Investigation of Immune-Regulatory Effects of Mageumsan Hot Spring via Protein Microarray In Vitro

BACKGROUND: Empirical evidences for efficacy of hot spring (HS) water in inflammatory skin disorders have not been substantiated with sufficient, immunological “hard evidence”. Mageumsan HS water, characterized by its weakly-alkaline properties and low total dissolved solids content, has been known to alleviate various immune-inflammatory skin diseases, including atopic dermatitis (AD). OBJECTIVE: The trial attempted to quantitatively analyze in vitro expression levels of chemical mediators in cutaneous inflammation from HaCaT cell line treated with Mageumsan HS, and suggest the likely mode of action through which it exerts the apparent anti-inflammatory effects in AD. METHODS: Using membrane-based human antibody array kit, customized to include 30 different, keratinocyte-derived mediator proteins, their expression levels (including interleukin [IL]-1, IL-6, IL-8, thymic stromal lymphopoietin, thymus and activation-regulated chemokine, and granulocyte macrophage colony-stimulating factor) were assessed in vitro. Selected key proteins were further quantified with enzyme-linked immunosorbent assay. RESULTS: There was a clear pattern of overall suppression of the mediators, especially those noted for their pro-inflammatory role in AD (monocyte chemoattractant protein [MCP]-1, regulated on activation, normal T cell expressed and secreted, cutaneous T-cell-attracting chemokine, Eotaxin, and macrophage inflammatory protein-1α, etc.). Also, reduced expression of involucrin and cytokeratin 1 was also reduced in the HS-treated group. CONCLUSION: The present study has shown that Mageumsan HS water may exert its effects on inflammatory skin disorders through regulation of proinflammatory cytokines. These evidences are to be supported with further future investigations to elucidate immunological mechanism behind these beneficial effects of HS water in the chronically inflamed skin of AD.

Identification of the Thioredoxin-Like 2 Autoantibody as a Specific Biomarker for Triple-Negative Breast Cancer / 한국유방암학회지

Triple-negative breast cancer (TNBC) has a higher risk of death within 5 years of being diagnosed than the other forms of breast cancer. It is the second leading cause of death due to cancer among women. Currently, however, no diagnostic blood-based biomarker exists to identify the early stages of TNBC. To address this point, we utilized a human protein microarray system to identify serum autoantibodies that showed different expression patterns between TNBC and normal serum samples, and identified five autoantibodies showing TNBC-specific expression. Among them, we selected the thioredoxin-like 2 (TXNL2) autoantibody and evaluated its diagnostic relevance by dot blot analysis with the recombinant TXNL2 protein. We demonstrated that the TXNL2 autoantibody showed 2- to 6-fold higher expression in TNBC samples than in normal samples suggesting that serum TXNL2 autoantibodies are potential biomarkers for TNBC.

Clinical value of tuberculosis protein chip in the diagnosis of tuberculosis infection / 中华检验医学杂志

Objective To investigate the Mycobacterium tuberculosis infection status and Clinical Characteristics in Yongchuan District, Chongqing by Tuberculosis Protein Chip.Methods Compared the conventional method to detect Mycobacterium tuberculosis in infectious department outpatient of Yongchuan Hospital , Chongqing Medical University from July 2014 to June 2015.Tuberculosis protein chip was selected to detect the Mycobacterium tuberculosis infection in Yongchuan area.Chi-square Test was applied to analyze the results.Results The positive rate of Tuberculosis Protein Chip, T-SPOT.BT, DNA Chip, Golden immnnochromatog-raphy, Acid-fast staining were 81.5%, 90.7%, 89.5%, 63.5% and 38.3%respectively on 162 cases of Pulmonary tuberculosis.The five methods were considered significant difference on the diagnosis of Pulmonary tuberculosis ( P0.05 ).The highest positive rates of anti-LAM was 94%.Conclusion The results of Tuberculosis protein chip are reliable on pulmonary tuberculosis and extrapulmonary tuberculosis diagnosis.

