To reveal the genetic diversity and genetic structure in Artemisia annua varieties (strains) populations, we detected the genetic polymorphism within and among eight varieties (strains) populations (192 individuals) by the approach of Start Codon Targeted Polymorphism (SCoT). The associated genetic parameters were calculated by POPGENE1.31 and the relationship was constructed based on UPGMA method. The results showed that, using 20 screened primers, a total of 145 bands were produced, of which 122 were polymorphic loci. At species level, there was a high level of genetic diversity among eight varieties (strains) populations (PPB = 84.1% ,H = 0.217 3 and H(sp) = 0.341 9). However, at the variety (strains) population level, genetic diversity was lower, the average of genetic parameters was PPB = 41.9%, H = 0.121 5, H(pop) = 0.186 8. The Nei's genetic differentiation coefficient was 0.441 0, indicate that most of the genetic variation in this species existed within the variety populations. The gene flow (N(m) = 0.633 9) was less among populations, indicating that the degree of genetic differentiation was higher. Genetic similarity coefficient were changed from 0.755 1 to 0.985 7. By clustering analysis, eight varieties (strains) were clustered into two major categories and it was also showed the same or similar genetic background varieties (strains) have a tendency to gather in the same group. Results suggest that, in variety breeding, breeders should strengthen the exchange of bred germplasm and increase mutual penetration of excellent genes, which would broaden the genetic base of A. annua.
Artemisia annua ; classification ; genetics ; Codon, Initiator ; genetics ; Genetic Markers ; genetics ; Genetic Structures ; Genetic Variation ; Genetics, Population ; methods ; Phylogeny ; Polymorphism, Genetic ; Species Specificity

To investigate the genetic diversity among wild Dipsacus asperoides in China, 66 germplasmic resources of D. asperoides were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method. The results showed that the totals of 181 bands were detected using 20 primers , among which 109 were polymorphic bands. The average percentage of polymorphic bands was 60.13%. Genetic distance changed from 0.030 6 to 0.181 4. The clustering results showed that there was no significant correlation between the classification of the wild D. asperoides and their geographical origin. The relatively high genetic diversity of D. asperoides provides the basis for breeding new varieties.
China ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Dipsacaceae ; chemistry ; classification ; genetics ; Genetic Variation ; Phylogeny ; Polymorphism, Genetic

Background People recently realized that it is important for artificial vascular biodegradable graft to bionically mimic the functions of the native vessel.In order to overcome the high risk of thrombosis and keep the patency in the clinical small-diameter vascular graft (SDVG) transplantation,a double-layer bionic scaffold,which can offer anticoagulation and mechanical strength simultaneously,was designed and fabricated via electrospinning technique.Methods Heparin-conjugated polycaprolactone (hPCL) and polyurethane (PU)-collagen type Ⅰ composite was used as the inner and outer layers,respectively.The porosity and the burst pressure of SDVG were evaluated.Its biocompatibility was demonstrated by the 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H tetrazolium bromide (MTT) test in vitro and subcutaneous implants in vivo respectively.The grafts of diameter 2.5 mm and length 4.0 cm were implanted to replace the femoral artery in Beagle dog model.Then,angiography was performed in the Beagle dogs to investigate the patency and aneurysm of grafts at 2,4,and 8 weeks post-transplantation.After angiography,the patent grafts were explanted for histological analysis.Results The double-layer bionic SDVG meet the clinical mechanical demand.Its good biocompatibility was proven by cytotoxicity experiment (the cell's relative growth rates (RGR) of PU-collagen outer layer were 102.8％,109.2％ and 103.5％,while the RGR of hPCL inner layer were 99.0％,100.0％ and 98.0％,on days 1,3,and 5,respectively) and the subdermal implants experiment in the Beagle dog.Arteriography showed that all the implanted SDVGs were patent without any aneurismal dilatation or obvious anastomotic stenosis at the 2nd,4th,and 8th week after the operation,except one SDVG that failed at the 2nd week.Histological analysis and SEM showed that the inner layer was covered by new endothelial-like cells.Conclusion The double-layer bionic SDVG is a promising candidate as a replacement of native small-diameter vascular graft.

Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA. In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy. The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein. Nine introns were found in the genomic sequence. SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones. Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase. The transcription level of SmAACT was relatively higher in root than that in stem and leaf tissues. SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs. Some special genotypes were found in Salvia miltiorrhiza from different areas. SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.
Acetyl-CoA C-Acetyltransferase ; genetics ; metabolism ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression Regulation, Plant ; Genotype ; Introns ; Oligonucleotide Array Sequence Analysis ; Open Reading Frames ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plant Stems ; enzymology ; genetics ; Plants, Medicinal ; classification ; enzymology ; genetics ; Polymorphism, Single Nucleotide ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; classification ; enzymology ; genetics

OBJECTIVETo estimate the effect of the lymph duct ligation on systemic inflammatory factors and endotoxins during intestinal ischemia-reperfusion (I/R).

METHODSMale SD rats underwent occlusion of superior mesenteric artery for 60 min followed by reperfusion for 120 min plus lymph duct ligation or not. Forty rats were randomly divided into 4 groups: group A (blank); group B (sham); group C (intestinal I/R); group D (intestinal I/R plus lymph duct ligation). Mesenteric lymph nodes were harvested for standard bacteriologic cultures. The endotoxin, D-lactate, diamine oxidase (DAO), and cytokines in serum were detected.

RESULTSThe rates of bacterial translocation to mesenteric lymph nodes were 40% in group C and 20% in group D. No positive lymph node cultures were encountered in any of group A and B. The serum cytokines (except for sICAM-1) , D-lactate, DAO and endotoxin levels were lower in group D than those in group C (P<0.05), but both were higher than those in group A and B (P<0.05).

CONCLUSIONDuring intestinal I/R injury, blockage the lymph flow from gut into bloodstream decreases the levels of cytokines, and significantly attenuates the increase in intestinal permeability.

Animals ; Disease Models, Animal ; Inflammation ; Intestinal Diseases ; metabolism ; microbiology ; pathology ; Intestines ; blood supply ; pathology ; Ligation ; Lymph Nodes ; pathology ; Lymphatic System ; surgery ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; microbiology ; pathology

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.
Animals ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA Primers ; genetics ; Drug Contamination ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo study the effects of methyl jasmonate (MJ) on the accumulation and release of tanshinones in suspension cultures of Salvia miltiorrhiza hairy roots.

METHODAfter 18 day's suspension culture of S. miltiorrhiza hairy roots induced by Agrobacterium rhizogenes ATCC15834, the chemical elicitor--methyl jasmonat was added into 6-7V suspension cultures and at the same time, tanshinones contents (including cryptotanshinone and tanshinone II(A)) on the day 2, 6 and 9, after dealing with MJ, was quantified by HPLC.

RESULTAfter dealing with MJ on the day 2, 6 and 9, the concentration of cryptotanshinone reached to 0.039, 0.204, 0.571 mg x g(-1) respectively,and tanshinone II(A) reached 0.251, 0.601 and 1.563 mg x g(-1) respectively. After 9 day's treatment by MJ, the maximum increase of cryptotanshinone and tanshinone II(A) were 23.8 fold and 6.2 fold higher than that of the control respectively.

CONCLUSIONMJ could stimulate the accumulation of tanshinones in S. miltiorrhiza and have released them into the culture medium.

Acetates ; pharmacology ; Culture Techniques ; Cyclopentanes ; pharmacology ; Diterpenes, Abietane ; Oxylipins ; pharmacology ; Phenanthrenes ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; metabolism ; Plants, Medicinal ; drug effects ; growth & development ; metabolism ; Salvia miltiorrhiza ; drug effects ; growth & development ; metabolism

OBJECTIVETo investigate the reinfection rate of Helicobacter pylori (H. pylori) in gastric mucosa by two measures of oral plaque control on patients, and to demonstrate the necessity and better method of plaque control on those patients.

