Objective To explore the adeno-associated virus (AAV)gene transduction and cellular endocytosis mediated by ultrasound combined with microbubbles in two types of cells.Methods HeLa and NIH/3T3 cells were infected by rAAV2-EGFP at a concentration gradient to get the optimal concentrations for enhancement.At these concentrations,HeLa and NIH/3T3 cells were infected by rAAV2-EGFP mediated by ultrasound combined with microbubbles.The gene transduction efficiency were observed and measured by fluorescence microscopy and flow cytometry at 48 h after treatment.The cell viability was tested by CCK-8.The number and distribution of cellular clathrin-coated endocytic pits were observed by confocal fluorescence microscopy and transmission electron microscopy on 45 min after treatment.Results The optimal concentrations for HeLa and NIH/3T3 cells were 2000 v.g./cell and 10000 v.g./cell.Ultrasound combined with microbubbles significantly enhanced the transduction efficiency of rAAV2-EGFP (P <0.01) without significant cell viability decrease (P > 0.05 ).Confocal fluorescence microscopy and transmission electron microscopy demonstrated that clathrin-coated endocytic pits were more obviously increased in ultrasound combined with microbubbles mediated AAV transduction group than AAV transduction group. Conclusions Ultrasound combined with microbubbles can efficiently enhance the gene transduction of AAV,whose cellular transportation depends on cellular endocytosis,in two types of cells.Stimulating cellular endocytosis might be one of the mechanisms of enhanced cellular transportation of AAV mediated by ultrasound combined with microbubbles.

Objective To determine the diagnosis and the prognosis of acute bilineal leukemia(aBLL).Methods The cases who had been morphologicaUy diagnosed as acute leukemia were retrospectively screened.The characteristics of aBLL blasts and the patients'responses to chemotherapy and the survival rate were identified based on the analysis of both their clinical and laboratory data.Results From total of 352 cases screened,11 cases of aBLL were identified.The incidence of aBLL was 3.1%.Among them 4 were male and 7 female.The median age was 38 years old.The median counts of white blood cell at the onset of the disease was 56.3×109/L.There were more My/B aBLL subtype cases than My/T aBLL (8 cases vs.3 cases). Cytogenefie analysis data were available for 7 cages. All were standard karyotypes except 1 who had complex karyotypie change.The complete remission(CR)rate of chemotherapy was 45.5%(5/11)and the response rate(CR+PR)Wag 54.5%(6/11).Only 2 patients achieved CR after the first course of induction chemotherapy.The median of remission duration was 11 months,ranged from 3 to 14 months.The median of survival time was 10 month. ranged from 1 to 23 months. 1 patient died during the induction course.Conclusion aBLL is a rare subtype of acute leukemia.The diagnosis of the disease could be based on the combinational analysis of morphology and immunophenotype study.The patients'responses to the conventional chemotherapy regimens were not satisfied.Therefore,the prognosis of the disease was poor.

Objective To investigate the intracellular delivery of siRNAs through the applications of ultrasound targeted microbubbles destruction(UTMD)and biodegradable nanoparticles carriers.Methods Preparation of nanoparticles with and without RGD sequences,parameters optimization via L16(45)orthogonal design,control experiments in groups of optimization,RGD targeted nanoparticles,non-RGD nanoparticles and blank control, and determinations by inverted fluorescence microscope and flow cytometry were performed.Results The uptake and fluorescence intensity of PC-3 cells in group of RGD targeted nanoparticle was (93.49±1.37)% and 34.28±2.06 respectively,and that in group of optimization was (88.33±1.24)% and 30.59±3.93 respectively(P＞0.05).Whereas the uptake and fluorescence intensity of PC-3 cells in group of non-RGD nanoparticles was(71.24±2.80)% and 18.39±0.90 respectively,and that in group of optimization was (84.78±2.13)% and 27.18±0.91 respectively(P＜0.05).ConclusionsThe applications of UTMD with RGD targted nanoparticles cannot increase the intracellular delivery of siRNAs.

Objective To explore the difference of the contrast-enhanced ultrasound (CEUS) image and quantitative parameters between two different contrast agents Fluorocarbon and SonoVue. Methods The tumor model of colorectal carcinoma in nude mice was established by injecting CT26 cells into the subcutaneous space in hepatic area of 15 nude mice. CEUS was performed with Fluorocarbon and SonoVue on the 14th day after establishment. SonoLiver software was used to analyze the dynamic image quantitatively. The difference of the CEUS image and quantitative parameters between the two contrast agents was analyzed. Results Compared with normal liver parenchyma around the tumor, the tumor ultrasound contrast performance was fast forward and rewind with low enhancement. There was no signiifcant difference between the two kinds of microbubbles not only for CEUS image but also for quantitative parameters [maximum intensity (Imax):49.53%±24.38%vs 45.04%±17.03%, rise time (RT):11.68 s±3.07 s vs 13.76 s±2.92 s, time to peak (TTP):12.76 s±4.12 s vs 15.26 s±3.74 s, T1/2:50.57 s±28.32 s vs 48.75 s±9.85 s, Imax/TTP(V1):4.48±2.82 vs 3.18±1.49, (Imax-INT60)/(60-TTP)(V2):0.67±0.34 vs 0.60±0.20, AUC1:3032.78%±1343.12%vs 3258.77%±1369.84%, AUC2:11647.38%±6183.10%vs 10439.04%±4604.65%, AUC:14680.17%±7469.85%vs 13697.81%±5831.99%, Rate of AUC1:264.25±146.93 vs 222.24±92.16, Rate of AUC2:241.67±119.97 vs 231.97±100.34, all P>0.05). No mouse was dead during the CEUS examination. Conclusion Fluorocarbon had similar CEUS imaging effect and quantitative information compared with SonoVue.

