AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine. METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support prod...AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine. METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization. RESULTS: IFN-γat 0.1 to 5μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells. At 5μg/L, IFN-γalso suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γshowed no additive effect, sequential treatment first with lamivudine and then IFN-γwas found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2μmol/L lamivudine for two days, followed by 1μg/L IFN-γfor another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2μmol/L lamivudine for two days, followed by 5μg/L IFN-γfor six days showed a 72% reduction in HBV cccDNA pool. CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γand lamivudine, especially in IFN-αnon-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.展开更多
AIM： To evaluate the anticancer property of the dried latex （DL） of Calotropis procera, a tropical medicinal plant, in the X15-myc transgenic mouse model of hepatocellular carcinoma and to elucidate its mechanism o...AIM： To evaluate the anticancer property of the dried latex （DL） of Calotropis procera, a tropical medicinal plant, in the X15-myc transgenic mouse model of hepatocellular carcinoma and to elucidate its mechanism of action in cell culture. METHODS： The young transgenic mice were orally fed with the aqueous suspension of DL （400 mg/kg for 5 d/wk） for 15 wk and their liver was examined for histopathological changes at 20 wk. Serum levels of vascu- lar endothelial growth factor （VEGF） were also measured in these animals. To characterize the active fraction, DL was extracted with petroleum ether followed by methanol. The methanolic extract was sub-fractionated on a silica gel G column using a combination of non-polar and polar solvents and eleven fractions were obtained. Each fraction was analysed for cytotoxic effect on hepatoma （Huh7） and non-hepatoma （COS-1） cell lines and nontransformed hepatocytes （AML12） using tetrazolium （MTT） assay. Finally, the mechanism of cell death was investigated by measuring the levels of Bcl2, caspase 3 and DNA fragmentation. RESULTS： DL treatment of mice showed a complete protection against hepatocarcinogenesis. No adverse effect was observed in these animals. The serum VEGF level was significantly lowered in the treated mice as compared to control animals. Cell culture studies revealed that the methanolic extract of DL as well as its fraction 8 induced extensive cell death in both Huh-7 and COS-1 cells while AML12 cells were spared. This was accompanied by extensive fragmentation of DNA in Huh-7 and COS-1 cells. No change in the levels of canonical markers of apoptosis such as Bcl2 and caspase 3 was observed. CONCLUSION： DL of C. procera has the potential for anti-cancer therapy due to its differentJable targets and non-interference with regular pathway of apoptosis.展开更多
The flower of Butea monosperma(Lam.)(Fabaceae)has been used in traditional Indian medicine in the treatment of many ailments including liver disorders.To understand the pharmacological basis of its beneficial effects,...The flower of Butea monosperma(Lam.)(Fabaceae)has been used in traditional Indian medicine in the treatment of many ailments including liver disorders.To understand the pharmacological basis of its beneficial effects,the extracts of dried flowers in water,methanol,butanol,ethyl acetate and acetone were evaluated for free radical scavenging and pro-apoptotic activities in cell cultures(human hepatoma Huh-7 cell line and immortalized AML-12 mouse hepatocytes).Butrin and butein-the active constituents of flower extracts-were used as reference molecules.The levels of cell injury markers like lactate dehydrogenase,glutathione and lipid peroxidation and primary antioxidant enzymes glutathione S-transferase and catalase were also measured.The aqueous and butanolic extracts exhibited better 2,2-diphenyl-1-picrylhydrazyl scavenging and cytotoxic activities in hepatoma cells than in immortalized hepatocytes.Interestingly,butein inhibited 2,2-diphenyl-1-picrylhydrazyl radical better than butrin.The aqueous and butanolic extracts were further investigated for hepatoprotection against carbon tertrachloride-induced biochemical changes and cell death.Both extracts,just as butrin and butein,significantly reversed the cellular glutathione levels and lipid peroxidation,and glutathione–S-transferase activity.Lactate dehydrogenase leakage and cell death were also prevented.However,only butein revived the catalase activity.Thus,the butein content of Butea monosperma flower extracts is important for free radical scavenging activity,apoptotic cell death and protection against oxidative injury in hepatic cells.展开更多
AIM: To study host gene expression and numberof immune cells in liver tissues from patients with fulminant hepatitis E(FH-E).METHODS: Microarray-based expression profiling was done using Illumina Human WG-6_v3_Bead Ch...AIM: To study host gene expression and numberof immune cells in liver tissues from patients with fulminant hepatitis E(FH-E).METHODS: Microarray-based expression profiling was done using Illumina Human WG-6_v3_Bead Chip arrays on post-mortem liver tissue from 5 patients with FH-E,and compared with similar tissue from 6 patients with fulminant hepatitis B(FH-B; disease controls) and normal liver tissue from 6 persons.Differential expression was defined as ≥ 2.0-fold change with Benjamini-Hochberg false discovery rate below 0.05 using t-test in liver tissue from FH-B and FH-E,than healthy liver tissue.For some genes that showed differential expression in FH-E,microarray data were validated using quantitative reverse transcription PCR.Differentially expressed gene lists were then subjected to "Gene Ontology" analysis for biological processes,and pathway analysis using Bio Carta database on the DAVID server.In addition,tissue sections were stained for CD4+,CD8+ and CD56+ cells using indirect immunohistochemistry; cells staining positive for each of these markers were counted and compared between groups.RESULTS: Compared to normal livers,those from patients with FH-E and FH-B showed differential expression of 3377 entities(up-regulated 1703,downregulated 1674) and 2572 entities(up 1164,down 1408),respectively.This included 2142(up 896,down 1246) entities that were common between the two sets; most of these belonged to metabolic,hemostatic and complement pathways,which are active in normal livers.Gene expression data from livers of patients with FH-E but not those of FH-B showed activation of several immune response pathways,particularly those involving cytotoxic T cells.The fold-change values of m RNA for selected genes in livers from FH-E than in normal liver tissue determined using quantitative reverse transcription PCR showed excellent concordance with microarray analysis.At immunohistochemistry,CD8+ T cells showed an increase in liver biopsies from both FH-E [median 53.4 per arbitrary unit area(range 31.2-99.9)] and FH-B [median 49.3(19.3-51.0); P = 0.005] compared to control liver tissue [median 6.9(3.1-14.9)].CONCLUSION: FH-E patients show CD8+ T cell infiltration and increased gene expression of cytotoxic T cell pathways in liver,suggesting a possible pathogenetic role for these cells.展开更多
