Objective To observe the effects of EPO on GDNF secretion and cell cycle of Schwann cells(SCs) in vitro,and explore how EPO improve peripheral nerve regeneration. Methods Devided the purified Schwann cells of primary culture into three groups:group A with DMEM culture solution contained 10%fetal bovine serum and without EPO,group B with DMEM culture solution as above and 10 U/ml EPO,group C with DMEM culture solution as above and 50 U/ml EPO.The GDNF level of each group was detected in the culture by ELISA,and carried out the flow cytometry of Schwann cells on each group.Results GDNF in culture solution of group B,group C was more than that of group A,and the S-stage rate(S%)and (S+G_2M)%of Schwann cells in group B and group C more than that of group A. Conclusion EPO can increase the GDNF secretion and enhance proliferation activity of Schwann cells,which can explain how EPO improve peripheral nerve regeneration.

Objective To establish the separation and expansion method of human umbilical cord derived mesenchymal stem cells (hUCMSCs) and explore the influence of 1,25 (OH)2VD3 on hUCMSCs in vitro.Methods UCMSCs were cultured from adherent tissue pieces and detected by immunohistochemistry.Taking the third generation with good growth state UCMSCs make intervene experiment.Set 10-7 mol/L,10-9mol/L and 10-11 mol/L 3 kind of concentration of 1,25(OH)2VD3 as the experimental group,and a control group (DMEM medium).The morphologic change of UCMSCs was observed by inverted microscope.Observe the cell proliferation by the method of MTT.Detect alkaline phosphatase assay and type Ⅰ collagen assay from immunohistochemistry.And the calcium tubercle formation would be examined after 21 days.Results UCMSCs were cultured from adherent tissue pieces and strongly positive for CD44,CD105 and negative for CD34,CD45.consistent with the hUCMSCs characteristics.Under different concentration of 1,25 (OH)2VD3hUCMSCs proliferation were promoted within 7 days but were suppressed beyond 7 days.The high doses group ( 10-7 mol/L group ) obvious inhibitted the stem cell proliferation.Different concentrations of 1,25 (OH)2VD3 and days have interactive effect (P ＜ 0.05).The differences between different groups with ALP and type Ⅰ collagen assay has statistical difference (P ＜ 0.05).The secretion of ALP and type Ⅰ collagen assay of 10-7 mol/Lconcentration group was higher than others.Typical mineralization nedus was found in intervene groups.Conclusion HUCMSCs can be successfully cultured from the adherent tissue pieces.1,25(OH)2VD3 can effectively induce hUCMSCs to differentiate into osteoblasts in vitro.

Objective To study the biological effect of Icariine on human umbilical cord derived mesenchymal stem cells(huCMSCs) and inducing them to differentiate into osteoblasts. Methods MSCs were isolated and expanded in vitro and their surface antigens of huCMSCs were detected by immunohistochemistry. HuCMSCs were assigned into two groups. In the blank control group, huMSCs were incubated in basic medium. In the experimental group huMSCs were incubated in Icariine medium, The proliferation and differentiation of huMSCs were examined by inverted microscope, Alkaline phosphatase (ALP) staining and the determination of ALP activities. Results HuCMSCs were strongly positive for CD44, and negative for CD34; strongly positive expression for ALP and calcium deposition was detected from the second day to the fourth day in Icariine group; blank control group and Icariine group had significant differences (P ＜ 0.05).Conclusion HuCMSCs can be successfully cultured from the adherent tissue pieces, Icariine enhances proliferation and osteogenic differentiation of huCMSCs and be used as an osteoinductive factor.

30 kg/m2 served as obesity group.Using Gross formula,the true total blood loss was calculated depending on the height,body mass and pre-and post-operative Hct,and hidden blood loss was gotten by subtracting the visible blood loss from total loss.③The correlation between visible and hidden blood loss was observed.RESULTS:All 41 patients undergoing THA and 37 TKA were involved in the result analysis.①Following THA,the mean total loss was 1 520 mL and the hidden blood loss 482 mL(32%).Following TKA,the mean total loss was 1 508 mL and the hidden blood loss was 776 mL(52%).The hidden blood loss between THA and TKA was significantly different(P 0.05).CONCLUSION:①The hidden blood loss in TKA is far higher than THA.It is very important to supplement the blood volume during TKA for the insufficiency to regain general circulation although using blood re-infusion.②It may improve clinical evaluation capabilities to estimate hidden blood loss and thus result in better patient care after joint arthroplasty.

Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.

Objective To investigate the biological characteristics of human umbilical cord derived mesenchymal stem cells(UCMSCs) and inducing them to differentiate into neurons in vitro,in order to provide stem cell resource for the tissue engineering artificial nerve. Methods UCMSCs were cultured from Wharton jelly of human umbilical cord in the condition of sterilitas,surface antigens of UCMSCs were detected by immunohistochemistry.The frist group of cells were added by virgin nutrient,the second group of cells were induced by merely growth factors,the third of cells were induced by merely astragalus mongholicus and the fourth by the astragalus mongholicus with growth factor,observed the morphology of the cells of the two groups under inverted microscope,and determine the positive expression rate of nerve cells' markers,such as NSE,NF and GFAP.The results were statistically analyzed.Results UCMSCs were strongly positive for CD29,CD44,CD105,weakly positive for CD105 and negative for CD34,CD45.After induced by Astragalus mongholicus,UCMSCs were strongly positive for GFAP,NSE. Conclusion UCMSCs can be successfully cultured from the adherent tissue pieces,Astragalus mongholicus can successfully induce them to differentiate into neurons in vitro,to provide a new method of making the tissue engineering artificial nerve and treat theperipheraly nervus defect.

Objective:To investigate the effects of curcumin on apoptosis and radiosensitivity of human chordoma cell line CM-319 and its mechanism.Methods:CM-319 cells were cultured in vitro and divided into curcumin 0 μmol/L group, curcumin 5 μmol/L group, curcumin 10 μmol/L group, curcumin 20 μmol/L group, si-NC group, si-UHRF1 group, curcumin+pcDNA group, curcumin+pcDNA-UHRF1 group.Flow cytometry was usedtodetect cell apoptosis, Western Blot was used to detectthe protein expression of Bcl-2, Bax, Cleaved Caspase-3 and UHRF1, qRT-PCR was used to detect the expression of UHRF1 mRNA, and clone formationexperiment was used to detect the sensitivity of CM-319 cells to radiation.Results:Compared with the curcumin 0 μmol/L group, the apoptosis rate of CM-319 cellsinthe curcumin 5 μmol/L group, curcumin 10 μmol/L group and curcumin 20 μmol/L groupincreased 12.61%±1.57%, 18.42%±1.54%, 29.37%±2.65% vs 7.24%±0.73% ( t=9.305, 19.680, 24.153; all P<0.05), and the protein expression of Bcl-2 decreased 0.62±0.06, 0.51±0.05, 0.33±0.03 vs 0.76±0.07 ( t=4.556, 8.719, 16.939; all P<0.05), while the protein expression of Bax 0.41±0.04, 0.55±0.05, 0.69±0.06 vs 0.26±0.03 ( t=9.000, 14.920, 19.230; all P<0.05) and Cleaved Caspase-3 0.37±0.04, 0.49±0.05, 0.62±0.06 vs 0.24±0.03 ( t=7.800, 12.862, 16.994; all P<0.05) increased, and the expression of UHRF1 mRNA decreased 0.82±0.08, 0.67±0.07, 0.42±0.04 vs 1.01±0.09 ( t=4.734, 8.946, 17.972; all P<0.05), the protein expression of UHRF1 decreased 0.61±0.06, 0.48±0.05, 0.31±0.03 vs 0.83±0.08 ( t=6.600, 11.130, 18.258; all P<0.05), and were dose-dependent, and the sensitization ratios were 1.433, 1.708, and 2.183, respectively. Compared with the si-NC group, the protein expression of UHRF1in CM-319 cells in the si-UHRF1 group decreased 0.35±0.04 vs 0.79±0.08 ( t=14.758, P<0.05), the apoptosis rate of CM-319 cells increased 21.49%±2.11% vs 8.24%±0.83% ( t=17.531, P<0.05), the protein expression of Bcl-2 decreased 0.34±0.03 vs 0.71±0.07 ( t=14.575, P<0.05), while the protein expression of Bax 0.62±0.06 vs 0.25±0.03 ( t=16.547, P<0.05) and Cleaved Caspase-3 0.58±0.05 vs 0.23±0.03 ( t=18.007, P<0.05) increased, and the sensitization ratio was 1.727. Compared with the curcumin+pcDNA group, the apoptosis rate of CM-319 cells in the curcumin+pcDNA-UHRF1 groupreduced 13.59%±1.25% vs 28.31%±3.01% ( t=13.549, P<0.05), the protein expression of Bcl-2 increased 0.63±0.06 vs 0.31±0.03 ( t=14.311, P<0.05), while the protein expression of Bax 0.42±0.04 vs 0.71±0.07 ( t=10.791, P<0.05) and Cleaved Caspase-3 0.38±0.04 vs 0.65±0.05 ( t=12.650, P<0.05) decreased, and the sensitization ratio was 0.539. Conclusion:Curcumin can induce apoptosis of CM-319 cells and enhance the radiosensitivity of CM-319 cells, , and the mechanism is related to the down-regulation of UHRF1 expression.

