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Objective To discuss the clinical curative effect between dynamic external fixator (DEF) and hand orthosis (HO) in patients with middle phalanx pars basilaris fracture. Methods A total of 143 patients with middle phalanx pars basilaris fracture were selected from January 2015 to April 2016. The patients were divided into 2 groups according to the patients′select: DEF group (83 cases) and HO group (60 cases). The patients in DEF group were treated with DEF fixation under local nerve block anesthesia of digital root, and the patients in HO group were treated with HO fixation after manual repositioning. The fracture healing time and complications were observed. All patients were functionally trained and evaluated by total active motion (TAM). Results The patients were followed up for 8 to 16 weeks, with an average of 12 weeks. There was no statistical difference in fracture healing time between 2 groups (P＞0.05). All patients had postoperative pain, but the incidence of postoperative pain after removing DEF or HO in DEF group was significantly lower than that in HO group: 26.51% (22/83) vs. 46.67% (28/60), and there was statistical difference (P＜0.05). No redislocation or subluxation occurred after removal of external fixation in 2 groups. X-ray follow-up showed that the incidences of mild traumatic osteoarthritis, mild angulation deformity and mild lateral instability in DEF group were significantly lower than those in HO group: 6.02% (5/83) vs. 23.33% (14/60), 3.61% (3/83) vs. 33.33% (20/60) and 1.20% (1/83) vs. 26.67% (16/60), and there were statistical differences (P＜0.05). The excellent and good rate of TAM in DEF group was significantly higher than that in HO group: 83.13% (69/83) vs. 63.33% (38/60), and there was statistical difference (χ2=7.25, P＜0.01). Conclusions The clinical curative effect of DEF fixation is better than HO fixation in patients with middle phalanx pars basilaris fracture.
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Objective: To explore the clinical features and prognosis in patients with Takayasu arteritis (TA) combining neurological symptoms. Methods: We retrospectively studied 274 TA patients combining neurological symptoms who admitted to our hospital from 2002-01 to 2013-10 for their clinical and imaging features with prognosis. Results: The ratio of male to female was 1:4.3 and the mean age of disease onset was at (28.2±11.2) years. The most common neurological symptom was dizziness (214/274 cases, 78.1%), most frequent type was type III TA (112 cases, 40.9%), most common affected artery was left subclavian artery (147 cases, 53.6%), and there were 77 cases (28.1%) with (3-4) branches of the aortic arch involvement. For stroke conditions, ischemic stroke was more frequently observed in patients with steno-occlusive lesions in subclavian artery and common carotid artery, while hemorrhagic stroke was more frequently found in patients with steno-occlusive lesions in descending aorta, abdominal aorta and/or renal artery. Heart failure was the most common cause of death, it was also the most common cardiovascular event in surviving cohorts. Conclusion: TA patients could have many neurological symptoms, which were related to the number and site of artery involvement.
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Objective To explore the safety of the human bone marrow‐derived mesenchymal stem cells (hBMSCs) after silencing of human leukocyte antigen A2 expression .Methods We divided the cells into three groups:normal cultured cells of the 8th passage served as control group , and hBMSCs after HLA A2 silencing expression of the 5th and 15th passage as experimental groups 1 and 2 ,respectively .The hBMSCs were recultured by sterile methods .The growth curve ,telomerase activation ,and expressions of P27 ,cyclin D2 and cyclin‐dependent kinase 4 (CDK4) were utilized to explore the safety of the hBMSCs induced by LV‐siRNA‐HLA A2 .The BMSCs were transplanted to the subcutaneous layer of nude mice .Tissue types were detected 24 weeks after transplantation . Results The cell curves had no obvious left or right shift in all the groups . The telomerease activation in experimental groups 1 and 2 did not significantly differ from those in control group . The expressions of anti‐oncogene P27 ,cyclin D2 and CDK4 had no obvious difference between the two experimental groups and control group , either . There was only ectopic osteogenesis 24 weeks after the BMSCs (HLA A2 gene silenced ) were transplanted to the subcutaneous layer of the nude mice .Conclusion There was no obvious evidence to support that hBMSCs had undergone change in safety after the silencing of HLA A 2 expression .
