MNNG诱导人胃粘膜上皮细胞的损伤作用

目的探讨甲基硝基亚硝基胍（ＭＮＮＧ）对胃粘膜上皮细胞的损伤作用．方法对ＭＮＮＧ在诱发体外培养的人胎胃粘膜上皮细胞的非程序ＤＮＡ合成、脂质过氧化和ｒａｓＰ２１蛋白进行观察．结果ＭＮＮＧ处理组各时相胃粘膜上皮细胞的增殖速率，非程序ＤＮＡ合成水平（１８３８ｃｐｍ±２０５ｃｐｍｖｓ７０５ｃｐｍ±１２５ｃｐｍ，Ｐ＜００１），质脂过氧化物（第２０天，７．４２ｎｍｏｌ／ｍｇ±０．６８ｎｍｄ／ｍｇｖｓ３．６２ｎｍｏｌ／ｍｇ±０．５４ｎｍｏｌ／ｍｇ蛋白Ｐ＜０．０１）和ｒａｓＰ２１蛋白含量（第４０天，１２１Ａ±０１２Ａｖｓ０５１Ａ±００７Ａ，Ｐ＜００１）．结论ＭＮＮＧ可诱发胃粘膜上皮细胞恶性转化，其作用机制可能是通过损伤细胞ＤＮＡ，诱发质脂过氧化或导致ｒａｓＰ２１异常表达所致．     

冬虫夏草多糖治疗慢性丙型肝炎患者的临床研究

目的研究冬虫夏草多糖（ＣＰ）治疗慢性丙型肝炎的疗效．方法慢性丙型肝炎患者２１例，口服ＣＰ１５ｍＬ，３次／ｄ，连服３ｍｏ，治疗前后检测肝功能、血清肝纤维化标志物外周血Ｔ细胞亚群及ＮＫ活性的变化．结果慢性丙型肝炎患者经ＣＰ治疗后，血清ＡＬＴ（Ｕ／Ｌ，６１±３５ｖｓ３５±１５）及ｒＧＴ（Ｕ／Ｌ，１６９±８５ｖｓ１１８±５２）较治疗前显著降低（Ｐ＜００５）．血清ＨＡ（μｇ／Ｌ，２９３±１０９ｖｓ２１４±９６）、ＰⅢＰ（μｇ／Ｌ，１４３±４８ｖｓ１１４±４２）及ＣⅣ（μｇ／Ｌ，２４５±９８ｖｓ１８８±８７）均较治疗前显著下降（Ｐ＜００１，＜００５及＜００５）；ＣＤ４（３６４％±６６％ｖｓ４１０％±５６％）、ＣＤ４／ＣＤ８（１１４±０４０ｖｓ１４３±０２２）、ＮＫ（１６７％±４６％ｖｓ１９７％±４２％）均较治疗前显著增加（Ｐ＜００１），而ＣＤ８（３２６％±４７％ｖｓ２８９％±３７％）则明显降低（Ｐ＜００５）；血清胆红素略减、清蛋白略增但差异均无显著性．结论冬虫夏草多糖可以增强慢性丙型肝炎患者细胞免疫功能，改善肝功能，并具有一定的抗纤维化作用     

根除Hp前后胃粘膜多胺含量变化的意义

目的探讨成功根除幽门螺杆菌（Ｈｐ）感染后，胃粘膜组织学及多胺（腐胺、精脒和精胺）含量变化的意义．方法Ｈｐ阳性患者５６例，其中男３９例，女１７例；胃溃疡７例，十二指肠溃疡１８例，慢性胃炎３１例．根除Ｈｐ前和根除治疗４ｗｋ～６ｗｋ后胃粘膜多胺含量及其组织学变化，同时设Ｈｐ阴性患者２０例作对照．结果Ｈｐ阳性患者胃窦粘膜蛋白多胺含量（μｍｏｌ／ｇ）明显高于阴性患者（腐胺０９６±０４９ｖｓ０４１±０３１，精脒２０１±０４８ｖｓ０８３±０３７，精胺２５８±０５７ｖｓ１３３±０４２，Ｐ＜００１）．Ｈｐ根除后就显著下降（腐胺０４８±０４０ｖｓ０９４±０４２，精脒１１１±０３４ｖｓ１９８±０３９，精胺１５３±０５５ｖｓ２６６±０４４，Ｐ＜００５）．胃窦部粘膜腐胺和精脒含量高于胃体部（腐胺０９９±０３１ｖｓ０５９±０２４，精脒２０７±０３８ｖｓ１３０±０３４，Ｐ＜００５）．组织学检查显示根除Ｈｐ感染后炎细胞浸润程度明显降低．结论根除Ｈｐ感染能够降低胃粘膜的增殖活性，从而降低胃癌的危险性．     

