BACKGROUND:Different structures of matrix models, such as grating, holes and pil ars make different effects on the differentiation of neural stem cel s. OBJECTIVE:To explore the effects of the diameter and spacing, known as physical signals of micro pil ars on neural stem cel differentiation. METHODS:Micro pil ars with different diameters and spacing, both of which had four dimensions of 2.5, 5, 10 and 20μm, were fabricated on silicon substrates by photolithographic method. Purified primary neural stem cel s were incubated on the each micro pil ar for 7 days in vitro. Then the differentiation of neural stem cel s into neuron-like cel s was observed using immunofluorescence staining and quantitative real-time PCR. RESULTS AND CONCLUSION:When the diameters of the micro pil ars were constant and the spacing of micro pil ars varied in the range of 2.5-10μm, the differentiation rate of neural stem cel s increased with the spacing increase. When the spacing was invariable and the diameters changed in the range of 2.5-20μm, the differentiation rate of neural stem cel s declined with the diameter increase. Especial y, the micro pil ars with 2.5μm diameter and 10μm spacing significantly promoted the differentiation of neural stem cel s into neuron-like cel s. These results show that specific micro pil ars with smal diameters and large spacing facilitate the differentiation of neural stem cel s, thus providing guidance for developing tissue-engineered scaffolds.

Objective To evaluate safety and clinical significance of single balloon enteroscopy (SBE) for small intestinal diseases. Methods Data of 83 patients with suspected or known small intestinal diseases, who underwent SBE from March 2009 to July 2010, were reviewed in terms of preparation time,procedure time, detection rate and complication occurrence. Results The 83 patients included 37 cases of digestive tract bleeding, 38 chronic abdominal pain, 1 chronic diarrhea, 2 fever and 5 incomplete ileus. A total of 94 procedures of SBE were performed, including oral route in 46 patients, anal route in 26 and both routes in 11. Excluding 6 cases with endoscopic therapy, the mean procedure time of oral approach was 29.6 ± 10. 3 min, and that of anal route was 57.1 ± 15.6 min. Abnormalities were detected in 57 ( 68.7% )of the 83 patients, with detection rate of 81.1% (30/37) in digestive tract bleeding with unknown reason,57. 8% (22/38) in chronic abdominal pain of unknown reason, 50. 0% (1/2) in fever of unknown reason and 80. 0% (4/5) in incomplete ileus. Peutz-Jeghers syndrome was diagnosed in 6 patients and endoscopic polypectomy was performed, with complicated bleeding in one patient. No other procedure-related complications were observed. Conclusion SBE is well-tolerated and safe for diagnosis of small intestine diseases,with easy manipulatiou, short procedure time, high detection rate and satisfactory location of intestinal hemorrhagic lesions.

Objective To evaluate the security of big animals with a new bioartificial liver system .Methods Six Tibet pigs re-spectively received treatment of hybrid artificial liver and simple bioartificial liver ,observed and recorded the vital signs ,venous pressure ,transmembrane pressure and slurry pot pressure each hour ,and collected blood to make endotoxin and bacterial culture test in the zero hour ,fourth and eighth hour .Results Compared with the zero hour ,venous pressure ,transmembrane pressure ,slur-ry pot pressure were much higher in the fifth hour (P< 0 .05) ,and there were no significant difference in the rest of other time points(P>0 .05) .The mean arterial pressure and respiratory rate in all time point showed no significant changes (P>0 .05) .Com-pared with the zero hour ,the heart rate was much lower in the second hour (P<0 .01) .The values of blood endotoxin were less than 0 .5 EU/mL in the zero hour ,fourth and eighth hour from beginning ,and the bacterial culture test showed no growth of bacteria . Conclusion The experiment of big animals with a new bioartificial liver system was safe ,the efficacy in the treatment of hepatic failure could be assessed further .

Objective To study the radiosensitizing effect of 2'-hydroxyflavanone (2'-HF) on prostate cancer cells,and to preliminarily investigate its mechanism.Methods Colony formation assay,tert-butylhydroperoxide (TBHP) oxidative stress assay,Hoechst staining,and apoptosis flow cytometry using Annexin V-FITC and propidium iodide (PI) were performed to measure the impact of 2'-HF on the radiosensitivity of VCaP prostate cancer cells.Western blot was used to determine the effects of 2'-HF on expression of AKT,phosphorylated AKT (p-AKT),and aldo-keto reductase 1 C3 (AKR1 C3) in VCaP cells and preliminarily investigate the mechanism.Data were analyzed by t test and factorial analysis of variance.Results The results of colony formation assay indicated that after exposure to radiation,VCaP cells treated with 2'-HF had a significantly lower proliferation level than cells in the control group (P=0.010),yielding a sensitization enhancement ratio of 1.19.The resuhs of TBHP oxidative stress assay suggested that VCaP cells treated with 2'-HF had significantly weaker anti-oxidative capacity than cells in the control group (P=0.015).Hoechst staining and apoptosis flow cytometry with Annexin V-FITC and PI indicated that 2'-HF treatment plus irradiation significantly enhanced apoptosis in VCaP cells (P=0.001.The results of Western blot suggested that 2'-HF treatment significantly inhibited the protein expression of p-AKT and AKR1C3 in VCaP cells (P=0.013 and P=0.016).Conclusions 2'-HF can enhance the radiosensitivity of prostate cancer cells,which is probably associated with its inhibitory effects on AKT pathway and AKR1C3 expression in prostate cancer cells.

