砄bjective: To determine the effects of interleukin 1 receptor antagonist( IL lra) on the production of IL 6 induced by IL 1? in human gingival fibroblasts(HGFs). Methods:HGFs at passage 5 were exposed to various concentrations of IL 1? with or without IL 1r? . IL 6 in the culture medium was measured with a sandwish ELISA assay. Results:IL 6(?g/L) produced by HGFs exposed to IL 1? at the concentrations (?g/L) of 0.1, 1.0, 10 and 100 were 207?40.29, 235?80.78, 370?40.62, 570?68.17 and 737.5?83.47 respectively. While that by HGFs exposed to IL 1? at 10 ?g/L with IL 1r?(?g/L) at 1, 10, 100 and 1 000 were 387.5?49.69, 312.5?26.81, 242.5?25.86 and 217.5?21.65 respectively. Conclusion: IL lra can inhibite the IL 1? induced IL 6 production in HGFs.

Objective To quickly pinpoint and correct errors in the application level in No.1 Military Medical Project. Methods LogMiner, a powerful tool provided by Oracle was used to mine and analyze the redo log files produced by oracle database in No.1 Military Medical Project. The basic concept, configuration and usage of LogMiner were described through an example of the Register Program in No.1 Military Medical Project. Results The errors made by Register Program was quickly pinpointed and then corrected by SQL statements provided by UNDO_VALUE field. Conclusion The method using LogMiner to pinpoint and correct the errors in the application level in No.1 Military Medical Project is simple, accurate and convenient. It should be a basic technique for maintenance personnel of hospital information system.

The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM's proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM's proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.
Absorbable Implants ; Basement Membrane ; Biocompatible Materials ; chemistry ; Epithelium ; Esophagus ; physiology ; Fibroins ; chemistry ; Humans ; Nanostructures ; chemistry ; Polyesters ; chemistry ; Regeneration ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

BACKGROUND:Polyurethane has good mechanical and physical characteristics and is extensively used in
clinical and experimental studies, but its hydrophobicity and histocompatibility are not ideal, which limits its use in tissue engineering as a biomaterial scaffold to some extents.
OBJECTIVE:To observe the hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin and its compatibility with human hypopharyngeal cel s.
METHODS:The changes in hydrophilicity of polyurethane membrane grafted with silk fibroin and glutin were detected by contact angle measurements. Human hypopharyngeal fibroblasts were cultured in vitro on
polyurethane membrane, silk fibroin-polyurethane membrane, glutin-polyurethane membrane and tissue culture plate. Cel compatibility was compared using cytometry and cel morphology obsevation.
RESULTS AND CONCLUSION:Hydrophilicity of silk fibroin-or glutin-polyurethane membranes significantly increased (P<0.01). The hydrophilicity of silk fibroin-polyurethane membrane was higher than that of
glutin-polyurethane membrane (P<0.01). The number of cel s on the tissue culture plate was the most. The number of human hypopharyngeal fibroblasts on the silk fibroin-or glutin-polyurethane membrane was higher
than that on the polyurethane membrane, especial y on the silk fibroin-polyurethane membrane. These suggested that hydrophilicity and cel compatibility of silk fibroin-or glutin-polyurethane membrane were elevated.

Objective To investigate the effects ofWenyujinessential oil on tau protein phosphorylation in mice with Alzheimer Disease (AD); To discuss the relevant mechanism of action.MethodsSixty Kunming mice were divided into six groups randomly: sham-operation group, model group,Wenyujin essential oil low- and high-dose group, donepezil group andShenzhiling group.β-Amyloid protein (Aβ25-35) was infused into the hippocampal of mice. Mice in each group were given relevant medicine for gavage for 15 d. After the last administration, immunohistochemistry was used to detect the expressions of tau protein phosphorylation points (Ser 404 and Thr 231). Western blot was used to detect tau protein phosphorylation points and the expression of PI3K/Akt protein. Results Compared with sham-operation group, tau protein phosphorylation points in the model group increased (P<0.05), but the phosphorylation levels of PI3K and Akt decreased significantly (P<0.05). Compared with the model group, tau protein phosphorylation (Thr231 and Ser404) inWenyujin essential oil high-dose group decreased significantly (P<0.05), and the phosphorylation levels of PI3K and Akt increased(P<0.01). ConclusionWenyujinessential oil can inhibit tau protein phosphorylation in mice. The possible mechanism may be related to the PI3K/Akt signal pathway.

