Objective:To investigate the expression of u-PA and PAI-1 and their possible role in oral submucous fibrosis(OSF). Methods:Immunohistochemistry and imaging analysis were used to detect the expression of u-PA and PAI-1 in 30 cases of OSF and 10 cases of normal control.Results:The expression of u-PA and PAI-1 was observed in the cytoplasm of blood vessel endothelium,fibroblasts and inflammatory infiltration cells in lamina propria of OSF and control samples.u-PA expression was increased at the early stage of OSF(P0.05).PAI-1 expression was increased following the development of the disease(P

Objective To investigate the changes in the serum MMP-9 (matrix metalloproteinase-9) and the expressions of MMP-9 in lung, kidney and intestine in rats with multiple organ dysfunction syndrome (MODS) and confirm extracellular matrix injuries being the mechanism in MODS in order to propose a novel theoretical basis for cfinical treatment of MODS. Method Forty wister rats were randomly divided into two groups: control group (n=8) and MODS model group (n=32). The rats of model group were further divided into four subgroups ac-cordingto the time elapsed after modelling: 12 h (n=8), 24 h(n=8) ,48 h(n=8) and 72 h (n=8), and were modelled by celiac injection of mixed liquid of zymosan-paraffin (4 mL/100 g) after blood loss (1mL/100 g) by extirpating their left eyes. Blood,lung, kidney and intestine were sampled 12,24,48 and 72 hours after models were established. The histological changes in the lung, kidney and intestine of the rats were observed by light mi-croscope. The serum MMP-9 were measured by enzyme-linked immunosorbent assay (ELISA). The immunohisto-chemistry was used to observe the expression of MMP-9 in lung,kidney and intestine during different phases of MODS. The data were processed by one-way ANOVA and Bivariate analysis. Results Compared with control group, the organs were injured by congestion, edema and inflammatory cells infiltration to a certain extent in model groups. The serum MMP-9 increased markedly 12 hours after modelling (P<0.01 ) and peaked 48 hours later. The expressions of MMP-9 in lung, kidney and small intestine significantly increased from 12 h to 72 h after mod-elling (P<0.01 or 0.05). Conclusions The MMP-9 increased both in serum and tissue are closely associated with the pathological process of MODS. The mechanism of organ damage probably attributes to the damage of extra-celluar matrix and tissue construction.

Graphene ( GN) and multiwalled carbon nanotubes ( MWCNT) composites were coated on glassy carbon electrode ( GCE ) and then poly ( nicotinic acid ) ( PNA ) was electrodeposited on the modified electrode. The electrochemical behavior of pyridoxine hydrochloride ( VB6 ) was investigated at the modified electrode by cyclic voltammetry ( CV ) and differential pulse voltammetry ( DPV ) . Results showed the oxidation current of VB6 at the GN-MWCNT/PNA/GCE was obviously larger than that at GCE, PNA/GCE and GN/MWCNT/GCE. The oxidation process of VB6 was an irreversible diffusion-controlled process involving one electron and two protons. The liner range between the peak current intensity of DPV and the concentration of VB6 was 0 . 05-200 μmol/L with a detection limit of 0 . 02 μmol/L ( S/N=3 ) . The modified electrode showed a good reproducibility with a relative standard deviation of 3 . 1% ( n=8 ) . The proposed method was applied to the analysis of vitamin B6 in vitamin B6 tablets and compound vitamin B tablets with recoveries between 96 . 1%-104 . 5%.

