Objective To explore the possible anti-inflammatory mechanism of peroxisome proliferators-activated receptor（PPAR） gamma-agonists by investigating the effects of Rosiglitazone on the expression of phosphorylation of signal transducer and activator of transcription 6（p-STAT6） and the secretion of interleukin（IL）-4 in T-lymphocytes from patients with acute asthma.Methods Peripheral blood T-lymphocytes from 10 healthy volunteers（group A） and 10 patients with acute asthma were isolated，purificated and cultured.T-lymphocytes from the asthma patients were divided into a control group（group B） and a Rosiglitazone treated group（group C）.Rosiglitazone was added with a single dose of 10-4 mol/L at 0 hour of cultrue.After cultured for 48 hours，the concentration of IL-4 in supernatant of each groups were detected by ELISA.The express of p-STAT6 in the T-lymphocytes were determined by Western blot and immunohistochemical techniques.Results The levels of IL-4 were increased markedly in group B than those in group A and group C[（170.34±9.05）pg/mL vs（76.82±7.06）pg/mL and（123.59±8.70）pg/mL，both Plt;0.01]，and which in group C was significantly lower than group A（Plt;0.01）.The levels of p-STAT6 in T lymphocytes were increased markedly in group B than in group A and C[Western blot：（6.28±0.19 vs 3.07±0.18 and 4.12±0.16；immunohistochemistry：（36.58%±7.41）% vs（11.39±4.02）% and（23.92±5.8）%，all Plt;0.01），and which in group C were significantly higher than that in group B（both Plt;0.01）.There was a positive correlation between the level of p-STAT6 and IL-4（Plt;0.01）.Conclusion The levels of p-STAT6 and IL-4 in T-lymphocytes of patients with acute asthma were suppressed by Rosiglitazone in vitro.
Objective To observe the protein expression of c-Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) in normal skin and keloid and to explore their influences on the formation of kloid. Methods Keloid tissues and normal skin tissues were collected from 16 keloid resection patients (experimental group) and 10 voluntary plastic surgery patients (control group). In the experimental group, the keloid formation time ranged from 8 months to 10 years; the keloid tissues were collected from the chest in 6 cases, the ear lobe in 4 cases, the perineum in 2 cases, the shoulder in 3 cases, and the abdomen in 1 case; and all keloid tissues were confirmed by pathological examination. In the control group, normal skin tissues were collected from the abdomen in 4 cases, the thighs in 3 cases, the shoulder in 2 cases, and the back in 1 case. Two-step l ine of Envision immunohistochemical staining was performed to observe the expressions of nonphosphorylated and phosphorylated JNK and ERK; Image Pro Plus 4.5 image analysis system was used to measure the integrated absorbance (IA) and to observe the positive staining strength. Results The immunohistochemical staining showed that no obvious expressions of phosphorylated and non-phosphorylated ERK, JNK were observed in the fibroblasts of the control group, and the expressions of phosphorylated JNK and ERK proteins were significantly higher in the experimental group than in the control group (P lt; 0.05). There was no significant difference in the expressions of non-phosphorylated JNK and ERK proteins between 2 groups (P gt; 0.05). Conclusion Activation of ERK and JNK pathways might be involved in formation of keloid.
