Objective To compare ablation ranges of single and double needle microwave ablation(MWA)of pig lung in vivo.Methods Five healthy Bama miniature pigs were enrolled.Single needle(single needle group)and parallel double needle MWA(double needle group)were performed successively on bilateral lungs,respectively.Adverse event during MWA was evaluated according to common terminology criteria for adverse events(CTCAE).The long diameter(D1CT),short diameter(D2CT),longitudinal diameter(D3CT)and volume(V)of ablation foci were measured and calculated based on CT images immediately after MWA,whereas D1,s D2s and sphericity of ablation foci were also obtained based on specimen and compared between groups.Results D1CT,D2CT,D3CT and V,as well as D1S,D2S and sphericity of ablation foci in single needle group were all significantly smaller than those in double needle group(all P＜0.01).Mild pneumothorax(CTCAE grade 1)was found in 1 pig(1/5,20.00%)in single needle group,while mild pneumothorax and pulmonary hemorrhage(both CTCAE grade 1)occurred in 1 pig(1/5,20.00%)in double needle group.No other adverse event was observed.Conclusion Compared with single needle MWA,double needle MWA of pig lung in vivo resulted larger ablation range and more spherically shaped ablation foci.

OBJECTIVE: Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro. METHODS: The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining. RESULTS: The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism. CONCLUSION Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

Objective To investigate the level of coagulation factor Ⅺ(FⅪ)in patients with venous thrombosis of lower limbs and its correlation with recurrence risk.Methods A total of 220 pa-tients with deep vein thrombosis(DVT)admitted in our hospital from February 2018 to February 2019 were enrolled as the study group,and another 50 healthy individuals taking physical exami-nation during same period served as the control group.After a 3 years followed,the study group ultimately included 197 cases,according to the results of restricted cubic spline(RCS),the study group was divided into low(FⅪ＜10.3 U/L,94 cases),medium-(10.3-12.1 U/L,52 cases)and high-level groups(＞12.1 U/L,51 cases).The plasma level of FⅪ was detected in the study group 1 month after the end of anticoagulant therapy,and the results were compared with those of the control group during physical examination.Cox model was used to analyze the influence of FⅪ on the recurrence of DVT,and RCS was employed to analyze the relationship between DVT recur-rence and FⅪ level.Kaplan-Meier curve was plotted to analyze the recurrence risk of DVT with different FⅪ levels.The patients from the study group were followed up for 3 years.Results The FⅪ level was significantly higher in the study group than the control group(P＜0.05).During fol-low-up period,33 patients(16.75％)had DVT recurrence.The Cox model analysis after adjust-ment of sex and age showed that FⅪ level was a risk factor for DVT recurrence(P＜0.05).When the FⅪ level was set into tertile and the risk ratio was calculated after adjustment,FⅪ＜10.3 U/L,and the average FⅪ level at this stage was 9.2 U/L,the risk ratio was 0.82(95％CI:0.673-0.984);Patients with FⅪ between 10.3 and 12.1 U/L,and the average FⅪ at this stage was 11.4 U/L,the risk ratio of 1.04(95％CI:0.813-1.432).The those with FⅪ＞12.1 U/L,and the average FⅪ at this stage was 13.8 U/L,hazard ratio of 1.38(95％CI:0.921-1.563).Kaplan-Meier curve analysis showed that the recurrence risk was 28.62％(95％CI:25.633-31.609),30.10％(95％CI:27.594-32.606)and 38.06％(95％CI:34.306-41.371),respectively for the low-,medium-,and high-level groups,with significant correlation among the three groups(x2=6.631,P=0.036).Conclusion Compared with healthy individuals,plasma FⅪ level is at a high level in the DVT patients.With the increment of FⅪ level,the risk of DVT recurrence increases.Two FⅪ levels,10.3 U/L and 12.1 U/L,can be used as reference points for the obvious increase of DVT recur-rence rate.

