Objective To construct the multi-epitope antigen(mea) gene of G2 glycoprotein of Hantavirus SEO type L99 strain.Methods The B cell epitopes was chosen and connected by three-peptide GPG after the amino acid sequence of G2 protein was analyzed and predicted by bioinformatical soft wares.The corresponding gene mea was constructed by overlap PCR,and cloned into the prokaryotic expression plasmid pET32a(+).Results The mea gene was successfully constructed by the five epitopes being chosen.The recombinant plasmid pET32a-mea was acquired by directional cloning.Conclusion The mea and its expression system E.coli BL21/pET32a-mea were constructed for the first time.The foundation was laid for the expression of the mea and its immunological applications.

Objective To evaluate the effectiviness and safety of rituximab in the treatment of immune thrombocytopenia .Meth-ods Standard doses and small doses of rituximab alone or combined therapy with other methods were used in 2 cases with immune thrombocytopenia who diagnosed and treated for the first time ,and 6 cases who diagnosed with refractory recurrent immune throm-bocytopenia .The recent curative effect ,adverse reactions were analyzed .Results The recent curative effect was well in the 2 cases who diagnosed and treated for the first time ,but 1 case recurrenced after 6 months .4 cases of effective in the 6 cases who diagnosed with refractory recurrent ,in which 1 case recurrenced after 2 years ,2 cases had no effect .All of the patients have no obvious adverse reaction in the near future .Conclusion Rituximab treatment of immune thrombocytopenic ,it is of good curative effect ,good safety and tolerability .

Objective To study the role of adiponectin (ADP)post-conditioning in alleviating myocardial ischemia reperfusion injury (MIRI) in 3-week old rats.Methods SD rats (3-week old) were randomly divided into 4 groups by random digits table (n =6):sham group was processed identically except that the left anterior descending coronary artery(LAD) was not ligated;MIRI group was subjected to occlusion of the LAD followed by reperfusion,occlusion for 30 min and reperfusion for 120 min;ADP group was subjected to injection of ADP into femoral vein before reperfusion;LY294002 group (ADP post-conditioning and LY294002) was subjected to injection of ADP and LY294002 into femoral vein before reperfusion.The titer of lactate dehydrogenase(LDH),cardiac troponin I(cTnl) and malondialdehyde(MDA) in plasma were observed.The levels of superoxide dismutase (SOD)and MDA in myocardial tissue were observed.Results The levels of LDH,cTnI and MDA in plasma of MIRI group was (732.11 ± 111.29) IU/L,(38.05 ± 6.17) g/L and (4.49 ± 0.17) nmol/L,respectively,compared with sham group,the levels of LDH,cTnI and MDA in plasma of MIRI group increased obviously(all P ＜ 0.05);the levels of LDH,cTnI and MDA in plasma of ADP group were (581.72 ± 68.41) IU/L,(20.70 ± 5.43) g/L and (3.10 ± 0.20) nmol/L,respectively;compared with MIRI group,the levels of LDH,cTnⅠ and MDA in plasma of ADP group declined significantly (all P ＜0.05);the levels of LDH,cTnI and MDA in plasma of LY294002 group were (701.10 ±99.59) IU/L,(33.13 ± 4.32) g/L and (4.19 ± 0.46) nmol/L,compared with ADP group,the levels of LDH,cTnI and MDA in plasma of group LY294002 increased remarkably (t =2.477,1.804,2.961,all P ＜ 0.05).Compared with sham group,the SOD level decreased and the MDA level increased in myocardial tissue of MIRI group(all P ＜ 0.05);compared with MIRI group,the SOD level increased and the MDA level decreased in myocardial tissue of ADP group (all P ＜ 0.05);compared with ADP group,the SOD level decreased and the MDA level increased in myocardial tissue of LY294002 group (all P ＜ 0.05).Conclusions ADP postconditioning is perhaps a protective factor for alleviating myocardial ischemia reperfusion injury in 3-week-old rats,the protective effect is perherps related to alleviating the damage of the lipid peroxidation by signaling pathway of adiponectin/phosphoinositide-3 kinase/protein kinase B.

