Subtype 1b Research Articles

Development and validation of a simple and rapid method for hepatitis�C virus genotyping based on one‑step RT‑qPCR

Hepatitis C virus (HCV) infections caused by different subtypes require different treatments; therefore, rapid and cost-effective genotyping methods for the diagnosis of HCV are greatly needed. In the present study, a new method to diagnose HCV subtypes that depends on a one-step quantitative reverse transcription PCR (RT-qPCR) and TaqMan fluorescence probe technique is described. Five pairs of primers and five probes were designed, which were able to detect five genotypes in three reaction tubes. One reaction was used to detect the 1b subtype, one was used to detect the 2a and 6a subtypes, and the other was used to detect the 3a and 3b subtypes. Rigorous performance validation was implemented for five aspects: Precision, sensitivity, accuracy, specificity and anti-interference. The HCV subtype that infected 289 patients was evaluated in the present study via RT-qPCR and verified by sequencing. The results revealed that the 1b subtype accounted for 45% of infections, the 2a subtype accounted for 9% of infections, the 3a subtype accounted for 13% of infections, the 3b subtype accounted for 18% of infections, and the 6a subtype accounted for 15% of infections. The analytical sensitivity for the detection of each of the five HCV subtypes was 1,000 IU/ml. The new method performed well in the performance validation mentioned above, indicating its effectiveness as a HCV genotyping method. RT-qPCR has mitigated some of the former challenges of existing HCV genotyping methods, including the time commitment, expense, and inaccuracy of such methods. The performance validation of this new method showed that RT-qPCR is reliable enough to be widely applied in China for HCV genotyping.

Spontaneous coronary artery dissection in young female acute coronary syndrome patients: a single-centre retrospective cohort study

Background Spontaneous coronary artery dissection (SCAD) is an unusual cause of acute coronary syndrome (ACS), rapidly gaining recognition over the last decade. SCAD occurs predominantly in young, otherwise healthy women and coronary angiogram often lacks typical (atherosclerotic) features. Therefore, SCAD remains notably underdiagnosed. As optimal treatment strategy differs greatly from atherosclerotic ACS, early (differential) diagnosis is crucial. Purpose/Methods In this paper, all coronary angiograms performed for ACS in women up to 50 years of age were retrospectively reviewed by three independent interventional cardiologists. Results The obtained insights are comparable to recent literature. SCAD incidence was 26% in this cohort. Left anterior descending coronary artery was the main affected vessel with SCAD subtype 2B as predominant angiographic presentation. Correct diagnosis during index procedure was poor with only 33% accuracy. Nevertheless, a favourable trend over time was noted. Percutaneous coronary intervention success was 56%, as in 44% of patients initial stenting was complicated by progressive dissection. Overall, outcome was excellent with no reported fatalities. Conclusion SCAD remains an underdiagnosed subtype of ACS and the importance of increasing awareness amongst (interventional) cardiologists needs to be emphasised.

Dysregulated Prefrontal Cortex Inhibition in Prepubescent and Adolescent Fragile X Mouse Model.

Changes in excitation and inhibition are associated with the pathobiology of neurodevelopmental disorders of intellectual disability and autism and are widely described in Fragile X syndrome (FXS). In the prefrontal cortex (PFC), essential for cognitive processing, excitatory connectivity and plasticity are found altered in the FXS mouse model, however, little is known about the state of inhibition. To that end, we investigated GABAergic signaling in the Fragile X Mental Retardation 1 (FMR1) knock out (Fmr1-KO) mouse medial PFC (mPFC). We report changes at the molecular, and functional levels of inhibition at three (prepubescence) and six (adolescence) postnatal weeks. Functional changes were most prominent during early postnatal development, resulting in stronger inhibition, through increased synaptic inhibitory drive and amplitude, and reduction of inhibitory short-term synaptic depression. Noise analysis of prepubescent post-synaptic currents demonstrated an increased number of receptors opening during peak current in Fmr1-KO inhibitory synapses. During adolescence amplitudes and plasticity changes normalized, however, the inhibitory drive was now reduced in Fmr1-KO, while synaptic kinetics were prolonged. Finally, adolescent GABAA receptor subunit α2 and GABAB receptor subtype B1 expression levels were different in Fmr1-KOs than WT littermate controls. Together these results extend the degree of synaptic GABAergic alterations in FXS, now to the mPFC of Fmr1-KO mice, a behaviourally relevant brain region in neurodevelopmental disorder pathology.

