10.3969/j.issn.2095-4344.2013.25.004

ObjectiveTo explore the role of ERK1/2 in the expression of the type-1 plasminogen activator inhibitor(PAI-1) induced by parathonnone (PTH) in human renal tubular epithelial cell line HK-2 cells.MethodsVarious concentrentions of PTH and manifold durations were applied in the test.The expression of PAI-1 mRNA and protein in HK-2 cells was measured by RT-PCR and Western blotting,respectively.Besides,ERK1/2 protein was detected by Western blotting before the ERK1/2 inhibitor incubated with the HK-2 cells or after.Results The expression of PAI-1mRNA and protein was gradually up-regulatad along with the increasing concentrations of PTH(10-12-10-10 mol/L).The maximum level of PAI-1 mRNA and protein was detected in 10-10 mol/L PTH and was 4.01 and 3.81 times of control group.Otherwise,the decreased expression of PAI-1 was found while the concentrations of PTH were beyond 10-10 mol/L.The levels of PAI-1 mRNA and protein were increased in pace withtime from 12 to 72 hour,in time-dependent manner,which was 4.06 (12 h) and 4.03 (72 h) times of 0 hour group.The levels of ERK1/2 and PAI-1 were ascended after 10-10 mol/L PTH incubated with the HK-2 cells (all P＜0.01).Howerver,both of them decended after cells were pretreated by the ERK1/2 inhibitor (all P＜0.01),but were still higher than those of control group(all P＜0.05).ConclusionERK1/2 kinase system partly participates in the regulation of PAI-1 induced by PTH in HK-2 cells.

Background Research demonstrated that alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid GluR2 (AMPA-GluR2) is associated with amblyopia.It has been shown that levodopa and cytidine diphosphate choline can improve visual function of amblyopic children,but the mechanism is unclear.Objective This study was to explore the possible effects of levodopa and cytidine diphosphate choline on amblyopia.Methods Monocular deprivation (MD) animal models were created in 60 2-week-old SD rats by monolateral eyelid suturing and observed for 31 days and reared in natural light together with 15 other matched normal healthy SD rats.The models were randomly divided into the MD group,levodopa group,cytidine diphosphate choline group and normal saline control group,with 15 rats for each group.40 mg/kg of levodopa,80 mg/kg of cytidine diphosphate choline,I ml normal saline were given to the rats,respectively,for 28 consecutive days.Expressions of the AMPA-CluR2 protein and AMPA-CluR2 mRNA in the rat visual cortex were detected by immunohistochemistry,Western blot and real-time fluorescence quantitative PCR.Use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The expression values of the AMPA-GluR2 protein (AMPA-GluR2/β-actin) and AMPA-GluR2 mRNA (2-△△Ct) were significantly lower in the MD group than those of the normal control group (protein:0.32 ± 0.02 vs.0.64 ± 0.05,t =13.287,P＜0.05 ;mRNA:0.30±0.01 vs.0.84±0.03,t=38.184,P＜0.05).Those in the levodopa group were significantly increased in comparison with the normal saline solution group (protein:0.59 ±0.04 vs.0.33 ±0.03,t =11.628,P＜0.05 ; mRNA:0.71±0.06 vs.0.33 ±0.02,t =13.435,P＜0.05).The expression values of the AMPA-GluR2 protein and AMPA-GluR2 mRNA were significantly increased in the cytidine diphosphate choline group compared with the normal saline solution group (protein:0.52 ± 0.04 vs.0.33 ± 0.03,t =8.497,P ＜ 0.05 ; mRNA:0.48± 0.04 vs.0.33 ± 0.02,t =7.500,P＜0.05).Conclusions AMPA-GluR2 is associated with the plasticity of visual development.Levodopa and cytidine diphosphate choline may improve visual function by down-regulating the expression of AMPA-GluR2 in the visual cortex.

DNA Fingerprinting ; DNA, Mitochondrial ; Homicide ; Humans ; Mitochondria

Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/diagnosis* ; Death, Sudden/etiology*

Objective:To investigate the frequency of myeloid dendritic cells (mDC) and plasmacytoid dendritic cells (pDC) in peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with renal injury.Methods:The frequency of peripheral mDC and pDC in 102 SLE patients and 10 healthy controls were detected by flow cytometry. The quantitative data were expressed by [ M( P25, P75)]. The measurement data of the two groups with non-normal distribution was analyzed by Mann Whitney U test. The correlation between the two groups was analyzed by Spearman rank correlation analysis and multiple linear regression. Results:The frequency of pDC [14.00%(7.92%, 19.65%) vs 24.55%(19.68%, 32.90%), Z=-3.163, P<0.01] and mDC [21.25%(13.28%, 32.83%) vs 34.85%(24.58%, 41.93%), Z=-2.607, P<0.01] in the peripheral blood of 102 patients with SLE were significantly lower than those of healthy controls. The frequency of pDC [9.09%(7.31%, 17.38%) vs 24.55%(19.68%, 32.90%), Z=-3.033, P=<0.01] and mDC [9.40%(7.88%, 21.60%) vs 34.85%(24.58%, 41.93%), Z=-3.231, P<0.01] in 12 patients with newly diagnosed SLE were also significantly lower than those in healthy controls. After adjustedfor confounding factors, multivariate analysis showed that SLEDAI level was the main factor influencing the frequency of pDC ( P=0.019) and mDC ( P<0.01). In addition, pDC[8.02%(2.25%, 9.97%) vs 16.70%(11.80%, 24.60%), Z=-2.490, P=0.015] and mDC[8.80%(5.99%, 12.80%) vs 20.20%(11.20%, 42.80%), Z=-2.226, P=0.029] in patients with active LN were also significantly lower than that of patients with stable LN. The mDC frequency was positively correlated with the levels of complement C3 ( r=0.455, P<0.01) and C4 ( r= 0.289, P, P<0.01). Conclusion:The frequency of mDC and pDC in SLE patients is significantly abnormal, which is closely related to disease activity. In addition, pDC and mDC may be involved in the occurrence and development of LN.

