Influence of medium components on ex vivo megakaryocyte expansion

Reinfusion of ex vivo-expanded autologous megakaryocytes together with a stem cell transplantation may be useful to prevent or reduce the period of chemotherapy-induced thrombocytopenia. In this study, we analyzed several serum-containing and serum-free media to identify the most suitable medium for megakaryocyte expansion. Moreover, two thrombopoietin (Tpo)-mimetic peptides were tested to evaluate whether they could replace Tpo in an expansion protocol. To analyze the effects of different media on megakaryocyte expansion, we used an in vitro liquid culture system. For this purpose, CD34(+) cells were isolated from peripheral blood and cultured for 8 days in the presence of Tpo and interleukin-3 (IL-3). The presence of megakaryocytes was analyzed by flow cytometric analysis after staining for CD41 expression. For our standard culture procedure, megakaryocyte medium (MK medium) supplemented with 10% AB plasma was used. Addition of 5% or 2.5% AB plasma yielded higher numbers of megakaryocytes, implying the presence of inhibitory factors in plasma. However, some plasma components are required for optimal megakaryocyte expansion because addition of less than 1% AB plasma or addition of human serum albumin instead of AB plasma resulted in the formation of lower numbers of megakaryocytes. Two commercially available serum-free media were also tested: Cellgro and Stemspan. If CD34(+) cells were cultured in Cellgro medium similar numbers of megakaryocytes were obtained as when CD34(+) cells were cultured in MK medium supplemented with 10% AB plasma. In MK medium with 2.5% AB plasma, higher numbers of megakaryocytes were cultured than in MK medium supplemented with 10% AB plasma. Therefore, Cellgro medium is not the best alternative medium. In cultures with Stemspan medium, higher numbers of megakaryocytes were obtained compared to MK medium with 10% AB plasma. Stemspan is thus a good alternative for MK medium. Two Tpo-mimetic peptides, AF13948 and PK1M, were tested for their ability to replace Tpo. In cultures with AF13948, comparable numbers of megakaryocytes were obtained as in the presence of Tpo, but in cultures with PK1M the number of megakaryocytes was lower. This study shows that high concentrations of plasma in medium inhibits megakaryocyte formation, but some plasma components are required for optimal megakaryocyte expansion. For an ex vivo expansion protocol, it is worthwhile to test several media, because the number of megakaryocytes differs widely with the medium used.     

十二指肠-胃反流物成分对胃黏膜的影响

目的:探讨原发性十二指肠胃反流(DGR)是否造成胃黏膜的病理性损害以及十二指肠液中各种成分与动态pH的变化和胆红素吸收值(ABS)的关系。方法:应用带胃管的便携式pH监测仪和Bilitec2000,同步监测14例原发性病理性DGR患者和15名健康志愿者的胃内24h胆汁反流和pH变化情况。根据胃内pH和ABS的对应变化,实时抽吸胃液送检,测出胃液中胆汁酸(BA)、总胆红素(TBIL)、淀粉酶(AMY)和脂肪酶(LIP)的浓度。结果:ABS与TBIL、BA以及pH值与AMY浓度呈正相关。重度胃炎组的BA高于轻、中度组。白天胃液中AMY和LIP分别高于夜间;而夜间的BA、TBIL和DBIL较高。结论:胃腔内的pH值变化主要是胰液反流而非胆汁反流所致。胃黏膜的损伤程度主要与BA的反流有关,胰液反流对胃黏膜的损伤弱于胆汁反流。胃腔中TBIL可以作为胆汁反流性胃炎的监测指标,而不是诱发或导致胃炎的因素。     

Influence of posture on hepatic perfusion and the presystemic biotransformation of propranolol: simulation of the food effect

