Inflammation is a defensive reaction and the most common pathological manifestation of dry eye.In addition,excessive inflammatory response is considered to be the most common pathogenic factor and main cause of dry eye.Currently,the active mechanism of anti-inflammatory drugs has been well-known,and topical antiinflammatory therapy for dry eye is exerting a role at certain extend.However,some adverse responses of these drugs are emerging during the treating procedure.Therefore,it is emphasized that a large sample size of and multicenter randomized-controlled clinical trial is needed to identify the different effects of various anti-inflammatory drugs for different types of dry eye diseases,which will offer a basis for standardized anti-inflammatory treatment for dry eye.

AIM: To explore the skills and characteristics of corneal neovascular model in rat induced by micropocket assay. ·METHODS: Nine eyes of nine Sprague-Dawley rats were studied. Pellets made of vascular endothelial growth factor (VEGF), poly-2-hydroxylethylmethacrylate and sucralfate were implanted into the corneal stroma nocloser than 1mm from the limbus. Biomicroscopic features of corneal neovascular were observed on 1,3, 5, 7th day after the implantation. ·RESULTS: On day 1 after operation, the limbal vessels were dilated, with no angiogenesis appeared. On day 3, angiogenesis began to invade peri-cornea with a brush shape, the area of CNV was (2.23±0.59) mm2. On day 5, new vessels reached the lower margin of pellet densely, the area of CNVwas (6.81±1.35)mm2. On day 7, new vessels continued to elongate, parts of them extended as loops toward the pellet, the area of CNV was (8.92± 1.79)mm2. Neither hyphema or other complications occurred.·CONCLUSION: Corneal neovascular induced by micropocket assay in rat grows steadily, with no complication, and is suitable for quantitative researches.

Background Microvessels are composed of two interacting cell types: endothelial cells and pericytes. Over the past decades, studies of corneal angiogenesis have concentrated mainly on endothelial cells, while interest in pericytes has lagged behind. Objective The present study aimed to investigate the recruitment of vascular endothelial cells and pericytes in rat corneas after alkali burn. Methods Corneal alkali burn models were established in the right eyes of 36 adult SPF SD rats by putting 4 mm medicators containing a 1% 1 mol/L NaOH solution at the central corneas for 30 seconds, and 3 matched normal rats were used as controls. Corneas were excised 1,2,3,5,7 and 10 days after surgery. Frozen sections that parallel with the corneoscleral limbus were constructed. Double immunofluorescence staining was used to observe the dynamic expression of CD31 and α-smooth muscle actin (α-SMA) in corneal tissue for the evaluation of the number of endothelial cells and pericytes. The pericyte coverage index (PCI) was calculated to quantify the recruitment of pericytes to neovascular sites. The use of experimental animals followed the Statement of Association for Research in Vision and Ophthalmology. Results CD31 was expressed in the superficial stromal layer of the cornea on the 1 st day, showing the presence of red fluoresence. The positive cell number for CD31 was gradually increased with the passage of time and proceeded into the deep stromal layer from days 2 through 5 but decreased after that. However,α-SMA was positively expressed on the 2nd day in the cornea after alkali burn with the presence of green fluorescence. The positive cell number for α-SMA was less than those of CD31 throughout the experimental period. The PCI was 0, 16.07%, 11.95%, 43.84%, 73.97% and 86. 21% , respectively, 1,2,3,5,7 and 10 days after surgery. Conclusion Pericytes recruitment to corneal new vessels may play a key role in the stabilization and maturation of angiogeneis.

