According to the results of quantum chemistry calculation and the present research status in the relationship between the structures and the functions of DT, the E154 in DT catalyzing domain was mutated to aspartic acid and arginine in order to study the effects of the alteration on the biological activities. By means of gene site-direct mutation, two mutated genes were prepared and the high performance expression was obtained in E.coli system. The results of toxcity studies indicated that the acute toxicity in guinea pig and cytotoxicities of mutant E154D increased slightly in compared with those of recombination wild toxin, and contrarily, those of E154R decreased obviously.

Objective:To prepare highly purified Staphylococcal enterotoxin B(SEB) by affinity chromatography and test its activities.Methods:Anti-SEB McAb(1D2) purified by precipitation method with caprylic acid was coupled to Sepharose 4B. And then the SEB was isolated using an affinity chromatography column. In addition, we analyzed the superantigen activity and antigen activity of SEB.Results:The purification efficiency of SEB was 60.71% by affinity chromatography. Its purity was higher than those of standard preparation and the SEB purified by ion change chromatography. At the same time, the purified SEB by affinity chromatography possesses favourable activities of superantigen and antigen.Conclusion:McAb affinity chromatography could be used for purification of SEB with high efficiency.

Objective To evaluate the efficacy of integrated traditional Chinese medicine(TCM)and western medicine in treatment of acute exacerbations of chronic obstructive pulmonary disease(AECOPD)combined with respiratory failure,TCM syndromes of spleen-kidney-yang deficiency and phlegm-dampness by comparison between the integrated therapy and simple western therapy in treatment of the disease. Methods 160 patients with AECOPD combined with respiratory failure,spleen-kidney-yang deficiency and phlegm-dampness syndrome in the intensive care units(ICU)of Affiliated Hospital of Chengdu University of TCM and other four hospitals were randomly allocated into two groups in this double-blinded,multicenter,prospective,randomized,controlled trial. In the control group (78 cases),western medicine and placebo were given to the patients,and in the treatment group(82 cases), conventional western medicine plus fei-shuai mistura 25 mL were administered,four times per day,the therapeutic course lasting for 2 weeks in both groups. The all-cause mortality,respiratory failure-cause mortality,improvement of modified Medical Research Council(mMRC)Dyspnea Scale grades,6 minutes walk distance(6MWD),the forced expiratory volume in 1 second/forced vital capacity(FEV1/FVC)were observed in the 28 days after the end of treatment. Results In the comparisons between the control and treatment groups,there were no statistical significant differences in the all-cause mortality〔54.87%(45/82)vs. 64.10%(50/78)〕and the cases of FEV1/FVC(both P>0.05)in the 28 days after the end of treatment;the 28 day respiratory failure-cause mortality was significantly decreased〔19.51%(16/82)vs. 33.33%(26/78),P<0.05〕,the number of patients with mMRC Dyspnea Scale grades (1-2)was obviously increased(22 cases vs. 7 cases,P<0.05),and the number of patients with 6 MWD grades (4-6)was markedly enhanced in the treatment group(21 cases vs. 8 cases,P<0.05). Conclusions The integrated TCM and western medicine has better therapeutic results in improvement of the patients' degree of dyspnea, 6 MWD and respiratory failure mortality than simple treatment with western therapy for treatment of patients with AECOPD combined with respiratory failure, spleen-kidney-yang deficiency and phlegm-dampness syndrome. However,in regard to the effect on pulmonary function and all cause mortality,the integrated therapy for treatment of such patients in short term has no significant effect.

ObjectiveTo investigate the relation of the human cytomegalovirus pp67-mRNA with　the active infection in renal transplant recipients and the role in the direction for antiviral therapy.Methods　Total 128 samples were collected from peripheral blood treated with EDTA of 32 recipients at the 3rd week　or the 7th week after transplantation.IFA was used to detect CMV pp65 antigen,and NASBA was used to　detect pp67 mRNA assay.Compared with CMV-Ag,the recipients were followed up to observe the change of　pp67 in the cases of the active infection and the CMV disease.ResultsCompared with the proportion between two groups,44 samples out of 128 samples showed CMV-Ag pp65 positive,while pp67-mRNA were　positive in 21 samples.8 patients had been found positive CMV-Ag pp65 and pp67-mRNA; 12 recipients　were clinically diagnosed with HCMV disease.In the early stage of the HCMV infection,the sensitivity and　specificity of the CMV-Ag pp65 ( 91.7％,50.0％ ) was higher than that of pp67-mRNA ( 66.7％,95.0％ ).During the periods of symptoms appeared and the late stage of clinical treatment,the specificity　of pp67-mRNA was higher than that of CMV-Ag pp65,while the sensitivity sit on the contrary.And in the　antiviral therapy course,the indicator of antiviral therapy pp67 turned negative more quickly than the CMVAg,the therapy course could be reduced.ConclusionsBoth of CMV-Ag pp65 and of pp67-mRNA have　clinical significance.Testing for CMV-Ag pp65 is suitable for detecting HCMV active infection at the early　stage,which was important for in time anti-virus treatment in the clinical settings.The test for pp67-mRNA　is a quick method to accquire the accurate results; therefore,it can be used as the reference criteria for the　clinical anti-virus treatment.

