Objective To study the biological characteristics of the human breast cancer cell mammospheres (MSs) ,and con‐struct breast cancer stem cell experiment model .Methods MCF‐7 cells were cultured in the serum‐free media supplemented with growth factors (the MCF‐7 group) ,and the MSs was collected (the MSs group) .The migration ,invasive and animal tumor forma‐tion abilities of MSs were detected by wound healing ,transwell invasive assay and animal tumor formation test .Results The wound line of MSs healed after 48 hours ,but the line of MCF‐7cells could not heal after 48 h .The number of the cells going through the membrane in MSs group was (76 .24 ± 0 .35) ,and the number in MCF‐7cells was (17 .38 ± 0 .18)(P<0 .05) .MSs had stronger ani‐mal tumor formation ability than MCF‐7 cells .Conclusion MSs have stronger abilities in migration ,invasive and animal tumor for‐mation ,and could be used in the studies of breast cancer stem cell as experimental model .

Objective To observe the effects of melatonin on synaptic plasticity impaired by spinal cord injury in rats. Methods A total of 54 female Sprague-Dawley rats were divided into sham group (n=18), control group (n=18) and melatonin group (n=18). Spinal cord inju-ry model was established with modified Allen's method at T10 (10 g from 25 mm height). The number of neurons and the expression of the Nissl body were detected with immunofluorescence and Nissl staining. The expression of neurofilament-200 (NF-200), brain-derived neuro-trophic factors (BDNF), Synapsin I and growth-associated protein-43 (GAP-43) was detected with Western blotting. Results Seven days af-ter injury, the number of motoneurons, the expression of Nissl body in motoneurons, and the expression of BDNF, Synapsin I and GAP-43 decreased in the control group compared with those in the sham group, and they increased in the melatonin group compared with those in the control group. Conclusion Melatonin can repair the impaired synaptic plasticity, which might promote the functional recovery after spi-nal cord injury.

OBJECTIVE:To investigate the compatibility stability of Fat-soluble vitamin (Ⅱ)/Water-soluble vitamin for in-jection with common electrolytes. METHODS:Referring to clinical common dose,Fat-soluble vitamin (Ⅱ) for injection/Wa-ter-soluble vitamin for injection collective packing [containing Fat-soluble vitamin(Ⅱ) for injection 2 ampoules and Water-soluble for injection 1 ampoule] were respectively mixed with Glucose injection,Potassium chloride injection,Concentrated sodium chlo-ride injection,Sodium bicarbonate injection,Potassium aspartate injection,Potassium aspartate and magnesium aspartate injection, Sodium glycerophosphate injection,Multi-trace elements injection (Ⅱ) to obtain mixture A-H. At room temperature (25 ℃),these mixtures were investigated in terms of appearance,pH value,osmotic pressure molar concentration and the number of insolu-ble particles at 0,1,2,3,4 h. The contents of bacterial endotoxin were tested at 0,4 h. RESULTS:Within 4 h after mixing, there was no significant change in appearance of those mixtures;pH value of mixture H changed greatly (RSD=5.13%,n=5), and that of mixture D and G increased significantly. The osmotic pressure molar concentration of those mixtures had no significantly change(RSD<2%,n=5)and lower than 600 mOsmol/kg. The bacterial endotoxin tests of those mixtures were negative. Two and four hours after mixing,the number of insoluble particles ≥10 μm in mixture B was increased significantly;2,3,4 h after mix-ing,the number of insoluble particles ≥10 μm in mixture E,F,H were increased significantly;0 ,1,2,3,4 h after mixing,the number of insoluble particles ≥10 μm in mixture G was increased significantly. There was statistical significance in the number of insoluble particle ≥10 μm in above mixtures compared to mixture A at the same time point(P<0.05),but it was in line with the standard of 2015 pharmacopeia. One,two,three and four hours after mixing,the number of insoluble particle ≥10 μm in mixture D was increased significantly,there was statistical significance compared to mixture A at the same time point(P<0.05);the num-ber of insoluble particle ≥10 μm in mixture D was beyond prescribed scope of pharmacopeia at 2,3,4 h after mixing. Within 4 h after mixing,both the number of insoluble particle ≥10 μm in mixture C and the number of insoluble particle ≥25 μm in edch mixture had no significant change,in accordance with pharmacopeia standard. CONCLUSIONS:Fat-soluble vitamin (Ⅱ)/Wa-ter-soluble vitamin for injection is not suitable for mixing with Multi-trace elements injection(Ⅱ),Sodium glycerophosphate injec-tion or Sodium bicarbonate injection due to great change of pH value and the number of insoluble particles.

This study aimed to investigate whether β-elemene could improve the dysfunction of vascular endothelial cells induced by low shear force (LSS), and the proliferation and migration of vascular smooth muscle cells induced by oxidized low-density lipoprotein (ox-LDL). Parallel plate flow chambers and ox-LDL were used to establish vascular endothelial cells (ECs) dysfunction model and vascular smooth muscle cell (VSMCs) proliferation and migration model, respectively, and the effects of β-elemene on ECs dysfunction and VSMCs proliferation and migration were examined. The activity of ROS in ECs was measured by DHE and the activity of NO in ECs was tested by DAF-FM DA. The protein phosphorylation of Akt and ERK in ECs were detected by Western blot. The proliferation of VSMCs was measured by MTT. The migration of VSMCs was examined by cell scratch test and Transwell assay. The gene expression of MMP-2 and MMP-9 in VSMCs was measured by RT-qPCR. In ECs, β-elemene could significantly reduce the LSS-induced increase in ROS, significantly increase the LSS-induced decrease in NO, decrease the phosphorylation of ERK, and increase the phosphorylation of Akt. In VSMCs, β-elemene could significantly reduce the proliferation and migration of VSMCs induced by ox-LDL, and reduce the gene expression of MMP-2 and MMP-9. To conclude, β-elemene can improve the LSS-induced ECs dysfunction and ox-LDL-induced VSMCs proliferation and migration.

Chronic myeloid leukemia (CML) is characterized by the accumulation of active BCR-ABL protein.Imatinib is the first-line treatment of CML;however,many patients are resistant to this drug.In this study,we aimed to compare the differences in expression patterns and functions of time-series genes in imatinib-resistant CML cells under different drug treatments.GSE24946 was downloaded from the GEO database,which included 17 samples of K562-r cells with (n=12) or without drug administration (n=5).Three drug treatment groups were considered for this study:arsenic trioxide (ATO),AMN107,and ATO+AMN107.Each group had one sample at each time point (3,12,24,and 48 h).Time-series genes with a ratio of standard deviation/average (coefficient of variation) ＞0.15 were screened,and their expression patterns were revealed based on Short Time-series Expression Miner (STEM).Then,the functional enrichment analysis of time-series genes in each group was performed using DAVID,and the genes enriched in the top ten functional categories were extracted to detect their expression patterns.Different time-series genes were identified in the three groups,and most of them were enriched in the ribosome and oxidative phosphorylation pathways.Time-series genes in the three treatment groups had different expression patterns and functions.Time-series genes in the ATO group (e.g.CCNA2 and DAB2)were significantly associated with cell adhesion,those in the AMN107 group were related to cellular carbohydrate metabolic process,while those in the ATO+AMN107 group (e.g.AP2M1) were significantly related to cell proliferation and antigen processing.In imatinib-resistant CML cells,ATO could influence genes related to cell adhesion,AMN107 might affect genes involved in cellular carbohydrate metabolism,and the combination therapy might regulate genes involved in cell proliferation.

The rapid mutation and widely spread of duck hepatitis A virus (DHAV) lead to the vast economic loss of the duck industry. To prepare and evaluate bivalent inactivated vaccine laboratory products of DHAV, 6 strains were screened from 201 DHAV-1 strains and 38 DHAV-3 strains by using serotype epidemiological analysis in most of the duck factory. Vaccine candidate strains were selected by ELD50 and LD50 tests in the 6 strains. Continuously passaged, the 5th passaged duck embryos bodies grinding fluid was selected as vaccine virus seeds. The virus seeds were treated with formaldehyde and water in oil in water (W/O/W) emulsions, making into three batches of two bivalent inactivated vaccine laboratory products. The safety test, antibody neutralization test, challenged protection and cross immune protection experiment suggested that the vaccines possessed good safety, and neutralizing antibodies were detected at 7th day and the challenged protection rate reached 90% to 100% at the 14th and 21st day. Moreover, immune duration of ducklings lasted more than five weeks. However, cross-immunity protection experiments with DHAV-SH and DHAV-FS only had 20%-30%. The two bivalent inactivated vaccine laboratory products of duck viral hepatitis were effective and reliable, providing a new method as well as a new product for DHAV prevention and control.
Animals ; Antibodies, Neutralizing ; blood ; Ducks ; virology ; Hepatitis Virus, Duck ; Hepatitis, Viral, Animal ; prevention & control ; virology ; Neutralization Tests ; Picornaviridae Infections ; prevention & control ; veterinary ; Poultry Diseases ; prevention & control ; virology ; Vaccines, Inactivated ; immunology ; Viral Hepatitis Vaccines ; immunology

Objective To determine the diagnostic value of shoulder MR arthrography in revealing the injuries of anterior glenoid labrum,and to compare MR arthrography with arm in neutral position with MR arthrography with arm in abduction and external rotation(ABER)position.Methods MR arthrography of the shoulder,including additional oblique axial sequences with the patient in the ABER position,was performed in 44 patients.The injuries of anterior glenoid labrum of these patients were retrospectively evaluated.The result was compared with that of arthroscopy.Results For displaying the abnormalities of anterior glenoid labrum,the sensitivity,specificity,and accuracy of axial MR images with arm in neutral position were 79.3%(23/29),100%(15/15),and 86.4%(38/44),respectively.While the results of ABER position oblique axial images were 93.1%(27/29),100%(15/15),and 95.5%(42/44), respectively.The difference of sensitivity between axial and ABER-position scans was statistically significant (P

OBJECTIVE To survey the qualification rate of sputum specimen.METHODS The specimen collecting and delivering time(morning,after deep coughing and gargling or not) was investigated.Aerobic bacteria isolated rate was evaluated.RESULTS The mean transportation time in positive aerobic bacteria isolated specimens and negative ones was 75 min and 124 min,respectively.Aerobic bacteria isolated rate was higher in sputum specimen that were microscopically screened for greater than 25 neutrophils,than in sputum specimen that were less then 25 neutrophils and greater than 10 BSE(buccal squamous epithelial) cells per 100? field.CONCLUSIONS Lower respiratory tract specimens should be delivered to the laboratory within 1 hour.Sputum specimen should be collected in the morning and after deep coughing and gargling.Microscopic examination should be mandatory in sputum microbiology,both for specimen evaluation and as a guide to what to look for in culture.

OBJECTIVE:To develop a method for rapid,accurate and simultaneous determination of dexzopiclone,zolpidam and zaleplon in human plasma. METHODS:After the plasma sample was processed by liquid-liquid extraction,UPLC-MS/MS was established to detect plasma sample with carbamazepine as internal standard. The separation was performed on a Waters ACQUITY UPLC HSS T3 column with mobile phase composed of 0.1% ammonium solution-acetonitrile (gradient elution) at flow rate of 0.2 ml/min. The column temperature was set at 40 ℃,and sample size was 5 μl. The detection was performed by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. The mass transition ion-pairs were as follows:m/z 308.2→263.1(zolpidem),m/z 389.3→245.0(dexzopiclone),m/z 306.2→236.1(zaleplon),m/z 237.3→151.2(internal standard). RE-SULTS:The linear range of zolpidem,dexzopiclone and zaleplon were 0.02-20.00,0.50-20.00,0.02-20.00 ng/ml,respectively (r=0.990 1,0.996 8,0.991 7). LLOQ of them were 0.02,0.50,0.02 ng/ml,and detection limit were 0.01,0.20,0.01 ng/ml. RS-Ds of intra-day and inter-day were all lower than 15% ;extraction recovery were 88.9% -106.5% ;matrix effect were 94.8%-106.3%. CONCLUSIONS:The method is simple,rapid,sensitive and specific,and it can be used for simultaneous deter-mination of zaleplon,zolpidem and dexzopiclone in human plasma.

Objective To study the cyclooxygenase (COX-2) ,nuclear factor-κB (NF-κB) and vascular endothelial growth factor (VEGF) expression and clinical significance in triple negative breast cancer .Methods From January 2010 to December 2014 ,breast cancer treatment in our hospital 100 patients for the study ,50 patients with triple negative breast cancer ,50 cases of non-triple neg-ative breast cancer was detected by immunohistochemistry ,100 cases of breast cancer were detected by immunohistochemistry in or-ganizations COX-2 ,NF-κB and VEGF expression of lymphatic vessel invasion and lymph vessel density and D2-40 mark detection , statistical analysis of relevant clinical and pathological information .Results COX-2 in triple negative and non-triple negative breast cancer were 76% ,70% ,the difference was not statistically significant (P>0 .05) ,VEGF triple negative breast cancer and non-triple negative breast lesions in cancerous lesions positive expression rates of 60% and 36% ,respectively ,which had significant difference (P<0 .05) .NF-κB in triple negative breast cancer lesions and non-triple negative breast lesions positive expression rate was 66%and 32% ,respectively ,which had significant difference (P< 0 .05) .Triple negative breast cancer NF-κB and VEGF ,COX-2 and VEGF expression was significantly positively related to breast cancer .Conclusion Radiation and chemotherapy is a major means of triple negative breast cancer postoperative treatment ,while inhibiting the NF-κB ,the expression of VEGF and COX-2 is expected to become the new target for treatment of triple negative breast cancer ,is worth exploring .