ABSTRACT
ABSTRACT: Levosimendan, a calcium sensitizer, exerts inotropic action through improving left ventricular ejection fraction. We noticed that only few clinical studies are published in which the effects of levosimendan on cardiac function are studied by echocardiography. When screening the literature (PubMed, Embase, and CENTRAL, from inception to August 2020), we found 29 randomized controlled trials on levosimendan containing echocardiographic data. We included those studies, describing a total of 574 heart failure patients, in our meta-analysis and extracted 14 ultrasonic parameters, pooling the effect estimates using a random-effect model. Our analysis of the diastolic parameters of the left ventricle shows that levosimendan reduce the early/late transmitral diastolic peak flow velocity ratio [standardized mean difference (SMD) -0.45 to 95% confidence interval (CI) (-0.87 to -0.03), P = 0.037] and E/e' (e': mitral annulus peak early diastolic wave velocity using tissue-doppler imaging) [SMD -0.59, 95% CI (-0.8 to -0.39), P < 0.001]. As it regards the systolic parameters of the right ventricle, levosimendan increased tricuspid annular plane systolic excursion [SMD 0.62, 95% CI (0.28 to 0.95), P < 0.001] and tricuspid annular peak systolic velocity [SMD 0.75, 95% CI (0.35 to 1.16), P < 0.001], and reduced systolic pulmonary artery pressure [SMD -1.02, 95% CI (-1.32, -0.73), P < 0.001]. As it regards the diastolic parameters of the right ventricle, levosimendan was associated with the decrease of Aa (peak late diastolic tricuspid annular velocity using tissue-doppler imaging) [SMD -0.38, 95% CI (-0.76 to 0), P = 0.047] and increase of Ea (peak early diastolic tricuspid annular velocity using tissue-doppler imaging) [SMD 1.03, 95% CI (0.63 to 1.42), P < 0.001] and Ea/Aa [SMD 0.86, 95% CI (0.18 to 1.54), P = 0.013]. We show that levosimendan is associated with an amelioration in the diastolic and systolic functions of both ventricles in heart failure patients.
Subject(s)
Cardiotonic Agents/pharmacology , Heart Failure/drug therapy , Simendan/pharmacology , Diastole/drug effects , Echocardiography , Heart Failure/physiopathology , Humans , Randomized Controlled Trials as Topic , Stroke Volume/drug effects , Systole/drug effects , Ventricular Function, Left/drug effects , Ventricular Function, Right/drug effectsABSTRACT
UNLABELLED: Background and aims. CD4+ T cells play an important role in response to hepatitis B virus (HBV) infection. We investigated the change in CD4+ T-cell subpopulations and viral load in patients with chronic HBV infection who were treated with entecavir. MATERIAL AND METHODS: Thirty patients with chronic HBV infection were enrolled according to the criteria recommended by the Chinese Society of Infectious Diseases and the Chinese Society of Hepatology. The expressions of signature transcription factors and cytokines of CD4+ T-cell subpopulations were measured in chronic hepatitis B (CHB) patients treated with entecavir treatment. RESULTS: Entecavir treatment significantly attenuated hepatitis B virus DNA load and affected the CD4+ T-cell subsets in CHB patients. A dramatic decrease in the Th17 and Treg cell frequencies and expressions of their related cytokines were found in CHB patients with entecavir treatment. In contrast, entecavir treatment caused a remarkable increase in the Th2 cell frequencies and expressions of their related cytokines. CONCLUSION: Our results suggested that Th17 and Treg cells were the more sensitive subtypes to entecavir- induced inhibition of HBV replication compared to Th1 and Th2 cells in chronic HBV patients.
