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Objective:To investigate the effect of pilose antler polypeptide combined with Schwann cells modified by glial cell line-derived neurotrophic factor (GDNF)gene on the proliferation of human bone marrow mesenchymal stem cells (BMSCs)in vitro .Methods:According to the conventional method,the bone marrow (10 mL)was extracted from the healthy volunteers and was inoculated into the culture flask.The primary cultured cells were completely fused.The BMSCs were harvested at the 3rd generation and the cells were adjusted to 5 × 106 mL-1 . 4 μL GDNF gene modified Schwann cells was added into GDNF group,4 μL (10 mg· L-1 )PAP combined with GDNF gene modified Schwann cells was added into combination group,and only same amount of medium was added into control group.The proliferative activities,cell nuclear antigen (PCNA)levels and apoptotic rates of BMSCs in various groups were detected by enzyme-linked immunosorbent assay,ELISA method and Annexin Ⅴ-FIFC/PI cell apoptosis detection kit,respectively.Results:After primary culture for 48 h,most of the cells adhered to the wall,and the morphology of the cells changed into polygonal shape and few of them showed spindle.The passaged cells showed spindle spindle,the cells were confirmed as BMSCs,and all of them were non-hematopoietic stem cells.Compared with control group,the proliferative activities and the PCNA level of the BMSCs in GDNF group and combination group were increased (P <0.05)and the apoptotic rates were decreased (P <0.05);compared with GDNF group,the proliferative activity and the PCNA level of the BMSCs in combination group were increased (P < 0.05 )and the apoptotic rate was decreased (P < 0.05 ).Conclusion:PAP combined with Schwann cells modified by GDNF gene can promote the proliferation of human BMSCs in vitro .
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Objective To determine the effect of different doses vitamin D supplementation to the change of bone metabolism in 6 months after Roux-en-Y gastric bypass ( RYGB) .Methods The change of body weight and body mass index ( BMI ) in 36 patients in 6 months after RYGB WAS analyzed.Then,the effect of low-dose (n=11,400 IU/day) and high-dose (n=12,1 600 IU/day) vitamin D to the serum calcium,25-hydroxyvitamin D and bone density were observed ,and 15 patients as control .Results In 6 months after RYGB,the body weight and BMI were decreased ,and the differences were statistical significance (P<0.05).The serum calcium,25-hydroxyvitamin D and bone density were increased in high-dose group,and the differences were statistical significance (P<0.05).Conclusion High-dose (1 600 IU/day) vitamin D supplementation is effective to the bone loss of patients undergoing RYGB .
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PBL (Problem-based learning) is a kind of teaching method, which is helpful for guiding students to solve practical and complex problems. The compiling of PBL teaching plans is the key to the implementation of PBL teaching. According to the teaching outline of internal medicine, we set the PBL teaching plan, formulated the teaching goal, collected the clinical real cases, finished the compiling of PBL cases of student version and teacher version, and preceded teaching practice accord-ing to the PBL cases. Improving and constructing the compiling framework of internal medicine PBL teaching plan laid the theoretical foundation for carrying out internal medicine PBL teaching.
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Objective To compare the effect of atorvastatin and rosuvastatin on myocardial injure and inflammatory factors in patients with unstable angina pectoris(UAP)undergoing percutaneous coronary intervention(PCI).Methods Ninety six UAP pa-tients undergoing PCI were enrolled in the study and divided into two groups,patients were given atorvastatin(20 mg/d)or rosuv-astatin(20 mg/d)besides conventional treatment for 1 week before PCI.Datas were collected before medication,PCI and Twenty-four hours after PCI,including CK-MB,cTnI,hs-CRP,TNF-α,IL-6 and IL-10.Results Twenty-four hours after surgery,the PCI, CK-MB,cTnI,hs-CRP,TNF-α,IL-6 of rosuvastatin group were lower than those of atorvastatin group(P <0.05),but IL-10 of ro-suvastatin group was higher than that of atorvastatin group(P <0.05).Conclusion Rosuvastatin (20 mg/d)is better than atorvas-tatin (20 mg/d)in the efficacy of UAP patients undergoing PCI,which can reduce the level of myocardial injure and inflammatory factors effectively.
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Objective To evaluate the clinical effect of the upper lip defect with lower cross-lip flap (Abbe-Estlander flap).Methods A total of 68 cases of upper lip defect underwent the reconstruction with cross-lip flaps.We applied Abbe flap in upper lip defect in the center,and Estlander flap in lateral upper lip defect.This method was a two-stage procedure:the first stage was to rotate flap 180 degrees into the upper lip defect and to suture with the created defect of the upper lip,with careful maintenance of blood supply from the pedicle; the second was to undertake the division of bridged pedicle and paid more attention to creation of the vermilion border.Results The flaps survived in all cases.Evaluation from 3 to 12 months after the operation showed that the shape of lips were obviously repaired with excellent aesthetic and functional results.Conclusions Abbe-Estlander flap could reconstruct anatomical features and function of the lip precisely.It seems that within certain limits (probably between one-third and one-half of total upper lip length),this flap appears to be the preferred method for upper lip reconstruction.
