Objective To identify and characterize uveal melanoma associated protein variants with two-dimensional electrophore- sis and mass spectrometry.Design Experiment study.Participants 4 cases of specimens of uveal melanoma and 4 cases of normal con- tributed uveal tissue.Methods Proteins from uveal melanoma and normal urea were separated with two-dimensional eleetrophoresis (2-DE)and visualized with Coomassie G-250.Gels were analyzed by Image Master 5.0 software.The mass spectra were measured by matrix-assisted laser desorption/ionizatian time-of-flight mass spectrometry(MALDI-TOF MS)and searched against NCBI database using Mascot software.Main Outcome Measures Differential proteins.Results A set of 30 proteins were differentially expressed in uveal melanoma compared to nomal urea.Twenty-four types of protein only expressed in uveal melanoma.Five types of protein were up-regu- lated and 1 type of one down-regulated.These proteins can be subdivided into groups according to cellular function,such as enzyme, signal transduction,signal regulation,cytoskehton,immune,etc.Conclusions There is significant difference in protein profilings be- tween uveal melanoma and normal uvea.The differentially expressed proteins may be associated with the development of uveal melanoma.

Objective:To investigate the effect of Astragalus polysaccharides on apoptosis,transdifferentiation and ROS content in renal tubular epithelial cells of diabetic nephropathy.Methods:HK-2 cells were divided into low glucose group,high glucose group and astragalus polysaccharide+high glucose group,cell proliferation was detected by CCK-8 assay after 48 h;cell apoptosis and ROS content was detected by flow cytometry;the expression of E-cadherin,α-SMA,STAT1,STAT3,p-STAT1,p-STAT3 protein were detected by Western blot.Results:The survival rate of cells in high glucose group was significantly lower than low sugar group (P<0.01),cell apoptosis rate,ROS content and E-cadherin,α-SMA,p-STAT1 and p-STAT3 protein expression was significantly higher than low sugar group (P<0.01),the cell survival rate in high glucose+astragalus polysaccharide group was significantly higher than high glucose group,cell apoptosis rate,ROS content and E-cadherin,α-SMA,p-STAT1 and p-STAT3 protein expression was significantly lower than high glucose group (P<0.01).Conclusion:Astragalus polysaccharides can promote the proliferation of renal tubular epithelial cells induced by high glucose,inhibit apoptosis and transdifferentiation,and the mechanism is related to down regulation of JAK/STAT signaling pathway.

OBJECTIVETo explore the expression of connective tissue growth factor (CTGF) in pancreatic cancer and its influence on the proliferation and migration of cancer cells.

METHODSThe expression of CTGF in pancreatic cell line PANC-1 cells was analyzed by real-time PCR and in pancreatic carcinoma (50 cases) tissues by immunohistochemistry. The ability of proliferation and migration in vitro of PANC-1 cells was tested by MTT assay, scratch test and Boyden chamber test after the CTGF gene was overexpressed by Ad5-CTGF or silenced with Ad5-siCTGF transfection.

RESULTSCTGF was overexpressed in both pancreatic cancer cells and tissues. Overxpression of CTGF leads to increased proliferation and migration of PANC-1 cells. The CTGF-transfected PANC-1 cells showed apparent stronger proliferation ability and scratch-repair ability than that of empty vector controls. The results of Boyden chamber test showed that there were 34 cells/field (200× magnificantion) of the CTGF-transfected overexpressing cells, much more than the 11 cells/field of the empty vector control cells; and 6 cells/microscopic field of the Ad5-siCTGF-transfected silenced cells, much less than the 15 cells/field of the control cells.

CONCLUSIONSCTGF is overexpressed in both pancreatic cancer cells in vitro and in vivo, indicating that it may play an important role in the cell proliferation and migration in pancreatic cancer.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Connective Tissue Growth Factor ; genetics ; metabolism ; Humans ; Pancreatic Neoplasms ; metabolism ; pathology ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To observe the effects of hot shock protein 70 (HSP70) inhibitor (PFTμ) on inflammation response in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and mice underwent myocardial ischemia-reperfusion (I/R) injury.Methods RAW264.7 macrophage line of mice was stimulated by LPS as an inflammatory model. These were divided into control (15 min DMSO pretreatment and LPS 2 g/L)and PFTμ treated groups(15 min PFTμ 20 μmol/L pretreatment and LPS 2 g/L). NO concentration was measured by Griess Kit. The expression of iNOS protein and mRNA were detected by Western blot and RT-PCR.Infarct size was determined on mice underwent myocardial ischemia-reperfusion (I/R) injury in the absence or presence (PFTμ 40 mg/kg,intraperitoneal injection).Results PFTμ significantly blocked the production of NO and protein and mRNA expression of iNOS (P<0.05 vs. control). PFTμ also significantly reduced the infarct size on mice underwent I/R injury (P<0.05 vs. control).Conclusion These results suggest that PFTμ could be a potential therapeutic agent for the treatment of inflammatory diseases through inhibiting the production of NO and reducing informatory responses.

