OBJECTIVE To investigate the influence of CYP2C19 gene polymorphism on platelet function and inflammatory cytokines in elderly patients with ischemic stroke， and to analyze potential factors associated with poor prognosis. METHODS A retrospective study was conducted on elderly patients with ischemic stroke admitted to our hospital from June 2024 to June 2025， wh o underwent CYP2C19 genotype testing and received antiplatelet therapy with clopidogrel. The levels of platelet function indicators and inflammatory cytokines before and after treatment were compared among patients with different metabolic phenotypes. Based on the prognosis at 6 months post-treatment， patients were divided into poor prognosis group and good prognosis group. Univariate analysis was performed on general data， metabolic phenotype， the levels of platelet function indicators and inflammatory cytokines. Variables with P ＜0.05 and the levels of inflammatory cytokines before treatment were included in a multivariate Logistic regression analysis to identify independent risk factors for poor prognosis. Multiple linear regression was used to further analyze the relationship between metabolic phenotypes and inflammatory cytokines. RESULTS A total of 448 elderly patients with ischemic stroke were included； among them， 162 cases were normal metabolic phenotype， 218 were intermediate metabolic phenotype， and 68 were poor metabolic phenotype. No rapid or ultrarapid metabolic phenotypes were observed. After treatment， platelet aggregation rate， the levels of P-selectin and platelet activated complex-1 （PAC-1）， high-sensitivity C-reactive Protein （hs-CRP）， interleukin-1β （IL-1β）， IL-6 and tumor necrosis factor-α （TNF-α） in the normal metabolic phenotype group， intermediate metabolic phenotype group， and poor metabolic phenotype group （except for platelet aggregation rate， and the levels of P-selectin and PAC-1 in the poor metabolic phenotype group） were significantly lower than those before treatment in the same group. Moreover， the above indicators in the normal metabolic phenotype group were significantly lower than those in the intermediate and poor metabolic phenotype groups at the corresponding time， and the levels of platelet function indicators in the intermediate metabolic phenotype group were significantly lower than those in the poor metabol ic phenotype group at the corresponding time （ P ＜0.05）. Univariate and multivariate Logistic regression analyses showed that combined with hypertension， combined with diabetes mellitus， and intermediate or poor metabolic genotypes were independent risk factors for poor prognosis in elderly patients with ischemic stroke （ P ＜0.05）. Multiple linear regression analysis showed that serum levels of hs-CRP， IL-1β， IL-6 and TNF-α before treatment were significantly higher in patients with intermediate and poor metabolic genotypes compared to those with normal metabolic genotype （ P ＜0.05）， with a greater magnitude of increase in inflammatory cytokines observed in the patients with poor metabolic genotype. CONCLUSIONS The elderly ischemic stroke patients with CYP2C19 intermediate and poor metabolic genotypes have poor inhibition effect on platelet and higher levels of inflammatory cytokines than normal metabolic genotype； CYP2C19 gene polymorphism， and in combination with hypertension and diabetes， can be used as independent predictors of poor prognosis.

