Peptidomic investigation of Neoponera villosa venom by high-resolution mass spectrometry: seasonal and nesting habitat variations

Background: Advancements in proteomics, including the technological improvement in instrumentation, have turned mass spectrometry into an indispensable tool in the study of venoms and toxins. In addition, the advance of nanoscale liquid chromatography coupled to nanoelectrospray mass spectrometry allows, due to its high sensitivity, the study of venoms from species previously left aside, such as ants. Ant venoms are a complex mixture of compounds used for defense, predation or communication purposes. The venom from Neoponera ants, a genus restricted to Neotropical regions, is known to have cytolytic, hemolytic, antimicrobial and insecticidal activities. Moreover, venoms from several Neoponera species have been compared and differences in their toxicity related to nesting habitat variation were reported. Therefore, the present study aimed to perform a deep peptidomic analysis of Neoponera villosa venom and a comparison of seasonal and nesting habitat variations using high-resolution mass spectrometry. Methods: Specimens of N. villosa ants were captured in Panga Natural Reserve (Uberlândia, MG, Brazil) from arboreal and ground-dwelling nests during summer and winter time. The venom glands were dissected, pooled and disrupted by ultra-sonic waves. The venom collected from different habitats (arboreal and ground-dwelling) and different seasons (summer and winter) was injected into a nanoACQUITY ULPC hyphened to a Q-Exactive Orbitrap mass spectrometer. The raw data were analyzed using PEAKS 7. Results: The results showed a molecular diversity of more than 500 peptides among these venoms, mostly in the mass range of 8004000 Da. Mutations and post-translational modifications were described and differences among the venoms were observed. Part of the peptides matched with ponericins, a well-known antimicrobial peptide family...

Helicobacter pylori Antigens Inducing Early Immune Response in Infants

To identify the Helicobacter pylori antigens operating during early infection in sera from infected infants using proteomics and immunoblot analysis. Two-dimensional (2D) large and small gel electrophoresis was performed using H. pylori strain 51. We performed 2D immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM) antibody immunoblotting using small gels on sera collected at the Gyeongsang National University Hospital from 4–11-month-old infants confirmed with H. pylori infection by pre-embedding immunoelectron microscopy. Immunoblot spots appearing to represent early infection markers in infant sera were compared to those of the large 2D gel for H. pylori strain 51. Corresponding spots were analyzed by matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS). The peptide fingerprints obtained were searched in the National Center for Biotechnology Information (NCBI) database. Eight infant patients were confirmed with H. pylori infection based on urease tests, histopathologic examinations, and pre-embedding immunoelectron microscopy. One infant showed a 2D IgM immunoblot pattern that seemed to represent early infection. Immunoblot spots were compared with those from whole-cell extracts of H. pylori strain 51 and 18 spots were excised, digested in gel, and analyzed by MALDI-TOF-MS. Of the 10 peptide fingerprints obtained, the H. pylori proteins flagellin A (FlaA), urease β subunit (UreB), pyruvate ferredoxin oxidoreductase (POR), and translation elongation factor Ts (EF-Ts) were identified and appeared to be active during the early infection periods. These results might aid identification of serological markers for the serodiagnosis of early H. pylori infection in infants.

Identification and characterization of a novel neuropeptide (neuropeptide Y-HS) from leech salivary gland of Haemadipsa sylvestris / 中国天然药物

