Objective: Based on tumor angiogenesis theory, a target system of tumor endothelial cells was established to investigate the cytotoxicity against tumor vascular endothelium by an drug loaded immunoliposome.Methods: Sterically stabilized liposomes containing doxorubicin were used for conjugation to a monoclonal antibody (MAb) as a drug-vascular targeting device. The MAb M5/114 directly against MHC Ⅱ, which was expressed by vascular endothelial cells was employed. Binding of cholesteryl ether (CHE) labeled anti-MHC Ⅱ immunoliposomes to endothelial cells (SVEC ) in vitro was detected by a binding assay. Cytotoxicity of drug loaded immunoliposome to endothelial cells was determined by MTT assay.Results: At least 3-fold increase in binding for the targeted liposomes was observed. This data suggest highly selective binding of M5/114-immunoliposomes to MHC Ⅱ expressing endothelial cells. Moreover, the cytotoxicity produced by drug loaded M5/114-immunoliposome was greatly enhanced compared with that by non-targeted liposomes. Conclusion: This is the first report using immunoliposome to target endothelial cells. The results demonstrate that immunoliposome directly against tumor vascular endothelial cells might provide a general treatment for solid tumors.

Objective To investigate the differential diagnostic value of CT scan minimum attenuation values (minAVsCT) in adrenal adenomas and non-adenomas.Methods CT scan data of 89 cases of clinical and pathologically confirmed adrenal adenomas were subjected to retrospective analysis and compared with data of 46 cases involving 50 non-adenomas (25 metastases,20 pheochromocytomas,3 lymphomas,and 2 cortical carcinomas).The distributions of mean attenuation values (meanAVs) ￥ 10 Hu and minAVs ￥0 Hu and CT histogram analysis with ≥ 10％ negative pixels were observed in adrenal adenomas and non-adenomas,and the diagnostic sensitivity and specificity of these 3 methods for adenomas were calculated.Results The distributions of unenhanced meanAVs ￥ 10 Hu,minAVs ￥0 Hu,and CT histogram analysis with ≥ 10％ negative pixels among cases of adenoma and non-adenoma were 62.9％ (56/89) and 0％ (0/50) (x2=52.687,P=0.000),84.3％ (75/89) and 2％ (1/50) （x2=83.917,P=0.000),and 77.5％ (69/89) and 0％ (0/50) (x2=83.917,P=0.000),respectively.The respective diagnostic sensitivities,specificities,false negative rate (FNR),false positive rate (FPR),positive pre dictive value (PPV),negative predictive value (NPV) and accuracy of these 3 methods for adenomas were 62.9％vs 84.3％ vs 77.5％,100％ vs 98.0％ vs 100％,37.1％ vs 15.7％ vs 22.5％,0 vs 2％ vs 0,100％ vs 98.7％ vs 100％,60.2％ vs 77.8％ vs 71.4％,and 76.3％ vs 89.2％ vs 85.6％.Conclusion Although the specificity and PPV of minAVs≤0Hu is slightly less than meanAVs≤10Hu and CT histogram analysis with ≥ 10％ negative pixels,it exhibits the best sensitivity and accuracy with a simple operation,and is thus suitable for clinical application.

