Objective To investigate the influence of benzopyrene on extracellular matrix(ECM) protein deposition of human airway smooth muscle cells(HASMC) and the related pathway mechanism.Methods HASMC were primarily cultured and the 2-6 generations cells were applied in this experiment.The expression amount of ECM gene and protein was detected by real time PCR and Western Blot the phosphorylation level was analyzed by using the Western Blot method.Results Benzopyrene could to increase the expression of HASMC collagen Ⅰ α1 protein (P＜0.01) and ECM protein (including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA(P＜0.05).Benzopyrene could induce the rapid increase of ERK1/2 phosphorylation level (P＜0.01).Furthermore,the ERK pathway inhibitor PD98059 could significantly inhibit the increase of benzopyrene-induced collagen Ⅰ α1 (P＜0.01)and ECM protein(including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA expression(P＜0.01).Conclusion Benzopyrene induces the ECM protein deposition of HASMC by activating the ERK1/2 pathway,blocking the ERK1/2 signal pathway can inhibit the benzopyrene-induced airway remodeling.

Objective To investigate of the effect and mechanisman of SERCA2 on the phenotype modulation of HASMCs. Methods HASMCs were starved for 5 days and divided into different groups ,then we observed morphology change of the cells from the microscope and detected a-actin、SERCA2 and p-ERK by Western Blot,cells proliferation was observed by CCK-8 method. Results Compared with the control group,PDGF could reduce a-actin of HASMCs and increased the cells proliferation ability ,TSG could significantly inhibit the effect (P<0.01), PDGF could also significantly inhibit SERCA2 protein and increased the expression p-ERK (P<0.01), while U0126 significantly inhibited the effect (P < 0.01). Conclusion PDGF may induce HASMCs phenotype modulation through the regulation of SERCA2 and p-ERK.

Dust ; analysis ; Humans ; Magnoliopsida ; Pneumoconiosis ; epidemiology ; Prevalence

Objective To investigate the effect of statin on trabecular bone microstructure by using trabecular bone score (TBS), a new type of bone microstructure evaluation index. Methods A total of 253 middle and aged patients hospitalized in the First Affiliated Hospital of Nanjing Medical University between January 2014 and March 2016 were retrospectively analyzed. According to whether statin was used or not, patients were divided into two groups: 90 patients in the statin use group (statin was use for more than 1 year) and 163 in the control group (not taken any statin). Serum biochemical indicators, such as triacylglycerol, total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, alkaline phosphatase, fasting blood glucose and 25 hydroxy vitamin D, were compared between the two groups. Dual energy X-ray absorptiometry (DXA) was used to measure the bone mineral density (BMD) of lumbar spine and femoral neck. TBS was calculated with TBS iNsight? software, and the DXA image of lumbar spine were analyzed. Results Values of total cholesterol and low density lipoprotein cholesterol were significantly lower in statin group compared with those of control group (P<0.001), while no significant differences in other biochemical indicators between two groups (P>0.05). There was higher lumbar spine BMD statin group compared to that of control group (g/cm2:1.04 ± 0.19 vs. 0.96 ± 0.14, P<0.01). There was higher lumbar spine BMD and higher TBS (1.31±0.09 vs. 1.26±0.09, P<0.01) in statin group than that of control group. There was no difference in the femoral neck BMD between the two groups (P>0.05). Conclusion Statin increases lumbar spine BMD and improves trabecular bone microstructure in middle and aged people.

Objective: To investigate the expression changes of TNF ? mRNA after brain explosive injury in dogs. Methods: With the brain explosive injury model we had developed, nuclei acid in situ hybridization with an antisense RNA probe and computer image analysis were used to detect the dynamic changes of TNF ? mRNA in the cerebral cortex, hippocampus, hypothalamus, and up brain stem after brain injury. Results: The expression of TNF ? mRNA could be found in several regions of normal dog's brain tissues, and the expression became more obvious and widespread following explosive injury to the brain. In all of the brain regions, the expression changes of cerebral contussive area and hippocampus were more significant. TNF ? mRNA expression increased significantly at 30 min and reached maximum at 1 h after the explosive injury, and then decreased gradually, which was higher than normal's. More expression were detected in the ipsilateral hemisphere than contralateral hemisphere. Conclusion: The expression of TNF ? mRNA in basal condition implicate that TNF ? may play an important role in neural function. It is suggested that TNF ? may play an important role in traumatic brain injury. [

Nanocavity biomaterials with recognition specificity imprinted by using proteins as templates may successful serve as substitutes for antibodies, enzymes, and other native biological structures as well as cell bracket materials. It has numerous applications in biotechnology, medicine and so on. In this paper, the principle of template imprinting is introduced briefly, the specialty of template imprinting of proteins is analyzed, and the methods of template imprinting of proteins including protein entrapment, microbead surface imprinting, flat surface imprinting as well as the epitope are reviewed in details.
Animals ; Biocompatible Materials ; chemical synthesis ; Biotechnology ; Genomic Imprinting ; Humans ; Macromolecular Substances ; Nanotechnology ; Peptides ; chemical synthesis ; Polymers ; Proteins ; chemical synthesis ; Templates, Genetic

