Objective To summarize and analyze the efficacy and influencing factors of second allogeneic hematopoietic stem cell transplantation (allo-HSCT) for acute leukemia relapsing after the first allo-HSCT. Methods Clinical data of 17 patients with acute leukemia who underwent second allo-HSCT at Peking University First Hospital from January 2005 to December 2024 were retrospectively analyzed. Results Among the 17 patients, 7 achieved long-term disease-free survival after second transplantation. The median progression-free survival after successful second transplantation was 7 months (range 8 days to 69 months). The relapse fatality was 24%, and the transplant-related fatality was 35%. Conclusions Second transplantation is an effective treatment for relapsed and refractory acute leukemia, but the relapse fatality and transplant-related fatality remain high. Patient age, time of relapse after the first transplantation and disease status before second transplantation are all factors that affect the efficacy of second transplantation. Younger age, late relapse and complete remission of disease before second transplantation are all beneficial for long-term disease-free survival after second transplantation.

Objective: To develop a prediction model for mild cognitive impairment (MCI) risk among the elderly, so as to provide a tool for MCI early screening. Methods : From July 2022 to September 2024, a multi-stage stratified random cluster sampling method was used to recruit permanent residents aged ≥65 years from the Xinjiang Uygur Autonomous Region as study participants. Data on sociodemographic characteristics, nutritional status, body composition indices, bone mineral density, and handgrip strength were collected through questionnaires and physical examinations. Sarcopenia was defined based on appendicular skeletal muscle index and handgrip strength. MCI was assessed using the Mini-Mental State Examination, with adjustments for educational level. Participants were randomly divided into a training set and a validation set in a 7∶3 ratio. LASSO regression and multivariable logistic regression models were employed to screen for predictors and construct an MCI risk prediction model. The predictive performance of the model was evaluated using receiver operating characteristic (ROC) curve and decision curve analysis (DCA). Results: A total of 1 641 participants were surveyed, including 755 males (46.01%) and 886 females (53.99%). The majority of participants were aged 65-<75 years, comprising 1 154 individuals (70.32%). MCI was detected in 517 participants, corresponding to a detection rate of 31.51%. Resultsfrom LASSO regression and multivariate logistic regression analysis showed that residence (rural, OR = 2.323, 95% CI: 1.682-3.210), age (75-<85 years, OR = 1.405, 95% CI: 1.019-1.937; ≥85 years, OR = 3.655, 95% CI: 1.696-7.875), educational level (primary school, OR = 0.341, 95% CI: 0.247-0.472; junior high school, OR = 0.255, 95% CI: 0.160-0.408; high school, OR = 0.286, 95% CI: 0.154-0.531; bachelor's degree or above, OR = 0.120, 95% CI: 0.041-0.351), history of alcohol consumption (yes, OR = 3.216, 95% CI: 2.164-4.779), risk of malnutrition (yes, OR = 1.464, 95% CI: 1.064-2.014), sarcopenia (yes, OR = 3.197, 95% CI: 2.332-4.385), and waist-to-hip ratio (abnormal, OR = 1.540, 95% CI: 1.159-2.048) were identified as predictive factors for MCI among the elderly. In the training set, the area under the ROC curve, sensitivity, and specificity were 0.788, 0.719, and 0.712, respectively. In the validation set, the corresponding values were 0.784, 0.913, and 0.542, respectively. DCA demonstrated that the model provided a higher clinical net benefit for predicting MCI risk when the risk threshold probability ranged from 0.124 to 0.764. Conclusion The prediction model developed in this study demonstrates good discriminative ability and clinical utility, indicating its substantial value for predicting the MCI risk among the elderly.

