壳聚糖体内外抗幽门螺杆菌的实验研究

目的 研究壳聚糖体内外抗幽门螺杆菌（Hp）作用。方法 （1）采用打孔法检测了不同浓度、pH值、脱乙酰度壳聚糖及羧甲基壳聚糖在体外对蜘的抑菌作用。（2）建立BALB／c小鼠脚感染的动物模型后，随机分为8组：①对照组；②PPI组；③AM组；④AM＋PH组；⑤壳聚糖组；⑥壳聚糖＋PPI组；⑦壳聚糖＋AM组；⑧壳聚糖＋AM＋PPI组。分别给予上述药物每日2次灌胃，共2周。停药后4周，处死小鼠，无菌条件下取胃黏膜进行砷定量培养和石蜡包埋切片，进行Giemsa染色。结果 （1）壳聚糖和羧甲基壳聚糖在体外对3株邱标准菌株具有普遍的抑菌作用；在pH6-4范围内，随pH值降低，抗菌作用增强，差异有统计学意义（P〈0．01），最佳pH值为4；70％、88．5％脱乙酰度壳聚糖及羧甲基壳聚糖的抗伽作用差异有统计学意义（P〈0．05），抑菌强度依次为DD70壳聚糖、DD88．5，壳聚糖和羧甲基壳聚糖；在1％-5％浓度范围内羧甲基壳聚糖抗伽作用差异无统计学意义；在0．5％-2％浓度范围内70％、88．5％脱乙酰度壳聚糖抗伽作用差异也无统计学意义（P〉0．05）。（2）对饰感染的小鼠，以上8组的坤清除率分别为O％、O％、41．7％、58．3％、58．3％、66．7％、83．3％、91．7％，其中③-⑧组的坤清除率显著高于①和②组（P〈0．05），⑧组的坤清除率还显著高于③组（P〈0．05）。坤定植密度研究发现，各组之间坤定植密度差异有统计学意义（P〈0．001），饰定植密度在③一⑧组显著低于①和②组（P〈0．05）；⑥-⑧组显著低于③组（P〈0．05）；⑧组显著低于④组（P〈0．05）。结论 壳聚糖和其衍生物对坤有普遍的抑菌作用；壳聚糖及其衍生物的抗饰作用受多种因素影响，其中pH值对壳聚糖抗菌作用的影响最为明显，在pH值6—4范围内，壳聚糖的抗坤活性随着pH值的下降而显著增强；壳聚糖在体内有抗坤作用，并与AM有协同作用。     

以壳聚糖为佐剂的幽门螺杆菌蛋白抗原对幽门螺杆菌感染的免疫保护作用

目的探讨以壳聚糖为佐剂的幽门螺杆菌（脚）抗原对脚感染的免疫保护作用。方法BALB／c小鼠随机分为9组：（1）空白对照组：PBS溶液；（2）壳聚糖酸溶液组；（3）壳聚糖颗粒组；（4）印抗原组；（5）np抗原＋壳聚糖酸溶液组；（6）Hp抗原＋壳聚糖颗粒组；（7）Hp抗原＋CT组；（8）脚抗原＋壳聚糖酸溶液＋CT组；（9）Hp抗原＋壳聚糖颗粒＋CT组。各组于第0、7、14、21天灌胃各免疫1次，免疫后4周给予109CFUIml的SSlnp菌液0．5ml／只进行攻击，隔日1次，共2次。4周后，采用定量砌培养和病理改良Giemsa染色法检测胃黏膜内伽感染情况。结果（1）以壳聚糖为佐剂的印抗原的免疫保护率达60％，与以凹为佐剂的伽抗原的免疫保护率（58．33％）相似，显著高于单纯印抗原组及其他不含印抗原组（P〈0．05），同时以凹＋壳聚糖为佐剂的坳疫苗的保护率为84．62％、85．71％，高于CT或壳聚糖单独作佐剂组。（2）病理组织学检测含佐剂组的铷定植计分显著低于无佐剂组和无抗原组（P〈0．05），壳聚糖和CT联合应用组的跏定植计分最低，显著低于单以CT为佐剂组（P〈0．05）。（3）砌定量培养脚定植密度在佐剂中含壳聚糖组均显著低于对照组和单纯脚抗原组（P〈0．05），而单以CT为佐剂组脚定植密度与对照组和单纯坳抗原组差异无统计学意义（P〉0．05）。结论以壳聚糖为佐剂的坳抗原对Hp感染具有免疫保护作用，并且与CT有协同作用。     

