胃肠道间质瘤生物学特性临床病理分析

目的: 探讨胃肠道间质瘤的临床病理特点,对预后做出客观评价.方法: 对46例胃肠道间质瘤的形态学特征、生物学特性、免疫组化特点进行分析,并对部分病例作临床随访.结果: 胃肠道间质瘤胃比肠道多发,5 a生存率为61.9%,属低度恶性肿瘤.免疫组化表型为CD34 ,CD117 .结论: 胃肠道间质瘤的良恶性不能依据形态学指标定,肿瘤大小和核分裂数、肿瘤性坏死在判断良恶性时有重要参考价值.     

Beta2-microglobulin removal and plasma albumin levels with high cut-off hemodialysis

PURPOSE: beta2-microglobulin (beta2MG) is pivotal to the pathogenesis of dialysis-related amyloidosis. We compared the effects of high cut-off hemodialysis (HCO-HD) with those of standard high-flux hemodialysis (HF-HD) regarding the concentration and clearance of beta2MG and albumin. DESIGN: We enrolled ten patients with acute renal failure in a double-blind, cross-over, randomized controlled trial. PROCEDURES: Each patient received four hours of HCO-HD (estimated in vivo cutoff 50-60 kDa) and four hours of HF-HD (estimated in vivo cutoff 15-20 kDa) in random order. Statistical methods and outcome measures: As data lacked normal distribution, we used nonparametric statistical analysis. Plasma and dialysate concentrations of beta2MG and albumin were measured at baseline and after four hours of each study treatment. MAIN FINDINGS: We found significantly greater diffusive beta2MG clearances for HCO-HD compared to HF-HD (at the start: 71.8 ml/min vs. 5.1 ml/min; P=0.008 and at the end: 68.8 ml/min vs. 5.7 ml/min; P=0.008). We found a reduction in plasma beta2MG concentrations of -31.6% during HCO-HD compared to an increase by 25.7% during HF-HD; P=0.008. At baseline (HCO-HD: 26.0 g/L vs. HF-HD: 26.5 g/L), and at the end of both treatments, plasma albumin concentrations were comparable (HCO-HD: 25.5 g/L vs. HF-HD: 26.5 g/L; P=0.25). During HCO-HD, albumin clearance was 1.9 ml/min at the start and decreased significantly to 0.8 ml/min at the end; P=0.008. HF-HD had an albumin clearance of 0.01 ml/min. CONCLUSIONS: HCO-HD was more effective in decreasing plasma beta2MG concentrations than standard HF-HD and did not reduce plasma albumin levels. Further studies of HCO-HD in the treatment of dialysis-related beta2MG accumulation appear warranted.     

