Objective To establish and optimize the two-dimensional gel electrophoresis technical platform for the blood serum proteome research in patients with end stage renal disease(ESRD).Methods Immobiline pH gradients isoelectric focusing was used as the first dimensional gel electrophoresis and the vertical SDS-PAGE was used as the second dimensional gel electrophoresis(2-DE).The 4 differentially expressed protein spots were identified by mass spectrometry.Results Satisfactory 2-DE maps of ESRD patients serum protein were obtained and there were some differentially protein spots between the 2-DE maps of ESRD patients and normal controls.Conclusions The 2-DE technology for the serum proteome of ESRD patients is set up.

OBJECTIVE: To develop a method for the determination of levodopa in human plasma and study aarodopa's pharmacokinetics. METHODS: Experiments were performed on Waters 2010 HIPLC system with a 474 fluorecence detector (?EX = 278nm, ?Em = 325nm) and a Lichrospher 100 C18 column. The mobile phase(pH3. 7) was 90% water containing 0.08 mmol/L EDTA, 70mmol/L KH2PO4, 2. 08mmol/L sodium heptanesulfonate and 10% methanal. The flow rate was 1. 0ml/min. The plasma sample was directly injected for determination after being deproteinized with perchloric acid. RESULTS: The linear range was 0. 125-4.0ug/ml,the limitation of detection was 0. 25ug/ml,within- day RSD

Objective:Our aim is to investigate the relationship of VEGF-C,VEGF-D and lymph node metastasis in pancreatic cancer,and to elucidate the role and significance of the cancerous peripheral lymphatic tissue.Methods: The expression of VEGF-C and VEGF-D was assayed by means of immunohistochemistry in 30 pancreatic carcinomas.Results:The positive rates of VEGF-C、VEGF-D were 73% (22/30)、57% (17/30) respectively in pancreatic cancer.The expression of VEGF-C、 VEGF-D in cancerous invasive edge was significantly higher than that in the center of cancerous tissues.There was no correlation between the expression of VEGF-C、VEGF-Dand the site,differentiation,histology types.Ⅲ~Ⅳ stages of pancreatic cancer showed strong expressions of VEGF-C、VEGF-D than that Ⅰ~Ⅱ stages of pancreatic cancer.The positive lymph node in positive VEGF-C and VEGF-D group were higher than that in the negative group.Conclusion:VEGF-C and VEGF-D induced lymphangiogenisis in pancreatic cancer,promoted the tumor cell lymph metastasis.

To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.
Fibrosis ; Kidney/*pathology ; Kidney Diseases/etiology ; Kidney Diseases/pathology ; Kidney Diseases/*prevention & control ; Mycophenolic Acid/*analogs & derivatives ; Mycophenolic Acid/*pharmacology ; Random Allocation ; Rats, Sprague-Dawley ; Ureteral Obstruction/*complications

The different sera proteomic components between uremia patients and normal subjects were studied through two-dimensional gel electrophoresis technique. Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2DE), silver staining, ImageMaster 2D 5.0 analysis software, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS) and IPI human database searching were used to separate and identify the proteome of the sera from the patients with uremia. The results showed that satisfactory 2DE patterns of the serum proteins were obtained. Twenty-six protein spots showed significant difference in quantity in uremia patients, and 20 protein spots were identified by MALDI-TOF-TOF-MS. It was concluded that good reproducibility could be obtained by applying immobilized pH gradient 2DE to separate and identify the proteome in serum, which provided the foundation for the further study on uremia toxins pertaining to protein.
Blood Protein Electrophoresis ; Blood Proteins/*analysis ; Case-Control Studies ; *Electrophoresis, Gel, Two-Dimensional/methods ; Proteome/*analysis ; Proteome/chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/*methods ; Uremia/*blood

Objective To investigate the fungal infection situation and the risk factors in lupus nephritis(LN)patients.Methods Infection states were investigated in 93 LN patients and 45episodes(about 48.4%) occurred the different kind and different degree of Hospital-acquired infections.The fungal infection rate was 22.6% and two patients directly died of it.Except fungi,the main pathogens were bacteria and virus.The main infection sites were urinary,gastrointestinal and respiratory tract.Results The infection causes were related to the use of corticosteroids,immunodepressive,antibiotic and the aggressive operation,renal failure ,the duration in hospital and so on.Conclusion Effective prevention and treatment for infection in LN patients is an important way to improve the prognosis of LN.

AIM: To investigate the effect of high glucose concentration on serum and glucocorticoid induced protein kinase (SGK) mRNA and protein expressions in human proximal tubular epithelial cells (HKC) and the possible role of SGK in the production of extracellular matrix (ECM) of HKC under the condition of high glucose. METHODS: HKC was divided into 3 groups: control glucose group (CG group, 5 5 mmol/L D-glucose); high glucose group (HG group, 25 mmol/L D-glucose) and osmotic control group (MG group, 19 5 mmol/L mannitol and 5 5 mmol/L D-glucose). The expressions of SGK mRNA and protein were assessed by semi-quantitative RT-PCR and Western blotting respectively. The level of secretary and cytoplasmic fibronectin (FN) were detected by enzyme-linked immunoabsordent assay (ELISA) and indirect-immunofluorescence. RESULTS: HKC expressed SGK1, SGK2 and SGK3 at mRNA and protein levels. Their mRNA level were up-regulated since 2 hours after cells exposed to D-glucose and this up-regulation persisted to the end of 8th hour, and SGK1 protein level elevated simultaneously. On the other hand, the increased FN secretion by high glucose was in a time-dependent manner and its improved secretion threshold was just followed by the high expression of SGK1. CONCLUSIONS: In response to high glucose, the expression of SGK1, SGK2 and SGK3 in human proximal tubular epithelial cells were up-regulated which was accompanied with FN accumulation. The high expression of SGK may mediate overproduction of ECM in proximal tubular epithelial cells and contribute to the diabetic nephropathy.