The present study was designed to identify immunomodulatory components from the leech salivary gland of Haemadipsa sylvestris. The Sephadex G-50, Resource(TM) S column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify the salivary gland extracts (SGE). Structural analysis of isolated compounds was based on Edman degradation and matrix assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). The cDNA encoding the precursor of the compound was cloned from the cDNA library of the salivary gland of H. sylvestris. The levels of inflammatory mediators, including tumor necrosis factor-α (TNF-α), interferon γ (IFN-γ), interleukin-6 (IL-6), and monocyte chemotactic protein-1 (MCP-1) were assayed using an enzyme-linked immunosorbent assay (ELISA). The effects on cell proliferation and cell viability were observed using MTT assay. A novel neuropeptide Y (Neuropeptide Y-HS) from the leech salivary gland of H. sylvestris was purified and characterized. It was composed of 36 amino acid residues and the amino acid sequence was determined to be FLEPPERPAVFTSVEQMKSYIKALNDYYLLLGRPRF-NH2, containing an amidated C-terminus. It showed significant inhibitory effects on the production of inflammatory cytokines including TNF-α, IFN-γ, IL-6, and MCP-1. Neuropeptide Y was identified from leeches for the first time. The presence of neuropeptide Y-HS in leech salivary gland may help get blood meal from hosts and inhibit inflammation.

Screening of Serum Markers in Primary Immune Thrombocytopenia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the serum protein fingerprints of immune thrombocytopenia (ITP) patients and healthy controls by using weak cation exchange nanometer magnetic beads and MALDI-TOF-MAS technology, to identify the proteins with different expression, to establish the diagnostic model for ITP and to explore the pathogenesis of ITP.</p><p><b>METHODS</b>A total of 40 patients with ITP and 40 healthy controls were selected, the serum protein components were captured by using weak cation exchange nanaometer magnetic beads, the protein spectra of all specimens were detected by Autoflex II matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI- TOF-MS) and then the data were analyzed by CliprotoolsTM2.2 software, by which the distinct protein molecules were screened to set up ITP diagnostic model. To identify the established model, the sera of 20 ITP patients and 20 healthy controls were selected to make category and cross validations.</p><p><b>RESULTS</b>The detection of Clinprot system and the analysis of CliprotoolsTM2.2 software showed that about 55 protein peaks were detected with the range of 700 Da to 10 000 Da of molecular weight in the protein spectrum of serum speciments from 40 ITP patients and 40 healthy controls. Compared with healthy controls, 19 protein expression peaks with statistically significant difference were found in ITP patients (P < 0.05), among them 5 expressions were up-regulated and 14 expressions were down-regulated. The diagnostic model on basis of Supervised Neural Network Algorithm (SNN) was established through 10 MS peaks with strongest capability in ITP group and control group automatically distinguished by software, and it is expected that the sensitivity of model group reached to 100%, and the specificity to 100%. The category validation showed that this diagnostic model correctly identificed all 20 ITP patients and 20 healthy controls, and in cross validation, the model sensitivity was 100% and the specificity was 100%.</p><p><b>CONCLUSION</b>The ITP SNN model ertablished by using ChinProt System with high flax and good repetition is composed of 10 protein peaks with significant difference, this model can effectively distinguish ITP patients and healthy controls.</p>

Structural characterization and analysis of Omalizumab and its biosimilar CMAB007 by LC-MS / 生物工程学报

We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China.

Expression of prolyl 4-hydroxylase beta-polypeptide in non-small cell lung cancer treated with Chinese medicines / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the role of prolyl 4-hydroxylase beta polypeptide (P4HB) expressed in lung carcinoma and the intervention effect of Yiqi Chutan Formula (, YQCTF).</p><p><b>METHODS</b>Lung carcinoma model was established by subcutaneously inoculating LEWIS lung carcinoma cells in C57BL/6J mice. The differential expression of P4HB protein between the YQCTF (3.0 g/kg, gavage, once daily, 21 days) group and the control group was acquired by a 2 fluorescence difference gel electrophoresis (2D-DIGE), verified by Western blotting and identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS). The expression of P4HB and P4HB mRNA in cultured A549 cells from cisplatin (DDP) 1.5 µg/mL group and 15% serum combined with DDP 1.5 µg/mL group were detected by cellular immunohistochemistry and reverse transcription-polymerase chain reaction, respectively.</p><p><b>RESULTS</b>The proteomics research discovered that one-third of differential proteins including P4HB were decreased in the YQCTF group (P<0.01). Clinical pathology and tissue microarray studies showed that P4HB expression in lung cancer tissue was stronger than adjacent tissues and normal lung epithelial (P<0.01). In the YQCTF and DDP combined groups, the expression of P4HB and P4HB mRNA in A549 cell were decreased significantly (P<0.01).</p><p><b>CONCLUSION</b>YQCTF could inhibit the LEWIS lung carcinoma's growth, decrease the expression of P4HB in LEWIS lung carcinoma and A549 cells. YQCTF might take effect through regulating P4HB in endoplasmic reticulum to inhibit the incidence and growth process of lung carcinoma.</p>

