Animals ; Haptoglobins ; metabolism ; Mesenteric Artery, Superior ; metabolism ; Mesenteric Vascular Occlusion ; blood ; Rabbits ; Serum ; metabolism

OBJECTIVETo explor the changes of serum proteomics in rabbits superior mesenteric artery occlusion (SMAO) shock as well as its possible effect in SMAO shock.

METHODSSMAO shock model in rabbits were induced by occlusion of the superior mesenteric artery, serum samples were obtained from rabbits before and after SMAO shock, proteins in samples were separated by two-dimensional electrophoresis(2-DE), spots in the 2-DE map were detected and evaluated by PDQuest software 8.0. The spots with different expression level were subjected to matrix assisted laser desorption/ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-TOF-MS) for identification, the protein database was searched to further characterized the differential proteins.

RESULTS19 differential protein spots were screened out in the 2-DE maps, 11 proteins were up-regulated and 8 proteins were down-regulated in SMAO shock rabbits' s serum. 4 of the 19 differential protein spots were selected for MALDI-TOF-TOF-MS study, and 2 of the 4 differential protein spots were identified satisfactoryly as paraoxonase and haptoglobin, which content were increased in rabbits' s serum after SMAO shock.

CONCLUSIONSerum proteomics of rabbit change remarkablely before and after SMAO shock, paraoxonase and haptoglobin may be associated with the compensation after SMAO shock.

Animals ; Arterial Occlusive Diseases ; blood ; complications ; Aryldialkylphosphatase ; metabolism ; Blood Proteins ; metabolism ; Female ; Haptoglobins ; metabolism ; Male ; Mesenteric Artery, Superior ; pathology ; Proteome ; metabolism ; Proteomics ; methods ; Rabbits ; Shock ; blood ; etiology

OBJECTIVETo search differentially expressed proteins in serum of patients with Crohn disease.

METHODSSerum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTSThe 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.

CONCLUSIONUp-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.

Adult ; Blood Proteins ; metabolism ; Case-Control Studies ; Crohn Disease ; blood ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins ; metabolism ; Humans ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the effect of Hyper-IL-6 on the proliferation of L-02 cells.

METHODSThe recombinant lentiviral vectors encoding Hyper-IL-6 (FIV-Hyper-IL-6), IL-6 (FIV-IL-6) and the lentiviral (FIV) were prepared using the FIV-based lentiviral vectors and the packaging system. The titer of FIV-Hyper-IL-6, FIV-IL-6 and FIV was tested with puromycin. L-02 cells were infected with FIV-Hyper-IL-6, FIV-IL-6, FIV, or mock-infected. The growth rate of L02 cells was analyzed with MTT at different time points after infection. The changes of Haptoglobin in L-02 cells were analyzed with RT-PCR.

RESULTSFIV-Hyper-IL-6, FIV-IL-6 and FIV were successfully constructed, and the titer was 107 pfu/ml. 48 hours after infection, the absorbance of the cells infected with FIV-Hyper-IL-6 was 0.6267+/-0.0256, and the absorbances of FIV-IL-6 infected cells, FIV infected cells and mock-infected cells were 0.5563+/-0.0112, 0.5040+/-0.0078 and 0.4790+/-0.0201, respectively. There were significant differences between the FIV-Hyper-IL-6 group and the other groups (F = 41.09, P less than 0.01). The ratio of the absorbance between haptoglobin mRNA and beta-actin was 0.7030+/-0.0106, 0.3355+/-0.0093, 0.1145+/-0.0076 and 0.1143+/-0.0153, respectively, in FIV-Hyper-IL-6 infected cells, FIV-IL-6 infected cells, FIV infected cells and mock-infected cells. There were significant differences between the FIV-Hyper-IL-6 group and the others (q = 57.5007, P less than 0.01).

CONCLUSIONCompared with IL-6, Hyper-IL-6 is more potent to stimulate proliferation and induce the expression of Haptoglobin in L-02 cells.