ObjectiveTo investigate the prevalence of chronic kidney diseases (CKD) from inpatients with cerebrovascular diseases in Shanghai district. MethodsInpatients with cerebrovascular diseases from neurology department of five hospitals in Shanghai from Jun. 2007 to Feb. 2008 were recruited . All the patients were respectively diagnosed by brain CT, CTA, MRI, MRA and TCD. Laboratory data included urinary microalbumin-to-creatinine ratio (ACR), routine urinalysis, fasting plasma glucose, 2-hour-postprandial plasma glucose, Scr, uric acid, etc. All the serum creatinine samples were uniformly tested in central laboratory of Shanghai Ruijin Hospital.Glomendar filtration rate (GFR) was estimated by complicated MDRD equation and CKD stage was classified according to K/DOQI guidelines. ResultsA total of 1014 hospitalized patients with cerebrovascular diseases were enrolled during the observation period, with M/F ratio of 559/455 and mean age of (68.56±12.17) years. Cerebrovascular diseases included ischemic stroke (708 cases), hemorrhagic stroke (197 cases) and transient cerebral ischemie attack (TIA) (109 cases). Microalbuminuria (MAU) was detected in 11.2%, while 24.8% patients had proteinuria. The prevalence of CKD was 47.7%. The percentage of these inpatients in CKD stage 1 to 5 was approximately 6.90%, 14.69%, 21.60%, 2.56% and 1.97% respectively. The Logistic regression model showed that the risk factors of short-term (<30 days) prognosis were albuminuria, hyperglycemia (fast glucose) and anemia. ConclusionsThe prevalence of CKD in inpatients with cerebrovascular diseases was 47.7% in Shanghai. It is significant to evaluate CKD among patients with cerebrovascular diseases, especially to use the screening of ACR in the early stage.

BACKGROUND: Genetic variations of the 5-lipoxygenase activating protein and leukotriene A4 hydrolase genes that confer an increased risk of ischemic stroke have implicated the family of leukotrienes as potential mediators of ischemic stroke. This study aimed to explore the association of ALOX5,LTA4H andLTC4S gene polymorphisms with ischemic stroke risk in a cohort of Chinese in east China.METHODS: This case-control study consisted of 690 patients with ischemic stroke and 690 controls. Polymorphisms ofALOX5 rs2029253 A/G,LTA4H rs6538697 T/C, andLTC4S rs730012 A/C were genotyped by the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. The multivariate logistic regression model was used to exclude the effects of conventional risk factors on ischemic stroke.RESULTS: Carriers of C allele in rs730012 were more susceptible to ischemic stroke (OR: 1.37; 95%CI: 1.08-1.73;P=0.009). The rs2029253 GG genotype showed a risk-reducing effect on ischemic stroke (OR: 0.72; 95%CI: 0.55-0.93;P=0.013) while the rs6538697 CC genotype had an increased risk of ischemic stroke (OR: 1.77; 95%CI: 1.09-2.89;P=0.022). The rs730012 variant was not associated with ischemic stroke risk after adjusting confounding factors (P>0.05).CONCLUSION: The present study suggested that gene polymorphisms in the leukotrienes pathway may exert infl uences, with independent genetic effects, on ischemic stroke susceptibility in a cohort of Chinese in east China.

OBJECTIVETo investigate the feasibility of peptide mass fingerprinting for non-invasive differential diagnosis of IgA nephropathy (IgAN) from the non-IgA nephropathy (IgAN).?

METHODSAccording to the results of renal biopsy, 56 patients were divided into IgAN group (n=28) and non-IgAN group (n=28), and peptide mass fingerprints were acquired from these patients using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).

RESULTSNine different peptides were identified between IgAN and non-IgAN. The two most distinctive differentially expressed peptides, with peptide peak values of 4476.46 and 1968.10, showed area under curve values of 86.18% and 79.77%. Principal component analysis demonstrated that the accumulated explained variance of the first 8 differential peptides reached 95%, suggesting the feasibility of differential diagnosis of IgAN from non-IgAN. Comparison with the Matrix protein database identified the peptide with a relative molecular mass of 5338.08 as a fragment of mucin 4 inform and the 2082.77 peptide as fragment of α1-II type collagen inform.

CONCLUSIONMALDI-TOF MS is feasible for differential diagnosis of IgAN and non-IgAN and also has great potentials in the classification of the subtypes of other systemic diseases.

Adult ; Diagnosis, Differential ; Feasibility Studies ; Female ; Glomerulonephritis, IGA ; diagnosis ; Humans ; Kidney Diseases ; diagnosis ; Male ; Middle Aged ; Peptide Mapping ; methods ; Peptides ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Young Adult

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism