反义N-ras-1基因对成纤维细胞生长因子诱导的血管平滑肌细胞增殖的抑制作用

目的应用反义Ｎ－ｒａｓ１基因阻断成纤维细胞生长因子（ｂＦＧＦ）的促增殖作用，为防治以血管平滑肌细胞（ＶＳＭＣ）增殖为主要特征的血管损伤后再狭窄等心血管疾病提供进一步探索的途径。方法利用构建的含反义Ｎ－ｒａｓ１基因的逆转录病毒质粒（ｆｐＧｖ１－ＭＴ－ＡｎｔｉｓｅｎｓｅＮ－ｒａｓ１）转染于培养的血管平滑肌细胞（ＶＳＭＣｓ），ｆｐＧｖ１－ＭＴ逆转录病毒空载质粒及未转染质粒的细胞为对照，观察其对ｂＦＧＦ诱导的ＶＳＭＣ的影响。结果反义Ｎ－ｒａｓ１细胞数为（１６．８±１．３）×１０４（细胞／ｍｌ），空载质粒对照细胞为（３０．１±１．２）×１０４，两者差异有显著意义（Ｐ＜０．００１）；反义Ｎ－ｒａｓ１细胞３Ｈ－ＴｄＲ掺入率为７６４３±６７２（ｃｐｍ），空载质粒对照细胞为１５１３１±１３８（ｃｐｍ），两者差异有非常显著意义（Ｐ＜０．００１），同时应用反义Ｎ－ｒａｓ１基因的细胞ＲａｓｍＲＮＡ表达水平明显降低，其表达蛋白ｐ２１也明显降低。结论反义Ｎ－ｒａｓ１基因对ＶＳＭＣ及ｂＦＧＦ诱导ＶＳＭＣ增殖有抑制作用。     

分泌型磷脂酶A2 cDNA与载体pRc／CMV定向克隆及表达

为了构建分泌型磷脂酶Ａ２（ｓｅｃｒｅｔａｒｙ ｐｈｏｓｐｈｏｌｉｐａｓｅＡ２,ｓＰＬＡ２）ｃＤＮＡ的有效表达系统,该文从无效表达的重组体ｓＰＬ０Ａ２－ｐＧＥＭ７中制备ｓＰＬＡ２片段,将其插入中间载体Ｂｌｕｅｓｃｒｉｐｔ ＢＳＳＫＲ的ＥｃｏＲ１位点,利用ｓＰＬＡ２－ＢＳＳＫ重组体转化ＤＨ１感染态细胞,筛选ａｎｔｉｓｅｎｓｅ ｓＰＬＡ２－ＢＳＳＫ重组体,应用Ｘｂａｌ、Ｈｉｎｇｄ３双酶消化该重组体并回收     

TGFβ1反义寡核苷酸对大鼠肝脏星状细胞激活及其胶原生成的 …

为探讨ＴＧＦβ１反义寡核苷酸（ａｎｔｉｓｅｎｓｅ ｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓ,ＡＳＯＮ）对大鼠肝脏星状细胞活化及其胶原生成的作用,作者合成了特异笥的ＴＧＦβ１硫代磷酸ＡＳＯＮ及其对照（正义,错义寡核苷酸）,并将其加入至培养的肝脏星状细胞中。通过细胞免疫化学方法分析α－ＳＭＡ的表达来观察肝脏星状细胞的活化,肝脏星状细胞产生ＴＧＦβ的水平用生物学方法测定,以＾３Ｈ－脯氨酸掺入抑制率的测定观察胶原     

阳离子脂质体介导的TGFβ1反义寡核苷酸对大鼠肝脏星 …

为探讨阳离子脂质体介导的ＴＧＦβ１反义寡核苷酸（ａｎｔｉｓｅｎｓｅ ｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓ，ＡＳＯＮ）对大鼠肝脏星状细胞激活及其胶原生成的作用，作者合成了特异性的针对ＴＧＦβｍＲＮＡ转译超始区的硫代磷酸ＡＳＯＮ及其对照（正义、错义寡核苷酸），并和阳离子脂质体形成复合物。通过测定细胞内＾３２Ｐ标记的ＡＳＯＮ的放射量来观察ＡＳＯＮ的细胞摄入率，用细胞免疫化学分析α－ＳＭＡ的表达来观察星状细胞     

