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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 164-172, 2022.
Article in Chinese | WPRIM | ID: wpr-940634

ABSTRACT

ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors, and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots. MethodThe content of lobetyolin, atractylenolide Ⅲ, alcohol extract, and polysaccharides, as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis (PCA) and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots. ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest, and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen, total phosphorus, total potassium, alkali-hydrolyzable nitrogen, and available potassium (P<0.01), as well as soil organic matter, pH, available manganese, and available zinc (P<0.05). There was a positive correlation between pH and atractylenolide Ⅲ content (P<0.05). Soil total potassium was in positive correlation with alcohol extract and polysaccharide content (P<0.01). Soil available zinc was positively correlated with alcohol extract and the polysaccharide content (P<0.05). Sample sites with higher PCA scores were Pingshun, Huguan, and Longxi, which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats. ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers.

2.
Chinese Journal of Preventive Medicine ; (12): 181-186, 2020.
Article in Chinese | WPRIM | ID: wpr-787751

ABSTRACT

To analyze the epidemiological characteristics of rotavirus in children under 5 years old in China (excluding China Hong Kong, Macau and Taiwan data, the same below) from 2005 to 2018. Data on other infectious diarrhea in the country from 2005 to 2018 were downloaded from the National Notifiable Disease Report System was to build a database for report cases of rotavirus diarrhea in children under 5 years of age, and descriptive epidemiological methods were used to analyze the data. In 2005-2018, a total of 820 588 cases of rotavirus infection in children under 5 years old were reported nationwide, with male 500 944 cases, and with an average annual incidence of 63.7/100 000. The reported incidence showed a fluctuating upward trend increased from 8.4/100 000 to 178.1/100 000. The number of reporting provinces increased from 17 to 30. The reported incidence showed a peak of season from November to following February. The reported cases of rotavirus diarrhea in children under 5 months of age was 13.1%(107 845 cases), and the high-incidence age ranged from 6 months to 2 years old, accounting for 70.3% (576 874 cases), with a peak of 11-13 months (163 947 cases). The top three provinces (cities) reporting the incidence rate were Zhejiang (535.2/100 000), Guangdong (334.3/100 000) and Beijing (317.3/100 000), the provinces with the low reported case rates were Shanxi (0.9/100 000), Heilongjiang (1.6/100 000) and Liaoning (2.5/100 000), but there was no case reported in Tibet; The report cases of south region (745 526 cases) were 9.9 times north region (74 935 cases).The cases of rotavirus infection and other diarrhea pathogens were detected simultaneously accounted for 1.8% (15 030 cases) and mainly were positive for rotavirus and adenovirus (90.1%, 13 544 cases). The rate of rotavirus infection in children has increased rapidly since the age of 6 months, and 84.4% of the reported cases were infants before the age of 2 years.

3.
Journal of Experimental Hematology ; (6): 664-672, 2019.
Article in Chinese | WPRIM | ID: wpr-771901

ABSTRACT

OBJECTIVE@#To investigate the effect of Pim1 expression up-regulation on cell proliferation, apoptosis, chemotaxis and angiogenesis in acute myeloid leukemia (AML) cell line U937, and to explore the possible molecular mechanism involved, finally to estimate the Pim1 expression in primary AML cells.@*METHODS@#GFP-tagged plasmid for Pim1 overexpression and an empty vector plasmid were constructed, and then a stable Pim1 expressed U937 cell line and a control virus-infected U937 cell line were established by a lentiviral vector system. After confirming Pim1 overexpression in U937 cells, proliferation and apoptosis are determined by CCK-8 Kit and flow cytometry respectively. Transwell chemotaxis assay was used to measure the effect of Pim1 overexpression on AML cells. Flow cytometry and confocal microscopy were applied to detect the influence of Pim1 overexpression on phosphorylated CXCR4 (pCXCR4) and its location. Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression of angiogenesis and adhesion related genes in AML primary cells.@*RESULTS@#The lentivirus-infected AML cell line with Pim1 overex-pression and the control virusinfected AML cell line were established successfully. The Pim1 overexpression could enhance the proliferation and inhibit the cell apoptosis, moreover accompnied with the increasing expression of cyclin D1, phosphorylated BAD (pBAD) and pCXCR4. After SDF-1 α stimuli, Pim1 overexpression induced AML cell chemotaxis accompanied with p-CXCR4 expression and calcium influx increment. Pim1 overexpression has no effect on angiogenesis. Pim1 mRNA expression was significantly higher in AML patients than the healthy people.@*CONCLUSION@#Pim1 plays an important role in the pathogenesis of AML, which not only promotes AML cell proliferation and inhibition of apoptosis, but also enhances the chemotactic ability of leukemia cells, which closely relates with Pim1 phosphorylation of CXCR4 and the increase of intracellular calcium ion influx signals.


