健脾疏肝方对反流性食管炎大鼠模型食管组织病理及PCNA的影响

目的:观察健脾疏肝方对反流性食管炎大鼠模型食管组织病理及增殖细胞核抗原(PCNA)表达的影响.方法:采用"不全幽门结扎 贲门肌切开术"制作动物模型,240只大鼠分为正常组、模型组、安慰剂(谷维素)组、阳性对照组(加斯清)、治疗高、中、低剂量组.给予相应处理后,取食管组织行HE染色,光镜下观察组织病理学变化,用免疫组化的方法测PCNA表达.结果:健脾疏肝方能显著改变反流性食管炎的病理变化,健脾疏肝方高剂量组食道组织病理变化明显减轻;健脾疏肝方能显著抑制PCNA表达的升高,与模型组相比PCNA的表达显著降低(小剂量:284.83±30.49vs 330.00±35-35,P＜0.05:中剂量:239.67±28.84vs330.00±35-35,P＜0.01:高剂量:203.00±25.29vs330.00±35.35,P＜0.01),其疗效与加斯清相当.结论:健脾疏肝方有助于降低PCNA的高表达,并有抗胃食管反流,减轻食管炎症的疗效.     

健胃冲剂对反流性食管炎模型大鼠血清胃泌素和胃动素分泌的影响

[目的]探索健胃冲剂治疗反流性食管炎(RE)的作用机制.[方法]采用“不全幽门结扎十贲门肌切开术”制作RE大鼠模型,将术后恢复较好的大鼠随机分为5组,即阳性对照组,西药对照组,中药高、中、低3个剂量组,同时设正常组,术后3周开始灌胃给药,连续给药3周后,腹主动脉取血,应用ELISA法检测各组大鼠血清胃泌素(GAS)和胃动素(MTL)的含量.[结果]中药高、中、低剂量组及西药对照组与阳性对照组相比,血清GAS及MTL含量均增高(P＜0.05),中药低剂量组对于提高MTL含量优于西药对照组(P＜0.05).[结论]健胃冲剂有助于调节GAS及MTL的分泌,使胃肠道运动功能增强,促进胃肠排空,减少胃内容物反流,也可提高食管下括约肌紧张性,增强廓清功能,从而对RE起到治疗作用.     

一氧化氮在实验性反流性食管炎发病机制中的作用

目的　探讨一氧化氮(NO)在实验性反流性食管炎发病中的作用。方法　48 只SD大鼠随机分为3 组:A组:采用完全幽门结扎 贲门肌切开术;B组:采用半幽门缝扎 贲门肌切开术,分别制备反流性食管炎动物模型;C组:假手术对照组。并于模型制备后24小时、48 小时、72 小时观察食管下端粘膜的病理表现,测定食管组织的NO含量。结果　食管炎指数(肉眼和显微镜下),A 组术后24小时以及B组术后24 小时、48 小时、72 小时与C组比较差异均有非常显著意义( P< 0.01 );食管组织NO含量,A 组术后24小时以及B组术后24小时、48小时、72小时与C组比较,差异亦均有显著性意义(P< 0.05, P< 0.01 );不同程度食管炎组的食管组织NO 含量均明显高于非食管炎组(P<0.05,P< 0.01 ),而不同程度食管炎组间的食管组织NO含量差异无显著性意义(P> 0.05 )。结论　NO在实验性反流性食管炎发病机制中对食管粘膜可能发挥损伤和保护的双重作用     

环形肌切开和全层肌切开治疗贲门失弛缓症长期随访研究

目的：对比内镜下全层肌切开与环形肌切开治疗贲门失弛缓症的长期临床疗效及远期并发症。方法：回顾性分析2012年6月至2014年12月于郑州大学第一附属医院消化内科行经口内镜下肌切开术治疗并定期随访的53例贲门失弛缓症患者资料,其中21例行环形肌切开,32例行全层肌切开,比较两种术式的长期临床疗效及远期并发症。 结果：环形肌切开组和全层肌切开组治疗有效率分别为90.5%（19/21）和100%（32/32）。两组术后Eckardt评分、食管下括约肌压力和4 s完整松弛压比较差异无统计学意义（P>0.05）。全层肌切开组临床相关胃食管反流发生率高于环形肌切开组（40.6％比14.3%,χ2=4.174,P=0.041）。 结论：经口内镜下环形肌切开术与全层肌切开术治疗贲门失弛缓症长期疗效相当,但全层肌切开术后临床相关胃食管反流发生率更高。     

