Differentiation of Mesenchymal Stem Cells Into Dopaminergic Neuron-like Cells in vitro

To explore the way to induce mesenchymal stem cells (MSCs) to differentiate into dopaminergic neurons in vitro. Methods MSCs were obtained from rat bone marrow, cultured and passaged. MSCs used in this experiment had multipotency, which was indirectly proved by being induced to differentiate into chondrocytes and adipocytes. MSCs were cultured in medium containing 0.5 mmol/L IBMX for 2 days. Then the medium was replaced with induction medium, which contained GDNF, IL-1β, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments. The surface markers of the differentiated neurons, such as NSE, nestin, MAP-2a, b and TH were detected by immunocytochemistry and Western blot after MSCs were cultured in induction medium for 7 days and 15 days. Results MSCs differentiated into neural progenitors and expressed nestin after MSCs were incubated with medium containing IBMX for 2 d. After the medium was replaced with induction medium containing many inducing agents, MSCs differentiated into neuron-like cells and dopaminergic neuron-like cells and expressed NSE, MAP-2a, b and TH. The percentage of NSE-positive cells, MAP-2a, b-positive cells and TH-positive cells was 30.032±2.489%, 41.580±5.101% and 34.958±5.534%, respectively after MSCs were induced in medium containing GDNF, IL-1β, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments for 15 days. Conclusion MSCs can differentiate into dopaminergic neuron-like cells and are a new cell source for the treatment of neurodegeneration diseases and have a great potential for wide application

Enhanced antitumor efficacy on hepatoma-bearing rats with adriamycin-loaded nanoparticles administered into hepatic artery

AIM: To investigate the antitumor activity of adriamycin (ADR) encapsulated in nanoparticles (NADR) and injected into the hepatic artery of hepatoma-bearing rats.METHODS: NADR was prepared by the interfacial polymerization method. Walker-256 carcinosarcomas were surgically implanted into the left liver lobes of 60 male Wistar rats, which were divided into 4 groups at random (15 rats per group). On the 7th day after implantation, normal saline (NS), free ADR (FADR), NADR, or ADR mixed with unloaded nanoparticles (ADR+NP) was respectively injected via the hepatic artery (i.a.) of rats in different groups. The dose of ADR in each formulation was 2.0 mg/kg body weight and the concentration was 1.0 mg/mL. Survival time, tumor enlargement ratio, and tumor necrosis degree were compared between each group.RESULTS: Compared with the rats that received NS i.a.,the rats that received FADR or ADR+NP acquired apparent inhibition on tumor growth, as well as prolonged their life span. Further significant anticancer efficacy was observed in rats that received i.a. administration of NADR. Statistics indicated that NADR brought on a more significant tumor inhibition and more extensive tumor necrosis, as compared to FADR or ADR+NP. The mean tumor enlargement ratio on the 7th day after NADR i.a. was 1.106. The mean tumorbearing survival time was 39.50 days. Prolonged life span ratio was 109.22% as compared with rats that accepted NS.CONCLUSION: Therapeutic effect of ADR on liver malignancy can be significantly enhanced by its nanopaticle formulation and administration via hepatic artery.

Relationship between bulbocavernosus reflex, patellar reflex and rehabilitation during acute stage of cervical spinal injury

AIM:To investigate the relationship between bulbocavernous reflex(BTR),patellar reflex(PTR) and rehabilitation during acute stage of cervical spinal injury.METHODS: 73 cases of a class frankel cervical spinal injury patients were examined with BTR and PTR and recorded the time of the two reflexes,then follow up after 6 months to monitor recovery situation. RESULTS:There were 28 cases of positive BCR within the first 72 h (38%),7 cases of improved conditions(25%); there were 13 cases of PTR positive (17%),13 cases of improved status(100%). CONCLUSION:BCR and PTR are the sign of improved status and better rehabilitation during the acute stage.

Signal transduction of gap junctional genes,connexin32,connexin43 in human hepatocarcinogenesis

AIM: To investigate gap junctional intercellular communication (GJIC) in hepatocellular carcinoma cell lines, and signal transduction mechanism of gap junction genes connexin32(cx32),connexin43(cx43) in human hepatocarcinogenesis.METHODS: Scarped loading and dye transfer (SLDT) was employed with Lucifer Yellow (LY) to detect GJIC function in hepatocellular carcinoma cell lines HHCC, SMMC-7721and normal control liver cell line QZG. After Fluo-3AM loading, laser scanning confocal microscope (LSCM) was used to measure concentrations of intracellular calcium [Ca2+]i in the cells. The phosphorylation on tyrosine of connexin proteins was examined by immunoblot.RESULTS: SLDT showed that ability of GJIC function was higher in QZG cell than that in HHCC and SMMC-7721 cell lines. By laser scanning confocal microscopy, concentrations of intracellular free calcium [Ca2+]i was much higher in QZG cell line (108.37 nmol/L) than those in HHCC (35.13nmol/L) and SMMC-7721 (47.08 nmol/L) cells. Western blot suggested that only QZG cells had unphosphorylated tyrosine in Cx32 protein of 32 ku and Cx43 protein of 43ku; SMMC-7721 cells showed phosphorylated tyrosine Cx43 protein.CONCLUSION: The results indicated that carcinogenesis and development of human hepatocellular carcinoma related with the abnormal expression of cx genes and disorder of its signal transduction pathway, such as decrease of [Ca2+]i,post-translation phosphorylation on tyrosine of Cx proteins which led to a dramatic disruption of GJIC.