ObjectiveIdentification and analysis, animal regression test, and drug susceptibility study were conducted on a pathogenic strain causing diarrhea in cynomolgus monkeys, aiming to provide a practical basis for the clinical treatment of Citrobacter freundii. MethodsFresh fecal samples were collected from nine diarrheal cynomolgus monkeys and streaked on Salmonella-Shigella (SS) medium, LB medium, and Columbia blood agar medium, and incubated at 37 ℃ for 24 h. Subsequently, the isolated strain was identified by colony morphology observation, Gram staining, biochemical tests and 16S rRNA gene sequencing, and PCR was used to detect its major virulence genes. After propagation, the isolate was intragastrically administered to healthy cynomolgus monkeys and C57BL/6 mice for animal regression tests to evaluate its pathogenicity. Finally, the disk-diffusion method was used to detect the antimicrobial susceptibility of the isolate. ResultsA single bacterial strain, designated MF071743, was isolated from 9 fecal samples collected from diarrheal cynomolgus monkeys. The isolate formed pink, smooth, round colonies on SS medium; translucent, smooth, moist, shiny colonies with neat edges on LB medium; and smooth, moist, off-white colonies on Columbia blood agar medium. Gram staining revealed that the isolate was a Gram-negative, spore-free short bacillus. Biochemical tests showed that the isolate was positive for motility test, mannitol test, hydrogen sulfide test, methyl red test, citrate utilization test, gas production from glucose test, raffinose test, sorbitol test, and D-xylose test, but negative for phenylalanine test, gluconate test, indole test, Voges Proskauer (VP) test, urease test, lysine test, ornithine test, and adonitol test. The results of 16S rRNA gene sequencing showed that the gene sequence similarity between strain MF071743 and Citrobacter freundii was 99.0%. PCR results showed that this strain carries genes encoding the urease accessory proteins UreD, UreE, and UreF. Animal intragastric administration assays demonstrated that the strain caused loose stools in cynomolgus monkeys, and all C57BL/6 mice died within 72 h. Drug susceptibility test results indicated that the isolate was sensitive to 16 antibiotics, including ceftriaxone, amikacin, gentamicin, ceftazidime, and levofloxacin, while it exhibited resistance to 5 antibiotics, including ampicillin, cefazolin, vancomycin, erythromycin, and cephalexin. ConclusionA strain of Citrobacter freundii with certain drug resistance was isolated from the feces of diarrheal cynomolgus monkeys. The results of this study provide a reference for the diagnosis and treatment of gastrointestinal diseases in captive laboratory monkeys.

Objective This study conducted molecular biological identification of the viruses carried by Culex pipiens pallens specimens collected in Shunyi District, Beijing in 2023, and observed the changes in the infection rate of the viruses carried by Cx. pipiens pallens at different collection times. Methods Cx. pipiens pallens were collected using carbon dioxide mosquito traps. The mosquito samples were ground in batches and analyzed by molecular biology technologies. The virus infection rate at different collection times was analyzed statistically. Results 17 strains of Culex pipiens pallens densovirus(CppDNV)were identified from Cx. pipiens pallens samples collected in Shunyi District, Beijing, in 2023. The nucleotide sequence analysis of the virus genome coding region showed that CppDNV was a single-stranded DNA virus with a total length of 3 335 nt, encoding 2 non-structural proteins(NS1, NS2)and 1 capsid protein(VP). The nucleotide(amino acid)sequence lengths of the three proteins were 2 376 nt(791 aa),1 092 nt(363 aa)and 1 071 nt(356 aa), respectively. Phylogenetic analysis showed that CppDNV was located in genus Brevihamaparvovirus. Statistical analysis showed no significant difference in infection rates across collection times(χ2=4.429, P=0.194). Conclusions CppDNV was identified in Cx. pipiens pallens in Beijing, and it was stably maintained in this natural population.