Characterization and comparison of interferon reference standards using UPLC-MS / 药学学报

The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.

Application of surface enhanced laser desorption/ionization time-of-flight mass spectrometry in the diagnosis of leukemia / 中国实验血液学杂志

This study was purposed to find new biomarkers and to establish protein finger print model for diagnosis of leukemia. A total of 40 leukemia samples and 37 healthy control samptes were tested by surface enhance laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF- MS). The data of spectra were analyzed by bioinformatics tools like Biomarker Patterns 5.0 and discriminant analysis to establish diagnostic mode1. The results showed that 22 protein features were stably detected by protein fingerprint, The detective model combined with 3 biomarkers (m/z 4650, 8609 and 11660) could differentiate leukemia with sensitivity of 97.5% (39/40) and specificity of 91.9%(34/37). It is concluded that the detective model established by 3 protein features may be a novel method for diagnosis of leukemia.

N terminal sequencing for practical detection of monoclonal antibody / 生物工程学报

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.

Rapid and high-throughput identification of recombinant bacteria with mass spectrometry assay / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To construct a rapid and high-throughput assay for identifying recombinant bacteria based on mass spectrometry.</p><p><b>METHODS</b>Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques were used to identify 12 recombinant proteins (10 of Yersinia pestis, 1 of Campylobacter jejuni and 1 of Helicobacter pylori). A classification model for the various phase of recombinant bacteria was established, optimized and validated, using MALDI-TOF MS-ClinProTools system. The differences in the peptide mass spectra were analyzed by using Biotyper and FlexAnalysis softwares.</p><p><b>RESULTS</b>Models of GA, SNN, and QC were established. After optimizing the parameters, the GA recognition model showed good classification capabilities: RC=100%, mean CVA=98.7% (the CVA was 96.4% in phase 1, 100% in phase 2, 98.4% in phase 3, and 100% in phase 4, respectively) and PPV=95%. This model can be used to classify the bacteria and their recombinant, which only requires 3.7×103 cells for analysis. The total time needed is only 10 min from protein extraction to reporting the result for one sample. Furthermore, this assay can automatically detect and test 96 samples concurrently. A total of 48 specific peaks (9, 16, 9, and 14 for the four stages, respectively) was found in the various phase of recombinant bacteria.</p><p><b>CONCLUSION</b>MALDI-TOF MS can be used as a fast, accurate, and high-throughput method to identify recombinant bacteria, which provide a new ideas not only for recombinant bacteria but also for the identification of mutant strains and bioterrorism pathogens.</p>