Prevalence of autoimmune diseases in thymic epithelial tumors (TET) insights from RYTHMIC.

9073 Background: TET are associated with autoimmune disorders (AID) in up to 30% of patients (pts). However, there have been wide variations in the reported prevalence of AID in TET pts in small single-center series. RYTHMIC (Réseau tumeurs THYMiques et Cancer) is a French network mandated to systematically discuss every case of TET. We aimed to describe the prevalence of AID in a large French population. Methods: RYTHMIC database, hosted by IFCT (Intergroupe Francophone de Cancérologie Thoracique), prospectively includes all consecutive pts with a diagnosis of TET discussed in French national or regional tumor boards. We analyzed epidemiologic, clinical and pathological characteristics of pts with TET’s related AID. Results: From January 2012 to December 2019, 2909 pts were included in the database. The mean age at diagnosis of TET was 54 and 52% were male. In the overall population, Masaoka Koga stages were well balanced with 12.6% (n = 187) stage I, 8.8% (n = 131) stage IIa, 8.4% (n = 124) stage IIb, 11.1% (n = 164) stage III and 8.5% (n = 125) stage IV. There were 364 (12.5%) events of AID in 302 pts. 62 pts (17%) had more than 1 AID. Among the events, 236 were myasthenia gravis (MG) (64.8%), 19 Hypo-gammaglobulinemia syndrome (5.2%), 15 pure red cell aplasia (4.1%), 18 thyroiditis (4.9%) and 16 systemic erythematous lupus (4.4%). Diagnosis of AID was mostly done at tumor diagnosis (n = 239, 65.7%) but some patient had AID diagnosed before diagnosis (n = 67, 18.4%) or during follow up (n = 32, 8.8%). Among pts presenting AID, B2 was the most common subtype (n = 133, 36.5%). The incidence of AID per subtype was as follow: A (n = 10/81, 12.3%), AB (n = 48/225, 21.3%), B1 (n = 35/130, 26.9%), B2 (n = 133/295, 45.0%), B3 (n = 46/113, 40.7%), thymic carcinoma (n = 16/275, 5.8%). Conclusions: The prevalence of AID in pts with TET was 12.5%, > 40% in B2 and B3 subtypes. Diagnosis of AID can be delayed compared to the diagnosis of TET. Immunotherapy indication should be carefully assessed in pts with TET other than thymic carcinoma.

Canine parvovirus 2b in fecal samples of asymptomatic free-living South American coatis (Nasua nasua, Linnaeus, 1766).

Canine parvovirus type 2 (CPV-2) is classified into three subtypes (CPV-2a, CPV-2b, and CPV-2c) and is the main cause of enteritis and myocarditis in young domestic and wild animals. This study aimed to evaluate the presence of CPV-2 in the feces of asymptomatic free-living coatis from Garden Forest Reserve, Palmital city, SP, Brazil. Fecal samples from 21 coatis (both sexes, different ages, and different aspects of feces) were collected in August 2014 and March 2015. The nucleic acid extracted was submitted to a polymerase chain reaction (PCR) assay to amplify a fragment of the VP2 gene of CPV-2. Eight (38%) fecal samples were positive in the PCR assay and were confirmed by sequencing. The 7 nucleotide (nt) sequences analyzed showed 100% nt identity with the prototype strain of CPV-2b (CPV-39 strain). The analysis of the deduced amino acid (aa) sequence revealed the presence of the GAT codon (aa D-Asp) at position 426 of the VP2 viral protein (subtype 2b). This study describes for the first time the identification of CPV-2b in asymptomatic free-living coatis (Nasua nasua) and suggests that coatis are susceptible to Carnivore protoparvovirus 1 infection and are important as a reservoir and an asymptomatic carrier to other wild and domestic animal species.

Effects of a Novel Psychomotor Stabilizer, IRL790, on Biochemical Measures of Synaptic Markers and Neurotransmission.

