Antiviral drug resistance increases hepatocellular carcinoma:A prospective decompensated cirrhosis cohort study

AIM:To study the clinical outcome of antiviral therapy in hepatitis B-related decompensated cirrhotic patients.METHODS:Three hundred and twelve patients with decompensated hepatitis B cirrhosis were evaluated in a prospective cohort.With two years of follow-up,198patients in the group receiving antiviral therapy with nucleos(t)ide analogues and 39 patients in the control group without antiviral treatment were analysed.RESULTS:Among the antiviral treatment patients,162had a complete virological response(CVR),and 36 were drug-resistant(DR).The two-year cumulative incidence of hepatocellular carcinoma(HCC)in the DR patients(30.6%)was significantly higher than that in both the CVR patients(4.3%)and the control group(10.3%)(P<0.001).Among the DR patients in particular,the incidence of HCC was 55.6%(5/9)in those who failed rescue therapy,which was extremely high.The rtA181T mutation was closely associated with rescue therapy failure(P=0.006).The Child-Pugh scores of the CVR group were significantly decreased compared with the baseline(8.9±2.3 vs 6.0±1.3,P=0.043).CONCLUSION:This study showed that antiviral drug resistance increased the risk of HCC in decompensated hepatitis B-related cirrhotic patients,especially in those who failed rescue therapy.

Differences in viral kinetics between genotypes 1 and 3 of hepatitis C virus and between cirrhotic and non-cirrhotic patients during antiviral therapy

AIM： TO evaluate the impact of hepatitis C virus （HCV） infection with genotype 1 or 3 and the presence or absence of liver cirrhosis （LC） in the early viral kinetics response to treatment. METHODS： Naive patients （n = 46） treated with interferon-α （IFN-α） and ribavirin and followed up with frequent early HCV-RNA determinations were analysed. Patients were infected with genotype 1 （n = 28, 7 with LC） or 3 （n = 18, 5 with LC）. RESULTS： The first phase decline was larger in genotype 3 patients than in genotype 1 patients （1.72 vs 0.95 log IU/mL, P 〈 0.001）. The second phase slope decline was also larger in genotype 3 patients than in genotype 1 patients （0.87 vs 0.15 log/wk, P 〈 0.001）. Differences were found in both cirrhotic and non-cirrhotic patients. Genotype 1 cirrhotic patients had a slower 2^nd phase slope than non-cirrhotic patients （0.06 vs 0.18 log/wk, P 〈 0,02）. None of genotype 1 cirrhotic patients had a 1^st phase decline larger than 1 log （non-cirrhotic patients： 55%, P 〈 0.02）. A similar trend toward a slower 2nd phase slope was observed in genotype 3 cirrhotic patients but the 1^st phase slope decline was not different. Sustained viral response was higher in genotype 3 patients than in genotype 1 patients ,（72% vs 14%, P 〈 0.001） and in genotype 1 non-cirrhotic patients than in genotype 1 cirrhotic patients （19% vs 0%）. A second phase decline slower than 0.3 log per week was predictive of non-response in all groups. CONCLUSION： Genotype 3 has faster early viral decline than genotype 1. Cirrhosis correlates with a slower 2nd phase decline and possibly with a lower 1^st phase slope decline in genotype 1 patients.

Antiviral effects of a synthetic Aluminium-Magnesium Silicate, on Avian Influenza Virus

Effects a synthetic Aluminium-Magnesium Silicate [AMS] has on Avian Influenza Virus (AIV) were tested. Equal amounts of AIV samples and of the AMS were mixed, kept one hour at room temperature before centrifuging. The supernatants were remeasured and tested for, viral titre, Mean Death Time (MDT) and Mortality Rate of chicken Embryos (EMR). Volumes of the viral samples reduced at rate of 23.4% ± 5.48%. Viral titres reduced significantly (P < 0.01) from HA, 73 ± 32.72 to 1.4 ± 0.43. Also, EMR of infected chicken embryos reduced from 100% to 65%, while MDT of those that died,increased significantly (P < 0.01) from 76 ± 4.38 to 136 ±18.93 hours. When incubation with AMS was repeated on portions of an AIV sample, MDT increased from 64 to 104 hours with the portion incubated once. AIV portions on which incubation with AMS was repeated could not kill chicken embryos.

