Search

Coronaviruses (CoVs) comprise a group of important human and animal pathogens. Despite extensive research in the past 3 years, the host innate immune defense mechanisms against CoVs remain incompletely understood, limiting the development of effective antivirals and non-antibody-based therapeutics. Here, we performed an integrated transcriptomic analysis of porcine jejunal epithelial cells infected with porcine epidemic diarrhea virus (PEDV) and identified cytidine/uridine monophosphate kinase 2 (CMPK2) as a potential host restriction factor. CMPK2 exhibited modest antiviral activity against PEDV infection in multiple cell types. CMPK2 transcription was regulated by interferon-dependent and interferon regulatory factor 1 (IRF1)-dependent pathways post-PEDV infection. We demonstrated that 3'-deoxy-3',4'-didehydro-cytidine triphosphate (ddhCTP) catalysis by Viperin, another interferon-stimulated protein, was essential for CMPK2's antiviral activity. Both the classical catalytic domain and the newly identified antiviral key domain of CMPK2 played crucial roles in this process. Together, CMPK2, viperin, and ddhCTP suppressed the replication of several other CoVs of different genera through inhibition of the RNA-dependent RNA polymerase activities. Our results revealed a previously unknown function of CMPK2 as a restriction factor for CoVs, implying that CMPK2 might be an alternative target of interfering with the viral polymerase activity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Zhu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In search of more efficacious antitumor agents, a series of novel dehydroabietinol derivatives containing a triazole moiety was synthesized, and evaluated for cytotoxicity against four human cancer cell lines. Many exhibited superior cytotoxic profiles compared to the parent molecule, dehydroabietic acid. In particular, compounds 5g, 5i and 5j exhibited promising cytotoxicity with IC 50 values ranging from 4.84 to 9.62 μM against all the test cell lines. Cell clone formation and migration tests of compound 5g showed that it not only effectively inhibited the formation of MGC-803 cell colonies but also inhibited the MGC-803 cell migration and invasion. Additionally, the preliminary pharmacological mechanism indicated compound 5g induced apoptosis, arrested the mitotic process at the G0/G1 phase of the cell cycle, reduced the mitochondrial membrane potential, and increased the intracellular ROS and Ca 2+ levels., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Swine coronavirus-porcine epidemic diarrhea virus (PEDV) with specific susceptibility to pigs has existed for decades, and recurrent epidemics caused by mutant strains have swept the world again since 2010. In this study, single-cell RNA sequencing was used to perform for the first time, to our knowledge, a systematic analysis of pig jejunum infected with PEDV. Pig intestinal cell types were identified by representative markers and identified a new tuft cell marker, DNAH11. Excepting enterocyte cells, the goblet and tuft cells confirmed susceptibility to PEDV. Enrichment analyses showed that PEDV infection resulted in upregulation of cell apoptosis, junctions, and the MAPK signaling pathway and downregulation of oxidative phosphorylation in intestinal epithelial cell types. The T cell differentiation and IgA production were decreased in T and B cells, respectively. Cytokine gene analyses revealed that PEDV infection downregulated CXCL8, CXCL16, and IL34 in tuft cells and upregulated IL22 in Th17 cells. Further studies found that infection of goblet cells with PEDV decreased the expression of MUC2, as well as other mucin components. Moreover, the antimicrobial peptide REG3G was obviously upregulated through the IL33-STAT3 signaling pathway in enterocyte cells in the PEDV-infected group, and REG3G inhibited the PEDV replication. Finally, enterocyte cells expressed almost all coronavirus entry factors, and PEDV infection caused significant upregulation of the coronavirus receptor ACE2 in enterocyte cells. In summary, this study systematically investigated the responses of different cell types in the jejunum of piglets after PEDV infection, which deepened the understanding of viral pathogenesis., (Copyright © 2023 by The American Association of Immunologists, Inc.)

Porcine deltacoronavirus (PDCoV) cause diarrhea and dehydration in newborn piglets and has the potential for cross-species transmission. Rapid and early diagnosis is important for preventing and controlling infectious disease. In this study, two monoclonal antibodies (mAbs) were generated, which could specifically recognize recombinant PDCoV nucleocapsid (rPDCoV-N) protein. A colloidal gold immunochromatographic assay (GICA) strip using these mAbs was developed to detect PDCoV antigens within 15 min. Results showed that the detection limit of the GICA strip developed in this study was 10 3 TCID 50 /ml for the suspension of virus-infected cell culture and 0.125 μg/ml for rPDCoV-N protein, respectively. Besides, the GICA strip showed high specificity with no cross-reactivity with other porcine pathogenic viruses. Three hundred and twenty-five fecal samples were detected for PDCoV using the GICA strip and reverse transcription-quantitative real-time PCR (RT-qPCR). The coincidence rate of the GICA strip and RT-qPCR was 96.9%. The GICA strip had a diagnostic sensitivity of 88.9% and diagnostic specificity of 98.5%. The specific and efficient detection by the strip provides a convenient, rapid, easy to use and valuable diagnostic tool for PDCoV under laboratory and field conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wang, Fan, Zhang, Guo, Zhao, Zhou, Zhou, Peng, Zhu, Li and Li.)

