目的:观察荷瘤小鼠中期阶段阳气虚证胸腺独特上调和下调的高表达基因.方法:综合采用荷瘤小鼠及其标准化四诊及辨证方法,以及GeneChip Mouse Exon 1.0 ST Array等技术,检测在肿瘤中期阳气虚证H22荷瘤小鼠胸腺组织基因的表达,筛选独特上调或下调者,并重点关注表达量高的基因.结果:肿瘤中期阳虚证荷瘤小鼠胸腺独特上调的基因达260个,下调仅15个.结论:(1)荷瘤小鼠发展到中期的阳气虚证,其胸腺虽然已出现萎缩的现象,但仍处于代偿阶段;(2)Hey2、Hivep2、Stat6、Prdm1、Avpr1a、Ralb、Trdn、Mcmdc1、Olfr1351、Avil、Gns、Ros1等高表达上调的基因,可能与促进T细胞的分化成熟有关;(3)St8sia4、Nxph4、Spred2、Bcas3、Tbcd、Ctns等独特下调基因标志着胸腺内基因表达调节在整体上出现了紊乱,可能预示着胸腺衰竭阶段的到来.这些构成了肿瘤中期阶段阳气虚证荷瘤小鼠胸腺基因表达层面的特征.

Objective: To investigate the characteristics of coincident up- or down-regulated high expressed genes in thymus of H22 tumor-bearing mice of different syndromes.Methods: The thymus gene expression in H22 tumor-bearing mice of pathogenic toxin-exuberance syndrome and qi deficiency syndrome on early stage, yang-qi deficiency syndrome on intermediate stage, qi-yin-yang deficiency syndrome on advanced stage were detected with quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array to screen out the up- and down-regulated genes in uniform, and payed close attention to the highly expressed genes. Results: There are thirty-four gene expression changed obviously after the tumor happened, twenty up-regulated and nine down-regulated. Conclusion: (1)Gene Expression of Tpp2, per2 almost closed on early stage after the tumor happened, they have a close relationship with the early stage of the tumor.(2)The twenty up-regulated genes, such as Hey2, MMP-19, may play a special role during the thymus involution on advanced stage.(3)We think these up- regulated or down-regulated genes may be related to tumorigenesis and its different stage, but not the symptoms and signs.

Objective: To reveal the difference of genes expression in pituitary of H22 tumour mice with sthenia syndrome and asthenia syndrome.Methods: By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array,we observed pituitary gene expression about poisonous pathogenic factors syndrome and asthenia of qi syndrome on earlier period,asthenia of yang and qi syndrome on intermediate stage,asthenia of qi and yin and yang syndrome on advanced stage with H22 tumour mice.Genes expressed highly and differentially distinguished are analyzed on the paper.Screen up-regulated coincident and down-regulated coincident genes in poisonous pathogenic factors syndrome compared to other three syndromes.Results: 121 up-regulated coincident genes were obtained,9 genes whose array indication exceeded 1500 in poisonous pathogenic factors syndrome were introduced: Gabra6,D0H4S114,Alas2,Pvalb,Plp1,Itpr1,Ndrg2,LOC627016,Gpm6b;then 340 down-regulated coincident genes were obtained,21 genes whose array indication exceeded 1500 in normal mice: Abpg,Pip,Dlk1,Car6,Eif3s5,Gnb2,Pcsk1n,Aplp1,Slc22a17,Dnpep,Rnf187,LOC639100,Rab1b,Ald-h2,Pten,Tapbp,2310044H10Rik,Cxxc5,Gatad1,Ren1,Lrp11.Conclusion: After the tumorigenesis,gene expression change is distinguished in pituitary of H22 tumour mice with sthenia syndrome and asthenia syndrome.Some are possible the marker genes of sthenia syndrome and asthenia syndrome.

