Cloning and Expression Analysis of Squalene Epoxidase Gene in Poria cocos / 中国实验方剂学杂志

ObjectiveTo clone squalene epoxidase （SE）， a potential key rate-limiting enzyme involved in the synthesis pathway of Poria cocos triterpenes， from P. cocos and analyze for bioinformatics and expression. MethodThe total RNA was extracted by the kit and reverse-transcribed to cDNA. Specific primers were designed， and the cDNA was used as a template for cloning the SE gene， which was analyzed for bioinformatics. The expression of P. cocos qualene epoxidase（PcSE） was examined by Real-time polymerase chain reaction（Real-time PCR） in P. coco Shenzhou No. 10， Xiangjing 28， and 5.78 strains. ResultThe full length of PcSE is 1 571 bp， containing four exons and three introns. The obtained CDS sequence is 1 413 bp， encoding 470 amino acids. This protein is a hydrophobic protein with no signal peptide structure and has two transmembrane structural domains with a FAD/NAD （P） binding domain and SE structural domain localized to the mitochondrial membrane and the plasma membrane. The homologous sequence alignment with fungi of the Poriferae family is 80.92%， and the phylogenetic tree shows that PcSE protein is most closely related to P. cocos from the US. The results of Real-time PCR showed that the PcSE was expressed in all three strains， with the highest expression in 5.78 strain， and there was no significant difference in PcSE expression among the three strains. ConclusionFor the first time， the PcSE gene was cloned and analyzed from P. cocos， providing a basis for further research on the function of PcSE and the analysis of P. cocos triterpene biosynthesis pathway.

The effectiveness of virtual reality therapy in relieving upper limb hemiparesis after stroke:A meta-analysis of randomized and controlled trials / 中华物理医学与康复杂志

Objective To analyze the clinical efficacy of using virtual reality ( VR) to improve the upper limb function of hemiplegic persons. Methods A search was conducted in the PubMed, Cochrane Library, EM-BASE, CNKI and Wanfang Data as well as VIP for reports of randomized and controlled studies of using VR in train-ing upper limb function after stroke. A meta-analysis was then performed using version 5. 3 of the Review Manager software. Results Ninety studies involving 879 patients were found and analyzed. The data showed that VR was sig-nificantly more effective than conventional training in improving Fugl-Meyer assessment scores. It was not superior, however, in improving average Functional Independence Measure scores or performance in the box and blocks test. Conclusion VR is superior to conventional training in promoting the recovery of upper limb function after a stroke.

A virtual visualization system for colon polyp inspection / 中国医疗器械杂志

The virtual colonoscopy, virtual flattening and virtual splitting method are enhanced by the GPGPU model. The novel virtual eversion method is integrated for fast polyp detection. The experimental result showed that the system and various visualization methods can represent the colon inner-surface clearly and exactly, supporting real-time man-machine interaction. The proposed system is promising in human gastrointestinal cancer and polyp inspection.

Subcutaneous implantation of tissue-engineered heart valve leaflets in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the viability of tissue-engineered heart valve leaflets prepared with cell-polymer constructs in nude mice.</p><p><b>METHODS</b>Sheep endothelial cells and smooth muscle cells/fibroblasts were seeded on patches of PHA and implanted subcutaneously in athymic mice (BALB/C). The cell-polymer constructs were harvested 12, 14, 21 and 28 days after implantation.</p><p><b>RESULTS</b>Fourteen days after implantation, the cell-polymer constructs exhibited similar color with the autologous tissues, and HE staining showed more numerous cells in the implant. At 28 days following implantation, muscular fibers were formed in the cell-polymer constructs. V-G staining showed positive collagen staining in the implant at 12 days after implantation, while the control implants retrieved 28 days after implantation did not show extensive tissue formation or muscular fiber formation.</p><p><b>CONCLUSION</b>The cell-polymer constructs can survive in vivo and has the potential to grow into autologous valve leaflets in the nude mice.</p>

An improved method for the isolation of rat hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To present an improved method to obtain pure, viable, freshly isolated hepatic stellate cells.</p><p><b>METHODS</b>Adult male SD rats were used. All procedures were performed with the animals under sodium pentobarbital anesthesia. Three days after the single intravenous administration of 1 ml liposome-encapsulated CL2MDP, which has selective cytotoxicity of Kupffer cells, livers were perfused with D-Hank's solution containing 100 U/ml heparin for 10 to 15 minutes, and then with 0.05% collagenase dissolved in D-Hank's solution for 25 to 30 minutes. The liver was then gently homogenized and further incubated in 0.025% collagenase, and 0.005% DNAase I for 30 minutes at 37 degrees C under constant stirring. This suspension was filtered through stainless steel gauze and centrifuged for 2 minutes at 50 x g to remove parenchymal cells. Sinusoidal cells in the supernatant were recovered by centrifugation for 10 minutes at 300 x g. The cells were resuspended in the presence of 28.7% Nycodenz stock solution. The final concentration of Nycodenz at this stage was 11.5%. Following centrifugation for 17 minutes at 1400 x g, The cells at the top of this Nycodenz solution were collected. Cells were resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, The cells were seeded in 50 ml culture flask at a density of 500,000 cells/ml, The cell viability was determined by trypan blue exclusion staining, the purity of hepatic stellate cells was identified by the expression of Desmin using immunocytochemistry method. Endogenous peroxidase staining was used to detect Kupffer cells.</p><p><b>RESULTS</b>The yield rate of hepatic stellate cells was 3 x 10(7) per rat, the cell viability was more than 95%, the desmin positive cell rate was 90%, no endogenous peroxidase positive cells were detected.</p><p><b>CONCLUSION</b>The method for the isolation of hepatic stellate cells was developed without Kupffer cells confusion. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.</p>

Effect of basic fibroblast growth factor and hyaluronic acid on proliferation of rabbit chondrocytes in vitro / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro.</p><p><b>METHODS</b>Chondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetra-zolium bromide. (MTT, Sigma, M2128).</p><p><b>RESULTS</b>Basic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50-500 ng/ml and that of HA was 10-50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone.</p><p><b>CONCLUSIONS</b>bFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.</p>