Aim: To study the effect of androgen and antiandrogen on the level of androgen receptor (AR) mRNA. Methods: The total RNA was extracted from the prostate and analyzed by slot blot analysis. The blots were hybridized w...Aim: To study the effect of androgen and antiandrogen on the level of androgen receptor (AR) mRNA. Methods: The total RNA was extracted from the prostate and analyzed by slot blot analysis. The blots were hybridized with AR cDNA probe and 1A probe (internal control) and autoradiography was performed. The intensity of signal was measured with a densitometer and the ratio of AR RNA and 1A RNA was calculated. Results: Androgenic deprivation produced by castration decreased the weight of the prostate and increased the levels of AR mRNA. Treatment of the castrated rats with testostrone increased the weight of prostate and decreased the levels of AR mRNA. Treatment of normal rats with flutamide decreased the weight of the gland and increased the levels Of AR mRNA. Conclusion: Androgens produce proliferative effect on the prostate and negatively regulate the AR transcription.展开更多
Human endogenous retroviruses(HERVs) are the remains of ancient retroviruses that invaded our ancestors’ germline cell and were integrated into the genome. The expression of HERVs has always been a cause for concern ...Human endogenous retroviruses(HERVs) are the remains of ancient retroviruses that invaded our ancestors’ germline cell and were integrated into the genome. The expression of HERVs has always been a cause for concern because of its association with various cancers and diseases. However, few previous studies have focused on specific activation of HERVs by viral infections. Our previous study has shown that dengue virus type 2(DENV-2) infection induces the transcription of a large number of abnormal HERVs loci;therefore, the purpose of this study was to explore the relationship between exogenous viral infection and HERV activation further. In this study, we retrieved and reanalyzed published data on 21 transcriptomes of human cells infected with various viruses. We found that infection with different viruses could induce transcriptional activation of HERV loci. Through the comparative analysis of all viral datasets, we identified 43 key HERV loci that were up-regulated by DENV-2, influenza A virus, influenza B virus, Zika virus, measles virus, and West Nile virus infections. Furthermore, the neighboring genes of these HERVs were simultaneously up-regulated, and almost all such neighboring genes were interferon-stimulated genes(ISGs), which are enriched in the host’s antiviral immune response pathways. Our data supported the hypothesis that activation of HERVs, probably via an interferon-mediated mechanism, plays an important role in innate immunity against viral infections.展开更多
Tumor angiogenesis is the uncontrolled growth of blood vessels in tumors,serving to supply nutrients and oxygen,and remove metabolic wastes. Kaposi’s sarcoma (KS),a multifocal angioproliferative disorder characterize...Tumor angiogenesis is the uncontrolled growth of blood vessels in tumors,serving to supply nutrients and oxygen,and remove metabolic wastes. Kaposi’s sarcoma (KS),a multifocal angioproliferative disorder characterized by spindle cell proliferation,neo-angiogenesis,inflammation,and edema,is associated with infection by Kaposi's sarcoma-associated herpesvirus (KSHV). Recent studies indicate that KSHV infection directly promotes angiogenesis and inflammation through an autocrine and paracrine mechanism by inducing pro-angiogenic and pro-inflammatory cytokines. Many of these cytokines are also expressed in KS lesions,implicating a direct role of KSHV in the pathogenesis of this malignancy. Several KSHV genes are involved in KSHV-induced angiogenesis. These studies have provided insights into the pathogenesis of KS,and identified potential therapeutic targets for this malignancy.展开更多
Dear Editor,It has been more than 1 year since China reported the first case of African swine fever (ASF) infection in August2018, and the epidemic situation remains severe (China News Service 2019). According to repo...Dear Editor,It has been more than 1 year since China reported the first case of African swine fever (ASF) infection in August2018, and the epidemic situation remains severe (China News Service 2019). According to reports from the Ministry of Agriculture and Rural Affairs, China has reported 160 cases of ASF, which resulted in nearly 1.2 million pigs being killed, as of November 21, 2019 (China News Service 2019).展开更多
Genome sequencing has shown strong capabilities in the initial stages of the COVID-19 pandemic such as pathogen identification and virus preliminary tracing.While the rapid acquisition of SARS-Co V-2 genome from clini...Genome sequencing has shown strong capabilities in the initial stages of the COVID-19 pandemic such as pathogen identification and virus preliminary tracing.While the rapid acquisition of SARS-Co V-2 genome from clinical specimens is limited by their low nucleic acid load and the complexity of the nucleic acid background.To address this issue,we modified and evaluated an approach by utilizing SARS-Co V-2-specific amplicon amplification and Oxford Nanopore Prometh ION platform.This workflow started with the throat swab of the COVID-19 patient,combined reverse transcript PCR,and multi-amplification in one-step to shorten the experiment time,then can quickly and steadily obtain high-quality SARS-Co V-2 genome within 24 h.A comprehensive evaluation of the method was conducted in 42 samples:the sequencing quality of the method was correlated well with the viral load of the samples;high-quality SARS-Co V-2 genome could be obtained stably in the samples with Ct value up to 39.14;data yielding for different Ct values were assessed and the recommended sequencing time was 8 h for samples with Ct value of less than 20;variation analysis indicated that the method can detect the existing and emerging genomic mutations as well;Illumina sequencing verified that ultra-deep sequencing can greatly improve the single read error rate of Nanopore sequencing,making it as low as 0.4/10,000 bp.In summary,high-quality SARS-Co V-2 genome can be acquired by utilizing the amplicon amplification and it is an effective method in accelerating the acquisition of genetic resources and tracking the genome diversity of SARSCo V-2.展开更多
MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress th...MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress the innate and adaptive host immunity,modulate cellular signaling pathways,and regulate the expression of cellular and viral genes. These functions facilitate viral acute and persistent infections,and have profound effects on the host cell survival and disease progression. Here,we discuss the miRNAs encoded by herpesviruses,and their regulatory roles involved in virus-host interactions.展开更多
Human respiratory syncytial virus(RSV)is a severe threat to children and a main cause of acute lower respiratory tract infections.Nevertheless,the intra-host evolution and inter-regional diffusion of RSV are little kn...Human respiratory syncytial virus(RSV)is a severe threat to children and a main cause of acute lower respiratory tract infections.Nevertheless,the intra-host evolution and inter-regional diffusion of RSV are little known.In this study,we performed a systematic surveillance in hospitalized children in Hubei during 2020–2021,in which 106 RSV-positive samples were detected both clinically and by metagenomic next generation sequencing(mNGS).RSV-A and RSV-B groups co-circulated during surveillance with RSV-B being predominant.About 46 high-quality genomes were used for further analyses.A total of 163 intra-host nucleotide variation(iSNV)sites distributed in 34 samples were detected,and glycoprotein(G)gene was the most enriched gene for iSNVs,with non-synonymous substitutions more than synonymous substitutions.Evolutionary dynamic analysis showed that the evolutionary rates of G and NS2 genes were higher,and the population size of RSV groups changed over time.We also found evidences of inter-regional diffusion from Europe and Oceania to Hubei for RSV-A and RSV-B,respectively.This study highlighted the intra-host and inter-host evolution of RSV,and provided some evi-dences for understanding the evolution of RSV.展开更多
Dear Editor,African swine fever(ASF)is a severe haemorrhagic fever in domestic pigs and wild boar with extremely high mortality rate.It is cataloged as a notifiable disease by the World Organization for Animal Health(...Dear Editor,African swine fever(ASF)is a severe haemorrhagic fever in domestic pigs and wild boar with extremely high mortality rate.It is cataloged as a notifiable disease by the World Organization for Animal Health(OIE).The etiological agent that causes the highly lethal disease is the African swine fever virus(ASFV)(Sanchez-Vizcaino) et al.展开更多
Correction to:Virologica Sinica(2021)36:901-912 https://doi.Org/10.1007/sl 2250-021-00378-8 The original version of article contains errors in Figure 4,Supplementary Figure S2,Supplementary Table S2,and the correspond...Correction to:Virologica Sinica(2021)36:901-912 https://doi.Org/10.1007/sl 2250-021-00378-8 The original version of article contains errors in Figure 4,Supplementary Figure S2,Supplementary Table S2,and the corresponding description in part 3 of“Results”section.展开更多
AIM: To investigate if correction of hypovitaminosis D before initiation of Peg-interferon-alpha/ribavirin(Peg IFN/RBV) therapy could improve the efficacy of Peg IFN/RBV in previously null-responder patients with chro...AIM: To investigate if correction of hypovitaminosis D before initiation of Peg-interferon-alpha/ribavirin(Peg IFN/RBV) therapy could improve the efficacy of Peg IFN/RBV in previously null-responder patients with chronic genotype 1 or 4 hepatitis C virus(HCV) infection.METHODS:Genotype 1 or 4 HCV-infected patients with null response to previous Peg IFN/RBV treatment and with hypovitaminosis D(<30 ng/m L)prospectively received cholecalciferol 100000 IU per week for 4 wk[from week-4(W-4)to W0],followed by 100000 IUper month in combination with Peg IFN/RBV for 12 mo(from W0 to W48).The primary outcome was the rate of early virological response defined by an HCV RNA<12 IU/m L after 12 wk Peg IFN/RBV treatment.RESULTS:A total of 32 patients were included,19(59%)and 13(41%)patients were HCV genotype1 and 4,respectively.The median baseline vitamin D level was 15 ng/m L(range:7-28).In modified intention-to-treat analysis,29 patients who received at least one dose of Peg IFN/RBV were included in the analysis.All patients except one normalized their vitamin D serum levels.The rate of early virologic response was 0/29(0%).The rate of HCV RNA<12IU/m L after 24 wk of Peg IFN/RBV was 1/27(4%).The safety profile was favorable.CONCLUSION:Addition of vitamin D to Peg IFN/RBV does not improve the rate of early virologic response in previously null-responders with chronic genotype 1or 4 HCV infection.展开更多
Objective:Acupuncture is effective in the treatment of polycystic ovary syndrome with insulin resistance(PCOS-IR),but the underlying mechanism is still unclear.In this study,we further investigated the regulatory effe...Objective:Acupuncture is effective in the treatment of polycystic ovary syndrome with insulin resistance(PCOS-IR),but the underlying mechanism is still unclear.In this study,we further investigated the regulatory effects of electroacupuncture(EA)on IR,histone acetylation,and oxidative stress in PCOS-IR rats.Materials and Methods:PCOS-IR rats were induced with letrozole combined with a high-fat diet.EA at Yinsanzhen(CV4,SP6,and ST29)was performed daily for 3 weeks.Fasting blood glucose and area under the curve were assessed through a glucose tolerance test.Ovarian morphology was observed through hematoxylin and eosin staining.The levels of glucose and lipid metabolism and reproductive endocrine parameters in the rats were evaluated with a test kit.RNA-seq and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the gene expression.Results:E A significantly improved the regularity of the estrous cycle,ovarian mo rphology,glucose-lipid metab olism,and repro ductive endocrine system in PCOS-IR rats.RNA-seq revealed that EA had certain regulato ry effects on the HD AC 1,NRF1,Mfnl,OPA1,Drpl(DnmlI),and Fis1 genes.RT-qPCR further verified that EA significantly upregulated the mRNA expression of HDAC1,histone H4,OPA1,and Fis1 and tended to decrease the level of the AKT mRNA.Conclusions:EA can improve the pathological manifestations of the ovary,glucose-lipid metabolism,and reproductive endocrine system in PCOS-IR model rats.EA can increase the expression of HDAC1 and histone H4.In addition,EA can increase the expression of OPA1 and Fis1,decrease the expression of AKT,to reduce oxidative stress and improve IR.展开更多
