AIM: To investigate the synergistic effect of resveratrol and 5-fluorouracil on osteosarcoma CD133+ cell subset.METHODS: Human osteosarcoma cell line MG-63 CD133+ cell subset and the corresponding CD133-cell subset were treated with resveratrol and 5-fluorouracil.After treatment, the viability of MG-63 cells was measured by MTT assay.The apoptosis of MG-63 cells was analyzed by flow cytometry.Activation of caspase-9 and caspase-3, the expression of Apaf-1, and the release of cytochrome C were evaluated by Western blot.The interaction between Apaf-1 and pro-caspase-9 was detected by co-immunoprecipitation.RESULTS: The cell death and apoptosis of MG-63 CD133+ cell subset induced by 5-fluorouracil were significantly weaker than those in the corresponding MG-63 CD133-cell subset.However, co-treatment with resveratrol significantly enhanced the effect of 5-fluorouracil on inhibiting the viability of MG-63 CD133+ cell subset.Mechanically, treatment with resveratrol upregulated the expression of Apaf-1.Transfection with Apaf-1 siRNA abolished the synergistic effect of resveratrol and 5-fluorouracil in MG-63 CD133+ cell subset.In addition, the results of co-immunoprecipitation indicated that the combination of resveratrol and 5-fluorouracil significantly induced the formation of Apaf-1/pro-caspase-9 complex, leading to the activation of caspase-9 in MG-63 CD133+ cell subset.CONCLUSION: Resveratrol enhances 5-fluorouracil-induced apoptosis of osteosarcoma CD133+ cell subset by promoting the formation of Apaf-1/caspase-9 complex.

Objective To solve the encountered problem of lack of space and dyskinesia of detector in dual energy X-ray absorptiometry system, Methods The method used PartitionMagic8.0 software to partition space in hard disk and added the load to keep the detector balance for moving, Results The original software can run in hard disk, and the pulleys are not gotten force from side. Conclusion The reason of problem is analyzed to find the method that can reduce the percentage for system failure and improve precision and accuracy of data.

The objective of this paper is to find out a way that calculate the consumed peak value of oxygen in medical central oxygen supply system,which is the basis for discussion of purchasing hospital oxygen generating system.The author established a math model by poisson distribution to describe the consumed peak value of oxygen in medical central oxygen supply system to find out a convenience way to calculate the consumed peak value of oxygen.

Objective To determine the indexes for evaluating medical central compressing air sypply system.Methods According to the gas equation,pressure calculation was perfomed with the pressure variation as the monitoring subject.Results Such parameters could be used for monitoring medical central compressing air supply system as as average velocity & instanteous velocity of gas consumption,air compressor's average exhaust velocity & exhaust velocity at start-stop point,and pressure-exhaust velocity curve of air compressor.Conclusion The method can be applied to enhancement of quality management & maintenance of the equipment.

Objective The tissue-engineered composite graft was formed with induced marrow-derived stromal cells (MSCs)and PLGA double-layer scaffold. The effectiveness of this graft for the repair of osteochondral defects in the knee of rabbits was investigated. Methods MSCs were isolated from 20 adult rabbits with density gradient centrifugation and was divided into two groups. In group A, the MSCs were cultivated with regular medium. In group B they were cultivated with chondrogenic differentiation medium. The mRNA of MSCs and articular cartilage cells were extracted, and the expression of mRNA for type Ⅰ and Ⅱ collagen was tested by RT-PCR. The distribution and compound of MSCs with PLGA double-layer scaffold was examined with scanning electron microscopy. 28 adult rabbits were divided into 3 groups, osteochondral defect of 3.5 mm in diameter and 3 to 4 mm in depth were created in the patellar groove. Group A (10 rabbits), the MSCs cultivated with regular medium was grafted into the defects. In group B (10 rabbits), the MSCs cultivated with chondrogenic differentiation medium was grafted into the defects. In group C (8 rabbits), the defects were repaired with autologous osteochondral grafts as control. Specimens were harvested at 4th, 8th, 16th and 24th week post operation respectively, histological examination was performed and graded. Results For the MSCs cultivated with regular medium, the expression of mRNA for type Ⅰ collagen was found with RT-PCR, but no expression for Ⅱ collagen was found. For the induced MSCs, the expression of mRNA both for type Ⅰ and type Ⅱ collagen were found. The adhesion and growth of MSCs on the PLGA double-layer scaffold were well visualized with scanning electron microscopy, and some cells were found in the deep porotic area. For the specimens of group B, no significant difference was found comparing with normal cartilage at 24th week, and the specimens were defined as matured hyaline-like cartilage(4/6)with histological examination, superior to those specimens of group A (1/4). Conclusion The MSCs have osteogenic and chondrogenic potentiality. Combined with PLGA double-layer scaffold, it can be served as seeded cell to form tissue-engineered composite grafts, which can be used to repair osteochondral defects in rabbit models.

Objective To develop a method to prepare human articular cartilage derived microcarrier for both rapid propagating chondrocytes and being used as scaffold to support chondrogenesis. Methods Human articular cartilage was crushed into small pieces by muller after lyophilization, and sorted through two different meshes to collect only those specimens measuring 150-200 microns. Then, in turn, the specimens were subjected to 0.25% trypsin at 37 ℃ for 24 hours and 1% Triton X-100 for 72 hours, respectively. The specimens were observed by inverted phase contrast microscopy, and assessed by staining with haematoxylin-eosin, safranin-O (for GAG), as well as by the immunohistochemistry of aggrecan, collagen type Ⅱ. The microcarriers were seeded with human chondrocytes after being irradiated by 60Co. Results Using inverted phasecontrast microscope, the freezing-dry cartilage particles were observed as yellow, different shapes, and their surfaces were uneven, and with many pits. After treating with trypsin and Triton X-100, the microcarriers showed light yellow, without cartilage morphology. The microcarriers became flocculous or like a hairbrush, and the area of contacting surface significant increased. After culture with cartilage cell for 2 hours, lots of spherical chondrocytes adhered to the microcarriers. HE stain of section confirmed that the celluar constituents of the specimens were removed, the specimens stained weakly positive for GAG, negatively for aggrecan, and positively for collagen type Ⅱ, respectively. Conclusion The detergent and trypsin can remove the cellular constituents and knock out the aggrecan from human articular cartilage while maintaining collagen type Ⅱ and GAG, and made the cartilage pieces flocculous or hairbrush-like. The chondrocytes can be well maintained in human articular cartilage derived microcarriers. Human articular cartilage derived microcarriers were prepared successfullly.