METHODS148 patients suffered gastritis or gastroduodenal ulcer were assigned into test group 1 (54 patients), test group 2 (55 patients) and control group (39 patients). 13C-urea breath test proved that there were no H. pylori in their gastric mucosa. Daily plaque control was used in test group 1, oral professorial interventions were added into test group 2, neither daily plaque control nor oral professorial interventions was conducted in control group. All patients were conducted 13C-urea breath test again after half a year to determine the reinfection rate of H. pylori in gastric mucosa.

RESULTS5 patients were eliminated because of stopping mouthwash in the test group 1, 8 patients failed to control dental plaque in the test group 2. The infection rates of H. pylori in gastric mucosa of test group 1, test group 2 and control group were 67.3%, 19.1%, 82.1%, respectively. The infection rate of H. pylori of test group 2 was lower significantly than that in control group and test group 1 (chi2=33, P<0.05; chi2=31.06, P<0.05). There were no significant difference between test group 1 and control group (chi2=2.43, 0.1

CONCLUSIONDental plaque is an important source of gastric H. pylori reinfection. Dental plaque control procedures should be performed in the treatment of gastric disease correlated with H. pylori. The method of mixing professional dental plaque control and solution of mouthwash was better.

Adult ; Breath Tests ; Dental Plaque ; Gastric Mucosa ; Gastritis ; Helicobacter Infections ; Helicobacter pylori ; Humans ; Male ; Middle Aged

OBJECTIVETo study the metallothionein genes of Salvia miltiorrhiza through bioinformatics and characterization of tissue expression in regenerated shoots.

METHODMetallothionein genes were obtained by cDNA microarray analyze. BLAST was used for align, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein. Semi-quantitative RT-PCR method was used to detect the gene expression level.

RESULTTwo metallothionein genes were obtained which were contained a deduced amino acid sequence of 80 and 79 residues, named as SmMT-2a and SmMT-2b, they had a homology of 71.25%. Semiquantitative RT-PCR indicated that metallothionein genes were expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in leaf than in root and stem.

CONCLUSIONIt was the first time that metallothionein genes were obtained from S. miltiorrhiza. It provides a good basis for further functional study of S. miltiorrhiza.

Amino Acid Sequence ; Base Sequence ; DNA, Complementary ; genetics ; Gene Expression Regulation, Plant ; Genes, Plant ; Genomics ; Metallothionein ; genetics ; metabolism ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Open Reading Frames ; Plants, Medicinal ; genetics ; metabolism ; Salvia miltiorrhiza ; genetics ; metabolism ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

This paper firstly introduced the acquired full length cDNA of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase from hairy roots of Salvia miltiorrhiza (Abbr: SmCMK, GenBank number: EF534309). Results of KEGG analysis showed that SmCMK was belong to the upstream of nonemevalonate pathway, the only one kinase of the pathway. The full-length cDNA was deduced as encoding 4-(cytidine 5'-diphospho)-2-C-methylerythritol kinase (designated as SmCMK), and the sequence had a 1493 bp including 5' UTR 71 bp and 3' UTR 232 bp, an open reading frame (ORF) encoding a protein of 396 amino acid residues. The deduced protein had isoelectric point (pI) of 6.78 and a calculated molecular weight about 43 kDa, similar to cloned diterpene of CMK from other species of plants such as Mentha piperita and Lycopersicon esculentum reported previously. Real time PCR results indicated that elicitors of MJ stimulated the increase of mRNA expression of SmCMK. At the same time, results of high performance liquid chromatography (HPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy root of Salvia miltiorrhiza were increased dramatically after treated with methyl jasmonate (MJ). This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by MJ. It proved primarily that the increased expression level of mRNA of SmCMK helps to enhance tanshinones' accumulation, which will be the basis for further study on the mechanism of gene regulation of secondary metabolism of tanshinones.
Acetates ; pharmacology ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Conserved Sequence ; Cyclopentanes ; pharmacology ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; Gene Expression Regulation, Plant ; Genes, Plant ; Molecular Sequence Data ; Open Reading Frames ; Oxylipins ; pharmacology ; Phenanthrenes ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; enzymology ; Plants, Medicinal ; enzymology ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Salvia miltiorrhiza ; enzymology ; genetics ; metabolism ; Sequence Homology, Amino Acid