ObjectiveThis study explored the change of IL-18,and IFN-γ and their role in the pathogenesis of primary Sj(o)gren's syndrome (pSS) by determining the expression of IL-18,IFN-γ in labial glands and blood of patients with pSS.MethodsThe levels of IL-18,IFN-γ in labial glands of patients with pSS were determined by immunohistochemical method.The level of serum IL-18 was determined with enzyme linked immunosorbent assay ELISA.The level of γ globulin was measured by the cellulose acetate membrane method.SPSS 17.0 statistical software was used for data analysis.The t test,Fisher's exact probability test and index correlation analysis were performed with linear correlation analysis.Spearman's test was used to examine the association between the expression of IL-18 and IFN.ResultsThe positive rate of IL-18 and IFN-γ in patients with pSS was much higher than normal control group(P＜0.05).And the IL-18 and IFN-γ levels were positively correlated withpatients with pSS (P＜0.05).The level of serum IL-18 in patients with pSS was (216±85) pg/ml,which was much higher than normal control group(39±8) pg/ml.The IL-18 level and γ globulin(23.2±2.6)％ was positively correlated in patients with pSS (r=0.538,P＜0.05).ConclusionThe expression of IL-18 and IFN-γ in labial glands of patients with pSS are much higher in patients with pSS than those of the normal controls.Theresults have shown that IL-18 and IFN-γ may be involved inthe pathogenesis of pSS.

Objective To study glucose metabolism in patients with liver cirrhosis and to explore the relationship between glucose and liver function.Methods 164 liver cirrhosis patients with abnormal glucose metabolism were divided into A,B and C groups according to Child-Turcotte-Pugh(CTP)classification system.Glucose metabolic disorder and relation between blood sugar level and liver function were observed and analyzed.Results We divided the patients into three subgroups according to their blood sugar levels:hypoglycemia group,impaired fasting glucose group and diabetes mellitus group.Patients of Grade C were with the highest incidence of hypoglycemia,with a P value P

Circulating nitric oxide(NO)and endothelin (ET) levels were determined in 30 normal subjects and 63 diabetic patients.The results showed that:serum NO level was significantly higher in DM I group(68.66?12.37?mol/L)and DM Ⅱ group(63.43?11.09?mol/L)than in normal subjects(44. 92?9.04?mol/L,P0.05).Plasma ET level was markedly elevated in DM Ⅰ group(68.92?11.96ng/L,P

Objective To investigate se rum concentration of tumor necrosis factor(TNF)-? and its correlation with insulin resistance in pregnant women with gestational diabetes mellitus (GDM). Methods Enzyme-linked immunosorbent assay was used to measure th e fasting serum TNF-? levels of 42 women with GDM (28～39 gestational weeks), a n d 40 cases of normal pregnant women in the third trimester. Fasting plasma gluc ose, insulin, C-peptide and glycosylated hemoglobin (HbA1c) were also measured at the same time. Insulin sensitive index (ISI) was calculated. Res u lts (1)Significantly elevated serum TNF-? was found in the women w i th GDM(5.2?1.6) ng/L as compared with the healthy pregnant women in third t rimester (4.5?0.5)ng/L(P

Objective To observe the phenotypic changes between renal tubular epithelial cells and mesenchymal cells in rat tubulointerstitial fibrosis(TIF) .Methods The renal tubulointerstitial fibrosis models of Wistar rats were established by gavage with excessive adenine. The morphological changes of model kidney were observed by light microscopy, electron microscopy, polarizing microscopy. The?-smooth muscle actin(a-SMA), vimentin, and keratin were examined by immunohistochemistry staining. While the proliferating cell nuclear antigen (PCNA) was also assessed. Results There were many 2, 8-dioxyadenine crystals derived from the metabolite of adenine to deposit in the renal tubules, which induced the injury. Tubular epithelial cells degeneration and regeneration, loss of renal tubules, mononuclear cells infiltration in renal interstitium and the accumulation of extracellular matrix protein were observed. Both ?-SMA and vimentin were positive in some of renal tubules. And consecutive sections of immunohistochemical staining showed that the area of distribution of a-SMA positive cells was almost the same area as that of vimentin positive cells. Polarizing microscopy results showed that there was a mass of interstitial collagen ( Ⅲ and Ⅰ) accumulation. And electron microscopy examination proved that epithelial cells invaded into the renal interstitium. Conclusion There is tubular epithelial cells-myofibroblasts transdifferentiation in the process of TIF, and which plays a key role in the accumulation of extracellular matrix.

