Objective To observe the protective effect of goserelin on ovarian function of premenopausal patients with breast cancer during chemotherapy. Methods Forty patients with breast cancer under 40 received adjuvant chemotherapy were randomly divided intotest group(20 cases) and control group (20 cases). Compared the recovery rate and time of menstruation between two groups. Results All the 40 patients finished the treatment. Recovery ratio of normal ovarian function in test group and control group was 75% and 50% (P =0.013) respectively, and the median menstrual recovery time in two groups was 4.65 months and 6.65 months (P = 0.046) respectively. Statistically significant differences were found between two groups. Conclusion Goserelin can effectively protect ovarian function during chemotherapy , increase the ratio of normal ovarian function, shorten menstrual recovery time, and shows good tolerance.

Objective To explore the intervention effects of trichostatin A ( TSA ) on specialization of rat bone marrow mesenchymal stem cells ( MSCs ) .Methods The rat MSCs were isolated, cultured and purified by the whole bone marrow adherent method in vitro, with morphological observation.The third generation of MSCs were selected, directional induced to osteoblasts, and divided into the TSA low dose group (0.1μmol/L), middle dose group (1μmol/L) , the high dose group (10μmol/L) according to different drug concentrations, seting up blank control group at the same time.MSCs proliferation and cell growth curve of each group were drawn by MMT, the activity of alkaliphosphatase ( ALP) was detected, and the levels of corebinding factor α1 (Cbfα1), basic fibroblast growth factor (bFGF) and insulin-like growth factors-1 (IGF-1) mRNA were detected by RT-PCR. Results The trend of MSCs growth curves in each groups were similar, compared with control group, the growth curve of TSA low dose group had no significant change, the TSA middle dose and high dose significantly promoted the proliferation of MSCs (P<0.05).Compared with control group, ALP activity of TSA low-, middle-and high-dose group were significantly higher at 4th,5th,6th(P<0.05).The expression levels of Cbfα1, bFGF and IGF-1mRNA were significantly higher than those of control group, respectively (P<0.05).Conclusion TSA can significantly promote the rat bone marrow mesenchymal stem cells to differentiate into osteoblast, which is possibly associated with up-regulation of Cbfα1, bFGF and IGF-1mRNA level.

Objective To investigate the effect of angiotensin 1-7(Ang 1-7) on renal proximal tubular epithelial cell(HK-2) transdifferentiation induced by high glucose.Methods All the raised HK-2 cells were divided into 5 groups: normal control group,high glucose group,high glucose with Ang1-7 group,high glucose with Ang1-7 and A779 group,high glucose with pioglitazone group.Expression of peroxisome proliferator activated receptor-γ(PPAR-γ) and α-smooth muscle actin(α-SMA) was detected by Western blotting,real-time PCR and immunofluorescence.Results The levels of PPAR-γ protein and mRNA in HK-2 cells were significantly increased after treatment with high glucose and Ang 1-7.Expression of α-SMA protein and mRNA was inhibited remarkably after treatment with high glucose and Ang 1-7.These effects of Ang 1-7 on HK-2 cells could be reversed by Mas receptor antagonist A779.Conclusion Ang 1-7 inhibits high glucose-induced expression of o-SMA in HK-2 cells,which is in part through the Mas.

Objective To investigate the effect of surfactant protein D(SP-D)overexpression on lipopolysaccharide(LPS)-induced monocyte chemoattractant protein-1(MCP-1)expression in human renal proximal tubular epithelial cells(HK-2)and its mechanism. Methods HK-2 cells were treated with LPS at various concentrations (0, 0.1, 1, 2, 5, 10 mg/L)for 8 h and at 5 mg/L for various time points(0, 2, 4, 8, 16, 24 h). Expression of SP-D was detected by Western blotting and real-time PCR. Expression of MCP-1 was determined by ELISA and real-time PCR. Human SP-D cDNA eukaryotic expression vector pEE14-hSP-D was transfected to HK-2 cells. The changes in transfected cells of SP-D protein were observed by Western blotting. Expression of MCP-1 was detected by ELJSA and real-time PCR. Results SP-D was expressed in HK-2 cells. The levels of SP-D protein and mRNA in HK-2 cells were significantly decreased after treatment with LPS(P<0.05). Expression of MCP-1 protein and mRNA was increased remarkably after treatment with LPS(P<0.05). HK-2 cells transfected with pEE14-hSP-D showed up-regulated expression of SP-D. The overexpression of SP-D inhibited the LPS-inducedexpression of MCP-1(P<0.01). Conclusions SP-D inhibits LPS-induced expression of MCP-1 in HK-2 cells. SP-D may play an important role in the modulation of renal inflammation.

