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Objective:RNA interference technology (siRNA) was used to inhibit the expression of DJ-1 gene in lung squamous cell carcinoma HTB-182 cells, then, tandem affinity purification mass spectrometry (TAP-MS) was performed to screen the interacting proteins of DJ-1 in lung cancer cell line of HTB-182.Methods:The siRNA lentivirus vector targeting DJ-1 gene was constructed to infect HTB-182 cells (DJ-1 siRNA group), and the lentivirus vector control group (control siRNA group) and blank control group were established. The expression level of DJ-1 protein was detected by Western blot, and the endogenous DJ-1 protein silenced si-DJ-1-HTB-182 cells were established. The specific primers of DJ-1 were designed, and the DJ-1 expression plasmid pNTAP-DJ-1 with streptomycin binding peptide label (SBP) and calmodulin binding peptide label (CBP) was constructed. The cell line DJ-1 siRNA HTB-182 was stably transfected with liposome, and the positive clones were screened by G418. The positive clones were verified by Western blot, and the interacting proteins of DJ-1 were found by TAP-MS.Results:The protein expression of DJ-1 in DJ-1 siRNA interference group was significantly lower than that in empty plasmid group and blank control group ( P<0.05); HTB182 cell line stably expressing pNTAP-DJ-1 plasmid was successfully constructed; Three proteins interacting with DJ-1 were screened by TAP-MS: cytokeratin 1 (keratin 1), cytokeratin 10 (keratin 10) and NADPH oxidase activating protein P47 (P47 Px). Conclusions:Keratin 1, Keratin l0 and P47 Px protein may be DJ-1 interactions protein.
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Metabolomics, which inherits the ideas of genomics and proteomics, has been widely applied in clinical medical research. In the field of kidney transplantation, changes, which occur in the concentration of small molecule metabolites in blood or urine, can be used to identify organs at risk of acute rejection, locate organ damage, and monitor graft function. This article reviews the problems in the field of kidney transplantation, the concepts, characteristics and research methods of metabolomics, as well as the progress in the application of kidney transplantation and the biomarkers that have been discovered.
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Objective:To explore the effect of trillin on oxidative stress response and nuclear factor E2-related factor 2/antioxidant response element (Nrf2/ARE) pathway in rats after spinal cord injury (SCI). Methods:A total of 108 male Sprague-Dawley rats were randomly divided into sham group (n = 36), model group (n = 36) and trillin group (n = 36), each group was divided into one day, three days and seven days subgroups, with twelve rats in each subgroup. The SCI model was established by modified Allen's heavy strike method in the model group and the trillin group, but no obvious injury in the sham group. The trillin group was given trillin 200 mg/kg every day, and the same amount of normal saline was given in the sham group and model group, twice a day. BBB score was performed one day, three days and seven days after modeling. Morphological changes were tested by Nissl's staining, and the changes of malonaldehyde (MDA) content and superoxide dismutase (SOD) activity were detected by ELISA seven days after modeling. The expression of Nrf2, Kelch like ECH associated protein 1 (Keap1), NAD(P)H quinone oxidoreductase (NQO1) and haemoxygenase 1 (HO-1) were detected by Western blotting one day, three days and seven days after modeling. Results:Compared with the model group, BBB scores increased (P < 0.05); the structure of spinal cord was more complete and the number of Nissl bodies increased; SOD activity increased (P < 0.05) and MDA content decreased (P < 0.05); the expression of Nrf2, Keap1, NQO1 and HO-1 increased (P < 0.05) in the trillin group. Conclusion:Trillin may play a protective role in spinal cord injury by inhibiting oxidative stress response and improving the motor function.
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Objective To explore the role of Galectin3 in transforming growth factor-β(TGF-β)-induced epithelial-mesenchymal transition (EMT) in A549 cells. Methods Galectin3 was over-expressed in an A549 cell line. EMT was induced in lung cancer A549 cells by adding TGF-β. The expressions of Galectin3,E-cadherin,and vimentin were determined by Western blot. The protein expression of E-cadherin and the morphological changes of the cells were detected by immunofluorescence. Cellular proliferation was analyzed with cell counting kit-8,and the cellular migration and invasion was measured by scratches healing and Transwell assay,respectively.Results When only Galectin3 was over-expressed in A549 cell line,the expression levels of EMT-related proteins such as E-cadherin and vimentin were not changed,and the abilities of cellular proliferation,invasion,and migration were not changed either. When the EMT was induced by TGF-β in A549 cells,the E-cadherin expression was down-regulated and the vimentin expression was up-regulated in A549 cells with Galectin3 over-expression. There was no significant change in cellular proliferation,whereas the abilities of cellular invasion and migration were enhanced.Conclusion The TGF-β-induced EMT in A549 cells can be enhanced by Galectin3.