Objective To investigate genes expression profile in tumor tissues of H22 mice with obstruction of evil syndromes.Methods Genes expression in the tumor tissues of H22 mice with syndromes of obstruction of evil syndromes,"Qi" deficiency,deficiency of "Qi and Yang",or deficiency of "Qi Yin and Yang",were analyzed by Affymetrix GeneChip Mouse Exon 1.0 ST Array.Results 32 special higher-expression genes were observed,including 22 genes which involve in testicle growth,spermatogenesis or oogenesis,4 genes related to tumors,and 6 other gene.Simultaneously,2 special lower-expression genes were observed.Conclusion Numerous genes have special expression in the tumor tissue of H22 mice with obstruction of evil syndromes,which may be intrinsic difference between this syndrome and 3 other syndromes.

目的 揭示H22荷瘤小鼠早期邪毒壅盛证和气虚证肾上腺基因表达的特征.方法 采用小鼠标准化四诊及辨证方法 ,以及GeneChip Mouse Exon 1.0 ST Array等技术,检测H22荷瘤小鼠早期邪毒壅盛证和气虚证、中期阳气虚证、中晚期气阴阳虚证共4个证候肾上腺基因表达的差异,重点关注早期2个证候表达量大、差异显著的基因.结果 ①筛选邪毒壅盛证上调和下调基因20条和7条,气虚证上调和下调基因4条和7条.②邪毒壅盛证上调的基因涉及到信号转导与免疫应答的Pomc1、Tnfaip6、Ccrl2、Rgs1等,调控转录的Bhlhb2、Ell2、Atf3、Hivep2、Klf4等,参与代谢调节的Usp37、Chka、Dusp10等;邪毒壅盛证下调基因有Hbxip、H2-T22、Ndufs6,Tgfb1、Mgat4b、Cdkl3等.③气虚证独特上调基因有Pde10a、Ol-fr1305、Olfr46、Cyp3a13等;气虚证独特下调基因涉及到激素类Gh、Pomc1、Prl、Cga和原癌基因Junb.结论 荷瘤小鼠邪毒壅盛证和气虚证肾上腺存在大量基因表达的改变,这可能是证候内在的本质之一.

[Objective]To reveal the character of neuropeptide gene expression in adrenal gland of H22 tumour mice with different syndromes and signs.[Methods]By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array,we observed hypothalamus and pituitary gland and adrenal gland gene expression about poisonous pathogenic factors syndrome and asthenia of qi syndrome on earlier period,asthenia of yang and qi syndrome on intermediate stage,asthenia of qi and yin and yang syndrome on advanced stage with H22 tumour mice.Forty neuropeptide genes expressed highly and differentially distinguished were analyzed.[Results](1) Fortys neuropeptide genes expression models were different in adrenal gland,for example,GH,PRL,SM,NPY,CCK,POMC and ENK were highly expressed.PACAP,FSH,LH,TSH,SS,GHRH,NT,GLP,PP,CLIP,OT,ADM,SP,SK,DSIP,Gal,AT-Ⅱ and ATG were lowly expressed.However,GnRH,PL,TRH,CRH,VIP,GRP,NPK,DYN,AVP,CGRP,IAPP,UⅡ,ET,BNP and ANP were hardly expressed.(2) Compared with normal mice,down-regulated genes were more than up-regulated genes.LH and ET were up-regulated in poisonous pathogenic factors syndrome,GH and POMC were down-regulated in asthenia of qi syndrome,ATG and Gal were up-regulated between asthenia of yang and qi syndrome and asthenia of qi and yin and yang syndrome.However,FSH was down-regulated coinstantaneously in different asthenia syndromes,and PACAP,CCK and NT were down-regulated remarkably.[Conclusion]Neuropeptide genes expression are selective in adrenal gland of mice.A part of neuropeptides are different expression model in different syndromes and signs of H22 tumour mice,such as GH,POMC and Gal gene expression could be related to syndromes and signs,however,PACAP,CCK and NT could be related to H22 tumour disease.

