Endovascular therapy (EVT) is the standard treatment for acute ischemic stroke (AIS) due to large vessel occlusion (LVO), which can effectively improve patient outcome by timely restoring cerebral blood flow perfusion. The benefits of EVT are time-dependent and significant when initiated within 6 hours of onset. The onset-to-puncture time (OPT) is divided into onset-to-door time (ODT) and door-to-puncture time (DPT). ODT is the time from symptom onset to arrival at the emergency department, reflecting the efficiency of prehospital emergency treatment; DPT is the time from emergency admission to completion of femoral artery puncture, reflecting the efficiency of in-hospital diagnosis and treatment. Despite various efforts were made in the early stage to enable 41.6% of patients with AIS who received EVT achieved a door-to-device time (DDT) of <120 min, OPT is still not ideal, and issues such as inaccurate prehospital assessments and delayed in-hospital diagnosis and treatment remain severe. This article elaborates on the impact and possible mechanisms of OPT on the outcome of patients with AIS undergoing EVT, and discusses the influencing factors and optimization strategies of OPT.

Objective To observe the effects of auricular point sticking on chronic soft tissue injury of knee joint caused by military training.Methods A total of 77 patients with chronic soft tissue injury of the knee caused by military training admitted to No.901 Hospital of Joint Logistics Support Force from January to November 2022 were enrolled in this study.Auricular point sticking therapy was applied in all the patients.Visual analogue scale(VAS)score,the range of motion of the knees and knee temperature were recorded before and after treatment.Results The VAS score was 6.10±1.23,3.03±0.95,and 2.70±0.93 before treatment,15 min and 24 h after treatment,respectively.The VAS score after treatment were significantly lower than that before treatment(P<0.01).The range of motion of the knees was(79.08±11.59)°,(104.79±8.49)°,and(108.08±9.36)° before treatment,15 min and 24 h after treatment,respectively.The knee flexion after treatment were significantly higher than that before treatment(P<0.01).The knee temperature at 15 min after treatment was significantly higher than that before treatment([30.68±1.38]℃vs.[29.30±1.41]℃,P<0.01).Conclusion Auricular point sticking can reduce the pain of patients with chronic soft tissue injury of the knee and increase the range of joint motion.It is safe and simple and has good compliance.

Objective:To explore the effect of Chaibei Zhixian Decoction on intestinal flora and T helper 17/regulatory T cell(Th17/Treg)immune balance in epileptic rats by regulating mammalian target of rapamycin/hypoxia-inducible factor-1α(mTOR/HIF-1α)pathway.Methods:Sixty rats were randomly divided into Sham group,Model group,low-dose Chaibei Zhixian Decoction group(CBZXD-Low)and high-dose Chaibei Zhixian Decoction group(CBZXD-High),with 15 rats in each group.Lateral ventricular injec-tion 1 μl kainic acid(1.5 μg/μl)was used to prepare epileptic rat model,Sham group was injected with an equal amount of physiological saline.CBZXD-Low group and CBZXD-High group were given Chaibei Zhixian Decoction by gavage daily(8.48 g/kg,16.96 g/kg),while Sham group and model group were given an equal amount of physiological saline by gavage.After continuous gavage for 4 weeks,HE staining and Nissl staining were used to detect pathological damage in hippocampal tissue;RT-qPCR was used to detect Th17/Treg related marker mRNA levels in spleen tissue and mTOR,HIF-1α mRNA levels in hippocampus tissue;flow cytometry was used to de-tect Th17/Treg cell ratio in spleen tissue;16S rRNA sequencing of feces in colon segment was performed;Western blot was used to de-tect mTOR and HIF-1α protein levels in hippocampal tissue.Results:Compared with Sham group,Model group rats suffered from spontaneous epilepsy,the arrangement of hippocampal neurons was disordered and severely lacking,intracellular Nissl bodies was de-creased,RORγt,IL-17A mRNA levels in spleen tissue were increased,Foxp3,CTLA-4 and GITR mRNA levels were decreased,the proportion of Th17 cells and Th17/Treg were increased,the proportion of Treg cell was decreased,intestinal flora α/β diversity were decreased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were increased,the proportion of beneficial bacte-ria such as Bacteroides,Lactobacillus,Prevotella and Akkermansia were decreased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levelsin hippocampal tissue were increased(P<0.05);compared with Model group,the seizures of rats in CBZXD-Low group and CBZXD-High group were significantly reduced,and the damage to hippocampal neurons was alleviated,RORγt,IL-17A mRNA levels in spleen tissue were decreased,Foxp3,CTLA-4 and GITR mRNA levels were increased,the propor-tion of Th17 cells and Th17/Treg were decreased,the proportion of Treg cell was increased,intestinal flora α/β diversity were in-creased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were decreased,the proportion of beneficial bacte-ria,such as Bacteroides,Lactobacillus,Prevotella,and Akkermansia were increased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levels in hippocampal tissue were decreased(P<0.05);and it showed a dose-dependent effect of Chaibei Zhixian Decoction(P<0.05).Conclusion:Chaibei Zhixian Decoction can restore intestinal flora and Th17/Treg cell immune balance in epileptic rats,it possibly plays a role by regulating mTOR/HIF-1α pathway.

Objective To detect the expression of lactate dehydrogenase A(LDHA),FoxP3,and CD4 in hepatocellular carcinoma(HCC)and to analyze the effect of bufalin on LDHA as well as on the differentiation and function of regulatory T(Treg)cells in the tumor microenvironment of HCC mice.Methods Immunohistochemical staining was performed to detect the difference in the expression of LDHA,FoxP3(Treg cell marker),and CD4 between the tumor and adjacent tissues from 91 HCC patients.Hepl-6 subcutaneous tumor-bearing mouse model was established.The mice were randomly divided into control,bufalin+pyruvic acid,oxalate,and bufalin groups(n=5 each group).The mass and volume of subcutaneous tumors were compared.Immunohistochemical staining was performed to detect the expression of LDHA,FoxP3,and CD4 in tumors of mice in each group.ELISA was used to detect the levels of transforming growth factor β(TGF-β)and interleukin-10(IL-10)in the serum of mice in each group to evaluate the immune function of Treg cells.Results Compared with adjacent tissues,HCC tissues exhibited significantly higher LDHA and FoxP3 expression and lower CD4 expression(P＜0.001).The results of the correlation analysis showed that LDHA was positively correlated with FoxP3 expression and negatively correlated with CD4 expression(P＜0.05).The experimental results of Hep1-6 subcutaneous tumor-bearing mice showed that the tumor volume was significantly smaller in the bufalin and oxalate groups than in the control group and was significantly larger in the bufalin+pyruvic acid group than in the bufalin group(P＜0.001).The results of immunohistochemical staining showed that the expression levels of LDHA and the percentage of FoxP3+cells in the bufalin and oxalate groups were significantly lower than those in the control group,and the percentage of CD4+cells in the bufalin group was significantly higher than that in the control group(P＜0.001).The expres-sion level of LDHA and the percentage of FoxP3+cells in the bufalin+pyruvic acid group were significantly higher than those in the bufalin group,while the percentage of CD4+cells was significantly lower than that in the bufalin group(P＜0.001).The results of ELISA showed that the levels of TGF-β and IL-10 in the bufalin and oxalate groups were significantly lower than those in the control group(P＜0.001).Conclusion Bufalin inhibits cell growth,downregulates LDHA expression,and suppresses the differentiation and proliferation of Treg cells in HCC mice.

Objective:To investigate expressions of LDHA,CD56 and NKG2D in liver cancer tissues,and to further explore effects and correlations of Bufalin(BF)on LDHA and NK cells in hepatocellular carcinoma.Methods:Immunohistochemistry(IHC)was used to detect differential expressions of LDHA and NK cell specific molecular markers CD56 and NKG2D in 91 postoperative paraffin pathological specimens of hepatocellular carcinoma,as well as correlation between these markers in cancer and adjacent tissues.A Hepa1-6 subcutaneous tumor bearing hepatocellular carcinoma model was constructed by C57BL/6 mice and randomly divide into Control(Ctrl)group,BF+Pyruvic acid(BF+PA)group,Oxamate(OX)group,BF group,with 5 mice per group.Growth rate and volume of subcutaneous tumors were observed;IHC was used to detect expressions of LDHA and mouse NK cell specific molecular markers NK1.1 and NKG2D in tumors of each group of mice;ELISA was used to detect concentrations of perforin,TNF-α and IFN-γ in serum of mice in each group.Results:LDHA was highly expressed in human liver cancer tissues(P<0.001),while CD56 and NKG2D were highly expressed in adjacent tissues(P<0.001);LDHA expression was negatively correlated with CD56(r=-0.529 8,P<0.000 1)and NKG2D(r=-0.320 1,P<0.001);CD56 expression was positively correlated with NKG2D(r=0.612 2,P<0.000 1).Subcutaneous tumor experiment of mouse hepatocellular carcinoma showed that compared with Ctrl group and BF+PA group,subcutaneous tumor growth rate of OX group and BF group slowed down and showed a trend of tumor reduction;IHC detection showed that compared with Control group,LDHA expression in BF group was significantly downregulated(P<0.01),NK1.1 and NKG2D expressions were significantly increased(P<0.000 1);ELISA showed that compared with Ctrl group,LDHA in serum of BF group increased perforin,TNF-α,IFN-γ expressions(P<0.000 1),and enhanced NK cell killing ability.Conclusion:BF can inhibit LDHA expression in hepatocellular carcinoma cells and increase infiltration rate of NK cells in hepatocellular carcinoma tissues,which may enhance killing ability of NK cells in tumor microenvironment and inhibit growth of hepatocellular carcinoma.