Subject(s)
Antiviral Agents/therapeutic use , CD4-Positive T-Lymphocytes/immunology , Cytokines/immunology , Guanine/analogs & derivatives , Hepatitis B, Chronic/drug therapy , Adult , Case-Control Studies , DNA, Viral/blood , Female , Guanine/therapeutic use , Hepatitis B virus/genetics , Hepatitis B, Chronic/immunology , Humans , Male , Middle Aged , T-Lymphocyte Subsets/immunology , T-Lymphocytes, Regulatory/immunology , Th1 Cells/immunology , Th17 Cells/immunology , Th2 Cells/immunology , Viral LoadABSTRACT
Understanding of codon usage bias of Fritillaria cirrhosa can provide theoretical basis for heterologous biosynthesis of F. cirrhosa alkaloids by genetic engineering technology. A total of 9 843 full length coding sequences (CDS) from the F. cirrhosa transcriptome data were used for the analysis of codon usage bias. The GC and GC3s contents, effective number of codons(ENC) and relative synonymous codon usage (RSCU) were calculated using the CodonW software. The results show that the codon usage bias value is low in the CDS of F. cirrhosa. A total of 15 codons, including UUG, CUU, AUU, GUU, UCA, CCU, CCA, ACU, ACA, GCA, UAU, CAU, AAU, AGA and GGA, were identified as optimal codons in F. cirrhosa. The optimal codons generally end with A/T at the third codon position. By the transcriptome annotation, we found 26 CDSs possibly involved in the biosynthesis of alkaloids in the F. cirrhosa. The proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae are low in these CDSs. We also proposed a method for the codonoptimization in these target genes. Our work lays the foundation for further study on the biosynthesis of alkaloids of the F. cirrhosa in heterologous species.
Subject(s)
Codon , Fritillaria/genetics , Transcriptome , Alkaloids/biosynthesis , Fritillaria/chemistryABSTRACT
There are many valuable medicinal plants in Ginseng genus belonging to Araliaceae. Among them, Panax ginseng, P. quinquefolium and P. notoginseng are the most famous species. With the development of next-generation sequencing (NGS) technologies, sequencing and analysis of transcriptomes have become powerful tools for discovery of novel genes, screening molecular markers and elucidation of specific biosynthetic pathway of secondary metabolites. Their transcriptomes provided abundant genes for further study on functional genomics. Here this paper summarized the recent advances in the transcriptomic studies of these three medicinal plants, including discovery of novel genes and elucidation of metabolic regulation, which will contribute to functional genomics in ginseng species.
Subject(s)
Biosynthetic Pathways , Panax/genetics , Transcriptome , High-Throughput Nucleotide Sequencing , Panax/metabolism , Plants, Medicinal/genetics , Plants, Medicinal/metabolismABSTRACT
Natural products with complex and diverse structures are the major sources of new drugs. The biosynthesis of natural products is considered to be one of the best ways to solve the problems of complex and scarce natural products. DNA assembly technology and genome editing technology are two key technologies in the emerging interdisciplinary field of synthetic biology. A number of novel DNA assembly methods developed in the last few years have paved the way for the engineering of high molecular weight DNA molecules, including whole genomes, hence, it can realize the reconstruction of the metabolic pathways and speed up optimization process. A wide variety of new tools for microbial genome editing will be applied widely to modify the chassis genome to increase its adaptation with the exogenetic pathways. This article summarized the latest advance with respect to DNA assembly and genome editing, which aims to provide help for reconstruction and optimization of the synthetic biological systems of natural products.
Subject(s)
Biological Products , Biotechnology , Gene Editing , Synthetic Biology , DNA , Metabolic Networks and PathwaysABSTRACT
Elucidation of the biosynthetic pathways of natural products is not only the major goal of herb genomics, but also the solid foundation of synthetic biology of natural products. Here, this paper reviewed recent advance in this field and put forward strategies to elucidate the biosynthetic pathway of natural products. Firstly, a proposed biosynthetic pathway should be set up based on well-known knowledge about chemical reactions and information on the identified compounds, as well as studies with isotope tracer. Secondly, candidate genes possibly involved in the biosynthetic pathway were screened out by co-expression analysis and/or gene cluster mining. Lastly, all the candidate genes were heterologously expressed in the host and then the enzyme involved in the biosynthetic pathway was characterized by activity assay. Sometimes, the function of the enzyme in the original plant could be further studied by RNAi or VIGS technology. Understanding the biosynthetic pathways of natural products will contribute to supply of new leading compounds by synthetic biology and provide "functional marker" for herbal molecular breeding, thus but boosting the development of traditional Chinese medicine agriculture.
