This study was designed to use iTRAQ technology coupled with 2D LC-MS/MS to study the comparative proteomics of different processing technology for pilose antler. 1015 proteins were identified with 2D LC combined with MOLDI TOF/TOF mass spectrometry. Comparative analysis with Protein Pilot (Version 4.5) revealed that 87 proteins were changed (P ≤ 0.05, the ratio of > 1.50 or < 0.60 as the threshold selection of difference proteins), of which 24 were up regulated and 33 were down regulated in the traditional frying process (TFP) compared with the fresh pilose antler (P ≤ 0.05). 7 significant different proteins (P ≤ 0.001), most of these significantly changed proteins were found to be involved in calcium ion binding and ATP binding associated with human healthy. Freeze drying with protective agent (FDP) (Trehalose) can improve the content of significantly different proteins (P ≤ 0.001) including Collagen alpha-1 (XII) chain (COL12A1) and Collagen alpha-1 (II) chain (COL2A1). The significant function involves in platelets activating, maintenance of spermatogonium, and disorder expression in tumor cells. The functional annotation by Hierarchical clustering and GO (gene ontology) showed that the main molecule functions of the proteins significantly changed in these processes were involved in binding (52.7%), catalytic (25.3%), structural molecule and transporter (6.6%).
Animals ; Antlers ; chemistry ; Chromatography, Liquid ; Collagen ; chemistry ; Down-Regulation ; Freeze Drying ; Gene Expression Regulation ; Proteomics ; Tandem Mass Spectrometry ; Technology, Pharmaceutical ; methods ; Up-Regulation

Objective To study the role of arrestin domain-containing protein 4( ARRDC4) in regulation of enterovirus 71(EV71) triggered innate IL-6 production and the underlying mechanism .Methods THP-1-derived macrophages (t-M?) were transfected with ARRDC4 specific siRNA and negative control siRNA .The expression and production of IL-6, replication and virus titer of EV71, and the activation of signaling pathway adaptors were analyzed with quantitative real -time PCR, ELISA and Western blot.Results Upon EV71 infection, ARRDC4 was upregulated.ARRDC4 silencing could enhance mRNA expression and production of IL-6, thus increasing the replication and virus titer of EV71.In ARRDC4 silenced t-M?, the activation of p-65,IκBα,ERK,JNK and p38 was promoted.Conclusion ARRDC4 promotes EV71-t riggered IL-6 production by enhancing the activation of NF-κB and MAPK signaling pathway to inhibit EV71 infection, contributing to positive regulation of anti-EV71 innate immune responses .

To study the chemical constituents of Cymbopogon citratus, isolation and purification of constituents were carried out on silica gel, Sephadex LH-20 and prepatative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Eight compounds were isolated and identified as 3beta-methoxy lanosta-9(11)-en-27-ol (1), 3beta-hydroxylanosta-9 (11)-en (2), (24S) -3beta-methoxylanosta-9(11), 25-dien-24-ol (3), 8-hydroxyl-neo-menthol (4), (2E)-3,7-dimethyl-2,7-octadiene-1, 6-diol (5), (+)-citronellol (6), 7-hydroxymenthol (7) and ethyl nonadecanoate(8). Compounds 1 is a new one. Compounds 2-3 are obtained from C. citratus for the first time.
Cymbopogon ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry

OBJECTIVETo evaluate the safety and the clinical value of external use of jiuyi Powder (JP) in treating plasma cell mastitis using partial least-squares discriminant analysis (PLSDA).

METHODSTotally 50 patients with plasma cell mastitis treated by external use of JP were observed and biochemical examinations of blood and urine detected before application, at day 4 after application, at day 1 and 14 after discontinuation. Blood mercury and urinary mercury were detected before application, at day 1, 4, and 7 after application, at day 1 and 14 after discontinuation. Urinary mercury was also detected at 28 after discontinuation and 3 months after discontinuation. The information of wound, days of external application and the total dosage of external application were recorded before application, at day 1, 4, and 7 after application, as well as at day 1 after discontinuation. Then a discriminant model covering potential safety factors was set up by PLSDA after screening safety indices with important effects. The applicability of the model was assessed using area under ROC curve. Potential safety factors were assessed using variable importance in the projection (VIP).

RESULTSUrinary β2-microglobulin (β2-MG), urinary N-acetyl-β-D-glucosaminidase (NAG), 24 h urinary protein, and urinary α1-microglobulin (α1-MG) were greatly affected by external use of JP in treating plasma cell mastitis. The accuracy rate of PLSDA discriminate model was 74. 00%. The sensitivity, specificity, and the area under ROC curve was 0. 7826, 0. 7037, and 0. 8084, respectively. Three factors with greater effect on the potential safety were screened as follows: pre-application volume of the sore cavity, days of external application, and the total dosage of external application.

CONCLUSIONSPLSDA method could be used in analyzing bioinformation of clinical Chinese medicine. Urinary β2-MG and urinary NAG were two main safety monitoring indices. Days of external application and the total dosage of external application were main factors influencing blood mercury and urine mercury. A safety classification simulation model of treating plasma cell mastitis by external therapy of JP was established by the two factors, which could be used to assess the safety of external application of JP to some extent.

Acetylglucosaminidase ; Alpha-Globulins ; Discriminant Analysis ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Least-Squares Analysis ; Mastitis ; drug therapy ; Plasma Cells ; ROC Curve ; Safety

OBJECTIVESTo investigate mechanism for the increasing level of serum vascular endothelial growth factor(VEGF) in tumour patients during radiotherapy and the inhibitory action of the antisense oligodeoxynucleotide (AS-ODN) to the expression of VEGF protein by radiotherapy in the prostate cancer cell line (PC3M).

METHODSTo observe the changes of serum VEGF in the prostate cancer patients during radiotherapy dynamically and the inhibitory action of the antisense oligodeoxynucleotide to the expression of VEGF by radiotherapy in PC3M.

RESULTSThe changes of serum VEGF in three patients receiving radiotherapy had been observed continuously. The levels of serum VEGF began to increase when the patients received radiotherapy and rised up to peak value after fifteen days, then declined to the range of pre-radiotherapy. Irradiating the PC3M cells with X-rays significantly increased the VEGF expression and secretion. The expression of VEGF protein in the group treated by VEGF AS-ODNs and X-ray irradiation decreased significantly than the group treated only by X-ray irradiation.

CONCLUSIONSThe induction of VEGF protein expression by X-ray irradiation in tumor cells may result in the increasing of the VEGF in the prostate cancer patients during radiotherapy and the induction can be blocked by VEGF AS-ODNs.

DNA, Antisense ; pharmacology ; Endothelial Growth Factors ; antagonists & inhibitors ; blood ; genetics ; Gene Expression ; drug effects ; radiation effects ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; genetics ; Lymphokines ; antagonists & inhibitors ; blood ; genetics ; Male ; Prostatic Neoplasms ; blood ; pathology ; Radiotherapy ; adverse effects ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

The related substances in docetaxel injection were identified by LC-MS/MS. Ethyl acetate was used to extract the injection to remove the pharmaceutical excipients. HPLC separation was carried out on a Hedera ODS-2 column (150 mm x 2.1 mm, 5 microm) with a mobile phase consisting of acetonitrile - 0.1% acetate acid aqueous solution (40: 60). Electrospray ionization source was set in the positive mode for the LC-ESI-MS/MS, and the ion monitoring modes were full scan and product ion scan. According to the mass spectra of the related substances, the fragment profiles were explained, and the chemical structures were elucidated. Docetaxel and its main related substances were well separated. Nine related substances in docetaxel injection were detected by LC-MS/MS. Their chemical structures were proposed, and four of them were identified in the docetaxel injection for the first time. The established LC-MS/MS method is effective in the separation and identification of the related substances in docetaxel injection. The test results are useful for its quality control.
Antineoplastic Agents ; administration & dosage ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; Drug Contamination ; Injections ; Molecular Weight ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; Taxoids ; administration & dosage ; analysis ; chemistry

·Corneal confocal microscopy can be used in the real-time, noninvasive, high-resolution corneal structure at the cellular level, which has been widely used in corneal disease research. This review summarizes recent advances in corneal confocal microscopy in the study of infectious keratitis, dry eye, keratoconus, diabetic peripheral neuropathy, and clinical studies to guide corneal transplantation.

Objective To investigate the application effect of swallowing training and ice stimulation combined with electromyography biofeedback therapy in stroke patients with swallowing and coughing .Methods Seventy-nine stroke swallowing and coughing patients treated were collected and were randomly divided into two groups (39 cases in control group,40 cases in observation group ) according to the digital table .The control group was treated with normal swallowing training,while the observation group was treated with ice stimulation combined with electromyography biofeedback therapy.The clinical intervention effect and the VFSS results before and after the intervention were observed and compared .The complications of the two groups were statistically analyzed .Results The total effective rate of the observation group [90.00％(36/40)] was higher than that of the control group [71.79％(28/39),χ2 =4.255,P=0.039].After eight-week intervention,the VFSS score of the observation group was higher than that of the control group(t=12.173,P=0.000);and the incidence rate of complications of the observation group [7.50％(3/40)] was lower than that of the control group [25.64％(10/39),χ2 =4.727,P=0.030].Conclusion Swallowing training and ice stimulation combined with electromyography biofeedback therapy can significantly improve the swallowing function of stroke patients with swallowing and coughing ,and it is safe .

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

Objective To understand the relationship of tendency on drug abuse and related influencing factors for middle school students.Methods A total of 603 students wcre assessed with self-made copying style scale,consciousness to resistance of smoking,drinking and resistance ability through a questionnaire.A relationship model was set up by structure equation modeling.Results Copying style and resistance ability had significantly direct influence on tendency of drug abuse(with standardized coefficients were 0.202 and 0.092 respectively).Consciousness to resistance of smoking and drinking had significantly indirect influence on tendency of drug abuse.The goodneSS of fit for the structural equation model was satisfactory,with GFI,AGFI,RMSEA as 0.981,0.972 and 0.014 respectively.Conclusion It Was meaningful to grasp some copying skills and resistance ability to lower the tendency of drug abuse in middle school students.