OBJECTIVETo analyze differentially expressed metastasis-associated proteins in Adenoid cystic carcinoma cell lines of human salivary gland by proteomics.

METHODSProtein expression alterations of ACC-2 and ACC-M cells were described by 2-D gels. After image analysis by software, proteins of interest were excised from the gels and identified by matrix assisted laser desorption ionization time-of-flight mass spectrometer.

RESULTS12 protein spots showed significantly differential expression patterns between two cell lines. In the identified protein candidates, transketolase, modulator recognition factor 2, Dim1p homolog, splicing factor (arginine/serine-rich 9) and v-Ha-ras l oncogene were all lowly expressed in the poorly metastatic ACC-2 cell and significantly upregulated in highly metastatic ACC-M cell, while type I collagen pro alpha and tumor necrosis factor (ligand) superfamily member 4 showed a high expression in ACC-2 cells and a low expression in ACC-M cells. Pirin (spot 6) just appears in ACC-2 cell and was not detectable in ACC-M cell, while retinal homeobox protein was just detected in ACC-M cell and did not appear in ACC-2 cell.

CONCLUSIONSThe proteins may be involved in the adenoid cystic carcinoma lung metastasis through different mechanisms. Our work may contribute to discover diagnostic markers and therapeutic targets.

Carcinoma, Adenoid Cystic ; secondary ; Carrier Proteins ; analysis ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; secondary ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; analysis ; Proteomics ; ras Proteins ; analysis

Sperm protein 17 (Sp17) is a testis-specific protein involved in acrosome reaction in spermatozoa. However, the Sp17 gene has been recently detected in normal non-testis tissues and malignant neoplasias. Therefore Sp17 may be a potential target for immunocontraception and a suitable target for tumor immunotherapy. This paper reviews the advances in the protein characterization, expression and distribution, and biological function of Sp17 and its clinical research.
Animals ; Antigens, Surface ; Carrier Proteins ; analysis ; physiology ; Contraception, Immunologic ; Humans ; Immunotherapy ; Male

BACKGROUND: Fascin is associated with motility in various transformed cells. Overexpression of fascin is known to aid in the progression of some cancers and is associated with a poor prognosis. E-cadherin is a major protein of epithelial cells and its expression is involved in the regulation of cell proliferation and differentiation. The aim of this study was to determine the expression pattern for fascin and E-cadherin and how it is related to the prognostic factors for renal cell carcinoma (RCC). METHODS: The expression of fascin and E-cadherin was evaluated in 208 RCCs including 175 clear cell, 20 papillary, and 9 chromophobe types using tissue array analysis. RESULTS: The expression of fascin increased as the tumor stage (p=0.00) and Fuhrman grade (p=0.00) increased. A high positive rate of expression for fascin was observed in cases with sarcomatoid changes (p=0.27). E-cadherin expression was seen in the distal tubules and collecting ducts of normal kidneys with a membranous pattern. The positive rate of expression for E-cadherin increased as the Fuhrman grade increased (1, 0%; 2, 23.2%; 3, 34.9%; and 4, 53.8%, p=0.00). An inverse correlation in RCCs was observed in the expression of fascin and E-cadherin (p=0.026, r=-0.158). CONCLUSIONS: In patients with RCC, the increased expression of fascin and E-cadherin was positively correlated to poor prognostic factors such as a higher Fuhrman nuclear grade and advanced pTNM stage.
Cadherins ; Carcinoma, Renal Cell ; Carrier Proteins ; Cell Proliferation ; Epithelial Cells ; Humans ; Kidney ; Microfilament Proteins ; Prognosis ; Tissue Array Analysis

OBJECTIVESTo detect the cholesterol ester transfer protein (CETP) levels in semen of infertile patients and evaluate the correlation between CETP in semial plasma and infertility.

METHODSOne hundred and sixty-three infertile patients and fifteen fertile males were selected randomly. The routine examination of ejaculates was fulfilled by computer aided semen analysis (CASA). The CETP levels in all seminal plasma samples and fifty-five serum samples were detected by ELISA method.

RESULTSThe CETP levels in infertile patients and fertile males were (2.21 +/- 1.23) microgram/L and (1.40 +/- 0.45) microgram/L, respectively. There were no significant differences between the two groups(P > 0.05). And there were no significant differences of CETP levels in seminal plasma among groups of azoospermia(n = 29), oligoasthenozoospermia (n = 58), oligospermia(n = 15), asthenozoospermia(n = 44) and normozoospermia(n = 17) in the infertile patients(P > 0.05). The CETP in seminal plasma and serum were detected in 55 infertile patients, and there was no correlation between CETP levels in seminal plasma and serum using Spearman analysis(r = 0.009, P > 0.05). The mean CETP level in seminal plasma was almost 1/1,000 of that in serum.

CONCLUSIONSThe CETP level in seminal plasma is extremely low and has no relation with the changes of sperm density or motility. It may ensure the integrity of sperm membrane before the sperm enters into female genital tract.

Adult ; Carrier Proteins ; analysis ; blood ; Cholesterol Ester Transfer Proteins ; Glycoproteins ; Humans ; Infertility, Male ; metabolism ; Male ; Middle Aged ; Semen ; chemistry

BACKGROUNDHuman urate anion exchanger (hURAT1) as a major urate transporter expressed on renal tubular epithelial cells regulates blood urate level by reabsorbing uric acid. Antibody is an important tool to study hURAT1. This study aimed, by genetic immunization, to produce mouse anti-hURAT1 polyclonal antibody with high throughput and high specificity and to detect the location of hURAT1 in human kidney.

METHODSHuman renal total RNA was isolated and the entire cDNA of hURAT1 was amplified by RT-PCR. The sequence of intracellular high antigenicity fragment (A280 to R349) was chosen by prediction software of protein antigenicity, and its cDNA was amplified from cDNA of hURAT1, and then cloned into pBQAP-TT vector to construct recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization. Mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-Flt3L, which helped to enhance the antibody's generation. After four weeks, the mice were sacrificed to obtain the anti-hURAT1 antibody from serum. The antibody was identified by western blot analysis and immunohistochemistry. At the same time, rabbit anti-hURAT1 antibody was produced by protein immunization. The specificity and efficiency between the rabbit and mouse anti-hURAT1 antibody were compared by western blot analysis and immunohistochemistry.

RESULTSThe entire cDNA of hURAT1 and cDNA of its intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization was confirmed by restriction digestion and sequencing. Both the mouse anti-hURAT1 antibody and rabbit anti-hURAT1 antibody recognized 58 kD hURAT1 and 64 kD glycosylated hURAT1 protein bands in western blot. Immunohistochemically, hURAT1 was located at the brush border membrane of renal proximal tubular cells. In addition, the throughput and specificity of the mouse anti-hURAT1 antibody were higher than those of the rabbit anti-hURAT1 antibody.

CONCLUSIONGenetic immunization can generate anti-hURAT1 polyclonal antibody of high throughput and specificity.

Animals ; Antibodies ; analysis ; Blotting, Western ; Carrier Proteins ; analysis ; immunology ; Female ; Humans ; Immunization ; Immunohistochemistry ; Kidney ; chemistry ; Male ; Mice ; Organic Anion Transporters ; analysis ; immunology ; Organic Cation Transport Proteins ; Plasmids ; Rabbits

