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 investigate the hepatitis B surface antigen (HBsAg) clearance condition and its predictive factors after treatment with nucleos(t)ide analogues to pegylated interferon-α add-on therapy in patients with chronic hepatitis B.Methods:Patients with chronic hepatitis B who visited the First Affiliated Hospital of Zhengzhou University from 2018~2019 were prospectively enrolled. HBsAg≤ 1500 IU/mL, hepatitis B e antigen-negative, HBV DNA undetectable, received antiviral treatment with nucleos(t)ide analogues for at least one year, and pegylated interferon-α add-on therapy for 48 weeks were included. The primary endpoint of study was to determine the proportion of HBsAg clearance at 72 weeks. Concurrently, the predictive factors for HBsAg clearance were analyzed. Quantitative and qualitative data were analyzed using a t-test or non-parametric test and a Fisher's exact test.Results:A total of 38 cases were included in this study, of which 13 cases obtained HBsAg clearance at 48 weeks of therapy and another six cases obtained HBsAg clearance throughout the extended treatment period of 72 weeks, accounting for 50.00% of all enrolled patients. There was a significant difference in HBsAg dynamics between the HBsAg clearance group and the non-clearance group (P < 0.05). Univariate logistic regression analysis showed that patients' age, baseline, 12-and 24-week HBsAg levels, and early HBsAg reduction were predictive factors for HBsAg clearance at 72 weeks of treatment. Multivariate logistic regression analysis showed that age (OR = 1.311; P = 0.016; 95% confidence interval: 1.051~1.635) and HBsAg levels at 24 weeks of treatment (OR = 4.481; P = 0.004; 95% confidence interval: 1.634~12.290) were independent predictors for HBsAg clearance.Conclusion:Hepatitis B e antigen-negative, nucleos(t)ide analogue treated, HBsAg ≤ 1500 IU/mL, and HBV DNA undetectable, peg-IFNα add-on treatment for 48 weeks could promote HBsAg clearance in patients with chronic hepatitis B. Six of the sixteen cases (37.50%) who did not obtain HBsAg clearance at week 48 did so with the course of therapy extended to week 72. Hence, the optimal individualized treatment strategy should be customized according to the predictors rather than the fixed 48-week course. Age (≤ 38), baseline HBsAg level (≤2.86 log 10IU/ml), HBsAg level at 24 weeks (≤ 0.92 log 10IU/ml), and 12-week HBsAg decrease from baseline (≥ 0.67 log 10IU/ml) indicate that patients are highly likely to obtain HBsAg clearance at the 72 weeks of combination therapy, in which the combined indicator based on HBsAg level ≤0.92 log 10IU/ml at 24 weeks will identify 85.0% to 100.0% of patients with HBsAg clearance.

Objective To explore chromobox protein homolog 2 (CBX2) expressions in relation to clinical features of patients and elucidate its role in the progression of hepatocellular carcinoma.Methods Using the Cancer Genome Atlas (TCGA) database,R language was used to analyze the distribution of differentially expressed mRNA in hepatocellular carcinoma.The different expression of CBX2 in HCC and adjacent tissues and its relationship with survival and clinical characteristics of patients were further analyzed.The expression of CBX2 in liver tissues,liver cancer tissue,and L02,HepG2 and SMMC-7721 cell lines was detected by real time-PCR and western blot.The expression of CBX2 was interfered by siRNA in hepatoma cell line.MTT,colony formation,transwell assays,and flow cytometry were used to identify the proliferation,apoptosis,invasion and clone-formation ability of HepG2 and SMMC-7721 cells after CBX2 down-regulation.According to the different data,t-test,ANOVA,chi-square test,and COX regression model were used for statistical analysis.Survival curve was plotted through Kaplan-Meier method.Results TCGA public database analysis showed that the expression of CBX2 mRNA in hepatocellular carcinoma tissues (7.296 ± 1.6115) was significantly higher than normal liver tissues (4.706 ± 0.940) (P =0.000).In addition,the overall survival time of patients with low CBX2 mRNA expression was significantly longer than that of patients with high CBX2 mRNA expression [(5.971 ± 0.411) years vs.(4.650 ± 0.503) years,P =0.001].The expression level of CBX2 mRNA was correlated with the pathological TNM stage (P =0.025) and differentiation degree (P ＜ 0.001) of liver cancer.COX regression analysis showed that CBX2 mRNA expression was an independent predictor of patient survival (P =0.013).siRNA was transfected and compared with the blank control group.The transgenic ability of HepG2 and SMMC-77221 cells decreased significantly at 72h (P ＜ 0.05) and 96h (P ＜ 0.05),and the apoptosis rate (11.430％ ± 0.215％) was higher than blank control group (6.6 00％ ± 0.170％) (P =0.003).The number of invasive cells ((both P ＜ 0.05) and relative colony forming cells ((both P ＜ 0.001) were significantly decreased.In 20 cases of tissue samples,the expression of CBX2 protein (relative expression level 3.020 ±0.269) in liver cancer was higher than that in adjacent tissues (relative expression level 0.886±0.065) (P ＜ 0.001).The overall survival time of patients with low CBX2 expression in liver cancer was longer than that of patients with high expression [(3.670 + 0.576) years vs.(0.834 + 0.153) years,P =0.004].Conclusion An evident high expression of CBX2 is an independent poor prognostic factor in hepatoma.Down-regulation of CBX2 expression can inhibit the progression of liver cancer.Therefore,CBX2 may be a prognostic biomarker and a new target for HCC treatment.

