Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.

ObjectiveTo investigate the method of establishing damaged endometrial stromal cells (ESC) model in vitro.Methods ( 1 ) From June to December 2011 ESC from normal endometrim at proliferation phase ( n =8 ) and secretory phase ( n =8 ) were isolated,cultured and identified in vitro.( 2 ) ESC was treated with different concentrations of mifepristone or withdrawal of mifepristone at different time point.The proliferation inhibition percent was measured by cell counting kit-8 ( CCK-8 ). ( 3 ) 0 μmol/L (control group)and 60 μmol/L(experimental group) concentration of mifepristone was added into ESC for 48 hours,then withdrew of mifepristone,continued to be cultured for 48 hours.The morphological changes were observed and apoptosis of ESC in different menstrual cycle were detected by flow cytometry.The mRNA and protein level of vascular endothelial growth factor ( VEGF),caspase-3,8,and 9 were determined by one-step quantitative real-time PCR (Q-PCR) and western blot.Results( 1 ) ESC from 16 specimens of endometrium were all isolated and cultured successfully. (2) The proliferation inhibition rate of ESC was correlated with concentration and duration of mifepristone positively. The proliferation of ESC could be recovered at a range of time after withdrawal of mifepristone.However,when the concentration of mifepristone was 100 μmol/L,the growth of ESC recovered very hardly. (3) The damaged ESC spacing increased,the spindle shape and vacuolization in the cytoplasm were observed in experimental group; the rate of apoptosis of these damaged cells was significantly increased compared with control groups,which were (52 ± 12)％ vs.( 13 ± 5 ) ％ at the proliferative phase and (53 ± 6) ％ vs.( 32 ± 3 ) ％ at the secretory phase ( all P ＜0.05).The relative mRNA level of VEGF was 0.52± 0.12 in experimental group and 1.00 ± 0.17 in control group at proliferation phase (P ＜0.05).And the relative mRNA level of VEGF was 0.19 ±0.03 in experimental group and 0.81 ±0.07 in control group at secretory phase (P ＜ 0.05).The relative level of VEGF protein in the experimental group were both decreased 1.98 and 2.79 folds at the proliferation phase and the secretory phase when compared with those in control group,respectively ( P ＜ 0.05 ).While the relative levels of caspase-3,8,9 mRNA were 5.62 ± 0.65,5.41 ± 0.53,7.22 ± 0.51 in the experimental group and 1.00 ± 0.44,1.00 ± 0.21,1.00 ± 0.32 in control group at the proliferative phase.In the mean time,the relative levels of caspase-3,8,9 mRNA were 10.22 ± 0.72,25.3 ± 1.72,9.48 ± 1.89 in experimental group and 1.42 ± 0.14,1.14 ± 0.28,1.16 ± 0.12 in control group at the secretory phase,respectively (P ＜ 0.05).Compared with the control group,the levels of caspase protein in the experimental group were increased 2.04 and 1.60 folds in caspase-3,4.23 and 1.49 folds in caspase-8,2.65 and 3.5 folds in caspase-9 at the proliferative phase and at the secretory phase,respectively (P ＜ 0.05 ).ConclusionThe damaged model of ESC can be established after 48 hours by the withdrawal of 60 μmol/L mifepristone in treatment of ESC for 48 hours.

Aim To observe the effect of mineralocor-ticoid receptor blockade eplerenone on cell proliferation in obstructed kidney of rats. Methods Renal intersti-tial fibrotic animals were made with unilateral ureteral obstruction (UUO) and treated with eplerenone100 mg · kg - 1 · d - 1 . The kidneys were harvested on the 10th day and proliferating cell nuclear antigen ( PC-NA ), serum and glucocorticoid induced kinase-1 (SGK-1 ) and transforming growth factor-β1 ( TGF-β1 ) were detected with immunohistochemistry and Western blot. Results Renal histopathology showed large quantities extracellular matrix (ECM) accumula-tion in kidney with UUO, large numbers of inflammato-ry cells infiltrated in renal interstitium, renal tubular expansion and exfoliation of epithelial cells . The cell proliferation and ECM accumulation were inhibited in eplerenone treated rats significantly. Immunohisto-chemistry and Western blot showed that expressions of PCNA,SGK-1 and TGF-β1 were significantly up-regu-lated with UUO and down-regulated by eplerenone. Conclusion Eplerenone plays the role in inhibiting the cell proliferation and reducing ECM accumulation by down-regulating expression of SGK-1 pathway in rats with unilateral ureteral obstruction.

