血清CRP, PCT, 脂肪酶与胰腺炎分型及预后的相关性研究

目的:探讨血清CRP、PCT和脂肪酶和胰腺炎分型及预后的相关性。方法:87例急性胰腺炎患者,其中轻型急性胰腺炎65例,重症急性胰腺炎22例。在入院后1 d、3 d、7 d、14 d测定血清CRP、PCT、脂肪酶水平并进行APACHEⅡ评分。对比分析两组患者间不同时间点CRP、PCT、脂肪酶及APACHEⅡ评分差异,采用多元线性回归分析影响胰腺炎预后的影响因素。结果:胰腺炎发病严重程度和糖尿病、饮酒史、胆源性结石病有一定的相关性(P0.05)。轻型组CRP、PCT水平1 d、3 d水平和重型组无统计学差异(p0.05);7 d及14 d差异有统计学意义(P0.05)。轻型组脂肪酶水平和重型组3 d、7 d及14 d差异有统计学意义(P0.05)。轻型组APACHEⅡ评分和重型组相比,在发病后1 d、3 d、7 d及14 d差异有统计学意义(p0.05)。回归方程:Logit P=0.0017×PCT+0.0297×胆源性结石病史+0.093×APACHEⅡ评分-0.193。结论:糖尿病和胆源性结石和重症胰腺炎相关,PCT、APACHEⅡ持续不下降提示胰腺炎预后不佳。     

Fluorescent diphenylphosphonate-based probes for detection of serine protease activity during inflammation

Activity-based probes are small molecules that covalently bind to the active site of a protease in an activity-dependent manner. We synthesized and characterized two fluorescent activity-based probes that target serine proteases with trypsin-like or elastase-like activity. We assessed the selectivity and potency of these probes against recombinant enzymes and demonstrated that while they are efficacious at labeling active proteases in complex protein mixtures in vitro, they are less valuable for in vivo studies. We used these probes to evaluate serine protease activity in two mouse models of acute inflammation, including pancreatitis and colitis. As anticipated, the activity of trypsin-like proteases was increased during pancreatitis. Levels of elastase-like proteases were low in pancreatic lysates and colonic luminal fluids, whether healthy or inflamed. Exogenously added recombinant neutrophil elastase was inhibited upon incubation with these samples, an effect that was augmented in inflamed samples compared to controls. These data suggest that endogenous inhibitors and elastase-degrading proteases are upregulated during inflammation.     

Phenolic compounds isolated from Dioscorea zingiberensis protect against pancreatic acinar cells necrosis induced by sodium taurocholate

One new bibenzyl (1) and one new phenanthrene (2), together with two known bibenzyls (3–4) and four known diarylheptanoids (5–8) were isolated from the rhizomes of Dioscorea zingiberensis. The structures of 1–2 were elucidated by spectroscopic methods including 1D and 2D NMR. Phenols 1–8 were evaluated for their anti-pancreatitic activities on sodium taurocholate (NaT)-induced pancreatic acinars necrosis. Notably, 0.5 mM of compound 6 exhibited comparable inhibitory effect with 5 mM of caffeine. Furthermore, compound 6 prevented the ATP depletion and excessive ROS production which could be also involved in mitochondria-mediated injuries in acute pancreatitis. As a result, compound 6 has been demonstrated to be a potential candidate for mediating mitochondrial dysfunction to prevent pancreatic necrosis. This study is also the first report on the isolation of bibenzyls and diarylheptanoids from this plant.     

急性胰腺炎患者腹腔镜胆囊切除术的时机研究

目的：探讨急性胰腺炎胆囊切除术的有效性及时机。方法：分别对患有急性胰腺炎而进行腹腔镜胆囊切除的38例患者的临床信息进行了研究。结果：首先对38例病人进行了3-15天非手术的保守治疗,当临床症状和表征基本消失,血、尿中的淀粉酶接近正常水平时,进行胆囊切除。手术时间为30分钟到90分钟（平均时间60分钟）,手术中无转化和严重并发症症状发生。对这38例病人随访6到56周后无复发现象发生。结论：当临床症状和表征基本消失,血、尿中的淀粉酶接近正常水平时,急性胰腺炎胆囊切除术是有效且可行的。在中国患有急性胰腺炎的患者中,进行腹腔镜胆囊切除的占50％～70％。胆结石治疗取决于患者胰腺炎的严重性,及患者是否患有梗阻性黄疸。对于非梗阻性胰腺炎的患者,早期是否要进行明确的手术治疗尚无定论。我院患有急性胰腺炎的38例患者进行腹腔镜胆囊切除术（2004年2月到2009年4月之间）后取得了满意的效果。详情如下。     

急性胰腺炎1796例病因及治疗效果分析

目的：分析我院急性胰腺炎（AP）的主要病因及治疗效果,为临床诊疗工作提供参考。方法：回顾性分析2008年1月至2012年12月于我院住院治疗的1796例AP患者的临床资料,统计其病因及治疗效果,对其病因及预后进行分析总结。结果：在1796例患者中,重症急性胰腺炎（SAP）522例,轻症急性胰腺炎（MAP）1274例。胆源性AP1025例,高脂血症性AP209例,酒精性AP200例。高脂血疰性AP中,SAP占49．8％,高于胆道疾病及酒精因素引起的SAP的比例（P〈0．05）。33例患者死亡,总病死率1．8％。结论：胆道疾病、酒精、高脂血症是我国AP主要病因,随着医疗水平的提高,AP的病死率有所下降。     

