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Objective To observe the proliferation ability of cocultured dendritic cells(DCs)loaded with tumor antigen and cytokine-induced killer cells( CIKs)and its killing effect on hepatocarcinoma cells HepG2. Methods The antigen of hepatocarcinoma cells HepG2 was prepared by repeated freezing and thawing of liquid nitrogen. Peripheral blood mononuclear cells(PBMNCs)were isolated from healthy donors by blood cell separator,then DCs and CIKs were induced. Ag-DCs were obtained by impinging DCs with tumor antigens. CIKs were divided into three groups:the first group was CIKs alone,the second group was mixed in the propor-tion of DCs : CIKs = 1 : 5,and the third group was mixed in the proportion of Ag-DCs : CIKs = 1 : 5. The three groups of cells were recorded as CIK group,DC-CIK group and Ag-DC-CIK group. The proliferation and cell phenotype of the three groups of cells were observed and the killing effects on hepatocarcinoma cells HepG2 were detected by methyl thiazolyl tetrazolium(MTT)assay. Results The proliferation multiples of the three groups of cells were gradually increased with the prolongation of culture time,and the proliferation rates of Ag-DC-CIK on the 9th day(61. 32 ± 1. 72),the 12th day(190. 83 ± 3. 53)and the 15th day(399. 09 ± 5. 60) were significantly higher than those of CIK group(22. 47 ± 2. 07,55. 91 ± 1. 81,83. 20 ± 2. 34)and DC-CIK group(40. 26 ±2. 49,125. 03 ±4. 16,251. 55 ±3. 25),and the difference between the three group was statisti-cally significant(F =185. 78,P =0. 033;F = 297. 35,P = 0. 018;F = 455. 37,P < 0. 001),in addition,the differences between each two groups were statistically significant(all P <0. 05). The cytotoxicity of Ag-DC-CIK to HepG2 cells at the effective target ratios of 5 : 1(31. 71％ ±0. 29％ ),10 : 1(42. 43％ ±1. 86％ )and 20 : 1 (57. 69％ ±1. 11％ )were significantly higher than those of CIK group(12. 11％ ±1. 14％ ,21. 30％ ±0. 52％ , 30. 71％ ±1. 26％ )and DC-CIK group(20. 06％ ± 0. 67％ ,29. 89％ ± 1. 37％ ,39. 11％ ± 0. 92％ ),and the difference between the three group was statistically significant(F =159. 64,P =0. 037;F =199. 36,P =0. 025;F =302. 08,P <0. 001),in addition,the differences between each two groups were statistically significant(all P <0. 05). On the 15th day of cell culture,the flow cytometry analysis showed that all the three groups were expressed CD3 + CD8 + ,CD3 + CD56 + double positive cells,the contents of CD3 + CD8 + 、CD3 + CD56 + double positive cells in the Ag-DC-CIK group(88. 12％ ± 1. 24％ ,61. 35％ ± 2. 63％ )were significantly higher than those in the CIK group(54. 37％ ± 3. 08％ ,18. 22％ ± 1. 83％ )and DC-CIK group(69. 80％ ± 1. 46％ , 39. 51％ ±2. 17％ ),and the difference between the three group was statistically significant(F = 414. 32,P <0. 001;F =378. 60,P <0. 001),in addition,the differences between each two groups were statistically signifi-cant(all P <0. 001). Conclusion The proliferation ability and killing effect of Ag-DC-CIK that obtained from antigen-pulsed DCs co-cultured with CIKs are significantly higher than those of CIKs and DC-CIKs.
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The views from teachers and residents were analyzed to discusses the problems existing in the standardized training of TCM residents,and try to find the solution.A total of 98 standardized training residents and 111 teachers from Dongfang Hospital affiliated to Beijing University of Chinese Medicine (BUCM) were enrolled.The questionnaire sceened by star APP on the phone was used to investigate them with six practical questions,each of which had three options.Both groups agreed that undergraduates' rotate period for 3 years,masters' period for 2 years,and doctors' period for 1 year.As to the item of TCM hospital selection,both groups chose traditional Chinese medicine hospital for resident standardization training.As to the item of majors,59 teachers (53.2％) and 67 (68.4％) residents chose to learn their majors.The teachers suggested that student independent management can improve sense of presence in the department.But 36.7％ of residents considered increasing income can improve sense of presence.There are some problems in TCM residents training system,and we suggest that it should be proceeding from reality in residents and constantly improved in the future.
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Up to 2015, almost all of the Third-grade class-A TCM hospitals in Beijing set up multiple sub-hospitals, under the regional integrating of Beijing-Tianjin-Hebei region, in order to meet regional homogenization of medical and health care demand, to provide high-quality medical service for the patients, and to improve the hospital development in a certain stage. Taking Beijing university of traditional Chinese medicine affiliated Dongfang hospital for example, it creates "two sub-hospitals " a court layout of mode of "one hospital with multiple branches". This article analyzed the sub-hospitals organization structure reform, cost budget control, performance management & personnel training, information system construction, to draw lessons from practical manage experience, and explored the "multiple sub-hospitals " superior management mode for our hospital.