Objective To explore the epidemic characteristics of common pneumonia pathogens in preschool children in Xining area and analyze the influencing factors of progression to severe pneumonia. Methods A total of 522 preschool children with pneumonia who were treated in our hospital from May 2021 to March 2024 were retrospectively selected as the research subjects. Sputum samples from children were taken to identify the pathogens and analyze their pathogenic epidemic characteristics.According to the diagnostic criteria in the 2019 version of ^“Standards for the Diagnosis and Treatment of Community-Acquired Pneumonia in Children^”, determine whether it is severe pneumonia, and collect the clinical data of the children.Logistic regression was used to analyze the influencing factors of the progression of common pneumonia to severe pneumonia. Results Among the 522 children with pneumonia, 522 cases were infected with pathogens, of which 447 cases were single infection (85.63%), 75 cases were mixed infection (14.36%). A total of 597 pathogens were detected, including 257 viruses (43.05%), 240 bacteria (40.20%), 68 mycoplasma pneumoniae (11.39%) and 32 chlamydia pneumoniae (5.36) . The detection rates of Streptococcus pneumoniae (149, 24.96%) and respiratory syncytial virus (118, 19.77%) were higher. Logistic regression results showed that length of hospital stay (OR=2.235, 95% CI: 1.552-3.439), ICU admission (OR=2.426, 95% CI: 1.769-3.881), intestinal microbiota disorder (OR=1.626, 95% CI: 1.335-2.842), multi-drug resistance (OR=2.086, 95%CI 1.417-2.905), mixed infection (OR=3.134, 95% CI : 2.217-8.857), nutritional risk (OR=2.783, 95% CI: 2.038-4.764), CRP (OR=2.589, 95% CI: 1.805-4.117), PCT (OR=1.486, 95%CI: 1.077-1.649), and white blood cells (OR=1.329, 95% CI: 1.021-1.536) were all associated with the risk of severe pneumonia (P<0.05). Conclusion The main pathogens of pneumonia in preschool children in Xining are Streptococcus pneumoniae and respiratory syncytial virus. Paying attention to the treatment of children with intestinal disorders, multiple infections, and malnutrition is of great significance to improve the progression of pneumonia.

Objective:To investigate expressions of LDHA,CD56 and NKG2D in liver cancer tissues,and to further explore effects and correlations of Bufalin(BF)on LDHA and NK cells in hepatocellular carcinoma.Methods:Immunohistochemistry(IHC)was used to detect differential expressions of LDHA and NK cell specific molecular markers CD56 and NKG2D in 91 postoperative paraffin pathological specimens of hepatocellular carcinoma,as well as correlation between these markers in cancer and adjacent tissues.A Hepa1-6 subcutaneous tumor bearing hepatocellular carcinoma model was constructed by C57BL/6 mice and randomly divide into Control(Ctrl)group,BF+Pyruvic acid(BF+PA)group,Oxamate(OX)group,BF group,with 5 mice per group.Growth rate and volume of subcutaneous tumors were observed;IHC was used to detect expressions of LDHA and mouse NK cell specific molecular markers NK1.1 and NKG2D in tumors of each group of mice;ELISA was used to detect concentrations of perforin,TNF-α and IFN-γ in serum of mice in each group.Results:LDHA was highly expressed in human liver cancer tissues(P<0.001),while CD56 and NKG2D were highly expressed in adjacent tissues(P<0.001);LDHA expression was negatively correlated with CD56(r=-0.529 8,P<0.000 1)and NKG2D(r=-0.320 1,P<0.001);CD56 expression was positively correlated with NKG2D(r=0.612 2,P<0.000 1).Subcutaneous tumor experiment of mouse hepatocellular carcinoma showed that compared with Ctrl group and BF+PA group,subcutaneous tumor growth rate of OX group and BF group slowed down and showed a trend of tumor reduction;IHC detection showed that compared with Control group,LDHA expression in BF group was significantly downregulated(P<0.01),NK1.1 and NKG2D expressions were significantly increased(P<0.000 1);ELISA showed that compared with Ctrl group,LDHA in serum of BF group increased perforin,TNF-α,IFN-γ expressions(P<0.000 1),and enhanced NK cell killing ability.Conclusion:BF can inhibit LDHA expression in hepatocellular carcinoma cells and increase infiltration rate of NK cells in hepatocellular carcinoma tissues,which may enhance killing ability of NK cells in tumor microenvironment and inhibit growth of hepatocellular carcinoma.

