Objective To evaluate clinical curative effect of levosimendan therapy on patients with refractory heart failure.Methods A total of 84 patients with refractory heart failure were randomly and equally divided into le-vosimendan group and routine treatment group.Both groups received routine antiheart failure medication,levosimendan group received levosimendan therapy while routine treatment received milrinone injection therapy additionally.Changes of left ventricular ejection fraction(LVEF)and plasma level of N terminal pro type B natriuretic peptide(NT -proB-NP)were compared between two groups before and after treatment.Results Compared with routine treatment group, there were significant increase in total effective rate of LVEF[(0.36 ±0.18)% vs.(0.42 ±0.36)%],and in NT -proBNP[(975.14 ±247.01)ng/mL vs.(832.14 ±224.78)ng/mL].The effect before and after treatment of levosi-mendan group were more obviously (NT -proBNP:t =2.3 -230.2,P <0.02;LVEF:t =2.29 -215.2,P <0.01). Conclusion Levosimendan can significantly improve heart function,decrease NT -proBNP level in patients with re-fractory heart failure.

Object To determine and compare the content of polysaccharides in three species of Semen Plantaginis from different places of origin. Methods Spectrophotometry was used. Results The content of polysaccharides in the seed of Plantago major L. was higher than those in the seed of P. asiatica L. and P. depressa Willd. Conclusion The content of polysaccharides in Semen Plantaginis of different species or origins are obviously different.

Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.

OBJECTIVE:To study the preparative methods of the long-circulating solid lipid nanoparticles(LSLN)carrying ginkgolides A and B(GAB)and to study the physicochemical characteristics of the GAB-LSLN.METHODS:GAB-LSLN was prepared by ultrasonication or high pressure homogenization.Transmission electron microscopy was employed to study its shape.Particle size,zeta potential,and entrapment efficiency of GAB-LSLN were determined,and its stability after storage under room temperature for 4 weeks was determined as well.RESULTS:The GAB-LSLN prepared by ultrasonication was platelet-shaped and irregular,and that prepared by high pressure homogenization was spherical and regular in shape.The particle diameters of GAB-L SLN prepared by ultrasonication and high pressure homogenization were(219.6?14.3)nm and(173.9?10.4)nm respectively(P0.05).CONCLUSION:High pressure homogenization is superior to ultrasonication in that the prepared GAB-LSLN has small particle size,high stability and high entrapment efficiency.

AIM:To clarify the mechanism of treating autoimmune cardiomyopathy at different stages with anti-L3T4 monoclonal antibody.METHODS:Mice immunized with human mitochondria ADP/ATP peptides were used as the cardiomyopathy(DCM) group,and the sham-immunized mice were regarded as the controls.Mice receiving early treatment were immunized with the same peptides,followed by the injection of 400 ?g of anti-L3T4 on day 0,1 and 2 post-immunization.Mice in the late treatment group were immunized as of the early treatment group but anti-L3T4 was administered 3 months post-immunization.The cytokine expression was measured with three-color flow cytometry to quantitate the splenic Th1/Th2 cell subsets in the different groups of mice.In addition,serum and myocardial cytokines were measured by enzyme-linked immunosorbent assay and real-time PCR.RESULTS:Th1 and Th2 subsets in the early treatment group were similar to those in control group,but were drastically lower than those in DCM group.Mice in the late treatment group showed an increased level of Th1-related cytokines,while the Th2 level was between the DCM and early treatment group.IFN-? and IL-6 levels in early treatment group were similar to those in control group.In the early treatment group,IL-4 level was higher than that in control and lower than that in DCM group,whereas IL-2 and TNF-? contents were lower than those in control and DCM group.In the late treatment group,IFN-? and IL-2 levels were higher than those in DCM group and lower than those in the early treatment group,while IL-6 and IL-4 levels were lower than those in DCM group.CONCLUSION:These results suggest that the cytokine production in cardiomyopathic mice may be repressed by treatment with anti-L3T4 at different stages.Early treatment with anti-L3T4 has better inhibitory function than treatment in late stage of autoimmune cardiomyopathy.

