Objective To investigate the possibility、 efficacy、 safety of the repair of infarcted myocardium and improvement of cardiac function by intracoronary autologous mononuclear bone marrow cell (MBMC) transplantation in patients with AMI. Methods Thirty-eight patients with AMI were enrolled in this prospective study and divided into PCI+cell transplantation group and PCI only group. Baseline and follow up evaluations included complete clinical and laboratory evaluations, 2D echocardiogram, positron emission tomography (PET), 48 ECG monitoring. MBMC were harvested, isolated, washed, and resuspended in saline for infusion. 21 patients were transplanted with autologous MBMC via a balloon catheter placed into the infarct-related artery during balloon dilatation by highpressure infusion 14.6 days after AMI, which was performed 6-8 times for 2 minutes each and 7 patients were transplanted via non-selective infarct-related arteries by highpressure infusion. Results Surgery was safe. 3 patients had limited premature ventricular contractions during cell infusion for several seconds. 3 patients felt cold after 15-30 minutes infusing cell. After 3 months of fellow-up, there were viable myocardium in the infarct defected regions in 13 cases after MBMCs transplantation determined by PET, which occupied [(40.08?8.82%), P

Objective To investigate the possibility, effect and safety of intracoronary autologous moronuclear bone marrow cell (MBMC) transplantation in patients with ischemia heart failure (IHF). Methods 41 patients with IHF were enrolled in this prospective nonrandomized study. 14 patients were transplanted with autologous MBMC via a balloon catheter placed into the infarct-related artery during balloon dilatation by highpressure infusion, which was performed 6-8 times for 2 minutes each. 13 patients were transplanted via selective the infarct-related arteries by highpressure infusion. Results There were no major periprocedural complications. Two patients had limited premature ventricular contractions during cell infusion forseveral seconds. Two patients felt cold after 15-30 minutes infusing cell and got better several minutes later. There were no new onset of arrhythmias found on 48 h ECG monitoring. After 3 months of follow up, the symptoms and cardiac function were significantly improved in the transplantation group. FDG-PET analysis revealed a significant increase in myocardial metabolism (23.94?7.28)% (P=0.015). Plasma BNP lever decreased significatly at 3 days and 7 days after transplantation than before transplantation (P

[Objective]To evaluate the value of clinical examination and magnetic resonance imaging in diagnosing chronic anterior cruciate ligament injury.[Method]Sixty-five patients with the diagnosis of chronic anterior cruciate ligament injury were retrospectively analyzed about the course of diagnosis and treatment.To gain the primary diagnosis through clinical examination,8 patients were performed magnetic resonance imaging.Finally all the patients were carried out arthroscopic surgery to make a final diagnosis.[Result]Arthroscopy found 53 cases with complete anterior cruciate ligament tears,12 cases with partial anterior cruciate ligament tears.In the complete anterior cruciate ligament tears cases,79.2% patients had positive anterior drawer test,96.2% had positive Lachman test and 92.5 % had positive pivot shift test.In the partial anterior cruciate ligament tears cases,16.7% patients had positive anterior drawer test,50.0% had positive Lachman test and 33.3 % had positive pivot shift test.The accuracy of MRI in diagnosing anterior cruciate ligament tears was 100%.[Conclusion]Clinical examination and magnetic resonance imaging can diagnose chronic anterior cruciate ligament effectively.

Objective To explore the role of apelin-13 in regulating stem cell differentiation into vascular net. Meth?ods Mesenchymal stem cells were isolated from human umbilical Wharton’s jelly using tissue adherence method.Their immunophenotypes were detected by flow cytometry . Passage 3 of WJ-MSCs (Wharton’s jelly-mesenchymal stem cells) were inoculated in 4 flasks, denoted as A1, A2, A3, A4 group. TwentyμL of apelin-13 at concentrations of 0, 1×10-6, 10 × 10-6 and 100 ×10-6 mol/L were added to A1, A2, A3 and A4 respectively each day. After being induced for 7 days, cell mor?phology and viability were observed under inverted microscope. Von Willebrand factor (vWF) was examined by immunofluo?rescence and CD31 was identified by flow cytometry. Upon incubating with three dimensional culture medium of hydrogel, those cultured A1, A2, A3 and A4 were renumbered as S1, S2, S3, S4. Again, twentyμL of apelin-13 at concentrations of 0, 1×10-6, 10 ×10-6and 100 ×10-6 mol/L were used to treat S1, S2, S3 and S4 respectively. After 7 days, cell morphology, via?bility and vas-like networks were observed with inverted microscope. Results Our study showed that WJ-MSCs can be in?duced by apelin 13 to differentiate into endothelial cells lineage indicated by positive of vWF staining. Moreover, CD31 expres?sion increases significantly upon apelin-13 addition in a dosage dependent manner. The endothelial cells line formed vas like networks when cultured with three-dimensional medium containing hydrogel. Conclusion This study demonstrated that ape?lin-13 could promote human umbilical cord-MSCs to differentiate into endothelium lineage then to form vascular networks.

BACKGROUND:At present, a lot of research about culture methods for umbilical cord mesenchymal stem cels, but not for the waste of primary system. OBJECTIVE:To explore the best culture method of human umbilical cord mesenchymal stem celsin vitro. METHODS:Human umbilical cord mesenchymal stem cels were prepared by tissue explants method, recorded as initial culture group. The centrifugal fluid and tissue of the primary culture flask were centrifuged and divided into three groups for secondary culture: tissue group, mixed group and pure liquid group. Cel morphology, time for cel acquisition, and yield of primary cels in the four groups were observed; the cel growth curve was analyzed by MTT assay; and cel cycle and phenotype were detected by flow cytometry. RESULTS AND CONCLUSION: The average time for cel acquisition in the initial culture group, tissue group, mixed group and pure liquid group were (15.00±0.45), (7.0±0.3), (8.00±0.25) and (8.00±0.25) days, respectively. The number of cels at first generation was (4.0±0.5)×105, (9.0±0.55)×105, (15.0±0.2)×105 and (7.0±0.33)×105 markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cels can be obtained rapidly and largely through the secondary culture to the primary culture system. T75 culture bottle, respectively. Under the inverted microscope, cels in the four groups were fusiform-like adherent cels, which were in paralel or circinate arrangement. Growth curve, proliferative activity, surface markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cells can be obtained rapidly and largely through the secondary culture to the primary culture system.

Objective To investigate effect and mechanism of Xuebijing administration at various concentrations on ischemia reperfusion injury (IRI) of rabbit limbs.Methods Thirty New Zealand rabbits were divided into control group (n =10),Xuebijing group Ⅰ (n =10) and Xuebijing group Ⅱ(n =10) according to random number table.Rabbit models of IRI in lower extremities were established.Each group received corresponding therapy after reperfusion:rabbits in Xuebijing group Ⅰ were firstly administered 4 ml/kg Xuebijing solution and 6 ml/kg isotonic saline; rabbits in Xuebijing group Ⅱ were administered 2 ml/kg Xuebijing solution and 8 ml/kg isotonic saline; rabbits in control group were simply administered 10 ml/kg isotonic saline.Venous blood samples were collected before reperfusion and at 1 h,2 h,4 h after reperfusion to measure coagulation parameters (APTT,Fib,INR and PT) and biochemical items (ALB,LDH and CK).Results APTT in Xuebijing group Ⅰ presented obvious improvement at 1 h and 4 h after reperfusion as compared with control group (P ＜ 0.01).PT in Xuebijing groups Ⅰ and lⅡ was significantly longer after reperfusion than that before reperfusion (P ＜0.05).Fib level in Xuebijing group Ⅰ was much higher at 4 h after reperfusion than that before reperfusion (P ＜ 0.05).ALB level at 1 hour after reperfusion showed no statistical differences from that before reperfusion in Xuebijing groups Ⅰ and Ⅱ (P ＞ 0.05).LDH and CK levels in Xuebijing group Ⅰ were much lower than those in control group after reperfusion (P ＜ 0.05).Conclusions Xuebijing injection relieves limb IRI,with better effect in Xuebijing group Ⅰ than in Xuebijing group Ⅱ.Therapeutic mechanism may be associated with its involvement in adjusting clotting function and mitigating injury of muscle tissues.

BACKGROUND: Contacting with blood, most of polymer materials lead to different extents of blood coagulation, which limits their clinical application. Therefore, developing polymer materials with excellent anticoagulant property has become a key to clinical study of bioartificial liver materials.OBJECTIVE: To in vitro detect the blood dompatibility of polyacrylamide grafted polypropylene (PP) membrane (PP-g-AAm), a novel artificial liver reactor material.METHODS: Prior to and after modification, hemolytic test, prothrombin time and activated partial thromboplastin time tests of PP membrane were performed; blood platelet CD62P and CD63 expression rates were determined by flow cytometry, and platelet adhesion on PP and PP-g-AAm membranes by scanning electron microscopy.RESULTS AND CONCLUSION: The hemolysis ratio of PP and PP-g-AAm membranes was 1.32% and 1.46%, respectively.Compared with PP-g-AAm membrane, prothrombin time and activated partial thromboplastin time of PP membrane weremarkedly shorter (P < 0.05). CD62P and CD63 expression rates in the PP-g-AAm membrane were significantly lower than PP membrane (P < 0.05). Scanning electron microscopy results revealed that there were obvious changes of platelets adhering to these two membranes, but platelets adhering to PP-g-AAm membrane were fewer than PP membrane. These results indicate that PP-g-AAm membrane exhibits good blood compatibility.

