目的:探讨抗中性粒细胞胞质抗体谱(antineutrophil cytoplasmic antibodies,ANCAs)检测在自身免疫性肝炎(autoimmune hepatitis,AIH)患者的临床诊断价值,为AIH的相关研究提供临床依据.方法:对南昌大学第二附属医院2008-07/2012-03符合...目的:探讨抗中性粒细胞胞质抗体谱(antineutrophil cytoplasmic antibodies,ANCAs)检测在自身免疫性肝炎(autoimmune hepatitis,AIH)患者的临床诊断价值,为AIH的相关研究提供临床依据.方法:对南昌大学第二附属医院2008-07/2012-03符合纳入标准的48例AIH、190例非-AIH患者及50例健康体检者采用ELISA法检测抗乳铁蛋白抗体(anti-lactoferrin antibody,ALA)、抗蛋白酶3抗体(抗-PR3)和抗髓过氧化物酶抗体(抗-MPO),免疫荧光法检测ANCAs,回顾性分析检测结果.结果:(1)AIH组ANCA、p-ANCA、x-ANCA及ALA阳性率与非-AIH各组(PSC除外)比较有统计学意义,抗-MPO阳性率为54.2%,与非AIH各组比较均有P<0.01;(2)所有标本中仅AIH组有1例同时c-ANCA、抗-PR3阳性,虽对诊断AIH特异性高但无临床价值;(3)配对2检测,抗-MPO与ANCA、p-ANCA,ALA与x-A N C A对A I H的诊断无统计学意义;一致性分析,抗-MPO与ANCA、p-ANCA值为0.819、0.875,一致性极好.结论:ANCAs抗体间存在一定的相关性,联合检测可提高AIH的诊断率,对AIH患者的临床鉴别、诊断有重要意义.展开更多
AIM: To construct fusion protein of a single-chain antibody (scFv) against transferrin receptor (TfR) with alkaline phosphatase(AP). METHODS: The VH-linker-VL,namely scFv gene,was prepared by amplifying the VH and VL ...AIM: To construct fusion protein of a single-chain antibody (scFv) against transferrin receptor (TfR) with alkaline phosphatase(AP). METHODS: The VH-linker-VL,namely scFv gene,was prepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by 5/71 and Not I,it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E.coli TG1.The positive colonies were screened by colony PCR and their expressions were induced by IPTG.ScFv gene was gained by digesting ScFv expression vector pUC19/119 with 5/71 and NotI restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E.coli TG1.The positive colonies were selected by bacterial colony PCR.The expression of fusion protein (scFv-AP) was induced by IPTG.Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku.Immunofluorescent assay (IFA) demonstrated its reactivity with TfR.The molecular weight of scFv-AP was 75 ku.Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity. CONCLUSION: We have successfully prepared the anti-human TfR scFv and constructed the fusion protein of scFv and AP.It is promising for immunological experiments.展开更多
Iron deficiency （ID）, with or without anemia, is often caused by digestive diseases and should always be investigated, except in very specific situations, as its causes could be serious diseases, such as cancer. Dia...Iron deficiency （ID）, with or without anemia, is often caused by digestive diseases and should always be investigated, except in very specific situations, as its causes could be serious diseases, such as cancer. Diagnosis of ID is not always easy. Low serum levels of ferritin or transferrin saturation, imply a situation of absolute or functional ID. It is sometimes difficult to differentiate ID anemia from anemia of chronic diseases, which can coexist. In this case, other parameters, such as soluble transferrin receptor activity can be very useful. After an initial evaluation by clinical history, urine analysis, and serological tests for celiac disease, gastroscopy and colonoscopy are the key diagnostic tools for investigating the origin of ID, and will detect the most important and prevalent diseases. If both tests are normal and anemia is not severe, treatment with oral iron can be indicated, along with stopping any treatment with non-steroidal anti-inflammatory drugs. In the absence of response to oral iron, or if the anemia is severe or clinical suspicion of important disease persists, we must insist on diagnostic evaluation. Repeat endoscopic studies should be considered in many cases and if both still show normal results, investigating the small bowel must be considered. The main techniques in this case are capsule endoscopy, followed by展开更多
Objective： Iron deficiency is a common complication in patients with polycythemia vera （PV）. Hepcidin is a principal regulator of iron homeostasis. The aim of our study was to assess prohepcidin, a hepcidin precurs...Objective： Iron deficiency is a common complication in patients with polycythemia vera （PV）. Hepcidin is a principal regulator of iron homeostasis. The aim of our study was to assess prohepcidin, a hepcidin precursor, and other iron status pa- rameters in the serum of PV patients. Methods： The study was performed in 60 patients （F/M 26/34） aged 38-84 （66＋ 10） years. The control group consisted of 20 healthy volunteers, age and sex matched. The following parameters were determined in blood serum samples： prohepcidin concentration, iron content, unsaturated iron binding capacity （UIBC）, total iron binding capacity （TIBC）, transferrin saturation （TfS）, and concentrations of ferritin and soluble transferrin receptor （sTfR）. Results： All PV patients showed significantly lower levels of prohepcidin, higher levels of sTIR and TIBC compared to the control group. 40% of the patients from the study group showed concentrations of ferritin below the normal range and significantly lower levels of serum iron and TfS, and significantly higher levels of sTIR, UIBC and TIBC in comparison with the rest of the study group. In this group of patients, prohepcidin concentrations were significantly lower than those in other patients. Conclusion： The results indicate that PV patients suffer from iron metabolism disorders. The decreased serum level ofprohepcidin in PV patients may be a result of iron deficiency.展开更多
文摘目的:探讨抗中性粒细胞胞质抗体谱(antineutrophil cytoplasmic antibodies,ANCAs)检测在自身免疫性肝炎(autoimmune hepatitis,AIH)患者的临床诊断价值,为AIH的相关研究提供临床依据.方法:对南昌大学第二附属医院2008-07/2012-03符合纳入标准的48例AIH、190例非-AIH患者及50例健康体检者采用ELISA法检测抗乳铁蛋白抗体(anti-lactoferrin antibody,ALA)、抗蛋白酶3抗体(抗-PR3)和抗髓过氧化物酶抗体(抗-MPO),免疫荧光法检测ANCAs,回顾性分析检测结果.结果:(1)AIH组ANCA、p-ANCA、x-ANCA及ALA阳性率与非-AIH各组(PSC除外)比较有统计学意义,抗-MPO阳性率为54.2%,与非AIH各组比较均有P<0.01;(2)所有标本中仅AIH组有1例同时c-ANCA、抗-PR3阳性,虽对诊断AIH特异性高但无临床价值;(3)配对2检测,抗-MPO与ANCA、p-ANCA,ALA与x-A N C A对A I H的诊断无统计学意义;一致性分析,抗-MPO与ANCA、p-ANCA值为0.819、0.875,一致性极好.结论:ANCAs抗体间存在一定的相关性,联合检测可提高AIH的诊断率,对AIH患者的临床鉴别、诊断有重要意义.
