BACKGROUND:Studies have reported that puerarin can reduce bone resorption and promote bone formation. OBJECTIVE: To further explore the effects of puerarin at different doses on the proliferation of osteoblasts culturedin vitro. METHODS: Osteoblasts isolated from the cranium of newborn rats were culturedin vitro, and then passage 3 cels were cultured in DMEM medium with the presence of 0, 20, 40, 80 and 100 μmol/L puerarin+10% fetal bovine serum for 48 hours. The viability of osteoblasts, alkaline phosphatase activity and mineral node formation were determined using MTT, alkaline phosphatase kit and alizarin red staining, respectively. RESULTS AND CONCLUSION:The viability of osteoblasts treated with puerarin at either 40 or 80 μmol/L was significantly higher than that of the control group. Alkaline phosphatase activity and the number of mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 μmol/L when compared with the untreated cels. These findings demonstrate that puerarin is able to promote osteoblast proliferation in vitro.

BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfacted into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-βl (TGFβ_1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytas were isolated by use of trypsin digestion method. Vector was PLNCX_2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed_2 double digestion, to build PLNCX_2-RFP. TGFβ_1 gene was amplified from the PGEMT-TGF and connected with PLNCX_2-RFP following double digestion, to build PLNCX_2-TGFβ_1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX_2 group and transfected PLNCX_2-TGFβ_1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supematant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ_1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX_2-TGFβ_1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ_1 and RFP, which showed that the eukaryotic expression vector PLNCX_2-TGFβ_1-RFP had been successfully built as expectation. They were then transfected into packaging calls and cultured, the virus titer was defined as 1×10~6 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent calls formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the calls proliferated actively. NO concentration in the transfected PLNCX_2-TGFβ_1-RFP group was higher than that of transfected PLNCX_2 group (P < 0.05), no difference was significant between control group and transfected PLNCX_2 group. The control group and the group transfected PLNCX_2 showed no TGFβ_1 expression, while TGFβ_1 concentration was (28.08±3.73) ng/L in the transfected PLNCX_2-TGFβ_1-RFP group. PLNCX_2 ratroviral vector-mediated human TGFβ_1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage calls proliferate actively.

Objective To evaluate the efficacy and safety of the transobturator tension-free vaginal tape (TVT-O) surgery, a minimally invasive surgery for treating patients with the stress urinary incontinence (SUI). Methods 28 cases with female stress uri-nary incontinence treated by TVT-O procedure from April 2006 to June 2008 were retrospectively analyzed. The follow-up time is from 4 to 24 months. Results The mean operation time was 23 min (rang 10～30 min) and the mean intraoperative bleeding was 24 ml (range 15～40 ml). An indwelling catheter had been used for 3 d because of urinary retention in 3 cases. 24 patients(85.7%)reached complete control after the surgery,effective control in 4 patients(14.35%). Conclusions TVT-O surgery is an effective, safe, minimally invasive management to treat the stress urinary incontinence.

ObjectiveTo identify differential genes of malignant melanoma using gene chip expression profiles.MethodsAgilent Human 1A OligoDNA was employed to find out difference in gene expression between malignant melanoma and nevus. The total RNA was isolated from two type tissues, labled the fluorescent to the probe, hybridized, washed and analyzed.ResultsAmong the 21073 target genes, 1596 genes were differentially expressed in malignant melanoma, including 733 genes up-regulated, and 863 down-regulated.ConclusionThe gene chip technique can screen genes that may be specifically expressed in malignant melanoma.