Effects of ICMT gene silencing on the invasion and migration of human salivary adenoid cystic carcinoma cells in vitro / 口腔疾病防治

Objective @#To investigate the effect of isoprene cysteine carboxymethyltransferase (ICMT) gene on the migration and invasion of salivary adenoid cystic cancer cells (SACC) and the related mechanism, to provide experimental evidence for molecular targeted therapy of SACC.@*Methods@# Adenoid cystic cancer cells SACC-LM and SACC-83 were cultured in vitro, and siRNA was transfected into human SACC-LM and SACC-83 cells (experimental group) by transient transfection of a liposome vector. A blank control group and negative control group were set up respectively (transfected NC-siRNA). qRT-PCR was peformed to measure the mRNA expression of ICMT and RhoA in each group after transfection and to determine the silencing efficiency. The expression of ICMT, membrane RhoA, total RhoA, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and Rho associated with coiled helical binding protein kinase 1 (ROCK1) in each group was detected by Western blot. The proliferation abilityies of SACC cells was detected by CCK-8 assay. The migration and invasion ability of SACC cells were detected by comparing the relative healing area of cell scratch assay and the number of Transwell assay cells. @*Results@#After transfection of ICMT-siRNA into SACC-LM and SACC-83 cells, the expression of ICMT gene and protein in the experimental group was significantly decreased compared with the negative control group and blank control group (P<0.05), but there were no significant differences in the expression of RhoA gene and total protein among all groups (P>0.05). The expression of RhoA membrane proteins, ROCK1, MMP-2, MMP-9 in the experimental group was significantly decreased compared with that in the negative control group and blank control group (P<0.05). Cell proliferation ability was significantly decreased (P<0.05). The migration and invasion abilities were significantly decreased (P<0.05). @*Conclusion @#In vitro silencing of ICMT gene can effectively inhibit the migration and invasion of human SACC-LM and SACC-83 cells, and the mechanism may be related to RhoA-ROCK signaling pathway.

Rapid Analysis and Identification of Chemical Constituents from Siwei Tumuxiang San by HPLC-Q-Exactive-MS/MS / 中国实验方剂学杂志

Objective::To rapidly characterize and identify the components of Siwei Tumuxiang San by using high performance liquid chromatography tandem quadrupole-electrostatic field orbitrap high resolution mass spectrometry (HPLC-Q-Exactive-MS/MS). Method::Agilent ZORBAX SB-Aq column (4.6 mm×150 mm, 5 μm) was adopted and 5 mmol·L-1 ammonium acetate+ 0.1% formic acid aqueous solution-acetonitrile were used as mobile phase. According to the separation and structure identification results of chemical components based on the methods of ChemSpider and ChemicalBook database retrieval, a local database of molecular formula, molecular weight, structural formula and MS/MS spectrum information of chemical components in Siwei Tumuxiang San was established. An HPLC-Q-Exactive-MS/MS analysis method was established. The complex compounds of Siwei Tumuxiang San were identified rapidly by using Xcalibar 3.0 software, comparison of reference materials and studies on MS/MS fragment pathways. Result::A total of 110 compounds were identified from the Siwei Tumuxiang San by HPLC-Q-Exactive-MS/MS technology, including 3 sesquiterpene compounds from Inulae Radix, 16 alkaloid compunds and 38 isoprene flavonoid compounds from Sophorae Radix flavescentis, 12 triterpenoid saponin compounds, 1 catechin compounds and 4 flavonoid glycoside compounds from Rubus sachalinensis. The fragment pathways of the main types of compounds were summarized. Among them, mass spectrometry information of 31 compounds was reported for the first time. Conclusion::This study can be used to identify the target compounds and non-target compounds in Siwei Tumuxiang San systematically, accurately and quickly, which will lay a foundation for the in vivo analysis and pharmacokinetic study of the formulation.

Advances in isoprene synthase research / 生物工程学报

Isoprene emission can lead to significant consequence for atmospheric chemistry. In addition, isoprene is a chemical compound for various industrial applications. In the organisms, isoprene is produced by isoprene synthase that eliminates the pyrophosphate from the dimethylallyl diphosphate. As a key enzyme of isoprene formation, isoprene synthase plays an important role in the process of natural emission and artificial synthesis of isoprene. So far, isoprene synthase has been found in various plants. Isoprene synthases from different sources are of conservative structural and similar biochemical properties. In this review, the biochemical and structural characteristics of isoprene synthases from different sources were compared, the catalytic mechanism of isoprene synthase was discussed, and the perspective application of the enzyme in bioengineering was proposed.

