Effects of clinical isolates of Streptococcus pneumoniae on THP-1 human monocytic cells.

Twenty­three clinical Streptococcus pneumoniae (SP) strains were isolated from blood and sputum specimens from the Second Affiliated Hospital of Wenzhou Medical College in 2009. These strains and the ATCC 49619 standard strain were cultured and suspended in normal saline (at a turbidity of 1.0 McFarland). The production of interleukin (IL)­8, intracellular adhesion molecule­1 (ICAM­1) and IL­10 in THP­1 cells following stimulation with the SP suspension was analyzed by an enzyme-linked immunosorbent assay. The concentrations of IL­8, ICAM­1 and IL­10 from the THP­1 monocytes were greater than those of the blank control following stimulation with the SP suspension. No significant difference was identified in the levels of IL­8, ICAM­1 and IL­10 secretion between THP­1 monocytes stimulated by blood­borne SP (bb­SP) and sputum­borne SP (sb­SP).

Roles of virulence genes (PsaA and CpsA) on the invasion of Streptococcus pneumoniae into blood system.

BACKGROUND: Streptococcus pneumoniae (SP) is the major cause of childhood mortality worldwide, we need to understand virulence genes of SP so can better target the treatment.We investigated the expression of virulence genes PsaA and CpsA in different strains of SP interacting with monocyte cell line (THP-1) or pneumocyte cell line (A549) and the possible mechanism of SP invasion of the blood system. METHODS: A total of 23 strains of SP were collected from hospitalized patients (blood-derived and sputum-derived) in the Second Affiliated Hospital of Wenzhou Medical College. The strains and ATCC 49619 were cultured, and RNAs were extracted. THP-1 and A549 cells were stimulated by different SP and ATCC 49619 for 4 h and 8 h, respectively. Quantitative real-time PCR was used to analyze the mRNA expression of PsaA and CpsA. The data were analyzed by SPSS 17.0. RESULTS: The mRNA level of PsaA and CpsA were all significantly increased in clinical SP strains when compared to ATCC49619 after tedTHP-1 and A549 cells were stimulated. Clinical SPs showed higher virulence compared with ATCC49619. CONCLUSIONS: The expression of CpsA is the basis of the pathogenicity of SP. The expression of virulence gene PsaA may be helpful to the invasion of SP to the blood system.

[MALDI-TOF-MS combined with WCX technique in diagnosing pituitary adenoma].

OBJECTIVE: Application of matrix assisted laser desorption ionization time of flight mass spectrometry enhancement (MALDI-TOF-MS) combined with WCX nanometer magnetic bead technique, screening of the serum biomarkers in pituitary adenoma, to establish a serum protein fingerprint classification decision tree. METHODS: Analyse the serum samples of 40 cases of pituitary adenoma and 60 cases of healthy adult and find the different protein peaks, then to establish the diagnosis model and the classification decision tree of pituitary adenomas. RESULTS: A total of 42 differences in protein peaks were identified in the experimental and control group (P < 0.01). The diagnosis model of pituitary adenomas was established by three protein peaks (3382.0, 4601.9, 9191.2). The model could screen the pituitary adenoma out of the normal population. The sensitivity was 90.00% and the specificity was 88.30%. By the double blind experimental validation, the model could diagnose the pituitary adenoma and the sensitivity was 88%, the specificity was 83.30%. CONCLUSION: Significantly different protein peaks can be screened out between pituitary adenoma cases and healthy controls using MALDI-TOF-MS combined with WCX technique, and these protein peaks may be used as a pituitary adenoma detection, follow-up indicator.

Pituitary adenoma biomarkers identified using proteomic fingerprint technology.

OBJECTIVE: To determine whether pituitary adenomas can be diagnosed by identifying protein biomarkers in the serum. METHODS: We compared serum proteins from 65 pituitary adenoma patients and 90 healthy donors using proteomic fingerprint technology combining magnetic beads with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). RESULTS: A total of 42 M/Z peaks were identified as related to pituitary adenoma (P<0.01). A diagnostic model established based on three biomarkers (3382.0, 4601.9, 9191.2) showed that the sensitivity of diagnosing pituitary adenoma was 90.0% and the specificity was 88.3%. The model was further tested by blind analysis showing that the sensitivity was 88.0% and the specificity was 83.3%. CONCLUSIONS: These results suggest that proteomic fingerprint technology can be used to identify pituitary adenoma biomarkers and the model based on three biomarkers (3382.0, 4601.9, 9191.2) provides a powerful and reliable method for diagnosing pituitary adenoma.

SELDI-TOF MS combined with magnetic beads for detecting serum protein biomarkers and establishment of a boosting decision tree model for diagnosis of pancreatic cancer.