OBJECTIVE To investigate the reversal effect of Qingfei huayu jiedu formula （QFHYJDF） on drug resistance in non-small cell lung cancer （NSCLC） cells based on the microRNA-641 （miR-641）/extracellular signal-regulated kinase （ERK） signaling pathway. METHODS The human parental NSCLC cell line A549 was used to establish gefitinib-resistant cells A549/GR by combining concentration gradient increment with high-concentration pulse strategy. After screening for optimal intervention concentration and duration， the resistant cells were divided into a control serum group （treated with 20% blank serum for 48 h）， QFHYJDF-containing serum group （treated with 20% QFHYJDF-containing serum for 48 h）， and the normally cultured group （normally cultured for 48 h）. The cell relative viability， total apoptosis rate， as well as the expression levels of miR-641， neurofibromin 1 （NF1）， ERK1/2， and phosphorylated ERK1/2 （p-ERK1/2）， were assessed in each group. RESULTS Compared with the normally cultured group and the control serum group， the QFHYJDF-containing serum group exhibited significantly decreased cell relative viability and significantly increased total apoptosis rate （ P ＜0.05）. Moreover， the expression of NF1 protein was significantly up-regulated， while the expression levels of miR-641 and p-ERK1， p-ERK2 proteins were significantly down-regulated in the QFHYJDF-containing serum group （ P ＜0.05）. CONCLUSIONS QFHYJDF can inhibit proliferation and promote apoptosis of gefitinib-resistant lung cancer cells， which may be associated with up-regulating NF1 protein expression， down-regulating miR-641 expression， and inhibiting the activity of the ERK signaling pathway.

Objective To investigate the protective effect of Naoluoxintong(NLXT)on mouse hippocampal neuronal HT22 cells in the oxygen-glucose deprivation/reoxygenation(OGD/R)model,focusing on its impact on mitochondrial function and the role of the cal-cium/calmodulin-dependent protein kinase kinase β(CaMKKβ)in it.Methods HT22 cell were cultured to establish OGD/R model,and the cell were divided into normal group(Control),model group(OGD/R),NLXT group(OGD/R+10％NLXT),positive drug dl-3-n-butylphthalide(NBP)group(OGD/R+NBP 50μmol/L)after the screening of the optimal concentrations of different con-centrations of NLXT-containing serum(0-20％)in interfering with the HT22 cells by the CCK-8 assay.Then,using CCK-8 assay to detect the cell survival rate of each group,inverted fluorescence microscope to observe the morphology of each group,flow cytometry to detect the apoptosis rate of each group,and the kit to detect the activity or content of malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH)in each group.Nest,the CaMKKβ inhibitor(STO-609,5μmol/L)was introduced,HT22 cells were di-vided into Control group,OGD/R group,OGD/R+10％NLXT group,OGD/R+NBP group,OGD/R+STO-609group,OGD/R+STO-609+NLXT group.The mitochondrial membrane potential in each group was observed by using JC-1 fluorescent probe,and the end products Na+-K+-ATP,Ca+-Mg+-ATPase activity were detected by the kit.The expression changes of CaMKKβ protein in cells of each group were detected by Western blot,the changes of mitochondrial DNA copy number and CaMKKβ mRNA expression in cells of each group were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results The optimal concentration of NLXT-containing serum was 10％,which resulted in the most significant effects.Compared with the OGD/R group,NLXT intervention significantly increased cell viability(P＜0.05)and restored cellular morphological characteristics.Compared with the Control group,the OGD/R group showed significantly increased apoptosis(P＜0.05),elevated MDA levels,and decreased activities of SOD and GSH,alongside a reduction in mitochondrial membrane potential,ATP activity,and mitochondrial DNA copy number.Further-more,CaMKKβ protein and mRNA expression were downregulated(P＜0.05).Compared with the OGD/R group,the NLXT and NBP groups antagonized these effects and upregulated CaMKKβ protein and mRNA expression(P＜0.05).Additionally,the application of the CaMKKβ inhibitor STO-609 significantly attenuated the protective effects of NLXT on mitochondrial function and neurons(P＜0.05).Conclusion NLXT exerts neuroprotective effects by activating the CaMKKβ to enhance mitochondrial function after OGD/R in HT22 cells.

