分离培养汗腺导管部细胞的新方法

目的 探讨建立汗腺导管部细胞分离的新技术.方法 成人仞厚皮片和薄中厚皮片标本(n=10)剪碎后用Ⅱ型胶原酶消化12 h,吸取并转移汗腺导管到培养皿中贴壁培养.应用流式细胞仪、免疫组织化学染色和逆转录-聚合酶链式反应(RT-PCR)以及蛋白印迹(Western Blot)分析检测培养细胞的汗腺特异标志CEA、CK8、CK18、CK19抗原表达,并用膜片钳技术检测培养细胞膜上阿米洛利(amiloride)敏感Na~+离子通道,用t检验比较分析两组间实验数据.结果 汗腺导管贴壁48 h后,围绕汗腺导管出现单层扁平的上皮细胞,生长2～4周融合成片.流式细胞学检查示原代培养汗腺导管细胞与原代培养汗腺细胞在癌胚抗原(CEA)阳性率[(90.26±1.12)%vs.(89.70±1.43)%]和细胞角蛋白8(CK8)阳性率[(94.41±1.84)%vs.(93.65±1.63)%]上,差异无统计学意义(P>0.05).形态学染色汗腺导管细胞抗CEA、CK8、CK18、CK19染色均为阳性.RT-PCR表明原代培养汗腺导管细胞表达CEA、CK8、CK18、CK19基因,Western Blot清晰显示CEA条带,CK8、CK18、CK19蛋白条带.膜片钳检测表明原代培养汗腺导管细胞膜上存在amiloride敏感Na~+离子通道.无血清表皮细胞EpiLife培养基在汗腺导管细胞生长过程中抑制成纤维细胞生长.结论 从仞厚皮片和中厚皮片分离培养汗腺导管部细胞的方法较传统的分离方法具有简便快速的优点,EpiLife培养基可抑制培养过程中成纤维细胞的生长,可以在体外建立最佳汗腺导管细胞模型.     

蛋白A免疫吸附联合利妥昔单抗在PRA阳性肾移植受者中的应用七例报告

目的 探讨术前群体反应性抗体(PRA)阳性的肾移植受者应用蛋白A免疫吸附联合利妥昔单抗(RTX)治疗的效果及安全性.方法 7例中,行二次肾移植者5例,有多次妊娠史者2例.7例均行亲属活体供肾移植,供者均为男性.供、受者ABO血型相同,补体依赖细胞毒试验阴性,HLA抗原错配数2～5个.术前单纯PRAⅠ类抗体增高者2例,单纯PRAⅡ类抗体增高者2例,PRA Ⅰ、Ⅱ类抗体均增高者3例;IgG平均为13.4 g/L,IgA平均为1.78 g/L,IgM平均为1.47 g/L.术前行免疫吸附2～10次,平均为5次.当患者PRA或免疫球蛋白水平降至正常值及以下时进行移植,并于手术当天单次静脉输注RTX 600 mg,同时给予抗淋巴细胞球蛋白或抗胸腺细胞球蛋白.术后采用他克莫司(或环孢素A)、霉酚酸酯及泼尼松预防排斥反应.监测患者血肌酐及内生肌酐清除率(Ccr);临床上考虑患者发生排斥反应时,行移植肾穿刺活检;分别于术前及术后1周测定患者外周血中CD19+和CD20+细胞百分比(仅检测3例).结果 免疫吸附及RTX治疗期间,患者均未出现不良反应.行免疫吸附后,7例患者的PRA Ⅰ、Ⅱ类抗体均有所下降,IgG、IgA和IgM的平均值也有所下降.术后患者移植肾功能恢复良好,未发生移植肾功能恢复延迟,其中3例分别在术后8、10和14 d出现急性排斥反应,治疗后均逆转.术前患者外周血中CD19+细胞分别为1.98%、4.64%和4.45%,CD20+细胞分别为15.15%、2.01%和1.15%;术后1周时,其外周血中CD19+细胞分别为0.39%、0.98%和0.11%,CD20+细胞分别为0.34%、0.15%和0.01%.术后1例发生肺部巨细胞病毒感染,治疗后好转,1例因肺部烟曲霉感染而死亡,死亡时肾功能正常.结论 对于术前PRA阳性者应用蛋白A免疫吸附及RTX治疗有一定效果,经处理后患者可接受肾移植.     

