Objective To develop a DNA microarray for HLA-DR1,DR51 group genotyping. Methods According to the specific allelic sequences coding HLA-DR1,DR51 loci,HLA- DR1,DR51 group typing probes which were immobilized on a glass support were synthesized.A pair of group-specific primers labeled by the Cy5-dCTP were designed,then the primers were used in the PCR,thus the PCR products were labeled with Cy5.The labeled PCR products were hybridized with array.The signals were scanned by scanner and analyzed by image software.The typing results were confirmed by standard DNA and PCR-SSO. Results A total of 130 samples were typed by this DNA array.There were 34 HLA-DR1,DR51 group loci typed by DNA array.Among them,18 loci were DR15,8 were DR16,6 were DR10 and 2 were DR1.No false positive or false negative typing results occurred.The accuracy and reproducibility were 100% and the overall time of genotyping was about 3.5 hours. Conclusions DNA array technique is a precise,rapid molecular method of high resolution power and high specificity for HLA-DR1,DR51 genotyping,which is applicable to clinical transplant practice.

Objective To develop an oligoneucleotide array for HLA-DR52 group rapid genotyping.Methods According to the special allele sequences of HLA-DRB loci in Chinese Han's population, HLA-DR52 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed and were used in the PCR. The labeled PCR products with Cy5 were hybridized with array. The signals were scanned by a scanner and analyzed by Image software. 83 samples were typed by this array and the results were compared with PCR-SSP typing.Results Among 57 HLA-DR52 group loci typed by PCR-SSP,2 samples had no HLA-DR52 loci typed by array,3 DR52 group homozygotes typed by PCR-SSP were actually heterozygotes by array. The other 1 non-DR52 group homozygote identified by PCR-SSP was a heterozygote with one DR52 group locus. Conclusion The oligoneucleotide array technique is a precise, rapid molecular method for HLA-DR52 genotyping. Compared with PCR-SSP method, the genotyping chip is more sensitive and intuitionistic and suitable for clinic practice.

Objective To study the possible mechanism of Bushenqiangdu recipe in treatment of ankylosing spondylitis(AS). Methods 30 out-patients were treated by Bushenqiangdu recipe for 6 months. The expression level of IL-18,IFN-?,IL-4 mRNA in outer-circumference blood of as patients and healthy volunteers were studied,as well as the change expression level of IL-18,IFN-?,IL-4 mRNA of as patients outer-circumference blood before and after treatment. Result The expression level of IL-18,IFN-?mRNA in AS patients PBMC group is higher prominently than control PBMC group(P

Objective To investigate the effect of resveratrol on high glucose-induced oxidative stress and cardiomyo-cyte hypertrophy and possible mechanism .Methods H9C2 cardiomyocytes were divided into normal glucose group , high glucose group , resveratrol group and resveratrol +DAPT group respectively .The cell surface was measured by rhodamine-labeled phalloidin .Reactive oxygen species generation was measured using DCFH-DA.The contents of SOD and MDA were evaluated by colorimetry .The mRNA and protein levels of Notch 1, Hes1, ANP and BNP were evaluated by RT-qPCR and western blot .Results High glucose exposure significantly increased cell surface area ( P<0.01) and enhanced ROS and MDA generation ( P <0.01 ) with concomitant decrease of SOD activity ( P <0.01).It also decreased the mRNA and protein expressions of Notch 1 and Hes1(P<0.05), and increased the ex-pression levels of ANP and BNP ( P<0.05 ) .Resveratrol treatment inhibited cell surface expanding ( P<0.01 ) and decreased ROS and MDA generation (P<0.01) with concomitant enhance SOD activity (P<0.01), as well as in-creased the expression level of Notch 1 and Hes1 ( P<0.05 ) , and decreased the expression of ANP and BNP ( P<0.05).Conclusions Resveratrol may inhibit high glucose induced H9C2 cardiomyocytes hypertrophy by improving oxidative stress and activating Notch 1/Hes1 signaling pathway .

Objective To explore the regulating mechanism of inducible nitric oxide synthase(iNOS) in hepatic injury of obstructive jaundice (OJ) in vivo and in vitro experiments. Methods (1) Rat hepatocytes were isolated by in situ collagenase perfusion and primary culture. Hepatocytes were pretreated with various concentrations of iNOS inhibitor SMT for 20 min. After pretreatment, 50?M GCDC was added for an additional 24hr. Cells were next detected by FCM and TUNEL.(2) Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct in rats. After BDL for 3d、7d、14d、and 21d, the apoptotic status in liver of all rats were determined with TUNEL, and iNOS protein in liver of OJ was ditermined with immunohistochemistry method. Results (1) SMT decreased GCDC-induced apoptosis in a concentration-dependent manner. (2) The apoptotic rate of liver was related to length of time of OJ. Apoptosis index (AI) was highest from rats with 14d bile duct ligation. The stronger the iNOS expression, the higher was the number of apoptotic cells that was found in OJ. Conclusions iNOS is involved in the regulation and the occurrence and progression of hepatic injury of obstructive jaundice.

