砂生槐种子生物碱诱导SGC-7901细胞凋亡的实验研究

目的 :研究砂生槐生物碱对SGC 790 1细胞系的增殖抑制和凋亡诱导作用。方法 :MTT比色法检测生物碱对SGC 790 1胃癌细胞增殖的抑制作用 ;荧光显微镜观察细胞的形态学改变 ;琼脂糖凝胶电泳法检测细胞凋亡过程中DNA碎片的形成 ;流式细胞仪分析并测定细胞凋亡率。结果 :砂生槐生物碱在 0 .6～ 4 .8mg·mL-1的范围内对SGC 790 1胃癌细胞的增殖有明显抑制作用 ,与SGC 790 1细胞株共培养 2 4h后 ,观察到典型的细胞凋亡形态学特征 ,DNA凝胶电泳呈现梯形条带 ,流式细胞仪检测形成一个DNA含量 <2n的分布区 ,即“亚G1峰”。结论 :砂生槐生物碱在体外明显抑制SGC 790 1胃癌细胞的增殖 ,诱导SGC 790 1细胞系凋亡的作用 ,并呈浓度。

粉防己地上部分的阿朴啡类生物碱成分

从防己科植物粉防己(Stephania tetrandra)的地上部分得到的单体,经光谱测试等结构研究手段鉴定为新化合物防己醌碱(stephadione)及己知化合物紫堇醌碱(corydione)、氧化南天竹啡碱(oxonantenine)、无根藤米里丁(cassameridine),南天竹啡碱(nantenine)和无根藤新碱(cassythicine)。该6种化合物均系首次自粉防己中得到且化学结构皆为阿朴啡生物碱类,这与该植物地下部分的化学成分迥然不同,反映了同一植物不同部位内次生代谢产物的生物合成方式有相当大的差别。

吉林乌头的化学成分研究

目的 :研究草药吉林乌头的化学成分 ,探讨其活性物质基础。方法 :采用乙醇提取、溶剂萃取、硅胶及氧化铝柱层析等方法从吉林乌头中分离化学成分 ,并通过化学和光谱法鉴定结构。结果 :从吉林乌头的根和地上部分中共分得 12个化合物 ,并用光谱方法鉴定了它们的结构。从根中分离得到的 5个化合物分别为gi gaconitine(1) ,8 acetylexcelsine(2 ) ,11 acetyl 1,19 epoxydenudatine(3 ) ,lepetine(4) ,denudatine(5 ) ,从地上部分 (含花 ,叶和茎 )分离得到的 7个化合物分别为lepenine(6) ,akirane(7) ,excelsine(8) ,β 谷甾醇 (β sitosterol,9) ,槲皮素(quercetin ,10 ) ,山柰酚 (kaempferol,11)和芦丁 (rutin ,12 )。结论 :首次从该植物中分离得到了C19二萜生物碱 (化合物 1) ,化合物 3 ,4,5 ,9,10 ,11,12均为首次从该植物中分离得到 ,化合物 2 ,4,7。

Total Synthesis of Anisodine

Aim To design a practical synthetic route of anisodine. Methods Starting from3α-hydroxy-6β-acetyltropine, anisodine was synthesized in 11 steps. Result Anisodine wasobtainded with an overall yield of 2.6 % . Conclusion Total synthetic route of anisodine wasachieved which may afford a possible route for commercial preparation of anisodine.

Cell Growth and Ajmalicine Accumulation in a Full Habituated Catharanthus roseus Cell Line C_ (20)hi

A full habituated cell line C_ 20hi was screened from 2,4_D dependent line (C_ 20D) of Catharanthus roseus (L.) G. Don. The investigation involved the cell growth, ajmalicine production and enzyme activity related to indole alkaloid biosynthesis in both cell lines. These results indicated that C_ 20hi cells grew faster than C_ 20D cells, and average ajmalicine content in C_ 20hi cells was 18.4 times more than that in C_ 20D when cultured in the production medium. In the growth medium, average ajmalicine content in C_ 20hi cells was 31.9 times more than that in C_ 20D cells, while the cell growth has no obvious difference. The comparison of enzyme activities in C_ 20hi and C_ 20D cells indicated that tryptophan decarboxylase (TDC), strictosidine synthase (SSS) and geraniol_10_dehydrogenase (G10H) activities have no close relation to ajmalicine accumulation, although the activities of these enzymes were higher when cells were cultured in the production medium than in the growth medium. The C_ 20hi cells are relatively stable in five years of culture.