By Silica gel, Sephadex LH-20 and other materials for isolation and purification and by physicochemical methods and spectral analysis for structural identification, 32 compounds were isolated and identified from ethyl acetate portion of alcohol extract of the Osmanthus fragrans. Their structures were identified as boschniakinic acid (1), ursolaldehyde (2), augustic acid (3), arjunolic acid (4), 5-hydroxymethyl-2-furancarboxaldehyde (5), isoscutellarein (6), 6, 7-dihydroxycoumarin (7), 2α-hydroxy-oleanolic acid (8), quercetin-3-0-β-D-glu-copyranoside (9), D-allito (10), 5, 4'-dihydroxy-7- methoxyflavone-3-0-β-D-glucopyranoside (11), 5,7-dihydroxychromone (12), lupeol (13), naringenin (14), acetyloleanolic acid (15), chlorogenic acid (16), kaempferol-3-0-β- D-glucopyranoside (17), oleanolic acid (18), kaempferol-3-0-β-D-galactopyanoside (19), 3', 7-dihydroxy-4'-methoxyisoflavon (20), ergosta-4,6,8 (14), 22-tetraen-3-one (21), p-hydroxycinnamic acid (22), syringaresinol (23), 3,4-dihydroxyacetophenonel (24), β-sitosterol (25), ethyl p-hydroxyphenylacetate (26), benzoic acid (27), caffeic acid (28), coelonin (29), p-hydorxy-phenylacetic acid (30), p-hydroxyacetophenone (31), and methyl-p-hydroxphenylacetate (32). Except for compounds 2, 4, 5, 8-11, 13, 15, 18, 20, 25, and 27, the rest were isolated from the Osmanthus fragrans for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Oleaceae ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To investigate the influence of iodine excess on expression of TRAIl/TRAIL-sR1 in NOD and Balb/c mice and to study the effect of TRAIl/TRAIL-sR1 on the pathogenesis of experimental autoimmune thyroiditis(EAT). Methods Both Balb/c and NOD mice were divided randomly into control and iodine excess group by feeding with water containing no NaI or 0.05% Nal. The mice were sacrificed after 8 weeks. TRAIL and TRAIL-sR1 mRNA levels were detected by RT-PCR. The function, morphology and apoptosis of thyroids were also observed by ELISA and Tunnel stain. Results Treated by HI, enlarged follicles and flattened epithelium by accumulation of colloid were found in thyroids of both NOD and Balb/c mice. But significant lymphoid cell infiltration and local fibrosis were only found in thyroids of NOD HI group. The relative weight of thyroids of NOD mice in HI group[(104.8±14.5)mg/kg]was heavier than that of control group [(71.8±20.4)mg/kg]. The level of TT4 declined in HI group[(30.77±3.59)mmol/L]compared with control group[(36.43±2.66)mmol/L], meanwhile, the level of TSH was higher in HI group[(6.98±0.66)μg/L]than that in control group [(5.55±0.56)μg/L]. The difference being statistically significant(t=7.773,-9.526,-4.458, all P < 0.05). The relative weight of thyroids of Balb/c mice of HI group[(155.8±20.8)mg/kg]also heavier than that of control group [(105.1±22.0) mg/kg]. The level of TT4 droped in HI group [(19.75±3.32) mmoL/L]was higher than that in control group[(23.46±6.21)mmoL/L], the level of TSH in HI group[(4.14±1.71)μg/L]was higher than that in control group[(3.55±1.41)μg/L], the difference being statistically significant(t=7.554,-7.239,3.140, all P< 0.05). A great deal of apoptotie ceils observed in NOD (3.97±0.91) and Balb/c mice (1.05±0.45) by Tunnel stain were greater than control groups (0.21±0.15, 0.10±0.03), the difference being statistically significant in beth of the two species(t=-7.167,-17.772, both P < 0.05). The apoptosis index of thyroid follicular epithelium in NOD was obviously higher than Balb/c(t=-7.625, P<0.05). The level of TRAIL mRNA did not remarkably change in Balb/c between control group(0.000 59±0.000 39) and HI group(0.001 24±0.000 46, t=-1.940, P>0.05), but it increased apparently in NOD mice HI group(0.018 88±0.005 77) than that of control group(0.009 61± 0.00591, t=-2.71, P<0.05). The level of the expression of TRAIL-sR1 mRNA increased in HI groups of NOD (0.000 53±0.000 15) and Balb/c mice(0.000 42±0.000 09) than that in control groups of NOD(0.000 28± 0.000 05) and Balb/c mice (0.000 17±0.000 06) and the differences were statistically significant between the two species(t=3.050,3.990, all P<0.05). The differences of the expression of TRAIL and TRAIL-sR1 mRNA between the two species were significant(t=-3.37,-4.76, all P<0.05). Conclusions Iodine excess induces colloid goiter in beth species of mice and thyroiditis in NOD mice. The increase of TRAIL and TRAIL-sR1 influenced by iodine excess is one of the molecular bases of follicular epithelium apoptosis and inflammation in thyroids. Genetic factor is a key factor in the pathogenesis of thyroiditis.