目的:观察荷瘤小鼠肿瘤发生后不同阶段4个常见证候睾丸独特上、下调的基因.方法:采用荷瘤小鼠及其标准化四诊与辨证方法,及GeneChip Mouse Exon ST 1.0 Array等技术,检测早期邪毒壅盛证及气虚证、中期阳气虚证、中晚期气阴阳虚证睾丸基因的差异表达,重点关注那些表达较高的基因.结果:共捡出独特上调和下调的基因28个,其中有5个基因在不同证候中上下调不一致.肿瘤发生后早期邪毒壅盛证上调的基因8个:Abpg、Pip、Muc10、Car6、2310057J18Rik、Klk1b3、Klk1、Wfdc12,下调的基因5个:Spcs1、Dtx3、Rasa3、Sec11l3、F5;早期气虚证上调的基因4个:Gpx5、Wbp5、Lcn5、Ap1s2,下调的基因1个:Klk1b26;中期阳气虚证无上调基因,下调基因3个,分别为Klk1b27、Spag5、Gpx5(在早期气虚证上调);中晚期气阴阳虚证上调基因2个,分别为Saa3、S100a8,下调基因5个;Abpg、Pip、Muc10、Car6、Cuzd1(前4个在邪毒壅盛证上调).结论:荷瘤小鼠在早期阶段,邪毒壅盛证与睾丸基因表达的紊乱关系较为密切;中期阳气虚证、中晚期气阴阳虚证睾丸基因以下调为主,提示中、晚期虚实夹杂证的荷瘤小鼠其睾丸功能下降.

Objective To observe the general characteristics of gene transcription of neuroendocrine and immune tissues of common syndromes in tumor-bearing mice. Methods Standardized diagnostic and syndrome differentiation methods for mice,as well as GeneChip Mouse Exon 1.0 ST Array were employed to detect the RNA transcription and splicing in the hypothalamus,hypophysis,adrenal gland,testicle,spleen and thymus of four syndromes in H22-bearing mice.Results The 28-second peaks of RNA electrophoresis of hypothalamas,hypophysis and adrenal gland were higher than 18-second peaks in Kunming male mice;vice versa in the testicle,spleen and thymus.In the early stage of tumor,gene expressions in hypothalamus,hypophysis and adrenal gland changed dramatically,especially in toxin syndrome;28 s RNA in hypothalamus elevates rapidly and continues,more obvious in qi-deficiency syndrome than in toxin syndrome;total amount of RNA in hypophysis decreased more abruptly in 28 s,especially in toxin syndrome.RNA electrophoresis of adrenal gland was similar to that of hypophysis,with similar electrophoresis in qi-deficiency syndrome and in toxin syndrome,but differed in qi-yin-yang-deficiency syndrome in late tumors.No striking changes were found in testicle.With the progression of tumor,the weight of the spleen increased continuously,while the weight of thymus decreased continuously;the protein synthesis and sugar metabolism reduced,with more protein synthesis and sugar metabolism in toxin syndrome than in qi-deficiency syndrome.Compared with the control group,9 127 gene expressions and 51 126 exons splicing in above seven types of tissues experienced striking differences.Conclusion The difference of gene transcription of neuroendocrine and immune tissues is the vital material foundation of syndrome of tumor-bearing mice.

目的:筛选荷瘤小鼠早期垂体差异表达的基因,并简要分析其功能特征。方法:采用荷瘤小鼠标准化四诊及辨证方法,及GeneChip Mouse Exon1.0STArray等技术,检测H22荷瘤小鼠早期邪毒壅盛证和气虚证、中期阳气虚证、中晚期气阴阳虚证共3阶段4个证候垂体基因表达的差异,重点关注早期垂体差异表达的基因。结果:筛选到早期上调的基因12个:Alb1、Timp3、Ifi44、Kdr、Cbln3、Hectd2、Hecw2、Vtn、LOC631028、LOC630776、4933427I04Rik、A630075K04Rik。下调的基因17个:Klk1b27、Klk1b26、Klk1b4、Klk1b22、Klk1b3、Klk1、Klk1b16、Muc10、Cryaa、Bag3、Gsg2、Six2、Saa3、1700108L22Rik、LOC628820、2310057J18Rik、2410002I01Rik。结论:H22荷瘤小鼠早期垂体基因表达改变集中表现为保护蛋白的功能减弱、细胞周期改变,以及血管、神经、表皮生成存在障碍的倾向,这可能与荷瘤小鼠早期垂体应激代偿,甚至是失代偿有关。其中Cbln3可能是邪毒壅盛证的标志基因之一。

目的:探讨H22荷瘤小鼠肿瘤发生后睾丸基因表达谱改变的特征.方法:采用荷瘤小鼠标准化四诊及辨证方法.以及Gene Chip Mouse Exonl.0 ST Array等技术,检测H22荷瘤小鼠睾丸组织早期邪毒壅盛证和气虚证、中期阳气虚证、中晚期气阴阳虚证3个阶段4个证候睾丸基因表达谱的差异,重点关注肿瘤发生后一致上调或下调明显且表达量大的基因.结果:肿瘤发生后三个阶段一致上调的基因5个;一致下调的基因12个,其中7个与性激素和生殖有关;早期上调、中期和中晚期下调的基因1个;早期和中期上调、中晚期下调的基因3个.在这些基因中,Saa3在晚期气阴阳虚证表达上调了近60倍,Abpg、Pip、Car6在早期邪毒壅盛证表达上调了4|D一80倍,值得关注.结论:肿瘤发生后荷瘤小鼠睾丸基因表达的改变以下调为主,性激素合成与生殖受到影响;一些表达显著改变的基因值得进一步研究.

我们在以往的研究中发现,某些疾病模型小鼠,如肿瘤小鼠,存在类似人类的证候及演变[1～4].

目的:观察辨证论治对H22荷瘤小鼠的生存质量、生存期的影响。方法:将雄性昆明种小鼠随机分组,除正常对照组外,腋下接种H22瘤株,出瘤后,分层随机分为模型组、非辨证治疗组(以下简称非辨组)、辨证治疗组(以下简称辨证组)。采用小鼠四诊标准化、计量化采集方法,收集相关诊断指标,进行辨证,非辨组给予固定方治疗,辨证组根据具体证型给予相应的辨证论治。综合评估其生存质量及生存期限。结果:用药组生存期均长于模型组;辨证组各项四诊指标和证候的严重程度优于非辨组、模型组。结论:荷瘤小鼠是可以实现个体化辨证论治的,经辨证论治对其生存质量及生存期均有帮助。

目的:筛选荷瘤小鼠阳气虚证垂体差异表达的基因,并简要分析其功能特征.方法:采用荷瘤小鼠及其标准化四诊及辨证方法,及GeneChip Mouse Exon1.0 ST Array等技术,检测H122荷瘤小鼠早期邪毒壅盛证和气虚证、中期阳气虚证、中晚期气阴阳虚证共3阶段4个证候垂体基因表达的差异,重点关注那些表达量大、差异显著的基因,筛选阳气虚证差异表达的基因.结果:筛选到阳气虚证独特上调的基因14个:LOC669166、Cd8b1、Cd3d、Ly6d、Lat、Orm2、Dntt、1700021K02Rik、A130072A22Rik、IDC669782、Ptpre、Igk-V21-9、Cxcll3、LDC668417,其中多数基因在正常小鼠及其他证候小鼠的垂体表达很低或几乎关闭;独特下调的基因3个:Wfdc1、Spook3、Mfan4.结论:H22荷瘤小鼠阳气虚证垂体基因表达与其他证候存在较大差异,其中部分可能是阳气虚证的标志基因.

我们以往的研究一再发现,荷瘤小鼠在肿瘤发生后,会自发形成证候及其演变,早期以邪毒壅盛证、气虚证多见,且发现邪毒壅盛证动物预后差,带瘤生存时间短.鉴于学术界以往的研究观察到中医证候的形成与甲状腺密切相关,那么,邪毒壅盛证发生时,甲状腺是否存在独特的基因表达特征,是否与该证候形成有关?因此,我们通过业已建立的小鼠四诊工作站及其系列标准[1-2],对H22荷瘤小鼠进行辨证,筛选出早期邪毒壅盛证、气虚证、中期阳气虚证、中晚期气阴阳虚证等4个常见证型,继而采用GeneChip Mouse Exon 1.0 ST Array等技术对以上各证候荷瘤小鼠甲状腺等组织的基因进行了检测,本文重点报道邪毒壅盛证甲状腺独特上调与下调且表达量较高的基因.