Relationship between plasma TRAIL before 20 weeks′gestation and pregnancy induced hypertension / 中华检验医学杂志

Objective To assess the relationship between maternal plasma tumor necrosis factor-related apoptosis-inducing ligand ( TRAIL ) before 20 weeks′gestation and hypertensive disorder complicating pregnancy (HDCP);and to evaluate the predictive value of plasma TRAIL for HDCP.Methods A 2-phase screening/validation study was designed.In the screening phase , a nested, case-controlled study was performed , the plasma samples collected before 20 weeks′gestation from 20 women who later developed HDCP and 20 age-and gestation week-matched controls were tested in prospective screening test for protein expression profiling during pregnancy and HDCP.Plasma samples were analyzed by a human protein microarray technology designed to detect 507 proteins simultaneously.Differently expressed proteins′functional annotation and clustering were performed by using of Database for Annotation , Visualization and Integrated Discovery ( DAVID) and Gene Ontology ( GO) database.The TRAIL level of plasma samples obtained before 20 weeks′gestation from 53 women who later developed HDCP and 106 similarly matched controls were further validated by ELISA and 62 clinical risk factors were investigated.Logistic regression and ROC analysis were used to evaluate the relationship between TRAIL and HDCP and its predictive value for HDCP.Results In protein microarray analysis , 23 proteins expressed differently before 20 weeks′gestation between the two groups.Further validation results showed that TRAIL levels in HDCP patients were lower significantly (45.7 ±13.1) pg/ml than those in healthy pregnant controls (51.2 ±14.7)pg/ml, P=0.021.Multiple factor logistic regression analysis of 159 pregnancies showed that three features were finally entering the logistic model, they were:anemia (OR=4.87, 95% CI 1.05-24.26), pre-pregnancy BMI (OR=1.72, 95% CI 1.35 -2.19) and TRAIL (OR=0.96, 95% CI 0.92 -0.99).The predictive accuracy of logistic model was 81.8%.The model significantly increases the predictive value (AUC=0.81, 95%CI 0.73-0.87) compared to TRAIL as independent predictor (AUC=0.59, 95%CI 0.51-0.67).Conclusions Totally 23 proteins were expressed differentially before 20 weeks′gestation in plasma of women who later developed HDCP , confirming that HDCP is a heterogeneous disease with different biological changes.The data suggests that plasma TRAIL levels relate with the development of HDCP and its combination with pre-pregnancy BMI and anemia have a high predictive value for HDCP before 20 weeks′gestation.

Analysis of differential expressed protein in the intestinal mucosa of patient with inflammatory bowel disease / 中华消化杂志

Objective To investigate differential expressed protein in the intestinal mucosa of patients with inflammatory bowel disease (IBD) with antibody chips,and to explore the possible role of the screened proteins in pathogenesis of IBD.Methods The mucosa tissues of nine patients with ulcerative colitis (UC),nine patients with Crohn's disease (CD) and nine control individuals were collected.After total protein of each group was extracted,the differential expressed protein of each group was analyzed by Raybiotech L-series human cytokine antibody chips.The mucose tissues of other nine patients with UC,nine patients with CD and nine control individuals were collected,and were used to verify the greatly differential expressed proteins by Western blot.The t-test was performed to compare two groups.Results Compared with the control group,there was significantly difference in 263 cytokines of UC group,and 414 cytokines of CD group.And then the higher expressions of herpes virus entry mediator,leukemia inhibitory factor and platelet factor 4 in the mucosa tissues of IBD patients were confirmed by Western blot and the differences were statistically significant (UC:t=23.85,9.53,18.88; CD:t=13.54,16.65,13.67,all P＜0.01).Conclusion The screened differential expressed cytokines in the mucosa tissues of IBD patients by cytokine antibody chips could be helpful to reveal the pathogenesis of IBD and discover new molecular biomarkers.

Multiple autoantibody detection in esophageal cancerous or precancerous serum using tumor-associated antigens microarray / 肿瘤