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Up to 2015, almost all of the Third-grade class-A TCM hospitals in Beijing set up multiple sub-hospitals, under the regional integrating of Beijing-Tianjin-Hebei region, in order to meet regional homogenization of medical and health care demand, to provide high-quality medical service for the patients, and to improve the hospital development in a certain stage. Taking Beijing university of traditional Chinese medicine affiliated Dongfang hospital for example, it creates "two sub-hospitals " a court layout of mode of "one hospital with multiple branches". This article analyzed the sub-hospitals organization structure reform, cost budget control, performance management & personnel training, information system construction, to draw lessons from practical manage experience, and explored the "multiple sub-hospitals " superior management mode for our hospital.
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Objective To investigate whether hepatocyte growth factor ( HGF ) and insulin-like growth factor (IGF1) induce cardiac stem cells (CSCs) to proliferate and directly differentiate into cardiomyocytes in vitro.Methods The myocardial tissues were dissected for primary culture of CSCs with the method of explants .The expressions of c-kit and CD34 were examined with immunofluorescence .Primary cells were purified with c-kit by flow cytometry.CFDA SE fluorescent probe was used to detect the proliferation of c-kit+CSCs.C-kit +CSCs were divided into two groups , and cardiac stem cells group and co-cultured with cardiomyocytes group , both group were cultured with HGF and IGF 1.An inverted microscope was used to observe changes in cell number and morphology in different periods .Living cells workstation was used to observe CFDA SE fluorescence intensity , to acquire images and do statistical analysis .Immunofluorescence technique was used to detect the expression of Nkx 2.5 and cardiac troponin T .Results In cardiac stem cells group ,CSCs had no obvious changes in cell number .In co-cultured with cardiomyocytes group , CSCs proliferated and had changes in morphology .Nkx2.5 and cTnT were positively expressed . Several CSCs differentiated into beating cardiomyocytes . Conclusion In co-cultured with cardiomyocytes condition , HGF and IGF1 may promote CSCs to proliferate and differentiate into beating cardiomyocytes .
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BACKGROUND: Angiotensin Ⅱ (Ang Ⅱ) can induce cardiac hypertrophy and platelet-derived growth factor(PDGF) also stimulates cardiac hypertrophy. Is AngⅡ responsible for the pathogenesis of cardiac hypertrophy by inducing PDGF receptor expression?OBJECTIVE: To investigate the effect of mitogen activated protein kinase (MAPK) on the role of cardiac hypertrophy induced by Ang Ⅱ in cardiac myocytes so as to provide theoretical basis for clinical prevention and cure of cardiac hypertrophy.DESIGN: Controlled experimental study taking cardiac myocytes of cultured neonatal rats as subjects.SETTING: Department of pathophysiology in a university.MATERIALS: The experiment was completed in the Department of Pathophysiology, Medical College of Peking University. A total of 80 Wistar rats of either gender, aged 1 - 3 days, were provided by the Animal Center of Medical College, Peking University. Their hearts were removed for myocyte culture in the Cell Culture Laboratory.INTERVENTIONS: The cultured neonatal rat cardiac myocytes treated with 10-7mol/L Ang Ⅱ were Ang Ⅱ group, and those preincubated with 10-5mol/L PD98059(an antagonist of MAPK) for 30 minutes and then treated with Ang Ⅱ were PD98059 group. Cardiac myocytes of normal neonatal rats were as control group. The expression of PDGF-β was detected by western blot at 24 hours.MAIN OUTCOME MEASURES: Content of PDGF-β receptor in neonatal rat cardiac myocytes.RESULTS: The expression of PDGF-β receptor induced by Ang Ⅱ at neonatal rat cardiac myocytes markedly increased at 24 hours (432.41 ± 54.08) compared with that of control group(197.65 ± 44. 10) ( q = 6.77, P< 0.01 ). PDGF-β receptor expression of PD98059 group(317.2 ± 21.12) decreased compared with that of Ang Ⅱ group(q = 3.91, P < 0.05) .However, the expression did not return to the level of control group, and there was significant difference between PD98059 group and control group( q= 3.85, P <0.05).CONCLUSION: The results indicate that angiotensin Ⅱ promotes cardiac hypertrophy through inducing expression of PDGF receptor, in which mitogen activated protein kinase participates in. Maybe it is another important mechanism for Ang Ⅱ -induced cardiac hypertrophy. The results can provide experimental data for the primary and secondary prevention in heart rehabilitation.