以壳聚糖为佐剂的Hp疫苗诱导的体液免疫应答及其免疫保护效应

目的：探讨以壳聚糖为佐剂的Hp疫苗的免疫保护作用及其机制。方法:BALB/c小鼠随机分为9组：①空白对照组：PBS溶液；②壳聚糖酸溶液组；③壳聚糖颗粒组；④Hp抗原组；⑤Hp抗原+壳聚糖酸溶液组；⑥Hp抗原+壳聚糖颗粒组；⑦Hp抗原+CT组；⑧Hp抗原+壳聚糖酸溶液+CT组；⑨Hp抗原+壳聚糖颗粒+CT组，各组于第0、7、14、21 d 灌胃各免疫1次，免疫后4周给予1×1012CFU/L的SS1 Hp菌液每只 0.5 mL进行攻击，隔日1次，共2次。4周后，采用定量Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用ELISA法检测血清抗Hp IgG、IgG1、IgG2a及唾液和胃黏膜内抗Hp IgA，用SP免疫组织化学法检测胃黏膜内分泌型IgA（sIgA）。结果:①以壳聚糖为佐剂的Hp疫苗的免疫保护率达60%，与以CT为佐剂的Hp疫苗的免疫保护率（58.33%）相似，显著高于单纯Hp抗原组及其它不含Hp抗原组（P<0.01或P<0.05），同时以CT＋壳聚糖为佐剂的Hp疫苗的保护率为84.62%、85.71%，其Hp的定植评分显著低于无佐剂组及以CT为佐剂组（P<0.01，P<0.05）。②含佐剂的Hp疫苗所诱导产生的Hp IgG水平显著高于对照组及无佐剂组（P<0.01，P<0.05），而以CT＋壳聚糖为佐剂组所产生的抗Hp IgG水平显著高于仅以 CT或壳聚糖为佐剂组（P<0.05）。③胃黏膜内sIgA及特异性抗Hp IgA水平在壳聚糖为佐剂组与以CT为佐剂组无差别（P>0.05），显著高于无佐剂组，而壳聚糖与CT联合应用组显著高于单以CT为佐剂组（P<0.01，P<0.05）。结论:以壳聚糖为佐剂的Hp疫苗对Hp感染具有免疫保护作用，并可成功诱导黏膜局部的特异性体液免疫应答，从而发挥免疫防御作用。     

Th免疫反应在以壳聚糖为佐剂的幽门螺杆菌疫苗免疫保护中的作用

目的：探讨以壳聚糖为佐剂的脚疫苗的免疫保护作用及其机理。方法：BALB／c小鼠随机分为7组：空白对照组、壳聚糖酸溶液组、壳聚糖颗粒组、却抗原组、脚抗原＋壳聚糖酸溶液组、却抗原＋壳聚糖颗粒组和脚抗原＋CT组，各组于第0、7、14和21天灌胃各免疫1次，末次免疫后4周给予SS1Hp菌攻击，隔日1次，共2次。在攻击前后分批处死小鼠，取胃黏膜检测坳和Th1、Th2细胞因子含量，同时检测血清中抗Hp IgG2a和IgG1含量。结果：①以壳聚糖为佐剂的坳疫苗的免疫保护率达60％，与以CT为佐剂的印疫苗的免疫保护率（58．33％）相似，显著高于单纯脚抗原组及其他不含Hp抗原组（P〈0．001～0．05）。②却攻击后胃黏膜内IFN-γ、IL-2和IL-12含量在含佐剂组显著高于无抗原和无佐剂组（P〈0．001～0．05）；③坳攻击后胃黏膜内IL-4含量在以壳聚糖颗粒为佐剂组显著高于以CT为佐剂组（P〈0．05），以壳聚糖溶液为佐剂组显著高于对照组、无佐剂组及佐剂中含CT组（P〈0．001～0．05）。结论：以壳聚糖为佐剂的坳疫苗对脚感染具有免疫保护作用，同时可促进Th1和Th2的混合免疫反应。     

