Objective:To investigate the influential factors on the formation of mammospheres from MCF-7,MDA-MB-231 and primary breast cancer epithelial cells,and to detect the proportion of CD44+/CD24-/low among them.Methods:MCF-7,MDA-MB-231 and primary breast cancer epithelial cells were cultured in suspension to generate primary mammospheres.After 7 days of cultivation,the proportion of cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry.Results:MCF-7 had the highest efficiency of mammosphere formation(2.1%?0.3%),while MDA-MB-231 cells hardly had any mammosphere formation,and no mammosphere formation was observed in breast cancer epithelial cells.MCF-7 tended to adhere under stem cell culture conditions without B27.Flow cytometry analysis indicated that,compared with 2.0%?0.1% for the MCF-7 monolayer culture cells,11.8%?0.3% and 8.2%?0.8% of the MCF-7 mammosphere cells from MCF-7 under different conditions were CD44+/CD24-/low(P

Objective To study the inhibit efficiency of lake sediment phosphorus release by adding chemical reagents and to provide the technical basis for the comprehensive management of city lake.Methods In May 20,2008,Mochou Lake upper sediment and water samples were collected.The phosphorus forms of sediments,total phosphorus(TP),NaOH-P,HCl-P,inorganic phosphorus(IP),organic phosphorus(OP) were analyzed and the pH value and content of TP in water samples were determined.Each water(250 ml)-sediment(10 g) system was respectively added by 0-1 000.0 mg/L of Ca(OH)2,2.0-10.0 mg/L of A12(S04)3 and 0-25.0 mg/L of FeCl3.The pH value and content of TP in water were determined,then the control rate of TP release were calculated.Results The pH values were between 7.6-8.0,TP content was 2.0 mg/L in water of Mochou Lake.The contents of TP,HCl-P,NaOH-P,IP,OP were 2 187.06,1 383.89,526.48,1 910.37 and 276.69 mg/kg respectively.With the increase of Ca(OH)2 concentration,pH value of water increased rapidly,content of TP showed a decreasing trend,control rate of TP release was increased.With the increase of A12(SO4)3 or FeCl3 concentration,pH value of water declined slightly,content of TP showed a decreasing trend,control rate of TP release was increased.Conclusion Ca(OH)2,A12(SO4)3,FeCl3 can significantly inhibit the phosphorus release of sediments in Mochou Lake.

Objective To discuss magnetic resonance imaging(MRI)scanning technology for ankle joint and its clinical value.Methods 42 cases of ankle joint trauma were examined with MRI using GPFLX coil.It was performed through sagittal T2WI,T2WI F/S,T1WI;coronal T2WI;axial T2WI.FOV was big enough for the whole trauma ankle joint.To make image clear,some soft materials should be placed between the trauma ankle joint and the GPFLX coil.The injured ankle joint was not allowed to move.The technology of No Phase Wrap is also necessary.Results The high quality rate was up to 90% for the ankle joint.Among 42 cases,6 had traumatic arthritis,14 had ankle joint contusion,9 had Achilles tendon rupture,6 had other structural injuries,4 had late degenerative change,and 3 had normal images.Conclusion MRI is a sensitive technique for injured ankle joint.It can accurately show the degree and location of ankle joint trauma.It has an important clinical significance in finding bone contusion and other structural injuries,and it is very helpful for treatment.

Objective To explore the inhibition effects of epidermal growth factor receptor (EGFR) antagonist monoclonal antibody cetuximab (C225) on breast cancer stem cells in human breast cancer cell line MCF-7.Methods The effects of C225 on the proliferation of breast cancer cell line MCF-7 were detected by MTT assay. MCF-7 cells were cultured to generate primary mammospheres, and were divided into control group, C225 group, epidermal growth factor (EGF) group and EGF + C225 group according to whether or not the culture media contained exogenous EGF and C225.Thirteen days after culture, the volume and number of mammospheres of these four groups were observed, and mammosphere-forming efficiency ( MFE) was calculated. The percentages of CD44~+ CD24~- cells in mammospheres of these four groups and in routinely cultured MCF-7 cells were determined by flow cytometry. Results The inhibition rate on MCF-7 cells increased with the concentration of C225.Compared with control group, the volume of mammospheres in C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [(0.61 ±0.04)% vs (1.44±0.09)%, P<0.01; (3.50±0.29)% vs (9.07 ±0.52) % , P<0.01]. Compared with EGF group, the volume of mammospheres in EGF + C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [ (0.68 ± 0.04) % vs (1.61 ± 0.05) % , P < 0.01; (4.00 ± 0.58) % vs (10.47 ± 0.79) % , P < 0.01]. The percentage of CD44~+ CD24~- cells in routinely cultured MCF-7 cells was (2.03 ±0.15) % , and was significantly different from those in EGF group and control group (P < 0.01). There was no significant difference in volume of mammospheres, MFE and percentage of CD44~+ CD24~- cells in mammospheres between EGF group and control group (P >0.05). Conclusion C225 has significant inhibition effects on CD44~+ CD24~- cells in MCF cells.

