Objective To establish a new animal model of chronic cervical compressive myelopathy and to assess its feasibility. Methods Eighteen Chongming goats were divided into two groups: control group (n=3) and experimental group (n=15). The balloon was placed into the C3 intervertebral space by anterior approach operation, and the syringe valve was fixed subcutaneously. Contrast agent was injected percutaneously into the valve (0.1 ml/week) to inflate the balloon progressively to produce chronic compression. In the control group, the balloon compression system was placed and immediately removed; percutaneous puncture was performed each week without injecting anything. The Tarlov scores were assessed in each group every four weeks. The goats underwent X-ray, CT and MRI under general anesthesia every four weeks. The spinal cord specimens were pathologically examined at test level at the end of experiment. Results The Tarlov scores were 5 (normal) at all time points in the control group. Tarlov scores were not changed in the experimental group four weeks after surgery (n=13); at eight weeks after surgery (n=11) the Tarlov scores were 4 in 2 goats and 5 in 9 goats; and at twelve weeks after surgery (n=9) the Tarlov scores were 2 in 3 goats, 3 in 4 goats and 4 in 2 goats. The balloon compression system was stable in the experimental group. Radiological findings showed that the cervical spinal cord compressed progressively in the experimental group as time went by, and those in the control group underwent no noticeable change. Pathological examination showed neuronatrophy, increased gap around the neurons, mild demyelinated and vacuolar degeneration in the experimental group at eight weeks after surgery, and these changes were deteriorated twelve weeks after surgery. There were no noticeable pathological changes in the control group and four weeks after surgery in the experimental group. Conclusion The postoperative behavior, radiological and pathological findings of the animals consist with the character of chronic cervical compressive myelopathy, indicating that the balloon compression system in the present study can be used to establish a reliable and stable animal model of chronic cervical spinal cord compression.

Objective To evaluate the influence of interferon adjuvant therapy on immune parameters in postoperative patients with localized clear cell renal cell carcinoma (LCCRCC) and explore the related clinical significance. Methods Thirty-five patients with LCCRCC were treated with interferon α-2b hypodermic injection after surgery (6 MIU/time, three times per week for three months). Immune parameters, including CD4+,CD8+,CD4+/CD8+,CD16+56+,CD19+, IL-2, IL-6, IL-10, IL-8, and TNF-α, were determined before and at the 1st, 2nd, 4th, 8th, and 16th week after therapy. And the results were compared before and after therapy. ResultsThree months after therapy, the levels of CD4+, CD8+ and CD4+/CD8+ were not significantly different from those before therapy. The of CD16+56+ was increased significantly during the first two weeks' of treatment (P<0.05) and was significantly declined at the end of therapy (P<0.01). Compared with that before therapy, CD19+ levels were decreased in the 1st, 2nd and 4th week after treatment (all P<0.01), and was significantly increased at the 16th week (P<0.01). The level of IL-8 was significantly decreased at the 4th week after therapy (P<0.05) and TNF-α level was increased at the 8th week after therapy (P<0.01); the levels of other humoral immune parameters were not significantly different from those before therapy. Conclusion Treatment with interferon α-2b hypodermic injection (6 MIU/time, three times/week for three months) has a limited effect on promoting the immunity of patients with LCCRCC, and its influence on the long-term survival patients also needs further study.

Objective To characterize the expression of lipid metabolism-related genes in the fatty liver of obese ob/ob mice. Methods Four 18-weeks old male ob/ob and 4 control mice were sacrificed after 16 h fasting. Their body mass, ratio of liver wet mass to body mass and liver triglyceride contents were examined. H-E staining and Oil red O staining were performed to observe the histological changes and lipid deposition of the liver. Real-time quantitative RT-PCR was used to detect mRNA expression of lipid metabolism-related genes in the liver. Results The body mass, ratio of liver wet mass to body mass and liver triglyceride contents were significantly higher in ob/ob mice than those in control mice (P<0. 05). H-E staining and Oil red O staining showed severe hepatic steatosis in the ob/ob mice. The mRNA levels of fatty acid translocate (CD36), fatty acid binding protein 1 (FABP1), fatty acid synthase (FASN), acetyl-CoA carboxylase 1(ACC 1), elongation of very long chain fatty acids family member 6 (ELOVL6) and stearoyl-CoA desaturase 1 (SCD1) were significantly higher in ob/ob mice than those in control mice (P<0. 05); and there were no significant differences in the mRNA levels of peroxisome proliferator-activated receptor a(PPARa), palmitoyl-CoA oxidase (ACOX1), apoprotein B (ApoB) or microsomal triglyceride ttansfer protein (MTP) between the two groups (P> 0. 05). Conclusion The genes closely related to fatty acid uptake and de novo fatty acid synthesis are up-regulated in ob/ob liver, and those related to fatty acid oxidation and lipid transportation and VLDL secretion are not greatly affected at mRNA level.

