急性坏死性胰腺炎早期肠源性感染的实验研究

目的 通过PUC18质粒载体菌（JM109）示踪法，观察急性坏死性胰腺炎（ANP）时肠屏障损伤与肠道细菌移位情况，探讨ANP继发感染的机理。方法 杂种犬15只于定植PUC18质粒菌JM109后，分ANP组（n=8)和对照组（n=7)。采用气相色谱法测定尿中甘露醇和乳果糖的含量，测定血中二胺氧化酶（DAO）活性和内毒素（LPS）浓度；每日血培养；第7天活杀后，各组脏器标本作细菌培养，肠粘膜菌群分析，所有而氨苄青霉素菌作质粒DNA分析结果 ANP犬肠粘膜上皮绒毛破损，肠通透性显著增加，血浆DAO活性下降，肠道微生态出现明显紊乱，血和脏器出现细菌移位，血中LPS浓度升高1-3倍，ANP犬胰腺中PUC18质粒菌移位率为87.5%。结论 本研究提示，ANP后肠屏障功能严重受损，发生肠道细菌移位。肠源性感染是ANP继发感染的根源。     

急性坏死性胰腺炎后肠道细菌与胰腺感染关系的研究

为探讨急性坏死性胰腺炎的感染源，将ＰＵＣ１８质粒菌ＪＭ１０９置入１５只犬于肠道内定植后，随机分为胰腺炎组及对照组。每日抽血作细菌培养，第７天活杀后，作脏器细菌培养。对耐氮苄青霉素菌株进行酶切分析及琼脂糖凝胶电泳。结果显示，除肾及脾脏外，其余脏器均发现对ＪＭ１０９菌移位，以胰腺移位率最高；血培养及腹水中同样找到质粒菌ＪＭ１０９，发病后２４ｈ血培养阳性率高达４／８。     

急性坏死性胰腺炎后肠道微生态失调与细菌移位关系的研究

15只杂种犬，分为实验组（n＝8）和对照组（n＝7）。七天后活杀。结果发现，实验组犬肠粘膜及内容物中大肠杆菌计数高出对照组10－300倍，而双歧杆菌及乳杆菌则明显减少（P＜0.01）；肠粘膜双歧杆菌／大肠杆菌比值严重倒置（P＜0．05）；肝、胰、脾、肺、肾及肠系膜淋巴结出现了肠道细菌移位；第1、2天血培养结果分别为6／8和5／8。本研究证实，急性坏死性胰腺炎后肠道出现明显的微生态失调，肠道细菌移位到胰腺及其它脏器，成为胰腺及胰周感染的根源。     

急性坏死性胰腺炎早期肠源性感染的实验研究

目的通过ＰＵＣ１８质粒载体菌（ＪＭ１０９）示踪法,观察急性坏死性胰腺炎（ＡＮＰ）时肠屏障损伤与肠道细菌移位情况,探讨ＡＮＰ继发感染的机理。方法杂种犬１５只于定植ＰＵＣ１８质粒菌ＪＭ１０９后,分ＡＮＰ组（ｎ＝８）和对照组（ｎ＝７）。采用气相色谱法测定尿中甘露醇和乳果糖的含量：测定血中二胺氧化酶（ＤＡＯ）活性和内毒素（ＬＰＳ）浓度：每日血培养;第７天活杀后,各组脏器标本作细菌培养,肠粘膜菌群分析,所有耐氨苄青霉素菌作质粒ＤＮＡ分析。结果ＡＮＰ犬肠粘膜上皮绒毛破损,肠通透性显著增加,血浆ＤＡＯ活性下降,肠道微生态出现明显紊乱,血和脏器出现细菌移位,血中ＬＰＳ浓度升高１－３倍,ＡＮＰ犬胰腺中ＰＵＣ１８质粒菌移位率为８７．５％。结论本研究提示,ＡＮＰ后肠屏障功能严重受损,发生肠道细菌移位,肠源性感染是ＡＮ－Ｐ继发感染的根源。     