Subject(s)
Humans , Apoptosis , Cell Proliferation , Leukemia, Myeloid, Acute , Genetics , Proto-Oncogene Proteins c-pim-1 , Genetics , Signal Transduction , U937 Cells
4.
China Journal of Chinese Materia Medica ; (24): 4158-4164, 2016.
Article in Chinese | WPRIM | ID: wpr-272718

ABSTRACT

Using the latest 454 GS FLX platform and Titanium regent, a substantial expressed sequence tag (ESTs) dataset of Ephedra sinica was produced, and the profile of gene expression and function gene of which were investigated. A total of 48 389 reads with an average length of 373 bp were generated. These 454 reads were assembled into 18 801 unigenes, which were all 454 sequencing identified. A total number of 10 531 unigenes(56.0%) were annotated using BLAST searches (E-value≤1×10⁻⁵) against the Nr, Nt, TAIR, SwissProt and KEGG databases. With respect to genes related to ephedrine biosynthesis, 19 unigenes(encoding 9 enzymes) were found. A total of 97 putative genes encoding cytochrome P450s were also discovered. Data presented in this study will provide an important resource for the scientific community that is interested in the functional genomics and secondary metabolism of E. sinica.

5.
China Journal of Chinese Materia Medica ; (24): 4169-4174, 2016.
Article in Chinese | WPRIM | ID: wpr-272716

ABSTRACT

The open reading frame of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) was cloned from Phlegmarirus carinatus by RT-PCR method and the sequence was analyzed by bioinformatics tools. After searching the transcriptome dataset of P. carinatus, one unique sequence encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase was discovered. The primers were designed according to the cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase from the dataset. And then, the open reading frame (ORF) of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, named as PcHDR1 (GenBank Accession number:JQ957845), was cloned by RT-PCR strategy with the template of mixed RNA extracted from roots, stem and leaf of P. carinatus. The bioinformatic analysis of this gene and its corresponding protein was performed. The ORF of PcHDR1 consisted of 1 437 base pairs (bp), encoding one polypeptide with 478 amino acids. The sequence comparison showed that PcHDR1 is closest with GbHDR (Ginkgo biloba),and the sequence homology was up to 78%. Bioinformatics prediction and analysis indicated that PcHDR1 protein contained a conserved domain of LytB, without transmembrane region and signal peptides. This study cloned and analyzed 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase from P. carinatus. The result will provide a foundation for exploring the function of PcHDR1 involved in terpene biosynthesis in P. carinatus plants.

6.
Acta Pharmaceutica Sinica ; (12): 435-442, 2014.
Article in Chinese | WPRIM | ID: wpr-245065

ABSTRACT

Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.


Subject(s)
Alkaloids , Basic Helix-Loop-Helix Transcription Factors , Chemistry , Classification , Genetics , Metabolism , Flavonoids , Plants, Medicinal , Genetics , Metabolism , Terpenes , Metabolism
7.
Acta Pharmaceutica Sinica ; (12): 1520-1527, 2014.
Article in Chinese | WPRIM | ID: wpr-299103

ABSTRACT

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.