弓形虫主要表面抗原基因克隆及在大肠杆菌中的表达

为在大肠杆菌中表达弓形虫主要表面抗原（ＳＡＧ１）,进一步研究其生物学功能。将ＳＡＧ１基因重组于ｐＧＥＭＥＸ－１融合型表达载体上,转化大肠杆菌ＪＭ１０９（ＤＥ３）,得到含重组表达质粒ｐＧＥＭＥＸ－１／ＳＡＧ１的工程菌。结果表明ＩＰＴＧ诱导表达后,经ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ－ｂｌｏｔ检测证实含有ＳＡＧ１蛋白。提示ｐＧＥＭＥＸ－１大肠杆菌系统可以表达具有免疫活性的ＳＡＧ１蛋白。     

Effect of Buyang Huanwu Decoction (补阳还五汤) on Plasma Thromboxane B_2, 6-Keto-Prostaglandin F_(1α), Endothelin and Calcitonin Gene Related Peptide in Primary Nephrotic Syndrome Patients

ＴｈｅｆｕｎｃｔｉｏｎｏｆｔｈｒｏｍｂｏｘａｎｅＢ2 （ＴＸＢ2 ）,6 ｋｅｔｏ ｐｒｏｓｔａｇｌａｎｄｉｎＦ1α （ 6 Ｋ ＰＧＦ1α）,ｅｎ ｄｏｔｈｅｌｉｎ（ＥＴ）ａｎｄｃａｌｃｉｔｏｎｉｎｇｅｎｅｒｅｌａｔｅｄｐｅｐ ｔｉｄｅ（ＣＧＲＰ）ｉｎｇｌｏｍｅｒｕｌｏｐａｔｈｙｈａｖｅａｌｒｅａｄｙａｒｏｕｓｅｄｉｎｔｅｒｓｔｏｆｃｌｉｎｉｃｉａｎｓ．（1 - 3） Ｓｉｎｃｅ 1 995,ｗｅｕｓｅｄＢｕｙａｎｇＨｕａｎｗｕＤｅｃｏｃｔｉｏｎ（补阳还五汤,ＢＹＨＷＤ）ａｓａｎｔｉｃｏａｇｕｌａｎｔｉｎｔｒｅａｔｉｎｇ…     

弓形虫主要表面抗原基因克隆及在大肠杆菌中的表达

为在大肠菌中表达弓形虫主要表面抗原（ＳＡＧ１）,进一步研究其生物学功能。将ＳＡＧ１基因重组于ｐＧＥＭＥＸ－１事例型表达载体上,转化大肠杆菌ＪＭ１０９（ＤＥ３）,得到含重组表达质粒ｐＧＥＭＥＸ－１／ＳＡＧ１的工程菌。结果表明ＩＰＴＧ诱导表达后,经ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ－ｂｌｏｔ检测证实含有ＳＡＧ１蛋白。提示ｐＧＥＭＥＸ－１大肠杆菌系统可以表达具有免疫活性的ＳＡＧ１蛋白。     

CORRELATION BETWEEN THE NUMBER OF MITOTIC FIGURES AND THE PERCENTAGE OF Ki67-POSITIVE CELLS IN NON-HODGKIN’S LYMPHOMAS

ＣＯＲＲＥＬＡＴＩＯＮＢＥＴＷＥＥＮＴＨＥＮＵＭＢＥＲＯＦＭＩＴＯＴＩＣＦＩＧＵＲＥＳＡＮＤＴＨＥＰＥＲＣＥＮＴＡＧＥＯＦＫｉ６７－ＰＯＳＩＴＩＶＥＣＥＬＬＳ　ＩＮＮＯＮ－ＨＯＤＧＫＩＮ’ＳＬＹＭＰＨＯＭＡＳＨｕａｎｇＧａｏｓｈｅｎｇ；（黄高升）Ｆｅ...     

