Objective To establish a novel method by integrating immunomagnetic bead enrich-ment with immunochromatography for the detection of influenza A virus. Methods The immunomagnetic beads were prepared by using EDC/NHS method and then coupled with monoclonal antibodies against influ-enza A virus. A direct immunomagnetic beads-based immunochromatography for the detection of influenza A virus was developed by using double-antibody sandwich method and immunochromatography, which was fur-ther combined with immunomagnetic separation to establish the novel integrated method of immunomagnectic bead enrichment and immunochromatography. Clinical throat swab samples collected from patients with influ-enza A virus infection and healthy subjects were analyzed by the novel method and the results were compared with those by using the conventional colloidal gold immunochromatography to evaluate the specificity, sensi-tivity and positive coincidence rate of this established method. Results The direct immunomagnetic beads-based immunochromatography and the colloidal gold immunochromatography showed no significant differences in specificity and sensitivity and could be used to identify influenza A virus-positive samples with cycle threshold ( Ct) values less than or equal to 22 obtained by real-time PCR assay. The integrated method could identify positive samples with Ct values less than or equal to 28, indicating that the novel method was more sensitive. Conclusion The novel method by integrating immunomagnetic bead enrichment with immunochroma-tography was successfully established and suitable for the rapid and on-site detection of influenza A virus.

BACKGROUND:Breviscapine treatment of cerebral infarction has curative effects, few side effects, stable long-term efficacy and few side effects, which can improve the micro-environment of damaged central nervous system after cerebral infarction.
OBJECTIVE:To investigate the effects of breviscapine injection combined with bone marrow mesenchymal stem cel transplantation on neurological recovery and growth-associated protein 43 expression in rats after cerebral infarction.
METHODS:Sixty Sprague-Dawley rats undergoing middle cerebral artery occlusion were randomized into cerebral infarction group, cel transplantation group and combined group. At 6 hours after modeling, 1 mL PBS, 1 mL bone marrow mesenchymal stem cel suspension (2.5×106), and 1 mL bone marrow mesenchymal stem cel suspension (2.5×106)+75 mg/kg breviscapine injection were respectively injected via the tail vein in the three groups, once a day, continuously for 5 days.
RESULTS AND CONCLUSION:At 2 weeks after transplantation, BrdU-positive bone marrow mesenchymal stem cels were mainly gathered in the peri-infarction region, and the number of positive cels was higher in the combined group than the other two groups (P < 0.01). At 1, 2, 3 weeks after transplantation, the neurological deficit scores were significantly lower in the combined group than the other two groups (P < 0.05). At 2 weeks after transplantation, the combined group had smaler infarct size, milder edema, and higher expression of growth-associated protein 43 as compared with the other two groups (P < 0.05). Under light microscope, glial cels proliferated dramaticaly and brain edema significantly reduced in the combined group. These findings indicate that breviscapine injection combined with bone marrow mesenchymal stem cel transplantation can significantly reduce infarct size and brain edema, promote neurological recovery and increase the expression of growth-associated protein 43 in rats with cerebral infarction.

Objective To investigate the effect and mechanism of FTY720 in protecting the motor nerve function in rats with acute spinal cord injury (ASCI).Methods A model of ASCI in the rat was established using Allen' s method.The rats were divided into five groups according to random number table:Group A (normal control group,n =6),Group B (sham group,n =27),Group C (ASCI group,n =27),Group D (FTY720 systemic treatment group,n =27) and Group E (FTY720 topical treatment group,n =27).After treatment,each group was observed for the changes of lymphocytes in peripheral blood and topical spinal cord tissue,BBB score,pathological changes of spinal cord tissue and expression intensity of glial fibrillary acidic protein (GFAP).Results After ASCI in rats,the infiltration of local T lymphocyte subpopulations CD3,CD4 and CD8 was promoted,but the number of peripheral blood T lymphocyte subpopulations CD3,CD4 and CD8 was decreased.After FTY720 therapy,both local and peripheral blood lymphocytes were reduced in number and the decrease of peripheral blood lymphocytes in Group E was between Groups C and D.BBB score,pathologic observation and GFAP staining showed a neuroprotective effect of FTY720 and better motor nerve function recovery in Group E than in Groups C and D.Conclusions FTY720 therapy facilitates motor nerve function recovery.Local FTY720 application reduces peripheral blood lymphocytes and lowers the incidence of infection.

A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV.PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.

Objective: To observe the clinical therapeutic effect of Tang Shen Ning(TSN,a prescription of traditional Chinese medicine)for curing the early stage of diabetic nephropathy(DN).Methods: Adopting random and double-blind controlled trial methods,several indexes were observed in terms of uric micro-amount albumin excretion rate,creatinine clearance,hemorheology and platelet function et al.Results: TSN could significantly reduce urinary albumin,glomerulus's hyper-filtration,improve blood stream transform and inhibit platelet aggregation.Conclusion: TSN could protect renal functions during the early stage of DN.