Insects
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2021
·Vol. 12
·期 10
·926
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IF 2.769
Lulu Dai,Haiming Gao,Hui Chen
Simple SummaryThe bark beetle is the most well-known pest in coniferous trees worldwide. These insects only leave the host pine bark when they disperse to locate a new host. Determining howDendroctonus armandiovercome the trees’ terpene-based defense systems has been the key problem in the study of bark beetles. Therefore, the aim of this study was to discover the molecular mechanism of insect detoxification enzymes’ ability to confer resistance to terpenes. For this purpose, the genes of cytochrome P450s, glutathione S-transferases, and carboxylesterases were studied in beetles given diets containing terpenes. The results suggest that beetles express different genes in response to terpenoids, and the responses of multiple detoxifying enzymes indicate these insects’ adaption to their chemical environment.AbstractBark beetles overcome the toxic terpenoids produced by pine trees by both detoxifying and converting them into a pheromone system. Detoxification enzymes such as cytochrome P450s, glutathione S-transferases, and carboxylesterases are involved in the ability ofDendroctonus armandito adapt to its chemical environment. Ten genes from these three major classes of detoxification enzymes were selected to study how these enzymes helpD. armandito respond to the host defenses. The expression profile of these detoxification enzyme genes was observed in adult beetles after feeding on different types of diet. Significant differences were observed between two types of seminatural diet containing the phloem of pines, and a purely artificial diet containing five monoterpenes ((−)-α-pinene, (−)-β-pinene, (+)-3-carene, (±)-limonene, and turpentine oil) also caused differential transcript levels in the detoxification enzyme genes. The results suggest that monoterpenes enter the beetles through different routes (i.e., respiratory and digestive systems) and cause the expression of different genes in response, which might be involved in pheromone metabolism. In addition, the xenobiotic metabolism in bark beetles should be considered as a system comprising multiple detoxifying enzymes.Keywords:artificial feeding;chemical defense;Chinese white pine beetle;cytochrome P450;real-time qPCR
研究领域:
昆虫学
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分子生物学
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DOI
PubMed
高影响因子
WASTE MANAGEMENT
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2021
·
IF 7.145
Junya Zhang,Tiedong Lu,Hui Zhong,Peihong Shen,Yuansong Wei
It is unadvisable to discuss the antibiotic resistance genes (ARGs) reduction in anaerobic digestion (AD) system neglecting its main purpose-methane production. The methane production improvement coupling with antibiotic resistance genes (ARGs) reduction in anaerobic digestion (AD) by zero valent iron (ZVI) were simultaneously investigated. Whether the role of ZVI on the ARGs fate was random or specific was clarified through the high-throughput qPCR (HT-qPCR). Results indicated that ZVI improved methane production and ARGs reduction by 23.9% and 25.0%, respectively. The improved methane production was associated with chemical reaction and variances of microbial community caused by ZVI, where DIET between Petrimonas , Clostridium and Syntrophomonas , Methanosarcina was established along with ACAS being enriched. ZVI specifically, not randomly, facilitated the reduction of aminoglycoside resistance genes of antibiotic inactivation and tetracycline resistance genes of ribosomal protection proteins. The specifical reduction could be attributed to enzyme activity inhibition and intracellular ionic disturbance caused by higher amounts of ZVI, although most of ARGs fate could be well explained by microbial community which contributed the most to ARGs dynamics as a whole. ZVI-based AD was a promising way for the improvement of methane production coupling ARGs reduction.