Objective To study the correlation between quantitative parameters of contrast-enhanced ultrasonography(CEUS)and microvessel density(MVD)in hepatocellular carcinoma(HCC),and to investigate the value of CEUS in noninvasively assessing HCC angiogenesis.Methods The patients with 28 HCCs confirmed by pathology were examined by CEUS.A new type quantification software SW-UCS was used to delineate time-intensity curve(TIC)and to obtain quantitative parameters of HCC and hepatic parenchyma in region of interest.The quantitative parameters were compared with MVD,which was counted by immunohistochemistry method using CD34 antibody,and the correlation between them was studied.Results The TICs of HCC showed typical contrast-enhancement pattern of"fast-in and fast-out"comparing with normal hepatic parenchyma.There were significant differences between HCC and hepatic parenchyma in slope of rise time(a3),time to initial peak(TTP),peak intensity(PI),increased signal intensity(ISI)and acceleration time(ACT)(P＜0.05).MVD in HCC(49.36±12.57)was higher than that in hepatic parenchyma(32.46±11.78,P=0.000).For HCC,the PI,ISI and area under curve were positively correlated with MVD,while the slope of decrease to half of peak(a2)was negatively correlated with MVD (P＜0.05).Conclusions The quantitative parameters of CEUS in HCC were correlated with MVD,CEUS can evaluate the angiogenesis of HCC in vivo noninvasively.

Objective To compare the transfection rate and safety between SonoVue + ultrasound and liposome mediated gene transfection to retinal pigment epithelium (RPE) in vitro, and to approach the feasibility of SonoVue and ultrasound enhancing liposome mediated gene transfection to RPE in vitro. Methods RPE were cultured in 24 well dishes with green fluorescent protein plasmid of 2 μg/well. All the cells were divided into 5 groups: control group, ultrasound irradiance group, SonoVue + ultrasound irradiance group(2 W/cm2 and 3 W/cm2),liposome group and liposome + SonoVue + ultrasound irradiance group. There were 10 wells in every group. Gene transfection results were observed under inversion fluorescent microscope. Seventy-two hours later, gene transfer was guantified under flow cytometry. The apoptosis rates of RPE were determined with Annexin Ⅴ-FITC and PI staining by flow cytometry. Results The transfection rate of ultrasound exposure group was (1.06 ± 0.13)%. The transfection rate of SonoVue + ultrasound group was (15.81 ± 1.70)% and (20.86 ± 2.63)%. The transfection rate of liposome group was (30.06 ± 3.49)% ,and the differences between above mentioned sets were significant(P<0.05). The transfection rate of liposome + SonoVue + ultrasound was (44.51 ± 1.28) %,and the differences between above mentioned two sets were significant (P<0.05). Conclusions The transfection rate of liposome is higher than that of SonoVue + ultrasound group. SonoVue and ultrasound can obviously enhance liposome mediated EGFP gene transfection to RPE in vitro.

Purpose To explore the diagnostic value of double exponential model for pelvic lesions using 3.0T MRI for the diagnosis of pelvic lesion. Materials and Methods Fifty patients with pelvic lesions (30 benign cases and 20 malignant cases) underwent MR750-diffusion weighted imaging (DWI) scans, with b values of 0, 50, 300, 600, 800 and 1200 s/mm2, Functool-MADC software was used on AW 451 workstations for data processing, Slow ADC value, Fast ADC value, Standard ADC value, Fraction of fast ADC value were recorded and compared between benign and malignant lesions, and Standard ADC images were fused with axial T2 fat-suppressed images. Results Slow ADC values [(1.83±0.86)×10-3 mm2/s] and Standard ADC values [(1.79±0.78)×10-3 mm2/s] of benign lesions were larger than those of the malignant lesions [Slow ADC values:(1.05±0.31)×10-3 mm2/s;Standard ADC values:(1.13±0.39)×10-3 mm2/s] (t=3.90, 3.51;P<0.01), and the difference of Slow ADC value was largest between benign and malignant lesions. Slow ADC values of both benign and malignant lesions were significantly less than the Fast ADC values [benign:Slow ADC value=(1.83±0.86)×10-3 mm2/s, Fast ADC value=(16.95±8.63)×10-3 mm2/s; malignant: Slow ADC value=(1.05±0.31)×10-3 mm2/s, Fast ADC value=(15.12±9.90)×10-3 mm2/s] (t=-10.40,-6.29;P<0.01). Conclusion Double exponential decay model is capable of differentiating benign and malignant pelvic tumors, thus is of great significance for clinical preoperative diagnosis.