Aim To observe the effect of eplerenone(EPL) and Chinese decoction on cell apoptosis in obstructive nephropathy rats.Methods Sixty male Wistar rats were randomly divided into sham group, UUO group, EPL group and ZY group(n=15).Except sham group, the rats in the other groups were ligated with unilateral ureteral obstruction(UUO) for renal interstitial fibrosis model.The rats were treated with eplerenone at 100 mg·kg-1·d-1 added to diet in EPL group, and orally 13.7 g·kg-1·d-1 decoction of Chinese medicine in ZY group.The kidneys were harvested on 14th day, the number of renal cell apoptosis were detected by TUNEL, and serum aldosterone and 8-OhdG were detected with radioimmunoassay and ELISA.Caspase-12, caspase-9, Bax and Bcl-2 were examined by immunohistochemistry and Western blot.Results The levels of serum aldosterone, serum and urine 8-OhdG and the number of positive apoptotic cells increased significantly in UUO rats compared with Sham group.The overexpression of caspase-9, caspase-12 and Bax and down-regulated Bcl-2 were obvious in UUO group(P<0.01).The level of 8-OhdG, expression of caspase-9, caspase-12 and Bax were down-regulated, and Bcl-2 expression was up-regulated in eplerenone and Chinese decoction treated rats(P<0.01).Conclusion Eplerenone and Chinese decoction could inhibit cell apoptosis induced by oxidative damage after UUO via caspases and(or) Bax pathway.

A sort of absorbable Bondi of dura, whose main body is glue capsule, to compensate the deficiency of previous craniotomy, which easily causes delayed epidural hematoma. This device will help conglutinate dura to skull plate tightly, to stop bleeding and other purposes.
Absorbable Implants ; Adhesives ; Craniotomy ; Dura Mater ; Hemorrhage ; prevention & control ; Prosthesis Design

BACKGROUND:As a kind of undifferentiated precursor cells,the phenotypic differentiation of bone marrow mesenchymal stem cells (BMSCs) remains immaturity,thus it presents weak rejection following transplantation.However,the in vitro directional differentiation of BMSCs into neuronal cells is easy affected by various factors.OBJECTIVE:To observe the immunomodulatory effect and the potential of BMSCs differentiate into neuronal-like cells.DESIGN,TIME AND SETTING:A contrast observation was conducted at the Department of Cytology,Third People's Hospital of Wuxi from January 2008 to March 2009.MATERIALS:Bone marrow was harvested from chips of cancellous or ilium bone dudng hip joint surgery.METHODS:Firstly,the BMSCs were separated and cultured to establish mixed lymphocyte reaction (MLR) system.Secondly,2 samples of peripheral blood mononuclear cells (1×10~5/well) were added into 96-well plate,and then BMSCs treated by mitomycin were added according to different ratios (BMSCs/peripheral blood monouclear cells).At the end,the cells were cultured as follows:Method 1:DMEM+10% fetal calf serum+1 μmol/L RA +20 μg/L basic fibroblast growth factor+20 μg/L epidermal growth factor.Method 2:DMEM+2% dimethyl sulfoxide +100 μmol/L butylated hydroxyanisole.MAIN OUTCOME MEASURES:The growth rate of lymphocyte was measured by ~3H-Thymidine,and the effect of BMSCs on lymphocyte proliferation was observed.Additionally,the differentiation potential of BMSCs into neuronel cells was determined by immunofluorescenca and immunohistochemistrical staining.RESULTS:①The BMSCs inhibited lymphocyte proliferation in MLR system and the influence on proliferation of lymphocyte was direct related to ratio of BMSCs.②Under a light microscope,cytoplasm of BMScs were shrinkd,which presented typical perikaryon morphology at hour 2 after culture with method 1.The majority of BMSCs were formed neuronal-like cells without number changes at hours 3-5,which turned to be dipolar or multipolar neuronal shapes at day 3.There were 60%-70% neuronspecific enolase,45%-50% glial fibrillary acidic protein were positive expressed.However,the positive rate of nidogen was decreased 3.4%.Cells cultured with method 2 became smaller after 2 hours,formed dipolar or multipolar body cells,and most of cells were died after 48 hours.The 40%-50% neuronspecific enolase,35%-40% glial fibrillary acidic protein was found to be positive.The positive rate of nidogen was temporary increased to 63% at hour 2 after culture；however,it was decreased to 1.6% after 48 hours.CONCLUSION:BMSCs can differentiate into neuronal-like cells,as well as inhibit lymphocyte proliferation in MLR system,which possess down regulation effect on alloimmunity-reaction.

