Aim To observe the effects of receptor interacting protein 2(RIP2)on macrophage inflammatory ac-tivation and polarity transformation,and to explore the mechanism of RIP2 in macrophage phagocytosis of oxidized low den-sity lipoprotein(ox-LDL).Methods THP-1 derived macrophages were treated with different doses(10,25 and 50 mg/L)of ox-LDL for 24 hours,and treated with 50 mg/L ox-LDL for 8,16 and 24 hours.Real-time quantitative PCR and Western blot were used to detect the expression of RIP2 mRNA and protein in THP-1 derived macrophages,and ELISA was used to detect the secretion of tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein-1(MCP-1).Three pairs of RIP2 siRNA were designed,transfecting them into cells using hiperfict transfection reagent,real-time quanti-tative PCR and Western blot were used to detect the expression of RIP2 mRNA and protein in THP-1 derived macrophages after transfection,in order to screen for the optimal siRNA transfection concentration and the most effective pair of siRNA.After transfection with the most effective RIP2 siRNA,cells were treated with 50 mg/L ox-LDL for 24 hours,ELISA was used to detect the secretion of TNF-α,MCP-1,interleukin-10(IL-10)and interleukin-12(IL-12),real-time quantitative PCR was used to detect the expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1),flow cytometry was used to detect the expression of cell surface antigens CD86,CD80 and CD163.Results Ox-LDL induced the ex-pression of RIP2 in macrophages in a dose-dependent and time-dependent manner.With the increase of ox-LDL treatment dose and time,the expression of RIP2 mRNA and protein increased.Among them,the expression of RIP2 protein in the 50 mg/L group was 7.6 times of the control group,and the expression of RIP2 protein in the 24 h group was 17.9 times of the control group(P＜0.001).The ELISA results showed that with the increase of ox-LDL treatment dose and time,the secretion of TNF-α and MCP-1 increased(P＜0.05).After transfection of RIP2 siRNA into cells,ELISA results showed that the secretion of TNF-α,MCP-1 and IL-10 in the ox-LDL group was 2.4 times,2.9 times and 1.8 times of the control group(P＜0.01),and the secretion of IL-12 decreased by 34.2％compared to the control group(P＜0.01);the secretion of TNF-α,MCP-1 and IL-10 in the siRNA+ox-LDL group decreased by 37.4％,45.3％and 27.4％,respectively,com-pared to the ox-LDL group(P＜0.01),while the secretion of IL-12 increased by 31.4％(P＜0.05).The results of flow cytometry and real-time quantitative PCR showed that the expression of CD86,CD80 and iNOS mRNA in the ox-LDL group was 14.2 times,33.8 times and 4.5 times of those of the control group,respectively,while the expression of CD163 and Arg-1 mRNA decreased by 33.4％and 43.0％,respectively,compared with the control group(P＜0.05);the expression of CD86,CD80 and iNOS mRNA in the siRNA+ox-LDL group decreased by 27.6％,29.3％and 34.3％,respectively,compared to the ox-LDL group,while the expression of CD163 and Arg-1 mRNA increased by 30.3％and 38.6％,respec-tively(P＜0.05).Conclusion RIP2 expression in macrophages can be induced by ox-LDL in a dose-dependent and time-dependent manner.RIP2 gene silencing can inhibit ox-LDL induced M1 macrophage transformation.