Objective To detect the expression of lactate dehydrogenase A(LDHA),FoxP3,and CD4 in hepatocellular carcinoma(HCC)and to analyze the effect of bufalin on LDHA as well as on the differentiation and function of regulatory T(Treg)cells in the tumor microenvironment of HCC mice.Methods Immunohistochemical staining was performed to detect the difference in the expression of LDHA,FoxP3(Treg cell marker),and CD4 between the tumor and adjacent tissues from 91 HCC patients.Hepl-6 subcutaneous tumor-bearing mouse model was established.The mice were randomly divided into control,bufalin+pyruvic acid,oxalate,and bufalin groups(n=5 each group).The mass and volume of subcutaneous tumors were compared.Immunohistochemical staining was performed to detect the expression of LDHA,FoxP3,and CD4 in tumors of mice in each group.ELISA was used to detect the levels of transforming growth factor β(TGF-β)and interleukin-10(IL-10)in the serum of mice in each group to evaluate the immune function of Treg cells.Results Compared with adjacent tissues,HCC tissues exhibited significantly higher LDHA and FoxP3 expression and lower CD4 expression(P＜0.001).The results of the correlation analysis showed that LDHA was positively correlated with FoxP3 expression and negatively correlated with CD4 expression(P＜0.05).The experimental results of Hep1-6 subcutaneous tumor-bearing mice showed that the tumor volume was significantly smaller in the bufalin and oxalate groups than in the control group and was significantly larger in the bufalin+pyruvic acid group than in the bufalin group(P＜0.001).The results of immunohistochemical staining showed that the expression levels of LDHA and the percentage of FoxP3+cells in the bufalin and oxalate groups were significantly lower than those in the control group,and the percentage of CD4+cells in the bufalin group was significantly higher than that in the control group(P＜0.001).The expres-sion level of LDHA and the percentage of FoxP3+cells in the bufalin+pyruvic acid group were significantly higher than those in the bufalin group,while the percentage of CD4+cells was significantly lower than that in the bufalin group(P＜0.001).The results of ELISA showed that the levels of TGF-β and IL-10 in the bufalin and oxalate groups were significantly lower than those in the control group(P＜0.001).Conclusion Bufalin inhibits cell growth,downregulates LDHA expression,and suppresses the differentiation and proliferation of Treg cells in HCC mice.