Hypoxia-induced erythropoietin signaling plays an important role in tumor growth and invasion. In the present study, we investigated the contribution of erythropoietin signaling pathway to castration-resistant prostate cancer and the development of a neuroendocrine phenotype. Immunohistochemical staining showed that the erythropoietin and erythropoietin receptor scores in castration-resistant prostate cancer and androgen-dependent prostate cancer were 7.55 versus 4.5 and 7.45 versus 5.9,respectively (P < 0.001). Furthermore, a cell proliferation assay was conducted, and the differential expression of erythropoietin and erythropoietin receptor in LNCaP cells and hypoxia-induced LNCaP cells was evaluated using western blot and quantitative real-time PCR. The proliferation capacity of hypoxia-induced LNCaP cells was similar in cultures of both fetal bovine serum and charcoal-stripped fetal bovine serum, suggesting that LNCaP cells acquired hypoxia-induced androgen-independent growth. After 2 weeks of hypoxic culture, LNCaP cells showed a neuroendocrine cell change and increased expression of neuron-specific enolase, erythropoietin, and erythropoietin receptor; knockdown of erythropoietin receptor reversed the hypoxia-induced upregulation of neuron-specific enolase in the LNCaP cells. In conclusion, the concurrent upregulation of erythropoietin and erythropoietin receptor in castration-resistant prostate cancer suggests that the erythropoietin/erythropoietin receptor autocrine loop plays an important role in the progression of castration resistance and is responsible for the development of a neuroendocrine phenotype.

Objective To prepare a novel collagen scaffold material using Basa fish (Pangasisus haniltoa) skin as the ingredient and to analyze the structural characteristics, physical properties and degradability of the prepared material, so as to explore whether Basa fish can replace terrestrial mammals for preparing a novel collagen scaffold material. Methods Basa fish skins were lyophilized to obtain the membrane material after repeated degreasing, decolorization and dedoping. Crude protein content was determined by the Kjeldahl method. Structure of the materials and its pore size and distribution were analyzed by scanning electron microscopy (SEM). Porosity was measured by the liquid displacement technique, and tensile strength was tested using universal testing machine. The changes of viscosity with temperatures were detected to determine the denaturation temperature of the material.The material was immersed in the phosphate-buffered solution (0.1 mol/L, pH 7.4), which was placed in a constant temperature shaker at 37°, and the water absorption and weight loss rates of the material were detected. Results The crude protein content of the collagen scaffold material was 95.2%, with visually uniform thickness. SEM photographs showed that one side of the material had a rough surface and porous structure, on which varying sizes of pores distributed uniformly; the other side was smooth with dense pores. The porosity of the material was (55.50±1.94)%, thickness was (0.66±0.10) mm and tensile strength was (18.82±0.94) MPa. The denaturation temperature of the material was 34° before thermo-crosslinking and 36° after thermo-crosslinking. The water absorption of the material was (379.77±77.81)% at 48 h. At 28 d after thermo-crosslinking, the degradation rate was (80.22±2.49)%, and the pH value of buffer was 6.67±0.05. Conclusion The collagen scaffold material from Basa fish skin can be made into the biological membrane with uniform thickness, and the membrane comprises double structures: dense layer and loose layer. This material exhibits excellent mechanical strength and appropriate denaturation temperature, but its degradation is fast, which needs further improvement.

Prostate cancer is the most common malignant tumor in men of western countries and its incidence is increasing in China. However, the traditional methods of screening and diagnosing of prostate cancer are of limited value. Multiparametric magnetic resonance imaging (MRI) is an examination which combines morphologic sequences with one or more functional sequences. Multiparametric MRI can not only display the anatomical structures and morphologic changes of organs, but also reflect some histological components, providing pathophysiological and biochemical information of tissues and guiding prostate targeted biopsy at the same time. These advantages give multiparametric MRI high application value in the diagnosis and evaluation of prostate diseases. This review summarized the recent progress in multiparametric MRI diagnosis of prostate cancer.

Objective To establish a novel transgenic mouse model of human PRKAG2 cardiac syndrome that overexpresses a PRKAG2G100S mutation, so as to lay a foundation for further studying the role of human PRKAG2 gene in the development, morphology, and function of mouse heart. Methods Human PRKAG2 with G100S mutation was sub-cloned into a multiple cloning site located in the downstream of α-myosin heavy chain (a-MHC) promoter of the plasmid. After the construction of the transgenic expressing vector, C57BL/6J mice were selected as the genetic background, and the transgenic mouse model of PRKAG2-G100S mutation was buitt by microinjection. Genotype was further confirmed using specific primer PCR. Real time PCR and Western blotting analysis were used to examin the expression of human PAKAG2(G100S) mRNA and protein, respectively. Results Two strains of transgenic mice were successfully developed using backcross breeding, which specifically overexpressed the human PRKAG2-G100S mutation in the cardiac tissues of F2 generations by the methods qPCR and Western blotting at both mRNA and protein levels. Moreover, the PRKAG2-G100S mutation was successfully passed steadily. Conclusion We have successfully established a human PRKAG2-G100S transgenic mouse model, which can help to further explore the role of PRKAG2-G100S mutation in the development and function of mouse cardiac tissue in the PRKAG2- G100S cardiac syndrome.