Subject(s)
Biological Products , Biosynthetic Pathways , Synthetic Biology , Genomics , Medicine, Chinese Traditional , Multigene Family , Plants, Medicinal/genetics , Plants, Medicinal/metabolismABSTRACT
Flavonoids are the valuable components in medicinal plants, which possess a variety of pharmacological activities, including anti-tumor, antioxidant and anti-inflammatory activities. There is an unambiguous understanding about flavonoids biosynthetic pathway, that is,2S-flavanones including naringenin and pinocembrin are the skeleton of other flavonoids and they can transform to other flavonoids through branched metabolic pathway. Elucidation of the flavonoids biosynthetic pathway lays a solid foundation for their synthetic biology. A few flavonoids have been produced in Escherichia coli or yeast with synthetic biological technologies, such as naringenin, pinocembrin and fisetin. Synthetic biology will provide a new way to get valuable flavonoids and promote the research and development of flavonoid drugs and health products, making flavonoids play more important roles in human diet and health.
Subject(s)
Biosynthetic Pathways , Flavanones/biosynthesis , Flavonoids/biosynthesis , Synthetic Biology , HumansABSTRACT
Taxol, a kind of terpenoid secondary metabolite produced by Taxus brevifolia, is an effective anticancer drug that manufacture relies mainly on the extraction form plants. In order to solve the resource shortage, a lot of work has been done to develop the alternative method. Recently, using synthetic biology to realize heterologous biosynthesis of the precursors of taxol has become a hotspot. Now, the basic framework of taxol biosynthetic pathways has been confirmed, and most enzyme genes involved in taxol biosynthesis have been cloned and identified. The two taxol precursors, taxa-4(5),11(12)-diene and taxa-4(20),11(12)-dien-5α-ol, have been synthesized in Escherichia coli and Saccharomyces cerevisiae. Here this paper reviewed the recent advances in the biosynthetic pathway of taxol and the latest developments of synthetic biology, which aims to provide a guidance for the heterologous biosynthesis of taxol.
Subject(s)
Biosynthetic Pathways , Paclitaxel/biosynthesis , Synthetic Biology , Taxus/chemistry , TerpenesABSTRACT
This study aimed to provide guidance for the heterogenous gene expression, gene prediction and species evolution by analyzing codon usage bias of Catharanthus roseus.The codon composition and usage bias of 30 437 high-confidence coding sequences from C.roseus were analyzed and the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 25 genes involved in the biosynthesis of terpenoid indole alkaloids (TIAs) in C.roseus were calculated.The results showed that the average GC content of the genes was 42.47%; the average GC content of the third bases in codon was 35.89%.The relative synonymous codon usage (RSCU) of 28 codons were greater than 1 and 26 of them ended with A or T.The above 25 genes involved in TIA biosynthesis contained much more rare condons of E.coli than that of S.cerevisiae.It was concluded that C.roseus mainly prefered the codons ending with A or T and the rule of codon usage was more different to E.coli than S.cerevisiae.Thus, S.cerevisiae may be more suitable host for heterologous expression of these genes.
Subject(s)
Catharanthus/genetics , Codon , Base Composition , Escherichia coli/genetics , Plants, Medicinal/genetics , Saccharomyces cerevisiae/geneticsABSTRACT
Catharanthus roseus can produce a variety of terpenoid indole alkaloids (TIA), most of which exhibit strong pharmacological activities. Hence, biosynthesis and regulation of TIA have received recent attention. 3α (S)-strictosidine is an important node in TIA biosynthesis, which is a condensation product of secologanin and tryptamine. The former is produced in iridoid pathway, and the latter is produced in indole pathway. Vindoline and catharanthine, which are produced respectively by 3α (S)-strictosidine via multi-step enzymatic reaction, can form α-3, 4-anhydrovinblastine by the condensation reaction. Then, vinblastine and vincristine are generated from α-3, 4-anhydrovinblastine. Many transcription factors are involved in the regulation of TIA synthesis, such as AP2/ERF and WRKY. Illumination of biosynthetic pathway has laid a foundation for the study of synthetic biology. Today, 3α (S)-strictosidine and vindoline have been synthesized in heterologous hosts Saccharomyces cerevisiae.Research about synthetic biology and the regulation mechanisms will provide a guidance for the production and development of TIA drugs in C. roseus.
Subject(s)
Biosynthetic Pathways , Catharanthus/metabolism , Secologanin Tryptamine Alkaloids/metabolism , Transcription Factors/metabolismABSTRACT
BACKGROUND: Asymptomatic hyperuricemia (HUA) and normouricemic gout are common in clinic but recommendations for them in hypertension management are absent. The present study aims to simultaneously evaluate the effect of HUA and gout on long-term mortality in hypertension. METHODS: Individuals from 2007-2018 National Health and Nutrition Examination Survey were enrolled. Hazard ratios and 95% confidence intervals (CIs) were calculated with the aid of the Cox proportional-hazards model. The restricted cubic spline (RCS) analysis was made to show the dose-response relationship between uric acid and mortality. All-cause mortality and cardiovascular mortality were compared using the Kaplan-Meier curve with a log-rank test. RESULTS: Thirty thousand eight hundred and nineteen eligible individuals were included, of which 5841 suffered from HUA and 1476 suffered from gout. During a median follow-up of 7.25 (95% CI 7.18-7.32) years, 2924 (6.8%) patients died, including 722 (1.6%) cases of cardiovascular death. Hypertensive patients with HUA and gout showed 1.34 and 1.29 times higher all-cause mortality compared with those without HUA or gout. For hypertensive patients without gout, HUA was significantly associated with higher risk of all-cause [1.27 (1.13, 1.43)] and cardiovascular [1.80 (1.44, 2.24)] mortality compared with normouricemia. However, for hypertensive patients without HUA, gout was associated with a higher mortality but not statistically significant. A J-shaped relationship was found between serum uric acid and mortality. CONCLUSION: HUA and gout are additive risk factors for all-cause and cardiovascular mortality in hypertension. Furthermore, asymptomatic HUA is significantly associated with poor long-term prognosis but normouricemic gout is not.