Oxidatively Modified Very Low Density Lipoprotein Enhances Monocyte Adhesion to Endothelial Cells

ＯｘｉｄａｔｉｖｌｙＭｏｄｉｆｉｅｄＶｅｒｙＬｏｗＤｅｎｓｉｔｙＬｉｐｏｐｒｏｔｅｉｎＥｎｈａｎｃｅｓＭｏｎｏｃｙｔｅＡｄｈｅｓｉｏｎｔｏＥｎｄｏｔｈｅｌｉａｌＣｅｌｌｓＦＥＮＧＹｏｕ－ｍｅｉ（冯友梅）；ＺＨＡＮＧＺｈｉ－ｂｉｎｇ（张志兵）；ＷＡＮＧ...     

日本血吸虫重组BCG-Sj26GST疫苗对小鼠免疫应答的影响

采用日本血吸虫重组ＢＣＧ－Ｓｊ２６ＧＳＴ疫苗免疫ＢＡＬＢ／ｃ小鼠。免疫８周时,断头取血,取脾脏,取腹腔巨噬细胞。以淋巴细胞刺激指数（ＳＩ）反映细胞增殖能力,以ＮＯ释放量检测巨噬细胞吞噬活性,以ＥＬＩＳＡ试剂盒检测血清及脾淋巴细胞培养上清的白细胞介素２（ＩＬ－２）和干扰素－γ（ＩＦＮ－γ）的含量。结果表明,ｒＢＣＧ－Ｓｊ２６ＧＳＴ疫苗以 １０~６ＣＦＵ剂量经皮下免疫小鼠后,小鼠脾淋巴细胞增殖能力明显高于对照组、载体组和ＢＣＧ组（均为Ｐ＜０．０５）;小鼠腹腔巨噬细胞ＮＯ的含量明显高于对照组和载体组（均为Ｐ＜０．０１）;小鼠血清ＩＬ－２的含量明显高于对照组（Ｐ＜０．０１）和载体组（Ｐ＜０．０５）;而小鼠脾淋巴细胞培养上清ＩＬ－２的含量分别高于对照组４４％、载体组４８％和ＢＣＢ组４２％;小鼠血清ＩＦＮ－γ的含量分别高于对照组２０％、载体组１９％和ＢＣＧ组１３％,均有升高趋势、结果提示,ｒＢＣＧ－Ｓｊ２６ＧＳＴ疫苗能明显增强小鼠的免疫应答反应,其抗感染作用一方面与ＢＣＧ本身的佐剂特性有关,另一方面与机体产生抗Ｓｊ２６－ＧＳＴ特异性抗体有关。     

日本血吸虫（中国大陆株）Sj16基因的体外扩增及克隆

为了构建日本血吸虫（中国大陆株)Sj16基因原核表达重组质粒pGEX-4T-1-Sj16。根据曼氏血吸虫Sm16基因已知序列设计合成一对引物,用PCR技术从日本血吸虫（中国大陆株）成虫cDNA库中扩增Sj16基因;将Sj16基因定向克隆入pGEX-4T-1;转化感受态BL21/DE3菌;用酶切,PCR扩增鉴定筛选得到的重组阳性克隆。结果表明,从日本血吸虫（中国大陆株）成虫cDNA库中获取Sj16基因,重组质粒中含有Sj16基因。结果提示,成功构建日本血吸虫（中国大陆株）Sj16基因原核表达重组质粒pGEX-4T-1-Sj16,为进一步研究Sj16基因功能打下基础。     

Immune responses against Schistosoma japonicum after vaccinating mice with a multivalent DNA vaccine encoding integrated membrane protein Sj23 and cytokine interleukin-12

