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AIM:To investigate the therapeutic effect of mitochondrial fission inhibitor-1(Mdivi-1)on experi-mental autoimmune encephalomyelitis(EAE)in mice,and to explore its mechanism.METHODS:The mice immunized with myelin oligodendrocyte glycoprotein peptide fragment 35-55(MOG35-55)were randomly divided into DMSO model group and Mdivi-1 intervention group.All mice were sacrificed on the 28th day after the first immunization.The demyelination was analyzed by Luxol fast blue staining.The protective mechanism of Mdivi-1 in the spinal cord tissue was investigated by immunofluorescence staining,TUNEL staining and the in vitro experiment with MO3.13 oligodendrocytes treated with staurosporine.The mitochondrial depolarization was detected by JC-1 staining,the cell injury was checked by LDH leakage,and the viability of MO3.13 oligodendrocytes was determined by MTT assay.RESULTS:Compared with DMSO model group,the demyelinating injury was alleviated and the proportion of apoptotic CC1+ oligodendrocytes in Mdivi-1 group was decreased.The cleaved caspase-3,caspase-9,cytochrome C and Bax protein expression levels in the spinal cord of Mdivi-1-treated mice was also attenuated.The in vitro MO3.13 cell experiments suggested that Mdivi-1 inhibited MO3.13 cell mitochondrial depolarization,attenuated the cell damage and increased the cell viability.CONCLUSION:Mdivi-1 pro-tects against the myelin injury in EAE mice,which may be related to the suppression of oligodendrocyte apoptosis.
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【Objective】 To explore the effect and mechanism of Fasudil in the treatment of experimental autoimmune myocarditis (EAM) in mice so as to provide a theoretical basis for the clinical use of Fasudil in treating myocarditis. 【Methods】 Balb/c male mice were used as the research objects, and the EAM mice model was constructed using MyHC-α614-629 polypeptide. Mononuclear cells were isolated and cultured to detect the number of mononuclear cells in mouse spleen. Inflammation infiltration, fibrosis and IL-6 expression in mouse myocardial tissue were detected by HE staining, Masson staining and immunohistochemistry, respectively. The protein expressions of Notch1 and IL-6 were detected by Western blotting. qRT-PCR was used to detect the expressions of pro-inflammatory factors (IL-1α, IL-1β and IL-6) as well as key genes of TLRs and NOTCH signaling pathway. 【Results】 EAM mice showed increased HW, decreased BW, increased HW/e-BW, and increased inflammatory infiltration and fibrosis in myocardial tissue. The above-mentioned symptoms or pathological features were improved in EAM mice treated with Fasudil. The analysis showed that the pro-inflammatory factors IL-1α, IL-1β and IL-6 in the myocardial tissue of EAM mice were significantly increased, but only the expression of IL-6 was statistically different after Fasudil treatment compared with the control group. In addition, TLRs signaling pathway might also play an important role in the EAM mice treated with Fasudil. The expressions of IL-6 and Notch1 were consistent, and the expressions of the key genes of NOTCH signaling pathway (Notch1, Hes1 and Jag2) were down-regulated after Fasudil treatment. 【Conclusion】 Fasudil exerts a protective effect on down-regulation of IL-6 expression by inhibiting the NOTCH signaling pathway in EAM mice.
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Objective:To investigate the effects of silencing HOXA13 gene on the malignant phenotypes of hepatocellular carcinoma cell lines HepG2and QGY-7703,and to provide a new molecular target for the diagnosis and treatment of hepatocellular carcinoma.Methods:The interference recombinant plasmid of plent-U6-GFP/si-HOXA13 was constructed,then it was stably transfected into the HepG2and QGY-7703 cells.The interfering effects of HOXA13 gene were determined by RT-PCR and Western blotting methods.The HepG2and QGY-7703 cells transfected with HOXA13 knockdown were used as experimental group,and the HepG2and QGY-7703 cells transfected with empty plasmid were used as control group.Then the growth speed was examined by MTT assay, the cell doubling time was examined by double time assay,the colony formation ability was examined by colony formation assay,and the cell cycle was examined by flow cytometry.Results:The MTT assay results showed that compared with control group,the growth speeds of HepG2and GY-7703 cells in experimental group were significantly decreased and the cell cycles were changed;the number of HepG2and QGY-7703 cells in G1phase was increased and the number of cells in S phase was decreased.The doubling time of HepG2and QGY-7703 cells in control group and experimental group were(4.59±0.27),(4.93±0.17),(6.02±0.86),and(6.43±0.66)h, and the differences between control group and experimental group were significant(P<0.05).The colony number of HepG2and QGY-7703 cells in control group and experimental group were 264.00 ± 12.62,269.00 ± 4.55, 165.00± 10.61,and 215.00 ± 4.43,and the differences between control group and experimental group were significant(P<0.01).Conclusion:HOXA13 can increase the proliferation,enhance the clone formation,decrease the number of cells at G1phase and increase the number of cells at S phase in the hepatocellular carcinoma HepG2 and QGY-7703 cells;and it may used as a new molecular target for diagnosis and treatment of hepatocellular carcinoma.
