Objective To explore the application probability of AFLP method (Amplified Fragment Length Polymorphism) for the genetic study of complicated samples and to make clear the analytical approach on polymorphic fragments.Methods An AFLP fragement sequence,resolved between murines with high and low metastatic hepatocarcinoma,was retrieved from a reference paper and was annotated on NCBI database by different BLAST programs. Results The retrieved information varied along with the different BLAST programs, nevertheless BLASTX program provided more comprehensive information and showed that the differentiated fragment exhibited high homology with terminase large subunits from Burkholderia phage Bups phil and Mycobacterium tuberculosis H37Ra. However, there is no detailed information for the latter,key words search returned the relationship between terminase and the formation and metastasis of tumors and provided the further research clues. Conclusions Whole genome screening in parallel of AFLP markers between genetically differentiated paired materials would readily produce informatic fragments. Further analysis of the sequenced fragments by informatic methods would facilitate the subsequent research.

Aim To study the application status of odds ratio for medical animal experiments.Methods Odds ratio and seven kinds of animal models were used as retrieval strategy to search medical animal experiment related papers in several Chinese and English databases.Papers relating to each kind of animal model and using odds ratio in abstract and text were counted. Data from different databases were compared. Calculation of odds ratio was exemplified and the significance of different odds ratio values was illustrated in this paper.Results Few medical animal experiments cited odds ratio as statistics.Conclusions The importance of odds ratio has not been fully recognized in Chinese references.

Objective:To study the cellular pathology and molecular mechanisms of intestinal mucosal damage induced by IgG immune complex in mice.And to explore the pathogenic mechanism and molecular diagnosis evidence of ulcerative colitis induced by food intolerance in clinical practice.Methods: Six weeks old BALB/c female mice were used to build animal model.All the mice were divided into four groups:the control group(group A),the rabbit intestinal mucosal protein immunized group(group B),the DSS induced group(group C),the rabbit intestinal mucosal protein immunized combined with DSS induced group(group D).After successful establishment of animal model,serum and colon tissues were collected to be performed relevant tests.Results: IgG level in serum and colonic mucosa of group B mice increased.And inflammatory cell infiltration and a small amount of mast cell activation were observed in intestinal mucosa;group C mice showed the typical acute ulcerative colitis:a large number of inflammatory cells infiltration,inflammatory factor levels increased in mucosa and mucosa lamina propria,and mucosal epithelial cells′ tight junction weakened;group D mice manifested both high level of IgG in serum and colonic mucosa and also typical acute ulcerative colitis.Besides,significant mast cell activation was observed in the intestinal mucosa.Conclusion: We infer from the experimental results that IgG immune complexes can induce the damage of intestinal epithelium by mediating activation of mast cells.And during the process,the level of inflammatory cytokines increased in intestinal mucosa and the expression of tight junction protein in epithelial cell decreased.These factors contribute to the promotion of intestinal mucosa damage induced by immune complexes.

Objective To study the localization of specific allergen of Dermatophagoides pteronyssinus. \ Methods\ Through optical microscope,the specific allergens of D.pteronyssinus were observed in paraffin sections using D.pteronyssinus\|specific IgE antibodies from the patient sera. \ Results and Conclusion \ The digestive system was found occupying large parts of body cavity of D.pteronyssinus by HE staining, while the specific allergens of D.pteronyssinus were mostly occurred in the midgut tissue, gut contents, cuticle and reproductive system in the immunostained sections. The results also showed that many parts of D. pteronyssinus were recognized by the specific IgE antibodies obtained from allergic individuals to D.pteronyssinus, which provided a theoretic base for further study of isolation and purification of the specific allergen.

Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.