文摘铁蛋白是肝癌、何杰金氏病、肺未分化癌,神经母细胞瘤、急性粒细胞性白血病、乳癌和胰癌的肿瘤相关蛋白质。静脉输入131I 标记异种抗铁蛋白IgG 抗体,通过扫描,可精确显示多种原发癌和转移癌的体内位置。根据这一特点,这类抗体应能用于肿瘤局部的体内选择性放疗。霍普金斯医院肿瘤中心,制备131I 标记抗铁蛋白IgG 的步骤为:由何杰金氏病病人的脾脏提取并纯化铁蛋白,加佛氏完全佐剂,重复接种于新西兰白兔,取得兔抗人铁蛋白免疫血清,通过沉淀、透析取得免疫IgG。按乳过氧化酶法每5mg IgG 加入25ug 乳过氧化酶,1mci Na131I,100 ul 8 m M H2O2。
基金Supported by Natural Key and Basic Research Development Program,No.2002CB513109
文摘AIM: To construct fusion protein of a single-chain antibody (scFv) against transferrin receptor (TfR) with alkaline phosphatase(AP). METHODS: The VH-linker-VL,namely scFv gene,was prepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by 5/71 and Not I,it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E.coli TG1.The positive colonies were screened by colony PCR and their expressions were induced by IPTG.ScFv gene was gained by digesting ScFv expression vector pUC19/119 with 5/71 and NotI restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E.coli TG1.The positive colonies were selected by bacterial colony PCR.The expression of fusion protein (scFv-AP) was induced by IPTG.Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku.Immunofluorescent assay (IFA) demonstrated its reactivity with TfR.The molecular weight of scFv-AP was 75 ku.Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity. CONCLUSION: We have successfully prepared the anti-human TfR scFv and constructed the fusion protein of scFv and AP.It is promising for immunological experiments.
文摘Iron deficiency （ID）, with or without anemia, is often caused by digestive diseases and should always be investigated, except in very specific situations, as its causes could be serious diseases, such as cancer. Diagnosis of ID is not always easy. Low serum levels of ferritin or transferrin saturation, imply a situation of absolute or functional ID. It is sometimes difficult to differentiate ID anemia from anemia of chronic diseases, which can coexist. In this case, other parameters, such as soluble transferrin receptor activity can be very useful. After an initial evaluation by clinical history, urine analysis, and serological tests for celiac disease, gastroscopy and colonoscopy are the key diagnostic tools for investigating the origin of ID, and will detect the most important and prevalent diseases. If both tests are normal and anemia is not severe, treatment with oral iron can be indicated, along with stopping any treatment with non-steroidal anti-inflammatory drugs. In the absence of response to oral iron, or if the anemia is severe or clinical suspicion of important disease persists, we must insist on diagnostic evaluation. Repeat endoscopic studies should be considered in many cases and if both still show normal results, investigating the small bowel must be considered. The main techniques in this case are capsule endoscopy, followed by
基金Project supported partly by the European Social Fund and the Polish Government within the Integrated Regional Development Operational Programme, the project "Scholarship for PhD Students 2008/2009-ZPORR" of Kuyavian-Pomeranian Voivodeshipa grant awarded by Nicolaus Copernicus University (No. 04/2008), Poland
文摘Objective： Iron deficiency is a common complication in patients with polycythemia vera （PV）. Hepcidin is a principal regulator of iron homeostasis. The aim of our study was to assess prohepcidin, a hepcidin precursor, and other iron status pa- rameters in the serum of PV patients. Methods： The study was performed in 60 patients （F/M 26/34） aged 38-84 （66＋ 10） years. The control group consisted of 20 healthy volunteers, age and sex matched. The following parameters were determined in blood serum samples： prohepcidin concentration, iron content, unsaturated iron binding capacity （UIBC）, total iron binding capacity （TIBC）, transferrin saturation （TfS）, and concentrations of ferritin and soluble transferrin receptor （sTfR）. Results： All PV patients showed significantly lower levels of prohepcidin, higher levels of sTIR and TIBC compared to the control group. 40% of the patients from the study group showed concentrations of ferritin below the normal range and significantly lower levels of serum iron and TfS, and significantly higher levels of sTIR, UIBC and TIBC in comparison with the rest of the study group. In this group of patients, prohepcidin concentrations were significantly lower than those in other patients. Conclusion： The results indicate that PV patients suffer from iron metabolism disorders. The decreased serum level ofprohepcidin in PV patients may be a result of iron deficiency.