烧伤创面水疱液对人骨髓间充质干细胞体外培养及表型转化的影响

目的 了解烧伤水疱液对人骨髓间充质于细胞(MSC)体外培养及表型转化的影响.方法 体外分离、培养、扩增、鉴定人骨髓MSC,收集烧伤患者伤后12、24、48 h水疱液,分别加入MSC培养液中.分为水疱液1组:MSC培养液中加入伤后12 h水疱液;水疱液2组:MSC培养液中加入伤后24 h水疱液;水疱液3组:MSC培养液中加入伤后48 h水疱液;对照组:常规培养.各组所加水疱液体积分数均为20%.观察水疱液微生物生长情况及各组MSC生长情况.流式细胞仪检测培养8 d时各组MSC中CD44、细胞角蛋白7(CK7)阳性表达率.结果 15个水疱液标本细菌、真菌培养均为阴性;各组MSC细胞形态无明显变化,但各水疱液组细胞数量均少于对照组,水疱液3组细胞数量下降最明显.水疱液1、2、3组CD44阳性表达率分别为(83.0±3.1)%、(77.2±2.9)%、(65.1±2.3)%,均低于对照组[(89.5±3.2)%,P<0.01];水疱液1、2、3组CK7阳性表达率分别为(24.06±0.11)%、(16.41±0.09)%、(4.48±0.07)%,明显高于对照组[(3.87±0.04)%,P<0.01].结论 烧伤创面水疱液对MSC体外培养有明显的抑制作用,并具有一定的促进MSC表型转化作用.     

A comparison of plasma levobupivacaine concentrations following transversus abdominis plane block and rectus sheath block

Levobupivacaine is commonly used as the local anaesthetic of choice in peripheral nerve blocks, but its pharmacokinetics have not been fully investigated. We compared the changes in plasma concentrations of levobupivacaine following transversus abdominis plane block and rectus sheath block. Fifty woman undergoing laparoscopy were randomly allocated to receive either a transversus abdominis plane block or an rectus sheath block. In both groups, 2.5 mg.kg^?1 levobupivacaine was administered, and blood samples were obtained 15 min, 30 min, 60 min and 120 min after injection. The mean maximum plasma concentration (Cmax) and mean time to reach Cmax (Tmax) as determined by non‐linear regression analysis were 1.05 μg.ml^?1 and 32.4 min in the transversus abdominis plane group and 0.95 μg.ml^?1 and 60.9 min in the rectus sheath group, respectively. The plasma concentration of levobupivacaine peaked earlier in the transversus abdominis plane group than in the rectus sheath group and the maximum plasma concentration depended on the dose administered but not the procedure.     

颈椎椎弓根三种置钉法的实验研究

目的: 评价 3种颈椎椎弓根置钉方法。方法: 3组尸体各 9具, 行颈椎CT影像测量后分别用 3种方法进行椎弓根置钉, 其结果由CT验证。结果: 颈椎椎弓根三维定位器法置钉 90枚, 椎弓根内占 90%, 其中神经根损伤 2枚, 椎动脉损伤 1枚; Abumi法置钉 90枚, 椎弓根内占 55. 56%, 其中神经根损伤 19枚, 椎动脉损伤 4枚; Ebraheim法置钉 72枚, 椎弓根内占 29. 17%, 其中神经根损伤 24枚, 椎动脉损伤 29枚, 3种方法均无脊髓损伤。结论: 在术中不探查椎弓根、无影像设备导向条件下, 颈椎椎弓根三维定位器置钉准确率较Abumi手法置钉高, 而Ebraheim法不适用于国人。     

Sentinel node biopsy in squamous cell cancer of the oral cavity and oral pharynx: a diagnostic meta-analysis

BACKGROUND: The sentinel node biopsy concept has been gaining support in the head and neck cancer literature during only the last few years, and several pilot studies have been published. This procedure aims to avoid unnecessary treatment to the clinically negative neck by identifying the patients with occult neck disease. METHODS: We performed a systematic review and a diagnostic meta-analysis of all published literature regarding sentinel node biopsies in head and neck cancer until December 2003 using established guidelines. Using the pooled sensitivity rates obtained from the meta-analysis and treatment outcomes from published literature, we created a decision analysis model to identify the treatment arm with better payoffs. RESULTS: A total of 301 patients with oral cavity primary tumors and 46 patients with oropharyngeal primary tumors from 19 articles were included for the meta-analysis. The pooled sensitivity result using the random effects model was 0.926 (95% confidence interval, 0.852-0.964). The cumulative payoff for the sentinel node biopsy arm was lower than that for the elective node dissection arm by about 1%. The payoffs were assigned for the recurrence and mortality rates only and did not take into account the morbidity caused by the procedures. CONCLUSIONS: The sentinel node biopsy procedure has shown high sensitivity rates in pilot studies for oral and oropharyngeal squamous cell cancer across the globe and is reliable and reproducible. This study provides a firm evidence base for forthcoming trials on the role of sentinel node biopsy in head and neck cancer.     

