Abnormal dentin structure in two novel gene mutations [COL1A1, Arg134Cys] and [ADAMTS2, Trp795-to-ter] causing rare type I collagen disorders

Histological and ultrastructural observations of dentin of two patients affected with rare types of type I collagen disorders are presented. In the first case, a homozygous nonsense mutation in ADAMTS2 (substitution of a codon for tryptophan by a stopcodon) causes type VIIC Ehlers-Danlos syndrome (EDS) with multiple tooth agenesis and focal dysplastic dentin defects. In the second case, a missense mutation in COL1A1 (substitution of arginine by cysteine) results in a type I EDS phenotype with clinically normal-appearing dentition. Tooth samples are investigated by using light microscopy (LM), transmission electron microscopy (TEM) and immunostaining for types I and III collagen, and tenascin. These are compared with samples from patients with types III and IV osteogenesis imperfecta (OI) in association with dentinogenesis imperfecta (DI), showing a consistently abnormal appearance of the dentin in all specimens, with variations being primarily those of degree of change. Similarities in histological changes include the alternating presence of normal and severe pathological areas in primary and secondary dentin, the latter being characterized by large canal-like structures in atubular areas. Ultrastructural evidence of pathological dentinogenesis include abnormal distribution, size and organization of collagen fibers, which may also be found in clinically unaffected teeth. The histological and ultrastructural changes seen can be explained on the basis of odontoblast dysfunction which may be secondary to the collagen defect, interfering with different levels of odontoblast cell function and intercellular communication. These observations on (ultra)structural dentin defects associated with the two novel gene mutations are the first ever reported.     

人NT-proBNP的克隆及原核表达

目的：克隆人氨基末端脑钠肽（amino—terminal pro—brain natriuretic peptide,NT—proBNP）基因,构建原核表达载体并纯化表达产物。方法：采用PCR从正常成人cDNA中扩增NT—proBNP基因,将其克隆至pGEM—T中,测定其核苷酸序列。然后构建原核表达载体pGXE4T-2-NT-proBNP,用IPTG诱导表达,GSH—agarose亲和纯化蛋白。结果：经PCR扩增获得NT—proBNP基因,测序正确,在大肠杆菌中融合表达后,该蛋白的表达量占菌体总蛋白的20％,用SDS—PAGE和Western blot鉴定,显示其相对分子量34600。经亲和纯化后的GST—NT—proBNP的纯度可以达到96％,得率为1．8mg／100ml。结论：NT—proBNP的纯化成功,为建立NT—proBNP检测方法奠定了基础。     

体外循环血液净化新技术治疗急危重症患者的研究进展

肝衰竭、肾衰竭、成人呼吸窘迫症、心力衰竭、急性肺损伤、内皮细胞功能衰竭以及脑损伤等是临床常见的危急重症,此类疾病多伴有血流动力学不稳定、器官低灌注等病理生理改变.体外循环血液净化技术是近年来救治上述危急重症患者的重要方法.该文针对体外循环血液净化新技术的作用原理、作用方式、应用类型,以及其对危急重症患者营养支持、药物治疗、调节酸碱平衡等治疗中的作用和意义进行分析,并通过查阅相关文献、综合目前国内外资料,总结体外循环血液净化技术的发展前景和应用意义.     

Vitreous hemorrhage and fibrovascular proliferation after laser-induced chorioretinal venous anastomosis

AIM: To describe a case in which vitrectomy was required for vitreous hemorrhage and fibrovascular proliferation after laser-induced chorioretinal venous anastomosis (LCVA) for non-ischemic central retinal vein occlusion (CRVO). METHODS: Observational case report. RESULTS: A 72-year-old man complained of central scotoma in the left eye, and was diagnosed as suffering from non-ischemic CRVO. LCVA was performed in another hospital. Although favorable visual function was briefly maintained postoperatively,severe vitreous hemorrhage developed in his left eye, necessitating vitrectomy. CONCLUSION: Considering that LCVA carries a risk of serious complications, we must apply this treatment with caution, especially in ethnic groups, such as the Japanese, in whom pigmentation reacts to photocoagulation excessively.     

脑卒中患者中检测 N - 末端脑钠肽前体、D 二聚体、超敏 C 反应蛋白的临床价值简

目的探讨在脑卒中患者中检测N-末端脑钠肽前体（NT-proBNP）、D二聚体（D-D）、超敏C反应蛋白（hs-CRP）的临床价值。方法选择101例脑卒中住院患者和100例健康对照组分别测定NT-proBNP、D-D、hs-CRP并分别进行比较。结果NT-proBNP、D-D和hs-CRP在梗死1组、梗死2组、梗死3组与对照组比较或两两比较中均有显著差异（P〈0．05）；在急性期、亚急性期、慢性期与对照组比较或两两比较中也均有显著差异（P〈0．05）。结论NT-proBNP、D-D和hs-CRP可作为预测脑卒中病程的三项指标，三者的联合检测可作为脑梗死面积大小和病程进展情况的参考。     

分泌型凋亡相关蛋白1调控成纤维细胞凋亡的研究

目的探讨分泌型凋亡相关蛋白1（secretedapoptosis-relatedprotein1,SARP1）调控增生性瘢痕患者的成纤维细胞（fibroblast,FB）凋亡的作用。方法构建SARP1腺病毒载体（Ad-sARPl）．感染瘢痕患者的皮肤FB,促进其表达SARP1蛋白,观察其蛋白表达后人FB增殖与凋亡的变化,明确SARP1蛋白对FB的调控作用,并通过转移酶介导的三磷酸脱氧鸟苷-生物素刻痕末端标记（TuNEL）方法、流式细胞仪（FACs）分析细胞增殖与凋亡动态变化。结果成功构建Ad-SARP1,并能够感染人FB,通过逆转录一聚合酶链式反应（RT-PCR）检测到SARP1的mRNA值明显增加,Western印迹方法检测SARP1蛋白表达明显增多（P〈0．05）;其蛋白表达后四甲基偶氮噻唑蓝（MTT）法检测,与带荧光的腺病毒空载体（Ad-EGFP）及对照组相比,Ad-SARP1感染的FB增殖活力均显著增高;TUNEL检测Ad-SARP1感染FB前后细胞凋亡结果显示,细胞凋亡明显受到抑制,凋亡阳性细胞变少,接近瘢痕成纤维细胞（HSFB）凋亡;FACS分析表明,感染组FB的细胞凋亡指数与对照组相比明显下降（P〈0．01）。结论sARP1参与调控增生性瘢痕患者的FB功能,抑制细胞凋亡,促进细胞增殖加快。     

Effect of adolescent exposure to MDMA and cocaine on acquisition and reinstatement of morphine-induce CPP

It is well known that an elevated percentage of ecstasy users also consume cocaine. Recently, it has been reported that a high frequency of heroin smokers first consumed heroin under the effects of ecstasy with the hope of reducing the stimulant effects of the latter drug. The aim of the present study was to evaluate the effect of exposure to MDMA and cocaine during adolescence on morphine-induced conditioned place preference (CPP) and reinstatement in adulthood. In the first experiment, adolescent mice were exposed to six injections of MDMA and three weeks later their response to the reinforcing properties of 40 mg/kg of morphine was evaluated using the CPP paradigm. All the treatment groups developed the same magnitude of morphine-induced preference and, after CPP was extinguished, it was restored in all the groups with a priming dose of 10 mg/kg of morphine. Only mice that had been treated with 10 or 20 mg/kg of MDMA had their morphine-induced preference reinstated after receiving only 5 mg/kg of morphine. In the second experiment, adolescent mice were similarly treated with six administrations of cocaine (25 mg/kg) or cocaine plus MDMA (5, 10 or 20 mg/kg), and their response to morphine-induce CPP was evaluated three weeks later. Similarly to the first experiment, all the groups developed a preference for the morphine-paired compartment, but this preference was not reinstated with a priming dose of 10 mg/kg of morphine following extinction, as was the case among the control animals. These results lead us to hypothesize that periadolescent MDMA exposure alters responsiveness to the rewarding properties of morphine, highlighting MDMA as a gateway drug whose use may increase the likelihood of dependence on other drugs.     