ObjectiveTo observe the therapeutic effect of BD-77 by nebulized inhalation on animal models of various respiratory viral infections and investigate the mechanism of broad-spectrum antiviral action of BD-77 using proteomics. MethodThe influenza virus H1N1/FM1 experiment used ICR mice and divided them into a normal group， model group， Tamiflu group， and BD-77 groups of 75 and 37.5 g·L^-1 for inhalation of 20 min and 25 min. Human coronavirus 229E and OC43 experiment divided the BALB/c mice into a normal group， model group， chloroquine phosphate group， and BD-77 groups of 75， 37.5， 18.75， and 9.375 g·L^-1， with 10 mice in each group. Influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection-induced pneumonia models were used to detect mouse lung index， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the viral load in lung tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect related inflammatory factors in lung tissue， and proteomics analysis was performed on the lung tissue of OC43-infected mice. ResultCompared with that in the normal group， the lung index of mice in each infection group was significantly increased （P<0.01）， and viral nucleic acid could be detected in the lung tissue of mice infected with human coronaviruses 229E and OC43. The levels of interleukin-6 （IL-6）， IL-10， and tumor necrosis factor-α （TNF-α） in the lung tissue of mice infected with human coronavirus 229E were all significantly increased （P<0.01）. BD-77 could significantly reduce the lung index of mice infected with influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 （P<0.05， P<0.01）， cut down the viral load in the lungs of mice infected with human coronaviruses 229E and OC43 （P<0.01）， and lower the contents of IL-6， IL-10， and TNF-α in the lung tissue of mice infected with human coronavirus 229E （P<0.01）. Proteomics analysis of the lung tissue of OC43-infected mice showed that BD-77 regulated the AMPK signaling pathway， TNF signaling pathway， NOD-like signaling pathway， IL-17 signaling pathway， Forkhead box protein O （FoxO） signaling pathway， transforming growth factor-β （TGF-β） signaling pathway， and other signaling pathways. ConclusionNebulized inhalation of BD-77 is effective in treating pneumonia caused by influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection in mice and may exert its antiviral effects by regulating the balance of cellular metabolism， enhancing the immune function of the host， and attenuating inflammatory responses.

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.

OBJECTIVES: Long-term hepatitis B virus (HBV) infection can cause recurrent inflammation in the liver, and then develop into liver fibrosis, cirrhosis, and liver cancer. The hepatic pathological change is one of the important criteria for guiding antiviral therapy in patients with chronic hepatitis B (CHB). Due to the limitations of liver biopsy, it is necessary to find valuable non-invasive indicators to evaluate the hepatic pathological changes in CHB patients and guide the antiviral therapy. This study aims to analyze the clinical characteristics of different pathological changes in CHB patients, and to explore the factors influnencing the degree of liver inflammation and fibrosis in CHB patients with normal alanine aminotransferase (ALT). METHODS: This retrospective study was conducted on 310 CHB patients. Liver biopsy was performed in all these patients. The clinical data of the patients were collected. The liver biopsy pathological results were used as the gold standard to analyze the relationship between clinical indicators and liver pathological changes. Then CHB patients with normal ALT were screened, and the independent factors influencing the degree of liver inflammation and fibrosis were explored. RESULTS: Among the 310 patients with CHB, there were 249 (80.3%) patients with significant liver inflammation [liver inflammation grade (G) ≥2] and 119 (38.4%) patients with significant liver fibrosis [liver fibrosis stage (S) ≥2]. The results of univariate analysis of total samples showed that the ALT, γ-glutamyl transferase, alkaline phosphatase, and HBV DNA were related to the significant liver pathological changes. Among the 132 CHB patients with normal ALT, the patients with liver pathology G/S≥2, G≥2, and S≥2 were 80.3% (106/132), 68.2% (90/132), and 43.2% (57/132), respectively. The results showed that the independent influencing factor of significant liver inflammation was HBV DNA>2 000 U/mL (OR=3.592, 95% CI 1.534 to 8.409), and the independent influencing factors of significant liver fibrosis were elevated alkaline phosphatase level (OR=1.022, 95% CI 1.002 to 1.043), decreased platelet count (OR=0.990, 95% CI 0.982 to 0.998), and positive in hepatitis B e antigen (HBeAg) (OR=14.845, 95% CI 4.898 to 44.995). According to the multivariate analysis, a diagnostic model for significant liver fibrosis in CHB patients with normal ALT was established, and the area under the receiver operating characteristic curve was 0.844 (95% CI 0.779 to 0.910). CONCLUSIONS The liver pathological changes should be evaluated in combination with different clinical indicators. A considerable number of CHB patients with normal ALT still have significant liver pathological changes, which need to be identified and treated with antiviral therapy in time. Among them, HBV DNA>2 000 U/mL suggests the significant liver inflammation, and the diagnostic model for significant liver fibrosis based on alkaline phosphatase, platelet count, and HBeAg can help to evaluate the degree of liver fibrosis.
Humans ; Hepatitis B, Chronic/complications* ; Hepatitis B e Antigens/therapeutic use* ; Alkaline Phosphatase ; DNA, Viral ; Retrospective Studies ; Fibrosis ; Hepatitis B virus/genetics* ; Liver Cirrhosis/etiology* ; Inflammation/drug therapy* ; Antiviral Agents/therapeutic use* ; Alanine Transaminase

