Objective:To prepare the monoclonal antibodies against chicken CARP.Methods:The chicken CARP cDNA encoding the N-terminal 110aa length of encoding region was cloned into prokaryotic expression vector pET28b.The BALB/c mice were immunized with purified the recombinant protein,spleen cells of mouse were fused with SP2/0,and the positive clones were screened by indirect ELISA.The properties of McAb were analyzed by ELISA and Western blot.Results:Two hybridomas were proved to have the capability of stably secreting McAb against chicken CARP,designated as 4A8 and 4E6,the subtype of both was IgG1,and the light chain was ?.The titers of ascites were 3.2?104 and 1.28?105 respectively.The Western blot results suggested that they could recognize chicken CARP protein.Conclusion:Two murine hybridomas secreting anti-chicken CARP McAbs have been successfully obtained.

The bacterium Streptomyces peucetius produces doxorubicin and daunorubicin,and the two widely used anticancer antibiotics.Two open reading frames,drrA and drrB,were proposed to encode for an ABC (ATP-binding cassette) type of permease that carries out export of the antibiotic ABC (ATP binding cassette)-type transporter for the exportation of these two antibiotics.In this paper,the structure,expression and interaction of DrrA and DrrB protein and the research progress of the assembly and multi drug efflux function of DrrAB are briefly reviewed.

Background and purpose: ERCC2 gene silence by siRNA interference was observed in esophageal cancer cell line and it could change cell sensitivity to taxol. This study was to investigate the biological mechanism of paclitaxel-resistance in esophageal cancer cells. Methods: ERCC2 targeting siRNA (si-ERCC2) has been synthesized. The constructor were transfected into ERCC2 cell lines high-KYSE150(bigh expression of ERCC2) through lipofectamine. RT-PCR and flow cytometry (FCM) were used to detect the ERCC2 mRNA and protein expression levels. MTr assay was used to estimate the paclitaxel sensitivity of the cells. Results: In si-ERCC2 group, ERCC2 could not be detected and ERCC2 protein expression were reduced by 31.2%, 51.6% and 60.0%, respectively, 24,48,72 b after transfection. Paclitaxel IC_(50) value for si-ERCC2 group was 6.32±0.87 μg/mL, lower than the control group(49%). Conclusion: siRNA could successfully silence the target gene ERCC2 at the level of transcription and translation of the gene, the reduction of ERCC2 expression may reverse taxol resistance of the cells.

Objective:To silence ERCC2 (excision repair cross-complementing rodent repair deficiency,complementatin group 2,ERCC2) expression in esophageal cancer KYSE150 cells by small interfering RNA (siRNA) and observe the altered sensitivity of KYSE150 cells to paclitaxel (PTX) and elucidate the mechanism underlying the reversion of the PTX resistance of KYSE150 cells. Methods:ERCC2-targeted siRNA was synthesized in vitro and transiently transfected into ERCC2 overexpressing KYSE150 cells via Lipofectamine mediation. The mRNA and protein expression levels of ERCC2 were determined by using RT-PCR and FCM method, respectively. The sensitivity of KYSE150 cells to PTX was measured by MTT assay before and after siRNA transfection. Results:RT-PCR results suggested that the specific bands of ERCC2 mRNA were not detected in si-ERCC2 group at 24, 48 and 72 h post transfection. FCM results indicated that the expression levels of ERCC2 protein gradually decreased by 31.2%, 51.6% and 60.0% at 24, 48 and 72 h respectively(P<0.01). The IC_(50) value of PTX for ERCC2-silencing KYSE150 cells was (6.32±0.87) μg/mL, lower than that for control cells (P<0.01). Conclusion:siRNA successfully silenced the expressions of target gene ERCC2 at both the transcription and translation levels. Silencing ERCC2 expression partly reversed the resistance of KYSE150 cells to PTX.