Advances in metabolic engineering of Escherichia coli for isoprene biosynthesis / 生物工程学报

As an important industrial chemical, isoprene is mainly used as a precursor for synthetic rubbers. In addition, it also has wide applications in the field of pharmaceutical and chemical intermediates, food, adhesives and aviation fuel. Compared with conventional petrochemical routes, production of isoprene in microbial systems has been the research focus considering environment friendly and sustainable development features. This article summarizes the metabolic pathways and key enzymes of isoprene biosynthesis, reviews current methods and strategies in improving isoprene production of Escherichia coli, and also gives some basic ideas and expectation.

Purge-trap Gas Chromatography and Mass Spectrometric Method for Analysis of Isoprene in Natural Waters / 分析化学

Gas chromatography with mass spectrum detector ( GC-MSD) coupled with purge-and-trap system was set up to analyze the concentration of isoprene in natural waters. The best experimental conditions were established, including purge gas flow rate ( 50 mL/min ) , purge time ( 15 min ) , the optimum capillary column ( Rt-Alumina BOND/KCl) and the appropriate condition of temperature programming. When analyzing isoprene in natural waters, the precision was <4% (n=6), the detection limit was 0. 5 pmol/L and the recovery was 91%-102%. The preservative experiment showed that there was no obvious variation in sample concentrations of isoprene within 60 days. The concentrations of isoprene measured with the method ranged from 60 . 8 to 278 . 7 pmol/L in the Jiaozhou Bay and its adjacent river estuaries and from 44 . 7 to 77 . 2 pmol/L in Yellow River estuary, which was in good accord with those results reported in literatures in other coastal waters. In conclusion, the analytical method could meet the requirements of the analysis of concentration of isoprene in natural waters.

Research progress on bioreactors for plant isoprene / 中草药

Isoprenoid compounds, a kind of plant secondary metabolites, are widely distributed in nature. They play important roles not only in plant growth and development process, but also in the regulation of relationship between plants and environment. Besides its biological function, the isoprenoid compounds have great economic values in many fields, such as agriculture, medicine, chemical industry, and so on. However, the yield of many isoprenoid compounds with high ecnomic value in nature is very low, which limits the research and use of isoprenoid compounds. Now, using plant bioreactor to regulate the synthesis of isoprenoid compounds is a focus of life sciences, and there are many deep researches on synthesis and metabolic pathways of isoprenoid compounds, which could promote the isoprenoid compounds production technologies with plant bioreactor. At the same time, the development of these technologies bring more deep understanding of the isoprenoid compounds synthesis metabolic pathways. We reviewed the development of metabolic synthesis and regulation of isoprenoid compounds and research progress on bioreactor, which will provide a quick reference for isoprenoid compounds synthesis research.

Development of hot melt pressure sensitive adhesive for transdermal use and in vitro evaluation / 中国药学杂志

OBJECTIVE: To develop hot melt pressure sensitive adhesive (HMPSA) for transdermal use, and to investigate the in vitro drug release and permeation property of HMPSA based patches. METHODS: HMPSA was prepared using styrene-isoprene-styrene triblock copolymer (SIS), C5 petroleum resin hydrogenate, lanoline, liquid paraffin, dibutyl phthalate, 2, 6-ditertbutyl-cresol as material under orthogonal design. The formulation of HMPSA was screened using stickiness, melt temperature and vapor permeation rate as index. The in vitro drug release behavior and transdermal property of the optimized HMPSA were evaluated using a-asarone as model drug. RESULTS: The optimized formulation of HMPSA (HMPSA-OP) was followed as: SIS: C5 petroleum resin hydrogenate: lanoline: liquid paraffin: dibutylphthalate: 2, 6-ditertbutyl-cresol=100:140:20:40:20:2. HMPSA-OP had shown more rapid drug release than ordinary HMPSA. And the in vitro transdermal flux of HMPSA-OP was (4.75±0.84) μg · cm-2 · h-1, higher than that of ordinary HMPSA and acrylate PSA. CONCLUSION: The HMPSA-OP shows good property and was suitable to prepare transdermal patch.