Objective To investigate the genetic diversity of five populations of cultivated Morinda officinalis in Guangdong Province using random amplified polymorphic DNA(RAPD).Methods Sixty-four individuals in five populations of cultivated M.officinalis were analyzed by RAPD markers to determine the genetic variations among the populations.The data of genetic diversity were analyzed with Popgen32 software.Results The levels of genetic variation and patterns of population structure in M.officinalis were investigated using RAPD markers.Of the 100 primers screened,15 primers produced highly reproducible RAPD bands,using these primers,224 discernible DNA fragments were generated with 112(50.00%) polymorphic fragments,indicating considerable genetic variation at the species level.In contrast,there were relatively high levels of polymorphism at the population level with the percentage of polymorphic bands ranging from 37.05% to 53.13%,and the mean percentage of polymorphic loic(P=45.86%).Genetic variation among populations was detected based on Nei's genetic diversity analysis(0.175 6),Shannon's diversity index(0.287 6) for every population with Shannon's index 0.103 3—0.236 2 and Nei's indexes 0.074 5—0.154 0.Conclusion There is a little genetic differentiation among populations of cultivated M.officinalis and result in different cultivated types,the genetic diversity with M.officinalis(POP2) is higher than that of other populations.It may be the main reason of the difference of cultivated M.officinalis quality.

Objective To summarize radiofrequency cather ablation (RFCA)of tachycardias with right sid-ed accessory atrioventricular pathway in 7 patients with persistent left superior vena cava (PLSVC). Methods RF-CA for right sided accessory atrioventricular pathway in 7 patients with PLSVC was carried out . Coronary sinus ac-cess was performed through the left subclavian and the left femoral vein respectively. Swartz sheath was used during ablation if neccesary. Results There was no siginificant differences in fluoroscopy time between the left subclavian approach and the left femoral vein approach[(2.8 ± 0.6)min vs (3.2 ± 0.3)min, P>0.05)]. No recurrence of ac-cessory pathway conduction was found during a mean follow-up of (23 ± 11) months. The mean discharging time and fluoroscopy time were (312 ± 67)s and (17 ± 5)min respectively. Conclusions Although RFCA of right sided accessory atrioventricular pathway in patients with PLSVC shows specialities in cather manipulation , mapping and ablation, high successful results can be achieved by experienced physician.

Objective To investigate the diagnostic value of T-SPOT.TB in patients with fever.Methods T-SPOT.TB was carried out in 456 febrile patients who had no determined reason for fever or who could not be excluded from infection with Mycobacterium tuberculosis.These patients were followed up for their final clinical diagnosis and the diagnostic value of T-SPOT.TB was evaluated using the receiver operating characteristic (ROC) curve.Results In the present study,85 patients were clinically diagnosed with tuberculosis (TB) (including 34 cases of pulmonary TB,24 cases of TB infection defined as T-SPOT.TB positive without TB lesion,but effective with anti-TB treatment,10 cases of tuberculous pleurisy and 17 cases of other extrapulmonary TB) and 371 patients had no definite evidence of Mycobacterium tuberculosis infection (including 42 cases of suspected TB and 329 cases of other diseases).T-SPOT.TB had a high sensitivity of 84.3％ in TB patients and a high negative predictive value of 94.2 ％,but the specificity was 68.1％ and the positive predictive value was 41.4 ％.The Kappa value was 0.380 and the area under ROC curve was 0.762 (95％CI:0.707-0.817).T-SPOT.TB had a higher sensitivity in extrapulmonary TB than in pulmonary TB (88.2 ％ vs 78.1 ％),with the negative predictive values of 95.3 ％ and 92.6 ％,Kappa values of 0.438 and 0.299,and the areas under ROC of 0.791 (95 ％ CI:0.724-0.857) and 0.719 (95％CI:0.623-0.815),respectively.Ninety-nine cases were T-SPOT.TB positive out of 329 patients diagnosed with other diseases.Conclusions T-SPOT.TB test can be used as a method for TB diagnosis in patients without obvious TB lesions.Due to its relatively high negative predictive value,T-SPOT.TB can be used to rule out infection with Mycobacterium tuberculosis.

Objective To study the role of adiponectin (ADP) postconditioning on myocardial ischemia reperfusion injury (MIRI) in rats .Methods SD rats were divided randomly into 3 groups :⑴Group Sham were processed identically except the left anterior descending coronary artery(LAD) and was not ligated;⑵ Group MIRI were subjected to LAD occlusion for 30 min and reperfusion for 120 min;⑶ Group ADP were subjected to ADP injection before reperfusion .The titer of lactate dehydrogenase (LDH) ,cardiac troponin I(cTnI) and malondialdehyde(MDA)in plasma were observed .The levels of superoxide dismutase(SOD) and malondialdehyde(MDA)in myocardial tissue were observed .The results of HE staining and electrocardiograms(ECG) was ob‐served .Results Compared with Group Sham ,LDH ,cTnI and MDA in plasma in Group MIRI increased (P<0 .05);Compared with Group MIRI ,LDH ,cTnI and MDA in plasma in Group ADP declined (P<0 .05);Compared with Group Sham ,SOD decreased and MDA increased in myocardial tissue in Group MIRI (P<0 .05);Compared with Group MIRI ,SOD increased and MDA decreased in myocardial tissue in Group ADP (P<0 .05) .The myocardial injury was serious in Group MIRI while it was lightened in Group ADP .There were acute ischemia ECG change in Group MIRI while it was slight in Group ADP .Conclusion ADP postconditioning is a protective factor against MIRI in rats ,it is related to eliminating free radicals and alleviating the damage of lipid peroxidation .