Several research groups have reported that the oral administration of propranolol with protein-rich food leads to a marked increase (mean + 60%) in the area under the drug plasma concentration-time curve (AUC oral) of this highly metabolized and well-absorbed drug. It has been postulated that this "food effect" is caused at least in part by a transient increase in hepatic blood flow (QH) with its associated decrease in first-pass metabolism (hepatic extraction is a monotonic decreasing function of QH). A randomized crossover study using postural manipulations to produce changes in QH of the magnitude observed after food consumption (20% to 50%) was performed in an attempt to isolate the contribution of transient changes in QH to the food effect phenomenon. A solution of 80 mg propranolol HCl was taken orally and subjects were randomly assigned to postural manipulation protocols that should change QH such that AUC oral would be minimized (phase 1) or maximized (phase 2). Estimated QH (indocyanine green total body clearance from blood) was determined before and at three time points during each phase. It was observed that indocyanine green total body clearance during periods of standing was 15% to 40% below that observed during periods of seating (significant at p less than 0.05 for many of the appropriate comparisons). However, AUC oral for propranolol was not affected (mean +/- 1 SD; AUC phase 2/AUC phase 1+= 0.98 +/- 0.28) by these changes in QH, which are comparable to those encountered after food consumption.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of seasonal biorhythms on urinary excretion of enzymes and other parameters

Urinary excretion of enzymes, electrolytes, creatinine, protein and the consumption of food and water in female and male Wistar rats was examined over two seasons of summer and wintertime, respectively. The evaluation of data revealed that gamma-glutamyltransferase, inorganic phosphate, and the lysosomal enzymes, N-acetyl-beta-D-glucosaminidase and acid phosphatase, were significantly different between the collection periods in both seasons. Other parameters showed significant differences only in male rats, whereas urinary electrolytes, with the exception of chloride, showed no significant differences.     

职业紧张对ICU男护士血清抗氧化物酶的影响

[目的]探讨职业紧张对ICU男护士血清抗氧化酶的影响。[方法]采用整群抽样法抽取某省16所三级甲等综合医院的126名ICU男护士,采用职业紧张量表调查男护士的职业紧张感,并监测其血清抗氧化物酶的水平,分析职业紧张对其产生的影响。[结果]ICU男护士职业紧张量表中的职业任务(ORQ)与个体紧张反应(PSQ)评分均高于国内常模(P0.05);个人应对资源(PRQ)评分低于国内常模(P0.05)。不同得分水平组间比较差异有统计学意义(P0.05)。[结论]通过对ICU男护士职业紧张感进行调查,并研究其对血清抗氧化酶的影响,提示医院管理者应关注男护士这一特殊的群体,并采取有针对性的干预措施,从而提高护理质量,以顺应优质护理服务的开展。     

代谢综合征及其组分对踝臂指数的影响

目的调查健康体检人群中代谢综合征（MS）及其各组分数量对踝臂指数（ABI）的影响,为MS并发心血管疾病的健康管理提供科学依据。方法对2007—2009年在我院进行体检的公职人员7819例个体进行随机抽样,采用中华医学会糖尿病学分会关于中国人MS的建议诊断MS,对样本进行了MS组分数量分级。结果①比较非MS组与MS组的ABI指标,MS组患者ABI值降低,分别为1.11±0.07和1.09±0.12,P〈0.05;PAD患病率增加,分别为0和7.0%,差异有统计学意义（P〈0.05）。②MS组分别与M0、M1、M2比较,随MS组分数目的增加,ABI值逐渐下降,PAD患病率逐渐上升,差异有统计学意义（P〈0.05）。③按性别分组后对MS各组分与ABI相关性研究,结果发现血脂异常、高血压、高血糖组分的PAD患病率较高,女性各MS组分PAD患病率均高于男性。结论北京市健康体检人群中MS及其组分对ABI存在影响,ABI随MS组分的增加而降低,且MS患者的PAD发病率高于非MS,血脂异常、高血压、高血糖均对PAD存在影响。     

Influence of hormone replacement therapy on blood antioxidant enzymes in menopausal women

