Porcine reproductive and respiratory syndrome is caused by the PRRS virus(PRRSV), which has six structural proteins(GP2, GP3, GP4, GP5, M and N). GP5 and N protein are important targets for serological detection by enzyme-linked immunosorbent assay(ELISA)and other methods. Toward this goal, we developed an indirect ELISA with recombinant GP5 antigens and this method was validated by comparison to the LSI PRRSV-Ab ELISA kit. The results indicated that the optimal concentration of coated recombinant antigen was 0.2 μg/well for a serum dilution of 1:40. The rate of agreement with the LSI PRRSV-Ab kit was 88.7%(266/300). These results support the potential use of recombinant GP5 as an antigen for indirect ELISA to detect PRRSV antibodies in pigs.

In this study,a panel of monoclonal antibodies (mAbs) against Porcine reproductive and respiratory syndrome virus(PRRSV),named as 8C9 and4B4,were produced by fusing SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with the PRRSV (TCID50=5.5),screened by the indirect ELISA and subjected to several limiting dilutions.mAbs were then identified by biological characterization.Among the two fusion cell strains,8C9 belonged to the IgG1 subclass and 4B4 belonged to the IgG2a subclass.The titers in cell culture supernatant and abdomen liquor reached to 1:104and 1:105,respectively.The specificity test indicated that the two cells had specific reactions for the PRRSV and GP5 protein respectively,and no reaction with Classical swine fever virus (CSFV) or Swine vesicular disease virus (SVDV).The molecular weights of the heavy chain and light chain were about 45.0 kDa and 25.0 kDa,respectively.In neutralization activity tests,the results showed that the prepared mAb 4B4 can protect 50% of cells with no CPE in dilution up to 1:512,but mAB 8C9 has no neutralization activities to PRRSV.

Objective To evaluate the efficacy of regional perfusion of chemotherapeutic drug in rectal cancer patients who had undergone total mesorectum excision(TME).Methods The therapeutic effects were retrospectively analyzed of 178 rectal carcinoma patients,who were admitted in the hospital from 1999 to 2006,had undergone TME,and then received chemotherapy through various routes.Among them,a catheter was placed in the left internal iliac vein,and chemotherapeutic drug was administered through it postoperatively in 90 patients.The other 88 cases served as control,and they received systemic intra-venous chemotherapy after TME.No significant difference on clinical parameters was evident between the two groups during the course of treatment.The 1,3 and 5 year local recurrence rates,metastasis rates and survival rates of the two groups were compared.Results The 1,3 and 5 year local recurrence rates in the study group were 0,2.5%(2/79) and 3.9%(3/77),respectively,while they were 1.1%(1/88),3.9%(3/77) and 8.1%(6/74),respectively in control group.There was a significant difference in the local recurrence rate between two groups(P

Eight mouse hybridoma cell lines which stably secreted monoclonal antibodies ( McAbs ) against human prostate-specific antigen-α1-antichymotrypsin complex ( PSA-ACT ) were obtained through hybridoma technique. After purification, the immunological characters of 8 McAbs were identified and classified by epitopes analysis through indirect enzyme-linked immunosorbent assay ( ELISA) . A pair of McAbs was chosen from above 8 McAbs, based on which a highly sensitive, simple and rapid chemiluminescence enzyme immunoassay ( CLEIA) was developed for determination of PSA-ACT in human serums using the lumino-H2 O2 reaction catalyzed by horseradish peroxidase ( HRP) as the chemiluminescence system. Several experiment factors such as coating buffer, coating concentration, dilution ratio of PSA-ACT-HRP complex, incubation time, immunoreaction protocol and chemiluminescence reaction time were optimized. The results showed that the linear range of the proposed method for PSA-ACT determination was 0-40 ng/mL (R2=0. 9943), with the detection limit of 0. 53 ng/mL. The inter-assay relative standard deviations (RSDs) were 4. 6%-6. 6%, and intra-assay RSDs were 5 . 7%-8 . 0%. The recoveries of PSA-ACT at three spiked levels in serum samples were 95. 4%-104. 2%. The proposed method exhibited a cross-reactivity of 0. 6% with free-PSA. The proposed method is stable, sensitive, rapid and simple, and provides a foundation for the development of PSA-ACT CLEIA kit and shows great value in clinical auxiliary diagnosis of prostate cancer.

