Background The circulating endothelial progenitor cells (EPCs)play an important role in postnatal vasculogenesis and restoration of endothelial injury.Previous investigation illustrated that a reduced ocular blood flow and vascular dysfunction caused by endothelial dysfunction plays a role in the pathogenesis of glaucoma.However,endothelial system accommodation is accomplished with the circulating EPCs.Objective The present trail was to investigate EPCs change in patients with primary acute angle-closure glaucoma (PACG)and explore the role of EPCs in the pathogenesis of PACG. MethodsA prospective cohort study was designed.Thirty patients with PACG were enrolled in Tianjin Medical University General Hospital as PACG group,and 20 normal subjects served as control group.Periphery bloodsamples were obtained fromall the patients and thenstained with saturating concentrations of monoclonal antibodies,FITC-conjugated anti-CD34 and CD133 mAb.EPCs identified by CD34,CD133 were enumerated by flow cytometry,and the correlation between EPCs change and its relative factors was analyzed.Informed consent was obtained from each subject before any medical procedure. Results No significant differences were found in age,gender,vascular-related risk factor,blood biochemical indicators between PACG group and normal contol group(P＞0.05 ),but a higher intraocular pressure( IOP)was displayed in PACG group compared with normal control group ( P =0.00 ).The numbers of EPCs were ( 48 ± 22 ) cells/ml in PACG group and ( 65 ± 20 )cells/ml in normal control group with a significant difference between them (P=0.004).In PACG group,the numbers of EPCs were(60± 19 )cells/ml and (34 ±7 )cells/ml respeetively in phase 1 and phase 3 of optical nerve damage (Z=-3.015,P=0.002 ).There was a negative correlation between EPCs numbers and baseline IOP within a certain range( r=-0.835,P＜0.05 ).However,no obvious correlations were seen between EPCs numbers and blood lipid Level,blood glucose level or glaucoma course ( r =0.343,P =0.227 ; r =-0.203,P =0.419 ; r =0.198,P =0.610 ).The EPCs numbers in PACG patients with cardiovascular disease were(56±22)cells/ml and that of without PACG were (35± 15 ) cells/ml( P =0.005 ). ConclusionsThe numbers of EPCs decrease in PACG patient.These results imply that EPCs might play role during the restore of the optical nerve damage in PACG eye.

BackgroundLaser peripheral iridotomy(LPI) can break the pupillary block,and is an effective method of treating acute primary angle closure (APAC).However,a part of APAC eyes may gradually develop a formation and extension of peripheral anterior synechia(PAS) and increased intraocular pressure(IOP) after LPI.ObjectiveTo investigate the relationship between appositional angle closure and darkroom provocative test(DRPT) in the fellow eyes with APAC after LPI.Methods Fellow eyes of APAC without PAS after LPI were studied.Ultrasounic biomicroscopy(UBM) were performed in darkness to observe whether appositional angle closure occurred and compare the relationship between the quadrants with appositional angle closure and the results of DRPT.Results Fifty-four patients were included in the study.Appositional angle closure was observed in at least one quadrant in 20(37.0％ ) of the 54 fellow eyes with APAC after LPI.Fifty-one patients were given DRPT and positive result in 9 patients( 17.6％ ).According to the quadrants with appositional angle closure,there were 5 patients with DRPT positive results in 46 patients with appositional angle closure 0 to 2 quadrants,and 4 patients with DRPT positive results in 5 patients with appositional angle closure 3 to 4 quadrants ( P =0.003 ).Bivariate correlation analysis indicated a positive correlation between the value of the increased IOP in DRPT and the number of quadrants with appositional angle closure in darkness( r =0.397,P =0.004).ConclusionsA certain proportional fellow eyes of APAC appeared appositional angle closure in darkness and DRPT positive result after LPI.The more the quadrants of appositional angle closure after LPI,the greater the likelihood of a positive DRPT.It suggests that the APAC fellow eyes and attack eyes with the same anatomical configuration still have the possibility of angle closure after LPI,and need follow-up and treatment for a long time.

