Objective To find early diagnostic biomarkers for colorectal carcinoma by comparing differential(expressing) proteins from colorectal carcinoma and normal colorectal tissues.Methods Colorectal carcinoma tissues and paired normal tumor-adjacent colorectal tissues were collected,and tissue total protein was (extracted);differential proteome profiles were established and analysed by means of immobilized pH(gradient-based) two-dimesional polyacrylamide gel electrophoresis(2D-PAGE) and matrix-assisted laser(desorption)/ionization time of flight mass spectrometry(MALDI-TOF-MS).Results Well-resolved,(reproducible) 2-DE profiles of human colorectal carcinoma tissues and paired normal tumor adjacent colorectal tissues were obtained.For tumor tissue,a total of 1098?28 spots were detected,and for normal tissue,760?45 spots were detected.For normal tissue,The average deviation of spot position was(0.542?(0.12))mm in IEF direction and(0.933?0.098)mm in SDS-PGE direction for tumor tissue.The average deviation of spot position was(0.745?0.130)mm in IEF direction and(1.233?0.272)mm in(SDS-PGE) direction.30 differential expressing proteins were analysed by mass spectrometry and bioinformation,16 of them were well characterized including Apolipoprotein A1(apoA1),calreticulin precursor,glutathione(S-transferase),hepatic fatty acid-binding protein、heat shock protein 27 ect.Conclusions Differential expression proteins can be candidate biomarkers for early diagnosis of colorectal carcinoma;and proteomic technique is valuable for screening the diagnostic biomarkers.

OBJECTIVETo study the dynamic changes in the protein marker expression in the spermatogonial stem cells (SSCs) of mice at different ages by iTRAQ protein mass spectrometry and to screen new markers using the bioinformatic proteome database.

METHODSBased on the postnatal weeks, we divided 80 healthy male C57BL/6 mice into eight age groups of equal number, harvested their testicular tissues, extracted proteins following purification of the SSCs by compound enzyme digestion and magnetic-activated cell sorting. Then we analyzed and identified proteins using two-dimensional electrophoresis, protein mass spectrometry, and protein database information.

RESULTSTotally, 248,510 mass spectra were obtained from the MS experiment and 1132 proteins were identified. By the criteria of >1.2-fold for protein abundance difference and P value <0.05, we identified 298 differentially expressed proteins and 9 currently known makers of SSCs (PCNA, GFRalpha1, CDH1, Annexin A7, UCHL1, VASA, CD49f, CD29, and PLZf). Compara- tive analysis showed different expressions of the proteins in the SSCs of the mice of different ages, and the differences in the expressions of GFRalpha1, CD49f, and CD29 were consistent with the findings in other published literature. Ten proteins (P63, CD71, CD98, K19, ACE, K18, K15, K17, SH2, and SH3) were selected as SSC markers to be further studied.

CONCLUSIONThe proteins in SSCs are differentially expressed in mice of different ages. The technology of iTRAQ protein mass spectrometry can be used to analyze and compare the proteome information of mouse SSCs, obtain differentially expressed proteins in mice of different ages, and thus offers a new ap- proach to further analysis and study of the function and roles of these differential proteins.

Adult Stem Cells ; cytology ; metabolism ; Age Factors ; Animals ; Biomarkers ; analysis ; metabolism ; Cell Separation ; methods ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Proteins ; analysis ; metabolism ; Spermatogonia ; cytology