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BACKGROUND:Myocardial fibrosis following myocardial infarction is an important mechanism of ventricle reconstitution. However, there are few reports concerning effects of myocardial transplantation related to stern cells on this process. OBJECTIVE: To investigate the effects of auto-skeletal muscle satellite cells implanted into ischemic myocardium on myocardial fibrosis in rats with myocardial infarction and their mechanisms.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Third Research Room, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from July to September 2007. MATERIALS: A total of 45 Wistar rats, of both genders, weighing 150-200 g, were used in this study. Of them, 30 rats were used to establish models of myocardial infarction.METHODS: A total of 45 rats were assigned to 3 groups (n=15). Rats in the myocardial infarction group received ligation of the left anterior descending coronary artery to induce myocardial infarction. 2 weeks later, 0.2 mL serum-free M199 medium was infused into the juncture between infarct region and normal myocardium through multiple points. In the transplantation group, following model induction, 0.2 mL auto-skeletal muscle satellite cells in rats after 2-weeks in vitro culture were transplanted into the surrounding of infarct region. Rats in the sham operation group were not induced to create models, only injected with 0.2 mL saline in the heart anterior wall surrounding the left anterior descending branch through multiple points. MAIN OUTCOME MEASURES: Four weeks after injection, vascular endothelial growth factor mRNA and vascular endothelial growth factor protein expression in the ischemic myocardium was demonstrated. Capillary density changes in the ischemic myocardium were detected. Growth and proliferation of myocardial cells in the infarct region were observed using hematoxylin-eosin staining.RESULTS: Vascular endothelial growth factor mRNA and vascular endothelial growth factor protein expression was significantly decreased in the sham operation and myocardial infarction groups compared with the transplantation group at 4 weeks following satellite cell transplantation (P<0.01). Capillary density was greater in the myocardial infarction group compared with the sham operation group (P<0.05). Capillary density was significantly higher in the rat ischemic myocardium in the transplantation group compared with the sham operation and myocardial infarction groups (P<0.01). Hematoxylin-eosin staining demonstrated that myocardial morphology was normal in rats of the sham operation group, with clear structure, orderly myocardial fibrosis. There were no fibroblastaggregation and hyperplasia among myocardial fibrosis. Fibroblast hyperplasia and collagent formation were found in the rat myocardium in the myocardial infarction group, with disorderly myocardial structure. Myocardial cells with transverse striation and many nuclei were observed in the rat infarct region of the transplantation group, with orderly arrangement. Fibrous tissue was significantly less in the transplantation group compared with the myocardial infarction group.CONCLUSION: Satellite cells can proliferate and differentiate into striated muscle-like cells with flexible and systolic functions in the infarct region. Satellite cells secrete vascular endothelial growth factor and promote blood capillary hyperplasia in ischemic myocardium by autocrine and paracrine, which finally effectively inhibits fibrosis progress in the ischomic myocardium.
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AIM: To investigate the effect of insulin on vascular smooth muscle cells (VSMCs) proliferation and to evaluate the intracellular signaling pathways involved. METHODS; VSMCs separated from Sprague - Dawley rats were used in this study. The proliferation of VSMCs induced by insulin was assayed by [~3H ] - thymidin incorporation. The protein expression and activity of p -ERK1/2 were determined by immunblot and [~(γ-32)P]ATP incorporation. RESULTS: Insulin induced cell proliferation in a concentration - dependent manner. The proliferative effect of insulin on VSMCs was inhibited partly by LY294002 (48.8% ) , an inhibitor of PI -3 kinase, and the ERK1/2 inhibitor PD98059 (43.6% ) , respectively. Moreover, phosphorylation of ERK1/2 and activity of ERK1/2 induced by insulin were also inhibited partly by LY294002.CONCLUSION: PI -3 kinase and ERK1/2 are involved in insulin induced VSMCs proliferation.
