OBJECTIVETo determine the interaction between miR-21 and DNA methylation in different breast cancer cells.

METHODSFluorescence tagged miR-21 inhibitor and its negative control (NC) were transient transfected into MCF-7 and MDA-MB-231 cell, the transfection efficiency was observed using fluorescence microscopy, and the miR-21 expression level and genome DNA methylation status before and after transfection were assessed by real-time PCR and bisulfite-qMSP respectively. To investigate the regulation effect of DNA methylation on miR-21, cells were treated with 5-AZA (2.5 µmol/L) for 72 h, with dimethyl sulfoxide (DMSO) treatment as its negative control (NC), and the expression level of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and AKT(also known as Protein Kinase B), two downstream genes of miR-21 were detected by Western blot.

RESULTSThe expression of miR-21 in MCF-7 cell was significantly knocked down (P < 0.01) by miR-21 inhibitor, with the genome DNA methylation level (P < 0.05) and all the three Dnmts: Dnmt1, Dnmt3a, and Dnmt3b unregulated. In contrast, the miR-21 expression in MDA-MB-231 cell was elevated ( P < 0.01) by miR-21 inhibitor, meanwhile, down- regulated of genome DNA methylation (P < 0.05) and Dnmt3b expression, upregulation of Dnmt3a were also observed. In addition, treated with 5-AZA resulted in significant increases of miR-21 expression in both MCF-7 and MDA-MB-231 cells (P < 0.01), with the protein level of PTEN increased in MCF-7 cell, which was further involved in the downregulation of AKT.

CONCLUSIONThe regulation effects of DNA methylation by transient transfection of miR-21 in MCF-7 and MDA-MB-231 cells are almost opposite, whilst the expression of miR-21 in two cell lines were all upregulated by decreased DNA methylation level and our results may provide some experimental evidences for the future development of rational therapy for different breast cancer.

Azacitidine ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MCF-7 Cells ; MicroRNAs ; genetics ; PTEN Phosphohydrolase ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins c-akt ; metabolism ; Real-Time Polymerase Chain Reaction ; Up-Regulation

Objective To explore the culturing strategies,curiculum provision,courses conferring methods,teaching effects as well as the associated managerial evaluations on the basis of Sino-French cooperation on medical education with the hope of summarizing helpful suggestions to Sino-Foreign cooperation on medical education. Methods The achievements of our Sino-French cooperation on medical education were analyzed and compared in the teaching models,culturing strategies along with courses conferring processes among seven-year medical students from both English-teaching and French-teaching classes. Results Our Sino-French cooperation on medical education was featured in its distinct culturing purposes and effective teaching model.Its scientifically formulated culturing strategy found its full expression in French-teaching atmosphere.The Sino-French cooperation on medical education was consistently welcomed and favorably recommended by both faculties and students. Conclusion The Sino-French cooperation on medical education has not only gained precious experience in culturing the cutting-edge medical talents with the international visions but also conduced to fulfill the goal to establish a modernized and internationalized medical school.

Objective In order to study the difference of depression-like behaviours among three widelyused stress models in rats.Methods The new-born Spragne-Dawley rats were randomly divided into maternal deprivation(MD) group(n=27),chronic mild stress(CMS) group (n=29),dual stress(DS) group ( n=31 ) and control (C) group ( n =30) on postnatal day 1.MD rats only received maternal deprivation.CMS rats only received chronic mild stress when 10 weeks old.DS rats received both maternal deprivation and chronic mild stress.Control rats received no experimental handling.Behavior tests including forced swimming test and sucrose consumption which were carried out to evaluate rats' depression-like behaviors in the thirteenth weeks.The extend time of floating and sucrose preference ratio should be recorded in the forced swimming test and sucrose consumption test respectively to reflect the behavior of helplessness and anhedonia of rats.Results In the forced swimming test,the extend floating time of MD group( 119.30 ± 65.56) s,CMS group ( 145.00 ± 80.24) s and DS group ( 170.03 ±61.75 )s were longer than the control group(81.14 ± 52.40)s (F =11.53,P ＜ 0.01 ).In the sucrose consumption test,the MD group(0.32 ± 0.22) had a low sucrose consumption.The comparison of the MD group and CMS group (0.43 ± 0.28 ) to the control group (0.54 ± 0.28 ) had significant differences (F =4.33,P ＜ 0.01 ).In these four groups,no sexual difference was found in the forced swimming test and sucrose consumption test ( all P ＞ 0.05 ).Conclusion The results suggest that MD,CMS and DS may induce some kind of depression-like behaviors in rodents such as anhedonia or the behavior of helplessness and the depression-like behaviors induced by different stresses are similar in male and female individuals.

