Objective:To investigate the value of targeted three-dimensional contrast-enhanced ultrasound（3D-tCEUS）in efficacy evaluation of non-surgical treatments for renal cell carcinoma（RCC）.Methods:Forty nude mice with subcutaneous xenograft tumor model of human RCC（786-O cells）were divided into four groups based on different treatment methods：control，thermal ablation（TA），sunitinib（Suni），and TA+Suni. 3D-tCEUS were performed on days 1，3，7，and 14 post-treatment using self-developed vascular endothelial growth factor receptor 2（VEGFR2）targeted microbubbles. Tumor overall volume（V T）and non-enhanced volume（V N）were measured，from which the volume of the active（enhanced）region was calculated as V A = V T - V N. The tumor total and active area volume was standardized as V T st and V A st（standardized tumor volume = tumor volume after treatment / tumor volume at the beginning of treatment）. Tumor growth curves were plotted and tumor inhibition rates calculated for V T and V A respectively. Quantitative parameters，including the area under the curve（AUC）and the difference in peak intensity before and after burst（dTE），were obtained from the viable tumor enhanced region，and the standardized targeted quantitative parameters were derived by calculating the ratios of parameters at various time points post-treatment to those pre-treatment. The differences in V T st，V A st，AUC and dTE between different treatment groups at different time points were compared. At the end of the experiment，tumor tissues were obtained for immunohistochemical staining to observe the expression of VEGFR2 and CD31 antigens. Results:During the treatment period，no statistically significant differences in weight changes were observed among groups（all P > 0.05）. When V T was taken as the research object，V T st increased across all groups during the treatment period，with the TA group showing the most significant growth，while the TA + Suni group exhibited the smoothest increase in growth curve. When analyzing the tumor enhanced region，tumor growth trend of V A st was different with V T st for all groups；the Suni group showed a slow upward trend，whereas the TA + Suni group showed a continuous decline. Significant differences in tumor inhibition rate originated from V T and V A were noted within the same experimental group（ P < 0.05 for all experimental groups）. One day post-treatment，the AUC and dTE of the TA group were higher than that of the Control group，while the dTE of the TA + Suni group was lower than that of the Control group（all P < 0.05）. By day 3，statistically significant differences in AUC and dTE were observed between each experimental group and Control groups（all P < 0.05）. At day 14，the TA group showed increased AUC and dTE compared with those before treatment，while all other groups，particularly Suni group and TA + Suni group，demonstrated significant reductions（all P < 0.05）. Immunohistochemical results revealed the highest VEGFR2 and CD31 positivity in the TA group，followed by the Control group，while the Suni and TA + Suni groups exhibited lower rates. Conclusions:The combination of TA and targeted therapy effectively induces RCC cells death，demonstrating superior efficacy compared to monotherapy. 3D-tCEUS serves as a accurate and reliable tool for early evaluating the efficacy of non-surgical RCC treatments.