Study on serum protein mass spectrometric characteristics of acute leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the serum protein differential expression in acute leukemia patients and healthy control by differential protein mass spectrometry.</p><p><b>METHODS</b>Serum proteins of 51 acute leukemia (AL) patients and 10 healthy donors were extracted from their peripheral blood. After removing high abundance protein, serum low abundance proteins were separated by two dimensional gel electrophoresis, the differences of serum proteins in AL patients and healthy human were identified. The protein spots with differential expression were cut out and then undergone bleaching, gel digestion and peptide extraction. The peptide mass fingerprint analysis was performed by using MALDI TOF/TOF MS. The protein database MSDB Masort retrieval program was used to evaluate the results.</p><p><b>RESULTS</b>Using Student's t test,19 statistically significant abnormal expression proteins in the serum of AL patients were found compared with the healthy controls (P < 0.05). The expression of α1-trypsin inhibitor (P < 0.01), prealbumin (P < 0.01), trypsin inhibitor (P < 0.01), apolipoprotein E (P < 0.01) and apolipoprotein A-Ⅳ (P < 0.01) decreased, while retinol binding protein (P < 0.05), globin HP2 (P < 0.05), serum lectin (P < 0.05), H factor homologue protein (P < 0.05) and serum amyloid A1 (P < 0.01) increased. Further stratified analysis found that high serum lectin expression in AL patient resulted in poor outcomes.</p><p><b>CONCLUSION</b>There are a variety of serum proteins with differential expression in peripheral blood of AL patients. The differential expression of serum lectin is related to the therapeutic effect. The differential expression of these proteins can be used as a new diagnosis marker or prognostic indicator for acute leukemia.</p>

Proteomic identification of human serum biomarkers associated with high altitude pulmonary edema / 中国应用生理学杂志

<p><b>OBJECTIVE</b>High altitude pulmonary edema (HAPE), a life-threatening disease, has no biological markers used for the routine prevention, diagnosis and treatment. The aim of this study was to identify serum proteins differentially expressed in patients with HAPE for discovering essential biomarkers.</p><p><b>METHODS</b>A complete serum proteomic analysis was performed on 10 HAPE patients and on 10 high altitude and 11 sea level healthy people as control using two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/ionization mass spectrometry and peptide mass fingerprinting. Finally, two most significantly changed proteins were validated by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Eight protein spots stained with differential intensity, respresenting 5 distinct proteins were identified in patients compared with healthy controls through analysis of these composite gels. Among them, four proteins, namely alpha 1-antitrypsin(alpha1-AT), Haptoglobin(Hp), apolipoprotein A-1 (apoA-1) and Complement C3 increased remarkably, while one protein, apolipoprotein A-IV (apoA-IV) decreased significantly. The variation of alpha1-AT and Haptoglobin, as detected by ELISA, was consistent with the results from proteomic analysis.</p><p><b>CONCLUSIONS</b>It is well known that Hp, alpha1-AT and complement C3 are associated with inflammation and apoA-1 and apoA-IV play important roles in lipid absorption, transport and metabolism. Therefore, the significant expression changes of Hp, alpha1-AT and complement C3 and apoA-1 and apoA-IV between HAPE patients and their corresponding healthy controls highlight the role of inflammatory response system and lipid metabolism system in the pathophysiology of HAPE.</p>

Observation of serum protein fingerprinting in primary liver cancer patients of different Chinese medical syndromes before and after interventional treatment / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore changes of serum protein fingerprinting in primary liver cancer (PLC) patients of different Chinese medical syndromes before and after interventional treatment detected by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS).</p><p><b>METHODS</b>Totally 154 PLC patients were assigned to 5 groups, i.e., Gan depression syndrome (GDS, 37 cases), Pi deficiency syndrome (PDS, 45 cases), dampness heat syndrome (DHS, 18 cases), blood stasis syndrome (BSS, 28 cases), yin deficiency syndrome (YDS, 26 cases). The mass spectra of serum proteins was analyzed by using SELDI-TOF-MS. Then the correlation between Chinese medical syndrome types and the mass spectra of serum proteins was explored before and after interventional treatment.</p><p><b>RESULTS</b>Compared with the healthy control group, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in DHS with M/Z being 5 710 Da, in YDS with M/ Z being 6 992 Da, while it was up-regulated in PDS with M/Z being 5 816 Da and in BSS with M/Z being 4 297 Da, showing statistical difference (P < 0.01). Compared with before intervention, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in PDS with M/Z being 5 816 Da, in DHS with M/Z being 5 710 Da in BSS with M/Z being 4 297 Da, while it was up-regulated in YDS with M/Z being 6 992 Da, showing statistical difference (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>There was statistical difference in changes of serum protein fingerprinting in PLC patients of different Chinese medical syndromes before and after interventional treatment.</p>