Objective To evaluate the clinical efficacy of local intra-arterial thrombolysis (LIT) with urokinase in patients with acute ischemic stroke.Methods One hundred and sixty two patients with acute ischemic stroke were treated with LIT by using urokinase and relationship of reconalization for different occluded arteries with the Glasgow outcome scale (GOS) scores three months later was analyzed. Results Angiography showed occlusion of the cerebral artery in 162 patients, among which 119(73.5%) patients showed the sites in the internal carotid artery system, with 27 occlusions in the internal carotid artery (ICA)trunk, 63(38.89%) in the middle cerebral artery (MCA)and 29(17.9%) in the anterior cerebral artery (ACA), and the remaining 43(26.5%)patients of vertebrobasilar artery (VBA); successful recanalization was achieved in 103 (63.58%)patients, including 11 (40.7%), 49(77.8%), 20(69.0%) and 23(53.3%), respectively, after intraarterial infusion of urokinase. Unsuccessful recanalization occurred in 59patients (36.42%). Followed up for 90 days, 90 (55.6%) patients obtained a good outcome; 72(44.44%) had poor prognosis including 20(12.35%) deaths. 8 patients associated with hemorrhage (4.9%) 73 with reperfusion injury (45.1%) and 5 arterial re-occlusion (3.1%). Based on statistic analysis, ICA trunk and VBA had low ratio of successful recanalization with poor clinic prognosis MCA and ACA possessed high ratio of successful recanalization and good clinic outcomes. There was a significant relationship between arterial recanalization rate and clinic prognosis (r=0.86).Conclusions Successful recanalization of cerebral occlusive artery by using intra-arterial thrombolysis could improve clinic prognosis in patients with acute ischemic stroke. Clinical prognosis has a significant relationship with both initial treatment time and arterial recanalization rate.

Objective To explore the causes and preventive measures of symptomatic intracranial hemorrhage in 217 patients with acute cerebral ischemic stroke treated with local intra-arterial urokinase. Methods From February1999 to June 2004, 217 patients were treated for acute ischemic stroke with local intra-arterial urokinase in our hospital. Factors associated with symptomatic intracranial hemorrhage of intra-arterial thrombolysis were analyzed by Stepwise logistic regression to identify some factors relating the prediction symptomatic intracranial hemorrhage. Results Symptomatic intracranial hemorrhage occurred in 8 cases (3.7%). Predictors of the symptomatic intracranial hemorrhage were the elevated systolic blood pressure before therapy (odds ratio, 1.096; 95% CI, 1.006 to 1.194) and urokinase (UK) treatment (odds ratio , 1.068; 95% CL, 1.053 to 1.247). Risk of secondary symptomatic intracranial hemorrhage was increased with elevated systolic blood pressure. Other factors like age, initial treating time, NIHSS, diabetes and collateral circulation did not predict the symptomatic intracranial hemorrhage respectively. Conclusions Predictors of symptomatic intracranial hemorrhage after local intra-arterial infusion of urokinase for acute ischemic stroke were the elevated systolic blood pressure before therapy and urokinase (UK) treatment.

Objective To study the chemical constituents in the leaves of Mallotus apelta. Methods Constituents isolation and purification were carried out on silica gel and polyamide column. Their structures were identified by physicochemical properties and spectral analysis. Results Five compounds were isolated and elucidated as taraxerol (Ⅰ), ?-sitosterol (Ⅱ), 5, 7-dihydroxy-6-prenyl-4′-methoxy-flavanone (Ⅲ), apigenin (Ⅳ), and apigenin-7-O[WTBZ]-?-D-glucoside (Ⅴ). Conclusion Compound Ⅲ is a new compound named as mallotusin. Compounds Ⅰ and Ⅲ-Ⅴ are isolated from the leaves of M. apelta for the first time.