Objective To investigate the expression and the effects of tissue inhibitor of metalloproteinases-1 (TIMP-1) on lungs of rats with sepsis.Methods Forty Sprague-Dawley (SD) rats were randomly divided into two groups,namely sham group (n =8) and sepsis model group (n =32).The rats of model group were modeled by cecal ligation and puncture (CLP),and were further divided into four subgroups as per the time after modeling,namely 6 h (n =8),12 h (n =8),24 h (n =8),48 h (n =8)subgroups.Blood and lung samples were taken 6 h,12 h,24 h and 48 h after modeling.The histological changes in lungs of the rats were observed under light microscope.Expressions of TIMP-1 mRNA,Bax mRNA and Bcl-2 mRNA in lungs were measured by RT-PCR.The immunohistochemistry was used to label the CD18 in lungs during different phases of sepsis.The data were processed by t test.Results Compared with sham group,the lung tissues of rats in model group were injured to a certain extent after CLP.The expression of TIMP-1 mRNA and the number of CD18 positive cells increased at the same time (P ＜ 0.01),and peaked 24 hours later (P ＜ 0.01).While the expression of Bax mRNA in model group decreased markedly 12-48 hours after modeling (P ＜ 0.01-0.05),and reached minimum 48 hours later (P ＜ 0.01).The expression of Bcl-2 mRNA in model group changed unnoticeable.The positive correlation between variations in number of CD18 positive cells and expression of TIMP-1 mRNA was found in model group (r =0.426,P ＜ 0.01).Conclusions The increase in expression of TIMP-1 mRNA in lungs is closely associated with the lung injury of sepsis.The mechanism of lung injury is likely attributed to the preservation of inflammatory cells from apoptosis,and the persistent inflammation response causes tissue damage,leading to organ dysfunction.

Objective To explore the effects of peroxisome proliferator-activated receptorγ (PPARγ) agonist rosiglitazone and 15-deoxy-delta-12,14-prostaglandin J2 (15d-PGJ2) on the expression of PPARγ, toll-like receptor 4 (TLR4) and the activation of STAT1 as well as the local inflammation reaction of abdominal cavity in sprague dawley (SD) rats with peritoneal dialysis- related acute peritonitis induced by lipopolysaccharide (LPS). Methods Twenty-four male SD rats were equally randomized to four groups(n=6 each): control group, injected with 4.25% dextrose peritoneal dialysate (PDF) via abdominal cavity(90 ml/kg); LPS group, injected with LPS(1 mg/kg) via abdominal cavity 4 hours later follewed by PDF injection; rosiglitazone plus LPS group (Rosi group), preconditioned with rosiglitazone (20 mg·kg-1·d-1) by intragastric way for 3 days, then injected with LPS and PDF via abdominal cavity; 15d-PGJ2 plus LPS group (15d-PGJ2 group), preconditioned with 15d-PGJ2 (0.3 mg·kg-1·d-1)via abdominal cavity injection for 3 days, then injected with LPS and PDF via abdominal cavity. The rats were killed 4 hours after PDF injection, IL-6 level in abdominal dropsy was determined by ELISA. Peritoneum tissue was stained by Masson. Leucocyte count in abdominal dropsy was performed. The mRNA expression of PPARγ and TLR4 in peritoneum tissue was determined by RT-PCR; the protein expression of PPARγ, TLR4, p-STAT1 and STAT1 in peritoneum tissue was analyzed by Western blot. Results IL-6 level of abdominal dropsy in LPS group [median 268.53 (range 201.87-335.19) ng/L] was significantly higher than that of control group [median 147.62 (range 130.60-164.64) ng/L] (P<0.01). The IL-6 level of abdominal dropsy in Rosi group [median 110.20 (range 77.60-142.80) ng/L] was significantly lower than that of LPS group (P<0.05). Compared to that of control group, the edematous degree of peritoneum in LPS group was significantly severer, meanwhile, mRNA and proteins expression of PPARγ and TLR4 in rat peritoneum were also significantly higher (P<0.05, P<0.01). Compared to that of LPS group, the edematous degree of peritoneum in Rosi group was lighter, the expression of PPARγ and TLR4 mRNA was significantly up-regulated (P<0.05), meanwhile their proteins expression was down-regulated (P<0.05); and in 15d-PGJ2 group, the edematous degree of peritoneum, the expression of PPARγ mRNA and protein was also decreased (P<0.05), but TLR4 mRNA expression was up-regulated (P<0.01), however, its protein expression was down-regulated (P<0.05). There were no significant differences in leucocyte count of abdominal dropsy among the four groups. The p-STAT1 expression in the rats peritoneum induced by LPS was markedly increased by both rosiglitazone and 15d-PGJ2 (P<0.01). Conclusions Both rosiglitazone and 15d-PGJ2 can down-regnlate the inflammatory reaction in rat peritonitis induced by LIPS, which may be involved in modulating the expression of associated functional protein during LPS signal pathway.

ated acute peritonitis induced by LPS in rats.