OBJECTIVETo investigate the effects of bufalin in reversing hepatocyte growth factor (HGF)-induced resistance to afatinib in H1975 lung cancer cells, and explore its possible mechanism.

METHODSThe afatinib-resistant H1975 lung cancer cells (H1975AR) were induced by exogenous HGF and transfected with recombinant adenoviral vector Ad-HGF-GFP. The cytostatic effects of bufalin, afatinib and bufalin plus afatinib on H1975AR cells were evaluated by MTT assay. The impact of combined therapy with bufalin and afatinib on invasion of H1975AR cells was determined by transwell migration assay. The concentrations of HGF in the culture supernatants of H1975/Vec and H1975/HGF cells were determined by ELISA. The expression of EGFR, cMET and EMT signal pathway-related proteins in H1975AR cells treated with bufalin, afatinib and bufalin plus afatinib were detected by Western blot.

RESULTSThe results of MTT assay showed that afatinib did not inhibit the growth of H1975 cells, but after 72 h of the combined treatment with bufalin and afatinib and in the presence of HGF, the growth rate of H1975 cells was (38.67 ± 8.76)%, significantly lower than the growth rate of (63.45 ± 12.65)% in the H1975 cells treated with HGF alone (P < 0.05). The results of transwell migration assay showed that in the presence of HGF, afatinib plus bufalin combination therapy markedly decreased the number of invaded H1975 cells through the Matrigel chamber (48.98 ± 11.43), significantly lower than the 118.92 ± 37.29 of afatinib-treated or the 88.84 ± 19.53 of bufalin-treated cells (P < 0.05 for all). The result of ELISA showed that H1975/HGF cells secreted high levels of HGF, and afatinib and bufalin had no effect on the HGF secretion in H1975/HGF cells. The results of Western blot analysis showed that the expression of p-EGFR, p-cMet, p-AKT, p-ERK, vimentin and snail in H1975AR cells treated with bufalin puls afatinb was down-regulated markedly, and the expression of E-cadherin was up-regulated markedly.

CONCLUSIONSCombination of bufalin and afatinib strongly inhibits the growth of H1975AR lung cancer cells and decreases their invasion ability. The possible mechanism of combined treatment with bufalin and afatinib may be related to the blocking of cMet/PI3K/AKT and cMet/MAPK/ERK pathways and inhibiting of epithelial-mesenchymal transition.

Antineoplastic Agents ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Bufanolides ; pharmacology ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coloring Agents ; Drug Resistance, Neoplasm ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; MAP Kinase Signaling System ; Neoplasm Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; Quinazolines ; pharmacology ; Receptor, Epidermal Growth Factor ; Signal Transduction ; Tetrazolium Salts ; Thiazoles

