Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

Objective To explore the risk factors in children with epilepsy and their effects on attack rate of epilepsy.Methods One hundred and sixty epilepsy patients(patient group,88 boys and 72 girls)and 150 healthy children(control group,72 boys and 78 girls)were selected.All children conformed epilepsy at the west China second hospital were consecutively included in the study for 6 months period.The range of age was from 1 month to 16 years[(7.0?4.7)years old] of patient group children.All children with epilepsy had no-causation seizure for more than twice time and were diagnosed by electroencephalogram.Neurologically normal children in same period,matched for age and sex,visiting the health care clinic were selected as controls.The range of age was from 2 month to 16 years [(6.3?4.5)years old] of control group children.The risk factors examined were febrile convulsions,head trauma,central nervous system infections,abnormal perinatal history,family history of epilepsy and parental consanguinity.The data of patients and controls were obtained from a questionnaire through personal interviews.Details on the patient,family history,and parental age at the time of childbirth were included.Medical records were then reviewed.According to the data type,the statistics were performed with ?2 test and the significance level was the P

OBJECTIVE To discuss the available methods of preventing the mattress pollution in the wards of the(hospital), and to keep the mattresses clean and dry and eliminate the hidden danger of nosocomial infection.(METHODS) The mattress protective covering is used of a new type of textile materials,and then applied into clinic.Selected 100 pieces of mattresses in the wards,divided them equally into two groups at random,the(experimental) group and the control group.After cleaning and pasteurization(surface) sampling and bacterial culturing for every mattress were undertaken.For the experimental group,spread the protective(covering) before the sheet,and for the control group,used the sheet directly.The colony number of each group was compared in the 3rd,7th,and 14th days.RESULTS The mattresses of experimental group were clean,light polluted and with less colonies,and that of the control group were heavy polluted and with more colonies.The comparisons of the total colony number and the number of every sampling point in the 3rd,7th,and 14th days of the two groups showed that there was a(significant) difference(P

Objective To investigate the injuries caused by cholesterol to the vascular endothelial cells (VECs). Methods Different dosage of cholesterol (6.25,12.5,25.0,50.0 mg/L) was used on human umbilical endothelial cell line,ECV304,respectively. LDH activity,nitric oxide and the nitric oxide synthetase activity in the supernatant of cell culture were detected. The concentration of MCP-1 protein in cell culture was detected by ELISA. Results As compared with the normal control cells,a significant increase of LDH activity was found in the cells treated with 50.0 mg/L cholesterol. The NO level decreased in the cells treated by 25.0 or 50.0 mg/L cholesterol. When treated by cholesterol at dose of 6.25,12.5,25.0 or 50.0 mg/L respectively,the NOS activity was greatly decreased and MCP-1 protein was significantly increased in a dose-dependent manner. Conclusion Cholesterol of high concentration could directly injure the structure and partial function of VECs.

Objective To investigate the applicative value of enhanced 3D multi-echo GRE T2*-weighted angiography(ESWAN) sequence phase values in evaluating brain gray nuclei iron content in idiopathic restless legs syndrome(RLS) patients, providing imaging basis in diagnosis and treatment of idiopathic RLS. Methods In our institute from June 2012 to September 2016,forty-five RLS patients were selected as the RLS group, and 45 healthy volunteers as the control group. ESWAN sequence was performed and serum ferritin values were obtained in all patients and volunteers. The raw data of ESWAN was postprocessed , where the phase maps were obtained. Phase analysis was performed on localized brain gray nuclei regions of interest (substantia nigra, red nucleus, dentate nucleus, thalamus, pallidum, putamen and caudate nucleus ) selected on phase maps. Differences between the 2 subject groups were evaluated using ANCOVA including age as a covariate. Results The phase values of the substantia nigra, thalamus, pallidum and putamen in the RLS group were (-0.087 ± 0.021), (-0.053 ± 0.012), (-0.161 ± 0.008), (-0.125 ± 0.019) radians , respectively. The phase values of the substantia nigra, thalamus, pallidum and putamen in the control group were (-0.127 ± 0.007), (-0.066 ± 0.007), (-0.166 ± 0.007), (-0.150 ± 0.010) radians, respectively. There were significant differences between the two groups (F=142.492, 37.988, 10.558, 60.725;P<0.05). Indicating reduced iron content in several regions of brain gray nuclei of the patients with RLS. Serum ferritin concentration between the RLS patients (157.02±95.78)μg/L and healthy controls (175.49 ± 38.65)μg/L was not significant (F=1.353,P>0.05). Conclusions Phase values can make a quantitative assessment of brain gray nuclei iron content in RLS patients, our results supported the hypothesis of reduced brain iron content in RLS patents , which may have an important role in the pathogenesis of the disorder. However, iron content change in some brain regions was not correlated with serum ferritin concentration changes.