烫伤对大鼠免疫功能的影响及药膳饮食对其的调理作用观察

目的观察烫伤对大鼠免疫功能的影响及药膳饮食对其的调理作用。方法健康Wistar大鼠70只，体质量（200±20）g，雌雄各半，随机分成正常对照组（A组，n=10）、烧伤常规喂养组（B组，n=30）和烧伤药膳喂养组（C组，n=30）。A组37℃造成假伤后全部处死取材，B、C组大鼠造成30％TB-SAⅢ度烧伤伤后B组常规饲养，C组伤后2h开始以复温至37℃的药膳饮食灌胃，2ml/次，2次／d＋常规饲养。B、C组于伤后3、7、14d各取10只，无菌条件下取材送检，观察T淋巴细胞亚群和NK细胞活性、血浆IgA、IgG、IgM、C3、C4、含量、肠道sIgA含量的变化。结果①B、C组伤后CD3^＋、CD4^＋、CD4^＋／CD8^＋、NK细胞活性、IgA、IgG、IgM、C3、C4水平及肠道sIgA含量均低于A组，CD8^＋高于A组（P〈0．05或P〈0．01）；②C组与B组比较，各免疫指标均恢复快（P〈0．05或P〈0．01）。结论严重烫伤大鼠细胞免疫和体液免疫功能发生了改变；药膳饮食可以改善烫伤大鼠T淋巴细胞亚群分布，提高NK细胞活性，促进血浆IgA、IgM、IgG、C3、C4的恢复和肠黏膜细胞sIgA的分泌，从而改善机体的免疫功能。     

幽门螺旋杆菌感染对反流性食管炎发病和治疗的影响

目的探讨幽门螺旋杆菌（HP）感染对反流性食管炎（RE）发病和治疗的影响。方法收集西安市中心医院2011年6月至2013年6月的门诊病人642例，记录其性别、年龄、体重、反流性食管炎分级及HP感染严重程度。结果RE组HP（＋）感染率明显高于HP（-），差异有统计学意义（P〈0．05），而非RE组HP（＋）感染率与HP（-）间无明显差异（P〉0．05）。b组（C—D级）患者HP感染（＋＋／＋＋＋）明显高于a组（A—B级），差异有统计学意义（P〈0．05）。A组（HP根除＋PPI）与B组（PPI）GERD—Q评分、心理SF-36评分和生理SF-36评分相比无明显差异（P〉0．05）。结论HP感染与反流性食管炎的发病可能存在一定的关联，根除HP未能明显改善反流性食管炎的预后。     

慢性乙型肝炎患者外周血淋巴细胞CD8+CD38+的检测意义

目的：通过测定慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的表达及门冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、凝血酶原时间（PT），旨在为治疗慢性乙型肝炎提供有用的参考指标。方法：流式细胞术检测38例慢性乙型肝炎、14例肝硬化患者外周血淋巴细胞CD8^＋CD38^＋细胞的百分率；同时测定AST、TBIL和PT。结果：（1）慢性乙型肝炎组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．01，P〈0．01），肝硬化组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．05，P〈0．05）。肝硬化组CD8^＋CD38^＋、CD8^＋细胞均明显低于慢性乙型肝炎组（P〈0．05，P〈0．05）。（2）慢性乙型肝炎轻、中、重各组之间比较，中度组CD8^＋CD38^＋高于轻度组（P〈0．05），重度组CD8^＋CD38^＋、CD8^＋、CD38^＋均高于轻度组（P〈0．05，P〈0．05，P〈0．05）；重度组CD38^＋高于中度组（P〈0．05）；肝硬化组CD8^＋CD38‘均明显低于轻、中、重各组（P〈0．05，P〈0．01，P〈0．01），肝硬化组CD8^＋低于重度组（P〈0．01）。（3）慢性乙型肝炎患者AST、TBIL、PT不正常组CD8^＋CD38^＋均高于AST、TBIL、PT正常组。结论：慢性乙型肝炎患者CD8^＋CD38^＋细胞明显升高，表明慢性乙型肝炎患者细胞免疫处于异常激活状态，并与肝功能损伤有一定的相关性。慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的测定，对病情分析和诊断有一定的临床参考价值。     