Cell Proliferation ; Genetic Vectors ; Haptoglobins ; genetics ; metabolism ; Hep G2 Cells ; Hepatocytes ; cytology ; metabolism ; Humans ; Interleukin-6 ; genetics ; metabolism ; Lentivirus ; genetics ; RNA, Messenger ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors ; Transfection

Advancements in the field of proteomics have provided great opportunities for the development of diagnostic and therapeutic tools against human diseases. In this study, we analyzed haptoglobin and amyloid A protein levels of vivax malaria patients with combinations of depletion of the abundant plasma proteins, 2-dimensional gel electrophoresis (2-DE), image analysis, and mass spectrometry in the plasma between normal healthy donors and vivax malaria patients. The results showed that the expression level of haptoglobin had become significantly lower or undetectable in the plasma of vivax malaria patients due to proteolytic cleavage when compared to healthy donors on 2-DE gels. Meanwhile, serum amyloid A protein was significantly increased in vivax malaria patient's plasma with high statistical values. These 2 proteins are common acute phase reactants and further large scale evaluation with a larger number of patient's will be necessary to establish the possible clinical meaning of the existential changes of these proteins in vivax malaria patients. However, our proteomic analysis suggests the feasible values of some plasma proteins, such as haptoglobin and serum amyloid A, as associating factor candidates for vivax malaria.
Blood Proteins/analysis/diagnostic use ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins/analysis/diagnostic use/*metabolism ; Humans ; Malaria, Vivax/diagnosis/*metabolism/parasitology ; Plasmodium vivax/physiology ; Proteomics/*methods ; Serum Amyloid A Protein/analysis/diagnostic use/*metabolism

Successful hematopoietic stem cell transplantation (HSCT) involves the restoration of hematopoietic function after engraftment, arising from the differentiation and proliferation of hematopoietic stem cells. Several factors could influence the course of allogeneic-HSCT (allo-HSCT). Therefore, knowledge of serum proteome changes during the allo-HSCT period might increase the efficacy of diagnosis and disease prevention efforts. This study conducted proteomic analyses to find proteins that were significantly altered in response to allo-HSCT. Sera from five representative patients who underwent allo-HSCT were analyzed by 2-dimensional gel electrophoresis and liquid chromatography tandem mass spectrometry, and were measured on a weekly basis before and after allo-HSCT in additional 78 patients. Fourteen protein spots showing changes in expression were further examined, and most proteins were identified as acute phase proteins (APPs). Studies of 78 additional patients confirmed that C-reactive protein (CRP) and haptoglobin undergo expression changes during allo-HSCT and thus may have the potential to serve as representative markers of clinical events after allo-HSCT. Maximal CRP level affected the development of major transplant-related complications (MTCs) and other problems such as fever of unknown origin. Particularly, an increase in CRP level 21 days after allo-HSCT was found to be an independent risk factor for MTC. Maximal haptoglobin and haptoglobin level 14 days after allo-HSCT were predictive of relapses in underlying hematologic disease. Our results indicated that CRP and haptoglobin were significantly expressed during allo-HSCT, and suggest that their level can be monitored after allo-HSCT to assess the risks of early transplant-related complications and relapse.
Adolescent ; Adult ; Biological Markers ; C-Reactive Protein/*metabolism ; Female ; Haptoglobins/*metabolism ; Hematopoietic Stem Cell Transplantation/*adverse effects ; Humans ; Male ; Middle Aged ; Proteome/*metabolism ; Proteomics ; Transplantation Conditioning ; Transplantation, Homologous ; Young Adult

OBJECTIVE: To investigate the expression of haptoglobin in the lesions of condyloma acuminatum (CA) at the mRNA and protein level, and to explore its role in the pathogenesis of CA. METHODS: The expressions of haptoglobin protein and mRNA in the skin tissues of 30 patients with CA and 20 normal controls were detected by immunohistochemistry(IHC), Western blot, and hybridization in situ. RESULTS: The in situ hybridization study showed that haptoglobin mRNA was expressed in the epidermal cells in the lesions of CA. The distribution of haptoglobin mRNA expression in the lesions of CA was similar to that of the normal controls, and the expression of haptoglobin mRNA in CA was higher than that of the normal controls. There was a significant difference in the positive expression of haptoglobin mRNA between the CA group and the control group (P<0.05). The immunohistochemical study showed that haptoglobin protein was expressed in the whole layers of epidermal keratinocytes in the lesions of CA at a high level and stronger staining was seen in the stratum basale and stratum spinosum. Haptoglobin protein was expressed predominantly in the stratum basale in normal skin tissues, while weak staining was seen below the stratum spinosum.There was a significant difference in the mean gray value between the CA group and control group (P<0.05). Western blot showed that the haptoglobin expression in CA lesions significantly increased compared with the normal skins (P<0.05). CONCLUSION The expression of haptoglobin mRNA in the CA lesions obviously increases and the epidermal cells in the CA lesions are able to synthesize haptoglobin protein. Haptoglobin in the CA lesions may involve in the local immunity escape by preventing Langerhans cell functional maturation and inhibiting the immunocompetence of keratinocyte.
Adolescent ; Adult ; Aged ; Blotting, Western ; Case-Control Studies ; Condylomata Acuminata ; genetics ; metabolism ; Epidermal Cells ; Haptoglobins ; genetics ; metabolism ; Humans ; In Situ Hybridization ; Keratinocytes ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Young Adult