Regulation of ω-3 Fish Oil Emulsion on the SIRS during the Initial Stage of Severe Acute Pancreatitis

The aim of this study was to explore the effects of parenteral supplementation with ω-3 fish oil emulsion （Omegaven） on systemic inflammatory response syndrome （SIRS） during the initial stage of severe acute pancreatitis （SAP）. In a prospective, randomized and controlled trial, 60 patients with SAP were randomized either to treat with conventional therapy （Con group, n=30） or conventional therapy plus intravenous supplementation with ω-3 fish oil emulsion 0.2 g/kg every day （FO group, n=30）. The effects were analyzed by the SIRS-related indexes. The results showed that APACHE-Ⅱ scores in FO group were significantly lower, and the gap increased much farther after the 4th day than those in Con group （P〈0.05）. Fluid equilibrium time became shorter markedly in FO group than in Con group （5.1±2.2 days vs 8.4±2.3 days）. In FO group, SIRS scores were markedly decreased and the SIRS state vanished after the 4th day; Plasma level of TNF-α was significantly reduced, while IL-10 decreased markedly, most prominently between the 4th and 7th day, and the ratio of IL-10/TNF-α raised as compared with Con group （P〈0.05）. During the initial stage of SAP, parenteral supplementation with ω-3 fish oil emulsion could efficiently lower the magnitude and persistence time of the SIRS, markedly retrieve the unbalance of the pro-/anti-inflammatory cytokines, improve severe condition of illness and may provide a new way to regulate the SIRS.     

疏血通注射液治疗原发性肾病综合征的临床研究

目的探讨疏血通注射液治疗原发性肾病综合征（NS）的临床疗效及对血浆高敏c反应蛋白（hs—CRP）的影响。方法将确诊为原发性Ns的130例患者随机分为治疗组和对照组,所有患者均给予激素等常规治疗,治疗组68例患者同时加用疏血通注射液4～6ml加入5％葡萄糖溶液100～250ml中静脉滴注,每天1次,15d为1个疗程,连用1—2个疗程。观察两组患者的临床疗效、不良反应以及治疗前后血浆hs—CRP水平的变化。结果治疗组总缓解率为89．71％,与对照组（70．97％）比较差异有统计学意义（P〈0．05）,疏血通注射液能够降低血浆hs—CRP水平,经比较差异具有统计学意义（P〈0．05）,且未出现明显不良反应。结论疏血通注射液治疗原发性NS临床疗效满意,具有抗炎作用,安全性好。     

(Pro-)Insulin Biosynthesis and Release of Newly Synthesized (Pro-)Insulin from Isolated Islets of Rat Pancreas in the Presence of Amino Acids and Sulphonylureas+,++

Abstract. The influence of arginine, lysine, tolbutamide and glibenclamide on (pro-)insulin biosynthesis and release of newly synthesized (pro-)insulin was studied in isolated islets of rat pancreas. Islets were incubated with ³H-leucine and glucose in the presence and absence of the test agents. Proinsulin and insulin of islets and incubation media were separated by gel filtration on Sephadex G 50. Estimations were carried out for radioactivity and immunoreactivity for insulin. All four test substances were able to enhance insulin release whereas no stimulation of leucine incorporation into (pro-)insulin was found. Arginine and tolbutamide even markedly reduced (pro-)insulin synthesis. Conversion of proinsulin to insulin was not affected by any of the test agents. For studying the influence of the 4 substances on secretion of newly synthesized (pro-)insulin two experimental models were used: 1) Labelling of the islets in the presence of the test agents, followed by uniform stimulation with glucose alone in the presence or absence of Ca⁺⁺. 2) Addition of the 4 test substances after uniform prelabelling of the islets. 1) Presence of arginine and sulfonylureas during labelling resulted in a significantly enhanced relative fractional release of newly synthesized (pro-)insulin, although the bulk of secreted hormone appeared to stem from the storage pool also under these conditions. The enhanced fractional release was persistent also during the postlabelling period when the islets had been labelled in the presence of arginine or glibenclamide. On the other hand, a decreased release of newly synthesized (pro-)insulin was observed during the postlabelling period in islets labelled in the presence of tolbutamide. Lysine was without significant effects in both periods. Omission of calcium ions during the postlabelling period inhibited the release of both immunoreactive and radioactive hormone. 2) When amino acids or sulphonylureas were added after prelabelling no signifcant changes were found in the specific radioactivity of released (pro-)insulin or in the fractional release of newly synthesized hormone. Enhanced release of fresh granules from the beta cell might explain the increased fractional release of newly synthesized (pro-) insulin when labelling is carried out in the presence of arginine and sulphonylureas, especially glibenclamide.