Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)enters host cells via the angiotensin-converting enzyme 2(ACE2)receptor.Mounting evidence has indicated the presence of hepatic SARS-CoV-2 infection and liver...Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)enters host cells via the angiotensin-converting enzyme 2(ACE2)receptor.Mounting evidence has indicated the presence of hepatic SARS-CoV-2 infection and liver injury in pa-tients with coronavirus disease 2019(COVID-19).Understanding the mechanisms of hepatic SARS-CoV-2 infection is crucial for addressing COVID-19–related liver pathology and developing targeted therapies.This editorial discusses the signi-ficance of ACE2 in hepatic SARS-CoV-2 infection,drawing on the research by Jacobs et al.Their findings indicate that hepatic ACE2 expression,frequency of hepatic SARS-CoV-2 infection,and severity of liver injury are elevated in patients with pre-existing chronic liver diseases.These data suggest that hepatic ACE2 could be a promising therapeutic target for COVID-19.展开更多
AIM: To study Hepatitis B virus (HBV) infection and its association with hepatocellular carcinoma (HCC) at the miRNA level.METHODS: Three cellular models were used to investigate miRNA expression changes during HBV in...AIM: To study Hepatitis B virus (HBV) infection and its association with hepatocellular carcinoma (HCC) at the miRNA level.METHODS: Three cellular models were used to investigate miRNA expression changes during HBV infection: human HepG2 hepatoblastoma cell line as a model without HBV infection;HepG2 cell line transfected with a 1.3-fold full-length HBV genome as an acute infection model;and HepG2.2.15 cell line,which is derived from HepG2 and stably transfected with a complete HBV genome,as a chronic infection model.The miRNA levels were examined using microarray technology.To explore the relationship between HBV infection and HCC genesis at the miRNA level,we downloaded from national center for biotechnology information Gene Expression Omnibus an miRNA expression dataset derived from HCC patients,most of whom are HBV carriers.We compared the miRNA expression alterations during HBV infection with those in HCC patients,by analyzing miRNA expression change profiles statistically.RESULTS: Seventy-seven and 48 miRNAs were differentially expressed during acute and chronic HBV infection,respectively.Among these miRNAs,25 were in common,the intersection of which was significant under the hypergeometric test (P = 1.3 × 10-11).Fourteen miRNAs were observed to change coherently in the acute and chronic infections,with one upregulated and 13 downregulated.Eleven showed inverse changes during the two phases of infection;downregulated in the acute infection and upregulated in the chronic infection.The results imply that common and specific mechanisms exist at the miRNA level during acute and chronic HBV infection.Besides,comparative analysis of the miRNA expression changes during HBV infection with those in HCC indicates that,although miRNA expression changes during HBV infection are distinct from those in HCC patients (P < 2.2 × 10-16),they exhibited significant correlations (P = 0.0229 for acute infection;P = 0.0084 for chronic infection).Perturbation of miRNA expression during chronic HBV infection was closer to that in HCC patients than that during acute HBV infection.This observation implies the contribution of miRNAs to HCC genesis from HBV infection.According to their patterns of differential expression in acute and chronic HBV infection,as well as in HCC,miRNAs of potential research interest could be identified,such as miR-18a/miR-18b,miR-106a,miR-221 and miR-101.For instance,the gradient expression alteration of miR-221 in the above three phases,which is downregulated in acute HBV infection,normally expressed in chronic HBV infection,and upregulated in HCC,indicates that it may be a key effector for progression of the disease.CONCLUSION: Our analysis provides insights into HBV infection and related HCC in relation to miRNAs,and reveals some candidate miRNAs for future studies.展开更多
AIM： To investigate whether the recombinant adenovirus induces the TNF-α-mediated apoptosis in vivo. METHODS： Human hepatocarcinoma cell line （HepG2） cells were transfected into BALB/c nude mice, and the tumor gr...AIM： To investigate whether the recombinant adenovirus induces the TNF-α-mediated apoptosis in vivo. METHODS： Human hepatocarcinoma cell line （HepG2） cells were transfected into BALB/c nude mice, and the tumor growth curve was drawn. We analyzed apoptosis in HepG2 cells by TUNEL, HE staining and electron microscopy. RESULTS： AdIκBαM was expressed stably and efficiently in HepG2 and could not be degraded by induction of TNF-α. Tumor growth in mice could be reduced remarkably if treated by AdIκBαM plus TNF-α. There was apoptosis of 〉 70% of cells treated with AdIκBαM plus TNF-α and about 50% of cells treated with AdIκBαM. In contrast, there was few cell apoptosis in HepG2 cells treated with phosphate buffered saline and AdIκBαM. HepG2 cells in mice also exhibited a high level of apoptosis after in vivo injection with AdIκBαM. The tumor growth curve indicated the tumor transfected with AdIκBαM could be restrained. CONCLUSION： AdIκBαM gene therapy greatly enhances apoptosis due to inhibition of an NF-κB-mediated antiapoptosis signaling pathway.展开更多
AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,t...AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,the levels of hepatitis B surface antigen(HBs Ag),hepatitis B e antigen(HBe Ag),and hepatitis B core antigen(HBc Ag) in the supernatants or cytoplasm were examined using by enzyme-linked immunosorbent assay and Western blot.The HBV pregenome RNA(pg RNA) was detected using reverse transcription-PCR and realtime fluorescent quantitative PCR(RT-q PCR),and viral DNA was detected using Southern blot and RT-q PCR.Cell proliferation after drug treatment was detected using the Brd U incorporation test and the trypan blue exclusion assay.Cell cycle and cell apoptosis were examined using flow cytometry and Western blot.RESULTS:Vincristine up-regulated HBV replication directly in vitro in a dose-dependent manner,and 24-h exposure to 0.1 μmol/L vincristine induced more than 4-fold and 3-fold increases in intracellular HBV DNA and the secretion of viral DNA,respectively.The expression of HBV pg RNA,intracellular HBs Ag and HBc Ag,and the secretion of HBe Ag were also increased significantly after drug treatment.Most importantly,vincristine promoted the cell excretion of HBV nucleocapsids instead of HBV Dane particles,and the nucleocapsids are closely related to the HBV pathogenesis.Furthermore,vincristine inhibited the proliferation of cells stably expressing HBV.The higher the concentration of the drug,the more significant the inhibition of the cell proliferation and the stronger the HBV replication ability in cells.Flow cytometry indicated that cell cycle arrest at S-phase was responsible for the cell proliferation inhibition.CONCLUSION:Vincristine has a strong stimulatory effect on HBV replication and induces cell cycle arrest,and cell proliferation inhibition may be conducive to viral replication.展开更多
AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×...AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×1012copies of linear HBV DNA/1×107 PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System.Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group.RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive≥2.1) respectively. HBeAg was negative (＜2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group.CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species.HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication.展开更多
OBJECTIVE: To clarify the natural history, of chronic hepatitis B so as to evaluate its long-term therapeutic outcome of the patients and the efficacy of antiviral drugs. METHODS: A cohort of 183 biopsy-proven chronic...OBJECTIVE: To clarify the natural history, of chronic hepatitis B so as to evaluate its long-term therapeutic outcome of the patients and the efficacy of antiviral drugs. METHODS: A cohort of 183 biopsy-proven chronic hepatitis B patients (mean age of 31.75±8.03 years, male/female ratio: 152:31) and 247 controls were followed up retrospectively for 11.81±4.08 years. This study was focused on long-term clinical outcome including the rates of liver cirrhosis, hepatocellular carcinoma and death, apart from the long-term effect of antiviral drugs and prognostic factors. RESULTS: In the 183 chronic hepatitis B patients, 22 (12.02%) developed liver cirrhosis, 12 (6.56%) developed hepatocellular carcinoma, and 20 (10.93%) died. The 5-, 10- and 15-year survival rates were 97. 27%, 91.62%, and 84.47%, respectively. The 5-, 10- and 15-year incidence rates of HCC were O, 3.19%, and 11.56%, respectively. In the 247 controls, 6 (2.43%) died; none of them developed cirrhosis or HCC. The rates of death, liver cirrhosis, and HCC in the hepatitis B patients were markedly different (P<0. 005) compared with the controls. The overall mortality of hepatitis B patients was 4.5-fold higher than the general population. Cox multiple regression analysis showed that old age, severe histological injury, and positive HBeAg were closely related to liver cirrhosis; old age, severe histological injury, and male were major factors leading to death. The independent variable of predicted HCC was not found. CONCLUSION: The long-term outcome of hepatitis B patients is poor and the efficacy of antiviral drugs needs further study.展开更多
The achievement of a functional cure for chronic hepatitis B(CHB)remains limited to a minority of patients treated with currently approved drugs.The primary objective in developing new anti-HBV drugs is to enhance the...The achievement of a functional cure for chronic hepatitis B(CHB)remains limited to a minority of patients treated with currently approved drugs.The primary objective in developing new anti-HBV drugs is to enhance the functional cure rates for CHB.A critical prerequisite for the functional cure of CHB is a substantial reduction,or even eradication of covalently closed circular DNA(cccDNA).Within this context,the changes in cccDNA levels during treatment become as a pivotal concern.We have previously analyzed the factors influencing cccDNA dynamics and introduced a preliminary classification of hepatitis B treatment strategies based on these dynamics.In this review,we employ a systems thinking perspective to elucidate the fundamental aspects of the HBV replication cycle and to rationalize the classification of treatment strategies according to their impact on the dynamic equilibrium of cccDNA.Building upon this foundation,we categorize current anti-HBV strategies into two distinct groups and advocate for their combined use to significantly reduce cccDNA levels within a well-defined timeframe.展开更多
