Objective　To obtain staphylococcal enterotoxin B (SEB) mutant with normal antigenicity but low toxicity. Methods　Using PCR technique, normal SEB (SEB-N) gene which was amplified from S. aureus S6B. SEB mutant gene (SEB-M) was prepared from the same strain, but one nucleotide in SEB gene was changed from asparagine (N23) to serine (S23). SEB-N and SEB-M were cloned into procaryotic expression vector pTrc99A then and transferred into E. coli JM109. SEB-N and SEB-M which were cloned into plasmid were sequenced directly by dideoxynucleotide method. The crude expressed proteins were identified by double agar immunodiffusion. The level of IL-2 in supernatants of mouse splenocytes stimulated by crude expressed proteins was determined by ELISA. Results　SEB-N and SEB-M were obtained through PCR. The sequence of SEB-N was changed with non site-directed mutagenesis, threonine at the residue 150 of SEB-N was replaced with alanine (ACT→GCT, T150A). As being expected, at the residue 23 of SEB-M, serine substituted for asparagine (AAT→AGT, N23S) with site-directed mutagenesis. Double agar immunodiffusion showed obvious precipitin line with anti-SEB by both crude SEB-N and SEB-M mutant proteins could produce, but not by non-recombinant strain. ELISA demonstrated that the level of IL-2 in supernatant of mouse splenocytes stimulated by natural SEB protein (containing equal amount of JM109P crude protein) was 40 times as much as that stimulated by SEB-M and 12.5 times as much as that stimulated by SEB-N. Conclusions　We obtained two recombinant strains which produced T150A and N23S mutant SEB protein. The mutant proteins showed binding ability to anti-SEB as the normal protein. However, their biological activity as superantigen decreased sharply. We consider that it is promising for further study of molecular adjuvant or superantigen vaccine.

Objective To analyze risk factors for acute kidney injury after aortic arch operation under deep hypothermic circulatory arrest.Methods Between January 2005 and June 2011,549 cases aortic arch replacement under deep hypothermic circulatory arrest were retrospectively analyzed.According to the occurrence of acute kidney injury they were divided into two groups.Univariate and multivariate analysis (multiple logistic regression) were used to identify the risk factors.Results AKI occurred in 102 cases (18.6％) and 27 cases(4.9％) had dialysis.Multiple logistic regression showed that body mess Index(OR =1.072,95％ CI:1.006-1.141,P =0.031),serum creatinin (OR =1.011,95％ CI:1.006-1.017,P =0.000),cardiopulmonary bypass time(OR =1.006,95 ％ CI:1.002-1.009,P =0.005) and the peak intraoperative glucose level (OR =1.007,95 ％ CI:1.002-1.011,P =0.003) were independent risk factors for AKI.Conclusion The higher BMI,serum creatinin level maybe indicate the occurrence of AKI,and AKI maybe can be reduced by controlling CPB time and intraoperative hyperglycemia.

ObjectiveTo investigate the clinical features and pregnant outcomes of the pregnant women with congenital adrenal hyperplasia (CAH) 21-hydroxylase deficiency (21-OHD).MethodsThe clinical features,therapies,pregnant outcomes of the pregnant women with 21-OHD were retrospectively reviewed in Peking Union Medical College Hospital,from January 2005 to April 2011.ResultsThere were 8 pregnant womenwith 21-OHD including 5siinplevirilizing patientsand 3nonclassical 21-OHD women.Eightpatientswereacceptedprogestationalandprenatalcontinuallowerglucocorticoid treatment.During the gestational period,the dosage of glucocorticoid was adjusted in one pregnancy.The serum level of 17-alpha hydroxyprogesterone (17-OHP) were elevated after pregnancy [ (70 ± 38 ) versus (24 ±23) nmol/L,P ＜ 0.05].The fertility and offspring rate of 8 patients was 8/12,the fertility and offspring rate of patients who started treatment at preadolescence was significantly increased (4/5 versus 4/7).Four patients were accepted genital reconstructive surgery ( clitorectomy,clitoroplasty,vulvoplasty) before pregnancy.The incidence of GDM was 1/8.All patients selected caesarean at from 37 +6 gestation weeks to 39+6 gestation weeks.The average newborn birth weight was (3210 ± 447 ) g,and height was (48 ±2) cm of 8 neonates,none of them was CAH.Conclusions Medical and surgical therapy provides satisfactory fertility and pregnancy outcomes for women with 21-OHD.It is safe to pregnant women with 21-OHD and their fetus in continual lower glucocorticoid treatment.The dosage of glucocorticoid should be carefully adjusted during the pregnancy individually according to serum level of 17-OHP.

