OBJECTIVETo investigate the pro-apoptotic effect of Her-2 targeted recombinant caspase-6 fusion protein on osteosarcoma SOSP-9607 cells.

METHODSRecombinant immunocasp-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody (e23sFv), a PEA translocation domain (PEA aa253-364) and an active caspase-6. The immunocasp-6 gene was cloned into pCMV plasmid to construct a kind of eukaryotic expression vector, i.e. pCMV-e23sfv-PE II-caspase-6 (abbr. pCMV-6) and transfected into SOSP-9607 cells. Murine xenograft models were randomly divided into two groups that received i.m. injections of liposome encapsulated pCMV-6 or pCMV alone. The tumor volume and weight of the nude mice and the tumor weight of the cured mice were observed and statistically analyzed. The morphological changes of the tumors were examined with HE staining, apoptotic morphology of the tumor was observed by TUNEL staining and the gene expression was analyzed by immunohistochemical staining.

RESULTSThe tumor growth of the mice in the treatment group was significantly slower than that of the control group (P = 0.001). The weight of the nude mice in the treatment group was significantly higher than that of the control group (P = 0.0002). The tumor weight of the mice in the treatment group was significantly lower than that of the control group (P = 0.0006). HE and TUNEL staining of the tumor of nude mice in the treatment groups showed typical characteristics of apoptosis, while normal structure was found in the control group. Furthermore, caspase-6 was not found in the tumor and muscle tissues in the control group, but only in the treatment group by immunohistochemistry.

CONCLUSIONImmunocasp-6 can selectively recognize and bind to and kill HER-2 positive osteosarcoma cells, therefore, to offer some foundation for the clinical treatment of osteosarcoma.

ADP Ribose Transferases ; genetics ; Animals ; Apoptosis ; Bacterial Toxins ; genetics ; Bone Neoplasms ; metabolism ; pathology ; Caspase 6 ; genetics ; metabolism ; Cell Line, Tumor ; Exotoxins ; genetics ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Random Allocation ; Receptor, ErbB-2 ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden ; Virulence Factors ; genetics

Currently, hip replacement is an effective treatment for some hip joint diseases. The friction and wear of a prothesis in the human body are the main causes for the failure of joint arthroplasty. It is, therefore, very important to simulate the working conditions of a hip prosthesis in order to get an optimal design and successful clinical applications. This article summarizes wear testing methods of hip prostheses and the developing status of their simulators. Three key aspects of the simulators, i.e., the structures, motions and lubrications, are analyzed in detail. At the end, the developing trend of the simulators are discussed.
Biomechanical Phenomena ; Equipment Failure Analysis ; instrumentation ; Friction ; Hip Prosthesis ; Lubrication ; Materials Testing ; instrumentation ; Prosthesis Design

Objective To study the effects of vascular endothelial growth factor(VEGF)on production of pulmonary surfactants.Method Fetal rat lungs were obtained at 19-day gestation.Primary culture of typeⅡalveolar epithelial cells(AECⅡ)was performed using IgG panning technique.The rats was divided into groups: VEGF,Dexamethasone,VEGF plus Dexamethasone and a control.Total phospholipids,phosphatidylcholine (PC),phosphatidyl glycerol(PG)and sphingornyelin(SM)were determined.Results expressed as mean?SEM. Comparison between groups were made with LSD-t test and one -way ANOVA.Result VEGF,Dexamethasone and VEGF plus Dexamethasone groups showed increased amount of total phospholipids and its compositions on the first day of culture.Conclusions VEGF-165 promotes the production and secretion of pulmonary surfactant. VEGF and Dexamethason may go through different mechanism for enhancement of synthesis of pulmonary surfactant,thereby improve biological function of AECⅡ.

Objective To study the influence of vascular endothelial growth factor(VEGF) on development of alveolar epithelial cell Ⅱ (AECⅡ) and expression of pulmonary surfactant protein B(SP-B) in premature rats.Methods Wistar rats at 19 days gestation were paunched to get embryo and primary AECⅡculture.The rats were divided into 4 groups ,VEGF-165 group,Dexamethasone group,VEGF and Dexamethason group,control group. AECⅡ and SP-B expression were measured by immunology histochemistry.Results SP-B had positive expression in VEGF group, Dexamethason group, VEGF and Dexamethason group. SP-B had negative expression in control group.Conclusion VEGF-165 can increase SP-B positive expression and secret of AECⅡ.VEGF promotes lung maturity.