Subject(s)
Fungal Proteins , Metabolism , Fungi , Chemistry , Gene Expression Regulation, Fungal , Genes, Regulator , Protein Structure, Tertiary , Secondary Metabolism , Structure-Activity Relationship
8.
Acta Pharmaceutica Sinica ; (12): 206-210, 2013.
Article in Chinese | WPRIM | ID: wpr-235682

ABSTRACT

Cytochrome P450 (CYP450) is a key element in the Ganoderma triterpenoid biosynthetic pathway. The catalytic reaction process for CYP450 requires NADPH / NADH for electron transfer. After searching the genome dataset of Ganoderma lucidum, the unique sequence encoding CYP450 and NADPH were discovered, separately. The open reading frames of GLCYP450 and GLNADPH were cloned separately using RT-PCR strategy from G lucidum. The appropriate restriction enzyme cutting sites were introduced at the 5' and 3' ends of gene sequence. The genes of GLCYP450 and GLNADPH were recombined into the yeast expression vector pESC-URA, leading to the formation of the yeast expression plasmid pESC-GLNADPH-GLCYP450. This study provides a foundation for researching Ganoderma triterpene biosynthesis using the approach of synthetic biology.


Subject(s)
Amino Acid Sequence , Cloning, Molecular , Cytochrome P-450 Enzyme System , Genetics , DNA, Complementary , Genetics , Metabolism , Gene Expression Regulation, Fungal , Genetic Vectors , NADP , Genetics , Open Reading Frames , Plasmids , Reishi , Genetics , Metabolism , Synthetic Biology , Triterpenes , Metabolism , Yeasts , Genetics , Metabolism
9.
Acta Pharmaceutica Sinica ; (12): 211-218, 2013.
Article in Chinese | WPRIM | ID: wpr-235681

ABSTRACT

Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.


Subject(s)
Acetates , Pharmacology , Amino Acid Sequence , Cloning, Molecular , Cyclopentanes , Pharmacology , Flowers , Metabolism , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Oxylipins , Pharmacology , Panax notoginseng , Genetics , Metabolism , Phylogeny , Phytosterols , Plant Growth Regulators , Pharmacology , Plant Leaves , Metabolism , Plant Roots , Metabolism , Plant Stems , Metabolism , Plants, Medicinal , Genetics , Metabolism , Saponins , Squalene Monooxygenase , Chemistry , Genetics , Synthetic Biology , Triterpenes , Metabolism
10.
Acta Pharmaceutica Sinica ; (12): 219-227, 2013.
Article in Chinese | WPRIM | ID: wpr-235680

ABSTRACT

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.


Subject(s)
Amino Acid Sequence , Cloning, Molecular , DNA, Complementary , Genetics , Flowers , Metabolism , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Ginsenosides , Hydroxymethylglutaryl CoA Reductases , Genetics , Metabolism , Open Reading Frames , Panax , Genetics , Metabolism , Phylogeny , Plant Leaves , Metabolism , Plant Roots , Metabolism , Plant Stems , Metabolism , Plants, Medicinal , Genetics , Metabolism , Protein Structure, Secondary , Sequence Homology, Amino Acid , Synthetic Biology
11.
Acta Pharmaceutica Sinica ; (12): 1099-1106, 2013.
Article in Chinese | WPRIM | ID: wpr-259509

ABSTRACT

Research on medicinal model organism is one of the core technologies to promote the modernization of traditional Chinese medicine (TCM). The research progress of Salvia miltiorrhiza as medicinal model plant is summarized in this paper. The genome of S. miltiorrhiza is small and its life cycle is short, as well as this plant can be stably genetically transformed. Because S. miltiorrhiza possesses the important medicinal and economic values, recently the transcriptome and genome of S. miltiorrhiza have been significantly recovered. The research prospect of S. miltiorrhiza as medicinal model plant in TCM was discussed, including biosynthesis of active components and their genetic regulation, relationship between quality of TCM and ecological environments, and selective breeding of good quality lines. Furthermore, as medicinal model plant, the construction of mutant library for S. miltiorrhiza, the genome map with high quality, and the functional genome should be investigated. Accompanying modern investigation of life sciences, the platform for medicinal model plant, S. miltiorrhiza, will be promoted to be established. It is important to develop the ethnopharmacology and new drugs around the world.


