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@#Objective:To explore the mechanism by which SRY-related high mobility group-box 9 (SOX9) promotes the epithelial mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) A549 cells via the Wnt/β-catenin pathway. Methods: The human NSCLCA549 cell line was divided into three groups: OE-NC group, OE-SOX9 group and OE-SOX9+XAV-939 group. The cells in OESOX9 group were transfected with SOX9 pcDNA plasmid to up-regulate the expression level of SOX9; The cells in OE-SOX9+XAV939 group were transfected with SOX9 pcDNA plasmid while the β-catenin inhibitor XAV-939 (1.0 μmol/L) was added to the medium. qPCR was used to detect SOX9 mRNA levels; CCK-8 was used to examine the proliferation of A549 cells; Wound-healing assay and Transwell chamber assay were used to detect the migration and invasion ofA549 cells, respectively; and WB was used to detect protein expressions of SOX9, β-catenin, E-cadherin, γ-catenin, N-cadherin and vimentin. Results: The mRNA and protein levels of SOX9 in OE-SOX9 group and OE-SOX9+XAV-939 group were significantly higher than those in the OE-NC group after transfection (all P< 0.05), while there was no significant difference between the OE-SOX9 group and the OE-SOX9+XAV-939 group (P>0.05). The proliferation, migration and invasion of cells in OE-SOX9 group were significantly higher than those in OE-NC group; however, those abilities in OE-SOX9+XAV-939 group were significantly lower than those in OE-SOX9 group (all P<0.05). The level of β-catenin protein in OE-SOX9 group was significantly higher than that in the OE-NC group, while the level of β-catenin protein in OE-SOX9+XAV-939 group was lower than that in OE-SOX9 group (all P<0.05). Compared with the OE-NC group, the levels of phenotypic markers of epithelial cells, E-cadherin and γ-catenin, were down-regulated, and the phenotypic markers of mesenchymal cells, N-cadherin and vimentin, were up-regulated in cells of OE-SOX9 group; however, E-cadherin and γ-catenin were higher, and N-cadherin and vimentin were lower in OE-SOX9+XAV-939 group than those in OE-SOX9 group (all P<0.05). Conclusion: SOX9 could promote proliferation, migration and EMT of NSCLCA549 cells by activating the Wnt/β-catenin pathway. ··
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Objective To understand microcystins (MCs) pollution in important waters of Shaoxing, so as to provide evidence for risk assessment and supervision of water quality safety. Methods Water samples were collected from 15 points set in three Shaoxing waters during September 2016 to September 2017. Six kinds of MCs (MC-LR, MC-RR, MC-YR, MC-LA, MC-LY, MC-LF) in samples were detected by ultra performance liquid chromatography tandem mass spectrometry, and the results were compared according to different time and place. Results MCs were detected positive in 22 of the 135 samples , the total detection rate was 16.3％.The detection rates of Xiaoshao River, Puyang River and Cao′e River were 7.9％, 18.5％和26.7％ respectively; the detection rates in July, August and September were 13.3％, 40％ and 46.7％ respectively, and the other months were not detected. In 22 positive samples, the detection rates of 6 MCs from high to low as follows: MC-LR (100％), MC-RR (100％), MC-YR (22.7％), MC-LY (18.2％), MC-LF (13.6％) and MC-LA (0％) . Conclusion In the three important waters of Shaoxing, microcystins were polluted in summer and autumn, which were mainly MC-LR and MC-RR.