Objective To compare the efficacy of thrombus aspiration catheter combined with intracoronary tirofiban and nitroglycerol injection through the aspiration catheter versus the guiding catheter during primary percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI).Methods Thirty-four patients with STEMI undergoing primary PCI and receiving thrombus aspiration catheter combined with intracoronary tirofiban and nitroglyeerol injection through the aspiration catheter were enrolled as the aspiration group (n =34),and those who had similar coronary angiography results and basic characteristics but receiving thrombus aspiration catheter combined with intracoronary tirofiban and nitroglycerol injection through the guiding catheter were served as the guiding group ( n =33 ).The outcomes of the two groups were observed and compared.Results There was no significant change of blood pressure between before and after injection in the aspiration group ( P ＞ 0.05 ),but the change of blood pressure was significant after injection compared with before injection in the guiding group ( P ＜ 0.01 ).The cTn-I,BNP,peak-value of CK-MB,peak-time of CK-MB,TIMI grade 3 flow,slow-reflow in IRA after PCI in the aspiration group were superior to those in the guiding group ( t =3.92,P ＜ 0.01 ;t =4.70,P ＜ 0.01 ; t =3.39,P ＜ 0.01 ; t =7.17,P ＜0.01 ; x2 =3.877,P ＜ 0.05 ; x2 =3.876,P ＜ 0.05 ).LVEF,LVEDd and LVESd after 1 month in the aspiration group were superior to those in the guiding group (t =5.99,P ＜ 0.01 ;t =4.53,P ＜ 0.01 ;t =8.12,P ＜ 0.01 ),but no significant differences of LVEF,LVEDd,LVESd were found after 1 week resolution of sum of ST-segment elevation and the MACE rates after PCI were found between the two group ( P ＞ 0.05 ).Conclusion Application of thrombus aspiration catheter combined with intracoronary tirofiban injection through the aspiration catheter is more effective than through the guiding catheter in patients with Acute ST-segment elevation myocardial infarction,which could decrease slow-reflow phenomenon and improve re-perfusion and left ventricular function with better clinical outcomes.

Objective:To obtain purified recombinant Litopenaeus vannamei allergen protein Lit v 1.2.Methods: The target gene of Lit v 1.2 was inserted into clone vector pGEM-T and then ligated to the expression vector pET 44a.The pET44a-Liv 1.2 was transformed into Rosetta and screened by ampicillin resistance .The recombinant protein was expressed by IPTG induction .The protein was purified by 6-His tag affinity chromatography and the purification was analyzed by SDS-PAGE gel electrophoresis .Results:The ex-pression plasmid pET44a-Lit v 1.2 was constructed.SDS-PAGE showed that expressed Lit v 1.2 was efficient and soluble in E.coli Rosetta.The protein molecular weight was consistent with the theoretical value .The highly purified target protein was obtained.Conclusion:In this study ,we successfully gained highly purified recombinant allergen protein Lit v 1.2 which was expressed in prokaryotic system and purified by affinity chromatography column .The purified Lit v 1.2 protein will facilitate us to further study its role in immunological responses .

Objective:To analyse the physicochemical properties and structure of major privet pollen allergen Lig v 1 using bioinformatics software and provide a reference for choosing suitable recombinant expression system for Lig v 1 and modifying the allergen Lig v 1 experimentally.Methods:The physicochemical properties were analysed by ProtParam,the signal peptide by SignalP 4.1 Server,the transmembrane helix by TMHMM Server v.2.0,the secondary structure by GOR4,MHCⅡepitopes by NetMHCⅡ2.2 Server,B-cell epitopes by ProteanTM 5.01 and the phylogenetic tree by MEGA 6.Results: Privet major pollen allergen Lig v 1 was stable in Escherichia coli and it doesn′t possess any signal peptide and transmembrane helix.Most secondary structures of Lig v 1 were random coils.Potential region of MHCⅡepitope of Lig v 1 was 30-44.Potential B-cell epitopes possess discontinuous and continuous a mino acid sequences.Lig v 1 and its counterparts from Fraxinus excelsior and Olea europaea were clustered into one group.Conclusion:Escherichia coli is the suitable expression system for recombinant Lig v 1.In silico prediction of the epitopes of Lig v 1 provides a reference for modifying the allergen Lig v 1 experimentally.