HIV-1 co-receptor tropism is central for understanding the transmission and pathogenesis of HIV-1 infection. We performed a genome-wide comparison between the adaptive evolution of R5 and X4 variants from HIV-1 subtyp...HIV-1 co-receptor tropism is central for understanding the transmission and pathogenesis of HIV-1 infection. We performed a genome-wide comparison between the adaptive evolution of R5 and X4 variants from HIV-1 subtypes B and C. The results showed that R5 and X4 variants experienced differential evolutionary patterns and different HIV-1 genes encountered various positive selection pressures, suggesting that complex selection pressures are driving HIV-1 evolution. Compared with other hypervariable regions of Gp120, significantly more positively selected sites were detected in the V3 region of subtype B X4 variants, V2 region of subtype B R5 variants, and V1 and V4 regions of subtype C X4 variants, indicating an association of positive selection with co-receptor recognition/binding. Intriguingly, a significantly higher proportion (33.3% and 55.6%, P【0.05) of positively selected sites were identified in the C3 region than other conserved regions of Gp120 in all the analyzed HIV-1 variants, indicating that the C3 region might be more important to HIV-1 adaptation than previously thought. Approximately half of the positively selected sites identified in the env gene were identical between R5 and X4 variants. There were three common positively selected sites (96, 113 and 281) identified in Gp41 of all X4 and R5 variants from subtypes B and C. These sites might not only suggest a functional importance in viral survival and adaptation, but also imply a potential cross-immunogenicity between HIV-1 R5 and X4 variants, which has important implications for AIDS vaccine development.展开更多
Despite the long journey undertaken by mankind to confront the diseases caused by viruses,people were unaware of the existence of viruses until the closing years of the 19th century,when the Russian biologist Dmitri I...Despite the long journey undertaken by mankind to confront the diseases caused by viruses,people were unaware of the existence of viruses until the closing years of the 19th century,when the Russian biologist Dmitri Ivanovsky demonstrated that tobacco plants could be infected by a toxin smaller than bacteria,followed by which the Dutch microbiologist Martinus Beijerinck termed the toxin as a contagium vivum fluidum （soluble living germ） and re-introduced the word ＂virus＂ （derived from the Latin word ＂virus＂ meaning poison; the modern meaning is an ＂agent that causes infectious disease＂ that was first recorded in 1728）.展开更多
Dear Editors,Here, we report a novel HIV-1 intra-circulating recombinant form 01_AE （intra-CRF01_AE： CRF01-1AE/CRF01-6AE） composed of CRF01_AE transmission clusters 1 and 6, which was identified in a heterosexual m...Dear Editors,Here, we report a novel HIV-1 intra-circulating recombinant form 01_AE （intra-CRF01_AE： CRF01-1AE/CRF01-6AE） composed of CRF01_AE transmission clusters 1 and 6, which was identified in a heterosexual male from Fujian, with one breakpoint observed in the vif gene. The CRF01_AE region I （HXB2： 868-5184） of the recombinant clustered with CRF01_AE transmission cluster 1, which is mainly circulating among injection drug users （IDUs） and heterosexuals in southern and southwestern China, with the support of a 100% bootstrap value. The CRF01_AE region II （HXB2： 5185-9605） of the recombinant clustered with CRF01_AE transmission cluster 6, which mainly prevails among heterosexuals in southern China, with the support of a 100% bootstrap value. To our knowledge, this is the first detection of a novel HIV-1 intra-CRF01_AE recombinant form （CRF01-1_AE/CRF01-6_ AE） in Fujian, China, which indicates ongoing active HIV-1 transmission among heter- osexuals in this region.展开更多
Chronic human immunodeficiency virus(HIV) infection not only causes a gradual loss of CD4~+T cells but also leads to a disturbance of the T cell receptor(TCR) repertoire. In people living with HIV(PLWH), monitoring TC...Chronic human immunodeficiency virus(HIV) infection not only causes a gradual loss of CD4~+T cells but also leads to a disturbance of the T cell receptor(TCR) repertoire. In people living with HIV(PLWH), monitoring TCR repertoire is challenged by the inconsistency of complementarity determining region 3(CDR3) and limited cell numbers in clinical samples. Thus, a quantitative method is necessary for monitoring the TCR repertoire in PLWH.We characterized the TCR V-J pairing profile of na?ve and memory CD4~+T cells in healthy donors, HIV-infected antiretroviral therapy(ART)-na?ve patients and long-term(over5 years) ART-experienced patients by performing TCR sequencing. We developed a V-J index with 18 parameters which were subdivided into five categories(expression coverage, cumulative percentage of the top tenth percentile, diversity, intra-individual similarity and inter-individual similarity). In ART-na?ve patients, 14 of the 18 parameters were significantly altered.Long-term ART recovered ten parameters. The four unrecovered parameters were related to inter-individual similarity.Therefore, these findings indicate that long-term ART could only partially recover TCR V-J pairs and introduce newly impacted V-J pairs. Moreover, these results provide new insights into the V-J pairing of the TCR and into the disturbance of TCR repertoire in HIV infection.展开更多
基金Supported by a grant from the Dabur Research Foundation, India and a Senior Research Fellowship of the CSIR, Gov. of India (to MKP)
文摘AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine. METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization. RESULTS: IFN-γat 0.1 to 5μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells. At 5μg/L, IFN-γalso suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γshowed no additive effect, sequential treatment first with lamivudine and then IFN-γwas found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2μmol/L lamivudine for two days, followed by 1μg/L IFN-γfor another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2μmol/L lamivudine for two days, followed by 5μg/L IFN-γfor six days showed a 72% reduction in HBV cccDNA pool. CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γand lamivudine, especially in IFN-αnon-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.