川崎病患儿血清抗血管内皮细胞抗体检测的临床意义

目的探讨抗血管内皮细胞抗体（AECA）检测在川崎病（KD）早期诊断及体液免疫状况方面的临床意义。方法检测58例KD急性期患儿和43例对照组（其中普通发热对照23例，健康对照20例）儿童的血清AECA-IgG、免疫球蛋白IgG、IgA、IgM和补体C3、C4，计算各组AECA的阳性率，进而分别比较AECA阳性KD组、AECA阴性KD组与对照组的上述指标的浓度差异。结果①KD患儿AECA—IgG的阳性率为39．7％，明显高于健康对照组的5．0％（P〈0．01），也高于发热对照组的17．4％，但无统计学差异（P〉0．05）；②AECA阳性组与AECA阴性组IgG、IgM、IgA、C3浓度均分别明显高于发热对照组和正常对照组（P〈0．05），C4水平只有AECA阳性组明显高于发热对照组和健康对照组（P〈0．05），但AECA阳性组与阴性组间上述指标差异无统计学意义（P〉0．05）。结论血清抗内皮细胞抗体在川崎病患儿中有较高的阳性率，具有一定的早期辅助诊断价值。川崎病急性期存在着明显的体液免疫异常，IgA介导的体液免疫可能在其中扮演着重要角色。     

重组耻垢分枝杆菌制剂预防幽门螺杆菌感染的作用机理的初步探讨

目的探讨重组耻垢分枝杆菌实验制剂（recombinant Mycobacterium smegmatis，rMs）预防幽门螺杆菌（Helicobacter pylori，Hp）感染的作用机理。方法实验制剂免疫BLAB／c小鼠4周，用Hp攻击后4周，取血清、胃、脾组织，RT-PCR检测小鼠胃黏膜和脾组织的TH1型细胞因子（IFN-γ、IL-2、IL-12）与TH2型细胞因子（IL-4、IL-6、IL-10）；用ELISA检测针对唧的特异性血清IgG、IgG1、IgG2a和IgA水平；用免疫组化方法检测胃黏膜固有层IgA表达；用MTT检测脾淋巴细胞增殖。结果免疫后BALB／c小鼠特异性抗体显示rMs制剂诱导Hp特异性血清IgG、IgA水平都升高，以IgG2a占优势。用Hp抗原刺激后各免疫组小鼠脾淋巴细胞均有明显增殖。免疫组化显示各免疫组小鼠胃黏膜固有层IgA阳性标记腺体数明显高于对照组。RT-PCR证实，跏攻击之前，各免疫组胃和脾组织已出现了IFN-γ、IL-12、IL-2表达而无IL-4、IL-6、IL-10表达。PBS对照组和单纯耻垢分枝杆菌（Ms）组胃黏膜和脾组织各细胞因子均无表达；Hp攻击后，PBS和单纯Ms组胃组织出现以TH1细胞IFN-γ为主的增生，IFN-γ水平显著高于rMs组（P〈0．05）；而3个rMs制剂组脾脏则出现以TH2细胞（IL-4）为主的增生，IL-4水平显著高于对照组（P〈0．05）。提示该制剂的作用机制是诱导TH1，TH2平衡型免疫应答。结论成功地建立了BALB／c小鼠的Hp感染模型，对rMs制荆的免疫保护机理研究结果显示，rMs能产生以TH1细胞和TH2细胞协同作用的保护性免疫应答。     