AIM:To study the effects and mechanism of recombinant human defensin ?1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS:The influences of defensin ?1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase(MEK)inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin ?1,the phosphorylation of extracellular signal regulated kinase(ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS:Defensin ?1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h(P20 mg/L decreased cell proliferation.The cell proliferation induced by defensin ?1 was inhibited by U0126.Stimulation of the cells with defensin ?1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin ?1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h(P

To investigate the effects of leptin on expression of acyl-coenzymeA: cholesterol acyl-transferases-1 (ACAT-1) in monocyte-macrophage differentiation, human monocytic cells (THP-1) were cultured in RPMI 1640 and made to differentiate into macrophages under the incubation with phorbol myristate acetate (PMA) for 48 h. The cells were divided into 4 groups according to different intervention factors as follows: MCs cultured in RPM11640 medium with 10% FBS for 48 h served as MC group (control group), MCs cultured in medium with serum-free RPM11640 containing 5% BSA, 100 nmol/L PMA for 48 h as MP group, MCs cultured in RPMI1640 medium with 10% FBS, 10 micromol/ml leptin for 48 h as leptin-MC group, and MCs cultured in medium with serum-free RPMI1640 containing 5% BSA. 100 nmol/L PMA, and 10 micromol/ml leptin for 48 h as leptin-MP group. Immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR) and Western blot were performed, respectively, to observe the effects of leptin on expression of ACAT-1 in the monocyte-macrophage differentiation. Our results showed that expression of ACAT-1 protein and mRNA in MP-group is two times that in MC-group (P<0.05), and the expression of ACAT-1 protein and mRNA increased by up to 4 folds in leptin-MP group-as compared with that of MC group (P<0.01). Thus, our results support the idea that expression of ACAT-1 increases more in cultured human macrophages than in monocytes, and leptin can significantly promote ACAT-1 expression. It was concluded that high expression of ACAT-1 may accelerate the development of human atherogenesis, and leptin might participate in atherogenesis by increasing expression of ACAT-1.
Atherosclerosis ; enzymology ; Cell Differentiation ; Cells, Cultured ; Humans ; Leptin ; pharmacology ; Macrophages ; cytology ; enzymology ; Monocytes ; cytology ; enzymology ; Sterol O-Acyltransferase ; biosynthesis ; genetics ; Tetradecanoylphorbol Acetate