大肠癌患者促胃液素检测的临床意义

目的研究大肠癌患者血清及癌细胞内促胃液素（Ｇａｓ）水平及临床意义．方法采用ＲＩＡ法检测３５例大肠癌患者血清和癌细胞及癌旁粘膜细胞内Ｇａｓ水平．结果大肠癌患者术前、术后血清Ｇａｓ水平与对照组无显著差异（Ｐ＞００５），根治术后明显低于术前（２９ｎｇ／Ｌ±５ｎｇ／Ｌｖｓ３５ｎｇ／Ｌ±１２ｎｇ／Ｌ，ｔ＝２７７２，Ｐ＜００１），在高分化（３６ｎｇ／Ｌ±１６ｎｇ／Ｌｖｓ２８ｎｇ／Ｌ±５ｎｇ／Ｌ）和中分化腺癌组中（３８ｎｇ／Ｌ±７ｎｇ／Ｌｖｓ２７ｎｇ／Ｌ±３ｎｇ／Ｌ）差异显著（ｔ＝２１５２和２３５６，Ｐ＜００５）．大肠癌细胞内Ｇａｓ水平明显高于癌旁３ｃｍ和６ｃｍ粘膜（２１３ａｇ／细胞±７２ａｇ／细胞ｖｓ１４７ａｇ／细胞±３６ａｇ／细胞，１３９ａｇ／细胞±３２ａｇ／细胞；ｔ＝４８９１和５６１３，Ｐ＜００１）和正常粘膜（１３６ａｇ／细胞±４６ａｇ／细胞；ｔ＝２５３４，Ｐ＜００５），高分化腺癌明显高于低分化和粘液腺癌（２４１ａｇ／细胞±７８ａｇ／细胞ｖｓ１６１ａｇ／细胞±４６ａｇ／细胞，ｔ＝２５０５，Ｐ＜００５）．结论大肠癌细胞可通过自分泌方式分泌Ｇａｓ，Ｇａｓ升高是大肠癌分化良好的标志     

胃癌及消化性溃疡患者胃窦粘膜胃肠激素的变化

目的探讨胃癌及消化性溃疡（ＰＵ）患者胃窦粘膜胃肠激素变化的意义．方法内镜及活检确诊的浅表性胃炎（ＣＳＧ）１０例，胃溃疡（ＧＵ）１５例，十二指肠溃疡（ＤＵ）１２例，胃癌（ＧＣ）６例．胃镜下取胃窦粘膜，用ＲＩＡ法测定胃泌素（Ｇａｓ）、生长抑素（ＳＳ）、Ｐ物质（ＳＰ）的含量，各组间进行比较．结果胃窦粘膜ＳＳ含量在ＧＵ，ＤＵ，ＣＳＧ，ＧＣ组分别为２５１ｐｇ／ｍｇ±１９４ｐｇ／ｍｇ（以下同），４７０±１７９，５３２±２１１及１２９３±５２３。其中ＧＵ组低于其余各组（Ｐ＜００５），而ＧＣ时则显著升高（Ｐ＜００１）．ＳＰ含量在ＤＵ组显著降低，与ＧＵ，ＣＳＧ，ＧＣ比较分别为４７９±１５７ｖｓ７６５±４１５，７８９±３９０及８０１±３４６，Ｐ＜００５；ＧＣ患者Ｇａｓ水平显著高于ＣＳＧ组，为４６４５±２９４４，ｖｓ２７６８±１５７２，Ｐ＜００１．结论胃粘膜中Ｇａｓ，ＳＳ，ＳＰ含量的变化可能在ＰＵ及胃癌的发病机理中起重要作用．     

