Profiling of differentially expressed proteins between fresh and frozen-thawed Duroc boar semen using ProteinChip CM10.

Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

Screening of serum protein biomarkers in hemorrhagic cerebral infarction by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technology.

BACKGROUND: Serum samples of patients with hemorrhagic cerebral infarction (HCI), cerebral infarction (CI), and healthy controls (HCs) were used to screen statistically different protein peaks as potential biomarkers and to establish a decision tree classification model. METHODS: The serum samples from clinically confirmed patients with HCI and CI from November 2018 to October 2019 were collected, along with those of HCs who visited our hospital during the same period. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) with CM10 ProteinChip was used to analyze the differences in serum protein expression profiles of 30 patients with HCI, 32 patients with CI, and 31 HCs in the training group, and a decision tree classification model was established. At the same time, the blind test group (18 patients with HCI, 21 patients with CI, and 17 HCs) was tested by a blind method. RESULTS: Model 1 was successfully established by software analysis with a mass-to-charge ratio of 3,495.2, 8,941.0, and 15,890.4 as a differential protein peak. The sensitivity, specificity, and accuracy of model 1 in distinguishing HCI from HCs were 86.8%, 87.1%, and 86.9%, respectively. After verification of model 1 by the blind test group, the results showed that the sensitivity, specificity, and accuracy were 88.9%, 94.1%, and 91.4%, respectively. The sensitivity, specificity, and accuracy of model 2 with a mass-to-charge ratio of 2,941.3 as a differential protein peak were 86.7%, 75.0%, and 80.6%, respectively. After verification of model 2 by the blind test group, the results showed that the sensitivity, specificity, and accuracy were 83.3%, 90.4%, and 87.2%, respectively. CONCLUSIONS: Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and CM10 ProteinChip can be used to screen serum protein markers in patients with HCI. Mass-to-charge ratio of 3,495.2, 8,941.0, 15,890.4, and 2,941.3 may be potential protein biomarkers of HCI and used to distinguish HCI patients from HCs and CI.

Anaphylatoxin C3a: A potential biomarker for esophageal cancer diagnosis.

Esophageal carcinoma is a common malignancy worldwide, with a low 5-year survival rate. As the majority of cases are diagnosed at an advanced stage, there is an urgent need for an effective biomarker for early diagnosis of esophageal cancer patients. Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was applied to detect the serum protein expression in esophageal cancer patients using ProteinChip software, and the results were analyzed and screened using Biomarker Patterns and SPSS16.0 software. The ELISA method was conducted to determine the concentration of anaphylatoxin C3a, which is one of the complement proteins, in the serum of esophageal cancer patients and non-esophageal cancer participants. A total of 144 effective differential expression protein peaks in the window of 1-10 kDa were obtained (P<0.05). M/Z 8,926.478 (P<10-6) protein peak was employed as the diagnostic biomarker for esophageal carcinoma. This established diagnostic biomarker has a sensitivity of 95% (19/20) and an accuracy of 100% (19/19) for positive prediction. The results suggested that anaphylatoxin C3a may be a promising biomarker in the diagnosis of esophageal carcinoma, and may play a key role in promoting esophageal carcinogenesis.

An efficient biomarker panel for diagnosis of breast cancer using surface-enhanced laser desorption ionization time-of-flight mass spectrometry.

Breast cancer (BC) is the most frequently diagnosed cancer that affects women worldwide. Early detection of BC is important to improve survival rates and decrease mortality. The aim of the present study was to investigate serum biomarkers using surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) to distinguish patients with BC from the healthy population and patients with benign breast diseases (BBDs). A total of 62 patients with invasive ductal carcinoma, as confirmed by histopathology, and 47 non-cancerous individuals (NCIs) [16 healthy controls (HCs) and 31 patients with BBD] were enrolled in the present study. Serum protein profiles were determined by SELDI-TOF-MS using an immobilized metal affinity capture array. Serum from patients with BC were compared with that from the HC group using univariate and multivariate statistical analyses. A total of 118 clusters were generated from the individual serum. Univariate analysis revealed that 5 peaks were significantly downregulated (m/z 1,452, 2,670, 3,972, 5,354 and 5,523; P<0.001) and 4 were upregulated (m/z 6,850, 7,926, 8,115 and 8,143; P<0.001) in patients with BC compared with the HC group. A comparison of patients with BC and patients with BBD revealed an additional 9 protein peaks. Among these, 3 peaks (m/z 3,972, 5,336 and 11,185) were significantly downregulated and 6 peaks (m/z 4,062, 4,071, 4,609, 6,850, 8,115 and 8,133) were significantly upregulated. A total of 3 peaks [mass-to-change ratio (m/z) 3,972, 6,850 and 8,115 (BC2)] were common in both sets. The results of the present study suggest that a 4 protein peak set [m/z 3,972, 6,850 and 8,115 (BC2) and 8,949 (BC3)] could be used to distinguish patients with BC from NCI.