研究领域:
环境科学
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微生物学
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DOI
PubMed
高影响因子
BMC Pulmonary Medicine
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2025
·Vol. 25
·期 1
·1-11
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IF 2.8
Luo Jun,Liu Min,Ai Wen,Zheng Xiaoling,Han Lu,Huang Kuo,Zhang Changlin,Wang Peng,Fan Jinhong,Li Qianyuan,Luo Chunhua
Background Timely detection of viable Staphylococcus aureus (MSSA)/ methicillin-resistant Staphylococcus aureus (MRSA) and antimicrobial resistance are crucial for the treatment of patients with pulmonary infection. Methods In the current study, we have developed and evaluated the potential clinical applicability of the PMA TaqMan-based real-time PCR (PMA-TaqMan qPCR) assay for detection of viable MSSA/MRSA and antimicrobial susceptibility directly from bronchoalveolar lavage fluid. Results The PMA-TaqMan qPCR assay demonstrated high sensitivity with a detection limit of 800 CFU/mL within 115 min. Furthermore, the assay required only 45 min of antibiotic exposure and completed antimicrobial susceptibility testing within 160 min. Upon detection of 10 spiked bronchoalveolar lavage fluid samples treated by clindamycin after 24 h, the PMA-TaqMan qPCR assay showed better agreement with culture-based methods in detecting viable bacteria. Subsequent antimicrobial susceptibility results of bronchoalveolar lavage fluid samples spiked by 16 MRSA isolates and 11 MSSA isolates based on the current assay, compared with the standard broth microdilution method, showed an overall agreement of 100% (95% confidence intervals [CI]: 80.64 to 100) for MRSA and 90.91% (95% CI: 62.27 to 98.38) for MSSA. Conclusions Our findings demonstrate that the PMA-TaqMan qPCR assay is a highly sensitive and specific method for the direct detection of viable MSSA/MRSA and antimicrobial susceptibility testing from bronchoalveolar lavage fluid.
研究领域:
微生物学
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分子诊断
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感染性疾病
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抗菌耐药
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PubMed
高影响因子
Small Methods
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2020
·Vol. 5
·期 4
·2001047
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IF 12.13
Jianqiao Chang,Yu Zhang,Yike Li,Ziwei Han,Fei Tian,Chao Liu,Qiang Feng,Yuguang Wang,Jiashu Sun,Lu Zhang
Detection of mRNA expression in live cells during treatment is a challenging task, despite its importance in tumor biology and potential therapeutic leads. Here a multilayer ratiometric fluorescent nanomachine for live-cell perturbation and imaging of mRNA at single cell resolution is reported. The nanomachines fabricated by microfluidic approaches consist of fluorescent polymeric cores and multiple lipid layers, which can efficiently deliver siRNA and molecular beacons (MBs) to cytosol and then release the cargo in a sequential way. The siRNA molecules released from the outer lipid layers lead to silencing of multidrug resistance 1 (MDR1) gene, and the MBs from the middle lipid layers detect the presence of MDR1 mRNA. The fluorescent ratio of MBs to fluorescent polymeric cores positively correlates with the expression level of MDR1 mRNA in MCF-7/ADR cells during siRNA treatment. The nanomachines provide comparable results with traditional qPCR for quantifying mRNA, showing great potential for modulation and imaging of intratumoral mRNA in vitro and in vivo.
研究领域:
生物技术
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分子生物学
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癌症研究
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纳米技术
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PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2024
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IF 6
Zhiqing Xie,Aohuan Huang,Jihang Xie,Shubo Yu,Mengfei Chen,Jie Cai,Rong Huang,Zhenjun Zhu,Yu Ding
Bacteroides dorei is a promising candidate for the next-generation probiotics (NGP), but its accurate identification remains challenging due to its close taxonomic relationship with other Bacteroides species. This study aimed to mine the novel specific molecular target for detecting Bacteroides dorei . Using pan-genome analysis and PCR validation, species-specific target genes with a size of 193 bp were identified, and the specificity of the designed primer pair (30 bp) for this molecular target was confirmed. The PCR assay demonstrated high applicability for detecting target and/or nontarget strains in three simulated food systems, yielding no false-positive or false-negative results. Additionally, a qPCR method was developed for quantifying Bacteroides dorei , with a linear detection range from 1 × 10 8 to 1 × 10 1 CFU/mL and an R 2 value greater than 0.98. This method exhibited accurate and sensitive detection capabilities suitable for both food and feces samples. Overall, our study established PCR and qPCR assays for the rapid identification of Bacteroides dorei , advancing the exploration of Bacteroides species and the application of detection for NGPs in functional foods across various product forms.
