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Rapid,specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene

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Acta Oceanologica Sinica 2018年第4期37卷 83-88页

作者： ZHANG Lina WANG Mingyi CONG Dianxia DING Shuyan CONG Rinan YUE Jinyong GENG Jianli hu chengjin Department of Laboratory Medicine General Hospital of Jinan Military Region of PLA Department of Central Laboratory Weihai Municipal Hospital affiliated to Dalian Medical University

Vibrio vulnificus is an estuarine bacterial pathogen for *** rapid,specific and sensitive detection of *** is urgently needed for early disease diagnosis and timely treatment of *** *** the study,a loop-mediated isothermal amplification（LAMP） technique was developed for *** detection with a set of primers,composed of two out primers and two inner primers targeted to vvh A *** optimal amplification temperature was 63°C and the reaction only took 35 *** amplification products could not only be detected by agarose gel electrophoresis with ladder-like pattern bands,but also could be visualized using calcein with naked eye ***-five strains were tested for the specificity of LAMP assay,and all the *** strains were identified correctly while other strains were negative *** sensitive of the new LAMP assay was 100-fold more sensitive than the conventional ***,all the *** strains were detected correctly in spiked,clinical and environmental samples by the new LAMP *** with other well-known techniques,the new LAMP assay targeted to vvh A gene was extremely rapid,simple,sensitive and specific for *** identification.

关键词： Vibrio uulnificus loop-mediated isothermal amplification vvhA gene

Development and evaluation of an ELISA method for the measurement of kallikrein-related peptidase 5 (KLK5) in human serum

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Open Journal of Clinical Diagnostics 2013年第4期3卷 159-166页

作者： Yanhua Wu Xiaofei Liu Yingjian Chen chengjin hu Department of Laboratory Medicine Jinan General Hospital of PLA Jinan China

Kallikrein-related peptidases (KLKs) have been proposed as potential cancer biomarkers. Kallikrein-related peptidase 5 (KLK5) is a secreted trypsin-like protease of the KLKs. Until now, detection of KLK5 in both biological fluids and tissues has been described frequently due to the potential of being a new cancer biomarker. Our objective was to prepare KLK5 antibodies and establish an ELISA method for KLK5 to study the possible clinical application of KLK5 as a biomarker for malignancies. In this study, recombinant KLK5 protein was produced and purified using a prokaryotic expression system, and then used as immunogen to generate antibodies. High titers of specific antibodies were measured in serum of rabbits after the forth booster injection. And the titer of the antiserum reached 1:106. We have also generated monoclonal antibodies using hybridoma technology and the titer reached 1:105. The activity of KLK5 antibodies was characterized by Western blot and immunohistochemistry. To quantitatively examine KLK5 in serum samples, we established double antibody sandwich ELISA method using mouse mAb as capture and rabbit pAb as tracer antibody. We have detected KLK5 levels in ovarian cancer serum to ensure that our sandwich ELISA measurement to have high sensitivity and specificity. The ranges of linearity reached by the standard curves of the newly developed ELISA were 0.45 ng/mL to 125 ng/mL. The detection limit of the method, defined as the concentration of KLK5 can be distinguished, was 0.20 ng/mL. Median serum KLK5 levels were 3.77 ng/mL and 0.86 ng/mL in ovarian cancer patients and normal female, respectively (P ELISA assay for KLK5. Our preliminary findings prompt that KLK5 may be a new potential biomarker for the diagnosis and prognosis in patients with ovarian.

关键词： human Kallikrein 5 Kallikrein-Related Peptidases Generation of Antibodies Sandwich ELISA

The relationship between the expression of hK6 in ovarian neoplasm and the clinicopathologic variables and prognosis

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The Chinese-German Journal of Clinical Oncology 2010年第1期9卷 40-43页

作者： chengjin hu Fang Zhang Kuixiang Chen Yingjian Chen Department of Laboratory Medicine General Hospital of Jinan Military Region Jinan 250031 China Department of Pathology Zhejiang Cancer Hospital Zhejiang 310022 China

Objective: The aim of this study was to approach the relationship between the expression of human kallikrein 6 (hK6) in ovarian neoplasm and the clinicopathologic variables and prognosis for finding a new tumor marker for ovarian cancer. Methods: Through immunohistochemistry to examine the expression of hK6 in 19 cases with benign, 11 cases with borderline and 45 cases with malignant ovarian neoplasms and statistically analyzed whether the expression of hK6 correlated with the clinicopathologic variables and prognosis in patients with ovarian cancer. Results: The positive rate of hK6 in ovarian cancer tissues (60.0%) was significantly higher than that in the benign (15.8%) and borderline (27.3%) ovarian neoplasm tissues (P < 0.01). The expression of hK6 in low-grade ovarian cancer tissues was higher than that in high-grade (68.4% vs 14.3%; P < 0.05); hK6 in late–stage (stage III) was more frequently expressed than that in early-stage (stage I or II) (76.7% vs 26.7%; P < 0.01); and it significantly higher in ovarian carcinomas with lymph node metastases than those without lymph node metastases (77.8% vs 33.3%; P < 0.01 ); moreover, the expression of hK6 in the cancer tissues that the patients died or their pathogenetic condition recurred or their tumor metastasized within 3 years after surgery was higher (75.0%) than that in the cancer tissues that the pathogenetic condition of the patients was stable (42.9%; P < 0.05). Conclusion: The expression of hK6 in ovarian cancer tissues was higher than that in the benign and borderline ovarian neoplasm tissues, and high hK6 expression correlated with late–stage, low-grade, node metastasis and poor prognosis of patients. hK6 could potentially be a novel tumor marker for ovarian cancer that can predict the prognosis of the patients.