Application of protein typing in molecular epidemiological investigation of nosocomial infection outbreak of aminoglycoside-resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosan has emerged as an important pathogen elated to serious infections and nosocomial outbreaks worldwide. This study was conducted to understand the prevalence of aminoglycoside (AMG)-resistant P. aeruginosa in our hospital and to provide a scientific basis for control measures against nosocomial infections. Eighty-two strains of P. aeruginosa were isolated from clinical departments and divided into AMG-resistant strains and AMG-sensitive strains based on susceptibility test results. AMG-resistant strains were typed by drug resistance gene typing (DRGT) and protein typing. Five kinds of aminoglycoside-modifying enzyme (AME) genes were detected in the AMG-resistant group. AMG-resistant P. aeruginosa strains were classified into three types and six subtypes by DRGT. Four protein peaks, namely, 9900.02, 7600.04, 9101.25 and 10,372.87 Da, were significantly and differentially expressed between the two groups. AMG-resistant P. aeruginosa strains were also categorised into three types and six subtypes at the distance level of 10 by protein typing. AMG-resistant P. aeruginosa was cloned spread in our hospital; the timely implementation of nosocomial infection prevention and control strategies were needed in preventing outbreaks and epidemic of AMG-resistant P. aeruginosa. SELDI-TOF MS technology can be used for bacterial typing, which provides a new method of clinical epidemiological survey and nosocomial infection control.

Serum proteomic signatures as biomarkers of primary biliary cirrhosis diagnosis and prognosis.

Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease considered as an autoimmune disease. To identify new biomarkers of PBC, serum profiling analysis using Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was employed. Twelve patients with either asymptomatic PBC (group 1, n=6) or PBC with a poor response to UDCA (group 2, n=6), were compared to healthy controls (group 3, n=6). Analysing the 18 sera by using four SELDI-TOF arrays under various conditions, we found four biomarkers of PBC at 5.9, 8.6, 8.9 and 9.0 kDa. The combination of the two arrays IMAC-40/Zn2+ and CM-10/pH 7 improved the positive diagnosis of this disease. We also found a biomarker of severity of PBC at 95.2 kDa on LSAX-30 array which characterized patients with a bad prognosis. In conclusion, our study identified several serum proteomics signatures as potential biomarkers of PBC for its diagnosis or prognosis.

Rapid detection of wild-type and mutated transthyretins.

BACKGROUND: Familial amyloid polyneuropathy is caused by a variant transthyretin, which is a serum protein secreted by the liver. We previously reported that mutated transthyretins were detected in serum samples by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). The aim of this study was to evaluate the clinical usefulness of SELDI-TOF MS for diagnosis of transthyretin-related amyloidosis. METHODS: We used 106 serum samples obtained from patients who were clinically suspected of having amyloidosis between February 2011 and April 2014. SELDI-TOF MS allowed analysis for transthyretin via a 3-h one-step procedure. RESULTS: Of the 106 patients, 51 are transthyretin amyloidosis. Mutated transthyretins were detected in serum samples from 30 of 51 patients with transthyretin amyloidosis. The results of genetic analysis showed that all of those patients had mutations in the transthyretin gene. For all 18 patients with senile systemic amyloidosis of 51 patients with transthyretin amyloidosis, SELDI-TOF MS detected only wild-type transthyretin peaks, not mutated transthyretin peaks. CONCLUSION: SELDI-TOF MS is a clinically useful tool for diagnosis of transthyretin-related amyloidosis.