恶性肿瘤患者神经-内分泌-免疫网络有关指标的紊乱及其临床意义

目的：初步分析恶性肿瘤患者神经-内分泌-免疫网络紊乱情况及其临床意义。方法：102例经病理明确诊断的恶性肿瘤患者，用放射免疫法和流式细胞技术检测外周血中CD3^＋、CD4^＋和CD8^＋细胞数量、NK比值，以及IL-1β、IL-6、TNF-α、ACTH、CORT和DA及NE水平。结果：（1）在几乎所有类型肿瘤患者中都表现为CD3^＋、CD4^＋细胞数量下降（P〈0．05或P〈0．01）、CD8^＋的升高以及CD4^＋／CD8^＋的降低（P〈0．05或P〈0．01）。NK细胞在各恶性肿瘤组较正常均有下降（P〈0．05）。肿瘤患者的IL-1β和TNF-α较正常均有不同程度的下降。与正常对照相比，IL-6在肿瘤患者中明显升高（P〈0．05）；各类型肿瘤间无明显差异（P〉0．05）。ACTH在各组间无差异。皮质醇在消化道肿瘤患者中最高；明显高于脑瘤和肺癌患者（P〈0．05）。DA和NE在大多数类型的恶性肿瘤患者组均升高（P〈0．05或P〈0．01）。（2）化疗和放疗患者组CD3,细胞数量较无化放疗者明显降低（P〈0．05）。放疗患者组的CD8^＋升高及CD4^＋／CD8^＋比值下降较无放化疗组和化疗组明显，与无放化疗相比有统计学意义（P〈0．05）。放疗患者IL-1β和IL-6值均升高，高于化疗者和无放化疗者（P〈0．05）；化疗患者TNF—α值较正常升高（P〈0．05），与放疗者无明显差异。放疗患者组的DA水平低于无放化疗患者组（P〈0．05）。（3）在女性肿瘤患者中，CD3^＋、IL-1β和TNF-α水平均高于男性（P〈0．05）。（4）在恶性肿瘤包括有远处转移患者中，IL-6明显升高（P〈0．05）。（5）一般状况差（PS：3～4级）患者的CORT和TNF—d升高明显（P〈0．05）。而临床较晚期（Ⅲ～Ⅳ期）患者的CD4^＋／CD8^＋比值下降明显（P〈0．05）。结论：恶性肿瘤患者神经．内分泌-免疫网络的紊乱，以皮质醇、TNF—α和IL-6及DA、NE水平不同程度的升高、以及CD3^＋、CD4^＋和CD4^＋／CD8^＋细胞数量不同程度的减少为主要表现，其与肿瘤患者的性别、病理类型、转移与否及其治疗方式等有关；与肿瘤患者的一般状况无关。     

以壳聚糖为佐剂的幽门螺杆菌疫苗诱导的细胞免疫反应

目的:探讨以壳聚糖为佐剂的幽门螺杆菌(Hp)疫苗诱导的细胞免疫反应及其在免疫保护中的作用。方法:BALB/c小鼠随机分为空白对照组(PBS溶液)、壳聚糖酸溶液组、壳聚糖颗粒组、Hp抗原组、Hp抗原 壳聚糖酸溶液组、Hp抗原 壳聚糖颗粒组、Hp抗原 霍乱霉素(CT)组、Hp抗原 壳聚糖酸溶液 CT组及Hp抗原 壳聚糖颗粒 CT组,各组于第0、7、14、21天灌胃各免疫1次,末次免疫后4周给予1×1012CFU/L的SS1Hp菌液0·5mL/只进行攻击,隔日1次,共2次。在攻击前后分批处死小鼠,采用Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用定量ELISA法检测胃黏膜内IL-2、IL-4和IL-10含量;HE染色进行胃黏膜病理学检测。结果:①以壳聚糖为佐剂的Hp疫苗的免疫保护率达60%,与以CT为佐剂的Hp疫苗的免疫保护率(58·33%)相似,显著高于单纯Hp抗原组及其他不含Hp抗原组(P<0·05)。②胃黏膜内IL-2的水平攻击后含佐剂组显著高于对照组(P<0·05)。IL-10水平攻击前以壳聚糖为佐剂组高于无佐剂组,攻击后以CT 壳聚糖颗粒为佐剂组高于其他组(P<0·05)。IL-4水平攻击前以壳聚糖为佐剂组高于无佐剂组(P<0·05),攻击后以壳聚糖颗粒为佐剂组高于以CT为佐剂组(P<0·05),以壳聚糖溶液为佐剂组高于对照组、无佐剂组及佐剂中含CT组(P<0·05)。③胃黏膜炎症程度在单纯壳聚糖组和以壳聚糖颗粒为佐剂组显著低于以CT为佐剂组(P<0·05)。结论:①以壳聚糖为佐剂的Hp疫苗对Hp感染具有免疫保护作用。②以壳聚糖为佐剂的Hp疫苗可促进Th1和Th2的混合免疫反应,并逆转Hp感染所致Th2反应的抑制,使Th1和Th2反应达到平衡,从而发挥其免疫保护作用。③以壳聚糖为佐剂的Hp疫苗所致的免疫后胃炎显著低于以CT为佐剂的Hp疫苗。     

Intramucosal gastric adenocarcinoma of poorly differentiated type in the young is characterized by Helicobacter pylori infection and antral lymphoid hyperplasia.