Synergistic effect of oxymatrine and angiogenesis inhibitor NM-3 on modulating apoptosis in human gastric cancer cells

AIM: To investigate the synergistic effect of oxymatrine(OM) and angiogenesis inhibitor NM-3 on modulatingapoptosis in human gastric cancer cell lines SGC-7901,MKN-45, MKN-74. METHODS: Human gastric cancer lines SGC-7901,MKN-45, MKN-74 were treated with OM in the absenceand presence of NM-3. The inhibitory rates weredetected by MTT assay. Synergistic effect of OM andNM-3 on the growth of survivin, bcl-2, bax and p53 inSGC-7901 cells were examined by semiquantitative RT-PCR and Western blotting, and their growth inhibitoryeffects were also observed on SGC-7901 tumor xenograftin nude mice.RESULTS: OM combined with NM-3 exhibited asynergistic inhibitory effect on the growth of SGC-7901,MKN-45 and MKN-74 cells in a time-dependent manner.Twenty-four hours after treatment with OM, NM-3 aloneand their combination, mRNA expression of survivin andbcl-2 in SGC-7901 cells decreased, p53 mRNA expressionincreased. OM (4 g/L) combined with NM-3 significantlyincreased the expression of p53 mRNA and decreasedthe expression of survivin and bcl-2 compared witheither agent alone (193% ± 34% vs 129% ± 12%;44% ± 18% vs 92% ± 18%; 36 ± 17% vs 93% ± 23%,P < 0.05). Western blotting showed that the synergisticeffect of OM and NM-3 on protein translation of survivin,bcl-2 and p 53 was in accordance with their mRNAs.Furthermore, OM/NM-3 combination obviously exhibitedantitumor growth effect in xenografted human gastriccancer cells SGC-7901 compared with either agent alone.CONCLUSION: OM combined with NM-3 has synergisticinhibitory effects on human gastric cancer cells in vitro and can suppress the growth of xenografted human gastric cancer cells SGC-7901 in vivo.

Study on Agrobacterium tumefaciens-mediated Transformation of Brassica campestris L. with Fusion Gene Ycoil-bFGF

[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed （ Brassica campestris L. ） by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows： explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.

The progress in the chemical constituents of the genus Picrasma during 2007-2017

The plants of the genus Picrasma, comprised of nine species, are mainly distributed in tropical and subtropical regions of America and Asia. Some species of this genus are used as traditional medicine resources to cure anemopyretic cold, sore throat, dysentery, eczema, nausea, loss of appetite, diabetes mellitus, hypertension, and so on. A total of 157 chemical constituents identified from Picrasma were reviewed by Jiao WH et al. in 2007. Since then, 101 compounds were reported from the plants of the genus Picrasma. These compounds are assigned to alkaloids, quassinoids, triterpenoids, and others. This review aims to provide an updated overview on the chemical constituents of the plants of the genus Picrasma during 2007-2017.

Involvement of MAPK/ERK kinase-ERK pathway in exogenous bFGF-induced Egr-1 binding activity enhancement in anoxia-reoxygenation injured astrocytes

Objective Intravenous administration of basic fibroblast growth factor （bFGF） is effective to reduce the volume of cerebral infract due to ischemia. This study was designed to investigate the molecular mechanism, especially the signal transduction pathways, involved in this protective role of bFGF. Methods Anoxia-reoxygenation treated atrocytes were used to study the role of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase （MAPK/ERK kinase, MEK）-ERK signaling pathway after exogenous bFGF administration by Western blot. Electrophoretic mobile shift assay was used to detect the binding activity of early growth response factor-1 （Egr-1）, an important transcription factor for endogenous bFGF. Results bFGF could protect some signal transduction proteins from the oxygen-derived free radicals induced degradation. ERK1/2 was activated and involved in Egr-1 binding activity enhancement induced by exogenous bFGF. Conclusion MEK-ERK MAPK cascade may be an important signal transduction pathway contributed to bFGF induced enhancement of Egr-1 binding activity in anoxia-reoxygenation injured astrocytes.

Inhibition of tumor angiogenesis by TTF1 from extract of herbal medicine

AIM:To study the inhibition of tumor angiogenesis by 5,2,4'-trihydroxy-6,7,5'-trimethoxyflavone(TTF1) isolated from an extract of herbal medicine Sorbaria sorbifolia.METHODS:Angiogenic activity was assayed using the chick embryo chorioallantoic membrane(CAM) method.Microvessel density(MVD) was determined by staining tissue sections immunohistochemically for CD34 using the Weidner capillary counting method.The mRNA and protein levels of vascular endothelial growth factor(VEGF),vascular endothelialgrowth factor receptor 2(VEGFR2,Flk-1/KDR),basic fibroblast growth factor(bFGF),cyclo-oxygenase(COX)-2 and hypoxia-inducible factor(HIF)-1α were detected by quantitative real-time polymerase chain reaction and Western blotting analysis.RESULTS:The TTF1 inhibition rates for CAM were 30.8%,38.2% and 47.5% with treatment concentrations of 25,50 and 100 μg/embryo × 5 d,respectively.The inhibitory rates for tumor size were 43.8%,49.4% and 59.6% at TTF1 treatment concentrations of 5,10,and 20 μmol/kg,respectively.The average MVD was 14.2,11.2 and 8.5 at treatment concentrations of 5 μmol/kg,10 μmol/kg and 20 μmol/kg TTF1,respectively.The mRNA and protein levels of VEGF,KDR,bFGF,COX-2 and HIF-1α in mice treated with TTF1 were significantly decreased.CONCLUSION:TTF1 can inhibit tumor angiogenesis,and the mechanism may be associated with the down-regulation of VEGF,KDR,bFGF,HIF-1α and COX-2.