Objective To investigate the effects of different therapeutic strategies on recurrence of postoperative Crohn's disease (CD) patients.Methods From September 2009 to September 2014,85 CD patients with intestinal resection were enrolled.The clinical features and maintenance therapeutic medication were retrospectively analyzed.The patients were divided into non-treatment group (induding continuously or cumulatively taking medicine less than three months),5-aminosalicylic acid (5-ASA) group and immunosuppressant agents group (including azathioprine,methotrexate and thalidomide).Kaplan-Meier method was performed to compare the recurrence rate in postoperative CD with different therapeutic medication and the risk factors of postoperative recurrence were also analyzed.Results Among 85 CD patients,there were 32,21 and 32 patients in non-treatment group,5-ASA group and immunosuppressive agents group,respectively.After surgery,the one year accumulated clinical recurrence rate of immunosuppressant agents group was 12.5％ (4/32),which was significantly lower than that of non-treatment group (56.3％,18/32) and 5-ASA group (38.1％,8/21),and the differences were statistically significant (x2 =12.250,P＜0.01;x2 =4.102,P =0.043).After surgery,the two years accumulated clinical recurrence rate of immunosuppressant agents group was 12.9 ％ (4/31),which was significantly lower than that of non-treatment group (75.9％,22/29) and 5-ASA group (47.6％,10/21),and the differences were statisitcally significant (x2 =17.840,P＜0.01;x2 =6.597,P=0.010).After operation,the one year accumulated endoscopic recurrences rates of non-treatment group,5-ASA group and immunosuppressant agents group were 39.1％ (9/23),5/16 and 34.6％ (9/26),respectively;while the two year accumulated endoscopic recurrence rates were 59.1％ (13/22),6/16 and 44.0％ (11/25),respectively.However,there was no statistically significant difference among the groups (all P＞0.05).Penetrating lesion was an risk factor of postoperative clinical recurrence in CD patients (x2 =4.963,P=0.026,oddsratio (OR) =2.221,95 ％ confidence interval (CI) 1.121 to 5.775).Conclusions Immunosuppressive agents rather than 5-ASA have remarkable effects in preventing postoperative clinical recurrence in CD patients.Postoperative clinical recurrence is more likely to happen in patients with penetrating lesions.

In this paper the application of DNA fingerprints to evaluate the efficiency of DNA extraction was studied DNA mixtures of landfill leachate and active sludg e were extracted using three different methods and the DNA extraction was evalua ted by ARDRA and RISA fingerprints The results showed that RISA is a effecti ve way to evaluate DNA extraction

Objective:To evaluate the clinical efficacy and safety ofHJ-RY-1 therapeutic pad ofsoft stone in treating osteoarthritis.Methods:Patients with osteoarthritis with syndrome ofqi stagnancy and blood stasis were enrolled according to the diagnostic criteria and the evaluation standard ofcurative effect in Instruction Principle ofClinical Research ofNew Chinese Medicines, and were randomly divided into experimental group(treating by HJ-RY-1 therapeutic pad ofsoft stone) and the control group(treating by the paster offar infrared bioactive ceramics).Put them respectively on the afflicted part for more than 12 hours a day, and the treatment course ofboth groups lasted 14 days.The main turnover was determined by the improvement of6 clinical parameter indices and the incidence rate ofwhole curative effect and side effect after a treatment of7 days and 14 days.Results:72 patients were randomly equally divided into two groups, and received predetermined interveneing therapy.There were similar baseline characteristics and comparabilities between them.Totally 68 patients finished 2 weeks follow-up observation and 4 cases were missed(33/36 in experimental group;35/36 in the control group).1 weeks later, there were no significant difference in clinical situation and overall curative effect between two groups.But after two weeks interveneing therapy, there were significant difference on activity pain(t-value was 3.3104), the joint movement range(t-value was 3.1596), self-sensation(t-value was 2.5521) and overall curative effect(t-value was 3.5694) between them(P

