Objective To evaluate the effects of metformin on transforming growth factor-betal (TGF-β 1)-induced epithelial-mesenchymal transition (EMT) in and invasion of the human melanoma cell line 1205Lu.Methods In vitro cultured 1205Lu cells were divided into 3 groups to be treated with serumfree culture medium (blank control group),serum-free culture medium containing 5 ng/ml TGF-β 1 (TGF-β 1 group) and serum-free culture medium containing 5 ng/ml TGF-β 1 and 1 mmol/L metformin (TGF-β1 + metformin group),respectively.After 48-hour treatment,morphological changes of 1205Lu cells in the above groups were observed by using a microscope,and photos were taken at the same time.Transwell invasion assay was performed to estimate cellular invasive activity,Western blot analysis and real-time fluorescence-based quantitative RT-PCR were conducted to determine the mRNA and protein expression of N-cadherin and matrix metalloproteinase-9 (MMP-9),respectively.Statistical analysis was carried out by one-way analysis of variance (ANOVA) and least significant difference (LSD) test.Results Compared with the blank control group,the stimulation of TGF-β 1 could induce the epithelial-mesenchymal transition (EMT)-like morphological changes in 1205Lu cells,while TGF-β 1 combined with metformin could reverse the EMT-like morpho-logical changes.The number of 1205Lu cells crossing the transwell Matrigel per high-power field (× 200) was significantly higher in the TGF-β1 group (412.2 ± 13.427) than in the blank control group (194.1 ± 8.295) and TGF-β1 + metformin group (175.3 ± 8.693).Compared with the blank control group and TGF-β1 + metformin group,the TGF-β1 group also showed significantly increased mRNA and protein expression of N-cadherin (mRNA:6.678 ± 0.043 vs.1.000 ± 0.000,1.035 ± 0.015;protein:1.963 ± 0.016 vs.0.603 ± 0.029,0.207 ± 0.009) and MMP-9 (mRNA:5.351 ± 0.604 vs.1.000 ± 0.000,0.930 ±0.130;protein:1.243 ± 0.027 vs.0.575 ± 0.009,0.629 ± 0.008).Conclusion Metformin can obviously inhibit TGF-β1-induced EMT-like morphological changes in,the capacity to penetrate Matrigel-coated transwell chambers of and the mRNA and protein expression of N-cadherin and MMP-9 in 1205Lu cells.

Congresses as Topic ; Humans ; Multiple Myeloma ; United States

OBJECTIVE To promote the management of nosocomial infections continuous improvement in the clinical and medical technology department.METHODS According to the regulation of nosocomial infections management,a handbook of nosocomial infections management for clinical and medical technology department was designed,and the monitors of nosocomial infection could perform real time inspection and record according require of the handbook.The department of nosocomial infections management examined the monitoring work of clinical and medical technology department every month and summarized every year,and the results were internalized to the valuation of medical quality management.RESULTS After 3 years of the usage of the handbook,the capability of the monitor groups of nosocomial infection and the quality of the all monitoring items were significantly improved;the qualified rates were all above 96.30%.CONCLUSIONS The handbook of nosocomial infections management is useful to improve the quality of nosocomial infections management in the clinical and medical technology department.

Aim To observe the effects of pituitrin ad- ministrated intravenously on electrocardiogram (ECG), and the effects of taurine on the ECG changes induced by pituitrin in conscious unrestrained rats. Methods ECG(leadⅡ)was recorded telemetrically and analyzed with Data Science International (DSI, USA); the changes of the ECG T wave and the heart rate at different times after the intravenous(iv) injection of pituitrin with or without intraperitoneal (ip) pre-administration of taurine were observed and calculated. Results ① Pituitrin 0.25～4.0 U?kg -1 dose-dependently elevated the ECG T wave; the T wave values were increased 0.75～3.42 times over the values before pituitrin, and the maximal effect appeared at 15 s after the injection. ② TAU 100, 200 or 400 mg?kg -1,injected intraperitoneally 30 min before the pituitrin, inhibited the T wave elevation and heart rate decrease induced by intravenous injection of pituitrin; compared with NS+pituitrin group, T wave elevation(mV) was decreased from 1.77?0.165 to 1.60?0.13(P