Chemical constituents from marine alga Sargassum thunbergii / 中国药学杂志

OBJECTIVE: To study the chemical constituents of Sargassum thunbergii. METHODS: The compounds were isolated and purified upon extensive column chromatography using silica gel, ODS, Sephadex LH-20, and prepative HPLC, while their structures were identified on the basis of physicochemical properties and spectroscopic data analysis. RESULTS: Fifteen compounds were isolated from the EtOH extract. The structures were identified as(+)-isololiolide(1), (-)-loliolide(2), loliolide acetate(3) grasshopper ketone(4), apo-9β-fucoxanthinone(5), 3α-hydroxy-5, 6-epoxy-7-megastigmen-9-one(6), deacetylate-apo-13'-fucoxanthinone(7), apo-13'-fucoxanthinone(8),(R)-(-)-3-hydroxy-β-ionone(9) and 6 other compounds sargassum ketone(10), 2-heptenoicacid, 4,4-dihydroxy-2,3-dimethyl-7-lactone(11), (R)-2-hydroxy-3-phenylpropanamide(12), thymidine(13), fucosterol(14) and hexade-canoic acid-2β, 3β-dihydroxypropyl ester(15). CONCLUSION: Deacetylate-apo-13'-fucoxanthinone is a new compound, and compounds 3,5,6,8,9,15 are obtained from the genus Sargassum for the first time.

Preparation of testosterone hot melt pressure sensitive adhesive transdermal patch / 中国药学杂志

OBJECTIVE: To prepare testosterone hot melt pressure sensitive adhesive (HMPSA) transdermal patch and investigate its percutaneous permeability in vitro. METHODS: The matrix of thermoplastic elastomer styrene-isoprene-styrene(SIS) hot melt pressure sensitive adhesive was used for testosterone transdermal patch. The percutaneous permeability through excised nude mice skin or porcine skin in vitro was conducted by Franz diffusion cells. Cumulative permeation quantity (Q) and steady state permeation rate (Jss) were evaluated to optimize drug loading capacity and enhancer. The optimal transdermal patches were compared with reference patches with regard to percutaneous behaviors using excised nude mice skin and porcine skin. RESULTS: The optimal formulation contained 2% testosterone, 6% transdermal enhancer isopropyl myristate(IPM). Its permeation behavior in vitro followed zero-order kinetics. The permeation behavior of the optimal patches was better than the reference patches for excised nude mice skin and porcine skin. CONCLUSION: SIS HMPSA has a broad application potential for transdermal drug delivery system.

Desenvolvimento vegetativo de porta-enxertos de citros produzidos em diferentes recipientes / Vegetative development of citric rootstocks produced in different containers

O objetivo deste trabalho foi avaliar o desenvolvimento vegetativo de diferentes porta-enxertos cítricos semeados em bandejas de isopor, em comparação com a semeadura em tubetes cônicos irrigados por capilaridade. O delineamento experimental foi em blocos casualizados, e os tratamentos foram dispostos em parcelas subdivididas, testando os dois tipos de recipientes na parcela principal (tubetes e bandejas de isopor), com volume de substrato de 120cm³, e nas subparcelas três variedades de porta-enxertos cítricos ('Trifoliata' - Poncirus trifoliata [L.] Raf., citrangeiro 'FEPAGRO C37' - Poncirus trifoliata (L.) Raf. x Citrus sinensis (L.) Osbeck. e tangerineira 'Sunki' - C. sunki hort. ex Tan.). Foram utilizadas 20 plantas por subparcela e três repetições. O desenvolvimento vegetativo, 150 dias após a semeadura, foi superior, de modo geral, nas plantas cultivadas em bandejas. Ao longo do tempo, os porta-enxertos 'FEPAGRO C37' e 'Trifoliata' apresentaram crescimento em altura semelhante entre si e ambos foram superiores à tangerineira 'Sunki'. O citrangeiro 'FEPAGRO C37' foi superior aos demais porta-enxertos em relação ao diâmetro e acúmulo de massa seca.

This work aimed to compare the vegetative development of citric rootstocks sowed in isoprene trays on in stiff plastic tubes irrigated by capillarity. The experimental design was a randomized blocks with split plot scheme, testing both containers at the plot (tubes and isoprene trays) with 120cm³ of substrate volume, and, in the subplot, three citric rootstock varieties (trifoliate orange - Poncirus trifoliata [L.] Raf.), citrange FEPAGRO 'C37' - Poncirus trifoliata (L.) Raf. x Citrus sinensis (L.) Osbeck.] and tangerine 'Sunki' - C. sunki hort. ex Tan.). Twenty plant per subplot were used and three replications. After 150 days of sowing, plants cultivated in trays showed higher vegetative development. Rootstocks FEPAGRO 'C37' and trifoliate orange presented similar height growth and both were higher than tangerine 'Sunki'. Citrange FEPAGRO 'C37' showed longer diameter and dry mass accumulation in relation to other rootstocks.