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Objective: To investigate inhibition of dihydroartemisinin （DHA） on contact hypersensitivity （CHS） in mice and its mechanisms. Methods: Mice were sensitized with 0.5%2, 4-dinitrofluorobenzene （DNFB） on shaved abdominal flank skin for 2 consecutive days, and CHS was elicited by 0.25%DNFB on left ear 5 days later. Right ear was treated with acetone/olive-oil alone as control. Mice were given DHA orally 2 days before sensitization. Skin histopathological changes were observed by HE staining, infiltrations of CD4+T and CD8+T cells in ear skin were observed by immunohistochemical staining, and spleen index was detected. Serum IL-6, IFN-γ, IL-10, TGF-β and monocyte chemotactic protein （MCP）-1 changes were detected by ELISA, skin Smad2 and Smad3 phosphorylation levels were detected by Western blot, and skin and spleen immune cells infiltration were detected by flow cytometry. Results: DHA significantly improved ear swelling, skin erythema and spleen index of CHS mice （P<0.05）. Histopathological analysis revealed that degree of edema and cellular infiltration were markedly decreased in DHA-treated CHS mice. Flow cytometry results show that DHA treatment significantly decreased CD4+T cells, CD8+T cells, dendritic cells and macrophages infiltration in ear skin and spleen （P<0.05）. ELISA results showed that DHA treatment also diminished serum levels of IL-6, IFN-γ, TGF-β and MCP-1 compared with model group （P<0.05）. Western blot showed that Smad2 and Smad3 phosphorylation were normalized by DHA treatment （P<0.05）. Conclusion: DHA suppresses CHS by reducing infiltration of immune cells and regulating TGF-β/Smad signaling pathway, which provides a new drug selection and experimental basis for CHS treatment. [ABSTRACT FROM AUTHOR]

BACKGROUND: Cervical degenerative ossification may aggravate nerve compression in patients with cervical degenerative disease, and also make the decompression procedure more challenging. However, the accompanied histological changes of the intervertebral disc and the underlying mechanism of degenerative ossification have not yet been documented. OBJECTIVE: To explore the histological features of the cervical disc in patient with cervical degenerative ossification and the underlying mechanisms of local ossification. METHODS: Patients with cervical spondylosis undergoing surgical treatment were selected from the clinical practice, and cervical disc samples were harvested during the surgical process. Based on preoperative cervical X-ray and computed tomography (CT) examinations, all the disc samples were divided into ossification group and non-ossification group. Hematoxylin-eosin staining, Masson trichrome staining and Safranin O-fast green staining were used to compare the histological differences between the two groups. Furthermore, for the expression of transforming growth factor-β1, p-Smad2 and p-Smad3 in the disc and osteophyte samples, a semi-quantitative immunohistochemistry method was used to compare the difference between the two groups. RESULTS AND CONCLUSION: All the disc tissues could be clearly divided into the outer annulus fibrosus, the inner annulus fibrosus and the nucleus pulposus. Mature trabeculae and bone marrow cavities were detected in all ossification samples. Histologically, the number of disc cells in the ossification group was significantly higher than that of the non-ossification group (P < 0.05). However, the content of proteoglycan in the matrix was significantly lower than that in the non-ossification group (P < 0.05). Transforming growth factor β1/Smad2/3 was detected in all the disc and osteophyte tissues. Local content of transforming growth factor β1, p-Smad2 and p-Smad3 in the ossification group was significantly higher than that of the non-ossification group (P < 0.05). All the findings indicate that, compared with the non-ossification group, cervical degenerative ossification in patients in the ossification group is more severe. Higher expression of transforming growth factor β1 in the local microenvironment may promote the development of degenerative ossification. However, downstream-activated p-Smad2 and p-Smad3 may play different roles in the development of degenerative ossification. [ABSTRACT FROM AUTHOR]

Objective To investigate the influence of medicated serum of Houshi Hei San (Hou's Black Powder, HHS-MS) on expressions of phospholipase Cγ2 (PLCγ2), extracellular regulated kinase (ERIK), transforming growth factor-- (TGF-β) and Smad2/3 in human umbilical vein endothelial cells (HUVEC) after injury of oxygen-glucose deprivation (OGD). Methods HUVEC were randomly divided into control group, model group, 2.5% HHS-MS group, 5.0% HHS-MS group, 10.0% HHS- MS group and 20. 0% HHS-MS group. The OGD model was established through putting HUVEC in a hypoxic incubator for continuous cultivation for 6 h after changed to sugar-free nutrient solution in model group and all HHS-MS groups. HUVEC were cultured normally in the same duration. After modeling, the nutrient solution was changed to complete nutrient solution with 10% blank serum in control group and model group, and MS with corresponding mass fraction of HHS-MS was used in all HHS-MS groups. The HUVEC total protein was extracted after normal cultivation for 24 h. The protein expressions of PLCγ2, ERK, p-ERK, TGF-β and Smad2/3 in HUVEC were detected by using Western blotting assay. Results Compared with control group, the expression of PLC-γ2 and Smad2/3 were up-regulated (P < 0.01) and expression of TGF-β was down-regulated (P <0.01) in model group. Compared with model group, the expression of TGF-β was up-regulated in 5. 0% HHS-MS group and expressions of PLC-γ2 and Smad2/3 were down-regulated in 2. 5% HHS-MS group, 5. 0% HHS-MS group and 10. 0% HHS-MS group (P < 0. 01 or P < 0. 05). Conclusion HHS-MS can regulate PLCγ2/ERK signaling pathway and TGF-β/Smad2/3 signaling pathway, and improve the proliferation of HUVEC after OGD injury. [ABSTRACT FROM AUTHOR]

Objective To investigate the mechanism of angiotensin II (AngII) in inducing pulmonary fibrosis in rats. Methods Thirty male SPF male Wistar rats were divided into control group, model group, and model+ AngII subgroup, with 10 rats in each group. HE staining was performed in each group to evaluate the degree of pulmonary fibrosis and alveolar inflammation. RT-PCR was used to detect mRNA relative expression levels of Yesrelated protein 1 (Yap1), PDZ-binding transcriptional coactivator (TAZ), Smad2/3 and type I collagen (Col-I). Results In the lung tissue of the control group, the alveolar wall was intact under the light microscope, the structure was clear, the alveolar septum was not widened, there was no inflammatory exudate in the bronchial cavity or alveolar cavity, and the alveoli had no inflammatory changes. In the model group, fusion of some alveoli was observed under the light microscope, the lung tissue structure was disordered, the alveolar septum was severely congested, with edema and inflammatory cell infiltration. Under the light microscope, the lung tissue of the model +AngII group showed uniform thickening of the lung and increased inflammatory cell infiltration. The scores of pulmonary fibrosis and alveolar inflammation in the model group and model+AngII group were significantly higher than those in the control group (P<0.05), and they were significantly higher in the model+AngII group than in the model group (P<0.05). The mRNA and protein of Yap1, TAZ, Smad2/3 and Col-I in the model group and model+AngII group were significantly higher than those in the control group (P <0.05). The expressions were significantly higher in the model+AngII group than in the model group (P <0.05). Conclusion AngII may promote the synthesis of type I collagen by up-regulating the expression of Yap1 in the Hippo signaling pathway and the expression of Smad2/3 in the TGF-β1/Smad signaling pathway, and finally induce pulmonary fibrosis in rats. [ABSTRACT FROM AUTHOR]

Objective ： To investigate the protective effects of Jinlida granules (JLD) on the liver injury in a rat model of type 1 diabetes mellitus (DM) and the possible mechanism involved. Methods ： The SD diabetes model rats were randomly divided into the diabetes model group (or DM group) and the Jinlida granule treatment group (or the JLD group)，each consisting of 5 rats，ancl another 5 rats were assigned as the normal control group (or the NC group). For the rats in the NC，DM and JLD groups， 5% carboxy methyl cellulose (CMC), 5% CMC and JLD suspensions at a dosage of 3g/kg were administered by gavage once a day for 8 weeks. The levels of fasting blood, glucose (BG) and glycosylated hemoglobin (HbAic) were detected at hour 24 following the last medication • The blood levels of hyaluronic acid (HA)，W type collagen (IY-C)，procollagen EI (PC-IE) and laminin (LN) were analyzed by radioimmunoassay. The activities of ALT ， AST, alkaline phosphates (AKP) in blood and the content of angiotensin II (Ang II) in the liver tissue were measured by ELISA • The expression levels of transforming growth factor-ft (TGF-pi)protein and p-Smad2/3 proteins in the liver tissue were detected by Western blot. And the expression levels of Ang II mRNA，Smad2 mRNA and Smad3 mRNA in the liver tissue were detected by real time PCR. Results ： When comparisons were made between the DM and NC groups，the levels of BG， HbAic，ALT，AST，AKP， HA， LN，PC-IE and ff-C were elevated significantly (P