Objective To construct recombinant plasmids containing various functional domains of APC protein and detect their expression in HCT-116 cells. Methods Five APC gene fragments were amplified by PCR with whole APC gene as template and primers designed according to APC cDNA sequence and mutation cluster domain. The five obtained fragments were cloned into eukaryotic expression vector pEGFP-N3 to generate recombinant pEGFP-N3-APC1-5. Sequence of the inserted gene was identified and analyzed after restriction enzyme digestion. Liposome-mediated recombinant plasmid pEGFP-N3-APC was transfected into HCT 116 cells and identified by green fluorescence. RT-PCR was employed to validate the expression of recombinant vectors in cells. Results Recombinant pEGFP-N3-APC1-5 were confirmed by restriction enzyme digestion and sequence analysis. The plasmids could be expressed in HCT-116 cell line detected by fluorescence microscope. Results of RT-PCR made clear that vectors constructed could be expressed in HCT-116 cells. Conclusion The relative efficient expression of five recombinant expressive vector in HCT-116 cell line may provide an experimental basis for selecting specific therapy peptide for colorectal cancer.

OBJECTIVETo research the clinical application of lower cervical pedicle screw fixation procedure.

METHODSFrom September 2011 to July 2013,32 patients underwent posterior pedicle screw-rod system fixation were retrospective analyzed includinig 20 males and 12 females with an average age of 56.4 years old ranging from 21 to 78 years. Among them, 10 patients were traumatic cervical spinal injury, 9 patients were cervical spinal canal tumors, 7 cases were posterior longitudinal ligament ossification of cervical vertebrae, 6 cases were multiple segmental cervical spondylopathy. Preoperatively, X-ray, computed tomography, magnetic resonance imaging and magnetic resonance angiography of the vertebral artery were performed in all patients. After the operation and during the follow-up,X-ray and computed tomography were performed to confirm the pedicle screw position. The accuracy of the pedicle screw placement was evaluated by 4 grades classification from Lee. The spinal cord function was assessed by ASIA impairment scale for traumatic patients and JOA score for non traumatic patients.

RESULTSTotally 144 pedicle screws performed on 32 patients from C3 to C7 involving 132 screws of grade 0,5 screws of grade 1,5 of screws grade 2 and 2 screws of grade 3 according to postoperative CT. There were 12 screws penetrating the pedicle cortex including 8 screws at lateral,2 screws at caudal, 1 screw at medial and 1 screw at cranial. The follow-up time was 12 to 33 months with an average of (21.0±1.5) months. The spinal cord function was not improved in 6 complete cervical spinal cord injury patients,but their paraplegic level descended 1 to 3 segments. Four incomplete cervical spinal cord injury patients' ASIA impairment scale was increased by 1 to 2 grades in average. The JOA score of 22 atraumatic patients increased from preoperative 11.5±0.8 to 15.9±0.6 of postoperative at 6 months (P<0.01). There were no screw loosening,screw pullout and screw-rod breakage.

CONCLUSIONThe lower cervical pedicle screw fixation can provide excellent 3D stability of the vertebral column. The operation risk and Complication could be minimized by adequate preoperative evaluation for appropriate cases and individual pedicle screw placement. It deserved the clinical expansion.

Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Female ; Humans ; Male ; Middle Aged ; Pedicle Screws ; Retrospective Studies ; Spinal Cord Injuries ; physiopathology ; surgery

