Objective To obtain sufficient recombinant multiepitope peptide of ESAT-6.Methods DNA encoding ESAT-6 peptide was amplified by PCR using a pair of primers which were designed in accordance with the reported ESAT-6 gene sequence.The ESAT-6P gene was inserted into the prokaryotic expression vector pET-30a with the N-terminal 6His-tag.The recombinant peptide vector pET30a-ESAT-6P was transformed into E.coli BL21(DE3) via chemical transformation.The positive clone was screened by means of PCR.The target peptide was expressed in E.coli after induction with IPTG and analyzed by SDS-PAGE.The immunogenicity of the peptide was evaluated by dot immunobinding assay.Results The target peptide was successfully expressed and purified.The solubility analysis showed that the recombinant peptide existed as inclusion body in the E.coli.DIBA indicated that the ESAT-6P was specifically reactive to sera from TB patients.Conclusions The recombinant multiepitope peptide of ESAT-6 engineering bacteria has been obtained,which will be quite helpful to develop novel specific diagnostic reagents and effective vaccine.

This paper presents a new method of separating the junction between the epidermis and the dermis with enzyme,and then,observing the small wound of the papillary of the dermis by means of SEM.The result showed that this new method would be advantageous to the identification of small stab wounds and providing some concrete identify targets.

Objective To compare the changes of serum levels of heme oxygenase‐1(HO‐1) ,C‐reactive protein(CRP) and Chitinase‐3‐1ike protein 1(YKL‐40) in patients with rectal cancer after laparoscopic‐assisted radical operation versus open radical surgery .Methods According the integration and elimination standard ,60 cases of patients with rectal cancer were selected and di‐vided into laparoscopic group (30 cases) and open group (30 cases) ,the concentrations of HO‐1 ,YKL‐40 and CRP in peripheral blood of patients were detected on the first and third postoperative day and preoperative day(P<0 .05) ,and were compared between the two groups .Results The serum levels of HO‐1 ,YKL‐40 and CRP in the laparoscopic group were significantly lower than those in the open group on the first and third postoperative day(P<0 .05) .The serum levels of HO‐1 ,YKL‐40 and CRP were significant‐ly increased after surgery ,and serum levels of HO‐1 and CRP on third postoperative day were significantly increased compared with that on the first postoperative day in the two groups after surgery ,while serum level of YKL‐40 on the third postoperative day was significantly lower than that on the first postoperative day in the two groups after surgery .Conclusion The stress reaction is com‐paratively mild in patients with rectal cancer after laparoscopic assisted radical operation ,and the postoperative period may be shor‐ter in patients treated with laparoscopic assisted radical operation than those treated with open radical surgery .

Objective To investigate the changes of the electrocardiograms (ECG) and the cardiac markers in patients with acute insular infarction,and analyze the relationship between them and the prognosis.Methods A total of 202 patients with acute middle cerebral artery territory infarction (patients group) and 150 control subjects (control group) was selected in this study.Patients included insular infarction (insular infarction group,136 cases),non-insular infarction (non-insular infarction group,66 cases),left-side insular infarction(71 cases) and right-side insular infarction(65 cases).ECG recordings and plasma cardiac troponin I (cTnI),creatine kinase-MB (CK-MB) were measured and compared.Death in 6 months was followed-up.Results There was significant difference in the incidence of abnormal changes of ECG and plasma cTnI,CK-MB increasing between patients group and control group (P ＜0.01).The incidence of abnormal changes of ECG and fatality rate were higher in insular infarction group than those in non-insular infarction group [80.88％(110/136) vs.46.97％(31/66) and 11.76％ (16/136) vs.3.03％ (2/66),P ＜ 0.05 or ＜ 0.01].The incidences of ectopy and prolonged QT were higher in right-side insular infarction patients than those in left-side insular infarction patients [44.62％(29/65) vs.11.27％ (8/71),P ＜0.01 ; 55.38％ (36/65) vs.35.21％ (25/71),P ＜ 0.05].The incidences of sinus bradycardia and ST segment deviation were higher in left-side insular infarction patients than those in right-side insular infarction patients [22.54％(16/71) vs.7.69％(5/65),P ＜ 0.05 ;47.89％(34/71) vs.13.85％ (9/65),P ＜ 0.05].The increased rates plasma cTnI and CK-MB level were mainly seen in insular infarction [insular infarction group:47.79％ (65/136),34.56％ (47/136); non-insular infarction group:4.55％ (3/66),1.52％ (1/66),P ＜ 0.01].The incidence of plasma cTnI increasing in right-side insular infarction patients was higher than that in left-side insular infarction patients [67.69％(44/65) vs.29.58％(21/71),P＜ 0.05].There was no significant difference in the incidence of plasma CK-MB increasing between left-side insular infarction patients and right-side insular infarction patients(P ＞ 0.05).The fatality rates in plasma cTnI,CK-MB increasing patients were higher than those in normal plasma cTnI,CK-MB patients [16.18％ (11/68) vs.5.22％ (7/134),P ＜0.05;29.17％ (14/48) vs.2.60％ (4/154),P ＜0.01].Conclusions The effects of acute hemispheric cerebral infarction on heart are mainly associated with destruction of insula.Patients with insular infarction have more abnormal changes of cardiac markers and ECG,which is correlated with poor prognosis.