AIM:To explore the effect of recombinamt rat CC16 protein (rCC16) on LPS-induced expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 in the rat tracheal epithelial (RTE) cells. METHODS:The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS (0.1 mg/L) treatment for further 24 h.The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR.The cell culture supernatants were collected for analyzing the protein levels of TNF -α, IL-6 and IL-8 by ELISA.In addition, the nuclear translocation of nuclear factor-κB (NF-κB) p65 was tested by Western blot.RESULTS:rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent (0~2 mg/L) manner, as demonstrated by RT-qPCR and ELISA.However, no concentra-tion-dependent manner between the dose of rCC 16 and TNF-αexpression was observed , and rCC16 inhibited LPS-induced TNF-αexpression at lower concentration (0.5 mg/L).rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION:rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 pro-duction through inactivation of NF-κB activity in RTE cells .

The objective of this study is to develop a sensitive and reliable high-performance liquid chromatography mass spectrometry (LC-MS) method for simultaneous determination of artemisinin, arteannuin B, artemisic acid, and scopoletin, and study the pharmacokinetics of the four constituents in mouse serum after oral administration of the four components to mice. The analytical column used was Agilent Zorbax SB-C18 (2.1 mm x 150 mm, 5 mm). The mobile phase was acetonitrile: 0.5% acetic acid (60: 40) and the flow rate was 0.3 mL x min(-1). The temperature of the column was 40.0 degrees C. In this condition, we established an analysis method to simultaneously determine the four components. A sensitive and specific liquid chromatography-mass spectrometric (LC-MS) method was developed and validated for the determination of artemisin in derivatives in mice plasma. The method we established has a linear range of 5-3 000 μg x L(-1) with a good sensitivity and specificity for all of the four components. This method is simple, rapid, accurate and suitable for the determination of the content of the four compounds.
Animals ; Artemisinins ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Dose-Response Relationship, Drug ; Male ; Mice ; Reproducibility of Results ; Scopoletin ; blood ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; methods

Objective To investigate the protective effect of methylene blue (MB) on blood-brain barrier (BBB) injury after focal cere-bral ischemia-reperfusion in rats. Methods 18 male Sprague-Dawley rats were randomly divided into sham-operated group (n=6), model group (n=6) and MB treatment group (n=6). The left middle cerebral arteries were occluded for 1 hour and reperfused. MB was infused intra-venously immediately after reperfusion (3 mg/kg) and again 2 hours post-reperfusion (1.5 mg/kg), while normal saline was administered in the model group. The sham-operated group was treated as same as the model group without occlusion and infusion. HE staining was used to observe the histological injury in the cortex around the infarcted region 47 hours after reperfusion, while albumin immunohistochemistry was used to evaluate the permeability of the BBB, and immunohistochemistry and double immunofluorescence staining were used to exam-ine the expressions of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4). Results HE staining showed that cells and blood ves-sels were not intact in the cortex around the infarcted region in the model group and they were better in the MB treatment group. The expres-sions of the albumin, GFAP and AQP-4 were higher in the model group than in the sham-operated group (P<0.01), and were lower in MB treatment group than in the model group (P<0.05). The double immunofluorescence staining showed the colocalization of GFAP and AQP-4 in the astrocytes. Conclusion MB may ameliorate the BBB disruption induced by focal cerebral ischemia-reperfusion through reducing glio-cyte proliferation and down-regulation of AQP-4 expression in rats.

Objective To transfect EGFP gene to porcine embryonic fibroblasts ( PEFs) of Tibetan miniature pigs by Lonza Nucleofector II machine and compare the tansfection efficiency between this method and the lipofection method. Method A plasmid carrying green fluorescent protein ( GFP) was transfected into PEFs of Tibetan miniature pigs via the Lonza Nucleofector II machine ( program U020) and by Lipofectamine 2000.Results 5 hours after nucleofection, green fluorescence was observed, indicating 80%transfecting efficiency in the nucleofection group, which is significantly higher than the lipofection group. Conclusion Nucleofector II machine can efficiently transfect PEFs, provides a reliable method for efficiently generate transgenic Tibetan minipigs.