Objective To explore the consistency of time-of-flight (TOF) technology of PET/MRI and PET/CT for max standardized uptake value (SUVmax) of body malignant tumors.Methods A retrospective analysis of TOF-PET/CT and TOF-PET/MR imaging data about twenty patients with body malignant tumors was performed.Patients were divided into two groups (each n=10),including PET/CT first and sequentially PET/MR group and PET/MR first and sequentially PET/CT group.Bland-Altman figure was used to evaluate consistency of SUVmax of malignant lesions between TOF-PET/CT and TOF-PET/MR.Multi-way ANOVA was used to analysis effect of machine type and exam order on SUVmaxof malignant lesions in TOF-PET/CT and TOF-PET/MR.Results SUVmax of malignant lesions in TOF-PET/CT and TOF-PET/MR had good consistency in two groups (PET/CT first and sequentially PET/MR group:Mean difference was 3.06,95％CI was [-7.5,13.6];PET/MR first and sequentially PET/CT group:Mean difference was 3.0,95％CI was [-2.4,8.3]).SUVmax was not influenced by machine type (F=0.005,P=0.95),but exam order (F=46.00,P＜0.001).Conclusion PET/MR and PET/CT with TOF technology have comparative diagnostic value in SUVmaxof body malignant lesions.SUVmax of body malignant lesions increases in delay time,which is not related to machine type,but exam time.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.

Twenty-three histone methyltransferase genes were obtained from transcriptome dataset of Lonicera japonica. The nucleotide and proteins characteristics, subcellular localization, senior structural domains and conservative forecasting were analyzed. The result of phylogenetic tree showed that 23 histone methyltransferases were mainly divided into two groups: lysine methyltransferase and arginine methyltransferases. The result of gene expression showed that 23 histone methyltransferases showed preference in terms of interspecies and organs. They were more expressed in buds of L. japonica than in L. japonica var. chinensis and lower in leaves of L. japonica than in L. japonica var. chinensis. Eight genes were specific expressed in flower. These results provided basis for further understanding the function of histone methyltransferase and epigenetic regulation of active ingredients of L. japonica.
Computational Biology ; Gene Expression ; Histone-Lysine N-Methyltransferase ; genetics ; Lonicera ; enzymology ; genetics ; Phylogeny

Objective To explore the significance in judging the different clinical stages of relapsing polychondritis (RP) patients through examining the changes of aggrecanase and metabolic fragments of aggrecan.MethodsIn comparison with the control group (20 cases),40 patients were divided into the stable stage group (22 cases) and the active stage group (18 cases).The aggrecanase-generated neoeptitopes in cartilage matrix were analysed by immunohistochemistry and Western blot(WB) respectively.The mRNA and protein levels of aggrecanase-1,2 expressed in cartilage cells were measured by real-time reverse transcriptional polymerase chain reaction(RT-PCR) and WB respectively.The difference of these results among these three groups was analyzed accordingly.ResultsThe expression of aggrecanase-1,2 in mRNA level was measured by real-time RT-PCR.The values of aggrecanase-1,2 mRNA 2 -ΔΔC1 were 1.00 ± 0.26 and 1.00 ± 0.27 in control group,1.47 ± 0.11 and 1.57 ± 0.13 in stable stage group,2.09 ±0.12 and 2.09 ± 0.19 in active stage group respectively.By one-way ANOVA analysis,the difference between every two groups was statistically significant (F was 299.113 and 459.013,P ＜ 0.01 ).In comparison with control group,aggrecanase-1,2 increased significantly in both stable and active stage group (P ＜ 0.01 ) and aggrecanase-1,2 increased more significantly in active stage group than in stable stage group (P ＜ 0.01 ).The results from WB analysis indicated that aggrecanase-1,2 could not be detected in control group,and they were detectable in stable stage group and increased in active stage group at the relative molecular of 68 000 Da or 73 000 Da respectively.The aggrecanase-generated neoeptitopes were analyzed by WB as well.The results indicated that NITEGE and ARGSV could be detected in stable stage group and increased in active stage group at the relative molecular of 70 000 Da or 48 000 Da respectively,but there were no signals in control group.Similar with the previous WB results,no signals of NITEGE or ARGSV eptitopes were detected in normal cartilage matrix ( no red staining) by use of immunohistochemical staining.However,in stable stage group and active stage group,these eptitopes were apparently detected (obviously red staining).ConclusionWith the progression of the RP,the activity of the aggrecanase is enhanced,and the degradation of the aggrecan is increased,associated with the severity of the disease.