AIM:To investigate the current status of astigmatism and its associated factors among children and adolescents from kindergarten through senior high school in Yangpu District, Shanghai, and to provide scientific evidence for the prevention and control of refractive errors.METHODS:A cluster sampling method was employed. From September to November 2024, students from 14 randomly selected schools in Yangpu District, Shanghai, were enrolled as study subjects. Autorefraction and ocular biometry were performed under non-cycloplegic conditions, and questionnaire surveys were administered. Comparisons of astigmatism degree, detection rate, and axis were made across age groups, educational stages, and sexes. Multiple linear and logistic regression analyses were conducted to identify factors associated with astigmatism.RESULTS:A total of 7 233 students aged 4-18 y were enrolled including 3 731 males, 3 502 females, with the mean age 10.13±4.18 y, astigmatism -0.76±0.82 D, and the detection rate(≤-1.00 D)was 29.23%. Both the degree and detection rate of astigmatism were correlated with age and educational stage, peaking in senior high school, with the most pronounced changes observed during primary school. Besides age, astigmatic refractive power was influenced by spherical equivalent(β=0.105, P<0.001), mean keratometry(β=-0.118, P<0.001), sex(with boys as the reference, girls: β=0.054, P<0.001), frequent sustained near work for 30-40 min(with “almost never” as the reference, β=-0.030, P=0.038), preterm birth(β=-0.023, P=0.044), maternal myopia(β=-0.028, P=0.020), and homework completion time on rest days(β=-0.035, P=0.041). The detection rate of astigmatism was associated with maternal myopia(OR=1.130, P=0.039), male sex(OR=0.773, P<0.001), shorter sleep duration on school days(OR=0.909, P=0.028), shorter sleep duration on rest days(OR=0.937, P=0.046), preterm birth(OR=1.382, P=0.012), frequent excessive head lowering(OR=1.216, P=0.045), homework completion time on rest days(OR=1.043, P=0.046), higher spherical equivalent(OR=0.891, P<0.001), and higher mean keratometry(OR=1.188, P<0.001).CONCLUSION:The degree and detection rate of astigmatism among children and adolescents from kindergarten to senior high school in Yangpu District, Shanghai are correlated with educational stage, and primary school may represent a critical period for astigmatism prevention and control. Male sex, maternal myopia, preterm birth, high-intensity near work, and insufficient sleep and other factors may be associated with the onset and progression of astigmatism. It is recommended that education and health departments strengthen visual health screening during primary school, while promoting behavioral interventions such as scientific eye-use habits and adequate sleep across different educational stages to delay the onset and progression of astigmatism.

OBJECTIVE To investigate the inhibitory effect of human β defensin-3(HBD3)on the replication of in-fluenza A virus(IAV)[A/PR/8/34(H1N1)virus strain]in human bronchial epithelial cells(BEAS-2B)via the mitochondrial autophagy pathway.METHODS BAS-2B cells were infected with IAV,and cell condition observa-tion:plaque assay and light microscopy.Drug treatment:HBD3,autophagy agonist rapamycin(Rapa),autoph-agy inhibitor LY294002;The expression levels of TUFM、MAVS、NP、M2 and PB1-F2 genes were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR).Western blot was used to detect the protein expression levels of P62、LC3Ⅱ and LC3Ⅰ.RESULTS Plaque formation experiments showed that the number of plaques increased with the increase of viral titer.With the increase of viral titer or the prolongation of infection time,the expres-sion levels of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2 B cells gradually increased,the expression levels of P62 protein decreased,and the protein expression levels of LC3Ⅱ/LC3Ⅰ increased(P＜0.05).Forty-eight hpi of BE-AS-2B cells with IAV,the number of cells in the IAV group was significantly lower than that in the IAV+HBD3 group,the intercellular space was enlarged,and the cells shrunk significantly.Compared with the IAV group,the expression lev-els of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2B cells in the IAV+HBD3 group decreased,the expres-sion levels of P62 protein increased,and the protein levels of LC3Ⅱ/LC3Ⅰ decreased(P＜0.05).Gene expression levels of TUFM,MAVS,NP,M2 and PB1-F2 were in the IAV+Rapa+HBD3 group were lower than those in the IAV+Ra-pa group(P＜0.05).The protein expression level of P62 in the IAV+Rapa+HBD3 group was higher than that in the IAV+Rapa group(P＜0.05).The protein expression levels of LC3Ⅱ/LC3Ⅰ were lower in the IAV+Rapa+HBD3 group than in the IAV+Rapa group(P＜0.05).CONCLUSION With the increase of viral titer or the prolongation of in-fection time,the proliferation of IAV in BEAS-2B cells increases,and the cell damage exacerbates.HBD3 can inhibit the replication of IAV after its entry into the cells;HBD3 can protect host cells and inhibit IAV replication by inhibiting MA-VS,TUFM-mediated mitophagy pathways.

