OBJECTIVE To investigate the mechanism by which Ginkgo flavone aglycone （GA） reduces the cardiotoxicity of doxorubicin （DOX） based on transcriptomics and proteomics. METHODS Thirty-six mice were randomly assigned to control group （CON group， tail vein injection of equal volume of physiological saline every other day+daily intragastric administration of an equal volume of physiological saline）， DOX group （tail vein injection of 3 mg/kg DOX every other day）， and GDOX group （daily intragastric administration of 100 mg/kg GA+tail vein injection of 3 mg/kg DOX every other day）， with 12 mice in each group. The administration of drugs/physiological saline was continued for 15 days. Mouse heart tissues were collected for RNA-Seq transcriptomic sequencing and 4D-Label-free quantitative proteomic analysis to screen differentially expressed genes and proteins， which were then subjected to Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis. The expression levels of Apelin peptide （Apelin）， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） mRNA and protein in mouse heart tissues， as well as the phosphorylation levels of PI3K and Akt proteins， were verified. H9c2 cardiomyocytes were divided into control group （CON group）， DOX group （2 μmol/L）， and GDOX group （2 μg/mL GA+2 μmol/L DOX） to determine cell viability and the levels of key glycolytic substances in the cells. RESULTS Six common pathways were identified from transcriptomics and proteomics， including the Apelin signaling pathway， the PI3K-Akt signaling pathway， and insulin resistance. Among them， the Apelin and PI3K-Akt signaling pathways were the most enriched in terms of gene numbers. Target validation experiments showed that compared to the CON group， the relative expression of Apelin， PI3K and Akt mRNA and protein levels， as well as the phosphorylation levels of PI3K and Akt proteins， were significantly decreased in the DOX group （P＜0.05 or P＜0.01）. The relative expression of Apelin， PI3K and Akt mRNA and the phosphorylation levels of PI3K and Akt proteins were significantly increased in the GDOX group as compared with the DOX group （P＜0.05 or P＜0.01）. Cellular experiments indicated that compared to the CON group， cell viability in the DOX group was significantly decreased （P＜0.05）， the relative uptake of glucose and the relative production of pyruvate and lactate were significantly increased （P＜0.05）， and the relative production of ATP was significantly reduced （P＜0.05）. Compared to the DOX group， cell viability in the GDOX group was significantly increased （P＜ 0.05）， and the relative production of pyruvate and lactate was significantly reduced （P＜0.05）. CONCLUSIONS GA may alleviate DOX-induced cardiotoxicity by upregulating the mRNA and protein expression of Apelin， PI3K， and Akt in heart tissues， and regulating glycolytic processes.

This study utilized a chiral liquid chromatography-mass spectrometry (LC-MS)-guided isolation strategy to accurately capture angular-type pyranocoumarins (APs) in Peucedani Radix (Chinese name: Qianhu). Sixteen APs were successfully purified from the ethyl acetate extract of Peucedani Radix through deploying various techniques such as silica gel, ODS, Sephadex LH-20, and achiral (chiral) semi-preparative liquid chromatography. After extensive structural measurements, such as ¹H and ¹³C NMR spectroscopy, their structures were identified as (3′S)-3′-(2-methyl-butyroyl)-4′-oxo-3′,4′-dihydroseselin (1A), (3′R)-3′-(2-methyl-butyroyl)-4′-oxo-3′,4′-dihydroseselin (1B), (3′S)-3′-isovaleryl-4′-oxo-lomatin (2A), (3′R)-3′-isovaleryl-4′-oxo-lomatin (2B), (3′S)-3′-angeloyloxy-4′-oxo-3′,4′-dihydroseselin (3A), (3′R)-3′-angeloyloxy-4′-oxo-3′,4′-dihydroseselin (3B), (3′S,4′S)-praeruptorin B (4A), (3′R,4′R)-praeruptorin B (4B), (3′S,4′S)-praeruptorin E (5A), (3′R,4′R)-praeruptorin E (5B), 3′-isovaleryl-4′-angeloyl-cis-khellactone (6), 3′-angeloyl-4′-(2-methyl-butyroyl)-cis-khellactone (7), (3′S,4′S)-praeruptorin A (8A), (3′R,4′R)-praeruptorin A (8B), (3′S,4′S)-khellactone (9A), and (3′R,4′R)-khellactone (9B), respectively. Thereof, compounds 1A and 1B were new compounds, while compound 2A represents a new configuration for a known planar structure. Compounds 2A and 2B were isolated for the first time from Peucedani Radix. Above all, chiral LC-MS-guided isolation strategy is advantageous at rapid capturing new compounds from herbal medicines, providing an effective means for the separation of novel structures, especially new enantiomers.

