Objective:To explore the significance and value of peripheral blood lymphocyte subpopulation characteristics in patients with leukemia recurrence after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A retrospective cohort study was conducted on 115 leukemia patients with allo HSCT admitted to Beijing Ludaopei Hospital from April 1, 2021 to December 31, 2023, including 26 relapsed patients and 89 non-relapsed patients, 65 males and 50 females aged 33 (20, 50) years old. Flow cytometry was used for regular monitoring of the post-transplantation peripheral blood lymphocyte subpopulations in the patients, i.e. total lymphocytes, B-lymphocytes, T-lymphocytes, NK-cells, CD4+Treg-cells, CD3+CD4+CD8+T-cells, CD3+CD4-CD8-T-cells, CD28 T-cells. The significance of lymphocyte subpopulation detection in the relapsed group of leukemia patients was further discussed. The χ 2 test was used for comparison between the two groups. Indicators statistically significant in one-way analysis were then analyzed by binary logistic regression analysis. Results:Compared with patients in the non-relapse group, patients in the relapse group had lower total lymphocytes (458.5<1 003/μl, U=1 118.00, P=0.004), B lymphocytes (4<51/μl, U=67.00, P<0.001), T lymphocytes (320.5<721/μl, U=746.00, P=0.006), CD4+T lymphocytes (57<150/μl, U=629.50, P<0.001), CD8+T lymphocytes (240<468/μl, U=1 150.00, P=0.017), CD3+CD4-CD8-T cells (5<32.5/μl, U=259.00, P<0.001), CD8+CD28+T cells (92<298/μl, U=294.00, P=0.002), CD4+CD28+T cell counts (43<141/μl, U=245.50, P<0.001), CD8+CD28+T cells as a percentage of CD8+T cells (63.3<70.9%, U=331.00, P=0.006), and CD4+CD28+T cells as a percentage of CD4+T cells (86.1<97.3%, U=313.50, P=0.003), while the CD4+Treg cell count (2.5>8/μl, U=635.00, P<0.001) was higher. The percentage of CD8+CD28+T cells to CD8+T cells in the peripheral blood of patients after transplantation was a significant independent risk factor for relapse, and the results of the ROC curve showed an AUC of 0.724 ( P<0.01), with a decision value of 66.89, a sensitivity of 0.733, and a specificity of 0.613. Conclusions:The low percentage of CD8+CD28+T cells to CD8+T cells in peripheral blood has a certain value of suggestive warning for post-transplantation leukemia relapse. The application of flow cytometry for regular monitoring of patients′ peripheral blood immune cell subpopulations after transplantation helps to understand patients′ immune reconstitution status and provides a certain reference basis for clinical diagnosis and treatment.

To investigate the in vitro inhibitory mechanism of Nymphaea candida total flavonoids(NCTF)against Staphylococcus aureus(S.aureus)and its safety in mice,this study first deter-mined the antibacterial effect of NCTF on the clinically isolated strain S.aureus-C1.Subsequently,the inhibitory mechanism of NCTF on S.aureus-C1 was explored by measuring its effects on bac-terial growth curves,microstructure,intracellular AKP and LDH levels,and biofilm formation.Safety evaluation included determination of LD50 and MDT in mice,as well as analysis of serum biochemical parameters,organ indices,and histopathological observations.Results showed that NCTF effectively inhibited S.aureus-C1 proliferation,with an inhibition zone diameter of(18.98±0.67)mm and a MIC of 6.25 g/L.A concentration of 2×MIC nearly completely suppressed bacte-rial growth.Scanning electron microscopy revealed structural damage to bacterial cells,including collapse and shrinkage.AKP and LDH assays indicated significantly increased AKP activity(P＜0.05)and decreased intracellular LDH activity(P＜0.05)in the supernatant of drug-treated groups,demonstrating NCTF-induced disruption of cell walls and membranes leading to leakage of AKP and LDH.Crystal violet staining of biofilms showed significant inhibition rates of(43.77±9.16)％and(61.71±9.82)％at 2 × MIC and 4 × MIC concentrations,respectively(P＜0.05).Safe-ty assessments indicated low toxicity of NCTF in mice,with transient effects that returned to nor-mal levels within a short period.These findings demonstrate that NCTF exhibits potent antibacte-rial activity against S.aureus-C1 by damaging bacterial cell structures,increasing cell wall/mem-brane permeability,reducing biofilm formation,and displaying low toxicity.This study provides scientific evidence for clinical drug screening against bovine mastitis and the development of Nym-phaea candida resources.

