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BACKGROUND:Induced pluripotent stem cel technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE:To comprehensively review the safety and application of induced pluripotent stem cel s.METHODS:A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of“induced pluripotent stem cel , reprogramming, clinical application, safety, transcription factor, disease mode”in English and Chinese, respectively. RESULTS AND CONCLUSION:In recent years, research on induced pluripotent stem cel s has attracted much attention from the scientific community and the medical community, and this technique has successful y gained induced pluripotent stem cel s and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cel s. Therefore, we are faced with enormous difficulties and chal enges, which involve al aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cel s into the desired cel type and how to establish a suitable disease model as wel as a high-throughput drug screening platform.
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Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .
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Objective To investigate the effects of exercise on expression of microtube associated protein-2 (MAP-2) and MAP-2 mRNA in neuronal cells after intracerebral hemorrhage (ICH) in rats. Methods Ninety-six male Sprague-Dawley rats (weighing 270 to 300 g) were divided into 3 groups, a trial group (ICH-induction and ex-ercise, n=32), a control group (ICH-induction only, n=32) and a sham-operated group (no ICH and no exercise, n=32). The brains of the rats were sampled at 7, 14, 21, and 28 days after the operation for establishing ICH mod-el. Another 64 rats were divided into 8 groups (6 h, 12 h, 24 h, 48 h, 72 h, 7 d after ICH, no ICH group and nor-mal group). MAP-2 and MAP-2 mRNA activity were measured by immunohistochemical methods and RT-PCR. The rats in the trial group began cage-running exercise 72 h after the operation. The others were reared in the standard ca-ges. Results ①MAP-2-positive cells appeared around the hematoma in the cortex. The number of MAP-2-positive cells was very small in the sham-operation group. There was an up-regulation of MAP-2 from 24 h to 7 d after ICH, and significant difference was found between the non-ICH group and the normal group. The expression of MAP-2 showed an up-regulation trend in the trial and control groups. There was a significant difference compared with the sham operated group, and the trial group had significantly higher expression levels than the control group. ②RT-PCR showed up-ragulation of MAP-2 mRNA 12 to 24 h after ICH. There was a significant difference between the no ICH and normal groups. The trial and control groups showed up-regulation of MAP-2 mRNA at 14 to 28 d after ICH, with was significantly different from that of the sham group, and the trial group had significantly higher levels than the con-trol group. Conclusion MAP-2 might participated in neural cells plasticity after ICH, and exercise training (cage-running) can up-regulate MAP-2 and MAP-2-mRNA expression.