Primary angle closure glaucoma(PACG)is the leading type of glaucoma in Asia, especially in China. PACG still has a high proportion of angle closure after laser peripheral iridectomy(LPI). Plateau iris is one of the non-pupillary blockage factors that cause angle closure. With the wide application of ultrasound biomicroscopy(UBM)in ophthalmology, the understanding of plateau iris has been deepened continually. This paper will elaborate the concept, mechanism, prevalence, relationship with angle closure, diagnostic criteria and the treatment of plateau iris, aiming to have a deeper understanding of the relationship between plateau iris and PACG, and to provide references for the treatment and research of PACG in the future.

OBJECTIVETo study the expression pattern of peroxisome proliferator-activated receptor (PPAR) pathway molecules in rat lung tissue under hyperbaric oxygen exposure.

METHODSTwenty seven male SD rats were randomly divided into hyperbaric normoxia group (0.23 MPa air), hyperbaric oxygen treatment time series group (0.23 MPa oxygen, were exposed for 2 h, 4 h, 6 h or 8 h), continuous small flow of ventilation to maintain cabin O2 concentration > 99%. HE staining of lung tissue morphological changes and application oligo microarray to each time point lung were observed. Part of the PPAR pathway genes were validated by RT-PCR.

RESULTSCompared with hyperbaric normoxia group, the lung injury caused by hyperbaric oxygen treatment gradually deteriorated during the time series. Expression microarray analysis of gene ontology (Go) enrichment analysis results in a class of PPAR pathway class included multiple PPAR pathway molecule. RT-PCR results suggested that PPAR-8 and PPAR-Y were up-regulated in the lung tissue after a long time exposure to hyperbaric oxygen.

CONCLUSIONPro-longed hyperbaric oxygen exposure causing pulmonary oxygen toxicity can induce the activation of the PPAR pathway.

Animals ; Hyperbaric Oxygenation ; adverse effects ; Lung ; metabolism ; pathology ; Male ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

To study the effects of tyrosine-kinase inhibitor STI571 on the adhesion of RPMI8226 cells to fibronectin (FN), cell adhesion mediated adriamycin-resistance and the Rac1 mRNA expression, the adhesion of RPMI8226 cells to fibronectin and drug resistance mediated by cell adhesion were determined by means of crystal violet staining and MTT assays respectively, Rac1 mRNA levels in RPMI8226 cells were examined by semi-quantitative RT-PCR. The results showed that STI571 could inhibit the adhesion of RPMI8226 cells to fibronectin. When RPMI8226 cells had been adhered to FN or BSA-coated wells for 1, 6 and 12 hours, the adhesion rates were (43.71 +/- 2.18)%, (55.63 +/- 1.56)%, and (63.42 +/- 2.46)% respectively. After treatment with STI571 20 micromol/L, the adhesion rates decreased to (15.12 +/- 1.04)%, (17.58 +/- 1.32)% and (17.24 +/- 1.59)% respectively (P < 0.05). The experiment revealed that growth of RPMI8226 cells adhered to FN-coated plates had a significant advantage over growth on BSA-coated plates when exposed to adriamycin (Adr) for 1 hour followed by a 24-hour culture period, and the mean IC(50) value for FN-adhered cells was (1.46 +/- 0.04) micromol/L while mean IC(50) value for BSA control was (0.78 +/- 0.03) micromol/L (P < 0.05). Following treatment with 20 micromol/L STI571, the mean IC50 values for FN and BSA adhered cells were (0.81 +/- 0.05) micromol/L, (0.74 +/- 0.02) micromol/L respectively, there was no significant difference between them (P > 0.05). RT-PCR demonstrated that the relative Rac1 mRNA level (Rac1/GAPDH) in RPMI8226 cells was downregulated following being treated with 20 micromol/L STI571. It is concluded that STI571 can inhibit the adhesion of RPMI8226 cells to fibronectin, reverse cell adhesion mediated adriamycin-resistance, and downregulate Rac1 mRNA level.
Benzamides ; Cell Adhesion ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Fibronectins ; metabolism ; Humans ; Imatinib Mesylate ; Multiple Myeloma ; metabolism ; pathology ; Piperazines ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured ; rac1 GTP-Binding Protein ; biosynthesis ; genetics