Objective: Inflammation plays an important role in the pathophysiology of ischemic cardiomyopathy (ICM). We aimed to identify potential biomarkers of inflammation-related genes for ICM and build a model based on the potential biomarkers for the diagnosis of ICM., Materials and Methods: The microarray datasets and RNA-Sequencing datasets of human ICM were downloaded from the Gene Expression Omnibus database. We integrated 8 microarray datasets via the SVA package to screen the differentially expressed genes (DEGs) between ICM and non-failing control samples, then the differentially expressed inflammation-related genes (DEIRGs) were identified. The least absolute shrinkage and selection operator, support vector machine recursive feature elimination, and random forest were utilized to screen the potential diagnostic biomarkers from the DEIRGs. The potential biomarkers were validated in the RNA-Sequencing datasets and the functional experiment of the ICM rat, respectively. A nomogram was established based on the potential biomarkers and evaluated via the area under the receiver operating characteristic curve (AUC), calibration curve, decision curve analysis (DCA), and Clinical impact curve (CIC)., Results: 64 DEGs and 19 DEIRGs were identified, respectively. 5 potential biomarkers (SERPINA3, FCN3, PTN, CD163, and SCUBE2) were ultimately selected. The validation results showed that each of these five potential biomarkers showed good discriminant power for ICM, and their expression trends were consistent with the bioinformatics results. The results of AUC, calibration curve, DCA, and CIC showed that the nomogram demonstrated good performance, calibration, and clinical utility., Conclusion: SERPINA3, FCN3, PTN, CD163, and SCUBE2 were identified as potential biomarkers associated with the inflammatory response to ICM. The proposed nomogram could potentially provide clinicians with a helpful tool to the diagnosis and treatment of ICM from an inflammatory perspective., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Xie, Cheng, Li, Chen and Zhu.)

With the rapid development of synthetic biology, a variety of biopolymers can be obtained by recombinant microorganisms. Polyhydroxyalkanoates (PHA) is one of the most popular one with promising material properties, such as biodegradability and biocompatibility against the petrol-based plastics. This study reviews the recent studies focusing on the microbial synthesis of PHA, including chassis engineering, pathways engineering for various substrates utilization and PHA monomer synthesis, and PHA synthase modification. In particular, advances in metabolic engineering of dominant workhorses, for example Halomonas, Ralstonia eutropha, Escherichia coli and Pseudomonas, with outstanding PHA accumulation capability, were summarized and discussed, providing a full landscape of diverse PHA biosynthesis. Meanwhile, we also introduced the recent efforts focusing on structural analysis and mutagenesis of PHA synthase, which significantly determines the polymerization activity of varied monomer structures and PHA molecular weight. Besides, perspectives and solutions were thus proposed for achieving scale-up PHA of low cost with customized material property in the coming future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gao, Yang, Wang, Xie, Liu, Lin, Han, Zhu, Neureiter, Lin and Ye.)

Avian leukosis virus subgroup J (ALV-J), an oncogenic retrovirus, induces myelocytoma and various other tumors in broilers and layers. Many recent studies have shown that ALV-J can hijack host molecules to facilitate infection. However, the molecular mechanisms of this process are not clear. Here, we aimed to elucidate the molecular mechanisms contributing to ALV-J infection. ALV-J hijacked MIF via p10 and p27 to facilitate ALV-J infection. ALV-J persistently activated MIF expression in DF-1 cells, and MIF significantly facilitated ALV-J internalization and replication, which demonstrated by MIF overexpression and knockdown experiments and treatment with the MIF antagonist ISO-1. Furthermore, we found that the two subunit proteins of Gag, p10 and p27, interacted with MIF in the cytoplasm, respectively. These results suggested that the p10 and p27 subunit in Gag protein recruited MIF to promote ALV-J infection, providing insights into the roles of the p10/p27 and the host factor MIF in ALV-J infection. The finding may facilitate the development of new strategies for controlling ALV-J or retrovirus infections., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: Buyang Huanwu Decoction (BYHW), a famous herbal prescription in traditional Chinese medicine (TCM), has been used for 200 years for treating ischemic heart failure (IHF). This study aims to assess the efficacy and safety of BYHW combined with guideline-guided pharmacotherapy in patients with IHF and explore the biological mechanism by which BYHW exerts its efficacy. Methods: In the multicenter, double-blind, randomized controlled trial, a total of 80 patients with IHF were randomized to receive BYHW or placebo for 3 months. The primary efficacy endpoints were New York Heart Association (NYHA) classification, TCM syndrome scores, N-terminal pro-B-type natriuretic peptide (NT-ProBNP), whereas the mechanism exploration endpoints included energy metabolism parameters and coagulation function parameters. In addition, we performed the proteomic study of the serum of patients after treatment by label-free quantification technology to verify the candidate target proteins and pathways. Results: After 3 months of treatment, the NYHA classification, TCM syndrome scores, and the percentage of subjects with at least 30% reduction in NT-ProBNP were significantly improved in the BYHW group, compared with the control group ( p < 0.05); BYHW treatment also significantly regulated blood glucose, blood lipid levels, ameliorated energy metabolism and improved coagulation function parameters. There were no significant differences in safety endpoints between the two groups. In addition, we obtained 56 differentially expressed proteins by proteomics, including 20 upregulated proteins and 36 downregulated proteins. Bioinformatic analysis revealed the mechanism of BYHW treatment was significantly related to complement and coagulation cascades, cholesterol metabolism, NF-kappa B signaling pathway, PI3K-Akt signaling pathway, and metabolic pathways. Among these differentially regulated proteins, fibrinogen gamma (FGG), fibrinogen beta (FGB), Carboxypeptidase B2 (CPB2), Coagulation factor XIII A (F13A1), Intercellular adhesion molecule1 (ICAM1), Apolipoprotein C-II(APOC2), Apolipoprotein C-I(APOC1), and CD44 were found to be signature proteins associated with the efficacy of BYHW against IHF. Conclusion: BYHW treatment can further improve cardiac dysfunction and clinical symptoms in IHF based on standard therapy without apparent adverse effects. Additionally, BYHW may play a therapeutic role in IHF by improving energy metabolism and regulating coagulation function through multiple targets and pathways., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer MW declared a shared parent affiliation with the author(s) YL, JR, BM, and JL to the handling editor at the time of review., (Copyright © 2022 Zhu, Wei, Li, Wang, Ren, Li, Ma, Wang, Qiao, Zhou and Liu.)

