Tropical Medicine and Infectious Disease
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2023
·Vol. 8
·期 2
·105
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IF 2.9
Fengyu Tian,Cong Jin,Shangzhi Ji,Yanqing Tie,Guohao Fan,Ruiqing Zhang,Yehuan Zheng,Xinxin Shen,Xuejun Ma,Zhishan Feng
Human immunodeficiency virus 1 (HIV-1) attacks the immune system, making people susceptible to various diseases, thus increasing their risk of death. Comprehensive detection of major HIV-1 strains circulating in China is vital for effective HIV-1 infection prevention and treatment. HIV-1 nucleic acid detection is considered effective for HIV-1 diagnosis since traditional immunological testing may fail to detect HIV-1 infection during the window period. This work demonstrates a one-pot two-stage amplification assay (RT-RAP), a combination of reverse transcription recombinase (RT- RAA), and quantitative real-time polymerase chain reaction (qRT-PCR). The turn-around time of the assay is only 50 min and can be performed with commonly available laboratory equipment, the qPCR devices. The RT-RAP assay could detect approximately 5 and 14 copies/reaction of HIV-1 DNA and RNA using recombinant plasmids and standard reference strains, respectively. Additionally, we found that the clinical performance of RT-RAP (detected 169 samples out of 170 specimens) was consistent with that of qRT-PCR. The sensitivity and specificity of RT-RAP were 100.00% (99/99) and 98.59% (70/71), respectively, while its positive and negative predictive values were 99.00% (99/100) and 100.00% (70/70), respectively. The total coincidence rate of the RT-RAP was 99.41% (169/170), with a kappa value of 0.988 (p< 0.05). We demonstrated that RT-RAP could rapidly detect the common HIV-1 subtypes commonly circulating in China with comparable sensitivity and specificity to qRT-PCR.Keywords:HIV-1;RT-RAA;qRT-PCR;clinical detection
研究领域:
病毒学
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分子生物学
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临床诊断
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DOI
PubMed
高影响因子
Frontiers in Plant Science
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2023
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IF 5.6
Guohui Li,Muhammad Aamir Manzoor,Guoyu Wang,Cunwu Chen,Cheng Song
Introduction KNOX plays a pivotal role in governing plant growth, development, and responses to diverse abiotic and biotic stresses. However, information on the relationship between the KNOX gene family and expression levels under different treatments in Dendrobium is still limited. Methods To address this problem, we first used bioinformatics methods and revealed the presence of 19 KNOX genes distributed among 13 chromosomes in the Dendrobium huoshanense genome. Through an analysis of phylogenetic relationships, these genes were classified into three distinct clades: class I, class II, and class M. Our investigation included promoter analysis, revealing various cis -acting elements associated with hormones, growth and development, and abiotic stress responses. Additionally, qRT-PCR experiments were conducted to assess the expression patterns of DhKNOX genes under different treatments, including ABA, MeJA, SA, and drought. Results The results demonstrated differential expression of DhKNOX genes in response to these treatments, thereby highlighting their potential roles in stress adaptation. Discussion Overall, our results contribute important insights for further investigations into the functional characterization of the Dendrobium KNOX gene family, shedding light on their roles in plant development and stress responses.