Objective To analyze the pathogen distribution and drug resistance of candida isolated from children with blood infections in our hospital,and to provide reference for clinical effective prevention and treatment.Methods The blood specimens of pediatric patients were collected between January 2009 and December 2015,and were cultured using BacT/ALERT 3D and BD9140 instruments.The candida were separated with Sobaurandps agar culture medium,and identified with chromogenic medium,API 20CAUX test strips or VITEK-2 compact YST card.The minimal inhibitory concentration of 5 drugs were determined by ATB FUNGUS 3 system.Results In 176 cases,92 strains (52.3％) were from neonatal ward,and 46 strains (26.1％) were from PICU.In newborn group,85 strains were isolated from premature,which contained the low and very low birth weight infants (37 strains),pneumonia(20 strains),neonatal respiratory distress syndrome(9 strains).In PICU,the strains were commonly isolated from children with severe infection.Among 176 strains of candida,71 strains (40.3％) were C.albicans,62 strains (35.2％) were C.parapsilosis,16 strains(9.1％) were C.glabrata,9 strains(5.1％) were C.tropicalis,and 18 strains(10.2％) belonged to other candida.Conclusion Candida blood infections can happen at all age of chlidren.The most common strains detected from blood were C.albicans,followed by C.parapsilosis.Most of these strains are susceptible to antifungal drugs,such as fluconazole,except C.glabrata.The sensitive rates to commonly used antifungal drug are more than 93％.The selection of antifungal drugs should be based on the species of strains.

Objective To evaluate the changes in the expression of DJ-1 protein during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Fifty male Sprague-Dawley rats,weighing 220-280 g,were used in this study.Type 1 diabetes mellitus was induced by intraperitoneal streptozotocin 65 mg/kg and confirmed by fasting blood glucose ＞ 16.7 mmol/L.Forty animals with type 1 diabetes mellitus were randomly divided into 3 groups:diabetes group (group D,n =10),diabetic sham operation group (group DS,n =15) and diabetic I/R group (group DIR,n =15).Another 10 non-diabetic rats in which citrate buffer 6 ml/kg was injected intraperitoneally were served as control group (group C).Myocardial I/R was produced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in group I/R.At 120 min of reperfusion,5 rats were sacrificed and myocardial specimens were c(on)tained for determination of infarct size in groups DS and DIR,and 10 rats were sacrificed and myocardial specimens were obtained for microscopic examination and for determination of cell apoptosis,malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of DJ-1 and phosphatase and tensin homologue (PTEN) protein.Apoptotic index (AI) was calculated.Linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein was analyzed.Results Compared with group DS,the myocardial infract size was significantly increased in group DIR (P ＜ 0.05).Compared with group C,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in groups D,DS and DIR (P ＜ 0.05).Compared with groups D and DS,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in group DIR (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups D and DS (P ＞ 0.05).There was linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein and the correlation coefficients (r) were-0.734,0.593,-0.818,and-0.812 in turn.Conclusion Down-regulation of DJ-1 protein expression is involved in myocardial I/R injury in diabetic rats via decreasing anti-oxidative stress responses and upregulating PTEN protein expression.

