The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothe...The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothermal titration calorimetry(ITC) measurement. The results showed that there was competitive adsorption between heparin and fibronectin, and the preadsorption of fibronectin could prevent subsequent heparin adsorption to some extent, and the adsorbed Hep/Fn complex on the surface was in a rigid form. The bioactivity of heparin and fibronectin could be affected by the bulk concentration of each, and both heparin and fibronectin in Hep/Fn complex formed under p H 4 condition displayed larger bioactivity than that formed under p H 7 condition. Moreover, the fibronectin showed more exposed cell-binding sites at the p H value lower than physiological condition. The results of ITC further suggested that the interaction between heparin and fibronectin under p H 4 was stronger than under p H 7, and the complex was also more stable. The study brings forth the detailed interaction between heparin and fibronectin, which will be helpful for better understanding the interaction mechanism of the two biomolecules. The results may be potentially useful for the development of new generation of cardiovascular biomaterials.展开更多
BACKGROUND The microsatellite instability(MSI)test and immunohistochemistry(IHC)are widely used to screen DNA mismatch repair(MMR)deficiency in sporadic colorectal cancer(CRC).For IHC,a two-antibody panel of MLH1 and ...BACKGROUND The microsatellite instability(MSI)test and immunohistochemistry(IHC)are widely used to screen DNA mismatch repair(MMR)deficiency in sporadic colorectal cancer(CRC).For IHC,a two-antibody panel of MLH1 and MSH2 or four-antibody panel of MLH1,MSH2,PMS2,and MSH6 are used.In general,MSI is known as a more accurate screening test than IHC.AIM To compare two-and four-antibody panels of IHC in terms of accuracy and cost benefit on the basis of MSI testing for detecting MMR deficiency.METHODS We retrospectively analyzed patients with CRC who underwent curative surgery between 2015 and 2017 at a tertiary referral center.Both IHC with four antibodies and MSI tests were routinely performed.The sensitivity and specificity of a fourand two types of two-antibody panels(PMS2/MSH6 and MLH1/MSH2)were compared on the basis of MSI testing for detecting MMR deficiency.RESULTS High-frequency MSI was found in 5.5%(n=193)of the patients(n=3486).The sensitivities of the four-and two types of two-antibody panels were 97.4%,92.2%,and 87.6%,respectively.The specificities of the three types of panels did not differ significantly(99.6%for the four-antibody and PMS2/MSH6 panels,99.7%for the MLH1/MSH2 panel).Based on Cohen's kappa statistic(κ),four-and twoantibody panels were in almost perfect agreement with the MSI test(κ>0.9).The costs of the MSI test and the four-and two-antibody panels of IHC were approximately$200,$160,and$80,respectively.CONCLUSION Considering the cost of the four-antibody panel IHC compared to that of the twoantibody panel IHC,a two-antibody panel of PMS2/MSH6 might be the best choice in terms of balancing cost-effectiveness and accuracy.展开更多
Immunocytochemical localization of 5-hydroxytryptamine (5-HT) containing cells of the guinea-pig pancreas was studied by means of PAP-glucose oxidase-DAB-nickel (PAP-GDN) and PAPstaining methods on paraffin sections. ...Immunocytochemical localization of 5-hydroxytryptamine (5-HT) containing cells of the guinea-pig pancreas was studied by means of PAP-glucose oxidase-DAB-nickel (PAP-GDN) and PAPstaining methods on paraffin sections. In this study we demonstrated the presence of 5-HTimmunoreactive cells in the endocrine (islets) and exocrine pancreas of the guinea-ping. Cells exhib-iting 54-HT immunoreactivity were found in the center and the periphery of the pancreatic islets. The5-HT immunoreactive granules were evenly distributed throughout the cytoplasm, but was faint inthe nuclei. Cells of this type were also distributed in the exocrine pancreas. Grouped or single or single cellsoccurred intercalatedly between acinar cells, as well as in the epithlium of ducts and in the connectivetissues near the ducts or acini.展开更多
BACKGROUND Mucinous adenocarcinomas of the buttock are rare and have an uncertain etiology and natural course.They are usually related to chronic anal fistulas,hidradenitis suppurativa,or Crohn's disease.Here,we r...BACKGROUND Mucinous adenocarcinomas of the buttock are rare and have an uncertain etiology and natural course.They are usually related to chronic anal fistulas,hidradenitis suppurativa,or Crohn's disease.Here,we report a case of mucinous adenocarcinoma associated with hidradenitis and contradictory immunochemistry results.CASE SUMMARY A 62-year-old man complained of recurrent abscesses of the buttock for 3 years.He had several scars and nodules in bilateral buttocks,with purulent discharge.The skin lesions did not appear to originate