OBJECTIVE: To explore the predictive value of heparin-binding protein (HBP) combined with sequential organ failure assessment (SOFA) score in patients with septic shock. METHODS: Seventy-eight patients with sepsis admitted to intensive care unit (ICU) of Henan Provincial People's Hospital from December 2016 to May 2017 were enrolled. Thirty healthy persons were enrolled as controls. The patient's gender, age, length of ICU stay, and blood culture results, white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), blood lactate (Lac), HBP, SOFA score, acute physiology and chronic health evaluation II (APACHE II) score, organ failure and vasoactive agents usage within 24 hours of admission were recorded. The differences in the above indicators between the groups were compared, and the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of HBP, SOFA score and their combination in patients with septic shock. RESULTS: All patients were enrolled in the final analysis, including 64 with sepsis and 14 with septic shock. Compared with the sepsis group, the proportion of patients with septic shock who were positive for blood culture, organ failure, and vasoactive agents was higher [57.1% (8/14) vs. 7.8% (5/64), 100.0% (14/14) vs. 65.6% (42/64), 100.0% (14/14) vs. 18.8% (12/64), all P < 0.01], SOFA and APACHE II scores were also higher (SOFA: 8.93±4.16 vs. 5.89±2.68, APACHE II: 22.29±4.89 vs. 15.28±5.14, both P < 0.01); however, there was no significant difference in gender, age or length of ICU stay between the two groups. Compared with the healthy control group, HBP, PCT, CRP and Lac levels were significantly increased in the sepsis group and the septic shock group. HBP in the septic shock group was significantly higher than that in the sepsis group (μg/L: 120.33±43.49 vs. 68.95±54.15, P < 0.05), but there was no significant difference in PCT, CRP or Lac between septic shock group and sepsis group [PCT (μg/L): 1.42 (0.47, 46.00) vs. 0.71 (0.19, 4.50), CRP (mg/L): 102.90±78.12 vs. 102.07±72.15, Lac (mmol/L): 1.81 (1.14, 3.65) vs. 1.59 (1.17, 2.24), all P > 0.05]. It was shown by ROC curve analysis that the area under the ROC curve (AUC) of SOFA score for predicting septic shock was 0.715 [95% confidence interval (95%CI) = 0.540-0.890, P = 0.012], and when the optimal cut-off value was 7.5, the sensitivity was 64.3%, the specificity was 76.6%. The AUC of HBP was 0.814 (95%CI = 0.714-0.913, P < 0.001), and when the optimal cut-off value was 89.43 μg/L, the sensitivity was 78.6%, the specificity was 76.6%; when the two were combined, the AUC was 0.829 (95%CI = 0.724-0.935, P < 0.001), the sensitivity was 92.9%, and the specificity was 61.9%. CONCLUSIONS HBP can be used as a biological indicator for predicting septic shock, and the accuracy of predicting septic shock can be improved with the combination of SOFA score.
Antimicrobial Cationic Peptides/analysis* ; Blood Proteins/analysis* ; Carrier Proteins/analysis* ; Female ; Humans ; Male ; Organ Dysfunction Scores ; Predictive Value of Tests ; Shock, Septic/diagnosis*

BACKGROUND/AIMS: In order to identify microsatellite instability (MSI), the test based on the polymerase chain reaction (PCR) can be used. However, PCR is not routinely performed in all hospital laboratories. Recently, immunohistochemistry (IHC) for MLH1 and MSH2 proteins has been reported as a rapid and useful method for MSI. However, the efficacy of IHC in the detection of the MSI has not been well established. The aim of this study was to evaluate the usefulness of IHC in the detection of the MSI by comparing it with the test results using PCR in colorectal cancer (CRC). METHODS: Paraffin-embedded normal and tumor tissues from seventy-five patients who underwent surgical resection of CRC were used. Abnormal expression of MLH1 and MSH2 protein was determined by IHC using MLH1 and MSH2 antibodies. Normal and tumor DNAs were obtained from thirty CRC tissues that showed abnormal expression of MLH1 and MSH2 proteins by IHC. The MSI status was confirmed by PCR using five markers. RESULTS: Thirty tumors showed abnormal expression of MLH1 and MSH2 proteins by IHC, but only three tumors out of them were confirmed to have MSI by PCR. CONCLUSIONS: This result suggests that IHC with MLH1 and MSH2 antibodies does not seem to be a useful method to identify MSI in CRC, therefore PCR is required for detection of the MSI.
Adaptor Proteins, Signal Transducing ; Aged ; Carrier Proteins ; Colorectal Neoplasms/*genetics ; DNA-Binding Proteins/*analysis ; Female ; Humans ; Immunohistochemistry ; Male ; *Microsatellite Repeats ; Middle Aged ; MutS Homolog 2 Protein ; Neoplasm Proteins/*analysis ; Nuclear Proteins ; Polymerase Chain Reaction ; Proto-Oncogene Proteins/*analysis

