Objective To establish an effective method of isolating and culturing circulating fibrocytes from human peripheral blood and study the relationship between the expression specific molecule markers expression and the morphological characteristics. Methods Total peripheral blood leukocytes were isolated from human peripheral blood by being centrifuged over Ficoll-Paque and cultured in DMEM supplemented with 20% fetal calf serum.Adhered cells were detected with immunocytochemistry,FCM and electron microscopy.The collagen synthesis was studied by measurement of the hydroxyproline concentration in the medium with chemical method. Results After 9 days cultue in vitro,CD34,CD45 and collagen Ⅰ staining was positive and 83.5% of these cells secreted collagen Ⅰ detected by FCM.Electron microscopy of circulating fibrocytes showed morphological characteristics of fibroblasts.The hydroxyproline concentration in the medium was 11.17%mg/L,which was statistically and significantly different compared with 8.07mg/L in the control medium(P

BACKGROUND: Ovariectomized animals usually are employed for the study of women osteoporosis while little is known about theenergy balance of ovariectomized animals. Many previous studies showed that soy isoflavone could decrease the hyperlipemia resulted by high-fat feed, and how about the effect of soy isoflavone on the energy metabolism in ovariectomized animals?OBJECTIVE: To explore the pathologic effect of soy extract and its active components on the energy metabolism in ovariectomized rats so as to provide adequate evidences for the primary rehabilitation and prevention of type Ⅱ diabetes mellitus, disturbance of lipid metabolism as well as hypertension and coronary atherosclerotic heart disease.DESIGN: A randomized and controlled trial with the experimental animals as subjects.SETTING: Laboratory of Cell and Biochemistry in a university.MATERIALS: Ninety Wistar rats (SFP grade, license code: scxk11-00-006) were randomized into 9 groups: normal group, sham group,model group, estrogen group, high-dose soy flavone group, low-dose soy flavone group, high-dose soy extract group, low-dose soy extract group and soy polysaccharide group, with 10 rats in each group.INTERVENTION: Except the rats in normal group and sham group,the bilateral ovaries of other rats were all removed. From one week after operation, body weight and daily food intake were detected once a week.Six weeks later, the rats were killed to calculate the forage transformation efficiency, measure body length, work out the body mass index (BMI),and separate the abdominal fat and weight.MAIN OUTCOME MEASURES: ①Effect of soy extract on the abdominal fat accumulation of ovariectomized rats. ②Effect of soy extract on food intake of ovariectomized rats. ③Effect of soy extract on forage transformation efficiency of ovariectomized rats.RESULTS: After ovariectomy, the food intake, body weight and BMI all raised, and the forage transformation efficiency increased, with abdominal fat accumulation. The changes of energy metabolism induced by ovariectomy were all weakened at different degrees in estrogen group, soy extract group and soy flavone group, while no influenced was found in soy polysaccharide group.CONCLUSION: The ovariectomized rats can be used as animal model of the climacteric fat women. The soy extract, with the effective component of flavonoid, can reduce the pathologic changes such as the increase of food intake, body weight and forage transformation efficiency induced by ovariectomy.

Aim To investigate the phytoestrogenic effects and mechanism on ER?,ER? expression levels at the protein of quercetin and psoralen in T47D cells.Methods T47D cells were treated with 10 ?mol?L-1 quercetin and 10 ?mol?L-1 psoralen for 48h respectively,total cell extracts examined for the levels of ER? and ER? protein by RT-PCR and Western blot analysis.The estrogen-like effect of quercetin and psoralen and their relations with the estrogen-receptor were evaluated by pure estrogen receptor antagonist ICI182,780 as a tool.Results Psoralen(10 ?mol?L-1) and quercetin(10 ?mol?L-1) up-regulated the expression of ER? mRNA and protein,without any effect on ER? mRNA and protein levels.The above up-regulation on ER? protein could be inhibited by adding estrogen receptor antagonism ICI182,780.Conclusion Psoralen and quercetin exerts estrogenic activity to ER-positive cells proliferation through interaction with ER? expression.

Objective To investigate the effect of puerarin on proliferation of human embro fibroblast and extracelluar matrix. Method Human embro fibroblasts were incubated with 0～400 ?g/mL puerarin for 24～96 h. The MTT method was used to assay the biological activities and extracelluar matrix formation in different time and different dose of pueratin. The cell cycle distribution was detected by flow cytometric analysis. Result Incubation cells in presence of puerarin of 40～400 ?g/mL for 24～72 h could significant inhibit cell proliferation and restrain the collagen synthesis in a dose- and time-dependent manner (P

Objective To observe the therapeutic effects of curcumin on pulmonar y fibrosis rats induced by bleomycin. Method One hundred and forty-four male Sp rague-Dawley rats were randomly divided into 6 groups (24 rats in each group). Rats in the model control group,positive medicine group,and high-,moderate-and low-dosage curcumin groups were injected with a single dose of bleomycin th rough trachea,and rats in sham-model control group with the same volume of nor mal saline. One day after the injection,curcumin solution of different dosages (200,100,50 mg?kg-1?d-1) was respectively given to rats in the high-,mod erate-and low-dosage curcumin groups daily by gastric gavage,while the same v olume of normal saline was given to those in the sham-model control group and m odel control group,and the same volume of prednisone(0.56 mg?kg-1?d-1) was given to those in the positive medicine control group. On the 7th,14th,and 28t h day after medication,8 rats in per treatment group were randomly killed,the changes of lung function were observed,and the lung was incised to make patholo gical sections which were stained with HE and Mallory. Results Curcumin improved lung function. The results of pathological examination showed that curcumin rel ieved the pathogenic changes of the lungs of pulmonary fibrosis rats induced by bleomycin. Conclusion Curcumin exerts a certain curative effect on pulmonary fib rosis rats induced by bleomycin.

