Objective: To explore the association between the HLA antibody positivity rate in female blood donors and pregnancy history, number of pregnancies, interval from the last pregnancy to blood donation, and age, to identify associated variables using a univariate generalized additive model (GAM), and to further analyze the predictive role of characteristic variables for HLA antibody positivity using a random forest model. Methods: HLA antibody detection was performed on 391 female blood donors using the Luminex immunomagnetic bead method. The correlation between pregnancy-related factors and HLA antibodies was analyzed using the Chi-square test. Based on R software, a univariate GAM was first constructed to analyze the association types between characteristic variables and the HLA antibody positivity rate, followed by the construction of a random forest model to evaluate the predictive value of the variables. Results: Among the 391 female blood donors without a transfusion history, the overall HLA antibody positivity rate was 26.34%. The positivity rate in donors with a pregnancy history was significantly higher than that in those without (30.09% vs 9.72%, P<0.05), and HLA antibody positivity rate increased linearly with the number of pregnancies (P<0.05). In the univariate GAM, age and number of deliveries exhibited a non-linear association with the HLA antibody positivity rate (the positivity rate increased sharply between 25-35 years of age and stabilized after 3 deliveries). Besides, the interval from the last pregnancy to blood donation showed a linear association with the HLA antibody positivity rate, and the positivity rate decreased as the interval prolonged (P<0.05). In the random forest model, age (mean decrease gini=29.26) and interval from the last pregnancy to blood donation (mean decrease gini=22.02) were core predictive variables: age was more conducive to identifying positive samples, while the interval from the last pregnancy to blood donation was more helpful for excluding negative samples. The number of deliveries (mean decrease accuracy=16.98) made a significant contribution to predicting positive samples, whereas the number of abortions had no impact. The model had an AUC of 0.583 (95% CI: 0.593 8-0.770 2), indicating a certain predictive value. Conclusion: The associated variables identified by the univariate GAM model, including age, interval from the last pregnancy to blood donation, and number of deliveries, provide a basis for key variables in the random forest model. All three variables have predictive value for HLA antibody positivity, which can provide evidence-based support for personalized transfusion management and stratified screening of female blood donors in this region.

ObjectiveTo investigate the clinical effects and safety of Shuhe Granules （舒和颗粒） in patients with rheumatoid arthritis （RA） comorbid with insomnia. MethodsA single-arm prospective exploratory design was adopted. Thirty RA patients comorbid with insomnia and diagnosed with the traditional Chinese medicine （TCM） syndrome of kidney-heart deficiency and qi-blood disharmony were enrolled. On the basis of maintaining their original RA treatment regimens， all patients received oral Shuhe Granules， one dose per day， for 12 consecutive weeks. The outcomes were assessed before treatment and at weeks 4， 8， and 12 after treatment. Sleep and related psychosomatic function were evaluated using the insomnia severity index （ISI）， Pittsburgh sleep quality index （PSQI）， fatigue severity scale （FSS）， patient health questionnaire-9 （PHQ-9）， generalized anxiety disorder-7 （GAD-7）， and Montreal cognitive assessment （MoCA） scores. RA-related assessments included the 28-joint disease activity score based on C-reactive protein （DAS28-CRP）， pain visual analog scale （VAS）， tender joint count （TJC28）， swollen joint count （SJC28）， morning stiffness duration， and health assessment questionnaire （HAQ） score. Laboratory parameters included erythrocyte sedimentation rate （ESR）， serum C-reactive protein （CRP）， anti-cyclic citrullinated peptide antibody （anti-CCP）， and rheumatoid factor （RF） levels. A literature-based historical control group was established through literature retrieval and meta-analysis. The ISI， PSQI， DAS28-CRP， and pain VAS scores were compared between the treatment group in the present study and the historical control group. Safety assessments were also performed. ResultsCompared with baseline before treatment， ISI and PSQI scores decreased at weeks 4， 8， and 12， while MoCA scores increased at weeks 4， 8， and 12. FSS scores decreased at weeks 4 and 12. At weeks 8 and 12， GAD-7 scores， DAS28-CRP scores， duration of morning stiffness， TJC28， and SJC28 were reduced. At week 12， PHQ-9， pain VAS， and HAQ scores significantly decreased （P＜0.05）. FSS and DAS28-CRP scores at week 12 were lower than those at week 4， and CRP levels at week 8 were lower than those at week 4 （P＜0.05）. Compared with the historical control group， the treatment group showed superior improvement in ISI and PSQI scores （P＜0.001）； however， no statistically significant differences were observed between the two groups in DAS28-CRP or pain VAS scores （P＞0.05）. No serious adverse events occurred during the study， and the 4 reported mild adverse events were all considered unrelated to the study medication. ConclusionShuhe Granules significantly improve sleep quality， fatigue， emotional status， cognitive function， RA-related symptoms， and joint function in patients with RA comorbid insomnia， with good safety and tolerability.

