Objective:To investigate the stiffness characteristic of triple-negative human breast cancer at different size in a implantation nude female mice model using shear wave elastography(SWE) and to compare the clinical pathologic features of tumors with elasticity variables.Methods:Human breast cancer MDA-MB-231 cells were injected into 30 nude female mice and 27 transplanted tumors were successfully found in nude female mice. Ultrasound and SWE were longitudinally performed on maximum diameter plane of 21 tumours in 21 nude mice. The elastic parameters of maximal elasticity(Emax), mean elasticity (Emean) and standard deviation of elasticity(Esd) were recorded. The mice were divided into 3 groups according to the tumor size. They were group A with tumor size less than or equal to 5 mm, group B with tumor size greater than 5 mm and less than or equal to 10 mm, group C with tumor size larger than 10 mm and smaller than or equal to 15 mm. Compared with pathology, the relationships between Ki67 of transplanted tumor and elastic parameters were analyzed.Results:As the transplanted tumors increased, the values of Emax, Esd, Ki67 all increased. The lesions maximal size, Emax, Esd, Ki67 were significant higher in group B ( P<0.001, P=0.006, P=0.002, P=0.026) and group C ( P<0.001, P<0.001, P<0.001, P=0.028) than group A. The other parameters were not significantly different among the groups(all P>0.05). The size of transplanted tumors was significantly and positively correlated with Emax ( rs=0.673, P=0.001), Esd ( rs=0.661, P=0.001), and Ki67 ( rs=0.509, P=0.018). Conclusions:SWE Emax and Esd can reflect the tumor tissue stiffness change and biological activity during the tumor growth.

Objective:To study the correlation between echogenic foci pattern of papillary thyroid carcinoma(PTC) solitary nodule and lateral cervical lymph node metastasis.Methods:The clinical data of 475 patients with echogenic foci in preoperative ultrasound and pathologically confirmed PTC solitary nodule from January to December 2014 in Fudan University Shanghai Cancer Center were retrospectively analyzed, which was categorized into lateral cervical lymph node metastasis group and lateral cervical lymph node non-metastasis group. Echogenic foci was classified into five types: local punctate echogenic foci, diffused punctate echogenic foci, coarse echogenic foci, mixed echogenic foci, peripheral annular and eggshell echogenic foci. The related clinical characteristics and the ultrasonic features were also involved. Chi-Square test and Logistic regression analysis were performed to analyze the correlation.Results:Age, preoperative human thyroglobulin (HTG) level, maximum diameter, location and echogenic foci pattern distributed to lateral cervical lymph node metastasis of PTC solitary nodule in 475 cases, according to univariate analysis ( P<0.05). Multivariate Logistic regression analysis showed diffused punctate echogenic foci, age≤35 years old, maximum diameter >10 mm and upper part were risk factors of lateral cervical lymph node metastasis of PTC solitary nodule. Conclusions:Diffused punctate echogenic foci in PTC solitary nodule, patients younger than 35 years old, maximum diameter larger than 1 cm and nodule location at upper part of the thyroid promote to lateral cervical lymph node metastasis.

Objective To compare the difference of diagnostic ability between ultrasound radiomics ( USR) and different conventional imaging models of central neck ( Ⅵ ) lymph node metastasis in papillary thyroid carcinoma ( PTC) . Methods A training set of 609 cases was set up . USR features were extracted and screened by USR method . A weighted formula was established to calculate the USR score of each patient by ultrasound image . T he USR score of the best diagnostic ability was obtained by statistical method and set as the diagnostic criterion . A test set of 326 cases was established to compare the diagnostic ability of USR score with ultrasound ( US ) ,computed tomography ( CT ) and US combined CT . Results T he accuracy ,sensitivity ,specificity ,area under ROC curve and Youden index of USR score in test set were 0 .804 ,0 .867 ,0 .770 ,0 .766 ,0 .533 ,respectively ,which were significantly higher than the corresponding values of US ,CT and US combined CT ( all P ＝ 0 .000 ) . Conclusions USR score obtained with USR method can effectively predict lymph node metastasis in Ⅵ region of PTC . T he diagnostic efficiency and clinical value of USR score were significantly higher than those of conventional medical imaging models .

Objective: To compare the difference of diagnostic ability between ultrasound radiomics (USR) and different conventional imaging models of central neck (Ⅵ) lymph node metastasis in papillary thyroid carcinoma (PTC). Methods: A training set of 609 cases was set up. USR features were extracted and screened by USR method. A weighted formula was established to calculate the USR score of each patient by ultrasound image. The USR score of the best diagnostic ability was obtained by statistical method and set as the diagnostic criterion. A test set of 326 cases was established to compare the diagnostic ability of USR score with ultrasound (US), computed tomography (CT) and US combined CT. Results: The accuracy, sensitivity, specificity, area under ROC curve and Youden index of USR score in test set were 0.804, 0.867, 0.770, 0.766, 0.533, respectively, which were significantly higher than the corresponding values of US, CT and US combined CT(all P=0.000). Conclusions USR score obtained with USR method can effectively predict lymph node metastasis in Ⅵ region of PTC. The diagnostic efficiency and clinical value of USR score were significantly higher than those of conventional medical imaging models.

