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.

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

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

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 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.

Objective To determine the levels of soluble intercellular adhesion molecule-1(SICAM-1) in serum and induced sputum in children with mycoplasma pneumoniae pneumonia(MPP),and to investigate the role of SICAM-1 in pathogenesis of MPP.Methods The levels of SICAM-1 in serum and induced sputum and sputum cell count were measured by enzyme linked immunosorbent assay(ELISA) kit in 44/34 children with MPP in acute episode period,36/28 children in remission episode period and 38/30 healthy children.Results The levels of serum SICAM-1,sputum SICAM-1 and percent of neutrophil(N),lymphocyte(L),eosinophile(EO),mast cell(Ma) were significantly higher in acute episode period in MPP than those in remission episode period and control group,respectively((all P0.05);)the indexs in remission period and in controls showed no marked difference.The le-(vels of) serum and sputum(SICAM-1) in children with asthma attack in acute and remission episode periods in MPP increased significantly compared with those in children without asthma in both periods(P

AIM: To investigate the time - effect relationship between the expression of rhodopsin and recoverin and photoreceptor damage induced by N - nethl - N -nitrosourea ( MNU) .
METHODS: Thirty-six 7-week old Sprague-Dawley ( SD ) rats were intraperitoneally injected with MNU ( 60mg/kg ) and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection ( 6 per group) , respectively. As a control, six rats were injected with phosphate buffer saline (PBS) 5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling ( TUNEL ) and transmission electron microscope ( TEM ) in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.
RESULTS:The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak ( 29. 7% ±2.3%) at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection (P<0. 05), and extremely significant difference comparing to control group after 12h ( P<0. 01 ); while recoverin dramatic declined at 12h, and extremely significant difference after 24h (P<0. 01).
CONCLUSION:60mg/kg MNU intraperitoneally injection one - time may specifically induce photoreceptor apoptosis, The mechanism of down - regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.

Objective To analyze the distribution and subcellular localization of NOR1 alternative splice isoforms in human tissues and cell lines. Methods NOR1 open reading frame(ORF) was amplified from human fetal brain cDNA library and subcloned into pCMV/myc vector. The level of NOR1 mRNA in human tissues was determined by real-time RT-PCR. Region spanning exon 2 was amplified from cDNA or genomic DNA by specific primers and sequenced. The expression plasmids of NOR1 were transfected into cells and immunofluoresence assay was performed to determine the subcellular localization of NOR1 protein isoforms in human cells. Results The expression of NOR1 mRNA was high in human adult testis, moderate in human fetus nasopharynx, trachea, brain, and kidney tissues, and weak or undetectable in other tissues. Two splice variants of human NOR1 gene resulted from alternative splicing at exon 2 were identified. Both of isoform 1 and isoform 2 mRNA were detected in human fetus brain. Isoform 2 was the sole isoform in other tissues but brain. Only isoform 2 mRNA was detected in cell lines used in this study, though no genomic deletion of exon 2 could be found in all these cell lines. Immunofluoresence assay showed both isoform 1 and isoform 2 proteins were distributed in cytoplasm. Conclusion Alternative splice isoforms of tumor suppressor gene NOR1 are identified. NOR1 isoform 1 and isoform 2 are both detected in fetus brain. NOR1 isoform 2 lacking of exon 2 is the sole isoform in multiple tissues except for brain. The exon 2 encoded peptide does not affect the subcellular location of NOR1 protein.