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Detection of HBV DNA integration in plasma cell-free DNA of different HBV diseases utilizing DNA capture strategy

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Virologica Sinica 2024年第4期39卷 655-666页

作者： Zerui Yang Jingyan Zeng Yueyue Chen Mengchun Wang Hongchun Luo ai-long huang Haijun Deng Yuan Hu Key Laboratory of Molecular Biology on Infectious Diseases Ministry of EducationChongqing Medical UniversityChongqing400016China Department of Infectious Diseases The First Affiliated Hospital of Chongqing Medical UniversityChongqing400016China

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 *** this study,we developed an improved strategy for detecting HBV DNA integration in plasma cfDNA,based on DNA probe capture and next-generation *** 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 *** 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,*** 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 *** majority of integration breakpoints(61.86%)are located in the gene-coding *** non-homologous end-joining(NHEJ)and microhomology-mediated end-joining(MMEJ)interactions occurred during HBV integration across the three different stages of liver *** 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.

关键词： Hepatitis B virus(HBV) DNA integration Cell-free DNA(cfDNA) DNA capture

Gene therapy that inhibits NF-κB results in apoptosis of human hepatocarcinoma by recombinant adenovirus

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World Journal of Gastroenterology 2006年第33期12卷 5287-5292页

作者： Tie-Jun Li Li-Ping Jia Xiao-Ling Gao ai-long huang The 9th Hospital of Chongqing Beibei 400700 Chongqing China Institute for Viral Hepatitis Chongqing Medical University Chongqing 400016 China

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.

关键词： NF-κB IκBα Adenovirus

P19 of Tomato Bushy Stunt Virus Suppresses RNA Silencing Induced by Short Hairpin RNA in Mammal Cells

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中国病毒学 2007年第3期22卷 199-206页

作者： Wei-xian CHEN　 Juan CHEN　 Zhen-zhen ZHANG　 ai-long huang　 Institute for Viral Hepatitis Key Laboratory on Infectious Disease of Ministry of Education Chongqing University of Medical Science Chongqing 400016 China

To counteract the immune system in parasitic hosts,some viruses encode proteins to suppress the RNA interference(RNai)*** this report,we established two RNai systems to be easily observed with strong and obvious *** function of the P19 of tomato bushy stunt virus,which suppresses RNai in mammal cells,was then studied using these two *** hairpin RNAs targeting green fluorescence protein(pshRNA-GFP)and firefly luciferase(pshRNA-luc)were designed and inserted into a eukaryotic transcriptional vector pTZU6+1,*** shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without *** GFP expression level was assayed by fluorescence microscopy,Western blotting and *** luciferase expression level was analyzed by the dual-luciferase assay *** designed in this study down-regulated the target gene specifically and efficiently,with a decrease of expression of both genes of about 70%,*** P19 was introduced into the RNai systems,the expression of both GFP and the luciferase were mostly recovered compared with the control *** RNai systems of GFP and luciferase were constructed successfully,demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNai effect induced by shRNA in mammal cells.

关键词：哺乳动物 RNA干涉番茄丛矮病毒 P19 RNA抑制

Emerging SARS-CoV-2 variants reduce neutralization sensitivity to convalescent sera and monoclonal antibodies

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Cellular & Molecular Immunology 2021年第4期18卷 1061-1063页

作者： Jie Hu Pai Peng Kai Wang Liang Fang Fei-yang Luo ai-shun Jin Bei-zhong Liu Ni Tang ai-long huang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious Diseasesthe Second Affiliated HospitalChongqing Medical UniversityChongqingChina Yong-Chuan Hospital Chongqing Medical UniversityChongqingChina Department of Immunology College of Basic MedicineChongqing Medical UniversityChongqingChina

Coronaviruses are enveloped,positive-stranded RNA viruses that contain the largest known RNA genomes to *** severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continues to circulate in the human population,mul... 详细信息

关键词： acute monoclonal respiratory

Increased immune escape of the new SARS-CoV-2 variant of concern Omicron

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Cellular & Molecular Immunology 2022年第2期19卷 293-295页

作者： Jie Hu Pai Peng Xiaoxia Cao Kang Wu Juan Chen Kai Wang Ni Tang ai-long huang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious Diseasesthe Second Affiliated HospitalChongqing Medical UniversityChongqingChina.