The research on affinity of hospital separation carbapenems-resistant Acinetobacter / 国际检验医学杂志

Objective To study the protein fingerprinting of carbapenems resistant and susceptible Acinetobacter baumannii (Ab) strains ,investigate the clinical value of affinity distance in carbapenems resistant strains .Methods A total of 22 carbapenems resistant Ab strains and 18 carbapenems susceptible Ab strains were collected ,and bacterial protein fingerprinting was detected by surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) ,differentially expressed proteins was analyzed by Biomarker Wizard 3 .1 .Cluster analysis of differential expressed proteins was conducted on SPSS 19 .0 .Results The protein expression pattern of carbapenems resistant and sensitive Ab strains had significant difference (P< 0 .05) .Cluster anal-ysis showed carbapenems resistance Ab was given priority to with type A ,followed by type B .Conclusion The results of cluster a-nalysis carbapenems resistance of Ab protein fingerprinting could determine the distance of affinity relationship of Ab .It could pro-vide a theoretical basis for the infection and clinical epidemiology of Ab .

Application of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technology in evaluating anxiety in patients with malignant tumors / 肿瘤研究与临床

Objective To study the occurrence of anxiety in patients with malignant tumor, and in further explore the scientificity and clinical application value of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI) technology in anxiety detection. Methods 1 000 patients with malignant tumor were selected as research objects, the self-rating anxiety scale (SAS) was used to evaluate the anxiety on the first day of admission, the fasting venous blood of patients was taken on the next day morning. The SAS score and SELDI result were compared and analyzed. Results The effective sample number was 988. The SAS score was 56.32± 9.665, there were 454 cases of anxiety, the incidence rate of anxiety was 45.95 %. 470 cases were SELDI test positive, and the incidence rate of anxiety was 47.57 %. SAS was used as the gold standard to judge anxiety, the sensitivity and specificity of SELDI technology were 93.17 % and 91.20 %, the positive predictive value and negative predictive value were 90.00 % and 94.02 %, the total coincidence rate was 92.11 %. SAS score highly correlated with the abundance of SELDI, the fitting curve showed an up trend, and the correlation coefficient was 0.837. Conclusions The incidence of malignant tumor patients ' anxiety is relatively high. SELDI technique shows high sensitivity, specificity, total coincidence rate and correlation in anxiety detection, it can be used as an objective evaluation of anxiety.

Surface-enhanced laser desorption ionization time-of-flight mass spectrometry used to screen serum diagnostic markers of colon cancer recurrence in situ following surgery.

The aim of the present study was to identify specific serum biomarkers in patients with colon cancer recurrence in situ following surgery. The study was conducted at the Renmin Hospital of Wuhan University (Wuhan, China) between January 2012 and January 2014. Surface-enhanced laser desorption ionization time-of-flight mass spectrometry was used to compare and analyze the serum protein profiles of patients with (n=50) and patients without (n=50) recurrence in situ. Biomarker Wizard software was used to analyze and obtain the protein spectrum. In total, nine protein peaks demonstrated statistically significant differences between the recurrence and non-recurrence group (P<0.05), which included two protein peaks (7,731.3 Da and 8,266.5 Da). The two protein peaks were highly expressed in patients with colon cancer recurrence in situ following surgery, but lowly expressed in patients without recurrence. Therefore, the two protein peaks may represent potential biomarkers for the prediction of colon cancer recurrence in situ following surgical treatment.

Serum Protein Pattern Could Predict the Therapeutic Effect of First-Line Pemetrexed/Cisplatin Chemotherapy in Patients With Lung Adenocarcinoma.

BACKGROUND: In patients with advanced non-squamous non-small cell lung cancer (NSCLC), a pemetrexed/cisplatin (PP) regimen is considered as one of the preferred first-line treatments. However, only about half of the treated patients respond, and there is no clinically useful marker that can predict the response to the regimen. METHODS: We established a potential pattern for the prediction of efficacy of first-line PP chemotherapy in patients with lung adenocarcinoma, by using artificial neural networks (ANNs) analysis of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) in this preliminary study. RESULTS: The samples were randomly divided into training set and test set. From the test set, through cross-validation, the established protein pattern for PP separated the responders from the non-responders with a sensitivity of 95.8% and a specificity of 90.0%. CONCLUSION: It could be helpful for oncologists to select patients who could benefit from PP chemotherapy.