大黄治疗坏死性胰腺炎的机理探讨

为探讨大黄治疗坏死性胰腺炎的效果和机理，作者采用脱氧胆酸胰管注入法制备成大鼠坏死性胰腺炎模型，术后每８小时分别经口饲喂１０％的大黄汤剂１．５ｍｌ（治疗组，ｎ＝８）或生理盐水１．５ｍｌ（对照组，ｎ＝９）。模拟手术组（模拟组，ｎ＝８）大鼠仅胰管注射生理盐水，作为对照。术后４８小时处死动物。分别测定肠运动，血内毒素和淀粉酶水平，并做肠系膜淋巴结和胰腺的细菌培养。结果：对照组中５只大鼠（５／９），治疗组中１只大鼠（１／８）在实验期内死亡，模拟组无死亡；对照组较模拟组有显著的肠运动抑制，而治疗组较对照组肠运动明显改善；对照组大鼠的血内毒素（６１．３６±２８．３０ｐｇ／Ｌ）明显高于治疗组（５．４１±３．５８ｐｇ／Ｌ，Ｐ＜０．００１）；模拟组大鼠无明显的内毒素血症；对照组的淋巴结（４／４）和胰腺（４／４）细菌培养均阳性，而治疗组中淋巴结和胰腺仅各１份（１／７）培养阳性，模拟组的淋巴结和胰腺培养均阴性。以上结果说明大黄通过促进肠运动，能防治坏死性胰腺炎时的细菌移位，避免胰腺感染，改善坏死性胰腺炎的预后。     

大黄对重症急性胰腺炎的影响及细菌移位研究

目的：采用牛磺胆酸钠诱发大鼠重症急性胰腺炎模型,探讨大黄对重症急性胰腺炎的影响及细菌移位。方法：健康Wistar大鼠30只,雌雄不限,随机分成三组。1组为SAP模型组（M组,n=10）,开腹后肝端胰胆管用小动脉夹夹闭,肠端胰胆管用丝线结扎。5%牛磺胆酸钠溶液0.1ml/100g胰胆管内以0.2ml/min速度逆行推注,复制SAP模型; 2组为中药大黄治疗组（T组,n=10）,按M组方法复制SAP模型,并在复制模型后即刻胃管内注入生大黄粉溶液1ml（5mg/100g）。3组为假手术组（SO组,n=10）,开腹后自胰管内逆行注入生理盐水0.1ml/100g后关腹。术后取血液、胰腺和肠系膜淋巴结进行细菌培养。结果：脏器及血细菌培养结果显示,SO组血培养阴性,除2只大鼠肠系膜淋巴结培养出细菌外,其他脏器培养均阴性。M组细菌培养阳性率较高（P〈0.01）,T组细菌培养阳性率明显低于M组（P〈0.01）。结论：SAP时细菌移位的发生率较高,可能与机体的细胞免疫功能下降有关。大黄能抑制肠菌及清理肠道,从而降低胰腺感染率,对SAP有一定的治疗作用。     

CO_2气腹对腹膜炎大鼠的肠道内毒素细菌移位的影响

目的 探讨不同程度CO2气腹对腹膜炎大鼠肠道内毒素细菌移位的影响.方法 采用PUCl8质粒载体大肠杆菌(JM109)示踪法,将质粒菌JM109定植于大鼠肠道后,腹腔内注入盐酸建立无菌的急性腹膜炎大鼠模型,48只大鼠随机均分为3组,分别为对照组、5 mmHg CO2气腹组、15 mmHg CO2气腹组.每间隔1 h抽股静脉血测内毒素含量;气腹3 h后,取门静脉血、肠系膜淋巴结作细菌培养,对耐氨苄青霉素菌株进行酶切分析及琼脂糖凝胶电泳;并测回肠黏膜及门静脉血中分泌型免疫球蛋白A含量.结果 气腹1 h后,15 mmHg CO2气腹组股静脉血内毒素含量开始显著升高(P<0.01),并随气腹时间延长而进行性升高;气腹3 h后,15 mmHg CO2气腹组门静脉血和肠黏膜SIgA含量明显低于对照组(P<0.01),肠道大肠杆菌(JM109)至门静脉血的移位率分别为0、25%、5O%,至肠系膜淋巴结的移位率分别为12.5%、25%、75%.结论 CO2气腹能够促进腹膜炎大鼠的肠道内毒素细菌移位,CO2气腹压为15 mmHg时作用更明显.     