Hollow fiber-based liquid phase microextraction followed by analytical instrumental techniques for quantitative analysis of heavy metal ions and pharmaceuticals

Hollow-fiber liquid-phase microextraction(HF-LPME)and electromembrane extraction(EME)are miniaturized extraction techniques,and have been coupled with various analytical instruments for trace analysis of heavy metals,drugs and other organic compounds,in recent years.HF-LPME and EME provide high selectivity,efficient sample cleanup and enrichment,and reduce the consumption of organic sol-vents to a few micro-liters per sample.HF-LPME and EME are compatible with different analytical in-struments for chromatography,electrophoresis,atomic spectroscopy,mass spectrometry,and electrochemical detection.HF-LPME and EME have gained significant popularity during the recent years.This review focuses on hollow fiber based techniques(especially HF-LPME and EME)of heavy metals and pharmaceuticals(published 2017 to May 2019),and their combinations with atomic spectroscopy,UV-VIS spectrophotometry,high performance liquid chromatography,gas chromatography,capillary elec-trophoresis,and voltammetry.     

实时剪切波弹性成像在评估子宫内膜容受性中的初步应用

背景　子宫内膜容受性是在胚胎着床前,内膜能够使胚胎正常着床的一种综合情况。近年来,超声对子宫内膜容受性的研究成为热点,但既往多倾向于对内膜厚度、回声和血流的研究,随着超声新技术的发展,子宫内膜弹性有望成为内膜容受性研究的新方向。目的　利用实时剪切波弹性成像技术对子宫内膜进行定量测量,对比分析容受期与增殖期子宫内膜弹性模量值的差异,探讨实时剪切波弹性成像在评估子宫内膜容受性中的应用价值。方法　选择2018年1—12月在温州市人民医院门诊行卵泡监测的备孕妇女80例,分别在内膜增殖期和容受期对其行二维超声和实时剪切波超声弹性成像检查,获取并比较两个时相的内膜回声类型、内膜厚度以及内膜平均弹性模量值,进行相关性分析。结果　80例女性的子宫内膜在增殖期主要表现为均匀低回声的三线征,容受期主要表现为均匀高回声。增殖期和容受期内膜厚度和内膜平均弹性模量值分别为7.0（6.0,8.0）mm、（25.7±4.4）kPa及11.0（10.0,12.8） mm、（9.4±2.7）kPa。容受期内膜厚度高于增殖期,容受期内膜平均弹性模量值低于增殖期（P<0.05）。内膜厚度与内膜平均弹性模量值之间呈负相关（rs=-0.849,P<0.05）。不同回声类型内膜平均弹性模量值比较,差异有统计学意义（P<0.05）。结论　实时剪切波弹性成像能定量评估子宫内膜弹性,间接反映内膜时相,结合内膜厚度及回声类型,能为不孕症患者内膜容受性的评估提供新的诊断思路。     

大学英语词汇教学探索

本文着重强调词汇在大学英语教学中的重要性,探讨了大学英语词汇教学的内容及核心,并且根据自己的教学实践,阐述了有效的词汇教学方法不仅能极大促进学生对词汇的记忆和掌握,同时还有助于提高学生其他方面的能力.     

呼出气冷凝液生物标志物检测在呼吸系统疾病中的应用现状及前景

呼吸系统疾病越发受到人们广泛关注,其复杂多变的病征给医疗领域带来一系列挑战,许多临床常用的检查方法已不足以应对该局面,临床亟需更加方便快捷的检查方法为诊疗提供准确有效的依据。呼出气冷凝液（EBC）是以无创方式获得的呼吸道内衬液,是呼吸系统疾病生物标志物的一种新的重要来源,具有较高的临床研究价值。基于EBC的生物标志物检测方法,能有效反映患者病情变化,优化治疗方案。该方法以其操作简单、安全无创、重复性高、依从性好等优点,日益受到临床重视。本文系统分析了EBC中生物标志物检测方法在呼吸系统疾病中的诊断、监测、疗效评价及预后中的应用现状,并对其收集、检测规范及临床应用等提出相应问题,为其在临床上广泛应用提供有力依据。