OBJECTIVETo understand the effect of pinacidil on rat myocardial Ca(2+)regulation.

METHODSAfter baseline measurement and a period of equilibrium, myocytes were randomly allocated to one of 4 treatment groups: Control group (8 myocytes): incubation in Lactate Ringer's solution at 24 degrees C for 2 hours; K group (8 myocytes): incubation in Lactate Ringer's solution containing 16 mmol/L potassium at 24 degrees C for 2 hours; K+P group (8 myocytes): incubation in Lactate Ringer's solution containing potassium 16 mmol/L and pinacidil 50 micromol/L at 24 degrees C for 2 hours; K+P+G group (8 myocytes): incubation in Lactate Ringer's solution containing potassium 16 mmol/L, pinacidil 50 micromol/L and glibenclamide 10 micromol/L at 24 degrees C for 2 hours. After each incubation, myocytes were resuspended in cell culture media at the same temperature and intracellular [Ca(2+)](i) and SR Ca(2+) release were measured.

RESULTSThe amplitude percent of [Ca(2+)](i) transient evoked by electrical stimulation in the K group was significantly decreased to 67.05% - 80.11% compared to 90.27% - 95.57% in the K+P group during reperfusion after ischemia (P<0.01). The percent amplitude of the [Ca(2+)](i) transient evoked by the rapid application of 10 mmol caffeine in the K group myocyte was approximately 112.00%+/-16.93% compared with that of the [Ca(2+)](i) transient evoked by electrical stimulation. However, in the K+P group myocyte the peak amplitude of the caffeine induced Ca(2+) release was 173.15%+/-26.01% compared with electrical stimulation (P<0.01). The duration of transient evoked by caffeine in K+P group (3.20+/-0.71 ms was significantly shorter than that in K group (3.93+/-0.46) ms (P<0.05).

CONCLUSIONCardioplegic arrest with simultaneous activation of KATP channels preserves rat myocardial Ca2+ by inducing sarcoplasmic reticulum Ca(2+) release and by alteration of Na(+)-Ca(2+) exchanger to better maintain [Ca(2+)](i) homeostasis.

Animals ; Calcium ; metabolism ; Female ; Heart ; drug effects ; Male ; Myocardium ; metabolism ; Pinacidil ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism

This study was purposed to explore the expressions of platelet-activated markers PAC-1 and CD62p in peripheral blood of malignant lymphoma patients and the influence of dipyridamole on their expression. 32 lymphoma patients were divided into simple chemotherapy group (simple group) and chemotherapy plus dipyridamole group (combined group) randomly, and 15 healthy peoples were selected as control group. The dipyridamole of 100 mg/day was given to the patients in combined group. The expression levels of PAC-1, CD62p and fibrinogen (Fib) were detected by flow cytometry and magnetic bead method on day 0, 3, 7 and 14 of chemotherapy respectively. The results showed that the levels of PAC-1, CD62p and Fib in lymphoma patients were significantly higher than those in control group (p < 0.01, 0.05), moreover there was positive correlation between levels of PAC-1 and Fib (r = 0.549, p < 0.01). PAC-1 expression on day 0 and 3 of chemotherapy in simple group was higher than that on day 14 (p < 0.05, 0.01) and CD62p expression on day 3 of chemotherapy was higher than that on day 0, 7 and 14 (p < 0.05, 0.01). PAC-1 expression in combined group on day 14 of chemotherapy was lower than than on day 0 and 3 (p < 0.05, 0.01), and CD62p on day 14 was lower than that on day 3 of chemotherapy (p < 0.05); PAC-1 and CD62p expressions in combined group on day 3, 7 and 14 of chemotherapy were decreased than those in simple group, but Fib level was not changed significantly. It is concluded that the patients with malignant lymphoma usually accompany with platelet activation and hyperfibrinogenemia in peripheral blood. Applying dipyridamole routine dosage in chemotherapy can efficiently restrain platelet activation.
Adolescent ; Adult ; Aged ; Dipyridamole ; therapeutic use ; Dual Specificity Phosphatase 2 ; metabolism ; Female ; Fibrinogen ; metabolism ; Humans ; Lymphoma ; blood ; drug therapy ; Male ; Middle Aged ; P-Selectin ; metabolism ; Platelet Activation ; Young Adult