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[Objective] To study effects of the velvet antler polypeptides on spinal nerve cell apoptosis induced by ?-amyloid peptide. [Method]Spinal nerve cells were performed serial subcultivation,and entered into experiment during the exponential phase of growth.The viability of spinal nervecells 24h after induction by ?-amyloid peptide with different concentrations were detected by MTT assay,percentages of the cell apoptosis and expression of casepase-3 were detected after induction by 25?mol/L ?-amyloid peptide.[Result]It was revealed that 24 h after treatment with ?-amyloid peptide of different concentrations,the viability of spinal nerve cells was significantly decreased in a dose-dependent manner(P
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Background and objectives Proliferation of human vascular smooth muscle cells (VSMCs) induced by hyperinsulinemia is a very common clinical pathology. Extensive research has focused on PKC (Protein kinase C)-MAPK (mitogen-activated protein kinase)intracellular signal transduction and the phenotypic modulation accompanied by reorganization of intracellular F-actins in VSMCs.Methods DNA synthesis, signaling of ERK1/2 MAPKs, and changes in α-smooth muscle (SM) actin and F-actin were studied in hypertensive and normotensive human arterial VSMCs exposed to insulin and PMA with and without the PKC inhibitor, GF109203X.Results Differences among cell types in MAPK signaling, α-SM actin, and F-actin isoforms in VSMCs harvested from the arteries of patients with essential hypertension (EH) and normotension (NT) were identified in response to insulin treatment. Proliferation and activation of MAPK were more pronounced in EH VSMCs than in NEH VSMCs. Insulin exposure decreased expression of α-SM actin and was accompanied by rearrangement of intracellular F-actins in VSMCs, especially in the EH group. These effects were reversed by treatment with the PKC inhibitor. Conclusions Human mesenteric VSMCs of EH and NT patients differed in proliferation, MAPK signaling, and degree of changes in α-SM actin and F-actin isoforms immediately following insulin exposure in vitro.
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Objective To investigate the effect of auto-skeletal muscle satellite cell implantation into ischemic myocardium on cardiac function and the mechanisms.Methods Approximately 10 7 to 10 9 muscle satellite cells(SCs)were cultivated in vitro.The left anterior descending(LAD)artery was ligated in Wistar rats to create myocardial infarction(MI).Some rats only underwent sham operation served as control.Two weeks after MI,autologous SCs,serum-free culture medium and sodium chloride injection were injected into ischemic myocardium of implantation rats(n=15),control rats(n=15)and myocardium around LAD of sham operation rats(SO,n=15),respectively.Four weeks after injection,hemodynamic parameters and cardiac function in all groups were evaluated by polygraph system,capillary density in the ischemic myocardium was demonstrated by immunohistochemical method,serum VEGF concentration was examined by ELISA,and the differentiated myofibers from SCs in the infarcted site were observed by pathologic examination and immunohistochemical method.Results Four weeks after injection,the SCs had progressively differentiated into striated muscle fibers in the myocardial infarction site,and immunohistochemical analysis confirmed their skeletal muscle origin.Compared with the SO rats,systolic blood pressure(SBP),diastolic blood pressure(DBP),mean ar-tery pressure(MAP),left ventricular systolic pressure(LVSP)and dp/dtmaxwere markedly decreased(P
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Objective To explore the relationship of the polymorphism of E-selectin gene A561C and essential hypertension (EH) among Chinese people. Methods Genotypes of E-selectin were analyzed in 95 EH patients with age ≤70 and 101 normal controls people matched in age and gender by polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP). Serum levels of lipid, glucose, urea and creatinine were measured by biochemical techniques. Plasma soluble E-selectin were measured by enzyme-linked immunosorbent assay(ELISA). Results The frequency of E-selection genotypes AA, AC and CC in EH group were significantly higher than normal group (P
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Objective To evaluate the effects on blood pressure and vascular endothelial-1 function of combined therapy of atrovastin and nifedipine controlled released tablet in essential hypertension.Methods A randomized,double-blind,controlled trial was performed.Eighty-two patients with essential hypertension were randomly divided into two groups,receiving nifedipine controlled released tablet 30 mg once daily,or atrovastin 10 mg once daily and nifedipine controlled released tablet 30 mg once daily for 12 weeks.Another 30 subjects with normal blood pressure served as normal blood pressure control.Antihypertensive efficiency was observed during the whole study period.The concentration of plasma endothelin-1(ET-1) nitric oxide(NO) were measured before and after treatment.Results Blood pressure decreased significantly in both groups,but more significant in combined therapy group.In the combined therapy group,atrovastin resulted in a significant reduction of plasma ET-1 and a rise of nitric oxide(NO)(P
