FOOD CONTROL
·
2025
·
IF 6.3
Kai Chen,Bin Xiao,JianZhi Luo,XueWei Yuan,Juan Zhang,Songxue Wang,Rui Zhang,Nan Wang,LiQiang Liu,AiKe Li,Ailiang Chen
Edible oil adulteration is a significant food safety concern in today's food fraud landscape. DNA-based detection methods emerging as the most reliable approach for identifying adulterated products. However, the processed oil matrix contains only trace amounts of highly degraded DNA, which significantly increases the difficulty of DNA extraction and greatly reduces the success rate of PCR amplification. To address these technical challenges, we designed species-specific primers for amplicons of varying lengths and systematically evaluated their performance in real-time PCR using DNA extracted from soybean, peanut, and rapeseed oils, followed by application in adulterated oil detection. All primer sets demonstrated high specificity, sensitivity and, most importantly, equal amplification efficiency. Results indicated that shorter amplicons were consistently detectable across all three vegetable oils, while longer amplicons were more likely to yield false negatives in real-time PCR reactions. Soybean oil showed CT values of 29 (74 bp) versus 38 (170 bp), peanut oil exhibited a CT value of 31 (54 bp) with no amplification for 86 bp, and rapeseed oil displayed a CT value of 34 (66 bp) with no signal for 101 bp. The use of shorter primers successfully facilitated the detection of adulterated oil.These findings suggest that amplicon length being a critical determinant for reliable DNA-based identification and authentication of edible oils. Thus, designing amplicons within the 50–80 bp range can significantly enhance the success rate of DNA-based identification and authentication of edible oils.
研究领域:
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
FEMS MICROBIOLOGY ECOLOGY
·
2014
·Vol. 87
·期 3
·576-585
·
IF 3.875
Liu Jing,Wang Jia-Kun,Zhu Wen,Pu Yi-Yi,Guan Le-Luo,Liu Jian-Xin
Treponema saccharophilum is a pectinolytic bacterium isolated from the bovine rumen. The abundance of this bacterium has not been well determined, reflecting the lack of a reliable and accurate detection method. To develop a rapid method for monitoring T. saccharophilum, we performed pyrosequencing of genomic DNA isolated from rumen microbiota to explore the 16S rRNA gene sequences of T. saccharophilum candidates. Species-specific primers were designed based on fifteen sequences of partial 16S rRNA genes generated through pyrosequencing with 97% or higher similarity with T. saccharophilumDSM2985 along with sequence from type strain. The relative abundance of T. saccharophilum was quantified in both in vitro and in vivo rumen systems with varied pectin-containing forages using real-time PCR. There was a clear association of T. saccharophilum with alfalfa hay, which contains more pectin than Chinese wild rye hay or corn stover. The relative abundance of T. saccharophilum was as high as 0.58% in vivo, comparable with the population density of other common rumen bacteria. It is recognized that T. saccharophilum plays an important role in pectin digestion in the rumen.
研究领域:
微生物学
·
分子生物学
·
动物科学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2018
·Vol. 19
·期 8
·2403
·
IF 3.687
Yukun Chen,Xiaoping Xu,Xiaohui Chen,Yan Chen,Zihao Zhang,Xu Xuhan,Yuling Lin,Zhongxiong Lai
Mother of FT and TFL1(MFT) belongs to phosphatidylethanolamine-binding protein (PEBP) family, which plays an important role in flowering time regulation, seed development, and germination. To gain insight into the molecular function ofDlMFTinDimocarpus longanLour., we isolatedDlMFTand its promoter sequence from longan embryogenic callus (EC). Bioinformatic analysis indicated that the promoter contained multiphytohormones and light responsive regulatory elements. Subcellular localization showed that the given the DlMFT signal localized in the nucleus, expression profiling implied thatDlMFTshowed significant upregulation during somatic embryogenesis (SE) and zygotic embryogenesis (ZE), and particular highly expressed in late or maturation stages. The accumulation ofDlMFTwas mainly detected in mature fruit and seed, while it was undetected in abortive seeds, and notably decreased during seed germination.DlMFTresponded differentially to exogenous hormones in longan EC. Auxins, salicylic acid (SA) and methyl jasmonate (MeJa) suppressed its expression, however, abscisic acid (ABA), brassinosteroids (BR) showed the opposite function. Meanwhile,DlMFTdifferentially responded to various abiotic stresses. Our study revealed thatDlMFTmight be a key regulator of longan somatic and zygotic embryo development, and in seed germination, it is involved in complex plant hormones and abiotic stress signaling pathways.Keywords:mother of FT and TFL1;longan;embryogenesis;phytohormones;stress response;quantitative real-time PCR