Objective To investigate the effects of microRNA-205-3p (miR-205-3p) and microRNA-205-5p (miR-205-5p) on the cell proliferation,migration and invasion of head and neck squamous cell carcinoma (HNSCC).Methods The miRNA expression data were obtained from The Cancer Genome Atlas,and were used to determine miR-205 expression in HNSCC and paracancerous tissues.Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-205-3p and miR-205-5p in HNSCC cell lines 5-8F,6-10B,CNE2,Tu686 and mucosal tissues of nasopharynx.MiR-205-3p inhibitor,miR-205-5p inhibitor and miR-NC were transfected into Tu686 cells,and qRT-PCR was employed to evaluate the silence efficiency.In the following study,the cells were divided into four groups:miR-205-3p inhibitor and control,miR-205-5p inhibitor and control.Then the cell counting kit (CCK-8) assay,Transwell migration and invasion assays were carried out to examine the proliferation,migration and invasion abilities of the cells in the four groups.Results Compared with paracancerous tissues,the higher expression of miR-205 in HNSCC tissues was observed [M (QR):61 012 (51 448) vs.28 579 (35 959),Z =-6.420,P ＜ 0.001)].The expression levels of miR-205-3p in HNSCC cell lines 5-8F,6-10B,CNE2,Tu686 and mucosal tissues of nasopharynx were 0.36 ± 0.07,0.20 ± 0.06,0.15 ± 0.04,0.25 ± 0.04 and 1.00 ±0.00 respectively,with a significant difference (F =162.71,P ＜ 0.001).The expression levels of miR-205-5p in cell lines 5-8F,6-10B,CNE2,Tu686 and mucosal tissues of nasopharynx were 0.20 ±0.01,0.21 ±0.01,1.06 ±0.18,23.61 ±2.07 and 1.00 ±0.00 respectively,with a significant difference (F =371.81,P ＜0.001).Compared with 5-8F,6-10B,CNE2 cells,the expression of miR-205-3p in Tu686 cells was higher than those in 6-10B,CNE2 cells (P =0.195;P =0.020),and lower than that in 5-8F cells (P =0.023),and the expression of miR-205-5p in Tu686 cells was the highest (P ＜ 0.001;P ＜ 0.001;P ＜ 0.001).The expressions of miR-205-3p and miR-205-5p in Tu686 cells were effectively downregulated by inhibitors,and the silence efficiencies were 87％ and 83％,respectively.The absorbance (A) values of miR-205-3p inhibitor group on the first,second,third,fourth and fifth day were 0.26 ± 0.06,0.55 ± 0.11,1.52 ±0.13,1.91 ± 0.07,2.14 ± 0.24,and those of miR-NC group were 0.29 ± 0.07,0.78 ± 0.11,1.59 ±0.15,1.95 ±0.08,2.02 ±0.12.There were no significant differences between the two groups (t =0.506,P=0.639;t =2.459,P=0.070;t =0.573,P=0.597;t =0.655,P=0.548;t=-0.759,P=0.490).The cell proliferations of miR-205-5p inhibitor group on the first,second,third,fourth and fifth day were 0.30 ± 0.08,0.61 ± 0.08,0.85 ± 0.08,1.08 ± 0.12,1.16 ± 0.18,and those of miR-NC group were 0.41 ± 0.10,0.78 ± 0.14,1.33 ± 0.28,1.87 ± 0.09,2.08 ± 0.19.The cell proliferations of miR-205-5p inhibitor group on the third,fourth and fifth day were significantly lower than those of miR-NC group (t =3.665,P =0.017;t =12.223,P ＜ 0.001;t =7.825,P ＜ 0.001).Transwell migration assay demonstrated that downregnlation of miR-205-3p had no significant effect on migration abilities of Tu686 cells.The numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 192.00 ± 28.49 and 188.40 ± 22.52,respectively.There was no significant difference between the two groups (t =-0.160,P =0.877).Downregulation of miR-205-5p significantly decreased the migration abilities of Tu686 cells.The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 109.40 ± 27.63 and 183.60 ± 31.63,respectively,and the difference was statistically significant (t =3.951,P =0.004).Transwell invasion assay showed that the numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 93.40 ± 10.24 and 96.20 ± 16.56,respectively.There was no significant difference between the two groups (t =0.322,P =0.756).The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 53.00 ± 17.80 and 94.40 ± 14.38,respectively,and the difference was statistically significant (t =4.045,P =0.004).Conclusion Downregulation of miR-205-5p can inhibit the proliferation,migration and invasion abilities of Tu686 cells.However,downregulation of miR-205-3p has no significant effect on the proliferation,migration and invasion of Tu686 cells.

OBJECTIVE: To investigate the expression and biological significance of HMGB1 and VEGF protein in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and further study the correlation between HMGB1 and VEGF protein. METHOD: The expression of HMGB1 and VEGF protein was evaluated by immunohistochemical staining in 69 cases of LSCC specimens and 15 cases of adjacent epithelial tissue samples, and futher correlated with clinicopathologic parameters. RESULT: The positive rates of HMGB1 and VEGF in LSCC tissues were significantly higher than those in adjacent non-cancerous mucosa (P < 0.01), and the expression of these two marks was closely correlated with clinical stage (P < 0.05) and metastasis (P < 0.05) in LSCC. While the expression of HMGB1 and VEGF had no significant correlations with age, sex, histological differentiation and tumor site (P > 0. 05). There was a positive correlation between the expression of HMGB1 and VEGF (P < 0.05). The Kaplan-Meier survival analysis showed that patients with strong expression of HMGB1 or VEGF had poorer overall survival compared with that in patients with relative low HMGB1 or VEGF expression (P < 0.05). Multivariate COX regression analysis revealed that both lymph node metastasis and HMGB1 expression were independent prognostic factors for patients with LSCC. CONCLUSION This study demonstrated that HMGB1 and VEGF protein overexpression were closely associated with clinical stage, metastasis and poorer prognosis in patients with LSCC. Increased expression of these two proteins in LSCC suggested that HMGB1 and VEGF might play a critical role in the initiation and progression of LSCC.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; HMGB1 Protein ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Vascular Endothelial Growth Factor A ; metabolism