研究领域:
微生物学
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分子生物学
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食品科学
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益生菌
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高影响因子
Frontiers in Microbiology
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2018
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IF 4.019
Hong Bin,Ba Yongbing,Niu Li,Lou Fei,Zhang Zhaohuan,Liu Haiquan,Pan Yingjie,Zhao Yong
The occurrence of antibiotic resistance genes (ARGs) as emerging contaminants is of continued concern for human health. Antibiotics used in aquaculture have promoted the evolution and spread of ARGs. This study aimed to investigate the occurrence of 37 ARGs conferring resistance to six classes of antibiotics in 94 aquatic animals from five cities in southeast coast of China. The results showed that floR, sulII, sulI, strB, strA, aadA, and tetS were identified as the prominent ARGs with the high detection frequencies ranging from 30.9 to 51.1% in total samples. Then relative expression amount of seven prominent ARGs quantified by qPCR, ranging from 0.003 to 0.065. The tetS was the most abundant ARG among the seven ARGs. Though aadA was the second highest detection frequency of ARGs, it was the lowest expression amount ARG. The occurrences and abundances of ARGs in freshwater aquatic animals were greater than those in marine, reflecting the discrepancy of cultivation pattern between the freshwater and marine aquaculture. Shanghai was considered as the most prevalent site with 16 ARGs, and Ningbo merely contained 9 ARGs without of β-lactam ARGs and quinolone ARGs, showing variations of ARGs with geographical location. Eight kinds of sulfonamides and one chloramphenicol residues were further measured in samples from Shanghai. Interestingly, no target antibiotics were found, but sulfonamides resistance genes (sulI, sulII) and chloramphenicol resistance genes (floR) persisted at aquatic animals in the absence of selection pressure. Our research firstly shows comprehensive information on the ARGs in skin microbiota of aquatic animals, which could provide useful information and a new insight for better understanding on the ARGs dissemination in aquatic animals.
研究领域:
环境科学
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微生物学
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水产养殖
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公共卫生
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DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2023
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IF 5.4
Li Zhu,Xiao-jia Li,Chao Gu,Yuan Gao,Chun-sheng Zhang,Lu-yu Wang,Nai-hong Chen,Gang Li
Ethnopharmacological relevance The main clinical manifestations of eczema include itching, erythema, swelling and pain. Currently, allergies and TH1/TH2 cytokine imbalances are significant causes of eczema. TCM believes that eczema is mainly caused by incongruity between dry and wet. Wenguanmu ointment is a classic Mongolian medicine, which mainly composed of Xanthoceras sorbifolia Bunge , Coptis chinensis Franch and Bezoar. These ingredients can clear heat and dampness, dispel wind and dehumidification, anti-inflammatoryad analgesic. In this study, it was found that Wenguanmu ointment can treat eczema with anti-inflammatory, analgesic and antipruritic. Aim of the study In this study, the content of main components in Wenguanmu ointment was tested. Moreover, the therapeutic effect and mechanism of Wenguanmu ointment on eczema model mice were studied. Materials and methods Kunming mice (25 ± 2 g) were randomly divided into 6 groups: Control group; Model group; Vehicle group; Wenguanmu ointment group; Compound dexamethasone acetate cream group; Chushizhiyang ointment group. The eczema mouse model was established by DNCB. HPLC and TLC tests were used to determine the content of the main components in Wenguanmu ointment. HE staining was used to assess skin damage in mice. In order to detect the anti-inflammatory effect of Wenguanmu ointment on eczema, The levels of IgE, TNF-α, IFN-γ, COX-2 and IL-4 in serum was measured by ELISA. Genecards and Online Mendelian Inheritance in Man databases were used to analyze potential target gene predictions, and it was speculated that Wenguanmu ointment was associated with NF−κB signaling pathway and chemokine signaling pathway. To detect this inference, RT-qPCR and western blotting were used to detect protein and mRNA levels of CKLF-1, IκB-α, and NF-κB. Results Wenguanmu ointment can repress the symptoms of eczema caused by 2, 4-dinitrochlorobenzene, and inhibit the level of serum immunoglobulin E. Simultaneously it restrain the elevation of miscellaneous pro-inflammatory cytokines and chemokines, as well as reducing the expression of CKLF-1 and NF-κB protein in the nucleus, and increasing the protein expression of IκB to improve eczema. Conclusions The ameliorating effect of Wenguanmu ointment on eczema lesions can play a importment role by inhibiting the CKLF-1/NF-κB pathway.
研究领域:
民族药理学
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药理学
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免疫学
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分子生物学
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中医药
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PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
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2020
·Vol. 43
·期 5
·831-838
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IF 2.419
Liu Bin,Wang Xufeng,Zhao Jiahao,Qin Liehao,Shi Lei,Zhou Tao,Liu Shuang,Wang Chunling
Zygosaccharomyces rouxii referred as Yeast S is an important microorganism widely applied in traditional fermented food to accelerate flavor formation. Z. rouxii 3-2 referred as Yeast S 3-2 constructed previously is a new strain having higher salt tolerance than wild type. In this study, salt stress response of synthesis key flavor metabolites 3-methylthiopropanol, 2-phenylethyl alcohol, and isoamyl acetate in Yeast S and S 3-2 were investigated based on SPME–GC–MS and RT-qPCR. Analysis of GC–MS data showed that high salinity led to increase the contents of 3-methylthiopropanol and 2-phenylethyl alcohol both in Yeast S and Yeast S 3-2, while inhibited the synthesis of isoamyl acetate, and the induced effect on Yeast S 3-2 was more pronounced than Yeast S. Furthermore, the RT-qPCR results revealed that the salinity could enhance the genes expression of ARO10 and PDC1 , and stronger effect on Yeast S 3-2 than S. However, the gene expression of AFT1 was reduced in high-salt culture conditions, which was consistent with the results of GC–MS data. The results presented in this study can provide theoretical support of Z. rouxii 3-2 application during food fermentation.
研究领域:
微生物学
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食品科学
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分子生物学
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
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IF 12.545
Hai-Yan Jia,Hong-Li Zhao,Ting Wang,Pin-Ru Chen,Bin-Cheng Yin,Bang-Ce Ye
MicroRNAs (miRNAs) play an essential role in cancer diagnosis and prognosis. Developing a new method for sensitive detection of miRNA is constantly in demand. CRISPR/Cas12a system can nonspecifically cleave single-stranded DNA after specific recognition of target DNA, showing tremendous potential in molecular diagnostics. However, CRISPR-based detection methods require synthesizing different crRNAs for detecting different targets, which limit their widespread application. Herein, we design a versatile and sensitive miRNA detection platform based on CRISPR/Cas12a system combined with a hybridization chain reaction (HCR) circuit. In this design, the HCR circuit as the signal transducer converts each miRNA into multiple DNA duplexes, which act as the activators to activate the trans -cleavage activity of Cas12a for further signal amplification . More importantly, this platform can sensitively detect different miRNAs without changing the spacer sequence of crRNA due to the fixed activators formed by HCR. In addition, the consistency between the proposed platform and RT-qPCR in miRNA detection extracted from different cell lines validated its practicability, demonstrating the potential in clinical diagnosis of cancers and monitoring therapy.
研究领域:
分子生物学
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生物技术
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癌症研究
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诊断技术
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DOI
PubMed
高影响因子
Clinical & Translational Oncology
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2021
·Vol. 23
·期 9
·1866-1873
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IF 3.405
Bai Z. M.,Li X. F.,Yang Y.,Yang Y. F.,Lv D. R.,Tang L. L.
