摘要：The sieving and enrichment of rare tumor cells from large-volume pleural effusion(PE)samples is a promising technique for cell-based lung tumor diagnosis and drug tests,which features high throughput and recovery,purification,as well as viability rates of rare target cells as the prerequisites for high sensitivity,specificity,and accuracy of tumor cell *** this paper,we propose a three-dimensional(3 D)sieving method for rare tumor cell enrichment,which effectively eliminates the"dead zones"in traditional two-dimensional(2 D)cell filters with a dimension-raising strategy to satisfy the requirements mentioned *** prototype device was combined with a funnel-shaped holder,a flexible micropore membrane in the middle,and a3 D spiral fluid channel covered on the membrane as a three-layer ice-creaming cone composite *** by gravity alone,the device performed as follows:(1)20-fold throughput compared with the 2 D commercial planee hich was up to 20 mL/min for a threefold dilution of whole blood sample;(2)high recovery rates of 84.5%±21%,86%±25%,83%±14%for 100,1000,and 10000 cells/mL,respectively,in 30 mL phosphate buffer saline(PBS)sample,and a 100%positive detection rate in the case of≤5 A549 cells in 1 mL PBS;(3)a typical purification rate of 85.5%±9.1%;and(4)a viability rate of>93%.In the demonstration application,this device effectively enriched rare target cells from large volumes(>25 mL)of clinical pleural *** following results indicated that tumor cells were easy-to-discover in the enriched PE samples,and the proliferation capability of purified cells was(>4.6 times)significantly stronger than that of unprocessed cells in the subsequent 6-day *** above evaluation indicates that the proposed easily reproducible method for the effective execution of rare cell enrichments and assays is expected to become a practical technique for clinical cell-based tumor diagnosis.

摘要：Fungi are nature’s recyclers,allowing for ecological nutrient cycling and,in turn,the continuation of life on *** fungi inhabit the human microbiome where they can provide health benefits,while others are opportunistic pathogens that can cause ***,members of the fungal kingdom,have been domesticated by humans for the production of beer,bread,and,recently,medicine and ***,the great untapped potential exists within the diverse fungal ***,many yeasts are intractable,preventing their use in biotechnology or in the development of novel treatments for pathogenic ***,as a first step for the domestication of new fungi,an efficient DNA delivery method needs to be ***,we report the creation of superior conjugative plasmids and demonstrate their transfer via conjugation from bacteria to 7 diverse yeast species including the emerging pathogen Candida *** create our superior plasmids,derivatives of the 57 kb conjugative plasmid pTA-Mob 2.0 were built using designed gene deletions and insertions,as well as some unintentional ***,a cluster mutation in the promoter of the conjugative gene traJ had the most significant effect on improving conjugation to *** addition,we created Golden Gate assembly-compatible plasmid derivatives that allow for the generation of custom plasmids to enable the rapid insertion of designer genetic ***,we demonstrated that designer conjugative plasmids harboring engineered restriction endonucleases can be used as a novel antifungal agent,with important applications for the development of next-generation antifungal therapeutics.

摘要：β-Glucuronidase(GUS)plays a key role in tumor initiation,metastasis,and progression,and thus,has been proposed as a promising cancer *** this study,we designed an enzyme-activatable near-infrared(NIR)fluorescent probe(DCM-βGlcA)for the rapid and accurate detection of GUS activity in vitro,in vivo and ex *** DCM-βGlcA was prepared by linking a glucuronic acid residue to dicyanomethylene-4 H-pyran(DCM).This probe exhibited significant light-up NIR fluorescent signals at 680 nm after reacting with GUS and the Stokes shift could reach 150 *** DCM-βGlcA showed a high sensitivity toward GUS and an excellent linear relationship at concentrations ranging between 0 and 4 U L^(-1)(R^(2)=0.9974)with the limit of detection as low as 0.19 U L^(-1).We used the DCM-βGlcA to identify GUS serum levels in both cancer patients and healthy individuals with a similar accuracy as that of an enzyme-linked immunosorbent assay(ELISA)while being easier and faster to ***,the DCM-βGlcA was used for tracking endogenous GUS in living cells,thereby discriminating GUSoverexpressed liver cancer from normal ***,the DCM-βGlcA was able to detect and image endogenous GUS in liver cancer tissue and tumor-bearing mouse *** findings demonstrate the potential of the DCM-βGlcA as a promising tool for detecting and monitoring GUS activity in preclinical applications.

摘要：背景:儿童如果发生结核病,通常会假设其传染源是生活在同一家庭的成年传染性病例。本研究应用限定性片断长度多态性分析技术(RFLP)来确定从传染源到儿童的传播。设计:收集1993年到1998年诊断的成人及儿童结核病例和他们的结核分枝杆菌培养物。通过访视和结核分枝杆菌菌株的RFLP分析来确定流行病学联系及证实家庭内传播。结果:1291名成人中有1139名(91％)和417名儿童中有65名(16％)籍培养结果证实为结核病。由于要重新收集痰标本和提取DNA的问题,仅对其中的832名成人和35名儿童进行了RFLP分析:19名(54％)儿童有家庭成员确诊为结核病,12名儿童 有与其家庭成员相同的菌株。分离自儿童的29株(83％)菌株构成了社区簇的一部分,但与传染源有明确接触史者只有15株。结论:在具有传染性成人肺结核病例的家庭中,儿童患有结核病并不意味着必然是成人对儿童的传播。儿童可能是在社区也可能是在家庭中受到感染。此结果对在高发病率地区的接触者追踪及治疗策略有意义。