研究领域:
植物学
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分子生物学
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基因组学
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PubMed
高影响因子
Agronomy-Basel
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2023
·Vol. 13
·期 6
·1489
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IF 3.7
Yidan Kou,Jinpeng Ren,Yujie Ma,Rongrong Guo,Juane Shang,Dongliang Qiu,Cuilan Ma
In this study, the two peach cultivars ‘Baifeng’ and ‘Weiduanmihong’ were used as experimental materials, and their fruits were sprayed with different concentrations of L-glutamic acid, brassinolide, and sucrose to study the effects of these three exogenous substances on fruit quality and anthocyanin metabolism of peaches. The results showed that the appearance quality (average single fruit weight, fruit firmness, and peel color difference), nutritional quality (soluble solids, soluble sugar, titratable acid, anthocyanins, total phenols, flavonoids, etc.), peel anthocyanin-related enzyme activity, and related gene expression of ‘Baifeng’ and ‘Weiduanmihong’ peaches treated with three different exogenous substances were different from those of the control. Higher-concentration treatments could significantly improve the appearance of peach fruit, the nutritional quality of peach fruit and the activity of anthocyanin-related enzymes in peel, as well as promote the expression of related genes. Treatment with 400 mg/L L-glutamic acid significantly promoted the average fruit weight of ‘Baifeng’ peaches. Treatment with 800 mg/L L-glutamic acid significantly promoted the increase in PAL enzyme activity and the expression ofPpPALandPpF3Hin the two peach varieties and significantly promoted the expression of anthocyanin metabolism genesPpF3′HandPpGST1in ‘Baifeng’ peach peel and anthocyanin metabolism genePpUFGTin ‘Weiduanmihong’ peach peel. Treatment with 34 mg/L sucrose significantly increased the fruit firmness of ‘Baifeng’ peaches and the soluble sugar content of ‘Weiduanmihong’ peaches. Treatment with 51 mg/L sucrose significantly promoted the increase in flavonoid content andPpUFGTexpression in ‘Baifeng’ peach fruit and significantly promoted the expression of anthocyanin metabolism genesPpDFRandPpANSin ‘Weiduanmihong’ peach peel. Treatment with 0.6 mg/L brassinolide significantly promoted the increase in soluble solids (TTS), soluble sugar, anthocyanin, total phenol content, PAL enzyme activity, UFGT enzyme activity, and the expression of anthocyanin metabolism genesPpDFRandPpMYB10.1in ‘Baifeng’ peach fruit, and it significantly increased the average single fruit weight, fruit hardness, anthocyanin content, and UFGT enzyme activity of ‘Weiduanmihong’ peach fruit and promoted the expression of anthocyanin metabolism genesPpF3HandPpGST1in ‘Weiduanmihong’ peach peel. The comprehensive effect of 0.6 mg/L brassinolide treatment on improving peach fruit quality and increasing anthocyanin content produces the best results and could be popularized in production practices.Keywords:peach (Prunus persicaL.);exogenous substance treatment;fruit quality;anthocyanin;qRT-PCR technology
研究领域:
农学
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植物生理学
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分子生物学
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高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2021
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IF 3.963
Meng-Wen Yan,Xiao-Rong Xing,Fu-An Wu,Jun Wang,Sheng Sheng
Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae) is a predominant endoparasitoid of lepidopteran pests in mulberry fields. Extensive application of insecticides puts natural enemies under threat. UDP-glycosyltransferases (UGTs), as important detoxification enzymes , potentially contribute to the detoxification of pesticides in insects. To investigate the roles of UGTs in the process of tolerance towards commonly used insecticides in M. pulchricornis , ten UGT genes were identified from the transcriptome database of M. pulchricornis . Seven UGT genes contained full-length ORFs and shared 47.12–78.28% identity with other homologous hymenopteran insects. qRT-PCR validation revealed that UGT genes can be induced by treatment of sublethal doses of phoxim, cypermethrin and chlorfenapyr, respectively, and these upregulations were depending on the time post insecticide treatments. To further explore the functions of UGT genes, three MpulUGT genes were singly knocked down, which resulted in the decline of UGT expression and significantly increased mortality of parasitoids under sublethal doses of insecticides exposure. This study revealed that UGTs in M. pulchricornis contributed to the tolerance towards insecticides and provided basic insight into the insecticide detoxification mechanism in parasitoid wasps .
研究领域:
昆虫学
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分子生物学
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害虫管理
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PubMed
高影响因子
Foods
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2021
·Vol. 10
·期 5
·1073
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IF 4.35
Meng-Bo Tian,Lin Yuan,Ming-Yuan Zheng,Zhu-Mei Xi
Anthocyanins are vital components of plant secondary metabolites, and are also the most important coloring substances in wine. Teinturier cultivars are rich in anthocyanins. However, the differences in anthocyanin accumulation and profiles between teinturier and non-teinturier cultivars have not been reported. In this study, Yan 73 and Dunkelfelder were selected as the experimental materials, and three non-teinturier cultivars were used for comparison. LC-MS and qRT-PCR were used to determine the individual anthocyanin contents and the relative gene expression. The results show that the total anthocyanin content of the teinturier cultivars was considerably higher than that in non-teinturier cultivars, and the levels of individual anthocyanins increased gradually during ripening. Lower ratios of modified anthocyanins were found in the teinturier cultivars, which was not only due to the high expression level ofVvUFGTandVvGST4, but also due to the relatively low expression ofVvOMTin these cultivars. Cluster analysis of gene expression and anthocyanin accumulation showed thatVvUFGTis related to anthocyanin accumulation, and thatAM1is related to the synthesis and transport of methylated anthocyanins. Our results will be useful for further clarifying the pathways of anthocyanin synthesis, modification, and transport in teinturier cultivars.Keywords:grape;Yan 73;Dunkelfelder;anthocyanin;cluster analysis
研究领域:
植物学
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植物生理学
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分子生物学
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PubMed
高影响因子
Drug Design Development and Therapy
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2022
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IF 4.319
Xiangtian Ling,Xinqi Ma,Xielan Kuang,Yuxiu Zou,Han Zhang,Han Tang,Han Du,Binbin Zhu,Hao Huang,Qing Xia,Minghao Chen,Danyi Mao,Dongli Chen,Huangxuan Shen,Jianhua Yan
Purpose To assess the cellular and molecular effects of lidocaine on muscles/myoblasts.Methods Cultured myogenic precursor (C2C12) cells were treated with varying concentrations of lidocaine.Results Cell viability of C2C12 cells was inhibited by lidocaine in a concentration-dependent manner, with concentrations ≥0.08%, producing a dramatic reduction in cell viability. These ≥0.08% concentrations of lidocaine arrested cell cycles of C2C12 cells in the G0/G1 phase. Moreover, lidocaine inhibited cell migration and myogenic processes in C2C12 cells at low concentrations. Results from QRT-PCR assays revealed that following treatment with lidocaine, Notch1, Notch2, Hes1, Csl and Dll4 all showed higher levels of expression, while no changes were observed in Mmal1, Hey1, Dll1 and Jag1.Conclusion This work provides the first description of the effects of lidocaine upon the regeneration of muscles and maintenance of satellite cells at the cellular and molecular levels. In specific, we found that the Dll4-Notch-Csl-Hes1 axis was up-regulated suggesting that the Notch signaling pathway was involved in producing these effects of lidocaine. These findings provide a new and important foundation for future investigations into the effects of drug therapies in muscle diseases.
研究领域:
药理学
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细胞生物学
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分子生物学
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PubMed
高影响因子
MARINE ENVIRONMENTAL RESEARCH
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2025
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IF 3.2
Zimin Cai,Zihan Xing,Mingzhe Xu,Yuting Zhao,Lin Ye,Wei Sun,Ruijia Tao,Liuya Mi,Bowen Yang,Lei Wang,Yancui Zhao,Xiaoli Liu,Liping You
Silver nanoparticles (AgNPs) and Ag + ions are both detected in aquatic environments, posing potential risks to marine ecosystems. Mytilus galloprovincialis is an effective model for monitoring marine environments. In this study, AgNPs were synthesized using a chemical approach, and to distinguish the toxicological effects of AgNPs and AgNO 3 in M. galloprovincialis , experiments were conducted using various treatments (control, AgNO 3 , AgNPs, and AgNPs + cysteine). Our findings revealed that the uptake rate of AgNPs and AgNO 3 was different, they predominantly accumulated in the hepatopancreas and gills. qRT-PCR analysis showed varying degrees of alterations in immune genes of HSPA12A, TCTP, sHSP22, sHSP24.1 , P63, Bcl-2, and Ras . Histopathological analysis demonstrated disrupted epithelial cell arrangements and connective tissue damage in the hepatopancreas, with the AgNPs exhibiting the most severe damage compared to AgNO 3 . In addition, AgNPs significantly induced oxidative stress in hemocytes compared to AgNO 3 , resulting in elevated apoptosis rates. These findings contribute to a deeper understanding of the AgNPs and AgNO 3 interactions in marine environments and provide a theoretical basis for the evaluation of marine pollution and biomonitoring strategies.
研究领域:
海洋毒理学
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纳米毒理学
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环境科学
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分子生物学
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生态毒理学
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DOI
PubMed
高影响因子
Frontiers in Immunology
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2022
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IF 8.786
Liu Senyue,Luo Lin,Zuo Fengyuan,Geng Yi,Ou Yangping,Chen Defang,Yang Shiyong,Luo Wei,Wang Yan,Wang Jun,Huang Xiaoli
Myxobolus, a major harmful type of myxospora, is one of the main parasitic pathogens of freshwater fish. Once myxoboliosis occurs, treatment can be extremely difficult. Therefore, clear understandings of the etiology of myxoboliosis and its pathological mechanism are keys for prevention and control. Here, histology, transmission electron microscopy, transcriptome study, tunel assay, and immunohistochemistry were carried out, revealing the morphology, pathological effects as well as host response mechanism of goldfish gill to Myxobolus ampullicapsulatus. Histological studies showed that the mature spores of Myxobolus ampullicapsulatus were composed of three parts, the spore shell, sporoplasm and bottle shaped polar capsule containing double S-shaped polar filaments. Transcriptome analysis revealed that Myxobolus ampullicapsulatus -infected (Myx) goldfish gills were characterized by apoptosis activation mediated by “p53 signaling pathway” with significantly up-regulated apoptosis-related differential genes dominated by p53-Bcl2/Bax signaling pathway. In addition, tunel assay revealed severe gill apoptosis in the Myx group. Transcriptome analysis also revealed that Myx group showed changes in immune response and significantly down-regulated immune-related differential genes. Beyond that, immunohistochemistry showed that there was no significant increase in the number of gill lymphocyte after parasite infection. These results suggest that the pathological mechanism of Myxobolus ampullicapsulatus infection on gills of goldfish may be related to apoptosis and immunosuppression. Subsequent qRT-PCR showed that apoptosis-related genes (Caspase3,Bad, Bax) and anti-inflammatory gene IL-10 were significantly increased, while immune-related pro-inflammatory genes (IL-1β, IL-8) were markedly down-regulated, further verifying the transcriptome results. Based on the above results, we concluded that p53-Bcl2/Bax related networks that dominant the expression of apoptosis genes were activated while immunity was suppressed in the gills of Myxobolus ampullicapsulatus infected goldfish. Our study is not only of benefit to enrich the taxonomy of Myxobolus but also clarifies its pathogenic mechanism, thus providing targets for prevention and control of myxoboliosis.
研究领域:
鱼类病理学
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分子生物学
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寄生虫学
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免疫学
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转录组学
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DOI
PubMed
高影响因子
Evidence-based Complementary and Alternative Medicine
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2022
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IF 2.65
Qiang Shi,Jian-Gang Ju
Circular RNA (circRNA) is considered an important regulator of cancer. Circ_0001998 is a newly discovered circRNA and its role in lung adenocarcinoma (LUAD) remains obscure and requires further study. The expression levels of circ_0001998 and miR-145 in LUAD were predicted by bioinformatics analysis and then verified by qRT-PCR in the LUAD cell lines. CCK-8, clone formation, EdU assay, and flow cytometry were applied to determine the effects of silencing circ_0001998 on the viability, proliferation, and apoptosis of LUAD cells. The target relationship between circ_0001998 and miR-145 was predicted by bioinformatics analysis and verified by a luciferase activity experiment. The effect of circ_0001998/miR-145 axis on the viability, proliferation, and apoptosis of LUAD cells was verified by the rescue experiment. Circ_0001998 was upregulated in LUAD, and silencing circ_0001998 suppressed viability, proliferation, and invasion of LUAD cells. The target gene of circ_0001998, miR-145, was downregulated in LUAD, and the low expression of miR-145 indicated a poor prognosis. The effect of silencing circ_0001998 on the biological function of LUAD cells was reversed by the miR-145 inhibitor. Circ_0001998 regulates the proliferation, invasion, and apoptosis of LUAD via sponging miR-145.