Effect of AXIS lateral mass screw-plate internal fixation system on functional recovery of spinal cord in fracture of lower cervical vertebrae

AIM:To evaluate the methods and results of the AXIS lateral mass screw plate system in the treating of lower cervical spine injury.METHODS:29 cases of lower cervical vertebrae injury were fixed with AXIS system.All of them were followed up for more than 1 year.RESULTS:All cases had a good bone union without malformation.We found no complication of the injury to the vertebral arteries or nerves.There was no loosening of the plate and screw. CONCLUSION:AXIS lateral mass screw plate system has the characteristic of stable,simple and safe and is suitable for the treatment of lower cervical spine injury.

Capacity of bone induction of compound material of decalcified bone matrix combined with rhBMP-2 and impregnated with bone cement

AIM:To analyze bone inductive capacity of the compound material of decalcified bone matrix combined with rhBMP 2 and impregnated with bone cement.METHODS:To assess the experimental study, histological and Masson’s methods were used.RESULTS:The effects of compound material on the induction of bone formation were investigated in NIH mouse models.It was observed that in the with rhBMP 2 group, mesenchymal cells gathered in the implanted material at the 7th day postoperation,chondrogenesis were found at 14 to 21 days after implantation,new bone formation were observed at about 21 to 28 days after surgery and the DBM particles were absorbed by the new generated tissues gradually.CONCLUSIONS:The compound material of DBM combined with rhBMP 2 and impregnated with bone cement could induce the proliferation and migration of mesenchymal tissues that could be differentiated into cartilage and formed new bone finally.The new bone could absorb DBM particles gradually.The compound material had fair capacity of bone induction.

Effects of 24h ultra-marathon on biochemical and hematological parameters

AIM: To analyze detailed changes in hematology and biochemistry tests parameters before and after a longdistance race in ultramarathon runners.METHODS: Blood samples of 11 participants were obtained for standard analysis before, immediately after, two days after and nine days after the 2002 International Ultramarathon 24 h Race and the International Association of Ultrarunners (IAU) Asia 24 h Championship.RESULTS: Total bilirubin (BIL-T), direct bilirubin (BIL-D),alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) increased statistically significantly(P<0.05) the race. Significant declines (P<0.05) in red blood cell (RBC), hemoglobin (Hb) and hematocrit (Hct)were detected two days and nine days d after the race.2 d after the race, total protein (TP), concentration of albumin and globulin decreased significantly. While BIL, BIL-D and ALP recovered to their original levels. High-density lipoprotein cholesterol (HDL-C) remained unchanged immediately after the race, but it was significantly decreased on the second and ninth days after the race.CONCLUSION: Ultra-marathon running is associated with a wide range of significant changes in hematological parameters, several of which are injury related. To provide appropriate health care and intervention, the man who receives athletes on high frequent training program high intensity training programs must monitor their liver and gallbladder function.

Effects of exercise on lipid metabolism and musculoskeletal fitness in female athletes

