Intervirology最新文献

{"title":"Prevalence of Hepatitis B Virus Infection Markers among Patients of the Ibn Sina University Hospital Center (Rabat, Morocco).","authors":"Mourad Feindiri, Hakima Kabbaj, Mohammed El Mzibri, Bouchra Belkadi, Najat Bouihat, Abdelkarim Filali-Maltouf, Myriam Seffar","doi":"10.1159/000518618","DOIUrl":"10.1159/000518618","url":null,"abstract":"<p><strong>Introduction: </strong>Viral hepatitis B is a global scourge affecting millions of people worldwide. In Morocco, hepatitis B is considered a public health problem, and available data converge to consider Morocco as a country with intermediate endemicity. In the present study, we have planned to evaluate the HBV prevalence in Morocco on a large scale and to assess the prevalence of different serological markers for better management of this infection in Morocco.</p><p><strong>Methods: </strong>This study was conducted on 18,877 patients referring to the Ibn Sina University Hospital Center of Rabat, Morocco. HBV serological markers including HBsAg, HBsAb, HBeAg, HBeAb, and total HBcAb were assessed by immune-enzymatic assays. The quantification of HBV DNA was performed by real-time PCR.</p><p><strong>Results: </strong>The overall prevalence of positive cases for HBsAg, HBsAb, and total HBcAb was 2.47%, 27.66%, and 21.2%, respectively. From 141 patients with an isolated HBcAb serological profile (HBcAb+/HBsAb-/HBsAg-), HBV DNA was detected in 10 patients, representing a rate of 7.09%. In the present study, up to 95.78% of HBV chronic carriers were negative for HBeAg.</p><p><strong>Conclusion: </strong>This study highlights a higher prevalence of HBsAg in the hospital-based population than the general population reported previously in Morocco and a very low HBV immunization coverage. Of particular interest, detectable HBV DNA levels in isolated HBcAb patients show that exclusive HBsAg screening cannot eliminate the risk of HBV transmission in certain cases. Many efforts are then mandatory to promote serological testing and increase the vaccination rate to limit viral dissemination for better management of this disease in Morocco.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"65 2","pages":"80-86"},"PeriodicalIF":3.2,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9153348/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39413636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acknowledgement to Reviewers","authors":"","doi":"10.1159/000519993","DOIUrl":"https://doi.org/10.1159/000519993","url":null,"abstract":"<br />Intervirology 2021;64:215–216","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"190 1","pages":""},"PeriodicalIF":4.6,"publicationDate":"2021-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138514682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Disinfection effect of hexadecyl pyridinium chloride on SARS-CoV-2 in vitro.","authors":"Keda Chen, Fei Ma, Ying Wang, Xinying Zhuang, Xun-Shan Zhang, Haiyan Mao, Yanjun Zhang","doi":"10.21203/RS.3.RS-458096/V1","DOIUrl":"https://doi.org/10.21203/RS.3.RS-458096/V1","url":null,"abstract":"The novel coronavirus (COVID-19 or 2019-nCoV) is a respiratory virus that can exist in the mouth and saliva of patients and spreads through aerosol dispersion. Therefore, stomatological hospitals and departments have become high-infection-risk environments. Accordingly, oral disinfectants that can effectively inactivate the virus have become a highly active area of research. Hexadecyl pyridinium chloride, povidone-iodine, and other common oral disinfectants are the natural primary choices for stomatological hospitals. Therefore, this study investigated the inhibitory effect of hexadecyl pyridinium chloride on SARS-CoV-2 in vitro. Vero cells infected with SARS-CoV-2 were used to determine the disinfection effect; the CCK-8 method was used to determine cytotoxicity, and viral load was determined by real-time PCR. The results showed that hexadecyl pyridinium chloride has no obvious cytotoxic effect on Vero cells in the concentration range 0.0125-0.05 mg/mL. The in vitro experiments showed that hexadecyl pyridinium chloride significantly inhibits the virus at concentrations of 0.1 mg/mL or above at 2 min of action. Thus, the results provide experimental support for the use of hexadecyl pyridinium chloride in stomatological hospitals.","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"1 1","pages":""},"PeriodicalIF":4.6,"publicationDate":"2021-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42051683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study on the Dynamic Proliferation of JEV in BHK-21 Cells.","authors":"Fuliang Zhang, Jun Luo, Man Teng, Guangxu Xing, Junqing Guo, Yihua Zhang","doi":"10.1159/000510585","DOIUrl":"10.1159/000510585","url":null,"abstract":"<p><strong>Introduction: </strong>Epidemic Japanese encephalitis is one of the most important zoonotic diseases that cause central nervous system damage. The vaccination has become the most effective and economical measure for its control. Hence, real-time monitoring of Japanese encephalitis virus (JEV) proliferation is crucial to optimize virus inoculation, culturing conditions, and virus harvest time.