Objective:To investigate the expression changes at the transcriptional level in normal lung tissues of mice after exposure to heavy ion radiation for different durations at different doses, aiming to provide evidence for exploring sensitive genes of heavy ion radiation, heavy ion radiation effect and the damage mechanism.Methods:Experiments on the temporal kinetics: the whole thorax of mice was irradiated with 14.5Gy carbon-ions and the total RNA of lung tissue was extracted at 3days, 7days, 3 weeks and 24 weeks. In dose-dependent experiment, the total RNA of lung tissue was extracted at 1 week after irradiated with a growing thoracic dose of 0, 7.5, 10.5, 12.5, 14.5, 17.5 and 20Gy. Protein-to-protein interaction (PPI) analysis and gene-ontology biological process enrichment analysis were performed on significant differentially expressed genes (DEGs).Results:A clearly differential expression patterns were observed at 3-day (acute stage), 1-week (subacute stage), 3-week (inflammatory stage) and 24-week (fibrosis stage) following 14.5Gy carbon-ions irradiation. Among those, the 3-day time point was found to be the mostly different from the other time points, whereas the 7-day time point had the highest uniformity with the other time points. Cellular apoptosis was the main type of cell death in normal lung tissues following carbon-ions exposure. The interactive genes of Phlda3, GDF15, Mgmt and Bax were identified as the radiosensitive genes, and Phlda3 was the center ( R=0.76, P<0.001). Conclusion:The findings in this study provide transcriptional insights into the biological mechanism underlying normal lung tissue toxicity induced by carbon-ions.

OBJECTIVETo investigate the expression and release of high mobility group Box-1 protein (HMGB1) in the lung tissue of mice with respiratory syncytial virus (RSV) infection.

METHODSEighteen mice were randomized into PBS control group, RSV group and RSV/ribavirin group. Seven days after RSV infection in the mice in the latter two groups, the bronchoalveolar lavage fluid (BALF) was collected for cell counting and classification, and the levels of IL-4, IFN-gamma and HMGB1 in the supernatants of the BALF were detected. The left lungs of the mice were harvested for pathological examination with HE staining, and the right lungs were taken for detecting the expression of HMGB1 by Western blotting.

RESULTSRSV induced a TH1 inflammation in the lung tissue as shown by significantly increased IFN-gamma and decreased IL-4 levels in the BALF. The total BALF cells, neutrophils and macrophages in the RSV group were significantly higher than those in the control group (P<0.05), and the cell counts were significantly decreased by ribavirin treatment (P<0.05). HE staining showed neutrophil and lymphocyte infiltration in the lumen and submucous layer of the airway in RSV group. The level of HMGB1 in the BALF significantly increased in the RSV group as compared with that in the control group (P<0.05), but was lowered by ribavirin treatment (P<0.05). The expression of the HMGB1 in the lung tissue significantly increased in the RSV group in comparison with that in the control group (P<0.05), and was not significantly decreased by ribavirin treatment (P>0.05).

CONCLUSIONSThe increased expression and release of HMGB1 in the lung tissue of mice with RSV infection is probably involved in the development of RSV infection-related lung diseases.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; HMGB1 Protein ; biosynthesis ; genetics ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Respiratory Syncytial Virus Infections ; metabolism

OBJECTIVETo evaluate the therapeutic effect of in vitro induced autologous bone marrow-derived liver stem cell transplantation for posthepatitic cirrhosis.

METHODSBetween Jun 2008 and Mar 2009, 12 patients with posthepatitic cirrhosis and portal hypertensive underwent azygousportal disconnection and splenectomy in our department. The patients were then divided into two groups to receive autologous bone marrow-deprived liver stem cell infusion via the hepatic artery after in vitro induction for 7 days (n=6) or saline (n=6). The therapeutic effects of the operations on the liver functions and liver fibrosis index were evaluated.

RESULTSAll the patients recovered uneventfully and no side effect of the operation was found. After the operation, the patients receiving bone marrow-deprived liver stem cell infusion showed better hepatic function improvement than those receiving saline infusion (P<0.05).

CONCLUSIONTransplantation of in vitro induced autologous bone marrow-derived liver stem cell via the hepatic artery is safe and effective for treatment of posthepatitic cirrhosis.

Adult ; Bone Marrow Cells ; cytology ; Female ; Hepatitis, Viral, Human ; complications ; Humans ; Liver Cirrhosis ; etiology ; therapy ; virology ; Male ; Middle Aged ; Stem Cell Transplantation ; Transplantation, Autologous

OBJECTIVETo investigate the effect of toluene diisocyanate (TDI) on the production of reactive oxygen species (ROS) and the permeability of human bronchial epithelial (HBE) cells.

METHODSTDI-human serum albumin (TDI-HSA) conjugate was prepared using a modified Son's method. MTT assay was used to assess HBE cell viability after exposure to different concentrations of TDI-HSA. The level of intracellular ROS of HBE cells was detected by flow cytometry with an oxidation-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) uploading, and the permeability of cell monolayer was assessed by detecting the transepithelial electrical resistance (TEER).

RESULTSThe exposure to 120 µg/ml TDI-HSA did not obviously affect the cell viability. Compared with the control group, the intracellular fluorescent intensity increased significantly in the cells exposed to 20, 60, and 100 µg/ml TDI-HSA (P<0.05). The intracellular ROS production increased significantly after 100 µg/ml TDI-HSA treatment (P<0.05), but the increment in ROS production was significantly suppressed by pretreatment of the cells with N-acetylcysteine (NAC) (P<0.05), which also enhanced the TEER decreased by TDI-HSA treatment (P<0.05).

CONCLUSIONSTDI enhances the permeability of HBE cell monolayer partially through a ROS-mediated pathway, suggesting the importance of oxidative stress in TDI-induced pulmonary diseases.

Bronchi ; cytology ; Cell Line ; Cell Membrane Permeability ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Humans ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Serum Albumin ; pharmacology ; Toluene 2,4-Diisocyanate ; pharmacology