Objective: To investigate VEGF expression in NIH3T3 cells infected by adenovirus containing hVEGF165 gene and its influence on proliferation of NIH3T3 cells, and to observe the expression of hVEGF and its angiogenic effect in vivo. Methods: Adenoviral vector containing hVEGF165 gene was constructed and was used to infect NIH3T3 cells. The infection efficiency of adenovirus vector was examined by immunofluorescence and flow cytometry. Expression of VEGF in NIH3T3 cells and its levels in the culture medium were examined by immunohistochemical (IHC) staining, RT-PCR, and ELISA. The infected NIH3T3 cells were implanted in skin defect at rat back and the acellular dermis on the wound was obtained one week later; the expression of hVEGF was detected by IHC in the dermis and the density of vessels was determined under microscope. Results: NIH3T3 cells were effectively transfected by adenovirus containing VEGF gene in vitro, the transfection efficiency was in a dose-effect manner with multiplicities of infection (MOI) of the adenovirus. When MOI was 100, the infection efficiency was more than 95%. The expression of VEGF mRNA and protein was detected by RT-PCR and IHC 24 h after transfection. ELISA result showed that the high level of VEGF on the 3rd day after transfection and the level reached its peak 7 d after infection (1 052 pg/ml); VEGF expression was detectable 13 d after transfection. MTT assay demonstrated no significant difference in cellular proliferation between the transfection and non transfection group. Expression of hVEGF was also detected in vivo in mice, and the density of vessels in the experimental group was significantly higher than that in the control group (P<0.01). Conclusion: Adenoviral vector can effectively transfect VEGF gene into NIH3T3 cells; VEGF gene can be detected in vitro and in vivo; and it can promote neovascularization in the transplanted tissues.

Objective: To prepare NIH3T3 cells harboring microencapsulated VEGF gene and investigate the proliferation, activity and metabolic function of the modified cells. Methods: Microencapsulated VEGF modified NIH3T3 cells were prepared through an alginate-BaCl2 process. Morphological appearance of the microencapsulation and the cell morphology were observed under inverted phase microscope; untreated NIH3T3 cells served as control. The concentrations of VEGF in the culture supernatant (collected every 48 hours) were measured by ELISA; the proliferation and vitality of the cells were examined by MTT assay and flow cytometry with PI staining. Results: The microcapsules were round in shape and the cells grew well. There was no significant difference in the concentrations of VEGF,MTT values and vitalities of cells between the 2 groups. Conclusion: The growth and metabolic functions of NIH3T3 cells are not influenced by microencapsulated NIH3T3 cells harboring VEGF gene. The bio-properties of modified cells are similar to those of the control cells,which lays a foundation for transplantation of microencapsulated VEGF modified NIH3T3 cells in vivo.

Objective:To study the effects of glucosamine(GlcNH2) on immune function in mice.Method:The effects of GlcNH2 on murine proliferation of splenocytes were carried out in vitro.After feeding mice by GlcNH2,the phagocytotic functions of mononuclear macrophage,murine delayed type hypersensitivity(DTH) caused by sheep red blood cells(SRBC),the ability of antibody production(tested by HC50),and the index of immune organs(thymus and spleen) were deteimined in vivo.Results:GlcNH2 could promote the proliferation of splenocytes,phagocytotic functions of mononuclear macrophage,DTH,the ability of antibody production and the index of immune organs.Conclusion:Glucosamine can enhance immune function in mice such as cellular immunity,humoral immunity and non-specific immunity.

ObjectiveTo evaluate prognostic factors affecting local tumor progression after radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC).MethodsA total of 246 HCC patients (343 lesions) underwent RFA treatment in our department and were enrolled into this study.The average tumor size was 3.7 cm ( range 0.9 ～ 3.7 cm).Regular follow-up with enhanced CT was performed to evalutate the treatment results.Kaplan-Meier model and log-rank test were used in univariate analysis and COX regression model was used in multivariate analysis to identify risk factors for local tumor progression.ResultsThe local tumor progression rate was 11.4％ (39/343 lesions),and the average time from initial RFA to local tumor progression was 12.0 months.Univariate analysis indicated tumor size ( P ＜0.001 ),close to intrahepatic vessels ( P ＜0.001),tumor boundary ( P =0.020),pathological grade( P =0.010) and CEUS before RFA ( P =0.001) as risk factors for local progression.The following factors were identified as independent prognostic factors for local tumor progression by multivariate model:tumor size (P ＜ 0.001),isolated or close to intrahepatic vessels( P ＜0.001) and CEUS before RFA(P =0.018).ConclusionsTumor size,CEUS before RFA and close to intrahepatic vessels are the most important factors for local progression after RFA.Being awaring of possible risk factors for local tumor progression may increase the treatment efficacy and help to promote the use of RFA technique.

Microarray has become a popular biotechnology in biological and medical research.However, systematic and stochastic variabilities in microarray data are expected and unavoidable, resulting in the problem that the raw measurements have inherent "noise" within microarray experiments. Currently, logarithmic ratios are usually analyzed by various clustering methods directly, which may introduce bias interpretation in identifying groups of genes or samples. In this paper, a statistical method based on mixed model approaches was proposed for microarray data cluster analysis. The underlying rationale of this method is to partition the observed total gene expression level into various variations caused by different factors using an ANOVA model, and to predict the differential effects of G V (gene by variety)interaction using the adjusted unbiased prediction (AUP) method. The predicted G V interaction effects can then be used as the inputs of cluster analysis. We illustrated the application of our method with a gene expression dataset and elucidated the utility of our approach using an external validation.