Putting PLX5622 into perspective:microglia in central nervous system viral infection

Elucidating the exact contribution of microglia to central nervous system(CNS)pathology has historically been extremely challenging.These resident parenchymal myeloid cells are considered to have critical roles as frontline responders during pathogen invasion and CNS perturbation.Thus,understanding the precise temporal kinetics of microglial function is central to the evolution of novel therapeutics for disease intervention and/or resolution(Spiteri et al.,2022a).The development of PLX5622,a colony-stimulating factor 1 receptor(CSF-1R)inhibitor typically formulated into a rodent chow for simple oral administration has facilitated exploration of microglial functions in disease(Spangenberg et al.,2019).

Update on hepatitis B and C virus diagnosis

Viral hepatitis B and C virus(HBV and HCV) are responsible for the most of chronic liver disease worldwide and are transmitted by parenteral route, sexual and vertical transmission. One important measure to reduce the burden of these infections is the diagnosis of acute and chronic cases of HBV and HCV. In order to provide an effective diagnosis and monitoring of antiviral treatment, it is important to choose sensitive, rapid, inexpensive, and robust analytical methods. Primary diagnosis of HBV and HCV infection is made by using serological tests for detecting antigens and antibodies against these viruses. In order to confirm primary diagnosis, to quantify viral load, to determine genotypes and resistance mutants for antiviral treatment, qualitative and quantitative molecular tests are used. In this manuscript, we review the current serological and molecular methods for the diagnosis of hepatitis B and C.

Chronic hepatitis C virus infection:Prevalence of extrahepatic manifestations and association with cryoglobulinemia in Bulgarian patients

AIM: To assess the prevalence of extrahepatic manifestations in Bulgarian patients with chronic hepatitis C virus (HCV) infection and identify the clinical and biological manifestations associated with cryoglobulinemia. METHODS: The medical records of 136 chronically infected HCV patients were reviewed to assess the prevalence of extrahepatic manifestations. Association between cryoglobulin-positivity and other manifestations were identified using χ2 and Fisher’s exact test. Risk factors for the presence of extrahepatic manifestations were assessed by logistic regression analysis. RESULTS: Seventy six percent (104/136) of the patients had at least one extrahepatic manifestation. Clinical manifestations included fatigue (59.6%), kidney impairment (25.0%), type 2 diabetes (22.8%), paresthesia (19.9%), arthralgia (18.4%), palpable purpura (17.6%), lymphadenopathy (16.2%), pulmonary fi brosis (15.4%), thyroid dysfunction (14.7%), Raynaud’s phenomenon (11.8%), B-cell lymphoma (8.8%), sicca syndrome (6.6%), and lichen planus (5.9%). The biological manifestations included cryoglobulinproduction (37.5%), thrombocytopenia (31.6%), and autoantibodies: anti-nuclear (18.4%), anti-smooth muscle (16.9%), anti-neutrophil cytoplasm (13.2%) and anti-cardiolipin (8.8%). All extrahepatic manifestations showed an association with cryoglobulin-positivity, with the exception of thyroid dysfunction, sicca syndrome, and lichen planus. Risks factors for the presence of extrahepatic manifestations (univariate analysis) were: age ≥ 60 years, female gender, virus transmission by blood transfusions, longstanding infection (≥ 20 years), and extensive liver fi brosis. The most signifi cant risks factors (multivariate analysis) were longstanding infection and extensive liver fi brosis. CONCLUSION: We observed a high prevalence of extrahepatic manifestations in patients with chronic HCV infection. Most of these manifestations were associated with impaired lymphoproliferation and cryoglobulin production. Longstanding infection and extensive liver fi brosis were signifi cant risk factors for the presence of extrahepatic manifestations in HCV patients.