The present study was undertaken to observe the effect of exogenously administered melatonin on the intensity of β-endorphin (β-Ep) immunoreactivity of the neuron in the arcuate nucleus of rat hypothalamus with an aim to explore the possible mechanisms of the analgesic effect of melatonin. The experimental rats were divided into two groups, one injected intraperitoneally with melatonin (110 mg/kg) and the other with only a vehicle. One hour after injection, the brain was processed for coronal sections, which were stained with immunohistochemical ABC technique. The integral optical density (IOD) and mean optical density (OD) of the stained sections were measured with a computer-assisted image-processing and analytical system. β-Ep immunoreactivity was much decreased in the sections treated with melatonin and the IOD and OD were also decreased significantly (P<0.01; P<0.05). The above results suggest that melatonin may result in a decrease of β-Ep content in the arcuate nucleus, as a result of increased β-Ep release induced by administration of melatonin. It is likely that the analgesic effect of melatonin may be in part mediated by the release of β-endorphin from the arcuate nucleus.

OBJECTIVEThe prevalence of non-alcoholic fatty liver disease (NAFLD) has markedly increased. Insulin resistance has been implicated in the pathogenesis of NAFLD. This study was aimed at observing the relationship between insulin resistance and NAFLD, and evaluating the role of pioglitazone (PGZ) acting as insulin-sensitizing agents in the prevention and treatment of rat fatty liver induced by high fat feeding.

METHODSThe rats were separated randomly into 6 groups: model group I were fed high fat diet for 8 weeks, PGZ prevention group were given PGZ 4 mg/(kg.d) simultaneously, while control group I were fed normal food for 8 weeks; model group II were fed high fat diet for 16 weeks, PGZ treatment group were given PGZ 4 mg/(kg.d) orally simultaneous with high fat diet for 8 weeks after high fat feeding for 8 weeks, control group II were fed normal food for 16 weeks. The rats were sacrificed after 8 weeks and 16 weeks respectively. Liver weight, body weight, serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), tumor necrosis factor alpha (TNF-alpha), fasting blood glucose (FBG), fasting plasma insulin (FINS), HOMA (homeostasis model assessment) insulin resistance index (HOMA-IR), and the liver histology of rats of all groups were assayed.

RESULTSAfter 8 weeks, the liver in model group I showed typical steatosis, accompanied with mild to moderate lobular inflammatory cell infiltration, liver indexes and serum levels of ALT, AST, ALP, TNF-alpha were significantly increased (P<0.05) compared with control group I. Whereas, the degree of hepatic injury was attenuated in PGZ prevention group, liver indexes and serum levels of ALT, ALP were significantly decreased (P<0.05) compared with model group I. After 16 weeks, notable steatosis, and lobular inflammation were observed in model group II rat liver, while the degree of hepatic injury was attenuated in the PGZ treatment group. Liver index, serum levels of ALT, AST, ALP, FINS and HOMA-IR were significantly increased (P<0.05) in model group II compared with control group II. Whereas, in PGZ treatment group, serum levels of AST and FINS showed decreasing tendency, liver indexes, serum levels of ALT, ALP, TNF-alpha and HOMA-IR were significantly decreased compared with model group II.

CONCLUSIONInsulin resistance plays a role in the pathogenesis of NAFLD in rats. Pioglitazone can attenuate insulin resistance and biochemical and histological injury in high fat-induced fatty liver in rats.

Alanine Transaminase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Fatty Liver ; drug therapy ; etiology ; metabolism ; pathology ; Insulin Resistance ; Liver ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Thiazolidinediones ; therapeutic use ; Tumor Necrosis Factor-alpha ; analysis

Animals ; Dietary Fats ; adverse effects ; Fatty Liver ; pathology ; prevention & control ; Hypoglycemic Agents ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Thiazolidinediones ; therapeutic use

OBJECTIVEIn order to explore the role of toll-like receptors 2 (TLR2) in initiating inflammatory response, the expression of TLR2 of the liver and IL-18, TNF-alpha and IFN-gamma of plasma in fulminant hepatic failure was analysed.