Objective: To identify serum autoantibody biomarkers by an tumor-associated antigens (TAAs)-based microarray in the patients with esophageal precancerous lesions (EPLs) and patients with esophageal cancer (EC). Methods: The antigens microarray with 55 TAAs was used to detect the serum autoantibodies of the residency-, age-, and sex-matched 26 pairs, which included 26 cases with EC, 26 cases with EPLs and 26 normal controls. The cut-off value was defined as mean ± 2 standard deviation of the expression levels of the antibodies in the normal control group. The value corresponding to the expression level of the antibody beyond the range of cut-off value was considered seropositive. Results: Five autoantibodies (CYFRA21-1, CA72-4, NY-ESO-1, GAGE-7 and SCCA) were significantly up-regulated, and others were down-regulated. Twenty-five autoantibodies had significant difference among EC, EPLs and normal controls which were found by multi-factor analysis of variance. Serum antibodies could be detected by 9 antigens, including CA72-4, CCNB1, CDKN2A, NY-ESO-1, CYFRA21-1, E2F1, ERBB2, GAGE-7 and SCCA. The positive detection rates of CYFRA21-1 were the highest in EC and EPLs, which were 61.54% and 60.00%, respectively. Furthermore, other antigens with higher seropositive rates were NY-ESO-1 (50.00% for EC, 52.00% for EPLs), SCCA (46.15% for EC, 48.00% for EPLs), GAGE-7 (46.15% for EC, 24.00% for EPLs) and CA72-4 (34.62% for EC, 16.00% for EPLs) in turn. The seropositive rates of these 9 antigens in combined detection panel were 88.46% and 84.00% for EC and EPLs, respectively. Conclusion: CYFRA21-1 is possibly used as an autoantibody indicator for early screening of EC and EPLs. The combined detection of the 9 antigens is likely to improve the sensitivity of autoantibody detection in EC and EPLs serum samples. Copyright © 2013 by TUMOR.

Detection and distribution of Helicobacter pylori antibodies by protein chip in gastroduodenal disease / 中国医师杂志

Objective To detect Helicobacter pylori (H.pylori) multiple antibodies of urease (Ure),cytotoxin associated gene A protein (CagA),vacuolating toxin A (VacA),heat shock protein 60 (Hap60) and nitroreductase ( RdxA),and disclose their relations with chronic gastritis,peptic ulcer and gastric cancer.Methods A volume (3 ml) of venous blood was taken from 300 patients of gastroduodenal disease diagnosed by endoscopy,to centrifuge and detect antibodies of Ure,CagA,VacA,Hsp60,RdxA by protein chip technique.Results The infective rates of H.pylori in chronic gastritis,peptic ulcer,and gastric cancer were 34.0％,58.0％,34.0％ ( P ＜ 0.01 ),respectively.In the H.pylori positive chronic gastritis,peptic ulcer and gastric cancer,the positive rates of CagA antibody were 54.9％,75.9％,64.7％ ( P =0.070) ; the positive rates of VacA antibody were 31.4％,22.4％,17.6％ ( P =0.412) ;and the positive rates of Hap60 antibody were 56.9％,48.3％,41.2％ ( P =0.466),respectively.The total positive rate of RdxA antibody was 4.0％ (5/126).Conclusions H.pylori infection and virulence factor CagA are closely related to peptic ulcer,while it did not show the exact correlation between VacA,Hsp60 and gastroduodenal disease.The level of RdxA antibody can not represent the level of H.pylori resistance to metronidazole.

Expression and clinical signifcance of COX-2 and telomerase in esophageal carcinoma and its resection margin / 肿瘤研究与临床

Objective To investigate the expression and clinical significance of COX-2 and telomerase activity in esophageal carcinoma tissues(ECT)and resection margin of the esophagus(RME),and to analyze their diagnostic and prognostic value for esophageal carcinoma.Methods The expression of COX-2 and telomerase activity in ECT and RME was examined in 82 patients with esophageal carcinoma and 40 normal esophageal epithelium tissues(NEET) by tissue array with EnVision imimunohistochemistry.Their correlations with the clinicopathologic factors were analyzed statistically.Results The positive expression rates of COX-2 and telomerase in ECT and RME were significantly higher than those in NEET(82.9 ％,29.3 ％vs 12.5 ％;and 87.8 ％,18.3 ％vs 5.0 ％;respectively;all P ＜ 0.05).The expression of COX-2 and telomerase in ECT and RME was correlated to TNM stage,tumor cell differentiation and lymph node metastasis (all P ＜0.01),while both expression in RME was closely related to anastomotic recurrence following resection of esophageal carcinoma (P＜0.01).The survival rate in esophageal carcinoma patients with the positive expression of COX-2 and telomerase in RME could be much lower than those with negative expression of COX-2 and telomerase in RME (P=0.000,Log-rank test).COX-2 expression was positively correlated to telomerase in ECT and RME (r=0.786,0.218,P＜0.05).Conclusion COX-2 and telomerase might be important biological markers for malignant transformation and invasion and metastasis of esophageal carcinoma.The activity of COX-2 and telomerase in RME could prognosticate anastomotic recurrence,and could be potential biomarkers evaluate the surgical treatment and prognosis of esophageal carcinoma.