Measles Virus IgG Avidity Assay for Use in Identification of Measles Vaccine Failures in Tianjin, China

Objective To identify measles vaccine failures in Tianjin, China using a measles virus Ig G avidity assay.Methods The China Information System for Disease Control and Prevention(CISDCP) was used to collect information about measles cases and blood specimens in Tianjin from 2013 to 2015. Measlesspecific Ig M and Ig G antibodies were detected using Enzyme-Linked Immunosorbent Assay(ELISA).Avidity testing for measles Ig G was performed using a commercial enzyme immunoassay(EIA).Results A total of 284 confirmed measles cases were identified. Of this total, 262(92.25%) were in patients aged ≥ 20 years. High avidity was exhibited in 172(60.56%) cases, while 80(28.17%) cases demonstrated low avidity. High avidity was detected in only 21.43% of cases in patients aged < 1 year.The proportion of high avidity increased with age, and was significantly higher in patients aged 30–39 years at 70.07%(χ~2 = 17.27, P = 0.002). Of the 52 measles cases in patients with a history of vaccinations,41(78.85%) cases showed high avidity, indicating secondary vaccine failures(SVF). In these vaccinations,there was no significant difference(P > 0.05) in clinical severity between high avidity and low avidity cases. However, regardless of vaccination status, clinical severity was significantly lower in high avidity cases(P < 0.001) than in low avidity cases. The percentages of positive measles Ig M results in high avidity and low avidity cases were 66.28% and 91.25%, respectively. Geometric Mean Concentration(GMC) was significantly lower in high avidity cases at 33.73 U/m L, compared to 166.07 U/m L in low avidity cases.Conclusions Low clinical severity and inconclusive Ig M antibody results are more likely in high avidity measles cases. Measles cases were more common in adults. Therefore, a further dose of vaccines should be recommended for 30–39 years in Tianjin.

In vitro study of nonthermal atmospheric pressure plasma in improving the durability of the dentin–adhesive interface with an etch-and-rinse system

In this study, we employed a nonthermal atmospheric pressure plasma(NTAPP) jet to evaluate the effect of plasma treatment on the durability of resin–dentin bonding under a thermocycling challenge. Furthermore, we assessed the degradation resistance of plasma-treated collagen under a sodium hypochlorite(NaClO) challenge. We assessed the beneficial effect of NTAPP treatment on the acid-etched dentin–bonding interface by testing the micro-tensile bond strength and examining the morphology. We found that the immediate bonding strength of the dentin significantly increased after NTAPP treatment. Compared with the control group, NTAPP resulted in a more prominent effect on the bonding durability of the dentin–adhesive interface after treatment for 5 or 10 s. Simultaneously, the mechanical strength of dentin collagen under the NaClO challenge was improved. Our results indicate that, in optimal conditions, NTAPP could be a promising method to protect dentin collagen and to improve the bonding durability between dentin and etch-and-rinse adhesives.

A Pair of Novel Primers for Universal Detection of the NS1 Gene from Various Bluetongue Virus Serotypes

Twenty five serotypes of Bluetongue virus (BTV) have been identified worldwide. Rapid and reliable methods of virus universal detection are essential for fighting against bluetongue (BT). We have therefore developed and evaluated a pair of primers which can detect various serotypes of BTV by RT-PCR. Analysis of the viral protein 7 (VP7) and the non-structural protein (NS1) gene from different serotypes of BTV by DNAstar showed that the 5' end of the NS1 gene is the most conserved region. The primer pairs (P1 and P2) were designed based on the highly conserved region of NS1. The novel primers were evaluated by detecting BTV serotypes 1, 3, 5, 8, 10, 11, 21 and 22. The specificity of the primers was estimated by comparing to gene sequences of viruses published in GenBank, and further assessed by detecting BTV serotype 1-12 and Epizootic hemorrhagic disease virus (EHDV) serotype 1-4. The sensitivity and repeatability of PCR with the novel primers were evaluated by successfully detecting the recombinant plasmid pGEM-T121 containing the diagnosed nucleotide sequence. Our results suggest that these unique primers can be used in high throughout and universal detection of the NS1 gene from various BTV serotypes.