The aim of this investigation was to clarify the histological characteristics of gastric cancer in the young. Twenty-three surgically resected specimens of young patients (under 30 years of age; young group) with intramucosal cancer of poorly differentiated type and 42 surgically resected specimens of elderly patients (more than 40 years of age; elderly group) with tumors of the identical depth and histological type were examined. The degree of gastritis and Helicobacter pylori (H. pylori) infection was evaluated according to the updated Sydney system. The incidence of H. pylori infection was significantly higher in the young group than in the elderly group (96 vs 36%, P<0.05). Within the background mucosa, antral chronic inflammatory infiltrates with lymphoid-follicle hyperplasia were more severe, and intestinal metaplasia was less frequent in the young group than in the elderly group. Glandular atrophy was not different between the two groups. Intramucosal gastric adenocarcinomas of poorly differentiated type in the young may be associated with H. pylori infection with antral chronic inflammation with lymphoid-follicle hyperplasia, regardless of the existence of intestinal metaplasia within the background gastric mucosa.     

幽门螺杆菌对体外感染胃黏膜上皮细胞TGF-β1 B7-H1表达的影响

目的 探讨适当菌量幽门螺杆菌(H.pylori)对体外感染胃黏膜上皮细胞转化生长因子β1(TGF-β1)、B7-H1 mRNA表达的影响及其诱导TGF-β1表达的菌体因素,为支持H.pylori通过诱导胃黏膜上皮细胞表达TGF-β1及B7-H1,抑制宿主免疫功能,参与H.pylori免疫逃逸提供依据.方法 (1)选用1.0×109CFU/ml(低浓度)、4.0×109 CFU/ml(中浓度)、8.0×109 CFU/ml(高浓度)H.pylori国际标准毒力菌株NCTC 11637活菌悬液分别感染体外培养胃黏膜上皮细胞,建立不同共孵育时间(0 h、0.5 h、1 h、1.5 h、2 h、4 h、8 h、12 h)H.pylori体外感染细胞模型,设立未加H.pylori的胃黏膜上皮细胞对照组,以酶联免疫吸附法(ELISA)检测感染及对照细胞各时间点培养上清TGF-β1含量,原位杂交法检测不同浓度感染及对照细胞共培养12 h B7-H1 mRNA的表达.(2)同时用H.pylori中浓度灭活菌悬液与体外培养胃黏膜细胞共孵育,测定共孵育细胞2 h、12 h培养细胞上清的TGF-β1含量.(3)用超声粉碎并经离心获取的H.pylori菌体成分上清、沉淀以及煮沸后上清、沉淀分别作用体外培养胃黏膜细胞,测定2 h、12 h培养细胞上清TGF-β1含量.结果 (1)H.pylori活菌悬液3种浓度各时间组胃黏膜上皮细胞培养上清TGF-β1含量均较对照组明显增高(P<0.05),各浓度时间组TGF-β1表达量有栩似的动态趋势,但尤以中浓度组TGF-β1表达量最高(P<0.05).(2)中浓度H.pylori灭活菌株组与同浓度活菌组细胞培养上清TGF-β1含量差异无统计学意义(P>0.05).(3)H.pylori菌体成分上清组较对照组和沉淀组细胞培养上清TGF-β1表达增高(P<0.01);上清煮沸组较未煮沸组明显降低(P<0.01);沉淀煮沸组与未煮沸组差异无统计学意义(P>0.05).(4)高、中、低浓度组感染细胞12 h B7-H1 mRNA的表达均较对照组增高(P<0.05),以中浓度组表达最高,并与高、中、低浓度组12 h感染细胞上清TGF-β1含量呈正相关(rs=0.628,P<0.01).结论 H.pylori可直接诱导胃黏膜上皮细胞分泌TGF-β1,其诱导因素为可溶性不耐热菌体成分;H.pylori同样可诱导胃黏膜上皮细胞B7-H1 mRNA表达增高,且B7-H1 mRNA表达与TGF-β1呈正相关.推测H.pylori通过诱导TGF-β1、B7-H1高表达,抑制宿主免疫应答,参与H.pylori免疫逃逸.     

