Objective To optimize the transfecfion parameters for transfecting small interfering RNA (siR-NA) to endothelium of mouse angioma (EOMA) cells mediated by lentiviral vector. Methods According to Poly-brene existing or not(8 μg/ml), series of multiplicity of infection (MOI) (1, 5, 10, 15, 20)and the time of infec-tion(4, 6, 12,24 h), EOMA cells were divided into 40 groups. 140 hours after transfection, cells were counted un-der fluorescent microscope to determine the percentage of the cells transfected under each condition. The cell viability was calculated by using Typan Blue. Results The percentage of positive cells was as high as 83% in the group with Polybrene (8 μg/ml), MOI(5) and the time of tranfectian (6 h), and the livability of EOMA in this group was 96%. After that, the transfection efficiency was not increased with the increasing of MOI, the time of transfection, but the mortality of EOMA was increased markedly. Conclusion The transfection efficiency and cell viability are dependent on the Polybrene(8 μg/ml),correct MOI (5)and the time of transfection (6 h).

Objective To evaluate multi-slice spiral CT venography (MSCTV) and digital subtraction venography (DSV) in diagnosing iliac vein compression syndrome (IVCS) and secondary thrombosis. Methods The imaging materials, including MSCTV and DSV performed before and after the thrombolysis therapy, of 38 patients with clinically-suspected IVCS were collected. The inner diameters of the compressed iliac veins were measured on MSCTV images and the compression ratio was calculated. Usingχ2 test, the detection rates of IVCS by MSCTV and DSV were compared. Results Of 38 patients, IVCS was detected by MSCTV in 29, by pretreatment DSV in 20 and by post-treatment DSV in 29. The difference in the detection rate of IVCS between MSCTV and pre-treatment DSV was statistically significant (χ2=4.65, 0.010.05). Conclusion For the diagnosis of IVCS, MSCTV is superior to pre-treatment DSV in the diagnostic accuracy of iliac vein compression syndrome. Therefore, MSCTV should be used as the preferred method of examination.(J Intervent Radiol, 2015, 24:301-305)

Objective To observe the therapeutic effect of acupuncture therapy for the rat model of type 2 diabetes mellitus (T2DM) combined with renal hypertension and try to explore its mechanism. Methods We randomly select 10 Wistar rats as the blank group and 40 rats were used to make the model groups, which were divided into simple diabetes group, simple renal hypertension group, the compound group with electroacupuncture and the compound group without elec?troacupuncture, with 10 rats in each group. After a high fat and sugar diet for 4 weeks, the Wistar rats were given strepto?zotocin i. p. injection to establish models of type 2 diabetes mellitus. Renal hypertension was developed by the“2K1C” im?proved method to make unilateral renal artery ligation?induced renal artery stenosis. Then, electroacupuncture treatment was performed on the rats for 2 weeks except the compound group without electroacupuncture. The changes of values of BP, FBG, Cr, BUN, glycated hemoglobin, renin and Ang II were recorded and analyzed. Results The values of BP, FBG, Cr, BUN, glycated hemoglobin, renin and Ang II in the compound group with electroacupuncture showed a significant re?duction compared with the compound group without electroacupuncture after 2 weeks (P<0?01), but there was no obvious changes in the values of Cr and BUN(P>0?05). Conclusions The blood glucose and blood pressure in the rat model of compound group can be reduced to a normal level with continuous electroacupuncture at bilateral acupoints Zusanli, and it can also be kept at a stable level after single electroacupuncture for 2-3 days. The acupuncture therapy is more suitable for early clinical treatment and can be used in basic research with advantages of economic, safe, no side effect and so on.

Purpose To explore the value of 128-slice dual source CT (DSCT) three-dimensional post-processing techniques in the diagnosis of acute aortic dissection (AAD). Materials and Methods All image data of 116 patients with AAD who underwent conventional and enhanced DSCT scan by dual-energy scanning technology were retrospectively analyzed, and the multi-planar reconstruction (MPR), volume rendering (VR) and maximum intensity projection (MIP) were conducted in the workstation. Taking digital subtraction angiography (DSA) as the diagnostic gold standard, we analyzed the imaging manifestation of the original and 3D-reconstruction images and evaluated the specificity and the sensitivity of diagnostic accuracy and the image quality. Results The diagnostic accuracy of conventional scan was 37.1%(43/116). The display rates of MPR for initial break, intimal flap and true and false lumen were 93.1%, 100.0%and 100.0%, respectively. The display rates of VR and MIP for the initial break were 33.62%and 6.90%, respectively, which was both lower than that of MPR. The display rate of MIP for the true and false lumen was 23.28%. The overall display capability of MPR was significantly better than that of VR and MIP (P<0.01), and the display capability of VR was better than that of MIP (P<0.01). The sensitivity and specificity of enhanced DSCT in the diagnosis of AAD were both 100.0%. Conclusion DSCT has a fast and reliable diagnostic value on AAD. Conventional CT signs should be highly valued in the evaluation of chest or abdominal pain; and a thin layer of MPR and VR should be chosen in the three-dimensional reconstruction;whilst MIP reconstruction may be unnecessary for AAD.