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Objective To investigate the psychological status of organ donation coordinators. Methods The anxiety, depression, sleep quality and personality characteristics of 40 organ donation coordinators were assessed by self-rating anxiety scale (SAS), self-rating depressive scale (SDS), Pittsburgh sleep quality index (PSQI) and Eysenck personality questionnaire (EPQ), respectively. Results The SAS and SDS scores of the organ donation coordinators were significantly higher than those of the norm group (t=23.372, 9.743; both P<0.05). The component scores of sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, use of sleeping medication and daytime function, and the total score of PSQI of the organ donation coordinators were significantly higher than those in the norm group (t=8.054, 7.274, 6.634, 10.613, 8.376, 19.166, 8.496, 17.372; all P<0.05). In terms of EPQ score, the N dimension score of male organ donation coordinators were considerably higher than that of the norm group (P<0.05). No statistical significance was identified in the N dimension score between the female organ donation coordinators and the norm group (P>0.05). The P,E and L dimension scores of the male and female organ donation coordinators did not significantly differ from those of the norm group (all P>0.05). Conclusions Organ donation coordinators present with varying degree of anxiety, depression and poor sleep quality, which deserves sufficient attention. Positive measures should be taken to mitigate these symptoms.
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The basic principles of medical ethics included respect and autonomy,beneficial and nonmaleficence,informed consent and confidentiality,impartiality and mutual assistance.During medical journal editing,medical ethics principles should be strictly applied.Focused on two types of medical thesis,medical scientific research and clinical trials,this paper concluded the ethics issues related to experimental animals,cells and gene researches,all these should be noticed in manuscripts.Meanwhile,this paper analyzed the ethics principles about contrast set and the use of placebo that should be abided in clinical trial,and the respects of patients'privacy during clinical observation,etc.Some review key points were put forward from the perspective of editors.
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To evaluate the clinical effect and safety of western medicine plus Traditional Chinese medicine for sepsis with gastrointestinal dysfunction. We searched CNKI [January 1979 to June 2014], VIP [January 1989 to June 2014], CBM [1978 to 2014], Wan Fang DATA [January 1990 to June 2014], PubMed [1978 to June 2014], The Cochrane Library [Issue 5, 2014], Embase [1974 to June 2014], and other relevant databases and journals to identify randomized controlled trials [RCTs] on western medicine plus Traditional Chinese medicine versus western medicine only for sepsis with gastrointestinal dysfunction. The methodological quality was assessed and the data was extracted according to the Cochrane Reviewer's Handbook and related methods. Meta-analyses were performed by RevMan 5.1.0 software. Five eligible studies included 278 patients. The results of meta-analyses showed that western medicine plus Traditional Chinese medicine therapy can improve the APACHEII score, the peristaltic sound score and SIRS score, improve abdominal distension, decreased white blood cell count, reduce DAO in sepsis patients with gastrointestinal dysfunction. 3 studies reported adverse reactions, there was no significant difference between two groups. Western medicine plus Traditional Chinese medicine can improve gastrointestinal dysfunction in sepsis
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<p><b>INTRODUCTION</b>Diabetes mellitus (DM) is a major cause of chronic kidney disease (CKD). The epidemiology of CKD secondary to type 2 DM (T2DM) (i.e. diabetic nephropathy (DN)) has not been well studied in Singapore, a multi-ethnic Asian population. We aimed to determine the prevalence of CKD in adult patients with T2DM.</p><p><b>MATERIALS AND METHODS</b>We conducted a cross-sectional study on patients (n = 1861) aged 21 to 89 years with T2DM who had attended the DM centre of a single acute care public hospital or a primary care polyclinic between August 2011 and November 2013. Demographic and clinical data were obtained from patients using a standard questionnaire. Spot urine and fasting blood samples were sent to an accredited hospital laboratory for urinary albumin, serum creatinine, HbA1c and lipid measurement. CKD was defined and classified using the 2012 Kidney Disease: Improving Global Outcomes (KDIGO) guidelines and classification.</p><p><b>RESULTS</b>The distribution by risk of adverse CKD outcomes was: low risk, 47%; moderate risk, 27.2%; high risk, 12.8%; and very high risk, 13%. The prevalence of CKD in patients with T2DM was 53%. Variables significantly associated with CKD include neuropathy, blood pressure ≥140/80 mmHg, triglycerides ≥1.7 mmol, body mass index, duration of diabetes, HbA1c ≥8%, age, cardiovascular disease, and proliferative retinopathy.</p><p><b>CONCLUSION</b>CKD was highly prevalent among patients with T2DM in Singapore. Several risk factors for CKD are well recognised and amenable to intervention. Routine rigorous screening for DN and enhanced programme for global risk factors reduction will be critical to stem the tide of DN.</p>
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Cross-Sectional Studies , Diabetes Mellitus, Type 2 , Logistic Models , Prevalence , Renal Insufficiency, Chronic , Diagnosis , Epidemiology , Risk Factors , SingaporeABSTRACT
High expression of Fightless I (FLII) is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins Importin β and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them into the Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.
