To develop a safe and effective new generation vaccine for IRKV-THChina12 prevention, we constructed a non-replicative recombinant human adenovirus carrying the IRKV-THChina12 G gene, named as rAd5-IRKV-G. The IRKV-THChina12 G protein expressed by the recombinant human adenovirus in 293AD cells was detected by western blot and indirect immunofluorescence test. To evaluate the immunogenicity of the recombinant, mice were immunized with rAd5-IRKV-G by intramuscular (i. m.) or intraperitoneal (i. p.) route and with non-exogenous gene expressing wild type adenovirus wt-rAd5 as a control. Results showed that the rAd5-IRKV-G could induce continuous and statistically significant (P ≤ 0.05) anti-IRKV neutralizing antibody (NA) production in immunized mice by i. m. or i. p. route. In particular, no significant difference (P > 0.05) of the NA titers between the two administration routes were observed, that provides an alternative choice for animal immunization method in the future application.
Adenoviruses, Human ; genetics ; physiology ; Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; GTP-Binding Proteins ; genetics ; immunology ; Gene Expression ; Genetic Vectors ; genetics ; physiology ; Humans ; Immunization ; Lyssavirus ; enzymology ; genetics ; immunology ; Mice ; Rhabdoviridae Infections ; immunology ; virology ; Viral Proteins ; genetics ; immunology ; Virus Replication

Potency is one of the most important indexes of inactivated vaccines.A number of methods have been established to assay the potency,of which the NIH test and single-dose mouse protection test are the prescribed methods.Here,we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine,which uses fewer animals and takes less time to complete.Depending on the quality requirements of a vaccine(e.g.minimum potency),a rabies reference vaccine is,for example,diluted to the minimum potency,and 50 μL of the dilution is taken to inoculate 10 mice.The same amount of the test rabies vaccine is inoculated into another 10 mice.After two weeks,all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization(FAVN) test.By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine,the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality.The reliability of this method was also confirmed in dogs.The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.

0.05).CONCLUSION:The patient with Broca aphasia is independent in the course of the orthographic input,and the ability to retrieve semantic information from orthographic activation word may not affected by phonological lexicon.

Objective To investigate if anodal transcranial direct current stimulation (atDCS) of the right cerebellum improves verbal working memory in amnesic persons with mild cognitive impairment (aMCI).Methods Thirty-nine aMCI were randomly divided into an observation and a control group using a random number table.The observation group was given atDCS at 1.2 mA for 20 minutes every day for 5 days,while the control group was provid ed with fake atDCS in the same way.Before and after the treatment,both groups were tested using forward and back ward digit spans,word reading,visually cued sensorimotor tests and finger tapping.Results After the treatment,the forward and backward digit spans of the observation group had improved significantly compared with before the treatment and with the control group's improvements.Significant improvement was observed in the average backward digit span of the control group,but not in their forward digit span after the treatment.No significant differences be tween the two groups were observed in the other measurements before or after the treatment.Conclusion Direct current stimulation of the cerebellum may improve the verbal working memory deficits of aMCI.Further research should be conducted to find the mechanism.

Objective To test reliability and validity of a verbal behavior assessment scale (VerBAS). Methods The VerBAS was used to evaluate 20 patients with speech disorder repeatedly by the same investigator with a two week interval to assess its reliability. The construct validity of the VerBAS was evaluated by using it to evaluate 235 patients with speech disorder. Results The test-retest correlation coefficient γ was 0.723,which was significant at the 5% confidence level. Cronbach'a a=0.819. Three distinct factors were identified: receptive speech,communicative speech and delineative speech;and their accumulated variance contribution was 83%. Conclusion The Verbal Behavior Assessment Scale had satisfactory reliability and validity, It can be used to evaluate the patients with speech disorder and could provide a reference for speech rehabilitation training.

To test the hypothesis that the controlled reperfusion of warm blood cardioplegiacontaining mannitol would result in more effectively improved recovery of myocardial function by prevent-ing or reducing a potentially harmful component of reperfusion. Methods: Thirty-two cats were divided in-to four groups. Group Ⅰwas not subjected to ischemia or reperfusion injury. Group Ⅱ was subjected to60 min hypothermic ischemia. Group Ⅲwas subjected to 60 min hypothermic ischemia and 60 min reperfu-sion. Group Ⅳ was controlled reperfusion with warm blood cardioplegia containing mannitol. Results:Myocardial functlon was significantly depressed after 60 min reperfusion- Increased myocardial water con-tent and low ATP c0ntent were observed also. Controlled reperfusion with warm bl0od cardioplegia con-taining mannitol was helpful to improve the recovery of myocardial function and ATP content, and to re-duce the myocardial water content. Conclusion: These results indicate that controlled reperfusion after is-chemia provides benefit in avoiding myocardium from reperfusion injury.

