BACKGROUND: Tissue-engineered materials containing osteoblasts have been widely used in the treatment of orthopedic diseases. How to improve hypoxic tolerance and the biological behavior of osteoblasts under hypoxic conditions is an important goal of the research on tissue engineering.OBJECTIVE: To explore the effect of M2 macrophage supernatant and Eucommia flavonoids on the biological behaviorof osteoblasts under hypoxia to pave ways for tissue engineering. METHODS:After MC3T3E1 cells were resuscitated, and M2 macrophages were isolated, induced and cultured, all cells were divided into control, M2 macrophage, Eucommia flavonoids and combination groups. The control group received no intervention. Cells in the M2 macrophage, Eucommia flavonoids and combination groups were exposed to M2 macrophages or/and 100 mg/L Eucommia flavonoids, respectively. RESULTS AND CONCLUSION:The cell viability in the M2 macrophage, Eucommia flavonoids and combination groups was significantly higher than that in the control group. The percentage of cells in the S and G2/M phases in the combination group was significantly higher than that in the control group. The relative mRNA expression levels of Runx2, collagen type Ⅰ and alkaline phosphatase, alkaline phosphatase activity and relative concentration of calcium ions in the combination group were significantly higher than those in the other three groups. To conclude, M2 macrophage supernatant combined with Eucommia flavonoids can promote osteoblast proliferation and differentiation under hypoxic conditions, maintain normal mineralization and osteogenesis of osteoblasts, and improve the tolerance ability of osteoblasts to hypoxia.

Blood Vessels ; injuries ; Extremities ; blood supply ; injuries ; Humans ; Ligation ; Pulmonary Embolism ; etiology ; prevention & control ; Vascular Surgical Procedures ; methods ; Venous Thrombosis ; etiology ; prevention & control

Objective To study the effect of ursolic acid(UA) on apoptosis of the ovarian cancer cell line SKOV3 and its action mechanisms. Methods The SKOV3 cells were treated with UA at different concentration. The proliferation inhibition was examined by MTT assay. Morphological examination and flow cytometry were used to detect apoptosis. RT-PCR and immunocytochemical method were used to detect the expression of wild type p53,an apoptosis related genes. The semi-quantification of protein expression was analyzed by pathological image-analysis. Results UA inhibited the proliferation of SKOV3 cells strongly. Apoptosis of SKOV3 cells were induced by UA treatment. The morphology of SKOV3 showed changes such as chromatin aggregation, nuclear and cytoplasmic condensation and apoptotie bodies appearance. UA caused significant G0/G1 arrest with a concomitant decrease of cell population in S and G2/M phases. Apoptotic cells were found by flow cytometry. The maximal early apoptotic rate was (28.7±2.4)% when exposed to 40 μmol/L for 48 h. The expression of p53 gene and protein was enhanced. All these effects were in a dose and time-dependent manner Conclusion UA induced apoptotic processes in the ovarian cancer SKOV3 cell line were the mechanism involving p53 pathway.

Humans ; Korea ; Medicine, Chinese Traditional

OBJECTIVE: To evaluate the effect of Eucommiae ulmoides on the microstructural and ultrastructural change of femur and lumber vertebrae in ovariectomized rats.METHODS: 80 4-month-old female SD rats were randomized to 4 groups: sham operation group,model group,alfacalcidol group,E.ulmoides group.In the sham operation group,the abdominal cavities of the rats were cut open to expose ovaries before being sutured.However,the rats in the other 3 groups were ovariectomized bilaterally.At 10 days after operation the rats were administered intragastrically with corresponding drug.After 3 months,femur and lumber vertebrae were dissected and fixed in 4% glutaral for 3 days,then decalcified in 4% EDTA for 4 weeks,and then the structure of femur and lumber vertebrae was observed under light microscope and transmission electron microscope(TEM).RESULTS: Observed under light microscope,the femur and lumber vertebrae were full of fat in the ovariectomized rats and in which the density of bone trabecula decreased significantly;however,in alfacalcidol-treated rats and E.ulmoides-treated rats,the amount of fat decreased markedly while the density and thickness of bone trabecula increased.Observed under TEM,in the ovariectomized rats,rough endoplasmic reticulum(RER),cytolysosome and mitochondrial were plentiful inside osteoclasts in femur and lumber vertebrae,partial osteoblasts atrophied,the amount of RER and other cellular organ decreased significantly and apoptosis appeared for partial cells;however,in alfacalcidol-treated group and E.ulmoicdes-treated group,the above changes improved significantly.CONCLUSION:E.ulmoides has remarkable beneficial effect on osteoporosis in ovariectomized rats.The mechanism might be attributed to its inhibition on the synthesis of fat and the activity of osteoclasts,promotion on the activity of osteoblasts and inhibition on the apoptosis.