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were rally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.
Aflatoxin B1/toxicity ; Animals ; Blood Proteins/drug effects/*metabolism ; Enzyme-Linked Immunosorbent Assay ; Haptoglobins/drug effects/*metabolism ; Immunoglobulins/*blood/drug effects ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred Strains ; Rats ; Rats, Wistar ; Trichothecenes/*toxicity ; Zearalenone/toxicity

3T3-L1 adipocytes express the B-cell-activating factor (BAFF) and three different BAFF receptors (BAFF-Rs). Furthermore, BAFF expression is regulated by inflammatory modulators, such as tumor necrosis factor-alpha and rosiglitazone. Here we investigated the function of BAFF in 3T3-L1 adipocytes and RAW 264.7 macrophages. We examined adipokine expression in 3T3-L1 adipocytes treated with 10 ng ml-1 BAFF. We also examined inflammatory molecule expression in RAW 264.7 macrophages treated with 10 or 100 ng ml-1 BAFF. We examined BAFF expression in the coculture of 3T3-L1 adipocytes and RAW 264.7 macrophages, as well as in white adipose tissue (WAT) of diet-induced obese (DIO) mice. We found that BAFF decreases leptin and adiponectin expression, but increases the expression of proinflammatory adipokines monocyte chemotactic protein-1, interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and haptoglobin. Coculturing the two cell types resulted in increased BAFF mRNA and protein expression, as well as modulation of BAFF-R mRNA expression in both cell types. These data indicate that BAFF might mediate adipocyte and macrophage interaction. When RAW 264.7 macrophages were treated with BAFF, BAFF-R expression was modulated as in coculture, and nitric oxide synthase and IL-6 expression increased. BAFF expression also increased in WAT of DIO mice. We propose that BAFF can regulate adipokine expression and possibly mediate adipocyte and macrophage interaction.
3T3-L1 Cells ; Adipocytes/drug effects/*metabolism ; Adipokines/genetics/*metabolism ; Adiponectin/genetics/metabolism ; Animals ; B-Cell Activating Factor/*metabolism/pharmacology ; Chemokine CCL2/genetics/metabolism ; Coculture Techniques ; Gene Expression Regulation/drug effects ; Haptoglobins/genetics/metabolism ; Inflammation Mediators/metabolism ; Interleukin-6/genetics/metabolism ; Leptin/genetics/metabolism ; Macrophages/drug effects/*metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; RNA, Messenger/genetics/metabolism

OBJECTIVETo compare the 2-DE profiles for serum proteins of different pathological stages during hepatocarcinogenesis.

METHODSSera from hepatocellular carcinoma patients, cirrhosis patients, chronic hepatitis patients and healthy controls were collected. After sonication, albumin and immunoglobulin (IgG) depletion, and desalination, sera were subjected to 2-DE, the differential protein spots were identified by MALDI-TOF-MS. Western blot was used to validate these differentially expressed proteins.

RESULTS2-DE sera protein profiles were obtained from the patient suffering from HCC, liver cirrhosis, chronic hepatitis, healthy controls in each group. From optimized 2-DE gel images of the above groups, 96 protein spots with more than 2-fold difference in intensity between the two groups were selected by image master 6.0 software, differential proteins including haptoglobin, SAA1 and SP40 were identified by MALDI-TOF-MS/MS. 7 different spots within more than 30 protein spots belonged to the same haptoglobin family. The differential expression of haptoglobin was confirmed by western blot.

CONCLUSIONSFour protein expression patterns have been identified during the pathological stages of hepatocarcinogenesis. Haptoglobin is significantly increased from liver cirrhosis to HCC. It implies that haptoglobin might be a potential biomarker in the early diagnosis of liver cancer.

Adult ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Blotting, Western ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Haptoglobins ; analysis ; Hepatitis, Chronic ; blood ; pathology ; Humans ; Liver Cirrhosis ; blood ; pathology ; Liver Neoplasms ; blood ; pathology ; Male ; Middle Aged ; Proteome ; metabolism ; Proteomics ; methods ; Young Adult