ObjectiveTo observe the effects of Qingxin Jieyu granules （QXJYG） on FeCl₃-induced carotid artery thrombosis in rats and on the expression of thrombosis-related proteins tissue factor （TF） and tissue factor pathway inhibitor （TFPI） as well as the protein kinase B （Akt）/nuclear factor-κB （NF-κB） signaling pathway in EA.hy926 cells exposed to tumor necrosis factor-α （TNF-α）， thus preliminarily exploring the mechanism of QXJYG in inhibiting thrombosis. MethodsThirty-six SD rats were randomized into normal control， model， positive control （aspirin， 9 mg·kg^-1）， and low-， medium-， and high-dose （0.99， 1.98， 3.96 g·kg^-1， respectively） QXJYG groups （n=6）. The rats in the drug treatment groups were administrated with corresponding drugs， and those in the normal control group and model group were given an equal volume of distilled water. After 14 consecutive days of prophylactic gavage， the rat model of common carotid artery thrombosis was established with 45% FeCl₃ solution， and the blood vessels were collected and the wet weight of thrombus was weighed by an electronic balance （precision of 1/10 000）. The thrombosis in the common carotid artery of each group of rats was observed by hematoxylin-eosin staining. The plasma levels of von Willebrand factor （vWF）， platelet endothelial cell adhesion molecule-1 （PECAM-1）， tissue-type plasminogen activator （t-PA）， and plasminogen activator inhibitor-1 （PAI-1） were determined by enzyme-linked immunosorbent assay. An endothelial cell injury model was established by treating EA.hy926 human umbilical vein endothelial cells with TNF-α. The cell counting kit-8 method was used to screen the intervention concentrations of QXJYG. Western blot was employed to determine the protein levels of TF， TFPI， Akt， p-Akt， NF-κB p65， and p-NF-κB p65 in each group of cells. ResultsThe animal experiment showed that compared with the normal control group， the model group showed an increase in carotid artery thrombus weight （P<0.05）， with unclear vascular structure and extensive thrombosis in the lumen. In addition， the plasma levels of vWF， PECAM-1， and PAI-1 were elevated， while the t-PA level became lowered （P<0.05） in the model group. Compared with the model group， the aspirin and QXJYG groups showed reductions in the weight of FeCl₃-induced carotid artery thrombi （P<0.05） and thrombosis in the lumen， declines in plasma levels of PECAM-1 and PAI-1， and an elevation in the t-PA level （P<0.05）. Moreover， the QXJYG groups showed reductions in the plasma level of vWF （P<0.05）， which， however， had no significant difference between the aspirin group and the model group. The cell experiments indicated that 31.25， 62.5， 125， 250， 500 mg·L^-1 QXJYG had no effect on the viability of EA.hy926 cells. Therefore， 250， 500 mg·L^-1 QXJYG were selected as the intervention concentrations for subsequent experiments. Western blotting results showed that compared with the control group， the TNF-α stimulation downregulated the expression of TFPI （P<0.05）， upregulated the expression of TF， and increased the ratios of p-Akt/Akt and p-NF-κB p65/NF-κB p65 （P<0.05） in EA.hy926 cells. Compared with the model group， the intervention with QXJYG upregulated the expression of TFPI （P<0.05）， inhibited the expression of TF， and decreased the ratios of p-Akt/Akt and p-NF-κB p65/NF-κB p65 （P<0.05）. ConclusionQXJYG has the effect of inhibiting thrombosis and regulating the expression of TF and TFPI in endothelial cells exposed to TNF-α by suppressing the abnormal activation of the Akt/NF-κB signaling pathway.

Objective:The transcriptome data was utilized to screen cuproptosis-related genes(CRGs)in oral squamous cell car-cinoma(OSCC),and the characteristic genes were identified for constructing a prognostic model for predicting patients'survival time.Methods:OSCC transcriptome gene expression and clinical data were obtained from TCGA and GEO.Through Lasso regres-sion analysis and Cox regression analysis,relevant prognostic genes were screened and prognostic models were constructed.Ac-cording to the median value of risk scores,patients were divided into high and low risk groups,and their survival rates were com-pared.Finally,the predictive performance of the model was verified.Results:In this study,9 characteristic genes with prognostic value(ENO2,P4HA1,SLC2A3,AQP1,PLS1,NXPH4,CTSG,TRAC,THBS1)were screened out and a 9-gene prognostic model was constructed.The survival rate of high-risk group based on prognostic model was significantly lower than that of low-risk group.The area under curve(AUC)of receiver operating characteristic curve(ROC)was 0.701,0.729 and 0.702 at 1 year,3 years and 5 years,respectively,which verifies that the risk model has good predictive performance.The nomogram predicted that the 1-year,3-year,and 5-year survival probabilities of OSCC patients are 89.6％,72.4％,and 63.9％respectively,and the cal-ibration curve confirmed the accuracy of the nomogram prediction.Conclusion:The 9-gene prognostic model based on CRGs screening could predict the prognosis of OSCC patients,which is helpful for clinical personalized treatment of OSCC patients and prediction of their survival rate.

Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

Objective:To explore the role of Caveolin-1 (CAV-1) in radiation-induced premature senescence of vascular endothelial cells.Methods:A cell model with stable knockdown of CAV-1 was constructed in human microvascular endothelial cells (HMEC-1) by lentiviral transfection using puromycin screening. The cells were divided into NC group and sh-CAV-1 group based on whether they were infected with lentivirus shRNA-CAV-1. The protein expression levels of CAV-1, p53 and p21 were detected by Western blot at 24, 48, and 72 h after 0, 2, and 4 Gy X-ray irradiation. The β-galactosidase staining kit was used to detect β-galactosidase in cells. CCK-8 kit was used to detect cell viability, and vascular endothelial cell function was detected by vascular tube-forming assay.Results:CAV-1 protein expression was significantly decreased at 48 h after 2 and 4 Gy X-ray irradiation ( t=3.50, 3.89, P < 0.05), and β-galactosidase in sh-CAV-1 group was significantly increased at 72 h after 0, 2 and 4 Gy X-ray irradiation ( t=12.91, 11.54, 6.04, P < 0.05) compared with the NC group. Knockdown of CAV-1 resulted in the decrease in the expression level of the cellular senescence-associated protein p53 protein ( t=4.09, 3.13, 3.43, P < 0.05), but increase in the expression level of p21 protein ( t=-3.63, -3.33, -3.06, P < 0.05). Compared with the NC group, knockdown CAV-1 significantly decreased cell viability ( t=2.97-25.89, P<0.05) and reduced vessel-forming capacity ( t=3.39-39.68, P < 0.05). Conclusions:CAV-1 is involved in the process of radiation-induced premature senescence of vascular endothelial cells through positive regulation of p53 and negative regulation of p21.

OBJECTIVE To analyze the reasons for the outbreak of suspected carbapenem-resistant Acinetobacter baumannii(CRAB)infections in the neurosurgical intensive care unit to take comprehensive prevention and con-trol measures to control the spread of infections,and to provide a reference for the control of multidrug-resistant organisms(MDRO)infections in intensive care units.METHODS Epidemiological data were collected from five patients with CRAB infections in the neurosurgical intensive care unit of Gansu Provincial People's Hospital in Mar.2024,and environmental hygiene monitoring and comprehensive infection prevention and control measures were conducted.RESULTS A total of five cases of lower respiratory tract infections occurred,all of which were hospital-acquired infections,and all patients had consistent spatial-temporal distribution.A.baumannii was detec-ted in the sputum cultures of all patients,and all were resistant to carbapenems,aminoglycosides and some β-lac-tam drugs.Before the implementation of comprehensive prevention and control measures,a total of 25 samples from the surfaces and the hands of health workers were collected,with a pass rate of 52.00％;CRAB was detected on the surface of treatment carts and the wall of sputum suction bottles in the treatment room,with a detection rate of 8.00％.After the implementation,a total of 24 samples were collected,with a pass rate of 100.00％,and the difference was statistically significant when compared with the pre-implementation period(χ2=16.987,P<0.001).Through the implementation of comprehensive measures such as isolation,standardization of the ward cleaning and disinfection process,strengthening hand hygiene,standardization of sterile operations and personnel management,the indicators such as hand hygiene compliance rate,the execution rate of multi-drug-resistant bac-terial preventive and control measures and the fluorescent marker removal rate were all improved,and the suspec-ted CRAB infection outbreak was timely controlled.CONCLUSION This suspected CRAB hospital-acquired infec-tion outbreak may be related to a variety of factors,and timely environmental hygiene monitoring and comprehen-sive infection prevention and control measures can effectively control the spread of CRAB infection.