The CCCH-type zinc finger antiviral protein (ZAP) can recognize and induce the degradation of mRNAs and proteins of certain viruses, as well as exerting its antiviral activity by activating T cells. However, the mechanism of ZAP that mediates T cell activation during virus infection remains unclear. Here, we found a potential function of ZAP that relieves immunosuppression of T cell induced by avian leukosis virus subgroup J (ALV-J) via a novel signaling pathway that involves norbin-like protein (NLP), protein kinase C delta (PKC-δ), and nuclear factor of activated T cell (NFAT). Specifically, ZAP expression activated T cells by promoting the dephosphorylation and nuclear translocation of NFAT. Furthermore, knockdown of ZAP weakened the reactivity and antiviral response of T cells. Mechanistically, ZAP reduced PKC-δ activity by upregulating and reactivating NLP by competitively binding with viral protein. Knockdown of NLP decreased the dephosphorylation of PKC-δ by ZAP expression. Moreover, we show that knockdown of PKC-δ reduced the phosphorylation levels of NFAT and enhanced its nuclear translocation. Taken together, these data revealed that ZAP relieves immunosuppression caused by ALV-J and mediates T cell activation through the NLP-PKC-δ-NFAT pathway. IMPORTANCE The evolution of the host defense system is driven synchronously in the process of resisting virus invasion. Accordingly, host innate defense factors effectively work to suppress virus replication. However, it remains unclear whether the host innate defense factors are involved in antiviral immune responses against the invasion of immunosuppressive viruses. Here, we found that CCCH-type zinc finger antiviral protein (ZAP) effectively worked in resistance to immunosuppression caused by avian leukosis virus subgroup J (ALV-J), a classic immunosuppressive virus. Evidence showed that ZAP released the phosphatase activity of NLP inhibited by ALV-J and further activated NFAT by inactivating PKC-δ. This novel molecular mechanism, i.e., ZAP regulation of the antiviral immune response by mediating the NLP-PKC-δ-NFAT pathway, has greatly enriched the understanding of the functions of host innate defense factors and provided important scientific ideas and a theoretical basis for research on immunosuppressive viruses and antiviral immunity.

Objective: Shexiang Baoxin Pill (SBP) is a licensed Chinese herbal pharmaceutical that has been widely accustomed to treat coronary heart disease (CHD) after percutaneous coronary intervention (PCI). This study points to systematically assess the efficacy and security of the combination of SBP with conventional western medicine in the treatment of CHD after PCI., Methods: Databases including PubMed, the Cochrane Library, Web of Science, Embase, CNKI, Wanfang, VIP, and SINOMED were searched to collect RCTs on SBP in CHD after PCI before July 2021. Review Manager 5.3 was used to analyze the data. The Cochrane Collaboration Bias Risk Tool is used to assess the quality of methods., Results: A total of 19 eligible trials of 2022 patients with CHD after PCI were finally included. The results of the aggregate evidence showed that, compared with routine western medicine treatment alone, the combination of SBP with conventional treatment trial groups could significantly reduce the incidence of major adverse cardiac events (MACE) of the patients (RR = 0.38, 95% CI (0.29, 0.51), P < 0.00001). SBP also significantly enhanced left ventricular ejection fraction (LVEF) (MD = 4.00, 95% CI (3.42, 4.58), P < 0.00001) and lessened N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) levels (MD = -167.18, 95% CI (-174.80, -159.57), P < 0.00001). In addition, the inflammatory mediators and blood lipid index in the experimental group after the combined therapy were also mediated ( P < 0.05). Moreover, SBP did not increase the incidence of adverse reactions during treatment. The results of subgroup analysis illustrated that the length of the intervention course might be the source of the heterogeneity of NT-pro-BNP and hs-CRP., Conclusion: SBP could demonstrate a beneficial role in patients with CHD after PCI of reducing the incidence of MACE and improving LVEF, NT-pro-BNP, inflammatory mediators, and blood lipid index. However, limited by the quantity and quality of eligible studies, the above conclusions required more standardized, rigorous, high-quality clinical trials to verify further., Competing Interests: All authors of this study claim there is no conflict of interest., (Copyright © 2021 Jingjing Wei et al.)