Objective To study the function of platelet-derived growth factor (PDGF) in inducing phosphorylation extracellular signalregulated kinase 1/2 (pERK1/2) localization in osteoblasts. Methods Primary osteoblasts were isolated and cultured from cranial bone of 10 mice atthe age of 3 days, weighting 6-9 g without limitation in male and female.The sixth passage osteoblasts were incubated in 1% serum for 12 hours and divided into 2 groups: treated with DMSO(control group) or with PP2(experimentalgroup) for 30 minutes. Each group was further divided into 2 subgroups according to with or without PDGF (20ng/ml) stimulation for 10 minutes. pERK1/2 localization was analysized by immunofluorescence staining in osteoblasts pretreated with or without Src inhibitor PP2. The sixth passage osteoblasts were divided into 2 groups treated with DMSO (control group) or with PP2 (experimental group) for 30 minutes. Each group was further divided into two subgroupsaccording to with or without PDGF (20 ng/ml) stimulation for 10 mintues. The ability of osteoblast migration was determined by wound healing assay. The sixth passage osteoblasts were divided into 2 groups treated with DMSO (control group) or 10 μmol/L PP2 (experimental group) for 30 mintues. Each group was further divided into 2 subgroups according to with or without PDGF (20 ng/ml) stimulation.The pERK1/2 was determined by Western blot in osteoblastic cytoskeleton inducedby PDGF. Results Immunofluorescence staining showed pERK1/2 localization in osteoblastic nuclears and focal adhesions after PDGF stimulation. PP2 significantly inhibited ERK1/2 localization in focal adhesions, but not in nuclears. The wound healing assay results showed that PP2 significantly inhibited osteoblast migration induced by PDGF. The result of Western blot demonstrated that pERK1/2 in osteoblastic cytoskeleton was significantly inhibitedSrc activation is required for pERK1/2 translocalization to focal adhesions and osteoblasts migration.
ObjectiveTo study the expression of urokinase-type plasminogenactivator (uPA) and phosphorylation of glycogen synthase kinase-3β (P-GSK3β) in human colorectal adenocarcinoma and its significance. MethodsSeventy-eight samples of colorectal adenocarcinoma got during operation between January 2006 and December 2010 in Handan Central Hospital were chosen as the study subjects. The immunohistochemical SP method was used to detect uPA and P-GSK3β levels in the 78 cases of colorectal adenocarcinoma, 20 cases of normal colorectal mucosa and 30 cases of colorectal adenoma. ResultsThe positive expression rates of uPA and P-GSK3β in colorectal carcinoma were much higher than those in colorectal mucosa, colorectal polyps, and colorectal adenoma (P<0.05). The expressions of uPA and P-GSK3β were closely correlated with the differentiation, TNM and lymph nodes metastasis (P<0.05). ConclusionThe expression of uPA and P-GSK3β is closely related to the colorectal adenocarcinoma occurrence. Both of them are important biological markers in colorectal adenocarcinoma occurrence and development.
ObjectiveTo explore the effects of inhibition of paxillin phosphorylation on ventilation associated lung injury. MethodsSixty healthy male SD rats were randomly divided into four groups, namely a control group, a protective ventilation group, a high tidal volume ventilation group, and an inhibitor group. The rats in the control group received only tracheotomy and breathe naturally. The rats in the protective ventilation group received protective ventilation for 2 hours. The rats in the high tidal volume ventilation group and the inhibitor group received high tidal volume ventilation for 2 hours. The rats in the inhibitor group additionally received intraperitoneal injection of tyrosine protein kinase inhibitor PP2 before ventilation. All rats were sacrificed and the specimens of lung tissue were collected. The pathological changes of lungs were observed under light microscope and estimated by the diffuse alveolar damage (DAD) score system. The activity of myeloperoxidase (MPO) and the lungs wet/dry (W/D) weight ratio were measured. The expression of tumor necrosis factor-α(TNF-α) in BALF was detected by ELISA. Evans blue (EB) method was used to detect the pulmonary vascular permeability. The expression levels of phosphorylated paxillin (p-paxillin) and paxillin in lung tissue were measured by Western blot. Apoptosis in situ was detected by TUNEL. ResultsThere were significant differences in the W/D ratio, the EB extravasation, DAD score, the MPO activity and the TNF-αexpression in BALF between the high tidal volume ventilation group and the inhibitor group (P < 0.05). The apoptosis rate of each group was sorted from high to low as the high tidal volume ventilation group, the inhibitor group, the protective ventilation group, and the control group. The expression level of p-paxillin was the highest in the high tidal volume ventilation group which was significantly different from other groups (all P < 0.05). There was no significant difference in the expression of paxillin in the protective ventilation group, the high tidal volume ventilation group and the inhibitor group (P > 0.05). ConclusionInhibition of paxillin phosphorylation can significantly alleviate mechanical ventilation associated lung injury.