Grass carp plays an important role in small-scale aquaculture in Vietnam. However, a severe disease, known in Vietnam as "Red Spot Disease", is causing significant economic loss in grass carp aquaculture. In...Grass carp plays an important role in small-scale aquaculture in Vietnam. However, a severe disease, known in Vietnam as "Red Spot Disease", is causing significant economic loss in grass carp aquaculture. In this study, the tissue samples isolated from the grass carp with Red Spot Disease in Vietnam are investigated and comparied with the control GCHV isolated in China by experimental infection, culture cell infection, serological cross reactivity, and RT-PCR amplification. Infected grass carp exhibits hemorrhage symptoms about 5 days after experimental injection with GCHV-V (Vietnam) strain. The symptoms and lethality induced by the GCHV-V strain are identical to that induced by the Chinese GCHV-9014 strain. The Chinese GCHV-873 strain induces typical cytopathogenic effects in 4 cell lines, such as CIK, CAB, FHM and GCO, from the 6 fish cell lines examined. No cytopathogenic effects are observed in all the 6 examined cell lines, including CAB, FHM, CIK, EPC, CCO and GCO, infected by the GCHV-V strain and GCHV-9014 strain. Immunodiffusion assays demonstrate an obvious cross-reactivity among three GCHV strains. Precipitin lines are clearly observed not only between the anti-GCHV-873 serum and the two strains GCHV-873 and GCHV-9014, but also between the anti-GCHV-873 serum and the GCHV-V strain. GCHV can be detected by immunodiffusion assays after three generations of blind propagations in the cell lines inoculated by GCHV-V strain. This implicates that GCHV-V viruses have been replicated and amplified despite there being no cytopathogenic effects observed in these examined cell lines. Three genome segments of GCHV, including S8, S9 and S10, are amplified by three sets of PCR primers designed according to the segment sequences published recently. The Q8fp and Q8rp primer set specific for genome segment S8 amplifies a 955 bp fragment from the extracted sample of diseased fish with Red Spot Disease, and the fragment size is identical to that amplified by the same primer set from control GCHV-873 strain. Simultaneously, the Q9fp and Q9rp primer set specific for genome segment S9 generates a same 635 bp product, and the Q10fp and Q10rp primer set specific for genome segment S10 produces a same 697 bp fragment from both template samples of diseased fish with Red Spot Disease and control GCHV-873 strain. The RT-PCR amplification and corresponding size comparison data indicate that the three GCHV-V genome segments extracted from the diseased grass carp with Red Spot Disease in Vietnam should be identical to that in control GCHV-873 strain from China. The data confirm that the causative agent of grass carp Red Spot Disease in Vietnam is a virus, and the virus is closely similar to GCHV strain in China.展开更多
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 ...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.展开更多
Wang Shi Bao Chi Wan(WSBCW)is a traditional Chinese medicine with a recorded administration history of more than 180 years.In the present study,the preclinical safety of WSBCW was evaluated the preclinical safety of W...Wang Shi Bao Chi Wan(WSBCW)is a traditional Chinese medicine with a recorded administration history of more than 180 years.In the present study,the preclinical safety of WSBCW was evaluated the preclinical safety of WSBCW using a toxicity test,which consisted of an administration period of 28 d and a recovery period of 15 d.During the test,male and female SD rats were administered the medicine once a day by oral gavage,at a dose of 60 mg/kg/day,600 mg/kg/day,or 1500 mg/kg/day.As a reference medicine,mosapride citrate was administered at a dose of 37.5 mg/kg/day,which was clinically equivalent to the high-dosage treatment of WSBCW.With all the dosage groups,statistically,no adverse effect was observed in terms of clinical observation,food intake,body weights,organ coefficient,blood biochemistry,and histopathology examination.No intestinal melanosis was observed in the rats.When the data were examined animal by animal,test substance-related adverse effects were found with the high-dosage rats in hematology assay.The deranged,however,reversible changes suggested a compromised intestinal barrier,which was also observed with in mosapride citrate-treated rats.In addition to the histopathology assay,molecular toxicology was explored using high-throughput gene sequencing.No evident toxicity was revealed.In summary,administration of WSBCW was well tolerated within a treatment of 28 d.展开更多
基金This research was supported by National 863 High Technology Research Foundation of China(2002AA62601)National Natural Science Foundation of China(30170726)+1 种基金the Project of Chinese Academy of Sciences(KSCXZ-SW-302)the Innovation Project of the Institute of Hydrobiology,Chinese Academy of Sciences.
