The Wnt pathway plays a crucial role in skeletal development and is indispensable for determination of OS cell lines. In recent years, experimental evidences on Wnt signaling pathway in OS cell lines and OS animal models, and that of Wnt signaling pathway as a diagnosis and prognosis marker of OS suggested that Wnt signaling pathway plays an important role in the progression, invasion and metastasis of OS. In addition, the strategy and safety evaluation to target Wnt to treat OS are underway. Exploring the significant role of Wnt signaling pathway in OS may aid in personalizing therapeutics to increase patient survival.

T, p.R394W in exon 9.

Objective This study has explored the role of antibody against annexin A2 in patients with antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE). Methods Using purified recombinant annexin A2, IgG anti-annexin A2 antibody was measured by ELISA in 101 APS patients, 41 SLE patients with thrombosis, 124 SLE patients without thrombosis and 120 healthy controls. Results The positive rate of IgG anti-annexin A2 antibody in APS patients and SLE patients with thrombosis was 21.8%, 26.8%, respectively, they were all significantly higher than in SLE patients without thrombosis (6.5%). IgG anti-annexin A2 antibody was associated with thrombosis and/or pregnancy morbidity (P<0.01). Conclusion Anti-annexin A2 antibody is associated with thrombosis and/or pregnancy mnrbidity. It suggests that anti-annexin A2 antibody may be helpful in identifying in some potential AIRS.

Euphorbia kansui (EK) is a toxic herbal drug, and often used after vinegar-processing to reduce its toxicity. In present study, a 1H-NMR based metabonomic approach was used to evaluate the detoxification effect of vinegar-processed EK. The water extracts of EK and VEK were administered orally to male SD rats at doses of 9 g x kg(-1) x d(-1) for 1 week, respectively, and one more week observation was further conducted. The control group was orally given with saline. Histopathological studies of liver samples on the 8th and 15th day were conducted, and the metabolites of rat urine and liver were analysed by 1H-NMR. Histopathological studies of liver samples from EK and VEK treated rats showed no negative impacts. In metabonomic analyses of urines, changes of metabolites indicated liver damages, kidney lesions and imbalance of gut microbes in the second week. VEK-treated rats showed a quite lower toxicity compared with EK-treated ones. The present study revealed that the metabonomic approach might be helpful for the evaluation of toxicity of EK and detoxic effect of VEK.
Acetic Acid ; chemistry ; Animals ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; toxicity ; Euphorbia ; chemistry ; Inactivation, Metabolic ; Liver ; drug effects ; metabolism ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; methods ; Rats ; Rats, Sprague-Dawley ; Urinalysis

OBJECTIVE To investigate the role of several xenobiotic metabolism-associated CYP450 isoforms in the auto-activation of hepatic stellate cells(HSCs) invitro. METHODS HSCs were isolated from adult Wistar rats and cultured on plastic as an in vitroauto-activation model. Positive expression of α-smooth muscle actin(α-SMA)was used as an activated marker of HSCs and detected by immunocytochemical staining in HSCs cultured for 1,2,5 or 11 d. The expressions of CYP450 isoforms were analyzed by real-time RT-PCR in the HSCs. RESULTS The immunocytochemical stai-ning showed no α-SMA expression in HSCs cultured for 1 d,while its expression gradually increased during culture since day 2. ln terms of α-SMA expression,HSCs cultured for 1,2,5,and 11 d were classified as the quiescent,early,middle and later stages of activation,respectively. The RT-PCR results revealed that CYP1B1,CYP2B1/ 2,and CYP2E1 mRNA were expressed at high levels in the early stage of HSCs activation(at day 2),which were 2.1-,1.6-,and 23.9-fold those in the quiescent HSCs(day 1),respectively. Further study revealed that mRNA expressions of these up-regulated CYPs in the early stage of activation were diminished at the subsequent two stages. The levels of CYP1A1 and CYP1A2 mRNA decreased constantly throughtout the activation process. CONCLUSION Multiple xenobi-otic metabolism-associated CYP450 isoforms might be involved in the auto-activation of rat HSCs invitro.

AIM: To investigate the cell cycle arrest ind uced by hypoxia, hypoxia inducible factor-1 and their possible mechanism in huma n ovarian cancer cell line SW626. METHODS: CoCl 2, a chemical inducer of hypoxia and hypoxic cell culture chamber were used to induce chemical and physical hypoxia in human ovar ian cancer cell line SW626. The method of ‘decoy’ was used to block the functi on of HIF-1? because it acts as the core sequence of the target gene as a compe titor combined to the HIF-1?. The cells were divided into group A1 (normal oxyg en), A2 (normal oxygen plus HIF-1? decoy), B1 (CoCl 2), B2 (CoCl 2 plus HIF-1 ? decoy), C1 (hypoxia) and C2 (hypoxia plus HIF-1?). The expression of the HIF -1? protein, mRNA and cell cycle analysis were detected by Western blotting, RT -PCR and flow cytometry (FCM). RESULTS: The expression level of HIF-1? protein in group B1 (3 .75?1.31) and group C1 (3.48?1.01) was significantly higher than that in g roup A1 (0.97?0.31) (P0.05). FCM showed that the G 0/ G 1 phase was markedly increased in group B1 (81.78?24.33) and group C1 (77 .62?22.76) and was significantly higher than that in group A1 (49.49?18.54 ) (P0.05). CONCLUSION: Both CoCl 2 and physical hypoxia could distinctly i nduce cell cycle arrest in G 0/G 1 phase and the expression of HIF-1? in huma n ovarian cancer cell line SW626. HIF-1? plays an important role in cell cycle arrest induced by hypoxia in human ovarian cancer cell line SW626.