Subject(s)
Gout , Hypertension , Hyperuricemia , Nutrition Surveys , Humans , Gout/mortality , Gout/complications , Gout/epidemiology , Hyperuricemia/complications , Hyperuricemia/epidemiology , Hyperuricemia/mortality , Male , Female , Middle Aged , Hypertension/mortality , Hypertension/epidemiology , Hypertension/complications , Adult , Risk Factors , Aged , Uric Acid/bloodABSTRACT
CONTEXT: While the association between n-3 polyunsaturated fatty acids (PUFAs) and cardiovascular (CV) events has been thoroughly examined, there is still a scarcity of research regarding their effect on the long-term prognosis in diabetic patients. OBJECTIVE: We aimed to explore the effects of n-3 PUFA on all-cause and cardiovascular mortality in patients with pre-diabetes and diabetes. METHOD: Herein, a total of 16 539 eligible individuals were enrolled from the National Health and Nutrition Examination Survey (NHANES) 2003 to 2018, and categorized into T1, T2, and T3 based on the tertiles of n-3 PUFA. The Cox proportional risk regression models, Kaplan-Meier curve, and subgroup analysis were conducted to evaluate the association between n-3 PUFA and mortality. Restricted cubic spline (RCS) curves graphically demonstrated the dose-response relationship. Additionally, weighted quantile sum (WQS) models were adopted to measure the mixed and individual effects of n-3 PUFA on mortality. RESULTS: Following a median follow-up period of 8.42 years, 3010 individuals died, with 989 deaths attributed to CV diseases. Significantly lower risk of all-cause (T2: 0.81 [0.71-0.92], T3: 0.77 [0.64-0.94]) and CV (T2: 0.75 [0.61-0.93]) mortality was observed after adjusting for multivariables compared to the reference (T1). Meanwhile, the RCS curve revealed a negative nonlinear association between n-3 PUFA and mortality. None of the interactions in any subgroup analysis were statistically significant except for BMI (P for interaction = .049). Finally, the WQS analysis demonstrated alpha-linolenic acid (ALA) and docosapentaenoic acid (DPA) as the main contributors to n-3 PUFAs' benefits against mortality. CONCLUSION: Increased dietary intake of n-3 PUFAs, particularly ALA and DPA, was associated with a reduced risk of all-cause and CV mortality among Americans with prediabetes and diabetes.
Subject(s)
Cardiovascular Diseases , Fatty Acids, Omega-3 , Nutrition Surveys , Prediabetic State , Humans , Prediabetic State/mortality , Prediabetic State/epidemiology , Prediabetic State/complications , Female , Male , Fatty Acids, Omega-3/administration & dosage , Cardiovascular Diseases/mortality , Cardiovascular Diseases/prevention & control , Middle Aged , Adult , Aged , Follow-Up Studies , Diabetes Mellitus/mortality , Diabetes Mellitus/epidemiology , Cause of Death , Prognosis , DietABSTRACT
Background: There are various cross-sectional studies that concluded that vitamin D is associated with blood pressure, but randomized controlled studies have not yielded consistent conclusions. Considering many limitations indeed, our study aimed to examine whether concentrations of 25(OH)D are inversely associated with blood pressure in people without a previous diagnosis of hypertension. Method: We analyzed data from the 2005-2018 National Health and Nutrition Examination Survey. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by applying multivariable logistic regression models. The dose-response relationship was assessed by means of restricted cubic spline regression, and stratification analyses were employed to test the consistency between the subgroups. Results: Of 17,467 participants aged ≥ 20 years without a previous diagnosis of hypertension, 4,769 had higher blood pressure. Compared with individuals whose 25(OH)D levels were in the bottom quartile (<44.3 nnol/L), adjusting for multiple confounders, the ORs for higher blood pressure were 0.90(95%CI 0.78, 1.05), 0.85(95%CI 0.72, 0.99), and 0.86(95%CI 0.72, 1.02), respectively (P for trend = 0.096). Furthermore, as a continuous variable, 25(OH)D concentrations were non-linearly associated with an increased risk of hypertension (P < 0.001). The interaction between the sleeplessness subgroup and higher blood pressure was significant (P = 0.042). Conclusion: In adults without a previous diagnosis of hypertension in the United States, concentrations of 25(OH)D were inversely associated with higher blood pressure when it was <84 nmol/L.