Objective: To predict the main active ingredients, potential targets, and key pathways of Jiawei Chaiqin Wendan decoction treatment in vestibular migraine and explore possible mechanisms by network pharmacology and molecular docking technology., Methods: The active ingredients and related targets of Jiawei Chaiqin Wendan decoction were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The corresponding genes of the target were queried by UniProt database, and the "drug-compound-target-disease" network was constructed by Cytoscape 3.7.2 software. GO functional enrichment analysis and KEGG pathway enrichment analysis were carried out by R software and Bioconductor, and column chart and bubble chart were drawn by Prism software and OmicShare database for visualization. Finally, the mechanism and potential targets of Jiawei Chaiqin Wendan decoction in the treatment of vestibular migraine were predicted., Results: The "drug-compound-target-disease" network contains 154 active ingredients and 85 intersection targets. The key targets include AKT1, IL6, MAPK3, VEGFA, EGFR, CASP3, EGF, MAPK1, PTGS2, and ESR1. A total of 1939 items were obtained by GO functional enrichment analysis ( P  < 0.05). KEGG pathway enrichment analysis screened 156 signal pathways ( P  < 0.05), involving PI3K-Akt signal pathway, AGE-RAGE signal pathway in diabetes complications, MAPK signal pathway, HIF-1 signal pathway, IL-17 signal pathway, etc. Molecular docking results showed that quercetin, luteolin, kaempferol, tanshinone IIa, wogonin, naringenin, nobiletin, dihydrotanshinlactone, beta-sitosterol, and salviolone have good affinity with core target proteins IL6, PTGS2, MAPK1, MAPK3, and CGRP1., Conclusion: The active ingredients in Jiawei Chaiqin Wendan decoction may regulate the levels of inflammatory factors and neurotransmitters by acting on multiple targets such as IL6, MAPK3, MAPK1, and PTGS2, so as to play a therapeutic role in vestibular migraine., Competing Interests: The authors declare no conflicts of interest regarding the publication of this article., (Copyright © 2021 Menglin Liu et al.)

The effects of mulberry polyphenols and malondialdehyde (MDA) on the emulsifying and gel properties of myofibrillar protein (MP) were studied. Emulsibility and gel properties of MP were compared in range of mulberry polyphenol/MDA concentrations by particle size, Zeta potential, antioxidant capacity, gel strength, water-holding capacity (WHC), rheological properties and microstructure. Mulberry polyphenols enhanced the inoxidizability of MP emulsion but decreased its emulsifying property. MDA at intermediate concentrations (5-20 mM) improved the elasticity, strength, and WHC of polyphenols-modified MP emulsion gel, while at high concentration (40 mM) it destroyed the emulsion gel, resulting in "oil leakage". Polyphenol is not conducive to the gelation but weaken the oxidative damage of MDA to protein. The gel structure of MP emulsion collapsed after high dose of polyphenols or MDA treated. Thus, to maintain uniform textural and antioxidant activity of meat product, both polyphenols addition and oxidation intensity should be controlled simultaneously., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The effects of rutin, quercetin, and caffeic acid on protein oxidation in Cantonese sausage during 60 days of storage at room temperature (25 ± 1 °C) were investigated. The results showed that the three phenolic compounds played different roles in inhibiting the oxidation of sarcoplasmic and myofibrillar protein. All of them inhibited sarcoplasmic protein oxidation by retarding carbonylation, the conversion of SH to S-S groups, and the formation of dimeric tyrosine and Schiff bases, of which rutin is the most effective. For myofibrillar protein, all of them suppressed the conversion of SH to S-S groups, only caffeic acid inhibited the accumulation of Schiff bases instead of carbonyls while both quercetin and caffeic acid inhibited the formation of dimeric tyrosine. In addition, quercetin had an inverse dosage effect on the oxidation regulation of MP, 0.16 g/kg quercetin had better inhibit effect on protein oxidation than 0.32 g/kg quercetin., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

To systematically evaluate the efficacy and safety of sofren injection combined with conventional Western medicine in the treatment of angina pectoris. Randomized controlled trials (RCTs) on the treatment of angina pectoris with sofren injection combined with Western medicine were collected by searching PubMed, the Cochrane Library, Embase, Web of Science, CNKI, Wanfang Database, Weipu Database, and China Biomedical Literature Service System (CBM) by computer with the retrieval time from establishment of database to August 2020. After literature screening according to the predetermined inclusion and exclusion criteria, data of eligible studies were extracted, and then, a meta-analysis was conducted with the RevMan 5.3 software. The results of meta-analysis showed that the combination of sofren injection and Western medicine improved the platelet aggregation rate of patients (MD = -5.53, 95% CI (-6.42, -4.64), P < 0.00001), PAI-1 (SMD = -2.29, 95% CI (-2.57, -2.01), P < 0.00001), TXB2 (MD = -11.91, 95% CI (-14.50, -9.32), P < 0.00001), duration of angina attack (MD = -2.01, 95% CI (-3.14, -0.87), P =0.0005), ECG symptoms (RR = 1.29, 95% CI (1.20, 1.37), P < 0.00001), whole blood viscosity (MD = -1.07, 95% CI (-1.66, -0.48), P =0.0004), plasma viscosity (MD = -0.27, 95% CI (-0.35, -0.20), P < 0.00001), fibrinogen (MD = -0.67, 95% CI (-0.84, -0.50), P < 0.00001), whole blood high shear viscosity (MD = -1.04, 95% CI (-1.30, -0.79), P < 0.00001), whole blood low shear viscosity (MD = -2.03, 95% CI (-2.53, -1.53), P < 0.00001), CRP (MD = -1.96, 95% CI (-3.01, -0.91), P =0.0003), IL-6 (MD = -2.79, 95% CI (-4.02, -1.55), P < 0.00001), and TNF- α (MD = -17.34, 95% CI (-25.86, -8.81), P < 0.00001) and better than the Western medicine group, and there was no statistical significance in the incidence of adverse reactions between the two groups ( P =0.48). The clinical application of sofren injection combined with conventional Western medicine in the treatment of angina pectoris is clear and safe, so it is recommended for clinical application., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Genhao Fan et al.)