Objective To retrospectively analyze echocardiography findings and left hearst function in patients with patent ductus arteriosus (PDA) after transcatheter closure. Methods 28 patients admitted between January 2012 and December 2012 in our hospital for PDA transcatheter closure were included. Assessment of cardiac structure, hemodynamics and cardiac function parameters during preoperation and in postoperation 3 days, 1 month and 6 months were studied. Results Statistical significant difference was found at 3 days, 1 month and 6 months postoperation when compared with pre-operation in terms of left atrial diameter, left ventricular end-diastolic diameter, left ventricular end systolic diameter and other doppler measurements ( all P < 0. 05 ) . Six-minute walk test ( 6MWT) tolerance improved when compared to preoperation level (P < 0. 05). Better improvement in LVEF and LVES was observed in patients age ≤14 years old when compared to patients > 14 years old after operation ( P < 0. 05 ) . Conclusions Transcatheter closure of PDA can improve cardiac function and correct early hemodynamic abnormalities patients in younger age group show more benefit from the procedure.

In order to characterize the Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity, immunogenicity and immunoprotection of the Staphylococcus aureus (S. aureus) surface protein GapC, gapC gene of S. aureus was amplified from strain BMSA/855/23-1 by PCR, and was inserted into pQE-30 vector subsequently. The recombinant plasmid, designated as pQE/gapC, was transformed into E. coli strain M15 (pREP4). The recombinant GapC fusion proein was successfully expressed in E. coli M15 induced with IPTG and its GAPDH activity was confirmed by GAPDH activity assay. Then, the recombinant GapC protein, inactivated S. aureus whole cell and placebo (PBS) were administrated to healthy rabbits respectively. The IgG antibody titers, concentration of IFN-gamma and IL-4 cytokines in immunized rabbit sera were measured with Enzyme-Linked Immunosorbnent Assay (ELISA). Finally, immunized rabbits were challenged with S. aureus strain Wood46 to evaluate the immunoprotection. The IgG antibody titers against GapC and whole cell in rabbit sera reached their peaks at day 28 after boost immunization (1:64,000). The concentration of IL-4 and IFN-gamma in GapC groups rabbit sera increased significantly (P<0.05) at day 14 after boost immunization, while the concentration of those in whole cell group did not increase (P>0.05) compared with the placebo group. 4 rabbits in 5 of the protein immunized group were protected against challenge with 1 x 10(8) CFU S. aureus. The results above indicate that the expressed recombinant GapC protein have high GAPDH activity and immunogenicity, can also protect against S. aureus challenge to some extent. S. aureus GapC protein could be an attractive target for further genetic engineering vaccine.
Animals ; Antibodies, Bacterial ; blood ; Antigens, Bacterial ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glyceraldehyde-3-Phosphate Dehydrogenases ; biosynthesis ; genetics ; immunology ; Immunization ; Male ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Staphylococcal Vaccines ; immunology ; Staphylococcus aureus ; enzymology ; genetics ; immunology ; Vaccines, Synthetic ; immunology

In order to characterize the immunogenicity and immunoprotection of the Staphylococcus aureus (S. aureus) surface Isdb, we amplified Isdb gene from S. aureus Wood46 strain. The isdb gene was subsequently inserted into pET32a(+) vector and the recombinant plasmid was transformed into E. coli strain BL21. The recombinant Isdb was expressed and purified. Then, we immunized mice with the purified recombinant protein. The antibody level was measured by enzyme-linked immunosorbent assay. Finally, immunized mice were challenged with S. aureus strains Wood46 and HLJ23-1. These results showed that isdb gene sequences were highly conserved, and the recombinant Isdb was successfully expressed. The antibody titer in the immunized groups was increased significantly (P < 0.05) compared with the control, the protective rate of Isdb protein inducted by challenge with the two S. aureus stains Wood46 and HLJ23-1 was 62.5% and 75%, respectively. These results showed that the Isdb protein had high immunogenicity and immunoprotective capacity.
Animals ; Antibodies, Bacterial ; blood ; Cation Transport Proteins ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; genetics ; Immunization ; Male ; Mice ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Staphylococcal Infections ; immunology ; prevention & control