from the anus.The patient was diagnosed with recurrent abscesses due to hidradenitis suppurativa at the first visit.He showed purulent and subsequent mucin discharge in the first operation and was diagnosed with mucinous adenocarcinoma.Several examinations were performed to determine disease origin and staging.There were no significant findings or evidence of anal fistulas.Hence,he underwent wide local excision and V-Y advancement flap in the second operation.The final diagnosis was mucinous adenocarcinoma without any evidence of anal fistulas.Additional immunochemistry test results were negative for cytokeratin(CK)7 and positive for CK20 and CDX2,with a colorectal origin.A pathologist suggested that the disease originated from a chronic anal fistula.The patient has remained free of recurrence for 24 mo.CONCLUSION Although the patient with mucinous adenocarcinoma showed an atypical course,immunochemistry helped detect the disease origin.展开更多
Background: Determination of cyclosporine A (CsA) and tacrolimus (Tac) in dried blood spots (DBS) could enable drug monitoring in transplanted patients without the necessity of having to take venous blood samples. The...Background: Determination of cyclosporine A (CsA) and tacrolimus (Tac) in dried blood spots (DBS) could enable drug monitoring in transplanted patients without the necessity of having to take venous blood samples. Therefore, we have developed a method for quantitative determination of calcineurin inhibitors (CNI) by liquid-chromatography-tandem mass spectrometry (LCMS). Methods: In a study with 68 kidney transplant recipients (KTR, 34 CsA, 34 Tac), we tested the clinical application of LCMS monitoring in DBS in comparison to LCMS in whole blood. Results: The measuring range is proven for 27.33 to 1345 ng/ml for CsA and for 1.63 to 39.7 ng/ml for Tac. The requirements for clinical chemical analyses for precision and accuracy are complied with. Stability is documented for a period of 14 days. The study showed the following deviations from LCMS in whole blood for determination of CsA and Tac in DBS after introducing a correction factor by the haematocrit (Hct) value (CsA trough level: mean = 4.7%, ±1.96 standard deviation (SD) -52.1% to 61.4%, N = 96;CsA peak level: mean = 7.3%, ±1.96 SD -39.7% to 54.4%, N = 95;Tac trough level: mean = -0.5%, ±1.96 SD -76.4% to 75.3%, N = 88;Tac peak level: mean = 3.9%, ±1.96 SD -80.1% to 88.7%, N = 92). Conclusions: Our data show comparable results with the reference method by means of LCMS in whole blood. Therefore, DBS of KTR for determination of CNI levels could be transported on filter cards by mail to the respective laboratory resistant to breakage and the hazard of infection.展开更多
We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a mo...We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.展开更多
Ribulose-1, 5-bisphosphate carboxylase/oxygenase (EC 4, 1, 1, 39, abbreviation: RuBPcase)is a bifunction enzyme. Because of its properties in genetic function and structure, RuBPcase has been intensively studied. In t...Ribulose-1, 5-bisphosphate carboxylase/oxygenase (EC 4, 1, 1, 39, abbreviation: RuBPcase)is a bifunction enzyme. Because of its properties in genetic function and structure, RuBPcase has been intensively studied. In the absence of certain effective genetic markers for identifying hybrids, the analysis of the isoelectrofocusing pattern of RuBPcase has become a standard procedure to verify the nature of the hybrid plants.展开更多
基金Funded by the Natural Science Research Program of Jiangsu Education Department(No.13KJB310014)Natural Science Research Program of Jiangsu Province(No.BK20140429)
文摘The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothermal titration calorimetry(ITC) measurement. The results showed that there was competitive adsorption between heparin and fibronectin, and the preadsorption of fibronectin could prevent subsequent heparin adsorption to some extent, and the adsorbed Hep/Fn complex on the surface was in a rigid form. The bioactivity of heparin and fibronectin could be affected by the bulk concentration of each, and both heparin and fibronectin in Hep/Fn complex formed under p H 4 condition displayed larger bioactivity than that formed under p H 7 condition. Moreover, the fibronectin showed more exposed cell-binding sites at the p H value lower than physiological condition. The results of ITC further suggested that the interaction between heparin and fibronectin under p H 4 was stronger than under p H 7, and the complex was also more stable. The study brings forth the detailed interaction between heparin and fibronectin, which will be helpful for better understanding the interaction mechanism of the two biomolecules. The results may be potentially useful for the development of new generation of cardiovascular biomaterials.
基金the Asan Institute for Life Sciences,Asan Medical Center,Seoul,South Korea,No.2020IP0039.
