海马区星形胶质细胞培养上清液体外诱导人脂肪基质细胞向神经元样细胞的分化

目的:观察海马区星形胶质细胞培养上清液能否在体外诱导人脂肪基质细胞向神经元样细胞分化。方法:实验于2004-10/2005-06在华北煤炭医学院中心实验室完成。在无菌条件下从Wistar乳鼠分离出海马组织,从分离的海马组织中获得星形胶质细胞,并收集其培养上清液。取外科手术获得的人腹部皮下脂肪组织进行人脂肪基质细胞的原代培养。30例患者均知情同意。取第3代人脂肪基质细胞接种到培养孔中,预先放置无菌盖玻片的24孔培养板,制备细胞爬片或者接种到培养瓶中,细胞生长达50%～60%融合时,去除培养液,换为海马区星形胶质细胞培养上清诱导液进行诱导,对照组培养液为无血清培养基。倒置相差显微镜下连续观察细胞生长情况和形态变化,应用免疫细胞化学、鉴定神经前体细胞的特异性标志神经巢蛋白、神经细胞的特异性标志神经元特异性烯醇化酶、微管联合蛋白2和神经胶质细胞的特异性标志胶质纤维酸性蛋白的表达。结果:①诱导培养第3天,部分人脂肪基质细胞开始变形,从原先的细长梭状细胞变成神经元样细胞,可见细胞伸出突起,多为双极或多极细胞。②刚分离接种的人脂肪基质细胞镜下呈圆形,悬浮状态,接种后24h内贴壁,并开始伸展,多呈梭形。1周后细胞融合成单层,排列出现方向性,但有少量圆形及卵圆形细胞混杂生长。③第4,5代人脂肪基质细胞在诱导48h后形态即开始发生变化,扁平的胞体较预诱导后逐渐回缩,向外伸出突起,72h后扁平的胞浆向胞核收缩,突起继续延长,以后随时间进展,具有典型神经细胞形态特点的细胞数量逐渐增多,形成双极或多极细胞。④免疫细胞化学检测人脂肪基质细胞诱导5d后发现有(10.5±3.7)%神经巢蛋白、(38.4±5.2)%胶质纤维酸性蛋白、(15.7±2.3)%神经元特异性烯醇化酶表达,未见微管联合蛋白2的表达。结论:海马区星形胶质细胞培养上清液可以在体外诱导人脂肪基质细胞向神经元样细胞方向分化。     

Management strategies for human babesiosis

ABSTRACT

Introduction

Human babesiosis is reported throughout the world and is endemic in the northeastern and northern Midwestern United States and northeastern China. Transmission is primarily through hard bodied ticks. Most cases of severe disease occur in immunocompromised individuals and may result in prolonged relapsing disease or death.     

A pilot study for one-stop endoscopic total extraperitoneal inguinal hernia repair

Background  

One-stop surgery was developed for patients to undergo surgical evaluation, anesthesia, surgery, and discharge all within 1 day. This study aimed to assess the feasibility, patient satisfaction, and potential of one-stop endoscopic total extraperitoneal (TEP) inguinal hernia surgery.     

In vitro susceptibility testing of Borrelia burgdorferi sensu lato isolates cultured from patients with erythema migrans before and after antimicrobial chemotherapy

Clinical treatment failures have been reported to occur in early Lyme borreliosis (LB) for many suitable antimicrobial agents. Investigations of possible resistance mechanisms of the Borrelia burgdorferi complex must analyze clinical isolates obtained from LB patients, despite their receiving antibiotic treatment. Here, borrelial isolates obtained from five patients with erythema migrans (EM) before the start of antibiotic therapy and again after the conclusion of treatment were investigated. The 10 isolates were characterized by restriction fragment length polymorphism analysis and plasmid profile analysis and subjected to susceptibility testing against a variety of antimicrobial agents including those used for initial chemotherapy. Four out of five patients were infected by the same genospecies (Borrelia afzelii, n = 3; Borrelia garinii, n = 1) at the site of the EM lesion before and after antimicrobial therapy. In one patient the genospecies of the initial isolate (B. afzelii) differed from that of the follow-up isolate (B. garinii). No significant changes in the in vitro susceptibilities became obvious for corresponding clinical isolates before the start and after the conclusion of antimicrobial therapy. This holds true for the antimicrobial agents used for specific chemotherapy of the patients, as well as for any of the additional agents tested in vitro. Our study substantiates borrelial persistence in some EM patients at the site of the infectious lesion despite antibiotic treatment over a reasonable time period. Borrelial persistence, however, was not caused by increasing MICs or minimal borreliacidal concentrations in these isolates. Therefore, resistance mechanisms other than acquired resistance to antimicrobial agents should be considered in patients with LB resistant to treatment.     

Use of a water-soluble busulfan formulation--pharmacokinetic studies in a canine model

In a canine model we investigated the toxicity and pharmacokinetics of a water soluble busulfan preparation. Busulfan was dissolved in dimethylsulfoxide (DMSO) and administered either orally or intravenously in a single dose of 1 mg/kg. The application in either preparation was well tolerated. In seven dogs, peak levels in the range of 730 ng/mL to 1,000 ng/mL were measured after intravenous injection with an area under curve (AUC) of 75 ng.h/kg.mL to 146 ng.h/kg.mL. It was of note that even the oral administration of the same busulfan preparation resulted in AUC values in the same range as observed after parenteral application. The absorption rate of busulfan tablets in our model was as unpredictable as documented in clinical trials. On the basis of the present study, clinical trials using busulfan dissolved in DMSO given either intravenously or orally appear warranted. This approach should lead to predictable blood levels, reduced toxicity, and increased efficacy of busulfan-containing regimens.     

Identification of kallikrein and FXIa as impurities in therapeutic immunoglobulins: implications for the safety and control of intravenous blood products

Background and Objectives The occurrence of thromboembolic events (TEEs) with intravenous immunoglobulin lots (IVIGs) raised the question of the causative agent for these adverse events. We investigated the predominant plasma proteases in 19 IVIG lots from five manufacturers including three lots associated with adverse events. Material and Methods The inhibitor profile of the amidolytic activity in IVIG lots was investigated with substrates S‐2302 and S‐2288. In immunocapture assays, prekallikrein and FXI antigen and respective active proteases were quantified. Non‐activated partial thromboplastin time (NAPTT) and a modified FXIa PTT served as global and FXIa‐specific clotting assays, respectively. Results Kallikrein was identified as one major contaminant activity in IVIGs. A second activity was seen in some IVIGs with substrate S‐2288, but not with S‐2302. Inhibition studies excluded FXIIa, thrombin or plasmin as contaminant activity. FXI antigen was seen in all 19 IVIG lots, and FXIa activity was found as second major impurity in some IVIGs, including all lots involved in TEEs. FXIa highly correlated with a short clotting time in NAPTT. Conclusions Kallikrein and FXIa are the major contaminants in IVIGs. FXIa was highly procoagulant, with highest level in TEE‐associated IVIGs. Since the NAPTT unambiguously identified FXIa procoagulant activity in IVIGs, its implementation as a release test would improve the safety of IVIGs.     

新疆和田地区碘缺乏病流行区儿童听力调查分析

和田地区不仅是新疆,而且是世界上最严重的碘缺乏区之一.由于碘的缺乏,1987年自治区组成碘缺乏病流行区预防小组,由各行专家,对严重碘缺乏地区的初期妊娠妇女、出生后1个月婴儿、1岁儿童、2岁儿童分成4组,进行口服补碘及土地灌溉加碘防治工作.     

Hydrosalpingeal fluid inhibits in-vitro embryonic development in a murine model

Recent evidence describing a suboptimal clinical outcome in women with hydrosalpinges who undergo in-vitro fertilization (IVF) and embryo transfer suggests a potential deleterious effect of this fluid on in- utero embryo development. Consequently, we evaluated in-vitro mouse embryo development in the presence of hydrosalpingeal fluid (HF) collected from 10 infertile women of reproductive age. Chemical analyses showed both similarities and differences of these fluids to reported values for fluids collected from non-diseased Fallopian tubes. The HF had a significant deleterious effect upon mouse embryo cleavage and development to the expanded and hatched blastocyst stage, although the effect was variable among patients. Dilution of HF to 30% concentration with culture medium failed to negate this effect. This argues against the effect resulting from a relative lack of critical, supportive component(s) in the HF. Additionally, further experiments performed with cultures under an oil overlay significantly reduced the embryotoxicity of the HF. This evidence suggests there may be a lipophilic factor that can impair embryo development. The relatively poor IVF-embryo transfer success in women with proximally patent hydrosalpinges may be explained, at least in part, by reflux of a lipophilic embryotoxic factor(s) into the uterine cavity.