Objective To evaluate the effect of aldosterone (Ald) on glomerular mesangial cells apoptosis and to explore the possible mechanisms.Methods Twenty-four Sprngue-Dawley rats were subcutaneously embedded with osmotic mini-pumps and randomly divided into 3 groups.Aldosterone (1.5 μg/h) was administrated subcutaneouly by osmotic mini-pumps in Ald group,eplerenone (Epl,100 mg·kg-1·d-1) and Ald (1.5 μg/h) was given to Epl group.And normal saline was used in control group (Con group).Systolic blood pressure and urinary albumin excretion rate (UAER) were detected on day 0,7,14,21,28.Blood and kidney samples were harvested on day 28.Plasma creatinine,potassium and aldosterone were measured.Renal paraffin sections were stained by PAS and the morphological changes were evaluated by light microscopy.Apoptosis index of mesangial cells were detected by TUNEL assay.The glomerular mesangial cells (MCs) were cultured in a DMEM-F12 media.MCs apoptosis was evaluated by staining cells with Annexin V and propidium iodide (PI) using flow cytometer.Expression of Bcl-2 and Bax mRNA was examined by RT-PCR.The protein level of Bad or phospho-Bad was measured by Western blotting.Results Ald-infused rats developed hyperaldosteronemia and hypokalemia.Rats in Ald group exhibited significant hypertension and marked albuminuria.Ald group rats showed increased number of TUNEL-positive mesangial cells when compared with control rats (P＜0.05).Aldosterone induced mesangial cells apoptosis in a time-dependent manner.Expression of Bcl-2 mRNA was decreased but Bax mRNA was increased in aldosterone treated MCs compared to that in Con group (P＜0.05).Aldosterone promoted dephosphorylation of cytosolic phospho-Bad compared with vehicle treated cells (P＜ 0.05).However,eplerenone attenuated these effects of aldosterone.Conclusion Aldosterone directly promotes mesangial cells apoptosis,and eplerenone can attenuate this effect of aldosterone.Dephosphorylation of cytosolic phospho-Bad may be the key role in the progression of mesangial cells apoptosis induced by aldosterone.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of C-terminal Src kinase (Csk) in Ang Ⅱ-infused rat model and cultured podocytes,and to explore the role of Csk in Ang Ⅱ-induced cytoskeletal rearrangement of podocytes.Methods Twenty-four Wista rats were randomly subjected to normal saline infusion,or Ang Ⅱ infusion at 400 ng · kg1 · min-1 (via subcutaneous osmotic minipumps) for 2 or 4 weeks.Renal histomorphology was evaluated through electron microscopy.The expression of glomerular Csk was analyzed by immunofluorescence and Western blotting.In vitro,conditionally immortalized mouse podocytes were cultured and treated with Ang Ⅱ doses ranging from 10-9 mol/L to 10-5 mol/L and for different hours.The expression of podocytes Csk was assessed by Western blotting.After transfection to podocytes with Csk siRNA,FITC-conjugated phalloidin was used to stain F-actin,to investigate the role of Csk in Ang Ⅱ-induced or cytochalasin D-induced cytoskeletal rearrangement.Results (1) Examination of Ang Ⅱ infusion rats glomerular and podocyte ultrastructure by electron microscopy revealed foot process effacement and fusion; (2) In Ang Ⅱ infusion rats,the expression of glomerular Csk was increased (P ＜ 0.05); (3) In vitro,Ang Ⅱ-stimuli up-regulated the expression of Csk (P ＜ 0.05),and the effects of Ang Ⅱ were on dose-dependent and time-dependent manner; (4) Ang Ⅱ-induced disruption of F-actin was alleviated by Csk siRNA transfection in cultured podocytes; furthermore,cytochalasin D depolymerized the F-actin cytoskeleton,while Csk siRNA stabilized the actin filaments.Conclusion The enhanced expression of Csk may be involved in Ang II-induced podocytes cytoskeletal rearrangement and foot process fusion.

Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.

[ ABSTRACT] AIM:To investigate the mechanism underlying breast cancer metastasis and to provide theoretical da-ta for studying the pathogenesis of breast cancer onset and development.METHODS: Human breast cancer MCF-7 cells were treated with different concentrations of furin inhibitorα1-PDX for 48 h.Wound healing assay and Transwell assay were applied to detect the migration and invasion abilities of the MCF-7 cells.The expression of cell migration-associated proteins, including membrane-type 1 matrix metalloproteinase ( MT1-MMP) , vascular endothelial growth factor ( VEGF)-C and VEGF-D, was determined by Western blotting.The protein levels of MMP2 and MMP9 in the supernatant were measured by ELISA. RESULTS:Compared with control group, 200 nmol/L of furin inhibitor exerted significant inhibitory effects on the cell mi-gration (P<0.05).The expression of cell migration-associated proteins MT1-MMP, VEGF-C and VEGF-D was significantly inhibited after treated withα1-PDX ( P<0.05 ) .Significant inhibitory effects of α1-PDX on the expression of MMP9 and MMP2 (P<0.05) in the supernatant were observed.CONCLUSION:Furin inhibitor suppresses the metastasis of MCF-7 cells via down-regulating the expression of MMPs and VEGFs.

OBJECTIVE To study effective and convenient method for paraffin oil sterilization.METHODS By using carrier qualitative germicidal test,to compare pressure steam sterilization,dry heat sterilization and cobalt-60(gamma)-ray radiation sterilization to test the sterilizing effect and operating procedure.RESULTS Pressure steam sterilization was unable to achieve 100% sterilizing effect,whether we extended the time or use the intermittent(sterilization).After dry heat or radiation sterilization processes,no microorganism was found.CONCLUSIONS Effect of sterilization with dry heat or radiation sterilization is trustable,but its packing,operation and equipment are requested strictly,and pressure steam sterilization may be not good for paraffin oil.

ObjectiveTo evaluate the effects of Ang Ⅱ on apoptosis of podocytes and explore the signaling pathwayof nephrin in preventingAng Ⅱ-inducedpodocyte apoptosis.MethodsDifferentiated mouse podocytes were exposed to Ang Ⅱ at different concentrations for 18 h or at 10-8 mol/L for variable incubation times.Undifferentiated mouse pedocytes were transfected using lipofectamine 2000 with the pcDNA3.1-mNPHS1 plasmid and stably transfected cell lines were generated with G418 selection.In separated experiments,untransfected mouse podocytes (MPC) and stably transfected podocytes with pcDNA3.1-neo and PcDNA3.1-mNPHS1 were exposed toAng Ⅱ(10-8 mol/L) or LY294002(a selective Akt inhibitor,50 μmol/L) for indicated times.Apoptosis was evaluated by flow cytometry.The expression of nephrin was assessed by quantitative real-time PCR,immunofluorescence and Western blotting.The phosphorylation level of Akt was determined by Westem blotting.Results(1) AngⅡ promoted podocyte apoptosis in a dose-and time-dependentmanner. PretreatmentwithlosartansignificantlypreventedAngⅡ -induced apoptosis. (2) Nephfin mRNA and protein were obviously decreased in podocytes exposed to 10-8 mol/L Ang Ⅱ for at least 12 h than those in vehicle-treated cells (P＜0.05).(3) Ang Ⅱ exposure for more than 15 min inhibited the phosphorylation of AKT in MPC,which was dramatically reversed by pcDNA3.1-mNPHS1 transfection,but not by pcDNA3.1-neo transfection. (4) Podocyte apoptosis was promoted byLY294002. Conversely,Ang Ⅱ-induced podocyteapoptosis was significantly alleviated by pcDNA3.1-mNPHS1 transfection.ConclusionAng Ⅱinduces mouse podocyte apoptosis which is suppressed by overexpression of nephrin through PI3K-Akt signaling pathway.