AQP7和AQP9基因多态性与中国汉族人群2型糖尿病的相关性研究CSCD

目的探讨水通道蛋白7(aquaporin 7, AQP7)以及水通道蛋白9(aquaporin 9, AQP9)基因单核苷酸多态性(single nucleotide polymorphism, SNP)与中国汉族人群患2型糖尿病(type 2 diabetes mellitus, T2DM)的相关性。方法随机纳入1194例T2DM个体和1274例非糖尿病个体(non-diabetic, NDM)进行对照研究, 采用MassArray质谱基因分型方法对3个SNP位点(AQP7基因rs3758269、AQP9基因rs16939881和rs57139208)进行基因分型。评估以上3个SNP位点与T2DM的相关性;探讨NDM组SNP位点处不同基因型与糖脂代谢指标的关联。结果 AQP7基因rs3758269、AQP9基因rs16939881和rs57139208的等位基因频率及基因型频率在T2DM组和NDM组中的分布无统计学差异(P > 0.05);且分析结果显示不同遗传模式与T2DM无相关性(P > 0.05)。在NDM组中, AQP7基因rs3758269、AQP9基因rs16939881和rs57139208的不同基因型与糖脂代谢指标无相关性(P > 0.05)。结论 AQP7基因rs3758269和AQP9基因rs16939881和rs57139208与中国汉族人群T2DM遗传易感性无关。     

CDKAL1基因rs4712523多态性与内蒙古地区汉族人群2型糖尿病相关

目的 研究内蒙古地区汉族人群CDKAL1基因rs4712523单核苷酸多态性(SNP)的等位基因和基因型频率分布与2型糖尿病(T2DM)的相关性.方法 采用等位基因特异性聚合酶链式反应(AS-PCR),对382例内蒙古地区汉族人(其中T2DM组192例,对照组190例)rs4712523进行基因分型.结果 T2DM组中rs4712523的G等位基因频率和GG基因型频率分别为47.4％和6.3％,均显著高于对照组的35.3％和3.2％(P＜0.05).G等位基因携带者患T2DM的风险是A等位基因的1.654倍(OR=1.654,95％ CI=1.237-2.212).结论 CDKAL1基因rs4712523多态性位点的G等位基因可能是内蒙古地区汉族人T2 DM的易感基因之一.     

clock基因rs1801260多态性与抑郁症的关联分析

目的:探讨clock基因与汉族人群抑郁症的关联。方法:用Snapshot SNP分型技术对155例抑郁症患者(其中包括133名抑郁症睡眠障碍者)和150名正常对照进行clock基因rs1801260位点分型,比较两组该基因多态性基因型和等位基因频率的差异。结果:与对照组相比,患者组rs1801260多态性的基因型和等位基因的频率差异无统计学意义(P0.05);对rs1801260多态性二种基因型C/T、T/T抑郁症的临床资料比较,显示HAMA总分差异具有统计学意义(t=2.012,P=0.047),其他各项没有明显差异(P0.05)。结论:clock基因rs1801260多态性可能与中国汉族抑郁症的发病无关联,但与抑郁症的焦虑程度可能关联。     

云南汉族心肌梗死相关基因的多态性研究

目的 探索凝血因子Ⅶ(coagulation factor Ⅶ,FⅦ)、凝血因子Ⅻ(coagulation factor Ⅻ,FⅫ)、人血纤蛋白原( fibrinogen,FBG)及9p21的基因多态在云南汉族人群中的分布及其与心肌梗死的相关性.方法 应用聚合酶链反应-限制性片段长度多态和焦磷酸测序分型技术对142例心肌梗死患者和192名对照者的FⅦ、FⅫ、FBG、9p21多个位点的基因进行检测分析.结果 R353Q、5′F7、C46T、-148C/T、rs1333049、rs4977574位点的多态性在心肌梗死组与对照组之间差异无统计学意义(P＞0.05),-455G/A的AA基因型、rs1333040的T等位基因和TT基因型、rs10116277的T等位基因和TT基因型、rs2383207的G等位基因和GG基因型频率均显著高于对照组(P＜0.05),而rs1333040的CT和rs10116277的GT基因型频率均显著低于对照组(P＜0.05).结论 FⅦ、FⅫ、FBG基因的-148C/T、9p21的rs1333049位点的多态性与云南汉族心肌梗死无显著关联,FBG基因的-455G/A和9p21的rs1333040、rs10116277、rs2383207位点的多态性可能与云南汉族心肌梗死有一定的相关性.     