Objective To evaluate the outcome of patients using intra-aortic balloon pump IABP undergoing coronary artery bypass grafting and to assess the results.Methods From December 2001 to December 2011,eighty-two consecutive patients,who used IABP undergoing coronary artery bypass grafting were studied.The out-come of early complication,mortality,LVEF were analyzed.Results Fourteen patients died.Forty-three patients,who used IABP,no serious complications (malignant arrhythmia,renal insufficience),preoperative LVEF was (46.0 ± 1.8) ％,LVEDD was (66.0 ± 4.1) mm,early postoperative LVEF was (50.0 ± 2.7) ％,LVEDD was (53.0 ± 2.8) mm (P ＜ 0.05).Conclusion Use of IABP decreased early mortality,and postopertive respiratory tract infections,renal insufficiency were the main complication.Positive use of IABP postoperatively can decrease early mortality.

Objective To investigate the role of p38 mitogan-activated protein kinase signal transduction pathway in lipopolysaccha-ricle-indaced acute lung injury (ALI) in rats. Methods 48 wistar rats were randomly divided into 3 groups, saline control group (group A), lipopolysaccharide (LPS) group(group B) and SB203580 group(group C). Models of lipepelysaccharide-imluced ALI were used to oh-serve the expression of p38MAPK in rat lung, protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF) , pulmonary MDA content and the activities of serum NO. After LPS dripping for 6 hours, arterial blood was drawn for analysis and lung tissue was detec-ted. Results Compared with those in group A, expression of p38MAPK were markedly increased in group B and C (P<0.01). But in group C, expression of P38MAPK was significantly lower than that in group B (P<0.05). The protein content, the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), content of pulmonary MDA and the activities of serum NO in group B, C were significantly higher than those in group A (P<0.01). There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in group B and C (P<0.01). Compared with those in group B, these indexes of lung injury were significantly lower while the level of arterial bicar-bonate and partial pressure of oxygen was increased in group C(P <0. 05 orP <0. 01). Under light microscope, the pathologic changes in-duced by LPS were significantly attenuated by SB2035g0. Conclusion The activation of P38MAPK play an important role in the mechanism of lipopolysaccharide-induced ALI.

Objective To investigate the effect of radix paeoniae rubra (RPR) on expressions of p38MAPK, NF-κB and iNOS in ltpopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats with ALI were divided randomly into five groups: saline control group (Group A) , LPS group (Group B), RPR for treatment group (Group C), RPR for prevention group (Group D) and SB203580 group (Group E). The effects of RPR on protein content and the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), contents of malondidehyde (MDA) and serum NO in lung tissue were observed. Arterial blood was collected for blood gas analysis. Expressions of p38MAPK, NF-κB and iNOS in rat lung tissues were detected. Results Compared with Group A, expressions of p38MAPK, NF-κB and iNOS were significantly increased (P <0.01 orP< 0.05) , the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly higher (P<0.01 or P<0.05) in Groups B, C, D and E. There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Groups B, C, D and E (P<0.01 or P<0.05). Compared with Group B, the expressions of p38MAPK, NF-κB and iNOS were significantly lower, and the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly decreased, while the levels of arterial bicarbonate and partial pressure of oxygen were significantly higher in Groups C, D and E (P<0.05 or P<0.01). Compared with Group B, the lunginjury of Groups C, D and E was significantly alleviated, while there was no statistical difference among Groups C, D and E in the indices of lung injury. Conclusion Protective effect of RPR on LPS-induced ALI is closely related to the inhibited expressions of p38MAPK, NF-κB and iNOS.