The technique of lentivirus-based RNAi ( RNA interference) is a combination of the effective infec-tion and integrating characteristics of lentivirus vector and the specificity of RNAi to silence a homologous gene expression. It caused the post-transcriptional gene silencing ( PTGS) by a specific target sequence of mRNA degradation.As a kind of commonly used target cells in gene therapy, T lymphocytes play an important role in graft-versus-host disease ( GVHD) and other diseases.It is of great significance for increasing the infection efficiency of the target gene to T lymphocytes on this kind of disease for gene therapy.In this paper, we summarize the application of lentivirus-based RNAi in T lymphocytes.

In this experiment, we introduced human interleukin-2(lL-2)gene and Neomycin resistance (Neo~R) gene into a human lung adenocarcinoma cell line GLC, using a retroviral vector PLXSN. The positive cells obtained by selecting in G418 were compared with the wild-type cells. We observed their growth characteristics in vitro and the quantity of IL-2 in the culture supernatants. PCR result demonstrated the integration of foreign genes into tumor genome DNA after transfection and permanent existence. Under electron microscope, what can be seen was the surface villi of trans-duced-cells are less and shorter. It deserves further investigation to determine whether the change of cell surface influence metastatic ability of cells.

Objective To assess the effect of aortic valve replacement (AVR) on left ventricular (LV) twist function in severe aortic stenosis (AS) patients with preserved LV ejection fraction (LVEF) by speckle tracking imaging (STI).Methods Twenty-eight severe aortic stenosis (AS) patients (17 male,age 64.1 ± 10.4 years) with normal LVEF (≥50％) were examined by STI before and six months after AVR.The standard mitral valve and apical short-axis views were obtained to analyze LV basal and apical peak systolic rotation.LV peak systolic twist was calculated as the different between apical rotation and basal rotation.LV fractional shortening (LVFS) was calculated as the percentage fall of LV systolic dimension with respect to diastolic dimension.The data were compared with 28 age and sex-matched normal controls.Results In patients,LVEF remained unchanged after AVR.LV twist increased before (19.7° ± 5.7° vs 12.9° ± 3.2°,P ＜0.001) because of increased apical rotation (13.0° ± 5.8° vs 7.6° ± 2.6°,P ＜0.001),and normalized after AVR (14.4°± 5.2°,P ＜0.001).In controls,LV twist correlated with LVFS (r =0.81,P ＜ 0.001),this relationship was reversed in patients before (r =0.52,P ＜0.01) and after AVR (r =0.34,P ＞0.05).Conclusions In patients with severe AS and normal LVEF,LV twist is exaggerated suggesting potential compensation for reduced long axis function.These disturbances normalize within six months of AVR but lose their relationship with basal LV function.

Objective To access the left internal thoracic artery (LITA) graft late postoperative patency after coronary artery bypass grafting (CABG) by peripheral blood vessel ultrasound combined with color Doppler coronary flow imaging (CDCFI).In contrast with angiography,try to find available flow parameter to access graft patency.MethodsForty-six patients with CABG more than 1 year postoperatively followed-by angiography were detected by ultrasound.The LITA graft and left anterior descending artery were examined.Systolic and diastolic peak velocity(Smax,Dmax),velocity time integral(VTIs,VTId)of each segment were measured separately.The ratio of diastolic and systolic peak velocity (D/S),and diastolic velocity time integral fraction(DVTIF) were calculated.All patients were divided into groups according to angiography results.ResultsThirty -one LITA grafts were patent,11 were dysfunctional,4 were occlusive.According to the angiography results,the flow parameters of the proximal segment of LITA graft were significant.The D/S and DVTIF of patent group was higher than that of dysfunctional group.The diastolic peak velocity of distal segment of LAD of patent group was higher than that of dysfunctional group.ConclusionsPeripheral blood vessel ultrasound combined with CDCFI could provide the evidence to access the patency of the graft.It was an effective method for the clinical follow-up.

OBJECTIVE:To establish a method for the determination of content and content uniformity in Bisacodyl enter-ic-coated tablet. METHODS:HPLC method was performed on the column of Agilent ZORBAX C18 with mobile phase of acetoni-trile-20 mmol/L ammonium acetate(adjusted pH to 5.0 with acetic acid)(55∶45,V/V),the detection wavelength was 265 nm,col-umn temperature was 30℃,flow rate was 1.0 ml/min,and the volume injection was 20 μl. RESULTS:The linear range of bisaco-dyl was 50-1 000 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 99.50%-101.17%(RSD=0.5%,n=9). CONCLUSIONS:The method is reproducible with high accuracy,and suitable for the quali-ty control of Bisacodyl enteric-coated tablet.