Objective To explore the effects of ethyl-3,4 dihydroxybenzoate(EDHB), a prolyl hydroxylase inhibitor, pretreatment on the tubular epithelial cells apoptosis induced by albumin and hypoxia. Methods To investigate the effects of albumin and hypoxia on cells, rat tubular epithelial cells(NRK-52E)were incubated for 24 h in:(1)normoxia(5%CO2);(2)hypoxia(1% O2);(3)albumin(30 g/L)under normoxia;(4)albumin(30 g/L)under hypoxia. To investigate the effects of EDHB pretreatment on cells apoptosis, NRK-52E were incubated in hypoxia for 24 h in:(1)normoxia;(2)hypoxia;(3)hypoxia+albumin(30 g/L);(4)hypoxia+EDHB(500 μmol/L);(5)EDHB pretreatment(albumin 30 min after EDHB). Apoptosis was measured by flow cytometry(AnnexinV-FITC-PI). bcl-2, bax and vascular epithelial growth factor(VEGF)mRNA expression were detected by RT-PCR. VEGF protein expression was detected by Western blotting. Results NRK-52E apoptosis was not significantly different between hypoxia and norraoxia groups(P>0.05), but increased significantly in albumin(30 g/L)under hypoxia group compared with albumin(30 g/L)under normoxia group(37.36%?.95% vs 25.59%?.32%, P< 0.05). There was an increase in bax mRNA expression and a decrease in bcl-2 mRNA expression in albumin(30 g/L)under hypoxia group compared with albumin(30 g/L)under normoxia group(P< 0.05). EDHB pretreatment improved these impairments of albumin(30 g/L)under hypoxia on NRK-52E(P< 0.05). VEGF expression elevated in hypoxia compared with normoxia(P<0.05), decreased in albumin(30 g/L)under hypoxia groups compared with that without albumin groups(P<0.05).EDHB pretreatment significantly improved VEGF expression compared with albumin(30 g/L)under hypoxia group(P <0.05). Conclusion NRK-52E cells apoptosis induced by albumin is accelerated by hypoxia, however partially improved by EDHB pretreatment, probably through the up-regulation of VEGF expression.

Objective To provide molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer. Differentially expressed proteins in left sided colon cancer and right sided colon cancer were screened by proteomic technique. Methods Tissue samples including left sided colon cancer and right sided colon cancer were collected and preserved in the -80℃ refrigerator. In the first part of our experiment, protein was separated by 2-dimensional gel electrophoresis (2-DE) and the images of the gels were acquired by the scanner and then analyzed to find the differentially expression protein-spots in different groups. The peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorptiorn/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching in the Mascotdatabase. Differentially expressed proteins were assayed by RT-PCR, Western blot, and immunohistochemical method. Results Altogether 55 differentially expressed protein spots were screened and 21 spots of them were identified. Compared with the right sided colon cancer, 14 proteins were up-regulated and 7 proteins down-regulated including HSP27 in the left sided colon cancer. HSP27 expressed higher in the right sided colon cancer than in the left sided colon cancer.Conclusion There are differentially expressed proteins in left sided colon cancer and right sided colon cancer, especially difference in HSP27 expression at mRNA and protein level, which may be molecular genetic basis for oncobiological difference in left sided colon cancer and right sided colon cancer.

Knee osteoarthritis is one of the common type of arthropathy, the clinical stage of the typical patients belongs to the middle-late stage, so it urges to improve the early diagnosis. At present, magnetic resonance imaging is most used in clinical diagnosis of knee osteoarthritis, and with the development of different MRI sequences, the sequences of early articular cartilage lesions are used in clinic. In the early diagnosis of knee osteoarthritis, the simple and practical methods such as ultrasonography is becoming a trend, and the specific biomarkers of early knee osteoarthritis have become the hot research. This overview article outlined the methods of early diagnosis from the ultrashort echo time MRI, ultrasonography and biomarkers.
Biomarkers ; analysis ; Early Diagnosis ; Humans ; Magnetic Resonance Imaging ; Osteoarthritis, Knee ; diagnosis ; diagnostic imaging ; metabolism ; Radiography ; Ultrasonography

OBJECTIVE To study the toxicological effect of matrine (MT) on PC12 cells and the mechanism of mitochondrial damage. METHODS After treatment with MT 2, 4 and 8 mmol·L-1for 8, 16,24 and 48 h,respectively,the viability of PC12 cells was measured with methylthiazolyltetrazolium assay.PC12 cells were treated with MT 2,4 and 8 mmol·L-1for 24 h,before the morphological changes were observed by Hoechst33342 staining. The superoxide dismutase (SOD) activity and malondialde-hyde (MDA) content in cells were detected using hydroxylamine method and thiobarbituric acid method, apoptosis rate and reactive oxygen species (ROS) of PC12 cells were detected with flow cytometry, mitochondrial membrane potential (MMP) was detected with JC-1 staining, and the expressions of procaspase 3, procaspase 9, cleaved-caspase 3, Bax and Bcl-2 were detected with Western blotting. RESULTS MT inhibited the growth of PC12 cells in a time-and concentration-dependent manner.After being treated with MT for 24 h,the nuclei of PC12 cells in MT groups showed chromatin agglutination and partial rupture to different degrees,and compared with normal control group,the apoptosis rates of MT 2,4 and 8 mmol·L-1groups were significantly increased(P<0.01).Intracellular ROS and MDA levels increased (P<0.05, P<0.01), SOD activity decreased (P<0.01), and MMP decreased. The expressions of procaspase 9, procaspase 3 and Bcl-2 were significantly decreased (P<0.01, P<0.05), the expres-sions of cleaved-caspase 3 and Bax were significantly increased(P<0.05,P<0.01),and the ratio of Bcl-2/Bax was significantly decreased (P<0.01). CONCLUSION MT has toxic effect on PC12 cells and induces apoptosis through ROS mediated mitochondrial pathway.