Expression of IL-1β and TNF-α in MCMV Myocarditis and Its Role

In order to study the expression of IL-1β and TNF-α in the myocardium of MCMV myocarditis and their role in the myocardial damages, 60 BALB/C mice of 4 weeks were randomly divided into two groups: 36 were injected intraperitoneally with MCMV and 24 served as control group. Immunohistochemistry was used to detect IL-1β and TNF-α expression in the myocardium, and myocardial lesions were observed histopathologically. Histopathological study on the myocardium from infected mice revealed focal or diffuse lesions characterized by inflammatory cells and degeneration or necrosis of myocytes. The myocardial lesion score showed the degree of inflammatory cell infiltration was slight in MCMV myocarditis.The positive staining signals for IL-1β and TNF-α proteins which mainly located in the infiltrating inflammatory cells and degenerative or necrotic myocytes were markedly detectable whereas there were no positive findings in the myocardium of control mice. IL-1β and TNF-α was expressed in the myocardium of viral myocarditis murine model induced by MCMV. IL-1β and TNF-α may play an important role in the pathogenesis of viral myocarditis.

Predictive factors for survival and score application in liver retransplantation for hepatitis C recurrence

AIM: To identify risk factors associated with survival in patients retransplanted for hepatitis C virus(HCV) recurrence and to apply a survival score to this population.METHODS: We retrospectively identified 108 patients retransplanted for HCV recurrence in eight European liver transplantation centers(seven in France, one in Spain). Data collection comprised clinical and laboratory variables, including virological and antiviral treatment data. We then analyzed the factors associated with survival in this population. A recently published score that predicts survival in retransplantation in patients with hepatitis C was applied. Because there are currently no uniform recommendations regarding selection of the best candidates for retransplantation in this setting, we also described the clinical characteristics of 164 patients not retransplanted, with F3, F4, or fibrosing cholestatic hepatitis(FCH) post-first graft presenting with hepatic decompensation. RESULTS: Overall retransplantation patient survival rates were 55%, 47%, and 43% at 3, 5, and 10 years, respectively. Patients who were retransplanted for advanced cirrhosis had survival rates of 59%, 52%, and 49% at 3, 5, and 10 years, while those retransplanted for FCH had survival rates of 34%, 29%, and 11%, respectively. Under multivariate analysis, and adjusting for the center effect and the occurrence of FCH, factors associated with better survival after retransplantation were: negative HCV viremia before retransplantation, antiviral therapy after retransplantation, non-genotype 1, a Model for End-stage Liver Disease(MELD) score < 25 when replaced on the waiting list, and a retransplantation donor age < 60 years. Although the numbers were small, in the context of the new antivirals era, we showed that outcomes in patientswho underwent retransplantation with undetectable HCV viremia did not depend on donor age and MELD score. The Andrés score was applied to 102 patients for whom all score variables were available, producing a mean score of 43.4(SD = 6.6). Survival rates after the date of the first decompensation post-first liver transplantation(LT1) in the liver retransplantation(re LT) group(94 patients decompensated) at 3, 5, and 10 years were 62%, 59%, and 51%, respectively, among 78 retransplanted individuals with advanced cirrhosis, and 42%, 32%, and 16% among 16 retransplanted individuals with FCH. In the non-re LT group with hepatic decompensation, survival rates were 27%, 18%, and 9% at 3, 5, and 10 years, respectively(P < 0.0001). Compared with non-retransplanted patients, retransplanted patients were younger at LT1(mean age 48 ± 8 years compared to 53 ± 9 years in the no re LT group, P < 0.0001), less likely to have human immunodeficiency virus(HIV) co-infection(4% vs 14% among no re LT patients, P = 0.005), more likely to have received corticosteroid bolus therapy after LT1(25% in re LT vs 12% in the no re LT group, P = 0.01), and more likely to have presented with sustained virological response(SVR) after the first transplantation(20% in the re LT group vs 7% in the no re LT group, P = 0.028).CONCLUSION: Antiviral therapy before and after retransplantation had a substantial impact on survival in the context of retransplantation for HCV recurrence, and with the new direct-acting antivirals now available, outcomes should be even better in the future.