Objective To analyze the clinical data of brucellosis,and to provide references for brucellosis therapy.Methods The patients definitely diagnosed brucellosis at the First Affiliated Hospital of Zhengzhou University from January 2013 to June 2016 were assessed,data of clinical features,laboratory examination,treatment and prognosis were analyzed.Results Of all 99 cases,the mean age was (46.7 ± 15.7) years old,83 cases had a history of closely contacted with sheep,2 cases with pig and 1 case with cattle.The occupational distribution of patients included 90 farmers,1 veterinarian,2 cooks,6 children and students.All patients had clinical manifestations such as fever,fatigue,and sweating.There were 18 patients with back and joint pain,13 cases had abnormal manifestation on magnetic resonance imaging (MRI).Blood culture was positive in 54 (71.05％,54/76) and serum test tube agglutination test was positive in 61 (98.39％,61/62).Eighty-one patients received doxycycline combined with rifampicin treatment,six months laters,all patients were cured.Conclusions Sheep are the main sources of infection for brucellosis.Fatigue,sweaty and fever are the most common symptoms,and osteoarticular is the most frequently involved.Serum agglutination test and blood culture are important tests for diagnosis of brucellosis.Doxycycline combined with rifampicin was the most common used antibiotics regimen.Early,combined,regular,full-course antibiotic treatment has a better prognosis.

Objective To investigate the clinical features of myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA) associated hypertrophic pachymeningitis (HP).Methods Clinical data of 4 casesdiagnosed with MPO-ANCA vasculitis complicated with HP in our hospital were analyzed retrospectively and the related literaturewere reviewed.Results Four male patients with an age range from 44 to 66 years were diagnosed with ANCA-associated HP.The main clinical manifestations included headache and withvarious degree ofmultiple cranial paralysis.During active phase of the disease,all patients showed perinuclear(p)-ANCA positive,elevated levels of inflammatory biomarkers and titers of MPO-ANCA,whereas renal function,cytoplasmic (c)-ANCA and protease 3 (PR3)-ANCA were negative.Contrast-enhanced cranial magnetic resonance imaging (MRI) scan showed obviously thickened dura mater and sinusitis or mass in paranasal sinus.Four patients were sensitive to glucocorticoid.Three patients had a relapse during glucocorticoid tapering and were undercontrol when the dosage of glucocorticoid was increased and immunosuppressive agents were added.Levels of inflammatory biomarkers,titers of MPO-ANCA and p-ANCA recovered to normal,and the dural thickness on MRI was reduced in the remission stage.Conclusion MPO-ANCA associated HP is a type of central nervous system involvement in ANCA associated vasculitis (AAV).It involves the upper respiratory tract more frequently,and less frequently progresses to systemic AAV.This should be taken into consideration when middle-aged and elderly patients presented with headache and multiple cranial neuropathies.Enhanced MRI is the preferred examination for diagnosis,and dural biopsy should be done when necessary.