Objective:To explore the effect of Chaibei Zhixian Decoction on intestinal flora and T helper 17/regulatory T cell(Th17/Treg)immune balance in epileptic rats by regulating mammalian target of rapamycin/hypoxia-inducible factor-1α(mTOR/HIF-1α)pathway.Methods:Sixty rats were randomly divided into Sham group,Model group,low-dose Chaibei Zhixian Decoction group(CBZXD-Low)and high-dose Chaibei Zhixian Decoction group(CBZXD-High),with 15 rats in each group.Lateral ventricular injec-tion 1 μl kainic acid(1.5 μg/μl)was used to prepare epileptic rat model,Sham group was injected with an equal amount of physiological saline.CBZXD-Low group and CBZXD-High group were given Chaibei Zhixian Decoction by gavage daily(8.48 g/kg,16.96 g/kg),while Sham group and model group were given an equal amount of physiological saline by gavage.After continuous gavage for 4 weeks,HE staining and Nissl staining were used to detect pathological damage in hippocampal tissue;RT-qPCR was used to detect Th17/Treg related marker mRNA levels in spleen tissue and mTOR,HIF-1α mRNA levels in hippocampus tissue;flow cytometry was used to de-tect Th17/Treg cell ratio in spleen tissue;16S rRNA sequencing of feces in colon segment was performed;Western blot was used to de-tect mTOR and HIF-1α protein levels in hippocampal tissue.Results:Compared with Sham group,Model group rats suffered from spontaneous epilepsy,the arrangement of hippocampal neurons was disordered and severely lacking,intracellular Nissl bodies was de-creased,RORγt,IL-17A mRNA levels in spleen tissue were increased,Foxp3,CTLA-4 and GITR mRNA levels were decreased,the proportion of Th17 cells and Th17/Treg were increased,the proportion of Treg cell was decreased,intestinal flora α/β diversity were decreased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were increased,the proportion of beneficial bacte-ria such as Bacteroides,Lactobacillus,Prevotella and Akkermansia were decreased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levelsin hippocampal tissue were increased(P<0.05);compared with Model group,the seizures of rats in CBZXD-Low group and CBZXD-High group were significantly reduced,and the damage to hippocampal neurons was alleviated,RORγt,IL-17A mRNA levels in spleen tissue were decreased,Foxp3,CTLA-4 and GITR mRNA levels were increased,the propor-tion of Th17 cells and Th17/Treg were decreased,the proportion of Treg cell was increased,intestinal flora α/β diversity were in-creased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were decreased,the proportion of beneficial bacte-ria,such as Bacteroides,Lactobacillus,Prevotella,and Akkermansia were increased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levels in hippocampal tissue were decreased(P<0.05);and it showed a dose-dependent effect of Chaibei Zhixian Decoction(P<0.05).Conclusion:Chaibei Zhixian Decoction can restore intestinal flora and Th17/Treg cell immune balance in epileptic rats,it possibly plays a role by regulating mTOR/HIF-1α pathway.

Objective To investigate the methylation levels of the HIF-3α gene,as well as the expression of HIF-3α mRNA and DDIT4 mRNA in peripheral blood samples from pregnant women with gestational diabetes mellitus(GDM),and their association with pregnancy outcomes,aiming to provide insights for pregnancy monitoring and clinical diagnosis and treatment of GDM patients,thereby mitigating the risk of adverse pregnancy outcomes.Methods From March 2023 to March 2024,a total of 80 pregnant women with GDM were randomly selected at the Affiliated Hospital of Xuzhou Medical University.They were divided into two groups based on glycemic control:GDM1(n=40)representing good glycemic control and GDM2(n=40)representing poor glycemic control.Addi-tionally,a control group consisting of 40 pregnant women without any pregnancy complications or comorbidities was included.Methylation-specific PCR was employed to determine the rate of HIF-3α methylation,while quantita-tive real-time PCR was used to assess the expressions of HIF-3α mRNA and DDIT4 mRNA.Adverse pregnancy outcomes were recorded across all three groups,and correlations between HIF-3α mRNA,DDIT4 mRNA,and various adverse pregnancy outcomes were analyzed using logistic regression analysis.Results Prior to pregnancy,GDM1 and GDM2 groups exhibited higher levels of FPG,HbA1c,TC,TG,LDL-C,FINS and HOMA-IR(P＜0.05),as well as lower levels of HOMA-β compared to the control group(P＜0.05).The incidence of adverse pregnancy outcomes was also higher in both GDM groups than in the control group;this difference was significant for GDM2(P＜0.05)but not for GDM1(P＞0.05).Furthermore,methylation rates of the HIF-3α gene were higher in both GDM groups compared to controls while expression levels of HIF-3α and DDIT4 mRNA were lower;these differences were significant for the GDM2 group versus controls(P＜0.05)but not for the GDM1 group ver-sus controls(P＞0.05).Finally,all adverse pregnancy outcomes among women with gestational diabetes mellitus showed a negative correlation with expression levels of both HIF-3α and DDIT4 mRNA(r＜0,P＜0.05);moreover,these two mRNAs served as protective factors against such outcomes occurring(OR＜1,P＜0.05).Conclusion The higher methylation rate of HIF-3α and the decreased expression of HIF-3α and DDIT4 genes are associated with the development of GDM and may serve as significant factors contributing to adverse pregnancy outcomes.