Objective:Stephania tetrandra is widely used in traditional Chinese medication.In present study,we studied the resource status of it in Anhui and Jiangxi Province.Methods:We inspected relevant documents to confirm which counties we would investigate;and then,we interviewed many relative local people and carried out sample-plot survey.Results:The resource of wild Stephania tetrandra in the two provinces were both seriously damaged.Conclusion:Large-scale promotion of artificial cultivation was required to ensure the supply and to realize the reasonable and sustainable utility of this medicinal material.

OBJECTIVETo clone and sequence the open reading frame of beta-amyrin synthase (bAS) from Glycyrrhiza uralensis.

METHODThe primers were designed according to the cDNA sequence of beta-amyrin synthase from G. glabra reported by Hiroaki HAYASHI, and the open reading frame of beta-amyrin synthase was cloned by RT-PCR strategy with the template of total RNA extracted from roots of G. uralensis.

RESULTThe GubAS (GenBank Accession number: FJ627179) was 2 289 bp in length encoding one pelypeptide of 762 amino acid. Deduced amino acid sequence had 99%, 92%, 90%, 90% and 89% homology to the amino acid sequence of G. glabra, Lotus japonicus, Pisum sativum, Medicago truncatula, Glycine max, respectively.

CONCLUSIONThe open reading frame of bAS from G. uralensis is cloned and reported for the first time. The conclusion will provide a foundation for exploring the mechanism of triterpenes biosynthesis.

Cloning, Molecular ; Glycyrrhiza uralensis ; classification ; enzymology ; genetics ; Intramolecular Transferases ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants ; classification ; genetics

OBJECTIVETo analyze the correlation between content of glycyrrhizic acid and the single nucleotide polymorphism of beta-amyrin synthase (bAS) in Glycyrrhiza uralensis.

METHODglycyrrhizic acid content in 80 samples of the cultivated G. uralensis were determined by HPLC; According to the very significant level (P < 0.000 1), 80 samples in accordance with glycyrrhizic acid will be grouped by SAS 9.0; Using RT-PCR strategy to amplification the Open Reading Frame of beta-amyrin synthase with the template of total RNA extracted from roots of G. uralensis and then using DNAman to analyze the relationship between glycyrrhizic acid content and the single nucleotide polymorphism of beta-amyrin synthase (bAS).

RESULTThere exited two mutation sites 94 bp and 254 bp, G/A conversion occurred at 94 bp site, which belonged to a missense mutation. G/A conversion led to the corresponding amino acid conversion (Gly --> Asp); C/T conversion occurred at 254 bp site, which belonged to a synonymous mutation. According to sequence variation, the samples were divided into four genotypes: G-T genotype, A-T genotype, G/A-C genotype and G-T genotype.

CONCLUSIONA-T genotype, G/A-C genotype and G-T genotype are correlated with the high content of glycyrrhizic acid.

Genotype ; Glycyrrhiza uralensis ; enzymology ; genetics ; metabolism ; Glycyrrhizic Acid ; metabolism ; Intramolecular Transferases ; genetics ; Polymorphism, Single Nucleotide ; Reproducibility of Results

OBJECTIVETo analyze heterologous expression in Saccharomyces cerevisiae of two genotypes: beta-AS (A-T) genotype which is related to high content of glycyrrhizic acid and beta-AS(G-C) genotype which is related to low content of glycyrrhizic acid, and compare two different genotypes on the impact of beta-amyrin production in order to provide a foundation for licorice molecular breeding.

METHODThe 2 289 bp fragment in plasmid pMD-19T encoding beta-amyrin synthase was subcloned into the yeast-Escherichia coli shuttle vector pY26, thus an expression recombinant plasmid PY-beta-AS containing target gene was constructed. The PY-beta-AS was introduced into defective mutant INVSc1 of S. cerevisiae by LiAc method, after induced by IPTG, the content of beta-amyrin was determined by GC-MS.

RESULTGC-MS analysis demonstrates that the an occurring peak corresponding to beta-amyrin standards was detected with the same retention time, which is absent in the cell transform with empty vector. Results showed the peak was beta-amyrin and the percentage of beta-amyrin in two genotypes: beta-AS (A-T) genotype and beta-AS (G-C) genotype were 19.08% and 1.40%, respectively. Thus the beta-amyrin synthase exhibited the activity of catalyzing 2, 3-oxidosqualene to beta-amyrin.

CONCLUSIONThe catalytic efficiency of beta-AS(A-T) genotype is higher than that of beta-AS(G-C) genotype, which can lay the foundation for licorice molecular breeding.