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Persicae semen has been used for years as a traditional Chinese medicine to treat diseases. Because of their similar morphologies, Persicae semen was commonly inadvertently mixed with Armeniacae semen amarum (a toxic herbal seed). Development of a reliable method for discriminating Persicae semen from its adulterant is necessary to reduce confusion for the drug safety in clinical practices. This study evaluates the efficiency of high-resolution melting (HRM) combined with internal transcribed spacer 2 (ITS2) to analyze Persicae semen. Our findings show that HRM allows not only the identification of adulteration but also the quantification of the most common admixture. HRM sensitivity in adulterant detection was assessed through the analysis of mixing samples with different proportions of Prunus persica and Prunus armeniaca control. The results are presented as a linear regression with r of 0.96 and imply the capability of the method to detect adulteration. In particular, HRM detected seeds of Prunus persica in Prunus armeniaca at concentrations as low as 1%, and commercial products labeled as ‘Persicae semen’ were purchased from markets and could rapid authenticated by HRM analyses. This study is significant in the verification of the authenticity in the quality control of herbal medicine. In the near future, it is promising to be the main trend for identifying traditional Chinese medicinal materials.
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Hyoscyami Semen, the mature dried seed of Hyoscyamus niger L., has long been used as a traditional Chinese medicine to treat human diseases. Hyoscyami Semen is found in local markets in China. In markets, sellers and buyers commonly inadvertently mix the seeds of H. niger with the seeds of related species such as Hygrophila salicifolia (Vahl) Nees, Astragalus complanatus R. Br., Cuscuta australis R. Br., Cuscuta chinensis Lam., and Impatiens balsamina L. because of their similar morphologies or similar names. Thus, developing a reliable method for discriminating H. niger seeds from its adulterants is necessary to reduce confusion and ensure the safe use of Hyoscyami Semen. The present study was designed to evaluate the efficiency of high-resolution melting analysis combined with DNA barcoding (Bar-HRM) with internal transcribed spacer 2 to discriminate H. niger. Our results show that Bar-HRM successfully identified the adulterants and detected the proportion of H. niger DNA extract within an admixture. In particular, HRM detected H. niger DNA extract in A. complanatus DNA extract at concentrations as low as 1%. In conclusion, the Bar-HRM method developed in the present study for authenticating H. niger is rapid and cost-effective. It can be used in the future to guarantee the purity of Hyoscyami Semen for the clinical use.
Subject(s)
China , DNA Barcoding, Taxonomic , Methods , DNA, Intergenic , Chemistry , Genetics , DNA, Plant , Chemistry , Genetics , Discriminant Analysis , Drug Contamination , Drugs, Chinese Herbal , Chemistry , Hyoscyamus , Genetics , Seeds , Genetics , Transition TemperatureABSTRACT
High-resolution-melting analysis (HRM) is a new technology derived from qPCR and is widely used in the study of polymorphism, genotyping, and single nucleotide mutation. Advantages of HRM include cost-effectiveness and time-efficiency over PCR-based genotyping. However, the application of HRM in the authentication of herbal products is still limited with few studies on the classification and identification of herbal products. In this study, Cimicifugae Rhizoma was used as an example to verify the stability and accuracy of HRM technique in identification of Chinese materia medica. HRM assay was established for identification based on ITS2 region of Cimicifugae Rhizomas and its adulterants (including 41 samples). Our findings showed that HRM allows not only the identification of adulteration but also the quantification of the most common admixture. This study is significant for better quality in the verification of the authenticity of herbal medicine. The method is promising for future identification of traditional Chinese medicinal materials.
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Objective To summarize the endoscopic characteristics of Primary Gastrointestinal Malignant Lymphoma (PGIML) and increase the diagnostic accuracy of this method to aid the clinical treatment.Methods The clinical data of 67cases with PGIML from 2004 to 2009 were studied retrospectively.ResultsPGIML lacked specific clinical symptoms.The main clinical symptoms were abdominal pain, melena and abdominal mass.In our study, most PGIML(37/67,55.8％)were detected in stomach.Nodular appearance of mucosal surface was the most common endoscopic finding.The positive rate of endoscopic biopsy for the diagnosis of PGIML was 68.7 ％ (46/67 biopsy cases).All cases had Non-Hogkin Lymphoma (NHL).The most common immunophenotype was B-cell majority (50/67).Thirty cases had MALT.ConclusionIt is hard to diagnose PGIML at early stage,combination of various diagnosis tests should be considered to solve this problem.The effectiveness of surgery on PGIML should be further confirmed by prospective randomized clinical trials.