Objective To investigate the effect of Hedyotis diffusa extract(HDE)on the proliferation of liver cancer cells and its relationship with sugar metabolism reprogramming and oxidative phosphorylation and analyze its possible mechanisms.Methods CCK-8 and EDU experiments were used to determine the effect of different concentrations(20,40,80 mg/mL)of HDE on the growth of liver cancer cell line SNU-368.Lactate dehydrogenase activity,glucose uptake,lactate production,extracellular pH,mitochondrial respiratory chain complex activity,and cellular oxygen consumption were measured to analyze the effect of HDE on aerobic glycolysis and oxidative phosphorylation in liver cancer cells.qRT-PCR experiments were used to detect the mRNA expressions of GLUT1,GLUT4,HK2,GPI,PFKL,ALDOA and HIF-1α in SNU-368 cells of different groups.Western blotting experiments were used to detect the protein expression of HIF-1α.A stable cell line overexpressing HIF-1αwas constructed by lentivirus transfection of liver cancer cells SNU-368 and then intervened with HDE;the expression of HIF-1α mRNA and protein was detected with qRT-PCR and Western blotting.Results CCK-8 results showed that the HDE exhibited a concentration-dependent inhibitory effect on the proliferation of liver cancer cells(all P＜0.05).Results from glucose metabolism-related tests indicated that the HDE could inhibit glucose uptake and lactate production,decrease lactate dehydrogenase activity,increase extracellular pH value,enhance cellular oxygen consumption,and elevate activities of mitochondrial respiratory chain complexes Ⅰ,Ⅱ,Ⅲ and Ⅳ(all P＜0.05).qRT-PCR results revealed that the HDE suppressed the mRNA expressions of GLUT1,HK2,GPI,and ALDOA(all P＜0.05).qRT-PCR and Western blotting experiments showed that compared to the control group,the expression of HIF-1α mRNA and protein in the HDE group was significantly reduced.However,when HIF-1α was overexpressed and HDE was added in the HIF-1α-LV group,the expression of HIF-1α mRNA and protein increased again compared to the HDE group.Conclusion HDE inhibits glycolysis and promotes oxidative phosphorylation to inhibit the proliferation of liver cancer cells,and its mechanism of action may be related to the inhibition of HIF-1α expression.

Objective To investigate the effect and mechanism of Twist-related protein 1(TWIST1)on pulmonary vascular remodeling induced by monocrotaline(MCT)in pulmonary arterial hypertension(PAH)rats.Methods A total of 50 healthy male Sprague Dawley(SD)rats were randomly divided into five groups including control group,MCT-treated group,MCT and dimethyl sulfoxide(DMSO)-treated group,MCT and harmine-treated group MCT and hydroxychloroquine(HCQ)-treated group.The right ventricle systolic pressure(RVSP)was measured,right ventricular hypertrophy index(RVHI)and percentage of medial wall thickness(MT％)to assess the development of PAH.The protein levels of TW1ST1,autophagy markers LC3B and RND3 were determined using western blot.Results Compared with control group,expressions of TWIST1 and LC3B were increased by 2.32±0.22 folds and 0.87±0.19 folds in MCT-induced PAH group,with significant differences(t=15.812,11.227,all P＜0.00 1),while the protein level of RND3 in MCT-induced PAH rats was decreased by 0.32±0.07 folds compared with control group,with significant difference(t=-13.003,P＜0.001).Administration of TWIST1 inhibitor Harmine or autophagy inhibitor hydroxychloroquine significantly suppressed MCT-induced increase in LC3B and down-regulation of RND3 expression,and reduced RVSP,RVHI and MT％expressions in MCT-induced PAH rats,with significant differences(t=-24.277～16.636,all P＜0.001).Conclusion TWIST1 promotes pulmonary vascular remodeling by inducing autophagy activation,thus promoting the occurrence and development of PAH.

COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.

Objective To evaluate the clinical efficacy of single-session plasmapheresis therapy alone for the treatment of toxic epidermal necrolysis (TEN),and to investigate its adverse reactions.Methods Patients with TEN receiving single-session plasmapheresis therapy alone were collected from the Second Affiliated Hospital of Xi'an Jiaotong University between September 2010 and December 2017.Clinical data on the disease severity,clinical efficacy,hospitalization duration and adverse reactions were analyzed.Results A total of 17 patients with TEN were enrolled into this study,including 9 males and 8 female,with an average age of 36.1 ± 25.4 years.Their initial SCORTEN and STENS scores were 2.1 ± 1.24 and 29.9 ± 6.6 respectively.After treatment,the STENS score decreased to 3.5 ± 1.8.Of the 17 patients,15 were cured after single-session plasmapheresis therapy,1 showed response to the treatment,and 1 died.The duration of intensive care unit stay was 6.4 ± 1.8 days,and the total hospitalization duration was 12.1 ± 5.7 days.There was no significant difference in the STENS score among the day 1,4,7,10 and 20 after hospital admission (F =18.569,P ＜ 0.05).No severe adverse reactions were observed,except 2 cases of plasma allergy.Conclusion Single-session plasmapheresis therapy alone is effective for the treatment of TEN without obvious adverse reactions.