Objective To investigate the diagnostic value of the immediate early antigen(IE) mRNA by nucleic acid sequence-based amplification(NASBA) in peripheral blood cytomegalovirus (CMV) infection,and to establish and promote the diagnosis method for CMV.Methods Five to seven ml blood was taken from 32 patients at 3 week and 7 week after renal transplantation to detect serum cytomegalovirus antigen and antibody expression by NASBA,Real time-PCR and enzyme-linked immunosorbent assay (ELISA) sensitivity and specificity were compared.Results The results of CMV detection in 32 renal transplanted patients respectively showed that the positive rate of peripheral blood IE-mRNA by NASBA was 45.8％ (15/32) ;The positive rate of HCMV-DNA in blood by Real time-PCR was 45.8％ (15/32).Using ELISA,the positive rate of HCMV-(IgG +IgM) was 37.5％ (14/32).IE-mRNA and HCMV-DNA had higher sensitivity and specificity and lower false positive rate than HCMV-(IgG +IgM).The positive rates of IE-mRNA by NASBA,Real time-PCR and ELISA were 92.8％,71.5％ and 42.8％ respectively in the 14 cases.Conclusion The nucleic acid amplification method (NASBA based sequence) and Real time-PCR are sensitive,rapid diagnosis methods of HCMV infection,with higher sensitivity and specificity and lower false positive rate than traditional ELISA.And NASBA detection of IE-mRNA has good value for auxiliary clinical diagnosis.

Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P＜0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P＜0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P＜0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.

A new method for O-ethyl S-[2-( diisopropylamino) ethyl] methylphosphonothiolate ( VX) , sarin detection and its kinetic analysis based on piezoresistive microcantilever aptasensor was developed, where VX, sarin aptamers were immobilized on the microcantilever surface by biotin-avidin binding system. A linear relationship between the response voltage and the concentration of VX in the range of 2-60μg/L was obtained. The linear regression equation was △Ue=0. 886C-1. 039 (n=5, R=0. 984, p<0. 001) and the detection limit was 2μg/L ( S/N≥3 ) . A linear relationship between the response voltage and the concentration of sarin in the range of 10-60 μg/L was obtained, the linear regression equation was △Ue=0. 716C-2. 304 ( n=5, R=0 . 996 , p<0 . 001 ) and the detection limit was 10 μg/L ( S/N≥3 ) . The sensor showed no response for O-butyl methylphosphonochloridate, a structural analog of VX and sarin, which indicated high specificity and good anti-interference ability. On this basis, a reaction kinetic model based on receptor-ligand binding and the relationship with output voltage change was established. Response voltage (△Ue ) and response time( t0 ) were obtained from the fitting equation on different concentrations of VX, sarin fitted well with the measured values.

