This study is to explore the interventional effects of fluvastatin on anti-beta2GPI/beta2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-beta2GPI/beta2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR (RT-qPCR) or TF activity was detected by kit. TNF-alpha mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-kappaB p65 and inhibitory protein of NF-kappaB (IkappaB-alpha) were measured by Western blotting. The results suggested that the expression of TF and TNF-alpha on THP-1 cells was significantly up-regulated with treatment of anti-beta2GPI/beta2GPI complex (100 mg x L(-1)), compared with that of untreated cells (P < 0.05). Fluvastatin (50 mg x L(-1)) could decrease TF (mRNA and activity) expression and the level of TNF-alpha (mRNA and protein) in THP-1 cells with anti-beta2GPI/beta2GPI complex. The expression of TF and TNF-alpha was shown in a concentration-dependent manner. Moreover, anti-beta2GPI/beta2GPI complex could downregulate IkappaB-alpha levels and increase the levels of phospho-NF-kappaB p65. And these effects of anti-beta2GPI/beta2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-kappaB signal transduction pathway, thereby inhibit the effects of anti-beta2GPI/beta2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-alpha.

Objective To investigate the application effect of homemade multifunctional prone bed in position nursing for patients with retinal detachment who undergoing vitrectomy with intraocular tamponade .Methods One hundred and sixty-five patients who needed to take the prone position after vitrectomy with silicone oil or gas tamponade were selected and randomly divided into 3 groups:55 patients in the control group1 with the traditional U-shaped soft pillow,55 patients in the control group 2 with the hole bed and U-shaped pillow,55 patients in the experimental group with the homemade multifunctional prone bed .The daily average duration of position, the daily average sleep time,the incidence of head-bit offset, the occurrence of adverse reactions , and the recurrence of retinal detachment were observed and compared among the three groups .Results In the experimental group, the daily average time of maintaining prone position and sleeping was (19.23 ±2.36)h and (8.42 ±0.32)h, respectively, both of which were longer than those in the two control groups , the differences were statistically significant (F=69.806,75.332,respectively;P<0.01).The occurrence of adverse reactions in the experimental group were less than that in the two control groups .The recurrence rate of retinal detachment within 3 months after operation in the experimental group was 1.82%, and lower than that in the two control groups, the difference was statistically significant (χ2 =10.573,P<0.05).Conclusions The application of homemade multifunctional prone bed in patients undergoing vitrectomy with intraocular tamponade can significantly improve the patients'adherence to the prone position, effectively reduce the incidence of postoperative adverse reactions and complications , and more accurately adjust the head position , which can help to promote retinal reattachment and improve success rate of operate .

Aim To investigate SIRT1 activity and an-ti-inflammatory effects of Chikusetsu oleanane saponin ( COS) on the RAW264. 7 macrophage cells induced by lipopolysaccharide ( LPS ) . Methods The nitric oxide ( NO) release was detected with Griess method. Western blot was used to detect the expression of tumor necrosis factor-α( TNF-α) , interleukin 1β( IL-1β) . Nuclear factor-κB ( NF-κB) and silent information ad-justment factor 1 ( SIRT1 ) were tested by immunofluo-rescence. Results 1 mg · L-1 LPS co-cultured with COS at 25~300 mg·L-1 had no significant effect on the growth of RAW264. 7 cells. Compared with the LPS group, COS effectively inhibited the NO release and suppressed the expression of TNF-α and IL-1β, and also inhibited the translocation of NF-κB and up-regulation of SIRT1 . Conclusion COS has protective effects on RAW264. 7 cells stimulated by LPS, which may be related to up-regulating the expression of SIRT1 and promoting the deacetylation of NF-κB, thereby in-hibiting the translocation of NF-κB and reducing the expression of TNF-α and IL-1β.

Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.
Cell Line, Tumor ; Chromatography, Liquid ; methods ; Gene Products, pol ; chemistry ; genetics ; metabolism ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; chemistry ; enzymology ; genetics ; Humans ; Immunoprecipitation ; methods ; Protein Interaction Maps ; Software ; Tandem Mass Spectrometry ; methods

A strain, Pseudomonas sp. X-2-45, with high and stable lipolytical activity was screened by continuously subculturing a lipase-producing bacterium P. sp. LP-1 in culture medium containing Jatropha oil as a sole carbon source. Its hydrolytic activity was 29.79 U/mL, which was increased by 288% as compared to that of parent strain. Furthermore, the growth and lipase synthesis of X-2-45, its catalytic ability to hydrolyze vegetable oils, as well as ester synthesis between fatty acids and organic alcohols were studied. Results showed that rates of bacterial growth and lipase synthesis were significantly raised. Bacterial biomass and lipase activity reached the highest level after 30 h of incubation. Moreover, growth stationary period was prolonged and lipase produced exhibited good stability in culture media during incubation period. Hydro- lytic activity of P. sp. X-2-45 lipase toward Jatropha oil was increased by 378% as compared to parent strain, suggesting that acclimation to Jatropha oil was an effective approach for improving substrate selectivity oflipase. Finally, results of ester synthesis catalyzed by P. sp. X-2-45 lipase indicated that this lipase could catalyze esterification reactions between lauric acid and n-butanol, n-octanol, 1-dodecanol or glycerol, palmitic acid or stearic acid and methanol, n-octanol, 1-dodecanol or glycerol, oleic acid and methanol, n-butanol, n-octanol, 1-dodecanol or glycerol.

Objective To study the adjuvant immunoactivity of polysaccharides from Panax japonicus by alcohol of different concentrations;To discuss its part with the strongest adjuvant immunoactivity. Methods Polysaccharides from Panax japonicus was sunk with alcohol of different concentrations, and 30%alcohol compound, 60%and 90%alcohol polysaccharide were obtained. Different segments of polysaccharide and OVA protein were injected to mice once a week for three times for immunity. Five days after the last immunity, the mice were executed to collect blood, and the antibody titer was determined. The three parts of alcohol compound were scanned by infrared spectrum to determine the type of polysaccharide preliminarily. Results Compared with the control group, the antibody titer of different segments of polysaccharide obviously increased, especially the polysaccharide sunk by 60%alcohol. Infrared spectrum analysis showed that polysaccharides from Panax japonicus contained pyranose ring structure. Conclusion Polysaccharides from Panax japonicus has significant adjuvant immunoactivity, and polysaccharide sunk by 60%alcohol has the strongest adjuvant effects.

OBJECTIVE: To investigate the expression of Myosin VI and Disabled-2 (Dab2) in the cochlea of mice at different ages. METHOD: Forty KM mice were divided into four groups according to age, named as postnatal 2 week (P2w), P5w, P9w, P16month. The localization of protein in the basilar membrane of mice cochlea was detected by immunofluorescence staining and laser scanning confocal microscope (LSCM). The mRNA expression level of protein in cochlear at different ages was evaluated by real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Statistical analysis was performed by the SPSS18.0 software. RESULT: Myosin VI and Disabled-2 protein mainly expressed at the apical cytoplasm of hair cells. As for the inner hair cell, Dab2 labeling was abundant especially at the cuticular plate and nearby. Comparing four immunofluorescence staining images of Myosin VI, we found the fluorescence intensity of P2w and P16m were weaker than that of P5w and P9w. After setting P9w as the control group, qRT-PCR revealed that the mRNA expression of MyosinVI and Dab2 in P2w was less than that in the control group (P < 0.01), while no significant difference was found between P5w and the control group, nor between P16m and the control group (P > 0.05). CONCLUSION Myosin VI and Dab2, two proteins which regulated the clathrin-mediated endocytosis, expressed at hair cells of mice cochlea. In the inner hair cell, this process of endocytosis may be more efficient at the cuticular plate and nearby. The expression level of protein may change in different ages, and this probably leads to a difference of CME, it also may cause a defect of inner hair cells function.
Adaptor Proteins, Vesicular Transport ; metabolism ; Aging ; Animals ; Cochlea ; metabolism ; Endocytosis ; Hair Cells, Auditory ; metabolism ; Hair Cells, Auditory, Inner ; metabolism ; Mice ; Microscopy, Confocal ; Myosin Heavy Chains ; metabolism