Objective To evaluate the novel hybrid liver and renal support system the safety and efficacy of the treatment of Tibet mini pig model of renal failure .Methods Five Tibet miniature pig bilateral renal artery ligation approached to construct the models of renal failure ,treatment group was treated with therapy (CRRT mode) of the novel hybrid liver and renal support system after modeling ,while the control group received no treatment .Drawning venous blood endotoxin ,biochemical tests and aerobic anae‐robic culture when in the treatment of 0 ,4 ,8 h;then we observed the changes of biochemical indicators in Tibet mini pigs before and after treatment ;we recorded machine pressure monitoring indicators every 2 hour during treatment .Results The model of renal failure of two experimental animals were successfully constructed .The general condition of treatment group was improved after treated by the novel hybrid liver and renal support system ,while the control groups continued to deteriorate .The experimental ani‐mals did not appear abnormal ECG during the experiment .Two groups of animals showed no statistics significant difference (P>0 .05) in the period of 0 h .In the remaining period ,heart rate and respiratory rate in treatment group was significantly reduced(P<0 .05) ,while oxygen saturation and mean arterial pressure were significantly increased compared to control group(P<0 .05) .There was no differences of biochemical indicators between two groups before modeling (P>0 .05);after modeling both groups biochemi‐cal indicators was significantly increased compared with baseline (P<0 .05);after treatment of the hybrid liver and renal support systems in the treatment group ,the biochemical parameters were significantly decreased compared with baseline ,the difference was statistically significant different(P<0 .05) ,while the control group did not change significantly .Blood endotoxin measured results were less than 0 .5 EU/mL ,arteriovenous end aerobic anaerobic culture showed no bacterial growth .Pathological examination showed significant swelling of the renal tubular epithelium and tubular necrosis .Conclusion The novel hybrid liver and renal sup‐port system for the treatment of Tibet mini‐pig model of renal failure is safe and effective ,and it can be further attempts for clinical trials .

Objective Introduce a technique for creating 3-dimensional (3D) brain models of subdural electrodes , ideal for demonstrating the space-relationship of seizure localization and eloquent cortices , and discuss its usefulness for presurgical evaluation of epileptic patients. Methods Patients with medically intractable epilepsy were underwent a thorough preoperative MR brain scan including a T1-weighted high-resolution 3D sequence. Intraoperative photographs were taken with the digital camera. After the surgical implantation of subdural electrodes for epileptic zone localization , the thin-slice CT scan of the electrodes were taken and coregistered to preoperative brain 3D MR images in the Medtronic StealthMerge software environment. After the iEEG monitoring , multiple habitual seizures were recorded , eloquent areas were also identified by electrical stimulation of the cortex. The epileptic zone and the eloquent areas were marked on the subdural electrodes respectively. Then, 3D tessellations of the epileptic zone and the eloquent areas were rendered. Results Six patients (4 male, 2 female) were enrolled in this study. The mean time of iEEG monitoring was 13.7 days. After the coregistration of postoperative CT with preoperative 3D MRI , the 3D stereoscopic reconstruction provided an accurate representation of the implanted electrodes with highly detailed visualization of the underlying anatomy. The visual comparison between 3D reconstructions and intraoperative photographs indicated a good correspondence. The patients were followed for 3 to 6 months after the secondary operation , and had continuing improvement in seizure control. Conclusions The results indicate that the 3D reconstruction of subdural electrodes can reveal the precise localization of subdural electrodes , and can be useful for the presurgical evaluation of epileptic patients.

Objective To observe the therapeutic effect of glycosaminoglycans(GAGs)on diabetes mellitus with early nephropathy in order to find a new therapeutic approach to diabetic nephropathy. Methods 60 cases of type 2 diabetic nephropathy(albuminuria:30 to 300mg/24h,male/female:38/22,ages:43-70 years old, course of disease:1-30 years)without hypertension were selected. Some indexes were analyzed before and after administration of regular therapy in routine group or glycosaminoglycans group. The elderly group and non elderly group of diabetes nephropathy were compared. When the metabolism is stable, the levels of endothelin (ET), Netricoxide (NO)and urinary albumin excretion rate(UAER)were measured. Results After three months treatment, the levels of UAER were decreased significantly in both GAGs group and routine group(P＜0.01).After three months, UAER was decreased step by step, and there was no difference between the two groups. The levels of UAER had no change in regular group and there was significant difference between this group and the other two groups. In GAGs group, the levels of whole blood viscosity of medium shear rate 1,whole blood viscosity of medium sheer rate 2,whole blood viscosity of low shear rate were declined and serum NO increased significantly; that of plasma viscosity, whole blood reduced viscosity, ET were all decreased to some degree. Conclusion GAGs has the therapeutic effect on type 2 diabetic nephropathy patients with microalbuminuria because of decreasing UAER and reversing the development of DN. The benefit was positively correlated with the time of taking glycosaminoglycans.