Objective:To describe the current status of responsive caregiving behavior of infant mothers,to analyze their influencing factors and pathways using the information-motivation-behavioral skills(IMB)model,and to provide a basis for further interventions related to responsive caregiving be-haviors and comprehensive promotion of early childhood development.Methods:This study was a cross-sectional survey using convenience sampling.Questionnaires were used to collect basic information about mothers and their infants,as well as data on mothers' responsive caregiving behavior,knowledge of re-sponsive caregiving,social support,and parenting self-efficacy.Multivariate linear regression models were employed to analyze the influencing factors of responsive caregiving behavior,and structural equa-tion modeling was used to analyze the pathways of these influencing factors.The criterion for inadequate responsive caregiving is defined as scores not exceeding the lower quartile(P25)of the total score.Results:Among 510 mothers of aged 0-12 months infants in Weifang City,the average score for respon-sive caregiving behavior was 16.41±3.99.The proportion of inadequate responsive caregiving was 25.7％.Mothers in the insufficient responsive caregiving group had lower scores in knowledge(7.70±1.41),social support(57.92±15.16),and parenting self-efficacy(30.36±6.48)compared with those in the sufficient group,with statistically significant differences(P＜0.001).Logistic regres-sion analysis indicated that the influencing factors for responsive caregiving included the level of know-ledge about responsive parenting[adjusted OR(aOR)=0.795,95％CI:0.566-0.838],social support(aOR=0.979,95％CI:0.961-0.996),and parenting self-efficacy(aOR=0.894,95％CI:0.857-0.932).Structural equation modeling revealed that knowledge of responsive caregiving(β=0.089,P=0.031),social support(β=0.153,P=0.001),and parenting self-efficacy(β=0.296,P＜0.001)were directly related to responsive caregiving behavior.Additionally,knowledge of responsive caregiving indirectly affected responsive caregiving behavior through parenting self-efficacy(β=0.095,P=0.014),and social support indirectly affected responsive caregiving behavior through parenting self-efficacy(β=0.497,P＜0.001).Conclusion:The current level of responsive caregiving behavior among mothers of 0-1-year-old infants in Weifang City is not satisfactory.Future development of responsive care-giving interventions should focus on providing caregivers with relevant knowledge of responsive caregiving based on their needs.Additionally,it is essential to offer social support from multiple aspects to enhance caregivers' parenting self-efficacy,thereby promoting improvements in responsive caregiving behavior.

Objective To explore the effect and mechanism of Slc1a2 overexpression on cognitive dysfunction in sleep-deprived mice.Methods A total of 130 mice were divided into five groups:normal sleep(NS),NS+ov-Slc1a2,sleep deprivation(SD),SD+ov-NC,and SD+ov-Slc1a2,with 26 mice in each group.The SD mice model was established using an automatic system based on a rotating rod,and overexpress Slc1a2 adenovirus was injected into the prefrontal cortex(PFC).Immunofluorescence and Western blotting were used to detect the expression of Slc1a2 in the mouse PFC.Electrophysiological tests were used to evaluate non-rapid eye movement(NREM)sleep time,rapid eye movement(REM)sleep time,and wakefulness time in mice.Real-time quantitative PCR was used to detect the expression of glutamate(Glu)and gamma-aminobutyric acid(GABA)metabolic enzymes in the mouse PFC.Whole-cell patch-clamp recording was used to detect the frequency and amplitude of miniature excitatory postsynaptic currents(mEPSC)in mouse PFC.Immunofluorescence was used to detect the proportion of GABA-positive cells in the mouse PFC.The C11-BODIPY fluorescent probe was used to detect lipid reac-tive oxygen species(ROS)levels in mouse PFC.Commercial kits were used to detect Fe2+and malondialdehyde(MDA)levels in the mouse PFC.Cognitive function in mice was evaluated using the open field,novel object recognition,and