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OBJECTIVE: Levosimendan, an inotrope, is widely used in the management of heart failure (HF) and cardiac surgery, but it remains uncertain whether levosimendan can improve renal function in patients with left ventricular dysfunction (LVD). METHODS: PubMed, Embase, and Cochrane CENTRAL from the inception to June 2020 were systematically screened for randomized controlled trials (RCTs) to investigate whether levosimendan offers kidney-related advantages in cardiovascular patients with LVD. We pooled the effects using a random-effect model. RESULTS: Twenty-eight studies enrolling 5069 patients were included. Levosimendan reduced the sCr (SMD -0.28, 95% CI (-0.48, -0.09), P = 0.005, I2 = 52.5%, high quality) and the risk of ARF (relative risk 0.75, 95%CI (0.60, 0.95), P = 0.017, I2 = 11.3%, moderate-quality) in patients with LVD compared with control group. The reduction of sCr was more pronounced in patients with a relatively higher baseline sCr level. For secondary outcomes, levosimendan therapy was associated with the improvement of GFR (SMD 0.32, 95%CI (-0.05, 0.68), P = 0.092, I2 = 55.1%, low-quality) and urine output (SMD 0.42, 95%CI (0.06, 0.79), P = 0.024, I2 = 50.0%, very low-quality), but there was no significant reduction in BUN (SMD -0.14, 95%CI (-0.97, 0.70), P = 0.774, I2 = 77.9%, very low-quality). CONCLUSIONS: Levosimendan might improve renal function of patients with LVD.
Subject(s)
Cardiotonic Agents/administration & dosage , Simendan/administration & dosage , Ventricular Dysfunction, Left/drug therapy , Cardiotonic Agents/pharmacology , Glomerular Filtration Rate , Humans , Kidney/drug effects , Kidney/metabolism , Kidney Function Tests , Randomized Controlled Trials as Topic , Simendan/pharmacology , Ventricular Dysfunction, Left/physiopathologyABSTRACT
BACKGROUND: Major histocompatibility complex class I-related chain A (MICA) is considered as a tumor antigen, and its expression is affected by its genetic polymorphisms. However, the relationship between rs2596542 polymorphisms in MICA promoter region and hepatocellular carcinoma (HCC) is not fully elucidated so far. This study aims to explore the relationship between single nucleotide polymorphism of rs2596542 and the risk of HCC development through meta-analysis. METHODS: MEDLINE, Web of Science, and EMBASE databases were systematically searched to identify relevant studies. A meta-analysis was performed to examine the association between MICA rs2596542 polymorphism and susceptibility to HCC. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. RESULTS: Fourteen case-control studies involving 4,900 HCC cases and 19,519 controls were included. The MICA rs2596542C allele was significantly associated with decreased risk of HCC based on allelic contrast (ORâ=â0.76, 95% CIâ=â0.69-0.83, Pâ<â.001), homozygote comparison (ORâ=â0.57, 95% CIâ=â0.48-0.69, Pâ<â.001), and a recessive genetic model (ORâ=â0.77, 95% CIâ=â0.65-0.91, Pâ<â.001), whereas patients carrying the MICA rs2596542TT genotype had significantly higher risk of HCC than those with the CT or CC genotype (TT vs CTâ+âCC, ORâ=â1.57, 95% CIâ=â1.36-1.81, Pâ<â.001). Subgroups analyses based on the ethnic or the source of control groups found very similar findings. CONCLUSION: The C allele in MICA rs2596542 is a protective factor for hepatocarcinogenesis, whereas the T allele is a risk factor. Further large and well-designed studies are needed to confirm this conclusion.