Dried blood spot sampling as an alternative for the improvement of hepatitis B and C diagnosis in key populations

BACKGROUND To achieve the elimination of hepatitis B and C,there is an urgent need to develop alternative strategies to increase the access of diagnosis,particularly among key populations such as people living with human immunodeficiency virus(HIV),individuals with coagulopathies and chronic kidney disease(CKD)patients.AIM To evaluate the use of dried blood spot(DBS)in the detection of hepatitis B virus(HBV)and hepatitis C virus(HCV)markers.METHODS A total of 430 individuals comprised of people living with HIV,coagulopathies and CKD provided paired serum and DBS samples.HBsAg,anti-HBc and anti-HCV were tested in those samples using a commercial electrochemiluminescence.Demographic and selected behavioral variables were evaluated to assess possible association with HBV and HCV positivity.RESULTS Using DBS,HBsAg prevalence varied from 3.9%to 22.1%,anti-HBc rates varied from 25.5%to 45.6%and anti-HCV positivity ranged from 15.9%to 41.2%in key populations.Specificities of HBV and HCV tests using DBS varied from 88.9%to 100%.The HBsAg assay demonstrated the best performance in CKD and coagulopathy individuals and the anti-HCV test had a sensitivity and specificity of 100%in people living with HIV.Accuracy of HBV and HCV detection in DBS varied from 90.2%to 100%.In the CKD group,HBsAg positivity was associated with infrequent use of condoms,and anti-HBc positivity was associated with sharing nail cutters/razors/toothbrushes.Anti-HCV reactivity was positively associated with a history of transplantation and length of time using hemodialysis in both specimens.In people living with HIV,only the male gender was associated with anti-HBc positivity in serum and DBS.CONCLUSION DBS with electrochemiluminescence are useful tools for the diagnosis and prevalence studies of hepatitis B and C among key populations and may increase the opportunity to foster prevention and treatment.

MicroRNAs as Important Players in Host-hepatitis B Virus Interactions

Hepatitis B virus (HBV) infection,a major public health problem,causes acute and chronic hepatitis that is often complicated by liver cirrhosis and hepatocellular carcinoma.The pathogenic mechanisms of HBV-related liver disease are not well understood,and the current licensed therapies are not effective in permanently clearing virus from the circulation.In recent years,the role of micro-ribonucleic acids (miRNAs) in HBV infection has attracted great interest.Cellular miRNAs can influence HBV replication directly by binding to HBV transcripts and indirectly by targeting cellular factors relevant to the HBV life cycle.They are also involved in the regulation of cellular genes and signaling pathways that have critical roles in HBV pathogenesis.HBV infection,in turn,can trigger changes in cellular miRNA expression that are associated with distinctive miRNA expression profiles depending on the phase of liver disease.These alterations in miRNA expression have been linked to disease progression and hepatocarcinogenesis.We provide here an up to date review regarding the field of miRNAs and HBV interplay and highlight the potential utility of miRNAs as diagnostic biomarkers and therapeutic targets for the management of HBV-related liver disease.

The immune tolerant phase of chronic HBV infection： new perspectives on an old concept

Chronic hepatitis B virus （HBV） infection progresses through distinct disease phases that are strongly associated with patient age. The so-called immune tolerant （IT） phase represents the classical early phase of infection; it is associated with high levels of HBV replication and lack of clinical signs of liver Inflammation. Whether this phase of HBV infection is also associated with immunological features of ＂tolerance＇ has recently been challenged. Here, we review the data that dispute this concept of immune tolerance and then propose an alternative interpretation of the immunopathological events that take place during this early phase of CHB infection.

Supplementing Conventional Treatment with Pycnogenol(R) May Improve Hepatitis C Virus-Associated Type 2 Diabetes:A Mini Review

Hepatitis C virus (HCV) infection and type 2 diabetes mellitus (T2DM) present a significant health burden,with increasing complications and mortality rates worldwide.Pycnogenol(R) (PYC),a natural product,possesses antidiabetic and antiviral properties that may improve HCV-associated T2DM.In this review,we present previously published data on the effectiveness of PYC against HCV replication and T2DM.We believe that supplementing conventional treatment with PYC may improve the current HCV therapy,attenuate HCV-associated T2DM,and reduce the risk of complications such as cirrhosis or hepatocellular carcinoma and cardiovascular disease.

