Preparation of Immuno-magnetic Beads and Their Separation & Detection to Ovary Cancer Cells

The organic monomer-molecule with nanometer magnetic powder by means of reforming the surface of nanometer magnetic powder have been synthesized. Magnetic beads in diameter of 2 mum or so am obtained by controlling conditions. Ovary cancer cells of ascites are separated and ovary cancer cells of blood arc detected by using immuno-magnetic beads linked with ovary cancer cell mono-antibodies. Results show that the specificity is 85% sensitivity is 87%, accuracy is 84%, cells acquiring purity is 90%, cells activity is 92% and detection sensitivity is 25 x 10(-7).

Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells

A simple and non-invasive method for detecting endometrial cancer in women with abnormal uterine bleeding is required. For this purpose, we prepared immuno-magnetic beads conjugated with anti-human EpCAM rat monoclonal antibody (mAb) for isolating exfoliated endometrial cells including endometrial cancer cells in vaginal discharge. The affinities of the anti-human EpCAM rat mAbs were analyzed by flow cytometry and immunocytochemistry and then magnetic beads were conjugated with the mAbs. The rate of retrieval of endometrial cells using the immuno-magnetic beads was calculated. Endometrial cells were isolated using the immuno-magnetic beads from the vaginal discharges of 22 patients with endometrial cancer and 16 non-malignant controls. The isolated cells were stained using endometrial cancer specific-mAbs and analyzed by flow cytometry and imaging cytometry. The immuno-magnetic beads conjugated with high-affinity mAb (clone 1456) appeared to have very low auto-fluorescence. Sufficient enrichment of Ep-CAMpositive cells using immuno-magnetic beads was observed in both simulation and clinical samples. The overall sensitivities of flow cytometry and imaging cytometry to detect endometrial cancer cells were 72.7% and 45.5%, respectively. Meanwhile, the overall specificities of flow cytometry and imaging cytometry for healthy controls were 75.0% and 81.3%, respectively. Our immuno-magnetic beads have very low auto-fluorescence, so they could be useful for fluorescent analysis, such as fluorescent immunochemical staining. In the future, these novel immuno-magnetic beads could be used for cytological study.

An efficient method of sorting liver stem cells by using immuno-magnetic microbeads

AIM： To develop a method to isolate liver stem cells fast and efficiently. METHODS： Fetal mouse liver cells were characterized by cell surface antigens （c-Kit and CD45/TER119） using flow cytometry. The candidate liver stem cells were sorted by using immuno-magnetic microbeads and identified by -lone-forming culture, RT-PCR and immunofluorescence says. RESULTS： The c-Kit-（CD45/TER119）- cell population with 97.9% of purity were purified by immuno-magnetic microbeads at one time. The yield of this separation was about 6% of the total sorting cells and the cell viability was above 98%. When cultured in vitro these cells had high clone-forming and self-renewing ability and expressed markers of hepatocytes and bile duct cells. Functionally mature hepatocytes were observed after 21 d of culture. CONCLUSION： This method offers an excellent tool for the enrichment of liver stem cells with high purity and viability, which could be used for further studies. It is fast, efficient, simple and not expensive.

Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

Overthelastseveraldecades,harmfulalgalblooms(HABs)havebecomeaseriousenvironmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.,) were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%-91.6% of the cells were captured from unialgal culture by IMBs, and only 5.3%-12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently in use.

Drug-eluting beads chemoembolization combined with programmed cell death 1 inhibitor and lenvatinib for large hepatocellular carcinoma