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BACKGROUND: Neuropeptide Y (NPY) neurons are extensively located in various brain regions such as cerebral cortex, caudate-putamen nucleus, syslimbic system, thalamus and brain stem. They are also involved in various brain activities such as motor learning, motor memory and motor integration. Considering the fact that cerebellum can reorganize through motor learning, we tried to identify the morphology and distribution of NPY neurons in rat's cerebellar cortex to obtain the morphologic knowledge that is related to its cerebellar-cortex-based motor learning.OBJECTIVE: To investigate the morphology and distribution of NPY -immunoreactive neurons in rat's cerebellar cortex, and discuss the relationship between NPY neurons and cerebellum motor learning and motor memory.DESIGN: A single-sample-study based on animal samples.SETTING: Anatomy Department, Pathophysiology Department and Morphology Center in Xinxiang Medical College.MATERIALS: From July to December 2001, the experiment was performed at the Morphology Center in Xinxiang Medical College. Ten Sprague-Dawley (SD) rats, clean grade, regardless of their gender and weighing 100-200 g,were selected.METHODS: After intraperitoneal injection anesthesia and ascending aorta infusion fixation, the cerebellum was taken out by craniosurgery. The cerebellum was immersed in the same fixative fluid for duration of 48 hours, and then was embedded in paraffin. The next step was to make continuous sagittal sections. NPY neurons were identified by SP immunohistochemical staining, using rats cerebral section as the positive control. In the negative control, the first antibody replaced by Bovine Calf Serum(BCS), and the second antibody replaced by 0.01 mol/L PBS. Sequentially the light-microscopic observation and micrography were recorded.MAIN OUTCOME MEASUREMENTS: The Morphology and distribution of NPY neurons in rat's cerebellar cortex were taken as main outcome measurements.RESULTS: NPY-immunoreactive neurons were distributed in the Purkinje layer of the cerebellum. They had pear-like or spherical shapes, clear and unstained nucleus, unstained axons and dendrites, and their axons stretching out into the granular layer while the reticular dendrites elongated into the molecular layer. NPY-immunoreactive fibers, instead of neurons, were found in the molecular layer and granular layer.CONCLUSION: NPY-immunoreactive neurons in the Purkinje layer of the cerebellum may be involved in motor learning, motor memory, visceral activity and higher motor integration.
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AIM: To investigate the crosstalk between angiotensin Ⅱ (AngⅡ)-mediated and platelet-derived growth factor (PDGF)-mediated signal transduction in vascular smooth muscle proliferation.METHODS: A model of renal hypertension was made by two kidney/one-clip operation. Level of PDGF receptor β subunit of aorta was measured by Western Blot analysis. The effect of Ang Ⅱ on PDGF receptor β subunit expression was investigated in culture rat aortic vascular smooth muscle cells (VSMC).RESULTS: Systolic blood pressure obviously increased at 8th week after operation, whereas the level of PDGF receptor β subunit of aorta significantly increased by 126.6% (P＜0.05) in 2K1C rats compared with control group. The expression of PDGF receptor β subunit in cultured VSMC stimulated by AngⅡ was higher than that of control by 192.74%(P＜0.01). The effect of AngⅡ was inhibited remarkably by pretreated with losartan, a kind of specific AngⅡ receptor 1 (AT1) subtype antagonist and U73122, a kind of phospholipase C inhibitor. The effect was partly blocked by PD98059, which inhibit the activity of mitogen-activated, ERK-activating kinase (MEK).CONCLUSION: AngⅡ-induced PDGF receptor β subunit expression is regulated by the AT1 and its downstream signal molecule-PLC and ERK, might participate in the intracellular signal transduction pathway.
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AIM: To investigate the crosstalk between angiotensin Ⅱ (AngⅡ)-mediated and platelet-derived growth factor (PDGF)-mediated signal transduction in vascular smooth muscle proliferation.METHODS: A model of renal hypertension was made by two kidney/one-clip operation. Level of PDGF receptor ? subunit of aorta was measured by Western Blot analysis. The effect of Ang Ⅱ on PDGF receptor ? subunit expression was investigated in culture rat aortic vascular smooth muscle cells (VSMC).RESULTS: Systolic blood pressure obviously increased at 8th week after operation, whereas the level of PDGF receptor ? subunit of aorta significantly increased by 126.6% ( P
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AIM: To investigate the role of G?q/11-mediated signal transduction pathway in cardiac hypertrophy induced by angiotensin II (AngII). METHODS: Renal hypertension was performed by placing a sliver clip around the left renal artery(2K1C). Hemodynamic parameters, the ratio of left ventricular weight to body weight, the AngII contents and the activities of phospholipase C (PLC) in myocardium were measured at 1st, 2nd, 4th and 8th week after operation, respectively. The levels of G?q/11 were assayed by Western blot analysis. -leucine incorporation and levels of G?q/11 were measured in cultured neonatal rats ventricular myocyte after AngII stimulation. RESULTS: In 2K1C group, blood pressure and the ratio of left ventricle of heart to body weight were significantly increased compared with sham group between 2nd to 8th week. AngII content increased from 1st to 8th week after operation. Compared with the sham group, levels of G?q/11 in 2K1C group increased by 25.0% and 35.8% at 4th and 8th week( P