Objective To evaluate the effect of pelvic muscle exercise on quality of sexual life after panhysterectomy surgery.Methods A retrospective review was made on 166 patients with integrated follow-up documents,after their panhysterectomy surgery caused by benign diseases.A normal nursing care was provided to the control group (n=81); while an additional pelvic muscle exercise was performed on the normal care basis,to those in the experimental group (n=85).Finally,a questionnaire on sexual life quality was made at the sixth month,ninth month,and twelfth month respectively,using a repeated measure ANOVA to analyze the data of each dimension and the whole score.Results Significant differences revealed both on the whole score and dimensions of the sexual satisfaction,sexual communication,sexual anxiety and sexual reaction,in different time sites.However,no difference was found in sexual attitude and body image.Besides,a cross interaction was found between time and intervention,where intervention presented different effects.Conclusions Pelvic muscle exercise is recommended in gynecological continuing nursing care to facilitate the recovery of sexual life after panhysterectomy surgery.

Objective To study the effect of gastrodin on arterial blood gas and brain injury of rats under simulated high altitude hypoxia environment. Methods A total of 60 adult healthy male Wistar rats were randomly divided into normal (N) group, hypoxia model (M) group, rhodiola crenulata (RC) group, low dose of gastrodin (GAS-L) group, medium dose of gastrodin (GAS-M) group and high dose of gastrodin (GAS-H) group (10 for each group). The intragastric administration on rats was continued for 7 days timely in each day. Under simulated 8000m altitude using low pressure oxygen cabin, the arterial blood gas of each group were tested, pathological changes of brain tissues were observed and related indexes of brain were detected after 12h hypoxia. Results Comparing with group N, the blood oxygen partial pressure (PO2), value of blood oxygen saturation (SO2), oxygenation index (PO2/FIO2), Na+ concentration (Na+), actual bicarbonate radical (HCO3–) significantly decreased (P<0.01), lactic acid (Lac), hemoglobin concentration (Hb) significantly increased (P<0.01) and pathological damage was inflicted in group M; and contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) in brain tissue significantly increased (P<0.01), content of glutathione(GSH) and activity of glutathione peroxidase (GSH-Px) in brain tissue significantly decreased (P<0.01) in group M. Compared with group M, PO2, SO2 and PO2/FIO2 significantly increased (P<0.01, P<0.05) in group GAS-L; Na+ and HCO3– significantly increased (P<0.01, P<0.05) in three dose groups of GAS; Lac significantly decreased (P<0.01, P<0.05) in group GAS-L and GAS-H. Hb significantly increased (P<0.01) in group GAS-H, a rising trend appeared in group GAS-L but with no statistical significance. Damages of brain tissue were alleviated in group RC and three dose groups of GAS comparing with group M. Compared with group M, MDA significantly decreased (P<0.01) in three dose groups of GAS; there was a decreasing trend of H2O2 but with no statistical significance in three dose groups of GAS; GSH and GSH-Px significantly increased (P<0.01, P<0.05) in three dose groups of GAS. However, three groups of GAS has no dose dependent. Conclusion There was an protective effect of gastrodin on arterial blood gas and brain injury of rats under simulated high altitude hypoxia environment.

AIM:To observe the effect of arsenic trioxide (ATO) on bleomycin-induced pulmonary fibrosis in rats. METHODS:Pulmonary fibrosis was induced in Sprague-Dawley (SD) rats by intratracheal instillation of bleomycin (BLM). The rats in ATO treatment group,steroid treatment group and model group were intraperitoneally injected with ATO,dexamethasone or normal saline (NS),respectively,while the control rats received NS both intratracheally and intraperitoneally. The effects of ATO were evaluated by analyzing the median survival time,hydroxyproline level in the lung,semiquantitative grading of alveolitis and pulmonary fibrosis,and quantitative analysis of the collagen in lung tissue (Masson's trichrome staining). Apoptosis index (AI) of the lung was detected by using the terminal transferase dUTP-digoxygenin nick endlabeling (TUNEL) method. The results of immunohistochemical staining for some cytokines were quantitatively analyzed. RESULTS:ATO (1) prolonged the median survival time of rats with BLM-induced pulmonary fibrosis at some extent; (2) attenuated the alveolitis and pulmonary fibrosis,reduced hydroxyproline level and collagen deposition in the lung tissue; (3) increased the AI of lung tissue at a certain phase; and decreased the levels of transforming growth factor-?1(TGF-?1) and tissue inhibitor of metalloproteinase-1 (TIMP-1),increased the content of in-terferon-? (IFN-?),but did not influence the concentration of matrix metalloproteinase-9 (MMP-9) significantly. CONCLUSION:ATO might attenuate BLM-induced pulmonary fibrosis in rats via increasing the AI in the lung tissue.