Y-maze tests.Results Compared with the NS group,the NREM sleep time,REM sleep time,central area stay time,recognition index,and novel wall selection index increased significantly,while wakefulness time decreased significantly in the NS+ov-Slc1a2 group(all P＜0.05).The percentage of Slc1a2+GFAP+cells,expression of Slc1a2 protein,expression of Glul,Slc6a1,and Abat mRNA,frequency and amplitude of mEPSC,and proportion of GABA-positive cells in the PFC increased significantly,whereas lipid ROS,Fe2+,and MDA levels decreased significantly(all P＜0.05).Compared with the NS group,the NREM sleep time,REM sleep time,central area stay time,recognition index,and novel wall selection index of the SD group and the SD+ov-NC group were significantly decreased,whereas wakefulness time was significantly increased(all P＜0.05).The percentage of Slc1a2+GFAP+cells,expression of Slc1a2 protein,expression of Glul,Slc6a1,and Abat mRNA,frequency and amplitude of mEPSC,and proportion of GABA-positive cells in the mouse PFC decreased significantly,whereas lipid ROS,Fe2+,and MDA levels increased significantly(all P＜0.05).Compared with the SD and SD+ov-NC groups,the NREM sleep time,REM sleep time,central area stay time,recognition index,and novel wall selection index of the SD+ov-Slc1a2 group increased significantly,whereas the wakeful-ness time decreased significantly(all P＜0.05).The percentage of Slc1a2+GFAP+cells,the expression of Slc1a2 protein,the expression of Glul,Slc6a1,and Abat mRNA,the frequency and amplitude of mEPSC,and the proportion of GABA-positive cells in the mouse PFC increased significantly,whereas lipid ROS,Fe2+,and MDA levels decreased significantly(all P＜0.05).Conclusion Ectopic overexpres-sion of Slc1a2 in the PFC can improve sleep disorders in SD mice,reduce the damage caused by SD to excitatory synaptic transmission and GABAergic neuron function in the PFC,and alleviate cognitive impairment and anxiety-like behavior in these mice.Its mechanism may be related to the improvement of Glu/GABA metabolic imbalance in the PFC and inhibition of ferroptosis.

Objective:To investigate the MRI characteristics of ductal lesions with microcalcifications detected by mammography,and to evaluate the value of MRI in the differential diagnosis of benign and malignant ductal lesions.Methods:The clinical data of 112 patients with pathologically confirmed ductal lesions with microcalcifications detected by mammography in our hospital from January 2022 to June 2024 were retrospectively analyzed.All patients underwent mammography and MRI examinations.MRI morphological features,dynamic enhancement characteristics,diffusion-weighted imaging(DWI)findings,and apparent diffusion coefficient(ADC)values were analyzed and compared with pathological results.The diagnostic performance of various MRI features was analyzed using ROC curves.Results:Among the 112 patients,there were 47 benign lesions and 65 malignant lesions.Malignant lesions were more likely to present with irregular morphology,heterogeneous T2 signal,lobulation,and spiculation(P＜0.05),while benign lesions were more likely to present with round or oval shapes and homogeneous T2 signal(P＜0.05).Malignant lesions predominantly showed fast rise-fast washout(Washout type)enhancement curves(58.5％),while benign lesions predominantly showed persistent rise(Progressive type)enhancement curves(55.3％).The ADC values of malignant lesions(0.92±0.21× 10-3 mm2/s)were significantly lower than those of benign lesions(1.45±0.28× 10-3 mm2/s)(P＜0.001).The combined application of morphological features,dynamic enhancement curves,and ADC values had the highest diagnostic performance,with a sensitivity of 92.3％,specificity of 89.4％,and accuracy of 91.1％.Conclusion:Multi-parameter MRI can comprehensively evaluate ductal lesions with microcalcifications detected by mammography.The combined application of morphological features,dynamic enhancement curve types,and ADC values can effectively improve the accuracy of differential diagnosis between benign and malignant lesions,providing important basis for clinical treatment decisions.