BACKGROUND The combination of transarterial chemoembolization(TACE),lenvatinib,and programmed cell death 1(PD-1)inhibitor has been widely used in the treatment of advanced hepatocellular carcinoma(HCC)and has achieved promising results.However,there are few studies comparing whether drug-eluting beads TACE(DTACE)can bring more survival benefits to patients with large HCC compared to conventional TACE(C-TACE)in this triplet therapy.AIM To compare the efficacy and adverse events(AEs)of triple therapy comprising DTACE,PD-1 inhibitors,and lenvatinib(D-TACE-P-L)and C-TACE,PD-1 inhibitors,and lenvatinib(C-TACE-P-L)in patients with large HCC(maximum diameter≥5 cm),and analyze the prognostic factors.METHODS Following a comprehensive review of our hospital’s medical records,this retrospective study included 104 patients:50 received D-TACE-P-L,and 54 received CTACE-P-L.We employed Kaplan-Meier estimation to assess the median progression-free survival(PFS)between the two groups,utilized Cox multivariate regression analysis to identify prognostic factors,and applied theχ2 test to evaluate AEs.RESULTS The objective response rate(ORR)and median PFS were significantly higher in the D-TACE-P-L group compared to the C-TACE-P-L group(ORR:66.0%vs 44.4%,P=0.027;median PFS:6.8 months vs 5.0 months,P=0.041).Cox regression analysis identified treatment option,portal vein tumor thrombus,and hepatic vein invasion as protective factors for PFS.AEs were comparable between the two CONCLUSION D-TACE-P-L may have significantly better PFS and ORR for large HCC,while exhibiting similar AEs to C-TACE-PL.

H_4SiW_(12)O_(40)/TiO_2/beads光催化降解亚甲基蓝的研究

以钛酸四丁酯为原料,空心微珠为载体,采用溶胶-凝胶法制备TiO2/beads光催化剂载体,然后浸渍法制备出H4SiW12O40/TiO2/beads表面负载修饰型复合光催化剂,并运用SEM、XRD、FT-IR和DRS对催化剂进行表征和分析。研究了H4SiW12O40/TiO2/beads对亚甲基蓝降解的光催化活性,考察了光强度、pH值、曝气量、底物浓度和催化剂用量等对催化效率的影响。实验结果表明,在中性条件下,H4SiW12O40/TiO2/beads催化剂的投加量为0.25 g/L,浓度为7.5 mg/L的亚甲基蓝溶液在250 W的紫外灯和600 W的可见光灯下光照60 min降解率分别可达到94.5%和55%。

溶胶-凝胶法制备TiO_2·SiO_2/beads及其光催化性能的研究

研究以四异丙醇钛 [Ti (iso OC3 H7) 4 ]、硅酸乙酯为原料 ,以空心玻璃微球为载体 ,用溶胶 凝胶法制备可漂浮附载型复合光催化剂TiO2 ·SiO2 /beads的过程 ,利用附载型复合光催化剂降解有机磷农药。结果表明 :附载型复合光催化剂活性显著提高 ,牢固性增强 ,TiO2 ·SiO2 摩尔比存在最佳值。n (TiO2 ) /m (SiO2 ) =30 / 70时 ,光催化剂活性最高 ,其活性是同样降解条件下 ,同样含量DegussaP 2 5TiO2 的

TiO_2·SiO_2/beads降解有机磷农药的研究

研究以四氯化钛 ,硅酸乙酯为原料 ,以空心玻璃微球为载体 ,用浸涂法制备TiO2 ·SiO2 /beads负载型复合光催化剂的过程。对利用负载型复合光催化剂降解有机磷农药进行了活性评价。结果表明 :TiO2 ·SiO2 /beads光活性显著提高 ,最佳光催化剂活性是同样降解条件下、同样含量的DegussaP - 2 5TiO2 光活性的 1.35倍左右。另外 ,对TiO2 ·SiO2 /beads的吸附性和比表面进行测试 ,同时从微观角度对负载型复合光催化剂的表面状态和光催化剂颗粒尺寸进行分析。

附载型复合光催化剂TiO_2·SiO_2/beads降解有机磷农药

研究以四异丙醇钛 [Ti( iso- O C3 H7) 4]、硅酸乙酯为原料 ,以空心玻璃微球为载体 ,用溶胶—凝胶法制备可漂浮附载型复合光催化剂 Ti O2 · Si O2 / beads的过程 ,利用附载型复合光催化剂降解有机磷农药。结果表明 ,复合型光催化剂 Ti O2 · Si O2 摩尔比存在最佳值 ,n ( Ti O2 ) / m ( Si O2 ) =30 / 70时 ,光催化剂活性最高 ,其活性是同样降解条件下、同样含量 Degussa P- 2 5Ti O2 的 2倍左右。该光催化剂比表面大 ,吸附性强。并用 XRD和

TiO_2·SiO_2/beads的制备与性质

研究以空心玻璃微球为载体 ,用Sol gel法制备附载型复合光催化剂的过程 ,并对TiO2 ·SiO2 /beads光催化活性进行评价 .结果表明 :复合型光催化剂TiO2 ·SiO2 摩尔比存在最佳值 ,当n(TiO2 ) /m(SiO2 ) =30 /70时 ,光活性是在同样降解条件下 ,同样含量DegussaP 2 5TiO2 的 2倍左右 .同时研究制备方法、制备条件、TiO2 ·SiO2 摩尔比、TiO2 ·SiO2 /beads吸附性、活化条件、比表面等因素对附载型复合光催化剂活性的影响 .