Objective:To develop an ex vivo normothermic mechanical perfusion(NMP)and compare the effect of air-oxygenated NMP versus oxygen-oxygenated NMP on reducing renal injury from donor after cardiac death(DCD).Methods:All kidneys from DCD rats were subjected to 30 min in situ warm ischemia after cardiac attest.And harvested kidneys were stored for 8h under static cold preservation after NMP for 2h.In experimental groups, kidneys were subjected to either air-oxygenated NMP(group A, n=6)or oxygen-oxygenated NMP(group O, n=6). Sham operation(group C, n=6)and DCD kidneys under static cold preservation without NMP(group SCS, n=6)were employed as controls.The evaluation parameters included creatinine(Cr), aspartate amino transferase(AST)and lactate dehydrogenase(LDH)in perfusate, pathological changes by hematoxylin-eosin(HE)staining, histological criteria, expressions of myeloperoxidase and intercellular adhesion molecular-1(ICAM-1)by immunohistochemistry and Western blot, tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)by enzyme-linked immunoadsorbent assay and level of malondialdehyde(MDA)by thiobarbital method and activity of superoxide dismutase(SOD)by WST-8 in renal tissues.Differences between two groups were analyzed by two-tailed unpaired Student's test and differences among more than two groups by one-way ANOVA.Results:Renal arterial oxygen tensions in NMP perfusate were(576.3±68.2)mmHg with oxygen-oxygenation and(137.0±39.1)mmHg with air-oxygenation.There was significant difference( P<0.05). The pathological injury scores in groups SCS, O and A by HE staining were(7.0±0.1), (5.0±0.9)and(2.5±0.5); injury scores and the expressions of renal proximal tubular epithelial cell vacuolar degeneration in groups O and A were lower than those in group SCS( P<0.05)and injury score in group A was lower than group O( P<0.05). In perfusate, the levels of △Cr, △AST and △LDH in groups O and A were(43.9±52.8)μmol/L and(12.6±3.5)μmol/L, (532.3±52.8)U/L and(49.1±50.4)U/L and(9998.0±2014.4)U/L and(1477.0±810.4)U/L.There were significant differences( P<0.05). In perfused kidneys, the MDA level and SOD activity in groups O and A were(0.192±0.018)mmol/g, (0.162±0.023)mmol/g, (0.6±0.3)×10 3 U/g, (1.7±0.4)×10 3 U/g; TNF-α and IL-6 levels in groups O and A were(124.376±19.635)and(89.331±13.123)ng/g, and(4.038±1.026)×10 3 and(1.774±0.518)×10 3 ng/g.After air-oxygenated NMP, lower renal damage indices were characterized by a lower MDA level and a higher SOD activity, the lower levels of TNF-α and IL-6 and the lower expressions of MPO and ICAM-1 than those in oxygen-oxygenated NMP( P<0.05). Conclusions:NMP with air-oxygenation mimics renal perfusion under physiological conditions and decreases oxidative stress and inflammation injury.It may confer a better retrieval in DCD kidney against warm ischemia injury.

Objective: To test the effect of metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) and/or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods: We transfected HCC827 cells with LV-vector or LV-over/MALAT1. Stable transfected cells (HCC827/Vector, HCC827/MALAT1) were selected by adding puromycin. HCC827/MALAT1 cells were further transfected with the shRNA-negative control (NC) or shRNA-human epidermal growth factor receptor 3 (ERBB3) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib on the proliferation of HCC827 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and/or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib treatment were detected by western blot. Results: The MTT assay showed that sensitivity to osimertinib of HCC827/MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC₅₀) of osimertinib >4 000 nmol/L in HCC827/MALAT1 cells. However, knockdown of ERBB3 facilitated the anti-proliferation effect of osimertinib, and the IC₅₀ of osimertinib in shRNA-ERBB3 cells was (17.27±3.21) nmol/L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827/MALAT1 cells induced by 10 nmol/L osimertinib was (8.38±0.92)%, significantly lower than (27.17±5.83)% of knockdown of ERBB3 (P<0.01). Western blotting showed that the expression of p-ERBB3, p-AKT and p-extracellular regulated protein kinases (ERK) in HCC827/MALAT1 cells was markedly up-regulated, while the expression of p-epithelial growth factor receptor (EGFR) was inhibited. The expressions of p-ERBB3, p-AKT and p-ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p-EGFR, p-ERBB3, p-AKT and p-ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3/PI3K/AKT and ERBB3/MAPK/ERK signaling pathways.