METHODSD-galactosamine (D-Gal, 900 mg/kg) and lipopolysaccharide (LPS, 10 microg/kg) were administered intraperitoneally into the BALB/C mice. To evaluate the hepatic injury, serum transaminase (ALT and AST) and plasma IL-18, TNF-alpha and IFN-gamma were determined and the mortality was observed at various time points following the intraperitoneal injection. The level of TLR2 mRNA was measured by semiquantitative RT-PCR. The protein expression of TLR2 in the liver was detected by immunohistochemistry. The data was analyzed by SAS software.

RESULTSAfter 4 hours of intraperitoneal injection of D-Gal/LPS, the serum transaminase and plasma IL-18, TNF-alpha and IFN-gamma levels were elevated. The treated mice began to die at 7 hours. The mortality reached up to 80% at 10 h. TLR2 mRNA was expressed at a low level in liver tissues of normal mice, while it was significantly increased and maintained at a higher level following intraperitoneal injection with D-Gal/LPS. The expression of TLR2 protein was similar to that of the TLR2 mRNA, and the expression of TLR2 mRNA was positively correlated with the concentration of plasma IL-18, TNF-alpha and IFN-gamma (r=0.36, P=0.02; r = 0.48, P 0.003; r = 0.72, P<0.001) at different time points.

CONCLUSIONSOur results showed that TLR2 was involved in initiating and inducing the expression of proinflammation cytokines in this model of fulminant hepatic failure. The results suggest that adjusting the expression of TLR2 might be a new strategy in preventing the development of infectious diseases

Animals ; Galactosamine ; Interferon-gamma ; blood ; Interleukin-18 ; blood ; Lipopolysaccharides ; Liver ; metabolism ; Liver Failure, Acute ; chemically induced ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; biosynthesis ; genetics ; Toll-Like Receptor 2 ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUNDLumiracoxib is a highly selective cyclooxygenase-2 (COX-2) inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms.

METHODSThe antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Drug-drug interactions were analyzed using the coefficient of drug interaction (CDI) to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry.

RESULTSLumiracoxib (15 - 240 micromol/L) has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the IC50 values of 2597 micromol/L and 833 micromol/L, respectively. The synergistic effect was prominent when lumiracoxib (15 - 240 micromol/L) was combined with docetaxol (0.2 - 2 micromol/L) (CDI < 1). Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/G1-phase cells and a decrease of S-phase cells.

CONCLUSIONSLumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.

Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Diclofenac ; analogs & derivatives ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Taxoids ; pharmacology

In order to get some novel compounds with potent iNOS inhibitory activity, 12 target compounds of N-[ 4-( benzimidazole-2-thio) phenyl ] -N'-alkyl guanidine derivatives ( I1- I12 ) were synthesized from 1-benzoyl-3-[ 4-( benzimidazole-2-thio) phenyl] thioureas (4) by hydrolysis with 2. 0 mol x L(-1) sodium hydroxide solution containing tetrahydrofuran to form the corresponding N-[ 4-(benzimidazole-2-thio) phenyl] thioureas (5) which was S-ethylated with ethyl iodide, followed by amination with primary amines or secondary amines. The intermediate 4 was synthesized from 2-mercaptobenzimidazole (1) by reaction with 1-chloro-4-nitrobenzene to form 2-( 4-nitrophenylthio) benzimidazole (2) which was reduced by iron powder and hydrochloric acid, followed by reaction with benzoyl isothiocyanate. The structures of compounds I1 - I12 were confirmed by IR, MS,1H NMR and elemental analysis. The results of preliminary pharmacological test showed that the activities of three compounds (I 1, I8 and I10) were stronger than aminoguanidine, especially for compound I1.
Animals ; Benzimidazoles ; chemical synthesis ; chemistry ; pharmacology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Guanidines ; chemical synthesis ; chemistry ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; enzymology ; Mice ; Molecular Structure ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; metabolism