OBJECTIVE: The sensitivity of heart-type fatty acid binding-protein (H-FABP) in the postmortem diagnosis of myocardial ischemia was explored. METHODS: The changes of H-FABP staining in normal, infarcted and suspected ischemia of myocardial cells were studied by immunohistochemistry. RESULTS: There was no depletion in normal control group, and obvious depletion was observed in myocardial infarcted group. Among 9 suspected myocardial ischemia group, 3 cases showed obvious depletion and 3 cases showed vague depletion for H-FABP, there were obvious depletion of Mb in 4 suspected myocardial ischemia cases and vague depletion in 2 cases for Mb. It is indicated that H-FABP can be used to diagnose early myocardial ischemia. CONCLUSION H-FABP is quite sensitive and useful for the diagnosis of early myocardial ischemia.
Biomarkers/analysis* ; Carrier Proteins/analysis* ; Creatine Kinase/blood* ; Enzyme-Linked Immunosorbent Assay ; Fatty Acid-Binding Proteins ; Humans ; Immunohistochemistry ; Myocardial Ischemia/metabolism* ; Myocardium/pathology* ; Myoglobin/metabolism* ; Sensitivity and Specificity

Backgrond: Fascin is an actin-bundling protein that induces membrane protrusions and it increases cell motility in various transformed cells. Esophageal cancer is one of the most lethal malignancies, and it exhibits extensive local invasion or frequent regional lymph node metastasis even after curative surgery. We investigate the expression of fascin by performing immunohistochemistry to evaluate the clinical characteristics and prognostic significance of its expression in esophageal cancer patients. MATERIAL AND METHOD: Immunochemistry for fascin was performed on 76 tumor samples from 76 patients who underwent esophageal cancer operations. The expression levels of fascin in the 76 esophageal cancer tissues were compared with those in the corresponding normal esophageal epithelium. The fascin-positive samples were defined as those showing more than 75% of fascin-positive cells. RESULT: Overall, a fascin positive expression was detected in 39 (51.3%) out of the total 76 cases. The tumors with positive fascin expression tended to more frequently show a higher stage (p=0.030), and a higher T-factor (p=0.031). The prognosis of the fascin negative group was significantly better than that of the fascin positive group (p=0.004). Multivariate analysis revealed that lymphovascular invasion and the fascin expression were independent prognostic factors. CONCLUSION: Fascin was expressed in 51.3% of the esophageal cancer tissues, and a positive expression of fascin was associated with more advanced tumor progression and recurrence. Our study suggests that the fascin expression may be an independent prognostic factor for an unfavorable clinical course for those patients suffering with esophageal cancer.
Carrier Proteins ; Cell Movement ; Epithelium ; Esophageal Neoplasms ; Humans ; Immunochemistry ; Immunohistochemistry ; Lymph Nodes ; Membranes ; Microfilament Proteins ; Multivariate Analysis ; Neoplasm Metastasis ; Neoplasm Proteins ; Prognosis ; Recurrence ; Stress, Psychological

After collecting calcareous corpuscles from plerocercoid of Spirometra mansoni (sparganum), we evaluated the antigenic values of calcareous corpuscles binding proteins obtained from the cyst fluid of Taenia solium metacestodes. Immunoblot analysis revealed that cysticercosis patient sera strongly recognized 10 and 95 kDa calcareous corpuscles binding proteins. This result demonstrated that calcareous corpuscles are bound with major secretory antigenic proteins, which is possibly involved in the secretory pathways of the 10 and 95 kDa proteins presenting in the cyst fluid of T. solium metacestodes.
Animals ; Antigens, Helminth/*analysis ; Carrier Proteins/*immunology ; Cysticercosis/diagnosis/immunology ; Helminth Proteins/*immunology ; Humans ; Immunoblotting/methods ; Molecular Weight ; Serologic Tests ; Sparganum ; Taenia solium/*chemistry/immunology