With the aim of improving the photocatalytic activity for water splitting, novel core-shell-structured crystalline-BaTiO 3 /amorphous-BaTiO 3-x /crystalline-CdS composite nanocubes are prepared by a facile two-step synthesis approach. Basic characterization techniques such as X-ray diffraction, scanning electron microscopy, energy dispersive X-ray spectroscopy and transmission electron microscopy are carried out on the as-prepared composite nanocubes in order to confirm the quality of their crystal structure, morphology and chemical components correspondingly. UV-Vis-NIR measurements of the as-prepared composite nanocubes validate the presence of extended visible-light absorbance due to oxygen-deficient BaTiO 3-x . Photoelectrochemical tests are carried out on the as-prepared nanocomposite films that are coated directly on indium tin oxide (ITO) glass substrates. The as-prepared composite nanocubes show a photocurrent density of 100 μA cm -2 without electric field poling, whereas they show about 200 μA cm -2 with an electric field poling of 18.8 kV cm -1 . This study suggests that the photoelectrochemical performance is highest in our prepared BaTiO 3 /BaTiO 3-x /CdS composite film compared to the pure BaTiO 3 , CdS and BaTiO 3 /BaTiO 3-x films, and it may offer a new potential route for designing cost-effective, highly stable and efficient photocatalysts.

Shenfuyixin granule (SFYXG, i.e., Xinshuaikang granule) is a prescription, commonly used in the clinical experience, which plays a significant role in the treatment of heart failure. The purpose of this present research was to investigate the protective effect of SFYXG, and the mechanism about anti-H 2 O 2 -induced oxidative stress and apoptosis in the neonatal rat cardiomyocytes. Myocardial cells, as is well known, were divided into 4 groups: normal, model, SFYXG, and coenzyme Q10 group, respectively. Cells viability was determined by MTT assay. Flow cytometry and AO/EB staining were implemented to test the apoptosis rate and intracellular reactive oxygen species (ROS) level. Mitochondrion membrane potential (MMP) was evaluated by JC-1 fluorescence probe method. The myocardial ultrastructure of mitochondrion was measured by electron microscope. The related mRNA expression levels of Bax, Bcl-2, Caspase-3, caspase-8, and caspase-9 were detected by real-time polymerase chain reaction (PCR). Also, the expression levels of Bax and Bcl-2 protein were detected by Western blot, and the expression levels of caspase-3, caspase-8, and caspase-9 protein were tested by caspase-Glo®3 Assay, caspase-Glo®8 Assay, and caspase-Glo®9 Assay, respectively. GAPDH was used as the internal reference gene/protein. The results revealed that SFYXG (0.5 mg/ml) raised the viability of myocardial cell, weakened the apoptosis rate and ROS level, corrected the mitochondrion membrane potential stability, and improved cell morphology and ultrastructure of myocardial mitochondrion. Furthermore, SFYXG upregulated the antiapoptosis gene of Bcl-2, but downregulated the proapoptosis genes of Bax, caspase-3, and caspase-9. In conclusion, SFYXG could appear to attenuate myocardial injury by its antioxidative and antiapoptosis effect., Competing Interests: The authors have no potential conflicts of interest to disclose., (Copyright © 2021 Xinlu Wang et al.)

Background: Coronary heart disease (CHD) has become a common cardiovascular disease that seriously threatens the health of people. As reperfusion in the early phase and drug therapy, especially percutaneous coronary intervention (PCI), have become widely used in the clinic, the mortality of acute myocardial infarction in the short term has been reduced significantly. In addition, in 40%-56% of patients who experience myocardial infarction, cardiac dysfunction occurs and about 25%-33% develop heart failure., Methods: This study was designed as a multicenter, double-blind, randomized, placebo-controlled, parallel-group, superiority trial. Participants were randomly assigned in a 1:1 ratio through a centrally controlled, computer-generated, simple randomization schedule. The primary outcome was left ventricular end-diastolic volume index = left ventricular end-diastolic volume/body surface area. The combined secondary outcomes include traditional Chinese medicine syndrome score, echocardiogram results, 6-minute walk test results, Seattle Angina Questionnaire score, cardiac magnetic resonance imaging results, biological indicators, dynamic electrocardiogram results, and experiment event rate. Assessments will be performed at baseline and at 4, 8, and 12 weeks after randomization., Discussion: This trial will demonstrate that the addition of a Tongmai Yangxin pill (TMYX) to conventional treatment will intervene in the development of cardiac remodeling and cardiac dysfunction., Trial Registration: This study was registered in the Chinese Clinical Trial Registry on 7 May 2019. The registration number is ChiCRT1900023023 (http://www.chictr.org.cn/showproj.aspx?proj=12370).