Objective To observe the effect of Qing-Xin-Jie-Yu granule(QXJYG)on the formation of thrombosis in the rat model of arteriovenous bypass thrombosis and the adenosine diphosphate(ADP)-induced platelet aggregation.Methods Thirty-six male SD rats were randomly divided into normal control group,model group,clopidogrel positive control group,QXJYG low-dose,medium-dose and high-dose groups,with 6 rats in each group.The dose of clopidogrel positive control group was 6.74 mg/(kg?d),the dosages of QXJYG in low,medium and high groups were 0.99,1.98,3.96 g/(kg?d),respectively,normal control group and model group were given equal volume of distilled water,and continuous prophylactic intragastric administration for 14 days,once a day.One hour after the final administration,the rats were anesthetized,and the arteriovenous bypass thrombosis model was established by using a polyethylene tube as the arteriovenous bypass bridge(except control group).The thrombus was extracted after 15 min and its weight was weighed by 1/10,000th precision electronic balance.The levels of thromboxane B2(TXB2)and 6-keto-prostaglandin F1α(6-keto-PGF1α)in plasma were determined by ELISA kits.The rate of platelet aggregation induced by ADP in each group was measured using a microplate reader by turbidimetric method.Results Compared with the control group,the weight of arteriovenous bypass thrombus was significantly higher,the level of TXB2 in plasma was significantly higher,while the level of 6-keto-PGF1α was significantly lower,and platelet aggregation was significantly higher after ADP induction in the model group(P＜0.05).Compared with the model group,the weight of arteriovenous bypass thrombosis in clopidogrel positive control group and QXJYG dose groups was significantly decreased(P＜0.05);the inhibition rate of thrombosis formation was 53.80％,23.96％,33.63％,and 32.59％,respectively.The content of TXB2 in plasma was significantly decreased,the content of 6-keto-PGF1α was significantly increased;additionally,the platelet aggregation rate induced by ADP was reduced in clopidogrel positive control group and QXJYG group.Meanwhile,there was a dose-dependence between different doses in QXJYY group(P＜0.05),and the inhibition rate of platelet aggregation was 86.90％,26.17％,38.87％,54.48％,respectively.Conclusion QXJYG can prevent thrombosis formation in the rat model of arteriovenous bypass thrombosis and inhibit platelet aggregation induced by ADP.

Objective:To explore the role of Caveolin-1 (CAV-1) in radiation-induced premature senescence of vascular endothelial cells.Methods:A cell model with stable knockdown of CAV-1 was constructed in human microvascular endothelial cells (HMEC-1) by lentiviral transfection using puromycin screening. The cells were divided into NC group and sh-CAV-1 group based on whether they were infected with lentivirus shRNA-CAV-1. The protein expression levels of CAV-1, p53 and p21 were detected by Western blot at 24, 48, and 72 h after 0, 2, and 4 Gy X-ray irradiation. The β-galactosidase staining kit was used to detect β-galactosidase in cells. CCK-8 kit was used to detect cell viability, and vascular endothelial cell function was detected by vascular tube-forming assay.Results:CAV-1 protein expression was significantly decreased at 48 h after 2 and 4 Gy X-ray irradiation ( t=3.50, 3.89, P < 0.05), and β-galactosidase in sh-CAV-1 group was significantly increased at 72 h after 0, 2 and 4 Gy X-ray irradiation ( t=12.91, 11.54, 6.04, P < 0.05) compared with the NC group. Knockdown of CAV-1 resulted in the decrease in the expression level of the cellular senescence-associated protein p53 protein ( t=4.09, 3.13, 3.43, P < 0.05), but increase in the expression level of p21 protein ( t=-3.63, -3.33, -3.06, P < 0.05). Compared with the NC group, knockdown CAV-1 significantly decreased cell viability ( t=2.97-25.89, P<0.05) and reduced vessel-forming capacity ( t=3.39-39.68, P < 0.05). Conclusions:CAV-1 is involved in the process of radiation-induced premature senescence of vascular endothelial cells through positive regulation of p53 and negative regulation of p21.

Objective:The transcriptome data was utilized to screen cuproptosis-related genes(CRGs)in oral squamous cell car-cinoma(OSCC),and the characteristic genes were identified for constructing a prognostic model for predicting patients'survival time.Methods:OSCC transcriptome gene expression and clinical data were obtained from TCGA and GEO.Through Lasso regres-sion analysis and Cox regression analysis,relevant prognostic genes were screened and prognostic models were constructed.Ac-cording to the median value of risk scores,patients were divided into high and low risk groups,and their survival rates were com-pared.Finally,the predictive performance of the model was verified.Results:In this study,9 characteristic genes with prognostic value(ENO2,P4HA1,SLC2A3,AQP1,PLS1,NXPH4,CTSG,TRAC,THBS1)were screened out and a 9-gene prognostic model was constructed.The survival rate of high-risk group based on prognostic model was significantly lower than that of low-risk group.The area under curve(AUC)of receiver operating characteristic curve(ROC)was 0.701,0.729 and 0.702 at 1 year,3 years and 5 years,respectively,which verifies that the risk model has good predictive performance.The nomogram predicted that the 1-year,3-year,and 5-year survival probabilities of OSCC patients are 89.6％,72.4％,and 63.9％respectively,and the cal-ibration curve confirmed the accuracy of the nomogram prediction.Conclusion:The 9-gene prognostic model based on CRGs screening could predict the prognosis of OSCC patients,which is helpful for clinical personalized treatment of OSCC patients and prediction of their survival rate.