Background The vaccination of mice with DNA encoding single candidate antigens has failed to induce significant protection against Schistosoma japonicum ( S. japonicum) challenge infections. In this study, we evaluated the feasibility of using a multivalent DNA vaccine which co-expressed S.japonicum integral membrane protein Sj23 and murine cytokine IL-12 to induce protective immune responses.Methods The plasmid pVIVO2-IL12-Sj23, a eukaryotic expression vector expressing Sj23 and murine IL-12 simultaneously, was constructed, identified, and tested for expression in vitro. Its ability to protect against S. japonicum challenge infections was analyed according to worm reduction rate and egg reduction rate after vaccination of BALB/c mice. The serum levels of specific IgG antibody were determined by enzyme-linked-immuno sorbent assay (ELISA) and Western blot analysis. Using cultured spleen cells, IFN-γ and IL-4 post-stimulation were quantified by ELISA. The phenotypes of splenocyte populations were analyzed by flow cytometry (FCM).Results The plasmid DNA pVIVO2-IL12-Sj23 was proven to express well in vitro by transient transfection of HEK-293 cells. Immunization resulted in a worm reduction rate of 45. 53% and egg reduction rate of 58.35%. ELISA and Western blot analysis indicated that immunized mice generated specific IgG against Sj23. Spleen cells showed significant increases in IFN-γ but decreases in IL-4.No significant differences in CD4^ and CD8^ subgroup ratios were observed after the challenges.Conclusions The multivalent DNA vaccine pVIVO2-1L12-Sj23 is sufficient to elicit moderate but highly significant levels of protective immunity against challenge infections. Cytokine IL-12, as a gene adjuvant, was able to enhance the Thl responses and, hence, the protective immunity.     

rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值研究

目的研究rSj26-Sj32-Immunogold-Dipstick试剂对慢性日本血吸虫病的诊断价值。方法用rSj26-Sj32融合蛋白和日本血吸虫成虫抗原(SjAWA)Immunogold-Dipstick法检测慢性日本血吸虫病患者血清,同时以华支睾吸虫病、卫氏并殖吸虫病、泡型棘球蚴病、囊性棘球蚴病、乙型肝炎、肺结核患者和健康人血清作为对照,比较两种抗原检测抗体效果的差异。结果该法检测慢性日本血吸虫病的敏感性和特异性分别为92.50%和97.67%,诊断该病的阳性预告值、阴性预告值及诊断效率分别为97.37%、93.33%和95.18%,并且与其他疾病患者血清均无交叉反应。结论 rSj26-Sj32-Immunogold-Dipstick试剂可用于慢性日本血吸虫病的免疫诊断。     

重组核酸疫苗诱导小鼠抗血吸虫感染的免疫学研究

目的为日本血吸虫病的防治寻求新的高效联合基因免疫策略。方法构建共表达日本血吸虫相对分子质量23000表膜蛋白(Sj23)基因与鼠IL12基因的核酸疫苗质粒pVIVO2IL12Sj23,转染HEK293细胞,通过RTPCR,Westernblot及ELISA法检测Sj23和IL12蛋白的表达。接种并攻击感染BALB/c小鼠,以减虫率和减卵率评价其免疫保护力,同时设攻击感染对照组、空白质粒pVIVO2组、pVIVO2IL12组和pVIVO2Sj23组。用ELISA法和WesternBlot分析免疫小鼠血清中抗体。以脾细胞培养法检测经SEA刺激后,小鼠脾细胞分泌IFNγ和IL4的水平。并以FCM分析脾细胞亚群。结果经瞬时转染HEK293细胞证明质粒pVIVO2IL12Sj23能在体外进行表达。免疫小鼠后获得了4553%的减虫率和5835%的减卵率,较单价DNA疫苗pVIVO2Sj23免疫效果好(P<005)。ELISA法和WesternBlot检测抗体结果表明免疫小鼠产生了抗Sj23特异性IgG抗体。pVIVO2IL12Sj23免疫组IFNγ的水平较对照组显著升高而IL4水平较对照组低。攻击感染后各实验组小鼠脾细胞的CD4+,CD8+亚群比率无显著差别(P>005)。结论pVIVO2IL12Sj23DNA疫苗具有诱导BALB/c小鼠产生较好的抗血吸虫感染免疫保护效果。细胞因子IL12作为基因佐剂,具有诱导Th1型免疫反应从而增强DNA疫苗免疫保护性的作用。     

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjc14FABP and Sjc26GST genes and identify their expression in vitro, Sj 14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human plaeental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sl14 and Sl26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj 14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.     