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Objective: To explore a single center large cohort of patients with Takayasu’s arteritis for their clinical manifestation and long-term outcome in China. Methods: We retrospectively analyzed 566 patients with Takayasu’s arteritis admitted in our hospital from 2002-01 to 2013-11 for their clinical characteristics, laboratory ifndings, angiographic features, treatment and long-term outcomes. Results: The patient’s ratio for female to male gender was 1 to 3.8 and the average onset age was (28.9 ± 12.0) years. The most common non-speciifc symptom, initial symptom and complication were fever (52/566 patients, 9.2%), dizziness (214 patients, 37.8%) and hypertension (392 patients, 69.3%) respectively. The patients with pulmonary artery and coronary artery involvement were 83 (14.7%) and 66 (11.7%) respectively, and 131 (23.1%) patients had faster erythrocyte sedimentation rate. The major vascular damage was steno-occlusive lesion and the most common involvement was left sub-clavian artery, which was observed in 278 (49.1%) patients. The treatments were mainly included in medication, interventional therapy, autologous blood vessel transplantation, artiifcial blood vessel transplantation and aortic valve replacement. There were 32 patients died during the mean follow-up period of (5.0 ± 0.2) years. Hypertension, complication and the progressive stage of disease were the major factors affecting prognosis in relevant patients (regression coefifcients: 4.664, 1.959 and 1.870 respectively, allP<0.05). Conclusion: Hypertension was the leading reason for patients’ hospital visit. Takayasu’s arteritis was closely related to cardiovascular disease, the early diagnosis and treatment were really important in clinical practice.
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BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants. OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2. METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis. RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.
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10.3969/j.issn.2095-4344.2013.25.010
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Objective To construct the recombinant rAAV2-hGAD65 vector and detect its function both in vitro and in vivo. Methods The cDNA of human glutamic acid decarboxylase 2 (hGAD65) gene, which was one of gamma-aminobutyric acid (GABA) synthetase, cloned by the method of RT-PCR, was subcloned into the adeno-associated virus (AAV) vector and formed the recombinant vector of AAV-hGAD65 (rAAV2-hGAD65). The recombinant vector was packaged by the AAV Helper-Free System and its titer was detected. The primary fibroblast, cultured from the rat lung, was transfected by the rAAV2-hGAD65. The expression of the hGAD65 in fibroblast was detected by immunohistochemical method and the level of GABA in the media was assayed by high performance liquid chromatograph (HPLC). In vivo, the hGAD65 was detected by immunohistochemical method in STN and the concentration of the GABA in the reticular part of substantia nigra (SNr) was assayed by HPLC after the rAAV2-hGAD65 was injected into the subthalamic nucleus (STN) by the stereotaxic method. Results The sequence of hGAD65 cDNA was in accordance with that in the Genebank. The amino acid sequence of hGAD65 had no mutation and the titer of rAAV2-hGAD65 was reached 4.5 ×10~(11) per milliliter. The efficiency of infection to the rat primary firoblasts was 80%, and the concentration of GABA in the media of fibroblasts was (45.66±6.07)nmol/L per liter. In vivo, hGAD65 could be detected in STN, and the concentrateion of the GABA in the SNr was increased from (5.66±1.07)nmol/g to (12.66±2.59)nmol/g.Conclusion The cDNA of hGAD65 was cloned by RT-PCR and the recombinant vector of rAAV2-hGAD65 was constructed. The AAV can infect the primary fibroblast in vitro and the hGAD65 can catalyse the glutamic acic to GABA. In vivo, the concentration of GABA in the SNr was heighten after the rAAV2-hGAD65 was injected into the STN.