The novel small-molecule psychomotor stabilizer, IRL790, is currently in clinical trial for treatment of levodopa-induced dyskinesia and psychosis in patients with Parkinson disease. Here, we used naïve mice to investigate the effects of acute systemic administration of IRL790 on protein levels and phosphorylation states of proteins relevant for synaptic plasticity and transmission. IRL790 increased pro-brain-derived neurotrophic factor protein levels and phosphorylation at Ser1303 of the N-methyl-D-aspartate (NMDA) subtype 2B glutamate receptor (NR2B) in prefrontal cortex. IRL790 also increased the phosphorylation states at Ser19, Ser31, and Ser40, respectively, of tyrosine hydroxylase in striatum. IRL790 reduced protein levels of the NR2B receptor in striatum but not in prefrontal cortex. Taken together, we report that systemically administered IRL790 rapidly elicits changes in protein level and phosphorylation state of proteins associated with a beneficial effect on synaptic markers and neurotransmission. SIGNIFICANCE STATEMENT: The novel small-molecule psychomotor stabilizer, IRL790, is currently in clinical trial for treatment of levodopa-induced dyskinesia and psychosis in patients with Parkinson disease. In this study, we report that systemically administered IRL790 rapidly elicits changes in protein level and phosphorylation state of proteins associated with a beneficial effect on synaptic markers and neurotransmission.

Treatment options for proximal humeral fractures in the older adults and their implication on personal independence.

No consensus exists on the optimal treatment of proximal humeral fractures (PHFx). Uncertainty about surgical treatment in the older adults using locking plates (e.g., PHILOS) has emerged, due to a high number of complications. This study aimed to assess the impact of non-operative versus operative treatment of a PHFx on the level of self-dependence in our older population. We included patients aged over 65years with some level of self-dependence, treated at our hospital between 5/2011 and 4/2013 for isolated PHFx of AO subtypes A2, A3, and B1 for which either non-operative or surgical treatment using a PHILOS plate had been applied. The patients were questioned, examined, or interviewed via phone; AO fracture patterns and treatment were documented as well as level of self-dependence, complications, constant score (CSM), subjective shoulder value (SSV), quality of life (EQ-5D), and shoulder pain and disability index (SPADI). Patients with PHFx of AO subtypes A2, A3, or B1 that were either treated non-operative (n = 50) or operative by insertion of the PHILOS plate (n = 63) were included. Operative-treated patients were 3.3 times as likely to lose some level of independence (95% CI 0.39-28, p = 0.271). Shoulder motion, strength, and functional outcomes tended to be lower in operative-treated patients, with adjusted differences of, - 11 CMS points (95% CI - 23 to 2), - 9 SPADI points (95% CI - 18 to 0), and - 6% in SSV (95% CI - 17 to 5). Quality-of-life EQ-5D utility index was similar in both groups (mean - 0.04; 95% CI - 0.18 to 0.10). In our study population, non-operatively treated older adults with an AO type A2, A3, B1 fracture of the proximal humerus tended to have a high chance to return to their premorbid level of independence, compared to patients treated with a locking plate. A change in the treatment algorithm for these PHFx may be carefully considered and further investigated in clinical practice.

Immune response to bovine viral diarrhea virus (BVDV) vaccines detecting antibodies to BVDV subtypes 1a, 1b, 2a, and 2c

Bovine viral diarrhea virus (BVDV) represents a major cattle disease with multiple forms including fetal infections resulting in persistently infected (PI) cattle. The objectives of this study were to investigate the immune response to six vaccines, five modified live viral (MLV) and one killed vaccine containing BVDV immunogens as measured by antibodies to BVDV1a, BVDV1b, BVDV2a, and BVDV2c. The predominant BVDV subgenotype in the U.S. is BVDV1b compared to BVDV1a and BVDV2a. There are MLV and killed BVDV vaccines containing BVDV1a and BVDV2a marketed in the U.S. A prior study evaluated immune response to vaccination with BVDV1a and BVDV2a inducing virus neutralizing antibody titers. BVDV1b titers 128 or higher at time of exposure to BVDV1b PI cattle protected heifers against fetal infection. Calves received two doses and postweaning serums were collected and assayed for BVDV antibodies. Antibody titers were expressed as geometric mean averages. Percentages were expressed as proportions of animals within three antibody levels, including targeted level 128 or greater. There were statistical differences among vaccines in each study, particularly to BVDV1a, BVDV1b, and BVDV2a. MLV vaccines containing Singer strain induced higher levels to BVDV1a and BVDV1b than NADL vaccine in all three studies. Two vaccines, both MLV, Vaccine 1 and Vaccine 6 containing Singer strain induced higher proportion of 128 or higher BVDV1b titers than vaccine with NADL. Antibody levels to BVDV2a and BVDV2c were dependent on BVDV2a vaccine strain. This study indicates strain in BVDV vaccines reflects differences in immune response to different BVDV subgenotypes, particularly BVDV1b and BVDV2c.

