The gene silence induced by small interfering RNA (siR N A) provides a powerful tool for gene therapy of neoplasm. However, safe and efficient delivery of exogenous siRNA is still the key issue for clinical application. Polyethyenimines ( PEI ) and its copolymers can encapsulate siRNA into complex in the formation of nanoparticle wherein the stability and activity of siRNA are closely related to the structure and molecular weight of PEI, N/P ratio, and the composition of copolymers.The siRNA-encapsulated nanoparticles can effectively inhibit the proliferation of tumor cell not only in vitro, but also in vivo by administration via intratumoral,intraperitoneal or venous injections.

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 evaluate the clinical value of ultrasonography guided percutaneous core needle biopsy in pancreatic lesions. Methods Thirty-four patients with 36 pancreatic lesions in Shanghai First People′s Hospital Afifliated to Shanghai Jiao Tong University from February 2012 to November 2013 underwent conventional ultrasound-guided percutaneous core needle biopsy using automatic gun and 18-gauge biopsy needles. The site, size, internal and surrounding vascularity, the sampling number of the lesions, and whether the specimens′ quality was satisfied were recorded. Then specimens were sent for pathological examination, and all above observations were compared with the ifnal diagnosis. Results The number of lesions with 2, 3 and 4 samplings was 32, 2 and 2, respectively. The average number of sampling was 2.2 (mean, 2.17;standard deviation, 0.51) and the acquisition rate of satisifed specimens was 89%(32/36). The pathological results of biopsy were malignant in 31 of 36 lesions including 27 cases of ductal adenocarcinoma, 2 cases of lymphoma, 1 case of small cell neuroendocrine carcinoma and 1 case of uterine leiomyosarcoma metastasis. The other 5 lesions were non-malignant including 3 cases of benign lesion, 1 cases of atypical hyperplasia and 1 cases of granulation tissue. The 36 lesions were ifnally diagnosed as 34 cases of pancreatic malignancy, 2 cases of non-malignant neoplasm. The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of ultrasonography guided percutaneous core needle biopsy in pancreatic lesions were 91%(31/34), 100%(2/2), 92%(33/36), 100%(31/31) and 40%(2/5), respectively. Youden index was 0.91. Two patients had mild upper abdominal pain and 1 patient had transient elevated serum amylase. No pancreatitis, pancreatic fistula, peritonitis, bleeding or dispersion of malignant cells along the penetrating channel or other serious complications occurred. Conclusion Ultrasonography guided percutaneous core needle biopsy is a simple, rapid, safe and effective diagnostic method in pancreatic lesions with high clinical value.

Objective To observe the therapeutic effects of the parenchymal malignancies treated with argon-helium cryoablation percutaneouslly.Methods 49 foci in 45 patients with malignancies were treated with argon-helium cryoablation percutaneouslly.The temperature in the cryoablated area was dynamically recorded.The size and coverage of ice-balls formed were monitored.The imaging changes were observed.The shrinkage of tumor was followed 3 months post-cryoablation.The foci were divided into two groups in size:less than or equal to 3 cm and more than 3 cm.The chi-square test and spearman analysis were used by SPASS 11.0 software.Results The temperature in the cryoablated area descended fast to below-130℃ in a minute and relatively kept maintaince about-150℃ or so in the whole procedure.It ascended to 15℃ in a minute after rewarming.The ice-balls were formed within 50 seconds after cryoablation in 87.5% foci.And it increased in size as the time going.The coverages of the iceball more than or equal to and less than 80% were 72.7%、27.3% and 26.3%、73.7% in groups of less than or equal to 3 cm and more than 3 cm,respectively(P＜0.05).The ice-ball was demonstrated as arc hyperechogenicity with shadow and round-like low density on sonography and CT imaging,respectively.The shrinkage more than 50%of the foci was 81.8% and 30.6% in the groups of less than or equal to and more than 3 cm three months after the cryoabalated procedure(P＜0.01).Conclusion It is an actual/exact and convient way for the pereutaneous argon-helium cryoablation in treating malignancy.It can be used in the management of the parenchymal malignancy whole body.