The landscape of hepatitis B virus(HBV)integration in the plasma cell-free DNA(cfDNA)of HBV-infected patients with different stages of liver diseases[chronic hepatitis B(CHB),liver cirrhosis(LC),and hepatocellular car...The landscape of hepatitis B virus(HBV)integration in the plasma cell-free DNA(cfDNA)of HBV-infected patients with different stages of liver diseases[chronic hepatitis B(CHB),liver cirrhosis(LC),and hepatocellular carcinoma(HCC)]remains unclear.In this study,we developed an improved strategy for detecting HBV DNA integration in plasma cfDNA,based on DNA probe capture and next-generation sequencing.Using this optimized strategy,we successfully detected HBV integration events in chimeric artificial DNA samples and HBV-infected HepG2-NTCP cells at day one post infection,with high sensitivity and accuracy.The characteristics of HBV integration events in the HBV-infected HepG2-NTCP cells and plasma cfDNA from HBV-infected individuals(CHB,LC,and HCC)were further investigated.A total of 112 and 333 integration breakpoints were detected in the HepG2-NTCP cells and 22 out of 25(88%)clinical HBV-infected samples,respectively.In vivo analysis showed that the normalized number of support unique sequences(nnsus)in HCC was significantly higher than in CHB or LC patients(P values<0.05).All integration breakpoints are randomly distributed on human chromosomes and are enriched in the HBV genome around nt 1800.The majority of integration breakpoints(61.86%)are located in the gene-coding region.Both non-homologous end-joining(NHEJ)and microhomology-mediated end-joining(MMEJ)interactions occurred during HBV integration across the three different stages of liver diseases.Our study provides evidence that HBV DNA integration can be detected in the plasma cfDNA of HBV-infected patients,including those with CHB,LC,or HCC,using this optimized strategy.展开更多
Dear Editor,A common reason for drug failure during long-term treatment of chronic hepatitis B with nucleot(s)ide analogues(NUCs)is the emergence of drug resistance(Das et al.,2001).Most primary NUCs-resistant mutatio...Dear Editor,A common reason for drug failure during long-term treatment of chronic hepatitis B with nucleot(s)ide analogues(NUCs)is the emergence of drug resistance(Das et al.,2001).Most primary NUCs-resistant mutations identified in clinical samples have been limited to a minority of amino acids(usually less展开更多
On 24 November,a new detected variant B.1.1.529 of SARS-CoV-2 by South Africa was reported to WHO.After only 2 days,this variant was designated as“variant of concern”(VOC)and named as Omicron.In the past few weeks,O...On 24 November,a new detected variant B.1.1.529 of SARS-CoV-2 by South Africa was reported to WHO.After only 2 days,this variant was designated as“variant of concern”(VOC)and named as Omicron.In the past few weeks,Omicron had reported from more than 80 countries.It has been reported as the dominant SARS-CoV-2 in U.S.due to the rapid spread of Omicron.A new wave of infection driven by Omicron is in progress.展开更多
Coronaviruses are enveloped,positive-stranded RNA viruses that contain the largest known RNA genomes to date.As severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continues to circulate in the human population...Coronaviruses are enveloped,positive-stranded RNA viruses that contain the largest known RNA genomes to date.As severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continues to circulate in the human population,multiple mutations have accumulated over time,which may affect its transmission,virulence and antigenicity.展开更多
Many countries around the world have seen a sharp rise in COVID-19 cases since the beginning of October due to the second wave of the pandemic.A decline in the antibody response to severe acute respiratory syndrome co...Many countries around the world have seen a sharp rise in COVID-19 cases since the beginning of October due to the second wave of the pandemic.A decline in the antibody response to severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which was reported exclusively in the early month,increases the risk of reinfection for convalescent individuals.There is a current need to follow the maintenance of specific antibodies against SARS-CoV-2.展开更多
Dear Editor,Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is a major public health issue.The spike(S)protein mutation D614G became dominant in SARS-CoV-2 during...Dear Editor,Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is a major public health issue.The spike(S)protein mutation D614G became dominant in SARS-CoV-2 during a global pandemic,which displayed increased infectivity.1 Entry of a virus into host cells is one of the most critical steps in the viral life cycle.Since blockade of the entry process is a promising therapeutic option for COVID-19,research attention has been focused on the discovery of viral entry inhibitors.Although SARS-CoV-2 entry inhibitor development is very attractive,no candidates have progressed into clinical trials yet.展开更多