关键词： ovarian neoplasms human kallikrein 6 (hK6) immunohistochemistry tumor marker

急性白血病红细胞铜.锌—超氧化物歧化酶及血...

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中华血液学杂志 1992年第10期13卷 528-530页

作者：胡成进黄宁济南济南军区总医院检验科 250031 济南济南军区总医院检验科 250031；济南济南军区总医院临床血液科250031

测定了51例急性白血病患者在发病初期、缓解期及未缓解期红细胞Cu•Zn-超氧化物歧化酶（SOD）活性、血清Cu、Z n 及Cu/Zn比值，并与60例正常人进行对比分析，发现红细胞Cu • Z n -SOD活性在发病初期组明显下降（ P 0.05),并与红、白细胞... 详细信息

测定了51例急性白血病患者在发病初期、缓解期及未缓解期红细胞Cu•Zn-超氧化物歧化酶（SOD）活性、血清Cu、Z n 及Cu/Zn比值，并与60例正常人进行对比分析，发现红细胞Cu • Z n -SOD活性在发病初期组明显下降（ P 0.05),并与红、白细胞数目有关；血清Cu在发病初期组明显升高，Zn明显降低及Cu/Z n 比值增大（ P 0.05)，而Z n 及Cu/Zn比值则仍有显著性差异（未缓解组 P <0.01，缓解期组 P <0.05）。

关键词：白血病微量元素超氧物歧化酶

血清miRNAs和低剂量CT鉴别肺结节性质效果的研究

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中华肿瘤防治杂志 2024年第9期31卷 543-548页

作者：魏欢欢陈英剑胡成进山东第二医科大学医学检验学院山东潍坊261053 中国人民解放军联勤保障部队第九六〇医院检验科山东济南250031

目的研究miR-210、miR-1290和miR-19a在肺结节患者血清中的表达差异以及联合低剂量CT(LDCT)提高鉴别良恶性肺结节诊断效能的可行性。方法随机纳入2022-09-01-2023-03-31就诊于中国人民解放军联勤保障部队第九六〇医院的77例肺结节患者... 详细信息

目的研究miR-210、miR-1290和miR-19a在肺结节患者血清中的表达差异以及联合低剂量CT(LDCT)提高鉴别良恶性肺结节诊断效能的可行性。方法随机纳入2022-09-01-2023-03-31就诊于中国人民解放军联勤保障部队第九六〇医院的77例肺结节患者为研究对象,所有患者均进行LDCT检查,并采用实时荧光定量聚合酶链式反应(qRT-PCR)检测患者血清miR-210、miR-1290和miR-19a的相对表达水平以鉴别肺结节性质,手术或穿刺活检的病理诊断结果为金标准。采用χ^(2)检验分析良恶性肺结节与影像学征象的关系,Mann-Whitney U检验进行组间各miRNAs相对表达的比较,通过绘制ROC曲线,评价3种miRNAs鉴别良恶性肺结节的效果,进一步评价miRNAs联合LDCT扫描结果的鉴别诊断效果。结果LDCT肺扫描检出恶性肺结节61例,良性肺结节16例。miR-210、miR-1290和miR-19a在恶性肺结节患者血清中的表达均高于良性肺结节患者,均P<0.05。ROC曲线显示,miR-210、miR-1290和miR-19a诊断良恶性肺结节的AUC值分别为0.699、0.709和0.697,3种miRNAs联合检测的AUC值为0.768。3种miRNAs联合检测检出恶性肺结节37例,良性肺结节40例。通过病理学组织切片确诊恶性肺结节47例、良性肺结节30例。其中LDCT检出的41例恶性肺结节和10例良性肺结节与病理学诊断结果一致,鉴别肺结节性质的准确率为66.2%。影像学显示,恶性肺结节者的结节密度改变、分叶征、毛刺征、空泡征、胸膜牵拉发生率显著高于良性肺结节者,χ^(2)值分别为13.508、10.100、6.337、18.731和13.766,均P<0.05。3种miRNAs联合检测检出的30例恶性肺结节和23例良性肺结节与病理学诊断结果一致,鉴别肺结节性质的准确率为68.8%。LDCT与3种miRNAs联合检测后,检出的44例恶性肺结节和26例良性肺结节与病理学诊断结果一致,鉴别肺结节性质的灵敏度、特异度和准确率分别为93.6%、86.7%和90.9%。结论LDCT肺扫描联合血清miR-210、miR-1290和miR-19a检测能提高鉴别肺结节性质的准确性,对肺结节良恶性的鉴别诊断具有实际临床意义。