To investigate the expression and the role of three isoforms of Serum and Glucocorticoid-inducible Kinase (SGK) in experimental diabetic nephropathy (DN), 12 male C57BL/6 mice of 8-weeks-old were divided into two groups. Streptozotocin (STZ)-induced diabetic nephropathy and normal controls were analyzed at the end of the 4th week after the induction of diabetes. Renal hemodynamics and histological studies were performed. The expression of SGK1 mRNA, SGK2 mRNA and SGK3 mRNA of kidney cortex were measured by RT-PCR, and the cortical SGK1 protein was detected with Western blotting. Our results showed that the blood glucose, blood HbA1c, 24h urinary protein, creatinine clearance and the renal index were all increased in DN group. More extracellular matrix (ECM) accumulation was observed. The level of cortical SGK1 mRNA and protein were up-regulated in DN group in comparison with control group. SGK2 and SGK3 mRNA were elevated in DN mice. In DN, mRNA level of three SGK isoforms and SGK1 protein were increased significantly. It is concluded that SGKs may contribute to the early renal injury of DN.

Objective To evaluate the mechanical homogeneity pattern of isolated carotid atherosclerotic plaque intima on longitudinal axis view using ultrasonic velocity vector imaging.Methods Fourty six patients with 48 isolated plaques were undergone the high frequency ultrasound scanning,the real-time two-dimensional longitudinal-axis gray scale view at the maximal thickness of the plaque was obtained in three complete cardiac cycles.A dedicated velocity vector imaging(ⅤⅤⅠ)workstation was used for the off-line dynamic two-dimensional gray-scale image analysis and mechanics parameter(i.e.,the peak strain and the absolute difference of the peak strain)assessment at the sites of the upper,middle and down stream separately.The length and direction distribution of the velocity vectors at the carotid atherosclerotic plaque intima were observed.Wilcoxon test was utilized for the comparison of the peak strain and the absolute difference of the peak strain between two sites.Results The chaos phenomenon of velocity vector distribution at the sites of plaque intima occurred significantly differed from those uniform velocity vectors at the reference sites.According to the two groups of the higher strain value side and the lower strain value side on the upper stream and the down stream of the plaque intima,the peak strain at the higher strain value side was significantly higher than the lower strain value side(P=0.000).There was a significantly difference between the absolute difference of peak strain from the higher strain value side to the middle stream point of the plaque intima and the absolute difference of peak strain from the lower strain value side to the higher strain value side of the plaque intima(P＜0.017).Conclusions The length and direction distribution of velocity vectors of the isolated carotid atherosclerotic plaque intima on longitudinal axis view are uneven,the mechanical heterogeneity pattern exists at the plaque intima and could be visualized and quantitatively evaluated using ultrasonic velocity vector imaging.

Objective To understand the histological characteristics of the major endocrine organs of tree shrew , and provide a normal histological atlas of endocrine organs of tree shrew .Methods Ten artificially fed healthy tree shrews were killed and dissected after anesthesia .The thyroid, parathyroid, adrenal and pituitary glands were observed by gross inspection and samples were taken for routine histological examination with HE staining .Results ( 1 ) The thyroid gland was pale yellow, located on both sides of the 2-4 tracheal rings.The thyroid gland was plate-shaped, its surface was covered with a thin fibrous capsule . The thyroid parenchyma was divided into several lobules by stretched capsule membrane .Follicular and parafollicular cells were distributed in the lobules , and red colloid was present in follicular cavity.(2) Each side had one parathyroid , located on the cranial or the outer surface of the middle part of the thyroid gland, and was slightly covered by thyroid .The gland was round or oval , and its parenchyma was made up of the principal cells and eosinophil cells , and acinar structure appeared in the parenchyma .( 3 ) The adrenal glands were oval , yellow color, located in the renal hili , and linked to the kidneys .They were surrounded by a thin capsule .The parenchyma was divided into cortex and medulla .The cortex was divided into zona glomerulosa , zona fasciculata and zona reticularis from outside to inside.The zona glomerulosa was the thickest layer and the zona fasciculata was the thinnest .The medulla cells formed clumps or mesh, with central vein in the central part .(4) The pituitary gland was located in the sella turcica , with no recessus hypophysis .The pituitary gland was composed of the adenohypophysis and neurohypophysis .Its surface was covered with a connective tissue capsule .The pituitary gland was divided into distal part , middle part and pars tuberalis . neurohypophysis was made up of neural and pars infundibularis .Conclusions The histological atlas of endocrine organs in the tree shrew is established , which is close to that of the primate animals in the morphology , and provide histological evidence for the study of tree shrew endocrine organs and disorders , as well as the animal model of human diseases .