Objective:To reveal the character of highly expressed Genes in different stage from the thyroid of H22 Tumour Mice.Meth-ods:By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array,we observed thyroid gene expression about poisonous pathogenic factors syndrome and asthenia of qi syndrome on earlier period tumour mice.Highly expressed highly are analyzed in the paper.Results:in early period,there are 1 up-regulated gene lfi27 and 2 down-regulated high expressed genes,namely AY036118 and Dub2a in middle stage,25 up-regulated high expressed genes were Ktklb27,Crispl,Fas,H2-Ea and GOs2.In in-termediate and advanced stage,there are 1 up-regulated and 3 down-regulated high expressed genes,the former namely Miff,the latter kind namely Sparc,Co14al and Co13al.Conclusion The expression and functional characteristics of some genes present certain ternds,for exam-ple,in middle stage,the up-regulated high expressed genes are much more than that in other two stages.These genes involved in cellular ar-chitecture,movement,proliferation,apoptosis may be related to Tumorigenesis.In these genes above,most genes were less researched in thyroid physiology and pathology.They function in thyroid and the thyroid function in Tumorigenesis expecting more study.

Objective: To investigate the characters of gene expression in adrenal gland of H22 tumor mice of yang-qi deficiency syndrome and qi-yin-yang deficiency syndrome. Methods: The adrenal gland gene expression in H22 tumor mice of yang-qi deficiency syndrome on intermediate stage,qi-yin-yang deficiency syndrome on advanced stage were detected with quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array. Results:①52 up-regulated genes and 11 down-regulated genes were obtained on yang-qi deficiency syndrome,23 up-regulated genes and 4 down-regulated genes were obtained on qi-yin-yang deficiency syndrome.②Up-regulated genes on yang-qi deficiency syndrome including proteolysis genes,such as Klk1,Ctrc,Ctrb1,Ren1,Ela3,Cpa1,Prss2,Cpb1; lipid metabolism genes,such as Clps,Pnlip,Pnliprp1,Cel; spermatogenesis genes,such as Tnp2,Tnp1,Crisp1,Prm1,Prm2,Smcp; hormone correlated proteinum genes,such as Pip,Abpg. Down-regulated genes on yang-qi deficiency syndrome involve in lipid metabolism genes,such as Scd1,Fasn,Acaca,Acss2,Pnpla3.③Up-regulated genes on qi-yin-yang deficiency syndrome including inflammatory response genes,such as S100a8,S100a9,Alox5ap,Orm1,Kng1,Mpo; hemagglutinin and cell proliferation regulated genes,such as Fga,Fgb,Fgg,Prtn3,Fgl1; ion transport genes,such as Lcn2,Hpxn,Ltf,and so on. Down-regulated genes on qi-yin-yang deficiency syndrome only Abpg,Klk1b22,Klk1 and Reg1. Conclusion: Gene expressions change a lot in adrenal gland of H22 tumor mice of yang-qi deficiency syndrome and qi-yin-yang deficiency syndrome,which form the intrinsic features of the two syndromes.

OBJECTIVE:To reveal the characteristics of gene expression in adrenal gland of H22 tumor mice with typical syndromes and in different liver cancer stages.METHODS:By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expression in H22 tumor mice with pathogenic factor-toxin predominance syndrome and qi deficiency syndrome in the earlier stage, yang-qi deficiency syndrome in the intermediate stage, and qi-yin-yang deficiency syndrome in the advanced stage. Genes highly expressed and remarkably different were analyzed in this study.RESULTS:A total of seventy-three up-regulated coincident genes and twenty-six down-regulated coincident genes in different stages were investigated in the study. Up-regulated coincident genes included Hp, C3, Anxa1, Procr, C2, Il4ra, Cd14, Ptprc, Cd52, C4b, Eno3, Xdh, Gpx3, and so on. Down-regulated coincident genes included nervous system function-related genes such as Plp1, Mbp, Aldh1a1, Cck, Atn1, genes associated with electrolyte metabolism such as Aldh1a1 and Slc22a17, genes related to signal transduction such as Cxcr4, Spag5 and Stmn3, etc, and genes related to transcriptional control and protein biosynthesis such as Hspa1a, Dnajb1, Thra, Hhex and so on.CONCLUSION:With the development of the tumorigenesis, the symptoms and signs and differentially expressed genes in adrenal gland of H22 tumor mice can be measured. Up-regulated and down-regulated coincident genes may be the features of H22 tumor mice different from those of normal mice.