Objective To study the effects of granisetron hydrochloride on vasovagal syncope(VVS) in rabbits.Methods Twenty-four healthy New Zealand rabbits were divided stochastically into control group and intervention group,12 in each group. The control group was injected intravenously with normal saline. The intervention group was injected intravenously with granisetron hydrchloiride.Rabbit VVS models were established,each was taken at 4 points in time in the bloodletting process:T1,T2,T3,T4,to compare the bloodletting time,the concentration of 5-hydroxytryptamine(5-HT) in T2,T3,T4 and the total blood volume between the groups,and monitor the heart rate, blood pressure during the entire process.Results 1.The time of intervention group in T2,T3,T4 was longer than the time of control group obviously(P

OBJECTIVETo analyze the differential protein expression of left-sided colon cancer and right-sided colon cancer.

METHODSTissue samples of left-sided colon cancer (n=7) and right-sided colon cancer (n=7) were collected. Tissue protein was abstracted and two-dimensional gel electrophoresis (2-DE) was used to examine the gel images. Peptide mass fingerprintings (PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and the proteins were identified by data searching with bioinformatics. Immunohistochemical SP method was used for the detection of glucose regulated protein 78 kD (GRP78) and heat shock protein 20 (HSP20) in left-sided colon cancer (n=50) and right-sided colon cancer (n=50) tissues.

RESULTSSixteen differentiating protein spots were identified. Compared with right-sided colon cancer, 10 proteins including GRP78 up-regulated and 6 proteins including HSP20 down-regulated in left-sided colon cancer. Immunohistochemical detection showed that in left and right sided colon cancer, the positive expression rate of GRP78 was 78% (39/50) and 56% (28/50) and the positive expression rate of HSP20 was 34% (17/50) and 72% (36/50), respectively, and the differences were statistically significant (both P<0.05). The positive rate of GRP78 was associated with tumor differentiation, infiltration layer, TNM staging, lymph node metastasis, and liver metastasis, while the positive rate of HSP20 was associated with tumor gross morphology, TNM staging, and lymph node metastasis (P<0.05).

CONCLUSIONSThere are differentially expressed proteins between left-sided colon cancer and right-sided colon cancer, especially for GRP78 and HSP20, which may be the cause leading to the biological differences between left-sided and right-sided colon cancer.

Adult ; Aged ; Colonic Neoplasms ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; HSP20 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Middle Aged

Intervertebral disc degeneration is considered as a primary cause of clinical low back pain, however the molecular mechanism is not clear yet. Recently, researches on the molecular basis of intervertebral disc degeneration have become a hotspot. The special structure and biomechanics properties of the disc contribute to its propensity toward degeneration. Intervertebral disc degeneration is associated with the changes of the cytological behavior,including the increase in cell death and the degradation of extracellular matrix. However, the mechanism of cell death including cell apoptosis and autophagy in intervertebral disc degeneration remains unclear. Further study on the molecular mechanism of intervertebral disc degeneration is the foundation of improving and treating the intervertebral disc degeneration in the future. Although some progresses are made in the aspect of biological study, the biological environment of intervertebral disc itself is still a challenge for the development of biological treatment. This article is to review the latest advance on the biological characteristics of normal intervertebral disc and the cell death in the process of the intervertebral disc degeneration.
Animals ; Apoptosis ; Cell Death ; Extracellular Matrix ; metabolism ; Humans ; Intervertebral Disc ; cytology ; metabolism ; Intervertebral Disc Degeneration ; metabolism ; physiopathology