维甲酸对MNNG所致胃粘膜上皮细胞非程序DNA合成、脂质过氧化和rasp21表达的影响

目的和方法：对维甲酸预防甲基硝基亚硝基胍（ＭＮＮＧ）诱导体外培养胎胃粘膜上皮细胞恶性转化的作用进行研究。结果：结果发现，胃粘膜细胞先用１０－５Ｍ或１０－６Ｍ维甲酸预处理４小时，再给ＭＮＮＧ组非程序ＤＮＡ合成（ＵＤＳ）水平，质脂过氧化物和ｒａｓＰ２１含量，均显著低于ＭＮＮＧ组（Ｐ＜００５～００１）。结论：结果提示，维甲酸对ＭＮＮＧ诱导的胃粘膜上皮细胞损伤有防护作用     

老年胃酸分泌与生长抑素的研究

目的探讨老年胃泌酸功能与生长抑素的调节机制．方法我室应用ＲＩＡ法对２０例老年人和１５例青年人胃液及９只Ｗｉｓｔａｒ老年鼠，２０只青年鼠胃粘膜组织、血浆生长抑素含量进行测定，以及对老年人和青年人进行胃酸分泌试验．结果老年人组比青年组胃液生长抑素呈高水平分泌，空腹胃液ＳＳ为（１３６５±４４３）ｎｇ／Ｌｖｓ（８５９±３４５）ｎｇ／Ｌ，Ｐ＜００１；刺激后ＳＳ为（１９６７±６６４）ｎｇ／Ｌｖｓ（１１４１±１０７３）ｎｇ／Ｌ，Ｐ＜００１．老年人比青年人胃酸分泌量明显减少，ＢＡＯ为（３５±２１）ｍｍｏｌ／ｈｖｓ（６９±４４）ｍｍｏｌ／ｈ，Ｐ＜００１；ＰＡＯ为（１３６±６４）ｍｍｏｌ／ｈｖｓ（２０８±１１２）ｍｍｏｌ／ｈ，Ｐ＜００５．不同年龄Ｗｉｓｔａｒ大鼠胃粘膜组织生长抑素具有不同的生理浓度，在老龄鼠比青年鼠ＳＳ也呈高水平分布，为（４５２５±１８４２）ｎｇ／ｇ蛋白ｖｓ（１５０９±９３８）ｎｇ／ｇ蛋白，Ｐ＜００１．结论老年胃泌酸功能改变与生长抑素调节作用存在重要关系．     

胃肠激素在肝硬变胃粘膜病变发病机制中的作用

目的探讨胃肠激素（ＧＩＨ）在肝硬变胃粘膜病变发病机制中的意义．方法用ＲＩＡ法测定临床诊断为肝硬变伴胃粘膜病变（ＣＧＭＬ）１７例，肝硬变（ＬＣ）３２例，胃溃疡（ＧＵ）１０例，轻度浅表性胃炎（ＣＳＧ）２３例及正常对照者３６例的空腹血浆生长抑素（ＳＳ），血管活性肠肽（ＶＩＰ），胃动素（ＭＴＬ）及促胃液素（Ｇａｓ）的变化，并结合各疾病组的内镜特点，分析ＣＧＭＬ与某些ＧＩＨ的关系．结果血浆ＳＳ水平（ｎｇ／Ｌ）在ＣＧＭＬ及ＧＵ组分别为４１２±２１２及３７６±１４３，与正常对照组（６１２±１７０）、ＬＣ组（６６２±３０９），ＣＳＧ组（６４８±３９１）比较，Ｐ均＜００１；ＶＩＰ浓度（ｎｇ／Ｌ）则在ＣＧＭＬ组（６３４±３５８）、ＧＵ组（１１０９±４２４）、ＬＣ组（７７８±４２６）及ＣＳＧ组（６３４±３４７）均高于正常对照组（３２７±１４４），Ｐ均＜００１．ＣＧＭＬ，ＧＵ及ＣＳＧ具不同的内镜特点；ＣＧＭＬ的发病与食管静脉曲张（ＥＶ）关系密切，随ＥＶ程度的加重，发病率有增高的趋势．结论ＣＧＭＬ发病与门脉高压关系密切，激素代谢紊乱可能通过影响内脏血流及粘膜抗损害能力而参与其形成．     