AIM: This study investigated the effects of intense training on lipid metabolism, bone metabolism and bone mineral density (BMD) in female athletes. METHODS: Sixty-six female subjects participated in this study,age ranging from 18 to 55 years. The sample group included thirty-six athletic subjects and the control group comprised thirty non-athletic individuals. Five athletes competed with national level (5/36) and nine non-athletic subjects (9/30) were postmenopausal women. The assessment items induded body composition, radius BND, calcaneus BND, lung function,muscular endurance, renal and liver function, bone marker assay and hormone status. All data were analysed, using SPSS 10.0 software, and were presented as mean rank statistical difference, using bhe Kurskal-Wallis (K-W) test. Alter that the non-parameter statistics were used. Either Kvalue or Pvalue below 0.05 was considered significant.RESULTS: Urine deoxypyridinoline/creatinine (Dpd/Cre) levels increased significantly (5.93±2.31 vs 6.85±1.43,/K<0.01),sit-reach (29.30±9.48 cm vs 41.31±9.43 cm, K<0.001,P<0.001), 1 minute sit-ups with bended knees (1 min sit-ups) (17.60±9.34 count vs30.00±10.38 count,K<0.001,P<0.001), and vertical jump (25.27±6.63 cm vs 34.69±7.99cm,K<0.001, P<0.001) improved significantly in the athletes group. The athletes group also had a significantly increased level of estriol (E3) (0.14±0.13 pg/mL vs 0.07±0.04 pg/mL,K<0.01, P<0.01), radius BMD (1.37±0.49 gm/cm^2 vs 1.19±0.40gm/cm^2,K<0.05) and calcaneus BMD (0.57±0.17 gm/cm^2 vs-0.20±0.17 gm/cm^2, K<0.01, P<0.05) compared with those of the controls. The high density lipoprotein (HDL)(65.00±14.02 mg/dL vs 52.26±4.84 mg/dL,K<0.05, P<0.05)was significantly lower in postmenopausal inactive athletes(5/36) than premenopausal active athletes (31/36). On the other hand, low-density lipoprotein (LDL) (98.35±23.84 rng/dL vs 131.00±21.63 mg/dL, K<0.05, P<0.01), cholesterol (CHO)(164.03±27.01 mg/dL vs 193.00±23.48 mg/dL, K<0.05,P<0.05), triglyceride (TG) (63.00±26.39 mg/dL vs 147.00±87.21 mg/dL, K<0.01), body fat % (BF%) (28.16±4.90% vs 34.84±4.44%, K<0.05, P<0.001) and body mass index (BMI)(21.98±2.98 kg/m^2 vs 26.42±5.01 kg/m^2, K<0.05, P<0.001)were significantly higher in postmenopausal inactive athletes(5/36) than premenopausal active athletes (31/36). TG(90.22±39.82 mg/dL vs 147.00±87.21 mg/dL), CHO(186.44±124.90 mg/dL vs 193.00±123.48 mg/dL) were higher,but the HDL was significantly lower (62.18±110.68 mg/dL vs 52.26±14.84 mg/dL, P<0.05) in postmenopausal athletes(5/36) group than in postmenopausal control group (9/30). CONCLUSION: Postmenopausal athletes (5/36) who nolonger took competing exercises had reduced levels ofphysical activity, faced increased risk of cardiovascular disease compared to active athletes (31/36) and the postmenopausal controls (9/30). We may thus concluded that long term exercise effectively improves musculoskeletal fitness and prevents BMD loss in female athletes.

Effects of Extracellular ATP and Its Receptors on Peripheral Nerve Regeneration in SD Rats

To explore the effect of the extracellular ATP and its receptors on axon regeneration in the sciatic nerve defect in rats, 0.5 cm defect of the sciatic nerve was made and repaired with long arm silicon tube like a 'Y' type. The single arm of the silicon tube was sutured to proximal of the sciatic nerve. 10 μl 1 mmol/L ATP in physiological saline was injected into the left chamber of the silicon tube (experimental group) and physiological saline injected into another silicon tube as a control group. In another model 1 mmol/L 10 μl ATP and 1 mmol/L 10 μl ATP+0.2 mg/ml suramin were injected respectively into two arms of the silicon tube. After 4 and 8 weeks the specimens were obtained from the silicon tube for examining the axon regeneration histologically and image analysis. All the regeneration axons grew into the silicon tube containing the ATP, but there was no axon regeneration in the silicon tube containing the ATP+Suramin or physiological saline. It was demonstrated that extracellular ATP had a powerful attraction to the regenerated axon of peripheral nerve. The suramin inhibited the axon induction of the extracellular ATP completely via the ATP receptors.

EFFECTS OF TGF β1 AUTOCRINE BLOCKAGE ONOSTEOSARCOMA CELLS

Objective To evaluate the effects of transforming growth factor β1 (TGF β1) autocrine blockage on proliferation activity and drug sensitivity of osteosarcoma. Methods Northern blot, MTT determination, and 3H thymidine incorporation were used to investigate the effects of antisense TGF β1 gene on osteosarcoma. Results The proliferation of osteosarcoma cells transfected by antisense TGF β1 gene was suppressed markedly, and adriamycin sensitivity was significantly increased. Conclusion Blockage of osteosarcoma cells TGF β1 autocrine loop inhibits cell proliferation and enhances chemother-apy sensitivity.

ANALYSIS AND RESEARCH OF COMPUTER-AIDED MODEL OF HIP JOINT BASED ON REVERSE ENGINEERING

Former research work about the modeling of hip joint focus on the uppersegment of femoral, and assumes the acetabulum cup is sphere concave, and the acetabulum prosthesesis semisphere. A method of acquiring the point data on the surface of the hipbone using the reverseengineering technology is presented. After analyzing the acetabulum surface fitting error, arotation ellipsoid CAD model is applied to fit the acetabulum surface, and then optimizationtechnique is used to find the geometric parameters of the model. The fitting error between thesphere and rotation ellipsoid is compared and gets the result that the fitting error of rotationellipsoid is smaller than sphere, and the rotation ellipsoid can describe the shape of theacetabulum better.