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Objective To evaluate the application effect of"2+2"medicine-education cooperation talents training mode. Methods The nursing undergraduates of 2013 grade were divided into the experimental and control group. The"2+2"medicine-education cooperation talents training mode was adopted in the former and traditional training model was used in the latter. The final examination scores of the fifth and sixth semester, scores of graduation examination, scores of the Nurse Competence Scale were compared between the two groups. Results There was no difference of scores of theory examination between the two group except the Nursing Professional English whose scores were higher in the control group. The practical skills exam scores of Medical Nursing, Surgical Nursing, Paediatric Nursing, Nursing of Gynecology and Obstetrics, Emergency Nursing, Rehabilitation Nursing, Community Nursing in the experimental group were 89.32 ± 6.02, 89.46 ± 5.29, 88.06 ± 6.34, 89.32 ± 5.58, 83.99 ± 6.44, 82.58 ± 5.78, 83.56±6.12, the control group were 87.72±5.90, 87.85±6.32, 85.59±5.79, 87.45±5.65, 81.82±6.06, 80.34± 5.89, 81.28±5.42, and the differences were statistically significant (t=2.052-3.261, P<0.05). The total and practical skills scores of graduation examination in the experimental group were 82.86±4.92, 85.60±4.54, which were 81.07 ± 5.52, 84.07 ± 4.59 in the control group. There were significant differences between the two groups (t=2.060, 2.011, P<0.05). The total score of Nurse Competence Scale and scores of the four dimensions of Teaching-coaching, Managing situations, Ensuring quality, Work role were 74.95 ± 3.40, 76.38 ± 3.98, 75.49 ± 3.23, 75.42 ± 2.72, 77.49 ± 2.76 in the experimental group, the control group were 73.63 ± 4.39, 72.90 ± 4.23, 74.12 ± 4.19, 74.32 ± 3.36, 76.46 ± 3.24, respectively. The differences were statistically significant (t=2.021- 2.492, P<0.05). Conclusions The"2 + 2"medicine- education cooperation talents training mode is proved to be an effective way to train nursing applied talents to meet regional demands. It could improve the hospital′s synthetic strength including health care quality and teaching ability, promote the development and construction of the nursing specialty and train the double-certificate teachers.
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OBJECTIVE@#To explore the therapeutic effect of Fasudil-modified splenic mononuclear cells (MNCs) in experimental autoimmune encephalomyelitis (EAE) and the possible mechanisms. @*METHODS@#C57BL/6 female mice were immunized with myelin oligodendrocyte glycoprotein peptide 35-55 to establish active immunity EAE model. Splenic MNCs were isolated on the 9th day after immunization and treated with or without Fasudil for 72 h in vitro. These cells were collected for analysis of the variance of T cell subtypes, the level of cytokines and the activity of Rho kinase (ROCK). MNCs (5×107 cells) were resuspended in 500 µL of phosphate buffer solution (PBS) and transferred into EAE model (intraperitoneal injection), which was divided into a PBS-MNCs group and a Fasudil-MNCs group. Changes of body weight and clinical symptom scores were observed. @*RESULTS@#Splenic encephalitogenic MNCs from EAE mice on the 9th day after immunization could establish passive transfer EAE model. But Fasudil-treated MNCs did not trigger EAE development. Compared with the PBS-MNCs group, the loss of body weight was less in the Fasudil-MNCs group. The in vitro experiment indicated that Fasudil could suppress the activity of ROCK on MNCs (P<0.01), decrease the percentage of CD4+ T cells with the expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) (IFN-γ: P<0.01; IL-17: P<0.05), while increase the secretion of CD4+ T cells with the expression of transforming growth factor-β (TGF-β) and IL-10 (all P<0.001) . Furthermore, Fasudil could inhibit the release of IL-17 (P<0.001) and enhance the level of IL-10 (P<0.05). @*CONCLUSION@#Fasudil-modified cell therapy affects the occurrence and development of EAE by inhibiting the inflammatory reaction of helper T cell 1 (Th1) and Th17 while enhancing the immunoregulative effect of Th2.