发热对豚鼠胆汁成分的影响

目的研究发热对豚鼠胆汁成分及胆石形成的影响．方法将豚鼠６０只分为致石饲料组和普通饲料组，再将每组分为发热组和对照组．皮下注射煮沸脱脂牛奶１ｍｇ／ｋｇ使发热组动物发热，每周１次，共４次．４５ｄ后将６０只豚鼠全部处死，其中发热组在处死前３６ｈ皮下注射松节油１ｍｌ／ｋｇ，使其在处死前处于发热状态．检查胆囊并取胆汁进行分析．结果致石饲料发热组结石发生率最高，为４００％，而致石饲料不发热组为１３３％（Ｐ＜００１）．发热组胆汁总蛋白（２５ｇ／Ｌ±１９ｇ／Ｌｖｓ１０ｇ／Ｌ±０９ｇ／Ｌ，Ａ２ｖｓＡ１）及胆红素浓度（７４３μｍｏｌ／Ｌ±６２６μｍｏｌ／Ｌｖｓ１２３μｍｏｌ／Ｌ±５５μｍｏｌ／Ｌ，Ｐ＜００５）较对照组明显升高．结论发热对豚鼠胆汁成分的影响在胆石形成中具有重要作用．     

二甲基硅油对胃液中反流胆红素的影响

目的测定胃液中的胆红素浓度以评价二甲基硅油对胆汁反流的疗效．方法慢性胃炎患者６５例，其中伴胆汁反流４５例，进一步分成２组：治疗组（３０例）在测定前服二甲基硅油＋硫糖铝；对照组（１５例）单服硫糖铝．用胆红素氧化酶法测定胆红素浓度．结果无胆汁反流的２０名患者中，胃液中胆红素浓度分别检测１５例为０，５例为０４μｍｏｌ／Ｌ～０８μｍｏｌ／Ｌ；治疗组在服二甲基硅油前后均值分别为（１６０±１２０）μｍｏｌ／Ｌ和（６６±４９）μｍｏｌ／Ｌ（Ｐ＜００１），而对照组则分别为（１４６±１２３）μｍｏｌ／Ｌ和（１５９±１９８）μｍｏｌ／Ｌ（Ｐ＞００５）；胆红素浓度减少３０％以上者治疗组中占８００％，对照组中占２６７％，两组间有高度显著差异（χ２＝１２１００８．Ｐ＜００１）．结论测定胃液中胆红素有助于判断胆汁反流，并肯定二甲基硅油可减少胆汁反流，从而减轻胆汁反流性胃炎．     

胃十二指肠疾病患者骨矿物质含量的变化意义

目的研究慢性胃炎、胃十二指肠溃疡和胃癌患者骨矿物质含量变化的意义．方法用单光子吸收法，以桡骨长度中下１／３处为测定点，测定慢性胃炎（ｎ＝３１）、消化性溃疡（ｎ＝２６）、胃癌（ｎ＝９）及正常对照组（ｎ＝４３）的骨矿物质含量，并以骨矿含量（ＢＭＣ，ｇ／ｃｍ）及骨面密度（ＢＭＣ／ＢＷ，ｇ／ｃｍ２）表示，并进行比较．结果慢性萎缩性胃炎（ｎ＝１４），ＢＭＣ＝１００８±０２２８ｇ／ｃｍ）和十二指肠球部溃疡（ｎ＝１６），ＢＭＣ＝０９０４ｇ／ｃｍ±０２０５ｇ／ｃｍ，ＢＭＣ／ＢＷ＝０６５２ｇ／ｃｍ２±００８６ｇ／ｃｍ２）骨矿含量显著低于正常对照组（ｎ＝４３，ＢＭＣ＝１１７６ｇ／ｃｍ±０３４１ｇ／ｃｍ，ＢＭＣ／ＢＷ＝０７８２ｇ／ｃｍ２±０１３４ｇ／ｃｍ２，Ｐ＜００５）．结论萎缩性胃炎及十二指肠溃疡患者ＢＭＣ明显下降，需纠正．     