In 2010,studies found that achievement of sustained virologic response after treatment in patients with chronic HCV infection meant cure;the overall cure rate of interferon combined with ribavirin around the world was about 60％,and the result of real-world study in China was 71.1％.In 2013,the US took the lead in launching the direct-acting antiviral agents (DAAs) for HCV,and the cure rate was increased to almost 100％.However,recent studies have found that the incidence of hepatocellular carcinoma (HCC) tends to increase after the application of DAAs,and there are still controversies over whether DAAs are the major cause of this phenomenon.In our opinion,this phenomenon is caused by the fact that the indications for DAAs cover more end-stage hepatitis C patients who are intrinsically the high-risk population of HCC;however,it cannot be excluded that DAAs may lead to the change in the body's antitumor immune status.Studies are needed in the future to examine this issue at the molecular level.

Objective To investigate the expression of PD-1 on CD4 +and CD8 + T cells in patients with HBeAg-positive chronic hepatitis B(CHB)treated with telbivudine.Methods Fifty-six HBeAg-positive CHB patients admitted in the First Affiliated Hospital of Zhengzhou University during January 201 3 and June 201 4 were enrolled in this study.The expression of PD-1 on CD4 +and CD8 + T cells was detected with flow cytometry at baseline,24,48 and 72 wks after telbivudine treatment.The relationship of PD-1 expression with alanine aminotransferase (ALT)level,HBeAg seroconversion and HBV DNA loads was analyzed.t test and completely random variance analysis were used to analyze the data.Results The PD-1 expression on CD4 + and CD8 + T cells at baseline was higher in patients with low ALT levels compared to those with high ALT levels(t =1 2.20 and 9.69,both P <0.01 ),while higher levels of PD-1 expression was also observed in patients with high HBV DNA load (≥5 lgIU /mL)compared to those with low HBV DNA load (t =4.39 and 4.85,both P <0.01 ).PD-1 levels on CD4 + and CD8 + T cells presented a declining trend after telbivudine treatment(F =6.98 and 8.97,both P <0.01 ),PD-1 expression in patients with HBeAg seroconversion showed lower levels compared with baseline values (t =1 8.45 and 1 8.01 ,both P <0.01 ). Conclusion In HBeAg-positive CHB patients,the expression of PD-1 on CD4 + and CD8 + T lymphocytes shows a decreasing trend during the treatment with telbivudine,indicating that antiviral therapy may alleviate the immunosuppression in these patients.

Objective To investigate the risk factors and predictive model for the occurrence of post-sustained virologic response (SVR)hepatocellular carcinoma in chronic hepatitis C (CHC)patients. Methods A total of 203 CHC patients hospitalized at the First Affiliated Hospital of Zhengzhou University from January 2006 to December 2014 who received antiviral therapy and achieved SVR were collected,including 11 post-SVR HCC cases.Risk factors for post-SVR HCC were estimated by Cox′s proportional hazards regression model.Cutoff value predicting risk of post-SVR HCC was determined by receiver operating characteristic curve.Results In Cox′s model,the risk of post-SVR HCC increased by 9.4-fold in patients with initial diagnosis as compensated cirrhosis compared to those with initial diagnosis as CHC.Increase in post-SVR albumin by per 1 g/L was associated with reduced risk by 20% for the occurrence of post-SVR HCC.Cut-off value of post-SVR albumin for the prediction of HCC was determined as ≤ 36.0 g/L with an area under the curve (AUC)of 0.809.A predictive model for post-SVR HCC was created based on initial diagnosis as compensated cirrhosis and post-SVR albumin ≤36.0 g/L with an AUC of 0.871 .The sensitivity,specificity and negative predictive value of the model were 0. 818,0.896 and 0.989,respectively.Conclusions Initial diagnosis as compensated cirrhosis combines with post-SVR albumin ≤36.0 g/L are risk factors for post-SVR HCC with ideal prediction value for the occurrence of post-SVR HCC in CHC patients.