Objective:To investigate the completion rate of tumor evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients in the national multicenter real-world database.Methods:The prospective real-world study was conducted. The clinicopathological data of 1 074 patients who underwent surgical treatment for mid and low rectal cancer in 47 national medical institutions, including Peking Union Medical College Hospital et al, from May 12,2023 to May 11,2024 were collected. Observation indicators: (1) clinical characteristics of patients with mid and low rectal cancer; (2) initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer; (3) initial imaging evaluation of patients with mid and low rectal cancer; (4) imaging evaluation after neoadjuvant therapy for patients with mid and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3). Count data were described as absoluter numbers and/or percentages. Results:(1) Clinical characteristics of patients with mid and low rectal cancer. Of the 1 074 patients, there were 713 males and 361 females, aged 63(56,70)years. The body mass index of 1 074 patients was 24(21,26)kg/m 2.For American Society of Anesthesiologists classification, there were 147 cases of stage Ⅰ, 641 cases of stage Ⅱ, 157 cases of stage Ⅲ, 2 cases of stage Ⅳ, and there were 127 cases missing data. (2) Initial colonoscopy and pathologic evaluation of tumors in patients with mid and low rectal cancer. Of the 1 074 patients, there were 787 cases (73.28%) undergoing complete colonoscopy, and there were only 197 cases (18.34%) undergoing immunohistochemical evaluation of all four mismatch repair proteins. (3) Initial imaging evaluation of patients with mid and low rectal cancer. Of the 1 074 patients, there were 842(78.40%) patients completing magnetic resonance imaging (MRI) or ultrasound evaluation, and there were 914(85.10%) patients completing chest, abdomen, and pelvis enhanced computed tomography (CT) evaluation. In the 149 patients completing rectal ultrasound evaluation, there were 122 cases (81.88%) comple-ting T staging evaluation, and there were 81 cases (54.36%) completing N staging evaluation. In the 808 patients completing rectal MRI evaluation, there were 708 cases (87.62%) completing T staging evaluation, and there were 590 cases (73.02%) completing N staging evaluation. (4) Imaging evalua-tion after neoadjuvant therapy for patients with mid and low rectal cancer. Of the 388 patients with neoadjuvant therapy, there were 332 patients (85.57%) completing MRI or ultrasound evaluation, and there were 327 patients (84.28%) completing chest, abdomen, and pelvis enhanced CT evalua-tion. In the 70 patients completing rectal ultrasound evaluation, there were 65 cases (92.86%) com-pleting T staging evaluation, and there were 49 cases (70.00%) completing N staging evaluation. In the 327 patients completing rectal MRI evaluation, there were 246 cases (75.23%) completing T staging, and there were 228 cases (69.72%) completing N staging evaluation. Conclusion:The com-pletion rate of tumor imaging evaluation at initial assessment and after neoadjuvant therapy for mid and low rectal cancer patients on a national scale is relatively good.

Peripheral nerve injury refers to the impairment that occurs when the sens-ory nerves and motor nerves are damaged.Professor Zhang Hong has unique insights into the treatment of neurologically diseases in children.Based on the different manifestations of the disease,it is categorized under the traditional Chinese medicine concepts of"atro-phy syndrome","muscle injury"and"Bi syndrome".Professor Zhang believes that the disease is fundamentally rooted in the liver and kidneys,and that treatment should notonly address the acquired symptoms but also nurture the innate foundation.This article a-nalyze and summarize professor Zhang's clinical experience in using acupuncture,electroa-cupuncture,moving cupping,and moxibustion in the comprehensive treatment of peripheral nerve injury,providing a reference for clinical treatment of peripheral neuropathy in chil-dren.