Preparation of Pancreatic Acinar Cells for the Purpose of Calcium Imaging,Cell Injury Measurements,and Adenoviral Infection

The pancreatic acinar cell is the main parenchymal cell of the exocrine pancreas and plays a primary role in the secretion of pancreatic enzymes into the pancreatic duct. It is also the site for the initiation of pancreatitis. Here we describe how acinar cells are isolated from whole pancreas tissue and intracellular calcium signals are measured. In addition, we describe the techniques of transfecting these cells with adenoviral constructs, and subsequently measuring the leakage of lactate dehydrogenase, a marker of cell injury, during conditions that induce acinar cell injury in vitro. These techniques provide a powerful tool to characterize acinar cell physiology and pathology.     

The secretagogue effect of the poison from Centruroides limpidus limpidus on the pancreas of mice and the antagonistic action of the Bouvardia ternifolia extract

In Mexican traditional medicine the plant species Bouvardia ternifolia is used as remedy to treat patients who have been stung by scorpions. In the preceding study, the methanol extract from the roots of this plant was capable of reducing the poisonous effect of Centruroides limpidus limpidus on mice. The poisoning from scorpion C. limpidus limpidus includes manifestations associated with the pancreatitis. This study evaluated the effect produced by the hexane and methanol extract from the root of B. ternifolia upon the acutely inflamed pancreas induced by the venom of C. limpidus limpidus on rats, and the release of amylase in the isolated pancreas of mice. The intravenous administration of venom induced the extravasation of labelled albumin, in a dose dependant manner. The pre-administration of both extracts of Bouvardia ternifolia reduced significantly (p < 0.05) the extravasation by 60%. Upon measuring the secretagogue effect of the venom in the isolated pancreas of mice, the EC50 of the venom was 3.76 x 10(-3) mg ml(-1), whilst in the presence of the methanol and hexane extracts, this EC50 was 9.13 x 10(-3) mg ml(-1) and 0.01629 mg ml(-1). In conclusion, the C. limpidus limpidus venom possesses a secretagogue effect of amylase on the pancreas of mice and produces an inflamed pancreas which is effectively antagonised by the hexane and methanol extracts from the roots of B. ternifolia.     

Autoactivation of Mouse Trypsinogens Is Regulated by Chymotrypsin C via Cleavage of the Autolysis Loop

Chymotrypsin C (CTRC) is a proteolytic regulator of trypsinogen autoactivation in humans. CTRC cleavage of the trypsinogen activation peptide stimulates autoactivation, whereas cleavage of the calcium binding loop promotes trypsinogen degradation. Trypsinogen mutations that alter these regulatory cleavages lead to increased intrapancreatic trypsinogen activation and cause hereditary pancreatitis. The aim of this study was to characterize the regulation of autoactivation of mouse trypsinogens by mouse Ctrc. We found that the mouse pancreas expresses four trypsinogen isoforms to high levels, T7, T8, T9, and T20. Only the T7 activation peptide was cleaved by mouse Ctrc, causing negligible stimulation of autoactivation. Surprisingly, mouse Ctrc poorly cleaved the calcium binding loop in all mouse trypsinogens. In contrast, mouse Ctrc readily cleaved the Phe-150–Gly-151 peptide bond in the autolysis loop of T8 and T9 and inhibited autoactivation. Mouse chymotrypsin B also cleaved the same peptide bond but was 7-fold slower. T7 was less sensitive to chymotryptic regulation, which involved slow cleavage of the Leu-149–Ser-150 peptide bond in the autolysis loop. Modeling indicated steric proximity of the autolysis loop and the activation peptide in trypsinogen, suggesting the cleaved autolysis loop may directly interfere with activation. We conclude that autoactivation of mouse trypsinogens is under the control of mouse Ctrc with some notable differences from the human situation. Thus, cleavage of the trypsinogen activation peptide or the calcium binding loop by Ctrc is unimportant. Instead, inhibition of autoactivation via cleavage of the autolysis loop is the dominant mechanism that can mitigate intrapancreatic trypsinogen activation.     

哺乳期BALB/c小鼠胆道梗阻模型的探索

目的探讨新生BALB/c小鼠胆道梗阻模型的建立,并与报告的新生BALB/c小鼠感染猕猴轮状病毒(RRV)模型小鼠生存曲线进行比较。方法将出生后5～7 d的BALB/c小鼠随机分为实验组和对照组,实验组进行胆总管结扎,然后关腹。对照组打开腹部后关腹不结扎胆总管。实验完成后每天观察小鼠的体重变化、无毛区皮肤颜色变化、小鼠存活天数以及在术后第5、10天时分别取小鼠肝脏做病理及免疫组化。结果小鼠在结扎后随着时间的延长,小鼠的体重及肝体比、无毛区皮肤颜色、存活天数、肝脏病理等都存在一定变化。小鼠体重增长逐渐缓慢,术后第2天就会出现无毛区的皮肤变黄,在尿道口会有淡黄色的液体并随后出现陶土样便。在术后第5天及第10天时取肝脏做肝体比有统计学差异(P≤0.05),小鼠在术后第10天左右会出现死亡高峰。结论新生BALB/c小鼠胆总管结扎模型是研究胆道梗阻的可靠动物实验,其生存曲线与报告的猕猴轮状病毒致胆道闭锁大体类似。