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Gene

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjc14FABP and Sjc26GST genes and identify their expression in vitro, Sj 14 and Sj26 genes were ob- tained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sjl4 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 （IL-2） upstream Sj 14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyi-terminal of human placental alkaline phosphatase （PLAP） downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid plRES, resulting in another new recombinant plasmid plRES-Sj26-Sj14. The expression of Sj 14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays （IFA） when the plasmid plRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj 14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj 14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.     

日本血吸虫多价DNA疫苗pBK-Sj26(Sj32)-Sj23免疫效果的观察

为了观察血吸虫病多价DNA疫苗的保护力,将小鼠分成5组空白对照组、空质粒对照组、单价抗原DNA疫苗pBK-CMV-Sj23组、多价抗原DNA疫苗pBK-CMV-Sj26-Sj23和pBK-CMV-Sj32-Sj23组.大量提取各组质粒DNA后,各组于0、3、5周在BALB/c小鼠股四头肌注射相应质粒DNA,9周用血吸虫尾蚴攻击感染,15周剖杀小鼠计算减虫率及减卵率.结果显示与对照组比较,实验组小鼠减虫率及减卵率有极显著性差异(P<0.01);与单价pBK-CMV-Sj23组比较,多价DNA疫苗组的减虫率及减卵率有显著性差异.提示血吸虫多价DNA疫苗诱导小鼠对血吸虫的保护力优于单价DNA疫苗.     

Construction and Expression of DNA Vaccine pIRES-Sj97-Sj14-Sj26 and Its Immunogenicity in Mice

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj 14-Sj26 that contains fatty binding protein （Sj 14）, GST （Sj26） and paramyocin （Sj97） that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj 14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and plRES-Sj97-SjI4-Sj26 plasmid DNA, plRES-Sj 14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA, plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the plRES-Sj97-Sj 14-Sj26 group as compared with the plRES blank vector, normal saline and plRES-Sj26 groups （P〈 0.01） and the plRES-Sj 14-Sj26（P〈0.05）. Single splenocyte suspension was prepared to detected the level of IFN-T by ELISA and the lymphocyte stimulating index （SI） by MTT. SI was significantly higher of in the plRES-Sj97-Sj 14-Sj26 group than in other groups （P〈 0.01）, while the IFN-T level was significantly higher the plRES-Sj97-Sj 14-Sj26 group than in plRES blank vector and normal saline groups （P〈0.01）, but no significant differences were found when compared with plRES-Sj 14-Sj26 and plRES-Sj26 groups. Flow cytometery showed that the percentages of CD4＋ and CD8＋ T cells were much higher in the plRES-Sj97-Sj 14-Sj26 group （P〈 0.01, P〈0.05）. It was concluded that plRES-Sj97-Sj 14-Sj26 vaccine may induce stronger immune response in BALB/c mice.     

日本血吸虫新基因—腺苷酸激酶基因的发现与克隆

目的：将用表达序列标签（expression sequence tag,EST)策略及同源性搜索发现的日本血吸虫新基因－腺苷酸激酶（adenylate kinase,AK)cDNA克隆到表达质粒，pET32a( )上，为下一步研究该基因的功能做准备，方法：将插入于pTriplEx2质粒上的cDNA进行,测序，用BLASTn程序搜索测序结果，根据表达质粒pET32a( )上的克隆位点及该cDNA序列设计PCR引物，将PCR产物纯化后连接到pMD 18-T载体上，将重组T载体经EcoR I/Xho I双酶切后切下的SjAK基因导入原核一表达质粒pET32a( )中，结果：本研究所发现的新基因与曼氏血吸虫AK基因的同一性达86％，PCR产物的片段长度与预期大小一致，重组T载体及表达质粒经EcoR I及Xho I双酶切后有具有与目标片段工度相符的插入片段，结论：发现日本血吸虫的cDNA与曼氏血吸虫AKcDNA高度同源，并且成功地构建出重组表达质粒pET32a( )-SjAK。     

Construction and expression of DNA vaccine pIRES-Sj97-Sj14-Sj26 and its immunogenicity in mice

Summary To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, pIRES-Sj26 plasmid DNA, pIRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh. Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the pIRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT. SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P< 0.01, P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice. This project was supported by grants from National Natural Sciences Foundation of China (No. 30471603).