CoPcS/TiO_2/beads及TiO_2/beads光催化降解水面漂浮植物油

以钛酸四丁酯为原料,以空心玻璃微珠为载体,采用溶胶-凝胶法制备出TiO2/beads光催化剂,用浸渍法制备出CoPcS/TiO2/beads光催化剂。研究了TiO2/beads及CoPcS/TiO2/beads光催化降解水面漂浮植物油的最优条件。结果表明,酸性或中性条件下,375 W中压汞灯照射2～3 h,TiO2/beads与CoPcS/TiO2/beads的投加量分别为3 g与1 g时,植物油的去除率达90%以上,投加微量的H2O2,可大大提高两者的光催化去除率。

BACs-on-Beads技术对NIPT阳性结果的诊断价值

目的探讨应用BACs-on-Beads(Bo Bs)技术在无创DNA产前检测(NIPT)阳性结果中的诊断价值。方法对2015年1月至2016年6月因NIPT阳性结果在西京医院产前诊断中心进行羊水穿刺的129例孕妇进行Bo Bs检测和染色体核型分析。结果 Bo Bs共检出异常数80例,检出率62.02%。其中21三体55例、18三体10例、13三体1例、性染色体异常14例(X0 3例,XXX 3例,XXY 5例,XYY 2例,X0/XY嵌合1例),准确率依次为84.62%、71.43%、14.29%、41.18%。染色体核型分析结果与Bo Bs结果完全一致,确证了49例NIPT假阳性结果。结论 NIPT存在一定比例的假阳性结果,阳性病例必须进行进一步产前诊断,Bo Bs技术具有快速,简便,准确、高通量等优点,是快速确诊的有效手段。

BACs-on-Beads技术在快速诊断常见染色体异常中的应用

目的探讨细菌人工染色体标记-微球鉴别分离技术(BACs-on-Beads,BoBs)在快速诊断常见染色体异常中的应用。方法利用BoBs和染色体核型分析技术对856例孕妇的羊水标本进行检测,统计分析两种检测方法的一致性。结果856例孕妇共检出155例染色体异常病例,总体异常率为18.11%。两种方法同时检出141例染色体异常(包括136例非整倍体和5例嵌合体),染色体核型分析单独检出9例染色体异常(包括4例嵌合体和5例平衡易位),BoBs单独检出5例染色体微缺失/微重复。在染色体数目异常检出方面,BoBs和染色体核型分析的一致率为100%,对嵌合体检测的符合率为55.56%,对平衡易位和微缺失/微重复检测的符合率为0%。结论BoBs技术可以快速检测常见染色体数目异常及部分染色体微缺失/微重复,但存在漏诊风险。仅根据BoBs检测结果对胎儿作进一步处理存在一定风险,BoBs检测与染色体核型分析联合应用可以快速诊断常见染色畸形以减轻孕妇心理负担。

CoPcS／TiO2／beads及TiO2／beads光催化降解水面浮油

以钛酸四丁酯为原料,空心玻璃微珠(beads)为载体,采用溶胶-凝胶法制备出TiO2/beads光催化剂,用浸渍法制备出CoPcS/TiO2/beads新型光催化剂.研究了利用TiO2/beads及CoPcS/TiO2/beads光催化剂降解水面漂浮植物油的最优条件.结果表明:(1)溶胶-凝胶法制备TiO2/beads的最佳条件为:空心玻璃微珠浸渍3次,450～550℃下焙烧2 h.用CoPcS对TiO2/beads进行改性时,TiO2/beads的最佳浸渍时间为30 min.(2)在中性或酸性条件下,375 W中压汞灯光照2～3 h,TiO2/beads与CoPcS/TiO2/beads的最佳用量分别为3 g和1 g(植物油3 g),在此条件下,植物油的去除率都高达90%.(3)微量H2O2对TiO2/beads和CoPcS/TiO2/beads的光催化活性都有很大的提高.对于TiO2/beads催化剂,H2O2的最佳用量为5～11 mmol/L;对于CoPcS/TiO2/beads催化剂,H2O2的最佳用量为5～30 mmol/L.(4)新型光催化剂CoPcS/TiO2/beads比TiO2/beads具有更好的除油性能.