</p><p><strong>Methods: </strong>The proliferation dynamics of JEV in BHK-21 cells was studied by combining the established quantitative PCR method with the conventional TCID50 assay in this study.</p><p><strong>Results: </strong>The proliferation curve determined by the 2 methods has a definite parallel relationship, but the quantitative real-time PCR method (4 h) is faster and more sensitive than the TCID50 method (3-4 days). The determination results of TCID50 showed that the highest viral titer was 105.44 TCID50/0.1 mL and 104.86 TCID50/0.1 mL in cell suspension and culture supernate, respectively, while the virus RNA copies reached the peak at 1.0 × 107.5 copies/µL and 1.0 × 105.6 copies/µL in cell suspension and culture supernate, respectively.</p><p><strong>Conclusion: </strong>The comprehensive analysis showed that the best time for JEV proliferation in BHK-21 cell was 60 h post infection.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":" ","pages":"1-7"},"PeriodicalIF":4.6,"publicationDate":"2021-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38783187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gallid Alphaherpesvirus 2 in the Egyptian Turkeys: Molecular Characterization and Establishment of a Universal System for Phylogenetic Classification.","authors":"Mahmoud Bayoumi,&nbsp;Mohamed El-Saied,&nbsp;Basem Ahmed,&nbsp;Magdy El-Mahdy,&nbsp;Haitham Amer","doi":"10.1159/000515904","DOIUrl":"https://doi.org/10.1159/000515904","url":null,"abstract":"<p><strong>Introduction: </strong>Gallid alphaherpesvirus 2 (GaHV-2) is a highly contagious oncogenic virus that causes Marek's disease in chickens and occasionally in turkeys. Among 100 genes identified in GaHV-2 genome, the Meq gene appears to involve viral virulence, oncogenicity, and genetic diversity. Despite the use of Meq gene sequences in phylogenetic classification of GaHV-2 strains circulating in many countries worldwide, no integrated system exists yet.</p><p><strong>Methods: </strong>Turkeys from 2 commercial Egyptian farms were presented with signs of dullness, dehydration, and emaciation. Samples prepared from the internal organs were examined by histopathology and immunohistochemistry. Pools of the internal organs were analyzed by PCR for identification of GaHV-2, avian leucosis virus, and reticuloendotheliosis virus. The Meq gene of an Egyptian strain was sequenced and analyzed in comparison to 40 reference strains for generation of a universal system for phylogenetic classification of GaHV-2 strains.</p><p><strong>Results: </strong>Gross and histopathological examination revealed grayish-white soft masses in the internal organs characterized by diffuse infiltration of pleomorphic neoplastic cells. All lymphoma cells were identified as T-lymphocytes of CD3+ phenotype. Samples of both farms were only positive for GaHV-2 by PCR. Sequence analysis of the Meq gene has classified the current turkey strain as related to the Egyptian strains identified in chicken in 2012. A universal phylogenetic system for classification of GaHV-2 strains into 4 clusters was proposed. The vaccine strains were all grouped in cluster 2, and most of the classical American strains belonged to cluster 4. Cluster 1 was further divided into 3 subclusters (1.1-1.3).</p><p><strong>Conclusion: </strong>GaHV-2 was identified in turkeys for the first time in Africa and the Middle East. Sequence analysis of the Meq gene of the Egyptian strain along with a wide array of the global strains has enabled the construction of a novel phylogenetic classification system.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 3","pages":"156-164"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000515904","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39009459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glucose Homeostasis Is Dysregulated in Ducks Infected with Duck Hepatitis B Virus.","authors":"Yanlian Tan,&nbsp;Jianxiang Liu,&nbsp;Yingjian Qin,&nbsp;Bin Liang,&nbsp;Yunyan Gu,&nbsp;Lilan Liang,&nbsp;Lili Liu,&nbsp;Yongming Liu,&nbsp;Heling Su","doi":"10.1159/000516766","DOIUrl":"https://doi.org/10.1159/000516766","url":null,"abstract":"<p><strong>Introduction: </strong>The association between hepatitis B virus (HBV) infection and the development of diabetes remains controversial. This study examined the effect of HBV infection on glucose homeostasis using a duck HBV (DHBV) model.</p><p><strong>Methods: </strong>Plasma DHBV DNA was detected by quantitative polymerase chain reaction (PCR). Tissue infection of DHBV was determined by detecting DHBV covalently closed circular DNA (cccDNA) with a method of rolling circle amplification combined with cross-gap PCR, and verified by fluorescence in situ hybridization assay. An intravenous injection glucose tolerance test (GTT) was used to analyze the effect of DHBV infection on glucose tolerance.