急性胰腺炎病人免疫功能的变化与细菌移位及大黄干预的关系

目的探讨重型急性胰腺炎（severeacutepancreatitis，SAP）病人免疫功能的变化与细菌移位及大黄干预的关系。方法健康Wistar大鼠30只，随机分成三组。SAP模型组（M组，n=10）：5％牛磺胆酸钠溶液0．1ml·100g^-1胰胆管内以0．2ml／min速度逆行推注，复制模型；大黄治疗组（T组，n=10）：在复制模型后即刻胃管内注入精制大黄粉溶液1m1（5mg·100g^-1）；假手术组（S0组，n=10）：开腹后自胰管内逆行注入生理盐水0．1ml·100g^-1后关腹。24h后用流式细胞仪检测SAP大鼠T淋巴细胞亚群CD4＋、CD8＋阳性细胞百分比并计算CD4＋／CD8＋的值；对血液、胰腺和肠系膜淋巴结进行细菌培养。结果M组血中CD4阳性细胞数以及CD4＋／CD8＋值明显低于S0组及T组（P〈0．01，P〈0．05）。M组细菌培养阳性率较高，T组细菌培养阳性率明显低于M组（P〈0．01）。结论SAP时存在细胞免疫功能紊乱，CIM阳性细胞数以及CD4＋／CD8＋值明显降低。SAP后肠细菌移位与机体的细胞免疫功能紊乱有关。大黄能上调SAP血中CD4阳性细胞数以及CD4＋／CD8＋值，对SAP有一定治疗作用。     

生长抑素、宫入菌干预急性水肿性胰腺炎的病理损伤影响

目的探讨生长抑素、宫入菌培菲康干预急性水肿性胰腺炎后对胰腺（AEP）及其远距离器官肺、小肠病理形态学改变的影响。方法SD幼龄大鼠随机分为五组：正常对照组、急性水肿性胰腺炎模型组、生长抑素干预组、培菲康干预组及联合生长抑素、培菲康干预组，AEP幼龄大鼠模型制备采用皮下注射蛙皮素溶液的方法，分别予生长抑素和／或宫入菌干预8h后。观察不同组胰腺、肺及小肠的病理评分变化。结果与正常对照组相比，干预组及模型组胰腺、肺及小肠病理评分职显升高（P均〈0．05）；与模型组相比，各干预组各组织病理评分明显下降（P均〈0．05），生长抑索与宫入菌具交互作用，但联合使用生长抑索和培菲康干预组与单用生长抑索或宫入菌干预组病理评分差异无显著统计意义（P〉0．05）。结论生长抑素和／或培菲康干预急性胰腺炎具减轻胰腺及其远距离脏器如肺或小肠组织的炎症反应的作用，其疗效无显著差异；生长抑素与宫入菌具交互作用。     

急性坏死性胰腺炎后肠道微生态失调与细菌移位关系的研究

１５只杂种犬，分为实验组和对照组，七天后活杀，结果发现，实验组犬肠粘膜及内容物中大肠杆菌计数高出对照组１０－３００倍，而以歧杆菌及乳杆菌及乳杆菌则明显减少；肠粘膜双歧杆菌／大肠杆菌比值严重倒置；肝、胰、脾、肺、肾及肠系膜淋巴结了肠道细菌移位。第１、２天血培养结果分别为６／８和５／８，本证实，急性坏死性胰腺炎后肠道出现明显的微生态失调，肠道细菌称位到胰腺及其它脏器，成为胰腺及胰周感染的根源。     

急性胰腺炎肠黏膜损伤情况及通透性改变的实验研究

目的：观察实验性重症急性胰腺炎（ANP）鼠肠通透性的改变,并观察肠黏膜形态变化及细菌移位的关系。方法：实验性大鼠41只,随机分为胰腺炎组和对照组。利用电子显微镜以硝酸镧作为踪剂观察肠黏膜通透性和肠细胞的渗透性。结果：回肠黏膜出现明显病理改变,肠黏膜通透性和肠细胞渗透性明显增高。胰腺炎组肠系膜淋巴结和腹水标本培养出肠道菌属者,结论：ANP时存在着严重的肠道屏障功能损伤。发病时肠黏膜即遭到严重破坏,肠通透性增高,从而引起肠道细菌移位。     