Objective To investigate the optimal condition of lentivirus,which was recombined with marker gene of enhanced green fluorescent protein (Lentivirus-EGFP) transfect human umbilical cord wharton’s jelly-derived mesenchy-mal stem cells (HUWMSCs) and the effect of transfection on the proliferation in HUWMSCs. Methods HUWMSCs were transfected with EGFP by lentivirus vector in vitro via different multiplicity of transfection (MOI) in four different transfec-tion methods (A, B, C and D). The fluorescence expression and the transfection efficiency in different methods were analyzed by both fluorescent microscope and flow cytometry. The proliferation rates of infected HUWMSCs was evaluated by MTT method. Results The transfection efficiency was 10.6%-87.3%after 4 days in all experimental groups, which showed the dose-effect relationship with MOI. Polybrene (5 mg/L) could significantly increase the transfection efficiency (P＜0.05). Re-sults of MTT assay showed that there were significant differences in the proliferation rates of infected HUWMSCs between different transfection methods (P ＜ 0.001). There was better cell proliferation in method A (MOI=10) group and method B (MOI=10) group than that of other groups. Conclusion Method B (MOI=10) is the optimal transfection method in this exper-iment. HUWMSCs could be transfected by lentivirus-EGFP with high efficiency and could stably express transfected gene within 2 weeks, which is a safe and effective gene transfer vector.

Purpose To prepare streptavidin-tagged mouse interleukin-4(mIL-4-SA)bifunctional fusion protein and to study on its bioactivity.Methods The mIL-4 gene was cloned by RT-PCR and cloned into pET21 vector to get mIL-4-SA-pET21 expression plasmid.The mIL-4-SA fusion protein was expressed in BL21 (DE3)host bacteria and purified through the Ni-NTA affinity chromatography and refolded by dilution and dialysis.The effect of mIL-4-SA fusion protein on mouse thymocytes proliferation was evaluated by MTY.Flow cytometric analysis was performed to detect the mIL-4-SA fusion protein on the biotinylated B16F10 tumor cells.Results The mIL-4-SA-pET21 vector was successful by constructed and the mIL-4-SA fusion protein was expressed in BL21(DE3)at about 35%of total bacterial proteins.The purity of mIL-4-SA Was about 95% through Ni-NTA.The mIL-4-SA fusion protein exhibited bifunctional activities,i.e.,stimulative effect for mouse thymocyte proliferation and SA-mediated high-affinity binding to biotinylated cell surfaces(anchoring modified rate Was about 96.69%).Conclusion The mIL-4-SA fusion protein was expected to be developed for the treatment of tumors.

Objective To study and compare the effects and mechanisms between the Xuebijing injection and Danshen injection on ischemic-reperfusion injury of rabbit limb .Methods 30 New Zealand rabbits were randomly divided into control group ,Xuebi-jing group and Danshen group ,with 10 in each group .The Crinnion model of ischemic-reperfusion injury of rabbit limb was used . Each group was treated correspondingly after the blood flow was restored .Xuebijing group and Danshen group were treated with 4 mL/kg Xuebijing injection 2 mL/kg Danshen injection in saline with 10 mL total volume .Control group was treated with 10 mL sa-line .Before releasing clip and 1 h ,2 h and 4 h after reperfusion ,the blood samples were collected for detecting clotting function (APTT ,FIB ,PT-INR ,PT) ,biochemistry items (ALB ,LDH ,CK) ,MDA level and SOD level .Results Compared with control group ,the APTT of Xuebijing group in 1 h and 2 h and Danshen group in 4 h after reperfusion improved significantly (P<0 .05) . The PT of Xuebijing group after reperfusion extended significantly compared with the PT before reperfusion (P<0 .05) .The FIB level of Xuebijing group and Danshen group in 4 h after reperfusion was much higher than before (P<0 .05) .The LDH and CK level of Xuebijing group after reperfusion was much lower than that of control group (P< 0 .05) .The MDA level of Xuebijing group in 2 h and Danshen group in 1 h ,2 h after reperfusion was much lower than that of control group (P<0 .05) .The SOD activ-ity of Xuebijing group in 1-4 h and Danshen group in 2 h after reperfusion was much higher than that of control group (P<0 .05) .Conclusion Xuebijing injection and Danshen injection have the relieving effect on the limb ischemic-reperfusion injury by ad-justing clotting function and decreasing free oxygen radicals .In terms of relieving the injury of muscle tissue ,the effect of Xuebijing injection might be better than Danshen .