OBJECTIVE To explore the risk factors for hospital-associated infections(HAI)in dead patients so as to provide bases for development of prevention strategies for the hospital-associated infections.METHODS Totally 560 patients who died due to non-SARS-CoV-2 infections and hospitalized in Gansu Provincial People's Hospital for more than 48 hours in 2019 and 2023 were retrospectively analyzed,70 of whom had HAI and were assigned as the HAI group,and the rest of 490 patients were assigned as the non-HAI group.The incidence of HAI,major causes of death,risk factors and economic burden were observed.RESULTS The incidence of HAI was 12.50％a-mong the dead patients,the lower respiratory tract infection was the predominant type of infection,and the re-spiratory failure was the primary and direct cause of death in the HAI group;the total treatment cost of the HAI group was higher than that of the non-HAI group(P＜0.05).Univariate analysis showed that there were signifi-cant differences in the length of hospital stay,multidrug-resistant organisms infections,combined use of antibiot-ics,surgery,hemodialysis/peritoneal dialysis,invasive procedures(drainage,puncture,intubation,tracheoto-my),use of equipment like ventilator or urinary catheter,central venous catheter indwelling,blood transfusion,and use of immunosuppressors/glucocorticoids between the HAI group and the non-HAI group(P＜0.05),while there were no significant differences in the sex,age,underlying diseases,modified early warning score(MEWS)and major diagnosis and chemoradiotherapy between the two groups.Multivariate analysis indicated that the length of hospital stay more than 14 days,history of surgery and combined use of antibiotics were the risk factors for HAI in the dead patients(P＜0.05).CONCLUSION It is necessary for the hospital to identify the high-risk pa-tients as early as possible,pay close attention to the lower respiratory tract infection,reduce the risk of HAI by shortening the length of hospital stay,standardizing the invasive procedures and reasonably using antibiotics so as to improve the prognosis of the patients.

Objective To observe the effect of Qing-Xin-Jie-Yu granule(QXJYG)on the formation of thrombosis in the rat model of arteriovenous bypass thrombosis and the adenosine diphosphate(ADP)-induced platelet aggregation.Methods Thirty-six male SD rats were randomly divided into normal control group,model group,clopidogrel positive control group,QXJYG low-dose,medium-dose and high-dose groups,with 6 rats in each group.The dose of clopidogrel positive control group was 6.74 mg/(kg?d),the dosages of QXJYG in low,medium and high groups were 0.99,1.98,3.96 g/(kg?d),respectively,normal control group and model group were given equal volume of distilled water,and continuous prophylactic intragastric administration for 14 days,once a day.One hour after the final administration,the rats were anesthetized,and the arteriovenous bypass thrombosis model was established by using a polyethylene tube as the arteriovenous bypass bridge(except control group).The thrombus was extracted after 15 min and its weight was weighed by 1/10,000th precision electronic balance.The levels of thromboxane B2(TXB2)and 6-keto-prostaglandin F1α(6-keto-PGF1α)in plasma were determined by ELISA kits.The rate of platelet aggregation induced by ADP in each group was measured using a microplate reader by turbidimetric method.Results Compared with the control group,the weight of arteriovenous bypass thrombus was significantly higher,the level of TXB2 in plasma was significantly higher,while the level of 6-keto-PGF1α was significantly lower,and platelet aggregation was significantly higher after ADP induction in the model group(P＜0.05).Compared with the model group,the weight of arteriovenous bypass thrombosis in clopidogrel positive control group and QXJYG dose groups was significantly decreased(P＜0.05);the inhibition rate of thrombosis formation was 53.80％,23.96％,33.63％,and 32.59％,respectively.The content of TXB2 in plasma was significantly decreased,the content of 6-keto-PGF1α was significantly increased;additionally,the platelet aggregation rate induced by ADP was reduced in clopidogrel positive control group and QXJYG group.Meanwhile,there was a dose-dependence between different doses in QXJYY group(P＜0.05),and the inhibition rate of platelet aggregation was 86.90％,26.17％,38.87％,54.48％,respectively.Conclusion QXJYG can prevent thrombosis formation in the rat model of arteriovenous bypass thrombosis and inhibit platelet aggregation induced by ADP.