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

To improve the yield of industrial fermentation, a method based on near infrared spectroscopy was presented to predict the growth of yeast.The spectral data of fermentation sample were measured by Fourier-transform near-infrared (FT-NIR) spectrometer in the process of yeast culture.Each spectrum was acquired over the range of 10000-4000 cm1.Meanwhile, the optical density (OD) of fermentation sample was determined with photoelectric turbidity method.After that, a method based on competitive adaptive reweighted sampling (CARS) was used to select characteristic wavelength variables of NIR data, and then extreme learning machine (ELM) algorithm was employed to develop the categorization model about the four growth processes of yeast.Experimental result showed that, only 30 characteristic wavelength variables of NIR data were selected by CRAS algorithms, and the prediction accuracies of training set and test set of the CARS-ELM model were 98.68% and 97.37%, respectively.The research showed that the near infrared spectrum analysis technology was feasible to predict the growth process of yeast.

Acupuncture Therapy ; Adult ; Chronic Disease ; therapy ; Humans ; Male ; Moxibustion ; Pelvic Pain ; therapy ; Prostatitis ; therapy ; Treatment Outcome ; Young Adult

Bicuspid ; Dental Pulp Cavity ; pathology ; Humans ; Mandible ; Root Canal Therapy

In this experiment six methods,calcium chloride(CaCl_(2)) precipitation,polyethylene glycol(PEG,pH7.0) precipitation,polyethylene glycol(PEG,pH11.5) precipitation,aluminum chloride(AlCl_(3)) precipitation,Amicon Utcra centrifugal filter devices and cellulose nitrate membrane were used to concentrate the vaccine poliovirus type 1(PV_(1)) added to water body;experimental conditions for concentration were selected and optimized.The results showed that two methods,CaCl_(2)and PEG(pH 7.0) precipitation were suitable for concentrating virus in large volumes of water while amicon utcra centrifugal filter devices for small ones.The virus recovery of the three methods reached a 100% rate.

Objective To explore clinical value of the color Doppler ultrasonic(CDU)and TNM staging of renal cell carcinoma.Methods 265 cases of renal cell carcinoma(RCC)confirmed by surgical pathology were retro-spectively analyzed.According to the WHO 2004 pathological types of classification standards and the American Joint Committee on Cancer(AJCC)cancer staging manual/6th TNMstaging criteria.The analyzed application of a CDFI to TNMstaging of renal cell carcinoma,and surgical pathology results werecompared.Results 265 cases of RCC and right kidney in 136 cases,129 cases of left kidney;Diameter of 1.5 -12.8cm;Pathological diagnosis of renal clear cell carcinoma in 226 cases(85.28%),28 cases of papillary renal cell carcinoma(10.57%),renal too color cell car-cinoma in 5 cases(1.89%),4 cases of multilocular cystic renal cell carcinoma(1.51%),Bellini duct carcinoma in 2 cases(0.75%).Diagnosis results of CDU and pathological in 250 cases(94.34%),CDU misdiagnosis leakage 15 cases(5.66%).Within the kidney showed low echo or high echo,and mixed echo lumps,and invasive growth to the surrounding tissues or organs sign for it.CDU showed the various blood flow signals were given priority to with renal clear cell carcinoma;CDU and surgical pathology TNMstaging coincidence rate was 90.40%(226 /250),inclu-ding T1 N0 -1 M0 ,T2 N0 -1 M0 ,T3 N0 -2 M0 -1 and T4 N0 -2 M0 -1 of coincidence rate were 92.12%(152 /165),85.71%(36 /42),82.14%(23 /28)and 100.00%(15 /15).Conclusion CDU can be used as the preferred method of diag-nosis and evaluation of renal cell carcinoma,which can can provide valuable information for TNMstaging.