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Humans , Microfilament Proteins , Metabolism , Nuclear Pore Complex Proteins , Metabolism , Receptors, Cytoplasmic and Nuclear , Metabolism , Recombinant Fusion Proteins , Metabolism , beta Karyopherins , MetabolismABSTRACT
Objective To investigate the influence of forceps delivery or emergency cesarean section on the maternal and infant outcome in pregnant women with abnormal second stage of labor.Methods We retrospectively analyzed the clinical data of 118 parturients with abnormal second stage of labor in the department of obstetrics and gynecology in our hospital from 2013 January to 2015 February.The involved parturients with abnormal second stage of labor received different delivery modes including forceps delivery and emergency cesarean section, and the delivery time and its influence on the maternal and infant outcome were compared.Results The mean delivery time of forceps delivery and emergency cesarean section was (14.8 ± 5.3) and (32.7 ± 12.6) minutes, the difference was significant (P＜0.05).The bleeding probability and the occurrence of severe neonatal asphyxia in the forceps delivery group was obviously lower than that of maternal emergency cesarean section group (P＜0.05).The influence of emergency cesarean section on the newborn severe asphyxia was more serious than forceps delivery (P ＜0.05).There was no statistical difference in the mild asphyxia, facial bruising, scalp hematoma and neonatal pneumonia between the two groups.Conclusion Compared with emergency cesarean section, forceps delivery can protect the safety of mother and child can reduce dystocia risk coefficient for the pregnant women with abnormal second stage of labor.
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<p><b>OBJECTIVE</b>To investigate whether the cardioprotective effect of hemin against ischemia/reperfusion (I/R) injury is through the inhibition of calpain activity, and to explore its underlying mechanism.</p><p><b>METHODS</b>Sixty-four SD rats were randomly divided into eight groups (n = 8): sham, I/R, MDL+ I/R, MDL, hemin + I/R, hemin, and ZnPP + hemin+ I/R, ZnPP. Iangendorff isolated rat heart perfusion model was used. The rat hearts were suffered from 40 min of ischemia followed by 30 min of reperfusion. After that, left ventricular developed pressure (LVDP) was recorded. Infarct size and release of lactate dehydrogenase (LDH) were measured. Calpain, heme oxygenase (HO), and caspase 3 activities were evaluated. Expression of calpastatin protein was detected by Western blot.</p><p><b>RESULTS</b>(1) After suffered from ischemia/reperfusion, the calpain activity and caspase 3 activity increased. MDL28170, an inhibitor of calpain, prevented ischemia/reperfusion induced increases in LDH and infarct size, improved the LVDP recovery. (2) Compared with ischema/reperfusion rat hearts, pretreatment of hemin enhanced the HO-1 activity, decreased the calpain and caspase 3 activities, declined LDH release and infarct size, and improved LVDP recovery. (3) Ischemia/reperfusion reduced the expression of calpastatin protein in rat hearts, which was inhibited by hemin pretreatment. And HO-1 inhibitor could abolish the cardioprotection of hemin.</p><p><b>CONCLUSION</b>Cardioprotective effect of hemin against ischemia/reperfusion injury is through the inhibition of calpain activity, the mechanism might be involved in the increase in calpastatin protein expression.</p>
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Animals , Rats , Calpain , Metabolism , Cardiotonic Agents , Pharmacology , Caspase 3 , Metabolism , Heme Oxygenase-1 , Metabolism , Hemin , Pharmacology , L-Lactate Dehydrogenase , Metabolism , Myocardial Reperfusion Injury , Drug Therapy , Rats, Sprague-DawleyABSTRACT
Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic (37-38°C) ischemia, mild hypothermic (31-32°C) ischemia, hyperthermic (41-42°C) ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20-min ischemia. The expression of c-fos protein and the levels of malondialdehyde (MDA) and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-fos protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion following 20-min ischemia as compared with the sham-operated group (P<0.01). The levels of MDA and lactate in mild hypothermic group were significantly lower than those in normothermic group (P<0.01). It is suggested that brain temperature influences the translation of the immunoreactive protein product of c-fos after global cerebral ischemia, and MDA and lactate are also affected by hypothermia and hyperthermia.