Occult HCV and occult HBV coinfection in Iranian human immunodeficiency virus-infected individuals.

The presence of hepatitis C virus (HCV) genome in liver biopsy or peripheral blood mononuclear cell (PBMC) specimens in the absence of detectable HCV-RNA in plasma of the people with or without anti-HCV antibodies has defined as occult HCV infection (OCI), whereas occult hepatitis B virus infection (OBI) is detection of hepatitis B virus (HBV) genome in the absence of traceable hepatitis B surface antigen in the plasma samples of patients. The purpose of this study is to determine the presence of OBI and OCI in human immunodeficiency virus (HIV)-infected individuals. In this cross-sectional research, 190 Iranian HIV-infected individuals were enrolled from September 2015 to February 2019. All participants were tested regarding various serological markers for HCV and HBV infections. Viral RNA and DNA were extracted from plasma and PBMC specimens, and the presence of HCV-RNA in plasma and PBMC samples was tested using reverse transcriptase-nested polymerase chain reaction (PCR), HBV viral load was determined in plasma samples using COBAS TaqMan 48 Kit, and also the presence of the HBV-DNA in PBMC samples was tested by real-time PCR. In this study, the prevalence of OBI and OCI in HIV-infected individuals was 3.1% and 11.4%, respectively. The genotypes of HCV in the patients with OCI were as follows: 57.1% were infected with subtype 3a, 35.7% were infected with subtype 1a, and 7.1% was infected with subtype 1b. It is noteworthy that in this study, two patients (1.1%) had OCI/OBI coinfections. The present study revealed that 1.1% of Iranian HIV-infected individuals had OBI and OCI at the same time. Therefore, it seems that designing prospective surveys to determine the presence of this coinfection in HIV-infected individuals is informative.

Perfil Epidemiológico dos portadores de Hepatite C atendidos nos Serviços de Referência no Estado do Piauí

A hepatite C é um problema de saúde global e constitui uma das principais causas de doenças hepáticas crônicas em todos os continentes. O objetivo do artigo é apresentar o perfil epidemiológico da hepatite C no Estado do Piauí. Trata-se de um estudo retrospectivo, transversal e descritivo com abordagem quantitativa e avaliativa.Foram coletados no SINAN, livros de registros e prontuários, referentes aos pacientes diagnosticados com o Vírus da Hepatite C que foram atendidos no intervalo de 2014 a 2016 no estado do Piauí. Os resultados obtidos mostraram prevalência do sexo masculino, todavia, a baixa escolaridade encontra-se equivalente para ambos os sexos. Os municípios de maiores notificações foram: Teresina (capital do Estado), seguido de Parnaíba, Piripiri e Picos. Foi possível identificar os genótipos e seus subtipos, sendo o tipo 1 (com subtipos 1a e 1b) mais notificados no Estado, seguido do genótipo tipo 3. As vias de transmissão encontradas foram: via parenteral, transfusão sanguínea e sexual. Quanto ao tratamento observa-se que em 2014, a terapia dupla foi realizada com as drogas Peginterferon alfa e Ribavirina; em 2015 e 2016, a terapêutica foi atualizada para Sufosbuvir, Simeprevir e Daclastavir, no entanto, os tratamentos variavam de acordo com o genótipo identificado. Constata-se que entre 2014 e 2016 houve uma discrepância entre as notificações dos casos notificados no SINAN e o registro do número de pacientes na dispensação dos medicamentos, onde este mostrou-se com maiores registos. Dados que devem chamar atenção do Estado para a descentralização das notificações.