Objective To investigate the morphological alteration and the protein expression of caspase -3 in long -term denervated posterior cricoarytenoid muscles (PCAMs)in order to find the appropriate time point of rein‐neration in long -term denervated PCAMs .Methods A total of 45 patients with vocal paralysis were recruited and devided into 3 groups ,the 3~6 months denervation group ,the 6~12 months denervation group ,the 1~2 year denervation group .12 adults served as control group .The morphological alteration was evaluated using HE staining and the change in expression of caspase -3 ,an apoptosis related factor ,were observed using immunehistochemistry stain and western blot .Results With elongation of denervation time ,there were increased denaturation in the mus‐cle fibers .The nucleus moved inside and some of them concentrated .Caspase-3 showed weak staining in innerva‐ted ,however ,by 3-6 months of muscle denervation there was a significant accumulation of caspase -3 protein in myofibers ,6~12 months and 1~2 years of denervation ,expression of caspase -3 protein in myofibers was de‐creased significantly .In western blot ,the change in protein expression of caspase -3 was observed an 21-fold (P< 0 .01) increase from 3~6 months denervated muscles to innervated muscles ,11-fold (P< 0 .01) increase from 6~12 months denervated muscles to innervated muscles ,3~fold (P< 0 .01) increase from 1~2 year denervated muscles to innervated muscles .Conclusion The morphological alterations and changes in expression of caspase -3 indicated there was a high amplitude of apoptosis in denervated posterior cricoarytenoid muscles within 1 year .

Tumor microenvironment plays a crucial role in neoplastic evolution process as it may provide functional interactions between neoplastic and non-neoplastic cells involved in tumor invasion,metastasis and angiogenesis,which implies that it can be regarded as a novel target for the antitumor therapy of polysaccharides.This review focuses on the protumor variations existing in tumor microenvironment,such as growth factors,immunosuppressive factors,proteases,glycoproteins,etc.,which may offer possible mechanisms of polysaccharides on the regulation of their corresponding functions.

Humans ; Lower Extremity ; blood supply ; May-Thurner Syndrome ; etiology ; Venous Thrombosis ; etiology

Six taxane diterpenoids were isolated from the stem bark of Tax us cuspidata Sieb. et Zucc.. On the basis of physical and chemical properties and spectra analysis,they were identified as 2'-de sacetoxyaustrospicatine(Ⅰ). taxol(Ⅱ),cephalomannine (Ⅲ), 2-deacetoxyl taxinine J (Ⅳ),taxinine B(Ⅴ) and 10-deacetyl-7-0-?-Dxylosyl-taxol (Ⅵ).

Great and extensive progress has been made in joint surgery in the last decade. With the evolution of surgical concepts and the development of operational technique and equipment, indications, selective range for surgica l operation and management in the period of perioperation have been surveyed in new perspectives and with deeper insights. This article introduces the situation of joint surgery of China in these years, briefly reviews the latest progresses in the fields of the arthroscopy, the arthroplasty and the joint cartilage repa ration, and thus suggests developmental strategies for Chinese joint surgery in the future.