【Objective】 To explore the effects of breastfeeding on the immune response of CD4⁺T lymphocytes in infants in non-inflammatory state, and to analyze the immunomodulatory significance of the whole composition of breast milk. 【Methods】 A retrospective cohort study was conducted. From January to September 2022, six-month-old infants who took physical examination in the Child Healthcare Department of Changzhou Children′s Hospital Affiliated to Nantong University, were selected based on inclusion criteria, and were divided into breastfeeding group (n=33) and formula feeding group (n=27) based on their feeding patterns. Flow cytometry was used to detect the percentage of CD4⁺ T cells, including helper T cell (Th) 1, Th2, Th17, regulatory T cell (Treg), and the levels of related cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, IL-17 in peripheral blood. The differences in these indicators between the two groups were compared. 【Results】 Compared with the formula feeding group, the breastfeeding group showed significantly higher percentages of Th1(t=3.038), Treg (t=2.088). The ratio of Th1 to Th2(Z=2.756), IL-10(Z=2.297) and IFN-γ (Z=2.076) in the peripheral blood of the breastfeeding group were also significantly higher. Conversely, the breastfeeding group had significantly lower percentage of Th17(Z=2.704) and IL-17A (t=2.187) (P<0.05). There was no significant difference the percentage of Th2, as well as in the levels of IL-2, IL-4, IL-6 and TNF-α between the two groups (P>0.05). 【Conclusions】 Breastfeeding has a regulatory effect on the immune response of infant CD4⁺ T lymphocytes. It promotes the development of Th1/Th2 towards Th1 and the immunomodulatory effect of Treg. Moreover, it inhibits the Th17 type immune response. These findings suggest that the complete composition of breast milk contributes to the development and maturation of infant immune system, enhancing immune defense and immune tolerance.

Objective To assess the prognostic significance of serum cystatin C(CysC)in combination with bedside renal ultrasound for patients diagnosed with sepsis-induced acute kidney injury(AKI).Methods The study cohort comprised 134 patients with sepsis-induced AKI who were admitted to our hospital between October 2019 and October 2023.Based on the 30 day prognosis,the patients were categorized into a survival group(n=93)and a death group(n=41).Collected clinical data included gender,age,heart rate,underlying diseases,treat-ment modalities,duration of hospital stay,basic biochemical indicators,and Acute Physiology and Chronic Health Evaluation Ⅱ(APACHEⅡ)scores.Renal function markers such as serum creatinine(SCr),blood urea nitrogen(BUN),and cystatin C levels were measured;renal blood flow resistance index(RI)was assessed using bedside routine ultrasound.Cox regression analysis was employed to evaluate factors influencing poor prognosis in patients with sepsis-induced AKI while analyzing the prognostic assessment value of combining cystatin C with bedside renal ultrasound through ROC curves.Results The death group exhibited significantly higher APACHEⅡ scores,SCr,BUN,CysC levels,and RI(all P<0.05)compared to the survival group.Cox regression analysis revealed that both CysC levels and RI were significant prognostic indicators(P<0.05).ROC curve analysis demonstrated that the combined assessment of CysC levels and RI yielded a high diagnostic accuracy of 97.5%in predicting outcomes for patients with sepsis-induced AKI.Conclusion CysC levels and bedside renal ultrasound can serve as prognostic indicators for patients with sepsis-induced acute kidney injury(AKI),thereby guiding clinical treatment.