AIM:To explore the mechanism of berberine on breast cancer cells based on network pharmacology and in vitro cell experiments.METH-ODS:Firstly,berberine and breast cancer were tak-en as the research objects,the intersection targets of the two were screened by VEEN diagram,GO function and KEGG enrichment analysis were per-formed by R language,and molecular docking and visualization were carried out by Autodock Vina and Pymol software.Then,berberine treated breast cancer MCF-7 cells for 24 h,and then in vi-tro cell experiments were performed.CCK-8 was used to detect cell viability,Edu and plate cloning were used to detect cell proliferation and cloning,and apoptosis was detected by An-nexin V-FITC/PI double staining and Western blot.Laser confocal and CETSA were used to verify the binding effect of berberine and AKT1 protein.RESULTS:The results of network pharmacology showed that berberine had a good binding to the core targets AKT1,AKT2 and MAPK3.Berberine(20,40,80 μmol/L)signifi-cantly inhibited the proliferation and cloning ability of MCF-7 cells in a concentration-dependent man-ner(P＜0.05,P＜0.01).The results of laser confocal and CETSA experiments showed that berberine and AKT1 had a binding effect,and the stability of the two was enhanced after the combination.CONCLU-SION:Berberine inhibits MCF-7 cell proliferation and induces apoptosis in human breast cancer cells by targeting binding to AKT1 protein.

OBJECTIVE To explore the molecular characteristics of small colony variants(SCVs)of Staphylococcus aureus isolates so as to provide theoretical bases for clinical control of the persistent and recurrent infections in-duced by the SCVs.METHODS The clinical blood specimens that were collected from the First Affiliated Hospital of Traditional Chinese Medicine University of Guangzhou were cultured to acquire 1 strain of SCV S.aureus S2.The colonial morphology,growth rate,catalase,plasma-coagulase,production capability of biofilm and antimi-crobial susceptibility of the strain SCVs were observed,the growth curve and autolysis curve were drawn,the whole genome sequencing was performed for the S2,the phylogenetic evolution of the genome and the single nu-cleotide polymorphism(SNP)were analyzed.RESULTS The S2 was identified as S.aureus,with the catalase and plasma-coagulase tested positive.The strain was resistant to various types of antibiotics,including macrolides like erythromycin and azithromycin,fluroquinolones such as levofloxacin and ciprofloxacin,and penicillin.The forma-tion of pigment was reduced,the formation of hemolytic rings was low;the growth rate of the strain was slower than that of ATCC 25923(quality control strain)and ZC1(the normal control strain),the autolytic activity of the strain was higher than that of the ATCC 25923 and ZC1,and the biofilm production of the strain was remarkably less than that of the ATCC 29213(P＜0.05).The whole genome sequencing demonstrated that the molecular typ-ing of the S2 was ST239-t030 clone,which harbored a variety of drug resistance genes and virulence genes,and the menB gene had missense mutation.CONCLUSION The formation of SCVs may be associated with the inhibited synthesis of menadione caused by the missense mutation of menB gene.

Objective:To observe whether electroacupuncture(EA)can inhibit activation of JAK1/STAT3 signaling pathway and improve pulmonary inflammation in chronic obstructive pulmonary disease(COPD)by increasing expression of SOCS3.Methods:Atotal of 60 mice were randomly divided into normal group,model(COPD)group,COPD+EA group,COPD+si-SOCS3 group,COPD+si-SOCS3 NC group,COPD+si-SOCS3+EA group,with 10 mice in each group.Mice COPD model was replicated by simple cigarette smoking for three months.After modeling,SOCS3 siRNA was administered to lungs of mice in COPD+si-SOCS3 group and COPD+si-SOCS3+EA group,SOCS3 siRNA NC was administered to COPD+si-SOCS3 NC group.24 h after the first SOCS3 siRNA ad-ministration,mice in COPD+EA and COPD+si-SOCS3+EA groups were treated with EA in"Feishu"and"Zusanli",once every other day,30 min/times for 14 days.Lung function of mice in each group was detected;lung pathological changes were observed by HE staining;IL-6,IL-1β and TNF-α levels in mice broncho alveolar lavage fluid were detected by ELISA;protein expressions of SOCS3,JAK1,STAT3,p-JAK1 and p-STAT3 were detected by Western blot;SOCS3,JAK1 and STAT3 mRNA were detected by RT-PCR.Results:Compared with normal group,mice in COPD group showed decreased lung function,thickened alveolar walls,congestion and edema between tissues,significantly increased levels of inflammatory cytokines IL-6,TNF-α and IL-1β,decreased SOCS3 protein expres-sion,p-JAK1/JAK1 and p-STAT3/STAT3 significantly increased,JAK1 and STAT3 mRNA increased,SOCS3 mRNA decreased(P<0.05).Compared with COPD group,the above indexes of COPD+EA group were improved(P<0.05),and the above indexes of COPD+si-SOCS3 group were more serious(P<0.05),and COPD+si-SOCS3+EA group was improved compared with COPD+si-SOCS3 group(P<0.05).Conclusion:EA can inhibit overactivation of JAK1/STAT3 signaling pathway by up-regulating SOCS3 expression,and thus improve pulmonary inflammation in COPD.