骨髓间充质干细胞移植对急性心肌梗死大鼠血管内皮功能的影响

目的:观察骨髓间充质干细胞移植治疗急性心肌梗死对血管内皮功能的影响。方法:实验于2005-09/2006-06在唐山工人医院中心实验室完成。①选用清洁级健康近交系SD大鼠40只,随机数字表法分为假手术组、模型对照组、干细胞移植组、细胞培养基组,10只/组。另取1只大鼠用于骨髓间充质干细胞的提取。②大鼠处死后无菌条件下分离股骨和胫骨,剔除肌肉、骨膜,剪一侧骨端,用10mL无血清DMEM低糖培养基冲出骨髓细胞后,收集冲洗液制成单细胞悬液,过滤离心,弃上清液,密度梯度法分离培养骨髓间充质干细胞。待细胞80%贴满培养瓶底时,用乙二胺四乙酸和胰蛋白酶混合消化传代。将4,6-二脒-2-苯基吲哚以50mg/L浓度加入第3代细胞培养基中,制成细胞悬液备用。③术前各组大鼠均行气管插管,建立急性心肌梗死模型,以远端供血区心肌组织颜色苍白、心电图检测Ⅱ导联ST段持续抬高为模型建立成功的标志。假手术组仅开胸予以前降支穿线但不结扎。④造模成功后1～3h,干细胞移植组用微量注射器吸取4,6-二脒-2-苯基吲哚标记好的第3代骨髓间充质干细胞悬液50μL,直接注入梗死区边缘的心肌组织。细胞培养基组按干细胞移植组的方法于相同部位注射等量无血清DMEM低糖培养基(pH值6.9)。⑤造模后4周处死各组大鼠,切取心脏组织,于梗死边缘区心肌组织制作切片,荧光显微镜下观察4,6-二脒-2-苯基吲哚标记的供体细胞。留取2mL全血,离心后抽取血清500μL,酶联免疫吸附法测定血清中细胞间黏附分子1与血管内皮细胞黏附分子1的含量。结果:40只大鼠均进入结果分析。①体外培养骨髓间充质干细胞的性状观察:从正常大鼠骨髓中分离培养的原代骨髓间充质干细胞,24～48h贴壁生长,短棒状;7～10d达到80%～90%融合;传至第3代骨髓间充质干细胞分布均匀,呈纺锤形。②各组心脏标本4,6-二脒-2-苯基吲哚标记移植细胞的存活状况:干细胞移植组可见成团及散在的4,6-二脒-2-苯基吲哚标记的阳性细胞,其余3组心脏标本中均未见荧光细胞。③各组大鼠血清中细胞间黏附分子1与血管内皮细胞黏附分子1的含量检测:与假手术组比较,模型对照组、细胞培养基组、干细胞移植组细胞间黏附分子1与血管内皮细胞黏附分子1的含量均明显升高(P<0.01或0.05);与模型对照组比较,干细胞移植组细胞间黏附分子1与血管内皮细胞黏附分子1的含量均有所降低[(3.37±0.14),(2.10±0.15)μg/L;(2.64±0.13),(1.74±0.06)μg/L;P均<0.05]。结论:骨髓间充质干细胞移植到实验性大鼠心肌梗死模型后成功存活,能降低血清中增高的细胞间黏附分子1与血管内皮细胞黏附分子1含量,提示骨髓间充质干细胞移植可能通过降低黏附分子的表达来改善血管内皮功能。     

2，4，6-三硝基苯磺酸诱导小鼠炎症性肠病模型的建立

目的:建立2,4,6-三硝基苯磺酸诱导的结肠炎动物模型,并分析其剂量效应。方法:实验于2006-05/09在南方医院消化内科研究所完成,选用SPF级雌性Balb/c小鼠50只,按随机数字表法分为5组,每组10只,对照组予以300g/L乙醇溶液灌肠1次;其他4组分别给予含25mg/kg,50mg/kg,100mg/kg,150mg/kg2,4,6-三硝基苯磺酸的300g/L乙醇溶液灌肠1次。各组于造模后3,7,21,28d各处死1只小鼠,观察结肠的大体形态。造模后21d观察各组小鼠结肠的病理组织切片,并检测中性粒细胞的髓过氧化物酶活性。结果:造模过程中50,100,150mg/kg2,4,6-三硝基苯磺酸组各死亡2,3,3只小鼠。①各组小鼠造模后的一般情况:对照组及25mg/kg2,4,6-三硝基苯磺酸组小鼠发生一过性的结肠机械损伤表现,出现便血等现象,但3～7d后开始缓解;而50～150mg/kg2,4,6-三硝基苯磺酸组小鼠出现便血,腹泻。②各组小鼠结肠的大体形态及病理组织学观察结果:对照组和25mg/kg2,4,6-三硝基苯磺酸组小鼠肠道在第7天时表现为无粘连,局部充血,肠壁不增厚,未见溃疡,于第3周时局部充血已好转;结肠病理切片大致正常。50～150mg/kg2,4,6-三硝基苯磺酸组小鼠肠道肉眼可见粘连,肠腔增大,充血水肿,溃疡形成,出现肠道增厚,病理切片见黏膜下层水肿,固有层炎症细胞浸润等现象,以上现象于造模4周后开始缓解。其中当2,4,6-三硝基苯磺酸剂量为100mg/kg时作用达到高峰。③各组小鼠的中性粒细胞髓过氧化物酶活性比较:50,100,150mg/kg2,4,6-三硝基苯磺酸组均显著高于对照组[(14.17±3.33,18.50±2.33,18.17±1.50,4.83±2.00)nkat/L,(P<0.01)]。结论:应用含100mg/kg2,4,6-三硝基苯磺酸的300g/L乙醇溶液灌肠是诱导小鼠结肠炎较为理想的方法。     