Apolipoprotein E(APOE)plays a pivotal role in lipid including cholesterol metabolism.The APOE 4(APOE4)allele is a major genetic risk factor for Alzheimer's and cardiovascular diseases.Although APOE has recently be...Apolipoprotein E(APOE)plays a pivotal role in lipid including cholesterol metabolism.The APOE 4(APOE4)allele is a major genetic risk factor for Alzheimer's and cardiovascular diseases.Although APOE has recently been associated with increased susceptibility to infections of several viruses,whether and how APOEand its isoforms affect SARS-CoV-2 infection remains unclear.Here,we show that serum concentrations of APOE correlate inversely with levels of cytokine/chemokine in 73 COVID-19 patients.Utilizing multiple protein interaction assays,we demonstrate that APOE3 and APOE4 interact with the SARS-CoV-2 receptor ACE2;and APOE/ACE2 interactions require zinc metallopeptidase domain of ACE2,a key docking site for SARS-CoV-2 Spike protein.In addition,immuno-imaging assays using confocal,super-resolution,and transmission electron microscopies reveal that both APOE3 and APOE4 reduce ACE2/Spikemediated viral entry into cells.Interestingly,while having a comparable binding affinity to ACE2,APOE4 inhibits viral entry to a lesser extent compared to APOE3,which is likely due to APOE4's more compact structure and smaller spatial obstacle to compete against Spike binding to ACE2.Furthermore,APOE e4 carriers clinically correlate with increased SARS-CoV-2 infection and elevated serum inflammatory factors in 142 COVID-19 patients assessed.Our study suggests a regulatory mechanism underlying SARS-CoV-2 infection through APOE interactions with ACE2,which may explain in part increased COVID-19 infection and disease severity in APOE e4 carriers.展开更多
In December 2019,the corona virus disease 2019(COVID-19)caused by novel coronavirus(SARS-CoV-2)emerged in Wuhan,China and rapidly spread worldwide.Few information on clinical features and immunological profile of COVI...In December 2019,the corona virus disease 2019(COVID-19)caused by novel coronavirus(SARS-CoV-2)emerged in Wuhan,China and rapidly spread worldwide.Few information on clinical features and immunological profile of COVID-19 in paediatrics.The clinical features and treatment outcomes of twelve paediatric patients confirmed as COVID-19 were analyzed.The immunological features of children patients was investigated and compared with twenty adult patients.The median age was 14.5-years(range from 0.64 to 17),and six of the patients were male.The average incubation period was 8 days.Clinically,cough(9/12,75%)and fever(7/12,58.3%)were the most common symptoms.Four patients(33.3%)had diarrhea during the disease.As to the immune profile,children had higher amount of total T cell,CD8t T cell and B cell but lower CRP levels than adults(P<0.05).Ground-glass opacity(GGO)and local patchy shadowing were the typical radiological findings on chest CT scan.All patients received antiviral and symptomatic treatment and the symptom relieved in 3e4 days after admitted to hospital.The paediatric patients showed mild symptom but with longer incubation period.Children infected with SARS-CoV-2 had different immune profile with higher T cell amount and low inflammatory factors level,which might ascribed to the mild clinical symptom.We advise that nucleic acid test or examination of serum IgM/IgG antibodies against SARS-CoV-2 should be taken for children with exposure history regardless of clinical symptom.展开更多
Coronavirus disease 2019(COVID-19)is caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The Spike protein that mediates coronavirus entry into host cells is a major target for COVID-19 vaccines and ...Coronavirus disease 2019(COVID-19)is caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The Spike protein that mediates coronavirus entry into host cells is a major target for COVID-19 vaccines and antibody therapeutics.However,multiple variants of SARS-CoV-2 have emerged,which may potentially compromise vaccine effectiveness.Using a pseudovirus-based assay,we evaluated SARS-CoV-2 cell entry mediated by the viral Spike B.1.617 and B.1.1.7 variants.We also compared the neutralization ability of monoclonal antibodies from convalescent sera and neutralizing antibodies(NAbs)elicited by CoronaVac(inactivated vaccine)and ZF2001(RBD-subunit vaccine)against B.1.617 and B.1.1.7 variants.Our results showed that,compared to D614G and B.1.1.7 variants,B.1.617 shows enhanced viral entry and membrane fusion,as well as more resistant to antibody neutralization.These findings have important implications for understanding viral infectivity and for immunization policy against SARS-CoV-2 variants.展开更多
Correction to:Cellular&Molecular Immunology https:/doi.org/10.1038/s41423-020-00605-4,published online 08 January 2021 The licence information was missing from this article and should havebeen CC-BY.The original a...Correction to:Cellular&Molecular Immunology https:/doi.org/10.1038/s41423-020-00605-4,published online 08 January 2021 The licence information was missing from this article and should havebeen CC-BY.The original article has been corrected.展开更多
基金Supported by National Natural Science Foundation of China,No.82172915,No.81972648,and No.81773011Chongqing Medical University Program for Youth Innovation in Future Medicine,No.W0084+1 种基金Science and Technology Innovation Project of Chongqing Medical Universityand Chongqing Postdoctoral Science Foundation,No.CSTB2023NSCQ-BHX0134.
文摘Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)enters host cells via the angiotensin-converting enzyme 2(ACE2)receptor.Mounting evidence has indicated the presence of hepatic SARS-CoV-2 infection and liver injury in pa-tients with coronavirus disease 2019(COVID-19).Understanding the mechanisms of hepatic SARS-CoV-2 infection is crucial for addressing COVID-19–related liver pathology and developing targeted therapies.This editorial discusses the signi-ficance of ACE2 in hepatic SARS-CoV-2 infection,drawing on the research by Jacobs et al.Their findings indicate that hepatic ACE2 expression,frequency of hepatic SARS-CoV-2 infection,and severity of liver injury are elevated in patients with pre-existing chronic liver diseases.These data suggest that hepatic ACE2 could be a promising therapeutic target for COVID-19.