云南地区汉族人群PSMB8、PSMB9及TAP2基因多态性与类风湿关节炎的关联研究

目的 研究云南汉族人群中PSMB8、PSMB9及TAP2基因多态性与类风湿关节炎(rheumatoid arthritis,RA)的相关性.方法 应用聚合酶链反应-限制性片段长度多态性法对177例RA患者及288名健康对照PSMB8基因的rs2071543、rs55745125、rs138635403位点和PSMB9基因的rs17587多态性位点进行基因分型,应用多聚酶链反应扩增阻碍系统法对TAP2基因的rs2228396多态性位点进行基因分型.计算基因型及等位基因频率.采用Epi Info 7软件计算上述多态位点在RA组及正常对照组之间的比值比(OR值).结果 rs138635403及rs17587位点的等位基因及基因型频率在RA组和对照组间差异有统计学意义(P＜0.05),其中RA组rs17587的GG基因型频率(0.672)高于对照组(0.524)(OR=1.862,95％CI:1.261～2.749).结论 云南汉族人群PSMB9基因的rs17587位点多态性与RA存在关联.     

TLR7基因单核苷酸多态性与江苏淮安哮喘患者易感性的相关性研究

目的:探讨Toll样受体7(TLR7)基因rs179009、rs179019、rs5935436位点多态性与江苏淮安地区汉族人群支气管哮喘的相关性。方法:采用病例对照方法,以聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法比较158例支气管哮喘组与137例健康对照组之间基因型、等位基因频率的差异。结果:哮喘组TLR7基因rs179009、rs170019位点基因型及等位基因型频率与对照组相比差异有统计学意义(P<0.05)。rs5935436位点基因型及等位基因型频率在哮喘组和对照组间差异均无统计学意义(P>0.05)。结论:TLR7基因rs179009位点和rs179019位点的多态性可能与江苏淮安地区汉族人群哮喘相关;而rs5935436位点的多态性可能与江苏淮安地区汉族人群哮喘无关。     

糖皮质激素受体遗传多态性与2型糖尿病相关性研究

目的 探讨糖皮质激素受体(GR)单核苷酸多态性与2型糖尿病(T2MD)易感性及人体体型的相关性.方法 采用病例-对照研究设计,从解放军总医院健康体检中心的体检患者中,募集40例T2MD患者和127例对照.结合三种不同的SNP位点选择方法,采用上海天昊生物科技有限公司的iMLDRTM多重SNP分型技术对入选SNP位点进行基因分型.采用非条件Logistic回归,校正年龄、性别、吸烟、饮酒后,分析基因型与T2MD易感性的关系.得到的阳性位点,进一步采用协方差分析,校正年龄、性别、吸烟、饮酒后,评价其与体重指数(BMI)和腰臀比(WHR)的相关性.结果 共选取14个SNP位点,其中rs10052957突变型在病例组中频率为零,故未纳入统计分析.所有的13个SNP位点在病例和对照组中的基因分型均符合Hardy-Weinberg平衡.rs9324924 TT基因型(OR [95％CI]=3.12[1.06 ～9.17],P=0.039)和rs9324921 AA基因型(OR [95％CI]=14.92[1.39～160.60],P=0.026)发生T2DM的风险较野生型增高,且两基因型的BMI[P=0.023 (rs9324924);P=0.002(rs9324921)]、WHR[P=0.033 (rs9324924);P=0.003(rs9324921)]也较野生型明显增高.结论 在本研究中,GR基因rs9324924、rs9324921与T2DM易感性和体型有关,其中纯和突变型T2DM发病风险明显增加,且更具有肥胖倾向.     