p53 C-myc和P-gp蛋白在胃癌细胞中表达

目的研究胃癌组织中ｐ５３和Ｃｍｙｃ的表达与多药耐药性（ＭＤＲ）的关系．方法应用ＬＳＡＢ免疫组织化学方法研究６７例（男４１例，女２６例，平均年龄４６±１５８岁）胃癌标本中ｐ５３，Ｃｍｙｃ和Ｐｇｐ的表达．结果本组胃癌中ｐ５３阳性３２例（４７８％），Ｃｍｙｃ阳性３７例（５５２％），Ｐｇｐ阳性３９例（５８２％）．淋巴结转移阳性胃癌ｐ５３阳性率（５６９％）和Ｃｍｙｃ阳性率（６４７％）显著高于淋巴结转移阴性的胃癌（Ｐ＜００５）．ｐ５３的异常表达与ｍｄｒ１基因表达呈显著正相关（ｒ＝０６３，Ｐ＜００５），而Ｃｍｙｃ和ｍｄｒ１的表达无明显相关．结论ｐ５３异常表达可增加ｍｄｒ１基因的表达，从而使胃癌细胞获得ＭＤＲ表型     

幽门螺杆菌感染患者胃氨的变化

目的探讨由Ｈｐ产生的氨在Ｈｐ致病机理中的作用．方法对１６例伴有Ｈｐ感染的消化性溃疡及慢性活动性胃炎患者，应用抗菌素对症治疗４周．分别在治疗前后用尿素酶试剂药盒检查Ｈｐ感染情况，并用Ｎｅｓｌｅｒ′ｓ显色法测定治疗前后的胃液氨浓度．结果治疗后有８例患者Ｈｐ阴转，胃氨浓度由治疗前的３０５７±１９６５μｍｏｌ／Ｌ降至１７６９±９７８μｍｏｌ／Ｌ（Ｐ＜００１），同时胃粘膜病变减轻或治愈．结论氨是Ｈｐ致病的重要因素之一．动态测定胃氨浓度可以间接反映Ｈｐ的感染情况，并可以作为判定治疗效果的一项指标．     

Influence of fever on biliary elements of guinea pigs

ＩｎｆｌｕｅｎｃｅｏｆｆｅｖｅｒｏｎｂｉｌｉａｒｙｅｌｅｍｅｎｔｓｏｆｇｕｉｎｅａｐｉｇｓＬüＨｏｕＤｏｎｇ１，ＴＩＡＮＭｉｎｇＧｕｏ２，ＺＨＡＮＧＸｉａｏＰｅｎｇ２ａｎｄＬＩＨｕａｉＬａｎ３１ＤｅｐａｒｔｍｅｎｔｏｆＭｉｃｒｏｂｉｏｌｏｇｙ...     

食管癌组织CD15抗原和组织蛋白酶D表达的关系

目的探讨CD15抗原和组织蛋白酶D(Cath-D)在食管癌的表达意义及其相互关系.方法应用微波-LSAB免疫组化法,观察65例食管癌组织(男46例,女19例;平均年龄54岁;均为鳞癌)中CD15和Cath-D的表达阳性率及其相互关系.结果食管癌CD15和CD阳性率分别为58%(38/65例)和65%(42/65例);食管癌CD15和Cath-D表达与性别、年龄、肿瘤部位、大体类型及肿瘤大小无关(P＞0.05).CD15和Cath-D表达阳性率:鳞癌Ⅲ级(88%,14/16 vs 100%,16/16)均显著高于鳞癌Ⅰ级(43%,6/14 vs 50%,7/14)和鳞癌Ⅱ级(51%,18/35 vs 54%,19/35,P＜0.05);外膜层浸润(74%,20/27 vs82%,22/27)显著高于肌层浸润(47%,18/38 vs 53%,20/38,P＜0.05);有淋巴结转移组(72%,26/36 vs 78%,28/38)显著高于无淋巴结转移组(41%,12/29 vs 48%,14/29,P＜0.025);死亡组(70%,31/44 vs 77%,34/44)显著高于五年生存组(33%,7/21 vs 38%,8/21,P＜0.01).CD15阳性的肿瘤Cath-D阳性率显著高于CD15阴性者(82%,31/38 vs 41%,11/27),CD15表达与Cath-D表达呈正相关(r=0.42,P＜0.01).结论检测CD15和Cath-D表达对判断食管癌恶性程度、预测其侵袭转移趋势和预后及指导治疗有重要意义.食管癌CD15与Cath-D表达具有相互协同作用可能.     