Subject(s)
Alkenes , Chromosome Mapping , Abietanes , Genetics , Ethnopharmacology , Genome, Plant , Medicine, Chinese Traditional , Plants, Medicinal , Genetics , Metabolism , Polyphenols , Genetics , Salvia miltiorrhiza , Genetics , Metabolism , Transcriptome
12.
China Journal of Chinese Materia Medica ; (24): 633-639, 2013.
Article in Chinese | WPRIM | ID: wpr-350716

ABSTRACT

The authors reviewed the new technologies used for Panax genus research, including molecular identification technologies (especially for DNA barcoding), modern biotechnologies (e. g. the first generation and second generation sequencing technologies), and gene cloning and identification in this paper. These technologies have been successfully applied to species identification, transcriptome analysis, secondary metabolite biosynthetic pathway and the key enzyme function identification, indicating that the application of modern biotechnologies provide guarantee for the molecular identification of Panax genus. The application of modern biotechnologies also reveals the genetic information of transcriptome and functional genomics, and promotes the design of Panax plants genomic map. In summary, the application of the new technologies lay the foundation for clarifying the molecular mechanisms of ginsenoside biosynthesis and enforcing the in vitro synthesis of important natural products and new drugs in future.


Subject(s)
Biotechnology , Methods , Cloning, Molecular , DNA Fingerprinting , Ginsenosides , Panax , Genetics , Metabolism , Research Design
13.
Acta Pharmaceutica Sinica ; (12): 1079-1084, 2012.
Article in Chinese | WPRIM | ID: wpr-276197

ABSTRACT

Squalene synthase (SQS) is a key enzyme in plant terpenoid biosynthetic pathway. This study focused on cloning and analysis of Huperzia serrata SQS (HsSQS1) gene. After searching the transcriptome dataset of H serrata, one unique sequence encoding SQS was discovered. The primers were designed according to the transcript sequence of HsSQS1 from the H. serrata transcriptome dataset. The open reading frame of HsSQS1 was cloned using RT-PCR strategy. The bioinformatic analysis of this gene and its corresponding protein were performed. The cDNA (named as HsSQS1) contains a 1263 bp open reading frame and encodes a predicted protein of 420 amino acids. The GenBank accession number for this gene is JQ004938. HsSQS1 contains two transmembrane regions, without signal peptide. The conserved domain of squalene synthase was presented in HsSQS1. HsSQS1 was more abundant in H. serrata root than in leaf and stem. This study cloned and analyzed squalene synthase gene from H. serrata for the first time. The result will provide a foundation for exploring the mechanism ofterpenoid biosynthesis in H. serrata plants.


Subject(s)
Amino Acid Sequence , Biosynthetic Pathways , Cloning, Molecular , DNA, Complementary , Genetics , Expressed Sequence Tags , Farnesyl-Diphosphate Farnesyltransferase , Genetics , Metabolism , Genes, Plant , Genetics , Huperzia , Genetics , Molecular Sequence Data , Open Reading Frames , Phylogeny , Plant Leaves , Plant Roots , Plant Stems , Plants, Medicinal , Genetics , Triterpenes , Chemistry
14.
Acta Pharmaceutica Sinica ; (12): 1085-1091, 2012.
Article in Chinese | WPRIM | ID: wpr-276196