旋覆代赭汤对酸性反流性食管炎模型大鼠食管下段粘膜pH值的影响

目的 :观察旋覆代赭汤对酸性反流性食管炎模型大鼠食管下段粘膜 p H值的影响。方法 :用不全幽门结扎加食管下括约肌 (L ES)切开术制备酸性反流性食管炎大鼠模型后 ,给予旋覆代赭汤水煎剂 ,分别于治疗 2、4、8d后观察各组大鼠食管下段粘膜 p H值的变化。结果 :用旋覆代赭汤治疗 4 d后 ,食管下段粘膜 p H值即有明显改善 ,与假手术对照组相比差异无显著性意义 (P >0 .0 5 )。结论 :旋覆代赭汤在治疗酸性反流性食管炎的过程中 ,通过抑制酸反流、抗炎等作用 ,明显提高食管下段粘膜的 p H值 ,从而促进疾病的恢复。     

冠脉结扎法大鼠急性心肌梗死的造模

目的 探讨冠脉结扎法制作大鼠急性心肌梗死模型。 方法 40只Sprauge-Dawley大鼠随机分为冠脉结扎组和假手术组,大鼠经麻醉后,气管插管连接小动物呼吸机,打开左侧胸腔,暴露心脏,结扎左冠状动脉前降支,假手术组只用线穿过左前降支而不结 扎,其余步骤同冠脉结扎组。手术前后均行心电图检查,4周后行血流动力学检测,取出心脏,观察其外形变化,行HE染色后于显微镜下观察其病理变化。 结果 冠脉结扎组制作心梗模型成功率为100%,术后存活率为65%,假手术组术后存活率为80%。冠脉结扎组术后4周取出的心脏,左心室较假手术组扩大,缺血区室壁变薄。 结论 冠脉结扎法制作大鼠心肌梗死模型,效果稳定,成功率高,制作方便,经济易推广。     

不同时点分别结扎左、右颈总动脉建立大鼠血管性痴呆模型

目的 改良大鼠血管性痴呆(VD)双侧颈总动脉永久性结扎模型,提高模型动物存活率.方法 采取间隔3 d分2次结扎双侧颈总动脉建立VD模型,观察大鼠的成活率,及术后4 w和8 w学习记忆能力变化.结果 改良模型组动物存活率(96.0%)明显高于传统模型.术后4 w和8 w,该模型组大鼠学习记忆能力障碍明显,显著低于假手术对照组.结论 间断永久性结扎双侧颈总动脉法是建造大鼠VD模型的理想方法.     

活血通降方对反流性食管炎大鼠食管内脏高敏感及LPS/TLR4/MC通路的影响

目的:研究活血通降方对反流性食管炎(reflux esophagitis, RE)模型大鼠脂多糖(lipopolysaccharide, LPS)/Toll样受体4(Toll-like receptor 4,TLR4)/肥大细胞(mast cell, MC)通路的影响，探讨其对食管内脏高敏感的疗效机制。方法:将50只Wistar大鼠随机分为对照组、模型组、西药组、中药高剂量组及中药低剂量组5个组，每组10只。对照组行腹腔开关术，其余各组均采用改良部分贲门肌切开术+外置幽门部分结扎术联合束缚应激建立RE大鼠模型。造模7 d后，各组分别给予相应剂量药物或相同体积生理盐水灌胃，连续给药14 d, 2次/d。苏木精-伊红染色评价食管黏膜病理表现及评分；采用行为学评分和观察肌电图变化评价大鼠食管的内脏敏感性；采用甲苯胺蓝染色检测食管组织中MC数目；采用免疫组织化学法检测食管组织类胰蛋白酶(tryptase, TPS)和结肠组织中紧密连接蛋白ZO-1及Occludin的表达。采用酶联免疫吸附法检测血清LPS、IL-8含量，采用蛋白免疫印迹法检测食管组织中TLR4的表达，结果:与对照组比较，模型组大...     