Objective To explore the possible functional mechanism of Er-xian Decoction and its component herbs on the expression of extrogen receptor(ER) in adrenal development rats.Methods Fifty-four development female SD rats were randomly divided into nine groups:normal control group(A),positive control group(B),Er-xian Decoction group(C),Herba Epimedii group(D),Rhizoma Curculiginis group(E),Radix Morindae Officinalis group(F),Radix Angelicae Sinensis group(G),Cortex Phellodendri group(H),Rhizoma Anemarrhenae group(I).After the rats were administrated for six days,serum were got to measure testosterone(T).Bilateral adrenal gland was weighed and embedded by paraffin.Mean optical density(MOD) were studied by the immunohistochemical method and image analyzing system.Results The adrenal coefficient was increased in group B and C,and the level of T was decreased in group C,D,E and I.ER? was mainly observed in the cytoplasm of zona glomerulosa and zona fasciculate.MOD in group C was increased while declined in group B.ER? was mainly observed in nucleus of zona glomerulosa and zona fasciculate.MOD in group C,D,E and I was less than group B.Conclusion Er-xian Decoction and its component herbs of warming Shen(Herba Epimedii,Rhizoma Curculiginis) and nourishing Yin(Rhizoma Anemarrhenae) play a phytoestrogen role by influencing the expression of ER? and ER ? in the adrenal cortex.

Objective To study the effect of Fufangbiejiaruanganpian (FFBJRGP) on deposition of collagens around hepatic Disse space in experimental alcohol liver fibrosis (ALF). Methods The ALF rat model was induced by feeding a mixture of alcoholic liquid which included alcohol, vegetable oil and pyrazole, combined with a high fatty food. Then, all of the ALF rats were treated with FFBJRGP in high, moderate, and low dose. At the time of termination of the treatment, HYP level in liver tissue, collagen Ⅳ and LN levels in the serum and pathological changes were studied by Mallory stain and sirius red stain. Results The levels of HYP in hepatic tissues, collagen Ⅳ and LN in sera are increased remarkably in ALF rats, while all these indexes were reduced after moderate or low-dose FFBJRGP treatment (P

Estradiol ; blood ; Female ; Hot Flashes ; prevention & control ; Humans ; Isoflavones ; therapeutic use ; Luteinizing Hormone ; blood ; Middle Aged ; Phytotherapy ; Postmenopause ; blood ; drug effects ; Prospective Studies ; Soybean Proteins ; therapeutic use ; Soybeans ; chemistry ; Sweating ; drug effects ; Syndrome

Aim To explore the effects of tanshinone IIA on cell proliferation via G protein-coupled estrogen receptor inductive and regulative pathway in typical es-trogen receptor and G protein-coupled estrogen receptor positive T47D breast cancer cells. Methods The pro-liferation rate of T47 D cells influenced by tanshinone IIA was analyzed by MTT assay. G protein-coupled es-trogen receptor agonist G1 and GPER antagonist G15 were employed as tools. GPER SiRNA was applied to build GPER gene silence T47D cells. GPER expres-sion influenced by tanshinone IIA was measured by Western blot. Results The proliferation rates of T47D cells treated with 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol· L-1 of tanshinone IIA were decreased significantly. Such effects could be attenuated by G1 or enhanced by G15 . Growth of GPER SiRNA transfected T47 D cells were significantly inhibited by 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol·L-1 of tanshinone IIA treating. Result of Western blot showed that tanshinone IIA at 1 × 10 -5 mol· L-1 and 1 × 10 -6 mol · L-1 could induce de-crease of GPER protein expression in T47D cells. Conclusions Tanshinone IIA shows inhibitory effects on proliferation rate of T47 D breast cancer cells via GPER pathway. Tanshinone IIA could perform regula-tive function on GPER expression level in target cells.

Aim To investigate the effects of quercetin and psoralen on the proliferation of human breast carcinoma cells in vitro.Methods Effects of quercetin and psoralen on the cell proliferation was tested in ER-positive MCF-7 cells by flow cytometer and Western blot.And the estrogen-like effect of psoralen and its relation with the estrogen-receptor were evaluated by pure estrogen receptor antagonist ICI182,780 as a tool.Results ① Psoralen(10 ?mol?L-1) and quercetin(10 ?mol?L-1) stimulated proliferation of MCF-7 cells compared with vehicle control,and the cell cycle was impulsed from G1 to S,DNA synthesizing was enhanced.It was also found the above function on boosting MCF-7 cell proliferation could be inhibited by adding estrogen receptor antagonism ICI182,780.② Psoralen(10 ?mol?L-1) and quercetin(10 ?mol?L-1) up-regulated ER? protein levels without altering ER? levels.The above up-regulation on ER? protein could be inhibited by adding estrogen receptor antagonism ICI182,780.Conclusions Psoralen and quercetin have the estrogen-like activities through the estrogen response pathway.And they exert estrogenic activity to MCF-7 cell proliferation through interaction with ER? expression.