研究领域:
植物生物学
·
分子遗传学
·
发育生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2025
·
IF 4.5
Zibei Wu,Xufeng Zhang,Lili Wang,Jinlei Sui,Mingqi Yu,Bingzhangke Bao,Jianing Qian,Guohui Yi,Yinzheng Ma,Xiaowen He
Streptococcus mutans is one of the most significant pathogens leading to dental caries by producing highly cariogenic biofilms within the oral cavity. However, the increasing antimicrobial resistance brings great difficulties to the prevention and treatment of dental caries. Therefore, it is very important to develop new and effective antimicrobial agents from natural sources. Clausena lansium (Lour.) Skeels, a plant commonly cultivated in Southern China, is rich in various active constituents. In this study, the main constituents of the essential oils extracted from Clausena lansium leaves (EOL), seeds (EOS), peels (EOP) were precisely analyzed by gas chromatography–mass spectrometry. The antimicrobial and anti-biofilm activities of EOL, EOS, and EOP, along with their main chemical constituents, against Streptococcus mutans were comprehensively evaluated. The primary constituent in EOL was β-caryophyllene, with a content of 112.29 mg/mL. EOS was found to be abundant in sabinene and 4-terpineol, with constituents of 147.21 mg/mL and 54.36 mg/mL, respectively. In EOP, the most predominant constituent was β-phellandrene, with a content of 173.95 mg/mL. The minimum inhibitory concentrations of EOL, EOS, and EOP were 0.12, 2.00, and 1.00 mg/mL, respectively. The minimum concentrations of inhibiting at least 90% biofilm formation of EOL, EOS, and EOP was 0.12 mg/mL, 2.00, and 2.00 mg/mL, respectively. Intriguingly, the antimicrobial and anti-biofilm efficacies of the main constituents were significantly lower than those of the essential oils. By observing the dynamic changes of the biofilm formation of S. mutans treated with essential oils, it was found that the process of biofilm formation was greatly delayed. In some cases, there was even no biofilm formation. The results of real-time quantitative PCR indicated that the expression levels of brpA, gtfB, gtfC, gtfD, and luxS genes were down-regulated after treatment with the essential oils. This study reinforces the potential of Clausena lansium as a promising source for developing new antimicrobial and anti-biofilm agents. They could inhibit the proliferation and virulence of S. mutans, thereby contributing to the control of dental caries.
研究领域:
植物学
·
微生物学
·
药理学
·
分子生物学
·
牙科医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Genetics
·
2023
·
IF 3.7
Huang Hui,Ye Zhifang,Li Zhengzhao,Wang Bo,Li Ke,Zhou Kai,Cao Huiyuan,Zheng Jiaxuan,Wang Guangji
Identifying effective biomarkers in osteosarcoma (OS) is important for predicting prognosis. We investigated the prognostic value of ferroptosis-related genes (FRGs) in OS. Transcriptome and clinical data were obtained from The Cancer Genome Atlas and Gene Expression Omnibus. FRGs were obtained from the ferroptosis database. Univariate COX regression and LASSO regression screening were performed and an FRG-based prognostic model was constructed, which was validated using the Gene Expression Omnibus cohort. The predictive power of the model was assessed via a subgroup analysis. A nomogram was constructed using clinical markers with independent prognostic significance and risk score results. The CIBERSORT algorithm was used to detect the correlation between prognostic genes and 22 tumor-infiltrating lymphocytes. The expression of prognostic genes in erastin-treated OS cell lines was verified via real-time PCR. Six prognostic FRGs (ACSL5, ATF4, CBS, CDO1, SCD, and SLC3A2) were obtained and used to construct the risk prognosis model. Subjects were divided into high- and low-risk groups. Prognosis was worse in the high-risk group, and the model had satisfactory prediction performance for patients younger than 18 years, males, females, and those with non-metastatic disease. Univariate COX regression analysis showed that metastasis and risk score were independent risk factors for patients with OS. Nomogram was built on independent prognostic factors with superior predictive power and patient benefit. There was a significant correlation between prognostic genes and tumor immunity. Six prognostic genes were differentially expressed in ferroptosis inducer-treated OS cell lines. The identified prognostic genes can regulate tumor growth and progression by affecting the tumor microenvironment.