ObjectiveTo investigate the effects of nasal continuous positive airway pressure (nCPAP) and intubation in very low birth weight preterm infants. Methods One hundred and twenty-three very low birth weight preterm infants with respiratory distress within 60 minutes after birth were randomly assigned to nCPAP (n=63) or intubation group (n=60).Outcomes at 7,28 days and 36 corrected gestational weeks were assessed with x2 or t-test. ResultsThere were no significant difference in fatality rate and incidence of bronchopulmonary dysplasia between nCPAP group and intubation group [7.9％ (5/63) vs 6.6％(4/60),4.8％(3/63) vs 3.3％(2/60),x2 =0.07and 0.16,P＞0.05].In nCPAP group,the use of pulmonary sulfactant was 27.0％ (17/63),lower than that (83.3 ％,50/60) in intubation group (x2 =39.34,OR=0.3,90 ％ CI:0.2-0.6,P＜0.05) ;The nCPAP group had fewer ventilation support in 28 days [17.5％ (11/63) vs 25.0％ (15/60),OR=0.7,90％ CI:0.4-1.4] and 36 weeks [6.3％ (4/63) vs 8.3％ (5/60),OR=0.8,90％ CI:0.2-2.4] than those in intubation group but without statistical difference (x2=1.05 and 0.01,P＞0.05,respectively).The incidence of air leak in nCPAP group were lower than intubation group [11.1％ (7/63) vs 33.3％ (20/60),x2 =8.86,OR=0.3,90％00 CI:0.2-0.7,P＜0.05].There was no significant difference for other complications between two groups. ConclusionsIn very low birth weight preterm infants,early nCPAP dose not significantly reduce the fatality rate and the incidence of bronchopulmonary dysplasia as compared with intubation ventilation,but shorten the time of ventilation and lower the incidence of air leak.

Objective:To investigate the clinical efficacy of partial excision of nail matrix combined with phenolic ablation in the treatment of ingrowing toenail.Methods:115 patients(148 toenails) with ingrowing toenail treated in the Central hospital of Wuhan from October 2004 to May 2013 were selected for this study.The patients were divided into two groups,53 patients(67 toenails) admitted from October 2004 to December 2007 were considered as the observation group and treated with Partial excision of nail matrix,62 patients(81 toenails) admitted from January 2008 to May 2013 were considered as the control group and treated with Partial excision of nail matrix and phenolic ablation.The bleeding time,pain relief time,healing time,recurrences after one year and satisfaction rate of two groups were compared.Results:The wounds of 148 toenails were healing.The Bleeding time,pain relief time,recurrences after one year in the control group (1.85:±:0.42days,13.25± 2.17hours) were lower than the observation group (2.69± 0.53 days,21.54± 2.56hours),and healing time in the control group (11.32± 2.37days) were longer than the observation group (8.93± 2.06 days)(P＜0.05),the recurrence rate and overall satisfaction rate of observation group and control group were 6.15％,12.5％ and 97.06％,91.07％.Conclusion:Partial excision of nail matrix combined with phenolic ablation was more effective in the treatment of ingrowing toenail than surgical excisional techniques.

Objective To determine the risk factors for imipenem-resistant Pseudomonas aeruginosa (IRPA) infections in neonatal intensive care unit (NICU).Methods One hundred and eighty-eight Pseudomonas aeruginosa infected children (confirmed by pathogenic examination) in the NICU of Maternal and Child Health Hospital of Guangdong from January 1,2008 to December 31,2011,were chosen as the objects of study,and were divided into two groups.The first group included 73 children that had been isolated with IRPA strains(IRPA group),and the second group included 115 children that only had imipenem-sensitive Pseudo monas aeruginosa (ISPA) strains (ISPA group).Chi-square test or t-test was applied.The risk factors were investigated by univariate or multivariate Logistic regression analysis.Results Data from univariate analysis showed that the gestational age and birth weight of neonates were lower than those in ISPA group [(34.0±3.5) weeks vs (35.6±2.8)weeks,t=3.413,P＜0.01; (1848.1±276.4) g vs (2110.7±345.6) g,t=5.472,P＜0.01].There were more neonates with gestational age ≤ 32 weeks [67.1％ (49/73) vs 45.2 ％ (52/115)],birth weight ＜1500 g [73.9％ (54/73) vs 33.0％(38/115)],small for gestational age [68.5％ (50/73) vs 29.6％ (34/115)],receiving imipenem [72.6％ (53/73) vs 27.0％ (31/115)] or the third generation cephalosporins [65.8％ (48/73) vs 33.0％ (38/115)] two weeks before the isolation of Pseudomonas aeruginosa,and mechanical ventilation [78.1％ (57/73) vs 61.7 ％ (71/115)],deep vein catheterization [83.6％(61/73) vs 65.2％(75/115)] in the IRPA group than in the ISPA group (all P＜0.05 or 0.01).The multivariate Logistic regression analysis revealed that imipenem treatment within two weeks before the isolation of Pseudomonas aeruginosa (OR=6.409; 95％ CI:1.926-21.333,P =0.002) was an independent risk factor.Conclusions IRPA infection in NICU hospitalized infants is strongly related to their gestational age and birth weight.History of imipenem administration could be an independent risk factor.