Objective To observe the expression of tumor stem cell markers P 75NTR,Oct-4,Sox-2,Lin28 and Nanog in the tumor sphere from esophageal squamous cells carcinoma Eca 109 and identify the esophageal squamous cell cancer stem cell marker .Methods The serum-free culture method was used for generating tumor spheres: proliferation was observed in enrichment culture tumor spheres .Small tumor spheres were obtained after 5 days culture and big and round tumor spheres appeared after 14 days culture which were collected for experiments and passaged .The expression and location of P75NTR,Oct-4,Sox-2,Lin28, and Nanog were detected by immunofluorescence cytochemistry .Results The expressions of P75NTR,Oct-4 and Lin28 were positive in the center of tumor spheres and some on cytoplasm and other in nuclei of Eca109 monolayer cells.However, Oct-4 fluorescence intensity was weaker than P75NTR.The expressions of Sox-2 and Nanog were positive in cytoplasm of tumor spheres and Eca 109 monolayer cells .Conclusion The cells expressing P75NTR, Oct-4, and Lin28 in the center of the tumor sphere may be esophageal cancer stem cells .

OBJECTIVE：To provide refere nce f or the quality control and evaluation of rice-wine processed Coptidis Rhizoma decoction piece. METHODS ：Taking 17 batches of rice-wine processed Coptidis Rhizoma decoction piece from different manufacturers as samples ，HPLC method was adopted to determine the contents of 4 kinds of alkaloids as epiberberine ，coptisine， palmatine and berberine. The compound weights of epiberberine ，berberine，palmatine and berberine were calculated by the subjective and objective combination weighting method （AHP combined with variation coefficient ）. Then the quality evaluation method was used to evaluate the quality of decoction pieces combined with the appearance of decoction pieces and the contents of 4 alkaloids. The percent mass constant was calculated and the grade of rice-wine processed Coptidis Rhizoma decoction piece was classified. RESULTS ：According to the results of content determination of 4 kinds of alkaloids ，among 17 batches of samples ，a total of 13 batches of samples met the requirements of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. Mass constants of 13 batches of qualified samples were 10.03-26.96，and the percent mass constants were 37.20％-100％. If the percent mass constant ≥ 80％ of the sample was listed as the first-class product ，the sample with the percent mass constant between 50％-＜80％ was classified as the second-class product ，and the rest was listed as the third-class product ，therefore the quality constant of first-class product was ≥21.57，that of second-class product was 13.48-＜21.57，and that of third-class product was ＜13.48. According to the grading standard ，3 batches of 13 batches of qualified samples are classified as first-class products ，6 batches are classified as second-class products ，4 batches are classified as third-class products. CONCLUSIONS ：The established subjective and objective combination weighting method and quality constant method can more scientifically and reasonably classify rice-wine processed Coptidis Rhizoma decoction piece.

Objective To investigate the effect from circumferential distribution angle of forwards wedge vortex generators on aerodynamic drag reduction during flow around a cylinder, so as to provide theoretical evidences for low drag sprint garment design. Methods Forwards wedge vortex generators were reconstructed based on the NIKE’s AeroBalde. Given that the individual parts of an athlete body can be treated as multiple cylinders with varied dimensions and positions, 48 forwards wedge vortex generators were distributed as four columns on windward side of the cylinder, which were symmetrical with the YOZ plane. When the air flowed through the cylinder at the speed of 32 km/h, large eddy simulation was carried out on the computational domains which were properly meshed with polyhedral mesh to investigate the drag force, flow filed and pressure distributions. Results It was effective for drag reduction of the cylinder induced by airflow when two columns of forwards wedge vortex generators were circumferential distributed in the range of 55°-75°and the circumferential internal angle between two columns was in the range of 10° or 15°. The pressure distribution on leeward side of the cylinder was apparently changed after surface modification, which minimized the pressure drag dominant in aerodynamic drag. The drag reduction mechanism was that micro-vortices were generated downstream after flow through the forwards wedge vortex generators, which resulted in an early transition to critical flow with low drag force. Conclusions Effective aerodynamic drag reduction is achieved if forwards wedge vortex generators are properly distributed. The research findings can provide guidance for wind tunnel test and low drag sprint garment design.

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.