恶性疟原虫红内期SSUrRNA编码基因的克隆与序列分析

目的 克隆恶性疟原虫海南株红内期小亚单位核糖体核糖核酸SSUrRNA编码基因(SSUrDNA)片段，并进行序列分析。方法根据献报道恶性疟原虫基因序列设计1对特异性引物，采用聚合酶链反应(PCR)方法从海南恶性疟患血样核酸提取物中扩增出恶性疟原虫SSUrDNA目的基因片段，纯化后与PUC19m-T质粒载体连接构建重组子，并导入大肠杆茵JM109；阳性克隆经双酶切鉴定后，双脱氧末端终止法测定序列，并采用ExPASy Proteomicstools软件分析。结果 恶性疟原虫SSUrDNA扩增片段大小约为431bp；阳性克隆重组质粒作双酶切及PCR扩增均得到预期大小的基因片段；测定的SSUrDNA插入片段核酸序列与巴西恶性疟原虫IMTM／7G8相同基因序列对比。未发现碱基的插入或缺失去，同源性为99．3％；第353位碱基由C取代了G，第371位碱基由T取代了C，其余序列相同。结论 成功克隆了恶性疟原虫海南株红内期SSUrDNA基因片段，该序列在不同恶性疟原虫虫株间相对保守。     

大鼠急性坏死性胰腺炎肠黏膜屏障功能障碍的观察

目的: 观察急性坏死性胰腺炎(ANP)大鼠肠黏膜屏障功能的变化。方法: SD大鼠80只随机分为假手术组(n=40)和ANP组(n=40)。胆胰管内逆行注射5%牛磺胆酸钠溶液制作ANP模型。观察大鼠胰腺和回肠的病理变化,动态测定肠脂肪酸结合蛋白(IFABP)、内毒素水平及肠系膜淋巴结和门静脉血细菌移位率。结果: 模型制作后2h血清IFABP明显升高,6h达到峰值(P<0.01),24h后降低。早期内毒素水平有明显升高,48h达到峰值(P<0.01)。肠系膜淋巴结细菌移位在ANP24h后明显升高,48h达到6/8只(P=0.013),门静脉血细菌移位48h达到3/8只(P=0.216)。结论: ANP早期肠黏膜屏障功能受损,引发肠道细菌和内毒素移位,IFABP可能是ANP早期肠黏膜屏障功能受损的预警指标。     

急性坏死性胰腺炎肠通透性及病理改变实验观察

目的观察急性坏死性胰腺炎（ＡＮＰ）犬肠通透性变化及肠粘膜病理改变。方法１５只杂种犬,分为ＡＮＰ组（ｎ＝８）和对照组（ｎ＝７）,经主胰管注入牛磺胆酸钠和胰蛋白酶复制ＡＮＰ模型。用气相色谱法检测尿中乳果糖／甘露醇（Ｌ／Ｍ）比值,第７天剖杀,取回肠粘膜作病理观察。结果ＡＮＰ犬尿中Ｌ／Ｍ比值高出对照组２～１０倍。肠粘膜顶端上皮脱落,脱落处有大量细菌侵入,微绒毛部分脱落,面积减少。血浆内毒素水平高出对照组１～２倍,二胺氧化酶活性下降,细菌移位率１００％。结论ＡＮＰ时肠粘膜严重受损,监测尿中Ｌ／Ｍ比值变化可反映早期肠粘膜损伤。     

基因重组碱性成纤维细胞生长因子对急性坏死性胰腺炎犬肠道细菌移位的影响

目的探讨基因重组碱性成纤维细胞生长因子（ｂＦＧＦ）对急性坏死性胰腺炎（ＡＮＰ）犬肠道细菌移位的影响。方法杂种犬２３只,分对照组（ｎ＝７）、ＡＮＰ组（ｎ＝８）和ｂＦＧＦ组（ｎ＝８）。ｂＦＧＦ组犬复制ＡＮＰ模型后,每日静脉注射ｂＦＧＦ（５μｇ／ｋｇ）。结果ｂＦＧＦ治疗后,ＡＮＰ犬肠粘膜损伤明显减轻,脏器细菌培养阳性率下降５０％,细菌移位数量减少９３．３％－９６．７％,肠粘膜蛋白质、ＤＮＡ含量显著增加（Ｐ＜０．０５）,丙二醛含量明显减少（Ｐ＜０．０５）。结论ｂＦＧＦ可显著减少ＡＮＰ时肠道细菌移位,其机制可能是通过增加肠粘膜蛋白质合成,促进肠粘膜损伤修复。     