Objective This study observed the clinical curative effects of aldose reductase inhibitor epalrestat in treat-ment of diabetic sensory neuropathy. Methods Thirty-four diabetic sensory neuropathy patients were selected. The nerve electrophysiological data were collected before and after treating with epalrestat. Results The ratios of nerve con-ductive velocity in peroneal nerve sensory showed slowing down than normal before and after the treatment, which were respectively 61% and 32%; The ratios of the nerve conductive velocity in tibial nerve sensory nerve segment 1 showed slowing down before and after the treatment,which were respectively 97% and 65%; The nerve conductive velocity of the peroneal sensory nerve after treatment was significantly faster than that before treatment, the nerve conductive ve-locity of the the tibial nerve sensory nerve motion segments in 1 after treatment was significantly faster than that before treatment. Conclusion Epalrestat is one of the effective methods in the treatment of diabetic peripheral neuropathy.

Objective:To explore the safety key points of total pancreaticoduodenectomy in the era of vascular resection technology and the important factors affecting rapid postoperative recovery.Methods:The clinical data of 52 patients with pancreatic cancer who underwent total pancreaticoduodenectomy in Beijing Chaoyang Hospital Affiliated to Capital Medical University from November 2014 to September 2022 were retrospectively analyzed, including 34 males and 18 females, aged (62±9). The intraoperative situation, incidence of postoperative complication, postoperative blood glucose control and postoperative survival rate were analyzed.Results:All operations of the 52 patients were successfully completed, including 48 patients underwent total pancreaticoduodenectomy combined with portal vein resection and allograft vascular grafts via artery approach. The portal vein occlusion time was (20±5) min. The incidence of postoperative complications was 28.8% (15/52), including 2 cases of abnormal gastric empty, 2 cases of diarrhea, 2 cases of chylous fistula, 4 cases of abdominal infection, 1 case of gastrointestinal fistula, 3 cases of gastrointestinal bleeding, and 1 case of pulmonary infection. Subcutaneous short-acting insulin injection was used to control blood glucose in the early stage after surgery, short-acting insulin combined with long-acting insulin was used for subcutaneous injection before sleep for diet recovery. All patients did not experience uncontrolled hyperglycemia. The median survival time of 52 patients was 13 months, and the longest follow-up time was 38 months. There were 37 patients died of tumor recurrence, 4 patients died of cardiovascular and cerebrovascular accidents, and 1 patient died of pulmonary infection in the 42 died patients.Conclusions:Total pancreaticoduodenectomy via artery approach can improve the R 0 resection rate in pancreatic cancer patients with vascular invasion, the rate of postoperative complication and mortality has no significant increase. The postoperative blood sugar control is satisfactory and the quality of life is guaranteed.