Objective To evaluate the safety and clinical efficacy of stent implantation using a rapid artificial cardiac pacing technique on ostial lesions of left anterior descending artery (LAD).Methods From Jun 2008 to Nov 2010,38 patients with ostial lesions of LAD were recruited and randomly divided into two groups:patients with stent implantation using a rapid artificial cardiac pacing technique (pacing group,n =19 ) and patients with direct stent implantation (no-pacing group,n =19 ).Post-stenting examination was performed.Patients were followed-up for 9 months and coronary angiography was reviewed.The immediate success rate,major adverse cardiac events including death,reinfarction and target vessel revascularization,late lumen loss,sent thmmbosisin,rent-restenosis were compared between these two groups.Results There were no significant differences in the baseline values,disease characteristics and instant response to surgery between pacing and no-pacing groups(P ＞ 0.05 ).The time cost for stent placement was significantly shorter in the pacing group than that in the no-pacing group ( [ 16.5 ± 0.5 ] s vs.[46.6 ± 1.4 ] s,t =88.256,P =0.004 ).After surgery,there was no acute or subacute thrombosis,in-stent restenosis or occlusion for patients in the pacing group.In the no-pacing group,one patient developed acute thrombosis.The symptoms disappeared after thrombus aspiration and balloon dilatation by emergency percutaneous coronary intervention.Patients were followed up for 270 -275 days,and patients in the pacing group received post-stenting coronary angiography 9 months after stent implantation using a rapid artificial cardiac pacing technique,and no in-stent re-stenosis was found.During this period,nobody had adverse events such as death,myocardial infarction or target lesion revascularization,while five cases with in-stent restenosis (50％ -60％ )were found in the no-pacing group,without further target lesion reconstruction due to symptomless.Conclusion Compared with previous positioning technique,stent implantation using a rapid artificial cardiac pacing technique on treatment of ostial lesions of LAD is safer,and more effective.It is a favorable method for accurate positioning of bracket and can improve the prognosis,reduce the occurrence of acute thrombosis and in-stent restenosis.

Objective To observe the safety and efficiency of DIVER thrombus aspiration catheter application during percutaneous coronary intervention(PCI) in patients with acute myocardial infarction(AMI)and to evaluate its impacts on the myocardial reperfusion,cardiac function and in-stent restenosis after 9months.Methods A total of 86 cases of ST-segment elevation patients of AMI treated with PCI and confirmed complete occlusion lesion by angiography from November 2008 to December 2010 were randomly divided into two groups:aspiration group(n =43)with DIVER thrombus aspiration cathetcrs were used,and non-aspiration group (n =43).ST-segment recovery within two hours,TIMI grade,the in-hospital adverse major cardiac events,the levels of Pro-BNP,coronary function determined by ultrasound heartbeat graph and the results of coronary angiography after PCI for 9 months were compared between two groups.Results There was significant difference on ST-segment recovery within two hours between these two groups[95.35％(41/43)vs 79.02％(34/43),x2 =11.862,P =0.0006].The incidence of TIMI 3 grade flow was significantly higher,the incidence of TIMI 2(slow flow)and TIMI 0-1(no-reflow)grade flow were much lower in aspiration group than those in non-aspiration group immediately after PCI[TIMI 3:93.02％(40/43) vs 81.40％(35/43),x2 =6.06,P =0.0335 ; TIMI 2:6.98％(3/43) vs 13.95 ％(6/43),x2 =3.12,P =0.0495 ; TIMI 0-1:0 vs 6.98 ％(3/43),x2 =5.29,P =0.0352].There was no difference on LVEF[(0.420±0.054) ％ vs(0.408±0.052)％,t =1.0496,P =0.766and LVEDD(56.5±4.5)mm vs(57.6±4.4)mm,t =1.0419,P =0.7832]between these two groups one day after PCL LVEF was significandy higher in aspiration group than that in non-aspiration group one and nine months after PCI[(0.452±0.050) ％ vs(0.432±0.049) ％,t =3.3957,P =0.0482 ;(0.469±0.053) ％ vs (0.413±0.052)％,t =4.9457,P =0.0336].LVEDD was significantly smaller in aspiration group than that in non-aspiration group one and nine months after PCI[(49.6±5.1) mm vs(53.4±4.6) mm,t =3.4548,P =0.0473 ;(46.5±4.4) mm vs(50.2±4.8) mm,t =3.7260,P =0.0421].There were no cardiovascular events (angina,myocardial infarction,heart failure,cardiac death)in both groups during 9 months follow-up.The coronary angiography results showed that the occurrence rate of in-stent restenosis in aspiration group was significantly lower than that in non-aspiration group(2.33％ vs 6.98％,x2 =4.4351,P =0.0463).Conclusion The application of DIVER thrombus aspiration catheters during PCI in all patients with AMI was safe and effective,it can improve the myocardial tissue perfusion and the post-operative cardiac function and can lower the occurrence rate of in-stent restenosis.