文摘BACKGROUND The microsatellite instability(MSI)test and immunohistochemistry(IHC)are widely used to screen DNA mismatch repair(MMR)deficiency in sporadic colorectal cancer(CRC).For IHC,a two-antibody panel of MLH1 and MSH2 or four-antibody panel of MLH1,MSH2,PMS2,and MSH6 are used.In general,MSI is known as a more accurate screening test than IHC.AIM To compare two-and four-antibody panels of IHC in terms of accuracy and cost benefit on the basis of MSI testing for detecting MMR deficiency.METHODS We retrospectively analyzed patients with CRC who underwent curative surgery between 2015 and 2017 at a tertiary referral center.Both IHC with four antibodies and MSI tests were routinely performed.The sensitivity and specificity of a fourand two types of two-antibody panels(PMS2/MSH6 and MLH1/MSH2)were compared on the basis of MSI testing for detecting MMR deficiency.RESULTS High-frequency MSI was found in 5.5%(n=193)of the patients(n=3486).The sensitivities of the four-and two types of two-antibody panels were 97.4%,92.2%,and 87.6%,respectively.The specificities of the three types of panels did not differ significantly(99.6%for the four-antibody and PMS2/MSH6 panels,99.7%for the MLH1/MSH2 panel).Based on Cohen's kappa statistic(κ),four-and twoantibody panels were in almost perfect agreement with the MSI test(κ>0.9).The costs of the MSI test and the four-and two-antibody panels of IHC were approximately$200,$160,and$80,respectively.CONCLUSION Considering the cost of the four-antibody panel IHC compared to that of the twoantibody panel IHC,a two-antibody panel of PMS2/MSH6 might be the best choice in terms of balancing cost-effectiveness and accuracy.
文摘Immunocytochemical localization of 5-hydroxytryptamine (5-HT) containing cells of the guinea-pig pancreas was studied by means of PAP-glucose oxidase-DAB-nickel (PAP-GDN) and PAPstaining methods on paraffin sections. In this study we demonstrated the presence of 5-HTimmunoreactive cells in the endocrine (islets) and exocrine pancreas of the guinea-ping. Cells exhib-iting 54-HT immunoreactivity were found in the center and the periphery of the pancreatic islets. The5-HT immunoreactive granules were evenly distributed throughout the cytoplasm, but was faint inthe nuclei. Cells of this type were also distributed in the exocrine pancreas. Grouped or single or single cellsoccurred intercalatedly between acinar cells, as well as in the epithlium of ducts and in the connectivetissues near the ducts or acini.
文摘BACKGROUND Mucinous adenocarcinomas of the buttock are rare and have an uncertain etiology and natural course.They are usually related to chronic anal fistulas,hidradenitis suppurativa,or Crohn's disease.Here,we report a case of mucinous adenocarcinoma associated with hidradenitis and contradictory immunochemistry results.CASE SUMMARY A 62-year-old man complained of recurrent abscesses of the buttock for 3 years.He had several scars and nodules in bilateral buttocks,with purulent discharge.The skin lesions did not appear to originate from the anus.The patient was diagnosed with recurrent abscesses due to hidradenitis suppurativa at the first visit.He showed purulent and subsequent mucin discharge in the first operation and was diagnosed with mucinous adenocarcinoma.Several examinations were performed to determine disease origin and staging.There were no significant findings or evidence of anal fistulas.Hence,he underwent wide local excision and V-Y advancement flap in the second operation.The final diagnosis was mucinous adenocarcinoma without any evidence of anal fistulas.Additional immunochemistry test results were negative for cytokeratin(CK)7 and positive for CK20 and CDX2,with a colorectal origin.A pathologist suggested that the disease originated from a chronic anal fistula.The patient has remained free of recurrence for 24 mo.CONCLUSION Although the patient with mucinous adenocarcinoma showed an atypical course,immunochemistry helped detect the disease origin.
文摘Background: Determination of cyclosporine A (CsA) and tacrolimus (Tac) in dried blood spots (DBS) could enable drug monitoring in transplanted patients without the necessity of having to take venous blood samples. Therefore, we have developed a method for quantitative determination of calcineurin inhibitors (CNI) by liquid-chromatography-tandem mass spectrometry (LCMS). Methods: In a study with 68 kidney transplant recipients (KTR, 34 CsA, 34 Tac), we tested the clinical application of LCMS monitoring in DBS in comparison to LCMS in whole blood. Results: The measuring range is proven for 27.33 to 1345 ng/ml for CsA and for 1.63 to 39.7 ng/ml for Tac. The requirements for clinical chemical analyses for precision and accuracy are complied with. Stability is documented for a period of 14 days. The study showed the following deviations from LCMS in whole blood for determination of CsA and Tac in DBS after introducing a correction factor by the haematocrit (Hct) value (CsA trough level: mean = 4.7%, ±1.96 standard deviation (SD) -52.1% to 61.4%, N = 96;CsA peak level: mean = 7.3%, ±1.96 SD -39.7% to 54.4%, N = 95;Tac trough level: mean = -0.5%, ±1.96 SD -76.4% to 75.3%, N = 88;Tac peak level: mean = 3.9%, ±1.96 SD -80.1% to 88.7%, N = 92). Conclusions: Our data show comparable results with the reference method by means of LCMS in whole blood. Therefore, DBS of KTR for determination of CNI levels could be transported on filter cards by mail to the respective laboratory resistant to breakage and the hazard of infection.
文摘We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.
文摘Ribulose-1, 5-bisphosphate carboxylase/oxygenase (EC 4, 1, 1, 39, abbreviation: RuBPcase)is a bifunction enzyme. Because of its properties in genetic function and structure, RuBPcase has been intensively studied. In the absence of certain effective genetic markers for identifying hybrids, the analysis of the isoelectrofocusing pattern of RuBPcase has become a standard procedure to verify the nature of the hybrid plants.