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BACKGROUND:With increased age,bone marrow mesenchymal stem cells(BMSCs)are influenced with regard to quantity and quality,which will induce great damages to the donors.Many studies have focused on seeking substitute MSC source.In contrast it remains controversial whether umbilical cord blood contains MSCs.OBJECTIVE:To isolate MSCs from human umbilical cord blood,and to detect their biological properties.METHODS:Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells.DMEM medium containing fetal bovine serum,penicillin,streptomycin and L-glutamine was added.Following several adherences and purification,the floating cells were discarded.Thus,many adherent cells with a confluence were collected.When cells were 60% 80% confluent,cells were digested by trypsin for subculture.Cells at passages,1,5 and 9 were obtained and their morphological changes were observed.Cell surface antigens were measured using flow cytometry.Growth curves were drawn,and cell viability was determined utilizing MTT.RESULTS AND CONCLUSION:Isolated umbilical cord blood MSCs presented an even size,showing spindle or star-shape fibroblasts-like cells.Umbilical cord blood MSCs at 1,5,9 passages were greatly positive for CD29,CD105 and CD166,but weakly positive for CD34 and CD45.Following 5 days of incubation,cells entered logarithmic growth phase.The number was decreased at day 9.Population doubling time was(53.5±8.32)hours.Cells grew well.Cells at 1-7 passages showed similar viability(P ＞ 0.05).Till passage 9,cell proliferation viability was decreased,but no significant difference was determined(P ＞0.05).Results verified that MSCs can be successfully isolated from umbilical cord blood in vitro.Cells at passages 1-9 presented a good reproductive activity.
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Objective To analyze the clinical features of takayasu's arteritis in elderly patients for improving their general management.Methods Twenty-six patients,aged over 60 years,with takayasu's arteritis were enrolled.The clinical manifestations and medical records were collected in detail and analyzed retrospectively.Results The mean age of the patients in our study was 64.0±3.8 years,four males and twenty-two females.Of all patients,nine were accompanied by coronary heart disease,seven by primary hypertension,four by diabetes mellitus,two by arrhythmia and one by subacute infective endocarditis.The frequent clinical manifestations were hypertension (n = 21,81%),dizzy (n=12.46%),chest pain (n=9,35%),pulselessness or weak pulse (n=7,27%).The clinical classification of this group showed brachiocephalic artery type (n = 8,30%),abdominal aorta type (n=4,15%),extensive type (n=11,58%) and pulmonary artery type (n=3,12%).Ten patients had elevated ESR level,six had elevated CRP level and fourteen had elevated ASO level.Two patients with diabetes mellitus died of serious complications.Conclusions Takayasu's arteritis in the elderly is usually accompanied by other cardiovascular diseases and risk factors.It is important to enhance the comprehensive treatment of these patients.
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Objective To examine the effect and mechanism of sodium ferulate inhibiting P38 MAPK in macrophage on neurotoxicity which was mediated by fribrilar-amyloid-induced.Methods The isolated peritoneal macropohages were cultured.P38 MAPK protein in nuclear extracts was analyzed by Western blot and production of tumor necrosis factor-?(TNF-?) was detected by ELISA.For neuron-macrophage co-cultures,Microtubute-associated protein-2(MAP-2) expression was detected by immunocytochemical technique.The level of LDH in the medium was measured.Results The production of TNF-? and P38 MAPK protein in nuclear extracts increased significantly by incubation with A?25-35.LDH efflux in neuron-macrophage co-culture medium increased and MAP-2 expression decreased significantly(P
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AIM To investigate the neuroprotective effects and possible mechanisms of saponins from Anemarrhena asphodeloides Bge. (SAaB) on neuronal damage induced by amyloid β-protein fragments 25-35 (Aβ25-35). METHODS Cultured mouse peritoneal macrophages were stimulated with Aβ25-35 (20 μmol·L-1) for 0.5, 1, 2 and 6 h or preincubated with SAaB (10, 30 and 100 μmol·L-1)for 10 min or mitogen-activated protein kinase (MAPK) specific inhibitors (p38 MAPK inhibitor SB 203580 and MEK specific inhibitor PD98059) for 30 min prior to the addition of Aβ25-35(20 μmol·L-1). After stimulation with Aβ25-35 for the indicated times, total cellular extracts were prepared for Western blotting of extracellular signal-regulated kinase (ERK) and p38 MAPK. After stimulation with Aβ25-35 for 48 h, the supernatants