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RESULTS A eukaryotic expression system for high expression humanmutantCD59 were successfully set up : The recombinant PALTER-MAX plasmid containing human mutantCD59 cDNA and PCDNA plasmid were co-transfected into CHO cell by cation lipoid mediating method ;and the cells were grown in F12 medium containing 400ug/ml G418 for 14 days, positive clones were grown in RPMI1640 medium to get stable expressing cell lines . Highly expressing clones were selected by flow cytometry ,and were named PALTER-CD59-CHO1PALTER-CD59-CHO2 . Flow cytometry indicated that expression rates of PALTER-CD59-CHO1 and PALTER-CD59-CHO2 were 53.7%and 54.5%. Further more, Stable highly expressing CHO cell lines were more detected by immunocytochemistry and immunofluorescence technology . PALTER-CD59 -CHO1 and PALTER-CD59-CHO2 were grown in RPMI1640 to get a large of cells . CD59 protein were obtained by spalling PALTER- CD59- CHO1 and PALTER - CD59 - CHO2 cells . Stable highly expressing cells were further validated by SDS-PAGE, immunoblot analysis and solid enzyme immunoassay . PALTER - CD59 - CHO1 and PALTER - CD59 - CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER-CD59-CHO1 or PALTER-CD59-CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59 -CHOI or PALTER-CD59-CHO2 was higher than unglycated ones . PALTER -CD59-CHO1 and PALTER -CD59 -CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER - CD59 - CHOI or PALTER - CD59 - CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59-CHO1 or PALTER - CD5 9-CHO2 was higher than unglycated ones .

结果 成功构建突变人CD59的真核细胞表达系统：运用阳离子脂质体介导法将含有突变人CD59的PALTER—MAX重组质粒与PCDNA共转染入CHO细胞：用含有400ug／mlG418的F12培养基培养14天，筛选出稳定阳性表达克隆，RPMI1640培养基扩增获得稳定表达细胞株，并用流式细胞术进一步筛选出高效表达细胞株分别命名为PALTER—CD59—CH01、PALTER—CD59—CH02，表达率分别为53.7％、54.5％；应用免疫组化方法、免疫荧光技术进一步鉴定阳性细胞株；RPMI1640培养基大量扩增PALTER—CD59—CH01、PALTER—CD59—CH02细胞株，裂解细胞得到CD59蛋白质；通过SDS—PAGE凝胶电泳技术、免疫印迹技术、固相酶联免疫吸附试验验证了这两中文摘要个阳性细胞株CO59蛋白的高效表达；50mM核糖培养72小时，获得突变人CD59糖化细胞株，BCECF染料释放试验结果显示，PALTER一CD59一CHOI、pALTER一CD59一CHOZ细胞较PALTER一CHO细胞染料释放率低，未糖化PALTER一CD59一CHOI、PALTER一CD59一CHOZ细胞比较糖化后细胞染料释放率低。

Major results were summarized as follows:(1) A total of 58 tree species from 53 genera in 36 families among which 8 species were Rosaceae, 5 species were Liliaceae, 5 species were Eriaceae, 3 species were Ranunculaceae, 3 species were Compositae, 2 species were Cupressaceae, 2 species were Caprifoliaceae, 2 species were Umblliferae, and other 28 tree species were only one receptively. These were recorded in the 8 plots of the A. georgei var. smithii forest;(2) Number of families, genera, species and Margalef index correlated negatively with altitude (P.05), with a peak at 3600 m.(3) Shannon-Wiener index correlated negatively with altitude (P.01), and maintained stable at the altitudes between 3700~4100 m, evenness with altitudes, however, this trend was insignificant.(4) Jaccard index increased sharply with increasing altitude at the altitudes between 3600~4100 m, and was lower between different vegetation types at the altitudes between 4100~4200. Cody index β(subscript c decreased with an increasing altitude, but there were 2 troughs between 4000~4100 m and 4200~4300 m.(5) Maximum tree height H(subscript max and HH(subscript max=37 mcorrelated negatively with altitude (P.05); but basal area and BA BA(subscript max=5.3m^2 correlated with altitudes, however, this trend was insignificant.