Objective To study the effect of paeonol (PAE) and PNS on the expression of transforming growth factor (TGF)- beta 1/ Smad2/3 pathway in rats with acute myocardial infarction (AMI), and the possible molecular mechanism thereof. Methods Model of AMI was made using left anterior descending coronary branch ligation. According to the intervention methods rats were divided into model group, PAE group (8 mg·kg-1), PNS group (40 mg·kg-1), PAE (4 mg·kg-1) + PNS (20 mg·kg-1) low dose group, PAE (8 mg·kg-1) + PNS (40 mg·kg-1) high dose group and captopril positive control group (10 mg·kg-1). Rats without ligation were used as Sham operation group. Left ventricular systolic blood pressure (LVSP), left ventricular diastolic pressure (LVEDP) and the maximum rise and fall rate (/dtmax DP) were detected after 28-day treatment. HE staining was used to observe changes of myocardial tissue. The protein expression levels of TGF-β1 and Smad2/3 were detected by Western blot assay. Results There were significant differences in parameters used for detecting treatment group and model group, formula group and single drug group, formula high dose group and formula low dose group (P < 0.01). The model group showed pathological changes. All treatment groups showed different degrees of pathological improvement. There was the most significant improvement in formulae group and captopril group. Compared with the model group, TGF-β1 and Smad2/3 protein expressions were decreased in treatment group. The expression levels of TGF-β1 and Smad2/3 were significantly decreased in formula group than those of PAE group and PNS group, and lower levels in formula high dose group than those of formula low dose group (P < 0.05). Conclusion Paeonol and PNS can inhibit the expressions of TGF-β/ Smad 2/3 protein in rats with AMI, by blocking TGF-β / Smad pathway. [ABSTRACT FROM AUTHOR]

PURPOSE: To investigate the effect of different concentrations of fluoride on the expression of Smad2/3 which is a specific intracellular signal transduction molecule of TGF-β, and to explore the mechanism of dental fluorosis in rat. METHODS: Forty Wistar rats were randomly divided into 3 groups. HE and immunohistochemistry were used to detect the changes of the ameloblasts and the expression of Smad2/3 in rat incisors. MetaMorph microscope images analysis system and SPSS 12.0 software package were used to analyze the images and data. RESULTS: Typical symptoms of dental fluorosis were found in the fluoride group. The expression of Smad2/3 in the ameloblasts in the experimental group was significantly lower than that in the control group (P<0.01); but the difference was not significant between the low-dose group and high-dose group (P>0.05). CONCLUSIONS: By inhibiting the expression of Smad2/3 in ameloblasts, fluoride affects the differentiation and development of enamel, leading to the occurrence of dental fluorosis in rat. Supported by National Natural Science Foundation of China(30600509) and Natural Science Foundation of Liaoning Province(20102278). [ABSTRACT FROM AUTHOR]

目的 探讨TGF-β1/Smad2/3通路在亚低温状态保护大鼠急性脑缺血再灌注损伤的作用. 方法 选择健康成年雄性SD大鼠30只,制作急性脑缺血再灌注损伤大鼠模型,随机分为假手术组、手术组、亚低温组各10只. 亚低温组在亚低温处理1 h后采用干湿法测定各组脑组织含水量,Q-PCR和Western blotting分别检测各组脑组织TGF-β1 和p-Smad2/3的基因和蛋白的表达量. 结果 亚低温组脑组织含水量低于假手术组、手术组( P均<0.05),而TGF-β1 和p-Smad2/3的基因和蛋白的表达量高于假手术组、手术组(P均<0.05). 结论 亚低温能减轻急性脑缺血再灌注损伤引起的急性脑水肿,TGF-β1/Smad2/3信号通路参与了此过程. [ABSTRACT FROM AUTHOR]