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 survey the incidence of sleep disorders among patients with industrial injuries and analyze the relevant factors.MethodsA total of 112 depressed patients ( male 106,female 6; aged 22-79 years,course of disease 35 d-25 years) were assessed by a professional psychologist using life satisfaction index A,the type A behavior pattern scale,the Pittsburgh sleep quality index (PSQI) and the Barthel index.ResultsThe incidence of disordered sleep was 40.18％ (45/112),of whom mildly depressed patients were 34.29％,moderately depressed patients 42.86％ and severely depressed patients 85.71％.The incidence of sleep disorders increased with increasing depression severity.The incidence of disordered sleep was significantly higher among the severely depressed patients than among those mildly or moderately depressed,but there was no significant difference in incidence between moderately and mildly depressed patients.PSQI scores among the severely and moderately depressed patients were significantly higher than among those mildly depressed,but there was no statistically significant difference in average PSQI scores between the severely and moderately depressed patients.The sleep disorder group suffered significantly poorer sleep quality and took significantly longer to get to sleep.There were no significant differences in average age,educational level,marital status,social relations,family and social support,gender distribution or course of disease between the two groups.There were,however,significant differences in family income,life satisfaction,character type and disease species between the groups.ConclusionThe incidence of disordered sleep among depressed patients after industrial injury is correlated with the severity of depression,family income,life satisfaction,the type of injury and the patient's character.

Objective To establish an HPLC quantitative method for the content determination of baicalin in Kangdeling capsule .Methods The chromatography column was Agilent Tc-C18-WR (4 .6 mm × 250 mm ,5μm) ,and the column temper-ature was 30 ℃ .The mobile phase consisted of acetonitrile and 0 .5‰ phosphoric acid (26 ∶ 74 ) .The flow rate was 1 .0 ml/min ,and the detection wavelength was 265 nm .Results The retention time of baicalin was about 16 min .The calibra-tion equation was Y=22 114 .67 X -112 836 .7(r=0998 8)with good linearity in the range of 5 .410-108 .2 μg/ml for baicalin . The average recovery was 98 .78% while RSD were 0 .74% .Conclusion This method is simple ,time-saving and accurate which could be used to routine analysis of baicalin in Kangdeling capsule .

We reconstructed the erythromycin macrocyclic lactone (6-deoxyerythronolide B, 6dEB) synthesis pathway in Escherichia coli. We first cloned all the genes needed to synthesize the 6dEB into multi-gene co-expressed vectors. Then using the recognition sequences of isoschizomers Xba I/Spe I of vectors, we assembled the related genes into a series multiple-genes recombinant plasmids pBJ144, pBJ130. The recombinant plasmids pBJ144, pBJ130 were cotransformed into BAP1 to get the recombinant BAP1(pBJ144/pBJ130). SDS-PAGE analysis showed that individual genes were expressed correctly. After inducing at low temperature, adding propionate as substrate, we validated the crude product by mass spectrometry and the 6dEB yield was about 10 mg/L. These results indicated that the synthetic pathway of 6dEB was successfully assembled and reconstructed in Escherichia coli, which will greatly facilitate the reconstruction of whole erythromycin synthesis pathway and finally help to establish a stable research platform for developing of new derivatives of erythromycin and combinatorial biosynthesis of polyketide-type antibiotics.
Anti-Bacterial Agents ; biosynthesis ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Erythromycin ; analogs & derivatives ; biosynthesis ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics

Objective:To investigate the ultrasonographic features and evolution of thyroid shrinking nodules in order to improve the differential diagnosis and management strategy and avoid unnecessary biopsy.Methods:A total of 245 patients with old bleeding of benign thyroid nodules diagnosed via fine needle aspiration cytology (FNAC) from May 2015 to July 2020 in the southern part of the Shanghai General Hospital Affiliated to the Medical College of Shanghai Jiaotong University, including 263 nodules. The sonographic parameters such as size, shape, aspect ratio, echo, edge, boundary, periphery, halo, calcification, posterior echo enhancement and posterior acoustic attenuation of nodules were analyzed retrospectively, and the ultrasonic appearances of nodules were classified; 41 nodules from 40 patients, who did ultrasound examinations more than twice and had both complete ultrasonographic data, were compared and then classified in order to explore the evolution rules of the nodules.Results:Two hundred and sixty-three thyroid shrinking nodules in 245 cases were classified into 4 types: "cystic wall shrinkage sign" type(71.48%, 188/263), "carcinoma-like" type (22.05%, 58/263), "inflammation-like" type(3.04%, 8/263) and "undefined atypical solid nodules" type(3.42%, 9/263). Of the 41 consecutive follow-up nodules, 37 cases showed obvious cystic wall shrinkage sign, and 7 of them developed into "carcinoma-like" type and 6 cases into "inflammation-like" type.Conclusions:The ultrasonographic appearance of thyroid shrinking nodules is a dynamic process, which can be divided into 4 types: "cystic wall shrinkage sign" , "carcinoma-like" , "inflammation-like" and "undefined atypical solid nodules" types. The "cystic wall shrinkage sign" type is typical and common. The "cystic wall shrinkage sign" type can develope into the "carcinoma-like" type or the "inflammation-like" type.