Objective To investigate the impact of succinate dehydrogenase(SDH)on the modulation of human sperm functions.Methods The isolated human sperm were co-incubated with different concentrations(10,20,40 mmol/L)of SDH inhibitor disodium malonate for one or two hours.The activity of the SDH was measured by commercially available reagent kit,while the protein level of the SDH catalytic subunit SDHA was determined through Western blot analysis.Sperm functions were analyzed:1)The impact of disodium malonate on important mo-tility parameters of un-capcitated sperm including progressive motility rate(PR),total motility(TM),average pathvelocity(VAP)and the ability of capacitated sperm to penetrate viscous media were be assessed using a com-puter aided semen analysis system.2)Effect of disodium malonate on sperm survival rate was evaluated using the Eosin-Nigrosin microscopy.3)The incidence of acrosome reaction in capacitated sperm was be detected by PSA-FITC staining assay following disodium malonate treatment.Results Disodium malonate had no effect on expression of SDH catalytic subunit SHDA protein in human sperm.However,it inhibited the catalytic activity of the SDH,sperm forward motility,total motility,and the ability of sperm to penetrate viscous media.These inhibitory effects were positively correlated with the concentration of disodium malonate.Furthermore,disodium malonate had no any influence on the occurrence of spontaneous acrosome reaction in capacitated sperm.Conclusions Disodium mal-onate impairs human sperm motility by inhibiting succinate dehydrogenase activity.

Objective To explore the targets and pathways of Cynanchum wilfordii for treatment of ulcerative colitis(UC).Methods UPLC-QE-MS was used to identify the components of Cynanchum wilfordii ethanol extract,and their targets were screened using public databases for construction of the core protein-protein interaction(PPI)network and GO and KEGG enrichment analyses.Forty male C57 mice were randomized into normal control group,model group,mesalazine group and Cynanchum wilfordii group(n=10),and in the latter 3 groups,mouse UC models were established by treatment with 2.5%DSS and the latter 2 groups drug interventions by gavage.The therapeutic effect was evaluated by recording body weight changes and DAI score.Pathological changes of the colon tissue were observed with HE and AB-PAS staining,and JAK2 and STAT3 protein expressions were detected with Western blotting.The metabolites and metabolic pathways were identified by metabonomics analysis.Results We identified 240 chemical components in Cynanchum wilfordii alcoholic extracts,including 19 steroids.A total of 177 Cynanchum wilfordii targets,5406 UC genes,and 117 intersection genes were obtained.JAK2 and STAT3 were the core targets and significantly enriched in lipid and atherosclerosis pathways.Cynanchum wilfordii treatment significantly increased the body weight and decreased DAI score of UC mice(P<0.05),alleviated intestinal pathologies,and decreased JAK2 and STAT3 protein expressions in the colon tissues.Most of the 83 intersecting differential metabolites between the control,model and Cynanchum wilfordii groups were identified as glycerophospholipids,arachidonic acid,and amino acids involving glycerophospholipid metabolism and other pathways.Correlation analysis suggested that the core targets of Cynanchum wilfordii for UC participated in regulation of the metabolites.Conclusion Cynanchum wilfordii alleviates lipid and amino acid metabolism disorders to lessen UC in mice by regulating the core targets including JAK2 and STAT3 and the levels of endogenous metabolites.