Objective To investigate the diagnostic value of hyperenhancement sign on ultrasound,CT and their combination in diagnosis of thyroid benign and malignant nodules.Methods The contrast-enhanced ultrasound and enhanced CT datas of 172 thyroid nodules in 144 cases confirmed by operation and pathology were retrospectively analyzed,including 97 benign nodules and 75 malignant nodules.According to the degree of enhancement,these nodules were divided into iso-or low-enhancement and hyperenhancement.The distributions of ultrasound hyperenhancement,CT hyperenhancement and both of them in thyroid benign and malignant nodules,adenomatoid lesions and nodular goiters were analyzed,followed byx2 test for statistical analysis.Results In 172 thyroid nodules,the proportions of ultrasound hyperenhancement,CT hyperenhancement and both of them in thyroid benign and malignant nodules were 53.6％(52/97) and 20.0％(15/75)(x2＝20.090,P＜0.05),34.0％ (33/97) and 4.0％ (3/75) (x2＝23.033,P＜0.05),31.0％ (30/97) and 0％ (0/75) (x2＝28.096,P＜0.05),respectively.Their sensitivity and specificity of diagnosing benign nodules were 53.6％ (52/97) and 80.0％ (60/75),34.0％ (33/97) and 96％ (72/75),30.9％ (30/97) and 100％ (75/75),respectively.The proportions of ultrasound hyperenhancement,CT hyperenhancement and both of them in adenomatoid lesions and nodular goiters were 96.9％(31/32) and 32.3％(21/65)0x2＝35.946,P＜0.05),65.6％(21/32) and 18.5％(12/65)(x2＝21.250,P＜0.05),65.6％(21/32) and 13.8％ (9/65)(x2＝26.912,P＜0.05).Their sensitivity and specificity of diagnosing adenomatoid lesions were 96.9％(31/32) and 67.7％(44/65),65.6％(21/32) and 81.5％(53/65),65.6％(21/32) and 87.2％(56/65).Conclusions Ultrasound hyperenhancement,CT hyperenhancement for diagnosing thyroid benign nodules are of significant value.Especially for adenomatoid nodules,ultrasound hyperenhancement has a higher sensitivity,while CT hyperenhancement has a higher specificity.Their combination can further improve the diagnostic specificity,thus reducing the unnecessary surgical trauma.

Objective To discuss the predictive value of multiple ultrasonic features combination in diagnosis of thyroid papillary carcinoma (PTC) more than 1.0 cm in diameter. Methods The ultrasonic features of 258 PTC nodules from 251 patients and 207 nodular goiter (NG) nodules from 190 patients in the First People′s Hospital of Hangzhou were retrospectively analyzed. All the nodules were confirmed by pathological examination after surgery. The ultrasonic features included the shape of nodules, internal echo,anteroposterior/transverse diameter ratio (A/T), and microcalcification. The χ2 test was used to analyze the differences of ultrasonic features between PTC and NG. Multi-variate analyses (Logistic regression) was used to analyze the predictive risk ultrasonic features of PTC. The sensitivity and specificity of ultrasonic features were analyzed based on the gold standard of pathological results. Results There were significantly differences between 258 PTC nodules and 207 NG nodules in irregular shape, hypoechogenicity,A/T > 1 and microcalcification (χ2 values were 121.511, 105.411, 41.483, 121.072, all P < 0.01). The results of Logistic regression showed that irregular shape, hypoechogenicity, A/T>1 and microcalcification were risk ultrasonic features of PTC. And their OR values were 5.013 (95%CI 2.919-8.610), 5.811 (95%CI 3.411-9.901), 15.399 (95%CI 7.576-31.301), 4.141 (95%CI 1.687-10.164) respectively. The sensitivity and specificity of single ultrasonic feature were 26.0%-79.5% and 71.5%-96.1%; the sensitivity and specificity of two ultrasonographic features combination were 11.2%-57.0% and 92.3%-99.0%; and the sensitivity and specificity of three or four ultrasonographic features combination were 8.1%-31.8% and 99.0%-99.5%.Conclusions Irregular shape, hypoechogenicity, A/T> 1 and microcalcification of thyroid neoplasm are important ultrasonic features of PTC. Although the sensitivity of single ultrasonic feature in diagnosing PTC is higher than that of multiple features combination, it has a lower specificity. Therefore, combination of multiple ultrasonographic features can improve the specificity in diagnosing PTC and reduce the misdiagnosis of PTC.

OBJECTIVE The aim of the study was to evaluated the difference and consistency in tumor size measured by sonographic and pathological examination in papillary thyroid carcinoma(PTC).METHODS A total of 114 patients with PTC, including 122 malignant nodules, was collected from Hangzhou First People's Hospital between Jun 2012 and Jun 2014. The tumor sizes were measured by preoperative sonographic and postoperative pathologic evaluation. Pearson correlation analysis, paired t-test, and Bland-Altman plot were used to evaluate the correlation and consistency in tumor size measured by the two methods.RESULTS Pearson correlation analysis showed that the largest tumor size measured by sonography were positively correlated with pathologic size (r=0.957, P=0.000). Paired t-test showed that there were statistically difference between sonographic size and pathological size (8.24±5.06) mmvs (7.79±4.75) mm,P=0.001. The absolute difference value of the largest tumor size measured by the two methods was from zero to 6.5 mm, with the average of (1.03±1.14) mm. Bland-Altman analysis showed that the limits of agreement (LoA) of difference was from -2.41 mm to 3.33 mm, with the 95% confidence interval from -2.87 mm to 3.78 mm.Within the limit of the consistency, the maximum moduli was 2.9 mm.CONCLUSION There is a significant discrepancy between the preoperative sonographic and the pathologic size of the papillary thyroid carcinoma, which should be taken into account in clinical practice.