Objective To explore the diagnostic and score value of ultrasound on hemophiliac arthropathy referring to MRI on the diagnosis and score of hemophiliac arthropathy Methods The ultrasound and MRI examinations were performed on 42 joints of 42 hemophilia patients 14 knees 14 ankles and 14 elbows The consistency of ultrasound and magnetic resonance imaging in the detection and score of joint diseases was compared Finally inter-and intra-observer agreement of ultrasound scoring system were tested Results The consistency of ultrasound and magnetic resonance imaging was excellent κ=0 763-0 896 P < 0 001 in the detection of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy hemosiderin excellent κ=0 793 P <0 001 in the detection of cartilage loss poor κ=0 133 P = 0 132 in the detection of erosions and poor κ= 0 100 P = 0 137 in the detection of subchondral cysts The consistency of ultrasound and magnetic resonance imaging was good to excellentκ=0 684-0 833 P < 0 001 in the score of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy and hemosiderin and poor to good κ=0 145 -0 635 P <0 001 in the score of advanced osteochondral lesions cartilage loss and bone erosions The inter-observer agreement was good to excellent κ=0 676-0 870 P <0 001 for early soft tissue lesions and moderate to excellent κ=0 421- 0 75 1 P < 0 001 for advanced osteochondral lesions The intra-observer agreement was good to excellent κ=0 705-0 885 P <0 001 for early soft tissue lesions and moderate to good κ=0 532 -0 732 P <0 001 for advanced osteochondral lesions Conclusions Ultrasound plays an important role in detecting early soft tissue changes effusion or hemarthrosis synovial hypertrophy hemosiderin and cartilage loss which helps follow-up and guide clinical treatment.

[ ABSTRACT ] AIM: To investigate the effect of catalpol on the activity of osteoblasts ( OB ) and osteoclasts ( OC) , and OB estrogen receptor ( ER) α/βmRNA expression in the OB-OC co-culture system.METHODS: OB and OC were isolated from the SD rats of 1 and 5 days old.In the OB-OC co-culture system, different concentrations of catalpol including low dosage (0.05 , 0.1, 0.5 and 1 mg/L), middle dosage (2, 5 and 10 mg/L), and high dosage (20, 50 and 100 mg/L) were added into the culture medium to detect the changes of OB proliferation by MTT assay.The catalpol at maximal dosage was added to OB section to detect the alkaline phosphatase ( ALP) activity of OB by pNPP method.The mRNA expression of ERα/βin the OB treated with catalpol in the co-culture system was detected by RT-PCR.The catalpol at maximal dosage was added to OC group to detect the activity of OC by microscopy and tartrate-resistantacid phosphatase ( TRAP) activity detection.RESULTS:In 0.05~2 mg/L catalpol groups, the proliferation of OB was significantly in-creased as compared with control group in the co-culture system, and it reached the maximum value when catalpol was at 0.05 mg/L, while in 5~100 mg/L catalpol groups, the proliferation of OB was not increased.The ALP activity of OB in 0.05 mg/L catalpol group was higher than that in control group.The catalpal at 0.05 mg/L promoted the mRNA expression of ERβin OB in the co-culture system, but did not increase the mRNA expression of ERαas compared with control group. Catalpol at 0.05 mg/L obviously inhibited the bone resorption and the TRAP activity in OC.CONCLUSION: Catalpol stimulates the proliferation and activity of OB, inhibits the bone resorption and activity of OC, and increases the mRNA ex-pression of ERβin OB in the OB-OC co-culture system, suggesting that high mRNA expression of ERβmay be the regula-tory pathway of catalpol in response to bone metabolism.

OBJECTIVETo systematically review whether statins can reduce the risk of infection and infection-related mortality.

METHODSWe searched the Cochrane Library, MEDLINE, EMBASE, PubMed, Elsevier and CBM databases for randomized placebo-controlled trials of statins published by September 2013, and each trial enrolled at least 100 participants with follow-up for at least 4 weeks. Two reviewers independently assessed the quality of the included studies and extracted the relevant data for analysis using Stata 12.0 software.

RESULTSSixteen trails involving a total of 48973 patients were included in our meta-analysis. The results showed that statins significantly reduced the risk of infection (OR=0.93, 95% CI 0.89 to 0.98, P=0.004) compared to placebo but did not significantly lower infection-related mortality (OR=0.96, 95% CI 0.82 to 1.12, P=0.592).

CONCLUSIONStatins can significantly reduce the risk of infection but does not lower infection-related mortality.

Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; therapeutic use ; Infection ; epidemiology ; mortality ; Randomized Controlled Trials as Topic ; Risk Factors

OBJECTIVETo construct a humulus pollen allergy DNA vaccine pcDNA3.1-Hum and investigate its effect for immune protection mediated by Foxp3(+)Treg cells in asthmatic mice.

METHODSThe target humulus gene obtained from pTripIEx2-Hum plasmid by double enzyme digestion was inserted sequentially into pcDNA3.1(-) vector to generate the recombinant plasmid pcDNA3.1-Hum, which was validated by sequencing. The pcDNA3.1-Hum plasmid was transfected into COS-7 cells and the expression of the ectopic protein was analyzed using Western blotting. Co-cultured dendritic cells and CD4(+)CD25(-) T cells were stimulated with the expressed protein to test its efficacy in inducing Foxp3(+)Treg cells. The levels of humulus-specific IgE and IgG2a were assayed to evaluate the allergenicity and immunogenicity of pcDNA3.1-Hum in mice. The immunoprotective effect of pcDNA3.1-Hum was assessed in a mouse model of humulus-specific asthma.

RESULTSThe constructed pcDNA3.1-Hum plasmid was validated by sequencing and Western blotting, and the expressed protein was shown to induce Foxp3(+)Treg cells in the co-culture. In normal mice, pcDNA3.1-Hum induced a significant increase of humulus-specific IgG2a but had no effect on IgE. In the asthmatic mice, pcDNA3.1-Hum significantly decreased inflammatory cell counts and eosinophil percentages in the BALF, ameliorated lung inflammation, and lowered AHR and IL-4 levels; immunization of the mice with pcDNA3.1-Hum reversed humulus-induced reduction of serum IFN-γ and prevented the humulus-triggered reduction of Foxp3(+)Treg cell percentage in the spleen.

CONCLUSIONWe have successfully constructed a highly immunogenic pcDNA3.1-Hum DNA vaccine that can mediate immune protection by inducing Foxp3(+)Treg cells.

Allergens ; immunology ; Animals ; Asthma ; immunology ; Cell Differentiation ; Disease Models, Animal ; Female ; Humulus ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmids ; T-Lymphocytes, Regulatory ; cytology ; immunology ; Vaccines, DNA ; immunology

Objective To systematically review whether statins can reduce the risk of infection and infection-related mortality. Methods We searched the Cochrane Library, MEDLINE, EMBASE, PubMed, Elsevier and CBM databases for randomized placebo-controlled trials of statins published by September 2013, and each trial enrolled at least 100 participants with follow-up for at least 4 weeks. Two reviewers independently assessed the quality of the included studies and extracted the relevant data for analysis using Stata 12.0 software. Results Sixteen trails involving a total of 48973 patients were included in our meta-analysis. The results showed that statins significantly reduced the risk of infection (OR=0.93, 95% CI 0.89 to 0.98, P=0.004) compared to placebo but did not significantly lower infection-related mortality (OR=0.96, 95% CI 0.82 to 1.12, P=0.592). Conclusion Statins can significantly reduce the risk of infection but does not lower infection-related mortality.

Objective To construct a humulus pollen allergy DNA vaccine pcDNA3.1-Hum and investigate its effect for immune protection mediated by Foxp3+Treg cells in asthmatic mice. Methods The target humulus gene obtained from pTripIEx2-Hum plasmid by double enzyme digestion was inserted sequentially into pcDNA3.1(-) vector to generate the recombinant plasmid pcDNA3.1-Hum, which was validated by sequencing. The pcDNA3.1-Hum plasmid was transfected into COS-7 cells and the expression of the ectopic protein was analyzed using Western blotting. Co-cultured dendritic cells and CD4+CD25-T cells were stimulated with the expressed protein to test its efficacy in inducing Foxp3+Treg cells. The levels of humulus-specific IgE and IgG2a were assayed to evaluate the allergenicity and immunogenicity of pcDNA3.1-Hum in mice. The immunoprotective effect of pcDNA3.1-Hum was assessed in a mouse model of humulus-specific asthma. Results The constructed pcDNA3.1-Hum plasmid was validated by sequencing and Western blotting, and the expressed protein was shown to induce Foxp3+Treg cells in the co-culture. In normal mice, pcDNA3.1-Hum induced a significant increase of humulus-specific IgG2a but had no effect on IgE. In the asthmatic mice, pcDNA3.1-Hum significantly decreased inflammatory cell counts and eosinophil percentages in the BALF, ameliorated lung inflammation, and lowered AHR and IL-4 levels; immunization of the mice with pcDNA3.1-Hum reversed humulus-induced reduction of serum IFN-γ and prevented the humulus-triggered reduction of Foxp3+Treg cell percentage in the spleen. Conclusion We have successfully constructed a highly immunogenic pcDNA3.1-Hum DNA vaccine that can mediate immune protection by inducing Foxp3+Treg cells.