基金Supported by Ministry of Science and Technology of China,No.2007CB516810National Natural Science Foundation of China,Nos.30800971 and 30900270China Postdoctoral Science Foundation,No.20070420731
文摘AIM: To study Hepatitis B virus (HBV) infection and its association with hepatocellular carcinoma (HCC) at the miRNA level.METHODS: Three cellular models were used to investigate miRNA expression changes during HBV infection: human HepG2 hepatoblastoma cell line as a model without HBV infection;HepG2 cell line transfected with a 1.3-fold full-length HBV genome as an acute infection model;and HepG2.2.15 cell line,which is derived from HepG2 and stably transfected with a complete HBV genome,as a chronic infection model.The miRNA levels were examined using microarray technology.To explore the relationship between HBV infection and HCC genesis at the miRNA level,we downloaded from national center for biotechnology information Gene Expression Omnibus an miRNA expression dataset derived from HCC patients,most of whom are HBV carriers.We compared the miRNA expression alterations during HBV infection with those in HCC patients,by analyzing miRNA expression change profiles statistically.RESULTS: Seventy-seven and 48 miRNAs were differentially expressed during acute and chronic HBV infection,respectively.Among these miRNAs,25 were in common,the intersection of which was significant under the hypergeometric test (P = 1.3 × 10-11).Fourteen miRNAs were observed to change coherently in the acute and chronic infections,with one upregulated and 13 downregulated.Eleven showed inverse changes during the two phases of infection;downregulated in the acute infection and upregulated in the chronic infection.The results imply that common and specific mechanisms exist at the miRNA level during acute and chronic HBV infection.Besides,comparative analysis of the miRNA expression changes during HBV infection with those in HCC indicates that,although miRNA expression changes during HBV infection are distinct from those in HCC patients (P < 2.2 × 10-16),they exhibited significant correlations (P = 0.0229 for acute infection;P = 0.0084 for chronic infection).Perturbation of miRNA expression during chronic HBV infection was closer to that in HCC patients than that during acute HBV infection.This observation implies the contribution of miRNAs to HCC genesis from HBV infection.According to their patterns of differential expression in acute and chronic HBV infection,as well as in HCC,miRNAs of potential research interest could be identified,such as miR-18a/miR-18b,miR-106a,miR-221 and miR-101.For instance,the gradient expression alteration of miR-221 in the above three phases,which is downregulated in acute HBV infection,normally expressed in chronic HBV infection,and upregulated in HCC,indicates that it may be a key effector for progression of the disease.CONCLUSION: Our analysis provides insights into HBV infection and related HCC in relation to miRNAs,and reveals some candidate miRNAs for future studies.
基金Supported by the Foundation of Hi-tech Research and Develop-ment Program of China (863 Program), No. 2001AA217121Co-fi rst-author: Li-Ping Jia
文摘AIM： To investigate whether the recombinant adenovirus induces the TNF-α-mediated apoptosis in vivo. METHODS： Human hepatocarcinoma cell line （HepG2） cells were transfected into BALB/c nude mice, and the tumor growth curve was drawn. We analyzed apoptosis in HepG2 cells by TUNEL, HE staining and electron microscopy. RESULTS： AdIκBαM was expressed stably and efficiently in HepG2 and could not be degraded by induction of TNF-α. Tumor growth in mice could be reduced remarkably if treated by AdIκBαM plus TNF-α. There was apoptosis of 〉 70% of cells treated with AdIκBαM plus TNF-α and about 50% of cells treated with AdIκBαM. In contrast, there was few cell apoptosis in HepG2 cells treated with phosphate buffered saline and AdIκBαM. HepG2 cells in mice also exhibited a high level of apoptosis after in vivo injection with AdIκBαM. The tumor growth curve indicated the tumor transfected with AdIκBαM could be restrained. CONCLUSION： AdIκBαM gene therapy greatly enhances apoptosis due to inhibition of an NF-κB-mediated antiapoptosis signaling pathway.
基金Supported by National Natural Science Foundation of China,No.81201282 and No.30901280Grants from Chongqing Natural Science Foundation,No.cstc2012jj A10047the PhD Programs Foundation of the Ministry of Education of China,No.20125503120004
文摘AIM:To observe the effect of vincristine on hepatitis B virus(HBV) replication in vitro and to study its possible mechanisms.METHODS:Vincristine was added to the cultures of two cell lines stably expressing HBV.Then,the levels of hepatitis B surface antigen(HBs Ag),hepatitis B e antigen(HBe Ag),and hepatitis B core antigen(HBc Ag) in the supernatants or cytoplasm were examined using by enzyme-linked immunosorbent assay and Western blot.The HBV pregenome RNA(pg RNA) was detected using reverse transcription-PCR and realtime fluorescent quantitative PCR(RT-q PCR),and viral DNA was detected using Southern blot and RT-q PCR.Cell proliferation after drug treatment was detected using the Brd U incorporation test and the trypan blue exclusion assay.Cell cycle and cell apoptosis were examined using flow cytometry and Western blot.RESULTS:Vincristine up-regulated HBV replication directly in vitro in a dose-dependent manner,and 24-h exposure to 0.1 μmol/L vincristine induced more than 4-fold and 3-fold increases in intracellular HBV DNA and the secretion of viral DNA,respectively.The expression of HBV pg RNA,intracellular HBs Ag and HBc Ag,and the secretion of HBe Ag were also increased significantly after drug treatment.Most importantly,vincristine promoted the cell excretion of HBV nucleocapsids instead of HBV Dane particles,and the nucleocapsids are closely related to the HBV pathogenesis.Furthermore,vincristine inhibited the proliferation of cells stably expressing HBV.The higher the concentration of the drug,the more significant the inhibition of the cell proliferation and the stronger the HBV replication ability in cells.Flow cytometry indicated that cell cycle arrest at S-phase was responsible for the cell proliferation inhibition.CONCLUSION:Vincristine has a strong stimulatory effect on HBV replication and induces cell cycle arrest,and cell proliferation inhibition may be conducive to viral replication.
