AIM:To investigate the protective effects of Klotho protein against hypoxia/reoxygenation(H/R)-in-duced damage in rat cardiomyocytes,and to elucidate its underlying mechanisms.METHODS:Rat cardiomyocyte H9c2 cells were divided into 4 groups:control,H/R,low-concentration(1 μmol/L)Klotho+H/R,and high-concentration(10 μmol/L)Klotho+H/R groups.Cells were pretreated with Klotho at specified concentrations before induction of H/R injury.Flow cytometry was used to determine cardiomyocyte apoptosis rates,while reactive oxygen species(ROS)levels were measured using the DCFH-DA probe.Additionally,superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were assessed using biochemical assay kits.Mitochondrial membrane potential was evaluated using the JC-1 as-say,and activity of caspase-3 and caspase-9 was quantified.Western blot analysis was conducted to measure the protein expression of cytochrome C(Cyt-C),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in cardio-myocytes from each group.RESULTS:Compared with the control group,the H/R group exhibited significantly increased apoptosis rates(P<0.05),elevated ROS levels and MDA content,decreased SOD activity(P<0.05),reduced mitochon-drial membrane potential(P<0.05),increased caspase-3 and caspase-9 activity(P<0.05),decreased mitochondrial Cyt-C and Bcl-2 protein expression(P<0.05),and increased cytoplasmic Cyt-C and Bax protein expression(P<0.05).In comparison with the H/R group,both low-and high-concentration Klotho treatments significantly reduced cardiomyocyte apoptosis(P<0.05),lowered ROS levels and MDA content(P<0.05),increased SOD activity(P<0.05),restored mito-chondrial membrane potential(P<0.05),decreased caspase-3 and caspase-9 activity(P<0.05),increased mitochondrial Cyt-C and Bcl-2 expression(P<0.05),and decreased cytoplasmic Cyt-C and Bax expression(P<0.05).Notably,the high-concentration Klotho group demonstrated more pronounced protective effects(P<0.05).CONCLUSION:Klotho protein exerts protective effects against H/R-induced cardiomyocyte injury,possibly by inhibiting mitochondria-mediated apoptosis.

Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

Objective To investigate the impact of pharmaceutical services provided by clinical pharmacists on rational drug use and cost control in hepatobiliary surgery under the Diagnosis Related Groups(DRGs)payment model,aming to provide evidence for improving the rationality of drug therapy and saving medical costs.Methods Patients classified under DRGs disease codes HB15,HB13,and HB11 from November 2022 to April 2024 were selected as study subjects.A total of 195 patients were included,with 106 in the intervention group and 89 in the control group.The intervention group received multidimensional clinical pharmaceutical services in addition to the standard care provided to the control group.The rational drug use rate,medication costs,total hospitalization expenses,and length of hospital stay were observed between the two groups.A cost-benefit analysis was employed to evaluate the economic impact of providing pharmaceutical services to hepatobiliary surgical patients.The cost indicator was the clinical pharmacy services cost,and the benefit indicators were the reductions in total hospitalization expenses and medication costs.The benefit-cost ratio(B/C)was calculated,and sensitivity analysis was performed.Results The intervention group showed significantly higher rational use rates of prophylactic antimicrobial agents(drug selection:83.96％vs.46.07％,P＜0.01;treatment duration:84.91％vs.56.18％,P＜0.01)and parenteral nutrition drugs(97.17％vs.73.03％,P＜0.01)compared to the control group.Additionally,the intervention group had significantly reduced the length of hospital stay,total hospitalization expenses,medication costs,and insurance over-expenditure compared to the control group(P＜0.05).Furthermore,clinical pharmacist intervention led to a reduction in medication costs by 4 320.05(2 555.00,5 088.25)yuan(CNY)and total hospitalization expenses by 8 891.12(5 135.05,10 074.03)yuan(CNY).The B/C ratios were 14.24(8.42,16.77)and 29.30(16.92,33.20),respectively,indicating economic efficiency.Sensitivity analysis supported these results.Conclusion Under the DRGs payment model,clinical pharmaceutical services guided by drug therapy pathways contribute to improving rational drug use in hepatobiliary surgery and provide clear economic benefits,playing a positive role in reducing medical costs.

