Objective:To clone soluble human anti-digoxin antibodies from large phage antibody library and construct a vector that expresses diabody.Methods:Soluble ScFvs were prepared through infecting E coli.HB2151 with the selected phage antibodies and induced with IPTG.The diabody vector was modified by enzyme digestion and checked by SDS-electrophoretogram.Results:It was showed by ELISA that soluble ScFv had specific binding ability to digoxin.The vector to express a soluble diabody was obtained by genetic modification,which was shown by Western blot.Conclusion:Soluble human anti-digoxin antibodies were successfully obtained from phage antibodies.The vector is efficient in creating diabody.

Objective To investigate the application value of DFY-Ⅱ ultrasound imaging analysis software in evaluation of fetal lung maturity in different pregnancy.Methods Totally 315 cases of healthy single pregnancies (24-41 weeks) were collected.The sonographic views of fetal right side sagittal plane were obtained.The echo intensity of fetal lung and liver were analyzed and the ratio was canculated with DFY-Ⅱ ultrasound imaging analysis software.Results The ratio of echo intensity from fetal lung and liver had positive correlation with gestational weeks (r=0.94,P＜0.05),the linear regression equation was Y=0.60+0.07X (r2 =0.883).Conclusion The ratio of echo intensity by fetal lung and liver analyzed with DFY-Ⅱ ultrasound imaging analysis software can be an effective method to evaluate fetal lung maturity.

Objective To investigate whether Tumor Necrosis Factor-alpha (TNFα) is capable of activating Rho kinase pathway which leads to smooth muscle cell proliferation and the intervention function of Rosuvastatin, and clarify the mechanism and intervention manner of anti-atherosclerosis by Rosuvastatin. Methods Wistar neonate rat smooth muscle cells were cultured, and the activity of cell proliferation was determined by methyl thiazolyl tetrazolium (MTT). The expression of Rho kinase genes after the stimulation of TNFα was evaluated by RT-PCR. Western blot method was used to measure the protein expression of proliferating cell nuclear antigen (PCNA) after TNFα stimulation and Rosuvastatin intervention in smooth muscle cell. Results The TNFα stimulation significantly enhanced the expression of Rho kinase and increased the expression of PCNA protein in smooth muscle cells (P < 0.05). These effects were positively correlated with prolonged treatment whereas additional Rosuvastatin administration inhibited the above-mentioned effects (P < 0.05). Conclusions The activation of TNFα mediated Rho kinase signaling pathway can significantly promote smooth muscle cell proliferation, and Rosuvastatin can not only inhibit this pathway but also the induced proliferation.

Objective To determine the expression of indole-2,3 dioxygenase (IDO) in human acute leukemia,and to investigate its correlations with clinicopathological parameters and prognosis in acute leukemia.Methods The expression of IDO in protein and RNA levels was detected by immunohistochemistry and real-time quantitative RT-PCR,respectively,and the correlations of IDO with clinicopathologic features and prognosis of acute myeloid leukemia (AML)-M5 were analyzed.Results The positive rate of IDO protein was 63.3 ％ (38/60) in human acute leukemia,while it in AML (34/49,69.4 ％),especially in AML-M5 patients (29/35,82.9 ％),was significantly higher than that of acute lymphoblastic leukemia (4/11,36.4 ％).The expression of IDO protein in healthy human peripheral blood mononuclear cells was negative.The RNA expression level of IDO in AML-M5 or non AML-M5 patients were significantly higher than that of healthy people (P ＜ 0.001),and AML-M5 patients had significantly higher IDO RNA level than that in non AML-M5 patients (P ＜ 0.05).The IDO gene expression was not correlated with sex,age and drug sensitivity,while it was closely related with these factors in the patients without complication of pulmonary infection.IDO could not act as an independent prognostic marker.Conclusion The expression of IDO in AML-M5 patients is significantly higher than that in non AML-M5 patients and healthy people.The positive expression of IDO is associated with poor prognosis of AML-M5 patients,but it is not an independent poor prognostic indicator.

Periampullary carcinoma is a rare malignant tumor of digestive system,and its accurate diagnosis is still difficult.From January 2007 to July 2012,12 patients with periampullary carcinoma had been admitted to the Southwest Hospital of Third Military Medical University,and the imaging data were retrospectively analyzed.The results of ultrasonography revealed that all tumors were hypoechoic.The tumor displayed hyperenhancement in 3 patients,isoenhancement in 1 patient,hypoenhancement in 8 patients during the arterial phase on contrastenhanced ultrasonography (CEUS),while the tumor displayed hypoenhancement in all patients during the venous phase.Magnetic resonance imaging (MRI) plain scanning showed duodenal papilla enlargement in 1 patient,ampullary tissue mass signal in 2 patients,tissue mass signal at distal common bile duct in 2 patients,the rest 7 patients did not show tissue mass signal.Lower biliary obstruction was the common manifestation of the 12 patients on magnetic resonance cholangiopancreatography (MRCP),intrahepatic and extrahepatic bile vine-like expansion in 4 patients,double duct sign in 2 patients,the bottom of common bile duct with filling defect in 2 patients and it revealed beak-like narrow in 1 patient.CEUS,MRI and MRCP could both play an important role as conventional methods in diagnosing periampullary carcinoma.

Objective To Study the effects of high mechanical index ultrasound radiation microbubble on colon cancer cell skeletal.Methods Lovo cell were cultured in vitro,and this study was divided into control group,simple contrast agent group,microbubble plus ultrasound radiation group and simple ultrasound radiation group.Ultrasound contrast agent SonoVue and ultransound (frequency 1.5 MHz and MI 1.7) were used.Results In the control group,the microtubes were densest in dyeing silk,and extended to the edge of a cell.In the ultrasound and microbubble group,the microtubes were weaker and thin,the.network was mainly in the structure of the long axis.The cells arranged in a slight pigmentation to the density cell,to reach out many fine short hair,there were obviously a sense of silk and directional.In the microbubble and ultrasound group,Lovo cells streamed of the central significantly reduced,fluorescent lighting,with short hair.There was no difference in the ultrasound group and the microbubble group with microfilament and microtubule.Conclusions High mechanical index ultrasound radiation can changed into vacant,a trace of the assembly and distributed to the tumor cells are attacking,the transfer of restraining.

Objective To evaluate intraoporative ultrasonography (IOUS) of liver tumors. Methods In this study, 45 patients with liver tumors were examined by the IOUS(Logiq 500 scanner with a dedicated 5 ～ 7.5 MHz linear-array intraoperative transducer), and the results of IOUS were compared with those of preoperative ultrasonography (PHILIPS HDI 5000 scanner with a 3.5 ～ 5 MHz convex-array transducer) and computed tomography (CT). Results For tumors with a diameter ≥ 1 cm, the detection rate of IOUS was 100%, which was higher than that of preoperative ultrasonography and CT (100% vs 89.80% and 97.96%), though there was no statistical significance(xc2 = 3.372 ,P >0.05 and Fisher exact probability was 1.00) ; For tumors with a diameter < 1 cm, IOUS had a significantly higher detection rate than preoperative uhrasonography and CT(90.70% vs 62.79% and 74.42%, respectively,the former x2= 9.382, the latter x2 = 3.957, both P < 0.05) ;The total detection rate of IOUS was significantly higher than that of preoperative ultrasonography and CT (95.65% vs 77.17% and 86. 96%, respectively, the former x2 = 13.378, the latter x2 = 4.381, both P < 0.05). All the detected lesions underwent surgical resection or intratumoral injection of ethanol with the guidance of IOUS. Conclusions Compared with preoperative imaging examination, IOUS not only can enhance the detection rates of preoperatively overlooked lesions, but also can provide more valuable information to guide the surgical procedures.

Objective To observe the perfusion pattern of hepatic angiomyolipomas using contrastenhanced ultrasound(CEUS) and compare diagnostic efficacy of contrast-enhanced ultrasound with contrastenhanced helical CT(CECT).Methods Nineteen patients with 21 resected and pathologically proven hepatic angiomyolipomas were included in this study.Low mechanical index (mechanical index less than 0.2) realtime CEUS was performed in nineteen patients (5 patients with pulse inversion harmonic, 14 patients with contrast pulse sequencing, CPS) after 2.4ml bolus injection of contrast agent SonoVue.CECT was performed using 16-slice helical CT and contrast agent Ultravist.The diagnostic performance was calculated by considering histologic results as the reference standards.Results Fifteen tumors were correctly diagnosed as hepatic angiomyolipomas,4 tumors were misdiagnosed (including 1 hepatocellular carcinoma,2hepatic adenomas, 1 hemangioma) and 2 tumors were characterized as benign lesions by CEUS.Eight tumors were correctly diagnosed as hepatic angiomyolipomas, 13 tumors were misdiagnosed (including 7hepatocellular carcinoma, 1 liposarcoma, 2 lipomas, 1 hepatic adenomas, 2 hemangiomas) by CECT.The preoperative diagnostic accuracy was 71.4% for CEUS and 38.1 % for CECT(P＜0.05 ).Conclusions CEUS can demonstrate typical imaging characteristics of most hepatic angiomyolipomas, and has higher diagnostic performance than CECT in characterization of hepatic angiomyotipomas.