研究领域:
肿瘤学
·
基因组学
·
生物信息学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2016
·
IF 3.138
Ke Ming,Yun Chen,Fangke Yao,Jintong Shi,Jingjing Yang,Hongxu Du,Xunyi Wang,Yixuan Wang,Jiaguo Liu
To screen effective anti-duck hepatitis A virus (DHAV) drugs, we applied STMP-STPP method to prepare phosphorylated Codonopsis pilosula polysaccharide (pCPPS), the phosphorylation-modified product of Codonopsis pilosula polysaccharide (CPPS). The IR spectrum and field emission scanning electron microscope (FE-SEM) were subsequently used to analyze the structure of pCPPS. Several tests were conducted to compare the anti-DHAV activities of CPPS and pCPPS. The MTT method was used to compare the effect of the drugs on DHAV-infected duck embryonic hepatocytes (DEHs), and the Reed-Muench assay was employed to observe changes in the virulence of DHAV. We also applied real-time PCR to examine the relationship between virus replication and the expression of IFN-β. The results indicated that CPPS could not inhibit the replication of DHAV. In contrast, pCPPS increased the virus TCID 50 , inhibited viral replication and, accordingly, increased the survival rate of DEHs infected with DHAV. Because DHAV induced the expression of IFN-β, and the IFN-β expression level was positively associated with the number of DHAV, the reduction of IFN-β expression levels after pCPPS treatment demonstrated a decrease in the number of virus particles. These results indicated that pCPPS, which reduces the number of DHAV, was more effective than CPPS in anti-DHAV activity.
研究领域:
病毒学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Plant Gene
·
2021
·
IF 0
Hu Zhao,Xiaopu Shi,Cheng Shen,Caifeng Chen,Jiyuan Liu,Changqing Qu
Toon bud is rich in polyphenols and flavonoids, which not only have nutritional value but also have health benefits. However, the molecular mechanisms of flavonoid accumulation during storage remain unclear. In our report, small RNA libraries from toon buds were synthesized after three storage period (0, 3, and 5 days) at 4 °C. Overall, 303 homologous sequences with known microRNA (conserved miRNAs homology) and 19 putative novel miRNAs were obtained using Illumina sequencing technology. Furthermore, the potential target unigenes of these miRNAs were predicted. Of the predicted miRNAs and their target unigenes, 17 conserved miRNAs and their target unigenes were confirmed by quantitative real-time PCR (qRT-PCR). Exactly 11 miRNAs involved in the flavonoid pathway and 6 miRNAs targeting transcription factors displayed significant correlation with flavonoid accumulation in toon buds. The findings will be helpful for better understanding flavonoid accumulation in toon buds during storage.
研究领域:
植物科学
·
分子生物学
·
食品科学
✓ 已验证
DOI
Nanoscale
·
2019
·Vol. 11
·期 36
·16879-16885
·
IF 6.97
Han Ding,Yi An,Tao Zhao,Bing Liu,Yin Wang,Lei Zhang,Yu Wang,Yuan Zhang,Man Wang,Yanhan Dong,Longgang Hu,Bu-Chang Zhao,Peifeng Li
microRNAs are a type of evolutionarily conserved small non-coding RNA with a length of 18–25 nucleotides. In recent years, increasing studies have shown that the content of specific miRNAs in the blood changes significantly during the occurrence and development of major diseases such as cardiovascular disease and cancer. Therefore, miRNAs may serve as important new biomarkers that can be used for disease diagnosis in the future. Here, we improved the polyethylene glycol layer on the surface of a traditional silicon sphere to specifically capture miRNAs by means of a full-function microplate detector, at 100 microliters. The detection limit for specific miRNAs per liter of plasma can reach 1 fM, and simultaneous detection of 96 samples can be achieved. Compared with the traditional real-time PCR technology, our detection eliminates the complex steps of miRNA extraction, reverse transcription, amplification, etc. and avoids more human error in the detection process. Using the full-featured microwell detector, we can rapidly detect specific miRNAs in plasma, which can be used in the diagnosis of cardiovascular diseases in the future.
研究领域:
生物标志物研究
·
医学诊断
·
分子生物学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2019
·
IF 5.714
Da Lv,Yi Gu,Ming Guo,Ping Hou,Yan Li,Ronghui Wu
To better understand the toxicological activities of perfluorooctanoic acid (PFOA), we examined the effects of PFOA on apoptosis and its molecular mechanism in SMMC-7721 hepatoma cells. Cell viability was evaluated by MTT assay and apoptosis was determined by flow cytometry. Western blot and quantitative real-time PCR were used to examine the protein and gene expressions of Bax and Bcl-2. Our results showed that PFOA inhibited SMMC-7721 cell growth and induced apoptosis. PFOA treatment increased Bax expression and increased Bcl-2 expression at both gene and protein levels. Our study demonstrated that PFOA had toxic effects on SMMC-7721 cells, such as inhibiting cell proliferation and inducing apoptosis. Furthermore, we showed that PFOA-mediated induction of apoptosis involved inducing Bax and decreasing Bcl-2 expression as a molecular mechanism of its toxicological effects.