Objective To compare the treatment outcome of underwent surgery plus radiotherapy and radiotherapy/chemoradiotherapy plus salvage surgery for stage Ⅲ/Ⅳ laryngeal carcinoma,to investigate an optimized treatment for the patients of stage Ⅲ/Ⅳ laryngeal carcinoma.Methods Clinical data from 103 patients with stage Ⅲ (39 cases) or stage Ⅳ (64 cases) laryngeal carcinoma were retrospectively analyzed.The patients were divided into surgery plus radiotherapy group (S±R,46 cases) and radiotherapy/chemoradiotherapy plus salvage surgery group (R±S,57 cases).Overall survival,relapse free survival,and laryngeal preservation rate were used to compare the treatment outcome between two groups.Multivariate regression models were used to analyze the independent factors for survival and laryngeal preservation rate.Results Survival rate was higher in S±R group than in R±S group [2 year overall survival/relapse free survival 74.7 ％ (34/46) / 72.4 ％ (33/46) vs 46.4 ％ (26/57) / 40.9 ％ (23/57),P ＜ 0.05].Laryngeal preservation rate was higher in R±S group than in S±R group [93.0 ％ (15/46) vs 32.6 ％ (53/57),P ＜ 0.05].Multivariate analysis demonstrated that treatment modality and T stage were independent factors for long-term survival,while treatment modality was the only an independent factor for laryngeal preservation rate.Conclusions Surgery plus radiotherapy result in better survival and lower laryngeal preservation rate than radiotherapy/chemoradiotherapy plus salvage surgery in treatment of stage Ⅲ/Ⅳ laryngeal carcinoma.Surgery plus radiotherapy should be the first choice for treatment of locally advanced laryngeal carcinoma.Improvement of the quality of life could be achieved by laryngeal preservation surgery and phonation reconstruction procedures.

Objective To study proteome variation between uropathogenic E. coil (UPEC)132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. Methods Two-dimensional gel electrophoresis(2-DE) was applied to compare the differential expression proteins between UPEC 132, UPEC J96 and non-uro-pathogenic E.coli K-12 MG1655. The differential expression proteins were digested in gel by enzyme. The mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were re-searched using the internet available database. Results The number of protein spots recognized from UPEC 132 was 466±11, significantly more than that of E. coli K-12 MG1655 (338±15) and UPEC J96 (382±12); there were 298 protein spots shared by the three E.coli strains, 56 protein spots shared by two UPEC strains, and 89 protein spots characterized by UPEC 132. Twenty-two differential expression or significantly increased expression protein spots, involved in virulence factors, metabolism and transportation, regulation of protein synthesis, biological oxidation and unknown functions, were successfully identified by MALDI-TOF-MS. Condusion The proteome from UPEC 132 and non-uropathogenic E. coli K-12 MG1655, or UPEC 132 and UPEC J96 was differentially expressed. It will provide important information on the pathogen-esis of UPEC 132.

Objective To study the effect of low-dose dexamethasone after acute and subacute cerebral infarct.Methods One hundred and forty patients with acute ischemic stroke were randomly divided into acute ischemic stroke group (acute stage group),subacute ischemic stroke group (subacute stage group),placebo and control groups.Subjects in acute stage groups received conventional therapy and 1 mL (5 mg) dexamethasone injection from 1 day to 3 day after admission and in subacute stage group,received the same treatment as acute stage group from 4 day to 6 day after admission.In control and placebo groups,subjects received conventional therapy and conventional therapy + 1 mL normal saline for injection respectively.One week after treatments,complete blood count and erythrocyte sedimentation rate were tested.One month and three month after treatments,neurological function were evaluated by Barthel Index and modified Rankin Scale (mRS).Results The valuate of erythrocyte sedimentation rate in acute stage group were significantly different from it of placebo and control groups (P ＜ 0.05).Moreover,neurological function of subjects in acute stage group was significantly improved than that in placebo and control groups in by Barthel Index and mRS (P ＜ 0.05).However that in subacute stage group was not different from that placebo and control groups (P ＞ 0.05).Conclusion Low-dose dexamethasone plays a neuroprotection role after acutc cerebral ischemia.