研究领域:
肿瘤学
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分子生物学
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生物信息学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2024
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IF 8.5
Kehan Zhu,Zhen Yao,Tianyi Gu,Xianhao Jiang,Jianqin Zhou,Duxin Li
Swollen culms of Zizania latifolia possess a peculiar flavor, nutritional value and health-promoting effects. This study investigated the wound healing ability of polysaccharides isolated from Zizania latifolia (ZLP). Through in vitro assays, ZLP demonstrated negligible cytotoxicity and promoted the migration and phagocytosis of Staphylococcus aureus by macrophages where ZLP-2 had a significantly greater effect. Moreover, ZLP-2 has a higher purity and molecular weight, an integral and large lamellar morphology, and a larger particle size in solution, suggesting its ability to form films. Thus, ZLP-2 was fabricated as a gel formulation for topical application. The in vivo results showed that the ZLP-2 gel had significantly greater effects than did the autonomous healing process according to quantitative analysis of the wound closure area. The ZLP-2 gel decreased the levels of TNF-α and IL-6 in wounded tissue. Furthermore, the quantification of the molecular indexes via qRT-PCR revealed that the levels of tnf-α, tgf-β, mcp-1, ifn-γ, inos, and vegf in the treated group increased during the inflammatory and proliferation stages but decreased after wound closure. These results suggest that ZLP-2 has great potential in pharmaceutical applications for wound care.
研究领域:
药理学
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伤口愈合
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多糖研究
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炎症
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DOI
PubMed
高影响因子
ANNALS OF BOTANY
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2024
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IF 3.6
Guo Huanhuan,Li Shiqin,Liu Yong,Yang Qinsong
Background and AimsChinese pistachio (Pistacia chinensis Bunge), an important horticultural plant species, holds great ornamental value with beautiful leaves and fruits. Seedling propagation of this tree species is restricted by its erratic seed germination, however, the germination mechanism is ambiguous yet. The aim of this study was to figure out the germination mechanism from the novel perspective based on the multi-omics data.MethodsThe multi-omics technique combined with hormone content measurement was first applied in seed germination of Chinese pistachio.Key ResultsDue to the great accumulation during seed germination, catechin stood out from the identified metabolites by broadly targeted metabolomic analysis. Exogenous catechin of 10 mg/L significantly improved the germination of Chinese pistachio seeds. An interesting result of hormone analysis showed that the improving effect of catechin could be attributed to increase of the gibberellic acid 3 (GA3) content rather than decrease of the abscisic acid (ABA) content before germination. The paclobutrazol (PAC, a GA biosynthesis inhibitor) and PAC + catechin treatments also showed that the promoting effect of catechin on seed germination depends on GA biosynthesis. Transcriptome analysis and qRT‒PCR further revealed that catechin induced the expression of PcGA20ox5 to activate GA biosynthesis. Several transcription factors were induced by catechin and GA treatments, such as TCP, bZIP and C3H, which may play an important regulatory role in GA biosynthesis in a catechin-mediated way.ConclusionsCatechin promotes seed germination via GA biosynthesis in Chinese pistachios. This study proposes a novel mechanism by which catechin promotes seed germination via the GA pathway, which provides new insight into a comprehensive understanding of seed dormancy and germination.
研究领域:
植物学
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植物生理学
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分子生物学
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园艺学
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PubMed
高影响因子
TALANTA
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2021
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IF 6.057
Min-Xi Li,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu
Herein, a label free and sensitive miRNA detection method with enhanced practical applicability was developed based on the locked nucleic acid (LNA) assisted repeated fishing amplification strategy. The working mechanism of the proposed method is as follows: 1) a DNA probe (i.e, L-DNA) with LNA bases is immobilized onto the surface of a gold foil. The L-DNA hybridizes with the 3′ terminus of the first strands of complementary deoxyribonucleic acid (cDNA) of the target miRNA in the test samples; 2) The protruding 5′ terminus of the cDNA serves as a ‘fishhook’ to repeatedly fish the products of a hybridization chain reaction (HCR) out from a ‘reaction tube’; 3) The HCR products can be unloaded from the gold foil into a ‘product tube’ through temperature-controlled dehybridization; 4) The concentration of the target miRNA is determined based on the fluorescence intensity generated by the addition of SYBR-Green I (SG) into the ‘product tube’. The proposed platform was applied to the detection of miRNA-122 in cell lysate samples and obtained quantitative results with accuracy comparable to the quantitative reverse transcription PCR method (qRT-PCR). It is worth pointing out that the proposed platform achieved a limit of detection value of 2.9 fM for miRNA-122 by a simple but effective LNA-assisted repeated fishing amplification strategy instead of complicated enzyme-based amplification techniques. It is reasonable to expect that the proposed method provides a competitive alternative for designing practically applicable, cost-effective and label-free miRNA detection methods.
研究领域:
生物化学
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分子生物学
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纳米技术
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分析化学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）