Pharmacokinetic studies of recombinant human insulin-like growth factor I (rhIGF-I)/rhIGF-binding protein-3 complex administered to patients with growth hormone insensitivity syndrome

CONTEXT: GH insensitivity syndrome (GHIS), Laron syndrome, is characterized by severe short stature, high serum GH levels, and very low serum IGF-I and IGF-binding protein-3 (IGFBP-3) levels associated with a genetic defect of the GH receptor. Recombinant human (rh) IGF-I treatment at doses of 80-120 microg/kg given sc twice daily is effective in promoting growth in these patients. We have investigated a newly developed drug, rhIGF-I/rhIGFBP-3, a 1:1 molar complex of rhIGF-I and rhIGFBP-3. OBJECTIVES: The objectives of the study were to determine IGF-I pharmacokinetics after the administration of rhIGF-I/rhIGFBP-3 in adolescents with GHIS and to evaluate its safety and tolerability. DESIGN: This was an open-label clinical study. SETTING: The study was conducted in a general pediatric ward of a university teaching hospital. PARTICIPANTS: Four patients (one female and three males; mean age, 14.9 yr; mean height sd score, -4.9) with confirmed molecular diagnosis of GHIS agreed to participate in the study. INTERVENTION: rhIGF-I/rhIGFBP-3 was administered in a single sc injection at 0.5 and 1.0 mg/kg.dose (equivalent to 100 and 200 microg/kg rhIGF-I) after breakfast with a 2-d interval between doses. RESULTS: IGF-I levels reached a maximum between 19 +/- 8.3 and 15 +/- 6.2 h for the low and high doses, respectively. The circulating IGF-I levels obtained with the low and high doses were similar, although a discrete dose-dependent increase in circulating IGF-I levels was observed. The IGF-I half-life in four subjects after a dose of 0.5 mg/kg rhIGF-I/rhIGFBP-3 was estimated to be 21+/- 4 h. There were no acute adverse events reported, and all blood glucose measurements were normal. CONCLUSION: These data demonstrated that the rhIGF-I/rhIGFBP-3 complex was effective in increasing levels of circulating total and free IGF-I into the normal range for a 24-h period after a single sc administration in patients with GHIS, and that administration of rhIGF-I/rhIGFBP-3 was safe and well tolerated.     

Outcome following patent ductus arteriosus ligation in premature infants: a retrospective cohort analysis

Background  

The patent ductus arteriosus (PDA) is an important problem in premature infants. Surgical PDA ligation is usually only be considered when medical treatment has either failed or was contraindicated. The aims of our study were to determine the mortality and morbidity following patent ductus arteriosus ligation in premature infants, and whether prostaglandin synthetase inhibitor (PSI) use prior to ligation affects outcome.     