Objective To research the protective effects of totalPanax Japonicus extract on learning memory, antioxidation, and anti-apoptosis of aging mice,and explore the mechanism. Methods Male mice were randomly divided into normal group, model group, Vitamin E (VE) group, Panax Japonicus extract low and high dose group. Except for the normal group, the other groups were injected withD-gal on the back of the neck subcutaneously to establish aging model. Normal group and model group were given a gavage with saline and each treatment group was given a gavage with totalPanax Japonicus extract and VE once a day for 7 weeks after the aging model established. All mice were be measured their learning and memory ability in the eighth week. After the test, the morphological changes of CA1 neurons were observed by HE stain. SOD, GSH-Px, MDA levels in brain tissue were measured by biochemical method, the expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR.Results Mice inPanax Japonicus extract low and high dose group could spend less time in searching for the platform, improve the learning and memory ability. TotalPanax Japonicus extract increased the activity of SOD and GSH-Px, while decreased the content of MDA. In addition, it could increase the expression level of Bcl-2 mRNA and reduce the expression level of Bax mRNA as well.Conclusion TotalPanax Japonicus extract has anti-aging effect.

Objective To investigate the expression of adaptin-2(AP-2) in different stages of mouse coch‐lea and its probable role in the auditory generation and formation .Methods Mice were divided into 4 experimental groups by age (7 days old ,15 days old ,35 days old ,16 months old) ,which respectively represented the newborn mice ,developmental mice ,mature mice and old mice .Auditory brainstem response (ABR) ,laser scanning confocal microscopy (LSCM) ,immune-fluroscence histochemistry and qRT - PCR were used in this study .Results For the 15 days old ,35 days old ,16 months old groups ,ABR average threshold was 18 .67 ± 1 .21 dB nHL ,13 .83 ± 1 .47 dB nHL ,37 .83 ± 7 .68 dB nHL ,respectively ,for the 7 days old groups no responses were observed .The AP-2 im‐munoreactivity was found in all the stages of mice cochlea inner hair cell (IHC) cytoplasm ,especially in IHC basal part ,nearby the ribbon synapse .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,immune-flu‐roscence histochemistry IMV(intensity mean value)were 190 .91 ± 17 .27 ,494 .06 ± 27 .63 ,838 .41 ± 38 .23 ,682 .65 ± 72 .22 ,respectively .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,mRNA RQ(relative quantity)were 0 .53 ± 0 .09 ,1 .03 ± 0 .02 ,1 .00 ± 0 .09 ,1 .03 ± 0 .06 ,respectively .Developmental mice expressed significantly higher than those of the newborn in the AP -2 protein expression level which was measured by immuno -fluores‐cence histochemistry and qRT PCR(P<0 .01) .There was no significant difference between old and mature mice in the AP-2 protein expression level measured by qRT PCR (P> 0 .05) ,but mature mice had significant advantages by immuno-fluorescence histochemistry .Conclusion AP-2 protein expression level may closely related to the au‐ditory formation and maintenance because its expression gradually increases with age in mice .

AIM:To study whether inhibition of forkhead box protein M1(FoxM1) sensitizes leukemia K562 cells to homoharringtonine ( HHT ) .METHODS: K562 cells were incubated with HHT at different concentrations ( 0μmol/L, 0.015 μmol/L, 0.030μmol/L and 0.045μmol/L) for different time (0 h, 24 h, 48 h and 72 h).The mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blot.FoxM1 siRNA was transfected into K562 cells with 0.015μmol/L HHT after 6 h.After 72 h incubation, the cell proliferation was detected by cell counting and soft agar assay, and the proportion of apoptotic K562 cells was determined by flow cytometry.The expression of c-Myc and Sp1 were detected by real-time PCR and Western blot.RESULTS:FoxM1 expression was reduced time-dependently and dose-dependently, suggesting that HHT mediated the downregulation of FoxM1 in K562 cells.In K562 cells, treatment with FoxM1 siRNA and HHT inhibited the cell proliferation and promoted the apoptosis significantly.Therefore, inhibition of FoxM1 sensitized leukemia K562 cells to HHT.The expression of c-Myc and Sp1 was positively regulated by FoxM1. CONCLUSION:HHT inhibits Forkhead box protein M1 expression in K562 cells.Inhibition of FoxM1 sensitizes K562 cells to HHT.