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Animals , Female , Mice , 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine , Encephalomyelitis, Autoimmune, Experimental , Interferon-gamma , Interleukin-10 , Interleukin-17 , Mice, Inbred C57BL , Myelin-Oligodendrocyte Glycoprotein , Spleen , T-Lymphocytes , Transforming Growth Factor beta , rho-Associated KinasesABSTRACT
Objective To investigate the immunoregulatory effect of Fasudil-modified macrophages on cell transferred experimental autoimmune encephalomyelitis ( EAE) in a mouse model.Methods Fe-male C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE.The encephalomyelitic mononuclear cells ( MNCs) were isolated from spleen of mice with EAE on day 9 after immunization and treated with or without Fasudil for 72 h in vitro.Several assays including the flow cytometry analysis, Griess reaction and ELISA were performed to analyze the M1 and M2 phenotypes of macrophages, the production of NO and the levels of cytokines, respectively.The cultured MNCs (5×107 cells) were resuspended in 500μl of PBS and transferred into na?ve C57BL/6 recipients via intraperitoneal injection.Two groups including the PBS-MNCs group and the Fasudil-MNCs group were set up.The body weights and clinical scores of the mice in each group were recorded in every other days after the induction of EAE in the recipients.Results The Fasudil treated MNCs affected the induction of EAE in adoptive cell transferred mice.The expression of CD16/32, iNOS and IL-12 on F4/80-macrophages were decreased, while the expression of CD206, CD23 and IL-10 on F4/80-macrophages were increased upon the treatment of Fasudil, indicating that Fasudil im-proved the differentiation of macrophages from M1 to M2 phenotypes.Moreover, Fasudil inhibited the pro-duction of NO and enhanced the expression of Arginase-1.Conclusion Fasudil ameliorated the clinical se-verity of EAE in mice by promoting the transformation of macrophages from M1 to M2 phenotype.
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BACKGROUND: Amnion has been widely used in ophthalmology. Numerous studies have suggested that amnion transplantation did not induce acute immunologic rejection. These indicated that amnion transplantation can be used as a safe material for repair of dural defects.OBJECTIVE: To study the probability of freeze-dried amniotic membrane (FDAM) as a dural substitue. METHODS: Each of the guinea pigs underwent bilateral parietal craniectomy behind the coronal suture and beside the midline to expose the dura. On the right side, a piece of dura mater was removed. The dural defect was covered with a piece of FDAM. The exposed dura on the left was cut and sutured itself as control. The animals in each group were sacrificed at 15, 30, 60 and 90 days after operation, respectively. The implants were harvested and stained with hematoxylin-eosin, and histologically analyzed. RESULTS AND CONCLUSION: After operation, the behavior of all guinea pigs remained completely normal. The wound healing was achieved in all cases. No wound infection, subcutaneous effusion or cerebrospinal fluid (CSF) leakage occurred. The graft was degraded gradually and covered with a sheet of connective tissue. Dural defects repaired with FDAM showed no adhesions to the brain surface. 15 days after operation, plenty of scattered fibroblasts appeared in the dural substitute. 30 days alter dural graft implantation, parts of the implant disappeared; meanwhile the hyperplasia of fibrous connective tissue took place in the center part of the dural substitute, without the infiltration of inflammatory cells. 60 days after implantation, a majority of the dural graft was degraded, substituted by fibrous connective tissue which was of hyperplasia and low-grade degeneration, surrounded by a small quantity of giant cells. 90 days after operation, colloidal degeneration happened in the dural substitute, surrounded by ossification tissue and the degenerated fibrous connective tissue. The inflammatory cells were not discovered. The animal experiment proves FDAM to be a safe and applicable dural substitute.
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Objective To study pharmacological mechanism of Sanshanxiao granules in treating diabetes. Methods The rat model of diabetes was set up by injecting STZ intraabdominally. The modeled diabetic animals were divided into model group, Sanshanxiao group, and Xiaokewan control group. Normal rats were devised as blank-contrasted group. It was observed through several aspects on the level of blood sugar and insulin, activity changes of SOD in serum, contents of MDA and changes of islets of pancreas morphologic contrastly. Results Sanshanxiao granules can reduce blood sugar, stimulate insulin secreted, develop activity of SOD and decrease the contents of MDA obviously (P
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Objective To explore the relationship between blood pressure changes and prognosis in patients with acute stroke by ambulatory blood pressure monitoring. Methods 79 patients with acute stroke were divided into two groups: cerebral infarction group (n=42) and cerebral hemorrhage group (n=37). Blood pressure of patients in both groups was monitored, and the score of handicap was evaluated respectively on 30d after onset. Results (1)Blood pressure was elevated in 87.3% of the patients with stroke at acute stage and then lowered spontaneously. Compared with that of the 1st day after onset, blood pressure lowered significantly from [SBP (155.9?21.7), DBP (96.6?20.8) mmHg] to [SBP (147.6?17.2), DBP (85.6?13.6) mmHg] (P