Objective:To investigate the value of targeted three-dimensional contrast-enhanced ultrasound（3D-tCEUS）in efficacy evaluation of non-surgical treatments for renal cell carcinoma（RCC）.Methods:Forty nude mice with subcutaneous xenograft tumor model of human RCC（786-O cells）were divided into four groups based on different treatment methods：control，thermal ablation（TA），sunitinib（Suni），and TA+Suni. 3D-tCEUS were performed on days 1，3，7，and 14 post-treatment using self-developed vascular endothelial growth factor receptor 2（VEGFR2）targeted microbubbles. Tumor overall volume（V T）and non-enhanced volume（V N）were measured，from which the volume of the active（enhanced）region was calculated as V A = V T - V N. The tumor total and active area volume was standardized as V T st and V A st（standardized tumor volume = tumor volume after treatment / tumor volume at the beginning of treatment）. Tumor growth curves were plotted and tumor inhibition rates calculated for V T and V A respectively. Quantitative parameters，including the area under the curve（AUC）and the difference in peak intensity before and after burst（dTE），were obtained from the viable tumor enhanced region，and the standardized targeted quantitative parameters were derived by calculating the ratios of parameters at various time points post-treatment to those pre-treatment. The differences in V T st，V A st，AUC and dTE between different treatment groups at different time points were compared. At the end of the experiment，tumor tissues were obtained for immunohistochemical staining to observe the expression of VEGFR2 and CD31 antigens. Results:During the treatment period，no statistically significant differences in weight changes were observed among groups（all P > 0.05）. When V T was taken as the research object，V T st increased across all groups during the treatment period，with the TA group showing the most significant growth，while the TA + Suni group exhibited the smoothest increase in growth curve. When analyzing the tumor enhanced region，tumor growth trend of V A st was different with V T st for all groups；the Suni group showed a slow upward trend，whereas the TA + Suni group showed a continuous decline. Significant differences in tumor inhibition rate originated from V T and V A were noted within the same experimental group（ P < 0.05 for all experimental groups）. One day post-treatment，the AUC and dTE of the TA group were higher than that of the Control group，while the dTE of the TA + Suni group was lower than that of the Control group（all P < 0.05）. By day 3，statistically significant differences in AUC and dTE were observed between each experimental group and Control groups（all P < 0.05）. At day 14，the TA group showed increased AUC and dTE compared with those before treatment，while all other groups，particularly Suni group and TA + Suni group，demonstrated significant reductions（all P < 0.05）. Immunohistochemical results revealed the highest VEGFR2 and CD31 positivity in the TA group，followed by the Control group，while the Suni and TA + Suni groups exhibited lower rates. Conclusions:The combination of TA and targeted therapy effectively induces RCC cells death，demonstrating superior efficacy compared to monotherapy. 3D-tCEUS serves as a accurate and reliable tool for early evaluating the efficacy of non-surgical RCC treatments.

Objective:To evaluate the ability of vascular endothelial growth factor receptor 2(VEGFR2)/integrin α vβ 3 dual-targeted microubble (MBD) to target angiogenesis of renal cell carcinoma (RCC) in vivo. Methods:Non-targeted microbubble (MBN) USphere LA was employed as a template to prepare single- and dual-targeted microbubbles which could bind VEGFR2 and/or integrin α vβ 3 (MBV and MBI) by the biotin-avidin bridging method. A total of 40 RCC nude mice models were established by subcutaneously injecting 786-O cells.Twenty of the models were all injected with MBN, MBV, MBI and MBD in a random order, and the other 20 models were registered for antibody blocking assays. The results of ultrasound images were used for quantitative analyses, and the following quantitative parameters were obtained: intensity increment (a 1), peak halving speed (a 2), curve rising slope (a 3), perfusion time (t 0), time to peak (TTP), peak intensity (PI), mean transit time (MTT) and area under the curve (AUC) for the first three minutes, peak intensity at 10 s before (P 1) and after (P 2) ultrasound destruction, and the differences of tissue enhancement (dTE) between P 1 and P 2 (dTE=P 1-P 2). All the quantitative parameters of four contrast agents and the antibody blocking assays were compared.Besides, the immunohistochemical assays were performed to evaluate the expression of CD31, VEGFR2 and integrin α vβ 3 in tumor tissues. Results:The differences of parameters of a 1, a 3, t 0, TTP, PI and P 2 among four different microbubbles had no statistical significances (all P>0.05), and all parameters between the two single-targeted contrast agents were not statistically different (all P>0.05). The parameters of AUC, MTT, P 1 and dTE all showed a trend that dual-targeted bubbles > single-targeted bubbles > non-targeted bubbles (all P<0.05). On the contrary, the trend of dual-targeted bubbles < single-targeted bubbles < non-targeted bubbles (all P<0.05) was observed for a 2. In the antibody blocking experiment, a 2 was faster after the antibody injection ( P<0.001), while AUC, MTT, P 1 and dTE were all lower than those before the antibody injection ( P<0.001), and the other parameters were not statistically different before and after the antibody injection (all P>0.05). Immunohistochemical analyses confirmed the high expression of CD31, VEGFR2 and integrin β 3 in tumor tissues. Conclusions:The VEGFR2 and integrin α vβ 3 dual-targeted microbubble has a good potential to target the angiogenesis of human RCC in vivo.

Objective:To explore the capability of vascular endothelial growth factor receptor 2 (VEGFR2)/integrinα vβ 3 dual-targeted microbubbles in assessing the expression level of pro-angiogenic factors during renal cell carcinoma (RCC) growth. Methods:VEGFR2/integrinα vβ 3 dual-targeted microbubbles were prepared by using biotin-avidin linkage method. Twenty subcutaneous RCC xenografts in nude mice were established by subcutaneously injecting 786-O cells and then divided into 2 groups randomly. The targeted contrast-enhanced ultrasound (t-CEUS) examination was performed for all 10 mice in the first group when xenograft tumors were metered from 5 to 10 mm and >10 to 20 mm respectively. And the quantitative parameters of RCC on t-CEUS were longitudinally evaluated during tumor growth. The second group were divided into two subgroups according to xenograft tumors′ diameter, which was 5 to 10 mm and >10 to 20 mm respectively, and underwent t-CEUS examination. Quantitative analysis was performed for all t-CEUS images to obtain the targeted quantitative parameters, which including peak intensity (PI), area under the time-intensity curve (AUC), the differential tissue enhancement (dTE, presenting the difference in PI before (P 1) and after (P 2) the process of Flash). All xenograft tumors in the second group were harvested for immunohistochemical staining to observe the expression of VEGFR2, integrinα vβ 3 and CD31, and their differences in RCC with different tumor sizes. And the correlations between quantitative parameters and VEGFR2, integrinα vβ 3 and CD31 were analyzed. Results:The longitudinal comparison showed that there were statistically significant differences between AUC and dTE of RCC with different tumor sizes (all P<0.001). The larger the tumor size, the smaller the parameters were. According to the horizontal comparison, the expression levels of VEGFR2 and integrinα vβ 3 in larger RCCs were higher than those of RCCs with smaller size (both P<0.05), but there was no significant difference in CD31 expression between the two subgroups ( P=0.754). Both the targeted quantitative parameters (AUC anddTE ) and pro-angiogenic factors (VEGFR2 and integrinα vβ 3) were negatively correlated with tumor size ( rs=-0.83, -0.81, -0.70, -0.88; all P<0.05). Further more, there were good positive correlations between AUC and VEGFR2, integrinαvβ ( rs=0.76, 0.72; all P<0.05). There were good positive correlations between dTE and VEGFR2, integrinα vβ 3 ( rs=0.81, 0.70; all P<0.05). Additionally, the parameter PI was positively correlated with the expression of CD31 ( rs=0.70, P=0.025). Conclusions:The t-CEUS, mediated by VEGFR/integrinα vβ 3 dual-targeted microbubbles, allows noninvasive assessment of the expression levels of VEGFR2 and integrinα vβ 3 in RCCs, which decrease gradually with the increase of tumor size.

Objective:To investigate the effects of perioperative whole course thermal insulation strategy on extubation, recovery time, coagulation index and the incidence of complications in patients undergoing thoracoscopic surgery.Methods:A total of 121 patients who underwent thoracoscopic surgery who received treatment in Zhoushan Hospital from October 2016 to February 2018 were included in this study. Among them, 59 patients who underwent thoracoscopic surgery from October 2016 to February 2017 were included in the simple thermal insulation group, and 62 patients who underwent thoracoscopic surgery from October 2017 to February 2018 were included in the whole process thermal insulation group. Before and 24 hours and 72 hours after surgery, platelet count, prothrombin time, activated partial thromboplastin time and thrombin time were compared between the two groups. Postoperative extubation time and recovery time were recorded. The incidence of shivering, restlessness and other complications was analyzed.Results:Postoperative extubation time and recovery time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(8.06 ± 4.60) min vs. (13.98 ± 7.22) min, (47.19 ± 12.97) min vs. (56.84 ± 17.49) min, t = 5.40, 3.47, both P < 0.05). At 24 and 72 hours after surgery, platelet count and activated partial prothrombin time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(12.55 ± 0.88) s vs. (13.11 ± 0.97) s, (27.44 ± 2.43) s vs. (29.03 ± 2.14) s, (11.42 ± 0.73) s vs. (11.87 ± 0.74) s, (27.44 ± 1.96) s vs. (28.80 ± 2.22) s, t = 3.32, 3.81, all P < 0.05). The incidence of postoperative shivering and restlessness in the whole process thermal insulation group was significantly lower than that in the simple insulation Group (7 cases vs. 27 cases, 5 cases vs. 22 cases, χ2 = 17.782, 14.894, 3.33, 3.57, all P < 0.05). Conclusion:Perioperative whole course thermal insulation strategy can shorten extubation and recovery time, inhibit perioperative coagulation dysfunction, and decrease the incidence of perioperative restlessness, shivering and other complications caused by hypothermia. The effect of perioperative whole course thermal insulation is superior to that of simple thermal insulation.

Objective:To investigate the value of contrast-enhanced ultrasonography (CEUS) in detecting minute renal cell carcinoma (MRCC) smaller than 15 mm (by ultrasonic measurement) and the strategy to improve its detection rate.Methods:Fifty-three pathologically confirmed MRCCs by surgery from November 2007 to October 2019 at Zhongshan Hospital of Fudan University were enrolled in this retrospectively study. All of them underwent both conventional ultrasound and CEUS examinations. The clinical and imaging data were collected and analyzed. Common features, such as tumor size, location, echogenicity, morphology, border, and blood flow signals were observed on conventional ultrasound. On CEUS, the presence of enhancement, wash in and wash out pattern, perfusion uniformity within the lesions were observed.Results:Post-operative pathology confirmed 48 clear cell carcinomas, 4 papillary carcinomas, and 1 chromophobe cell carcinoma. On conventional ultrasound, 12/53 lesions showed no protrusion out of the kidney, and 41/53 cases slightly protruded out of the kidney. There was considerable difficulty in the detection of ten lesions, which was achieved with the guidance of CT/MRI, due to their dorsal location of the kidney. On conventional ultrasound, solid, hyper-echoic, color flow signal with varying degrees were the main features of MRCC.The boundary could be well- or ill-defined, and cystic changes existed in part of cases. On CEUS, most MRCCs showed simultaneous enhancement in cortical phase, iso- to hyper-enhancement at peak, and rapid washout in parenchymal phase. The comparisons of imaging features demonstrated that the characteristics were significantly different between conventional ultrasound and CEUS with regard to boundaries, blood supply, and perfusion uniformity (χ 2=12.425, 20.247, 7.185; all P<0.01). Conclusions:CEUS can significantly improve the detection rate of MRCC, which is superior to conventional ultrasound.

Objective To compare the ultrasound differences between gouty arthritis with chronic kidney disease( GCKD) and simple gouty arthritis( GA) ,and evaluate the correlation between the number of tophi and the degree of renal impairment . Methods Twenty-two patients with GCKD and 22 patients only with GA were examined by high frequency and color Doppler ultrasound . The differences between the tophus , intra-tendinous aggregate , double contour sign , hyperechoic spots in synovium , synovium hyperplasia ,effusion and erosion of the knee ,ankle and first metatarsophalangeal joint ( M TP 1 ) were examined . The correlation between the degree of renal impairment and the number of tophi was analyzed . Results The prevalence of tophi in GCKD group were significant serious compared with simple GA group ( Z = - 3 .915 ,P < 0 .001) . The articulations involved tophi in GCKD group were more than those in simple GA group( χ2 = 20 .283 , P < 0 .001) . And the tophi in GCKD group were more likely to involve multiple articulations while simple GA group frequently manifested none or single articulation involved( χ2 = 13 .165 , P = 0 .001 ) . The tophus in GCKD group was more frequently involved tendon than that in simple GA group ( χ2 = 6 .783 , P = 0 .009) ,but the number of aggregates in tendon showed no significant difference between the two groups ( Z = - 1 .499 , P = 0 .134) . The double contour sign was more frequent in GCKD group than that in simple GA group( χ2 = 7 .511 , P = 0 .006) ,there was no difference of hyperechoic spots in synovium ,synovium hyperplasia ,effusion and erosion between the two groups . The number of tophi was correlated with estimated glomerular filtration rate ( eGFR) ,serum creatinine( SCr) and blood urea nitrogen ( BUN) ( rs = - 0 .595 ,0 .511 ,0 .583 ;all P < 0 .001) ,respectively .Conclusions GCKD is more likely to manifest multiple tophi ,multiple tophus-involved articulations ,intra-tendinous tophi and double contour sign .The number of tophi is correlated with the degree of renal impairment .

