Objective　To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods　The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result　Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion　It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.

Object To provide evidences for the identification of the root of Ampelopsis brevipedunculata (Maxim.) Trautv. var kulingensis Rehd as a basis for the rational exploitation and utilization of this medicinal plant Methods The characteristic features were studied by macroscopic and microscopic observations and its chemical costituents identified qualitatively by TLC Results Macroscopic and microscopic characteristics of this crude drug were described 4 chemical compositions, such as lupeol, were found by TLC Conclusion The distinct characteristics revealed in the studies could provide a basis for the identification of this crude drug

Objective:To explore a new method to identify Sparganium stoloniferum and its adulterants by ITS2 regions. Methods:Eight samples of Sparganium stoloniferum and its adulterants were collected with five species, and 6 species with 23 ITS2 sequence of Sparganium stoloniferum and its adulterants were downloaded from Genbank. The intraspecific and interspecific K2P distances of Spar-ganium stoloniferum and its adulterants were calculated by MEGA5. 0, and the phylogenetic tree was constructed by MEGA 5. 0. Re-sults:The maximum intraspecific K2P distance of Sparganium stoloniferum was 0. 038,while the minimum interspecific K2P distance was 0. 697. The phylogenetic tree showed that Plantago asiatica was different obviously from its adulterants. The different samples of Sparganium stoloniferum were gathered together and could be distinguished from its adulterants by the NJ tree. Conclusion: ITS2 se-quence is able to identify Sparganium stoloniferum and its adulterants correctly, which provides a new method for the identification of Sparganium stoloniferum.

This study was aimed to identify Bletilla Striata (Thunb.) Reichb.f.and Bletilla Formosana (Hayata) Schltr.by ITS2 sequence.The leaves of 38 samples of Bletilla striata and Bletillaformosana from Yunnan,Hubei,Guizhou,Hunan and Sichuan province were used as experiment materials.The total DNA was extracted.Internal transcribed spacer 2 (ITS2) sequences were obtained by PCR.All of the ITS2 sequences were checked.The 8 ITS2 sequences from two species were downloaded from GenBank.The intraspecific and interspecific Kimura-2-parameter (K2P) distances of Bletilla striata and Bletilla formosana were calculated by MEGAS.0.And neighbor-joining (NJ) tree was constructed.The results showed that the full-length sequences of ITS2 from Bletilla striata and Bletillaformosana were 259 bp,with a total of 14 variable sites.The maximum intraspecific K2P distance of Bletilla striata and Bletillaformosana was 0.008,while the minimum interspecific K2P distance was 0.040.The ITS2 secondary structure showed that different origins of Bletilla striata were gathered together and could be distinguished obviously from Bletilla formosana by NJ tree.It was concluded that ITS2 sequence was able to identify Bletilla striata and Bletillaformosana quickly and accurately.

Objective:To observe the effect of Kechuanning on Th cytokine expression in pediatric virus-induced asthma.Methods:120 cases of pediatric virus-induced asthma were randomly divided into Kechuanning group(treatment group,n =60,treated with Kechuanning) and Western medicine group(control group,n=60,treated with virazole).The change of Th1 cytokine IL-12 and Th2 cytokine IL-4 in blood serum were detected and analyzed.Results:Kechuanning could increase IL-12 but decrease IL-4 level in the patient's blood,therefore regulate Th cell subset,and the effect was more obvious than in control group.Conclusion:The mechanism of Kechuanning in curing pediatrics asthma induced by virus was related with increasing IL-12 and decreasing IL-4 level.

OBJECTIVETo construct the two-dimensional electrophoretic (2-DE) protein map of the human sperm head.

METHODSProtein extracts of the normal human sperm and sperm head were loaded on an 18 cm immobilized pH gradients (IPG) strip holder and separated with isoelectric focusing electrophoresis as the first dimension, and then with upright SDS-PAGE as the second. Different protein spots between them were compared with Image Master 3.0, and so were the maps of method I with Urea/Thiourea/Guanidine HCl and method II with Kit/Guanidine HCl.

RESULTSOf the whole-sperm proteome, 802 protein spots were obtained by method I, and 797 by method II. Among them, distribution patterns of 492 spots were the same. A sperm protein map was constructed with 1107 protein spots after the interaction of the spots obtained by both the methods. Of the sperm-head proteome, 428 protein spots were obtained, all found in the whole-sperm protein map after comparison.

CONCLUSIONBoth the methods could be complementarily used to construct a sperm protein map, and the map could serve as a model for the establishment of the sperm protein profile.

Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Hydrogen-Ion Concentration ; Male ; Proteins ; analysis ; Proteomics ; methods ; Sperm Head ; chemistry

OBJECTIVETo determine the candidate sequences which can be used as DNA barcode to identify species in Caprifoliaceae family by screening out from four different DNA fragments sequences.

METHODPCR amplification, sequencing efficiency, differential intra- and interspecific divergences, the DNA barcoding gap and identification efficiency were used to evaluate these loci.

RESULTThe ITS2 was used as a candidate sequence of DNA barcode to identify the species in Caprifoliaceae family, whose rate of success in identification in genera level was 100% and in species 96.6%, and psbA-trnH as a complementary barcode to ITS2 for Caprifoliaceae.

Automatic Data Processing ; methods ; Base Sequence ; Caprifoliaceae ; genetics ; DNA ; analysis ; DNA, Plant ; analysis ; Identification (Psychology) ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; Species Specificity