CN111710435A - 血浆维生素d水平与食管鳞癌发生的关系的研究方法 - Google Patents
血浆维生素d水平与食管鳞癌发生的关系的研究方法 Download PDFInfo
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Abstract
本发明适用于食管鳞癌研究的技术领域,提供了血浆维生素D水平与食管鳞癌发生的关系的研究方法,首先通过收集不同地区的患者、健康和健康的移民人群作为研究组,然后采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平,接着进行统计学处理,最后比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断,本发明操作简单,可以为血浆维生素D水平与食管鳞癌发生的关系提供有效的理论依据。
Description
技术领域
本发明属于食管鳞癌研究的技术领域,尤其涉及血浆维生素D水平与食管鳞癌发生的关系的研究方法。
背景技术
维生素D缺乏和多种慢性代谢性疾病包括肿瘤、自身免疫病、糖尿病、心血管疾病等密切相关;多项研究还表明维生素D对肿瘤患者预后有重要影响。2J。河南林州(原林县)是世界食管癌高发区之一"J,其北部地区部分居民113年前移居到山西长治地区,目前形成了较为集中的自然移民村,他们中98%以上的居民是林州人后裔,生活环境、饮食习惯已与林州地区发生了较大变化,而与山西长治当地情况相似。本研究利用ELISA法检测河南林州食管鳞癌患者、河南林州健康人群、山西长治健康人群、山西长治食管鳞癌患者、长治林州移民健康人群这5组人群血浆中维生素D的水平,旨在探讨血浆维生素D水平与食管鳞癌发生的关系。
发明内容
本发明提供血浆维生素D水平与食管鳞癌发生的关系的研究方法,旨在解决为食管鳞癌的发生因素的研究提供跟多的理论依据。
本发明是这样实现的,血浆维生素D水平与食管鳞癌发生的关系的研究方法,包括以下步骤:
S1、收集a地的预设数量的食管鳞癌患者作为研究组A1;收集a地的预设数量的无食管、贲门及胃黏膜癌变者作为研究组A2;收集b地的预设数量的无食管、贲门及胃黏膜癌变者作为研究组B1;收集b地的预设数量的食管鳞癌患者作为研究组B2;收集b地的预设数量的无食管、贲门及胃黏膜癌变者的a地移民后裔作为研究组C;
S2、采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平;
S3、采用ELISA法检测5组研究组人群的血浆维生素D的水平;
S5、比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断。
优选的,所述步骤S2中,维生素D的摄入量来源于膳食和营养补充剂。
优选的,所述步骤S3中,采用ELISA法检测5组研究组人群的血浆维生素D的水平,具体为:
样本处理:以柠檬酸钠或乙二胺四乙酸作为抗凝剂,混合10min,2000r/min离心15min,收集上清,在-20℃的温度条件下冻存待测;
操作步骤:按照人维生素D酶联免疫检测试剂盒说明书进行;
计算维生素D水平:依据标准品的浓度及对应的吸光度值计算样品浓度。
与现有技术相比,本发明的有益效果是:本发明的血浆维生素D水平与食管鳞癌发生的关系的研究方法,首先通过收集不同地区的患者、健康和健康的移民人群作为研究组,然后采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平,接着进行统计学处理,最后比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断,本发明操作简单,可以为血浆维生素D水平与食管鳞癌发生的关系提供有效的理论依据。
附图说明
图1为本发明的的流程示意图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合附图及实施例,对本发明进行进一步详细说明。应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。
请参阅图1,本发明提供一种技术方案:血浆维生素D水平与食管鳞癌发生的关系的研究方法,包括以下步骤:
S1、将研究对象分为5组:
A1组:河南林州食管鳞癌患者:收集2012年6月-2014年5月在长治医学院附属和平医院中心医院进行胃镜检查并经病理证实的食管鳞癌患者104例,男34例,女70例,年龄5l一70岁;
A2组:河南林州健康人群:收集同期在长治医学院附属和平医院中心医院进行胃镜检查,证实无食管、贲门及胃黏膜癌变者104例,男56例,女48例,年龄48-72岁。
B1组:山西长治健康人群:收集2013年9月-2014年5月在长治医学院附属和平医院中心医院进行胃镜检查,证实无食管、贲门及胃黏膜癌变者104例,男60例,女44例,年龄43~78岁。
B2组:山西长治食管鳞癌患者:收集同期在长治医学院附属和平医院中心医院进行胃镜检查,病理证实的食管鳞癌患者108例,男48例,女60例,年龄45~75岁。
C组:长治地区的健康林州移民人群:收集2011年6月-2013年9月在长治医学院附属和平医院中心医院进行胃镜检查,证实无食管、贲门及胃黏膜癌变的林州移民后裔80例,男26例,女54例,年龄44~77岁。
如下表1所示:
S2、采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平。维生素D的摄入量来源于膳食和营养补充剂。调查问卷显示:以上5组人群饮食摄人的维生素D基本一致,均无含维生素D药物的补充。
S3、采用ELISA法检测5组研究组人群的血浆维生素D的水平。
样本处理:以柠檬酸钠或乙二胺四乙酸作为抗凝剂,混合10min,2000r/min离心15min,收集上清,在-20℃的温度条件下冻存待测;
操作步骤:按照人维生素D酶联免疫检测试剂盒说明书进行;
计算维生素D水平:依据标准品的浓度及对应的吸光度值计算样品浓度。
其中,括号内例数。
S5、比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断。
在本实施例中,结果如下:
5组的总体血浆维生素D水平及不同性别、年龄的血浆维生素D水平的差异均有统计学意义(P均<0.05)。用Bonfferoni法对5组的血浆维生素D水平进行两两比较:
总体水平比较:林州健康组与林州食管癌组比较,差异有统计学意义(P<0.05);林州食管癌组与林州移民健康组比较,差异有统计学意义(P<0.05)。长治食管癌组与林州健康组比较,差异有统计学意义(P<0.05);长治食管癌组与林州移民组比较,差异有统计学意义(P<0.05)。
女性人群:林州健康组与林州食管癌组比较,差异有统计学意义(P<0.05);林州食管癌组与林州移民组比较,差异有统计学意义(P<0.05)。
大于60岁人群:林州健康组与林州食管癌组比较,差异有统计学意义(P<0.05);林州食管癌组与林州移民组比较,差异有统计学意义(P<0.05);长治食管癌组与林州健康组比较,差异有统计学意义(P<0.05);长治食管癌组与林州移民组比较,差异有统计学意义(P<0.05)。
食管癌的发生与多种环境、营养和基因因素及其交互作用密切相关。迄今为止,已发现与食管癌发生风险显著相关的因素包括年龄、家族史、膳食因素等H1。我们可以通过对相关的饮食和生活习惯等因素进行有效的干预,达到降低食管癌发生风险的目的。在食管癌的发生发展中,营养元素和微量元素缺乏、亚硝酸盐、病毒和一些毒素等被认为是重要的致病因素。以往的调查研究发现,营养缺乏是食管癌高发区较为普遍的现象,在食管癌高发区,维生素D缺乏现象更加明显。近年来的人群研究证据显示,维生素D的缺乏和多种慢性代谢性疾病包括肿瘤、自身免疫病、糖尿病、心血管疾病等密切相关。早在1937年,Peller和Stephenson就发现较长时间暴露于阳光下的人群肿瘤发生率较低。在1941年,Apperly论证了人们居住区域的纬度高低和肿瘤死亡率之间存在负相关性。Garland及同事发现高纬度居住的人们由于体内维生素D的缺乏导致了乳腺癌、结直肠癌、卵巢癌的发生率的上升;Schwartz和Hulka发现维生素D的缺乏同胰腺癌的发生相关。
在通过对肺癌和直肠癌患者的随访研究发现,体内维生素D水平低下者,其总的生存率和生存期限显著低于维生素D处于正常和较高水平的患者。因此,维生素D缺乏是否和食管癌发生相关及具体机制是值得我们探讨的问题。