Objective:To investigate the protective effect of achyranthes bidentata(AB)on sperm quality in mice with sper-matogenic disorder through the glycolytic metabolic pathway and its action mechanism.Methods:We equally randomized 40 Kun-ming mice into a normal control,a model control,a low-dose AB(3.5 g/kg)and a high-dose AB group(7.0 g/kg),and established the model of spermatogenic disorder in the latter three groups of mice by intraperitoneal injection of doxorubicin(30 mg/kg).Two days after modeling,we collected the testis and kidney tissues and blood samples from the mice for observation of the pathological changes in the testis tissue by HE staining,detection of perm motility with the sperm quality analyzer,examination of the apoptosis of testis cells by flow cytometry,measurement of the levels of testosterone(T),malondialdehyde(MDA),superoxide dismutase(SOD)and cata-lase(CAT)in the serum and testis tissue by ELISA,and determination of expressions of the key enzymes of glycolysis hexokinase Ⅱ(HK2),pyruvate kinase M2(PKM2),platelet phosphofructokinase(PFKP),lactate dehydrogenase A(LDHA)and the meiosis pro-teins REC8 and SCP3 by Western blot,and the mRNA expressions of glycolytic phosphofructokinase 1(PFK1),phosphoglycerate ki-nase 1(PGK1),tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1β)by fluorescence quantitative PCR(FQ-PCR).Results:Compared with the model controls,the mice in the AB groups showed significant increases in the testis coefficient,kidney in-dex,sperm concentration,sperm motility,spermatogonia,primary spermatocytes,spermatids,sperm count and the serum T level(P＜0.05 orP＜0.01),but dramatic decreases in the apoptosis of testis cells and percentage of morphologically abnormal sperm(P＜0.01).Achyranthes bidentata also significantly elevated the levels of SOD and CAT,and down-regulated the mRNA expressions of MDA,TNF-α and IL-1β(P＜0.05 or P＜0.01),and up-regulated the protein expressions of HK2,PKM2,PFKP,LDHA,REC8 and SCP3,and expressions of the glycolysis key genes Pfk1 and Pgk1(P＜0.05 orP＜0.01).Conclusion:Achyranthes bidentata ameliorates doxorubicin-induced spermatogenic disorder in mice by regulating the glycolytic pathway and reducing oxidative stress and the expressions of inflammatory factors.

Hyssopus cuspidatus is an authentic medicinal herb used in Xinjiang， rich in the chemical constituents including flavonoids， phenolic acids and terpenoids. It possesses anti-inflammatory， antioxidant， and immunomodulatory effects. Modern pharmacological studies have demonstrated that H. cuspidatus exerts therapeutic effects on asthma by inhibiting airway inflammatory responses， suppressing airway remodeling， modulating the function of the hypothalamic-pituitary-adrenal axis， and relaxing bronchial smooth muscle. It also demonstrates intervention effects on chronic obstructive pulmonary disease by reducing levels of inflammatory cytokines and regulating immune balance. Additionally， H. cuspidatus can mitigate acute lung injury by inhibiting oxidative stress and intervene in lung cancer by suppressing proliferation and promoting apoptosis of lung cancer cells. In terms of clinical application， compound preparations containing H. cuspidatus， such as Hanchuan zupa granules and Luo’ou kezupa， have demonstrated favorable therapeutic effects on pulmonary diseases including asthma， cough， and pediatric bronchopneumonia. Currently， the research on precise action targets and compound matching rules of H. cuspidatus remains inadequate. Future studies should integrate modern technologies such as metabolomics to conduct in-depth exploration， thereby promoting the modernized development and clinical application of this traditional medicinal herb.