Objective To study the effects of high albumin, high glucose and low bovine serum on the phenotypic transformation of renal tubular epithelial cells. Methods The normal human kidney proximal tubular eel] line (HKC) was cultured for 30 days in the presence of high albumin (1.5 g/L), high glucose(25 mmol/L) and low bovine serum(2% ) . Morphological changes were observed by electronic microscopy. Immunohistochemistry stain was used to examine the expression of cytokeratin, vimentin, a-SMA, collagen Ⅰ and TGF-pl protein. Western blot was applied to further detect the process of collagen I protein expression, and in situ hybridization was used to examined the expression of collagen Ⅰ gene. Results Renal tubular epithelial cells cultured in high albumin, high glucose and low bovine serum showed obvious morphologic changes, including elongated shape, decrease of microvilli and mitochondria, and increase of rough endoplasmic reticulum under electronic microscopy. Immunohistochemistry stain revealed the reduction of cytokeratin, and enhancement of vimentin, ?-SMA, TGF-?1 and collagen Ⅰ. Western blot demonstrated that the expression of collagen Ⅰincreased in a time-dependent manner, and in situ hybridization showed that collagen type Ⅰ mRNA increased as well. Conclusion High albumin, high glucose and low bovine serum induce phenotypic transformation of renal tubular epithelial cells into mesanchymal cells.

In order to adapt to the demand of culturing high quality talents and to strengthen student's comprehensive quality,pathophysiology department of medical college of Qinghai University carried on the reform on the pathophysiological experiment examination system.After researches and practices in recent years,we established a set of relatively sound examination system including experiment operating and experiment report in normal study,report for designable experiment and experiment skill and theory exam.Investigation results demonstrated that examination system after reform functioned well in training the students' operation ability,observation ability and ability to analyze and solve questions.

AIM:To provide evidence for the molecular mechanism of homocysteine (Hcy) as an independent risk factor in atherosclerosis (AS) by investigating the effect of Hcy on phenotype transformation and proliferation of vascular smooth muscle cells(VSMCs) in rats.METHODS:After treated with different concentrations of Hcy for 24 h,the cultured VSMCs were assayed for cell proliferation rate by MTT method,cell cycle by flow cytometry,the expression of SM22-? mRNA by semi-quantitative RT-PCR and the observation of morphological characteristics and the phenotype transformation by transmission electron microscopy.RESULTS:Hcy increased the cell proliferation rate and gradually reduced the proportion of the cells in G0 /G1 phase.Hcy down-regulated the expression of SM22? mRNA and the most significant effect was observed at concentration of 1 000 ?mol/L.The observations of transmission electron microscopy revealed an abundant endoplasmic reticulum,Golgi's complex,loose nucleus and puffy chromatin in VSMCs treated with high concentration of Hcy.CONCLUSION:Hcy promotes the proliferation and phenotype transformation of VSMCs simultaneously.

Objective: To study the axonal effect and the expression of integrin α6β4 during Schwann cell(SC) differentiation and myelination. Methods: Schwann cells were dissociated from the sciatic nerve of neonatal Waster rats and neurons dissociated from spinal cord. Singal cultures and purified populations of SC were cocultured with NC. Four methods (contrast microscope, scanning electron microscopy(SEM), immunocytochemistry method and in situ hybridization ) were used. Results: The separately cultured Schwann cells showed MBP negetive by immunocytochemistry method. But cocultured SC were shown positive. SEM showed that Schwann cells' membrane loop progressively circumnavigated around the axon during myelination, which suggested that the non-myelinating SC(nMSC) transformed to myelinating SC (MSC). In situ hybridization showed integrin α6β4 positive signals only on the outer surface of the Schwann cell-axon unit in SC coculture with NC. Conclusion: The differentiation and maturation of SC depend on axon, and the activity of integrins is expressed by axon. Axonal contact induces the expression of α6β4 during SC myelination, which suggests that integrin α6β4 is an important mediator of interactions of myelinating SC with the basal limina.

A library with potential to produce six amino acids cyclic peptides was prepared using pET-28a as the starting plasmid. pVmut was used to amplify the Int(C)-dnaB-N-Int(N) fragment that was inserted into pET28a to give pEV. On pEV, DnaB split intein was expressed under the strong T7 promoter. Analyses of Escherichia coli transformed with pEV showed that DnaB split intein was produced in large quantity and the fusion protein self-spliced efficiently to produce cyclized DnaB-N. A synthesized 115 bp fragment mixture encoding 5 random amino acids was inserted into pEV to generate pEV-IS. The ligation mixture was transformed into E. coli. A library of 10(3) clones was obtained, 20 randomly picked clones were sequenced. All of them contain different sequences. Nine clones were chosen for further analysis. Split-intein-ISs were expressed in large quantity, and 90% of them self-spliced under 16 degrees C in 20 hours. After induction at 30 degrees C for 3 hours, the expressed DnaB split intein was purified using His-column, and then a molecular weight of target cyclic peptide was detected by MALDI-TOF-MS.
Amino Acids ; chemistry ; Base Sequence ; DnaB Helicases ; chemistry ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Inteins ; genetics ; Molecular Sequence Data ; Peptide Library ; Peptides, Cyclic ; chemistry ; Protein Splicing ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Synechocystis ; chemistry