A fully automatic system, which integrated cross used solid-phase extraction with ultra-high performance liquid chromatography-tandem mass spectrometry, was developed and validated for the simultaneous determination of multi-class pharmaceuticals (62 in total) in Milli-Q water, tap water, lake water, and ground water. The online system allowed the cross-utilization of two SPE columns without significant carryover and achieved an automatic, sensitive and fast analysis, requiring about 14 min per analysis. The features of the online system were systematically investigated and the analytical conditions were fully optimized. Sixty-two pharmaceuticals were divided into two groups (acidic and basic) under different extraction conditions to increase the extraction efficiency. Under optimal conditions, all the correlation coefficients were greater than 0.9929. The LODs and the LOQs were in the range of 0.00119-0.623 ng L -1 and 0.00475-2.49 ng L -1 , respectively. The RSDs% for the intra-/inter-day precision were less than 10.6% and 15.6%, respectively. The system recoveries ranged from 80.7 to 119.9%. Compared with the offline SPE method, the online cross used SPE-UHPLC-MS/MS method obtained higher sensitivity and reduced manual operations. Compared with the existing online SPE systems, this system can reduce the time per analysis. Finally, this online system was applied to the analyses of three real water samples. Based on the results, the online cross used SPE-UHPLC-MS/MS system as an automatic, sensitive and efficient technique showed great promise for the future in the trace analysis of multi-class pharmaceuticals in complex aqueous samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The CCCH-type zinc finger antiviral protein (ZAP), a host antiviral factor, plays an important role in innate defenses. Although the anti-viral mechanism of ZAP has been elucidated, however, the tissue specificity and the viral infection correlativity have not been fully understood. Here, we tested the dynamic distribution and localization of chicken ZAP (chZAP) before and after avian leukosis virus subgroup J (ALV-J) infection. The results showed that chZAP was highly expressed in adrenal gland and testis before ALV-J infection, and significantly upregulated in liver, kidney and bursa of Fabricius, and extremely overexpressed in spleen after ALV-J infection. The results indicated that chZAP is an inducible protein and showed specific overexpression in spleen after ALV-J infection. Furthermore, we demonstrated that chZAP, as a host intracytoplasmic factor, accumulated and migrated to the periphery of nucleus in DF-1 cells post-infection with ALV-J. Taken together, chZAP characterized as an inducible antiviral protein and specifically overexpressed in spleen after ALV-J infection., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The pathogenesis of immunological tolerance caused by avian leukosis virus subgroup J (ALV-J), an oncogenic retrovirus, is largely unknown., Results: In this study, the development, differentiation, and immunological capability of B cells and their progenitors infected with ALV-J were studied both morphologically and functionally by using a model of ALV-J congenital infection. Compared with posthatch infection, congenital infection of ALV-J resulted in severe immunological tolerance, which was identified as the absence of detectable specific antivirus antibodies. In congenitally infected chickens, immune organs, particularly the bursa of Fabricius, were poorly developed. Moreover, IgM-and IgG-positive cells and total immunoglobulin levels were significantly decreased in these chickens. Large numbers of bursa follicles with no differentiation into cortex and medulla indicated that B cell development was arrested at the early stage. Flow cytometry analysis further confirmed that ALV-J blocked the differentiation of CD117 + chB6 + B cell progenitors in the bursa of Fabricius. Furthermore, both the humoral immunity and the immunological capability of B cells and their progenitors were significantly suppressed, as assessed by (a) the antibody titres against sheep red blood cells and the Marek's disease virus attenuated serotype 1 vaccine; (b) the proliferative response of B cells against thymus-independent antigen lipopolysaccharide (LPS) in the spleen germinal centres; and (c) the capacities for proliferation, differentiation and immunoglobulin gene class-switch recombination of B cell progenitors in response to LPS and interleukin-4(IL-4) in vitro., Conclusions: These findings suggested that the anergy of B cells in congenitally infected chickens is caused by the developmental arrest and dysfunction of B cell progenitors, which is an important factor for the immunological tolerance induced by ALV-J.

Background: Stable angina is a common cardiovascular disease with high mortality and a poor prognosis. Although there are various interventions against stable angina, none are able to significantly reduce the mortality rate. Guanxinjing capsule (GXJ) is made from the classical Chinese prescription Xuefuzhuyutang (). Both basic research and clinical studies have shown that GXJ can relieve the symptoms of angina, but currently, the effects of GXJ lack high-quality clinical evidence. The aim of this study was to evaluate the clinical effectiveness and safety of GXJ compared with placebo., Methods/design: This multicentre, blinded, randomized trial will be conducted with a total of 120 participants diagnosed with chronic stable angina (Qi deficiency and blood stasis syndrome). Using a central randomization system, participants will be randomized (1:1) into groups receiving either GXJ or placebo for 8 weeks. After a 2-week run-in period, eligible patients will receive either GXJ or placebo (4 pills, three times daily) for 8 weeks in addition to conventional treatment. The primary outcomes include changes in the total exercise time on exercise tolerance tests and changes in the integral scores of angina symptoms. The secondary outcome measures include changes in the maximal estimated workload, changes in time to a 1 mm ST-segment depression or raise, changes in the time to onset of angina during exercise tolerance testing, changes in the total score of traditional Chinese medicine syndrome, and changes in the total score of the Generalized Anxiety Disorder 7-item assessment between baseline and week 8. Other outcome measures will also be assessed. All exercise tolerance tests use a standard Bruce multistage exercise test protocol. Adverse events will be monitored throughout the trial., Discussion: This study will investigate whether GXJ can alleviate clinical symptoms, increase the angina-free walking time, and improve quality of life in patients with chronic stable angina (Qi deficiency and blood stasis syndrome). The results of this study will provide clinical evidence for the application of GXJ to the treatment of stable angina., Trial Registration: Chinese Clinical Trial Registry, ChiCTR1800014258 . Registered on 2 January 2018.