AIM: To determine the optimal operation method and reduce operative complications by retrospective analysis of outcomes of different operative manipulation for patients with congenital ectopia lentis ( CEL) .
●METHODS: A retrospective study ranged from Jan. 1, 2010 to Jan. 1, 2015 was conducted and 31 CEL patients (57 eyes) treated with different operations were taken into this study. The postoperative outcome, operation related complications was analyzed in terms of different operative manipulations.
● RESULTS: The most common types of CEL are idiopathic and Marfan - related CEL. All eyes ( 98%) benefited from operation except 1 Marchesani syndrome eye concurrent with optic atrophy induced by glaucoma. A total of 3 eyes ( 5%) were treated by phacoemulsification combined with intraocular lens implantation; 8 eyes ( 14%) were treated by phacoemulsification, capsular tension ring implantation combined with intraocular lens implantation, among these 8 eyes, 1 eye was treated with capsular tension ring implantation combined with suture fixation; 39 eyes ( 68%) were treated by lens excision and anterior vitrectomy through corneal incision combined with intraocular lens implantation with suture fixation, among these 39 eyes, 2 eyes were treated with trabeculectomy , but not combined with intraocular lens implantation; 5 eyes ( 9%) were treated by intracapsular cataract extraction and anterior vitrectomy combined with intraocular lens implantation with suture fixation; 2 eyes ( 4%) were treated by lens excision and vitrectomy through pars plana combined with silicon oil injection.
● CONCLUSION: Almost all patients can obtain satisfactory outcomes through various operative manipulations. Phacoemulsification combined with intraocular lens implantation was the first choice for patients with lens dislocation range less than 90°. Phacoemulsification, capsular tension ring implantation combined with intraocular lens implantation was recommended for 90° - 180°, and lens excision and anterior vitrectomy through corneal incision combined with intraocular lens implantation with suture fixation was more suitable for lens dislocation range more than 180°. The appropriate operative methods for different types of CEL patients is the key to success.

Obejective To study the effect of daidzein(Daid) on the enzymes of bone metabolism and bone rebuilding in ovariectomized rats.Method Sixty 3-mon female SD rats were divided into 6 groups:Sham,OVX,OVX-D1～D4 groups.Every day the rats in Sham and OVX groups were given distilled water,and the rats in OVX-D1,OVX-D2,OVX-D3,OVX-D4 groups were given 25,50,75,100 mg/kg bw Daid respectively.After 6 mon,all of rats were killed,and the blood,liver and femur were collected.The content of calcium and phosphorus in serum and the enzymes of bone metabolism were measured.Results The content of calcium,phosphorus and ATPase in serum was increased,but the activity of serum alkaline phosphatase(ALP) and tartrate-resistant acid phosphatase(TRAP-5b) was decreased dose-dependently in OVX-D groups compared with OVX group.Conclusion The expression of bone metabolism-associated enzymes could be inhibited,which resulted in inhibition of bone formation and absorbsion in ovariectomized rats.The new balance between bone formation and absorption could be rebuilt by Daid,through the mechanism of decreasing rate of bone conversion.