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<p><b>OBJECTIVE</b>To observe the effect of basic fibroblast growth factor (bFGF) on neovascularization and prefabricated flap survival.</p><p><b>METHODS</b>Male New-Zealand rabbits weighting 2.0-2.5 kg were used in this study. The experimental model used a prefabricated neck flap, supplied by the transferred and implanted central vascular bundle of the ear. 9 micrograms bFGF and 0.2 ml of normal saline was instilled in the vascular pedicle of the experimental and the control group respectively. After 1, 2, 3 weeks of operation, the neovascularization was studied by confocal laser scanning microscope (CLSM) and flap survival was observed.</p><p><b>RESULTS</b>The neovascularization and survival of the prefabricated flap was different in the experimental and the control groups. The experimental group was better than the control group.</p><p><b>CONCLUSION</b>bFGF treatment improved sprouting of the implanted vessel and the prefabricated flap survival.</p>
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Animals , Male , Rabbits , Fibroblast Growth Factor 2 , Pharmacology , Graft Survival , Physiology , Microscopy, Confocal , Neovascularization, Physiologic , Surgical Flaps , Physiology , Time FactorsABSTRACT
AIM:To explose the possible existing pathway of intracellular signaling transduction in hypertensive induced by insulin in rat vascular smooth muscle cells proliferation which involved mitogen-activated protein kinase. METHODS:Male spontaneously hypertensive rat (SHR) aorta and WKY(6 weeks old) were isolated and then cultured to make the purified vascular smooth muscle cells.6-8th generation of VSMC were interfered with insulin in vitro . MAPK activity was determined by myelin basic protein method and its volume was measured with Western Blot. And [ 3 H]-TdR was used to measure DNA synthesis in VSMC proliferation. RESULTS: After the interfered with insulin the DNA synthesis was increased obviously in SHR group. MAPK activity and its contains in SHR were increased more than the control group. Protein kinase C inhibitor decreased MAPK activity induced by insulin. CONCLUSION:Proliferation of SHR VSMC in vitro was correlated with increased activity of MAPK. Insulin can affect MAPK induced activity. So an insulin-PKC-MAPK axis may exist in hypertensive VSMC.
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AIM: To explore the effect of endoplasmic reticulum stress on cardiac myocyte apoptosis in mouse congestive heart failure induced by myocardial infraction.METHODS: The mouse model of heart failure was established by ligating the left anterior descending coronary to produce acute myocardial infarction.Thirty-two mice were divided into 4 groups: sham group and groups of post-operation at time points of 2,4 or 6 weeks,respectively.The ventricular dilatation and left ventricular functions were assessed by echocardiography.The expression of GRP78,CHOP,caspase-12,cleaved caspase-12,JNK and phosphorylated-JNK was detected by Western blotting.The cardiac myocyte apoptosis was determined by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL).RESULTS: The cardiac expression of endoplasmic reticulum chaperones GRP78 was significantly increased in the hearts with functional failure.The upregulated expression of CHOP,phosphorylated-JNK and cleaved caspase-12 illuminated that the CHOP-JNK-caspase-12 dependent pathways for endoplasmic reticulum-initiated apoptosis were activated in the heart with functional failure by myocardial infraction.CONCLUSION: These findings suggest that the congestive heart failure induced by myocardial infraction is associated with endoplasmic reticulum stress and activation of CHOP,JNK,caspase-12 dependent pathways for endoplasmic reticulum-initiated apoptosis.
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AIM: To identify proteins involved in insulin stimulation and the molecular mechanism of proliferation and migration in vascular smooth muscle cells (VSMCs). METHODS: A series of methods, including 2-D electrophoresis, PDQuest software analysis of 2-DE gels, peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and SWISS-PROT database searching, were used to separate and identify the differentially expressed proteins. The difference of some proteins was proved by Western blotting. Proliferation and migration of VSMCs treated with insulin were also observed. RESULTS: DNA synthesis were increased in VSMCs. ~3H-thymidine incorporation in VSMCs from SHR (14.40?0.85) was higher than that in VSMCs from WKY (9.21?0.93, P
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AIM: To investigate the changes of intracellular calcium ion (Ca2+) concentration in mouse H9c2 (2-1) cells transfected with or without FK506 binding protein 12.6(FKBP12.6) gene by ultrasound mediated destruction of microbubbles. METHODS: The pcDNA3.1-FKBP12.6 plasmid, mingled with albumin-coated microbubbles agents, was transfected into H9c2 (2-1) cells by ultrasound-mediated destruction of microbubbles. The H9c2 (2-1) cell growth state was investigated by inverted microscope. The changes of intracellular Ca2+ concentration was determined by laser scanning confocal microscope. The FKBP12.6 protein expression was checked by immunohistochemistry. RESULTS: As compared with control cells, the H9c2 (2-1) cells, transfected with FKBP12.6 gene, grew better, had higher gross intracellular Ca2+ concentration. CONCLUSION: FKBP12.6 gene augments Ca2+ concentration in mouse H9c2 (2-1) cells, enhances the contractibility of the myocardial cell, which may be helpful to improve the myocardial dysfunction.