Diagnosis model of idiopathic thrombocytopenic purpura based on platelet differential proteome / 中国实验血液学杂志

This study was purposed to establish a new quick and simple diagnostic method with high sensitivity and good specificity for idiopathic thrombocytopenic purpura (ITP) and to evaluate its significance. 240 platelet lysates (from patients with ITP, leukemia, MDS, and healthy adults, each of 60 cases) were randomly assigned to training set (120 cases) or validation set (120 cases), all of them were detected by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS), in order to identify the differentially expressed protein, the diagnostic model was established by means of artificial neural network (ANN), and was validated by blind test with SPSS 17.0. The results showed that 5 marked proteins significantly differentially expressed (P < 0.01), m/z of highly expressed proteins were 2234.30, 3476.36, and 7526.29, m/z of low expressed proteins were 4990.02 and 5152.39, respectively. The sensitivity and specificity of diagnostic model were 80.6% and 77.3% respectively. The area under the ROC curve consisting of the output value of artificial neura1 network was 0.837. Efficacy of the model was validated by means of blinded test. It is concluded that the ANN model is useful for clinical diagnosis of ITP on the basis of platelet protein fingerprint spectrum.

Matrix-assisted laser desorption/ionization time of flight mass spectrometry in the genetic basis of systemic lupus erythematosus and the complicating kidney injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is necessary to develop some innovative methods to reveal and discover the novel (SLE)-related protein molecules. In the present study, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was employed to detect the differential expression of serum polypeptides in the patients with systemic lupus erythematosus (SLE) presenting with a family history or complicating with kidney injury so as to identify the proteins associated with the genetic factors and kidney injury in SLE.</p><p><b>METHODS</b>The subjects recruited were divided into four groups, that is, a group of SLE patients with both family history and kidney injury, a group of SLE patients with only kidney injury but no family history, a group of SLE patients with neither family history nor kidney injury, and a control group consisting of healthy volunteers. By adopting MALDI-TOF MS analysis, the serum samples obtained from the three groups of SLE patients were examined and compared with those from the control group; the categorized peptide fingerprint profile was established via the biological data collected from the samples.</p><p><b>RESULTS</b>The expression of protein with a m/z of 4207 Da increased significantly in SLE patients; the protein with a m/z of 2658 Da was expressed in all SLE patients; three proteins (with m/z of 1465, 5332, and 5900 Da respectively) were expressed in the SLE patients complicated with kidney injury and the protein with a m/z of 1943 Da was expressed in SLE patients with family history.</p><p><b>CONCLUSION</b>A number of differential proteins were successfully detected and identified through MALDI-TOF MS detection and these proteins may be associated with the genetic basis of SLE and the complicating kidney injury.</p>

Caracterización de factores asociados con comportamiento suicida en adolescentes estudiantes de octavo grado, en tres colegios bogotanos / Factor Characterization Associated with Suicidal Behavior in 8"' Grade Adolescent Students in Three Schools from Bogotá (Colombia)