Objective:Preparation of monoclonal antibodies against Carprofen,which lays a foundation for the rapid detection of Carprofen.Methods:Via an active ester method,Carprofen was conjugated to bovine serum albumin (BSA) as immune antigen and ovalbumin (OVA) as detection antigen,respectively.Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the mice immunized with Carprofen-BSA.Hybridomas 51C3 secreting antibodies against Carprofen were obtained and subcloned.Results: Ascites of monoclonal antibodies (McAb) were prepared by injecting cells of hybridoma 51C3 into mice abdomen.The titer of purified McAb was 3.2×105and the McAb was IgG1 subtype.McAb was highly specific for Carprofen and the cross-reactivity (CR50%) was less than 0.04% with Ibuprofen, Ketoprofen, Naproxen, Feinuoluofen, Indoprofen, Fenbufen respectively.The sensitivity of the McAb to carprofen was 0.32 ng/ml and the IC50value was 0.882 ng/ml.The recoveries of Carprofen in the pork samples were from 81.4% to 104.4 % and coefficients of variation were from 16.3% to 25.8%.Conclusion:All results indicate that the McAb 51C3 is suitable to develop an immunoassay colloidal gold rapid dipstick test.

Objective To investigate the effect of intravenous fluid infusion combined with water throuth gastrointestinal tract in treating the patients with hyperosmolar hyperglycemic state(HHS).Methods 30 HHS pa- tients were recruited.All the patients were given water throuth gastrointestinal infusion while they were administrat- ed continuously intravenous infusion.Laboratory parameters such as serum natrium,serum potasium,serum glucose and serum osmolarity ect were monitored at the admission and after treatment.Results Serum glucose and serum osmolarity of HHS patients were decreased smoothly at the speed of less than 3 mmol?L~(-1).h~(-1)during the first 12 hours after treatment.After 48 hour-treatment,serum natrium,serum potassium and serum osmolarity recovered to normal levels except 2 deaths,serum glucose decreased to(10.8?5.2)mmol/L.Conclusion Intravenous fluid infu- sion combined with water throuth gastrointestinal tract for the patients could lower smoothly serum glucose and serum osmolarity and decrease the mortality of the HHS patients.

Objective To identify the effect of six components of salviae miltiorrhizae on the metabolism of fluvastatin in the human liver microsomes (HLMs) and to further explore the effect of cryptotanshinone and dihydrotanshinone Ⅰ on the metabolism of fluvastatin based on the polymorphism (* 1,* 2,* 3) of cytochrome P450 2C9 (CYP2C9).Methods Establishing the incubation system of HLMs and recombinase and determining the concentration of fluvastatin by HPLC.To calculate the enzyme kinetic parameters,half maximal inhibitory concentration (IC50) and inhibition constant (Ki) so as to compare the impact of different CYP2C9 mutation site on the metabolism of fluvastatin and the influence of components of salviae miltiorrhizae on the metabolism process.Results It was shown that cryptotanshinone and dihydrotanshinone Ⅰ exerted significant inhibition on the metabolism of fluvastatin and the residual activity were 30.24％,14.52％ respectively when the concentrations were 10 μmol · L-1 in the HLMs.However,other exhibited no inhibitory effect.The michaelis constant (Km),maximum velocity (Vmax) and intrinsic clearance (CLint) of CYP2C9 * 1,* 2,* 3 recombinase-catalyzed fluvastatin was (1.56±0.21),(1.79±0.25),(2.32±0.43)μmol· L-1,(41.75±1.67),(26.55±1.08),(12.85±0.79) pmol · min-1 · mg-1 and 26.71,14.86,5.53 μL · min-1 · mg-1 respectively.It was indicated that the metabolic activity of CYP2C9 * 1 was stronger than * 2,* 3.The IC50 value of CYP2C9 * 1,* 2,* 3 recombinase-catalyzed fluvastatin by cryptotanshinone and dihydrotanshinone Ⅰ was (1.74 ± 0.41),(2.64 ± 0.81),(3.17 ± 0.32) μmol · L-1 and (0.19 ±0.09),(0.56 ±0.16),(1.52 ±0.34) μmol · L-1.The IC50 and Ki value of CYP2C9 * 1 was less than * 2 and * 3 but there was statistical significance between the * 1 and * 3 with respect to the difference of IC50 and Ki.Conclusion The mutation site of 430(C ＞ T) and 1075 (A ＞ C) could influence the metabolic activity of CYP2C9.The cryptotanshinone and dihydrotanshinone Ⅰ exerted strong inhibitory effect against CYP2C9 recombinase-catalyzed fluvastatin and their inhibitions on CYP2C9 * 1,* 2,* 3 recombinase-catalyzed fluvastatin were different.