Development and Validation of a Procedure for Authenticity Verification of Modified Lipopolysaccharides of Shigella flexneri Subtypes 1b, 2a, 3a, 6, and Y by 13C NMR Spectroscopy

Clinically applicable lipopolysaccharides (LPSs) from Shigella flexneri, lipid A of which contains at most three higher fatty-acid residues, are promising candidates for creating vaccines against it. NMR studies of modified LPS are practically unreported. Most publications in this field are devoted to studying the polysaccharide structure of native LPS, i.e., O-specific polysaccharide (OPS) that can be isolated after mild hydrolysis of LPS and separation of lipid A. The present article describes the development of an NMR procedure for authenticity verification of modified LPS, i.e., vaccine polysaccharide components. Characteristic 13C resonances in the region of anomeric C atoms are identified. The confidence intervals of their chemical shifts are determined. The procedure is demonstrated to be specific for modified LPS subtypes 1b, 2a, 3a, Y, and 6 from S. flexneri.

Ultrasensitive Detection of Hepatitis C Virus DNA Subtypes Based on Cucurbituril and Graphene Oxide Nano-composite

Infectious of hepatitis C viruses(HCVs) lead to hepatic fibrosis, cirrhosis even hepatoma. Developing rapid and sensitive diagnostic method for HCV is of great importance. Based on the host-and-guest interaction between cucurbit[7]uril(CB[7]) and methylene blue(MB), a CB[7]-graphene nano-composite(CB[7]-N3-GO) is raised for the electrochemical detection of HCV DNA. The method is able to linearly detect the HCV nucleic acid in the range of 0.2–10 nmol/L with detection limit as low as 160.4 pmol/L. The proposed detection strategy is able to discriminate the 1b and 6k subtypes of HCV and has a prospective potential in the blood screen for HCV in clinical diagnosis.

Phylogenetic Analysis of Belgian Small Ruminant Lentiviruses Supports Cross Species Virus Transmission and Identifies New Subtype B5 Strains.

Small ruminant lentiviruses (SRLV) are a group of highly divergent viruses responsible for global and fatal infections in sheep and goats. Since the current phylogenetic classification of these viruses was proposed in 2004, it nowadays consists out of 5 genotypes and 28 subtypes. In support of our national SRLV control program, we performed the genetic characterization of SRLV strains circulating in the Belgian sheep and goat population. Fourteen sheep and 9 goat strains were sequenced in the gag-pol and pol regions using the method described by Shah. Most SRLV strains from sheep and goats belonged to prototype A1 and B1 subtypes, respectively. We, however, also found indications for cross-species transmission of SRLV strains between sheep and goats and vice versa, and identified a new subtype designated as B5. An in-depth analysis of the current SRLV phylogeny revealed that many subtypes have been defined over the years based on limited sequence information. To keep phylogeny as a useful tool, we advocate to apply more rigorous sequencing standards to ensure the correct classification of current and new emerging strains. The genetic characterization of Belgian SRLV strains will help in the development of appropriate diagnostic tools to assist the national control program.

Comparative Antennal Morphology of Agriotes (Coleoptera: Elateridae), with Special Reference to the Typology and Possible Functions of Sensilla.

Species of the click-beetle genus Agriotes Eschscholtz are economically important crop pests distributed mainly in the Northern Hemisphere. They can inflict considerable damage on various field crops. Therefore, the detection, monitoring, and control of Agriotes include the adult trapping using species-specific sex pheromones, which is a critical component of pest research. To obtain a better understanding of the detailed antennal morphology as background information for subsequent chemical ecology research, we conducted a scanning electron microscopy study of the antennal sensilla of both sexes in 10 European Agriotes species. We identified 16 different sensilla in Agriotes, belonging to six main types: sensilla chaetica (subtypes C1 and C2), sensilla trichodea, sensilla basiconica (subtypes B1–B9), dome-shaped sensilla (subtypes D1 and D2), sensilla campaniformia, and Böhm sensilla. We discuss their possible functions and compare the sensilla of Agriotes with those of other Elateridae in order to consolidate the sensillum nomenclature in this family. Additionally, our study reveals the remarkable interspecific variability in sensillar equipment of Agriotes and identifies several characters of potential importance for future use in systematic studies. The present study provides a strong preliminary framework for subsequent research on the antennal morphology of this crop pest on a wider scale.

The Reliability of the AOSpine Thoracolumbar Classification System in Children: Results of a Multicenter Study.

The purpose of this study was to determine whether the new AOSpine thoracolumbar spine injury classification system is reliable and reproducible when applied to the pediatric population. Nine POSNA (Pediatric Orthopaedic Society of North America) member surgeons were sent educational videos and schematic papers describing the AOSpine thoracolumbar spine injury classification system. The material also contained magnetic resonance imaging and computed tomography imaging of 25 pediatric patients with thoracolumbar spine injuries organized into cases to review and classify. The evaluators classified injuries into 3 primary categories: A, B, and C. Interobserver reliability was assessed for the initial reading by Fleiss kappa coefficient (kF) along with 95% confidence interval (CI). For A and B type injuries, subclassification was conducted including A0 to A4 and B1 to B2 subtypes. Interobserver reliability across subclasses was assessed using Krippendorff alpha (αk) along with bootstrapped 95% CI. Imaging was reviewed a second time by all evaluators ~1 month later. All imaging was blinded and randomized. Intraobserver reproducibility was assessed for the primary classifications using Fleiss kappa and subclassification reproducibility was assessed by Krippendorff alpha (αk) along with 95% CI. Interpretations for reliability estimates were based on Landis and Koch (1977): 0 to 0.2, slight; 0.2 to 0.4, fair; 0.4 to 0.6, moderate; 0.6 to 0.8, substantial; and >0.8, almost perfect agreement. Twenty-five cases were read for a total of 225 initial and 225 repeated evaluations. Adjusted interobserver reliability was almost perfect (kF=0.82; CI, 0.77-0.87) across all raters. Subclassification reliability was substantial (αK=0.79; CI, 0.62-0.90). Adjusted intraobserver reproducibility was almost perfect (kF=0.81; CI, 0.71-0.90) for both primary classifications and for subclassifications (αk=0.81; CI, 0.73-0.86). The reliability for the AOSpine thoracolumbar spine injury slassification System was high amongst POSNA surgeons when applied to pediatric patients. Given a lack of a uniform classification in the pediatric population, the AOSpine thoracolumbar spine injury classification system has the potential to be used as the first universal spine fracture classification in children. Level III.

Sustained Virologic Response Rates of Sofosbuvir and Velpatasvir in Patients with Hepatitis C Genotype 3: A Meta-Analysis

Context Patients with hepatitis C virus (HCV) genotype 3 are regarded as a difficult-to-cure population in an era of direct-acting antiviral treatment.ObjectivesThis meta-analysis aimed to evaluate sustained virologic response rates resulting from a fixed-dose combination of sofosbuvir (SOF) and velpatasvir (VEL), also known as Epclusa® (Gilead, Forster City, CA) in patients with HCV genotype 3.MethodsIn this study, we searched PubMed/MEDLINE, Embase, and the Cochrane Library for relevant studies from inception until May 3rd, 2019. The primary outcome measure used was the rate of the sustained virological response at week-12 (SVR12) post-treatment. The heterogeneity of results was evaluated, and influence analyses were performed using the R software. In addition, publication bias was assessed using the funnel and Egger tests.ResultsEleven trials (n = 2246 patients) were included in this meta-analysis. The pooled SVR12 rate in patients with HCV-genotype 3 (GT3) was 94.6% with a random effect model by inverse method (95% Cl 92.5% - 96.1%, I2 = 53%, P = 0.02). A subpopulation analysis indicated that the pooled SVR12 rates were 96.3% in GT3 patients with compensated cirrhosis and 94.0% in GT3 patients with prior anti-HCV treatment. Moreover, influence analysis suggested that the most significant source of heterogeneities resulted from one trial, which enrolled most patients with HCV subtype 3b.ConclusionsOur meta-analysis showed a high SVR12 rate of SOF/VEL in GT3 patients regardless of compensated cirrhosis the status and/or a history of previous interferon-based treatments. These results highlight the need for more trials investigating the effectiveness of the SOF/VEL regimen in patients with HCV subtype 3b.