Clinical study of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technology in detecting the anxiety of patients with breast cancer / 肿瘤研究与临床

Objective To investigate the incidence of anxiety in patients with breast cancer,and further explore the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI) technology' s clinical value in the diagnosis of anxiety.Methods 121 breast cancer patients were selected as research objects by using convenience sampling method.Self-rating anxiety scale (SAS) was used to evaluate the anxiety on the first day of admission and SELDI was used on the next day to detect the serum from patients' fasting venous blood sample got in the morning after their consent,then anxiety-related protein fingerprints spectrums were selected.The SAS score and SELDI result were finally compared and analyzed.Results SAS score in 121 patients was 53.45±9.78,anxiety occurred in 63 cases (52.07 ％).Established the diagnostic model between 15 000+H and 16 800+H protein fingerprints abundance ≥5 ％,that was to say,abundance ≥5 ％ was judged as positive,otherwise negative.On this basis,patients can accurately be distinguished between anxious group and non-anxious group.Correct rate (total coincidence rate) was 91.74 ％,specificity and sensitivity respectively were 89.66 ％ (52/58) and 93.65 ％ (59/63),the positive predictive value was 90.77 ％ (59/65),and negative predictive value was 92.86 ％ (52/56).Conclusions Breast cancer patients have a high incidence of anxiety.SELDI technology has showed high sensitivity and specificity in anxiety detecting.As an objective assessment tool,it could have better prospects for clinical use.

Detection of pancreatic cancer with normal carbohydrate antigen 19-9 using protein chip technology.

AIM: To develop a method to differentiate pancreatic cancer patients from healthy or benign individuals when carbohydrate antigen (CA) 19-9 is normal. METHODS: Forty-one serum samples from patients with pancreatic lesions and blood samples from 20 healthy individuals were collected at the first stage of the experiment according to the enrolment criteria. General characteristics and some clinical features were carefully compared to ensure that the results were reasonable. All the blood samples were analyzed by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) combined with CM10 chips and a related bioinformatics analysis program to generate diagnostic models with different proteins. Forty-seven consecutive samples were tested at the next stage to verify the veracity and efficiency of the models. RESULTS: The sex, age, and serum CA19-9 levels among the three groups (malignant, benign, and healthy) were statistically matched (P values were 0.957, 0.145, and 0.382, respectively). Two patterns were generated. Pattern 1 with four proteins theoretically had a specificity and sensitivity of 100% in distinguishing pancreatic cancer from healthy individuals, while it was 86.7% and 86.4%, respectively, in the subsequent practical verification. The positive predictive value (PPV) of the model was 86.4%. One of the four proteins was expressed highly in pancreatic cancer while the other three were expressed weakly. Pattern 2 consisted of six proteins that showed a specificity of 70.0% and sensitivity of 77.3% for differentiating malignancy from benign tumors. Its PPV reached 85.0%. Only one of these six proteins showed high expression in the malignant group. CONCLUSION: SELDI-TOF-MS may facilitate diagnosis or differential diagnosis of pancreatic cancer when CA19-9 is normal. Pattern 1 may serve as a useful screening tool.

S100A proteins as molecular targets in the ocular surface inflammatory diseases.

The S100 proteins are calcium-binding proteins that are exclusively expressed in vertebrates, where they interact with enzymes, cytoskeletal proteins, receptors, transcription factors, and nucleic acids to regulate proliferation, differentiation, apoptosis, inflammation, cell migration, energy metabolism, and Ca(2+) homeostasis. In this review, we focus on the S100A8 and S100A9 members of the family that are involved in the regulation of neutrophil chemotaxis and inflammation related to ocular surface diseases such as dry eye, meibomian gland dysfunction, pterygium, and corneal neovascularization. In our previous studies, we have found that the levels of S100A8 and S100A9 were elevated in these inflammatory ocular diseases. For instance, S100A8 and A9 were found to be upregulated in pterygium tissues at both transcript and protein levels. These findings are consistent with the role of S100A8 and S100A9 proteins in activating the innate immune system in the eye via Toll-like receptors (TLRs) and altering the immune tolerance of the eye-associated lymphoid system. Recently, use of S100A8-targeting antibody has shown promising results in targeting corneal neovascularization. Injection of S100A8 has been shown to inhibit eosinophilic infiltration and thus may have potential therapeutic implications in allergic diseases.