BACKGROUND: Natural loss of estrogen occurring in menopausal process may contribute to various health problems many of them possibly related to oxidative stress. Hormone replacement therapy (HRT) is the most common treatment to attenuate menopausal disturbances. This study was aimed at evaluating the influence of HRT on the activity of antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; and glutathione peroxidase, GPx) and lipid peroxidation (thiobarbituric acid reactive substances, TBARS) in menopausal women. METHODS: Blood antioxidant enzyme activities were determined in premenopausal (n=18) and in postmenopausal healthy women without (n=21) or with (n=19) HRT (mean ages: 47, 59, and 57 years, respectively). RESULTS: TBARS, CAT, and GPx activity were not significantly different among the groups of study. However, SOD activity was significantly lower in postmenopausal women without HRT (0.68+/-0.04 U/mg Hb) when compared both to premenopausal women (0.91+/-0.04 U/mg Hb) and to postmenopausal women with HRT (0.89+/-0.07 U/mg Hb). SOD activity was positively correlated to the duration of HRT in the postmenopausal groups (r=0.33, p<0.05). CONCLUSIONS: HRT antagonizes the decrease of SOD activity that occurs after menopause, suggesting that HRT may play a beneficial role in the protection against oxidative stress.     

围产期窒息对新生儿心肌酶学和肾功能的影响

目的探讨围产期窒息对新生儿心肌酶学和肾功能的影响。方法对90例围产期窒息患儿进行脐动脉血心肌酶谱、新生儿外周血心肌酶谱和肾功能进行测定。结果围产期窒息患儿脐血和外周血肌酸激酶(CK)和肌酸激酶同工酶(CK-MB)均较正常新生儿的显著升高(P<0.05),且外周血的升高更明显(P<0.01)。围产期窒息患儿外周血尿素氮和肌酐明显升高,且肾功能受损程度与窒息程度成正比(P<0.05)。早产窒息儿较足月窒息儿更易发生肾功能损伤(P<0.05)。结论围产期窒息患儿早期进行心肌酶学和肾功能检测,对相关异常早诊断、早治疗,对疾病预后的判断及治疗具有重要意义。     

cDNA cloning of biotransformation enzymes belonging to the cytochrome P450 family in the antennae of the noctuid moth Mamestra brassicae

The involvement of cytochrome P450 enzymes in olfaction was demonstrated in vertebrates some time ago. In insects these enzymes are well known for their role in insecticide resistance, but the involvement of P450 in pheromone degradation was only recently demonstrated. Using a PCR strategy, we have isolated two cDNAs from the antennae of the cabbage armyworm Mamestra brassicae - CYP4L4 and CYP4S4 - which encode microsomal P450s. CYP4S4 expression is restricted to the antennae, whereas CYP4L4 is also found in the proboscis and legs. Moreover, the two genes are strongly expressed in one type of sensory unit of the antennae - the sensilla trichodea - which are tuned to the detection of odourants. The putative function of the corresponding enzymes is discussed with regard to their respective expression patterns.     

Influence of blood components on the tissue uptake indices of cyclosporin in rats

The effect of blood components on the transfer of cyclosporin into brain, salivary gland, liver and kidney was measured in rats by an an vivo double isotope, tissue-sampling single injection technique. The brain-, salivary gland- and liver extraction of [3H] cyclosporine relative to [14C]butanol were studied with an intracarotid and a portal vein injection technique; the kidney extraction of [3H]cyclosporin relative to p[14C]aminohippuric acid was measured after a rapid injection in the aorta. The injection vehicles were a Ringer's solution [Ringer-4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid], rat plasma, human plasma, human red blood cells (RBC) suspension and human blood. The brain extraction (1-4%), the salivary extraction (25-74%) and the kidney extraction (8-47%) varied markedly depending on which blood components were added to the injection solution. The effect of RBC binding on tissue extraction was by far more pronounced than that of plasma protein binding. The binding of cyclosporin to RBC retards particularly the uptake of this drug by kidney. Moreover the kidney penetration of cyclosporin was confirmed by thaw-mount autoradiography after injection of the labeled drug in the aorta; the micrographs showed that the renal clearance of cyclosporin was restricted largely to the glomerular route. In contrast the hepatic extraction of [3H]cyclosporin was high (48-84%) and generally nonlimited by its binding to blood components; this could explain the high concentrations of drug observed in this tissue. In summary, cyclosporin binding to RBC and plasma proteins reduces drug uptake by brain, kidney and salivary gland, but causes little inhibition of hepatic uptake of cyclosporin.