Background: Traditional Chinese medicine injections (TCMIs) have been widely used to treat severe and acute diseases due to their high bioavailability, accurate curative effect, and rapid effect. However, incidence rates of adverse drug reactions (ADRs) of TCMIs have also increased in recent years. Xueshuantong injection (XSTI) is a commonly-used TCMI comprised of Panax notoginseng total sapiens for the treatment of stroke hemiplegia, chest pain, and central retinal vein occlusion. Its safety remains uncelar. Therefore, post-marketing safety of XSTI was studied in this research., Methods: In present study, post-marketing safety surveillance and re-evaluation of XSTI were reported. Thirty thousand eight hundred eighty-four patients in 33 hospitals from 7 provinces participated in this study. Incidence rate, most common clinical manifestations, types, severity, occurrence time, and disposal of ADRs were calculated., Results: Incidence rate of ADR of XSTI was 4.14‰ and the most common clinical manifestations were skin and its appendages damage. Type A accounts for 95.49% of ADRs of XSTI and most of them (41.41%) were occurred within 24 h after receiving XSTI treatment. Severities of most ADRs of XSTI were moderate reactions (86.72%). Main disposition of ADRs of XSTI was drug withdrawal and symptomatic treatment (54.69%)., Conclusions: Our data provide basis for improvement of instructions of XSTI and clinical safety of XSTI. Post-marketing surveillance of TCMIs in this study is a powerful tool to identify types and manifestations of ADRs to improve safety and effectiveness of drugs in clinical applications., Trial Registration: This protocol has international registration in China clinical trial registration center ( ChiCTR~OPC~ 14,005,718 ) at December 22, 2014.

Background and Aims: Transient receptor potential vanilloid type 1 channel (TRPV1) is found to be expressed in endothelial cells (ECs) and activate endothelial nitric oxide synthase (eNOS). Recent studies implicate TRPV1 in attenuating inflammatory responses. However, the mechanisms underlying the beneficial effects remain unclear. In this study, we investigated whether TRPV1 suppresses inflammatory responses of ECs via eNOS/NO pathway., Methods: Human umbilical vein endothelial cells (HUVECs) and renal microvascular endothelial cells (MVECs) isolated from deoxycorticosterone (DOCA)-salt hypertensive mice were cultured in the presence of capsaicin (CAP, a specific TRPV1 agonist) with or without the specific inhibitor of TRPV1, NOS, or Ca 2+ -dependent phosphatidylinositol 3-kinase (PI3K)/Akt pathway, before lipopolysaccharide (LPS) stimulation. NO metabolites, protein expression, and inflammatory molecules were evaluated by Griess assay and immune assay-based multiplex analysis, respectively. Monocyte adhesion was determined by measuring the fluorescently labeled human monocytes attached to LPS-stimulated ECs., Results: In HUVECs, treatment with CAP increased NO production, and CAP-induced NO production was accompanied by increased eNOS ser1177 phosphorylation. Additionally, CAP attenuated LPS-induced cytokine and chemokine production, adhesion molecule expression, activation of NF-κB, and monocyte adhesion in HUVECs, and these effects were abrogated by the inhibition of TRPV1, NOS, or Ca 2+ -dependent PI3K/Akt pathway. Moreover, these protective actions of TRPV1 were also observed in renal MVECs isolated from DOCA-salt hypertensive mice., Conclusions: Our results indicate that TRPV1 activation suppresses the inflammatory response of ECs via the activation of Ca 2+ /PI3K/Akt/eNOS/NO pathway, the protective effects are also documented in ECs derived from salt-sensitive hypertensive mice., (Copyright © 2017. Published by Elsevier B.V.)

Although current vaccination strategies have been successful at preventing a variety of human diseases, attempts at vaccinating against some pathogens such as AIDS and tuberculosis (TB) have been more problematic, largely in that abnormally high numbers of antigen specific CD8+ T cells are required for protection. This study assessed the effect of temporarily dampening the chemokine receptor CXCR3 and CCR5 after vaccination on host immune responses by the administration of TAK-779, a small molecule CXCR3 and CCR5 antagonists commonly used to inhibit HIV infection. Our results showed that the use of TAK-779 enhanced memory CD8+ T cell immune responses both qualitatively and quantitatively. Treatment with TAK-779 following vaccination of an influenza virus antigen resulted in enhanced memory generation with more CD8+CD127+ memory precursor and fewer terminally differentiated effector CD8+CD69+ T cells. These memory T cells were able to become IFN-γ-secreting effector cells when re-encountered the same antigen, which can further enhance the efficacy of vaccination. The mice vaccinated in the presence of TAK-779 were better protected upon influenza virus challenge than the control. These results showed that vaccination while temporarily inhibiting chemokine receptor CXCR3 and CCR5 by TAK-779 could be a promising strategy to generate large number of protective memory CD8+ T cells.

