A polydimethylsiloxane ( PDMS )/glass microfluidic chip consisting of a three-layer sandwich structure and three-parallel micro-electrode system was fabricated for the separation of polystyrene microspheres according to the particle size in high conductive solution by electric field produced by alternating current. The principle of electrokinetics of microspheres directional movement was investigated. The results showed that, when the applied voltage was 14 V at 100 kHz, the separation efficiency of 10 and 25 μm polystyrene microspheres was the best. Similarly, with a voltage of 10 V at 2 MHz, the separation efficiency of 5 and 25 μm polystyrene microspheres could achieve the highest value. Meanwhile, the voltage of 11 V at 1 MHz was suitable for the separation of 5 , 10 and 25 μm polystyrene microspheres with the separation efficiency of over 90%. At the same time, the formation of the laminar region in the middle of the electrode gap was the key role of microsphere separation.

Objective To investigate the expression of FHITmRNA and WWOXmRNA in human breast cancer tissues and its relation to clinicopathological and other molecular parameters. Methods With reference to the expression of ?-actin,the expression of FHITmRNA and WWOXmRNA was determined by reverse (transcription)-polymerase chain reaction(RT-PCR) in 51 breast cancer and adjacent breast tissue, and (semi-quantitative) analysis of band densities was performed. The protein expression of estrogen receptor(ER), progesterone receptor (PR), Her-2 gene in the 51 breast cancer lesions was detected by (immunohistochemical) method. Results FHITmRNA and WWOXmRNA expression was significantly different in 54 breast cancer tissue compared to adjacent breast tissue (P0.05); of FHITmRNA and WWOX mRNA was related to axillary lymph node metastasis (P

Objective To establish a stable HCV full-length genome replication cell model which is labeled with reporter gene and easyly to quantify intracellular HCV proteins and RNA level. Methodsneo gene was inserted into Luc-JC1 to make Luc-JC construct. Luc-JC RNA was obtained by in vitro transcription and then delivered into Huh7 cells by transfection. G418-resistant clones of Huh7 cells were obtained by selection. Clones of HCV full-length genome replication cell were confirmed by luciferase activity assay, Western blot and cleaveage of eYFP-MAVS by HCV NS3/4A protease. Then, HCV replication cell colonies were treated by different dose IFN-α in order to observe the change of luciferase activity, HCV protein and RNA level. Results At 3-4 weeks post-transfection, visible colonies were selected and stained by crystal violet. Luciferase activity and HCV NS3, NS5A protein were detected by luciferase activity assay and Western blot, respectively. Subcellular localization of eYFP-MAVS transferred from mitochondria to cytoplasms by cleavage of NS3/4A protease in cell colonies. Luciferase activity, HCV protein and RNA diminished obviously after IFN-α treatment. Conclusion A stable HCV full-length genome replication cell model labeled by reporter gene was successfully established and reporter activity can be used to indicate level of HCV proteins and RNA in cells. This cell model is a useful tool for the study on HCV pathogenesis and the screening of antiviral drugs.

ObjectiveTo establish a method for content determination of total iridoid compounds and baldrinal and 11-ethoxyviburtinal from Valerianae Jatamansi Rhizoma et Radix; To determine the contents of total iridoid compounds and baldrinal and 11-ethoxyviburtinal in Valerianae Jatamansi Rhizoma et Radix from three medicinal origins.Methods UV spectrophotometry was applied, 11-ethoxyviburtinal (cyclopentane-pyran-7-formaldehyde, 4-ethoxy methyl) was set as the reference substance, and the content of total iridoid compounds was determined at 288 nm. HPLC method was used to simultaneously determine the contents of baldrinal and 11-ethoxyviburtinal. The HPLC analysis was performed on a Phenomenex Luna C18 column (250 mm×4.6 mm, 5μm). The mobile phase was composed of acetonitrile-water in gradient elution at a flow rate of 0.95 mL/min. The detection wavelength was 288 nm and the column temperature was 30℃.Results The total iridoid compounds, baldrinal and 11-ethoxyviburtinal were in good linearity within the ranges of 2.088–14.616μg/μL, 74.88–224.64μg, and 41.6–249.6μg, respectively. This method was precise, and with good repeatability, stability and recovery rate.Conclusion The method is accurate, simple, rapid, which can be used for the quality control of Valerianae Jatamansi Rhizoma et Radix.

OBJECTIVE:To study the preventive effect of Lonicera japonica alcohol extract on mice with liver injury based on metabolomics method. METHODS:30 mice were equally randomized into a normal control (isometric normal saline) group,a model(isometric normal saline)group and a group of L. japonica alcohol extract(2 g/kg). The mice were given drugs by ig once a day for 14 consecutive days. On the 8th day of administration,the models were established by giving 0.2% dimethyl sulfoxide (DMN,10 ml/kg)ip once a day for 7 consecutive days. Gas chromatography-mass spectrum(GC-MS)was used to analyze 24 h total ions chromatogram of the urine sample on the 1st,3rd,5th and 7th day of administration of drugs and DMN. The change in endogenic small molecule metabolites in urine was observed. Principal component analysis was employed to explore the change in the metabolite chromatogram and underlying biomarkers in urine. RESULTS:The contour of the chromatogram changed to a largest extent 1 to 5 d after given DMN,but showed an obvious trend towards regression 7 days thereafter. DMN resulted in increase in the contents of 8-phenyl-8-azbicyclo-[4,3,0]non-3-ene-7,9-dione,2-(6-heptynyl)-1,3 dioxolane,bis-(O-methyloxime)-4-ketoglu-cose,and decrease in the contents of malonic acid,2-(4- chlorophenylthiomethoxyl)ethyl,tetrahydro-2-furanacetaldehyde,D-ga-lactose,erythro-pentonic acid and galacturonic acid,in endogenic small molecule metabolites in mouse urine,for which Lonicera japonica alcohol extract can improve that. CONCLUSIONS:Previous administration of L. japonica alcohol extract ig has preven-tive effect to some extent on the physiological and metabolic conditions of mice with liver injury induced by DMN.

Objective To evaluate laparoscopic radical resection of metachronous colorectal carcinoma.Methods A total of 13 patients with metachronous colorectal carcinoma undergoing laparoscopic resection in Department of Gastrointestinal Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology from January 2013 to December 2015 were analyzed retrospectively.Results The mean time of surgery was (156 ± 9) min.Tumors were located in the right hemicolon in 3 cases,in the transverse colon in one,in the left hemicolon in 2,in the sigmoid colon in four and in the rectum in 4.The mean blood loss was (66 ± 21) ml.There was no conversion to open surgery.Two patients were done with protective ileostomy.Postoperative gastrointestinal function recovery time was (2.5 ± 0.7) days.One postoperative intra-abdominal bleeding was successfully controlled laparoscopically.Posteperative length of hospital stay was (26.2 ± 2.9) days.The median follow-up was 12 months (5-30 months) with no cancer recurrence.Conclusions Laparoscopic radical resection of metachronous colorectal carcinoma has good curative effect,and high success rate in spite of previous history of laparotomy.

Objective To study the expression of BRMS1mRNA in human breast cancer tissues and their significance.Methods The expression of BRMS1mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) in 71 breast cancer tissues and adjacent breast tissues,12 patients with benign breast tumors and 12 patients with normal breast tissue,and semi-quantitative analysis of band densities was also performed.Results The expression of BRMS1mRNA in 71 patients with breast cancer and adjacent breast tissue was 0.378?0.046 and 0.918?0.044,respectively;the expression of BRMS1mRNA in 12 patients with benign breast tumors and 12 patients with normal breast tissue was 0.908?0.047 and 0.934?0.028 respectively.BRMS1mRNA expression was significantly lower in breast cancer tissue compared to adjacent breast tissue,benign breast tumors and normal breast tissue(P0.05),but was related to axillary lymph node metastasis and clinical stage(P

Objective To study the effect of HCV receptors′sequence on virus entry based on the two-dimensional structure and via tandem expression of HCV receptors on mouse hepatocytes.Methods The construced recombinant expression vectors pCDH-hLDLR-hSR-BⅠ-hCD81-GFP, pCDH-hLDLR-hCD81-hSR-BⅠ and pCDH-hCLDN-1-hOCLN-DsRed were cotransfected into 293FT cells with package vectors.The collected recombinant lentivirus expressing hCLDN-1-hOCLN was concentrated and attacked mouse hepatocytes.The transgenic mouse hepatocytes with tandem overexpression of CLDN-1 and OCLN were established after G418-selection.The transduced cells LSCCO/Hepa1-6 and LCSCO/Hepa1-6 were sorted via flow cytometry and puro-G418-selection after recombinant lentivirus expressing hLDLR-hSR-BⅠ-hCD81 and hLDLR-hCD81-hSR-BⅠattacked Hepa1-6 respectively.The infectivity of transduced mouse hepatocytes LSCCO/Hepa1-6 and LCSCO/Hepa1-6 to HCV was analyzed via direct-infection of serum-derived virus.Furthermore, the effect of HCV receptors′sequence on virus entry was studied.Results Both LSCCO/Hepa1-6 and LCSCO/Hepa1-6 enhanced HCV-cell binding.The transduced mouse hepatocytes LSCCO/Hepa1-6 had more HCV endocytosis.Conclusion SR-BⅠhas priority over CD81 in HCV entry in the early stage.

Anastomotic leakage is one of the common complications after rectal cancer surgery. Advances in the field of rectal surgery, such as introduction of total mesorectal excision, double-stapling reconstruction techniques, and minimally invasive surgery have improved oncologic outcomes and resulted in more favorable functional results, with a greater proportion of patients undergoing sphincter-preserving surgeries. Despite technical improvements, the incidence of anastomotic leakage has not decreased significantly. The incidence of anastomotic leakage is related to many factors, including patient-related factors, such as male sex, obesity, low score of nutrition risk screening, and Ⅲ-Ⅴ grade of ASA grading; disease-related factors, such as lower tumor location, tumor diameter > 3 cm, preoperative chemoradiotherapy and comorbidity; surgery-related factors, such as open or laparoscopic surgery, blood supply of anastomosis, tension of anastomosis, preventive stoma, duration of surgery, intraoperative blood loss, intraoperative events, and contamination, as well as selection and use of anastomotic device. Fully understanding the risk factors of anastomotic leakage are very important for reducing the occurrence of anastomotic leakage. For patients with risk factors, appropriate preventive measures should be implemented timely to reduce the risk of anastomotic leakage.

Objective To explore humam cytomegalovirus(HCMV) encoded microRNAs during latent infection in order to help study HCMV virology and latent infection mechanisms.Methods A model of HCMV latent infection via THP-1 cells infected with HCMV was constructed.Deep-sequencing was performed using high-resolution Solexa sequencing platform.The secondary structure of the newly sequenced miRNA was predicted by RNAFold bioinformatics software. Results HCMV encoded various miRNAs during latent infection, including miR-US25-2-3p, miR-US25-2-5p, miR-UL112, miR-US25-1, miR-UL22A and PC-5p-148467 with a predicted length of 25 bp, named hcmv-miR-US33as-5p.Conclusion HCMV can express many types of miRNAs during latent infection.