ObjectiveTo assess the effectiveness of tele-medicine and self-management goal(SMG) setting technique used in the diabetes management in the community setting.Methods It is a control-group study.415 type 2 diabetic residents were recruited from the Shanghai Wuliqiao community based on existing medical records.The subjects were divided into two groups,the study group was cared by general practitioners (GPs) specialists cooperation through the tele-medicine mechanism,the other was a control group.For the study group,a cooperation pathway between community health care centers and general hospitals were established.Standardized training and guidelines were provided to community health workers,regarding the setting of management goals of blood glucose and blood pressure,treatment plan,patient education,and SMG techniques.Fasting blood glucose ( FBG ) and 2 h postprandial blood glucose (2hBG) in the study group were monitored,followed by community health workers visiting monthly with seminars for diabetes education.At the baseline and the 12tb month,FBG,2hBG,HbA1C,blood pressure,triglyceride,total cholesterol,body mass index,waist-hip ratio were determined in each group.A survey was conducted to evaluate the costs of diabetes treatments,the knowledge base related to their disease,lifestyle,and the awareness of the new care model.The rates of achieving the goal of blood glucose,blood pressure,and HbA1Ccontrol were calculated.Internet case discussion between GPs-Specialists and referral to certain specialists were implemented when some patients did not reach the control goal.ResultsBy the 12 month follow up,FBG,2hBG,HbAIc,blood pressure of the study group were lower than the baseline,and as well as the control group with statical significance (P＜0.05).There are other improvcments:diabetes knowledge (29.1％ vs 5.5％ ),healthy diet (9.6％ vs -10.4％ ),blood glucose monitoring (30.3％ vs 10.8％ ),support for diabetes care in community (35.7％ vs 9.4％ ),and the preference of the new model (63.8％ vs 17.9％ ) with statistical significauce (P＜0.01 ).As for the medical costs,the study group's monthly costs were consistently lower than the control's.( -3.39Yuan vs 32.26 Yuan,P＜0.05).ConclusionsThe new diabetes care model based on GPs-Specialists tele-medicine and SMG in community opens the door to the community based care model formulation in regard to the health quality and costs control.The deployment of more technologies and management techniques could be explored further to improve the outcomes of community based chronic disease care model.

In order to characterize the immunogenicity and immunoprotection of the Staphylococcus aureus (S. aureus) surface protein Clumping factor A (ClfA), we amplified clfa genes from S. aureus Newman strain, Wood46 strain and HLJ23-1. The clfa gene from Newman strain was subsequently inserted into pQE-30 vector and the recombinant plasmid was transformed into Escherichia coli strain M15 (pREP4). The recombinant ClfA protein was expressed and purified. Then, we immunized mice with the purified recombinant protein. The antibody level and the concentration of cytokines were measured by enzyme-linked immunosorbent assay. Finally, immunized mice were challenged with S. aureus Newman, Wood46 and HLJ23-1. These results suggested that clfa gene sequences were highly conserved, and the recombinant ClfA was expressed correctly with good antigenicity. The antibody titer and the concentration of cytokines in the immunized groups increased significantly (P < 0.05) compared with control, and the mice in the immunized groups were protected against the challenge strains to some extent. These results showed that the ClfA had high immunogenicity and immunoprotective potential.
Animals ; Coagulase ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Immunization ; Mice ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Staphylococcus aureus ; metabolism ; pathogenicity

The clinical data of a 64-year-old patient with adrenocortical adenoma complicated with inferior vena cava tumor thrombus(IVCTT) were retrospectively analyzed. The patient was admitted becourse of intermittent dizziness for 4 months. CT examination revealed right adrenal tumor, and IVCTT was found in operation. Adrenal cortical adenoma needs to be distinguished from adrenal cortical carcinoma pathologically. Preoperative color Doppler ultrasonography, CT angiography or inferior vena cava angiography can confirm the diagnosis of IVCTT and tumor thrombus grade, and different surgical methods should be selected according to tumor thrombus grade.