Cytomegalovirus (CMV) infection is a dangerous complication in patients with chronic graft versus host disease (cGVHD). CMV-specific immunity depends on the activity of T cells. This study was aimed to investigate the effect of CMV pp65 gene modified dendritic cells (DCs) on activation of autologous T cells. Lentivirus system was utilized to introduce the CMV full-length pp65 gene into mouse DCs; CpG-DNA was used to induce mature DCs; flow cytometry and immunofluorescence were used to determine the expression of antigen and IFNgamma in T lymphocytes. The results showed that the DCs were infected with lentivirus at a multiplicity of infection (MOI) of 50 with optimal infectious efficiency of 30%-40%; mature DCs expressing pp65 gene could stimulate autologous naive T cells to express CD69 specifically; mature DCs expressing PP65 could stimulate autologous CD4+ or CD8+ T cells to produce IFNgamma. It is concluded that CMV pp65-modified and CpG-DNA-induced mature DCs can activate CMV-specific T lymphocytes in vitro.
Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; genetics ; metabolism ; Antigens, Viral ; immunology ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; CpG Islands ; genetics ; Cytomegalovirus ; immunology ; DNA ; genetics ; Dendritic Cells ; cytology ; immunology ; metabolism ; Humans ; Interferon-gamma ; genetics ; metabolism ; Lectins, C-Type ; Lentivirus ; genetics ; metabolism ; Mice ; Phosphoproteins ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism

OBJECTIVETo investigate the expressions of matrix metalloprotein-2 (MMP-2) and tissue inhibitor of metallopeptidase inhibitor-1 (TIMP-1) in the renal allografts of patients with chronic active antibody-mediated rejection (ABMR), and explore their role in the pathogenesis of ABMR.

METHODSImmunohistochemistry and computer-assisted image analysis were used to detect the expression of MMP-2 and TIMP-1 in the renal allografts of 46 patients with interstitial fibrosis and tubular atrophy (IF/TA), with 15 normal renal tissue specimens as the control. The association of MMP-2 and TIMP-1 with the pathological grade of IF/TA in ABMR was analyzed.

RESULTSThe expressions of MMP-2 and TIMP-1 significantly increased in the renal tissues of the patients as compared with the normal renal tissues (P<0.05). MMP-2 expression tended to decrease, while TIMP-1 and serum creatinine increased with the pathological grades of IF/TA (P<0.05). In IF/TA group, the expression of TIMP-1 was positively correlated to serum creatinine level (r=0.718, P=0.00<0.05).

CONCLUSIONAbnormal expressions of MMP-2 and TIMP-1 can promote the development of renal fibrosis in chronic ABMR.

Adult ; Antibody Formation ; Complement C4b ; metabolism ; Female ; Fibrosis ; etiology ; Graft Rejection ; immunology ; Humans ; Kidney ; metabolism ; Kidney Diseases ; pathology ; Kidney Transplantation ; adverse effects ; immunology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Peptide Fragments ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

Differential proteomics analysis of Sika deer antlers at rapid growth stage (60 d) and ossification stage ( 90 d) was performed by isobaric tags for relative and absolute quantitation ( iTRAQ ) , ultra high performance liquid chromatography and mass spectrometry technologies. A total of 127 differential proteins were identified. Compared with the ossification stage, 80 differential proteins were significantly up-regulated and 47 differential proteins were significantly down-regulated at the rapid growth stage. These differential proteins were mainly distributed in the regions of extracellular matrix, nucleosome, haptoglobin-hemoglobin complex, actin filament, endoplasmic reticulum-Golgi intermediate compartment, endoplasmic reticulum lumen, and endometrium, etc. The up-regulated differential proteins were mainly involved in the regulations of oxygen transport in the blood, nerve growth and regeneration, cartilage and bone development and ATP synthesis compared with ossification stage, and the down-regulated differential proteins were mainly involved in the endochondral ossification process. The changes of protein expression at different growth stages were closely related to antler rapid growth and ossification. Therefore, the results of this study provided a basic data for discovering the molecular mechanisms of antler rapid growth and ossification, and it was of great significance for further study of the pharmacological basis and clinical application of antlers.

