A Staphylococcus aureus strain, designated zfb, was isolated from a clinical bovine mastitis case of a dairy cow. Staphylococcus aureus zfb can have resistance to methicillin and no lipase contrast by ATCC 25923. The production of the capsule was assessed by the diffuse colonial morphology in serumsoft agar. A mouse infection model was used to determine the LD50 and the invasiveness of SA zfb. The LD50 of SA 25923 to experimental mice was 10-2.5/mL, and the LD50 of SA zfb to experimental mice was 10-4.33/mL. The purpose to detect characteristics of SA zfb makes it an interesting candidate for the preparation and assay of an avirulent mutants against staphylococcal infections and further investigate on pathogenic mechanism.

Objective:? -Hemolysin of Staphylococcus aureus,which was expressed in E.coli BL21(DE3) with recombinant pET32a+-?-HL plasmid,was purified with gel filtration chromatography(GFC),and then the engineered subunit vaccine was developed. The immunity effectiveness of this vaccine was evaluated on mouse models.Method:The purified fusion protein was analyzed in SDS-PAGE,and subjected to the evaluation of its median hemolytic dose potency(HD50) was finally analyzed with rabbit erythrocyte. Protein concentration was determined by the method of Bradford. Antibody titers were evaluated on ELISA,and then challenged to gain the immunity protect index.Results:There is an expected protein band with molecular mass of 53kDa in SDS-PAGE,and the concentration is 0.1278mg/ml. The hemolysis activity is 8012.5 HU/mg. There are specific antibodies acquired in blood-serum from mice after vaccined and the antibody titers rising until it has arrive the max,then following down.Conclusion:The purified fusion protein has good fineness and hemolysis activity,the antibody titers initiated by the protein vaccine go with regulation and the immunoprotection is satisfied.

BACKGROUND: Acute renal insufficiency (ARI) usually occurred following liver transplantation due to the surgical trauma and the application of immunosuppressant, which lack of unified diagnostic criteria. OBJECTIVE: To investigate the experience of diagnosis and treatment of ARI following liver transplantation.DESIGN, TIME AND SETTING: The experiment was performed at the 458 Hospital of Chinese PLA from January 2004 to December 2006.PARTICIPANTS: A total of 37 cases received liver transplantation, including 35 males and 2 females, aged 37-67 years, mean aged (48.5±8.9) years. All cases were divided into the liver cancer group (n=16) and liver cirrhosis group (n=21). The liver cirrhosis group included 16 cases with posthepatitic type B cirrhosis, 4 with posthepatitic type C cirrhosis, and 1 with alcoholic cirrhosis. All these cases were in decompensation stage. The final diagnosis was performed by pathological examination. METHODS: The removal of kidney and construction of blood outflow tract was achieved by modified piggy-back liver transplantation. The arterial blood gas analysis, blood routine examination, renal function and liver function were examined more than twice per day. The cephalosporins, Fluconazole and ganciclovir or vancomycin were used for 5-7 days to prevent infections.MAIN OUTCOME MEASURES: The incidence rate of acute ARI, clinical features and outcomes of patients were observed.RESULTS: ARI developed in 19 patients with liver transplantation, 5 patients died, 14 patients recovered in 2-3 weeks. The incidence of ARI following liver transplantation was associated with infection, bleeding shock, respiratory failure and acute respiratory distress syndrome (P < 0.05). CONCLUSION: The incidence of ARI following liver transplantation was 51.35%, with 26.32% mortality rate. The early diagnosis and treatment are the key steps for increasing successful rate of ARI treatment following liver transplantation.

Objective To investigate the effects of Qishen Erlian Decoction on serum MMP-1 and TIMP-1 levels in thioacetamide (TAA) induced liver fibrosis rats; To discuss its mechanism of action. Methods Liver fibrosis model was created by the TAA gavage method. 120 SD male rats were randomly assigned to control group, model group, colchicine group, Qishen Erlian Decoction low-, medium- and high-dose group (20 in each group). Each medication group was given relevant medicine for gavage. Control group and model group were given the same amount of normal saline for gavage, once a day for 5 weeks. HE staining and Masson trichrome staining were used to observe the pathological changes in liver tissue and liver tissue damage. Biochemistry, radioimmunoassay, and ELISA were used to detect the serum liver function, hepatic fibrosis index, MMP-1 and TIMP-1 levels. Results Compared with the model group, serum ALT, AST, TBIL, γ-GGT, HA, LN, Ⅳ-C and PCⅢ levels, MMP-1 and the ratio of MMP-1/TIMP-1 increased significantly and level of TIMP-1 decreased significantly in Qishen Erlian Decoction groups, with statistical significance (P<0.05, P<0.01). And there is a certain dose-effect relationship, with Qishen Erlian Decoction high-dose group the best effect. Conclusion Qishen Erlian Decoction can improve the liver function and liver fibrosis indexes, regulate the level of MMP-1 and TIMP-1, and prevent the progression of liver fibrosis.