Aim To observe the effects of receptor interacting protein 2(RIP2)on macrophage inflammatory ac-tivation and polarity transformation,and to explore the mechanism of RIP2 in macrophage phagocytosis of oxidized low den-sity lipoprotein(ox-LDL).Methods THP-1 derived macrophages were treated with different doses(10,25 and 50 mg/L)of ox-LDL for 24 hours,and treated with 50 mg/L ox-LDL for 8,16 and 24 hours.Real-time quantitative PCR and Western blot were used to detect the expression of RIP2 mRNA and protein in THP-1 derived macrophages,and ELISA was used to detect the secretion of tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein-1(MCP-1).Three pairs of RIP2 siRNA were designed,transfecting them into cells using hiperfict transfection reagent,real-time quanti-tative PCR and Western blot were used to detect the expression of RIP2 mRNA and protein in THP-1 derived macrophages after transfection,in order to screen for the optimal siRNA transfection concentration and the most effective pair of siRNA.After transfection with the most effective RIP2 siRNA,cells were treated with 50 mg/L ox-LDL for 24 hours,ELISA was used to detect the secretion of TNF-α,MCP-1,interleukin-10(IL-10)and interleukin-12(IL-12),real-time quantitative PCR was used to detect the expression of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1),flow cytometry was used to detect the expression of cell surface antigens CD86,CD80 and CD163.Results Ox-LDL induced the ex-pression of RIP2 in macrophages in a dose-dependent and time-dependent manner.With the increase of ox-LDL treatment dose and time,the expression of RIP2 mRNA and protein increased.Among them,the expression of RIP2 protein in the 50 mg/L group was 7.6 times of the control group,and the expression of RIP2 protein in the 24 h group was 17.9 times of the control group(P＜0.001).The ELISA results showed that with the increase of ox-LDL treatment dose and time,the secretion of TNF-α and MCP-1 increased(P＜0.05).After transfection of RIP2 siRNA into cells,ELISA results showed that the secretion of TNF-α,MCP-1 and IL-10 in the ox-LDL group was 2.4 times,2.9 times and 1.8 times of the control group(P＜0.01),and the secretion of IL-12 decreased by 34.2％compared to the control group(P＜0.01);the secretion of TNF-α,MCP-1 and IL-10 in the siRNA+ox-LDL group decreased by 37.4％,45.3％and 27.4％,respectively,com-pared to the ox-LDL group(P＜0.01),while the secretion of IL-12 increased by 31.4％(P＜0.05).The results of flow cytometry and real-time quantitative PCR showed that the expression of CD86,CD80 and iNOS mRNA in the ox-LDL group was 14.2 times,33.8 times and 4.5 times of those of the control group,respectively,while the expression of CD163 and Arg-1 mRNA decreased by 33.4％and 43.0％,respectively,compared with the control group(P＜0.05);the expression of CD86,CD80 and iNOS mRNA in the siRNA+ox-LDL group decreased by 27.6％,29.3％and 34.3％,respectively,compared to the ox-LDL group,while the expression of CD163 and Arg-1 mRNA increased by 30.3％and 38.6％,respec-tively(P＜0.05).Conclusion RIP2 expression in macrophages can be induced by ox-LDL in a dose-dependent and time-dependent manner.RIP2 gene silencing can inhibit ox-LDL induced M1 macrophage transformation.

Objective To investigate the impact of 5-fluorouracil (5-FU) and cisplatin on miR-449b expression in human hepatocellular carcinoma (HCC) and elucidate the molecular mechanism of 5-FU and cisplatin inhibiting the migration of HCC cells.Methods Real-time qPCR analysis was conducted to determine the expression of miR-449b in 50 HCC tissues.RT-PCR assay was performed to detect the expression of miR-449b in HCC cells with 5-FU and cisplatin treatment.The migration of HCC cells with the overexpression of miR-449b was determined by wound-healing assay;Rescue assay was employed to investigate the correlation between 5-FU & cisplatin, miR-449b and the migration capacity of HCC cells;The putative targets of miR-449b were predicted and validated using target prediction programs and immunoblots.Results The expression of miR-449b decreased in HCC tissues (P<0.0001).miR-449b expression increased in HCC cells upon the treatment of 5-FU and cisplatin (P<0.001).The overexpression of miR-449b inhibited the migration of HCC cells (P<0.001).Rescue assay revealed that inhibition of miR-449b to prevent 5-FU and cisplatin induction resulted in suppressed migration in SMMC7721 cells(P<0.05).Catenin-δ was a functional target of miR-449b.Conclusions 5-FU and cisplatin inhibit the migration of HCC cells at least partly via inducing the expression of miR-449b.

Objective To explore the correlation between serum growth differentiation factor‐15(GDF‐15) level and acute myocardial infarction(AMI) to provide a basis for the prognostic evaluation of AMI .Methods Totally 192 Han patients with AMI (AMI group) and non‐coronary heart disease (NCHD ,NCHD group) diagnosed in Chengde Municipal Central Hospital from Sep‐tember 2013 to January 2015 ,were selected and their clinical data were collected .The biochemical markers and serum GDF‐15 level were detected .Results Comparing the AMI group with the NCHD group ,differences in the patients′age ,smoking ,blood glucose (Glu) ,TC ,TG ,LDL‐C levels had statistical significance (P<0 .05);the serum GDF‐15 level in the AMI group was significantly higher than that in the NCHD ;serum GDF‐15 level was positively correlated with TC ,LDL‐C ,hs‐CRP and Glu in the AMI group . Conclusion The increase of serum GDF‐15 level is obviously correlated AMI ,therefore GDF‐15 can serve as an indicator for moni‐toring myocardial infarction .