Okadaic acid inhibits relaxant neural transmission in rat gastric fundus in vitro

The aim of the present study was to characterize the influence of the phosphatase type 1 and 2A inhibitor okadaic acid on non-adrenergic, non-cholinergic (NANC) neurotransmission in the rat gastric fundus. Okadaic acid (10-6 M), an inhibitor of protein phosphatases 1 and 2A, did not show any influence on the basal tonus or on a contraction plateau induced by 5-HT (10-7 M) within 30 min of observation. When okadaic acid (10-6 M) was applied 10 min prior to 5-HT (10-7 M), the contraction plateau of serotonin was unchanged. To investigate the inhibitory neurotransmission, the muscle strips were pre-contracted using 5-HT (10-7 M), and inhibitory stimuli were applied at the contraction plateau, which was stable over 30 min. The inhibitory effects of vasoactive intestinal peptide (VIP), nitric oxide (NO) and electrical field stimulation (EFS, 40 V, 0.5 ms, frequencies ranging from 0.5 to 16 Hz) were examined. When okadaic acid (10-6 M) was applied prior to EFS-induced NANC relaxation, significant attenuation of the inhibitory response was demonstrated (16 Hz: control: -92.4 +/- 1.9%; okadaic acid 10-7 M: -60.7 +/- 6.1%; okadaic acid 10-6 M: -25.3 +/- 3.4%; n=11; P < 0.01). By contrast, neither the concentration-dependent inhibitory actions of VIP (10-11-10-8 M) (VIP 10-8 M: -100%; VIP 10-8 M + okadaic acid 10-6 M: -89.9 +/- 8.3%; n=8; n.s) nor that of diethylamine nitric oxide (DEA-NO) (3 x 10-7-10-4 M) (DEA-NO 10-4 M: -95.3 +/- 8.4%; DEA-NO 10-4 M + okadaic acid 10-7 M: -98.3 +/- 6.3%; DEA-NO 10-4 M + okadaic acid 10-6 M: 96.5 +/- 7.6%; n=9; n.s.) on 5-HT induced contraction were altered by pre-incubation with okadaic acid (10-6 M). This is the first report that supports the concept that protein phosphatases 1 and 2A may contribute to the regulation of rat gastric fundus motility. The protein phosphatase inhibitor okadaic acid significantly reduces electrically induced inhibitory NANC responses, while leaving direct muscular effects of the inhibitory NANC neurotransmitters VIP and NO unaffected - suggesting a neural site of action. The potential roles of protein phosphatases on NANC neurotransmission remain to be clarified in detail, as this might offer a new pathway for modulating smooth-muscle function.     

Greater numbers of human spermatozoa associate with endosalpingeal cells derived from the isthmus compared with those from the ampulla

A simple co-culture bioassay system was used to investigate whether or not the anatomical origin affected the ability of epithelial cells from the human uterine (Fallopian) tube to 'bind' spermatozoa. This study was also used to identify some of the factors which may be involved in the regulation of sperm-epithelial interactions in vitro by comparing different tissue culture models and assessing the effect of oestradiol concentration. Epithelial explants harvested from different regions of human uterine tubes were co-incubated with a known concentration of motile donor spermatozoa. All results were adjusted to reflect a standard sperm concentration of 5 x 10(6)/ml. More spermatozoa associated per field of isthmic compared to ampullary epithelium [isthmus 9.5 +/- 0.9, ampulla 7.1 +/- 0.7 (mean +/- SEM); n = 36, P < 0.05, ANOVA] and cells from post-menopausal patients had an apparently reduced ability to bind spermatozoa [isthmus 5.5 +/- 2.0, ampulla 4.3 +/- 1.4 (mean +/- SEM); n = 4]. Neither menstrual cycle stage nor addition of mid-cycle concentrations of 17beta-oestradiol (750 pmol/l) affected the number of spermatozoa which bound to epithelium from either tubal region. In addition, the number of spermatozoa which bound per field of polarized explants was greater (P < 0.05) than that bound to dissociated primary and passaged epithelial cell monolayers. This report is the first to provide evidence suggestive of a role for sperm- epithelial binding in the formation of an isthmic sperm reservoir in the human uterine tube. Results also indicate that oestrogen is not involved in the regulation of these interactions, and that cell polarity is an important factor for such associations in vitro.