Screening of serum proteomic patterns and tissue-specific genes for early liver metastasis of colorectal cancer / 第二军医大学学报

To screen for the serum proteomic patterns and related genes in early liver metastasis of colorectal cancer by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)and the RT2 Profiler TMPCR Array Human Tumor Metastasis (PAHS-028A) chip, so as to provide theoretical evidence for diagnosis of early liver metastasis of colorectal cancer. Methods The serum proteins of 20 colorectal cancer patients and 20 patients with early liver metastasis of colorectal cancer were detected by WCX 2 chip and SELDI-TOF-MS. PCR gene chips were used to screen the differentially expressed genes between the primary tumor and the liver metastases. Results SELDI-TOF-MS found that, when the M/Z values ranged 2000-30000, the contents of two proteins (3774 and 11851) were significant different in three samples. PCR gene chip found that the expressions of following genes were significantly higher in the primary colorectal cancer specimens than in the liver metastatic nodules: ACTB, APC, CTNNA1, NR4A3, MMP10, CTSL1, RB1, HPSE, ETV4, GNRH1, CDKN2A, KISS1R, IL8RB, ITGA7, ITGB3, DENR, RPSA, CXCR4, MYCL1, NME2, PNN, SMAD4, MMP11, SRC, RORB, SSTR2, SYK, TCF20, MMP3, TIMP2, TIMP3, TIMP4, and TRPM1;and the following genes were significantly higher in the liver metastases than in the primary tumors: MMP9, FN1, CST7, and CCL7. Conclusion SELDI-TOF-MS and gene chip technique can provide a theoretic basis for the mechanism, diagnosis and treatment of early liver metastasis of colorectal cancer.

HCC serum associated proteins screened by SELDI-TOF-MS analysis / 中华检验医学杂志

Objective To screen potential serum HCC associated proteins with low molecular weight and low abundance for better understanding the pathological mechanism of HCC and discovering new biomarkers.Methods All serum samples were collected from 81 HBV-related HCC patients,43 chronic hepatitis B patients and 36 cirrhosis patients.Serum protein fingerprint profiles were first generated by selected WCX2 protein chip integrating with SELDI-TOF-MS,and then normalized and aligned by Ciphergen SELDI Software 3.1.1 with Biomarker Wizard.Comparative analysis of the intensity of corresponding protein fingerprint peaks in normalized protein spectra was performed.Some protein peaks with significant difference among HCC,cirrhosis and chronic hepatitis B groups were found.The reproducibility of the SELDI system was assessed before serum protein fingerprint profiles analysis.Results The intra-and inter-assay CV for intensity and m/z in this SELDI system were 17.46% and 0.024%,and 17.74% and 0.024% respectively.Total 128 protein fingerprint peaks between 2 000 to 30 000 Da were identified under the condition of signal to noise＞5 and minimum threshold for cluster＞20%.Eighty-seven proteins were found to significantly expressed between HCC and cirrhosis groups(P＜0.05).Of the above differential proteins,forty-five proteins had changes greater than two fold,including 15 up-regulated proteins and 30 downregulated proteins in HCC sera.Between HCC and chronic hepatitis B groups,nine of fifty-two differential proteins(P＜0.05) had intensities of more than two folds,including 2 up-regulated proteins and 7 downregulated proteins in HCC sera.Between cirrhosis and chronic hepatitis B groups,twenty-eight of seventynine significantly differential proteins(P＜0.05) changed greater than two folds in intensity,including 17 up-regulated proteins in cirrhosis seru and 11 down-regulated proteins in chronic hepatitis B sera.Analysis of above leading differential proteins among three diseases using subtraction difference mode,the 5 common down-regulated proteins 2 870,3 941,2 688,3 165 and 5 483 m/z in HCC sera and 2 common up-regulated proteins 3 588 and 2 017 m/z in cirrhosis and HCC sera were screened.But no statistic difference in the level of protein 2 017m/z was found between HCC group and normal group inour previous study.Conclusion Because the interference of unspecific proteins from hepatitis B and cirrhosis could be eliminated partly in HCC sera through subtraction difference analysis,these 6 common differential proteins (2 870,3 941,2 688,3 165,5 483,3 588 m/z)have obvious advantages of increased specificity for evaluating the pathological state of HCC and might become promising candidate biomarkers in the diagnosis of HCC.