VARS2基因单核苷酸多态性和广东汉族女性散发性乳腺癌相关性研究

目的探讨VARS2(valyl-tRNA synthetase 2)基因中单核苷酸多态性位点(single nucleotide polymorphisms,SNP)rs2249464与广东省汉族女性散发性乳腺癌的相关性。方法采用MassARRAY-IPLEX SNP分型技术,以南方医科大学南方医院的216例广东汉族乳腺癌患者及216例同期健康体检者为研究对象,对rs2249464多态性位点进行基因分型,利用χ2检验统计分析病例组和对照组的基因型频率有无差异,利用非条件Logistic回归计算比数比(odds ratio,OR)和95%可信区间(confidence interval,CI)来评价此位点多态性与乳腺癌的相关性。然后进一步将病例组按雌激素受体(estrogen receptor,ER)和孕激素受体(progesterone receptor,PR)免疫组化结果行分层分析。结果 rs2249464位点C/C,C/T,T/T 3种基因型分布在对照组和病例组之间有统计学差异。根据免疫组化情况进一步分层分析结果表明rs2249464的基因型分布在ER阳性/阴性组之间的差异无统计学意义(P>0.05);但在PR阳性/阴性组之间的差异有统计学意义(P<0.05)。结论 VARS2基因rs2249464位点单核苷酸多态性与广东汉族散发性乳腺癌的易感性具有相关性,且携带T/T基因型的患者更容易罹患PR阴性乳腺癌。     

Kir4.1和AQP4基因多态性与颞叶癫痫遗传易感性的关系

 目的 探讨在星形胶质细胞中高表达的两个基因Kir4.1和AQP4多态性与颞叶癫痫遗传易感性的关系。方法 采用病例-对照研究策略,以中国汉族人群438例颞叶癫痫患者和695例非癫痫受试者为研究对象,应用TaqMan法检测Kir4.1和AQP4基因的多态性.。结果 共选择Kir4.1基因上的5个SNP位点以及AQP4基因上的7个SNP位点,在病例组和对照组的基因型分布均符合Hardy-Weinberg平衡,这些多态性位点与颞叶癫痫之间并无相关性,基因单倍型与疾病的关联无显著性的差异,结合临床表型进行分层分析,亦未有显著差异。结论 在中国汉族人群中,Kir4.1和AQP4基因多态性在颞叶癫痫的发生和发展过程中无重要作用。     

CSPG2和HSPG2基因单核苷酸多态性与中国汉族人颅内动脉瘤的相关性研究

目的 分析CSPG2和HSPG2基因单核苷酸多态性(single nucleotide polymorphism,SNP)与中国汉族散发颅内动脉瘤的相关性.方法 采用病例-对照关联研究方法,收集颅内动脉瘤患者537例以及年龄和性别匹配的正常对照1071名的外周血样各5 mL并提取基因DNA.通过聚合酶链式反应扩增目的DNA,用单碱基延伸(SNaPshot)法进行SNP分型.选取文献报道的CSPG2和HSPG2基因的两个标签SNPs位点rs251124和rs3767137,分析其与汉族散发颅内动脉瘤发病的关联性.结果 CSPG2和HSPG2基因的两个标签SNPs位点rs251124和rs3767137的基因型均满足Hardy-Weinberg平衡.CSPG2rs251124的等位基因频率在患者组与对照组之间差异无统计学意义(P=0.22);HSPG2 rs3767137的等位基因频率在两组之间亦差异无统计学意义(P=0.26),但其相应的OR值大于1(OR=1.12;95％CI=0.92～1.37).患者组与对照组rs251124、rs3767137的基因型频率均差异无统计学意义(P=0.46,0.53).结论 未发现CSPG2和HSPG2基因rs251124、rs3767137 SNPs与中国人颅内动脉瘤发病的相关性.     

Immunohistochemical Localization of the Aquaporins AQP1, AQP3, AQP4, and AQP5 in the Mouse Respiratory System

Aquaporins are membrane water channel proteins that function mainly in water transfer across cellular membranes. In our present study, we investigated the immunohistochemical distribution of aquaporin 1 (AQP1), AQP3, AQP4, and AQP5 in the mouse respiratory system by immunofluorescence, immunoperoxidase, and immunoelectron microscopy. AQP3, AQP4, and AQP5 are expressed in epithelial cells, whereas AQP1 is expressed in subepithelial connective tissues and capillaries. In the airway surface epithelia from the nasal cavity to the intrapulmonary bronchioles, AQP5 was found to be mainly localized to the luminal side and both AQP3 and AQP4 to the abluminal side. In the alveolar epithelium, AQP5 is localized to the apical membranes of both type I and type II alveolar cells. Compared with the previous studies on the rat respiratory system, in which AQP5 is restricted to the alveolar type I cells and absent from the airway surface epithelia, we found that AQP5 in the mouse is much more widely distributed throughout the surface epithelia. These results suggest that AQP5 has a critical role in water-handling, such as the maintenance of airway surface liquid and clearance of alveolar fluid in the mouse respiratory system.     