Objective To investigate the predictive value of bone metabolism parameters on cardiac autonomic nervous system function in patients with type 2 diabetes mellitus(T2DM).Methods A total of 328 patients with T2DM hospitalized in the Department of Endocrinology of Gansu Provincial People's Hospital were enrolled in this study from October 2022 to October 2023.According to the serum 25(OH)D level,all the participants were divided into<10 ng/ml group(n=80),10～20 ng/ml group(n=173),and 20～30 ng/ml group(n=75).Biochemical indicators,bone metabolic parameters,left ventricular mass(LVM)and left ventricular mass index(LVMI)were compared.Time domain indicators ofheart rate variability(HRV)in 24 h holter electrocardiogram,including the global standard deviation of normal sinus RR interval(SDNN),sinus RR interval mean standard deviation(SDANN),and normal continuous sinus RR interval difference root mean square(RMSSD).Meanwhile,adjacent RR interval difference>50 ms as a percentage of the total inter-period(PNN50),HRV triangle index,standard deviation of the difference between the length of the entire adjacent NN interperiod(SDSD),and 24 h holter electrocardiogram HRV time-domain relevant indicators were compared among the three groups.The influence of bone metabolism parameters on cardiac autonomic nervous function and their correlation were analyzed,and the optimal cutting point of cardiac autonomic nervous function was predicted by receiver operating characteristic(ROC)curve.Results SBP,heart rate(HR),FPG,PWV,PTH and β-CTX in groups of 10 ng/ml,10～20 ng/ml and 20～30 ng/ml decreased in turn(P<0.05),while HDL-C,ABI,25(OH)D,Ca2+and PNN50 decreased.Correlation analysis between Spearman and Pearson showed that 25(OH)D was positively correlated with SDNN,HRV triangle index,PNN50 and rMSSD(P<0.01).Logistic regression analysis showed that 25(OH)D,Ca2+and HR were the influencing factors of cardiac autonomic nervous dysfunction in patients with T2DM.The ROC curve analysis showed that the areas under the ROC curve of 25(OH)D,Ca2+and HR were 0.791,0.607 and 0.629,respectively,with sensitivity of 73.4%,53.2%and 38.7%,and specificity of 74.0%,93.6%and 81.4%,respectively.Conclusions 25(OH)D is the influencing factor of cardiac autonomic nervous dysfunction in patients with T2DM,and patients with high degree of deficiency are more prone to cardiac autonomic nervous dysfunction.