of cultured macrophages were collected for quantification of tumor necrosis factor-α (TNF-α) and nitric oxide (NO) and protein expression of inducible nitric oxide synthase (iNOS) in macrophages was determined by immunocytochemical staining. To determine whether SAaB has protective effect against neuronal apoptosis mediated by Aβ25-35-induced macrophages activation, macrophages were stimulated with Aβ25-35 in the presence or absence of SAaB (10, 30 and 100 μmol·L-1) for 48 h and then the cell-free supernatant of Aβ25-35-stimulated macrophages was transferred to the culture of cerebellar granule neurons for 72 h. Neuronal apoptosis was quantitated by scoring the percentage of cells with apoptotic nuclear morphology after Hoechst 33258 staining. RESULTS Aβ25-35(20 μmol·L-1) significantly induced increase in phospho-ERK1/2 and phospho-p38 MAPK protein expression without affecting total protein levels and in the production of TNF-α and NO in cultured macrophages. Aβ25-35-induced increase of TNF-α production in macrophages involved activation of ERK1/2 signal pathway. Importantly, TNF-α and NO generated by cultured macrophages after Aβ25-35 stimulation may be responsible for the majority of the neuronal apoptosis. SAaB (30 and 100 μmol·L-1) significantly suppressed Aβ25-35-induced increase in phospho-ERK1/2 and phospho-p38 MAPK protein. In addition, SAaB (10, 30 and 100 μmol·L-1) also decreased the level of TNF-α and NO in supernatants of cultured macrophage and inhibited Aβ25-35-induced increase in iNOS protein expression of macrophages. Neuronal apoptosis mediated by Aβ25-35-induced macrophage activation was also significantly attenuated by treatment with SAaB (10, 30 and 100 μmol·L-1). CONCLUSION SAaB protects neurons against the neuronal cell death induced by Aβ25-35. The beneficial effects of SAaB may be related to the reduction of TNF-α and NO from activated macrophage induced by Aβ25-35.
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Objective To determine the effects of ?-lipoic acid on bone metabolism in lead-poisoned rats.Methods Totally 31 SD rats were randomly divided into two groups,8 rats in blank control group and 23 in lead group that received gastric lavage with lead acetate(230 mg/L).After 40 days,3 rats from each group were taken for the measurement of lead in the blood and bone.The rats in lead group were then randomly divided into four groups: lead control group and three lead groups with different concentrations of ?-lipoic acid [30,60 and 100 mg/(kg?d)],with 5 rats in each.At the end of the experiment(80 d),the rats were killed and the samples of whole blood,serum and bone were collected to detect osteocalcin content with radioimmunoassay and alkaline phosphatase expression with immunohistochemistry staining.Results ① Blood and bone lead contents in each ?-lipoic treatment group were significantly lower than those in lead control group(P
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AIM: To investigate effect of sodium ferulate on A?25-35-mediated signaling pathway. METHODS: The isolated peritoneal macrophages from mice were cultured. p38 MAPK protein kinase in nuclear extracts was analyzed by Western blotting. The concentration of TNF-? and NO in supernatant were measured by ELISA and Griess reaction technique. The expression of iNOS protein was detected by immunochemical technique. RESULTS: A?25-35 significantly increased the concentrations of TNF-? and NO in supernatant, expression of iNOS in macrophages and p38 MAPK protein kinase in nuclear extracts, which were blocked by sodium ferulate. CONCLUSION: Sodium ferulate inhibits p38 MAPK activation triggered by A?25-35.
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Objective To study the effects of amniotic epithelial cells conditioned medium on the differentiation of bone marrow stromal cells into neural cells. Methods Bone marrow stromal cells and amniotic epithelial cells were isolated and cultured in vitro,then the cell surface antigen was detected by flow cytometry and the expressions of nestin and ki67 were detected by immunofluorescence staining method.When the cells were co-cultured with amniotic epithelial cells conditioned medium,the morphological character of cells was observed by inverse phase-contrast microscope,and the expressions of NSE(neurone specific enolase),TH(tyrosine hydroxylase) and DAT(dopamine transporter) were detected by immunofluorescence staining method. Results Amniotic epithelial cells conditioned medium had obvious inductive effect on bone marrow stromal cell's neural differentiation.Conclusion The amniotic epithelial cells conditioned medium may have inductive effect neuron-like cell's differentiation and dopaminergic neuron-like cell's differentiation of bone marrow stromal cells in vitro.