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Animals , Male , Rats , Body Temperature , Brain , Metabolism , Brain Ischemia , Metabolism , Cerebral Cortex , Metabolism , Hippocampus , Metabolism , Immunochemistry , Lactic Acid , Metabolism , Malondialdehyde , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Proto-Oncogene Proteins c-fos , Metabolism , Rats, Sprague-Dawley , Reperfusion Injury , Metabolism , Spectrophotometry , Temperature , Time Factors , Tumor Suppressor Protein p53 , MetabolismABSTRACT
OBJECTIVE@#To investigate the expression pattern of adapter protein with a Src-homology 2 domain (SH2B1), the suppressor of cytokine signaling-3 (SOCS3), protein-tyrosine phosphatase 1B (PTP1B) and neturopetide Y (NPY) in obese and normal mice hypothalamus and its relation with serum leptin and insulin levels.@*METHODS@#The obesity animal model was prepared with healthy C57/bl6 mice. Lee's index and Homeostasis model assessment-insulin resistance (HOMA-IR) were calculated. The mRNA levels of SH2B1, SOCS3, PTP1B and NPY were measured by fluorescent quantitation RT-PCR. The SH2B1 and NPY protein expressions were detected by Western blot.@*RESULTS@#Compared with the normal mice of the same age, SH2B1 mRNA expression in the obese mice hypothalamus decreased. SOCS3 and PTP1B mRNA expression increased. Western blot showed that SH2B1 protein expression decreased, while NPY protein expression increased in the obese mice. Linear correlation analysis showed that the serum leptin and fasting insulin levels were negatively correlated with SH2B1mRNA expression and positively correlated with SOCS3 and PTP1B mRNA expression.@*CONCLUSION@#SH2B1, SOCS3, PTP1B and NPY are key factors for obesity development.
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Animals , Mice , Adaptor Proteins, Signal Transducing , Metabolism , Hypothalamus , Metabolism , Insulin , Blood , Insulin Resistance , Leptin , Blood , Mice, Inbred C57BL , Neuropeptide Y , Metabolism , Obesity , Metabolism , Protein Tyrosine Phosphatase, Non-Receptor Type 1 , Metabolism , RNA, Messenger , Suppressor of Cytokine Signaling 3 Protein , Suppressor of Cytokine Signaling Proteins , MetabolismABSTRACT
Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic (37-38°C) ischemia, mild hypothermic (31-32°C) ischemia, hyperthermic (41-42°C) ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20-min ischemia. The expression of c-fos protein and the levels of malondialdehyde (MDA) and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-fos protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion following 20-min ischemia as compared with the sham-operated group (P<0.01). The levels of MDA and lactate in mild hypothermic group were significantly lower than those in normothermic group (P<0.01). It is suggested that brain temperature influences the translation of the immunoreactive protein product of c-fos after global cerebral ischemia, and MDA and lactate are also affected by hypothermia and hyperthermia.
