1.Molecular identification of heparin from pigs by allele-specific PCR (AS-PCR) and amplification refractory mutation system (ARMS).
Zhi-Yong YU ; Ge DING ; Xiao-Yu DING ; Bi-Hai CHU ; Liang QIAN ; Sun GU
Acta Pharmaceutica Sinica 2008;43(5):535-541
The aim of this study is to develop a convenient and effective method for the identification of heparin from pigs (include Sus scrofa domestica Brisson and Sus scrofa riukiuanus). Based on sequences of D-loop region of pigs and the other animals, two pairs of highly specific primers were designed for distinguishing heparin of pigs from other animals. The primers were employed to amplify D-loop region of DNA templates extracted from pig and seven other animal species that amounted to 49 samples. AS-PCR (allele-specific PCR) and ARMS (amplification refractory mutation system) were all suitable for fast identification of heparin from pig with anneal temperature at 54-56 degrees C in AS-PCR and with wider anneal temperature in ARMS,at 52-58 degrees C. An about 170 bp DNA fragments were amplified from separately pigs and whereas no DNA fragment was amplified from other samples under the same reaction condition.
Alleles
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Animals
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Base Sequence
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DNA Mutational Analysis
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methods
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DNA Primers
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DNA, Mitochondrial
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genetics
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Drug Contamination
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prevention & control
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Heparin
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analysis
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genetics
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Horses
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genetics
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction
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methods
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Quality Control
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Ruminants
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genetics
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Sequence Alignment
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Sequence Analysis, DNA
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Sus scrofa
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genetics
2.Genetic diversity and molecular authentication of wild populations of Dendrobium officinale by RAPD.
Ge DING ; Xiao-yu DING ; Jie SHEN ; Feng TANG ; Dong-yang LIU ; Jia HE ; Xue-xia LI ; Bi-hai CHU
Acta Pharmaceutica Sinica 2005;40(11):1028-1032
AIMGenetic diversity, relationship and molecular authentication of total 8 wild populations of Dendrobium officinale were investigated using RAPD markers.
METHODS10 random decamer primers were screened for Random Amplified Polymophic DNA (RAPD) fragments. A DNA molecular dendrogram was established based on cluster analysis by UPGMA (unweighted pair-group method with arithmetic average), and the relationship of the wild populations were analyzed, and all the wild populations were authenticated.
RESULTSA total of 439 loci with an average of 43.9 loci per primer and 54.9 loci per population were amplified from 8 wild populations by 10 effective primers. In the total 104 amplified bands, 95 were polymorphic, corresponding to 91.35% genetic polymorphism. The genetic distances were 0. 590 to 0. 727, with an average of 0. 686.
CONCLUSIONDistinct genetic differences and extensive genetic diversity were presented among the wild populations. RAPD markers were an informative and useful tool for the genetic diversity, evaluation and authentication of wild populations of Dendrobium officinale. Primer S412 could be used to authenticate 8 wild populations completely.
China ; Cluster Analysis ; DNA Fingerprinting ; DNA Primers ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Ecosystem ; Genetic Markers ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; Random Amplified Polymorphic DNA Technique
3.Impact of fenofibrate on NO and endothelial VCAM-1 expression in hyperlipidemic rats.
Jun WU ; Ming SUN ; Jin-chao LIN ; Zhao-chu HE ; Bi-ru OU ; Hai-sen GUO
Journal of Southern Medical University 2007;27(12):1872-1874
OBJECTIVETo investigate the mechanism of hyperlipidemia- and imflammation-induced functional impairment of the endothelium.
METHODSThe experiment was conducted using 3 groups of rats fed for 20 weeks with standard chow (control group), high-fat diet and high-fat diet with daily fenofibrate treatment (10 mg/kg, starting since the fifth week), respectively. After 4 and 20 weeks of feeding, respectively, serum lipid level and NO concentration were measured in the rats, and the epithelial vascular cell adhesion molecule-1 (VCAM-1) expression and cell adhesiveness to the aortic endothelium were observed.
RESULTSCompared with the control group, the rats with hyperlipidemia induced by long-term high-fat diet feeding showed lower NO concentration and increased leukocyte accumulation on the endothelial surface, exhibiting also stronger and more extensive endothelial expression of VCAM-1. In contrast, the hyperlipidemic rats with fenofibrate treatment shoed significantly decreased VCAM-1 expression and leukocyte adhesion with recovery of the NO level.