Screening serum markers of primary biliary cirrhosis with high-throughput protein chip encoded by the human genes / 中华检验医学杂志

Objective To screen serum markers in patients with PBC by high-throughput protein chips encoded by the human genome. Methods High-throughput protein chips (contains a total of 38 400protein spots, including 17 718 human genes encoding proteins) were used to screen sera from 21 PBC patients, 20 disease control patients and 10 normal controls. Bioinformatics software was used to analyze information and statistical software was used to analyze the data to confirm the serum markers of PBC. Results The detection rate of protein spots using anti-GST antibody on the chip was 97. 6%, and the signal intensity correlation coefficient of double protein spots was 0. 98. Four serum markers( PDHA1, DBT,DLAT and HK1 )were screened by high-throughput protein chips between PBC group and the control group with a statistically significant. The positive rate of the four markers in the three groups was 66. 67% ( 14/21), 5.00% (1/20) and 0(0/10); 57. 14% ( 12/21 ), 5.00% (1/20) and 0(0/10); 52. 38% ( 11/21 ),0(0/20) and 0(0/10); 52. 38% ( 11/21 ), 0(0/20) and 0(0/10) respectively. All the four markers were different in the three groups with statistically significant (PDHA1 :x2 = 16. 79, P ＜0. 01 ;Fisher exact test,P=0. 000; DBT:x2 =12.86, P＜0. 01;Fisher exact test, P=0. 004; DLAT and HK1:Fisher exact test,P ＜0. 01 or 0. 05). Of those markers, antibodies to PDHA1, DBT and DLAT were the component of AMAM2 which had been used as the marker of PBC. Antibody to HK1 was identified as new marker of PBC,whose sensitivity to PBC was 52. 38% and specificity was 100. 00%. There were no serum marker were screened between the AMA-M2 positive and negative PBC patients. Only antibody to CENPB was identified to be significantly expressed between the ACA positive and negative PBC patients ( Fisher exact test, P =0. 000). Conclusions High-throughput protein chip encoded by the human gene is a technology for quick and comprehensive screening of new markers of PBC. Antibodies to HK1 could be used as new marker for PBC with highly sensitivity and specificity. No serum marker is found between the AMA-M2 positive and negative PBC patients whereas only antibody to CENPB is identified as marker between the ACA positive and negative PBC patients.

Potential role of multiple tumor markers chip in preoperative diagnosis of gastric cancer / 国际肿瘤学杂志

Objective To explore the potential value of multiple tumor markers chip( C- 12) in preoperative diagnosis of gastric cancer. Methods The serum levels of 12 rumor markers were measured in 45 gastric cancer patients, 38 benign gastrosis patients and 65 normal controls by use of C-12 in order to find out the most levels of CA199, CEA, CA242, AFP and CA125 in the gastric cancer patients were significantly higher than those of the benign gastrosis patients and normal controls. Moreover, the serum levels of β- HCG and HGH were also significantly higher in gastric cancer group than benign gastric disease group and control group ( P ＜sis of gastric cancer. CEA is the TM with the highest sensitivity, validity and negative predictive value of 57.8% ,81.8% and 77.1% ,respectively whereas CA242 is the TM with the highest specificity and positive CEA + CA125 + CA199 and CEA + CA242 + CA199 + CA125, respectively. The sensitivity, specificity and validity of the best combination of 2 TMs, 3 TMs and 4TMs for gastric cancer were not statistically significantly different from those of C-12 and the best TM ( P ＞ 0.05 ). Conclusion The multiple tumor markers chip ( C-12 ) has a relatively high value in the preoperative diagnosis of gastric cancer. The best combinations of 2 TMs ( CEA + CA125) ,3 TMs ( CEA + CA125 + CA199 ) and 4TMs ( CEA + CA242 + CA199 + CA125 ) for gastric cancer diagnosis could be sufficient to replace the combination of 12 TMs.