Objective To evaluate the role of regional chemotherapy via internal iliac artery pump,systemic chemotherapy and total mesorectum excision(TME) in the treatment of rectal cancer.Methods A total of 193 patients with rectal carcinoma divided into 2 groups: The observation group,included 98 cases who underwent TME,regional chemotherapy through internal iliac artery pump and systemic chemotherapy.The other 95 cases,as control group,were treated with systemic chemotherapy after TME.The local recurrence rate,metastasis rate and survival rate were compared between the two groups.Results The local recurrence rate and metastasis rate of the observation group was significantly lower than that of the control group.The local recurrence rate at 1-,3-and 5-year was significantly lower in the observation group[0,2.5%(2/81) and 3.8%(3/79) respectivvely]than that in the control group [1.1%(1/95),11.4%(9/79) and 16.2%(11/68) respectively](P

Interactions between FMDV and cardiac cells are multifaceted and complex ,these interactions leads to pro-teins alterations in cardiac cells inevitably .To understand the pathogenesis of myocarditis after FMDV infection in mice ,the suckling mouse model for myocarditis induced by foot-and-mouth disease virus (FMDV) was established in this study .Suckling mice within 3 days old was selected to infect by FMDV .Myocarditis caused by FMDV in suckling mice was confirmed with clinical symptom monitor .The observation of Hematoxylin and eosin stain (H&E stain) and transmission electron microscopy (TEM) were performed after samples processing .According to conventional polymerase chain reaction (PCR) methods ,prim-ers of VP1 gene was designed ,synthesised and specific FMDV VP1 gene was amplified from the heart muscle of suckling mice . The results indicated that suckling mice appeared low spirit condition ,dyspnea ,and dull reaction within 36 hours after chal-lenge with FMDV .Infiltration of inflammatory cells and dissolution of myocardial fibers were observed with H&E stain and TEM .Special target gene of FMDV was amplified from the heart of infected group .Obvious inflammation in the heart of suck-ling mice caused by FMDV was observed .It's suggested that suckling mouse model for myocarditis induced by FMDV was es-tablished successfully ,which would lay the foundation for researches of myocarditis mechanism in young cloven-hoofed ani-mals .

Objective To identify the influence of chemotherapy-induced serum alkaline phosphatase (ALP) elevation on the tumor-free survival (TFS) in patients of gastric carcinoma after radical gastrectomy.Methods The clinical data of 189 gastric carcinoma patients receiving radical surgery and postoperative adjuvant chemotherapy between Jan,2010 and Dec,2015 were reviewed and statistically analyzed.Results The TFS of patients with serum ALP elevation was obviously inferior than those without ALP elevation (x2 =5.717,P =0.017),serum ALP elevation was an independent risk factor influencing patients' TFS (HR =2.178,P =0.032),the degree of serum ALP elevation was associated with patients' TFS (x2 =4.627,P =0.031).Conclusion Serum ALP elevation during postoperative chemotherapy indicates the increases of recurrence or metastasis rate of gastric cancer patients after radical gastrectomy.