AIM: New technologies for the early detection of pancreatic cancer (PC) are urgently needed. The aim of the present study was to screen for the potential protein biomarkers in serum using proteomic fingerprint technology. METHODS: Magnetic beads combined with surface-enhanced laser desorption/ionization (SELDI) TOF MS were used to profile and compare the protein spectra of serum samples from 85 patients with pancreatic cancer, 50 patients with acute-on-chronic pancreatitis and 98 healthy blood donors. Proteomic patterns associated with pancreatic cancer were identified with Biomarker Patterns Software. RESULTS: A total of 37 differential m/z peaks were identified that were related to PC (P<0.01). A tree model of biomarkers was constructed with the software based on the three biomarkers (7762 Da, 8560 Da, 11654 Da), this showing excellent separation between pancreatic cancer and non-cancer., with a sensitivity of 93.3% and a specificity of 95.6%. Blind test data showed a sensitivity of 88% and a specificity of 91.4%. CONCLUSIONS: The results suggested that serum biomarkers for pancreatic cancer can be detected using SELDI-TOF-MS combined with magnetic beads. Application of combined biomarkers may provide a powerful and reliable diagnostic method for pancreatic cancer with a high sensitivity and specificity.

The expression of cyclin G in nasopharyngeal carcinoma and its significance.

To investigate the expression of cyclin G1, cyclin G2 in nasopharyngeal carcinoma (NPC) cell lines and its significance. The protein expression of cyclin G1, cyclin G2 in NPC cell lines of different differentiation degree (HNE2, CNE1) was detected by indirect immunofluorescence. The mRNA expression of cyclin G1, cyclin G2 in HNE2 and CNE1 was measured with RT-PCR. The cyclin G1 expression in HNE2 and CNE1 was weak, and cyclin G2 expression in the cytoplasm near cell membrane was strong, continuous, and homogeneous. The expression level of cyclin G1-mRNA in HNE2 was 2.097 ± 0.262, which was significantly higher than CNE1 (0.997 ± 0.286, P < 0.05); the expression level of cyclin G2-mRNA in HNE2 was 0.708 ± 0.107, which was significantly lower than CNE1 (1.216 ± 0.037, P < 0.05). Abnormal expression of cyclin G was closely related to tumor differentiation, the origin, and progression of NPC.

New serum biomarkers for detection of tuberculosis using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry.

BACKGROUND: New technologies for the early detection of tuberculosis (TB) are urgently needed. Pathological changes within an organ might be reflected in proteomic patterns in serum. The aim of the present study was to screen for the potential protein biomarkers in serum for the diagnosis of TB using proteomic fingerprint technology. METHODS: Proteomic fingerprint technology combining protein chips with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) was used to profile the serum proteins from 50 patients with TB, 25 patients with lung disease other than TB, and 25 healthy volunteers. The protein fingerprint expression of all the serum samples and the resulting profiles between TB and control groups were analyzed with the Biomarker Wizard system. RESULTS: A total of 30 discriminating m/z peaks were detected that were related to TB (p<0.01). The model of biomarkers constructed by the Biomarker Patterns Software based on the three biomarkers (2024, 8007, and 8598 Da) generated excellent separation between the TB and control groups. The sensitivity was 84.0% and the specificity was 86.0%. Blind test data indicated a sensitivity of 80.0% and a specificity of 84.2%. CONCLUSIONS: The data suggested a potential application of SELDI-TOF MS as an effective technology to profile serum proteome, and with pattern analysis, a diagnostic model comprising three potential biomarkers was indicated to differentiate people with TB and healthy controls rapidly and precisely.

[Detection of differentially expressed proteins in the semen of oligospermia patients by SELDI-TOF-MS technology].

OBJECTIVE: To analyze the spectrometric semen protein profiling of oligospermia patients and healthy controls by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS), and to establish a semen marker pattern for the diagnosis of oligospermia. METHODS: Semen samples of 33 oligospermia patients and 31 healthy controls were collected on the CM01O proteinchip. The spectrometric protein profiling was detected by SELDI-TOF-MS and the data analyzed by Biomarker Pattern Software provided by Ciphergen Corp. A primary diagnostic model of oligospermia was established and evaluated by blind test with the 33 patients and 31 healthy controls. RESULTS: A total of 185 protein peaks were detected at the molecular range of 2000-20,000, among which 23 showed significant differences between oligospermia patients and healthy controls (P < 0.05). The diagnostic model consisted of 3 protein peaks, with a sensitivity of 90.9% (30/33) and a specificity of 93.6% (29/31). And the double-blind test generated a sensitivity of 87.8% (29/33) and a specificity of 90.3% (28/31). CONCLUSION: The diagnostic model was successfully established by SELDI-TOF-MS, which could be applied to the differentiation of the spectrometric protein profiling patterns of oligospermia patients and healthy controls.

Serum proteomic patterns for gastric lesions as revealed by SELDI mass spectrometry.

SELDI mass spectrometry was used to investigate protein expression in sera of patients with gastric cancer and gastritis compared to normal volunteers. Differences in peak morphology and intensity were observed in regions of 5910 Da, 5084 Da, 6640 Da and 8691 Da. Patients with gastric cancer exhibited an up-regulation of the 5910 Da peak and a down-regulation of the 8691 Da peak compared to the healthy volunteers; there was also some bi-partitioning and tri-partitioning at the 5084 Da peak. When comparing the sera of these cancer patients with those of gastritis, the former had an up-regulation of the 5910 Da peak and a down-regulation of the 6640 Da peak. This is the first report showing that SELDI sera analysis may be useful in the screening of gastric lesions.