On 24 November,a new detected variant B.1.1.529 of SARS-CoV-2 by South Africa was reported to *** only 2 days,this variant was designated as“variant of concern”(VOC)and named as *** the past few weeks,Omicron had reported from more than 80 *** has been reported as the dominant SARS-CoV-2 in *** to the rapid spread of Omicron.A new wave of infection driven by Omicron is in progress.

关键词： immune infection Omi

Hepatitis B virus and hepatocellular carcinoma at the miRNA level

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World Journal of Gastroenterology 2011年第28期17卷 3353-3358页

作者： Zhen-Zhen Zhang Xiang Liu De-Qiang Wang Mai-Kun Teng Li-Wen Niu ai-long huang Zhi Liang Key Laboratory of Molecular Biology onInfectious Diseases Ministry of Education Institute for ViralHepatitis the Second Affiliated Hospital and Children＇s Hospitalof Chongqing Medical University Chongqing 400016 China KeyLaboratoryof Molecular Biology on Infectious Diseases Ministry of Educa-tion Institute for Viral Hepatitis the Second Affiliated HospitalChongqing Medical University Chougqing 400016 China Hefei National Labo-ratory for Physical Sciences at Microscale and School of LifeScience University of Science and Technology of China HefeiAnhui Province 230027 China

aiM: To study Hepatitis B virus (HBV) infection and its association with hepatocellular carcinoma (HCC) at the miRNA ***: 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 *** miRNA levels were examined using microarray *** 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 *** compared the miRNA expression alterations during HBV infection with those in HCC patients,by analyzing miRNA expression change profiles ***: Seventy-seven and 48 miRNAs were differentially expressed during acute and chronic HBV infection,*** 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 *** showed inverse changes during the two phases of infection;downregulated in the acute infection and upregulated in the chronic *** results imply that common and specific mechanisms exist at the miRNA level during acute and chronic HBV ***,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 duri

关键词： Hepatitis B virus Hepatocellular carcinoma MiRNA

Identification of bis-benzylisoquinoline alkaloids as SARS-CoV-2 entry inhibitors from a library of natural products

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Signal Transduction and Targeted Therapy 2021年第4期6卷 1113-1115页

作者： Chang-long He Lu-Yi huang Kai Wang Chen-Jian Gu Jie Hu Gui-Ji Zhang Wei Xu You-Hua Xie Ni Tang ai-long huang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious DiseasesThe Second Affiliated HospitalChongqing Medical UniversityChongqingChina Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) Department of Medical Microbiology and ParasitologySchool of Basic Medical SciencesShanghai Medical CollegeFudan UniversityShanghaiChina Children's Hospital Fudan UniversityShanghaiChina

Dear Editor,Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is a major public health *** 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 *** blockade of the entry process is a promising therapeutic option for COVID-19,research attention has been focused on the discovery of viral entry *** SARS-CoV-2 entry inhibitor development is very attractive,no candidates have progressed into clinical trials yet.

关键词： acute respiratory attractive

Changes in the humoral immunity response in SARS-CoV-2 convalescent patients over 8 months

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Cellular & Molecular Immunology 2021年第2期18卷 490-491页

作者： Pai Peng Jie Hu Hai-jun Deng Bei-zhong Liu Liang Fang Kai Wang Ni Tang ai-long huang Key Laboratory of Molecular Biology for Infectious Diseases(Ministry of Education) Institute for Viral HepatitisDepartment of Infectious DiseasesThe Second Affiliated HospitalChongqing Medical UniversityChongqingChina Yong-Chuan Hospital Chongqing Medical UniversityChongqing402160China

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 *** is a current need to follow the maintenance of specific antibodies against SARS-CoV-2.

关键词： immunity acute respiratory

A novel phenotypic assay of hepatitis B virus polymerase with extensive site-specific mutagenesis

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Virologica Sinica 2017年第2期32卷 167-170页

作者： ya liu ying-ying luo xue-fei cai quan-xin long chun-yang gan liu-qing yang haitao guo ai-long huang wen-lu zhang jie-li hu institute for viral hepatitis key laboratory of molecular biology on infectious diseasesministry of educationthe second affiliated hospital of chongqing medical universityChongqing 400016China department of microbiology and immunology indiana university school of medicine indianapolis46202 USA collaborative innovation center for diagnosis and treatment of infectious diseases(ccid) Hangzhou 310058China

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... 详细信息

关键词： In HBVDNA FSR A novel phenotypic assay of hepatitis B virus polymerase with extensive site-specific mutagenesis LAM