基金Supported by the National Natural Science Foundation of China,No. 39670340the Applied Basic Research Programs of ScienceTechnology Commission Foundation of Chongqing, No.20021889
文摘AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×1012copies of linear HBV DNA/1×107 PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System.Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group.RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive≥2.1) respectively. HBeAg was negative (＜2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group.CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species.HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication.
基金The project was supported by the fund for young hepatologists,Chongqing,China(Chinese Medical Association 1998).
文摘OBJECTIVE: To clarify the natural history, of chronic hepatitis B so as to evaluate its long-term therapeutic outcome of the patients and the efficacy of antiviral drugs. METHODS: A cohort of 183 biopsy-proven chronic hepatitis B patients (mean age of 31.75±8.03 years, male/female ratio: 152:31) and 247 controls were followed up retrospectively for 11.81±4.08 years. This study was focused on long-term clinical outcome including the rates of liver cirrhosis, hepatocellular carcinoma and death, apart from the long-term effect of antiviral drugs and prognostic factors. RESULTS: In the 183 chronic hepatitis B patients, 22 (12.02%) developed liver cirrhosis, 12 (6.56%) developed hepatocellular carcinoma, and 20 (10.93%) died. The 5-, 10- and 15-year survival rates were 97. 27%, 91.62%, and 84.47%, respectively. The 5-, 10- and 15-year incidence rates of HCC were O, 3.19%, and 11.56%, respectively. In the 247 controls, 6 (2.43%) died; none of them developed cirrhosis or HCC. The rates of death, liver cirrhosis, and HCC in the hepatitis B patients were markedly different (P<0. 005) compared with the controls. The overall mortality of hepatitis B patients was 4.5-fold higher than the general population. Cox multiple regression analysis showed that old age, severe histological injury, and positive HBeAg were closely related to liver cirrhosis; old age, severe histological injury, and male were major factors leading to death. The independent variable of predicted HCC was not found. CONCLUSION: The long-term outcome of hepatitis B patients is poor and the efficacy of antiviral drugs needs further study.
基金supported by grants from the National Key R&D Program of China(2022YFA1303600)the Natural Science Foundation of Chongqing(cstc2021jcyj-msxmX0298)+2 种基金the Science and Technology Research Program of Chongqing Municipal Education Commission(Grant No.KJZD-K202200409)the 111 Project(No.D20028),Key Laboratory of Molecular Biology on Infectious Diseases,Ministry of Education,Chongqing Medical University(No.202104)CQMU Program for Youth Innovation in Future Medicine(No.W0049).
文摘The achievement of a functional cure for chronic hepatitis B(CHB)remains limited to a minority of patients treated with currently approved drugs.The primary objective in developing new anti-HBV drugs is to enhance the functional cure rates for CHB.A critical prerequisite for the functional cure of CHB is a substantial reduction,or even eradication of covalently closed circular DNA(cccDNA).Within this context,the changes in cccDNA levels during treatment become as a pivotal concern.We have previously analyzed the factors influencing cccDNA dynamics and introduced a preliminary classification of hepatitis B treatment strategies based on these dynamics.In this review,we employ a systems thinking perspective to elucidate the fundamental aspects of the HBV replication cycle and to rationalize the classification of treatment strategies according to their impact on the dynamic equilibrium of cccDNA.Building upon this foundation,we categorize current anti-HBV strategies into two distinct groups and advocate for their combined use to significantly reduce cccDNA levels within a well-defined timeframe.
基金supported by the National Key Research and Development Program of China(2018YFE0107500)the CQMU Program for Youth Innovation in Future Medicine(W0160)to Yuan Hu+3 种基金Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJQN202100422)Scientific Research Projects in Yuzhong District of Chongqing(20210116)to HaiJun Dengthe 111 Project(D20028)to AiLong Huangand the Natural Science Foundation of Chongqing(cstc2021jcyjmsxmX0202)to Hongchun Luo.
文摘The landscape of hepatitis B virus(HBV)integration in the plasma cell-free DNA(cfDNA)of HBV-infected patients with different stages of liver diseases[chronic hepatitis B(CHB),liver cirrhosis(LC),and hepatocellular carcinoma(HCC)]remains unclear.In this study,we developed an improved strategy for detecting HBV DNA integration in plasma cfDNA,based on DNA probe capture and next-generation sequencing.Using this optimized strategy,we successfully detected HBV integration events in chimeric artificial DNA samples and HBV-infected HepG2-NTCP cells at day one post infection,with high sensitivity and accuracy.The characteristics of HBV integration events in the HBV-infected HepG2-NTCP cells and plasma cfDNA from HBV-infected individuals(CHB,LC,and HCC)were further investigated.A total of 112 and 333 integration breakpoints were detected in the HepG2-NTCP cells and 22 out of 25(88%)clinical HBV-infected samples,respectively.In vivo analysis showed that the normalized number of support unique sequences(nnsus)in HCC was significantly higher than in CHB or LC patients(P values<0.05).All integration breakpoints are randomly distributed on human chromosomes and are enriched in the HBV genome around nt 1800.The majority of integration breakpoints(61.86%)are located in the gene-coding region.Both non-homologous end-joining(NHEJ)and microhomology-mediated end-joining(MMEJ)interactions occurred during HBV integration across the three different stages of liver diseases.Our study provides evidence that HBV DNA integration can be detected in the plasma cfDNA of HBV-infected patients,including those with CHB,LC,or HCC,using this optimized strategy.
