* ; Conditioning, Classical/physiology* ; Excitatory Postsynaptic Potentials/physiology* ; Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors* ; Rats ; Optogenetics