基金Supported by the core grant of International Centre for Genetic Engineering and Biotechnology. New Delhi
文摘AIM： To evaluate the anticancer property of the dried latex （DL） of Calotropis procera, a tropical medicinal plant, in the X15-myc transgenic mouse model of hepatocellular carcinoma and to elucidate its mechanism of action in cell culture. METHODS： The young transgenic mice were orally fed with the aqueous suspension of DL （400 mg/kg for 5 d/wk） for 15 wk and their liver was examined for histopathological changes at 20 wk. Serum levels of vascu- lar endothelial growth factor （VEGF） were also measured in these animals. To characterize the active fraction, DL was extracted with petroleum ether followed by methanol. The methanolic extract was sub-fractionated on a silica gel G column using a combination of non-polar and polar solvents and eleven fractions were obtained. Each fraction was analysed for cytotoxic effect on hepatoma （Huh7） and non-hepatoma （COS-1） cell lines and nontransformed hepatocytes （AML12） using tetrazolium （MTT） assay. Finally, the mechanism of cell death was investigated by measuring the levels of Bcl2, caspase 3 and DNA fragmentation. RESULTS： DL treatment of mice showed a complete protection against hepatocarcinogenesis. No adverse effect was observed in these animals. The serum VEGF level was significantly lowered in the treated mice as compared to control animals. Cell culture studies revealed that the methanolic extract of DL as well as its fraction 8 induced extensive cell death in both Huh-7 and COS-1 cells while AML12 cells were spared. This was accompanied by extensive fragmentation of DNA in Huh-7 and COS-1 cells. No change in the levels of canonical markers of apoptosis such as Bcl2 and caspase 3 was observed. CONCLUSION： DL of C. procera has the potential for anti-cancer therapy due to its differentJable targets and non-interference with regular pathway of apoptosis.
基金supported by grant no.90(0039)/04/EMR-II of Council of Scientific and Industrial Research,Government of India,New Delhi.Anuradha Sehrawat has been a Research Associate of Department of Biotechnology,Government of India,during the period of this study。
文摘The flower of Butea monosperma(Lam.)(Fabaceae)has been used in traditional Indian medicine in the treatment of many ailments including liver disorders.To understand the pharmacological basis of its beneficial effects,the extracts of dried flowers in water,methanol,butanol,ethyl acetate and acetone were evaluated for free radical scavenging and pro-apoptotic activities in cell cultures(human hepatoma Huh-7 cell line and immortalized AML-12 mouse hepatocytes).Butrin and butein-the active constituents of flower extracts-were used as reference molecules.The levels of cell injury markers like lactate dehydrogenase,glutathione and lipid peroxidation and primary antioxidant enzymes glutathione S-transferase and catalase were also measured.The aqueous and butanolic extracts exhibited better 2,2-diphenyl-1-picrylhydrazyl scavenging and cytotoxic activities in hepatoma cells than in immortalized hepatocytes.Interestingly,butein inhibited 2,2-diphenyl-1-picrylhydrazyl radical better than butrin.The aqueous and butanolic extracts were further investigated for hepatoprotection against carbon tertrachloride-induced biochemical changes and cell death.Both extracts,just as butrin and butein,significantly reversed the cellular glutathione levels and lipid peroxidation,and glutathione–S-transferase activity.Lactate dehydrogenase leakage and cell death were also prevented.However,only butein revived the catalase activity.Thus,the butein content of Butea monosperma flower extracts is important for free radical scavenging activity,apoptotic cell death and protection against oxidative injury in hepatic cells.
基金Supported by National Institutes of Health,Bethesda,United States,No.5R01AI076192
文摘AIM: To study host gene expression and numberof immune cells in liver tissues from patients with fulminant hepatitis E(FH-E).METHODS: Microarray-based expression profiling was done using Illumina Human WG-6_v3_Bead Chip arrays on post-mortem liver tissue from 5 patients with FH-E,and compared with similar tissue from 6 patients with fulminant hepatitis B(FH-B; disease controls) and normal liver tissue from 6 persons.Differential expression was defined as ≥ 2.0-fold change with Benjamini-Hochberg false discovery rate below 0.05 using t-test in liver tissue from FH-B and FH-E,than healthy liver tissue.For some genes that showed differential expression in FH-E,microarray data were validated using quantitative reverse transcription PCR.Differentially expressed gene lists were then subjected to "Gene Ontology" analysis for biological processes,and pathway analysis using Bio Carta database on the DAVID server.In addition,tissue sections were stained for CD4+,CD8+ and CD56+ cells using indirect immunohistochemistry; cells staining positive for each of these markers were counted and compared between groups.RESULTS: Compared to normal livers,those from patients with FH-E and FH-B showed differential expression of 3377 entities(up-regulated 1703,downregulated 1674) and 2572 entities(up 1164,down 1408),respectively.This included 2142(up 896,down 1246) entities that were common between the two sets; most of these belonged to metabolic,hemostatic and complement pathways,which are active in normal livers.Gene expression data from livers of patients with FH-E but not those of FH-B showed activation of several immune response pathways,particularly those involving cytotoxic T cells.The fold-change values of m RNA for selected genes in livers from FH-E than in normal liver tissue determined using quantitative reverse transcription PCR showed excellent concordance with microarray analysis.At immunohistochemistry,CD8+ T cells showed an increase in liver biopsies from both FH-E [median 53.4 per arbitrary unit area(range 31.2-99.9)] and FH-B [median 49.3(19.3-51.0); P = 0.005] compared to control liver tissue [median 6.9(3.1-14.9)].CONCLUSION: FH-E patients show CD8+ T cell infiltration and increased gene expression of cytotoxic T cell pathways in liver,suggesting a possible pathogenetic role for these cells.