ABSTRACT

After searching the transcriptome dataset of Panax notoginseng, one unique sequence Pn02086 encoding UDP-glucosyltransferase (UGT), which may be involved in triterpene saponin biosynthesis, was discovered. The open reading frame of the UGT gene, named as PnUGT1, was cloned by 5'-RACE and RT-PCR method from P. notoginseng. The GenBank accession number for this gene is JX018210. The bioinformatic analysis of this gene and its corresponding protein was performed. The PnUGT1 gene contains a 1488 bp open reading frame and encodes a predicted protein of 495 amino acids. The molecular weight is 55.453 kD and the protein is unstable. In the secondary structure, the percentage of alpha helix, beta turn, random coil were 36.16%, 11.31%, 52.53%, respectively. The PnUGT1 contains 7 conserved domains predicted by InterProScan, including PSPG-box which is a unique consensus sequence of glycosyltransferases involved in plant secondary metabolism. The PnUGT1 was most likely to be located in the cytoplasm, without signal peptide and transmembrane region. Sequence alignment and phylogenetic analysis demonstrated that PnUGT1 had relative close relationship to the triterpene UDP-glucosyltransferase of Medicago truncatula (AAW56092), with the 66% similarity of conserved domain PSPG-box. PnUGT1 was more abundant in P. notoginseng leaf than in flower, stem and root. Therefore, PnUGT1 gene may be involved in notoginsenoside biosynthesis.


Subject(s)
Amino Acid Sequence , Cloning, Molecular , Computational Biology , DNA, Complementary , Genetics , DNA, Plant , Genetics , Gene Expression Regulation, Plant , Glucosyltransferases , Genetics , Metabolism , Medicago truncatula , Genetics , Metabolism , Molecular Sequence Data , Open Reading Frames , Panax notoginseng , Genetics , Phylogeny , Plant Leaves , Plants, Medicinal , Genetics , Protein Structure, Secondary , Sequence Alignment
15.
Acta Pharmaceutica Sinica ; (12): 1092-1097, 2012.
Article in Chinese | WPRIM | ID: wpr-276195

ABSTRACT

Mevalonate kinase (MVK) is a key enzyme in plant terpenoid biosynthetic pathway. This study focused on cloning and analysis of Panax notoginseng (Burk.) F. H. Chen MVK (PnMVK1) gene. After searching the transcriptome dataset of P. notoginseng, one unique sequence encoding MVK was discovered. The primers were designed according to the transcript sequence of PnMVK1 from the P. notoginseng transcriptome dataset. And then, the open reading frame of PnMVK1 was cloned from P. notoginseng by using RT-PCR strategy. The physical and chemical properties, secondary structure and three-dimensional structure of the PnMVK1 protein were forecasted and analyzed, and its structure and function were predicted. The cDNA (named as PnMVK1) contains a 1164 bp open reading frame and encodes a predicted protein of 387 amino acids. The GenBank accession number for this gene is JQ957844. No transmembrane region and signal peptide were present in PnMVK1. The conserved domain of mevalonate kinase was present in PnMVK1. PnMVK1 was more abundant in P. notoginseng root than other organisms. This study cloned and analyzed PnMVK1 gene from P. notoginseng for the first time. The result will provide a foundation for exploring the mechanism of terpenoid biosynthesis in P. notoginseng plants.


Subject(s)
Amino Acid Sequence , Cloning, Molecular , Cluster Analysis , Conserved Sequence , DNA, Complementary , Genetics , Gene Expression Regulation, Plant , Molecular Sequence Data , Molecular Weight , Open Reading Frames , Panax notoginseng , Genetics , Phosphotransferases (Alcohol Group Acceptor) , Genetics , Metabolism , Phylogeny , Plant Roots , Genetics , Plants, Medicinal , Genetics , Protein Structure, Secondary
16.
Acta Pharmaceutica Sinica ; (12): 1008-1014, 2011.
Article in Chinese | WPRIM | ID: wpr-233062