Purpose The aim of the present study was to elucidate the functional role of hsa-miR-328-3p/STAT3 pathway in the effects of propofol on gastric cancer proliferation. Methods Bioinformatics was used to analyze the molecular expression differences of hsa-miR-328-3p/STAT3 axis in stomach adenocarcinoma ( n = 435) and normal samples ( n = 41) from TCGA database. The expression of the above molecules in gastric cancer cells SGC-7901 and normal gastric mucosal cells GES-1 was verified via qPCR. The dual-luciferase assay was carried out to confirm the interaction between hsa-miR-328-3p and STAT3. Subsequently, the cell proliferation and the expression of the above molecules in SGC-7901 and GES-1 cells were evaluated after 10 μM propofol treatment. Finally, we analyzed whether propofol still inhibited the proliferation of gastric cancer by suppressing STAT3 pathway after hsa-miR-328-3p down-regulation. Results Compared with normal samples, the expression of hsa-miR-328-3p was significantly down-regulated in stomach adenocarcinoma samples, while the expression of STAT3 and downstream target genes (MMP2, CCND1 and COX2) was up-regulated. The results were consistent with those in GES-1 and SGC-7901 cell lines. Meanwhile, we found that hsa-miR-328-3p can bind to the 3′-UTR of the potential target gene STAT3. Furthermore, propofol significantly inhibited the proliferation of gastric cancer cell line SGC-7901, where hsa-miR-328-3p was up-regulated and the expression of STAT3 and downstream proliferation-related target genes were down-regulated. However, the growth inhibition of propofol on SGC-7901 cell was significantly reversed after the inhibition of hsa-miR-328-3p. Conclusions To sum up, propofol suppressed the STAT3 pathway via up-regulating hsa-miR-328-3p to inhibit gastric cancer proliferation.
研究领域:
肿瘤学
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分子生物学
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药理学
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PubMed
高影响因子
DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE
·
2025
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IF 1.8
Taobo Liang,Junyu Cao,Liyan Zhuo,Chunhong Zhu,Yufei Wang,Shanshan Hu,Jiaxing Hao,Guangsheng Zhuo,Tao Wang
Microbial contamination during the collection of placental and umbilical cord tissues presents a significant challenge to the safety of stem cell-based therapies and the commercial storage. In this study, microbial contamination analysis of a large cohort placental and umbilical cord samples collected recent revealed with 38 distinct microbial species identified. Contaminated samples were predominantly from vaginal deliveries (82.7%), and higher contamination rates were observed in mothers aged ≥30 years. For rapid monitoring, we developed a novel multiplex quantitative PCR assay for simultaneous detection of the most prevalent contaminants: Escherichia coli, Candida albicans , and Enterococcus faecalis . The assay demonstrated robust detection limits in pure cultures, with a lower limit of detection of 10 2 CFU/mL for E. coli and C. albicans , and 10 1 CFU/mL for E. faecalis . Based on the analytical performance described above, this multiplex qPCR assay establishes a robust framework for rapid, quantitative surveillance of microbial contaminants in placental-derived biologics. By enabling same-shift screening with sensitivity down to 10 CFU/mL, the method significantly enhances quality assurance protocols for stem cell banking a critical advancement toward safer clinical translation of placental-based therapeutics.
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高影响因子
Frontiers in Microbiology
·
2019
·
IF 4.259
He Shengnan,An Tai,A Runa,Liu Shouan
The tea plant [Camellia sinensis (L.) O. Kuntze] is one of the most important leaf crops, and it is widely used for the production of non-alcoholic beverages worldwide. Tea also has a long history of medicinal use. Colletotrichum camelliae Massee is one of the dominant fungal pathogens that infects tea leaves and causes severe tea anthracnose disease. To analyze the molecular biology of C. camelliae, the quantification of pathogen gene expression by the RT-qPCR method is necessary. Reliable RT-qPCR results require the use of stable reference genes for data normalization. However, suitable reference genes have not been reported in C. camelliae thus far. In this study, 12 candidate genes (i.e., CcSPAC6B12.04c, CcWDR83, Cchp11, Ccnew1, CcHplo, CcRNF5, CcHpcob, CcfaeB-2, CcYER010C, CcRNM1, CcUP18, and CcACT) were isolated from C. camelliae and assessed as potential reference genes. The expression stability of these genes in C. camelliae during spore germination and mycelial growth and interaction with host plants was first evaluated using several statistical algorithms, such as geNorm, NormFinder, and Bestkeeper. A web-based analysis program, Refinder, was then used to find the most suitable reference genes. Our results indicated that Cenew1, CcHplo, and CcSPAC6B12.04c were the most stable reference genes in C. camelliae under all conditions. Our work provided the most suitable reference genes for future studies performed to quantify the target gene expression levels of C. camelliae.
研究领域:
植物病理学
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分子生物学
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DOI
PubMed
高影响因子
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