关键词：肺结节 miRNA 低剂量CT 鉴别诊断

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阴沟肠杆菌的临床分布及耐药性变迁

中国感染与化疗杂志 2015年第5期15卷 459-461页

作者：卢兆莲公衍文薛炼胡成进济南军区总医院实验诊断科济南250031

目的分析阴沟肠杆菌临床分离株的分布特点,以及对抗菌药物的耐药性变迁,为临床合理使用抗菌药物提供依据。方法收集2009年1月—2013年12月济南军区总医院患者各类标本分离的阴沟肠杆菌,使用VITEK-2 Compact全自动细菌分析仪对所分离的... 详细信息

目的分析阴沟肠杆菌临床分离株的分布特点,以及对抗菌药物的耐药性变迁,为临床合理使用抗菌药物提供依据。方法收集2009年1月—2013年12月济南军区总医院患者各类标本分离的阴沟肠杆菌,使用VITEK-2 Compact全自动细菌分析仪对所分离的菌株进行鉴定;采用K-B纸片扩散法进行药敏试验,并对结果进行分析。结果 5年非重复分离的阴沟肠杆菌397株,约占所有非重复分离革兰阴性菌株的5.1%,5年中分离率变化不大;主要分离自痰液、分泌物、尿液,分别占48.9%、30.5%、7.3%。除对碳青霉烯类、头孢西丁和阿莫西林-克拉维酸外,阴沟肠杆菌的总体耐药率呈下降趋势。但碳青霉烯类抗生素对阴沟肠杆菌有较好的抗菌活性。该菌对β内酰胺类抗生素耐药率较高。结论应加强医院临床分离菌的耐药性监测,指导临床合理选用抗菌药物。

关键词：阴沟肠杆菌细菌分布细菌耐药性

血清甘氨酰脯氨酸二肽氨基肽酶对原发性肝癌的诊断价值探讨

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山东医药 2000年第17期40卷 11-12页

作者：牛爱军胡成进闫先侠济南军区总医院山东济南250031 山东省精神卫生中心

对 31例原发性肝癌 (PHC)、 32例肝硬化患者及 2 0例正常人进行了血清甘氨酰脯氨酸二肽氨基肽酶(GPDA)的活性测定 ,对 31例 PHC患者治疗前后 GPDA的含量进行了对比分析 ,并与甲胎蛋白 (AFP)的诊断价值进行对比。结果显示 ,GPDA对 PHC的... 详细信息

对 31例原发性肝癌 (PHC)、 32例肝硬化患者及 2 0例正常人进行了血清甘氨酰脯氨酸二肽氨基肽酶(GPDA)的活性测定 ,对 31例 PHC患者治疗前后 GPDA的含量进行了对比分析 ,并与甲胎蛋白 (AFP)的诊断价值进行对比。结果显示 ,GPDA对 PHC的诊断敏感性为 74.2 % ,优于 AFP诊断敏感性 6 7.7% ,特别是在 AFP阴性的 PHC患者中 ,GPDA的阳性率高达 6 0 % ,弥补了 AFP对

关键词：甘氨酰脯氨酸二肽氨基肽酶原发性肝癌甲胎蛋白

肠道微生态影响肿瘤免疫检查点抑制剂疗效的研究进展

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中华检验医学杂志 2019年第5期42卷 386-389页

作者：周亚男张晓彤胡成进曹源解放军第九六〇医院实验诊断科济南250031

免疫检查点抑制剂(ICIs)逐渐成为治疗恶性肿瘤的新策略,多个药物已被批准上市,但是只有10%~30%的患者受益。研究表明,肠道微生态可通过免疫调节来影响肿瘤患者ICIs的治疗效果。本文回顾了近年来肠道微生态对ICIs治疗的影响及调节作用,... 详细信息

免疫检查点抑制剂(ICIs)逐渐成为治疗恶性肿瘤的新策略,多个药物已被批准上市,但是只有10%~30%的患者受益。研究表明,肠道微生态可通过免疫调节来影响肿瘤患者ICIs的治疗效果。本文回顾了近年来肠道微生态对ICIs治疗的影响及调节作用,介绍肠道微生态影响免疫治疗的可能机制,总结肠道微生态相关检测方法,对其临床应用转化的前景进行展望。

关键词：胃肠道微生物组癌,非小细胞肺黑色素瘤

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微生物群定植抵抗力抗感染研究进展

实用医药杂志 2021年第7期38卷 647-650页

作者：闵彦胡成进解放军第960医院山东济南250031

近年来,微生物群定植抵抗力抗感染能力越来越引起研究人员的注意。该文将微生物群对人体的保护作用,微生物群对耐药菌的拮抗作用以及其中所涉及的细菌之间的竞争机制做一综述,以期为抗生素合理使用及耐药菌感染防治提供新的理论依据。

关键词：微生物群定植抵抗力感染耐药菌