Objective: To understand the epidemiological characteristics of foodborne disease outbreaks in schools in Zhejiang province, and to provide evidence for effective prevention and control of foodborne disease outbreaks in schools. Methods: A descriptive analysis was conducted on foodborne disease outbreaks in Zhejiang schools reported by the national foodborne disease outbreaks surveillance system from 2010 to 2019. Results: During the past 10 years, a total of 86 foodborne disease outbreaks in schools were reported, with 1 755 illnesses, 240 hospitalizations, and no deaths. Pathogenic bacteria and their toxins were the main causes of foodborne disease outbreaks in schools, accounting for 83.0%(44/53) of all identified causes. The top four types of pathogenic bacteria were Staphylococcus aureus, Salmonella, diarrheagenic Escherichia coli, and Bacillus cereus. Meat products and mixed foods were the main foods that caused the outbreaks, each accounting for 16.3%(14/86) of total incidents. High school cafeterias were places with the highest incidence, accounting for 38.4%(33/86) of the total. School concession stands caused the largest number of hospitalizations, accounting for 37.1%(89/240) of the total. The peak month of foodborne disease outbreaks in schools was September, followed by June, May, and October. Crosscontamination and improper storage were the main causes of foodborne disease outbreaks in schools. Conclusion Bacterial foodborne disease is a major food safety issue in schools in Zhejiang Province. In summer and fall, school cafeterias and food stores should take effective measures to prevent bacterial foodborne disease outbreaks caused by cross-contamination and improper storage of high-risk foods such as meat products and cold-processed bakeries.

Objective To evaluate the clinical efficacy of prophylactic cranial irradiation (PCI) in the treatment of surgically resected small cell lung cancer (SCLC).Methods Clinical data of SCLC patients undergoing radical resection surgery in Zhejiang Cancer Hospital from 2003 to 2015 were retrospectively analyzed.According to the treatment modality,all patients were allocated into the PCI and non-PCI groups.A total of 52 patients were finally included,including 19 patients in the PCI group (5 cases of stage Ⅰ,5 stage Ⅱ and 9 stage Ⅲ) and 33 in the non-PCI group (12 cases of stage Ⅰ,5 stage Ⅱ and 16 stage Ⅲ).Kaplan-Meier method was utilized for survival analysis.Cox proportional hazards model was adopted to analyze clinical prognosis.Results The median survival time was 32.9 months in the PCI group,and 20.4 months in the non-PCI group.The 2-year overall survival rate was 72％ in the PCI group,significantly higher than 38％ in the non-PCI group (P=0.023).The median brain metastasis-free survival (BMFS) was 32.5 months in the PCI group,and 17.1 months in the non-PCI group.In the PCI group,the 2-year BMFS rate was 89％,significantly better than 53％ in the non-PCI group (P=0.026).Subgroup analysis demonstrated that PCI could confer survival benefit to patients with p-stage Ⅲ (p=0.031) rather than p-stage Ⅰ (P=0.924) and Ⅱ (P=0.094) counterparts.Multivariate analysis revealed that PCI (HR=0.330,P=0.041) was an independent prognostic factor of the overall survival.Conclusions PCI can reduce thr risk of brain metastasis rate and improve the overall survival of patients with surgically resected SCLC.

