To prokaryotic express prokaryotically and to purify the efaA protein from Enterococus faecalis so as to provide the basis for the further study on the pathogenesis and clinical sero-diagnosis of endocarditis caused by E.faecalis, efaA gene of E.faecalis was amplified by PCR, the PCR-amplified product was digested with restriction enzymes and cloned into prokaryotic vector pET32a to construct the recombinant plasmid pET30a/efaA. This recombinant plasmid was confirmed by double enzyme digestion with BamhI and Xhol and then subjected to sequencing, and transformed to E.coli BL21 (DE3). Expression of the fusion protein was induced by IPTG, and analyzed by SDS-PAGE and Western blotting. The recombinant fusion protein was purified by His-binding affinity chromatography.It was shown that efaA gene of 943 bp in size was amplified from Enterococus faecalis and the recombinant plasmid pET30a,/ efaA was successfully constructed and expressed in E.coli BL21. The purified product was found to be 34 kDa in molecular weight as demonstrated by SDS-PAGE and Western blotting. It is evident that the efaA protein of E.faecalis can be successfully expressed and purified.

10.3969/j.issn.2095-4344.2013.25.024

Objective To study prognosis of patients with fulminant hepatitis after plasma ex- change treatment using model for end-stage liver disease(MELD)scoring system.Methods 160 pa- tients were randomly divided into plasma exchange group and control group,and MELD score was calculated according to the original formula for each patient.The efficacy of plasma exchange was as- sessed by mortality and improvement in biochemical parameters and MELD score.Results The levels of total bilirubin(TBIL),INR and MELD score of patients whose MELD scores were between 30 and 40[TBIL,(379.4?40.4)?mol/L; INR,2.5?0.2; MELD,30.8?3.8]were lower than before PE treatment[TBIL,(509.7?64.6)?mol/L;INR,3.5?0.3;MELD,37.3?3.5].The levels of TBIL and INR and MELD score of patients whose MELD scores were higher than 40 [TBIL,(595.6?61.5)?mol/L;INR,3.8?0.4;MELD,39.8?3.5]were lower than before PE treatmem [TBIL, (650.4?66.3)?mol/L;INR,4.4?0.6;MELD,45.2?4.2].The mortality of patients in PE group with MELD score from 30 and 40 was 50.0%,while it was 86.7% in control group,showing significant differ- ence between PE group and control group(P＜0.01).The mortality of patients with MELD scores higher than 40 was 91.2% in PE group and 100% in control group,showing no significant difference between these two groups(P＞0.05).Conclusions Plasma exchange treatment can decrease the serum TBIL level, INR and MELD score of patients with fulminant hepatitis and improve liver function.Compared with the control group,plasma exchange can significantly decrease the mortality of patients in PE group with MELD score from 30 to 40,but no effect on patients with MELD score higher than 40.

Objective To evaluate the clinical safety and efficacy of HIFU treatment of hepatic metastases from colorectal cancer.Methods 20 tumor lesions in 14 patients with hepatic metastases from colonectal cancer were treated with HIFU therapy.The vital signs,main organs'functions,examination of MRI and complications were observed.Results Vital signs of all the 14 patients were stable.The function of liver and kidney were normal and life quality were improved with elevation of Karnofsky in 5 patients.On MRI examination,the MRI signals of treating area of 20 tumor lesion in 14 patients decreased remarkably with disappearance of blood supply and a thin layer of edematous intensified zone was detected on the margin of lesion.6 patients were found to have cord-like burning scar on skin.Conclusion HIFU treatment is safe and effective in treatment of hepatic metastases from colorectal cancer.

Bronchi ; Diagnostic Errors ; Female ; Foreign Bodies ; Humans ; Middle Aged

Objective: To observe the changes of heart rate and the contents of angiotensin Ⅱ(AngⅡ) in blood plasma in ventricular tachycardia(VT) rats with electro-acupuncture(EA) on ‘daling’(PC7). Methods: 40 SD rats were randomly divided into normal, model, treatment and control groups with 10 cases in each. VT model was duplicated by inject CsCl in femoral vein. To observe the rats’ electrocardiogram (ECG) and record their heart rates. EA was applied to‘daling’ (PC7) on treatment group and applied to‘Taiyuan’ (LU9) on control group for 5 minutes. Then, we detected the contents of AngⅡ in the rats’blood plasma respectively. Results: The heart rate and contents of AngⅡin rat increased obviously in model group were (547?30) time/min and (353.21?49.12)pg/mL). They restored to the normal state after EA ‘daling’ (PC7) are(474?25)time/min and(268.44?47.49)pg/ mL.But the effect was not obvious in ‘Taiyuan’ (LU9). Conclusion VT rat heart can be prompted to restore to the normal state after EA ‘daling’(PC7); AngⅡ played an important role in VT.

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Breast ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Female ; Fibrocystic Breast Disease ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Middle Aged ; S100 Proteins ; metabolism

The strain F12-11-1-2 was isolated from the East China Sea,which had antimitosis activity using Pyricularia oryzae mode.Ac- cording to phenotypical study,salt-aggregation test and 16S rDNA sequence analysis,the strain F12-11-1-2 has been identified to be Bacillus subtilis.

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.