Objetivos: Determinar la probabilidad de riesgo suicida y/o enfermedad mental y factores asociados en estudiantes de secundaria de tres colegios bogotanos. Métodos: Estudio de corte transversal con 309 adolescentes. Resultados: El promedio de edad fue de 13,83 ± 0,9 años, predominó el género femenino (58,6%) y el estrato socioeconómico 3 (68,3%). La probabilidad de riesgo para comportamiento suicida y/o síntomas mentales fue de 47,6%; 26,5% tuvo alguna manifestación suicida; 14,23% tuvo ideación suicida en los últimos tres meses; 3,55% tuvo intentos suicidas alguna vez en la vida, y 8,73% tuvo ideación suicida e intentos suicidas en los últimos tres meses. El riesgo de comportamiento suicida y/o enfermedad mental fue explicado conjuntamente por la depresión (OR = 27,9, IC95% = 3,5-223,1), la baja autoestima (OR = 11,8, IC95% = 2,5-56,5), la disfunción familiar severa (OR = 3,4, IC95% = 1,2-9,7), el sexo femenino (OR = 2,1, IC95% = 1,2-3,8) y la edad mayor o igual a 15 años (OR = 1,9, IC95% = 0,9-3,9). El maltrato psicológico seguido del abuso físico se asociaron con manifestación suicida y/o enfermedad mental, y la buena relación familiar, con menor probabilidad. Conclusión: La depresión, la baja autoestima, la disfuncionalidad familiar, el género femenino, la edad > 15 y la violencia intrafamiliar son factores asociados al riesgo suicida y/o enfermedad mental en adolescentes, y las buenas relaciones familiares se asocian con menor riesgo.

Objective: To establish the probability for suicide risk and/or mental disorders, together with related factors among high school students in 3 schools in Bogota. Methods: Cross sectional study of 309 adolescents. Results: The average age was 13.83 ± 0.9, female dominance (58.6%) and a 3rd socioeconomic stratum (68.3%). The suicidal risk behavioral probability and/or mental symptoms was 47.6%, 26.5% exhibited some suicide manifestations, 14.23% had experienced suicidal ideas in the last 3 months, 3.55% had had suicide attempts at least once in life, and 8.73% had suicidal ideas in the last 3 months with suicide attempts. The risk of suicidal behavior and /or mental disorders was explained jointly by depression (OR=27.9, 95% CI: 3.5-223. 1), low self-esteem (OR=11.8, 95% CI: 2.5-56.5), severe family dysfunction (OR=3.4, 95%CI 1.2-9.7), being female (OR=2.1, 95% CI: 1.2-3.8) and being 15 or older (OR=1.9, 95% CI: 0.967-3.9). Psychological abuse followed by physical mistreatment was associated with suicidal behavior and /or mental illness while good family relationships were associated to lower probability. Conclusion: Depression, low self-esteem, severe family dysfunction, female gender, older age (> 15) and domestic violence are risk factors associated with suicide and/or mental disorders in adolescents; good family relationships are associated with lower risk.

Correlation analysis between the exposure levels and the serum protein fingerprints in population exposure to silica / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the correlation between the exposure levels and serum protein fingerprints in population exposed to silica.</p><p><b>METHODS</b>Liquid chip time-of-flight mass spectrometry technology was used to investigate the serum profiles in control group (30 cases), group exposed to silica (30 cases), silicosis group (I stage, 25 cases) and suspected silicosis group (30 cases), and screen the differential expression proteins. The correlation between the levels of the differential expression proteins and the exposure levels was performed.</p><p><b>RESULTS</b>Five differential expression proteins were found among 4 groups, the expression of 5081 and 5066 proteins was upregulated, and the expression of 3954, 2021 and 1777 proteins was downregulated. There was no the correlation between the exposure levels and the peak with M/Z among those proteins.</p><p><b>CONCLUSION</b>the results of present investigation indicated there was no correlation between the exposure levels and protein/peptide peak.</p>