结果表明：(1)在调查的8个急尖长苞冷杉林样地内共有植物58种，分属于36科53属，其中植物较多的科有蔷薇科8种、百合科5种、杜鹃花科5种、毛茛科3种、菊科3种、柏科2种、忍冬科2种、伞形科2种，其余的28科各只有1个种；(2)物种科、属、种数、Margalef指数D(下标 M与海拔存在显著的负相关性(P.05)，在分布急尖长苞冷杉最低海拔3600m处出现物种丰富度的最大值；(3)多样性指数与海拔之间有极显著的负相关性(P.01)，并且在3700~4100m之间多样性指数保持稳定；均匀度指数与海拔梯度之间存在负相关性，但不显著；(4)Jaccard指数C(下标 j在海拔3600~4100m随海拔的升高而升高，在生境过渡带的4100~4200m之间Jaccard指数C(下标 j较低；Cody指数β(下标 c随海拔的升高呈下降的趋势，但在4000~4100m和4200~4300m 海拔区间出现2个低谷；(5)最大树高H(下标 max和最大胸径DBH(下标 max与海拔之间存在显著负相关性(P.05)；胸高断面积之和和立木密度与海拔之间存在负相关性，但不显著。

Compared with control,①the mean tumor weight of H22 of SXKA Granules three dose groups were decreased significantly(P.01,P.05),and the mean inhibition rates of SXKA Granules 20、10 g/kg dose groups were above 30%;②the mean tumor weight of S180 of SXKA Granules three dose groups were decreased significantly(P.01),and the mean inhibition rates of SXKA Granules 20、10 g/kg dose groups were above 32%;③the mean tumor weight of EAC of SXKA Granules three dose groups were decreased significantly ( P.01, P.05),and the mean inhibition rates of SXKA Granules 20 g/kg dose groups were above 38%;④the mean tumor weight of Lewis carcinoma of SXKA Granules three dose groups were decreased significantly(P.01, P.05),and the mean inhibition rates of SXKA Granules 20、10、5 g/kg dose groups were above 36%;⑤the mean tumor weight of W256 of SXKA Granules three dose groups were decreased significantly ( P.01, P.05),and the mean inhibition rates of SXKA Granules 20、10 g/kg dose groups were above 32%;⑵Compared with control,SXKA Granules 20、10 g/kg dose groups had extended the survial time of the P388-bearing mice respectively(P.01),and the mean prolong rates of SXKA Granules 20、10 g/kg dose groups were above 50%;⑶Compared with S180-bearing group, SXKA Granules 20、10 g/kg dose groups could increase the weight of thymus and spleen, Spleen index and thymus index were increased, SXKA Granules 5 g/kg dose group could increase thymus index(P.05);⑷As Compared with control group, SXKA Granules 20、10 g/kg dose groups could improve mouse serum half hemolysis value depressed by transplanted tumor dramatically(P.01), which revealed the SXKA granules could improve the mouse humoral immunity system;⑸SXKA Granules 20 g/kg dose group could increase of englobe indexαon S180-bearing mice remarkably(P.01), which indicated the SXKA Granules could improve their cellular immunity system.

对荷W256大鼠，生兴克癌冲剂20、10、5 g / kg三组的平均瘤重明显低于对照组(P.01，P.05)，生兴克癌冲剂20、10 g / kg组的平均肿瘤抑制率均大于32 %；⑵与空白对照组相比，生兴克癌冲剂20、10 g/ kg能显著地延长移植小鼠白血病P388小鼠的存活天数(P.01)，生兴克癌冲剂20、10 g/ kg对荷白血病P388小鼠生命延长率均在50%以上；⑶与S180荷瘤组相比，生兴克癌冲剂对荷瘤鼠的免疫器官重量、胸腺指数和脾指数有一定的提高趋势，其中生兴克癌冲剂5 g / kg组对荷瘤小鼠的胸腺指数有一定的提高作用(P.05)；⑷与S180荷瘤组相比，生兴克癌冲剂20、10 g /kg组可提高由荷瘤引起的小鼠血清半数溶血素值的降低(P.01)，表明其可提高荷瘤小鼠体液免疫功能；⑸与对照组相比，生兴克癌冲剂20 g /kg组可提高荷S180肉瘤小鼠的免疫吞噬系数α值(P.01)，表明其可提高荷瘤小鼠细胞免疫功能。

Thecompare of genetic map between Lowes and ours showed 26 homology marker situ,which occupied 21.1% of the marker situ in the experiment. 81 QTLs were detected for 11 agronomic traits. 4 QTLs were detected for plantheight, which explained 10.3%～28.9% of trait variance; 2 QTLs were detected forNo. of effective 1-st branches, which explained 22.1%～47% of trait variance; 16QTLs were detected for effective branches height, which explained 12.2%～51.8% oftrait variance; 15 QTLs were detected for length of main inflorenscence, whichexplained 7.4%～26.6% of trait variance; 5 QTLs were detected for effective siliquesof main inflorenscence, which explained 11.2%～25% of trait variance; 1 QTLs weredetected for density of main infiorenscence, which explained 17.3% of trait variance;12 QTLs were detected for length of silique, which explained 24%～36.7% of traitvariance; 2 QTLs were detected for seed per sillique, which explained 9.6% and16.9% of trait variance; 2 QTLs were detected for 1000 seed weight, which explained26%～13.7% of trait variance; 11 QTLs were detected for Total effective siliques perplant, which explained 14.8%～47.2% of trait variance; 11 QTLs were detected forplant height, which explained 14.3%～32.8% of trait variance.