A novel co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense with pretreated sugarcane bagasse (SCB) under mild alkali conditions for bio-hydrogen production was established, exhibiting a cost-effective and synergetic advantage in bio-hydrogen production over monoculture of C. thermocellum or T. aotearoense with untreated SCB. The optimized pretreatment conditions were established to be 3% NaOH, and a liquid to solid ratio of 25:1 at 80°C for 3h. A final hydrogen production of 50.05±1.51 mmol/L was achieved with 40 g/L pretreated SCB at 55°C. The established co-culture system provides a novel consolidated bio-processing strategy for bioconversion of SCB to bio-hydrogen., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

An efficient pretreatment method using NH(4)OH-H(2)O(2) and ionic liquid (IL) was developed for the recovery of cellulose from sugarcane bagasse (SCB). The regenerated SCB from the combined pretreatment exhibited significantly enhanced enzymatic digestibility with an efficiency of 91.4% after 12h of hydrolysis, which was 64% higher than the efficiency observed for the regenerated SCB after the individual NH(4)OH-H(2)O(2) pretreatment. 1-Allyl-3-methylimidazolium chloride ([Amim]Cl) dissolved the cellulose from the NH(4)OH-H(2)O(2)-pretreated SCB, and the crystallinity index (CrI) detected by X-ray diffraction (XRD) was reduced by 42%. The recycled and fresh [Amim]Cl demonstrated the same performance on the pretreatment of SCB for the enhancement of enzymatic digestibility. The regenerated SCB was subsequently used in simultaneous saccharification and co-fermentation (SScF) for bioethanol production by cellulase and yeast. The pretreatment did not have a negative effect on bioethanol fermentation, and an ethanol yield of 0.42 g/g was achieved with a corresponding fermentation efficiency of 94.5%., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A biphasic fermentation approach was undertaken for the production of ethanol and hydrogen from cassava pulp. The glucose generated by co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense was 13.65±0.45 g L(-1), which was 1.75 and 1.17-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. The accumulated glucose could be utilised rapidly by subsequently inoculated Saccharomyces cerevisiae. An inoculum ratio of 1:1, a thermophilic fermentation of 84 h, and a pulp concentration of 4% proved optimal for ethanol production, fermentation efficiency, and productivity. With these conditions, the ethanol level reached 8.83±0.31 g L(-1) with a fermentation efficiency of 64.95±2.71%. Hydrogen production of 4.06 mmol by the co-culture system was 1.54 and 2.09-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. This sequential co-culture approach provided a consolidated bio-processing means to produce ethanol and hydrogen from cassava pulp., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

To enhance ethanol production in Thermoanaerobacterium aotearoense, the lactate dehydrogenase (ldh) gene, which is responsible for lactic acid production in a key branch pathway, was successfully disrupted via homologous recombination. ldh-up and ldh-down were designed and amplified based on JW/SL-YS485-AY 278026, and they were subsequently used as homologous fragments with an inserted erythromycin resistance gene to construct the targeted vector based on pBLUESCRIPT II SK(+). Southern hybridization and PCR-based assay definitely confirmed that the ldh gene in the Δldh mutant was disrupted by the insertion of the erythromycin resistance gene. Compared with the wild type, the Δldh mutant exhibited increases of 31.0% and 31.4% in cell yield under glucose and xylose cultivation, respectively, probably because knocking out the ldh gene results in increased acetate and ATP levels. Knockout of lactate dehydrogenase produced 2.37- and 2.1-fold increases in the yield of ethanol (mole/mole substrate) under glucose and xylose cultivation, respectively. Moreover, no lactic acid was detected in Δldh mutant fermentation mixtures (detection limit of HPLC: 0.5 mM), but lactic acid was readily detected for growth of the wild-type strain on both glucose and xylose, with final concentrations up to 59.24 mM and 56.06 mM, respectively. The success of this process thoroughly demonstrates the methodological possibility of gene knockout through homologous recombination in Thermoanaerobacterium., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: To study the changes of vitamin nutritional status of residents in a high-risk area of esophageal cancer., Methods: 8 towns where nutrition survey had been done were selected, 2 villages were randomly selected from each town and 30 families were randomly selected per village. The blood and 4h burden urine samples were collected from subjects. The serum vitamin A, blood total ascorbic acid and glutathione reductase activition coefficient (AC) in red blood cell were determinde by using of trifluoroacetic acid colorimetry, 2, 4-dinitrophenylhydrazine colorimetric method and Sauberlich method respectively. The contents of vitamin B2 , reduction ascorbic acid and total ascorbic acid in 4h burden urine were measured by using of fluorescent pectrophotometry,2,6-Dichloro-N-(4-hydroxyphenyl)-1,4-benzoquinoneimine titrimetric method and 2,4-dinitrophenylhydrazine colorimetric method respectively. The results were analyzed and compared with those of in 1980, 1983 and 1984. The t-test or chi2-test was carried out., Results: The level of serum retinal in residents living in Ren cun, Dongyao and Hejian is significantly higher than that of in 1982 (P < 0.01). The level of vitamin B2 in 4h burden urine in residents living in Yaocun, Rencun and Heshun are significantly higher than that of in 1982 or 1984 (P < 0.01). The number of ribflavin deficiency of population living in Dongyao, Hejian, Yaocun and Rencun are significantly lower than that of in 1982 or 1983 (P < 0.01) according to the content of 4h burden urine and glutathione reductase activation coefficient (AC) in red blood cell; the number of vitamin C deficiency of population living in Dongyao, Hejian,Yao cun, Ren cun and Heshun are significantly lower than that of in 1982 or 1983 (P < 0.01) according to blood total ascorbic acid and the ascorbic acid content of 4h-burden urine and., Conclusion: Vitamin nutritional status of residents in a high-risk area of esophageal cancer have been improved.