Expression and significance of four kinds of serum protein/polypeptide in tuberculosis patients and healthy controls / 实用医学杂志

Objective To assess the efficacy of using four kinds of proteins / peptides to distinguish the tuberculosis patients from healthy people. Methods A, B, C and D were used to represent four proteins /peptides with 1 060, 1 944, 2 081 and 3 954 of mass to charge ratio (m / z) in serum, respectively. Levels A, B, C and D in serum of 57 patients with tuberculosis and 30 healthy people were determined by using the surface-enhanced laser desorption-ionization time of flight mass spectrometry (SELDI-TOF-MS). Then the differences of levels of f A, B, C and D were anlyzed between tuberculosis patients and healthy people. The efficacy of distinguishing tuberculosis patient from healthy people were evaluated by using diagnostic test evaluation method. Results (1) The levels of A, B, C and D were 1 ± 11, 1 597 ± 3 102, 460 ± 765 and 1 208 ± 1 003 in tuberculosis patients, while they were 123 ± 201, 47 ± 98, 36 ± 93 and 397 ± 355 in healthy people. (2) The area under the receiver operator characteristic (ROC) curve was 0.644, 0.848, 0.735 and 0.810 respectively. The serum levels of A, B, C and D could be used to distinguish tuberculosis patient from healthy people and the cut-off values of A, B, C and D were ≤166, ≥318, ≥48 and ≥728, respectively. Conclusions B, C and D have better performances to distinguish tuberculosis patients from healthy people , which may be regarded as new promising candidate markers for diagnosis of tuberculosis.

Time-of-flight mass spectrometry in screening serum-specific protein in patients with scrotal Paget's disease.

The weak cationic chip (WCX2) and the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) were used to test the serum differential proteins in 20 healthy persons, 20 patients with chronic scrotal eczema, and 30 patients with scrotal Paget's disease and test the specificity and sensitivity of screening scrotal Paget's disease with differential proteins. We found that the differences between the 5 protein peaks of the normal group and the scrotal Paget's disease group in the range of 2000 to 30 000 Da were statistically significant (P < .01) and the difference of 3 protein peaks between the scrotal eczema group and the scrotal Paget's disease group in the range of 2000 to 30 000 Da was statistically significant (P < .05). SELDI-TOS-MS technique has certain application value in the early diagnosis of the scrotal Paget's disease and screening for the specific tumor markers.

Application of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry technology for the diagnosis of colorectal adenoma.

The aim of the present study was to identify a specific biological marker for the diagnosis of colorectal adenomas through the analysis of variations in serum protein profiling in colorectal adenoma patients. The study was conducted at the Renmin Hospital of Wuhan University (Wuhan, China) between September 2011 and May 2012. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was performed to compare the serum protein profiles of 50 patients with colorectal adenoma and 50 healthy individuals. The obtained protein profiles were analyzed using Biomarker Wizard software. Twenty protein peaks were identified to exhibit differences in average intensity between colorectal adenomas compared with normal controls, including peaks 8,565.84, 8,694.51 and 5,910.50 Da, in which the intensity between the patients and control individuals was significantly different. Two peaks, 8,565.84 and 8,694.51 Da, were observed to be highly expressed in the colorectal adenomas, however, expression was low in the control samples. By contrast, 5,910.50 Da expression was low in the colorectal adenomas and high in the controls. The results of the current study indicate that the three protein peaks may represent specific biomarkers for colorectal adenomas.

The consumption of seaweed as a protective factor in the etiology of breast cancer: proof of principle.

