目的观察血清成纤维生长因子-23(fibroblast growth factor 23,FGF23)/抗衰老基因Klotho、辅助性T淋巴细胞(helper T lymphocytes,Th)1/Th2细胞因子在老年终末期肾病患者中的表达,并分析对医院感染的预测和预后的影响。方法选取2021年1...目的观察血清成纤维生长因子-23(fibroblast growth factor 23,FGF23)/抗衰老基因Klotho、辅助性T淋巴细胞(helper T lymphocytes,Th)1/Th2细胞因子在老年终末期肾病患者中的表达,并分析对医院感染的预测和预后的影响。方法选取2021年1月1日至2022年12月31日自贡市精神卫生中心(自贡市老年病医院)收治的106例老年终末期肾病患者,男65例,年龄范围60~84岁,年龄(67.21±3.07)岁,女41例,年龄范围60~84岁,年龄(67.65±2.98)岁,于入院第2天检测血清FGF23、抗衰老基因Klotho、Th1细胞因子[γ干扰素(interferon-γ,IFN-γ)、白介素2(interleukin-2,IL-2)]、Th2细胞因子(IL-4、IL-10)水平,进行急性生理功能及慢性健康状况评分Ⅱ(acute physiology and chronic health status scoring systemⅡ,APACHEⅡ)评估。根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),比较两组各项血清指标及APACHEⅡ评分水平,分析各项指标与APACHEⅡ评分的相关性。随访3个月统计患者生存预后[生存(71例)、死亡(35例)],比较不同预后患者血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10水平,分析各项指标对医院感染及预后的预测价值及临床效用。结果106例老年终末期肾病患者根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),随访3个月后生存71例、死亡35例。入院第2天感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分分别为(78.64±20.16)ng/mL、(20.14±1.48)μg/L、(22.47±2.56)μg/L、(26.38±6.51)分,未感染组分别为(60.17±16.83)ng/mL、(16.25±1.21)μg/L、(19.52±1.86)μg/L、(22.97±6.45)分,感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分均高于未感染组(P<0.05);入院第2天感染组血清Klotho、IFN-γ、IL-2水平分别为(34.95±12.62)ng/mL、(22.19±1.69)μg/L、(28.73±2.95)μg/L,未感染组分别为(51.61±16.08)ng/mL、(25.31±1.74)μg/L、(33.95±1.52)μg/L,感染组血清Klotho、IFN-γ、IL-2水平均低于未感染组(P<0.05);血清FGF23、IL-4、IL-10水平与APACHEⅡ评分呈正相关(相关系数r=0.629、0.597、0.612,P均<0.05),血清Klotho、IFN-γ、IL-2水平与APACHEⅡ评分呈负相关(相关系数r=-0.632、-0.718、-0.701、0.597,P均<0.05);死亡患者入组后1个月血清FGF23、IL-10、IL-4水平分别为(77.49±21.85)ng/mL、(24.76±4.77)μg/L、(24.81±6.28)μg/L,生存患者分别为(58.92±16.94)ng/mL、(18.10±3.82)μg/L、(13.57±4.38)μg/L,死亡患者入组后1个月血清FGF23、IL-10、IL-4水平高于生存患者(P<0.05);死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平分别为(30.03±11.76)ng/mL、(20.33±2.63)μg/L、(27.19±4.91)μg/L,生存患者分别为(55.68±17.02)ng/mL、(26.54±4.79)μg/L、(35.22±5.64)μg/L,死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平低于生存患者(P<0.05);血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10预测终末期老年肾病患者发生医院感染、生存预后曲线下面积分别为0.904(OR:0.832~0.953)、0.911(OR:0.840~0.958),且具有良好临床效用(P<0.05)。结论老年终末期肾病患者血清FGF23/Klotho、Th1/Th2细胞因子水平失衡,合并感染患者血清FGF23、IL-10、IL-4水平异常升高,血清Klotho、IFN-γ、IL-2水平表达降低,联合血清FGF23/Klotho、Th1/Th2细胞因子预测医院感染、评估预后的价值较高。展开更多
[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its g...[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its genomic DNA by PCR amplification, and cloned into the pGEM-Easy vector; then the sequencing result was analyzed with bioinformatics methods. [Result] Whole length of the P23 gene from Theileria sergenti is 684 bp containing a 672 bp open reading frame. The deduced amino acid sequence (223 amino acid residues) contains a signal peptide of 19 amino acid residues and two fragments of transmembrane domains, with relative molecular weight of the 25.886 kD and with the pI of 9.22. The homology between the yielded sequence and Chitose of Theileria sergenti P23 gene(TS-Chitose type, D84446), Ikeda of Theileria sergenti P23 gene(TS-Ikeda type, D84447) reached 99% and 90%, respectively. The sequence has been accessed in GenBank(EU573168). [Conclusion] The protein encoded by the P23 gene has better stability and immunogenicity, thus can be used as the antigen candidate for preparing genetic engineering vaccine for Theileria sergenti.展开更多