Pb(Ⅱ) biosorption using chitosan and chitosan derivatives beads: Equilibrium, ion exchange and mechanism studies

The study examined the adsorption of Pb（II） ions from aqueous solution onto chitosan, chitosan-GLA and chitosan-alginate beads. Several important parameters influencing the adsorption of Pb（II） ions such as initial pH, adsorbent dosage and different initial concentration of Pb（II） ions were evaluated. The mechanism involved during the adsorption process was explored based on ion exchange study and using spectroscopic techniques. The adsorption capacities obtained based on non–linear Langmuir isotherm for chitosan, chitosan-GLA and chitosan-alginate beads in single metal system were 34.98, 14.24 and 60.27 mg/g, respectively. However, the adsorption capacity of Pb（II） ions were reduced in the binary metal system due to the competitive adsorption between Pb（II） and Cu（II） ions. Based on the ion exchange study, the release of Ca2＋, Mg2＋, K＋ and Na＋ ions played an important role in the adsorption of Pb（II） ions by all three adsorbents but only at lower concentrations of Pb（II） ions. Infrared spectra showed that the binding between Pb（II） ions and the adsorbents involved mostly the nitrogen and oxygen atoms. All three adsorbents showed satisfactory adsorption capacities and can be considered as an efficient adsorbent for the removal of Pb（II） ions from aqueous solutions.

Effects of Polyethylene Oxide Concentration on the Size of Beads in Electrospun Beaded Nanofibers

Embedding particle drugs in beaded nanofibers by electrospinning has been shown a potential approach to control drug release in tissue engineering. The bead size is one of the critical parameters in controlling the drug release rate. In this study,the relationship between polymer concentration and beads size was investigated. Aqueous polyethylene oxide( PEO) solutions with different concentrations were prepared to obtain various beaded nanofibers by electrospnning. Optical microscope and scanning electron microscope( SEM) were used to observe the variation tendency of bead size. With an increase in the polymer concentration,the diameter of fibers between beads became bigger,while the fiber uniformity improved. In addition, the polymer concentration influenced the distribution of bead diameter. Higher polymer concentration would reduce the possibility of small-sized beads formation and improve the uniformity of bead diameter. The study provides a possible way to control the size of beads,which is helpful for further research on the control of particle drug release.

Efficacy of intra-arterial contrast-enhanced ultrasonography during transarterial chemoembolization with drug-eluting beads for hepatocellular carcinoma

AIM To assess the usefulness of intra-arterial contrastenhanced ultrasonography(IAUS) during transarterial chemoembolization(TACE) with drug-eluting beads(DEB) for hepatocellular carcinoma(HCC).METHODS Thirty two patients with 39 HCC underwent DEB-TACE guided with IAUS, and examined by contrast-enhanced ultrasonography(CEUS) or dynamic CT after DEB-TACE were enrolled in this study. CEUS findings before DEBTACE and IAUS findings were compared. Treatments judged to be complete and incomplete for lesions were appropriate and insufficient, respectively. Findings on CEUS and/or dynamic CT performed 1, 3 and 6 mo after DEB-TACE were evaluated using m RECIST(CR/PR/SD/PD).RESULTS The treatments were complete and incomplete in 26 and 13 lesions, respectively. On imaging evaluation using CEUS and/or dynamic CT one month after treatment, 25 and 1 lesions were judged to be CR and PR, respectively, and at 6 mo after treatment, the results were CR, PR, SD and PD for 24, 1, 0 and 1 of these lesions, respectively, in the 26 completely treated lesions. Of the 13 lesions in which treatment was incomplete, the results on imaging at one month after treatment were CR, PR, SD and PD for 0, 6, 4 and 3 lesions, respectively. The overall CR rate at 6 mo after treatment was 61.5%(24/39).CONCLUSION A combination of DEB-TACE with IAUS can improve the therapeutic effects in patients with HCC.

