Objective To study the effects of As_2O_3 on tumor model of hepatocarcinoma.Methods HepAgrafed hepatocarcinoma mouse model was established by subcutaneously injection of mouse hepatoma cells(1×10~6)into the oxter of mice.After treated by As_2O_3,the volume change of tumor and tumor inhibition rates were observed.The expression of vascular endothelial growth factor(VEGF) was detected by immunohistochemical and calculated the difference of MVD.Results The volume of tumor and the tumor inhibition rates were significantly decreased in As_2O_3 group compared with control group(P<0.05).The As_2O_3 could inhibit angiogenesis of xenograft tumor,depress expression of VEGF and decrease microvascular density(MVD).Conclusion As_2O_3 can inhibit the growth of tumor,inhibit the expression of VEGF and decrease MVD.

To understand the immunogenicity of purified inactivated severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), concentration by ultrafiltration as well as molecular-sieve chromatography (MSC) were used for purification of inactivated SFTSVs. Inactivated viruses in purified samples were analyzed and identified by western blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the glycoprotein (GP) and nucleoprotein (NP) antigen titers of which were detected using a double-sandwich enzyme-linked immunosorbent assay (ELISA). Purified inactivated SFTSVs were enriched and observed by electron microscopy, and the total protein concentration detected using the bicinchoninic acid assay. Purified inactivated SFTSVs were applied to New Zealand rabbits via two immunization programs to evaluate immunogenicity and to compare the immune effect. After SFTSVs were inactivated and concentrated by ultrafiltration, MSC revealed two typical elution peaks. The sample of one peak was identified as inactivated virions, in which GP and NP were detected by SDS-PAGE, western blotting and ELISA. Main corponent of the other peak was NP. After concentration by ultrafiltration, purified inactivated SFTSVs with purity >90% and total protein concentration of 1. 1 mg/mL were obtained, and the typical electron microscopy of bunyavirus was observed. In the sera of animals immunized with purified inactivated SFTSVs, SFTSV-specific IgG antibody and neutralizing antibody were detected at high titers. However, antibody titers were affected by the immunization program. Effect of immunization on days 0, 14 and 28 was significantly better than that on days 0, 7 and 28. Our work revealed that cultivation of SFTSVs contained intact virus particles and large amounts of free NP. Using MSC, purified inactivated SFTSVs of high purity could be obtained. Purified inactivated SFTSVs induced high titers of neutralizing antibody and virus-specific IgG antibody showing satisfactory immunogenicity, which provides important clues for further study on a vaccine for the inactivated virus.
Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Bunyaviridae Infections ; immunology ; virology ; Humans ; Neutralization Tests ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

To prepare monoclonal antibodies (mAbs) against structural proteins of severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), BALB/c mice were immunized using purified inactivated SFTSV virions as the antigens. Subsequently, hybridoma cell lines that secreted monoclonal antibodies against nucleoprotein (NP) and glycoproteins (GP) were obtained using a hybridoma technique. The antigen specificities of prepared mAbs were examined by indirect immunofluorescence and immunoprecipitation assays. Functional analyses were then performed,including the detection of IFA antibody titers,the levels of neutralizing activity and antibody affinities. After cell fusion and cloning,13 hybridoma cell lines secreted mAbs specifically against SFTSV-GP and 7 hybridoma cell lines secreted mAbs specifically against SFTSV-NP. Immunofluorescence and immunoprecipitation assays showed that the mAbs had high levels of antigen specificity. Among the 13 anti-SFTSV-GP mAbs,6 recognized Gn,whereas the others reacted with Gc. IFA titers of most anti-SFTSV-GP mAbs were between 1,280 and 20,480, and four anti-SFTSV-Gn mAbs showed neutralizing activity. Seven of the obtained anti-SFTSV-NP mAbs reacted specifically with NP,of which the IFA titers ranged from 5,120 to 20,480 with no observed neutralizing activity. Furthermore, two anti-SFTSV-GP mAbs, 1C8 and 1G8, showed high levels of affinity via a non-competitive ELISA. Our study lays the foundation for the development of further diagnostic assays and basic research into SFTSV.
Animals ; Antibodies, Monoclonal ; immunology ; Antibodies, Viral ; immunology ; Antibody Specificity ; Bunyaviridae Infections ; immunology ; virology ; Female ; Humans ; Hybridomas ; immunology ; Mice ; Mice, Inbred BALB C ; Phlebovirus ; immunology ; Viral Structural Proteins ; immunology

This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

Acetates ; therapeutic use ; Acute Disease ; Anti-Asthmatic Agents ; therapeutic use ; Asthma ; prevention & control ; Child ; Child, Preschool ; Female ; Humans ; Leukotriene Antagonists ; therapeutic use ; Male ; Quinolines ; therapeutic use

Three pSIREN-siRNA plasmids were constructed using a pSIREN-RetroQ vector to silence the expression of muhidrug resistance-associated protein (MRP1) gene, and subsequently characterized by restriction endonuclease digestion and DNA sequencing. A truncated MRP1 and a full-length MRP1 were cloned into pEGFP-N2 and PeDNA3.1 respectively as pEGFP-MRP1T and pcDNA-MRP1. The plasmid pEGFP-MRP1T was co-transferred with each of the three pSIREN-siRNAs into HEK293 cells for MRP1T-GFP targeted silencing, and pSIREN-siRNA1 was used as the negative control, pSIREN-siRNA2 and pSIREN-siRNA3 appeared to be more effective to silence MRP1T-GFP compared to pSIREN-siRNA1 as shown by fluorescence microscopy. For the silencing of full-length MRP1 expression, HEK293 ceils were co-transferred with pcDNA-MRP1 and either of the three pSIREN-siRNAs, then subjected for Western blot analysis and MTT assays, pSIREN-siRNA2 and pSIREN-siRNA3 were able to inhibit the expression of 190 kD MRP1, but not pSIREN-siRNA1. The MDR of MRP1-transfected HEK293 ceils was abolished with pSIREN-siRNA2 or pSIREN-siRNA3 transfections. RNA secondary structure predictions demonstrated that the mRNA local free energy (△G) of the siRNA1 targeted sequence was lower, as the GC content and Tm value of siRNA1 were higher than those of siRNA2 and siRNA3. These data suggest that the local structure siRNAs and target mRNA may influence the silencing efficiency of MRP1 expression.

Activating protein-2 (AP-2) is a cell type-specific DNA binding transcription factor family with the ability to regulate the expression of specific target genes. Five isoforms of AP-2 have been discovered, they are AP-2alpha, AP-2beta, AP-2gamma, AP-2delta and AP-2epsilon. AP-2s are involved in the regulation of cell proliferation, differentiation and apoptosis as well as embryogenesis of mammary animals. Recently, the function of AP-2 in neoplasm has attracted increasing attention. Researches reveal that the modulation of AP-2 in tumorigenesis may be dual, either inhibitory or promoting, which depends on the specific tissues,stages of cancer progression and difference between five family members. This review summarizes recent research progress on the role of AP-2 in the oncogenesis and their potential applications in clinical practice.
Animals ; Cell Transformation, Neoplastic ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasms ; etiology ; genetics ; Transcription Factor AP-2 ; genetics

ObjectiveTo discuss the application value of high frequency ultrasound (HFUS) characteristics, acoustic radiation force impulse (ARFI) imaging technology and contrast-enhanced ultrasound (CEUS) in the diagnosis of soft tissue hemangioma.MethodsTo retrospectively analyze the clinical data of 44 cases of soft tissue hemangioma that were treated in Capital Medical University Affiliate Beijing Chaoyang Hospital from August, 2013 to May, 2014, and to analyze the difference between the characteristics of HFUS, ARFI and CEUS in soft tissue hemangioma and normal surrounding tissue.ResultsHFUS shows the features of morphological diversity of sinus shape expansion tube structure, unclear boundary, irregular configuration, compressibility and partial strong echo in the phlebolith. Color Doppler ultrasound (CDFI), detects abundant interphase red-blue bloodstream signal slowly and consistently. The blood signal is strengthened after partial compression. CDFI shows more vein spectrum in the lesion. The discrepancy of comparison between VTQ and SWV value of soft tissue hemangioma and surrounding muscular tissue possesses statistical significance [(1.082±0.183) m/svs (1.414±0.331) m/s,P<0.01]. Ultrasound contrast can show the relationship between diseased region and surrounding tissue clearly, which is beneficial to the selection of operation method and prognosis.ConclusionThrough conventional two-dimensional ultrasound and high frequency CDFI, and further combining the acoustic pulse radiation ARFI technology and CEUS technology, the soft tissue hemangioma can’t only be more accurately diagnosed, but also provides more reliable diagnostic basis for clinic.

ObjectiveTo investigate the features of normal and psoriasis skin on ultrasound biomicroscopy (UBM) and explore the method of thickness measurement.MethodsUsing 50 MHz ultrasound probe of biological microscope, ultrasonographic observation and ultrasonic thickness measurement were conducted in 90 normal adults and 40 psoriasis patients. Innormal patients, ultrasound evaluations were performed at 10 different parts of the body skin.ResultsOn sonogram, the normal skin showed a “sandwich” structure with two parallel hyperechoic bands and the middle isoechoic dots and short liens. The sonograms of the psoriasis skin showed obviously thickened epidermis and dermis, disordered internal structure and clear boundary from adjacent normal skin. The range of the epidermis’ thickness measurement was between the medial forearm (0.12±0.03) mm and the palm (0.29±0.15) mm. The range of dermal thickness measurement was between the back hand (1.18±0.32) mm and parasternal (1.55±0.21) mm. Psoriasis skin was thicker than the uninvolved skin (P<0.001). And the dermis’ thickness of uninvolved skin in psoriasis patients was thicker than that of the normal adults (P<0.001).ConclusionNormal adult’s epidermis, dermis and skin appendages can be shown clearly using 50 MHz ultrasound biomicroscopy. And ultrasound biomicroscopy canaccurately measure the thickness of dermis and epidermis, which provides the basis for the diagnosis and treatment of psoriasis.

Objective To evaluate the value of virtual touch quantization (VTQ) imaging in diagnosis of IgA nephropathy. Methods The clinical data of 85 patients with IgA nephropathy were analyzed, who were treated in Capital Medical University Affiliate Beijing Chaoyang Hospital from December 2013 to July 2014. The patients who was with critical condition, unable to cooperate and with other pathological types were excluded. Finally 108 kidneys of IgA nephropathy with mesangial cell hyperplasia in 54 cases were included into the study. Meanwhile 108 kidneys in 54 volunteers who took the health physical examination in our hospital were taken as healthy controls. VTQ was performed in middle part of kidney and the measurements of shear wave velocity (SWV) was recorded. The mean SWV of renal parenchyma and collecting system was compared in different groups. Results The mean SWV measurement of renal parenchyma and collecting system in control group were (2.13±0.13) m/s, (1.15±0.02) m/s;the results in IgA nephropathy group were (3.07±0.62) m/s, (1.12±0.29) m/s. The mean SWV of renal collecting system was lower than that of renal parenchyma (t=-14.481, P<0.001). The mean SWV of renal parenchyma and collecting system in IgA Nephropathy group was higher than that in control group (t=-54.01, P<0.001). The renal parenchyma VTQ value positively correlated with the degree of renal insufficiency for patients with chronic kidney disease (CKD) (F=798.70, P<0.001). The interlobular arterial resistance index (RI) increased gradually with CKD stage, but no statistical differences were found. Conclusion In terms of early diagnosis and clinical staging, VTQ technology has some diagnostic value in evaluation of renal parenchymal damage for patients with IgA nephropathy.