Objective:To evaluate the value of ultrasound radiomics combined with machine learning for early diagnosis of seronegative Hashimoto’s thyroiditis (SN-HT) .Methods:This retrospective study included 164 patients from Liaoning Provincial People’s Hospital , Lixin County People’s Hospital, Linghai Dalinghe Hospital, Fengcheng Phoenix Hospital, who underwent thyroidectomy for solitary nodules with normal thyroid function between Nov. 2016 and Jan. 2024. Postoperative pathology confirmed Hashimoto’s thyroiditis (HT) in some cases, who were further categorized into antibody-positive and antibody-negative groups based on serum antibody status. Patients without Hashimoto’s thyroiditis served as the control group. A total of 298 ultrasound images were analyzed. Radiomics features were extracted from hypoechoic non-nodular areas within 0.5 cm surrounding the tumor. Two senior pathologists and two senior ultrasound physicians independently assessed lymphocytic infiltration, eosinophilic changes of follicular epithelium, and the proportion of hypoechoic areas in pathology and ultrasound images, respectively. A machine learning model, CCH-NET, was developed using linear regression and t-distributed stochastic neighbor embedding (t-SNE) techniques. The dataset was divided into a training set (80%) and a validation set (20%) to compare the diagnostic accuracy of CCH-NET with that of senior ultrasound physicians. Results:In internal validation, CCH-NET achieved a diagnostic accuracy of 88.89% for both antibody-positive and antibody-negative groups, significantly higher than the 66.67% accuracy of senior ultrasound physicians ( P<0.01). In external validation, CCH-NET achieved 75.00% and 66.67% accuracy for the two groups, compared to 50.00% by senior ultrasound physicians. For the control group, both methods achieved 93.33% accuracy. The AUC of CCH-NET was 0.848, outperforming senior ultrasound physicians (0.681) ,demonstrating superior diagnostic performance. Conclusion:The radiomics-based CCH-NET model, using non-nodular hypoechoic areas as a specific indicator, can accurately identify early SN-HT in euthyroid patients. It significantly outperforms senior ultrasound physicians, improving diagnostic accuracy and reducing missed diagnoses.

Objective:To evaluate the value of ultrasound radiomics combined with machine learning for early diagnosis of seronegative Hashimoto’s thyroiditis (SN-HT) .Methods:This retrospective study included 164 patients from Liaoning Provincial People’s Hospital , Lixin County People’s Hospital, Linghai Dalinghe Hospital, Fengcheng Phoenix Hospital, who underwent thyroidectomy for solitary nodules with normal thyroid function between Nov. 2016 and Jan. 2024. Postoperative pathology confirmed Hashimoto’s thyroiditis (HT) in some cases, who were further categorized into antibody-positive and antibody-negative groups based on serum antibody status. Patients without Hashimoto’s thyroiditis served as the control group. A total of 298 ultrasound images were analyzed. Radiomics features were extracted from hypoechoic non-nodular areas within 0.5 cm surrounding the tumor. Two senior pathologists and two senior ultrasound physicians independently assessed lymphocytic infiltration, eosinophilic changes of follicular epithelium, and the proportion of hypoechoic areas in pathology and ultrasound images, respectively. A machine learning model, CCH-NET, was developed using linear regression and t-distributed stochastic neighbor embedding (t-SNE) techniques. The dataset was divided into a training set (80%) and a validation set (20%) to compare the diagnostic accuracy of CCH-NET with that of senior ultrasound physicians. Results:In internal validation, CCH-NET achieved a diagnostic accuracy of 88.89% for both antibody-positive and antibody-negative groups, significantly higher than the 66.67% accuracy of senior ultrasound physicians ( P<0.01). In external validation, CCH-NET achieved 75.00% and 66.67% accuracy for the two groups, compared to 50.00% by senior ultrasound physicians. For the control group, both methods achieved 93.33% accuracy. The AUC of CCH-NET was 0.848, outperforming senior ultrasound physicians (0.681) ,demonstrating superior diagnostic performance. Conclusion:The radiomics-based CCH-NET model, using non-nodular hypoechoic areas as a specific indicator, can accurately identify early SN-HT in euthyroid patients. It significantly outperforms senior ultrasound physicians, improving diagnostic accuracy and reducing missed diagnoses.

