Treatment with traditional Chinese medicinal composite is one of the most important characteristics of traditional Chinese medicine (TCM). Studying material base of TCM composite and its mechanism is a key to modernizing the industry of Chinese medicinal herbs. The research for TCM composite can be carried out from many different angles, including multiple components, multiple actions, multiple levels and multiple targets. Such a way to study TCM composite will be beneficial to improving the theory of TCM composite, guiding clinical administration and developing new products.

OBJECTIVE:To investigate the influence of TCM Granules of the detoxification and dissipation blood stasis formula Ⅱon fulminant hepatic mitochondrial lipid peroxidation in rats with hepatic failure.METHODS:The fulminant hepatic failure rat models were established by subcutaneous injection of thioacetamide (TAA).48 Wistar rats were randomly divided into 7 groups:blank control group,model group,low dose group,medium dose group,high dose group,the lactulose treatment group,and the "Bezoar pill for resurrection" treatment group.Intragastric administration was executed 3 d before model-making,twice per day with the interval of 12 hours.They were administered for 11times.12 hours after model establishment.MDA、SOD、CAT、GSH、NO and liver necrosis area in hepatic mitochondria were determined.RESULTS:The detoxification and dissipation blood stasis formula Ⅱcould notably reduce liver necrosis area and restrain produce of hepatic mitochondrial lipid peroxidatide MDA,retrieve the activity of SOD、CAT and increase the content of GSH,NO.Moreover it shows dose-effect relation.Compared with model group,there is statistical significance(P

Objective To explore the mechanism of the Jiedu Huayu Ⅱ Decoction in actue hepatic failure.MethodsThe acute hepatic failure rat models were induced with thioacetamide (TAA).84 SPF Wistar rats were divided into 7 groups randomly:blank group,model group,low dose group,middle dose group,high dose group,lactulose group,and the Angong Niuhuang Pill group.The drugs were administrated by gavage 3 d before model-making,5.5 d in total.The expression of Bcl-2,cytochrome C in mitochondrial and cytoplasm were measured with Western Blot.ResultsThe expression of Bcl-2 reduced in mitochondrial,the expression of cytochrome C increased in cytoplasm and reduce in mitochondrial in model group compare with that of blank group.Jiedu Huayu Ⅱ Decoction was able to enhance the expression of Bcl-2 and restrain cytochrome C release from mitochondrial to cytoplasm.Moreover,it shows dose-effect relation.ConclusionThe protection of Jiedu Huayu Ⅱ Decoction for hepatocyte in acute hepatic failure rats maybe relate to the increase of Bcl-2 in mitochondrial,blocking the mitochondrial permeability transition pore to arrest cytochrome C releasing.

In order to express PebC1 in Pichia pastoris, the pebC1 sequence was amplified from genome Botrytis cinerea BC-4-2-2-1 by PCR and subcloned into the Pichua pastoris expression vector pPIC9K to generate pPIC9K-pebC1. The recombinant plasmid was linearized by Bgl II and transformed into Pichia pastoris GS115 by electroporation. Recombinant Pichia pastoris GS115/pPIC9K-pebC1 was screened by MD and G418-YPD plates and further confirmed by PCR. The protein expression was induced by methanol and analyzed by SDS-PAGE. SDS-PAGE analysis showed a special band about 39 kDa and western blotting indicated a good antigenicity of the expressed protein. Bioassay results showed that the recombinant protein PebC1 can induce resistance to gray mould disease of cucumber and Arabidopsi thaliana.
Botrytis ; chemistry ; Fungal Proteins ; biosynthesis ; genetics ; pharmacology ; Pichia ; genetics ; metabolism ; Plant Diseases ; prevention & control ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo investigate the chemical constituents of the roots of Periploca forrestii and evaluate their cytotoxicity activities.

METHODSilica gel column chromatography was employed for the isolation and purification of chemical constituents. The structures were identified on the basis of spectral data and the cytotoxicities of compounds 2-4 were investigated by several tumors cell lines including blood tumor (HL-60, CCRT-CEM), prostate tumor (PC-3, DU-145) and Melanoma (UACC-62).

RESULTFour compounds were isolated and identified as follows, lupeol-20(29)-en-3-nonadecanoate (1), peroiforoside I (2), 3beta,5beta,14beta-3OH-8beta-H-car-20(22)-enolide (3), perplocin (4).

CONCLUSIONCompound 1 is a new lupane triterpene fatty acid ester. Compounds 2-4 showed notable cytotoxicity against all tumor lines.

Cell Line, Tumor ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Molecular Structure ; Periploca ; chemistry ; Plant Roots ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

In this study, peaT1 gene was subcloned into the Pichia pastoris expression vector pPIC9K, which contained both the methanol-inducible promoter and the transcription terminator of the AOX1 gene, resulting the plasmid pPIC9K-peaT1. The recombinant plasmid was linearized by Sal I or Bgl II and transformed into P. pastoris GS115 by electroporation method. Recombinant strain was screened by Minimal Dextrose Medium and further confirmed by PCR. The gene was in frame integrated into the Pichia genome through homologous recombination, resulting the recombinant strain. Regulated by the alpha-Factor, promoter of AOX1 gene and termination signal of yeast genomic, the recombinant protein was expressed and secreted into the supernatant. The SDS-PAGE analysis indicated that the apparent molecular weight of target protein was about 35 kD. Bioassay results showed that the inhibition rate of the expressed protein against TMV was 30.37%. The supernatant was collected and then purified by anion exchange chromatography. This protein can promote seedling growth of wheat obviously.
Alternaria ; genetics ; Fungal Proteins ; biosynthesis ; genetics ; pharmacology ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Triticum ; drug effects ; growth & development

PeaT1, a protein elicitor from Alternaria tenuissima can promote plant growth and trigger systemic acquired resistance in plants. In order to expand the application of PeaT1, P43 promoter sequence and nprB signal peptide-encoding sequence were cloned from Bacillus subtilis 168 chromosomal DNA. The two sequences and peaT1 gene were spliced by overlapping extension. This product was cloned into the Escherichia coli-B. subtilis shuttle vector pHY300-PLK and the resultant recombinant expression vector (pHY43N- peaTI) plasmid was transformed into B. subtilis WB800. SDS-PAGE and Western blotting analysis showed that protein elicitor PeaT1 was expressed extracellularly in B. subtilis. This recombinant bacterial strain enhanced drought tolerance and promoted seedling growth in wheat.
Adaptation, Physiological ; drug effects ; Alternaria ; chemistry ; genetics ; Bacillus subtilis ; genetics ; metabolism ; Droughts ; Fungal Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Triticum ; growth & development