Objective:To observe the pathological change of the common used gastric ucer mice model,to appraise and compare these various models.Methods:We select six methods:stress,acetic acid,histamine phosphate,antifani,apoplon,alcohol to make gastric ulcer model simultaneously,and set up the normal,abrosia,and sham operated control groups.We get the stomach to observe pathohistology on the 1st and 8th day.Results:On the 1st day,there were conspicuous pathological changes in the abrosia,stress and sham operated groups mice stomach which disappeared on the eighth day.In the antifani,apoplon and alcohol groups obvious pathological changes were found on the 1st day and lasted till the 8th day.The histamine phosphate group's pathological changes were slight and easy to recover.The acetic acid seriously damaged the animal and easily to resulted in death.The Above mentioned pathological changes,which were mostly mucous membrane partial or diffuse inflammation,dropsy,hemorrhage,or the mucous membrane atrophia,the necrosis and amotic,were the acute gastritis or the partial acute gastric ulcer.Conclusion:The stress,antifani,apoplon,and alcohol model highly successful ratio.The abrosia and sham operated control groups were the next,but the histamine phosphate and the acetic acid group seem not to suit the mouse conventional gashtric ulcer model.

Objective To reveal the character of high expression genes in adrenal gland of normal and H22 tumour mice about typical symptoms and signs. Methods By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expression about poisonous pathogenic factors syndrome and asthenia of qi syndrome on 8th day, asthenia of yang and qi syndrome on 21th day, asthenia of qi and yin and yang syndrome on 29th day with H22 tumour mice. Results 402 high expression genes were gained, and the computed value of 88genes are more than 5 000. These genes participate in metabolism, gene expression and regulation, chemistry synthesis and processing and modify, transport, signal transduction, immunization, cell cycle, apoptosis, development, and some genes function are unknown. Conclusion Those genes are persistent and steady expression in normal and H22 tumour mice adrenal that could be important biology significance. Genes expressed down-regulate may be reflect the essence and feature of asthenia of qi syndrome. Otherwise, differentially expressed genes in adrenal gland have coherent tendency among four different symptoms and signs, which confirm the theory and essence about different symptoms and signs of the same disease, and it is essential to execute syndrome differentiation and treatment.

Objective: To investigate the characteristics of gene expression in pituitary of H22 tumor-bearing mice of different syndromes. Methods: The pituitary gene expression in H22 tumor-bearing mice of pathogenic toxin-exuberance syndrome and qi-deficiency syndrome on early stage, yang-qi deficiency syndrome on intermediate stage, qi-yin-yang deficiency syndrome on advanced stage were detected with quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array to screen out the up- and down-regulated genes in uniform. Results: There are twenty-three up-regulated genes including Alas2, Ube2l6, Sgk,BC055107, Cdh26, Bhlhb2, Nr1d2, Slc39a14, Serpina3n, Ms4a6c, Lcp1, Dhx8, Il1r1, S100a8, S100a9, Cxcl1, Hp, Csf1r, RIKEN cDNA 2610307008 gene, Phactr3, Mknk1, Aloxe3 and Aqp11, and nine down-regulated genes including 2310047A01Rik, Hoxa10, Cryaa, Hspa1a, Dub2a, Mpz, Klk1, Naaladl1, Prph1, some of which have significant changes in certain syndromes. Conclusion: There are significant differences in gene expression of pituitary of H22 tumour-bearing mice, some of which may relate to syndrome types.