其中，株高检测到4个QTLs，解释性状表型变异的10.3％～28.9％；一次有效分枝数检测到2个QTLs，解释性状表型变异的22.1％和47％；有效分枝部位检测到16个QTLs，解释性状表型变异的12.2％～51.8％；主花序长度检测到15个QTLs，解释性状表型变异的7.4％～26.6％；主花序有效角数检测到5个QTLs，解释性状表型变异的11.2％～25％；主花序角密度检测到1个QTLs，解释性状表型变异的17.3％；角果长度检测到12个QTLs，解释性状表型变异的24％～36.7％；每角粒数检测到2个QTLs，解释性状表型变异的9.6％和16.9％；千粒重检测到2个QTLs，解释性状表型变异的26％和13.7％；单株有效角果总数检测到11个QTLs，解释性状表型变异的14.8％～47.2％；单株产量检测到11个QTLs，解释性状表型变异的14.3％～32.8％。

Results: Amang 128 patients, 56 cases were diagnosed as pulmonary carcinoma (43.75%), of which 30 cases were squamous carcinoma (53.57%), 22 cases were adenocarcinoma (39.29%), 4 cases were small cell undifferentiated carcinoma (7.14%); 37 cases were bronchial pneumonia (28.91%), in which 30 cases were given brush sampling, bacterial culture of BALF, the detectable rate of pathogenic bacteria were 50%(15 cases) and 40%(12eases) respectively; 12 eases were pulmonary tuberculosis (9.38%), of which 3 eases (25%) had concurrent endobronchial tuberculosis, smear of brush biopsy and BALF found Mycobacterium tuberculosis were 11 eases (91.67%) and 5 eases (41.67%) respectively, 2 eases were diagnosed as caseous tubereulous granuloma (16.67%); 2 eases were idiopathic pulmonary fibrosis (1.56%), lease was lymphoma of long.

结果：128例患者确诊为肺癌56例(43.75%)，其中鳞癌30例(53.57%)，腺癌22例(39.29%)，小细胞癌4例(7.14%)；支气管肺部炎症病变37例(28.91%)，其中30例经防污毛刷采样、支气管肺泡灌洗液行细菌学培养，病原菌的检出率分别为50%（15例）和40%（12例）；肺结核12例(9.38%)，其中合并支气管内膜结核3例(25%)，刷检涂片和支气管肺泡灌洗液找到结核杆菌分别为11例(91.67%)和5例(41.67%)，病理确诊为结核性乾酪样肉芽肿2例(16.67%)；特发性肺间质纤维化2例(1.56%)，肺淋巴瘤1例(0.78%)。

Twelve tumors were solid and three contained cystic components. Nine were inhomogeneously hypoechoic. Two were homogeneously hypoechoic. Two were inhomogeneously isoechoic. Two were homogeneously isoechoic. Two contained microcalcification. One with acoustic shadowing. Twelve were well-defined and three were ill-defined. Fourteen were regular and one was irregular. Four had acoustic halo. Six were ecstatic. Nine were not ecstatic. Thirteen were examined with color Doppler ultrasound. Nine had abundent branched blood flow within the tumor and ring blood flow around. Two had short linear blood flow within the tumor and ring flow around.

不均质低回声9个，均匀低回声2个，不均质中等回声2个，均匀中等回声2个。2个肿瘤内见细小钙化斑，1个伴声衰减。12个肿瘤边界清，3个边界欠清。14个肿瘤形态规则，1个形态不规则。4个肿瘤伴&晕环征&。6个肿瘤有膨胀感，9个肿瘤无明显膨胀感。13个肿瘤行彩色超声检查，9个内部见丰富分枝状血流，周边见环状血流，2个内部见短线状血流，周边环状，仅2个内部无彩色血流，周边见点状彩色血流。

Objective To analyze the CT and MRI manifestations of oral carcinoma,and to evaluate the value of imaging technologies in the diagnosis of oral carcinoma,thus to explore the most reasonable imaging method Methods 46 cases with oral carcinoma,proved by surgery and pathology （2 lip tumors,8 carcinomas of the gingival ridge,5 carcinomas of the buccal mucosa,24 tongue carcinomas,5 carcinomas of the floor of the mouth,2 tumors of the hard palate）,were analysed retrospectively32 cases were examined with CT and 14 cases with MRIImaging presentations were compared with surgery and pathology Results Of the 46 cases with oral carcinoma,22 were ulcerative lesion of infiltrative growth,24 were masslike lesion of extruded trowthOf the 32 cases examined with CT scan,27 were detected,5 were missed,the 4 missed cases were confirmed to be superficial ulcers by surgery and pathologyOf the 10 cases with bone destruction examined with CT scan,8 were manifested,2 were missedAnd of the 14 cases with MRI scan,all 14 were detected Conclusion CT and MRI scan are valuable for diagnosis of oral carcinomasThey can manifest the morphology and size of primary tumor,contiguous structures,bone invasion and cervical lymph node metastasisMRI is the optimal imaging method for superficial ulcerative lesion of infiltratibe growth