Early diagnosis of diabetic nephropathy using protein pattern based on urinary biomarkers / 中华检验医学杂志

Objective To search for protein markers in urine from patients with diabetic nephropathy by proteomic method and discuss its clinical significance in laboratory diagnosis of diabetic nephropathy. Methods This study included 129 patients with diabetic nephropathy, 61 diabetes mellitus patients, and 102 healthy volunteers. The urinary protein profiles were obtained using surface-enhanced laser desorption-ionization time of flight mass spectrometry (SELDI-TOF-MS) and Au Chip (ProteinChip Gold Array). The differential peaks were screened by Biomaker Wizard software and the decision tree pattern was developed by Biomarker Patterns Software (BPS). The model was blindly tested to validate diagnostic efficiency. Some differentially expressed protein was preliminarily identified according to the molecular weight as compared with mass spectrometry data of standard proteins. Results Totally 40 distinguished protein peaks(t value: - 9.81-24.52, P < 0.05) were obtained after comparing the samples between diabetic nephropathy and the control groups. The peak with m/z 66 916 was automatically screened by BPS to develop decision tree pattern. The pattern was blindly tested and yielded a sensitivity of 98.7% (78/79) and a specificity of 98.2% (111/113). After we compared results from diabetic nephropathy with those from diabetes mellitus, twenty-four differential peaks were obtained in diabetic nephropathy (t value: -6.95-14.45,P < 0.05). The peaks with m/z 4 008, 11 619 and 66 916 were automatically screened by BPS to establish decision tree pattern. The model was blindly tested and yielded the sensitivity(129/129) and specificity(61/61) of 100%. After we compared our results with mass spectrometry data of standard proteins, the four differentially expressed proteins with m/z 11 619, 23 529, 66 916 and 79 378 were supposed to be β_2-microglobulin, α1-microglobulin, albumin and transfcrrin. Conclusion The preliminary results suggest that these SELDI-TOF and Au chip have the potential application value in identification of protein source and early diagnosis of diabetic nephropathy, and evaluation of renal injury.

Screening the serum proteomic patterns of Paget disease with SELDI-TOF-MS / 中国医师杂志

Objective To evaluate the diagnostic value of surface enhanced laser desorption/ionization time of flight mass spectrom-etry(SELDI-TOF-MS) for Paget disease. Methods The relative contents of serum proteins of 15 healthy people,15 patients with chronic eczema and 20 patients with Paget were detected by Weak cation exchanger protein chip (WCx2) and SELDI-TOF-MS. Results The con-tents of two proteins (3868Da,8876Da) had significant difference in healthy people and Paget disease patients(P<0.01). 291IDa, 3868Da and 5097Da protein peaks had significant difference in chronic eczema and Paget disease(P<0.05). Conclusion It shows great potentiality for early diagnosis and screening the tumor biomarkers of Paget disease with SELDI-TOF-MS.

Clinical value of multiple tumor marker protein chip detective system in diagnosis and detection of post-operative recurrence of breast cancer / 国际肿瘤学杂志

Objective To evaluate the clinical value of multiple tumor marker protein chip in diagno-sis and detection of postoperative recurrence of breast cancer.Methods The serum levels of 12 tumor makers (CA199,NSE,CEA, CA2A2,Ferritin,β-HCG,AFP,f-PSA,PSA,CA125,CA153 and HGH)were measured in 70 preoperative breast cancer patients, 32 recurrence patients,52 non-recurrence patients and 76 normal con-trois.Results ①The breast cancer group had significantly higher positive rate than that of the controls (P<0.05).The positive rates and serum levels of CA199,CEA,CA242,Ferritin,CAI25 and CA153 in breast cancer patients had those of control significant differences compared with groups (P<0.05).②The recurrence group had significantly higher positive rate than that of non-recurrence group (P<0.05).The positive rates and se-rum levels of CA199, CEA, Ferritin, CA125 and CA153 in the recurrence patients had significant differences compared with those of non-recurrence patients(P<0.05).③The positive rate of recurrence group had signif-icant difference compared with that of breast cancer group(P<0.05).Moreover,The positive rate and serum level of Ferritin in the recurrence patients had significant difference compared with that of breast cancer pa-tients.Conclusion The multiple tumor marker protein chip detective system has valid value of clinical appli-cation in the diagnosis and detection of postoperative recurrence of breast cancer.The combination detection of CA199, CEA, Ferritin ,CA125 and CA153 may be the economical and effective in the diagnosis and detection of postoperative recurrence of breast cancer.