The Split-Cre system consists of two inactive polypeptides: NCre and CCre, which can be recombined into an active full-length Cre under certain conditions. This system is typically used with LoxP. To develop an efficient Split-Cre system, this study used Rma intein from Rhodothermus marinus to split Cre and screened out the split site S102 which could efficiently mediate the recombination of Cre in the "Traffic Light" reporter cell line. Moreover, the S102 Split-Cre system was delivered to mice by dual-adeno-associated virus (AAV), and it was demonstrated that the efficiency of the Rma intein-mediated S102 Split-Cre system was comparable to the full-length Cre in mice. This system lays a foundation for both basic and applied research on Split-Cre.
Inteins/genetics* ; Animals ; Integrases/biosynthesis* ; Mice ; Dependovirus/metabolism* ; Bacterial Proteins/genetics* ; Recombination, Genetic ; Humans

ObjectiveTo analyze the evolutionary characteristics and genetic variations of the HA (hemagglutinin) and NA (neuraminidase) genes of influenza A(H1N1) viruses in Shanghai during 2024, to investigate their transmission patterns, and to evaluate their potential impact on vaccine effectiveness. MethodsFrom January to October 2024, throat swab specimens were collected from influenza like illness (ILI) patients at 4 hospitals in Shanghai. Real-time fluorescence ploymerase chain reaction (RT-PCR) was used for virus detection and isolation of H1N1 influenza viruses. Forty influenza A(H1N1) virus strains were sequenced using Illumina NovaSeq 6000 platform, followed by phylogenetic analyses, genetic distance analysis, and amino acid variation analyses of HA and NA genes. ResultsPhylogenetic tree of the HA and NA genes revealed that the 40 influenza A(H1N1) virus strains circulating in Shanghai in 2024 exhibited no significant geographic clustering, with a broad origin of strains and complex transmission chains. Genetic distance analyses demonstrated that the average intra-group genetic distances of HA and NA genes among the Shanghai strains were 0.005 1±0.000 6 and 0.004 6±0.000 6, respectively, which were comparable to or higher than those observed in global surveillance strains. Both HA and NA genes displayed frequent mutations. Compared to the 2023‒2024 and 2024‒2025 Northern Hemisphere A(H1N1) vaccine strains (WHO-recommended), the HA proteins of 40 Shanghai strains exhibited amino acid substitutions at positions 120, 137, 142, 169, 216, 223, 260, 277, 356 and 451, with critical mutations at positions 137 and 142 located within the Ca2 antigenic determinant. Furthermore, mutations in the NA protein were observed at positions 13, 50, 200, 257, 264, 339 and 382. ConclusionThe genetic background of the 2024 Shanghai influenza A(H1N1) virus strains is complex and diverse, and antigenic variation may affect vaccine effectiveness. Therefore, it is recommended to enhance genomic surveillance of influenza viruses, evaluate vaccine suitability, and implement more targeted prevention and control strategies against imported influenza viruses.