Simvastatin Improves Behavioral Disorders and Hippocampal Inflammatory Reaction by NMDA-Mediated Anti-inflammatory Function in MPTP-Treated Mice.

The cognitive function impairment may be related to the inflammation of the hippocampus in Parkinson's disease. Simvastatin can play a positive role in Parkinson's disease. The purpose of this study was to investigate whether simvastatin could improve behavioral disorders, especially depression, anxiety and cognitive function in mouse PD models, and further explore the molecular mechanism. In the present study, C57BL-6 mice underwent intraperitoneal injection of MPTP (30mg/kg) once a day for 5 consecutive days. At the same time, simvastatin (10mg/kg) was pretreated for 2days before the Parkinson's disease model was established, and then continued for 5days, and the control group underwent intraperitoneal injection of MK801 (dizocilpine, 0.2mg/kg) and saline solution. Depression status was tested by a tail suspension test and a sucrose splash test, followed by an open-field test and an elevated plus maze test to determine anxiety levels. Spatial behavior and muscle status were measured with a water maze and a rotarod test. The expression of RNA and protein of N-methyl-D-aspartate receptor subtype 2B (NMDAR2B), nerve growth factor IB (Nur77), cyclooxygenase-2 (COX-2), and tumor necrosis factor (TNF) α were assayed by real-time polymerase chain reaction and Western blot. Our results showed that simvastatin can improve the cognitive function, anxiety, and depression of PD mice with MPTP injury. Simvastatin reversed the NMDAR2B increase, restored Nur77 downward, and reduced the expression of COX-2 and TNF-α in MPTP-treated mice. This role of simvastatin was consistent with MK801 in increasing the expression of Nur77 and inhibiting NMDAR2B and cytokines in MPTP-lesioned PD mice. These findings suggest that reversed the NMDAR2B increase, restored Nur77 downward, and reduced the expression of COX-2 and TNF-α in MPTP-treated mice may be one of the mechanisms that simvastatin improves cognitive functions, depression, and anxiety in MPTP-lesioned mice.

Impact of hepatitis C virus genotype 3 on liver disease progression in a Chinese national cohort.

Hepatitis C virus (HCV) genotype 3, particularly subtype 3b, is increasing in prevalence and distribution in China. This study evaluated the prevalence, regional distribution, clinical characteristics, host factors, treatment outcomes, and disease progression of patients with HCV genotype 3 in China. A 5-year follow-up was preceded by a cross-sectional study. Treatment choices were at the discretion of treating physicians. Estimated infection time to overall-disease-progression (defined by ≥1 of: newly diagnosed cirrhosis; cirrhosis at baseline, Child-Turcotte-Pugh score increased 2 points or more; progression from compensated cirrhosis to decompensated cirrhosis; hepatocellular carcinoma; liver transplantation; or death) was calculated using the Kaplan-Meier method. Cox regression analyses were conducted to evaluate the risk factors for disease progression. The cross-sectional study enrolled 997 patients, including 91 with HCV genotype 3 infection. Among them, subtype 3b (57.1%) was more dominant than subtype 3a (38.5%). Five hundred and twelve patients were included into the follow-up phase. Among patients analyzed for estimated infection time to overall-disease-progression, 52/304 (17.1%) patients with HCV genotype 1 and 4/41 (9.8%) with HCV genotype 3 (4/26 with genotype 3b, 0/13 with genotype 3a, and 0/2 with undefined subtype of genotype 3) experienced overall-disease-progression. Patients with HCV genotype 3 were younger than those with genotype 1 (mean age: 39.5 ± 8.7 vs. 46.9 ± 13.6 years) and demonstrated more rapid disease progression (mean estimated infection time to overall-disease-progression 27.1 vs. 35.6 years). HCV genotype 3, specifically subtype 3b, is associated with more rapid progression of liver disease. Further analysis to compare HCV subtype 3a and 3b is needed in high prevalence regions. NCT01293279, https://clinicaltrials.gov/ct2/show/NCT01293279; NCT01594554, https://clinicaltrials.gov/ct2/show/NCT01594554.

Remifentanil protects neurological function of rats with cerebral ischemia-reperfusion injury via NR2B/CaMKIIα signaling pathway.