ObjectiveTo assess the diagnosis ability of contrast-enhanced ultrasound (CEUS)between qualitative analysis and quantitative analysis in breast masses.Methods The contrast-enhanced ultrasound (CEUS) imagings of 73 cases breast masses (41malignant cases,32 benign cases) were analysed retrospectively,including qualitative indexes(obtained by eyes-watching method) and quantitative indexes (obtained by time-intensity curve).The relationship between these CEUS signs and pathology were evaluated by single variety and multiple variety analysis.The two logistic models on the basis of CEUS signs were obtained.According to the logistic regression results to create new variants pre-1 and pre-2,the two receiver operating characteristic (ROC) curves were created to assess the performance of the logistic models.ResultsBy qualitative analysis,the lesion enhanced features,lesion diameter expanding or not and the grades of lesion enhancement at the peak time entered the logistic regression.And the prediction accuracy to differential diagnosis the breast masses was 91.8％ and the area under the ROC was 91.3％.However,by quantitative analysis,only the relative peak intensity entered the regression,the prediction accuracy was 61.5 ％ and the area under the ROC was 75.7 ％.According to Z test,the areas difference under the two ROCs had statistically significant ( P ＜ 0.05).Conclusions To differential diagnosis the breast masses by CEUS,the qualitative indexes have more useful than the quantitative indexes.

OBJECTIVETo prepare effervescent osmotic pump tablet (EOPTs) according to the rhythm of coronary heart disease based on efficacy material and the mechanism of compound Danshen and to study the mechanism of drug released of that tablets.

METHODSince compound Danshen consist of compounds with polyphenolic groups or carboxyl groups, such as phenolic acids, flavonoids, and triterpenoids that they were acidic. EOPTs were prepared from tablet cores which containing NaHCO3 as effervescent, NaCL and manitol as osmotic agents, HPMC as retarding agents coating with CA membrane. And study the mechanism of drug released according to the change of tablet osmotic pressure.

RESULTThe results of in vitro experiments showed that no difference was observed among the profiles of Danshensu, protocatechuic aldehyde, ginsenoside Rg1, Rb1, notoginsenoside R1 release EOPTs. The drug was completely released from the device with a zero-order release rate over 12 h.

CONCLUSIONEOPTs are Successfully obtained EOPT which the drug is released from the device over 12 h and the release mechanism of EOPTs is explained.

Coronary Disease ; physiopathology ; Drug Compounding ; Drugs, Chinese Herbal ; administration & dosage ; metabolism ; Infusion Pumps ; Osmosis ; Salvia miltiorrhiza ; metabolism ; Tablets ; Time Factors

The abilities of self-study,thinking and communication are much more emphasized in the problem based learning(PBL)than in the conventional curriculum.From the view of the inherent relationship between the ability training and PBL,this paper probes into the characteristics of PBL to find the way of fitting the situation of our country.It is essential to the successful teaching and training more high quality students.

AIM: To investigate the regulatory effects of lipopolysaccharide binding protein(LBP)on activation of p38 signaling pathway induced by lipopolysaccharide(LPS)in alveolar macrophages. METHODS: The LBP from actue phase rat serum was purified by ammonium sulphate precipitation, Bio-Rex70 resin and the MonoQ column. Rat alveolar macrophages were exposed to LPS (0 01 mg/L or 1 mg/L) the various concentrations of LBP(0 mg/L, 0 01 mg/L, 0 1 mg/L,1 mg/L and 10 mg/L) Western blotting were used to detect phospho-p38 in alveolar macrophages RESULTS: SDS-PAGE analysis indicated that the purified preparation of rat LBP showed homogeneity and the molecular weight was 60 kD.The binding of lipopolysaccharide to mononuclear cells were enhanced by purified rat LBP. Stimulation of rat alveolar macrophages with LPS at concentration of 0.01 mg/L was LBP dependent. LBP at concentrations up to 1 mg/L was able to increase the activation of p38. However , when LBP concentrations were further increased to 10 mg/L, the phosphorylation levers of p38 were lower as compared with that in the presence of 1 mg/L. Stimulation of rat alveolar macrophages with LPS at concentrations of 1 mg/L was LBP-independent. CONCLUSION: The activation of p38 induced by LPS at lower concentration(0.01 mg/L ) was LBP-dependent, meanwhile, LPS at higher concentration (1 mg/L ) was LBP-independent.