HP1188-IgY对感染幽门螺杆菌BALB/c小鼠的治疗作用

目的:评价抗幽门螺杆菌(Helicobacter pylori,H.pylori)HP1188蛋黄抗体(HP1188-immunoglobulin yolk,HP1188-Ig Y)对BALB/c小鼠胃内H.pylori感染的治疗效果。方法:建立H.pylori感染BALB/c小鼠的动物模型,将建模成功的80只小鼠随机分为8组,每组10只。第1组(抗生素治疗组)、第2组(1 mg HP1188-Ig Y)、第3组(1 mg HP1188-Ig Y+30%硫糖铝)、第4组(5 mg HP1188-Ig Y)、第5组(5 mg HP1188-Ig Y+30%硫糖铝)、第6组(PBS对照组)和第7组(30%硫糖铝对照组)分别连续治疗10 d,每天1次;第8组间隔48 h皮下注射2.5 mg HP1188-Ig Y,共2次;另10只正常小鼠为第9组(健康对照组)。处理完成2周后取各组小鼠胃组织作H.pylori培养、快速尿素酶试验、PCR检测H.pylori特异性vac A和cag A及病理组织学检查,观察H.pylori清除情况并进行评分。结果:在小鼠体内,灌胃(1 mg HP1188-Ig Y+30%硫糖铝)即可有效治疗H.pylori感染引起的胃黏膜损害,其治疗效果与抗生素相似。结论:成功构建了H.pylori感染的BALB/c小鼠模型,选择30%硫糖铝为抗体保护剂,经口服HP1188-Ig Y可抑制小鼠胃内H.pylori感染。     

K-ras mutations and cell kinetics in Helicobacter pylori associated gastric intestinal metaplasia: a comparison before and after eradication in patients with chronic gastritis and gastric cancer

BACKGROUND: Helicobacter pylori related gastric intestinal metaplasia (IM) is considered to be a precancerous lesion. AIMS: To identify the effects of H pylori eradication on K-ras mutations, cell kinetics in IM and histological changes in patients with and without gastric cancers in a one-year prospective study. METHODS: Patients included group A (n = 39), chronic gastritis, and group B (n = 53), intestinal-type early gastric cancer patients who had all undergone endoscopic mucosal resection (n = 25) or surgical resection (n = 28). K-ras codon 12 mutations in IM were examined, followed by DNA sequencing analysis. Proliferating and apoptotic cells were detected with anti-Ki-67 antibody and using the TUNEL method, respectively. RESULTS: The incidence of K-ras mutations in the cancer was only 3.8%. The mutant K-ras in IM was observed more frequently in group A (46.2%) than in group B patients (1.9%) (p<0.005). After eradication, the K-ras mutations significantly declined to 12.8% in group A (p<0.005). The mutation pattern of K-ras codon 12 before eradication was that GGT was mainly changed to AGT (50%) in group A. AGT transformation was not affected by treatment. Apoptosis in IM showed an increase after H pylori eradication in both groups (p<0.05 in group A) although no histological improvement in IM was observed. The monocyte score was significantly higher in group A than in group B (p<0.05); the score improved significantly after eradication. CONCLUSIONS: K-ras mutations in IM do not always play a role in gastric carcinogenesis but cell kinetics, especially apoptosis, in IM may contribute to it. There are early events in K-ras mutations which are influenced by H pylori infection; some mutations may also be selected by eradication. These unstable K-ras mutations in IM may be related to lymphocyte infiltration caused by H pylori infection.     

Expression of Lewis antigens and their precursors in gastric mucosa: relationship with Helicobacter pylori infection and gastric carcinogenesis