Human skin cells undergo pathophysiological processes via generation of reactive oxygen species (ROS) upon excessive exposure to ultraviolet B (UVB) radiation. This study investigated the ability of hesperidin (C28H34O15) to prevent apoptosis due to oxidative stress generated through UVB-induced ROS. Hesperidin significantly scavenged ROS generated by UVB radiation, attenuated the oxidation of cellular macromolecules, established mitochondrial membrane polarization, and prevented the release of cytochrome c into the cytosol. Hesperidin downregulated expression of caspase-9, caspase-3, and Bcl-2-associated X protein, and upregulated expression of B-cell lymphoma 2. Hesperidin absorbed wavelengths of light within the UVB range. In summary, hesperidin shielded human keratinocytes from UVB radiation-induced damage and apoptosis via its antioxidant and UVB absorption properties.
Absorption ; Apoptosis* ; bcl-2-Associated X Protein ; Caspase 3 ; Caspase 9 ; Cytochromes c ; Cytosol ; Hesperidin* ; Humans* ; Keratinocytes* ; Lymphoma, B-Cell ; Mitochondrial Membranes ; Oxidative Stress* ; Reactive Oxygen Species ; Skin

OBJECTIVETo investigate the effects of pentoxifylline on rats and mice with learning and memory dysfunctions.

METHODSMorris water maze test was used to observe the effects of pentoxifylline on learning and memory of naturally aging rats, and jumping stand test was performed to examine its effects in promoting the learning and memory functions in mice with scopolamine- and ethanol-induced memory dysfunctions.

RESULTSIn aging rats, pentoxifylline at high, moderate and low doses all significantly reduced the latency of platform finding in the place navigation test (P<0.01 or P<0.05 ), and increased the quadrant searching frequency in the spatial probe test (P<0.05). Pentoxifylline at the 3 doses significantly increased the latency of electrification (P<0.01 or P<0.05) and decreased the times of error (P<0.05) of the mice as compared with scopolamine- treated group. Pentoxifylline also improved ethanol-induced memory dysfunction in the mice, but the changes in the performance of the mice were not statistically significant.

CONCLUSIONPentoxifylline can improve the learning and memory abilities of rats and mice.

Aging ; Animals ; Behavior, Animal ; drug effects ; Ethanol ; Maze Learning ; drug effects ; Memory ; drug effects ; Memory Disorders ; chemically induced ; Mice ; Pentoxifylline ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Scopolamine Hydrobromide

OBJECTIVETo investigate the methods of interventional catheterization for combined congenital heart disease and to evaluate its efficacy in children.

METHODSFrom March 1994 to December 2003, 15 cases (6 boys, 9 girls) underwent transcatheter intervention for combined congenital heart diseases. The procedure of transcatheter intervention was as follows: for pulmonary stenosis (PS) and atrial septal defect (ASD) or patent ductus arteriosus (PDA), PBPV first, occlusion of ASD or PDA later; for coarctation of aorta (COA) and PDA, dilation of COA first, occlusion of PDA 4-15 months later; for aortic stenosis (AS) and PDA, PBAV first, occlusion of PDA later; for ventricular septal defect (VSD) and PDA, all occlusions with detachable coils.

RESULTTranscatheter intervention for combined congenital heart diseases was successful in all patients. There was no residual shunt after occlusion immediately apart from 2 cases of PDA which were little residual after occlusion immediately. Follow-up for (3.57 +/-2.61) years, the systolic pressure gradients across pulmonary valve and coarctation were normal by ultrasonic or transcatheter, except AS. There was 3 cases presented postoperative complications: 1 with mechanical haemolysis, 1 with fall off of coil and 1 with arterial embolism, respectively.

CONCLUSIONTranscatheter intervention for combined congenital heart diseases could obtain satisfactory results with appropriate indications and procedure manipulations.

Abnormalities, Multiple ; surgery ; Cardiac Catheterization ; Catheterization ; Child ; Child, Preschool ; Ductus Arteriosus, Patent ; surgery ; Female ; Follow-Up Studies ; Heart Defects, Congenital ; surgery ; Heart Septal Defects, Atrial ; surgery ; Heart Septal Defects, Ventricular ; surgery ; Humans ; Infant ; Male ; Pulmonary Valve Stenosis ; surgery

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated IC50 value of 3 µM after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G1 phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the PERK/elF2α/CHOP apoptotic pathway, and mitochondrial Ca2+ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER-and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.
Apoptosis ; bcl-2-Associated X Protein ; Caspase 9 ; Cell Death* ; Cell Line ; Colon* ; Colonic Neoplasms* ; DNA Fragmentation ; Endoplasmic Reticulum* ; Extracellular Vesicles ; Humans* ; Inhibitory Concentration 50 ; Lymphoma, B-Cell ; Mitochondria ; Mitochondrial Membranes ; Protein Kinases