Catalysis ; Cloning, Molecular ; Genotype ; Glycyrrhiza uralensis ; chemistry ; enzymology ; genetics ; Intramolecular Transferases ; chemistry ; genetics ; metabolism ; Plant Proteins ; chemistry ; genetics ; metabolism ; Polymorphism, Genetic ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics

OBJECTIVE: To investigate the mutation of small sequence changes in microRNA-7 gene in Chinese patients with Parkinson's disease (PD). METHODS: We analyzed miR-7 variants in 225 PD patients from Chinese Han group by DNA sequence. RESULTS: None of the patients had miR-7 variants. CONCLUSION MiR-7 variation is not associated with PD in Chinese patients.
Aged ; Base Sequence ; China ; ethnology ; Female ; Humans ; Male ; MicroRNAs ; genetics ; Middle Aged ; Molecular Sequence Data ; Mutation ; Parkinson Disease ; genetics

OBJECTIVE To explore the mechanism of steam-processed Polygonatum sibiricum polysaccharides(SPSP) in prophylaxis and treatment of mice with blood deficiency syndrome(BDS) by RNA sequencing(RNA-seq) technology. METHODS The mice were randomly divided into five groups(10 mice in each group), namely normal group, model group, SPSP groups(0.1, 0.4 g·kg−1), Danggui Buxue oral liquid(DOL) group. BDS model was induced in mice by acetylphenyl-hydrazine and cyclophosphamide. Blood routine, body weight and body temperature were tested after a consecutive administration for 14 d. The differential expressed genes(DEGs) related to anti-BDS by SPSP were screened through the transcriptome sequencing of the hepatic tissue in BDS mice. Functional annotation and enrichment analysis were performed to screen out the gene expression signaling pathways related to the treatment of SPSP on BDS mice. Quantitative polymerase chain reaction(qPCR) was used to verify the experiment. RESULTS Compared with the model group, SPSP(0.4 g·kg−1) could elevate the blood routine indexes such as red blood cell, white blood cell, hemoglobin, platelet, mean corpuscular hemoglobin concen-tration(P<0.01), and reverse the body weight and body temperature to normal(P<0.01 or P<0.05). The result of transcriptomic analysis showed that the underlying mechanism was mainly related to hematopoietic cell line, retinol metabolism, steroid hormone biosynthesis, platelet activation, B cell receptor signaling pathway, and leukocyte transendothelial migration, etc. The result of qPCR showed that SPSP(0.4 g·kg−1) could elevate the expression of JAK1, STAT1 and GATA1 mRNA (P<0.01 or P<0.05). CONCLUSION SPSP has therapeutic effects on BDS. The key DEGs in the treatment of BDS by SPSP are mainly related to the restoration of hematopoietic function, regulation of hormone and immune function. The mechanism of SPSP on treatment of BDS might be the regulation of JAK1/STAT1 signaling pathway.

Objective This study aimed to identify PAX9 variants in non-syndromic tooth agenesis families of Chi-na,as well as to analyze the genotype-phenotype of non-syndromic tooth agenesis caused by PAX9 variants,which can provide a basis for the genetic diagnosis of tooth agenesis.Methods We collected the data of 44 patients with non-syn-dromic oligodontia who underwent treatment at Stomatological Hospital of Hebei Medical University between 2018 and 2023.Whole-exome sequencing was performed on the peripheral blood of the proband and its core family members,and the variants were verified by Sanger sequencing.Pathogenicity analysis and function prediction of the variants were per-formed using bioinformatics tools.The correlation between the genotype of PAX9 variant and its corresponding pheno-type was examined by reviewing 55 publications retrieved from PubMed.The studies involved 232 tooth agenesis pa-tients with PAX9 variants.Results A novel PAX9 c.447delG(p.Pro150Argfs*62)and a reported PAX9 c.406C>T(p.Gln136*)were identified in two Chinese families.Through bioinformatics analysis and three-dimensional structural mod-eling,we postulated that the frameshift variant was pathogenic.The outcome was the premature cessation of PAX9 pro-tein,which caused severe structural and functional deficiencies.Summarizing the PAX9 genotype-phenotype relationship revealed that patients carrying the PAX9 variant commonly led to loss of the second molars.Conclusion We identified the novel PAX9 c.447delG(p.Pro150Argfs*62)in a Chinese family of non-syndromic oligodontia,expanding the known variant spectrum of PAX9.The most susceptible tooth position for PAX9 variants of tooth agenesis was the second mo-lars and the deciduous molars during the deciduous dentition.