OBJECTIVE:To investigate the effects and mechanism of total flavonoids from Armeniaca mume on depression in chronic stress depression model rats. METHODS:60 rats were randomly divided into normal saline group,model group,fluox-etine group(positive control,20 mg/kg)and total flavonoids from A. mume low-dose,medium-dose,high-dose groups(80,160, 240 mg/kg), 10 in each group. Except for normal saline group, the other groups adopted chronic unpredictable mild stress (CUMS)+solitary feeding condition to induce depression model. These groups were intragastrically administered,once a day,for 28 d. Changes of body mass and food intake,degree of preference for sugar were observed;forced swimming test and tail suspen-sion test were used to determine the time that rats did not move;open field test was used to determine the changes of residence time in central square,horizontal crossing lattice,standing times,modification times;tumor necrosis factor α(TNF-α),cortisol, interleukin 6 (IL-6),serotonin (5-HT) levels in serum after 24 h of last administration were determined. RESULTS:Compared with normal saline group,growth of body mass,food intake and sugar preference percentage in model group were decreased;the time that rats did not move was prolonged in forced swimming test and tail suspension test;residence time in central square was prolonged,while horizontal crossing lattice,standing times and modification times were decreased in open field test;serum levels of TNF-α,cortisol were increased,while IL-6,5-HT were decreased,with statistical significances (P<0.05 or P<0.01). Com-pared with model group,except that there were no obvious improvement in sugar preference percentage,the time that rats did not move in forced swimming test,modification times in open field test in total flavonoids from A. mume low-dose group and IL-6, 5-HT levels in serum in total flavonoids from A. mume low-dose,medium-dose groups,the above-mentioned indexes were obvious-ly improved in other groups (P<0.05 or P<0.01). CONCLUSIONS:Total flavonoids from A. mume can obviously improve the CUMS-induced depression in rats,and the mechanism may be related to inhibiting inflammation response,adjusting the hypothala-mus-pituitary-adrenal axis functions.

Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.

Objective To evaluate effect of intensive insulin treatment(IIT)on healthcare-associated infection (HAI)rate in patients with acute stroke and stress hyperglycemia.Methods Databases,including PubMed,Em-base,Cochrane Library,WanFang,and China National Knowledge Infrastructure(CNKI)Data,were electronically searched,relevant journals and references of the included literatures were also searched manually,literatures were selected according to the uniform inclusion and exclusion criteria,incidence of HAI and mean blood glucose in patients who received IIT for acute stroke were assessed systematically.Results A total of 13 randomized controlled trials (RCT)involving 1 032 patients were included in this systematic review.Meta-analysis results showed that 10 studies involving 832 patients were finally enrolled for comparing HAI rate, HAI rates in IIT group and conventional insulin treatment group were 28.3% and 56.1 %,respectively(Z =4.50 ),difference between two groups was statistically significant (RR=0.53 [95 %CI :0.40 to 0.70],P <0.001 );A total of 328 patients in 5 studies were finally included in the comparison of blood glucose,difference in mean blood glucose between two groups was statistically significant(MD =-2.52 [95% CI :-4.30 to -0.74],P =0.006).Funnel plot of HAI rate revealed that there was publication bias.Conclusion IIT is used for the regulation of stress hyperglycemia in acute stroke,it can reduce the incidence of HAI and blood glucose in patients.

Objective To summarize the perioperative nursing of huge malignant lobulated breast tumor, so as to promote the recovery of patients. Methods A total of 11 patients with huge malignant lobulated breast tumor patients were selected, assessed the preoperative status,finished breast tumor site nursing, strengthen psychological nutritional assessment, the adjustment of the respiratory system for the operation, prevent falls. In addition to do a good job in general nursing and postoperative pain management, but should also focus on the observation of the chest wall flap, drainage tube care, to prevent bleeding and infection, etc. Results By implementing the nursing plan, 1 case of breathing difficulties preoperative patients with respiratory function improved, 1 case of refuse treatment of patients accepted surgical treatment after psychological intervention, 2 cases of postoperative patients with skin flap necrosis, local treatment, after compression bandage, 1 case of good recovery, 1 case of poor treatment effect for the wound recovered well after back surgery graft;1 case of patients with subcutaneous effusion, to adjust the drainage tube, suction fluid, after compression bandage, partial treatment, recovering well. Patients were good recovery, discharged smoothly. Conclusions By constantly sum up experience, through positive assessment of patients after preoperative mental and physical health and attaches importance to patients' skin flap, bleeding, pain, completes the drainage tube nursing and prevention of infection, can effectively promote the recovery of patients.

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.