Objective We sought to evaluate the efficacy of bipolar radiofrequency ablation of atrial fibrillation (AF) in patients undergoing cardiac surgery,and to determine independent factors affecting the recurrence.Methods From June 2007 through February 2010,81 patients with atrial fibrillation underwent a modified Cox-Maze Ⅳ procedure using a biopolar radiofrequency device( Medtronic in 32 cases,Atricure in 49 cases).There were 45 males and 36 females,aged (48.2 ± 11.0)years,including 26 cases of permanent AF,44 cases of persistent AF and 11 cases of paroxysmal AF.The mean duration time of AF was (7.56 ± 7.47 ) years.The left atrial diameter were 36 ～ 72 mm.In conjunction with cardiac surgery including:mitral valve replacement (MVR) (or add tricuspid valve plasty (TVP) in 33 cases,mitral and aortic valve replacement (DVR)( or add TVP) in 18 cases,off-pump coronary artery bypass surgery (OPCAB) in 16 cases,aortic valve replacement (AVR) in 4 cases,MVR and coronary artery bypass grafting (CABG) in 1 case,mitral valve plasty (MVP) in 1 case,and others in 8 cases.Among them,22 patients were undergoing electrophysiological mapping by high-frequency bipolar stimulation from June 2009 to February 2010.A follow-up of 12 to 44 months was completed.Recurrences were evaluated by 12-lead ECG or 24 hour Holter recording every clinic visit-1,3,6,9,and 12 months after the procedure and yearly thereafter,or if symptoms developed.Multivariate regression analysis was performed to determine independent factors affecting the recurrence.Results Hospital mortality was 1.23％.The successful ablation of AF were 100％,82.5％,and 84.8％ immediately after operation,at discharge,and at 2(6.1 ± 13.6) months after operation respectively.Multinomial regression analysis showed small left atrium ( ＜ 60 mm),and electrophysiological mapping might contribute better sinus rhythm restoration ( P ＜ 0.05 ).Conclusion Bipolar radiofrequency ablation of atrial fibrillation in patients undergoing cardiac surgery is safe and effective.Small left atrium ( ＜ 60mm) and electrophysiological mapping should be considered to improve results in selected patients.

Objective To observe the clinical efficacy ofQishen Erlian Decoction combined with entecavir for patients with liver fibrosis of hepatitis B.MethodsTotally 74 patients were divided into treatment group (38 cases) and control group (36 cases). The control group was given entecavir, while treatment group was given Qishen Erlian Decoction combined with entecavir, 48 weeks for 1 course. Entecavir was given to both groups after treatment, and two groups were followed up for 24 weeks. The liver function, DNA-HBV, serum TGF-β1, BMP-7, liver fibrosis indexes and clinical symptom changes of the two groups were observed.Results 3 cases were excluded and 3 cases were lost during follow-up in treatment group; 6 cases were lost during follow-up in the control group. Liver function and HBV-DNA in 12, 24, 36, 48 weeks and 24 weeks in follow-up were significantly lower than those pre-treatment (P<0.01) in the treatment group, and were significantly better than those in the control group (P<0.01). TGF-β1 decreased (P<0.05), BMP-7 increased (P<0.01), and the ratio of TGF-β1/BMP-7 decreased (P<0.01) in both groups after treatment. There was significant difference between the two groups (P<0.05). HA, LN, PCⅢ,Ⅳ-C, and FS decreased significantly in treatment group after treatment and in the follow-up (P<0.01), fatigue, discomfort in liver region, disgust oil and anorexia were improved (P<0.05), the difference was significant compared with control group (P<0.05).Conclusion Qishen Erlian Decoction combined with entecavir can not only protect liver and reduce aminotransferase, but also be antiviral and reverse liver fibrosis.