Objective To analyze risk factors for cerebral neurological injury after operation of acute Stanford type A aortic dissection.Methods Between January 2005 and December 2011,329 cases of acute Stanford type A aortic dissection patients underwent aortic arch replacement were retrospectively analyzed.Univariate and multivariate analysis(multiple logistic regression) were used to identify the risk factors for postoperative cerebral neurological injury including permanent neurological dysfunction (PND) and temporary neurological dysfunction (TND).Results Cerebral neurological injury occurred in 77 cases (23.4％),PND 11 cases(3.3％) and TND 66 cases(20.1％).Multiple logistic regression showed that age(OR =1.087,95％ CI 1.013-1.166,P =0.020) and stroke history (OR =10.383,95 ％ CI 1.596-67.534,P =0.014)were independent risk factors for PND,serum creatinin (OR =1.013,95 ％ CI 1.004-1.023,P =0.006),WBC (OR =1.199,95 ％ CI 1.087-1.324,P =0.000) and peak intraoperative glucose level (OR =1.011,95％ CI 1.004-1.018,P =0.003) were independent risk factors for TND.Conclusion The older age or stroke history indicate the occurrence of PND,Whereas the higher WBC lever preoperative or higher intraoperative glucose level indicate the occurrence of TND.The incidence of TND probably will be reduced by controlling intraoperative hyperglycemia actively.

Objective To analyze risk factors for in-hospital death of aortic arch replacement under deep hypothermic circulatory arrest and antegrade cerebral perfusion.Methods Between January 2005 and December 2011,626 cases aortic arch replacement under deep hypothermic circulatory arrest and antegrade cerebral perfusion were retrospectively analyzed.The cause of death were recorded and univariate and multivariate analysis (multiple logistic regression) were used to identify the risk factors.Results In-hospital death occurred in 29 cases (4.6％) and in them 2 cases because of aortic dissection rupture,5 cases severe neurological injury,11 cases low cardiac output syndrome and 11 cases multiple organ failure.Multiple logistic regression showed that stroke history(OR =6.703,95％ CI:1.664-27.000,P ＝ 0.007),preoperative hemodynamic instability(OR =6.441,95％ CI:1.213-34.212,P =0.029),eardiopulmonary bypass time(OR =1.008,95％ CI:1.002-1.014,P =0.007) and CABG(OP =4.525,95％ CI:1.542-13.279,P =0.006) were independent risk factors for in-hospital death.Conclusion Stroke history,preoperative hemodvnamic instability and coronary involvement indicate high risk in operation,and mortality maybe can be reduced by controlling CPB time.

Objective To investigate the impact of acute kidney injury ( AKI ) on early and long-term outcome using RIFLEcriteria in patients after acute type A dissection surgery.Methods Between January 2006 and June 2011, 286 cases a-cute type A dissection patients underwent deep hypothermic circulatory arrest surgery were retrospectively analyzed .Using RIFLEcriteria to classfy AKI to risk, injury and failure stages and comparing 30-day mortality, dialysis, ventilation time, ICU stay, hospital stay and cost, and three-year mortality.Multivariate analyses were performed to identify predictors of failure stage.Results AKI was detected in 160 patients(55.9%), and risk 85 cases(29.7%), injury 39 cases(13.6%), failure 36 cases(12.6%).Dialysis rate, ventilation time, ICU stay, cost, and 30-day mortality increased with worsening severity of renal injury, and these was significant increased in failure stage than other groups .There was no association between severity of renal injury and 3-year mortality.Multiple logistic regression showed that malperfusion syndrome ( OR =3.499, 95%CI:1.019-12.013, P=0.047) and WBC(OR=1.121, 95%CI:1.017 -1.237, P=0.022) were independent predictors of renal failure.Conclusion Postoperative mild and morderate acute kidney injury were common in acute type A dissection pa-tients.The severe acute kidney injury had association with poor early outcome .

AIM: To evaluate the ability of spectral domain optic coherence tomography ( SD-OCT ) parameters to detect progressive structural damage in primary open angle glaucoma ( POAG) by contrasting with visual field. ·METHODS: Retrospectively we evaluated 48 subjects (48 eyes ) of POAG, followed up 14 to 62mo. The parameters of SD-OCT and visual field were obtained. The correlation between the change of visual field measurements and OCT measurements were analyzed. Visual field progression was defined as reproducible drop of at least 2dB of mean deviation (MD) from the baseline visit. ROC curve was made to evaluated the ability of OCT parameters in detect progression of POAG. ·RESULTS: The 25 eyes were classified in progression group and 23 eyes were in non-progression group. No significant correlation was seen between OCT parameters and visual field in non-progression group. In progression group, OCT parameters which were significantly correlated with MD reduce were rim volume ( r=-0. 5997, P=0. 0007), C/D vertical ratio (r=-0. 6309, P=0. 0003), RNFL(r= 0. 4201, P= 0. 0260), and GCC(r= 0. 7080,P<0. 01). ROC curves showed the GCC reflected the reduce of MD accurately (P=0. 013). ·CONCLUSION: Part parameters of SD-OCT can reflect the progression of POAG accurately and provided a new method to detect the damage of visual function of POAG.