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

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

OBJECTIVETo explore the active ingredients in the Chinese yellow wine could inhibit the proliferation and migration of rat vascular smooth muscle cells induced by homocysteine (Hcy).

METHODSThe primary culture and identification of rat vascular smooth muscle cells (VSMCs) was conducted, and the VSMCs in passage 4-7 were used in the following experiments. The VSMCs were divided into 7 groups: control, Hcy (1 mmol/L), Hcy + oligosaccharide, Hcy + polypeptides, Hcy + polyphenols, Hcy + alcohol, Hcy + Chinese yellow wine and were given the corresponding treatment. The proliferation of VSMCs was determined by MTT. Transwell chambers and would healing were employed to test the migratory ability of VSMCs. Wester blot and gelatin zymography were used to investigate the expressions and activities of metal matrix proteinase 2/9 (MMP-2/9) and tissue inhibitor of metalloproteinase 2 (TIMP-2) in VSMCs of each group.

RESULTSCompared with control group, the proliferation, migration and the expression and activity of MMP-2/9 of VSMCs were significantly increased in the VSMCs of Hcy group (P < 0.01). Compared with Hcy group, the proliferation, migration and the expression and activity of MMP-2/9 of VSMCs were significantly decreases in the VSMCs of polypeptides group, polyphenols group and Chinese yellow wine group. However, the expression of TIMP-2 among each group had no significant difference.

CONCLUSIONPolypeptides and polyphenols in the Chinese yellow wine could inhibit the proliferation and migration of VSMCs induced by Hcy.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Homocysteine ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Peptides ; chemistry ; Polyphenols ; chemistry ; Rats ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Wine

OBJECTIVETo evaluate the middle term effectiveness of medial patellofemoral ligament (MPFL) reconstruction for the treatment of recurrent patellar dislocation.

METHODSFrom February 2007 to January 2010, 65 patients including 6 males and 59 females with recurrent patellar dislocation received the MPFL reconstruction. The reconstruction was performed using ipsilateral semitendinosis tendon to restore the damaged MPFL. Patients were evaluated pre-operatively and post-operatively by physical and subjectively with the IKDC (International Knee Documentation Committee), Tegner, and Lysholm questionnaires and radiographic examination.

RESULTSThe average follow-up duration was 20 months (ranged, 15 to 23 months). No recurrent episodes of dislocation or subluxation occurred. A firm endpoint to lateral patellar translation was noted in all patients at most recent follow-up. The Lysholm subjective knee evaluation score improved from (60.6 +/- 3.7) preoperatively to (89.8 +/- 4.6) postoperatively; and Tegner scores improved from (3.6 +/- 0.4) to (5.6 +/- 0.3), IKDC from (40.0 +/- 3.5) to (82.0 +/- 3.6). Radiographic evaluation demonstrated improvements in the congruence and sulcus femoral angles.

CONCLUSIONMPFL reconstruction is an effective surgical procedure for the treatment of recurrent patellar dislocation.

Adolescent ; Female ; Femur ; surgery ; Humans ; Internal Fixators ; Ligaments ; surgery ; Male ; Patella ; surgery ; Patellar Dislocation ; surgery ; Reconstructive Surgical Procedures ; Young Adult

OBJECTIVETo explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.

METHODAcute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.

RESULTThe purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.

CONCLUSIONASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.

Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Neoplastic Stem Cells ; cytology ; drug effects ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects

OBJECTIVETo analyze the genetic polymorphism, distribution of haplotypes, common and well-documented (CWD) and rare alleles of high-resolution HLA-A, B and DRB1 alleles by analysis from hematopoietic stem cell (HSC) donors in Jiangsu Han Chinese.

METHODSPCR-sequence-based typing and PCR-sequence specific oligonucleotide probes methods were applied for HLA-A, B and DRB1 high-resolution genotyping of 3238 unrelated healthy donors of hematopoietic stem cells in Jiangsu branch of Chinese National Marrow Donor Program registry.

RESULTS46 alleles of HLA-A,85 HLA-B and 51 HLA-DRB1 locus were found. The frequencies of the most common alleles were A * 11:01 (16.52%), B * 13:02 (11.60%) and DRB1 *07:01 (15.78%). That of the most common haplotype was A * 30: 01-B * 13: 02-DRB1 * 07: 01 (8.87%). 40 alleles of HLA-A,77 alleles of HLA-B, and 47 HLA-DRB1 alleles of HLA-DRB1 were CWD, which account for 99. 8% of total number of samples, and a few rare alleles not reported in Chinese population were found.

CONCLUSIONThe results of high-resolution, CWD and rare alleles showed the characteristics of HLA distribution in Jiangsu Han population, which may be useful for finding HLA matched unrelated donors, as well as for HLA correlation with population genetics and disease association studies.

Alleles ; Asian Continental Ancestry Group ; genetics ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DRB1 Chains ; genetics ; Haplotypes ; Hematopoietic Stem Cells ; Humans ; Tissue Donors

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