研究领域:
毒理学
·
细胞生物学
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Yuanhuan Yang,Feng Gao,Ying Liang,Lichao Guo,Yang Pan,Peng Cao,Yue Zhang
The identification of Chinese medicinal herbs occupies a crucial part in the development of the food and drug market. Although molecular identification based on real-time PCR offers good versatility and uniform digital standards compared with traditional methods, such as morphology, the dependence on large-scale equipment hinders spot detection and marketable applications. In this study, we developed a DNA nanoclaw for colorimetric detection and visible on-site identification of Chinese medicines. When specific miRNA is present, the DNAzyme is activated and cleaves the substrate strand, triggering the catalytic hairpin assembly (CHA) reaction and forming branched DNA junctions on AuNP-I. This can then capture AuNP-II through hybridization and facilitate their aggregation, resulting in a noticeable color change that is observable to the naked eye. By harnessing the dual amplification of DNAzyme and CHA, this highly sensitive nanoprobe successfully achieved specific identification of Chinese medicines. This offers a new perspective for on-site testing in the herbal market.
研究领域:
生物技术
·
纳米技术
·
药学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Practical Laboratory Medicine
·
2023
·
IF 1.9
Xu Tang,Lin-Ping Fan,Yang Liu
Introduction Group B streptococcus(GBS)often causes adverse outcomes such as urinary system infection, intrauterine infection, premature birth, and stillbirth in perinatal women. Perinatal screening of GBS is conducive to guiding clinical scientific intervention and improving delivery outcomes.This study quantitative real-time PCR (RT-qPCR) combined with magnetic separation was used for GBS detection. Materials and methods Sample pre-treatment in this study involved the utilization of magnetic separation (MS) technology, aiming to expedite the detection process and enhance detection sensitivity, and the cfb gene of group B streptococcus was used as the target gene to establish quantitative real-time PCR (RT-qPCR) to detect group B streptococcus. Results It was found that penicillin-functionalized magnetic beads had a good ability to enrich and capture group B Streptococcus.The findings revealed an exceptional detection sensitivity, with the ability to detect B streptococcus in urine samples at levels as low as 10 2 CFU/mL. Conclusions The utilization of MS technology in conjunction with the RT-qPCR (MS-RT-qPCR) assay, as demonstrated in this study, offers a viable approach for prenatal screening of group B streptococcus among perinatal women.
研究领域:
微生物学
·
分子生物学
·
妇产科学
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
Biomed Research International
·
2019
·
IF 2.197
Dongyu Xie,Ping Zhou,Lin Liu,Wenjing Jiang,Haina Xie,Liang Zhang,Donghao Xie
Suitable content of iron is essential for human body, but iron overload is associated with many kinds of diseases including chronic liver damage. Recently, researchers find that iron overload promotes hepatocyte autophagy and apoptosis. However, the mechanism of iron overload in liver damage remains unclear. In this study, Lo2 cells were selected as the research object, iron dextran was a model drug, and astragaloside IV was a therapeutic drug to explore the role of iron overload. MTT assay and Annexin/PI double staining were used to measure cell viability and apoptosis. Ultrastructure was observed by transmission electron microscopy. The expression levels of apoptosis and autophagy-related proteins were determined by real-time PCR and Western Blot. The results showed that iron dextran could significantly inhibit Lo2 cell viability and increase the apoptosis rate, while astragaloside IV could reverse the inhibition of Lo2 cell viability and decrease the apoptosis rate. Transmission electron microscopy showed a significant increase in the number of autophagosomes after administration of iron dextran, and the application of astragaloside IV reduced the production of autophagosomes. LC3II/I was significantly upregulated in the model group but decreased in the astragaloside IV treatment group, and P62 showed the opposite trend. Iron dextran significantly upregulated the expression of Bax and downregulated Bcl2, while astragaloside IV reversed this trend. Finally, the inhibition of hepcidin caused by iron dextran was counteracted by astragaloside IV. In conclusion, the experimental results show that the iron overload model mainly induces excessive autophagy and apoptosis of hepatocytes, thus causing damage to hepatocytes, but astragaloside IV plays a certain therapeutic role in reversing this damage.
研究领域:
药理学
·
肝病学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）