ABSTRACT

ERF family transcription factor (TF) represented ethylene-responsive protein which harbored a conserved AP2 domain. After searching the plant transcription factor database, a total of 75 unigenes was found which contained AP2 domain from the transcriptome dataset of Panax quinquefolius L. One unique sequence of ERF transcript, named as PqERF1, was cloned with entire open reading frame of 933 base pairs (bp). Protein prediction result indicated that the gene was localized in nucleus and had a conserved AP2 domain. PqERF1 gene could be induced by methyl jasmonate (MeJA) which was consistent to the inducing profile of triterpene ginsenosides. InterproScan prediction indicated that PqERF1 was probably a pathogenesis-related gene. Sequence alignment and phylogenetic analysis demonstrated PqERF1 was with high identity and had relative close relationship to the NtERF4 (Nicotiana tabacum), PhERF12 (Petunia x hybrida) and DcERF1 (Daucus carota) which was related to plant defense, regulation of secondary metabolism and the flower senescence respectively. Therefore, the gene was likely involved in regulation of secondary metabolism, plant defense and physical processes which would provide gene resource for further study on secondary metabolite synthesis and molecular breeding of P. quinquefolius.


Subject(s)
Amino Acid Sequence , Computational Biology , Daucus carota , Genetics , Metabolism , Gene Expression Regulation, Plant , Open Reading Frames , Panax , Genetics , Metabolism , Petunia , Genetics , Metabolism , Phylogeny , Plant Proteins , Genetics , Metabolism , Protein Structure, Secondary , RNA, Plant , Genetics , Sequence Alignment , Tobacco , Genetics , Metabolism , Transcription Factor AP-2 , Genetics , Metabolism
17.
Acta Pharmaceutica Sinica ; (12): 524-529, 2010.
Article in Chinese | WPRIM | ID: wpr-353363

ABSTRACT

To investigate the profile of gene expression in Salvia miltiorrhiza and elucidate its functional gene, 454 GS FLX platform and Titanium regent were used to produce a substantial expressed sequence tags (ESTs) dataset from the root of S. miltiorrhiza. A total of 46 722 ESTs with an average read length of 414 bp were generated. 454 ESTs were combined with the S. miltiorrhiza ESTs from GenBank. These ESTs were assembled into 18 235 unigenes. Of these unigenes, 454 sequencing identified 13 980 novel unigenes. 73% of these unigenes (13 308) were annotated using BLAST searches (E-value < or = 1e-5) against the SwissProt, KEGG TAIR, Nr and Nt databases. Twenty-seven unigenes (encoding 15 enzymes) were found to be involved in tanshinones biosynthesis, and 29 unigenes (encoding 11 enzymes) involved in phenolic acids biosynthesis. Seventy putative genes were found to encode cytochromes P450 and 577 putative transcription factor genes. Data presented in this study will constitute an important resource for the scientific community that is interested in the molecular genetics and functional genomics of S. miltiorrhiza.


Subject(s)
Alkenes , Cytochrome P-450 Enzyme System , Genetics , DNA, Plant , Genetics , Databases, Nucleic Acid , Abietanes , Genetics , Expressed Sequence Tags , Gene Expression Profiling , Methods , Genes, Plant , Genetics , High-Throughput Nucleotide Sequencing , Methods , Microsatellite Repeats , Genetics , Plant Roots , Genetics , Plants, Medicinal , Genetics , Polyphenols , Genetics , Salvia miltiorrhiza , Genetics , Sequence Analysis, DNA , Transcriptome , Genetics
18.
Acta Pharmaceutica Sinica ; (12): 807-812, 2010.
Article in Chinese | WPRIM | ID: wpr-354571

ABSTRACT

Herb Genome Program (HerbGP) includes a series of projects on whole genome sequencing (WGS) and post-genomics research of medicinal plants with unique secondary metabolism pathways or/and those of great medical and pharmaceutical importance. In this paper, we systematically discussed the strategy of HerbGP, from species selection, whole-genome sequencing, assembly and bioinformatics analysis, to postgenomics research. HerbGP will push study on Chinese traditional medicines into the front field of life science, by selecting a series of plants with unique secondary metabolism pathways as models and introducing "omics" methods into the research of these medicinal plants. HerbGP will provide great opportunities for China to be the leader in the basic research field of traditional Chinese medicine. HerbGP shall also have significant impacts on the R&D of natural medicines and the development of medicinal farming by analysis of secondary metabolic pathways and selection of cultivars with good agricultural traits.