Objective To evaluate the efficacy and safety of domestic olmesartan in treatment of mild to moderate essential hypertension in comparison with losartan. Methods Two hundred and thirty-seven patients with mild to moderate essential hypertension were enrolled in a randomized, double-blind, multi-center, paralleded and active-controlled trial, and were divided into olmesartan group (olmesartan 20 mg + losartan 50 mg placebo) and losartan group (losartan 50 mg + olmesartan 20 mg placebo) for a 8-week therapy. Four weeks after treatment, dosages of drugs were doubled in patients with seated diastolic blood pressure ≥90 mmHg (1 mmHg =0.133 kPa). All patients were followed up every two weeks, and the efficacy and adverse effects were observed. Another 32 patients with mild to moderate essential hypertension were enrolled and given olmesartan only, and ambulatory blood pressure monitoring was performed before and 8 weeks after treatment. Results Compared with those before treatment, both systolic blood pressure and diastolic blood pressure significantly decreased in olmesartan group and losartan group 8 weeks after treatment [(15.2 ±13.3) mmHg and (19.5 ±11.8) mmHg, respectively for systolic blood pressure (P <0.001); (15.9 ±7.48) mmHg and (16.2 ± 5.95) mmHg, respectively for diastolic blood pressure (P < 0.01) ], while there was no significant difference between these two groups (P > 0.05). There was no significant difference in total effective rate and incidence of adverse effect between these two groups (86.9% vs 93.7% and 7.63% vs 5.88% , P > 0.05) . Ambulatory blood pressure monitoring demonstrated that trough to peak ratios of systolic blood pressure and diastolic blood pressure were 86% and 71%, respectively. Conclusion Domestic olmesaratan provides an effective, safe and long action in the treatment of mild to moderate essential hypertension.

Objective To test the immune therapeutic effects of PICKCa adjuvant rabies vaccine for the post-exposure mice and beagle dogs and immune memory effect for human.Methods For the mice and beagle dogs first to infect with rabies wild virus and then immunize with PICKCa rabies vaccine.For volunteers, first to immunize with PICKCa rabies vaccine after 2.5 years to take peripheral blood mononuclear cell and then to check cells secreted interferon-gwith flow cytometry. Results In 3 independent assays of post-explosure immunization of mice and beagle dogs PICKCa rabies vaccine were much better then commercial adjuvant-free rabies vaccines, the protective rate from 20%-30%to 70%-100%Also, in the assay of cells secreted IFN-γby peripheral blood mononuclear cells from volunteers immunized by PICKCa rabies vaccine the IFN-γcell levels were much higher than control.Conclusions Compared with commercial adjuvant-free rabies vaccines the PICKCa rabies vaccine had significant immune therapeutic effects in the mice and dogs.Also PICKCa rabies vaccine had good immune memory effect in human.

African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.

In order to obtain a cell line which high expressed of rabies virus glycoprotein the produc-tion of rabies subunit vaccine,the RABV glycoprotein gene sequence was amplified by RT-PCR and cloned into the lentiviral expression vector.The recombinant plasmid pLV-RVG,envelope plasmid pMD2.0G and packaging plasmid psPAX2 were co-transfected into HEK-293T cells to harvest lentivirus and infect new HEK-293T cells.The 293T-RVG cell line was obtained by puro-mycin pressure screening.Indirect immunofluorescence assay(IFA)and Western blot showed that the 67 kD RABV-G protein was highly expressed in the cells.The neutralizing antibody titer reached 2.19 IU/mL within 14 days after immunization,which was higher than the internationally recognized protection threshold(0.5 IU/mL).This study showed that the cell line 293T-RVG with stable and high expression of rabies virus glycoprotein was successfully constructed.The cell line could protect mice against rabies virus challenge and could be used as a subunit vaccine with poten-tial for large-scale production and application.