基金supported in part by grants from the National Natural Science Foundation of China(81671997)Chongqing Science&Technology Commission(cstc2015jcyj A10023)+1 种基金Chongqing Education Commission(CYB15099)the Program for Innovation Team of Higher Education in Chongqing(CXTDX 201601015)
文摘Dear Editor,A common reason for drug failure during long-term treatment of chronic hepatitis B with nucleot(s)ide analogues(NUCs)is the emergence of drug resistance(Das et al.,2001).Most primary NUCs-resistant mutations identified in clinical samples have been limited to a minority of amino acids(usually less
基金funding support from the China National Natural Science Foundation(grant no.U20A20392)the 111 Project(No.D20028),the Research Fund Program of the Key Laboratory of Molecular Biology for Infectious Diseases,CQMU(No.202105,202102)+6 种基金the Emergency Project from the Science&Technology Commission of Chongqing(cstc2020jscx-fyzx0053)the Emergency Project for Novel Coronavirus Pneumonia from the Chongqing Medical University(CQMUNCP0302)the Leading Talent Program of CQ CSTC(CSTCCXLJRC201719)a Major National Science&Technology Program grant(2017ZX10202203)the Science&Technology Commission of China,National Natural Science Foundation of China(Grant No.82102361)China Postdoctoral Science Foundation(2021M693924),Natural Science Foundation of Chongqing,China(cstc2021jcyj-bshX0115)Chongqing Postdoctoral Science Special Foundation(2010010005216630).
文摘On 24 November,a new detected variant B.1.1.529 of SARS-CoV-2 by South Africa was reported to WHO.After only 2 days,this variant was designated as“variant of concern”(VOC)and named as Omicron.In the past few weeks,Omicron had reported from more than 80 countries.It has been reported as the dominant SARS-CoV-2 in U.S.due to the rapid spread of Omicron.A new wave of infection driven by Omicron is in progress.
基金funding support from the Emergency Project from the Science&Technology Commission of Cho ngqing(cstc2020jscx-dxwtB0050,cstc2020jscx-fyzx0053)the Emergency Project for Novel Coronavirus Pneumonia from Chongqing Medical University(CQMUNCP0302)+2 种基金the Key Laboratory of Infectious Diseases(CQMU,202005)the Leading Talent Program of CQ CSTC(CSTCCXURC201719)a Major National Science&Technology Program grant(2017ZX10202203)from the Science&Technology Commission of China.
文摘Coronaviruses are enveloped,positive-stranded RNA viruses that contain the largest known RNA genomes to date.As severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continues to circulate in the human population,multiple mutations have accumulated over time,which may affect its transmission,virulence and antigenicity.
基金We acknowledge funding support from the Key Laboratory of Infectious Diseases(CQMU,202005)the Emergency Project from the Science&Technology Commission of Chongqing(cstc2020jscx-fyzx0053)+2 种基金the Emergency Project for Novel Coronavirus Pneumonia from Chongqing Medical University(CQMUNCP0302)the Leading Talent Program of CQ CSTC(CSTCCXLJRC201719)and a Major National Science&Technology Program grant(2017ZX10202203)from the Science&Technology Commission of China.
文摘Many countries around the world have seen a sharp rise in COVID-19 cases since the beginning of October due to the second wave of the pandemic.A decline in the antibody response to severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which was reported exclusively in the early month,increases the risk of reinfection for convalescent individuals.There is a current need to follow the maintenance of specific antibodies against SARS-CoV-2.
基金This work was supported by the Key Laboratory of Infectious Diseases,CQMU,202001the Science and Technology Research Program of Chongqing Municipal Education Commission(KJQN202000418 to L-Y.H.)+1 种基金Emergency Project from the Science and Technology Commission of Chongqing(cstc2020jscx-fyzx0053,cstc2020jscx-dxwtB0050 to A-L.H.)the Emergency Project for Novel Coronavirus Pneumonia from the Chongqing Medical University(CQMUNCP0302 to K.W.).
文摘Dear Editor,Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is a major public health issue.The spike(S)protein mutation D614G became dominant in SARS-CoV-2 during a global pandemic,which displayed increased infectivity.1 Entry of a virus into host cells is one of the most critical steps in the viral life cycle.Since blockade of the entry process is a promising therapeutic option for COVID-19,research attention has been focused on the discovery of viral entry inhibitors.Although SARS-CoV-2 entry inhibitor development is very attractive,no candidates have progressed into clinical trials yet.
文摘Apolipoprotein E(APOE)plays a pivotal role in lipid including cholesterol metabolism.The APOE 4(APOE4)allele is a major genetic risk factor for Alzheimer's and cardiovascular diseases.Although APOE has recently been associated with increased susceptibility to infections of several viruses,whether and how APOEand its isoforms affect SARS-CoV-2 infection remains unclear.Here,we show that serum concentrations of APOE correlate inversely with levels of cytokine/chemokine in 73 COVID-19 patients.Utilizing multiple protein interaction assays,we demonstrate that APOE3 and APOE4 interact with the SARS-CoV-2 receptor ACE2;and APOE/ACE2 interactions require zinc metallopeptidase domain of ACE2,a key docking site for SARS-CoV-2 Spike protein.In addition,immuno-imaging assays using confocal,super-resolution,and transmission electron microscopies reveal that both APOE3 and APOE4 reduce ACE2/Spikemediated viral entry into cells.Interestingly,while having a comparable binding affinity to ACE2,APOE4 inhibits viral entry to a lesser extent compared to APOE3,which is likely due to APOE4's more compact structure and smaller spatial obstacle to compete against Spike binding to ACE2.Furthermore,APOE e4 carriers clinically correlate with increased SARS-CoV-2 infection and elevated serum inflammatory factors in 142 COVID-19 patients assessed.Our study suggests a regulatory mechanism underlying SARS-CoV-2 infection through APOE interactions with ACE2,which may explain in part increased COVID-19 infection and disease severity in APOE e4 carriers.
基金This work was supported by National Natural Science Foundation of China(Grant No.81871656 and 8181101099 to J C)National Science and Technology Major Project(Grant No.2017ZX10202203 to AL H).
文摘In December 2019,the corona virus disease 2019(COVID-19)caused by novel coronavirus(SARS-CoV-2)emerged in Wuhan,China and rapidly spread worldwide.Few information on clinical features and immunological profile of COVID-19 in paediatrics.The clinical features and treatment outcomes of twelve paediatric patients confirmed as COVID-19 were analyzed.The immunological features of children patients was investigated and compared with twenty adult patients.The median age was 14.5-years(range from 0.64 to 17),and six of the patients were male.The average incubation period was 8 days.Clinically,cough(9/12,75%)and fever(7/12,58.3%)were the most common symptoms.Four patients(33.3%)had diarrhea during the disease.As to the immune profile,children had higher amount of total T cell,CD8t T cell and B cell but lower CRP levels than adults(P<0.05).Ground-glass opacity(GGO)and local patchy shadowing were the typical radiological findings on chest CT scan.All patients received antiviral and symptomatic treatment and the symptom relieved in 3e4 days after admitted to hospital.The paediatric patients showed mild symptom but with longer incubation period.Children infected with SARS-CoV-2 had different immune profile with higher T cell amount and low inflammatory factors level,which might ascribed to the mild clinical symptom.We advise that nucleic acid test or examination of serum IgM/IgG antibodies against SARS-CoV-2 should be taken for children with exposure history regardless of clinical symptom.
基金supported by the National Natural Science Foundation of China(No.U20A20392)the 111 Project(No.D20028)+6 种基金Open Research Fund Program of the Key Laboratory of Molecular Biology for Infectious Diseases,China(No.CQMU202102 and CQMU202105)The Science and Technology Research Program of Chongqing Municipal Education Commission,China(No.KJZD-M202000401)The Natural Science Foundation Project of Chongqing,China(No.cstc2019jscx-dxwtBX0019)The Emergency Project from the Science&Technology Commission of Chongqing,China(No.cstc2020jscx-fyzx0053 and cstc2020jscx-dxwtB0050)Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University,the Emergency Project for Novel Coronavirus Pneumonia from the Chongqing Medical University,China(No.CQMUNCP0302)China Postdoctoral Science Foundation,China(No.2021M693924)Chongqing Postdoctoral Science Special Foundation,China(No.2010010005216630).
文摘Coronavirus disease 2019(COVID-19)is caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The Spike protein that mediates coronavirus entry into host cells is a major target for COVID-19 vaccines and antibody therapeutics.However,multiple variants of SARS-CoV-2 have emerged,which may potentially compromise vaccine effectiveness.Using a pseudovirus-based assay,we evaluated SARS-CoV-2 cell entry mediated by the viral Spike B.1.617 and B.1.1.7 variants.We also compared the neutralization ability of monoclonal antibodies from convalescent sera and neutralizing antibodies(NAbs)elicited by CoronaVac(inactivated vaccine)and ZF2001(RBD-subunit vaccine)against B.1.617 and B.1.1.7 variants.Our results showed that,compared to D614G and B.1.1.7 variants,B.1.617 shows enhanced viral entry and membrane fusion,as well as more resistant to antibody neutralization.These findings have important implications for understanding viral infectivity and for immunization policy against SARS-CoV-2 variants.
文摘Correction to:Cellular&Molecular Immunology https:/doi.org/10.1038/s41423-020-00605-4,published online 08 January 2021 The licence information was missing from this article and should havebeen CC-BY.The original article has been corrected.
