Osteoprotegerin(OPG) ,receptoractivatorofNF -?Bligand (RANKL)andreceptoracti vatorofNF -??(RANK)areimportantmoleculesthatregulateosteoclastogenesis .OPGisdecoyreceptorof RANKL .ThebindingofOPGandRANKLblocksconnectingofRANKLandRANK ,inhibitsdevelopmentof osteoclast.ThebindingofRANKLandRANKinducesaserieskinasecascadereaction ,activatestranscription factors,introducestheproliferationanddifferentiationofosteoclastprecursors .Severalhormonesandos teotropicfactorsregulatetheformationandfunctionofosteoclastthroughtheabovemolecules .

BACKGROUND: Whehter estradiol (E2) can change the expression of peroxisome proliferator-activated receptor gamma-2(PPAR-γ2) during differentiation of bone marrow stroma cells should be studied further.OBJECTIVE: To investigate the effects of 17β-estradiol (E2) on the gene expression of PPAR-γ2 mRNA and PPAR-γ2 protein.DESIGN: Contrast observational trial.SETTING: Laboratory Department of People's Hospital of Deyang City; Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech Limited Company.MATERTALS: A 3-month-old female SD rat (200±20) g used to isolate bone marrow stromal cells was obtained from Animal center of Chengdu Chinese Medicine University (Medical Experimental Animal Number 11). Dulbecco's mimimum essential medium (DMEM) was obtained from Hyctone Compnay. 1α, 25(OH)2D3, dexamethasome (DEX) and E2 were purchased from Sigma Company. Total RNA kits were obtained from Omga. One step RNA PCR kit (AMV) was obtained from Takara Shuzo Co, Ltd. Northern direct HRP labeling and detection kit was purchased from PIERCE. Western blotting luminol reagent was obtained from Santa cruz.METHODS: The experiment was carried out from April 2001 to July 2002 in the Laboratory Department of People's Hospital of Deyang City, Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech (0, 0.1, 10, 1 000 n) was used to interfere cell differentiation for 3 days. Cultured cells were crushed with Tris-Triton X-100 PBS; activity of alkaline phosphatase was detected with Beckman CX-7 biochemical analytical device; effect of 100 g/L formalin, stained with Weigert-hematoxylin for 10 minutes, rinsed with water, differentiated with 5 g/L hydrochloric ethanol, stained with Van Gieson, desiccated with ethanol of the fractional volume of 0.95, cleared with dimethylbenzene with RT-PCR, Northern blot and Western blot during cell differentiation.protein.proliferated within 24-72 hours. Cells shaped as triangle, multiple angles and fusiform. Three days later, volume of adherent cells was increased and colony, and 10 days later, they confluenced. Bone marrow stroma cells in many generalight red to yellow). Red plasma presented synthesis of collagen. The deeper the red was, the more the collagens were.pression of PPAR-γ2 mRNA was (4.0±0.4)%, (1.7±0.2)% and (2.8±0.2)% (t=6.1, 7.2, 11.5, P＜ 0.01), which was higher crease the expression of PPAR-γ2 protein. When concentration of E2 was 0.1, 10 and 1 000 nmol/L, expression of PPAR-γ2 protein was (2.2±0.2)%, (2.6±0.2)% and (4.1 ±0.2)%, which was higher than that in 0 nmol/L E2 group [(1.2±0.10)%, t=6.6, 8.5, 13.2, P＜0.01].CONCLUSTON: E2 can inhibit expression of alkaline phosphatase and promote differentiation of bone marrow stromal cells and expression of PPAR-γ2 mRNA and PPAR-γ2 protein.