OBJECTIVE:To study pharmacokinetics of domestic and imported Phenazopyridine hydrochloride tables and bioavailability of domestic tablets, and to evaluate the bioequivalence of two kinds of tablets. METHODS: A randomized crossover design was performed in 18 healthy male volunteers. They received a single oral dose of domestic or imported tablets 200 mg. Plasma concentration of phenazopyridine hydrochloride was measured by HPLC. The pharmacokinetic parameters were calculated by 3p97 software and relative bioavailability was evaluated. RESULTS: The plasma concentration-time curves of domestic and imported tablets conformed to one-compartment model. Main pharmacokinetic parameters of domestic tablets vs. imported tablets were as follows: t1/2Ke(3.52?2.03) h vs. (3.18?1.85)h; tmax(0.76?0.33) h vs. (0.79?0.43)h; Cmax(76.41?70.15) ng?mL-1 vs. (75.49?70.37) ng?mL-1; AUC0～8(159.10?116.32) ng?h?mL-1 vs. (164.65?129.89) ng?h?mL-1; AUC0～∞(237.12?115.06) ng?h?mL-1 vs. (262.69?155.05) ng?h?mL-1. The relative bioavailability of domestic tablets was (96.63?14.05)% compared with imported tablet. There was no significant difference between the pharmacokinetic parameters of two formulations by variance analysis, t-test and 1-2? confidence interval method. CONCLUSION: The domestic and imported tablets are bioequivalent.

Objective To detect left atrial (LA) function and left ventricular (LV)-LA coupling using vector flow mapping (VFM) and two-dimensional tissue tracking (2DTT) echocardiography in patients with diabetes.Methods A total of 51 patients with type 2 diabetes (DM group) and 38 healthy volunteers (control group) were studied.LA and LV strain were assessed by 2DTT.The energy loss (EL) of LA and LV during ventricular systole (ELs),early diastole (ELed),and atrial contraction (ELac) were measured by VFM.Results LVEL in DM group was significantly increased compared to that in control group (P<0.05).LAELs and LAELed in control group were higher than those in DM group (P<0.05);while LAELac decreased in control group (P<0.05).Multivariate regression analysis identified E and LVELed as independent predictors of LAELed;Peak torsion,LA peak systolic strain and LVELac were independent predictors of LAELac.Conclusions The reservoir and conduit function of LA are impaired in patients with DM,while the pump function increases as a compensation.Abnormal LA-LV coupling also appears in patients with DM.

Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

Objective To investigate the value of Cytochrome P450 (CYP3A5) * 3 gene polymorphism in providing individualized administration for the use of tacrolimus (Tac) in renal transplantation recipients.Method Pyrophosphate sequencing method was used to determine the CYP3A5 * 3 genotype of renal transplant patients in the first day after surgery.Sixty recipients were divided into experiment group and control group.Both groups of patients were routinely given the initial dose of Tac-4.0 mg/day in the first day after surgery.The experiment group of patients were given different doses of Tac based on the different CYP3A5 * 3 genotypes at the third day after surgery [for AA:0.12 mg/(kg· day),and for GG:0.06 mg/(kg· day)],and the control group of patients were given different dosages of Tac according to drug concentration.Different parameters were compared between two groups of patients:percentage of patients reaching the target concentration (3-8 μg/L) at the fifth day after surgery,days required to reach the target concentration level,times needed to adjust the dosage of Tac within two weeks.Result The percentage of patients reaching the target concentration in experiment group and control group was 90％ and 46.67％,respectively (P＜ 0.05).Days required to reach the target concentration were (3.67 ± 1.32) and (7.57 ± 3.42) on average,respectively (P ＜ 0.05).Times of adjusting the Tac dose in experiment group was significantly less than those in the control group (P＜0.05).In the experiment group,the target concentration was obtained even without dosage adjustment (70％).Conclusion Individualized adjustment of Tac doses for patients according to recipients' different CYP3A5 * 3 genotypes is beneficial for reaching target concentration as soon as possible,which is superior to traditional dosage regimen.

Objective To explore life experience in chronic obstructive pulmonary disease (COPD) patients with the habit of exercise for more than 5 years,to enhance the confidence of participating in the rehabilitation in other patients, to help the medical staff pay more attention to guide the patients with COPD to take up the rehabilitation exercise. Methods The narrative research method in qualitative research was used to analyze the collected data. Results The study found that the coping approach of the contradiction between exercise and dyspnea, the ability to live together with the disease peacefully and the enthusiasm of persuing life were deserved to be learned. Conclusions Full of perseverance and persisted rehabilitation exercise may alleviate the symptoms, reduce the number of acute exacerbation of COPD and improve the quality of life for COPD patients. Medical staff should encourage COPD patients to participate in the rehabilitation in the right time with appropriate methods.