Objective Based on contrasting CT signs distributed in PTC greater than 1.0 cm in diameter and nodular goiters(NG),multiple logistic regression analysis is adopted to make a statistics of what diagnostic value that CT signs have for PTC.Methods Retrospective analysis of CT findings of 288 PTC with diameter ＞1.0 cm in 277 cases confirmed by histology,including nodular form,cookie bite symptom,microcalcifications and en hanced narrowing/blurring was performed,and compared with CT signs of 276 NG in 231 cases with diameter＞1.0 cm.Analysis of various CT signs were performed by multivariate Logistic regression method,and the sensitivity,specificity of positive CT signs and their combinations in PTC diagnosis were calculated.Results Multivariate Logistic regression analysis showed that irregular nodules,cookie bite symptom,microcalcifications and enhanced arrowing/hlurring were often observed in PTC,with OR values of 17.249(95％ CI 8.954-33.227),23.697 (95％ CI 11.653-48.188),4.536 (95％ CI 2.031-10.132),4.672 (95％ CI 8.954-8.999),respectively.The sensitivity,specificity of single CT sign diagnosing PTC were 31.3％-82.3％ and 83.3％-93.1％,respectively.The sensitivity,specificity of two CT signs combinations diagnosing PTC were 24.0％-70.5％ and 96.7％-100％,respectively.The sensitivity,and specificity of three or four CT signs combinations diagnosing PTC were 19.1％-61.5％ and 99.6％ -100％,respectively.Conclusions Although the nodule form,cookie bite symptom,microcalcifications and enhanced narrowing/blurring are the important signs for diagnosing PTC,the OR values of various signs had great difference.The accurate identification of these differences and various CT signs combinations can further improve the specificity of diagnosing PTC,thus reducing misdiagnosis.

Objective:To investigate the expression and distribution of human dermal papillary fibroblasts (Fp) , reticular fibroblasts (Fr) , and myofibroblasts (MFB) in keloid tissues.Methods:Keloid tissues were collected from 15 outpatients (including 8 males and 7 females) aged 20-50 years, who were diagnosed in the Department of Dermatology, Renmin Hospital of Wuhan University from May to December 2019. Normal skin tissues were taken from 15 age-matched women who underwent mammoplasty, and served as controls. The distribution of fibroblast activation protein (FAP) , CD90 and alpha-smooth muscle actin (α-SMA) was observed in the keloid tissues and normal skin tissues by dual immunofluorescence staining. Furthermore, fibroblasts were isolated from 3 normal skin and 3 keloid tissue samples, and subjected to primary culture. Subsequently, the fibroblasts were treated with 10 ng/ml transforming growth factor-β1 (TGF-β1) for 48 hours in vitro, during which, changes in fibroblast phenotypes were observed in the 2 groups. Fluorescence-based quantitative RT-PCR and Western blot analysis were performed to determine the mRNA and protein expression of FAP, CD90 and α-SMA. Measurement data were compared between 2 groups by using t test. Results:Immunofluorescence staining of the normal skin tissues revealed that FAP +/CD90 - fibroblasts were predominantly distributed in the superficial dermis, FAP -/CD90 + fibroblasts in the deep dermis, and CD90 + cells hardly expressed α-SMA; however, a large number of FAP + fibroblasts and CD90 + fibroblasts were observed in the deep keloid tissues, and many CD90 + fibroblasts also expressed α-SMA. Dual immunofluorescence staining showed that normal tissue-derived fibroblasts hardly expressed α-SMA, and keloid-derived fibroblasts expressed α-SMA. The fluorescence intensity of α-SMA + cells significantly increased in the normal tissue-and keloid-derived fibroblasts after 24-hour treatment with TGF-β1 (21.058 ± 0.709, 27.112 ± 0.097, respectively) compared with that in the corresponding untreated fibroblasts (11.312 ± 0.636, 21.306 ± 0.464, t=22.430, 13.370, respectively, both P < 0.05) . RT-PCR and Western blot analysis showed that the mRNA and protein expression of FAP, CD90 and α-SMA significantly increased in the keloid-derived fibroblasts after 48-hour treatment with TGF-β1 (mRNA: 92.610 ± 3.667, 1.366 ± 0.105, 3.240 ± 0.141; protein: 0.652 ± 0.073, 1.046 ± 0.119, 0.946 ± 0.117, respectively) compared with the untreated keloid-derived fibroblasts (all P < 0.05) . Conclusion:CD90 + Fr aberrantly proliferated in the deep dermis of keloid tissues, suggesting that directional intervention in aberrantly proliferating FAP -/CD90 + Fr in the deep dermis may promote the efficacy for keloids.