ISS-J脱硫催化剂在焦炉煤气净化系统的应用

针对焦炉煤气净化系统原采用PTV复合脱硫催化剂,存在催化剂和纯碱消耗高,脱硫净化度不稳定的问题,采用了ISS-J焦炉气专用催化剂,在不改变设备及流程的情况下,碱耗从2 550kg/d降到400kg/d,脱硫净化度保持在约5mg/m3,废液排放量5m3/d,副反应生成率由25.47%降低到2.29%。

Chemical Composition of Potato Tubers in Relation to Crop Production System and Environmental Conditions

In the years 2008-2010, a study on the quality of tubers grown in the two crop production systems, i.e., organic and integrated in two locations： Jadwisin--Central and Osiny--South-Eastern of Poland were conducted. The influence of such factors as the crop production system （organic and integrated）, the place of production （soil quality）, variety and years of research on the chemical composition, i.e., tuber dry matter, starch, vitamin C, nitrates and glycoalkaloids were tested. It has been found that the greatest impact on the chemical composition of tubers had the climatic conditions during the growing season and genotype. They affected the contents of all tested components. The place of production affected the contents of dry matter, starch, vitamin C and nitrates. This factor had no effect on the content of glycoalkaloids. The least effect on the chemical tubers composition was in crop production system. It only affected the contents of dry matter, starch and vitamin C. There was no significant effect of this factor on the nitrate and glycoalkaloids content.

Establishment and Characterization of a New Cell Line from the Kidney of Spotted Halibut Verasper variegates

A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium （MEM） supplemented with fetal bovine serum （FBS） and 10 ng ml4 basic fibroblast growth factor （bFGF）. Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number （2n=46）. The cells were successfully transfected with green fluorescent protein （GFP） reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus （LCDV） and found to be susceptible to the virus in cytopathic effect （CPE） observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.

Recent Advances in Colloidal Lubricant Detergents

Overbased lubricant detergents are important components in lubricating oil. Recently, a lot of papers about the synthesis mechanism, colloidal structure, acid neutralization and antifrictiorL properties of overbased detergents have been published with the development of experimental techniques, which can help us better understand the process of preparation and application of overbased detergents and propound new strategies for improving various performances of overbased detergents. In the future, the synthesis of environmentally friendly and multi-functional lubricant detergent using biodegradable vegetable oil instead of mineral oil as raw materials will be a primary objective for the colloidal lubricant detergent industry.This paper mainly presents the latest advances in the investigation of colloidal lubricant detergents.

Development of a Synthetic Medium for the in Vitro Culture of Bovine Embryos

The aim of this study was to develop a synthetic medium for the in vitro culture of bovine embryos, using various growth factors and cytokines （GF-CYK）： insulin-like growth factorl （IGF-Ⅰ）, insulin-like growth factorⅡ （IGF-Ⅱ）, basic fibroblast growth factor （bFGF）, leukemia inhibitory factor （LIF）, granulocyte-macrophage colony stimulating factor （GM-CSF） and transforming growth factor beta Ⅰ （TGF-β1） ＋ hyaluronan （HA） ＋ recombinant albumin （RA）. The embryos were cultured in synthetic oviduct fluid （SOF） supplemented with： treatment 1 （T1）： bovine serum albumin （BSA） ＋ insulin, transferrin and selenium （ITS） （control）; or treatment 2 （T2）： GF-CYK ＋ HA ＋ RA. The blastocyst rates were not significantly different between TI and T2, at seven days post fertilization （dpf） （28.9% ± 2.4% and 31.8% ±2.2%）, and at 8 dpf （36.5% ±2.4% and 39.1% ±1.9%）, respectively （P 〉 0.05）. The total cell number （TCN） was significantly higher with T2 than that with T1 at 7 dpf（164.9 ±5.3 and 149.7 ±4.0） and 8 dpf （182.7 ±6.4 and 165.0 ±5.5） （P 〈 0.05）. The blastocyst diameter obtained with T2 was significantly greater （P 〈 0.05） than with T1 at 7 dpf （173.3 μm ±4.9 μm and 157.2μm ±4.1 μm, respectively）, however, no significant differences were observed at 8 dpf （190.3 μm 5.2 μm and 179.7 μm ± 5.3 μm, respectively）. In conclusion, the synthetic medium （T2） shows a comparable development rate to the control medium and improves the blastocyst diameter and the TCN.

Synthesis and Biological Activity Study of New Schiff and Mannich Base Ligands Derived from Isatin and 3-Amino-1,2,4-triazole and Their Metal Complexes

Two new Mannich and Schiff base ligands namely： （1-（3＇-amino-methyl-1＇（H）-1＇,2＇,4＇-triazole）-3-（1＇（H）-l＇,2＇,4＇- triazole-3＇-yl） iminoisatin （Lt） and l-morpholinomethyl-3-（1＇（H）-1＇,2＇,4＇-triazolyl）iminoisatin （LII） were prepared from condensation reaction of the new Schiff base 3-（l＇（H）-l＇,2＇,4＇-triazole-3＇-yl）iminoisatin （SBH） （0.01 mol） with formaldehyde and 3-amino-l,2,4-triazole （3-At） or with formaldehyde and morpholine （0.01 mole each） respectively. Structures were characterized by tH NMR, CHN analyses, mass spectra and IR spectra. Metal complexes were synthesized by heating each ligand （0.02 mo with metal salts （0.01 moo in ethanol or ethanol： acetone mixture respectively. The formula of metal complexes were suggested depending on CHN and thermal analyses, metal and chloride contents, IR and UV-visible spectra, magnetic moments （laefr） and conductivity measurements. All complexes were of octahedral geometry except palladium （II） complexes which were square planar. The two ligands and some selected metal complexes showed various antibacterial and cytotoxic effects.

Toxicarioside A inhibits SGC-7901 proliferation,migration and invasion via NF-κB/bFGF signaling

AIM:To investigate the inhibitory role of toxicarioside A on the gastric cancer cell line human gastric cancer cell line(SGC-7901) and determine the underlying molecular mechanism.METHODS:After SGC-7901 cells were treated with toxicarioside A at various concentrations(0.5,1.5,4.5,9.0 μg/mL) for 24 h or 48 h,cell viability was determined by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl2H-tetrazolium bromide assay,and the motility and invasion of tumor cells were assessed by the Transwell chamber assay.Immunofluorescence staining,reverse transcription polymerase chain reaction and Western blotting were performed to detect the expression of basic fibroblast growth factor(bFGF) and fibroblast growth factor receptor-1(FGFR1),and nuclear factorkappa B(NF-κB) activation was examined by electrophoretic mobility shift assay.RESULTS:The results showed that toxicarioside A was capable of reducing cell viability,inhibiting cell growth,and suppressing cell migration and invasion activities in a time-and dose-dependent manner in SGC-7901 cells.Further analysis revealed that not only the expression of bFGF and its high-affinity receptor FGFR1 but also the NF-κB-DNA binding activity were effectively blocked by toxicarioside A in a dose-dependent manner compared with the control group(P < 0.05 or P < 0.01).Interestingly,application of the NF-κB specific inhibitor,pyrrolidinedithiocarbamate(PDTC),to SGC-7901 cells significantly potentized the toxicarioside A-induced down-regulation of bFGF compared with the control group(P < 0.05).CONCLUSION:These findings suggest that toxicarioside A has an anti-gastric cancer activity and this effect may be achieved partly through down-regulation of NF-κB and bFGF/FGFR1 signaling.

Study on the Molecular Weight Composition and Antioxidant Activity of Peanut Bioactive Peptides

[ Objective] To prepare peanut bioactive peptides and analyze their molecular weight composition and antioxidant activity. [ Method ] The dialysis bag of 8.0, 3.5 and 1.0 kD were used to classify the hydrolyzate derived from alcalase and flavourzyme, peanut bioactive poptides of differ-ent molecular weight were obtained and then their scavenging capacity of free radicals was measured. The molecular weight composition was stud-ied by Tricine-SDS-PAGE.[Result] The content of peptides 〈 1.0 kD were 76.21% and 83.42% in the total hydrolyzate from alcalase and fla-vourzyme respectively. All hydrolyzate with different molecular grades showed free radical scavenging capacity, which was increased with the reduc- tion of molecular mass. The peptides 〈 1.0 kD exhibited higher radical scavenging capacity of （87.41 ±0.66） % （alcalase） and （67.88 ±0.48）% （ flavourzyme）, respectively. [ Conclusion] Peanut bioactive peptides had strong effect of antioxidant capacity, especially that 〈 1.0 kD, which had great prospect.