Subject(s)
Carcinoma, Hepatocellular/genetics , Histocompatibility Antigens Class I/genetics , Liver Neoplasms/genetics , Humans , Polymorphism, Single NucleotideABSTRACT
BACKGROUND: Proton pump inhibitors (PPIs) are usually prescribed to protect against gastrointestinal bleeding in patients on dual antiplatelet therapy. This meta-analysis reviewed clinical outcomes in patients taking aspirin and clopidogrel, with and without concomitant PPIs to address concerns of adverse reactions. METHODS: We searched PubMed, Embase, and the Cochrane Library for articles published between January 1, 2010 and April 11, 2017. The primary end points were major adverse cardiovascular events and gastrointestinal bleeding. Secondary end points were myocardial infarction, stent thrombosis, revascularization, cardiogenic death, and all-cause mortality. RESULTS: The meta-analysis included 33,492 patients in 4 randomized controlled trials and 8 controlled observational studies. Overall, patients taking PPIs had statistical differences in major adverse cardiovascular events [odds ratio (OR) 1.17 (95% confidence interval [CI] 1.07-1.28); Pâ=â.001; Iâ=â28.3%], gastrointestinal bleeding [OR 0.58 (95% CI 0.36-0.92); Pâ=â.022; Iâ=â80.6%], stent thrombosis [OR 1.30 (95% CI 1.01-1.68); Pâ=â.041; Iâ=â0%], and revascularization [OR 1.20 (95% CI 1.04-1.38); Pâ=â.011; Iâ=â5.1%], compared those not taking PPIs. There were no significant differences in myocardial infarction [OR 1.03 (95% CI 0.87-1.22); Pâ=â.742; Iâ=â0%], cardiogenic death [OR 1.09 (95% CI 0.83-1.43); Pâ=â.526; Iâ=â0%], or all-cause mortality [OR 1.08 (95% CI 0.93-1.25); Pâ=â.329; Iâ=â0%). CONCLUSIONS: Among the patients taking aspirin and clopidogrel, the results indicated that the combined use of PPIs increased the rates of major adverse cardiovascular events, stent thrombosis, and revascularization.
Subject(s)
Aspirin/therapeutic use , Coronary Disease/drug therapy , Platelet Aggregation Inhibitors/therapeutic use , Proton Pump Inhibitors/adverse effects , Ticlopidine/analogs & derivatives , Clopidogrel , Humans , Ticlopidine/therapeutic useABSTRACT
Epigenetics may affect the susceptibility for type 2 diabetes mellitus (T2DM). Aberrant DNA methylation patterns are nowadays recognized as a key epigenetic hallmark of T2DM. Previously, our studies have shown that the hypomethylation of human miR-375 promoter may contribute to the pathogenesis of T2DM. However, no comprehensive study defines the miR-375 promoter methylation patterns present in the established pancreatic ß cell line. To address this matter, we have analyzed the DNA methylation profile of insulinoma MIN6 cells by MassARRAY spectrometry and employed the DNA demethylating drug 5-aza-2'-deoxycytidine (5-aza-CdR) to treat MIN6 cells to explore the methylation patterns of the mmu-miR-375. The expression of mmu-miR-375 in mRNA level was measured by quantitative RT-PCR (qRT-PCR). Methylation analysis reveals that MIN6 cells display hypermethylation at the mmu-miR-375 promoter. Following the decreased methylation of mmu-miR-375, the relative expression of mmu-miR-375 increased gradually after 5-Aza-CdR treatment. In addition, we find that there was an inverse correlation between DNA methylation levels and transcription level of mmu-miR-375. In summary, this is the first report for analyzing mmu-miR-375 promoter methylation using MALDI-TOF MS technology and our results indicate that promoter hypermethylation of the mmu-miR-375 is a common event in MIN6 cells.
ABSTRACT
AIM: To investigate the mechanism by which hepatitis C virus (HCV) core protein-induced miR-93-5p up-regulation regulates the interferon (IFN) signaling pathway. METHODS: HCV-1b core protein was exogenously expressed in Huh7 cells using pcDNA3.1 (+) vector. The expression of miR-93-5p and interferon receptor 1 (IFNAR1) was measured using quantitative reverse transcription-polymerase chain reaction and Western blot. The protein expression and phosphorylation level of STAT1 were evaluated by Western blot. The overexpression and silencing of miR-93-5p and IFNAR1 were performed using miR-93-5p agomir and antagomir, and pcDNA3.1-IFNAR1 and IFNAR1 siRNA, respectively. Luciferase assay was used to identify whether IFNAR1 is a target of miR-93-5p. Cellular experiments were also conducted. RESULTS: Serum miR-93-5p level was increased in patients with HCV-1b infection and decreased to normal level after HCV-1b clearance, but persistently increased in those with pegylated interferon-α resistance, compared with healthy subjects. Serum miR-93-5p expression had an AUC value of 0.8359 in distinguishing patients with pegylated interferon-α resistance from those with pegylated interferon-α sensitivity. HCV-1b core protein increased miR-93-5p expression and induced inactivation of the IFN signaling pathway in Huh7 cells. Furthermore, IFNAR1 was identified as a direct target of miR-93-5p, and IFNAR1 restore could rescue miR-93-5p-reduced STAT1 phosphorylation, suggesting that the miR-93-5p-IFNAR1 axis regulates the IFN signaling pathway. CONCLUSION: HCV-1b core protein-induced miR-93-5p up-regulation inhibits the IFN signaling pathway by directly targeting IFNAR1, and the miR-93-5p-IFNAR1 axis regulates STAT1 phosphorylation. This axis may be a potential therapeutic target for HCV-1b infection.