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OBJECTIVE@#RNA interference technology (siRNA) was used to inhibit the expression of DJ-1 gene in lung squamous cell carcinoma SK-MES-1 cells, and the cell biological behaviors were investigated to explore the function of DJ-1 gene.@*METHODS@#A targeted DJ-1 siRNA lentiviral vector with a green fluorescent protein (GFP) as a reporter was constructed. The constructed DJ-1 siRNA and control-siRNA vectors were infected into SK-MES-1 cells as experimental (DJ-1 siRNA) and control (Control siRNA) groups, respectively. The DJ-1 protein expression was determined by Western blot. The cell proliferation capability was measured with methyl thiazolyl tetrazolium (MTT). The cell cycle was analyzed by flow cytometry. The capability of cell migration was determined by Transwell method.@*RESULTS@#Compared with control-siRNA and blank-control groups, the protein expression of DJ-1 gene was down-regulated, the capability of cell proliferation was obviously inhibited (P<0.01), the cell cycle was arrested with increased number of G1- and G2-phase cells and reduced number of S-phase cells, and the capability of cell migration was significantly decreased (P<0.01) in the DJ-1 siRNA-infected cells.@*CONCLUSION@#DJ-1 gene might play a role in promoting cell proliferation and cell migration capability in vitro in lung cancer SK-MES-1 cells.
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Humans , Base Sequence , Carcinoma, Squamous Cell , Genetics , Pathology , Cell Line, Tumor , Cell Movement , Genetics , Cell Proliferation , Genetic Vectors , Genetics , Intracellular Signaling Peptides and Proteins , Genetics , Metabolism , Lentivirus , Genetics , Metabolism , Lung Neoplasms , Genetics , Pathology , Molecular Sequence Data , Oncogene Proteins , Genetics , Metabolism , Protein Deglycase DJ-1 , RNA Interference , RNA, Small Interfering , GeneticsABSTRACT
OBJECTIVE@#To determine the effect of RNA interference with transferred pshRNA/PHB on the biological characteristics of paclitaxel-resistant ovarian cancer cell lines.@*METHODS@#Western blot and real time-PCR were used to assay the expression of PHB protein and mRNA in SKOV3/Taxol-25 and SKOV3 cell lines. The SKOV3/Taxol-25 cell lines were transiently transfected by 3 target-specific small hairpin RNA (shRNA) interference fragments with fluorescent protein named the pshRNA427/PHB1, pshRNA248/PHB2, and pshRNA136/PHB3. The empty plasmid transfection via vehicle Lipofectamine2000 served as a negative control. The expression levels of PHB protein and mRNA were detected by Western blot and real time-PCR after the transfection for 48 h. The silence effect of PHB1 and PHB3 groups was obvious. PHB1, PHB3, and the negative control groups were used for the following experiments. MTT and flow cytometry assay were used to test the cell proliferation, IC50 of paclitaxel, and cell apoptosis in the 3 groups.@*RESULTS@#The expression levels of PHB protein and mRNA (2(-ΔΔCt)) were significantly higher in SKOV3/Taxol-25 cell line than those in SKOV3 cell line (P<0.05). The expression levels of PHB protein and mRNA were significantly lower in the PHB1 and PHB3 groups than those in the negative control group (P<0.05). The cell proliferations in the PHB1 and PHB3 groups were obviously slower than those in the negative control group after transfection for 48 h and 72 h (P<0.05). The IC50 of paclitaxel in the PHB1 and PHB3 groups significantly decreased after transfection for 72 h compared with the negative control group(P<0.05). The cell apoptotic rate in the PHB1 and PHB3 groups significantly increased after transfection for 48 h compared with the negative control group (P<0.05).@*CONCLUSION@#The shRNA/PHB can effectively suppress the expression of PHB gene in paclitaxel-resistant ovarian cancer cell lines. The cell proliferation in paclitaxel-resistant cell lines with removed PHB gene is significantly reduced. The apoptotic rate and the paclitaxel sensitivity of resistant cell lines with removed PHB gene are significantly increased. PHB gene is related to paclitaxel-resistance and interfering PHB gene expression may reduce paclitaxel resistance in ovarian cancer.