AIM:To investigate the protective effects of Klotho protein against hypoxia/reoxygenation(H/R)-in-duced damage in rat cardiomyocytes,and to elucidate its underlying mechanisms.METHODS:Rat cardiomyocyte H9c2 cells were divided into 4 groups:control,H/R,low-concentration(1 μmol/L)Klotho+H/R,and high-concentration(10 μmol/L)Klotho+H/R groups.Cells were pretreated with Klotho at specified concentrations before induction of H/R injury.Flow cytometry was used to determine cardiomyocyte apoptosis rates,while reactive oxygen species(ROS)levels were measured using the DCFH-DA probe.Additionally,superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were assessed using biochemical assay kits.Mitochondrial membrane potential was evaluated using the JC-1 as-say,and activity of caspase-3 and caspase-9 was quantified.Western blot analysis was conducted to measure the protein expression of cytochrome C(Cyt-C),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in cardio-myocytes from each group.RESULTS:Compared with the control group,the H/R group exhibited significantly increased apoptosis rates(P<0.05),elevated ROS levels and MDA content,decreased SOD activity(P<0.05),reduced mitochon-drial membrane potential(P<0.05),increased caspase-3 and caspase-9 activity(P<0.05),decreased mitochondrial Cyt-C and Bcl-2 protein expression(P<0.05),and increased cytoplasmic Cyt-C and Bax protein expression(P<0.05).In comparison with the H/R group,both low-and high-concentration Klotho treatments significantly reduced cardiomyocyte apoptosis(P<0.05),lowered ROS levels and MDA content(P<0.05),increased SOD activity(P<0.05),restored mito-chondrial membrane potential(P<0.05),decreased caspase-3 and caspase-9 activity(P<0.05),increased mitochondrial Cyt-C and Bcl-2 expression(P<0.05),and decreased cytoplasmic Cyt-C and Bax expression(P<0.05).Notably,the high-concentration Klotho group demonstrated more pronounced protective effects(P<0.05).CONCLUSION:Klotho protein exerts protective effects against H/R-induced cardiomyocyte injury,possibly by inhibiting mitochondria-mediated apoptosis.

Objective To investigate the impact of pharmaceutical services provided by clinical pharmacists on rational drug use and cost control in hepatobiliary surgery under the Diagnosis Related Groups(DRGs)payment model,aming to provide evidence for improving the rationality of drug therapy and saving medical costs.Methods Patients classified under DRGs disease codes HB15,HB13,and HB11 from November 2022 to April 2024 were selected as study subjects.A total of 195 patients were included,with 106 in the intervention group and 89 in the control group.The intervention group received multidimensional clinical pharmaceutical services in addition to the standard care provided to the control group.The rational drug use rate,medication costs,total hospitalization expenses,and length of hospital stay were observed between the two groups.A cost-benefit analysis was employed to evaluate the economic impact of providing pharmaceutical services to hepatobiliary surgical patients.The cost indicator was the clinical pharmacy services cost,and the benefit indicators were the reductions in total hospitalization expenses and medication costs.The benefit-cost ratio(B/C)was calculated,and sensitivity analysis was performed.Results The intervention group showed significantly higher rational use rates of prophylactic antimicrobial agents(drug selection:83.96％vs.46.07％,P＜0.01;treatment duration:84.91％vs.56.18％,P＜0.01)and parenteral nutrition drugs(97.17％vs.73.03％,P＜0.01)compared to the control group.Additionally,the intervention group had significantly reduced the length of hospital stay,total hospitalization expenses,medication costs,and insurance over-expenditure compared to the control group(P＜0.05).Furthermore,clinical pharmacist intervention led to a reduction in medication costs by 4 320.05(2 555.00,5 088.25)yuan(CNY)and total hospitalization expenses by 8 891.12(5 135.05,10 074.03)yuan(CNY).The B/C ratios were 14.24(8.42,16.77)and 29.30(16.92,33.20),respectively,indicating economic efficiency.Sensitivity analysis supported these results.Conclusion Under the DRGs payment model,clinical pharmaceutical services guided by drug therapy pathways contribute to improving rational drug use in hepatobiliary surgery and provide clear economic benefits,playing a positive role in reducing medical costs.

