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 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.

In proteomic research, to improve protein solubility of membrane proteins and nuclear proteins, buffers containing high concentration of detergent, such as 4% SDS, were widely used. However, high concentration of detergent might severely interfere with the downstream proteomic analysis, including protein quantitation and trypsin digestion. To improve the proteomic compatibility of buffers with high concentration of detergent, we used short gel method to pretreat buffers containing detergent. Protein samples were first separated by a short (2-2.5 mm) SDS-PAGE electrophoresis, and proteins were quantitated by comparing with bovine serum albumin standards via optical density analysis. The gel was then cut and peptides were recovered using in-gel digestion. The quantitative linearity range of this method was 1 to 8 μg. The quantitation was accurate and reproducible. After short gel analysis, recovered peptides generated high mass spectrometry signals. In conclusion, short gel method eliminated the interference of high concentration detergent in the proteomics analysis, and it was suitable for protein samples' pretreatment, and was worth to apply in proteomic research.
Detergents ; chemistry ; Electrophoresis, Polyacrylamide Gel ; methods ; Mass Spectrometry ; Membrane Proteins ; chemistry ; Nuclear Proteins ; chemistry ; Proteins ; chemistry ; Proteomics ; methods ; Trypsin

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.

PurposeTo explore the value of prenatal ultrasound in diagnosis of fetuses with limb body wall complex (LBWC).Material and MethodsThe ultrasound images and follow-up results were studied retrospectively in 20 cases of LBWC, and the ultrasonic features of LBWC were analyzed.ResultsAll 20 cases showed rather severe body wall defect and spinal abnormality, including 16 cases of abdominal wall defect, 4 cases of abdominal thoracoabdominal defect, 8 cases of idiopathic scoliosis, 2 cases of kyphosis, 3 cases of twist into a corner, 1 cases of distortion as S shape, and 6 cases of vertebral body arranged in disorder. Moreover, 16 cases presented abnormal umbilical cord,among which 11 had too short umbilical cord, 1 without umbilical cord, and 7 as single umbilical artery (3 cases with short cord). Five cases showed limb abnormalities, among which 2 cases were left lower limb absence, 1 cases as left upper limb absent, 1 cases as left foot varus and right foot slightly varus, and 1 cases as left foot drop.ConclusionPrenatal ultrasonography can accurately diagnose LBWC in time, so as to provide evidence for early clinical intervention.