Objective To investigate the relationship between plasminogen activator inhibitor-1(PAI-1) activity and PAI-1 promoter 4G/5G polymorphism in patients with type 2 diabetes from the coastal areas of Shandong province.Methods The 4G/5G allele polymorphism in the PAI1 gene promoter region were tested by allele specific PCR in 116 type 2 diabetes and 40 normal controls.The activity of plasma PAI-1 was assayed by chromogenic substrate method.Results The plasma PAI-1 activity in patients was higher than that in controls(P

In this paper, the effect of 5% (V/V) n-alkanes (e.g, n-Heptane, n-Octane, n-Decane, n-Dodecanen-Tetradecane and n-Hexadecane) on the growth and protease production of organic-solvent-tolerant- bacte-rium Bacillus licheniformis YP1 was studied. 5%(V/V) n-alkanes had no effect on the stability of YP1 prote-ase. 5% (V/V) n-alkanes had no notable influence on the yield of strain YP1 but dramatically affected theprotease production. The presence of n-Heptane, n-Octane and n-Decane deeply repressed the protease pro-duction; however n-Dodecane, n-Tetradecane and n-Hexadecane enhanced the protease production promi-nencely. The concentration of n-Tetradecane (1%-8%, V/V) had a direct ration with the protease production.The detailed experiments showed that the notable increase of protease activity appeared at the late logarithmof cultivation compared with the blank. The cell shape of YP1 strain remarkably decreased when grown inthe presence of n-Tetradecane. This is the first report about the effect of n-alkanes on the protease productionby the solvent tolerant bacterium.

Adult ; Female ; Humans ; Logistic Models ; Male ; Mental Health ; statistics & numerical data ; Transients and Migrants ; psychology ; Young Adult

BACKGROUND:The rabbits were widely used as experimental animal models in the research on etiology and pathological mechanism of steroid-induced osteonecrosis of the femoral head. It is stil a valuable and realistic research topic to improve and to innovate the modeling technology nowadays. OBJECTIVE:To improve the modeling technology on osteonecrosis of the femoral head in rabbits induced by glucocorticoid combined with lipopolysaccharide, with the focus on its reduced mortality and the guaranteed successful rate of modeling. METHODS:A total of 28 New Zealand white rabbits were randomly divided into the control group (n=10) and improvement group (n=18). Models of steroid-induced osteonecrosis of the femoral head were established according to different methods. In the improvement group, rabbits were injected with sodium penicilin (5.0 mg/kg) and amikacin sulfate (1.63×104 U/kg) in the left gluteus muscle. Twenty-four hours later, al rabbits were injected with prednisolone acetate (20 mg/kg) in the right gluteus muscle. Forty-eighthours later, 5.0 μg/kg of lipopolysaccharide was intravenously injectedvia the ear. From then on, two injections of prednisolone acetate (20 mg/kg) were respectively performed alternately in the left and right gluteal muscle at an interval of each 24 hours. Sodium penicilin (5.0 mg/kg) and amikacin sulfate (1.63×104 U/kg) were intraperitonealy injected for 2 consecutiveweeks. In the control group, 10 μg/kg lipopolysaccharide was injectedvia the ear vein of rabbit. From then on, prednisolone acetate (20 mg/kg) was intramuscularly injected at an interval of each 24 hours, totaly three times. Benzylpenicilin sodium 20×104 U/rabbit was intramuscularly injected once a week. RESULTS AND CONCLUSION:Rabbit models of steroid-induced osteonecrosis of the femoral head were successfuly established in both groups. Compared with the control group, the mortality was significantly reduced after model establishment in the improvement group, and the bone lacuna and osteonecrosis of the femoral head were apparent. These findings indicated that the improved technology of model establishment of steroid-induced osteonecrosis of the femoral head could be used to aleviate the damage degree on the gluteal muscles, to guarantee the successful rate of modeling, and to noticeably reduce the mortality of rabbits.