文摘Aim: To study the effect of androgen and antiandrogen on the level of androgen receptor (AR) mRNA. Methods: The total RNA was extracted from the prostate and analyzed by slot blot analysis. The blots were hybridized with AR cDNA probe and 1A probe (internal control) and autoradiography was performed. The intensity of signal was measured with a densitometer and the ratio of AR RNA and 1A RNA was calculated. Results: Androgenic deprivation produced by castration decreased the weight of the prostate and increased the levels of AR mRNA. Treatment of the castrated rats with testostrone increased the weight of prostate and decreased the levels of AR mRNA. Treatment of normal rats with flutamide decreased the weight of the gland and increased the levels Of AR mRNA. Conclusion: Androgens produce proliferative effect on the prostate and negatively regulate the AR transcription.
基金This work was supported by the China Ministry of Science and Technology Key Research&Development Program(No.2016YFC1200800)the National Natural Science Foundation of China(No.91631110)the National Mega Project on Major Infectious Disease Prevention(No.2017ZX10103005-005)。
文摘Human endogenous retroviruses(HERVs) are the remains of ancient retroviruses that invaded our ancestors’ germline cell and were integrated into the genome. The expression of HERVs has always been a cause for concern because of its association with various cancers and diseases. However, few previous studies have focused on specific activation of HERVs by viral infections. Our previous study has shown that dengue virus type 2(DENV-2) infection induces the transcription of a large number of abnormal HERVs loci;therefore, the purpose of this study was to explore the relationship between exogenous viral infection and HERV activation further. In this study, we retrieved and reanalyzed published data on 21 transcriptomes of human cells infected with various viruses. We found that infection with different viruses could induce transcriptional activation of HERV loci. Through the comparative analysis of all viral datasets, we identified 43 key HERV loci that were up-regulated by DENV-2, influenza A virus, influenza B virus, Zika virus, measles virus, and West Nile virus infections. Furthermore, the neighboring genes of these HERVs were simultaneously up-regulated, and almost all such neighboring genes were interferon-stimulated genes(ISGs), which are enriched in the host’s antiviral immune response pathways. Our data supported the hypothesis that activation of HERVs, probably via an interferon-mediated mechanism, plays an important role in innate immunity against viral infections.
基金The Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences (0702121YJ1)Open Research Fund Program of the State Key Laboratory of Virology of China (2007013)+1 种基金A Type B Outstanding Abroad Young Scientist Award (30328001) from the National Science Foundation of Chinagrants from the National Institutes of Health (CA096512,CA124332,CA119889 and DE017333)
文摘Tumor angiogenesis is the uncontrolled growth of blood vessels in tumors,serving to supply nutrients and oxygen,and remove metabolic wastes. Kaposi’s sarcoma (KS),a multifocal angioproliferative disorder characterized by spindle cell proliferation,neo-angiogenesis,inflammation,and edema,is associated with infection by Kaposi's sarcoma-associated herpesvirus (KSHV). Recent studies indicate that KSHV infection directly promotes angiogenesis and inflammation through an autocrine and paracrine mechanism by inducing pro-angiogenic and pro-inflammatory cytokines. Many of these cytokines are also expressed in KS lesions,implicating a direct role of KSHV in the pathogenesis of this malignancy. Several KSHV genes are involved in KSHV-induced angiogenesis. These studies have provided insights into the pathogenesis of KS,and identified potential therapeutic targets for this malignancy.
基金supported by the Research Project of African Swine Fever of Chinese Academy of Sciences (KJZD-SWL06)the National Natural Science Foundation of China (31941015)+3 种基金the National Key R&D Program of China (2016YFC1200800&2018YFC0840402)the China Mega-Project for Infectious Disease(2017ZX10103005-005)the State Key Laboratory of Veterinary Biotechnology Research Fund (SKLVBF201902)supported by Youth Innovation Promotion Association of CAS (2019091)。
文摘Dear Editor,It has been more than 1 year since China reported the first case of African swine fever (ASF) infection in August2018, and the epidemic situation remains severe (China News Service 2019). According to reports from the Ministry of Agriculture and Rural Affairs, China has reported 160 cases of ASF, which resulted in nearly 1.2 million pigs being killed, as of November 21, 2019 (China News Service 2019).
基金supported by grants from the Foundation for National Mega Project on Major Infectious Disease Prevention(grant number 2017ZX10103005-005)National Key Research and Development Program of China(2020YFC0845800 and 2020YFC0845600)the National Natural Science Foundation of China(31970548 and 91631110)。
文摘Genome sequencing has shown strong capabilities in the initial stages of the COVID-19 pandemic such as pathogen identification and virus preliminary tracing.While the rapid acquisition of SARS-Co V-2 genome from clinical specimens is limited by their low nucleic acid load and the complexity of the nucleic acid background.To address this issue,we modified and evaluated an approach by utilizing SARS-Co V-2-specific amplicon amplification and Oxford Nanopore Prometh ION platform.This workflow started with the throat swab of the COVID-19 patient,combined reverse transcript PCR,and multi-amplification in one-step to shorten the experiment time,then can quickly and steadily obtain high-quality SARS-Co V-2 genome within 24 h.A comprehensive evaluation of the method was conducted in 42 samples:the sequencing quality of the method was correlated well with the viral load of the samples;high-quality SARS-Co V-2 genome could be obtained stably in the samples with Ct value up to 39.14;data yielding for different Ct values were assessed and the recommended sequencing time was 8 h for samples with Ct value of less than 20;variation analysis indicated that the method can detect the existing and emerging genomic mutations as well;Illumina sequencing verified that ultra-deep sequencing can greatly improve the single read error rate of Nanopore sequencing,making it as low as 0.4/10,000 bp.In summary,high-quality SARS-Co V-2 genome can be acquired by utilizing the amplicon amplification and it is an effective method in accelerating the acquisition of genetic resources and tracking the genome diversity of SARSCo V-2.