活血通降方对反流性食管炎大鼠肠道菌群及Caspase-3/GSDME通路的影响

目的:探讨活血通降方对反流性食管炎大鼠肠道菌群及半胱氨酸蛋白酶3/焦孔素E(Caspase-3/GSDME)通路的影响。方法:将30只Wistar大鼠随机分为对照组、模型组、活血通降方组,每组10只。除对照组外,其余两组均采用改良部分贲门肌切开术+外置幽门部分结扎术建立反流性食管炎大鼠模型。造模成功后,对照组及模型组予2.5 mL生理盐水灌胃,活血通降方组予2.5 mL活血通降方(3.68 g/kg)灌胃,2次/d,连续给药14 d。采用苏木精-伊红染色法观察各组大鼠食管组织病理变化;采用蛋白组学检测差异蛋白变化;采用酶联免疫吸附法检测血清中内毒素、IL-1β和IL-18的表达;采用蛋白免疫印迹法检测食管组织中Caspase-3和GSDME的表达,采用16S rDNA高通量测序检测肠道菌群的变化。结果:模型组大鼠食管组织病理评分较对照组增高(P<0.05),活血通降方组大鼠食管组织病理评分较模型组下降(P<0.05);蛋白组学分析表明,Caspase-3是各组的主要差异蛋白;与对照组比较,模型组肠道菌群的物种丰度显著降低,未分类拟杆菌属、Prevotellaceae＿UCG...     

线栓法建立大鼠局灶性脑缺血模型的改进

目的对常规线栓法制作大鼠脑缺血模型进行改进，以提高模型动物的长期存活率。方法将60只体质量250～270g的SD雄性大鼠随机分成3组，每组20只；对照组采用常规手术方法，术中显露并结扎翼腭动脉，术后自主进食；灌胃组手术方法同对照组，但术后辅助灌胃；改良组不结扎翼腭动脉，但仔细分离颈部动脉与迷走神经，尽量避免操作时牵拉迷走神经，术后辅助灌胃。在模型制作前、后，连续称量各组大鼠的体质量，并行神经功能缺损评分；在造模后3、14、28d，分别计算各组大鼠的存活率；造模后28d，计算各组大鼠的脑梗死体积。结果①改良组模型制作的平均时间为19．5min，低于对照组和灌胃组，差异有统计学意义，P〈0．05；②造模后第3天，改良组大鼠的存活率均高于对照组和灌胃组，差异有统计学意义，P〈0．05；造模后第28天，改良组大鼠的存活率高于对照组，差异有统计学意义，P〈0．05；③造模后3组大鼠的体质量均下降，从造模后第7天开始，各组存活大鼠的体质量均开始回升，在第7、14、21、28天，改良组大鼠的体质量均高于对照组和灌胃组，差异有统计学意义，P〈0．05；④造模后3组大鼠的神经功能缺损评分和脑梗死体积的比较，差异均无统计学意义。结论改进的脑缺血模型制作方法，可缩短制作时间，使大鼠体质量恢复较快，能有效地提高大鼠脑缺血造模后的存活率。     

复方白及糊对反流性食管炎模型大鼠黏膜血流的影响

[目的]探讨复方白及糊治疗反流性食管炎（RE）的作用机制。[方法]实验大鼠随机分为4组,各23只。甲、乙、丙3组均采用改良部分贲门肌切开加外置幽门部分结扎术,制备RE大鼠模型,并与假手术组对照。甲组造模前7天起,每天灌食复方白及糊3ml;造模后与乙组一起每天灌食等量复方白及糊,丙组同时灌等量0.85%氯化钠。分别于术后7、14d观察各组大鼠食管下段pH值、大体标本及组织学改变,同时进行食管黏膜血流量的测定。[结果]造模成功,造模大鼠食管黏膜血流均有所下降,甲、乙组可使模型大鼠食管下段pH升高,损伤黏膜迅速修复,食管下端黏膜血流增加,其中甲组作用更强,与丙组、假手术组比较差异有统计学意义（P〈0.05）。[结论]复方白及糊治疗RE,可能是通过抑酸,改善局部黏膜血供,加强食管黏膜上皮屏障实现的。     

反流性食管炎时食管一氧化氮合酶表达的实验研究

目的：初步探讨反流性食管炎时食管ＮＯＳ的表达。方法：采用幽门半缝扎＋贲门肌切开术制备反流性食管炎动物模型,采用ＮＡＤＰＨ－ｄ组化染色观察食管ＮＯＳ表达,测定食管组织ＮＯ含量。结果：食管炎组大多呈ＮＯＳ强阳性反应,阳性率明显高于正常对照组（Ｐ＜０．００１）,食管组织ＮＯ含量也明显高于正常对照组（术后２４小时比较Ｐ＜０．０１,术后４８小时,７２小时比较均Ｐ＜０．０５）。结论：胃－食管反流可引起食管上皮粘膜ＮＯＳ的过度表达,过多的ＮＯ可能发挥着重要的作用。     