The effects of remifentanil were studied on cerebral ischemia-reperfusion injury (IRI) in rats, and its regulatory effect was explored on the N-methyl D-aspartate receptor subtype 2B (NR2B)/calcium/calmodulin-dependent protein kinase type II subunit alpha (CaMKII) signaling pathway in cerebral tissues. A total of 40 Sprague-Dawley rats were randomly assigned into sham group, model group, low-dose group (remifentanil injected into the caudal veins at 2 μg/kg) and high-dose group (remifentanil injected into the caudal veins at 10 μg/kg). Then, in the model, low-dose and high-dose groups, the rat model of cerebral IRI was established through middle cerebral artery occlusion (namely, ischemia for 1 h and reperfusion for 2 h), while no thread was inserted into the rats in the sham group. Neurological function of rats in each group was evaluated, and the cerebral infarct size was measured using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Moreover, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was performed to determine the neuronal apoptosis level, and mitochondrial membrane potential was measured via JC-1 assay. Moreover, the reactive oxygen species (ROS) level in neurons and the malondialdehyde (MDA) content in cerebral tissues were determined using 2',7'-dichlorofluorescin diacetate (DCFH-DA) assay and enzyme-linked immunosorbent assay (ELISA), respectively. Finally, the expression levels of apoptosis-associated proteins and the NR2B/CaMKIIα signaling pathway-associated proteins in cerebral tissues were measured by Western blotting. Remifentanil preconditioning substantially decreased the neurological score of rats (p<0.01), cerebral infarct size (p<0.01), neuronal apoptosis level (p<0.01), ROS level in neurons (p<0.01), MDA content (p<0.01) and expression levels of cysteinyl aspartate specific proteinase-3 (Caspase-3), NR2B, phosphorylated CaMKIIα (p-CaMKIIα) and p-cAMP responsive element binding protein (p-CREB) (p<0.01), but it increased the mitochondrial membrane potential (p<0.01) and B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (Bax) ratio (p<0.01). Remifentanil can repress the NR2B/ CaMKIIα signaling pathway in the neurons of rats with cerebral IRI to decrease the p-CREB expression, ROS level and MDA content in neurons, neuronal apoptosis level and cerebral infarct size, and increase the mitochondrial membrane potential, thereby protecting the neurological function.

Retreatment of patients with chronic hepatitis C, subtype 1b and cirrhosis, who failed previous direct-acting antiviral therapy including first- and second-generation NS5A inhibitors with ombitasvir/paritaprevir/ritonavir, dasabuvir+sofosbuvir+ribavirin.

The use of direct-acting antiviral agents (DAAs) in patients with chronic HCV genotype 1 infection results in sustained virologic response (SVR) rates of 95%-97%, but 3%-5% of patients experience virologic failure. We observed 17 patients infected with HCV subtype 1b who failed previous treatment with DAA, including 13 subjects (76.5%) with liver cirrhosis. Twelve subjects (70.6%) previously received NS5A inhibitors of the first generation (ledipasvir or daclatasvir) and five subjects (29.4%) - the second generation (velpatasvir). All patients were retreated with a combination of ombitasvir/paritaprevir/ritonavir and dasabuvir (3D) with sofosbuvir (SOF) and ribavirin (RBV). We compared SVR12 rates depending on fibrosis stage, presence of just single or double NS5A mutations (L31M/V/I and/or Y93H), and on the generation of previously used NS5A inhibitor. Observed SVR12 rates were as follows: 94.1% (16/17 patients) overall; 100% in patients without cirrhosis (n=4) vs 92.3% in those with cirrhosis (n=13); 100% with single L31M/V/I or Y93H mutation (n=7) vs 88.9% with double mutations (n=9); 100% in patients who previously failed first generation (n=12) vs 80.0% in those failed second-generation NS5A inhibitors (n=5). Retreatment with 3D+0SOF+RBV was highly effective and safe in patients with chronic HCV GT1b infection who failed previous use of NS5A inhibitors. Fibrosis stage, baseline presence of NS5A RAS mutations and the generation of previously used NS5A inhibitors may impact the probability of achieving SVR12 , but statistical significance was not demonstrated in our small retrospective cohort. Further studies in a larger population are needed to confirm or not the predictive value of these baseline factors.