Female ; Humans ; Infant, Newborn ; Kidney Neoplasms ; Twins, Monozygotic ; Wilms Tumor

Objective To investigate the situation and change of antifungal resistance in clinical Candida and other fungal iso- lates from 5 hospitals in diverse geographic region of China.Methods Antimicrobial susceptibility testing of 8 000 fungat iso- lates collected during 2001 and 2005 were carried out with 25?g fluconazole disk and 1?g voriconazole disk using disk diffusion method as recommend by CLSI/NCCLS M44-A.Disk test plates were automatically read and results were recoded with the BIOMIC Image Analysis System.The equivalent MICs were automatically calculated by the BIOMIC System software.Results The proportion of Candida atbicans and non-Candida albicans (e.g.Candida glabrata) in the total fungal isolates did not change significantly from 2001 to 2005.The susceptibility rate of C.albicans to fluconazole and vorieonazole were stable during 2001 and 2005.However, the resistance to fluconazole and voriconazole increased variably in C.glabrata and other non-Can- dida albicans fungal isolates during the same period.Conclusions The voriconazole demonstrated higher activity against all yeast species in comparison with fluconazole.The increasing resistance to fluconazole and voriconazole in non C.albicans fungal isolates including C.glabruta suggests the importance of surveillance of fungal resistance in Candida isolates.

Objective To investigate the value of Cytochrome P450 (CYP3A5) * 3 gene polymorphism in providing individualized administration for the use of tacrolimus (Tac) in renal transplantation recipients.Method Pyrophosphate sequencing method was used to determine the CYP3A5 * 3 genotype of renal transplant patients in the first day after surgery.Sixty recipients were divided into experiment group and control group.Both groups of patients were routinely given the initial dose of Tac-4.0 mg/day in the first day after surgery.The experiment group of patients were given different doses of Tac based on the different CYP3A5 * 3 genotypes at the third day after surgery [for AA:0.12 mg/(kg· day),and for GG:0.06 mg/(kg· day)],and the control group of patients were given different dosages of Tac according to drug concentration.Different parameters were compared between two groups of patients:percentage of patients reaching the target concentration (3-8 μg/L) at the fifth day after surgery,days required to reach the target concentration level,times needed to adjust the dosage of Tac within two weeks.Result The percentage of patients reaching the target concentration in experiment group and control group was 90％ and 46.67％,respectively (P＜ 0.05).Days required to reach the target concentration were (3.67 ± 1.32) and (7.57 ± 3.42) on average,respectively (P ＜ 0.05).Times of adjusting the Tac dose in experiment group was significantly less than those in the control group (P＜0.05).In the experiment group,the target concentration was obtained even without dosage adjustment (70％).Conclusion Individualized adjustment of Tac doses for patients according to recipients' different CYP3A5 * 3 genotypes is beneficial for reaching target concentration as soon as possible,which is superior to traditional dosage regimen.

ObjectiveTo investigate the effects of intravenous (iv) remifentanil infusion on myocardial oxidative stress in rats.MethodsOne hundred and eighty male SD rats weighing 250-300 g were randomly divided into 15 groups (n =12 each):group control (group C); group ischemic preconditioning (group IPC); group remifentanil preconditioning ( group RPC ) ; while ia iv remifentanil infusion groups,iv remifentanil was infused at 4 different rates ( 1,5,10,20μg· kg- 1 · min- 1 ) and each rate was maintained for 15,60 and 120 min respectively.Myocardial ischemia was induced by occlusion of left coronary artery anterior descending branch for 30 min followed by 120 min reperfusion in 6 rats in each group.In group IPC myocardial ischemia was preceded by 3 cycles of 5min ischemia-5min reperfusion;whilein group RPC3cycles of 5min remifentanil infusion at 5 μg· kg-1 · min-1 were applied at 5 min interval before ischemia.Six rats in which I/R was produced were sacrificed in each group,myocardial infarct size (IS) and the area at risk (AAR) were measured and IS/AAR was calculated.The left 6 rats in each group were sacrificed at the corresponding time point (the end of each treatment)and superoxide radical expression and MDA and nitrotyrosine contents in myocardium were determined.Results IS/AAR was significantly decreased in groups IPC,RPC,1 μg·kg-1 ·min-1 × 120 min,5 μg·kg-1 ·min-1 × 60or 120 min and 10 μg· kg- 1 · min- 1 × 60 min as compared with group C.Compared with group C,the myocardial superoxide radical expression was significantly up-regulated in groups 1 μg· kg-1· min-1 × 120 min, 5 μg·kg-1 ·min-1 ×60 or 120 min,10μg·kg-1 ·min-1 ×60 or 120 min and 20 μg·kg-1 ·min-1 × 15,60 or 120min,and myocardial MDA and nitrotyrosine contents were significantly increased in group 20 μg· kg-1 · min-1 ×15,60 or 120 min.ConclusionLonger duration of high rate remifentanil infusion can induce myocardial oxidative stress in rats.