Consecutively hospitalized patients with confirmed coronavirus disease 2019 (COVID-19) in Wuhan, China were retrospectively enrolled from January 2020 to March 2020 to investigate the association between the use of renin-angiotensin system inhibitor (RAS-I) and the outcome of this disease. Associations between the use of RAS-I (angiotensin-converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB)), ACEI, and ARB and in-hospital mortality were analyzed using multivariate Cox proportional hazards regression models in overall and subgroup of hypertension status. A total of 2771 patients with COVID-19 were included, with moderate and severe cases accounting for 45.0% and 36.5%, respectively. A total of 195 (7.0%) patients died. RAS-I (hazard ratio (HR)= 0.499, 95% confidence interval (CI) 0.325-0.767) and ARB (HR = 0.410, 95% CI 0.240-0.700) use was associated with a reduced risk of all-cause mortality among patients with COVID-19. For patients with hypertension, RAS-I and ARB applications were also associated with a reduced risk of mortality with HR of 0.352 (95% CI 0.162-0.764) and 0.279 (95% CI 0.115-0.677), respectively. RAS-I exhibited protective effects on the survival outcome of COVID-19. ARB use was associated with a reduced risk of all-cause mortality among patients with COVID-19.
Angiotensin Receptor Antagonists/therapeutic use* ; Angiotensin-Converting Enzyme Inhibitors/therapeutic use* ; COVID-19 ; Humans ; Hypertension/drug therapy* ; Renin-Angiotensin System ; Retrospective Studies

Objective:To investigate the prevalence and risk factors of diabetic retinopathy (DR) in patients in Tibet.Methods:A total of 239 patients with DR who received treatment in Department of Endocrinology and Metabolism, Hospital of Chengdu Office of People's Government of Tibet Autonomous Region from December 2017 to December 2018 were included in this study. They were divided into Han nationality and Zang nationality groups according to ethnicity. The condition of DR was evaluated with nonmydriatic ocular fundus photography according to the staging criteria of the severity of retinopathy.Results:The prevalence of DR in Tibet was 18.0％. The prevalence of DR in Tibetan and Han patients with diabetes was 17.5% and 19.2%, respectively. There was no significant difference in the prevalence of DR between Tibetan and Han patients with diabetes ( χ2 = 0.10, P = 0.754). Logistic regression analysis revealed that the risk factors of developing DR in Tibet included diabetes duration ( OR = 1.14, 95% CI: 1.05-1.24, P < 0.05), insulin therapy ( OR = 2.74, 95% CI: 1.09-6.89, P < 0.05), fasting plasma glucose ( OR = 1.37, 95% CI: 1.07-1.75, P < 0.05) and hypertension ( OR = 1.98, 95% CI: 1.02-3.86, P < 0.05). Diabetes duration and fasting plasma glucose are independent risk factors of DR. However, although elevated glycated hemoglobin levels were high in Tibet, they could not be used to predict the risk for developing DR ( OR = 1.01, 95% CI: 0.82-1.25, P > 0.05). Conclusion:Hyperglycemia is an important risk factor of developing DR in Tibet. However, elevated glycated hemoglobin levels cannot be used to predict the risk of developing DR in Tibet. Findings from this study fill the gap in the research on DR prevalence and ethic difference of DR prevalence, providing scientific evidence for prevention and treatment of DR in high-altitude areas.