Objective To investigate the effects of 17?-estradiol (E2) on the gene expression of typeⅠA bone morphogenetic protein receptor (BMPR-ⅠA) and core-binding factor alpha 1 (Cbf?1) in rat bone marrow stromal cells exposed to the differentiation medium and to elucidate the effects of E2 on osteoblastogenesis. Methods Adherent bone marrow stromal cells were cultured in differentiation medium containing DEX (10 -7 mol/L), 1,25-(OH)2D3 (10 -9 mol/L) and different concentrations of E2. Effects of different concentrations of E2 on the gene expression of BMPR-ⅠA and Cbf?1 was quantified by RT-PCR based on the comparison with an internal reference, ?-actin expression, and identified by Northern blotting. Alkaline phosphatase (ALP) activity of cells was detected. Contents of type Ⅰ collagen were determined by Van Gieson staining. Results E2 could evidently inhibit the expression of BMPR-ⅠA and Cbf?1 mRNA during the differentiation process of bone marrow stromal cells into osteoblasts in a dose-dependent manner. These were confirmed by Northern blotting. The ALP activity increased in a concentration-dependent manner, but the amount of type Ⅰ collagen decreased in a concentration-dependent manner. Conclusion E2 can significantly inhibit the gene expression of BMPR-ⅠA and Cbf?1 in bone marrow stromal cells and inhibit osteoblastogenesis in vitro.

Aim To investigate the effects of tamoxifen on the proliferation and DNA synthesis of cultured human pituitary adenoma cells and make a further investigation of the mechanism of the inhibitory effect of tamoxifen on the proliferation of pituitary adenoma cells. Methods The techniques of MTT colorimetry, 3H-TdR, flow cytometry, PKC activity detection and cAMP/ cGMP levels detection were used to detect or observe the effects of tamoxifen on proliferation, DNA synthesis, cell cycle, PKC activity and cAMP/ cGMP levels of cultured human pituitary adenoma cells, respectively.Results ①Tamoxifen (0.1,1 and 10 ?mol?L -1) inhibited the proliferation and DNA synthesis of cultured human pituitary adenoma cells in a dose-dependent manner.② tamoxifen (1,10 and 20 ?mol?L -1) increased the ratio of G_1 phase of pituitary adenoma cells, and decreased the ratio of S and G_2 phase markedly;②compared with control, PMA, a PKC activator, increased the activity of membrane and total PKC in human pituitary adenoma cells. However, after a 15-min treatment with tamoxifen (10 ?mol?L -1),a significant reduction of the activity of cytoplasm, membrane and total PKC in human pituitary adenoma cells was observed;③tamoxifen (1 and 10 ?mol?L -1) increased the amount of cAMP in the cytoplasm of human pituitary adenoma cells, but had no effect on that of cGMP. Conclusion These data provide an important clue to explore the molecular mechanisms of the inhibitory effect of tamoxifen on the proliferation of pituitary adenoma cells, and suggest that the modulating effect of tamoxifen on the proliferation of pituitary adenoma cells results from interactions of several cellular signaling pathways.

AIM:To investigate the effects of 17?-estradiol (E_2) on the gene expression of typeⅠA and typeⅠB bone morphogenetic protein receptor (BMPR-ⅠA,ⅠB) in rat bone marrow stromal cells exposured to the differentiation medium and to elucidate the effects of E_2 on osteoblastogenesis. METHODS: Adherent bone marrow stromal cells were cultured in differentiation medium containing DEX(10 -7 mol?L -1 ) and 1,25(OH)_2D_3 (10 -9 mol?L -1 ) and different concentrations of E_2. The gene expression of BMPR-ⅠA,ⅠB was quantified by semiquantitative RT-PCR. RESULTS: E_2 evidently inhibited the expression of BMPR-ⅠA mRNA in bone marrow stromal cells.The suppression was dose-dependent. When examined under various concentrations of E_2 (0-10 -6 mol?L -1 ),the expression of BMPR-ⅠA mRNA were decreased from (25.7?2.5)% to(16.3?1.5)%( P