山西长治地区的林州移民这一人群既有着世界上食管癌发病率最高地区林州人群的遗传背景,同时又有着与长治地区本地居民相似的生活环境和饮食习惯。本研究结果显示:林州健康组与林州食管癌组、林州食管癌组与林州移民组、长治食管癌组与林州健康组、长治食管癌组与林州移民健康组之间的血浆维生素D水平存在差异;林州食管癌组血浆中维生素D水平显著低于林州健康组和林州移民健康组,长治食管癌患者血浆中维生素D水平显著低于林州移民健康组,这表明在食管鳞癌患者中存在维生素D缺乏的现象。同时,男女及年龄分层分析结果显示,林州健康组与林州食管癌组血浆中维生素D水平也具有明显差异。由于长治健康组、长治食管癌组以及林州移民健康组这3组人群的生活环境和饮食习惯相似,我们认为可以排除由于膳食维生素D摄入不同所致的血浆中维生素D水平差异。前期我们发现,在食管鳞癌患者中存在核黄素缺乏的现象。核黄素转运蛋白基因C20orf54若发生变异会导致核黄素不能有效进入细胞内发挥正常作用,我们针对C20orf54基因编码区内单核苷酸多态性(cSNP)位点的研究结果发现,rs3746802、rs3746803和rs3746804位点与食管鳞癌的遗传易感性明显相关。这在一定程度上揭示了环境和遗传因素交互作用对食管癌发生的影响。维生素D在体内以具有生物学活性的1,25(OH)2D3形式与维生素D受体结合,执行生物学功能。综合前期和目前的研究结果,林州移民人群虽然与长治食管癌人群生活环境与饮食习惯相似,但遗传背景不同,如果维生素D受体基因发生变异,将影响机体对维生素D的吸收,从而导致体内维生素D水平下降。维生素D受体的cSNP不同是否是造成体内维生素D水平差异的原因还有待进一步研究
本发明的血浆维生素D水平与食管鳞癌发生的关系的研究方法,首先通过收集不同地区的患者、健康和健康的移民人群作为研究组,然后采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平,接着进行统计学处理,最后比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断,本发明操作简单,可以为血浆维生素D水平与食管鳞癌发生的关系提供有效的理论依据。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (3)
1.血浆维生素D水平与食管鳞癌发生的关系的研究方法,其特征在于:包括以下步骤:
S1、收集a地的预设数量的食管鳞癌患者作为研究组A1;收集a地的预设数量的无食管、贲门及胃黏膜癌变者作为研究组A2;收集b地的预设数量的无食管、贲门及胃黏膜癌变者作为研究组B1;收集b地的预设数量的食管鳞癌患者作为研究组B2;收集a地的预设数量的无食管、贲门及胃黏膜癌变者的b地移民后裔作为研究组C;
S2、采用食物频率调查问卷法对所有研究对象进行膳食调查,评估维生素D摄入水平;
S3、采用ELISA法检测5组研究组人群的血浆维生素D的水平;
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S4、统计学处理:应用SPSSl9.0统计学软件进行数据处理;计量资料以x±s表示,5组间均数的比较采用方差分析,组间两两比较采用Bonfferoni法进行;其中,P<0.05认为差异有统计学意义;
S5、比较各组维生素D水平,进行血浆维生素D水平与食管鳞癌发生的关系的研究判断。
2.如权利要求1所述的血浆维生素D水平与食管鳞癌发生的关系的研究方法,其特征在于:所述步骤S2中,维生素D的摄入量来源于膳食和营养补充剂。
3.如权利要求1所述的血浆维生素D水平与食管鳞癌发生的关系的研究方法,其特征在于:所述步骤S3中,采用ELISA法检测5组研究组人群的血浆维生素D的水平,具体为:
样本处理:以柠檬酸钠或乙二胺四乙酸作为抗凝剂,混合10min,2000r/min离心15min,收集上清,在-20℃的温度条件下冻存待测;
操作步骤:按照人维生素D酶联免疫检测试剂盒说明书进行;
计算维生素D水平:依据标准品的浓度及对应的吸光度值计算样品浓度。
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