基金The Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences (0702121YJ1)Open Research Fund Program of the State Key Laboratory of Virology of China(2007013)+1 种基金National Science Foundation of China (A Type B Outstanding Abroad Young Scientist Award)National Institutes of Health (CA096512,CA124332,CA119889 and DE017333)
文摘MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress the innate and adaptive host immunity,modulate cellular signaling pathways,and regulate the expression of cellular and viral genes. These functions facilitate viral acute and persistent infections,and have profound effects on the host cell survival and disease progression. Here,we discuss the miRNAs encoded by herpesviruses,and their regulatory roles involved in virus-host interactions.
基金National Key Research and Development Program of China(2018YFC1603803)National Natural Science Foun-dation of China(31970548)+2 种基金Knowledge Innovation Program of Wuhan-Basi Research(2022020801010519)Health Commission of Hubei Province(WJ 2021M262)Natural Science Fund of Hubei Province(2021CFA012).
文摘Human respiratory syncytial virus(RSV)is a severe threat to children and a main cause of acute lower respiratory tract infections.Nevertheless,the intra-host evolution and inter-regional diffusion of RSV are little known.In this study,we performed a systematic surveillance in hospitalized children in Hubei during 2020–2021,in which 106 RSV-positive samples were detected both clinically and by metagenomic next generation sequencing(mNGS).RSV-A and RSV-B groups co-circulated during surveillance with RSV-B being predominant.About 46 high-quality genomes were used for further analyses.A total of 163 intra-host nucleotide variation(iSNV)sites distributed in 34 samples were detected,and glycoprotein(G)gene was the most enriched gene for iSNVs,with non-synonymous substitutions more than synonymous substitutions.Evolutionary dynamic analysis showed that the evolutionary rates of G and NS2 genes were higher,and the population size of RSV groups changed over time.We also found evidences of inter-regional diffusion from Europe and Oceania to Hubei for RSV-A and RSV-B,respectively.This study highlighted the intra-host and inter-host evolution of RSV,and provided some evi-dences for understanding the evolution of RSV.
基金supported by grants from the National Key Research and Development Program of China(2018YFC1603803,2018YFC0840402 and 2016YFC1200800)the Research Project of African Swine Fever of Chinese Academy of Sciences(KJZD-SW-L06)
文摘Dear Editor,African swine fever(ASF)is a severe haemorrhagic fever in domestic pigs and wild boar with extremely high mortality rate.It is cataloged as a notifiable disease by the World Organization for Animal Health(OIE).The etiological agent that causes the highly lethal disease is the African swine fever virus(ASFV)(Sanchez-Vizcaino) et al.
文摘Correction to:Virologica Sinica(2021)36:901-912 https://doi.Org/10.1007/sl 2250-021-00378-8 The original version of article contains errors in Figure 4,Supplementary Figure S2,Supplementary Table S2,and the corresponding description in part 3 of“Results”section.
文摘AIM: To investigate if correction of hypovitaminosis D before initiation of Peg-interferon-alpha/ribavirin(Peg IFN/RBV) therapy could improve the efficacy of Peg IFN/RBV in previously null-responder patients with chronic genotype 1 or 4 hepatitis C virus(HCV) infection.METHODS:Genotype 1 or 4 HCV-infected patients with null response to previous Peg IFN/RBV treatment and with hypovitaminosis D(<30 ng/m L)prospectively received cholecalciferol 100000 IU per week for 4 wk[from week-4(W-4)to W0],followed by 100000 IUper month in combination with Peg IFN/RBV for 12 mo(from W0 to W48).The primary outcome was the rate of early virological response defined by an HCV RNA<12 IU/m L after 12 wk Peg IFN/RBV treatment.RESULTS:A total of 32 patients were included,19(59%)and 13(41%)patients were HCV genotype1 and 4,respectively.The median baseline vitamin D level was 15 ng/m L(range:7-28).In modified intention-to-treat analysis,29 patients who received at least one dose of Peg IFN/RBV were included in the analysis.All patients except one normalized their vitamin D serum levels.The rate of early virologic response was 0/29(0%).The rate of HCV RNA<12IU/m L after 24 wk of Peg IFN/RBV was 1/27(4%).The safety profile was favorable.CONCLUSION:Addition of vitamin D to Peg IFN/RBV does not improve the rate of early virologic response in previously null-responders with chronic genotype 1or 4 HCV infection.
基金supported by the National Natural Science Foundation of China(No.82074542)Joint Funds for the Innovation of Science and Technology,Fujian Province(No.2020Y9138)Fujian provincial health technology project(No.2023ZQNZD010)。
文摘Objective:Acupuncture is effective in the treatment of polycystic ovary syndrome with insulin resistance(PCOS-IR),but the underlying mechanism is still unclear.In this study,we further investigated the regulatory effects of electroacupuncture(EA)on IR,histone acetylation,and oxidative stress in PCOS-IR rats.Materials and Methods:PCOS-IR rats were induced with letrozole combined with a high-fat diet.EA at Yinsanzhen(CV4,SP6,and ST29)was performed daily for 3 weeks.Fasting blood glucose and area under the curve were assessed through a glucose tolerance test.Ovarian morphology was observed through hematoxylin and eosin staining.The levels of glucose and lipid metabolism and reproductive endocrine parameters in the rats were evaluated with a test kit.RNA-seq and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the gene expression.Results:E A significantly improved the regularity of the estrous cycle,ovarian mo rphology,glucose-lipid metab olism,and repro ductive endocrine system in PCOS-IR rats.RNA-seq revealed that EA had certain regulato ry effects on the HD AC 1,NRF1,Mfnl,OPA1,Drpl(DnmlI),and Fis1 genes.RT-qPCR further verified that EA significantly upregulated the mRNA expression of HDAC1,histone H4,OPA1,and Fis1 and tended to decrease the level of the AKT mRNA.Conclusions:EA can improve the pathological manifestations of the ovary,glucose-lipid metabolism,and reproductive endocrine system in PCOS-IR model rats.EA can increase the expression of HDAC1 and histone H4.In addition,EA can increase the expression of OPA1 and Fis1,decrease the expression of AKT,to reduce oxidative stress and improve IR.
基金supported by the National Natural Science Foundation of China (Grant No. 30600352)the "Top-notch Personnel" Project of Ji-angsu University, the National Basic Research Program of China (Grant No. 2006CB504200)the Open Research Fund Program of the State Key Laboratory of Virology of China (Grant No. 2009008)
文摘HIV-1 co-receptor tropism is central for understanding the transmission and pathogenesis of HIV-1 infection. We performed a genome-wide comparison between the adaptive evolution of R5 and X4 variants from HIV-1 subtypes B and C. The results showed that R5 and X4 variants experienced differential evolutionary patterns and different HIV-1 genes encountered various positive selection pressures, suggesting that complex selection pressures are driving HIV-1 evolution. Compared with other hypervariable regions of Gp120, significantly more positively selected sites were detected in the V3 region of subtype B X4 variants, V2 region of subtype B R5 variants, and V1 and V4 regions of subtype C X4 variants, indicating an association of positive selection with co-receptor recognition/binding. Intriguingly, a significantly higher proportion (33.3% and 55.6%, P【0.05) of positively selected sites were identified in the C3 region than other conserved regions of Gp120 in all the analyzed HIV-1 variants, indicating that the C3 region might be more important to HIV-1 adaptation than previously thought. Approximately half of the positively selected sites identified in the env gene were identical between R5 and X4 variants. There were three common positively selected sites (96, 113 and 281) identified in Gp41 of all X4 and R5 variants from subtypes B and C. These sites might not only suggest a functional importance in viral survival and adaptation, but also imply a potential cross-immunogenicity between HIV-1 R5 and X4 variants, which has important implications for AIDS vaccine development.
文摘Despite the long journey undertaken by mankind to confront the diseases caused by viruses,people were unaware of the existence of viruses until the closing years of the 19th century,when the Russian biologist Dmitri Ivanovsky demonstrated that tobacco plants could be infected by a toxin smaller than bacteria,followed by which the Dutch microbiologist Martinus Beijerinck termed the toxin as a contagium vivum fluidum （soluble living germ） and re-introduced the word ＂virus＂ （derived from the Latin word ＂virus＂ meaning poison; the modern meaning is an ＂agent that causes infectious disease＂ that was first recorded in 1728）.
基金supported by National Insitutes of Health Fogarty International Center grant(1D43TW009532-01)Chinese National Key Technologies R&D Program for the 12th Five-year Plan(2012ZX10001003-003)
文摘Dear Editors,Here, we report a novel HIV-1 intra-circulating recombinant form 01_AE （intra-CRF01_AE： CRF01-1AE/CRF01-6AE） composed of CRF01_AE transmission clusters 1 and 6, which was identified in a heterosexual male from Fujian, with one breakpoint observed in the vif gene. The CRF01_AE region I （HXB2： 868-5184） of the recombinant clustered with CRF01_AE transmission cluster 1, which is mainly circulating among injection drug users （IDUs） and heterosexuals in southern and southwestern China, with the support of a 100% bootstrap value. The CRF01_AE region II （HXB2： 5185-9605） of the recombinant clustered with CRF01_AE transmission cluster 6, which mainly prevails among heterosexuals in southern China, with the support of a 100% bootstrap value. To our knowledge, this is the first detection of a novel HIV-1 intra-CRF01_AE recombinant form （CRF01-1_AE/CRF01-6_ AE） in Fujian, China, which indicates ongoing active HIV-1 transmission among heter- osexuals in this region.
基金supported by the National Natural Science Foundation of China(81371804,81772123 and 81672000)the Thirteen-fifth Key Project(2018ZX10715-005)+1 种基金Beijing Municipal Science and Technology Commission(Z191100006619045)Beijing Hospital authority(DFL20191801 and DFL20191802)。
文摘Chronic human immunodeficiency virus(HIV) infection not only causes a gradual loss of CD4~+T cells but also leads to a disturbance of the T cell receptor(TCR) repertoire. In people living with HIV(PLWH), monitoring TCR repertoire is challenged by the inconsistency of complementarity determining region 3(CDR3) and limited cell numbers in clinical samples. Thus, a quantitative method is necessary for monitoring the TCR repertoire in PLWH.We characterized the TCR V-J pairing profile of na?ve and memory CD4~+T cells in healthy donors, HIV-infected antiretroviral therapy(ART)-na?ve patients and long-term(over5 years) ART-experienced patients by performing TCR sequencing. We developed a V-J index with 18 parameters which were subdivided into five categories(expression coverage, cumulative percentage of the top tenth percentile, diversity, intra-individual similarity and inter-individual similarity). In ART-na?ve patients, 14 of the 18 parameters were significantly altered.Long-term ART recovered ten parameters. The four unrecovered parameters were related to inter-individual similarity.Therefore, these findings indicate that long-term ART could only partially recover TCR V-J pairs and introduce newly impacted V-J pairs. Moreover, these results provide new insights into the V-J pairing of the TCR and into the disturbance of TCR repertoire in HIV infection.