激活的人外周T淋巴细胞表达水通道蛋白AQP3

目的:探讨水通道蛋白(Aquaporin,AQP)在人外周T淋巴细胞的表达和生理学意义.方法:利用逆转录PCR和免疫印迹技术检测AQP0～9 mRNA和蛋白在静止的和经活化的T淋巴细胞的表达.结果:静止的淋巴细胞无AQP0-9表达,而活化的T淋巴细胞表达AQP3 mRNA和蛋白.结论:T淋巴细胞激活过程可能需要AQP3转运水和甘油的功能.     

尼莫同对严重烫伤幼年大鼠脑内水通道蛋白4表达的影响

目的探讨幼年大鼠严重烫伤后脑内水通道蛋白4（AQP4）的表达和脑含水量变化的影响。方法75只健康Wistar幼年大鼠随机分为烫伤组、尼莫同组和假烫伤组，每组25只。建立严重烫伤模型，所有实验大鼠分别在烫伤后2h、6h、12h、24h、48h断头处死后取脑．测定脑含水量，并用免疫组化测定AQP4的表达。结果烫伤组脑含水量及AQP4表达在2h后开始增高，6h后达高峰，尼莫同组与烫伤组和假烫伤组比较，各时间点脑含水量明显降低（P〈0．05）。结论AQP4与脑水肿形成有关，尼莫同可以降低脑组织内AQP4的表达．从而减轻脑水肿。     

Further evidence for AQP8 expression in the myoepithelium of rat submandibular and parotid glands

Previously (Wellner et al., Pflugers Arch 441:49–56, 2000) we suggested that the localization of the aquaporins (AQPs) AQP5 and AQP8 in the apical and basolateral membranes of rat submandibular gland (SMG) acinar cells, respectively, provides for transcellular water flow during saliva formation. While the localization of AQP5 in this gland has been verified in several laboratories, there have been differing reports regarding AQP8 localization. Other investigators subsequently reported that AQP8 is not expressed in the acinar or ductal cells of the major salivary glands of the rat, but in the myoepithelium of each gland. Thus, we have carried out additional studies: (1) to reassess the localization of AQP8 in the rat SMG and (2) to assess the localization of AQP8 in the rat parotid gland (PG). Initially, we compared the localizations of AQP8 with recognized basolateral markers in acinar cells [the Na⁺,K⁺-ATPase and the Na⁺–K⁺–2Cl^– cotransporter (NKCC1)]. Our results indicated that Na⁺,K⁺-ATPase localized in both the basal and lateral membranes of rat SMG acinar cells, whereas AQP8 was detected only in the basal regions of the acini. In the rat PG, AQP8 was invested near intercalated ducts and adjacent acini, whereas NKCC1 localized in the basolateral membranes of acinar cells. As these results were suggestive of myoepithelial localization in both glands, we compared AQP8 localization with the localization of smooth muscle actin, a myoepithelial marker. We found that AQP8 and smooth muscle actin colocalized in both the rat SMG and PG, providing additional strong support for a myoepithelial localization of AQP8. Thus, in agreement with an earlier report by other investigators (Elkjaer et al., Am J Physiol Renal Physiol 281:F1047–F1057, 2001), we report that AQP8 is expressed in the myoepithelial cells, but not in the acinar cells, of both the rat SMG and PG.     

食管胃连接部癌中Ezrin、AQP5的表达

目的观察Ezrin、AQP5蛋白在冀南地区食管胃连接部癌中的表达和相关性,探讨其生物学行为。方法应用免疫组化Max Vision法检测10例正常胃组织、40例异型增生胃黏膜组织和165例食管胃连接部癌组织中Ezrin、AQP5蛋白的表达。结果Ezrin、AQP5蛋白在正常胃组织、异型增生胃黏膜组织和食管胃连接部癌组织中的阳性率依次增高,分别为10%、45%、72.7%和20%、32.5%、65.5%。二者在三组间相比差异均有显著性(P<0.05)。Ezrin、AQP5蛋白在分化程度低、侵及浆膜层及有淋巴结转移的食管胃连接部癌中的表达均明显高于分化程度高、未侵及浆膜层以及无淋巴结转移组(P<0.01)。Ezrin与AQP5在食管胃连接部癌中表达呈正相关性(P<0.05)。结论 Ezrin和AQP5在食管胃连接部癌中均高表达,且呈正相关关系,二者可能起协同作用,共同参与食管胃连接部癌的侵袭和转移。     