</p><p><strong>Results: </strong>Of the finally included 97 domestic ducks, 53 (54.6%) were congenitally infected by DHBV. The positive rate of DHBV cccDNA in the liver, kidney, pancreas, and skeletal muscle of the infected ducks was 100, 75.5, 67.9, and 47.2%, respectively. The DHBV-infected ducks had higher blood glucose levels at 15 and 30 min post-load glucose (p < 0.01 and p < 0.001, respectively) in the GTT, much more individuals with greater glucose area under curve (p < 0.01), and a 57% impaired glucose tolerance (IGT) rate, as compared with noninfected controls. In addition, the subgroups of the infected ducks with DHBV cccDNA positive in skeletal muscle maintained the higher blood glucose level up to 2 h post-load glucose during the GTT and had a 76% IGT rate.</p><p><strong>Conclusion: </strong>These results suggest that DHBV intrahepatic and extrahepatic infection impairs glucose tolerance, and thus evidence the association of DHBV infection with the dysregulation of glucose metabolism.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 4","pages":"185-193"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000516766","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39104916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of IL-2 and IFN-γ to EBV Peptides in Stimulated Whole Blood among Multiple Sclerosis Patients and Healthy Individuals.","authors":"Nastaran Rafiee,&nbsp;Mehrdad Ravanshad,&nbsp;Bahador Asadi,&nbsp;Roya Kianfar,&nbsp;Ali Maleki","doi":"10.1159/000517002","DOIUrl":"https://doi.org/10.1159/000517002","url":null,"abstract":"<p><strong>Introduction: </strong>Epstein-Barr virus (EBV), a double-stranded DNA virus, has 2 phases of lytic and latent infection in host cells. After infecting B lymphocytes, EBV becomes persistent in these cells. In healthy individuals, T lymphocytes play a key role in killing EBV-infected B cells. Statistical studies have shown that symptomatic EBV infection increases the risk of MS.</p><p><strong>Methods: </strong>This study intended to measure the immune system's response against the different components of EBV, focusing particularly on T lymphocytes' reaction. Consequently, the mRNA level of IL-2 and IFN-γ, liable for impressing autoimmune diseases and as indicators of T-cell function, was compared in EBNA1- and BRLF1-treated whole blood (WB) cultures of 10 healthy individuals and 10 MS patients using real-time RT-PCR.</p><p><strong>Results: </strong>The analysis of the results demonstrated a significant increased level of IL-2 in MS patients than healthy subjects after exposure to both peptides. Also, the mRNA level of IFN-γ increased in MS patients in EBNA1-treated WB culture.</p><p><strong>Conclusion: </strong>According to the study's results, EBV peptides can reactivate immune cells, especially T lymphocytes, and may indirectly induce inflammation and develop MS; however, it seems that long-time exposure to these peptides has reducing effect on T-cell function and faces the control of infected B lymphocytes with difficulties.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 4","pages":"203-208"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000517002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39111788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Frequency of Cytomegalovirus Viral Load in Iranian Human Immunodeficiency Virus-1-Infected Patients with CD4+ Counts <100 Cells/mm3.","authors":"Mohammad Reza Jabbari,&nbsp;Hoorieh Soleimanjahi,&nbsp;Somayeh Shatizadeh Malekshahi,&nbsp;Mohammad Gholami,&nbsp;Leila Sadeghi,&nbsp;Minoo Mohraz","doi":"10.1159/000514385","DOIUrl":"https://doi.org/10.1159/000514385","url":null,"abstract":"<p><strong>Objectives: </strong>The aim of present work was to assess cytomegalovirus (CMV) viremia in Iranian human immunodeficiency virus (HIV)-1-infected patients with a CD4+ count <100 cells/mm3 and to explore whether CMV DNA loads correlate with CD4+ cell counts or associated retinitis.</p><p><strong>Methods: </strong>This study was conducted at the AIDS research center in Iran on HIV-1-infected patients with CD4+ count <100 cells/mm3, antiretroviral therapy-naive, aged ≥18 years with no previous history of CMV end-organ disease (CMV-EOD).</p><p><strong>Results: </strong>Thirty-nine of 82 patients (47.56%) had detectable CMV viral load ranging from 66 to 485,500 IU/mL. CMV viral load in patients with retinitis ranges from 352 to 2,720 IU/mL, and it was undetectable in 2 patients. No significant associations between CMV viremia and CD4+ cell count was found (p value = 0.31), whereas significant association of CMV viremia in HIV-infected patients with retinitis was found (p < 0.02).