Daily consumption of seaweed has been proposed as a factor in explaining lower postmenopausal breast cancer (BC) incidence and mortality rates in Japan. This clinical trial assessed the impact of introducing seaweed- to non-seaweed-consuming American postmenopausal women. Fifteen healthy postmenopausal women were recruited for a 3-month single-blinded placebo controlled clinical trial; five had no history of BC (controls) and ten were BC survivors. Participants ingested ten capsules daily (5 g day-1) of placebo for 4 weeks, seaweed (Undaria) for 4 weeks, then placebo for another 4 weeks. Blood and urine samples were collected after each treatment period. Urinary human urokinase-type plasminogen activator receptor concentrations (uPAR) were analyzed by ELISA, and urine and serum were analyzed for protein expression using surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry (SELDI-TOF-MS). Urinary creatinine standardized uPAR (in pg mL µg-1 creatinine) changed significantly between groups, decreasing by about half following seaweed supplementation (placebo 1, 1.5 (95 % CI, 0.9-2.1) and seaweed, 0.9 (95 % CI, 0.6-1.1) while placebo 2 returned to pre-seaweed concentration (1.7 (95 % CI, 1.2-2.2); p = 0.01, ANOVA). One SELDI-TOF-MS-identified urinary protein (m/z 9,776) showed a similar reversible decrease with seaweed and is reported to be associated with cell attachment. One serum protein (m/z 8,928) reversibly increased with seaweed and may be the immunostimulatory complement activation C3a des-arginine. uPAR is higher among postmenopausal women generally, and for BC patients, it is associated with unfavorable BC prognosis. By lowering uPAR, dietary seaweed may help explain lower BC incidence and mortality among postmenopausal women in Japan.

Proteomic profiling of heterotopic heart-transplanted rats using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry: potential biomarkers and drug targets.

OBJECTIVE: Methotrexate and rapamycin demonstrate an additive effect in prolonging cardiac allograft survival in a major histocompatibility complex mismatched rat model. The present study aimed to identify functional proteins involved in the allograft-protective effects of these two agents and reveal potential diagnostic markers for treating rejection. METHODS: Serum samples from heterotopic heart-transplanted LEW(RT-1(1)) rats (either without immunosuppressive treatment or treated with methotrexate alone, rapamycin alone, or methotrexate and rapamycin combined) were analysed by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Protein profiles obtained using a weak cation exchange ProteinChip® CM-10 array were then analysed using ProteinChip® Software. RESULTS: Of 28 rejection-related proteins identified, isoelectric point and mass information from two potential candidate proteins matched information from the UniProtKB/Swiss-prot database, suggesting them to be complement component C3f fragment and complement component 4A (C4A, anaphylatoxin). CONCLUSIONS: Proteomic analysis revealed 28 proteins as potential diagnostic markers of tissue rejection. Of these, 11 proteins may represent targets relating to the additive effects of methotrexate and rapamycin. Two protein peaks, with mass-to-charge ratios of 1950 Da and 8577 Da, may have potential for use in post-transplant diagnosis of rejection.

Application of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry in screening breast cancer markers / 肿瘤研究与临床

Objective To explore the application of serum SELDI proteomic patterns to screen breast cancer biomarkers.Methods Serum protein profiles of 110 breast cancer patients and 100 healthy controls were analyzed with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOFMS).The spectra were generated on weak cation exchange (WCX2) chips and protein peaks clustering and classification analyses were made using Biomaker Wizard software.Differences in protein intensity between breast cancer cases and controls were measured with the Mann-Whitney U test and adjusted for confounding in a multivariate logistic regression model.Results Forty-nine of these proteins were found to have statistically differential expression levels between breast cancer and normal control sera (P ＜ 0.05).Based on literatures reported,six protein biomarkers,with mass-to-charge ratio (M/Z) (4376,8126,8924,3264,3968,and 9180) were selected.Proteins with M/Z 4376,4126,and 8924 were statistically significantly decreased in breast cancer cases compared to those in healthy controls (P ＜ 0.05).Proteins with M/Z 3264,3968,and 9180 were significantly increased in breast cancer cases compared to those in healthy controls,Protein with M/Z 9180 was associated with TNM stage and Her-2 expression in breast cancer (P ＜ 0.05).Protein with M/Z 8926 was related with lymph node metastasis (P ＜0.05).Conclusion These results suggest that serum SELDI protein profiling can distinguish breast cancer patients from normal subjects with relatively high sensitivity and specificity.SELDI-TOF-MS plays a valuable role in the diagnosis of breast cancer and the discovery of new tumor-specific protein biomarkers.