Objective To compare the effects of subgluteal(SG) and sub-subgluteal-fold(SSGF)approach for ultrasound-guided siatic nerve block. Methods One hundred forty-eight patients undergoing lower limb surgery were randomly divided into two groups to receive SG approaches and SSGF approaches to sciatic nerve block under real time ultrasound guidance. A combined posterior lumbar plexus block under ultrasound guidance was performed for sufficient surgery anesthesia. 20 ml of 0. 5% ropivacaine was used for sciatic nerve and lumbar plexus block separately. Measurements included skin-to-nerve distance,reorientation of the needle during block and execution time,rates of sensory and motor blockade after 15 min and 30 min of injection, quality of surgery blockade, duration of the sensory and motor block, and postoperative complications related to sciatic nerve block. Results In SSGF group, execution time and reorientation of needle for sciatic nerve block was significantly less than those of the SG group( P ＜0.01).But motor blockade in the SG group was quicker when compared with SSGF group ( P ＜0.01). There were no significant differences in the quality and duration of blockade between the two groups. Conclusions Both SG and SSGF approach can be used for sciatic nerve block with equal sensory and motor block rate,whereas sciatic nerve block via SSGF approach was faster and easy to perform than the SG one.

n positive blood culture, and they are resistant to a variety of antimicrobial agents, which should be called attention.

Objective To investigate the pathogen distribution and susceptibility profile of fungal isolates from bloodstream infections,and valuate the clinical utility of G test in diagnosis of fungal infections for the purpose to improve antifungal therapy.Methods A retrospective analysis was carried out to analyze the fungal pathogens isolated from bloodstream infections in the First Affiliated Hospital of Nanjing Medical University during the period from January 2013 through December 2015 and their antimicrobial susceptibility.Results A total of 114 fungal strains were isolated from bloodstream infections during the 3-year period,most of which were Candida (99/114,86.8％),especially Candida albicans (30.7％).About 41.2％ (47/114) of the fungal strains were isolated from Department of Thoracic Surgery (10,5 and 4 strains in 2013,2014 and 2015),Hematology (11 strains in 2014),and ICU (7 strains in 2014).Antimicrobial susceptibility testing showed that all the fungal strains (100％) were susceptible to amphotericin B,but 83.5％ susceptible to itraconazole (the lowest).G test was positive before the result of blood culture in 13 of the 54 patients who received G test.Conclusions Candida was the most common fungus in fungal bloodstream infection.Amphotericin B is the most active antifungal agent in vitro.Blood culture combined with serological test can provide clinicians an earlier and reliable diagnosis.

Objective:To explore clinical application value of color Doppler ultrasound (CDU) assigned to diagnose triple-negative breast cancer (TNBC) and axillary lymph node metastasis (ALNM).Methods:A retrospectively analyzed of proved of clinical data on the 863 patients breast cancer in women by CDU inspection,operation and pathology.According results will be divided into TNBC group and non negative breast cancer (NTNBC) group via clinical pathological and immunohistochemical detection.Using multi-factor unconditioned Logistic regression analysis the CDU supposed the diagnosis and the correlation of TNBC.Results:The coincidence rate of CDU and pathological diagnosis was 90.96％ (785/863),78 cases (9.04％) remove breast masses,105 patients TNBC group and NTNBC group 680 patients.The mass circular or elliptic (48.57％ vsl0.70 ％),the tumor maximum diameter 5.0 cm (21.90％ vs 15.30 ％),micro calcifications (87.62％ vs 74.12 ％),state clearly (56.19％ vs 41.47 ％),no burr edge character (74.29％ vs 41.62 ％),peripheral hyperechoic halo ring (74.29％ vs46.76 ％),rear echo attenuation (72.38％ vs 46.76 ％),no ALNM (61.90％ vs 37.35％) comparing the two groups exists significant difference (P＜ 0.05),while the mass number,and vertical and horizontal diameter ratio,degree of echo,echo distribution and blood flow classification comparing the two groups were no significant difference (P＞ 0.05).The CDU fu points ＞ 10 divided into NTNBC and pathological coincidence rate 93.93％ (680/724),8 ～ 10 points TNBC suspected diagnosis coincidence rate 11.51％ (16/139),＜ 8 points TNBC tendency diagnosis coincidence rate 64.03％ (89/139),TNBC total diagnostic coincidence rate was 75.54％ (105/139).In CDU diagnosed 785 cases of breast cancer,the ALN enlargement in 319 cases (40.64％),with TNBC group was 61.90％ (65/105),NTNBC group was 37.35％ (254/680).Logistic regression analysis results:tumor assumes the circular or elliptic,edge without burr,peripheral hyperechoic halo ring,rear echo attenuation and ALNM with TNBC has certain correlation (P＜0.05).Conclusion:The seemingly benign breast diseases and axillary lymph node enlargement,the CDU,supposed less than 8 points,age 50 or less or not at the same time,menopause or having a history of breast cancer susceptibility to family,important reference index,clinical value has high of CDU diagnostic of TNBC and ALNM.