BACKGROUNDMenin is a ubiquitously expressed protein encoded by the multiple endocrine neoplasia type 1 (MEN1) gene. Besides its importance in endocrine organs, menin has been shown to interact with the mixed lineage leukemia (MLL) protein, a histone H3 lysine 4 methyltransferase, and plays a critical role in hematopoiesis and leukemogenesis. Previous studies have shown that menin promotes transforming growth factor beta (TGF-β) signaling in endocrine cells. However, little is known regarding the impact of TGF-β pathway on menin in hematopoietic system. Here, with leukemia cell lines generated from conditional MEN1 or TGF-β receptor (TβRII) knockout mouse models, we investigated the possible cross-talk of these two pathways in leukemia cells.

METHODSMEN1 or TβRII conditional knockout mice were bred and the bone marrow cells were transduced with retroviruses expressing oncogeneic MLL-AF9 (a mixed lineage leukemia fusion protein) to generate two leukemia cell lines. Cell proliferation assays were performed to investigate the effect of TGF-β treatment on MLL-AF9 transformed leukemia cells with/without MEN1 or TβRII excision. Menin protein was detected with Western blotting and mRNA levels of cell proliferation-related genes Cyclin A(2) and Cyclin E(2) were examined with real-time RT-PCR for each treated sample. In vivo effect of TGF-β signal on menin expression was also investigated in mouse liver tissue after TβRII excision.

RESULTSTGF-β not only inhibited the proliferation of wild type MLL-AF9 transformed mouse bone marrow cells, but also up-regulated menin expression in these cells. Moreover, TGF-β failed to further inhibit the proliferation of Men1-null cells as compared to Men1-expressing control cells. Furthermore, excision of TβRII, a vital component in TGF-β signaling pathway, down-regulated menin expression in MLL-AF9 transformed mouse bone marrow cells. In vivo data also confirmed that menin expression was decreased in liver samples of conditional TβRII knockout mice after TβRII excision.

CONCLUSIONThese results provided the first piece of evidence of cross-talk between menin and TGF-β signaling pathways in regulating proliferation of leukemia cells, suggesting that manipulating the cross-talk of the two pathways may lead to a novel therapy for leukemia.

Animals ; Blotting, Western ; Cells, Cultured ; Humans ; Leukemia ; metabolism ; Mice ; Mice, Knockout ; Multiple Endocrine Neoplasia Type 1 ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Transforming Growth Factor beta ; genetics ; metabolism ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVE: To investigate the expression of c-fos in rats' skin during wound healing. METHODS: Immunohistochemistry was conducted on paraffin section from incised wounding model of rat skin. RESULTS: Fos protein improved from the time of 10 min after wounding in the wound edge, then it reached peak at 3 h. 24 h after injury, the quantity of Fos expression had no difference with that of normal skin. CONCLUSION Fos is sensitive after wound, but should be used with other criteria in wounding interval estimation as it's unstediness.
Animals ; Genes, Immediate-Early ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-fos/biosynthesis* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Time Factors ; Wounds and Injuries/metabolism*

BACKGROUNDCorneal neovascular leakage can lead to edema and secondary scarring. Previous studies have shown that pericytes play a key role in maturation of angiogenesis. The present studies investigate the relationship between vascular permeability and pericyte coverage of endothelial cells in rat corneal neovascular induced by alkali burns.