Objective To explore the targets and pathways of Cynanchum wilfordii for treatment of ulcerative colitis(UC).Methods UPLC-QE-MS was used to identify the components of Cynanchum wilfordii ethanol extract,and their targets were screened using public databases for construction of the core protein-protein interaction(PPI)network and GO and KEGG enrichment analyses.Forty male C57 mice were randomized into normal control group,model group,mesalazine group and Cynanchum wilfordii group(n=10),and in the latter 3 groups,mouse UC models were established by treatment with 2.5%DSS and the latter 2 groups drug interventions by gavage.The therapeutic effect was evaluated by recording body weight changes and DAI score.Pathological changes of the colon tissue were observed with HE and AB-PAS staining,and JAK2 and STAT3 protein expressions were detected with Western blotting.The metabolites and metabolic pathways were identified by metabonomics analysis.Results We identified 240 chemical components in Cynanchum wilfordii alcoholic extracts,including 19 steroids.A total of 177 Cynanchum wilfordii targets,5406 UC genes,and 117 intersection genes were obtained.JAK2 and STAT3 were the core targets and significantly enriched in lipid and atherosclerosis pathways.Cynanchum wilfordii treatment significantly increased the body weight and decreased DAI score of UC mice(P<0.05),alleviated intestinal pathologies,and decreased JAK2 and STAT3 protein expressions in the colon tissues.Most of the 83 intersecting differential metabolites between the control,model and Cynanchum wilfordii groups were identified as glycerophospholipids,arachidonic acid,and amino acids involving glycerophospholipid metabolism and other pathways.Correlation analysis suggested that the core targets of Cynanchum wilfordii for UC participated in regulation of the metabolites.Conclusion Cynanchum wilfordii alleviates lipid and amino acid metabolism disorders to lessen UC in mice by regulating the core targets including JAK2 and STAT3 and the levels of endogenous metabolites.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.

Objective To investigate the anticoagulation status of patients with atrial fibrillation(AF)-related acute ischemic stroke(AIS)after revascularization therapy in the real world.Methods A retrospective study was performed on patients diagnosed as AIS and AF from Jan.2019 to Jan.2022 at The Second Affiliated Hospital of Nanchang University.Patients treated with intravenous thrombolysis(IVT),endovascular thrombectomy(EVT),or both were enrolled.Clinical information,timing of anticoagulation initiation,treatment regimens,and outcomes were documented and statistically analyzed.Additionally,a questionnaire was administered to the primary physicians to understand reasons for delaying or not initiating anticoagulation.Results A total of 189 patients with AF-related AIS met the screening criteria,including 86(45.5%)cases in the IVT group,63(33.3%)cases in the EVT group,and 40(21.2%)cases in the IVT+EVT group.The mean age of 189 patients was(72.90±9.23)years old.There were 93(49.2%)female patients.Anticoagulation was initiated within 14 d after revascularization therapy in 36.0%(68/189)of patients,with the highest rate in the IVT group(58.8%,40/68),followed by the EVT group(22.1%,15/68)and IVT+EVT group(19.1%,13/68).A significant difference was found in the proportion of patients receiving anticoagulation within 14 d among the 3 groups(P=0.020).Univariate analysis was performed on the clinical data of patients who initiated anticoagulation within 14 d after revascularization therapy(68 cases)and those who delayed or did not initiate anticoagulation(121 cases).The results showed that there were significant differences in the stroke history,National Institutes of Health stroke scale(NIHSS)score before revascularization therapy,Alberta Stroke Program early computed tomography score,modified Rankin scale(mRs)score before revascularization therapy,imaging characteristics(lesions near cortex,large infarction,severe stenosis or occlusion of major intracranial arteries),revascularization therapy method,NIHSS score 3 d after revascularization therapy,and intracranial hemorrhagic transformation after revascularization therapy between the 2 groups(all P＜0.05).Multivariate logistic regression analysis indicated that higher NIHSS scores 3 d after revascularization therapy(odds ratio[OR]=1.113,95%confidence interval[CI]1.053-1.176,P＜0.001)and the presence of intracranial hemorrhage after revascularization therapy(OR=6.098,95%CI 2.004-18.193,P=0.001)were significant factors that contraindicated the initiation of anticoagulation.Large infarcts(40.8%),infarct location(35.8%),and hemorrhagic transformation after stroke(40.8%)were the common reasons cited by physicians for not initiating anticoagulation.In the 90-d prognosis of patients with AF-related AIS,6 patients had bleeding events,and 116 patients had a good prognosis(mRS score of 0-2).The 90-d good prognosis rate in the initiated anticoagulation group within 14 d after revascularization therapy(89.7%,61/68)was significantly higher than that in the delayed or non-anticoagulation group(45.5%,55/121;P＜0.001).Conclusion For patients with AF-related AIS who receive IVT,EVT or IVT+EVT,it is safe to initiate anticoagulation early after revascularization therapy,but the timing of anticoagulation in most patients is later than the currently recommended anticoagulation timing.