The beneficial effects of hypoxic preconditioning are abolished in the diabetes. The present study was designed to investigate the protective effects and mechanisms of repeated episodes of whole body hypoxic preconditioning (WBHP) in db/db mice. The protective effects of preconditioning were explored on diabetesinduced vascular dysfunction, cognitive impairment and ischemia-reperfusion (IR)-induced increase in myocardial injury. Sixteen-week old db/db (diabetic) and C57BL/6 (non-diabetic) mice were employed. There was a significant impairment in cognitive function (Morris Water Maze test), endothelial function (acetylcholineinduced relaxation in aortic rings) and a significant increase in IR-induced heart injury (Langendorff apparatus) in db/db mice. WBHP stimulus was given by exposing mice to four alternate cycles of low (8%) and normal air O2 for 10 min each. A single episode of WBHP failed to produce protection; however, two and three episodes of WBHP significantly produced beneficial effects on the heart, brain and blood vessels. There was a significant increase in the levels of brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) in response to 3 episodes of WBHP. Moreover, pretreatment with the BDNF receptor, TrkB antagonist (ANA-12) and NO synthase inhibitor (LNAME) attenuated the protective effects imparted by three episodes of WBHP. These pharmacological agents abolished WBHP-induced restoration of p-GSK-3β/GSK-3β ratio and Nrf2 levels in IR-subjected hearts. It is concluded that repeated episodes of WHBP attenuate cognitive impairment, vascular dysfunction and enhancement in IRinduced myocardial injury in diabetic mice be due to increase in NO and BDNF levels that may eventually activate GSK-3β and Nrf2 signaling pathway to confer protection.

The beneficial effects of hypoxic preconditioning are abolished in the diabetes. The present study was designed to investigate the protective effects and mechanisms of repeated episodes of whole body hypoxic preconditioning (WBHP) in db/db mice. The protective effects of preconditioning were explored on diabetesinduced vascular dysfunction, cognitive impairment and ischemia-reperfusion (IR)-induced increase in myocardial injury. Sixteen-week old db/db (diabetic) and C57BL/6 (non-diabetic) mice were employed. There was a significant impairment in cognitive function (Morris Water Maze test), endothelial function (acetylcholineinduced relaxation in aortic rings) and a significant increase in IR-induced heart injury (Langendorff apparatus) in db/db mice. WBHP stimulus was given by exposing mice to four alternate cycles of low (8%) and normal air O2 for 10 min each. A single episode of WBHP failed to produce protection; however, two and three episodes of WBHP significantly produced beneficial effects on the heart, brain and blood vessels. There was a significant increase in the levels of brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) in response to 3 episodes of WBHP. Moreover, pretreatment with the BDNF receptor, TrkB antagonist (ANA-12) and NO synthase inhibitor (LNAME) attenuated the protective effects imparted by three episodes of WBHP. These pharmacological agents abolished WBHP-induced restoration of p-GSK-3β/GSK-3β ratio and Nrf2 levels in IR-subjected hearts. It is concluded that repeated episodes of WHBP attenuate cognitive impairment, vascular dysfunction and enhancement in IRinduced myocardial injury in diabetic mice be due to increase in NO and BDNF levels that may eventually activate GSK-3β and Nrf2 signaling pathway to confer protection.

We produced beta1 gene which is about 2400 bp by reverse transcription polymerase chain reaction (RT-PCR) from bovine trachea, reclaimed and purified, then cloned the amplified fragment to pGEM-T easy vector, confirmed by sequencing. The immune-dominant epitope of beta1 gene was chosen by computer analysis and then syncretized ligand-binding domain from 346 bp to 843 bp of ecytoplasm with six histidine, expressed LBD protein massly in E. coli BL21 (DE3), and identified by SDS-PAGE. The fusion protein was purified with Ni-NTA affinity chromatography and immunized New Zealand rabbits preparing of its polyclonal antibody, the specific antibody titer was above 1:12,800 detected by indirect ELISA, the result of Western blot showed that this antibody could be recognized by LBD fusion protein.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cattle ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease Virus ; physiology ; Integrin alpha1beta1 ; biosynthesis ; genetics ; immunology ; Ligands ; Protein Binding ; Protein Interaction Domains and Motifs ; Rabbits ; Receptors, Virus ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology