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.

AIM: To measure microcirculation function in type 2 diabetic patients and non-diabetic subjects with a new measurement method called capillary recruitment. METHODS: 276 type 2 diabetic patients in Shanghai downtown were enrolled and categorized into several groups, those with diabetes duration

Objective To construct the pshuttle-Egr-1-hSmac plasmid and transfect human breast cancer MDA-MB-435 cells,and to observe its radiotherapy enhancing effect on tumor cells.Methods The empty vector pshuttle and pshuttle-Egr-1-hSmac plasmid were transfected into MDA-MB-435 cells by liposomal.At different time(4,8,12,24 and 48 h)after irradiation with 2.0 Gy X-ray and 24 h after irradiation with 0.5 -5.0 Gy,the total RNA and protein were collected and extracted from these cells to analyze the Smac mRNA and protein expression levels with RT-PCR and Western blotting methods. The cells were divided into control, pshuttle, pshuttle-Egr-1-hSmac,2.0 Gy irradiation group, pshuttle + 2.0 Gy irradiation and pshuttle-Egr-1-hSmac+2.0 Gy irradiation groups.MTT method was used to evaluate cell proliferation,and the cell survival ability was measured with clone formation assay;Annexin Ⅴ/PI double staining and PI single staining were used to examine the apoptosis and cell cycle of MDA-MB-435 cells. Results There was no Smac mRNA expression in MDA-MB-435 cells in control and pshuttle groups,but the Smac mRNA expression levels in MDA-MB-435 cells in pshuttle-Egr-1-hSmac plasmid group were gradually increased with the time prolongation, and reached the maximum at 24 and 48 h;the Smac mRNA expression levels in MDA-MB-435 cells were increased gradually 24 h after irradiation of 0.5 - 5.0 Gy X-ray with the increasing of irradiation doses, and reached the maximum after 2.0 and 5.0 Gy irradiation. The Smac protein expression levels in pshuttle-Egr-1-hSmac plasmid group were increased gradually with the time prolongation,and reached the maximum at 24 h.The Smac protein expression lervels were increased 24 h afer irradiation of 0,0.5,1.0,2.0 and 5.0 Gy X-ray,especially in 5.0 Gy group. The MTT results showed that the A490 values in 2.0 Gy,pshuttle+2.0 Gy and pshuttle-Egr-1-hSmac groups 24, 48,and 72 h after irradiation were lower than those in control group(P<0.01);the A490 values of MDA-MB-435 cells in pshuttle-Egr-1-hSmac group after 1.0-5.0 Gy X-ray irradiation were lower than those in 0 Gy group (P<0.05 or P<0.01);the survival fraction(SF)in pshuttle-Egr-1-hSmac group was lower than those in control group (P<0.01).The percentages of the cells at G0/G1 and S phase in pshuttle-Egr-1-hSmac group were lower than those in 2.0 Gy group(P<0.01),the percentage of the cells at G2/M phase was higher than that in 2.0 Gy group (P<0.01);the apoptotic rate of the cells in pshuttle-Egr-1-hSmac group was higher than that in 2.0 Gy group (P<0.01).Conclusion X-ray irradiation can significantly increase the Smac mRNA and protein expression levels in MDA-MB-435 cells transfected with pshuttle-Egr-1-hSmac plasmid,inhibit the cell survival rate,and induce G2/M arrest and apoptotic increasing;Smac gene combined with radiotherapy could significantly increase the radiosensitivity of breast cancer cells.

Objective To optimize refinement of water extract from Bushen Yangxue Granules by chitosan flocculation.Methods According to the content of icariin detected by HPLC, the waters amount, extraction time and extraction times were evaluated by orthogonal design. The effects of the solution concentration, clarifying temperature and the amount of clarifying agent on the flocculation clarification processes were optimized with the content of icariin and polysaccharides.Results The optimum water extraction processes A2B1C3 were follows: 10 times amount of water, three times extraction and 1 h for each extraction process. The optimized flocculation clarification processes A1B2C3 were as follows: solution concentration was 0.4 g/mL, the clarifying temperature was 40℃ and the addition of chitosan was 0.1%.Conclusion The optimized refining process is stable and feasible.

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.

Objective To investigate the role of UDP-glucuronosyltransferase 1A (UGT1A),nuclear factor erythroid-2-related factor 2 (Nrf2) and kelch-like ECH-associated protein 1 ( Keapl ) in the tumorigenesis of colonic carcinoma.Methods The expressions of UGT1 A,Nrf2 and Keapl were detected in normal colonic mucosa(24 cases),adenoma tissue (30 cases) and adenocarcinoma tissue (77 cases) by immunohistochemistry,and the relationship between their expressions and the clinical pathological characteristics was analyzed.Results The positive rates of UGT1 A in normal colonic mucosa,adenoma and adenocarcinoma tissue were 83.3％ ( 20/24),80.0％ ( 24/30 ) and 53.2％ ( 41/77 ),respectively.The positive rate of UGT1A in adenocarcinoma was lower than those in colonic mucosa and adenoma ( all P ＜0.05 ).On the contrary,the positive rates of Nrf2 in adenoma [70.0％ (21/30) ] and adenocarcinoma tissue [ 87.0％ (67/77) ] were higher than that in normal colonic tissue [ 41.7％ (10/24),all P =0.000 ].The positive rates of Keapl in normal colonic mucosa,adenoma and adenocarcinoma tissue were 54.2％ ( 13/24),70.0％ (21/30) and 61.0％ (47/77),respectively ( normal colonic tissue vs adenocarcinoma tissue,P =0.040 ; adenoma vs adenocarcinoma,P =0.002 ).There was no correlation between the expression of UGT1 A,Nrf2 and the clinicopathologic features of colon carcinoma,while the differences of Keapl positive rates in the various degrees of tumor differentiation [ moderately-well differentiated vs poorly differentiated:70.0％ (35/50) vs 44.4％ (12/27) ] and invasion [T1-T2 vs T3-T4:78.8％ (26/33) vs 47.7％ (21/44) ]were statistically significant (all P ＜ 0.05 ).Conclusion The decreased expression of UGT1A and the dysregulation of Nrf2/Keapl system may play a role in colonic tumorigenesis.

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.

Objective To isolate HIV-specific T cell clone and to expand them in vitro through the activation-induced expression of CD137 molecule.Methods The peripheral blood mononuclear cells were isolated from HIV-infected patients and HIV Gag specific CD3+CD8+CD137+T cell subset were sorted to 96-well plate in 1 cell/well by multicolor flowcytometry and single cell sorting.After 14 days in vitro culture with feeder cells and cytokines, the numbers and phenotypes of the cultured HIV-specific T cells were calcu-lated and identified.Results The CD137 expression was low on rested T cells but up regulated by the stim-ulation with Gag peptide pool.The CD8+CD137+T cells could secret IFN-γ.The number of CD8 T cells reached to 106 after 14 days in culture and expanded to 107-108 cells after 28 days of culture in vitro 100%of the cells remained activated upon Gag stimulation.Conclusion In stead of using IFN-γ, CD137 could be utilized as a novel molecule to isolate and expand HIV specific T cells in vitro.The expanded antigen spe-cific T cell clones could maintain good activation status.