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Female , Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Drug Resistance, Neoplasm , Genetics , Ovarian Neoplasms , Genetics , Metabolism , Pathology , Paclitaxel , Pharmacology , RNA Interference , RNA, Messenger , Genetics , Metabolism , RNA, Small Interfering , Genetics , Repressor Proteins , Genetics , Metabolism , Transfection , Tumor Cells, CulturedABSTRACT
OBJECTIVE: To stereospecifically synthesize disaccharide galactopyranosyl(α1 → 2) glucopyranoside with protecting groups which are easily and selectively deprotected. METHODS: With D-glucose as a raw material, a donor methyl 3, 4, 6-tri-O-benzoyl-β-D-glucopyranoside was synthesized through acetylation, bromination, methylation, deacetylation, benzylidenation, selective benzoylation, chloroacetylation, and dechloroacetylation; the glucopyranosyl acceptor and a donor isopropyl 3-O-allyl-2-O-benzoyl-4, 6-O-benzylidene-β-D-1-thiogalactopyranoside were reacted at catalysis to achieve an α-coupled product. RESULTS: At the catalysis of TMSOTf/NIS, galactopyranosyl donor and glucopyranosyl acceptor were coupled to afford stereospecifically a full protected disaccharide galactopyranosyl(α1 → 2) glucopyranoside fragment, methyl 3-O-allyl-2-O-benzoyl-4, 6-O-benzylidene-α-D-galactopyranosyl(1 → 2)-3, 4, 6-tri-O-benzoyl-β-D-glucopyranoside. All products were characterized with NMR and MS etc. CONCLUSION: A full protected 1, 2-cis disaccharide fragment was stereospecifically obtained through coupling reaction due to concerted effect of the special configurations of the galactopyranosyl donor and the glucopyranosyl acceptor. Copyright 2012 by the Chinese Pharmaceutical Association.
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<p><b>BACKGROUND</b>The optimal time window for the administration of hypothermia following cerebral ischemia has been studied for decades, with disparity outcomes. In this study, the efficacy of mild brain hypothermia beginning at different time intervals on brain endogenous antioxidant enzyme and energy metabolites was investigated in a model of global cerebral ischemia.</p><p><b>METHODS</b>Forty-eight male Sprague-Dawley rats were divided into a sham-operated group, a normothermia (37°C - 38°C) ischemic group and a mild hypothermic (31°C - 32°C) ischemia groups. Rats in the last group were subdivided into four groups: 240 minutes of hypothermia, 30 minutes of normothermia plus 210 minutes of hypothermia, 60 minutes of normothermia plus 180 minutes of hypothermia and 90 minutes of normothermia plus 150 minutes of hypothermia (n = 8). Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model for 20 minutes and mild hypothermia was applied after 20 minutes of ischemia. Brain tissue was collected following 20 minutes of cerebral ischemia and 240 minutes of reperfusion, and used to measure the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and adenosine triphosphate (ATP).</p><p><b>RESULTS</b>Mild hypothermia that was started within 0 to 60 minutes delayed the consumption of SOD, GSH-Px, GSH, and ATP (P < 0.05 or P < 0.01) in ischemic tissue, as compared to a normothermic ischemia group. In contrast, mild hypothermia beginning at 90 minutes had little effect on the levels of SOD, GSH-Px, GSH, and ATP (P > 0.05).</p><p><b>CONCLUSIONS</b>Postischemic mild brain hypothermia can significantly delay the consumption of endogenous antioxidant enzymes and energy metabolites, which are critical to the process of cerebral protection by mild hypothermia. These results show that mild hypothermia limits ischemic injury if started within 60 minutes, but loses its protective effects when delayed until 90 minutes following cerebral ischemia.</p>
Subject(s)
Animals , Male , Rats , Adenosine Triphosphate , Metabolism , Antioxidants , Metabolism , Brain Ischemia , Metabolism , Glutathione , Metabolism , Glutathione Peroxidase , Metabolism , Hypothermia, Induced , Rats, Sprague-Dawley , Superoxide Dismutase , Metabolism , TemperatureABSTRACT
Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.
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OBJECTIVE@#To establish the protein expression map of nasopharyngeal carcinoma (NPC), and provide a basis for proteomic study of NPC.@*METHODS@#Laser capture microdissection (LCM) was used to isolate cancer cells from NPC tissues. Two-dimensional gel electrophoresis(2-DE) was used to separate the total proteins of LCM purified NPC cells. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) was performed to identify the proteins, and bioinformatics was used to construct the 2-DE database of NPC proteome.@*RESULTS@#A 2-DE reference map of NPC was established. On the 2-DE map, a total of (1 312+/-30) protein spots were detected, and 427 protein spots representing 241 non-redundant proteins were identified. The 2-DE database of NPC proteome was constructed. These data could be accessed at our website (http://www.xyproteomics.org).@*CONCLUSION@#A protein expression profile of LCM purified NPC tissues has been established for the first time, which provides useful information and source for proteomic study of NPC.