Highly efficient generation of GGTA1 knockout pigs using a combination of TALEN m RNA and magnetic beads with somatic cell nuclear transfer

The transcription activator-like effector nuclease （TALEN） technique combined with the somatic cel nuclear transfer （SCNT） method has been successfuly applied for creating geneticaly modiifed pigs. However, methods for isolating cels with bialelic indels requires further improvement because of the relatively low enrichment efifciency of mutated somatic cels. Moreover, little is known regarding the off-target effects of the TALEN system and the heredity of TALEN-modiifed pigs. In this study, an efifcient method to increase the enrichment efifciency of TALEN-mediated bialelic knockout （KO） cels was established, and corresponding geneticaly modiifed pigs with the expected genotype were generated whose off-target effect, fertility and heredity characteristics were aslo evaluated. Two TALEN pairs were constructed to target the porcine α-1,3-galactosyltransferase （GGTA1） gene locus. TALEN mRNA was transfected into the ear ifbroblasts folowed by the enrichment of α-Gal nul cels of minipigs using isolectin B4 （IB4） lectin and magnetic beads. A total of 115 cel colonies were formed and validated to beGGTA1 KO cels by sequencing and 10 bialelic KO cel colonies were used as nuclear donors for SCNT. ThirtyGGTA1 bialelic KO piglets were successfuly delivered and grew normaly. Seventeen potential off-target sites were investigated, and no off-target events were detected in the live piglets. To determine the fertility and heredity characteristics of TALEN-modiifed pigs, 10 mature founders were mated with each other and the mutations were determined to be transmitted to the F1 piglets. We established a robust and safe technology for developing geneticaly modiifed pig lines with expected genotypes for agricultural breeding and biomedical application.

Nine-year experience of doxorubicin-eluting beads chemoembolization for hepatocellular carcinoma

BACKGROUND: Chemoembolization with doxorubucin-eluting beads (DEB) has been used to treat hepatocellular carcinoma (HCC) since 2007. This study compared the efifcacy and sur-vival between transarterial chemoembolization (TACE) with DEB and conventional approach (cTACE) in HCC treatment. METHODS: This retrospective case-control study compared the overall survival and tumor response of HCC patients to cTACE (n=190) and DEB (n=143) by the reassessment of com-puted tomography and serum alpha-fetoprotein (AFP). Mul-tivariate analysis was used to determine the factors affecting tumor response. RESULTS: The median post-treatment to pre-treatment AFP level was 0.8 for a DEB session (n=258) and 1.0 for a cTACE session (n=452), showing a signiifcantly greater decrease in AFP after DEB (P<0.05). More patients in the DEB group achieved objective response (complete and partial) compared with those in the cTACE group (P<0.05). Objective tumor re-sponse after DEB vs cTACE was 34.8% vs 15.4% in 0-3 months (P=0.001), 37.1% vs 20.0% in 3-6 months (P<0.05), and 50.0%vs 30.0% in 6-12 months (P=0.093). DEB predicted a 3.604 times odds of achieving at least one objective tumor response in a patient when compared to cTACE (P<0.0001). The median survival from ifrst transcatheter therapy of patients having&nbsp;undergone at least once DEB was 12.53 months, while those having received cTACE only was 10.53 months (P=0.086). A tendency of improved survival appeared to maintain until >80 months after the ifrst TACE session in the DEB group. CONCLUSION: DEB is a safe alternative to cTACE in HCC pa-tients with better therapeutic efifcacy.

Microsatellite Enrichment from AFLP Fragments by Magnetic Beads

A simple and efficient method was presented for isolating microsatellite DNA markers from peanut (Arachis hypogaea L.) genome. The genomic DNA was converted into pre-amplified AFLP fragments and hybridized with biotin-labeled SSR probes. Then the hybrid mixture was used to incubate with magnetic beads coated with streptavidin. After washing to remove the non-SSR fragments, the eluted single-strand DNA, which was cloned and sequenced, was largely enriched for microsatellites. Primers can then be designed according to the sequence flanking the repeat motifs and used for polymorphism analysis. The whole experiment can be completed within one week and can be employed as a reliable option for any molecular laboratory to develop SSR markers.