Objective: To explore the effect of exercise preconditioning (EP) on pathological cardiac hypertrophy and heart failure (HF) in pressure over-loaded experimental rats.
Methods:A total of 60 SD rats at the age of 6 weeks were randomly divided into 3 groups, n=20 in each group. Sham-operation group, Transverse aortic constriction (TAC) group and EP + TAC group. The cardiac function and structure were evaluated by echocardiography, patholgical changes and HF biomarkers were examined for EP effect at 4 and 8 weeks after TAC.
Results:Compared with Sham-operation group, the cardiac function and structure had obvious changes in the other 2 groups. Compared with TAC group, the ejection fraction in EP+ TAC group increased 15%, the heart weight index and left ventricular weight index decrease 15.7%and 20%respectively at 8 weeks after TAC, all P<0.05. Compared with Sham-operation group, the mRNA and protein expressions of ANP and BNP increased in TAC group at 4 and 8 weeks after TAC, increased in EP+TAC group at 8 week after TAC. Compared with TAC group, the mRNA expressions of ANP and BNP in EP+TAC group decreased 47%and 62%at 4 weeks after TAC, decreased 44%and 28.1%at 8 weeks after TAC, all P<0.05;the protein expression of ANP and BNP in EP+TAC group decreased 22.3%and 48%at 4 weeks after TAC, decreased 21.5%and 38.3%at 8 weeks after TAC, all P<0.01.
Conclusion: EP may improve cardiac pathological hypertrophy in pressure over-loaded rats at the early stage, and delay the heart failure process.

Objective Three-dimension (3D) cell matrix adhesion in vivo is fundamentally important for a wide variety of cellular physiological and pathological phenomena, however, the cell-matrix 3D adhesion is hardly observed in vitro. We present the human foreskin fibroblasts (HFF) formed 3D adhesion complexes on the thoroughly acellular human amniotic matrix (TAHAM). Methods TAHAM were produced by suspending digestion with trypsin. The HFF were seeded on 6 well plate, matrigel and TAHAM individually.The light microscope, scanning electronic microscope, immunohistochemistry and immunofluorescence were used to observe the micro-structures and detect the type Ⅰ , Ⅲ, Ⅳ, Ⅵ collagen, laminin, fibronectin, TGF-β1, TGF-β2, FGF of the TAHAM. Phase contrast microscope was engaged to observe the morphology of HFF. The time-lapse CCD and the trace analysis software were employed to prescribe the cell migration. The 3D adhesion foci were identified by the laser confocal microscope. The strain of the TAHAM was tested by the universal mechanical testing instrument. Results The fibers of the TAHAM were intact, type Ⅰ , Ⅲ,Ⅳ, Ⅵ collagen, laminin, fibronectin were positive, TGF-β1, TGF-β2, FGF were negative. HFF had a bipolar extension to form multilayer cell clusters networks and grew into the matrix. All of the seeded cells survived three weeks under regular culture without transfer. On TAHAM, HFF moved in a straight line with a speed of 12 μm/h. α5 integrin (green), paxillin (red) and fibronectin (blue) co localized to form 3D adhesion complexes (white). Conclusion The main molecular components and biomechanical properties is preserved in TAHAM. HFF forms 3D adhesion complexes on TAHAM. Cell morphology and migration of HFF on TAHAM correspond to that under 3D adhesion behavior.

Objective To investigate the P2X3 receptor expression in L6-S1 dorsal root ganglion (DRG)and bladder detrusor in a rat model of neurogenic bladder and urethra. Methods Eighty Sprague-Dawley rats wererecruited and randomly divided into a sacral injury group, a suprasacral injury group and a control group. Spinal tran-section was performed to establish the animal model of neurogenic bladder and urethra in rats of the sacral injurygroup and suprasacral injury group. Check the P2X3 receptor expression in DBG and bladder detrusor among thethree groups by Western blot test at 20 days after model establishment. Results P2X3 receptor expression in L6-S1DRG of sacral injury group was significantly less than that of the suprasacral injury group, which was in turn signifi-cantly higher than that of the control group. P2X3 receptor expression in bladder detrusor of sacral injury group wassignificantly lower than that of the suprasacral injury group, which was in turn significantly higher than that of thecontrol group. Conclusion There was close relationship between P2X3 receptor expression and dysfunction of blad-der and urethra.

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis, treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage I b1, two stage I b2, one stage IIa, one stage IIb, and one stage IIIb. On the basis of their stages of condition, one subject with stage III b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively. It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis,treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage Ⅰ bl, two stage Ⅰ b2,one stage Ⅱ a, one stage Ⅱ b, and one stage Ⅲb. On the basis of their stages of condition, one subject with stage Ⅲ b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively.It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.