Objective To investigate the protective effect of Naoluoxintong(NLXT)on mouse hippocampal neuronal HT22 cells in the oxygen-glucose deprivation/reoxygenation(OGD/R)model,focusing on its impact on mitochondrial function and the role of the cal-cium/calmodulin-dependent protein kinase kinase β(CaMKKβ)in it.Methods HT22 cell were cultured to establish OGD/R model,and the cell were divided into normal group(Control),model group(OGD/R),NLXT group(OGD/R+10％NLXT),positive drug dl-3-n-butylphthalide(NBP)group(OGD/R+NBP 50μmol/L)after the screening of the optimal concentrations of different con-centrations of NLXT-containing serum(0-20％)in interfering with the HT22 cells by the CCK-8 assay.Then,using CCK-8 assay to detect the cell survival rate of each group,inverted fluorescence microscope to observe the morphology of each group,flow cytometry to detect the apoptosis rate of each group,and the kit to detect the activity or content of malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH)in each group.Nest,the CaMKKβ inhibitor(STO-609,5μmol/L)was introduced,HT22 cells were di-vided into Control group,OGD/R group,OGD/R+10％NLXT group,OGD/R+NBP group,OGD/R+STO-609group,OGD/R+STO-609+NLXT group.The mitochondrial membrane potential in each group was observed by using JC-1 fluorescent probe,and the end products Na+-K+-ATP,Ca+-Mg+-ATPase activity were detected by the kit.The expression changes of CaMKKβ protein in cells of each group were detected by Western blot,the changes of mitochondrial DNA copy number and CaMKKβ mRNA expression in cells of each group were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results The optimal concentration of NLXT-containing serum was 10％,which resulted in the most significant effects.Compared with the OGD/R group,NLXT intervention significantly increased cell viability(P＜0.05)and restored cellular morphological characteristics.Compared with the Control group,the OGD/R group showed significantly increased apoptosis(P＜0.05),elevated MDA levels,and decreased activities of SOD and GSH,alongside a reduction in mitochondrial membrane potential,ATP activity,and mitochondrial DNA copy number.Further-more,CaMKKβ protein and mRNA expression were downregulated(P＜0.05).Compared with the OGD/R group,the NLXT and NBP groups antagonized these effects and upregulated CaMKKβ protein and mRNA expression(P＜0.05).Additionally,the application of the CaMKKβ inhibitor STO-609 significantly attenuated the protective effects of NLXT on mitochondrial function and neurons(P＜0.05).Conclusion NLXT exerts neuroprotective effects by activating the CaMKKβ to enhance mitochondrial function after OGD/R in HT22 cells.

Objective:To investigate the association between high-risk human papillomavirus (hrHPV) persistent infection and vaginal microecology.Methods:A total of 53 women were enrolled in the gynecological clinic of Beijing Obstetrics and Gynecology Hospital from January 2020 to January 2021, including 7 women without HPV and 46 women with hrHPV infection. Among the hrHPV infected women, 24 woemn who did not use any drugs were classified as the observation group and the other 22 women who were given standardized interferon vaginal administration for 3 months were regarded as the treatment group. Vaginal secretions of all women were taken for Gram-stained microecological test at the time of enrollment and at the 4, 8, and 12 month follow-up. HPV turning negative was taken as the end point of follow-up.Results:(1) Women of hrHPV persistent infection in the observation and treatmnet groups had more times of abortions ( P=0.180). (2) The hrHPV negative conversion rate was 17% (4/24) in the observation group and 36% (8/22) in the treatment group, but the difference was not significant ( P=0.183). The median hrHPV negative conversion time were 11.0 months and 7.5 months in the observation and treatment groups, respectively, and the difference was statistically significant ( P=0.001). (3) Vaginal microecology was generally normal at the time of enrollment and at the end of follow-up in women with HPV natural negative conversion in the observation group. While vaginal microecological disorders were more common in women with hrHPV persistent infection in the observation and treatmnet groups, including high vaginal pH value, poor vaginal cleanliness, poor grade of Lactobacillus and increased vaginal clutter bacteria, and the vaginal microecological situation did not improve after the 12-month follow-up. (4) In the treatment group, women who turned HPV negative within six months all had normal vaginal microecology when enrollment (5/5). While those who turned negative six months later had a higher proportion of vaginal clutter bacteria (2/3), a poor grade of Lactobacillus (2/3) and a higher proportion of vaginal dysbiosis (2/3). Conclusions:(1) Interferon therapy could shorten the negative turning time of hrHPV. (2) Women with normal vaginal microecology have the ability to naturally clear hrHPV. (3) The vaginal microecological Gram-stain test has limited value in predicting hrHPV clearance, perhaps due to its inability to detect Lactobacillus subtypes.