[Objective] The aim was to study cloning and prokaryotic expression of P23 major surface protein gene of Theileria sergenti. [Method] A pair of specific primers was designed according to the sequence of P23 major surf...[Objective] The aim was to study cloning and prokaryotic expression of P23 major surface protein gene of Theileria sergenti. [Method] A pair of specific primers was designed according to the sequence of P23 major surface protein of T. sergenti (D84447).The P23 gene was amplified by PCR from genomic DNA of T. sergenti and cloned into pMD18-T vector to construct recombinant clonal vector pMD18-P23. Positive clones were identified by PCR screening and restriction digestion. A recombinant expression plasmid pGEX-4T-P23 was constructed by subcloning the cloned P23 gene into the linearized pGEX-4T-1 vector and transformed into E. coli BL21. After introduction by IPTG,the expressed fusion protein was identified by SDS-PAGE and Western-blotting. [Result] The cloned gene has a total length of 507 bp. Sequencing result showed that the nucleotide sequence of the cloned P23 gene shared 99.4% identity with that of P23 published in GenBank (D84447). The expressed fusion protein was 46 ku in molecular mass. Induction opportunity of zhours after culture inoculation was the best,the induction time of 6 h was the best,and induction temperature of 34 ℃ was the best as well,IPTG of 1 mmol/L had little effect on the expression. Western-blotting indicated that recombinant protein was recognized by specific antibody. [Conclusion] This study would lay a foundation for further research on the prevention and diagnose of T. sergenti.展开更多
nm-23 gene is the first antimetastatic gene which was found to have close relationship with metastasis and prognosis of many malignant carcinoma.This review briefly summarizes the development of nm-23 gene and hepatoc...nm-23 gene is the first antimetastatic gene which was found to have close relationship with metastasis and prognosis of many malignant carcinoma.This review briefly summarizes the development of nm-23 gene and hepatocellular carcinoma in recent years.展开更多
文摘目的观察血清成纤维生长因子-23(fibroblast growth factor 23,FGF23)/抗衰老基因Klotho、辅助性T淋巴细胞(helper T lymphocytes,Th)1/Th2细胞因子在老年终末期肾病患者中的表达,并分析对医院感染的预测和预后的影响。方法选取2021年1月1日至2022年12月31日自贡市精神卫生中心(自贡市老年病医院)收治的106例老年终末期肾病患者,男65例,年龄范围60~84岁,年龄(67.21±3.07)岁,女41例,年龄范围60~84岁,年龄(67.65±2.98)岁,于入院第2天检测血清FGF23、抗衰老基因Klotho、Th1细胞因子[γ干扰素(interferon-γ,IFN-γ)、白介素2(interleukin-2,IL-2)]、Th2细胞因子(IL-4、IL-10)水平,进行急性生理功能及慢性健康状况评分Ⅱ(acute physiology and chronic health status scoring systemⅡ,APACHEⅡ)评估。根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),比较两组各项血清指标及APACHEⅡ评分水平,分析各项指标与APACHEⅡ评分的相关性。随访3个月统计患者生存预后[生存(71例)、死亡(35例)],比较不同预后患者血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10水平,分析各项指标对医院感染及预后的预测价值及临床效用。结果106例老年终末期肾病患者根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),随访3个月后生存71例、死亡35例。入院第2天感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分分别为(78.64±20.16)ng/mL、(20.14±1.48)μg/L、(22.47±2.56)μg/L、(26.38±6.51)分,未感染组分别为(60.17±16.83)ng/mL、(16.25±1.21)μg/L、(19.52±1.86)μg/L、(22.97±6.45)分,感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分均高于未感染组(P<0.05);入院第2天感染组血清Klotho、IFN-γ、IL-2水平分别为(34.95±12.62)ng/mL、(22.19±1.69)μg/L、(28.73±2.95)μg/L,未感染组分别为(51.61±16.08)ng/mL、(25.31±1.74)μg/L、(33.95±1.52)μg/L,感染组血清Klotho、IFN-γ、IL-2水平均低于未感染组(P<0.05);血清FGF23、IL-4、IL-10水平与APACHEⅡ评分呈正相关(相关系数r=0.629、0.597、0.612,P均<0.05),血清Klotho、IFN-γ、IL-2水平与APACHEⅡ评分呈负相关(相关系数r=-0.632、-0.718、-0.701、0.597,P均<0.05);死亡患者入组后1个月血清FGF23、IL-10、IL-4水平分别为(77.49±21.85)ng/mL、(24