Protective Effect of Ammonia Against Reflux Esophagitis in Rats

Although several recent studies have reported that curing Helicobacter pylori (H. pylori) may result in the development of reflux esophagitis (RE), the mechanisms leading to this complication are unknown. One by product of H. pylori infection is ammonia, which serves as an acid neutralizer. The aim of this study was to clarify whether ammonia, which is produced during H. pylori infection, has a protective effect on the esophagus. Eight-week-old male Sprague-Dawley rats were fasted for 24 hrs. Under anesthesia, both the pylorus and limiting ridge were simultaneously ligated. One hour postligation, 0.3 ml of saline or ammonia at various concentrations was administered intragastrically by gastric intubation. Three hours after ligation, the animals were killed, the esophagus and stomach were removed, and the length of esophageal hemorrhagic erosions was measured. The incidence of RE was 100% (7/7) in the control group, 71% (5/7) in the low-ammonia group, 29% (2/7) in the middle-ammonia group, and 14% (1/7) in the high-ammonia group. The severity of lesions decreased in correspondence to increases in ammonia concentration. The development of RE was significantly inhibited by ammonia in a dose-dependent manner. This study indicates that ammonia protects against development of RE. A decreased amount of ammonia in the stomach might be related to the development of RE after H. pylori eradication therapy.     

Cardiaplasty and Roux-en-Y partial gastrectomy (Serra-Dória procedure) for reoperation of achalasia

BACKGROUND: After cardiomyotomy for the treatment of megaesophagus, recurrence of symptoms occur in up to 15% of the patients, but only some require a reoperation. AIM: To evaluate the results of reoperation -- cardioplasty with Roux-en-Y partial gastrectomy, a technique proposed by Serra-Dória. CASUISTIC AND METHODS: Twenty patients with achalasia previously treated by cardiomyotomy, were retrospectively studied. The etiology of symptoms recurrence was reflux esophagitis in nine (45.0%) patients, healing of the myotomy in five (25.0%), end staging megaesophagus in five (25.0%) and incomplete myotomy in one (5%). Intra and postoperative complications were analyzed. The patients were studied by clinical (dysphagia, regurgitation, heartburn and weight gain), radiological and endoscopic evaluation, in the pre- and postoperative period. RESULTS: Five (25.0%) patients had complications in the immediate postoperative period. No deaths were observed. Dysphagia improved in all the patients. Regurgitation and heartburn almost disappeared in the whole group. Weight was maintained or increased in 64.7% of the patients. Radiological studies showed a decrease in the caliber of the esophagus in 53.0%, while the remaining patients maintained the pre-operative diameter. Endoscopy, performed during the late postoperative period in 17 patients, showed that 6 among the 9 with reflux esophagitis improved; 2 among the 8 with a normal esophagus during the preoperative period, developed esophagitis. CONCLUSIONS: The Serra-Dória procedure for the treatment of megaesophagus in patients who had already undergone cardiomyotomy and whose symptoms recurred, presented a low morbidity and no mortality. It offered a significant relief of symptoms with a decrease of the caliber of the esophagus in several patients. The patients also improved with regards to reflux esophagitis. In some cases reflux was still present after surgery. Others with normal esophagus in the preoperative period developed esophagitis.     

A rat model of esophageal varices

We have developed a new method for inducing portal hypertension and esophageal varices in rats--partial ligation of the portal vein after devascularization of the circumference of the left renal vein and complete ligation of the portal vein on the fifth day thereafter. Thirty rats were separated into groups of 10, control (sham operation), complete portal ligation only and complete portal ligation plus devascularization. Two weeks after the surgery, the presence of esophageal varices in rats with complete portal ligation plus devascularization was confirmed by portography and by the histological findings. The diameter (mean +/- SD) of the submucosal veins of the lower esophagus in the complete portal ligation plus devascularization group (219.4 +/- 86.6 microns) was significantly larger than that in the complete portal ligation group (99.8 +/- 53.4 microns) or in the control group (30.5 +/- 16.6 microns) (p less than 0.01). Vascular structures of the lower esophagus closely resembled those in humans with esophageal varices. This new technique is simple, rapid and reliable, and application can be made to various experimental studies on portal hypertension.     