Objective:To explore the predictive value of various ultrasonic signs for papillary thyroid microcarcinoma (PTMC) .Methods:The ultrasonic data of of 603 micronodular goiter (MNG) in 396 cases and 640 PTMC in 539 cases, which were confirmed by pathology from Jan. 2013 to Dec. 2016, were retrospectively analyzed. According to the different inspection time, all nodules were divided into model group (2013-2014 years) and test group (2015-2016 years) . The tumor morphology, internal echo, microcalcification, and aspect ratio (A/T) were observed. Chi-square test and multivariate Logistic regression analysis were used to analyze the distribution differences of the four ultrasound features in PTMC and MNG, and their diagnostic value was evaluated.Results:There were statistical difference between model group and test group in ultrasonic signs including tumors shape, internal echo, microcalcification and aspect ratio according single factor analysis (chi square value was 283.540 and 298.119, 63.130 and 87.400, 26.342 and 50.152, 169.918 and 181.405; P<0.05) ;Multivariable Logistic regression analysis showed that irregular shape, hypoecho, A/T>1 and microcalcification were more common in PTMC ( P< 0.05) . OR values were 18.410 and 19.231, 2.560 and 6.380, 9.379 and 6.724, 3.102 and 8.830, and AUC prediction probability values were 0.916 and 0.911 respectively. Conclusions:Irregular shape, internal hypoechoic, microcalcification and A/T>1 are stable important ultrasonographic signs in predicting PTMC. Comprehensive analysis of various ultrasonic signs can improve the diagnostic efficiency.

Objective To explore the value of ultrasound gray scale ratio (UGSR) in the diagnosis and differential diagnosis of papillary thyroid carcinoma(PTC) with different sizes.Methods A retrospective study was made in 702 patients with 1107 nodules which were confirmed by surgery in the Department of Oncology or fineneedle aspiration of HangZhou First people's Hospital,Zhejiang University of medical school from Jan.2016 to Oct.2017.All the thyroid nodules were divided into three groups:D≤ 1 cm group,1＜D≤2 cm group and ＞2 cm group according to their sizes.The UGSR of the PTC and NG were obtained through the RAD info system.Their differences were analyzed and ROC was established to confirm the optimal threshold in the differential diagnosis between PTC and NG among the groups.Results There were 483 PTC and 624 NG in this study.The UGSR of D≤ 1 cm group,1＜D≤2 cm group and ＞2 cm group of PTC and NG were (0.48±0.12) vs (0.76±0.22)(t=33.21,P=0.00);(0.52±0.17) vs(0.80±0.21)(t=1.30,P=0.00) and (0.63±0.20) vs(0.89±0.24)(t=3.58,P=0.00) respectively.The area under the ROC of UGSR in the differentiation of PTC and NG in the three groups were 0.873,0.840 and 0.811 respectively.The Youden indexes were greatest (0.631,0.536 and 0.535 respectively),when the cut-offs of the UGSR were 0.682,0.652 and 0.831 respectively.The sensitivity and specificity to diagnose PTC were 94.8％ and 68.0％,75.0％ and 78.6％,80.3％ and 73.2％ respectively in the three groups.Conclusions The best UGSR value of PTC was variant in thyroid nodule with different size.Recognition of these differences accurately could improve the pre-operative diagnostic accuracy of PTC.Also the method is simple to operate and easy to apply.