This paper describes a calibration phantom system for QCT bone mineral density determination, which consists of 4-standard-solid-sample calibration phantom, a quality assurance (QA) phantom and the bone mineral density analysis software. The system adds to the new applications of CT systems, and provides a new method with a good accuracy and reliability for the examination, diagnosis, prevention, treatment of osteoporosis diseases and the observation of curative effect of drugs.
Absorptiometry, Photon ; instrumentation ; methods ; Algorithms ; Animals ; Bone Density ; Calibration ; Equipment Design ; Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Osteoporosis ; diagnostic imaging ; Phantoms, Imaging ; Software ; Tomography, X-Ray Computed ; instrumentation ; methods

12E7 Antigen ; Adolescent ; Adult ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; Brain Neoplasms ; secondary ; Cell Adhesion Molecules ; metabolism ; Child ; Child, Preschool ; Diagnosis, Differential ; Extremities ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Infant ; Ki-67 Antigen ; metabolism ; Male ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Oncogene Proteins, Fusion ; metabolism ; Repressor Proteins ; metabolism ; Sarcoma, Ewing ; metabolism ; pathology ; Sarcoma, Synovial ; diagnosis ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; Young Adult

Objective To discuss the feasibility of Shuyisha as hemostasis and repair material for liver wound. Methods Hemolysis rate, acute toxicity and eytotoxicity of Shuyisha were measured. A hemorrhage model was established by making an open wound (5 mm× 3 nun ×2 mm) on the left liver lobe of mice. Hemostasis was performed with Shuyisha in experimental group and with Surgicel in control group, when the hemostatic time and total blood loss (TBL) were accurately recorded and regular macro-scopic and histological observation carried out. Results The hemolysis rate of Shuyisha was 2.33%, with maximum tolerance does of over 0.48 g/kg and the eytotoxicity at zero. The hemostatie time of Shuy-isha was (5.00 ±0.00) s, with total blood loss of (0.88±0.18) g/kg, better than Surgicel (P< 0.05). Shuyisha was degraded completely within 14 days, with the wound healed within 21 days in ex-perimental group, much better than Surgieel. Conclusions The hemolysis rate, acute toxicity and cy-totoxicity of Shuyisha are up to the requirement of biomedical materials. Shuyisha has effective hemosta-sis, which may be related to its molecular structure and adhesion.

Objective： To study the changes of water molecular diffusion in the ischemic region by using MR dephase technique and discuss the potential mechanism of the diffusion changes at early stage. Methods: Totally 43 cases were studied retrospectively. There were 10 cases whose MRI examinations were performed within 6 hours，12 cases from 7－24 hours，7 cases from 2－7 days, 8 cases from 8－14 days, 6 cases from 15 days to 2 months. The apparent diffusion coefficients in the ischemic region were calculated. Results: The ADCav in the grey matter was 8.61×10－4mm2*s－1. The ADCav decreased to (4.72×10－4±1.51×10－4) mm2*s－1 in ischemic region at superacute stage, ADCav ratio to contralateral corresponding region was 0.55±0.18, and ADCav increased to (5.68×10－4±1.22×10－4) mm2*s－1 during the time range of 2-7 days, （9.22×10－4±2.07×10－4） during the time range of 8－14 days, and approaching （26.42×10－4+9.65×10－4） mm2*s－1 during the time range of 2 months. The pearson product- moment correlation between the changes of diffusion value and time was sighificent (r=0.95, P<0.001). ADCv increased at superacute stage and decreased over time. Conclusion: The diffusion of water molecules in ischemic region decreased at superacute stage, and the ADC increased over time. The anisotropy increased at superacute stage and decreased as the course developed. DWI could detect ischemic lesion much earlier than CT and routine MR examination. DWI has great value in the diagnosis of superacute stroke. The mechanism of the diffusion changes at early stage may be the intracellular toxicity edema.

Objective To evaluate the serum prostate specific antigen(PSA)change after I-125 radioactive seed implantation without androgen ablation therapy for prostate cancer,Methods A total of 13 patients with prostate cancer were included.The clinical stage was T_(1c)N_0M_0 in 8 cases,and T_(2a)N_0M_0 in 5. The Gleason scores were 5 in 4 cases and 6 in 9.The mean serum PSA was 8.2 ng/ml(range,2.8-14.6 ng/ml).I-125 seed implantation(D90=140-155 Gy)was performed.Serum PSA was monitored every month for the first 3 months and every 3 months for the next 20 months.Results The range of follow-up time was 3-23 months.At 1,2,3,6,9,12,15,18 and 21 months after implantation,the median serum PSA values were 6.7,5.0,2.7,1.6,1.2,0.9,0.8,0.8 and 0.7 ng/ml;and the median percentage change in ser- um PSA were 72%,51%,29%,20%,13%,11%,9%,9% and 8%,respectively.One month after brachy- therapy,30% of the patients had fluctuation in PSA.Conclusions Compared with radical prostatectomy and androgen ablation therapy,the rate of PSA decline is slower with fluctuation at early phase.The most sig- nificant decline of PSA occurs in the first 12 months,especially in the first 3 months after I-125 seed implan- tation.