Objective To investigate the dynamic changes of regulatory T cells (Treg ) and the surface expression of programmed death (PD)‐1 and the level of transforming growth factor (TGF )‐βduring antiviral treatment in patients with chronic hepatitis C (CHC) .Methods Eighty‐six CHC patients referred to the First Affiliated Hospital of Zhengzhou University from October 2012 to October 2013 were included ,and all of them were administered with pegylated interferon α‐2a and ribavirin .Thirty healthy controls were enrolled .The percentage of Treg cells ,PD‐1 expression and TGF‐β level were analyzed by flow cytometry at baseline and at time of achieving rapid virological response (RVR ) , early viral virological (EVR ) , end‐of‐treatment virological response (ETVR ) and sustained virological response (SVR) ,or not achieving SVR .Comparison between two groups was analyzed by t test .Results Among 86 CHC patients ,the proportions of RVR ,EVR ,ETVR ,and SVR at week 24 of follow‐up were 29 cases ,67 cases ,79 cases and 67 cases ,respectively .Percentage of Treg cells in CHC patients was much higher than that in healthy controls (10 .31 ± 5 .61 vs 2 .18 ± 0 .65 ,t = 2 .28 , P< 0 .05) .During antiviral therapy ,percentages of Treg cells declined ,not only in CHC patients with HCV genotype 1b (at baseline , RVR ,EVR ,and ETVR :14 .44 ± 3 .78 ,11 .01 ± 1 .79 ,8 .24 ± 2 .98 ,and 5 .36 ± 1 .47 ,respectively ) ,but also in those infected with HCV genotype 2a (at baseline ,RVR ,EVR ,and ETVR :12 .34 ± 2 .82 ,8 .99 ± 1 .68 ,7 .53 ± 2 .96 ,and 4 .79 ± 1 .23 ,respectively ) .Expressions of PD‐1 and TGF‐β also decreased .At baseline ,the expressions of PD‐1 in patients with SVR and without SVR were 29 .11 ± 14 .65 and 37 .73 ± 11 .65 ,respectively (t = 2 .15 , P = 0 .04) ,and the levels of TGF‐β were 41 .20 ± 18 .96 and 56 .75 ± 14 .42 ,respectively (t= 2 .66 ,P< 0 .01) .At week 24 ,the expressions of PD‐1 in patients with SVR and without SVR were 10 .36 ± 4 .81 and 36 .46 ± 10 .52 ,respectively (t= 13 .95 ,P< 0 .01) ,and the levels of TGF‐β were 10 .06 ± 4 .64 and 45 .23 ± 17 .85 , respectively ( t = 11 .85 , P < 0 .01 ) . Conclusions Percentages of Treg cells and expressions of PD‐1 and TGF‐β decrease during antiviral treatment in CHC patients .Thus ,it could be of assist to predict the treatment response by monitoring these parameters .

OBJECTIVETo study the CPS-II mechanism underlying the pathological process of elevated blood ammonia leading to liver injury.

METHODSAn in vitro hyperammonemia hepatocyte cell model was constructed by exposure to various concentrations of NH4Cl. The subsequent changes to cellular morphology were observed by microscopy. to cell apoptosis were determined by flow cytometry, and to mRNA and protein expression of CPS-II were examined by real-time PCR and western blotting, respectively.

RESULTSExposure to NH₄Cl led to dose-dependent morphological damage, apoptosis and necrosis of the hepatocytes. The apoptosis rate was significantly higher for the high-dose group than for the control (no exposure) group (24.7% ± 2.39% vs. 4.1% ± 0.78%, q =8.06, P less than 0.05). Expression of the CPS-II mRNA was significantly elevated in response to NH₄Cl exposure (vs. the control group; F=191.881, P < 0.05).The CPS-II mRNA expression level increased with increasing NH₄Cl concentration (grey values: 1.040 ± 0.045, 1.641 ± 0.123, 2.285 ± 0.167 and 3.347 ± 0.124, respectively). The CPS-II protein expression level was also significantly enhanced in response to the NH₄Cl exposures (CPS-II protein and internal GAPDH grey value ratios: 0.099 ± 0.0130, 0.143 ± 0.025, 0.161 ± 0.036 and 0.223 ± 0.042, respectively; t=3.825, 3.968 and 6.908, P less than 0.05).

CONCLUSIONCPS-II mRNA and protein expression levels become elevated with increase in the NH₄Cl concentrations, suggesting that in addition to the urea cycle, CPS-II may play an important role in the ammonia metabolism under the condition of hyperammonemia.