Functional involvement of Annexin-2 in cAMP induced AQP2 trafficking

Annexin-2 is required for the apical transport in epithelial cells. In this study, we investigated the involvement of annexin-2 in cAMP-induced aquaporin-2 (AQP2) translocation to the apical membrane in renal cells. We found that the cAMP-elevating agent forskolin increased annexin-2 abundance in the plasma membrane enriched fraction with a parallel decrease in the soluble fraction. Interestingly, forskolin stimulation resulted in annexin-2 enrichment in lipid rafts, suggesting that hormonal stimulation might be responsible for a new configuration of membrane interacting proteins involved in the fusion of AQP2 vesicles to the apical plasma membrane. To investigate the functional involvement of annexin-2 in AQP2 exocytosis, the fusion process between purified AQP2 membrane vesicles and plasma membranes was reconstructed in vitro and monitored by a fluorescence assay. An N-terminal peptide that comprises 14 residues of annexin-2 and that includes the binding site for the calcium binding protein p11 strongly inhibited the fusion process. Preincubation of cells with this annexin-2 peptide also failed to increase the osmotic water permeability in the presence of forskolin in intact cells. Altogether, these data demonstrate that annexin-2 is required for cAMP-induced AQP2 exocytosis in renal cells.     

水通道蛋白4在大鼠眼球中的表达

目的从转录和翻译水平观察水通道蛋白4(AQP4),在正常Wistar大鼠眼球中的表达,为研究眼球中液体转运机制提供形态学基础。方法采用免疫组织化学SABC法和原位杂交技术。结果所测大鼠眼球呈较强的AQP4免疫反应阳性,角膜上皮细胞、虹膜色素上皮细胞、睫状体非色素上皮细胞、晶状体上皮细胞、晶状体纤维细胞、视网膜的内颗粒层、外颗粒层和节细胞层的细胞均为阳性细胞,免疫反应阳性物质主要分布在细胞膜上,胞核呈阴性。原位杂交结果同免疫组织化学染色基本一致,在以上细胞胞浆中均可检测到较强的AQP4 mRNA阳性杂交信号,‘胞核呈阴性。结论AQP4可能在眼球中角膜和晶状体透明性的维持,房水的分泌和调节,视觉的激发和传递等功能活动中起着重要作用。     

水通道蛋白-4在大鼠大脑中的定位研究

目的 观察水通道蛋白-4(AQP4)在大脑实质和室管膜、脉络丛、最后区、神经垂体、腺垂体和松果体中的分布,为研究脑组织中水分子运输和平衡以及内分泌腺体的分泌和调节机制提供形态学基础。方法 免疫组织化学漂洗法(LAB-SA、BCIP／NBT法)。结果 AQP4主要分布于软脑膜、脑室和导水管系统的室管膜、脉络丛等与脑脊液(CSF)直接接触的组织中及脑实质的血管周围;在脑实质内可见阳性细胞体;在海马的锥体细胞层、齿状回的颗粒细胞层和多形细胞层细胞呈阳性;在脑室周围器官如神经垂体、最后区(AP)及松果体和腺垂体(包括中间部)各种腺细胞膜表面均可见AQP4的阳性标记。结论 AQP4不仅与水分子的转运及平衡调节有关、同时与电解质代谢和内分泌活动有关,还可能与下丘脑的神经内分泌活动有关。     

Upregulated Expression of AQP 7 in the Skeletal Muscles of Obese ob/ob Mice

Aquaporin (AQP) is suggested to be regulated by leptin through the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin pathway. AQP7 and AQP9 are membrane proteins with water and glycerol channels, the latter of which is essential for triglyceride synthesis. We conjectured that the expression of AQP7 and AQP9 would be altered in the skeletal myofibers in obese leptin deficient ob/ob mice as compared with that of wild mice. RNA and protein levels were studied in the quadriceps femoris muscles of ob/ob and wild mice. Real time quantitative RT-PCR analysis showed that mouse AQP7 mRNA levels in skeletal muscles were significantly higher in ob/ob mice than in wild mice (P<0.01), whereas mouse AQP9 mRNA level was not different between the two groups (P>0.05). Histologically the type 1 myofibers of ob/ob mice contained numerous lipid droplets in oil red O stain samples. Immunohistochemical staining of ob/ob mouse muscles revealed enhanced expression of AQP7 at myofiber surface membranes, while AQP9 expression appeared to be similar to that of wild mice. The findings suggest that the upregulated expression of AQP7 in ob/ob mouse muscles facilitates the secretion of glycerol from myocytes.