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 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.

The outbreak of COVID-19 has spread quickly across 114 countries/territories/areas in six continents worldwide and has been announced as a pandemic by WHO. This study analyzed global COVID-19 epidemiological trends, examined impact of the pandemic on global health security, diplomacy, and social environment in China, and provided short- and long-term strategic policy recommendations for China’s subsequent preparedness and responses.

Objective To investigate the effect of DPP-4 inhibitor sitagliptin on atherosclerosis and its mechanism.Methods Thirty male ApoE-/-mice were randomly divided into two groups:experimental group(n=15) and control group(n=15).The mice in the experimental group were fed with high-fat mixture of sitagliptin and the control group was fed with high fat.Collected blood in the eyeballs in order to analyze serum levels of blood lipids and blood glucose after 16 weeks of feeding,and detected serum nitric oxide synthase(eNOS),vascular adhesion molecule-1(VCAM-1) with ELISA method.Collected aortic tissue in order to analyze atherosclerotic plaque.Results There was no significant difference in blood glucose,triglyceride and total cholesterol level between the two groups(P>0.05).The serum high density lipoprotein in the experimental group was significantly higher than that in the control group(P<0.05).The atherosclerotic plaque in the experimental group(7.55±1.87)%, which was significantly smaller than that in the control group(11.67±1.32)%.The serum VCAM-1 in the experimental group was lower and the eNOS was higher than that in the control group(P<0.05).Conclusion DPP-4 sitagliptin can increase the expression of HDL and eNOS and inhibit the expression of VCAM-1,thereby inhibiting the progression of atherosclerosis in ApoE-/-mice.