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.

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.

Abstract: Objective To establish a rapid detection assay based on fluorescence recombinase polymerase amplification (RPA) targeting Necator americanus eggs, and to evaluate its efficacy, providing technical support for rapid detection of Necator americanus in fecal samples. Methods The fluorescence RPA primers and probe were designed based on the cox1 gene of Necator americanus and then screened the optimal combination to develop the assay. The genomic DNA of Necator americanus eggs was diluted to 7 concentration gradients including 100 pg/µL, 10 pg/µL, 1 pg/µL, 100 fg/µL, 10 fg/µL, 1 fg/µL, 0.1 fg/µL, to determine the detection limit of the assay. The specificity of the assay was demonstrated by detected genomic DNA from Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis and Fasciola hepatica. A total of 44 fecal samples were collected and DNA extraction was performed, and the modified Kato-Katz method, semi-nest PCR method, and fluorescent RPA method were simultaneously used for detection to evaluate the sensitivity and specificity. Results The established fluorescence RPA assay can specifically amplify a fragment of 194 bp of the Necator americanus cox1 gene within 20 min, with a detection limit of 10 fg/µL. There was no cross-reactivity with Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis, Fasciola hepatica after specificity validation. In 44 fecal samples, 27 positive samples were detected by the fluorescence RPA assay, and 26 positive samples were detected by both the Kato-Katz and the semi-nested PCR. The fluorescence curve of sample number 1 was slightly higher than the negative control in the later stage of the reaction, but did not show a similar trend to the positive control, and was therefore judged to be a suspected negative sample. Compared with the Kato-Katz method and the semi-nest PCR method, The sensitivity of the fluorescent RPA method were 100.00% and the specificity were 94.44%, and the consistency of the detection results was good (Kappa=0.953>0.75). Conclusions The assay based on the fluorescence RPA is an efficient, sensitive and specific technique for detecting Necator americanus and it can be applied for surveillance and early warning of hookworm infection.

Abstract: Objective To establish a sensitive and specific nucleic acid detection method for Schistosoma japonicum based on loop-mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR) technology. Methods The LAMP primers, gRNA and ssDNA probe that target Schistosoma japonicum SjR2 genes were designed according to the principles of LAMP and CRISPR. The LAMP-CRISPR reaction system was established and optimized. The sensitivity and specificity of the method were evaluated against the ten-fold serial dilutions of plasmid containing SjR2 target sequences, as well as genomic DNA at different stages of Schistosoma japonicum and other parasites, including Fasciola hepatica, Schistosoma mansoni, Taenia saginata, Clonorchis sinensis, Ascaris lumbricoides, Necator americanus, Paragonimus westermani, and Echinococcus granulosus. Additionally, 15 schistosome-infected snail and 30 uninfected samples were tested by LAMP-CRISPR and LAMP methods, respectively, to evaluate the potential of this method for screening for infected snails. Results　The developed LAMP-CRISPR method was able to specifically amplify and detect the SjR2 gene of S. japonicum. The optimal reaction temperature was 37 ℃, and the optimal reaction concentrations were both 40 nmol/L for gRNA and Cas12a protein. No cross-reaction was observed with genomic DNA from other parasites such as F. hepatica. The detection limit of the method was 10 copies/μL when testing 10-fold dilutions of recombinant plasmids as a template. Furthermore, the LAMP-CRISPR method was able to accurately detect genomic DNA from S. japonicum at various stages of development, including eggs, cercariae, schistosomula, juvenile worms, and adult worms. The results of testing 45 snail samples showed no significant difference between the LAMP-CRISPR and LAMP methods for detecting infected snails (χ2=0.05, P>0.05). The sensitivity and specificity of the LAMP-CRISPR method were 100.00% (15/15) and 96.67% (29/30), respectively, compared to the gold standard, while the sensitivity and specificity of the LAMP method were 100.00% (15/15) and 93.33% (28/30), respectively. Conclusions　This established LAMP-CRISPR detection method presented good sensitivity, specificity and reliability, making it a promising tool for rapid detection and risk monitoring of S. japonicum.

Whether to select one-tailed test or two-tailed test,how to establish null hypothesis and alternative hypothesis,how to improve the test efficacy were elaborated,and the common problems encountered when hypothesis was applied in scientific papers were pointed out with examples.

In this study, the flow injection (FI) technology combined with the unique step wise multiple ions monitoring scanning (step-wise MIM) mode of Qtrap-MS was first established for the identification and discrimination of eight Murraya species. It only takes 5 min for each sample to detect approximate 600 compounds. The characteristic MS chromatograms of eight Murraya plants were analyzed by Analyst and SIMCA-P. The results of PCA showed that sect. Murraya and sect. Bergera were clearly divided into two categories, suggesting that there is difference in the chemical compositions between these two groups. Further detail analysis of the MS data could realize the preliminary structure elucidation of the component types contained in different plants. The main components in M. exotica and M. alata are coumarins, and polymethoxyflavones are rich in M. paniculata, while carbazole alkaloids are the major ones in sect. Bergera plants. The results are consistent with our previous comprehensive analysis of the chemical components of Murraya species. In conclusion, our research confirmed that FI-Qtrap-MS technology can be used for rapid identification and differentiation of similar plant species, providing reference for chemical taxonomy and a new method for the quality evaluation of medicinal materials.

This study was aimed to explore the antitumor effect of volatile oil of X ihuang pill and its immune mecha-nism in order to screen the antitumor active site of Xihuang pill. Among 70 female Wistar rats, 10 rats were random-ly selected as the blank control group; and the other 60 rats was used in the establishment of walker 256 breast can-cer cell tumor-bearing rat models. The model rats were randomly divided into the negative control group (model group), high-dose volatile oil group, middle-dose volatile oil group, low-dose volatile oil group, high-dose Xihuang pill group, and lentinan group (positive control group), with 10 rats in each group. The intragastric administration was given twice a day for 14 days. Blood was taken from the abdominal aorta. Tumor tissues was removed and weighed to calculate the tumor inhibitory rate. ELISA method was used to detect the level of IL-2, IL-6, IL-10, IFN-γ and TGF-β in peripheral blood. The flow cytometry was used to detect the content of CD3+ T cell, CD4+ T cell, CD8+ T cell, and B7-1 cell (CD80). The results showed that the tumor inhibitory rates of volatile oil of high-dose group and middle-dose group were 28.4% and 24.1%, respectively. Compared with the model group, the average level of IL-2 and IFN-γ of volatile oil of high-dose group and middle-dose group and CD3+ T cell, CD8+ T cell, B7-1 cell con-tent were obviously increased (P< 0.05). It was concluded that volatile oil of Xihuang pill had certain antitumor ef-fect, which was one of the antitumor active sites of Xihuang pill. The volatile oil of Xihuang pill upregulates the lev-els of IL-2 and IFN-γ, as well as the contents of CD3+ T cell, CD8+ T cell, B7-1 cell in order to increase the im-mune clearance function of tumor-bearing rats.

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.