This article was aimed to study the effect of polyphenols fromRubus suavissirnusS. Lee (RSLP) on spontaneous hypertensive rats (SHR) and to explore its mechanism of anti-hypertensive. The water extraction of RSLP was prepared. And the polyphenols was extracted with macroporous resin. The non-invasive blood pressure analysis system was used to detect the blood pressure. SHR model was selected to study the anti-hypertensive effect. The 16 normal Wistar rats were randomly divided into the control group and the normal RSLP high-dose group (RSLP-NH). The 40 SHR were randomly divided into the model group, Captopril group, RSLP-L group, RSLP-M group and RSLP-H group. SBP, DBP, HR, body weight and organ index were observed after the drug administration for 8 weeks and drug withdrawal for 2 weeks. The contents of SOD, MDA, GSH-Px, NO, NOS and ANP in serum were measured. The results showed that the blood pressure of SHR was significantly higher than that of the control group, which can be used for anti-hypertensive studies. Each RSLP group can obviously reduce the SBP and DBP of SHR (P < 0.05), but it had no effect on HR (P > 0.05). RSLP can elevate GSH-Px, SOD levels and reduce the activity of MDA (P < 0.05). RSLP can reduce NO, NOS and ANP contents in serum (P < 0.05). It was concluded that RSLP can significantly reduce the SBP and DBP of SHR, but it had no significant effect on HR. It can increase the activity of GSH-Px, SOD, NO, NOS levels, and reduce the contents of MDA, ANP in serum. It had certain inhibitory effect on the left ventricular hypertrophy.

Objective:To investigate the anti-proliferation effect of carvacrol on human gastric cancer cell BGC-823 and explore the molecular mechanism. Methods:The proliferation of BGC-823 cells was evaluated by MTT assay. Flow cytometry was used to ana-lyze the cell apoptosis after exposed to carvacrol. Transwell assay was used to analyze the effect of carvacrol on cell metastasis. Quanti-tative realtime-PCR was used to detect the expression of MMP-9 and TIMP-1. The expression of caspase-9 and PARP,and the activa-tion of ERK and P38 were detected by Western blot. Results:The incubation with carvacrol resulted in a significant inhibition of BGC-823 cell proliferation. After the treatment with carvacrol,the apoptotic rate was significantly increased( 0 μmol · L-1 vs 10 μmol · L-1 ,P<0. 000 1;0 μmol·L-1 vs 20 μmol·L-1 ,P<0. 000 1;0 μmol·L-1 vs 40 μmol·L-1 ,P<0. 000 1;0 μmol·L-1 vs 80μmol·L-1 ,P<0. 000 1)and the invasion ability was decreased(0 μmol·L-1 vs 80 μmol·L-1 ,P<0. 000 1). The expression of caspase-9(0 μmol·L-1 vs 80 μmol·L-1,P<0. 000 1)and TIMP-1 was increased(P<0. 000 1),PRAP fragment occurred(P<0. 000 1)and P38 signaling pathway was activated in the carvacrol treated group,while the expression of MMP-9 activity of ERK signa-ling pathway was inhibited(P<0.000 1). Conclusion:Carvacrol can inhibit cell growth and invasion,and induce cell apoptosis, which is closely related to MAPK signaling pathway.