Objective To study the expression of S100A8 and the relationship between S100A8 and Toll-like receptor 4 (TLR4) in focal cerebral ischemia reperfusion (I/R) injury.Methods C3H/HeJ mice with TLR4 gene mutation (n =30) and C3H/HeN with normal TLR4 gene mice (n =30) were divided into 4 groups at random (random number),namely C3H/HeJ model group (n =18),C3H/HeJ control group (n =12),and C3H/HeN model group (n =18).C3H/HeN control group (n =12).Middle cerebral artery was occluded to make I/R model in mice by using thread embolism method.Brain tissues were collected after ischemia for one hour and reperfusion for 12 hours.Stroke outcome was evaluated by determination of infarct volume of brain tissue and assessment of neurological scores.And brain injury after cerebral I/R was observed by optical microscope after TTC and HE staining.The immunofluorescence technique and RT-PCR were used to determine the protein level and expression of S100A8 mRNA in damaged brain tissues.Results Compared with C3H/HeN model mice,TLR4-deficient model mice (C3H/ He J) had lower infarct volumes and better outcomes of neurological function after resuscitation for 12 hours.Compared with control groups,the expression of S100A8 mRNA and level of S100A8 protein increased greatly in damaged brain tissues of model mice after I/R injury.In addition,model mice with lacked TLR4 (C3H/HeJ) had lower expression of I/R-induced S100A8 mRNA than C3H/HeN mice in model group,indicating that the close relationship between the levels of S100A8 and TLR4.Conclusions S100A8 interaction with TLR4 might be involved in brain damage and in inflammation triggered by I/R injury.

Aim To investigate the mechanism of dia-betes changing the hepatic CYP1 A2 through in vitro cell culture study.Methods The function of CYP1 A2 in HepG2 and Fa2N-4 cells were evaluated by determi-ning the level of phenacetin metabolism,and the mR-NA expression of CYP1 A2 in cells was detected by real time PCR.HepG2 cells were co-cultured with serum of diabetic rats(type 1 and type 2)and normal rats,then the CYP1 A2 function in cells were evaluated.Then, the HepG2 and Fa2N-4 cells were co-cultured with a series of concentrations of saturated (including palmitic acid and stearic acid)and unsaturated fatty acids(in-cluding oleic acid and linoleic acid)for 48 h,and the function and expression of CYP1 A2 in the cells were compared.Results It was found that the activities of CYP1 A2 were higher in cells incubated with diabetic serum of both type.All high concentration of fatty acids could increase the function and expression of CYP1 A2 in both HepG2 and Fa2N-4 cells.Conclusion It is speculated that the abnormal level of fatty acids under diabetic state might be part of the reasons why diabetes change the hepatic CYP1 A2,which provides the basis for future study.

BACKGROUND:Recent studies have showed that S100A8 has been implicated in the pathobiology of inflammatory disorders, and that cerebral ischemia reperfusion (I/R) rapidly activates inflammation responses via Toll-like receptor 4 (TLR4). This study aimed to explore the expression of S100A8 and the relationship between S100A8 and TLR4 in focal cerebral ischemia reperfusion injury.METHODS:C3H/HeJ mice (n=30) and C3H/HeN mice (n=30) were divided randomly into a C3H/HeJ model group (n=18), a C3H/HeJ control group (n=12), a C3H/HeN model group (n=18), and a C3H/HeN control group (n=12). Middle cerebral artery I/R model in mice was produced using a thread embolism method. The brains of the mice were collected after ischemia for 1 hour and reperfusion for 12 hours. Stroke outcome was evaluated by determination of infarct volume and assessment of neurological impairment scores. Brain injury after cerebral I/R was observed by an optical microscope after TTC and HE dyeing. The immunofluorescence technique and real time PCR were used to test the expression level of S100A8 in brain damage.RESULTS:Compared with C3H/HeN mice, TLR4-deficient mice (C3H/HeJ) had lower infarct volumes and better outcomes in neurological tests. The levels of S100A8 increased sharply in the brains of mice after I/R injury. In addition, mice that lacked TLR4 (C3H/HeJ) had lower expression of I/R-induced S100A8 than C3H/HeN mice in the model group, indicating that a close relationship might exist between the levels of S100A8 and TLR4.CONCLUSION:S100A8 interaction with TLR4 might be involved in brain damage and in inflammation triggered by I/R injury.

Objective To assessed the expression of inducible costimulator(ICOS)on peripheral blood and joint fluid CD4,CDS,CD45RO T cells and B cells in rheumatoid arthritis(RA).Methods Expression of ICOS and ICOS/CD45RO on peripheral blood and joint fluid CD4~+CD8~+T cells and ICOS ligand(ICOSL)on CD19 B cells from RA patients and healthy volunteers were determind by three-color flow cytometry.Compar- ision with active and inactive RA,initial and relapsed RA had been done.Results Joint fluid CD4 and CD8 T cells expressing ICOS,ICOS/CD45RO were significantly increased than peripheral blood in RA patients and healthy subjects.Joint fluid B cells expressing ICOSL were significantly reduced than peripheral blood in RA patients.Meanwhile,peripheral blood B cells expressing ICOSL were significantly reduced in active RA than inactive RA patients.Conclusion Hyperexpression of ICOS and ICOS/CD45RO on joint fluid CD4 and CD8 T cells and lowexpression of ICOSL in B cells from RA patients,expecially in active RA may contribute to the local immunopathological roles and joint destructions in the pathogenesis of RA.

Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.

Chronic intermitted hypoxia including sleep breathing disorder leads to brain injury. This study explores the potential therapeutic effects of grape seed proanthocyanidin as a neuroprotective agent. A rat model of chronic intermittent hypoxia was employed, and the animals were given low or high doses of grape seed proanthocyanidin. The ultrastructure changes in the brain, the biochemical components, and the animal behavior were examined. The results showed that with hypoxia exposure, neuronal mitochondria exhibited injuries at ultrastructural level, with increased malondialdehyde (MDA) content and reduced superoxide dismutase (SOD) activity. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining revealed increased cell apoptosis in hippocampus. In Morris water maze the animals showed decreased learning abilities, when compared to normal control. The administration of grape seed proanthocyanidin treatment reversed all these observed changes, and improved the learning behavior. We concluded that grape seed proanthocyanidin could alleviate the brain injury caused by hypoxia from sleep breathing disorder.

OBJECTIVETo analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry.

METHODS2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions.

RESULTS2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated.

CONCLUSIONThe effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.

Cobalt ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Proteome ; analysis ; Proteomics ; Umbilical Cord ; cytology ; drug effects

BACKGROUND: Corneal repair materials can be used as an alternative of human donor corneas to repair corneal injuries, but their evaluation of effectiveness is necessary before entering clinical trials. Unfortunately, there is no standardized method for effectiveness evaluation until now. OBJECTIVE: To establish and validate a corneal fungal infection model in rabbits and the corneal transplantation method. METHODS: Twelve New Zealand White rabbits were selected to establish a corneal fungal infection model in the left eye (experimental) and a normal control in the right eye. Two weeks after modeling, acellular porcine corneal stroma was transplanted into the left eye. After transplantation, slit lamp microscope test, corneal thickness detection, intraocular pressure measurement, confocal microscopy test and optical coherence tomography were performed. Then the degree of transparency, degree of epithelium healing, degree of edema, degree of corneal neovascularization and degree of material thawing were evaluated. The corneal pathological sections with hematoxylin-eosin staining were observed at 3, 6 and 12 months after surgery. RESULTS AND CONCLUSION: (1) The corneal thickness increased significantly at 1 month after transplantation, varied slightly within 3-6 months, and became close to the normal value at 1 year. (2) The intraocular pressure of the left eye was close to normal eyes. (3) Findings from the optical coherence tomography showed that the repair materials fit well with the implantation bed at 7 days after transplantation; the transplanted area was fully covered with epithelial cells at 6 months after transplantation, and the uniform thickness of the repair material in the transplanted area was detected; the grafted cornea was restored to normal cornea at 1 year after transplantation. (4) Under the confocal microscope, the repair materials in the transplanted area were evenly spread at 1 month after transplantation; few cells migrated into the transplanted area at 6 months after transplantation; the density of epithelial cells was increased, and there were migrated cells in the transplanted cells, but the cell number was less than that of normal eyes at 1 year after transplantation. (5) The corneal repair material was almost completely transparent at 1 year after transplantation, indicating its effectiveness in the treatment of infectious corneal ulcers. No rejection occurred, indicating that the corneal healing material is well-curative. (6) At 3 months after transplantation, a large number of stromal cells migrated to the corneal substitute, and the collagen fibers in the transplanted area were arranged neatly and densely without obvious scarring and degradation. At 6 months after transplantation, the transplanted area basically recovered. At 1 year after transplantation, the transplanted area was fully restored to the normal cornea state with good biocompatibility. Our experimental findings indicate that the rabbit model of corneal fungal infection and the corneal transplantation method can be used to evaluate the effectiveness of such corneal materials.

Photosynthetic and growth characteristics of Angelica dahurica were studied in order to clarity the relations of photosynthesis, growth and root dry weight, and provide a theoretical basis for its cultivation. Photosynthesis and growth indexes were meas- ured every 25 days. The contents of chlorophyll a, b, a + b, soluble protein and the activities of Hill reaction, Ca(2+)-ATPase, Mg(2+)-ATPase had an increasing trend; They had the highest value in leaf high-speed growth period. Then, they were decreased in root high- speed growth period. The root dry weight showed negative corelation with photosynthetic characteristics indexes except stomatal con- ductance, however, the negative corelation only from net photosynthetic rate and Ca(2+)-ATPase were significant. The vegetative growth period of spring sowing A. dahuricia was divided into three phases: seedling period, leaf high-speed growth period and root high-speed growth period. The root dry weight showed a significantly positive corelation with the root diameter, leaf dry weight, shoot dry weight, aboveground dry weight. There was the competitive relation between aboveground and underground, so underground growth could be es- timated from leaf area and shoot dimeter.
Adenosine Triphosphatases ; metabolism ; Angelica ; enzymology ; growth & development ; metabolism ; Chlorophyll ; metabolism ; Photosynthesis ; Plant Proteins ; metabolism ; Plant Roots ; growth & development ; Seasons