</p><p><strong>Conclusions: </strong>We estimated the frequency of CMV viral load infection in Iranian HIV-1-infected patients with a CD4+ cell count <100 mm3/mL in the largest national referral center for HIV-1 infection in Iran. Further research is required on the relevance of CMV viral load in diagnostic and prognostic value of CMV-EOD.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 3","pages":"135-139"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000514385","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25549651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Upregulation of a Novel Long Noncoding RNA AK097647 Promotes Enterovirus 71 Replication and Decreases IFN-λ1 Secretion.","authors":"Min Chu,&nbsp;Bingfei Zhou,&nbsp;Huilin Tu,&nbsp;Min Li,&nbsp;Li Huang,&nbsp;Yuan He,&nbsp;Luo Liu,&nbsp;Song Han,&nbsp;Jun Yin,&nbsp;Biwen Peng,&nbsp;Xiaohua He,&nbsp;Wanhong Liu","doi":"10.1159/000515903","DOIUrl":"https://doi.org/10.1159/000515903","url":null,"abstract":"<p><strong>Background: </strong>Enterovirus 71 (EV71) infects millions of children every year in China and has become a challenge to public health. However, there is no effective treatment for EV71 infection. Long noncoding RNAs (lncRNAs) have been found to play various roles in virus replication and infection.</p><p><strong>Objective: </strong>We aimed to explore the role of a novel long noncoding RNA AK097647 (lncRNA-AK097647) during EV71 infection.</p><p><strong>Methods: </strong>To assess the role of lncRNA-AK097647 during EV71 infection, siRNAs were used to silence lncRNA-K097647 expression. RT-qPCR assay and Western blotting were applied to measure the mRNA and protein levels of EV71 VP1 and the phosphorylation of NF-κB. ELISA was used to detect the level of IFN-λ1 expression.</p><p><strong>Results: </strong>The novel lncRNA-AK097647 was upregulated in human rhabdomyosarcoma cells and the blood of hand, foot, and mouth disease patients infected with EV71, as demonstrated by RT-qPCR. Interestingly, RNAi-mediated knockdown of lncRNA-AK097647 dramatically increased the level of IFN-λ1 expression, resulting in the suppression of EV71 replication. In contrast, overexpression of lncRNA-AK097647 decreased the level of IFN-λ1 expression and resulted in increased EV71 replication. In addition, we found that lncRNA-AK097647 could inhibit the phosphorylation of NF-κB.</p><p><strong>Conclusion: </strong>These results suggest a novel mechanism by which EV71 evades the IFN-mediated host antiviral response by increasing lncRNA-AK097647 expression.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 3","pages":"147-155"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000515903","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38962035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation and Characterization of a Lytic Staphylococcus aureus Phage WV against Staphylococcus aureus Biofilm.","authors":"Yaxian Jiang,&nbsp;Qian Xu,&nbsp;Liming Jiang,&nbsp;Rui Zheng","doi":"10.1159/000515282","DOIUrl":"https://doi.org/10.1159/000515282","url":null,"abstract":"<p><strong>Background: </strong>Staphylococcus aureus is a Gram-positive, pathogenic bacterium that causes a wide range of symptoms in humans and can form biofilm, which is a multicellular community of microorganisms that attaches to nonbiological and biological surfaces.</p><p><strong>Methods: </strong>Here, we aimed to isolate and characterize an S. aureus phage and examine the bactericidal activity alone and in conjunction with streptomycin treatment.</p><p><strong>Results: </strong>We isolated a virulent phage, WV, from a slaughterhouse in Jiangsu, China. This strain belonged to the family Myoviridae and presented a genome size of 141,342 bp. The optimal pH of the preservation buffer was 6-7, optimal growth temperature was 37°C, and optimal multiplicity of infection was 0.01. Phage WV can sterilize most clinical strains of S. aureus that had been isolated from clinical patients in the First People's Hospital of the Yunnan Province. Against low-concentration S. aureus culture, streptomycin demonstrated a greater antibiofilm effect than that of phage WV. By contrast, in high-concentration S. aureus culture, phage WV demonstrated greater antibiofilm effect than that of streptomycin. The use of phage WV and streptomycin together had a substantially greater overall antibiofilm effect than that achieved using either component alone.</p><p><strong>Conclusion: </strong>This study provides strong evidence for the effectiveness of phage application for the reduction of S. aureus biofilm growth and suggests that phages can be considered as a viable alternative to antibiotics in clinical settings.</p>","PeriodicalId":14547,"journal":{"name":"Intervirology","volume":"64 4","pages":"169-177"},"PeriodicalIF":4.6,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000515282","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39154411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}