With citric acid as carbon source and different amino acids as nitrogen source,the nitrogen-doped carbon was synthesized by one-step hydrothermal method under the condition of no catalyst.The pre-experiments show that arginine-doped carbon dots(CDs-Arg) with relatively high fluorescence quantum yield (33.25％) were picked out for further study.Furthermore,we studied the physical and chemical properties of CDs-Arg through a series of spectral,electric potential,particle size,X ray and elemental analysis experiments.At the same time,the stabilities of the nanoparticles towards different pH,temperatures,excitation lasers or redox conditions were studied.And the MTr and in vivo distribution experiments were also conducted for the toxicity and metabolism studies.The experimental results showed that the arginine doped carbon dots has high fluorescence efficiency,good stability,and extremely low toxicity.And the water-soluble small particles of CDs-Arg nanoparticles showed that the nanoparticles can be excreted through the glomerulus.These results show that the CDs-Arg Nanoparticles are biocompatible nanoparticles and have potential applications in biological imaging and can serve as a monitor of drug metabolism.

Purpose　To introduce a method to isolate cDNA clones using bacteriophage P1-derived artificial chromosome (PAC) or bacterial artificial chromosome (BAC) as probe for hybridization and try to find some novel genes related to hepatocellular carcinoma.　Methods　PAC 579 (D17S926 locus) and BAC 1529 (D17S1529 locus) in the deletion region of chromosome 17p13.3 in human hepatocellular carcinoma were chosen to screening the human liver cDNA library as probe for hybridization. The isolated positive cDNA clones were partially sequenced, then analyzed by computer comparison and Southern blot.　Results　After three cycles of screening, 78 and 8 candidate positive cDNA clones were isolated using PAC 579 and BAC 1529 probes respectively. Further analysis indicated 18 cDNA clones isolated by PAC 579 probe and 5 cDNA isolated by BAC 1529 probe were potential novel genes related to hepatocellular carcinoma.　Conclusions　The isolation of cDNA clones using PAC and BAC probes is effective and practical.

Objective To identify two suspected Legionella pneumophila (L.pneumophila) strains isolated from environmental water and air conditioning cooling water systems in Guangzhou city.Methods The two strains were identified by their cultural characteristics,biochemical test,Legionella-specific primer PCR identification,PCR-enzymatic digestion analysis,16S rRNA,mip and rpoB gene sequencing analysis.Results The two suspected L.pneumophila isolates were identified as gram-negative bacillus appeared as white colonies on BCYEα-agar after incubation for 48 hours at 36℃.Both isolates were positive for oxidase,gelatinase and hydrolysis of hippurate,and negative for urease activity and nitrate reduction.Their phenotypic characteristics were similar to those of L.pneumophila strains.Results of PCR identification by using Legionella-specific primer were positive.Enzymatic digestion analysis showed that the 226 bp PCR products of two isolates were not digested by Taa Ⅰ.The two strains were classified as Legionella busanensis as indicated by gene sequencing analysis of 16S rRNA,mip and rpoB gene.Conclusion Two L.busanensis strains were first isolated from environmental and air conditioning cooling water systems in China.Due to their biochemical characteristics,L.busanensis strains were commonly misidentified as L.pneumophila,but could be effectively identified by PCR-enzymatic digestion analysis and multiple genes identification.

Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.

Objective Analysis and adscription of volatiles from Guizhi Tang and study on its improvement of the learning and memory in dementia mice induced by scopolamine.Methods The volatile oil from Guizhi Tang(GZT),Guizhi and Shengjiang was extracted using steam distillation method and was analyzed by GC-MS. Morris water maze and step-down test were carried out for obtain the difference of the learning and memory improvement in 40 ICR mice from randomized groups, such as the control group, the model group, the donepezil group (2 mg/kg), the low dose of volatile oil of GZT (5 mg/kg), and the high dose of volatile oil of GZT (20 mg/kg), and ACh, AchE, BchE and chE in serum were detected by ELISA. Results Among 38 identyfied volatile ingredients from GZT, 18(44% in weight) was from Guizhi, and 9 was from Shengjiang. Compared with the model group, the low and high dose of GZT volatile oil significantly increased swimming distance ratio in destination quadrant (26.74% ± 16.42%vs.9.42% ± 8.50%, P<0.05); goal quadrant time scale (43.51% ± 25.12%vs. 14.50% ± 12.23%,P<0.05)) increased significantly than the model group ; the number of errors in the experiment platform (1.63 ± 1.19vs. 0.25 ± 0.46, P<0.05) obviously increased than model group ; platform test in the made errors times (0.57 ± 0.98vs. 4.43 ± 2.4, P<0.05) significantly reduced. The GZT total volatile oil groups significantly reduced cognitive obstacles small rat serum in the cholinester enzyme (chE) (140.90 ± 3.27, 144.79 ± 6.71vs. 134.49 ± 3.36,P<0.05); acetylcholinesterase (AchE) (3.30 ± 1.31, 3.94 ± 0.78 vs.8.52 ± 3.39,P<0.05); butyrylcholinesterase (BchE) (3.22 ± 0.45, 3.66 ± 0.53vs. 7.99 ± 0.79,P<0.05); and acetylcholine (Ach) (4.10 ± 0.38, 3.03 ± 0.25vs.1.72 ± 0.50, P<0.05) significantly increased.Conclusions The GZT volatile oil mainly from Guizhi and Shengjiang can improve the learning and memory ability in dementia mice induced by scopolamine via a cholinergic mechnism.

Objective To observe the effect of early occupational therapy intervention on unilateral spatial neglect caused by stroke.Methods A total of 92 patients with unilateral spatial neglect caused by stroke were ran- domly divided into a treatment group(46 cases)and a control group(46 cases).The cases of the control group were treated by routine clinical treatment plus positioning on the bed,passive joint motion,bridging movement,facilitation techniques,balance training in sitting and standing positions,gait training immediately after the patients' conditions got stable.The patients in the treatment group were treated with activities of daily living(ADL)training in addition to the interventions employed in the control group.Results Effect of the treatment group are superior to that of the control group as reflected by the scores with Fug1-Meyer scale and Barthel Index(P