Objective To investigate the distribution and changes of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in normal heart or heart after myocardial infarction (MI),and to explore beneficial effects of bFGF on heart after MI. Methods Eleven adult minipigs were randomly divided into 2 groups,control (n=5) and MI group (n=6). MI was induced by ligating the left anterior descending coronary artery. Sham-operation was carried out on the animals of control. bFGF-like-immunoreactivity (bFGF-ir) and FGFR-1-like-immunoreactivity (FGFR-1-ir) in heart tissues were detected by immuno-histochemistry and image analysts in 4 weeks after surgery. Results In controls,bFGF-ir and FGFR-1-ir in the atrium showed a considerable high level compared with 2 ventricles (P

Horizontal gene transfer (HGT), also Lateral gene transfer (LGT), is any process in which an organism transfers genetic material to another species that is not its offspring. With the increase of available genomic data, it has become more convenient to study the way to detect the genes, which are products of horizontal transfers among a given genome. There are few data about known horizontal gene transfers in three bacterium genomes under consideration, so the experiments, which simulated gene transfer by artificially inserting phage genes, were carried out. Combining the feature analysis methods of gene sequences with support vector machine (SVM), a novel method was developed for identifying horizontal gene transfers (HGT) in 3 fully sequenced bacterium genomes (Escherichia coli K12, Borrelia burgdorferi, Bacillus cereus ZK). According to our previous work, codon use frequency (FCU) was selected as the sequence feature, in respect that it is inherently the fusion of both codon usage bias and amino acid composition signals. In addition, another computational method was proposed considering strand asymmetry and predicting horizontal gene transfers of leading strand and lagging strand of genomes under consideration, respectively. To avoid the occasionality of simulating gene transfer through artificially inserting phage genes, 100 times of the transfer-and-recover experiment were repeated and arithmetic average of measurement for each genome being considered were reported to evaluate algorithm's performance. Ten-fold cross-validation was used for both parameter and accuracy estimation. The best results were obtained for C-Support Vector Classification (C-SVC) type by using the radial basis function kernel with ?=100, while for one-class SVM type the best performance was obtained using the polynomial kernel of three degree. The performance of the approach was compared with that of Tsirigos' method ,which is one of the best predictive approachs to date in detecting of horizontal transfer genes. Firstly, for the original method that did not consider the strand asymmetry, the C-SVC type has a high relative improvement(RI) of 31.47% on hit ratio for Escherichia coli K12, while the one-class SVM type has RI of 11.61% for Borrelia burgdorferi. Moreover, as theoretically expected, the method considering the strand asymmetry resulted in higher RI than the original method. In order to examine the approach's performance in detecting factual gene transfer events, the approach was applied in genome of Enterococcus faecalis V583. It is not only succeed in recovering all the seven factual horizontally transferred genes, also found that the whole segment from 7 kb upstream of gene EF2293 to 38 kb downstream of gene EF2299 was probably transferred into E. faecalis V583 genome simultaneously with the above seven genes.

AIM: To observe the curative effect of titanium wiring in treatment of the fractures of Neer Ⅱ type distal clavicle and dislocation of Tossy Ⅲ type acromioclavicular joint. METHODS: From March 2004 to December 2006, 37 cases with Neer Ⅱ type distal clavicle fractures or Tossy Ⅲ type acromioclavicular joint dislocation were treated with titanium wiring in Changzheng Hospital, Second Military Medical University of Chinese PLA. Titanium wiring (produced by Sofamor-Danek Company in USA) had good histocompatibility, intensity of tension and flexibility. The broken coracoclavicular ligament was repaired directly by titanium wiring in all cases. The outcomes were evaluated according to Herscovici's criteria. RESULTS: All the cases were followed up for 6 to 24 months. All the dislocated acromioclavicular joints returned to normal 2 weeks after operation. Excellent and good rate of shoulder function was 100% according to Herscovici standard. X-ray imaging showed all clavicle fractures united, acromioclavicular joint had no redislocation or titanium wiring breaking. The incisions of all cases had no obvious swelling and exudation. CONCLUSION: When Neer Ⅱ type distal clavicular fracture and Tossy Ⅲ type acromioclavicular joint dislocation are treated with titanium wiring, the acromioclavicular joint can exercise earlier and recover quickly.

Objective To screen a kind of microorganism which is low poisonous for living creature but has high molluscicidal effect. Methods The Xanthobacter autotrophicus from the soil where Oncomelania hupensis lived were detached, then its molluscicidal effect against the snails were studied by the immersing and contacting methods, respectively. Results The death rates were 1.3%-95.0% immersed for 24 to 72 hours in the different density liquids (1?106,2?106,3?106,4?106 cfu/ml). The LD_~50 immersed for 48 hours was 2.6?106 cfu/ml. The death rates had obviously positive relationship with the immersing time and the density of the liquid. Conclusion The liquid which contains more than 106 cfu/ml Xanthobacter autotrophicus has a stable molluscicidal capacity, and the immersing method is better than the contacting method.[

Objective To analyze molecular evolution of carbapenemase KPC-12 and its binding free energies with β-lactams.Methods Class A beta-lactamases were divided into 2 clusters:those with carbapenemase activities and those without.Minimum Evolution method in MEGA4.1 software was used to analyze molecular evolution of class A beta-lactamases with carbapenemase activity,including KPC-2 to KPC-13,SFC-1,SME-1,IMI-1,NMC-A,and class A beta-lactamases without carbapenemase activity,including TEM-1,SHV-1.Then,tertiary structure of KPC-12 was predicted by homology modeling as reported in SWISS-MODEL database depending on tertiary structure of KPC-2.Moreover,DOCK module in ArgusLab 4.1 software was used to perform molecular docking of KPC-12 to 10 kinds of beta-lactams substrates,and the binding free energies (△ G) were calculated.Results Molecular evolution between KPC-12 and KPC-2 was the closest.The top three decline in binding free energies of β-lactams were penicillin G sodium salt (△G =-8.45149 kcal/mol),ertapenem (△G =-8.36383 kcal/mol) and ampicillin (△G =-8.19326 kcal/mol),while the last two decline in binding free energies of β-lactams were aztreonam (△G =-6.50614 kca]/mol) and clavulanic acid (△G =-6.88533 kcal/mol).Conclusion Carbapenemase KPC-12 has high catalytic activities to penicillin G sodium salt,ertapenem and ampicillin,while has low catalytic activities to aztreonam and clavulanic acid.