文摘Grass carp plays an important role in small-scale aquaculture in Vietnam. However, a severe disease, known in Vietnam as "Red Spot Disease", is causing significant economic loss in grass carp aquaculture. In this study, the tissue samples isolated from the grass carp with Red Spot Disease in Vietnam are investigated and comparied with the control GCHV isolated in China by experimental infection, culture cell infection, serological cross reactivity, and RT-PCR amplification. Infected grass carp exhibits hemorrhage symptoms about 5 days after experimental injection with GCHV-V (Vietnam) strain. The symptoms and lethality induced by the GCHV-V strain are identical to that induced by the Chinese GCHV-9014 strain. The Chinese GCHV-873 strain induces typical cytopathogenic effects in 4 cell lines, such as CIK, CAB, FHM and GCO, from the 6 fish cell lines examined. No cytopathogenic effects are observed in all the 6 examined cell lines, including CAB, FHM, CIK, EPC, CCO and GCO, infected by the GCHV-V strain and GCHV-9014 strain. Immunodiffusion assays demonstrate an obvious cross-reactivity among three GCHV strains. Precipitin lines are clearly observed not only between the anti-GCHV-873 serum and the two strains GCHV-873 and GCHV-9014, but also between the anti-GCHV-873 serum and the GCHV-V strain. GCHV can be detected by immunodiffusion assays after three generations of blind propagations in the cell lines inoculated by GCHV-V strain. This implicates that GCHV-V viruses have been replicated and amplified despite there being no cytopathogenic effects observed in these examined cell lines. Three genome segments of GCHV, including S8, S9 and S10, are amplified by three sets of PCR primers designed according to the segment sequences published recently. The Q8fp and Q8rp primer set specific for genome segment S8 amplifies a 955 bp fragment from the extracted sample of diseased fish with Red Spot Disease, and the fragment size is identical to that amplified by the same primer set from control GCHV-873 strain. Simultaneously, the Q9fp and Q9rp primer set specific for genome segment S9 generates a same 635 bp product, and the Q10fp and Q10rp primer set specific for genome segment S10 produces a same 697 bp fragment from both template samples of diseased fish with Red Spot Disease and control GCHV-873 strain. The RT-PCR amplification and corresponding size comparison data indicate that the three GCHV-V genome segments extracted from the diseased grass carp with Red Spot Disease in Vietnam should be identical to that in control GCHV-873 strain from China. The data confirm that the causative agent of grass carp Red Spot Disease in Vietnam is a virus, and the virus is closely similar to GCHV strain in China.
基金the China National"863"Program(Approval No.2011AA10A212)Special Fund for Agro-Scientific Research in the Public Interest(ApprovalNo.201203056)
文摘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.
基金Beijing Municipal science&Technology Commission(Grant No.2161100001816008)the National Natural Science Foundation of China(Grant No.31571403)Beijing Natural Science Foundation(Grant No.2171001)。
文摘Wang Shi Bao Chi Wan(WSBCW)is a traditional Chinese medicine with a recorded administration history of more than 180 years.In the present study,the preclinical safety of WSBCW was evaluated the preclinical safety of WSBCW using a toxicity test,which consisted of an administration period of 28 d and a recovery period of 15 d.During the test,male and female SD rats were administered the medicine once a day by oral gavage,at a dose of 60 mg/kg/day,600 mg/kg/day,or 1500 mg/kg/day.As a reference medicine,mosapride citrate was administered at a dose of 37.5 mg/kg/day,which was clinically equivalent to the high-dosage treatment of WSBCW.With all the dosage groups,statistically,no adverse effect was observed in terms of clinical observation,food intake,body weights,organ coefficient,blood biochemistry,and histopathology examination.No intestinal melanosis was observed in the rats.When the data were examined animal by animal,test substance-related adverse effects were found with the high-dosage rats in hematology assay.The deranged,however,reversible changes suggested a compromised intestinal barrier,which was also observed with in mosapride citrate-treated rats.In addition to the histopathology assay,molecular toxicology was explored using high-throughput gene sequencing.No evident toxicity was revealed.In summary,administration of WSBCW was well tolerated within a treatment of 28 d.