Objective To observe the efficacy of self-guided attention network for detecting responsible lesions related to cerebral palsy(CP)in children with periventricular white matter injury(PVWMI).Methods Totally 383 children with PVWMI were retrospectively enrolled and divided into CP group(n=243)and non-CP group(n=140),while 214 children without obvious brain abnormality on brain MRI were taken as control group.ROI of 4 key anatomical structures related to CP,i.e.centrum semiovale,posterior limb of internal capsule,cerebral peduncle and thalamus were delineated on T1WI,while responsible lesions related to CP within the key anatomical structures were labeled on T2WI,and the images were then registrated and used as input of the networks.ResNet34 network was adopted combined with attention and self-guided networks to train the network for detecting responsible lesions related to CP in children with PVWMI,and their efficacies were evaluated.The optimal network was screened,and its efficacy for segmenting the key anatomical structures was evaluated.Results Self-guided attention network was the optimal network,its area under the curve(AUC)for detecting lesions was 0.794-0.914,and the Dice similarity coefficient for segmenting the key anatomical structures was 0.702-0.764.Conclusion Self-guided attention network could be used for preliminarily detecting responsible lesions related to CP in children with PVWMI.

To investigate the in vitro inhibitory mechanism of Nymphaea candida total flavonoids(NCTF)against Staphylococcus aureus(S.aureus)and its safety in mice,this study first deter-mined the antibacterial effect of NCTF on the clinically isolated strain S.aureus-C1.Subsequently,the inhibitory mechanism of NCTF on S.aureus-C1 was explored by measuring its effects on bac-terial growth curves,microstructure,intracellular AKP and LDH levels,and biofilm formation.Safety evaluation included determination of LD50 and MDT in mice,as well as analysis of serum biochemical parameters,organ indices,and histopathological observations.Results showed that NCTF effectively inhibited S.aureus-C1 proliferation,with an inhibition zone diameter of(18.98±0.67)mm and a MIC of 6.25 g/L.A concentration of 2×MIC nearly completely suppressed bacte-rial growth.Scanning electron microscopy revealed structural damage to bacterial cells,including collapse and shrinkage.AKP and LDH assays indicated significantly increased AKP activity(P＜0.05)and decreased intracellular LDH activity(P＜0.05)in the supernatant of drug-treated groups,demonstrating NCTF-induced disruption of cell walls and membranes leading to leakage of AKP and LDH.Crystal violet staining of biofilms showed significant inhibition rates of(43.77±9.16)％and(61.71±9.82)％at 2 × MIC and 4 × MIC concentrations,respectively(P＜0.05).Safe-ty assessments indicated low toxicity of NCTF in mice,with transient effects that returned to nor-mal levels within a short period.These findings demonstrate that NCTF exhibits potent antibacte-rial activity against S.aureus-C1 by damaging bacterial cell structures,increasing cell wall/mem-brane permeability,reducing biofilm formation,and displaying low toxicity.This study provides scientific evidence for clinical drug screening against bovine mastitis and the development of Nym-phaea candida resources.

Objective To observe the efficacy of self-guided attention network for detecting responsible lesions related to cerebral palsy(CP)in children with periventricular white matter injury(PVWMI).Methods Totally 383 children with PVWMI were retrospectively enrolled and divided into CP group(n=243)and non-CP group(n=140),while 214 children without obvious brain abnormality on brain MRI were taken as control group.ROI of 4 key anatomical structures related to CP,i.e.centrum semiovale,posterior limb of internal capsule,cerebral peduncle and thalamus were delineated on T1WI,while responsible lesions related to CP within the key anatomical structures were labeled on T2WI,and the images were then registrated and used as input of the networks.ResNet34 network was adopted combined with attention and self-guided networks to train the network for detecting responsible lesions related to CP in children with PVWMI,and their efficacies were evaluated.The optimal network was screened,and its efficacy for segmenting the key anatomical structures was evaluated.Results Self-guided attention network was the optimal network,its area under the curve(AUC)for detecting lesions was 0.794-0.914,and the Dice similarity coefficient for segmenting the key anatomical structures was 0.702-0.764.Conclusion Self-guided attention network could be used for preliminarily detecting responsible lesions related to CP in children with PVWMI.

Objective:To explore the significance and value of peripheral blood lymphocyte subpopulation characteristics in patients with leukemia recurrence after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A retrospective cohort study was conducted on 115 leukemia patients with allo HSCT admitted to Beijing Ludaopei Hospital from April 1, 2021 to December 31, 2023, including 26 relapsed patients and 89 non-relapsed patients, 65 males and 50 females aged 33 (20, 50) years old. Flow cytometry was used for regular monitoring of the post-transplantation peripheral blood lymphocyte subpopulations in the patients, i.e. total lymphocytes, B-lymphocytes, T-lymphocytes, NK-cells, CD4+Treg-cells, CD3+CD4+CD8+T-cells, CD3+CD4-CD8-T-cells, CD28 T-cells. The significance of lymphocyte subpopulation detection in the relapsed group of leukemia patients was further discussed. The χ 2 test was used for comparison between the two groups. Indicators statistically significant in one-way analysis were then analyzed by binary logistic regression analysis. Results:Compared with patients in the non-relapse group, patients in the relapse group had lower total lymphocytes (458.5<1 003/μl, U=1 118.00, P=0.004), B lymphocytes (4<51/μl, U=67.00, P<0.001), T lymphocytes (320.5<721/μl, U=746.00, P=0.006), CD4+T lymphocytes (57<150/μl, U=629.50, P<0.001), CD8+T lymphocytes (240<468/μl, U=1 150.00, P=0.017), CD3+CD4-CD8-T cells (5<32.5/μl, U=259.00, P<0.001), CD8+CD28+T cells (92<298/μl, U=294.00, P=0.002), CD4+CD28+T cell counts (43<141/μl, U=245.50, P<0.001), CD8+CD28+T cells as a percentage of CD8+T cells (63.3<70.9%, U=331.00, P=0.006), and CD4+CD28+T cells as a percentage of CD4+T cells (86.1<97.3%, U=313.50, P=0.003), while the CD4+Treg cell count (2.5>8/μl, U=635.00, P<0.001) was higher. The percentage of CD8+CD28+T cells to CD8+T cells in the peripheral blood of patients after transplantation was a significant independent risk factor for relapse, and the results of the ROC curve showed an AUC of 0.724 ( P<0.01), with a decision value of 66.89, a sensitivity of 0.733, and a specificity of 0.613. Conclusions:The low percentage of CD8+CD28+T cells to CD8+T cells in peripheral blood has a certain value of suggestive warning for post-transplantation leukemia relapse. The application of flow cytometry for regular monitoring of patients′ peripheral blood immune cell subpopulations after transplantation helps to understand patients′ immune reconstitution status and provides a certain reference basis for clinical diagnosis and treatment.