Transepithelial transport of putrescine across monolayers of the human intestinal epithelial cell line, Caco-2

AIM: To study the transepithelial transport characteristics of the polyamine putrescine in human intestinal Caco-2 cell monolayers to elucidate the mechanisms of the putrescine intestinal absorption. METHODS: The transepithelial transport and the cellular accumulation of putrescine was measured using Caco-2 cell monolayers grown on permeable filters. RESULTS: Transepithelial transport of putrescine in physiological concentrations (> 0.5 mM) from the apical to basolateral side was linear. Intracellular accumulation of putrescine was higher in confluent than in fully differentiated Caco-2 cells, but still negligible (less than 0.5%) of the overall transport across the monolayers in apical to basolateral direction.EGF enhanced putrescine accumulation in Caco-2 cells by four fold, as well as putrescine conversion to spermidine and spermine by enhancing the activity of S adenosylmethionine decarboxylase. However, EGF did not have any significant influence on putrescine flux across the Caco-2 cell monolayers. Excretion of putrescine from Caco-2 cells into the basolateral medium did not exceed 50 picomoles, while putrescine passive flux from the apical to the basolateral chamber, contributed hundreds of micromoles polyamines to the basolateral chamber. CONCLUSION :Transepithelial transport of putrescine across Caco2 cell monolayers occurs in passive diffusion, and is not influenced when epithelial cells are stimulated to proliferate by a potent mitogen such as EGF.     

Changes of lipid metabolism in plasma, liver and bile during cholesterol gallstone formation in rabbit model

AIM To find out the relationship between the disturbances of lipid metabolism and the formation of cholesterol gallstones by studying the changes of lipid metabolism in plasma, liver tissue and the bile.METHODS Male and female white Japanese rabbits were divided randomly into a control group (Con) and four experimental groups of 10 rabbits each fed with a diet containing 1.2% cholesterol for one, two, three and four weeks (1wk, 2wk, 3wk and 4wk group). The measurement of plasma triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and its subfractions (HDL2-C, HDL3-C), very low and low density lipoprotein cholesterol (VLDL-C, LDL-C) was taken with standard enzymatic techniques. Apolipoprotein (apo) concentrations in plasma were measured by radial immunodiffusion assay for apoA1, apoB100, aopCⅡ and apoCⅢ. Total cholesterol of liver was measured by the enzymatic procedure for each animal. Bile acids, mainly glycocholate (GCA) and glycodeoxycholate (GDCA) were detected by dual wavelength thin layer scanner.RESULTS In all the experimental groups fed with dietary cholesterol, cholesterol crystal was found in the gallbladder in 2/10 cases of the 1wk group, 4/10 of the 2wk group, 6/10 of the 3wk group and 7/10 of the 4wk group respectively. The concentration of plasma total cholesterol (TC), triglyceride (TG), phospholipid (pl), VLDL-C, LDL-C, apoB100, apoCⅡ, apoCⅢ gradually increased (P＜0.05) with the prolonged feeding time of dietary cholesterol. High density lipoprotein cholesterol and its subfractions (HDL-C, HDL2-C, HDL3-C) showed a tendency to decrease, but without statistical significance (P＞ 0.05). ApoA1 was reduced with increased feeding time of dietary cholesterol (P ＜0.05). The hepatic and biliary cholesterol increased 1-1.5 times as compared with the control group (t=5.221 and 3.445, P＜0.05). The GCA gradually decreased beginning from the control group to the 4wk group (P＜0.05).CONCLUSION Owing to the high cholesterol diet, the increased concentrations of plasma TC, TG, VLDL-C, LDL-C, hepatic TC and TG, apoB100, apoCⅡ and apoCⅢ possibly enhanced the secretion of biliary cholesterol into bile; the decreased plasma apoA1 level might reduce the secretion of antinucleating factor into bile. All those factors mentioned above probably contribute to the formation of cholesterol gallstones.     