Objective To investigate the application value of tuberculosis infection of T cells spot test in the diagnosis for pulmonary tuberculosis in elderly patients with chronic obstructive pulmonary disease(COPD).Methods According to test different ways,64 elderly COPD patients with pulmonary tuberculosis were divided into the two groups.The control group(n =32)was checked by tuberculosis antibody test,and the study group(n =32)received T cells infected with tuberculosis spot test.The two sets of results were observed and compared.Results The study group by T cell spot test TB infection,29 cases were detected positive(90.63%),3 cases of negative(9.37%);tuberculosis antibody detection by the control group,23 cases were detected positive(71.88%),9 negative patients (28.12%).Compared the two groups,the study group had higher positive rate(χ2 =3.954 7,P <0.05),and nega-tive rate was lower(χ2 =3.914 7,P <0.05).Conclusion Tuberculosis infection T cell spot test in the diagnosis of pulmonary tuberculosis in patients with COPD in the elderly has greater value,more accurate diagnosis,reliability and worthy of promotion.

Objective To investigate the role of additional post core biopsy ultrasound in clinically node negative breast cancer.Methods Axillary ultrasound was performed before and after breast cancer was diagnosed on core biopsy samples.Post core biopsy ultrasound were performed by radiologists of this department of ultrasound at random.Post-diagnosis ultrasounds were performed by a radiologist with over 20 years of experience for the diagnosis of breast cancer with axi[lary disease.Results were compared to the final axillary pathological result.Results Of the 96 patients,17 were pathology lymph node positive.Post biopsy ultrasound identified 8 of the 17 positive nodes,with a sensitivity 47.1％,specificity 88.6％,positive predictive value of 47.1％,negative predictive value of 88.6％,accuracy of 81.3％.While the diagnosis index of pre-biopsy ultrasound were 47.1 ％,88.6％,47.1％,88.6％,81.3％,respectively.Conclusions Post-biopsy ultrasounds had an increased sensitivity for identifying positive axillary nodes,at the same times,specificity decreased.

Objective To compare the therapeutic effects of continuous veno-venous hemofiltration (CVVH) versus repeated intermittent veno-venous hemofiltration (RIVVH) on patients with severe acute pancreatitis (SAP).Methods Fifty-six patients with SAP were randomly divided into the CVVH group (n =28) and the RIVVH group (n =28).The clinical symptoms and signs,the APACHE Ⅱ and MODS scores,the result of biochemistry including amylase and lipase,and the plasma levels of TNF-α,IL-6,IL8 before and after treatment,the duration of mechanical ventilation,boosting drug application time,the length of stay in ICU,the surgical intervention rate and the mortality were compared between the two groups.Results The clinical symptoms improved in the two groups after treatment (P ＜ 0.05).The APACHE Ⅱ and MODS scores were all reduced in the two groups after treatment (P ＜ 0.05).When compared with the RIVVH group,the result of biochemistry including amylase and lipase,and the plasma levels of TNF-α,IL6,IL-8 were significantly decreased (P ＜ 0.05).The duration of mechanical ventilation,the length of stay in ICU and the mortality were also significantly decreased in the CVVH group (P ＜ 0.05).Conclusions CVVH was more efficacious than RIVVH in the treatment of SAP.

To establish a gene regulation system compatible with biopharmaceutical industry and gene therapy, we constructed a fusion protein of biotin ligase from Bacillus subtilis (BS-BirA) and the trans-activation domain, and used its expression vector as the regulatory vector. Meanwhile, BS-BirA-specific operators were ligated upstream of attenuated CMV promoter to obtain the response vector. In this way, a novel eukaryotic gene regulation system responsive to biotin was established and named BS-Biotin-On system. BS-Biotin-On system was further investigated with the enhancing green fluorescent protein (EGFP) as the reporter gene. The results showed that our system was superior to the current similar regulation system in its higher induction ratio, and that the expression of interest gene could be tuned in a rapid and efficient manner by changing the biotin concentrations in the cultures, Our results show that the established system may provide a new alternative for the exogenous gene modulation.
Bacillus subtilis ; Biotin ; chemistry ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation ; Genetic Vectors ; Promoter Regions, Genetic ; Trans-Activators

Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Currently, exogenous gene expression system based on retroviral vector has been widely used as efficient gene expression system in both gene therapeutic research and RNA interference. In this study, we evaluated the efficiency of exogenous gene expression mediated by the retroviral vector in mammalian cells. First, we constructed EGFP (enhanced green fluorescent protein) vector using pcDNA3.1(+) and retroviral vector pQCXIN as backbone vector respectively. Then, we transfected or infected HEK293 cells and CHO-K1 cells with above vector or corresponding retroviral virus, and measured the relative fluorescence intensity (RFI) of EGFP. The results showed that the RFI of the retroviral virus-infected cells was two times higher than that of the plasmid-transfected cells. Further experiments revealed repeated virus infection enhanced the expression of EGFP markedly, with RFI increasing twice after four rounds of virus infection. Furthermore, the EGFP expression in HEK293 cells mediated by the retroviral vector was more stable than transfected with plasmid pcDNA3.1(+). Finally, we further validated the efficiency of exogenous gene expression system based on the retroviral vector by expressing recombinant human activated protein C (rhAPC) in HEK293 cells. We obtained HEK293 cell lines with rhAPC expression between 10 and 15 microg/(10(6) cells d). In conclusion, the exogenous gene expression system based on the retroviral vector is an alternative method for the generation of stable and high-expressing mammalian cell lines.
Animals ; CHO Cells ; Cricetinae ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Protein C ; biosynthesis ; genetics ; RNA Interference ; Recombinant Proteins ; biosynthesis ; genetics ; Retroviridae ; genetics ; metabolism ; Transfection