.76±4.77)μg/L、(24.81±6.28)μg/L,生存患者分别为(58.92±16.94)ng/mL、(18.10±3.82)μg/L、(13.57±4.38)μg/L,死亡患者入组后1个月血清FGF23、IL-10、IL-4水平高于生存患者(P<0.05);死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平分别为(30.03±11.76)ng/mL、(20.33±2.63)μg/L、(27.19±4.91)μg/L,生存患者分别为(55.68±17.02)ng/mL、(26.54±4.79)μg/L、(35.22±5.64)μg/L,死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平低于生存患者(P<0.05);血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10预测终末期老年肾病患者发生医院感染、生存预后曲线下面积分别为0.904(OR:0.832~0.953)、0.911(OR:0.840~0.958),且具有良好临床效用(P<0.05)。结论老年终末期肾病患者血清FGF23/Klotho、Th1/Th2细胞因子水平失衡,合并感染患者血清FGF23、IL-10、IL-4水平异常升高,血清Klotho、IFN-γ、IL-2水平表达降低,联合血清FGF23/Klotho、Th1/Th2细胞因子预测医院感染、评估预后的价值较高。
文摘[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its genomic DNA by PCR amplification, and cloned into the pGEM-Easy vector; then the sequencing result was analyzed with bioinformatics methods. [Result] Whole length of the P23 gene from Theileria sergenti is 684 bp containing a 672 bp open reading frame. The deduced amino acid sequence (223 amino acid residues) contains a signal peptide of 19 amino acid residues and two fragments of transmembrane domains, with relative molecular weight of the 25.886 kD and with the pI of 9.22. The homology between the yielded sequence and Chitose of Theileria sergenti P23 gene(TS-Chitose type, D84446), Ikeda of Theileria sergenti P23 gene(TS-Ikeda type, D84447) reached 99% and 90%, respectively. The sequence has been accessed in GenBank(EU573168). [Conclusion] The protein encoded by the P23 gene has better stability and immunogenicity, thus can be used as the antigen candidate for preparing genetic engineering vaccine for Theileria sergenti.
基金Supported by Science and Technology Development Program of Jilin Province (20050703-4)~~
文摘[Objective] The aim was to study cloning and prokaryotic expression of P23 major surface protein gene of Theileria sergenti. [Method] A pair of specific primers was designed according to the sequence of P23 major surface protein of T. sergenti (D84447).The P23 gene was amplified by PCR from genomic DNA of T. sergenti and cloned into pMD18-T vector to construct recombinant clonal vector pMD18-P23. Positive clones were identified by PCR screening and restriction digestion. A recombinant expression plasmid pGEX-4T-P23 was constructed by subcloning the cloned P23 gene into the linearized pGEX-4T-1 vector and transformed into E. coli BL21. After introduction by IPTG,the expressed fusion protein was identified by SDS-PAGE and Western-blotting. [Result] The cloned gene has a total length of 507 bp. Sequencing result showed that the nucleotide sequence of the cloned P23 gene shared 99.4% identity with that of P23 published in GenBank (D84447). The expressed fusion protein was 46 ku in molecular mass. Induction opportunity of zhours after culture inoculation was the best,the induction time of 6 h was the best,and induction temperature of 34 ℃ was the best as well,IPTG of 1 mmol/L had little effect on the expression. Western-blotting indicated that recombinant protein was recognized by specific antibody. [Conclusion] This study would lay a foundation for further research on the prevention and diagnose of T. sergenti.
文摘nm-23 gene is the first antimetastatic gene which was found to have close relationship with metastasis and prognosis of many malignant carcinoma.This review briefly summarizes the development of nm-23 gene and hepatocellular carcinoma in recent years.
