Objective To investigate whether ultrasound-mediated microbubble destruction(UMD) could enhance cationic liposome (CL) induced plasmid DNA delivery or not,and optimize the transfection conditions.Methods Multiple parameters were explored to obtain the optimal transgene efficiency by means of with or without serum in culture medium,various CL or nano-liposomal bubble(NB) concentrations,different time point of ultrasonic irradiation.The transfection efficiency was assessed by fluorescence microscopy and flow cytometer,and cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay.Results The serum could protect the cells but show little impact on transfection efficiency induced by CL.CL and plasmid DNA at a weight ratio of 4 ∶ 1 exhibited high transfection efficiency of (17.71-± 0.79)％ and high cell viability of (91.28 ± 0.76) ％.CL combining with ultrasonic irradiation at the time point of 1 hour could increase the transfection efficiency to (24.85 ± 0.78)％ (P ＜0.01).Higher transfection rate (32.47 ± 4.01) ％ was obtained by adding NB at the concentration of 10 ％ (P ＜0.05).Conclusions UMD accompanied with CL could enhance gene delivery effectively,which would provide a new method for gene therapy.

Objective To observe self-made cationic nanobubbles as non-viral gene carrier to transfer green fluorescent protein reporter gene into HepG2 cell in vitro.Methods Cationic nanobubbles(PNB) were prepared by sonicating liposomes、polyethylenimine and perfluoropropan.The surface potential and the size of nanobubbles were assessed by laser particle analyzer.HepG2 cells were incubated with DNA,nanobubbles with or without ultrasound exposure.The transfection efficiency was evaluated by flow cytometer and the cell viability by cell counting Kit-8.Results The mean diameter of PNB was (834.57 ± 6.4) nm and the surface charge was (4.15± 0.98)mV.The PNB-DNA complexes,which blocked by the Agarose gel electrophoresis,could effectively transfer HepG2 cells,and the ultrasound exposure could enhance the transfection efficiency further significantly (P ＜ 0.05).Conclusions The new PNB could effectively combine with pDNA to enhance gene delivery and ultrasound exposure could improve its efficiency further in HepG2 cell in vitro.

Objective To made a kind of nano-liposomal bubbles(NB) and observe its base physical properties and ultrasound contrast enhanced effect,and compared with SonoVue in vitro and in vivo.Methods The liposome was made with the method of reverse phase evaporation,and then the liposome was sonieated to prepare NB.After the physical properties of NB such as its morphous,average particle size,surface potential and concentration were observed and calculated,200 μl of saline and two contrast agents (NB and SonoVue) were injected into degassed PBS (10 ml) respectively to observe their enhancing effects.For in vivo study,a healthy rabbit heart and liver were imaged before and after intravenous injection of 2.0 ml/kg of saline and two contrast agents in succession to compare their dynamic enhancing effects in the grey scale imaging subjectively.Results The self-made NB distributed uniformly and its size ranged from 133.1 ～199.5 nm with a mean diameter of (171.60 ± 30.82) nm.Its surface potential and concentration were -(1.92± 0.65)mV and (3.8 ～ 5.6) × 108/ml individually.These basic characteristics were not observed changing dramatically after placement one week to one month under room temperature.The NB and SonoVue all displayed significantly enhancing effect comparing with saline no matter in vitro or in vivo.Conclusions The self-made NB are stable and distribute uniformly,which display the same contrast enhancing effect as SonoVue in vitro and in vivo.The self made NB would have a more potential use in ultrasound molecular imaging and gone or drug delivery.

Objective: To investigate the value of ultrasound-guided fine-needle aspiration (FNA) biopsy combined with thyroglobulin in fine-needle aspirate fluid (FNA-Tg) measurement in diagnosis of preoperative lateral neck lymph node (LN) metastasis of papillary thyroid carcinoma (PTC). Methods: Totally 111 preoperative PTC patients diagnosed with FNA with suspected metastatic LNs were collected and then underwent neck ultrasonography together with FNA cytclogy. FNA-Tg was measured, then the eluents were obtained by flushing aspiration needle with 1 ml normal saline. FNA-Tg of elution was measured by electrochemiluminescence immunoassay. Taken postoperative pathologic results as the gold standards, ROC curve was drawn, the optimal threshold value for diagnosis of FNA-Tg, the diagnostic efficiency of FNA, FNA-Tg, combined FNA and FNA-Tg for suspected metastatic LNs were evaluated. Results: Among 118 suspicious LNs, 81 (81/118, 68.64%) LNs of metastasis and 37 (37/118, 31.36%) LNs without metastasis were confirmed by postoperative pathology. The average value of FNA-Tg in metastasis LNs was significantly higher than that of LNs without metastasis (P<0.001). The optimal diagnostic value of FNA-Tg in diagnosis of LNs metastasis was 2.65 μg/L, and the AUC was 0.937 (95%CI [0.894, 0.980], P<0.001). The differences of accuracy, sensitivity and negative predictive value were all statistically significant between FNA-Tg and FNA (all P<0.05). The accuracy, sensitivity and negative predictive value of FNA combined with FNA-Tg were higher than that of only FNA (all P<0.05). Twenty-two FNA false-negative nodes were correctly diagnosed with FNA-Tg testing. Conclusion: FNA cytology and FNA-Tg detection are useful ancillary tests that can improve the detection of lateral neck LN metastasis of PTC. Combination of FNA and FNA-Tg can significantly increase the accuracy and sensitivity of preoperative diagnosis of lateral neck LN metastasis of PTC, and may have an important role in surgical planning of PTC patients.