RESUMO
Objective:To explore the regulatory effect of miR-873-5p micro-RNA targeting voltage-dependent anion channel protein 1 (VDAC1) in neurons and its mechanism.Methods:Murine nerve cells were randomly divided in vitro into a control group, a model group, a mimetic negative carrier (miR-con) group and an miR-873-5p group. The epileptiform hippocampal nerve cell model was induced in all of the cells except those in the control group using magnesium-free medium. The control group was normally cultured, while the miR-con and miR-873-5p groups were transfected with miR control and miR-873-5p RNA respectively. Real-time fluorescent quantitative polymerase chain reactions were used to detect the expression of miR-873-5p and VDAC1 mRNA. Western blotting was employed to detect VDAC1, B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax) and cleared caspase-3 in the neurons. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) were measured using the DCFH-DA fluorescent probe, the thiobarbituric acid method and enzyme-linked immunosorbent assay respectively. Any apoptosis was detected using flow cytometry, while the targeting of miR-873-5p on VDAC1 was verified using the double fluorescence zymase reporter gene method.Results:Compared with the control group, a significant decrease in the average expression of miR-873-5p, Bcl-2 and in GSH and MDA levels was observed in the model group, but there was a significant increase in the average level of VDAC1, Bax, cleaved caspase-3 and ROS and in the rate of apoptosis. Compared with the miR-con group, a significant decrease in the average expression of Bax, cleaved caspase-3, ROS and in the apoptosis rate was observed in the miR-873-5p group, but there was a significant increase in the average level of Bcl-2, GSH and MDA. Moreover, it was verified that miR-873-5p reduced the expression of VDAC1.Conclusion:miR-873-5p protects damaged neurons by inhibiting their apoptosis through negatively regulating the target gene VDAC1 and the oxidative stress response.
RESUMO
Objective:To observe the clinical efficacy and side effects of injecting different doses of botulinum toxin type A (BTX-A) into children with spastic cerebral palsy (CP) and tiptoe deformity.Methods:A total of 107 children with tiptoe deformity resulting from CP were divided into group A ( n=35), group B ( n=36) and group C ( n=36) using a random number table. Group A received 3u/kg injections of BTX-A, group B received 4u/kg injections and group C received 5u/kg. The injections were guided by color Doppler ultrasound and followed by 4 courses of rehabilitation therapy. Before and 1, 3 and 6 months after the treatment, the modified Tardieu scale (MTS) was used to assess gastrocnemius spasms, while sections D and E of gross motor function scale 88 (GMFM-88) and the pediatric balance scale (PBS) were used to evaluate motor functioning and balance. Any side effects were also observed. Results:After the treatment, improvement was observed in all of the measurements, though there were no significant differences in the degree of improvement nor in the incidence of side effects among the three groups.Conclusions:There is no significant difference in clinical efficacy or side effects involved in using different doses of BTX-A to treat tiptoe deformity in children with spastic cerebral palsy. The recommended dosage is therefore 3u/kg.
RESUMO
The protective effect and mechanism of Schistosoma japonicum cathepsin B (Sjcb2) DNA vaccine in the mouse model of schistosomiasis were studied through construction pcDNA3 .1 (+ ) / Sjcb2 DNA recombinant vector ,which provided effective candidate antigen for anti-schistosome vaccine .The 6-week-old female BALB/c mice were randomly divided into pcDNA3 .1(+ )/Sjcb2 DNA vaccine group ,pcDNA3 .1(+ ) plasmid group and normal saline group ,respectively .Each group was composed of 35 mice ,and 100 μg of S jcb2 plasmid DNA was injected in the hind leg quadriceps of mice once every two weeks .PCR and immunohistochemistry assay were used to detect the expression and stability of Sjcb2 gene in mice .MTT assay was used for testing the specific proliferation response of mice spleen lymphocytes .The level of Sjcb2 antibodies in mouse serum and the IFN-γand IL-4 levels in mice spleen lymphocyte culture supernatant before and after schistosome infection were assayed by ELISA .At last ,we counted load of Schistosome adult worms in mouse and eggs in liver of mouse .The results showed that the Sjcb2 gene was detected in all mice of the Sjcb2 DNA vaccine group ,and Sjcb2 gene expression was positive in the muscle cells in Sjcb2 DNA immunized mice by IHC assay .MTT assay showed that T-cell proliferation rate was in-creased significantly in S jcb2 DNA vaccinated group .ELISA results showed that the IFN-γlevels were increased significantly in the vaccinated group ,while the IL-4 levels were significantly increased after Schistosoma japonicum infection in all mice of every group .The load of worms and eggs in Sjcb2 DNA vaccinated group was reduced significantly than that of control group (P<0 .05) ,the reduction rates of adult worms and eggs were 36 .32% and 60 .61% respectively .In conclusion ,the Sjcb2 gene was stably expressed in muscle cells of mice after injection of S jcb2 recombinant plasmid ,and S jcb2 produced protective effects of anti-schistosoma infection in mice possibly by mean of regulating Th1 cell subgroups through increasing the IFN-γ level and decreasing IL-4 levels .