The oral microbial fingerprint on the greasy tongue coating of patients with chronic gastritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the microbial changes on the greasy tongue coating of the patients with chronic gastritis and to explore the formation mechanism of greasy tongue coating.</p><p><b>METHODS</b>Forty cases of tongue coating samples from patients with chronic gastritis were collected, 20 cases of greasy fur (as the greasy fur group), 20 cases of non-greasy fur (as the non-greasy fur group), and 20 cases of tongue coating samples from healthy subjects (as the healthy control group). Using 16S rRNA gene denatured gradient gel electrophoresis (DGGE) the microbial population of the tongue coating was detected. The DGGE fingerprint of the bacterium on the tongue coating was obtained. After digitalized principle component analysis (PCA) and partial least squares (PLS-DA) were performed.</p><p><b>RESULTS</b>The microorganism compositions are different in the greasy fur group, the non-greasy fur group, and the healthy control group. (1) There were five significantly different bands between the greasy fur group and the non-greasy fur group, with the accuracy of 97.5% in judging the model. There were 8 significantly different bands between the greasy fur group and the healthy control group, with the accuracy of 95.0% in judging the model. There was no obvious difference between the healthy control group and the non-greasy fur group. (2) The brightness of band 8 was higher in the greasy fur group than in the non-greasy fur group and the healthy control group. It may be a new species closely associated with the formation of greasy tongue coating. Results of the sequence showed its nearest neighbor was Moraxella catarrhalis, but with the similarity of 96.2%. The brightness of band 10 was sequenced as the healthy control group > the non-greasy fur group > the greasy fur group. Results of the sequence showed it had 100.0% similarity to Rothia mucilaginosa (stick-slip Ross strain).</p><p><b>CONCLUSIONS</b>The bacteria species on band 8 may have a close correlation with the formation of greasy fur of chronic gastritis, while the bacteria species on band 10 may have a close correlation with the formation of non-greasy fur. They indicated the microbial changes in the oral cavity may be one of the formation mechanisms for greasy tongue coating.</p>

Dingxin recipe ( ) prevents ischemia/reperfusion-induced arrhythmias via up-regulating prohibitin and suppressing inflammatory responses / 中国结合医学杂志

<p><b>OBJECTIVE</b>To identify the underlying mechanisms of the protective effects of Dingxin Recipe (: , DXR), a Chinese compound prescription that has been used clinically in China for more than 20 years, on ischemia/reperfusion (I/R)-induced arrhythmias in rat model.</p><p><b>METHODS</b>A total of 30 rats were randomly divided into three groups: sham group, I/R group, and DXR-pretreated I/R (DXR-I/R) group. Rats in the DXR-DXRI/R group were intragastrically administrated with DXR (12.5 g/kg per day) for consecutive 7 days, while rats I/in the sham and I/R groups were administrated with normal saline. Arrhythmias were introduced by I/R and electrocardiograms (ECG) were recorded. Two-dimensional (2-D) polyacrylamide gel electrophoresis and matrix-matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used to identify assisted differentially expressed proteins. Immunohistochemistry, real-time quantitative polymerase chain reaction (RQ-RQPCR), Western blot, and enzyme-linked immunosorbent assay (ELISA) were performed to analyze proteins PCR), obtained in the above experiments.</p><p><b>RESULTS</b>DXR significantly reduced the incidence and mean duration of ventricular tachycardia and ventricular fibrillation and dramatically decreased the mortality, as well as arrhythmia score, compared with those of the I/R group. Among successfully identified proteins, prohibitin (PHB) and heart fatty acid binding protein (hFABP) were up-regulated in DXR-pretreated I/R rats compared with those of the I/R rats. In addition, compared with the I/R group, the level of glutathione (GSH) was elevated accompanied by reduced expressions of interleukin-6 (IL-6) and neutrophil infiltration in I/R rats with DXR pretreatment.</p><p><b>CONCLUSIONS</b>DXR could alleviate I/R-induced arrhythmias, which might be related to increased expression of PHB. The enhanced expression of PHB prevented against the depletion of GSH and consequently inhibited apoptosis of cardiomyocytes. Furthermore, up-regulation of PHB might ameliorate I/R-induced cell death and leakage of hFABP by suppressing neutrophil infiltration and IL-6 expressions.</p>

Measurement of the amino acid sequence for the fusion protein FP3 with LC-MS/MS / 药学学报

The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.