徐 锋，罗德红，唐威，欧阳汉，周纯武目的分析口腔癌的CT及MRI表现，评价影像检查在口腔癌诊断中的应用价值，探索合理的影像学检查方法。方法回顾性分析经手术及病理证实的口腔癌46例（唇癌2例，牙龈癌8例，颊黏膜癌5例，舌癌24例，口底癌5例，硬腭癌2例），其中32例行CT扫描，14例行MRI扫描，将影像表现与手术所见及病理结果进行对照分析。结果 46例口腔癌中，呈浸润性生长的溃疡型病变22例，外生性生长的肿物型病变24例。CT扫描32例，明确显示病变27例，遗漏病变5例，其中4例为表浅溃疡型病变。CT扫描病例中，10例有骨质侵犯，CT扫描准确评价8例，遗漏2例。MRI扫描14例，均准确评价肿瘤及其与周围结构的关系。结论 CT、MRI对口腔癌原发肿瘤的形态、大小、周围结构侵犯、颈部淋巴结转移的评价有重要价值。对于较小及呈浸润性生长的表浅溃疡型肿瘤，CT扫描尤其是CT平扫难以准确显示，应首选MRI。

Objective To analyze the CT and MRI manifestations of oral carcinoma,and to evaluate the value of imaging technologies in the diagnosis of oral carcinoma,thus to explore the most reasonable imaging method Methods 46 cases with oral carcinoma,proved by surgery and pathology （2 lip tumors,8 carcinomas of the gingival ridge,5 carcinomas of the buccal mucosa,24 tongue carcinomas,5 carcinomas of the floor of the mouth,2 tumors of the hard palate）,were analysed retrospectively32 cases were examined with CT and 14 cases with MRIImaging presentations were compared with surgery and pathology Results Of the 46 cases with oral carcinoma,22 were ulcerative lesion of infiltrative growth,24 were masslike lesion of extruded trowthOf the 32 cases examined with CT scan,27 were detected,5 were missed,the 4 missed cases were confirmed to be superficial ulcers by surgery and pathologyOf the 10 cases with bone destruction examined with CT scan,8 were manifested,2 were missedAnd of the 14 cases with MRI scan,all 14 were detected Conclusion CT and MRI scan are valuable for diagnosis of oral carcinomasThey can manifest the morphology and size of primary tumor,contiguous structures,bone invasion and cervical lymph node metastasisMRI is the optimal imaging method for superficial ulcerative lesion of infiltratibe growth

中华硕博网核心提示：徐锋，罗德红，唐威，欧阳汉，周纯武关键词：口腔肿瘤目的分析口腔癌的CT及MRI表现，评价影像检查在口腔癌诊断中的应用价值，探徐锋，罗德红，唐威，欧阳汉，周纯武目的分析口腔癌的CT及MRI表现，评价影像检查在口腔癌诊断中的应用价值，探索合理的影像学检查方法。方法回顾性分析经手术及病理证实的口腔癌46例（唇癌2例，牙龈癌8例，颊黏膜癌5例，舌癌24例，口底癌5例，硬腭癌2例），其中32例行CT扫描，14例行MRI扫描，将影像表现与手术所见及病理结果进行对照分析。结果46例口腔癌中，呈浸润性生长的溃疡型病变22例，外生性生长的肿物型病变24例。CT扫描32例，明确显示病变27例，遗漏病变5例，其中4例为表浅溃疡型病变。CT扫描病例中，10例有骨质侵犯，CT扫描准确评价8例，遗漏2例。MRI扫描14例，均准确评价肿瘤及其与四周结构的关系。结论 CT、MRI对口腔癌原发肿瘤的形态、大小、四周结构侵犯、颈部淋巴结转移的评价有重要价值。对于较小及呈浸润性生长的表浅溃疡型肿瘤，CT扫描尤其是CT平扫难以准确显示，应首选MRI。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

But this is impossible, as long as it is engaging in a market economy, there are risks in any operation.

但是，这是不可能的，只要是搞市场经济，是有风险的任何行动。

We're on the same wavelength.

我们是同道中人。