Search of serum protein biomarkers for systematic lupus eryt.hematosus using protein chip tech- nology / 中华皮肤科杂志

Objective To study the changes of serum protein spectrum in patients with systematic lupus erythematosus (SLE) in order to screen specific protein markers. Methods Serum samples from 72 patients with SLE and 85 age- and sex-matched controls were assessed using surface-enhanced laser desorp-tion/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) with weak cation exchange (CM10) pro-rein chip. Forty samples from the patients and 50 control samples were randomly selected to serve as a pre-liminary training set; significantly different protein peaks were automatically chosen for the system training and development of a decision classification tree model. The validity of the model was then challenged with a blind test set (including another 32 samples from patients and 35 from human controls). Results A total of 73 effective protein peaks were detected within the mass/charge ratio (m/z) interval 2000 - 50000, among which, 15 protein peaks significantly differed between patients with SLE and controls (P < 0.01). Three pro-tein peaks with an m/z value of 4001, 6305 and 7356 were automatically chosen as a biomarker pattern in the training set that discriminated patients with SLE from controls with a sensitivity of 90.0% (36/40), speci-ficity of 92.0% (46/50) and accuracy rate of 91.1% (82/90). When the SELDI marker pattern was tested with the blinded test set, it yielded a sensitivity of 87.5% (28/32), specificity of 91.4% (32/35) and accuracy rate of 89.6% (60/67). Conclusions SELDI-TOF-MS protein chip could be used to screen serum protein for SLE, and the decision classification tree model with these biomarkers may favor the diagnosis of SLE.

Flow cytometric immnno-bead array assay for detection of platelet-specific autoantibodies / 中华检验医学杂志

Objective To establish a new flow cytometric immuno-bead array assay (FCIBA)to detect several platelet-specific autoantibodies simultaneously in a single serum sample.Methods A series of beads of different red fluorescent intensity were used for coating with different anti-platelet membrane protein monoclonal antibodies(anti-GP Ⅱb/a,anti-GP Ⅰ a/Ⅱ a,anti-GP Ⅳ,anti-GP Ⅰ b/Ⅸ and anti-HLAABC)to detect five platelet-specific autoantibodies in serum. The beads captured platelet antigenautoantiboay complex.Subsequently,different platelet-specific autoantibodies in a patient serum can be detected simultaneously by the flow cytometry.In addition,we evaluated the new FCIBA,and compared its results with modified antigen capture ELISA method(MACE).The new FCIBA was used to detect five platelet-specific autoantibodies in serums of autoimmune thrombocytopenic purpura(AITP)and nonautoimmune thromboeytopenic purpura patients.Results The new FCIBA can be used to detect five plateletspecific autoantibodies simultaneously(Anti-GP Ⅱ b/Ⅲ a,Anti-GP Ⅰ a/Ⅱ a,Anti-GP Ⅳ,Anti-GP Ⅰ b/Ⅸand Anti-HLA-ABC).The coefficient of variation(CV)of intra-repetition are 4.82%,6.09%,5.04%,5.73%and 5.30%,respectively.The dilution test results are in good logarithm linearity which are 0.997 2,0.996 6,0.998 8,0.996 5 and 0.998 2,respectively. The resuhs of the new FCIBA are highly correlated with those with MACE method,and the coefficient correlation were 0.928 9,0.922 4,0.889 4,0.910 0 and 0.913 4,respectively(P＜0.01).51.69%samples of AITP patients show positive for platelet-specific autoantibodies as detected by the new FCIBA.Among the AITP patients,the positivity of specific autoantibodies for anti-GPⅡb/ Ⅲ a,anti-GP Ⅰ a/Ⅱ a,anti-GP Ⅳ,anti-GP Ⅰb/Ⅸ and anti-HLA-ABC were 40.82%,24.45%,19.39%,32.65%and 17.35%.Among the 40 non-autoimmune thrombocytopenic purpura patients,none of platelet-specific autoantibodies in serum samples can be detected.Conclusion A new FCIBA is established successfully to detect five platelet-specific autoantibodies coefficient correlation.

