[LC-MS/MS Analysis on the Components of Excretory-secretory Protein of Trichinella spiralis Muscle Larvae].

Objective: To analyze the components of excretory-secretory protein（ESP） of Trichinella spiralis muscle larvae, and search for the anti-tumor protein components. Methods: The Trichinella spiralis muscle larvae were collected, and ESP was prepared. The ESP was separated in 15% SDS-PAGE. Proteins extracted from the protein bands were lysed with trypsin, and analyzed by LC-MS/MS. The identified proteins were classified by Gene Ontology（GO） according to cell component, molecular function, and biological processes. Results: SDS-PAGE revealed clear protein bands at Mr 10 000-142 000. A total of 162 proteins were analyzed with LC-MS/MS, of which 63 were identified, 34 were putative proteins, and 65 were unidentified proteins. Six anti-tumor relevant proteins were revealed, which were tropomyosin, histone H2A, cleavage and polyadenylation specificity factor subunit 2, serine proteinase inhibitor Kazal-type 4, Armadillo segment polarity protein and eukaryotic initiation factor 4A. The GO enrichment analysis showed that the identified proteins possessed 54 different types of molecular functions, and participated in cell structure and 382 biological processes. Conclusion: The ESP of Trichinella spiralis muscle larvae has complex protein components, many with unknown identities. Six anti-tumor relevant proteins were determined from the 63 identified proteins.

[Effect of the excretory/scretory proteins from Trichinella spiralis on apoptosis of NCI-H446 small-cell lung cancer cells].

OBJECTIVE: To investigate the effect of excretory/secretory proteins from Trichinella spiralis on apoptosis of NCI-H446 small-cell lung cancer cells. METHODS: Trichinella spiralis muscle stage larvae (5 x 10(6)/ml) were cultured in culture media for 24 h, the excretory/secretory proteins were collected from the supernatant of culture media. NCI-H446 small-cell lung cancer cells (No. A05) were randomly divided into three groups: experiment group (A), standard control group (apoptosis group, B), and control group (C). NCI-H446 cells in groups A and B were cultured with 0.3 mg/ml T. spiralis excretory/secretory proteins, and 6.4 microg/ml cisplatin for 24 h, respectively. NCI-H446 cells of group C were cultured for 24 h without any treatment. The expression of Bcl-2, Fas and Fasl mRNA was detected by RT-PCR. C-myc protein expression level was examined by Western blotting and immunofluorescence assay. RESULTS: The level of Bcl-2 mRNA was lowest in group A(0.575 +/- 0.047) , Bcl-2 mRNA level in group C (0.975 +/- 0.069) was higher than that of group B (0.850 +/- 0.073) (P<0.05). Fas mRNA level was highest in group A (0.975 +/- 0.115), followed by group B (0.817 +/- 0.121) and group C(0.769 +/- 0.061) (P<0.05). The level of Fasl mRNA in groups A, B, and C was 0.669 +/- 0.051, 0.787 +/- 0.124, and 0.875 +/- 0.125, respectively (P<0.05). Fas/Fasl mRNA ratio in groups A, B, and C was 1.475, 1.038, and 0.878. Western blotting showed that the expression of C-myc protein in group C (1.172 +/- 0.026) was highest, followed by group B (1.074 +/- 0.069) and A (0.566 +/- 0.054) (P<0.05). Immunofluorescence test indicated that the C-mye protein was found in the cytoplasm and the nucleus 24 h after treated with 0.3 mg/ml T. spiralis excretory/secretory proteins and 6.4 p.g/ml cisplatin. CONCLUSION: Trichinella spiralis excretory/secretory proteins may inhibit apoptosis of NCI-H446 small-cell lung cancer cells by reducing the apoptosis protein C-myc and Bcl-2 mRNA levels, and causing the increase of Fas/Fasl mRNA ratio.

Free-radical oxidation and superoxide dismutase activity in synovial fluid of patients with temporomandibular disorders.

AIMS: To measure the activity of oxygen free radicals and the level of antioxidant enzyme superoxide dismutase (SOD) in the synovial fluid (SF) of the temporomandibular joints (TMJs) of patients with temporomandibular disorders (TMD). METHODS: Thirty-two patients were divided into 3 subgroups: anterior disc displacement with reduction, anterior disc displacement without reduction, and osteoarthrosis. Six healthy volunteers served as controls. A pumping procedure was used to take SF from the superior TMJ space. The concentration of lipid peroxidation products was assessed by means of the thiobarbituric acid reaction, and the level of SOD was assessed with spectrophotometry. RESULTS: SF levels of lipid peroxides (LPO) in the control and patient groups were 0.010 (+/- 0.004) x 10(-3) nmol/mg protein and 0.61 (+/- 0.121) x 10(-3) nmol/mg protein, respectively. SF SOD levels were 4.61 (+/- 1.30) NU/mg protein and 9.83 (+/- 2.66) NU/mg protein, respectively. Both the concentration of LPO and the level of SOD activity were significantly higher in the TMD patients than in the normal control subjects (P < .001 and P < .01, respectively). There were no significant differences in the level of LPO or SOD between the 3 subgroups. CONCLUSION: These results demonstrate oxygen free radicals and antioxidant enzymes may be connected in the pathogenesis of TMD.