Objective To explore the relationship and internal path between activities of daily living(ADL),sleep quality and mental health of community elderly people in Shanghai.Methods A questionnaire survey was conducted among community residents aged 60 years and older seeing doctors in community health care center of five streets in Shanghai during Sept to Dec,2021 using convenience sampling.Activities of Daily Living(ADL),Pittsburgh Sleep Quality Index(PSQI)and 10-item Kessler Psychological Distress Scale(K10)were adopted in the survey.Single factor analysis,correlation analysis and multiple linear regression were used to analyze the data.The effect relationship between the variables was tested using Bootstrap's mediated effects test.Results A total of 1 864 participants were included in the study.The average score was 15.53±4.47 for ADL,5.60±3.71 for PSQI and 15.50±6.28 for K10.The rate of ADL impairment,poor sleep quality,poor and very poor mental health of the elderly were 23.6%,27.3%,11.9%and 4.9%,respectively.ADL and sleep quality were all positively correlated with mental health(r=0.321,P＜0.001;r=0.466,P＜0.001);ADL was positively correlated with sleep quality(r=0.294,P＜0.001).Multiple linear results of factors influencing mental health showed that ADL(β= 0.457,95%CI:0.341-0.573),sleep quality(β =0.667,95%CI:0.598-0.737)and mental health were positively correlated(P＜0.001).Sleep quality partially mediated the relationship between ADL and mental health(95%CI:0.078-0.124)with an effect size of 33.0%.Conclusion Sleep quality is a mediator between ADL and mental health among community elderly people.Improving ADL and sleep quality may improve mental health in the population.

Objective:To investigate the significance and value of peripheral blood lymphocyte subset characteristics in patients following allogeneic hematopoietic stem cell transplantation (allo-HSCT) for distinguishing cytomegalovirus (CMV) infection.Methods:This retrospective study gathered data from allo-HSCT patients treated at Beijing Ludaopei Hospital between May 2021 and January 2023. This study included 50 patients in the CMV infection group and 47 patients in the non-CMV infection group. Flow cytometry was used to detect peripheral blood lymphocyte subsets 1 month after the allo-HSCT. The absolute lymphocyte count and their subsets were analyzed to assess their predictive significance in the occurrence of CMV infection. Mamn-whiney U test and χ 2 test were used. Indicators with statistical significance in univariate analysis were further subjected to binary logistic regression analysis and receiver operating characteristic (ROC) curves construction. The generation of survival curves for different outcomes was conducted using the Kaplan-Meier method, and the Log-rank test was applied. A two-sided P-value of<0.05 was considered statistically significant. Results:1. Among patients in the CMV infection group, 22% (11/50) were diagnosed with CMV viremia combined with CMV disease. The maximum CMV viral load in peripheral whole blood in these patients was 1 600 (1 400, 5 800) copies/ml, which was significantly higher than that in patients without confirmed CMV disease 970 (670, 2 300) copies/ml, with a statistically significant difference ( Z=-2.281, P=0.029). 2. The overall survival (OS) at the follow-up endpoint was 80% (40/50) in the CMV infection group and 87.2% (41/47) in the non-CMV infection group. There was no statistically significant difference in OS between the two groups at the follow-up endpoint ( χ2=0.231, P=0.630). 3. In the binary logistic regression model, the number of CD34 cells transfused into the patient, the percentage of CD3+CD38+T cells in CD3+T cells, and the percentage of CD4+CD38+T cells in CD4+lymphocytes were identified as important independent risk factors for infection. The ROC curve analysis demonstrated that the area under the curve (AUC) for these independent risk factors of CMV infection was 0.914 (>0.7, P<0.001), with a decision value of 0.702, a sensitivity of 0.857, and a specificity of 0.844. Conclusion:Followingallo-HSCT, the immune subsets in the CMV infection group exhibited a diminished proportion of T cells and CD4+T cells in comparison to the non-CMV infection group. In terms of T cell differentiation, there was an increase in TEM cells and a decrease in Naive T cells. Additionally, the expression level of the activation marker CD38 on T cells exhibited a high degree of predictive value for the occurrence of CMV infection. Understanding the immune characteristics of post-transplant CMV-infected patients is beneficial for the analysis of their immune reconstitution status, providing valuable insights for clinicians in the diagnosis and treatment of CMV infection after transplantation.