Screening of mycosis fungo ides-associated serum proteins using SELDI protein chip technology / 中华皮肤科杂志

Objective To characterize mycosis fungoides (MF)-associated serum proteins by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS) technique. Methods Serum samples were collected following informed consent from 15 patients with MF and 15 patients with chronic eczema or neurodermatitis. Serum protein profiles were detected by CM10 protein chip combined with SELDI-TOF-MS. The proteins differentially expressed between MF and chronic eczema or neuroder matitis were assessed by the Zhejiang University Cancer Institute - ProteinChip Data Analysis System (ZUCIPDAS). Results A total of 329 protein peaks were detected from these patients, significant difference was observed in only 30 protein peaks. The optimal diagnostic model was developed by support vector machine algorithm with two protein peaks at Mass/Charge (m/z) 3939 and 5909 respectively. The expression of protein peak at m/z 3939 was higher in chronic eczema and neurodermatitis than in MF, while that at m/z 5909 appeared to express in an opposite way. In simulation training, this model proved to be able to distinguish MF from chronic eczema and neurodermatitis with both the specifity and sensitivity being 100 percent. The leave-one-out cross-validation also revealed a specificity and sensitivity of 100 percent for this model in the comparison of MF with chronic eczema and neurodermatitis. Conclusion These results suggest that SELDITOF-MS technique combined with bioinformatics is highly specific and sensitive in the diagnosis of MF.

Establishment of a diagnostic model from plasma proteomic patterns of ovarian serous cystadenocarcinoma patients using SELDI-TOF-MS technology / 肿瘤

Objective: To screen differentiated expressed proteins in plasma of ovarian serous cystadenocarcinoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) associated with bioinformatic support vector machines (SVM) and discuss how to establish algorithmic logical model for diagnosis of ovarian serous cys-tadenocarcinoma and its significance. Methods: SELDI-TOF-MS and CM10 chip were used to analyze the plasma samples from 26 ovarian serous cystadenocarcinoma women and 51 control women including 12 cases of ovarian cyst, 31 cases of uterous leiomyoma, 8 cases of ovarian benign cystadenoma. The data was analyzed by Biomarker Wizard software. The plasma proteomic diagnostic model for ovarian serous cys-tadenocarcinoma patients and control subjects were established by using SVM (a bioinformatic method). Results: Seventy-one differentiated protein peaks were screened by Biomarker Wizard software which were captured by SELDI-TOF-MS from CM10 chip (P <0.01). The proteomic profiling for ovarian serous cystadenocarcinoma was optimized by SVM re-screening. The key m/z value of these 7 proteins was 4 099, 4 477, 4 123, 4 081 and 3 938 (up-regulated), 8 785 and 13 783 (down-regulated). Three-fold cross validation followed by blinded determination demonstrated that the sensitivity and specificity of the established model were 84.62% and 96.08% separately, and the positive predictive value was 92.21% for differential diagnosis of ovarian serous cystadenocarcinoma patients. Conclusion: ProteinChip-mass spectrometry technology can rapidly and effectively screen differentiated proteins from the plasma of ovarian serous cystadenocarcinoma patients. Combined with SVM, a diagnostic model was generated from proteomic patterns of ovarian serous cystadenocarcinoma, which had potential significance for establishing diagnostic methods for ovarian cancer.

Microarray analysis of the relationship between PTEN and prognostic factors in primary breast cancer tissues / 肿瘤

Objective: To construct tissue microarray of breast carcinoma and to investigate the association of hepatocyte-specific phosphatase and tensin homolog (PTEN) protein expression with prognostic factors such as estrogen receptor (ER), progesterone receptor (PR), c-erb B-2, and Ki-67 in primary breast carcinoma. Methods: Tissue microarray of breast carcinoma was constructed. Immunohistochemical staining was used to detect the expression of PTEN, ER, PR, c-erb B-2, and Ki-67 inbreast carcinoma tissues. Results: PTEN was highly expressed in 66 out of 163 breast carcinoma tissues (40.5%). The positive rate of ER, PR, c-erb B-2, and Ki-67 was 51.5%, 35.6%, 34.4%, and 44.8%, respectively. The positive expression level of MN decreased with increase in auxiliary lymph nodes metastasis, histological grading, and TNM staging. They had negative correlations (P < 0.05). The positive expression level of PTEN had no evident correlation with tumor size and onset age. PTEN had positive correlation with the expressions of ER and PR protein (P < 0.01), and had negative correlation with the expressions of c-erb B -2 and Ki-67. Conclusion: There exists some relationships between low or loss of PTEN expression and the imbalanced expression of ER and PR and the growth and prognosis of breast carcinoma. PTEN may be considered as a potential indicator to judge the malignant degree of breast carcinoma. Combined detection of PTEN, ER, PR, c-erb B-2 and Ki-67 have significant importance and practical value for adjuvant therapy of breast carcinoma.