Objective To explore the effect of action research based on group learning in hospice care module for nursing students. Methods 82 students were divided into cooperative learning group (40) and conventional teaching group (42). The cognition and attitude of good death questionnaire was used to evaluate teaching effect before and after teaching of hospice care. After teaching, self-feedback and self-feedback questionnaire were made to evaluate the effect of cooperative learning group. SPSS 17 was used to make comparison of data between groups and within groups, and t test, chi square test or Fisher exact probability 2 were also used . Results There were no difference between two groups of nurses before teaching. The students' cognitive level and attitude in cooperative learning group increased significantly, especially their cognitive level of physiological needs (t=5.398, P=0.001), cognition and attitude of death education in intervention group was higher than control group (t=2.992, P=0.004; t=3.661, P=0.001). Although 95% of the nursing students of the cooperative learning group thought group cooperative studying could improve their interests and cultivated their comprehensive abilities, only 50% could accept this kind of studying methods . Conclusions Action research based on group cooperative studying can improve students' cognitive level and attitude of good death, and it is also good for cultivating their ability of au-tonomous learning, innovation, cooperation, and meanwhile, educational action research is a good way to improve both teaching effect and research ability of teacher staffs, but in practice, students' acceptance of group learning needs to be improved.

OBJECTIVEThis study aimed to investigate the effect of a lactoperoxidase-peroxidase-thiocyanate (LPO-H2O-SCN-) system with different concentrations of iodine (I-) on Streptococcus mutans (S. mutans), particularly on various parameters, including growth, adhesion, glucosyltransferase (GTF) enzyme activity, and insoluble exopolysaccharide synthesis.

METHODSS. mutans ATCC 25175 was used as experimental species. Clonal formation unit (CFU) were counted to investigate the inhibitory effect on bacterial growth. The inhibition rate of bacterial adherence was calculated to analyze the effect on adhesion. Anthrone method was used to determine the content of insoluble exopolysaccharides and the amount of reducing saccharides. GTF activity and enzyme activity were then determined.

RESULTSThe inhibitory ability of the LPO-H2O2-SCN- system with I- on the cariogenicinity of S. mutans was strengthened as I- concentration was increased. At I- concentration > or = 100 micromol x L(-1) the antibacterial effects were significantly increased compared with those of the control group (P < 0.05). At I- concentration > or = 1,000 micromol x L(-1), the antibacterial effects were significantly improved compared with those of the group with SCN-only (P < 0.05). At I- concentration > or = 100 micromol x L(-1), the inhibition rate of bacterial adherence was > 50%; insoluble exopolysaccharide synthesis and GTF enzyme activity were reduced (P < 0.05).

CONCLUSIONThe antibacterial effects of the LPO-H2O2-I- system were enhanced by adding I- to overcome the antagonistic effect of physiological SCN- concentration. LPO-H2O2-SCN- system with different concentrations of I- showed statistically significant inhibitory effects on growth, adhesion, insoluble exopolysaccharide synthesis, and GTF enzyme activity.

Objective:To investigate the diagnostic value of the promoter methylation of plasma RNF180 gene and its protein ex-pression for the detection of gastric cancer. Methods:Methylation-specific polymerase-chain reaction (MSP) and enzyme-linked immu-no-sorbent assay (ELISA) were performed to detect DNA methylation and protein expression of the RNF180 gene, respectively. The correlations of DNA methylation and protein expression of the RNF180 gene with the clinico-pathological parameters of gastric carcino-ma were then separately analyzed. Results:MSP showed that the methylation rates of the RNF180 gene were 62.75%and 21.88%in the plasma of patients with gastric carcinoma and healthy volunteers, respectively;this result indicated that the two groups significantly differed (P<0.01). The methylation of the RNF180 gene was associated with tumor size, clinical stage, tumor differentiation, lymph node metastasis, and distant metastasis (P<0.05). ELISA results showed that the protein expression of the RNF180 gene [(23.22 ± 1.36)μg/mL] was significantly lower (P<0.01) in the plasma of patients with gastric carcinoma than in the plasma of healthy volunteers [(34.25 ± 2.44)μg/mL]. However, the protein expression of the RNF180 gene was not associated with clinicopathological parameters (P>0.05). Conclusion:The RNF180 gene is expressed at a hypermethylation rate, and the corresponding protein expression level is de-creased in the plasma of individuals with gastric carcinoma. Therefore, RNF180 gene methylation in plasma could be applied to detect microinvasion for the clinical diagnosis of gastric cancer.