OBJECTIVE To observe the clinical efficacy of Zhenshi Burn Ointment on postoperative wound healing of simple anal fistula and explore its potential mechanism of action.METHODS A total of 60 patients with low-position simple anal fistula of damp-heat pouring downward type admitted to the Department of Anorectum,Nanjing Hospital of Chinese Medicine Affiliated to Nan-jing University of Chinese Medicine from May to December 2023 were selected as the research subjects and randomly divided into an experimental group and a control group,with 30 cases in each group.After surgery,the control group was given sterilized vaseline gauze for external application,and the experimental group was given Zhenshi Burn Ointment for external application.The course of treatment for both groups was 30 d.After treatment,the postoperative wound healing time,wound healing rate,pain score,wound granulation growth and secretion,and changes in TCM syndrome score were compared between the two groups.The pathological sec-tions of wound granulation tissue were compared 14 d after surgery.The transcription levels of COL1A1,type Ⅲ collagen(COL3A1),epidermal growth factor(EGF),and fibroblast growth factor(FGF2)in granulation tissue were detected by qPCR.The expression lev-els of proteins related to MEK/ERK signaling pathway were detected by Western blot.RESULTS Compared with the control group,the wound healing rates of the experimental group were significantly increased at 7,14,21,and 30 d after surgery(P<0.01),and the wound healing time was significantly shortened(P<0.01).At 7 d after surgery,the pain score of the experimental group was signifi-cantly lower than that of the control group(P<0.05).At 7 and 14 d after surgery,the granulation growth score of the wound in the ex-perimental group was significantly lower than that of the control group(P<0.01).At 14 d after surgery,the secretion score of the ex-perimental group was significantly lower than that of the control group(P<0.01).At 14 and 30 d after surgery,the total score of TCM syndrome in the experimental group was significantly better than that in the control group(P<0.05).At 14 d after surgery,the patho-logical improvement of granulation tissue in the wound in the experimental group was better than that in the control group,and the con-tent of collagen fiber and type I collagen(Collagen I)was significantly higher than that in the control group(P<0.01).The mRNA contents of COL1A1,COL3A1,EGF,and FGF2 were significantly higher than those in the control group(P<0.05,P<0.01);the levels of p-MEK1/2 and p-ERK1/2 proteins were higher than those in the control group(P<0.05).CONCLUSION Zhenshi Burn Ointment has a good effect on promoting postoperative anal fistula wound healing,which may be related to upregulating growth factor expression,promoting collagen regeneration,and activating MEK/ERK pathway.

Objective To investigate the drug-drug interaction between donepezil hydrochloride and moxifloxacin hydrochloride during combined administration through in vitro liver microsomal metabolism and changes in pharmacokinetic characteristics in rats.Methods A rat liver microsomal incubation system was constructed and optimized.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)analysis of donepezil hydrochloride and high-performance liquid chromatography(HPLC)analysis of moxifloxacin hydro-chloride were performed.The pharmacokinetic and metabolic characteristics of the two drugs,alone and in combination,were compared in vitro using rat liver microsomes and in vivo using rats.Results In the liver microsomal system,the half-life(T1/2)of donepezil hydro-chloride in the combined administration 1 group was prolonged by 29.892％(P＜0.01)and the intrinsic clearance(CLint)was reduced by 23.194％(P＜0.01)compared with the donepezil hydrochloride group.Conversely,the metabolic parameters of moxifloxacin hydrochlo-ride in the combined administration 2 group did not differ significantly from that of the moxifloxacin hydrochloride group(P＞0.05).In the in vivo study,the AUC0～t and AUC0～∞ of donepezil hydrochloride in the combined administration 1 group increased by 44.259％and 44.496％,respectively,maximum plasma concentration(Cmax)increased by 117.723％,T1/2 was prolonged by 98.063％,and CLint was reduced by 35.293％,compared with that in the donepezil hydrochloride group.Moreover,the differences were all statistically significant(P＜0.05).The in vivo study findings were consistent with the results of the in vitro study.Conclusion A drug-drug interaction occurs when moxifloxacin hydrochloride is used in combination with donepezil hydrochloride.Moxifloxacin hydrochloride promotes the absorption of donepezil hydrochloride,inhibits its metabolism,slows its CLint,and increases donepezil exposure in the body.

Objective:To construct fused cancer cell/neutrophil membrane-coated polydopamine nanoparticles chelated with manganese ions (Ⅱ) (PMNP@FMs) and explore the potential for targeted pancreatic cancer fluorescence imaging and MRI.Methods:Cancer cell membranes fused with neutrophil membranes were encapsulated on the surface of polydopamine nanoparticles chelated with manganese ions (Ⅱ) (PMNPs) to prepare PMNP@FMs. The morphology, structure, and MRI performance of the product were characterized. The cytotoxicity of PMNP@FMs towards human pancreatic cancer cells (PANC-1) and normal human pancreatic ductal epithelial cells (hTERT-HPNE) was evaluated using cell counting kit (CCK)-8, and in vivo toxicity was assessed in healthy mice. PANC-1 pancreatic cancer xenograft nude mouse models were established for in vivo fluorescence imaging and MRI. Data were analyzed using the independent-sample t test, repeated measures analysis of variance and the least significance difference method. Results:PMNP@FMs exhibited a core-shell structure with a diameter of (112.81±8.64) nm, negative surface charge, and good dispersibility. The T 1 relaxivity of PMNPs was 18.81±0.22, which was 4.1 times higher than that of gadopentetate dimeglumine (Gd-DTPA) (4.55±0.24; t=75.54, P<0.001). Co-culture of PMNPs and PMNP@FMs with hTERT-HPNE and PANC-1 cells for 24 h resulted in cell viability above 90% within the concentration range of 0-500 μg/ml. PMNP@FMs did not affect mouse survival and showed no apparent organ damage. In vivo fluorescence imaging and MRI revealed that PMNP@FMs accumulated highly in tumors and reached the peak 24 h post intravenous administration (relative MR signal: 1.35±0.01, fluorescence intensity: (1.20±0.25)×10 10), surpassing the peak observed in the control group (1.22±0.01, (3.87±0.50)×10 9;F values: 11.03-188.01, t values: 18.20, 5.64, all P<0.05), with hepatic metabolism being the primary route of clearance. Conclusion:PMNP@FMs demonstrate a potential for targeted pancreatic cancer fluorescence imaging and MRI, offering promising prospect for precise diagnosis of early-stage pancreatic cancer.