Lewis (Le)-associated antigens are carbohydrates that are related biochemically to the ABO blood groups, and may have a role in Helicobacter pylori adherence. To evaluate their relationship to clinicopathological outcomes, gastric Le expression, including type 1 precursor, type 1 H, Le(a), Le(b), Le(x), Le(y) and sialylated Le(a) (CA19-9), was evaluated immunohistochemically in 233 gastric biopsy specimens obtained at routine gastroscopy. Expression was also investigated in gastric tissues showing chronic gastritis, intestinal metaplasia, and carcinoma from 42 patients with gastric cancer. A polymerase chain reaction was performed for H. pylori and the bacterial babA2 gene. We identified type 1 precursor expression in 34.3%, type 1 H in 55.8%, Le(a) in 44.2%, Le(b) in 82.0%, Le(x) in 44.2%, Le(y) 56.7%, and CA19-9 in 16.3% of the 233 gastric biopsy specimens. Expression of type 1 H, Le(b), and CA19-9 was significantly associated with H. pylori infection and histological features (p < 0.05), and expression of type 1 H was an independent predictive factor for H. pylori infection by multivariate logistic regression (p = 0.020). Positivity for the babA2 genotype correlated significantly with H. pylori infection and type 1 H expression in gastric biopsy specimens (p < 0.05). The babA2 genotype was more frequent in gastric mucosa from the gastric cancer patients than in gastric biopsy specimens from routine gastroscopy (p = 0.009). In the 42 gastric cancer patients, the frequency of type 1 precursor, Le(a), and Le(x) expression was significantly higher in intestinal metaplasia and carcinoma than in chronic gastritis (p < 0.05), but the frequency of type 1 H and Le(b) expression was significantly lower in intestinal metaplasia and carcinoma (p < 0.05). In conclusion, Le expression, especially that of type 1 H, was significantly associated with clinicopathological features. In gastric cancer patients, Le expression was altered in intestinal metaplasia and carcinoma in comparison with chronic gastritis.     

Helicobacter pylori infection and chronic gastritis in gastric cancer.

AIMS: To investigate the prevalence of Helicobacter pylori associated chronic gastritis in patients with gastric cancer. METHODS: Serum IgG antibodies for H pylori were determined in 54 consecutive patients with gastric carcinoma. The prevalence of H pylori in gastric mucosa was also examined histologically (modified Giemsa) in 32 patients from whom adequate biopsy specimens of the antrum and corpus were available. Thirty five patients with gastrointestinal tumours outside the stomach and 48 with non-gastrointestinal malignancies served as controls. RESULTS: Of the 54 patients, 38 (70%) had H pylori antibodies (IgG) in their serum (three additional patients had H pylori antibodies IgA, class specific but not IgG specific). This prevalence was significantly higher (p less than 0.05) than that (49%) in the 35 controls. No differences in prevalence of H pylori antibodies were found between gastric cancer cases of intestinal (IGCA) or diffuse (DGCA) type, both these types showing H pylori antibodies (IgG) in 71% of the patients. In the subgroup of 32 subjects, five patients had normal gastric mucosa and four showed corpus limited atrophy ("pernicious anaemia type" atrophy of type A). All of these nine patients had no evidence of current or previous H pylori infection in serum (no IgG antibodies) or in tissue sections (negative Giemsa staining). The remaining 23 patients had antral or pangastritis, and all had evidence of current or previous H pylori infection. CONCLUSIONS: H pylori associated chronic gastritis was the associated disease in 75% of the patients with gastric cancer occurring equally often in both IGCA and DGCA groups. About 25% of cases seem to have a normal stomach or severe corpus limited atrophy, neither of which showed evidence of concomitant H pylori infection.     

IgE-containing cells in gastric mucosa with and without Helicobacter pylori infection

Gastric mucosa responds with inflammation to Helicobacter pylori (H. pylori) infection. While numerous reports have shown that the immune system produces specific IgG, IgA, and IgM isotype anti H. pylori antibodies, IgE-mediated pathways of H. pylori-associated gastritis are mostly unknown. Our aim was to evaluate whether an increased presence of IgE in the antral gastric mucosa is responsible for the severity of the H. pylori-associated gastritis. The number of IgE-containing cells was estimated in formalin-fixed, paraffin-embedded antral gastric biopsy specimens using immunohistochemistry in three groups of patients: (i) 20 H. pylori-positive cases with moderate inflammation, (ii) 19 H. pylori-negative cases with moderate inflammation, and (iii) 19 H. pylori-negative cases with normal mucosa. In chronic gastritis, the number of IgE-positive cells increased significantly as compared to normal mucosa. In gastritic patients, H. pylori positivity was accompanied by a significant accumulation of IgE-positive cells, mainly plasma cells. These data suggest that IgE-mediated immune response probably plays an important role in the development of H. pylori-associated gastritis.