Effects of severe acute pancreatitis on gut bacteria/endotoxin translocation in pigs

Objective： To set up a swine model of severe acute pancreatitis（SAP） and to observe its relationship with the gut-originated bacteria/endotoxin translocation. Methods： Forty pigs weighing 17-22 kg were randomly diyided into SAP group （n=34） and sham-SAP group （n=6）. By injecting 1 ml/kg of combined solution of 5% sodium taurocholate and 8 000-10 000 benzoyl arginine ethyl ester（BAEE） units trypsin per milliliter into pancreas via pancreatic duct, SAP was induced under anesthesia. Endotoxin samples from vena cava were determined by chromogenic limulus amebocyte lysate （LAL） technique. Both portal and central vena blood samples were collected before and 72 h after the induction of SAP and cultured for both aerobic and anaerobic bacterial growth. Animals were sacrificed at the end of experiments by injecting 20 ml of 10% KCl intravenously and tissue specimens of mesenteriolum and mesocolon lymph nodes, lung, pulmonary portal lymph nods and pancreas were taken immediately after animal death, and homogenized for bacteriological studies. Results： Systemic plasma endotoxin levels increased rapidly 6 h post induction of SAP（PIS） with a peak at 48 h PIS （P〈0.01）. The magnitude of bacterial translocation in both portal and systemic blood and remote systemic organs as well were recovered PIS. Conclusion： （1） A swine model of SAP was established; （2）The early endotoxemia PIS seamed probable originated from gut endotoxin translocation; （3）The magnitude of bacterial translocation in both portal and systemic blood and the remote systemic organs as well were recovered at 72h PIS.     

Obstructed Bile Duct as a Trigger for Microbe‘s Translocation？

To assess the potential mechanisms of bacterial translocation in a murine model of obstructive jaundice. Methods: Adult rats were randomized to be operated on for ligation or sham-ligation of the common bile duct. Bacterial translocation to the mesenteric lymph nodes (MLNs), liver, spleen, portal blood and systemic circulation and bacterial population levels in the ceca were quantitated after 7 and 14 days. The terminal ilea were histologically examined by light and transmission electron microscopy. Results: Bacterial translocation to the MNLs was seen in both 7 (10/17) and 14 (11/18) day ligated animals, but not found in their corresponding controls (both 0/8). No significant difference in the cecal bacterial population levels was found between the ligated groups and their corresponding control groups, also between the two subgroups that were set up within each ligated group according to the presence or absence of bacteria in the MLNs. In the ligated rats, light microscopy demonstrated subepithelial edema in association with infiltration of flammatory cells and, transmission electron microscopy showed that the enterocytes were injured with abnormal microvilli, swollen mitochondria, unclear endoplasmic reticulum and cytoplasm with bubble degeneration. However, the ilea from the controls appeared normal. Conclusions: Obstructive jaundice promotes bacterial translocation in rats. The gut mucosal damage rather than the intestinal bacterial overgrowth may play a crucial role in bacterial translocation.     

Obstructed Bile Duct as a Trigger for Microbe's Translocation?

To assess the potential mechanisms of bacterial translocation in a murine model of obstructive jaundice. Methods: Adult rats were randomized to be operated on for ligation or sham-ligation of the common bile duct. Bacterial translocation to the mesenteric lymph nodes (MLNs), liver, spleen, portal blood and systemic circulation and bacterial population levels in the ceca were quantitated after 7 and 14 days. The terminal ilea were histologically examined by light and transmission electron microscopy. Results: Bacterial translocation to the MNLs was seen in both 7 (10/17) and 14 (11/18) day ligated animals, but not found in their corresponding controls (both 0/8). No significant difference in the cecal bacterial population levels was found between the ligated groups and their corresponding control groups, also between the two subgroups that were set up within each ligated group according to the presence or absence of bacteria in the MLNs. In the ligated rats, light microscopy demonstrated subepithelial edema in association with infiltration of flammatory cells and, transmission electron microscopy showed that the enterocytes were injured with abnormal microvilli, swollen mitochondria, unclear endoplasmic reticulum and cytoplasm with bubble degeneration. However, the ilea from the controls appeared normal. Conclusions: Obstructive jaundice promotes bacterial translocation in rats. The gut mucosal damage rather than the intestinal bacterial overgrowth may play a crucial role in bacterial translocation.