Objective To explore the factors influencing cognitive frailty in elderly inpatients with chronic heart failure(CHF)during hospitalisation,8 prediction models were developed with various machine learning algorithms to identify the best model as a guidance for medical staff on clinical interventions.Methods Convenience sampling method was used to select 650 elderly CHF inpatients who stayed in our hospital between September 2023 and June 2024 as the study objects in the cross-sectional investigation.A total of 607 patients had completed the study.The patients were divided into a cognitive frailty group and a non-cognitive frailty group according to the presence or absence of cognitive frailty.Variables were initially screened using univariate analysis and stepwise Logistic regression.The total sample was then randomly divided into a training set(n=424)and a testing set(n=183)of a 7:3 ratio.Eight predictive models were created using the algorithms of neural network(NN),k-nearest neighbour(KNN),linear discriminant analysis(LDA),support vector machine(SVM),naive Bayes(NB),logistic regression,decision tree(DT)and random forest(RF)on the training set.The predictive performance of the models was compared using the data of the testing set.Results The prevalence of cognitive frailty in elderly CHF inpatients was 48.3%.Results of Logistic regression showed that age,marital status,education,body mass index,multi-morbidity,nutritional status,medication,frequency of weekly exercise and the living conditions were the key factors(P<0.05).The overall accuracy in classification of the eight predictive models ranged from 0.803 to 0.847,with F1-values of 0.778 to 0.833,precision of 0.848 to 0.897,and recall rate of 0.700 to 0.778.The area under the receiver operating characteristic curve was 0.820 to 0.901.Conclusion Of the eight predictive models,the prediction model created with LDA shows the best performance and prediction in terms of comprehensive prediction metrics,while the prediction model created with NN shows the worst performance in comprehensive prediction.

Objective To explore and analyze the adverse drug event(ADE)signals associated with pembrolizumab and to provide a reference for the real-world safety of drug use on elderly patients.Methods ADE reports of pembrolizumab in the general population and elderly population were collected from the FDA Adverse Event Reporting System(FAERS)for the period between January 1st,2019,and June 30th,2024.Multiple signal detection methods(ROR,PRR,MHRA)were employed for data mining.Classification and statistical analysis were performed using the System Organ Class(SOC)and Preferred Term(PT)from the MedDRA(Version 27.1)dictionary.Results A total of 966 signals were identified in the general population,spanning 24 SOCs,while 593 signals were detected in the elderly population,spanning 23 SOCs.Comparative visualization analysis of critical PTs under four major SOCs revealed no significant abnormal signals in elderly patients.Conclusion A comprehensive and multidimensional analysis of the ADE data from the FAERS database indicates that pembrolizumab appears to be relatively safe for use in elderly patients.

In order to fully consider the physiological characteristics and movement mechanism of the human body under the premise of ensuring the function and practicality of the product,the human-fabric contact finite element model based on biomechanical feedback plays an important role in the design of'people-oriented'health protection products.This review focuses on the design of protective products made of flexible materials,and discusses the application status and development trend of human finite element model in the design of protective products.The construction process of finite element models of different parts of the human body in recent years is summarized from the perspective of human biomechanics.Secondly,from the contact models established between the human head,torso,upper limbs and lower limbs and protective devices,the application status and challenges of finite element method in the design of health protective products are sorted out.Finally,the problems existing in the use of finite element method in such researches are discussed.It is pointed out that in the context of pursuing accuracy,real-time and realism,finite element contact models with the advantages of high efficiency,precision and reusability still have a broad application prospect.