Objective: To investigate the therapeutic effects of Tuina (Chinese therapeutic massage) in a knee osteoarthritis (KOA) rat model and its influence on proteins associated with the Wnt/β-catenin signaling pathway. Methods: A total of 32 specific-pathogen-free grade Sprague-Dawley rats were used. Eight rats were randomly selected as the control group (CG). The remaining 24 rats underwent intra-articular injections with 0.2 mL of 4％ papain to prepare the KOA rat models. After the model was established, the 24 rats were randomly and equally assigned to 3 groups, including a model group (MG), a Tuina group (TG), and a positive medicine group (PMG), with 8 rats in each group. The Lequesne score was applied to evaluate the success of model development. After the model was successfully established, the CG did not receive any intervention, and the TG was treated with local, clockwise annular Rou-Kneading around the knee joint with the thumbs. The pressure in the longitudinal direction was 3 N, and the frequency was designed to be 120-140 times/min for 15 min, followed by flexing the joint 10 times. The PMG was intragastrically administered with celecoxib [24 mg/(kg·bw)] every day. These interventions were performed once a day, 6 d per week, for a total of 4 weeks. After treatment, the Lequesne score was applied again to assess the severity of the KOA in the rats; hematoxylin-eosin (HE) staining and a mixture of equal volumes of aqueous solutions of safranin O-fast green were used to stain and observe the cartilage morphology and structure; the modified Mankin score was applied to evaluate the pathology; enzyme-linked immunosorbent assay method was used to quantify the C-telopeptide fragments of type Ⅱ collagen (CTX-Ⅱ) and cartilage oligomeric matrix protein (COMP); Western blotting was then applied to quantify Wnt4, β-catenin, matrix metalloproteinase 13 (MMP-13), and bone morphogenetic protein 2 (BMP-2) protein expression; immunohistochemistry was conducted to determine the percentage of collagen type X (ColX)-positive cells. Results: The Lequesne score of the TG and PMG was both lower than that of the MG (P<0.01); the HE staining, safranin O-fast green stained morphology and structure, and modified Mankin scores of the TG and the PMG were also better than those in the MG (P<0.01). Compared with the CG, the amounts of CTX-Ⅱ and COMP in the serum were significantly increased (P<0.01); the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins in the cartilage tissue was significantly increased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly increased (P<0.01) in the MG. In comparison with those in the MG, the amounts of CTX-Ⅱ and COMP were significantly decreased (P<0.01), the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins was significantly decreased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG and PMG. Compared with the PMG, the contents of CTX-Ⅱ and COMP and the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins were decreased (P<0.05 or P<0.01); the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG. Conclusion: Tuina can relieve the degeneration of KOA, and the mechanism may be related to the down-regulation of the Wnt/β-catenin signaling pathway, the decrease in MMP-13 and BMP-2 protein expression, the reduction in chondrocyte extracellular matrix degradation, and slowing down the terminal cell differentiation.