CONCLUSIONNO deficiency and activation of inflammation are involved in vascular impairment in rats with high-fat diet-induced hyperlipidemia, and fenofibrate can effectively prevent atherosclerosis by restoring NO concentration and down-regulating VCAM-1 expression in these rats.
Animals ; Atherosclerosis ; prevention & control ; Cell Adhesion ; Endothelium, Vascular ; drug effects ; metabolism ; Fenofibrate ; pharmacology ; Hyperlipidemias ; drug therapy ; metabolism ; Inflammation ; Leukocytes ; cytology ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vascular Cell Adhesion Molecule-1 ; metabolism
4.SNP marker and allele-specific diagnostic PCR for authenticating herbs of Perilla.
Yu-ming LUO ; Wei-ming ZHANG ; Xiao-yu DING ; Jie SHEN ; Shu-lin BAO ; Bi-hai CHU ; Shan-guo MAO
Acta Pharmaceutica Sinica 2006;41(9):840-845
AIMTo authenticate all the varieties of Perilla (single-species genus), to analyze sequences of rDNA ITS regions and single nucleotide polymorphism (SNP) within them and based on these, to design allele-specific diagnostic PCR primers.
METHODSThe rDNA ITS regions of the perilla varieties were sequenced and analyzed by Clustal X 1.8, MEGA 3.0. Allele-specific diagnostic PCR primers that can authenticate all the perilla varieties were designed based on SNPs loci.
RESULTSThe length of rDNA ITS sequences of perilla varieties ranged from 612 to 615 bp in size, including ITS1 (230 -232 bp), 5.8S (179 bp) and ITS2 (203 -204 bp). The GC content is about 61.5% - 61.9%. There is not only SNPs in non-coding region ITS1 and ITS2 (ncSNP), but also three coding SNPs (cSNP) loci in the conservative region of 5.8S. All the SNPs have only two allele loci polymorphism. The cSNP in 5.8S is related to the morphology variation among the varieties. Allele-specific diagnostic PCR primers have been designed according to SNPs loci to authenticate accurately all the seeds and leaves of Perilla varieties.
CONCLUSIONSNPs in rDNA ITS region can be used as an effective molecular markers to authenticate all the varieties of Perilla.
Alleles ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Genetic Markers ; Perilla ; classification ; genetics ; Perilla frutescens ; genetics ; Plant Leaves ; genetics ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; Seeds ; genetics ; Sequence Analysis, DNA ; Species Specificity
5.Surgical therapy for massive deep skin and soft tissue injuries.
Zhao ZHENG ; Da-hai HU ; Ming-da XU ; Xiong-xiang ZHU ; Jun-tao HAN ; Mao-long DONG ; Ke TAO ; Hong-tao WANG ; Song-tao XIE ; Chu-yun JIANG ; Bi CHEN
Chinese Journal of Burns 2009;25(1):11-14
OBJECTIVETo explore the methods of repair of massive deep skin and soft tissue injuries.
METHODSFifty-six patients with deep skin and soft tissue injuries were hospitalized from July 2006 to January 2008. Among them, 23 cases were caused by burn, 17 cases by electric injury, 7 cases by hot crush injury, 6 cases by avulsion injury, and 3 cases due to other reasons (including traffic accident, crush injury, soft tissue infection respectively). Sixty-five skin flaps were raised to repair and reconstruct the injured tissues, including 21 local flaps, 18 distant pedicled skin flaps, and 26 free skin flaps. The area of skin flaps ranged from 1.5 cm x 1.0 cm to 39.0 cm x 23.0 cm.
RESULTSSixty skin flaps survived completely, partial necrosis occurred in 3 flaps, and complete necrosis in 2 flaps. There was no obvious difference in average survival rate among local skin flaps (95.2%), distant pedicled skin flaps (88.8%), and free skin flaps (92.3%, P > 0.05).
CONCLUSIONSSkin flap transposition can be still considered as the major effective method in repair of massive deep skin and soft tissue injury. On the premises of high survival rate, free skin flap transposition can be considered as the first choice.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Burns ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin ; injuries ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; Young Adult