Background and purpose:Gene fusions have been identiifed as recurrent oncogenic events in lung adenocarcinoma. Our purpose are to study the histologic features of anaplastic lymphoma kinase (ALK), c-ros oncogene 1 receptor tyrosine kinase (ROS1) andRETproto-oncogene fusion-positive lung adenocarcinomas and to evaluate the correlation between psammoma bodies and fusion-positive lung adenocarcinomas.Methods:In this study, we performed a comprehensive histologic analysis of 44 fusion-positive (including 15RET, 20ALK and 9ROS1)
lung adenocarcinomas and 111 fusion-negative [including 20 epidermal growth factor receptor (EGFR), 20 Kirsten rat sarcoma viral oncogene (K-ras), 71 pan-negative] lung adenocarcinomas.Results:ALK,RET andROS1 fusion-positive lung adenocarcinomas were more prevalent in solid or acinar predominant adenocarcinoma. Multivariate analysis showed that tumors harboring a fusion gene had significantly higher prevalence of the presence of signet ring cells (P=0.000), micropapillary component (P=0.044), mucinous cribriform pattern (P=0.000) and extracellular mucin (P=0.010). The incidence of psammoma bodies was higher in the lung adenocarcinomas with a gene fusion than in tumors without gene fusions (P=0.000). Psammoma bodies were more likely to be found in tumors with any micropapillary component and/or mucinous cribriform pattern than in tumors lacking a micropapillary component and/or mucinous cribriform pattern (P=0.000).Conclusion:Our data showed that the presence of psammoma bodies, micropapillary component, mucinous cribriform pattern, extracellular mucin or signet ring cells may be either sensitive or speciifc to predict tumors harboring a fusion gene. These distinct morphologic features may be helpful in selecting cases for further accurate molecular testing.

Objective To develop a computer-auto-controlling and analysis system for Tail suspension .Methods Combining the advantage of computer science , engineering and animal behavior into tail suspension test .The “energy”index was developed .The system was validated with antidepressants such as imipramine and paroxetine .Results The measuring principle is based on the energy developed by mice trying to escape from their suspension .During the test, the movements of the mice are analyzed in terms of force , energy and power developed over time .Each mouse issuspended by the tail using adhesive tape to a hook connected to a tail test sensor .The tail test sensor fixed to suspension bar picks up all movements of the mouse and transmits these to a signal regulation unit and transmission circuit , which amplify , filter and digitalizes the signals.The signals are displayed visually in waveform .Activity time, immobility time, energy are continuously updated .The correlation coefficient of “immobility time” collected by computer and manpower was 0.94. Using this system, both imipramine and paroxetine could decreased the immobility time and paroxetine could increased the energy induced by mice ( both P <0.01 ) .Conclusion A stable computer-auto-controlling and analysis system for Tail suspension was established and could be used to screen the antidepressants .

The key factor of chemotherapy for brain disorders is to penetrate brain blood barrier(BBB). Receptor mediated en-docytosis is one of the mechanisms for drug carriers to cross the BBB. Angiopep-2(ANG)can specifically bind to low density lipopro-tein receptor-related proteins 1(LRP-1)overexpressed on BBB and glioma cells,thus exhibits a higher LRP-mediated BBB penetra-tion capability. Its targeting efficiency is higher than that of other targeting molecules such as transferrin and(TF)lactoferrin(LF). This paper introduces the structure and function of ANG,and its application in brain targeting delivery systems,in order to provide references for research and development of brain targeting preparations and guidance for therapy of brain diseases.

Aim To study the inhibitory effect of NO donating-aspirin derivative Ⅱ_6 on thrombosis and its mechanism of nitric oxide release. Methods Inferior vena cava thrombosis and cerebral thrombosis model in rats were used to investigate the antithrombotic activities of Ⅱ_6.TXB_2 and 6-Keto-PGF_(1?)in rat serum、rat aortic strip and in ECV304 cells were determined by radioimmunosay.NO content in the serum of rats and extracellular fluid of the ECV304 cells were tested with Griess reagent.Results Ⅱ_6 inhibited the thrombosis in inferior vena cava and brain,and it reduced the concentration of TXB_2.but increased NO production.Conclusion Ⅱ_6 has an inhibitory effect on thrombosis and increases NO content in vivo and vitro.

Objective To investigate the feasibility of VTQ value of spleen in noninvasive assessment of staging of liver fibrosis in patients with hepatitis B.Methods VTQ values of spleen were measured in 56 patients with hepatitis B by using ARFI technique and liver biopsy was performed.The patients were then grouped according to the stages of liver fibrosis stage.The results were analyzed by the intergroup comparison,Pearson Correlation analysis,and receiver operating characteristic curve (ROC curve).Results There was positive association of the VTQ value with the stages of liver fibrosis,whose correlation coefficient was 0.83 (P ＜ 0.05);There was no significant difference between the spleen VTQ value and the liver fibrosis S1 and SO (P ＞ 0.05),whereas there were significant differences among other groups (P ＜ 0.05).The area under the ROC curve was 0.95,the cut-off value was 2.79m/s,and the specificity and sensitivity were 81.8％ and 91.7％ respectively.Conclusions The VTQ value measured by ARFI technique has a better value in noninvasive diagnosis of stages of hepatitis B liver fibrosis.