Effects of hypoxia, hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 in hepatic stellate cells

AIM To study the effects of hypoxia,hyperoxia on theregulation of expression and activity of matrixmetalloproteinase-2 (MMP-2) in hepatic stellate cells(HSC).METHODS The expressions of MMP-2,tissue inhibitor ofmatrix metalloproteinass-2 (TIMP-2) and membrane typematrix matalloproteinass-1 (MT1-MMP) in cultured rat HSCwere detected by immunocytochemistry (ICC) and in situhybridization (ISH).The contents of MMP-2 and TIMP-2 inculture supernatant were detected with ELISA and theactivity of MMP-2 in supernatant was revealed byzymography.RESULTS In the situation of hypoxia for 12h,theexpression of MMP-2 protein was enhanced (hypoxiagroup positive indexes:5.7±2.0,n=10;control:3.2±1.0,n=7;P<0.05),while TIMP-2 protein was decreasedin HSC (hypoxla group positive indexes:2.5±0.7,n=10;control:3.6±1.0,n=7;P<0.05),and the activity(total A) of MMP-2 in suparnatant declined obviously(hypoxla group:7.334±1.922,n=9;control:17.277±7.424,n=11;P<0.01).Compared the varied duration ofhypoxia,the changes of expressions Including mRNA andprotein level as well as activity of MMP-2 were mostnotable in 6 h group.The highest value (Ahypoxla~-Acontrol) ofthe protein and the most intense signal of mRNA were inthe period of hypoxia for 6 h,along with the lowestactivity of MMP-2.In the situation of hyparoxia for 12 h,the contents (A_(450)) of MMP-2 and TIMP-2 in supernatantwere both higher than those in the control,especially theTIMP-2 (hyperoxla group:0.0499±0.0144,n=16;control:0.0219±0.0098,n=14;P<0.01),and so wasthe activity of MMP-2 (hyperoxia group:5.252±0.771,n=14;control:4.304±1.083,n=12;P<0.05),and the expression of MT1-MMP was increased.CONCLUSION HSC is sensitive to the oxygen,hypoxiaenhances the expression of MMP-2 and the effect is moremarked at the early stage;hyperoxia mainly raises theactivity of MMP-2.     

表皮生长因子受体和PCNA在食管癌及癌前病变组织中的表达意义

目的探讨ＥＧＦＲ及ＰＣＮＡ的表达在食管癌发生发展过程中的临床意义．方法采用免疫组化ＳＰ法对各级食管鳞癌（Ⅰ级ｎ＝１８，Ⅱ级ｎ＝３０，Ⅲ级ｎ＝２２）、各级不典型增生（轻度ｎ＝１４，中度ｎ＝１７，重度ｎ＝１３）及正常食管粘膜（ｎ＝２０）进行ＥＧＦＲ及ＰＣＮＡ检测．结果ＥＧＦＲ及ＰＣＮＡ阳性表达在各组食管病变中呈明显递增趋势，正常组、轻度及中度不典型增生组间ＥＧＦＲ及ＰＣＮＡ表达均有显著性差异，ＥＧＦＲ阳性表达分别为：０／２０，３／１４及１０／１７；ＰＣＮＡⅠ，Ⅱ，Ⅲ，Ⅳ级表达分别为：１９／２０，１／２０，０／２０，０／２０；７／１４，６／１４，１／１４，０／１４；２／１７，７／１７，７／１７，１／１７（Ｐ＜００５）．鳞癌Ⅰ，Ⅱ级组间ＰＣＮＡ表达有显著性差异，ＰＣＮＡⅠ，Ⅱ，Ⅲ，Ⅳ级表达分别为１／１８，７／１８，８／１８，２／１８；２／３０，５／３０，８／３０，１５／３０（Ｐ＜００５）；而中、重度不典型增生与Ⅰ级鳞癌组间均无显著性差异（Ｐ＞００５）．结论食管中重度不典型增生者，其基因表达及增殖特性已具有明显的潜在恶性趋势，与食管癌的发生有直接关系，临床应对这类病例积极治疗并密切随访，以防癌变