Objective To analyze the status of Leishmania infantum asymptomatic infection in human population of a Kala-azar endemic area in Wenxian County,Gansu Province,and to evaluate the tests used.Methods Blood samples were tested by PCR using two pairs of primers,RV1-RV2 and K13A-K13B,for detecting Leishmania-specific DNA.ELISA and rK39-dipstick were used to detect Leishmania-specific antibodies.Results The positive rate of PCR,ELISA and rK39-dipstick was 30.9%(83/269),24.2%(65/269) and 0(0/269) respectively.Conclusion The prevalence of asymptomatic infection of L.infantum in humans is high in the area.PCR test based on RV1-RV2 and K13A-K13B primer pairs is a sensitive and specific method for detecting the asymptomatic infection.

Objective To discuss the effect of in vitro fertilization ( IVF) and mouse sperm cryopreservation , to establish a simple and economic frozen system for the genetically engineering mice preservation .Methods Sperm from genetically engineering mice were cryopreserved , IVF was performed using post-thawed sperm, then embryo transfer, to compare the effects of cryopreservation medium、age of male mice and sperm preincubation medium .Results Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , the fertility rates were from 82.49%to 91.43%, when HTF was used thawed-sperm preincubation in IVF , the fertility rates were from14.46%to 27.38%, there was a signification difference between PM and HTF sperm preincubation medium;10 to 35 weeks male genetically engineering mice sperm were succeed cryopreservation , and positive mice were procreated after 2-cell embryos were transferred;R18S3、CPM and CPA was used to freeze sperm , the fertility rates were 75.85%、88.89%to 94.27%, positive mice were procreated after 2-cell embryos were transferred;2-cell embryos after IVF were freezed , then thawed and positive mice were procreated after 2-cell embryos were transferred .Conclusion Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , genetically engineering mice sperm were succeed cryopreservation .

The alpha-hemolysin protein of Staphylococcus aureus, which was expressed in Escherichia coli BL21 (DE3) with recombinant pET32a(+)-alpha-HL plasmid, was purified with gel filtration chromatography (GFC) and Ni-NTA spin columns. The quality and biological characteristic were compared. First, the purified products were analyzed with SDS-PAGE, and the expected protein band was with a molecular mass of 53 kD. Second, protein concentration was determined by the method of Bradford, and the median hemolytic dose potency (HD50) was finally analyzed with rabbit erythrocyte. The protein purified with GFC was 0.337 mg/mL, its hemolysis activity was 1519 HU/mg, and hemolysin yield was 14.04%. Meanwhile, the protein purified with the Ni-NTA Spin Columns was 0.35 mg/mL, its hemolysis activity was 1463 HU/mg, and hemolysin yield was 17.5%, respectively. The results showed that there is no significant difference in the quality, hemolysis activity and yield of the recombinant proteins purified with Ni-NTA spin columns and GFC.
Chromatography, Gel ; methods ; Escherichia coli ; genetics ; metabolism ; Hemolysin Proteins ; genetics ; isolation & purification ; Nitrilotriacetic Acid ; analogs & derivatives ; Organometallic Compounds ; Recombinant Proteins ; genetics ; isolation & purification ; Staphylococcus aureus ; genetics ; metabolism

Objective: Colorectal cancer (CRC) is one of the most common cancers and the major cause of cancer death in China. The aim of this study was to estimate the burden of CRC in China. Materials and methods: Data from the National Cancer Center (NCC) of China was used and stratified by area (urban/rural), sex (male/female) for analyzing the age-specific incidence and mortality rates. Time trend of colorectal cancer was calculated based on the 22 high-quality cancer registries in China. National new cases and deaths of colorectal cancer were estimated using age-specific rates multiplied by the corresponding national population in 2014. The Chinese population in 2000 and Segi's world population were used to calculate age-standardized rates of colorectal cancer in China. Results: Overall, 370,400 new colorectal cancer cases and 179,600 deaths were estimated in China in 2014, with about 214,100 new cases in men and 156,300 in women. Meanwhile, 104,000 deaths cases of colorectal cancer were men and 75,600 deaths were women, which accounted for 9.74％ and 7.82％ of all cancer incidence and deaths in China, separately. Relatively higher incidence and mortality was observed in urban areas of China. And the Eastern areas of China showed the highest incidence and mortality. The age-standardized incidence and mortality rate of colorectal cancer has increased by about 1.9％ per year for incidence and about 0.9％ per year for mortality rate from 2000 to 2014. Conclusion: With gradually higher incidence and mortality rate in the past 15 years, colorectal cancer became a major challenge to China's public health. Effective control strategies are needed in China.