胃食管反流病与幽门螺杆菌感染的关系探讨

目的探讨胃食管反流病与幽门螺杆菌感染之间的相关性。方法将经过电子胃镜确诊的GERD患者120例及对照组轻度慢性浅表性胃炎患者120例予血清幽门螺杆菌抗体检测和14C呼气试验法进行H.pylori检测,对比两组H.pylori感染情况;将90例反流性食管炎患者分为LA-A、B组及LA-C、D组,对比两组H.pylori感染情况;将120例GERD患者分为轻度症状组、中度症状组、重度症状组及极重度症状组,比较组间H.pylori感染情况。结果 GERD组H.pylori感染的阳性率(39.17%)低于对照组H.pylori感染的阳性率(62.50%),差异有统计学意义(P<0.05)。LA-A、B组H.pylori感染的阳性率(60.87%)高于LA-C、D组H.pylori感染的阳性率(29.55%),差异有统计学意义(P<0.05)。轻度症状组、中度症状组、重度症状组及极重度症状组H.pylori感染的阳性率分别是40.00%、41.67%、40.63%、31.82%。结论幽门螺杆菌感染是反流性食管炎的保护因素,幽门螺杆菌感染与GERD症状的发生无相关性。     

The prevalence of Helicobacter pylori infection and the status of gastric acid secretion in patients with Barrett's esophagus in Japan

OBJECTIVES: The acidity of the refluxate into the esophagus is a key factor for the pathogenesis of gastroesophageal reflux disease. Helicobacter pylori (H. pylori) infection can influence gastric acid secretion. We have reported that H. pylori infection prevents reflux esophagitis by decreasing gastric acid secretion in Japanese patients, but the role of this organism in Barrett's esophagus is unclear. The aim of this study was to investigate the prevalence of H. pylori infection and gastric acid secretion in Japanese patients with reflux esophagitis with or without Barrett's esophagus. METHODS: We enrolled 112 reflux esophagitis patients who were examined for the status of H. pylori and acid secretion in this study. They were divided into three groups, according to the presence or absence of Barrett's esophagus as follows: reflux esophagitis group without Barrett's esophagus (reflux esophagitis alone) (80 patients); short-segment Barrett's esophagus group (16 patients); and long-segment Barrett's esophagus group (LSBE) (16 patients). Age- and sex-matched control subjects were also assigned to the 80 patients with reflux esophagitis alone. The prevalence of H. pylori infection was determined by histology, rapid urease tests, and serum IgG antibodies. Gastric acid secretion was evaluated by the endoscopic gastrin test (EGT). RESULTS: The overall prevalence of H. pylori infection in the reflux esophagitis patient group (24.1%) was significantly lower than the control group (71.2%) (odds ratio 0.13, 95% confidence interval 0.07-0.24; p < 0.0001). The prevalence of H. pylori infection in the patients with Barrett's esophagus tended to be lower than that in the patients with reflux esophagitis alone (reflux esophagitis alone; 30.0%, SSBE; 18.7%, LSBE; 0%), especially in the patients with LSBE compared with the reflux esophagitis alone group (p < 0.01). The EGT value of the respective reflux esophagitis patient group was significantly higher than the control group. The EGT value in the patients with Barrett's esophagus tended to be higher than that in the patients with reflux esophagitis alone, but the difference was not statistically significant. When examined in H. pylori-negative subjects, no difference was found in the EGT value between the control subjects and the patients with reflux esophagitis alone, but it was significantly higher in patients with Barrett's esophagus than the control subjects (p < 0.05). On the other hand, when examined in the H. pylori-positive subjects, the EGT value was significantly higher in the patients with reflux esophagitis alone than in the control subjects (p < 0.01). CONCLUSIONS: H. pylori infection may play a protective role in the development of Barrett's esophagus, especially in the development of LSBE in Japan. Gastric acid hypersecretion may be concerned with the development of Barrett's esophagus in addition to the absence of H. pylori infection.     

降低大鼠心肌梗死模型死亡率的方法

目的 探讨降低大鼠心肌梗死模型死亡率的影响因素.方法 结扎180只Wistar雄性大鼠左冠状动脉前降支制备心肌梗死模型,分析麻醉药品、气管插管方法、结扎部位的高低及术后护理等因素对大鼠死亡率的影响,并取心脏组织行HE染色、氯化四唑（tetrazolium chloride,TTC）染色确认模型成功建立.结果 水合氯醛麻醉死亡率15%,戊巴比妥钠麻醉死亡率35%,差异有统计学意义（P＜0.05）;经口气管插管大鼠死亡率10%,经气管切开插管死亡率29.4%,差异有统计学意义（P＜0.05）;有术后护理大鼠死亡率5.9%,无术后护理大鼠死亡率22.2%,差异有统计学意义（P＜0.01）.而结扎部位的不同对大鼠手术死亡率及手术成功率亦有明显的影响.结论 本研究显示麻醉药品、气管插管方法、结扎部位的高低及术后护理等因素均对梗死模型的死亡率有影响,选择合适的麻醉药品和剂量以及气管插管方法,正确的结扎部位及适当的术后护理等可明显降低制作大鼠心肌梗死模型死亡率.