METHODSCorneal neovascular vessels induced by alkali burns was performed in Sprague-Dawley rats. Corneas were excised on 1, 2, 3, 5, 7 and 10 days after cauterization. The vascular permeability rate was measured by the Evans blue method. The microvessel pericyte coverage index (MPI) was applied to quantify the pericyte coverage through double immunofluorescent staining of frozen sections of corneas with CD31 as the endothelial and alpha-smooth muscle actin (alpha-SMA) as the pericyte markers. The correlation between permeability rate and MPI was analyzed. Pericyte coverage was confirmed ultrastructually using transmission electron microscopy.

RESULTSThe vascular permeability rate was (1.14 +/- 0.17), (0.24 +/- 0.08), (0.29 +/- 0.16), (0.14 +/- 0.10), (0.09 +/- 0.06) and (0.05 +/- 0.04) microg x ml(-1) x mm(-2) respectively on 1, 2, 3, 5, 7 and 10 days after cauterization. The MPI was 0, 16.07%, 11.95%, 43.84%, 73.97% and 86.21% respectively at the above mentioned time points. The correlation coefficient between MPI and the permeability rate was -0.943 (P = 0.005).

CONCLUSIONSPericyte recruitment was significantly correlated with the permeability of corneal neovascularization induced by alkali burns in rats. Therapeutic strategies aiming at anti-leakage should be most effective if they promote pericytes proliferation in the course of corneal neovascularization.

Alkalies ; Animals ; Burns, Chemical ; physiopathology ; Capillary Permeability ; Cell Movement ; Cornea ; blood supply ; ultrastructure ; Corneal Neovascularization ; physiopathology ; Eye Burns ; chemically induced ; physiopathology ; Female ; Fluorescent Antibody Technique ; Pericytes ; physiology ; Rats ; Rats, Sprague-Dawley

Adult ; Chylous Ascites ; therapy ; Female ; Fibrin Tissue Adhesive ; therapeutic use ; Humans ; Middle Aged ; Parenteral Nutrition, Total ; Somatostatin ; therapeutic use

OBJECTIVETo study the influence of Kai1/CD82 transfection on the growth, adherence, separation and invasion potential of LoVo colon carcinoma cell line.

METHODSKai1/CD82 cDNA was transfected into LoVo cells, and a stable expressing clone was established. In vitro methodology was used to obtain the growth curve and also to detect the adherence, separation and invasion potential of the transfected LoVo cells, in comparison with those of control cells without transfection.

RESULTSCompared with the control, no change was observed in the growth pattern of transfected LoVo cells. The numbers of adherent cells in the two groups were 0.08, 0.63, 0.83, 0.91 (x 10(5)) for the transfected cells and 0.04, 0.48, 0.71, 0.82 (x 10(5)) for the control cells respectively after 10, 20, 30, 40 minutes culture with shaking. The difference at 20, 30 and 40 minutes was statistically significant (P < 0.05). The separation rates of each group were 13%, 20%, 53% for the transfected cells and 11%, 28%, 60% for the control cells, respectively after 5, 10, 15 minutes culture with shaking. The difference at 10 and 15 minutes was statistically significant (P < 0.05). The aggregation rate of the transfected cells was higher than that of the control cells after culture with mild shaking for 5 hours (64.8% vs. 58.6%, P < 0.05). After co-incubation with endothelium cells ECV304, the number of invading cells decreased more in the transfected cells than that in the control cells (6.33/field and 17.67/field, P < 0.05).

CONCLUSIONTransfection expression of Kail/CD82 into LoVo cell line results in an increase of cell adherence and aggregation, but a diminished capability of separation and invasion, suggesting that the expression of Kai1/CD82 gene may inhibit the metastatic potential of colon carcinoma.

Antigens, CD ; Cell Adhesion ; Cell Division ; Colonic Neoplasms ; genetics ; pathology ; prevention & control ; Genes, Tumor Suppressor ; Humans ; Kangai-1 Protein ; Membrane Glycoproteins ; genetics ; Neoplasm Invasiveness ; Proto-Oncogene Proteins ; Transfection