Subject(s)
China , Chromosome Mapping , Genome, Plant , Genomics , Medicine, Chinese Traditional , Metabolic Networks and Pathways , Mutation , Plants, Medicinal , Genetics , Sequence Analysis, DNA
19.
Acta Academiae Medicinae Sinicae ; (6): 379-383, 2007.
Article in Chinese | WPRIM | ID: wpr-229970

ABSTRACT

<p><b>OBJECTIVE</b>To study the relationship between simian acquired immunodeficiency syndromn (SAIDS) and autoimmunity in simian immunodeficiency virus (SIV)-infected monkeys.</p><p><b>METHODS</b>Indirect immunofluorescence assays were performed to detect plasma or serum autoantibodies in SIV-infected monkeys. The heart, liver, spleen, lung, kidney, and lymph node of BALB/c mice, a strain of endothelial cell ECV304, and granulocytes were used as target antigens. These results were compared with HE stained slides of SIV-infected monkeys.</p><p><b>RESULTS</b>The levels of various autoantibodies, including anti-lymphocyte autoantibodies, anti-endothelial cell autoantibodies, and anti-granulocyte antibodies, increased after SIV infection in monkeys. Moreover, pathological examinations showed injuries in the lymphoid tissue and vascular pathological changes in cerebral cortex, submucosa of gastrointestinal tract, interstitial capillaries of myocardium, nephron of the kidney, and sinusoid cell of liver.</p><p><b>CONCLUSION</b>The increased autoantibodies and the pathological changes of tissues and organs confirm the existence of autoimmunity in SIV-infected monkeys.</p>


Subject(s)
Animals , Mice , Autoantibodies , Blood , Autoimmunity , Endothelial Cells , Allergy and Immunology , Granulocytes , Allergy and Immunology , Lymphocytes , Allergy and Immunology , Mice, Inbred BALB C , Simian Acquired Immunodeficiency Syndrome , Allergy and Immunology , Pathology , Simian Immunodeficiency Virus
20.
Chinese Journal of Hematology ; (12): 273-276, 2004.
Article in Chinese | WPRIM | ID: wpr-291450

ABSTRACT

<p><b>OBJECTIVE</b>To explore the molecular mechanisms of G(2)/M checkpoint initiated by diallyl disulfide (DADS) in HL-60 cells.</p><p><b>METHODS</b>Cell viability was determined by MTT assay. Cell cycle was assayed by flow cytometry. The expression of phospho-p38, Cdc25B and Cdc2, and p38 mRNA were measured by Western blotting and RT-PCR, respectively.</p><p><b>RESULTS</b>After treatment with DADS at 5 - 160 micro mol/L for 0 - 72 h, the growth of HL-60 cells were suppressed in a concentration-dependent manner and the inhibitory effect of DADS (20 micro mol/L) was similar to that of ATRA (10 nmol/L) (P > 0.05). Incubation of HL-60 cells with DADS (20 micro mol/L) for 12 h could activate G(2)/M checkpoint and increase the expression of phospho-p38 MAPK, followed by the expression of phospho-Cdc25B and phospho-Cdc2 (P < 0.05). SB202190, a specific inhibitor of p38 MAPK, markedly blocked the phosphorylation of p38 MAPK, Cdc25B and Cdc2 (P < 0.05).</p><p><b>CONCLUSION</b>DADS could induce the G(2)/M arrest in HL-60 cells which may be involved in the activation of p38 MAP kinase.</p>


Subject(s)
Humans , Allyl Compounds , Pharmacology , Blotting, Western , CDC2 Protein Kinase , Genetics , Metabolism , Cell Division , Cell Proliferation , Cell Survival , Disulfides , Pharmacology , Dose-Response Relationship, Drug , Enzyme Activation , Flow Cytometry , G2 Phase , Gene Expression , HL-60 Cells , Phosphorylation , Reverse Transcriptase Polymerase Chain Reaction , cdc25 Phosphatases , Genetics , Metabolism , p38 Mitogen-Activated Protein Kinases , Genetics , Metabolism
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