Subject(s)
Hepacivirus/metabolism , Hepatitis C/metabolism , Hepatocytes/metabolism , MicroRNAs/metabolism , Receptor, Interferon alpha-beta/metabolism , Signal Transduction , Viral Core Proteins/metabolism , Adult , Antiviral Agents/therapeutic use , Cell Line, Tumor , Drug Resistance, Viral , Female , HEK293 Cells , Hepacivirus/drug effects , Hepacivirus/pathogenicity , Hepatitis C/blood , Hepatitis C/drug therapy , Hepatitis C/virology , Hepatocytes/drug effects , Hepatocytes/virology , Host-Pathogen Interactions , Humans , Interferon-alpha/therapeutic use , Male , MicroRNAs/genetics , Middle Aged , Phosphorylation , Receptor, Interferon alpha-beta/drug effects , Receptor, Interferon alpha-beta/genetics , STAT1 Transcription Factor/metabolism , Signal Transduction/drug effects , Up-Regulation , Viral Core Proteins/geneticsABSTRACT
OBJECTIVE: To study genotype distribution and the characteristics of hepatitis B virus (HBV) in Uighur patients with chronic hepatitis B (CHB) in Xinjiang, China. METHODS: Type specific primers and PCR were used to detect the HBV genotypes of 127 Uighur CHB patients in Xinjiang. Genotyping results were confirmed by PCR product sequencing. RESULTS: Among the 127 patients, the proportions of genotype D, B, C and B/D, C/D, B/C/D were 39.4% (50/127), 22.0% (28/127), 16.5% (21/127) and 9.4% (12/127), 8.7% (11/127) and 3.9% (5/127), respectively. The distribution of the HBV genotypes showed no significant differences between male and female patients (x2 = 8.058, P > 0.05), between HBeAg positive and negative patients (x2 = 6.033, P > 0.05), and between patients of different ages (x2 = 3.137, P > 0.05). CONCLUSION: Genotype D HBV is predominant in Uighur patients with chronic hepatitis B in Xinjiang. The distribution of various HBV genotypes shows no significant differences between these Uighur patients with different HBeAg positivity, sex and age.
Subject(s)
Genotype , Hepatitis B virus/genetics , Hepatitis B, Chronic/virology , Adolescent , Adult , Child , Child, Preschool , China/epidemiology , DNA, Viral , Female , Genome, Viral , Hepatitis B, Chronic/epidemiology , Humans , Male , Middle Aged , Young AdultABSTRACT
We report the complete chloroplast genome sequence of Gynostemma pentaphyllum, a well-known traditional Chinese medicine, which produces triterpenoid saponins similar to Panax ginseng. The assembled chloroplast genome (cpDNA) was 157,654 bp in length and structurally divided into four distinct regions, namely, large single copy region (86,794 bp), small single copy region (18,654 bp) and a pair of inverted repeat regions (26,103 bp). A total of 143 genes were annotated, including 87 protein-coding genes, 10 tRNA genes and 46 rRNA genes. Phylogenetic analysis revealed that the chloroplast genome sequence of G. pentaphyllum is most closely related to Cucumis melo.