Objective To explore the imaging and clinical characteristics and related risk factors of patients with coronary artery stenosis located proximally to myocardial bridging.Methods This study enrolled 603 patients with angiography evidenced myocardial bridging-mural coronary artery between May 2004 to May 2009.Angiographic and clinic data were collected according to uniform protocol and standard questionnaires were used to obtain patients' demographic and clinical information.Univariate and multivariate analysis were performed to explore related risk factors.Results Chest pain was present in 247cases (41.0％).Dynamic ST-T changes were found in 229 cases (38％).A total of 644 myocardial bridging-mural coronary arteries were detected including 382 (62.4％) segments located proximally to myocardial bridging.Diastolic vessel diameters in the myocardial bridging segment were significantly smaller than reference segments (all P ＜ 0.01).Stepwise multiple regression analysis suggested that vascular bifurcation lesions,the degree of narrowing and the number of diseased coronary vessels of non-myocardial bridging-mural coronary arteries,age,LDL-C/HDL-C,male gender,diabetes,and systolic narrow rate of myocardial bridging-mural coronary arteries were positively related with the narrowing degree of the first coronary artery stenosis located proximally to myocardial bridging (P ＜ 0.05 or P ＜ 0.01).Vascular bifurcation lesions,the degree of narrowing and the number of diseased coronary vessels of non-myocardial bridging-mural coronary arteries,age,LDL-C/HDL-C,male,diabetes and dyslipidemia were positively related with the narrowing degree of the most severe coronary artery stenosis located proximally to myocardial bridging(P ＜ 0.05 or P ＜ 0.01).Conclusions Myocardial ischemia is common in patients with myocardial bridging and the artery segments located proximally to myocardial bridging are prone to stenosis.Systolic narrow rate of myocardial bridging-mural coronary arterics is one of major determinants of coronary artery stenosis located proximally to myocardial bridging.Whereas the other coronary heart disease risk factors are likely to play more important roles.

Objective To investigate effects of different de-cellularization methods on biomechanical properties and histological structure of annulus fibrosus in pigtails and provide experimental evidence for the construction of tissue engineering annulus fibrosus. Methods Sixty Fresh annulus fibrosus were dissected from caudal disks of pigs and randomly assigned to 4 groups with 15 in each group. Triton X-100 group(Group A): annulus fibrosus were treated with hypotonic Tris-HCl buffer for 48 hours and de-cellularized with Triton X-100, DNase Ⅰ and RNase A. SDS group (Group B): annulus fibrosus were subjected to 3 cycles of freeze-thaw and subsequently de-cellularized with SDS, DNaseⅠ and RNase A. Trypsin group (Group C): annulus fibrosus were de-cellularized with Tris buffer containing trypsin, DNase Ⅰ and RNase A. Control group: fresh annulus fibrosus underwent no treatment. After the de-cellularization process was completed, hematoxylin-eosin (HE) staining was carried out to examine the efficacy on cell removal, and the ultrastructure of annulus fibrosus were observed by scanning electron microscopy. The collagen content, glycosaminoglycan (GAG) content and biomechanical parameters in each group were also detected. Results HE staining and scanning electron microscopy showed that no residual cells were found in Group A, B and C. The structure of annulus fibrosus in Group A was not disturbed, while that in Group B and C was damaged severely and slightly, respectively. There was no statistical difference in collagen content among Group A, B and C, as compared to the control group (P>0.05). But the GAG content was significantly more lower in Group A, B and C than in the control group (P<0.05). There was no statistical difference in ultimate load, ultimate stress, toughness, elastic modulus and mechanical work to fracture between Group A, C and control group (P>0.05), while these parameters of Group B were lower than those in the control group (P<0.05). Conclusions The Triton X-100-treated annulus fibrosus retained the major extracellular matrix composition after cell removal and preserved the major structure and mechanical strength, which is preferable for the construction of tissue engineering annulus fibrosus.