Objective To establish a cardiomyocyte model over-expressing mutant human PRKAG2 by infecting neonatal SD rat myocardial cells with constructed recombinant adenovirus vector Ad-PRKAG2 (R302Q)-IRES2-EGFP. Methods PRKAG2 (R302Q)-IRES2-EGFP was directly cloned into entry vector pDONR221 by using Invitrogen GatewayTM technology. Then BP and LR recombination reactions yielded the recombinant adenovirus vector containing human PRKAG2 (R302Q) gene. The pAd-PRKAG2 (R302Q)-IRES2-EGFP was digested by Pac , and transfected into 293 cells. After packaging, amplification and purification, the virus was used to infect neonatal rat cardiomyocytes. Then the expression of PRKAG2 protein was assayed by Western blotting analysis in the infected neonatal SD rat cardiomyocytes. Results Restriction enzyme digestion analysis and the sequence analysis confirmed that PRKAG2(R302Q) gene was successfully inserted into the adenovirus vector. The myocardial cells infected with Ad-PRKAG2(R302Q)-IRES2-EGFP gave off strikingly bright green fluorescence and PRKAG2 protein was proven significantly over-expressed by Western blotting analysis (P<0.05). Conclusion The recombinant adenovirus containing human PRKAG2(R302Q) gene has been successfully constructed and expressed in neonatal rat cardiomyocytes, which paves a way for further study of PRKAG2 (R302Q) gene mutation.

Objective To investigate the effects of Tet-on gene regulated HIF-1?expression on cell apoptosis of hepatoma cells in vitro. Methods The apoptosis of hepatocellular carcinoma cell lines HePG2 was measured after HIF-1?expression was activated in HePG2 in vitro by Tet-on expression system. Results The amplified products were confirmed as the cDNA of HIF-1?by DNA sequencing, and obtained pTHE-HIF-1?was verified by endonuclease digestion. The HePG2Tet-on cells express HIF-1?constantly. After being incubated under different concentrations of doxycycline for 48 h, HIF-1?gene could inhibit HePG2 cell apoptosis. The apoptosis rates were 59. 6%、50. 9%、38. 1%、30. 5%、23. 9%、18. 3% respectively when the concentrations of doxycycline were 0?g/ml,0. 02?g/ml,0. 2?g/ml、1?g/ml、2?g/ml,5?g/ml (P

Objective To understand the infection status and variation tendercy of Enterobius vermicularis infection among children at national monitoring spots of soil-transmitted nematodosis from 2006 to 2010,and master the epidemic regularity,so as to provide the evidence for making control strategy and evaluating the control effect. Methods A total of 22 national monitor-ing spots of soil-transmitted nematodosis were established according to the National Surveillance Program of Soil-Transmitted Ne-matodiasis(Trial),and the children aged 3-12 years were examined through adhesive cellophane anal swabs,then the infec-tion rates of children with different ages,genders,nationalities and education levels were analyzed. In addition,the advantage, disadvantage,opportunity and threat of the monitoring work were analyzed by SWOT analysis. Results A total of 17 068 chil-dren were examined in 22 monitoring spots from 2006 to 2010,and 1 363 of them were found being infected with E. vermicu-laris,the average infection rate was 7.99%,and the infection rates of male and female children were 7.39%and 8.70%,respec-tivel;the average infection rates in each year were 10.01%,9.68%,7.41%,6.96%and 6.57%,respectively. From 2006 to 2009,the infection rates of E. vermicularis in children in Fujian Province was the highest,which were 56.15%,53.42%, 37.82%and 49.53%,respectively,but in 2010,the infection rate in Guangdong Province(46.06%)was the highest. The fur-ther analysis demonstrated that the female children,3-6 age group,Li nationality and children at kindergarten stage had relative-ly high infection rates. The SWOT analysis showed that the advantage of E. vermicularis monitoring in China was its wide cover-age and continuity,and the disadvantage was the relatively small investment from the government,the opportunity was that the national monitoring spot could drive the monitoring work at the provincial,county and other levels,and the threat was that the work was paid less and less attention to in recent years. Conclusion Though the infection rate of E. vermicularis in children at national monitoring spots of soil-transmitted nematodosis has been decreased year by year,high-endemic areas still exist,and thus the work on enterobiasis control and prevention still needs to be strengthened.