Construction of coxsackievirus B5 viruses with luciferase reporters and their applications in vitro and in vivo

Coxsackievirus belongs to the Picornaviridae family and is one of the major pathogens that cause hand,foot and mouth disease(HFMD)in infants and children with potential serious complications and even deaths.The pathogenesis of this virus is not fully elucidated and no vaccine or antiviral drug has been approved.In this study,a full-length infectious cDNA clone of coxsackievirus B5 virus was assembled and the recombinant virus displayed similar growth kinetics and ability to cause cytopathic effects as the parental virus.Luciferase reporter was then incorporated to generate both full-length and subgenomic replicon(SGR)reporter viruses.The full-length reporter virus is suitable for high-throughput antiviral screening,while the SGR is a useful tool to study viral-host interactions.More importantly,the full-length reporter virus has also been shown to infect the suckling mouse model and the reporter gene could be detected using an in vivo imaging system,thus providing a powerful tool to track viruses in vivo.In summary,we have generated coxsackievirus B5 reporter viruses and provided unique tools for studying virus-host interactions in vitro and in vivo as well as for high-throughput screenings(HTS)to identify novel antivirals.

Effect of Peroxisome Proliferator-Activated Receptor-γ Coactivator-1 Alpha Variants on Spontaneous Clearance and Fibrosis Progression during Hepatitis C Virus Infection in Moroccan Patients

Hepatitis C virus(HCV)is still one of the main causes of liver disease worldwide.Metabolic disorders,including nonalcoholic fatty liver disease(NAFLD),induced by HCV have been shown to accelerate the progression of fibrosis to cirrhosis and to increase the risk of hepatocellular carcinoma.An optimal peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PPARGC1A)activity is crucial to prevent NAFLD installation.The present study aims to investigate the associations between two common PPARGC1A polymorphisms(rs8192678 and rs12640088)and the outcomes of HCV infection in a North African context.A series of 592 consecutive Moroccan subjects,including 292 patients with chronic hepatitis C(CHC),100 resolvers and 200 healthy controls were genotyped using a TaqMan allelic discrimination assay.PPARGC1A variations at rs8192678 and rs12640088 were not associated with spontaneous clearance of HCV infection(adjusted ORs=0.76 and 0.79 respectively,P[0.05,for both).Furthermore,multivariable logistic regression analysis showed that both SNPs were not associated with fibrosis progression(OR=0.71;95%CI 0.20–2.49;P=0.739;OR=1.28;95%CI 0.25–6.54;P=0.512,respectively).We conclude that,in the genetic context of South Mediterranean patients,rs8192678 and rs12640088 polymorphisms of PPARGC1 A are neither associated with spontaneous clearance nor with disease progression in individuals infected with HCV.

Directed shift of vaginal flora after topical application of sucrose gel in a phase Ⅲ clinical trial： a novel treatment for bacterial vaginosis

Background Bacterial vaginosis （BV） is one of the most common infectious diseases among sexually active women and is associated with the increased acquisition of a variety of sexually transmitted diseases.This study aimed to compare the efficacy of a non-antibiotic sucrose gel against an antibiotic metronidazole gel for the treatment of BV.Methods A randomized, double-blinded, multi-center, parallel-group, placebo-controlled phase Ⅲ clinical trial was conducted at eight hospitals in China.A total of 560 subjects with clinically diagnosed BV were randomly assigned into three groups for vaginally receiving sucrose, metronidazole, and placebo gels, respectively, twice daily for five consecutive days.The efficacy of therapeutic cure, defined as an achievement of both microbiologic cure （a Nugent score of 3 or less） and clinical cure （a resolution of the clinical findings from the baseline visit）, was evaluated at the 1st and 2nd test-of-cure （TOC） visits at 7-10 and 21-35 days after the start of treatment, respectively.Results Therapeutic cure rates for sucrose, metronidazole, and placebo gel groups were 83.13%, 71.30% and 0.92%,at the 1st TOC, and 61.04%, 66.67% and 7.34%, at the 2nd TOC, respectively.While there was no significant difference between the sucrose and metronidazole gel groups at the 2nd TOC （P=0.305）, and sucrose gel was more effective than metronidazole gel at the 1st TOC （P=0.009）.Conclusion These findings suggest that sucrose gel restores normal vaginal flora more rapidly than metronidazole gel and can be used as a novel treatment for BV.