Objective To optimize the formulation of a new kind of ultrasound contrast agents carrying the sensitizer of hematoporphyrin(HP)with[Poly(lactic-co-glycolic acid),PLGA]for material.Methods The technique of double emulsion was applied to produce HP loaded PLGA ultrasound microbubbles,which was optimized through orthogonal test using encapsulation efficiency for the detected index.Then morphology and distribution of HP-PLGA microbubbles were observed through light microscope and scaning electron microscope.The size,Zeta potential and the properties of releasing behavior and ultrasound imaging in vitro of Hp-PLGA contrast agents were detected.Results The optimization parameters were picked out as 10 mg/ml for concentration of HP,40 mg for PLGA,and 1/5 for volume ratio of water inside to dichloromethane.The optimized HP-PLGA contrast agents were spheric with the mean size of 602.3 nm,and Zeta potentiaI of-(17.1±1.6)mV.The drug loading and encapsulation efficiency of HP-PLGA were(2.15±0.15)%and(63.5±2.6)%,respectively.And the releasing behavior of HP-PLGA contrast agents in vitro was that after an obvious release of about 35.1%in former 24 h,there were 86.5%HP-PLGA released within 14 days.The ultrasound imaging of HP-PLGA could be enhanced obviously in vitro.Conclusions The self-made HP-PLGA ultrasound microbubble might be a useful tool for delivering sensitizer and thus provide a novel strategy for sonodynamic therapy on tumor.

The ultrasonographic appearances of osteosarcomas and the roles of ultrasonography in the diagnosis and surgical staging of osteosarcomas were investigated. A comparative study was performed on 45 cases of osteosarcomas by ultrasonography and radiography. Bony changes, periosteal reaction and soft tissue mass were evaluated for each lesion. The results showed that ultrasonography revealed a solid mass around bone in 42 patients, bone destruction in 24 patients and periosteal reaction in 16 patients. Plain radiographs showed bony changes in 44 patients and no bony change in remaining one patient, shadowing of soft tissue swelling in 30 patients, and pulmonary metastases in 3 patients. Surgical biopsy and pathological examination confirmed osteosarcoma in all 45 patients. Soft tissue mass was confirmed in 42 patients surgically. The diagnostic accuracy of soft tissue masses by ultrasonography and radiography was 100 % (42/42) and 71.4 % (30/42), respectively. The positive rate of ultrasonography and radiography in displaying bony changes was 53.3 % (24/45) and 97.8 % (44/45), respectively. In conclusion, in the detection of soft tissue mass of osteosarcoma, ultrasonography is superior to radiography, and in displaying bony changes of osteosarcomas, radiography is superior to ultrasonography. So it may come to a conclusion that plain radiography combined with ultrasonography can completely display the bony and soft tissue lesion of osteosarcomas.

Background The pathological foundation of diabetic retinopathy is the breakdown of the bloodretina barrier induced by multifactors.Objective This study was to investigate the retinal morphologic change and the levels of serum vascular endothelial growth factor(VEGF),endothelin(ET)and nitric oxide(NO)in diabetic rats.Methods Forty healthy female SD rats were randomly divided into experimental group and control group and 20 rats for each group.Acute diabetes models were established by the intraperitoneal injection of 60 mg/kg streptozotocin in experimental group and the equal volume of buffer solution was injected at the same way in the control group.The serum VEGF level,ET level and NO concentration of diabetic and control rats were detected using ELISA double antibody sandwich method,125I radioimmune method and nitrate reduction method respectively at 2,4,6 and 8 weeks after injection.The eyeballs of rats were enucleated at the eighth week for retinal pathologic examination.This experiment followed the Measures for the administration of experimental animals of Shanghai City.Results The retinal structure was normal throughout the experimental duration in the control group.However,retinal edema and cellular disorganization appeared at 4 months and retinal blooding could be seen with the extending of diabetes course.The levels of serum VEGF and ET in each experimental group were significantly higher than those in control groups(P＜0.05).The levels of serum NO elevated in 2-month experimental group compared with same-phase control group(Z =-2.193,P＜0.05),and those in 6-and 8-month experimental groups were significantly lower than in corresponding control groups(Z =-2.449,Z =-2.236,P＜0.05).With the progression of the disease,the levels of VEGF and ET increased gradually,but the levels of NO decreased gradually,showing statistically significant differences(P＜0.05).The level of serum VEGF showed a positive correlation with serum ET level and a negative correlation with the serum NO concentration(r=-0.814,r=-0.803,P＜0.01)in the experimental group.A negative leaner relation was also found between serum ET level and serum NO concentration(r=0.821,P＜0.01).Conclusions The serum VEGF,ET and NO levels are closely associated with the degree of retinal lesion in early diabetic models.These results suggest that serum VEGF,ET and NO levels may be the important indexes predicting the course of retinal disease in diabetic rats.

Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P<0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P>0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P<0.01). The expression of RANKL was inhibited (P<0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P>0.05), but the expression of RANKL was higher in PMMA group than in control group (P<0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P<0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P<0.01) and the expression of OPG mRNA was significantly increased (P<0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P<0.05), but the expression of OPG mRNA had no significant difference between them (P>0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.