Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,vi...Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,visualizing conjugation in ciliates is a challenge due to the complexity and dynamics of the process,while traditional staining methods are often insufficient for the research.This study introduces a new method for visualizing developmental progression in the nuclei during conjugation using Hoechst33342 staining.It describes how to proceed from cell culture,conjugation induction and synchronization,staining preparation,and observation to statistical analysis.The combination of fluorescent staining with the‘volume-fixing'technique eliminates the fixation and dehydration steps,thus reducing the overall operation time to just 20 minutes.This method offers several advantages over traditional staining techniques for studying the nuclei during conjugation.It improves image quality and workflow efficiency and enables real-time observation of live cell states.Potential solutions to challenges that may arise during experimental procedures are introduced and references and guidelines for cytological research are provided in this paper.展开更多
Objective To build a dataset encompassing a large number of stained tongue coating images and process it using deep learning to automatically recognize stained tongue coating images.Methods A total of 1001 images of s...Objective To build a dataset encompassing a large number of stained tongue coating images and process it using deep learning to automatically recognize stained tongue coating images.Methods A total of 1001 images of stained tongue coating from healthy students at Hunan University of Chinese Medicine and 1007 images of pathological(non-stained)tongue coat-ing from hospitalized patients at The First Hospital of Hunan University of Chinese Medicine withlungcancer;diabetes;andhypertensionwerecollected.Thetongueimageswererandomi-zed into the training;validation;and testing datasets in a 7:2:1 ratio.A deep learning model was constructed using the ResNet50 for recognizing stained tongue coating in the training and validation datasets.The training period was 90 epochs.The model’s performance was evaluated by its accuracy;loss curve;recall;F1 score;confusion matrix;receiver operating characteristic(ROC)curve;and precision-recall(PR)curve in the tasks of predicting stained tongue coating images in the testing dataset.The accuracy of the deep learning model was compared with that of attending physicians of traditional Chinese medicine(TCM).Results The training results showed that after 90 epochs;the model presented an excellent classification performance.The loss curve and accuracy were stable;showing no signs of overfitting.The model achieved an accuracy;recall;and F1 score of 92%;91%;and 92%;re-spectively.The confusion matrix revealed an accuracy of 92%for the model and 69%for TCM practitioners.The areas under the ROC and PR curves were 0.97 and 0.95;respectively.Conclusion The deep learning model constructed using ResNet50 can effectively recognize stained coating images with greater accuracy than visual inspection of TCM practitioners.This model has the potential to assist doctors in identifying false tongue coating and prevent-ing misdiagnosis.展开更多
One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies ...One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies that Philip R oth employs in the novel.T his paper focuses mainly on the elaboration of such narrative strategies as mimesis and diegesis,complicated narrative time,shifting focalizations,and narration or non-narrative comments,so as to point out that these narrative strategies add colors to its( postmodern) artistic features,becoming an integral component of this novel.展开更多
AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in O...AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. ' RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group. CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.展开更多
In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4＇, 6-diamidino-2-phenylindole （DAPI） mult...In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4＇, 6-diamidino-2-phenylindole （DAPI） multiple bands like （J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.展开更多
We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobrom...We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobromae, carbon and oxygen elements and the ratio of oxygen to carbon decreased. Considering atomic binding,C_1 and C_4 contents increased, while C_2 and C_3 contents decreased, and the ratio of O_2 to O_1 decreased sharply. The relative contents of lignin, cellulose and polysaccharides increased and new substances with low O_2/O_1 ratio occurred. FTIR analysis showed that the absorption peaks of O–H at 3346 cm^(-1), aliphatic C–H at 2921, 2853 and1464 cm^(-1), and C=O at 1723 cm^(-1), were characteristic peaks of fungal melanin intensified, indicating that cane discoloration was primarily caused by fungal melanin. The absorption peaks characterizing cellulose and lignin like polysaccharides at 800 cm^(-1), C–H at 1374 cm^(-1), C–O at1058 and 1038 cm^(-1), phenolic hydroxyl at 1245 cm^(-1),aromatic ether bonds at 1270 cm^(-1), carbon skeleton at1608 cm^(-1) and benzene ring at 1500 cm^(-1) were enhanced since the fungus mainly consumed the extractives in cane cell lumens and the main composition content increased relatively. Regardless of the discoloration caused by natural fungi or inoculated fungi, the discoloring feature and composition changes were identical except that the fungusinoculated cane had more melanin.展开更多
Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider uti...Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.展开更多
AIM： To detect whether there is Helicobacter pylori （Hpylori） colonization in the pharynx mucous membrane of healthy people and whether chronic pharyngitis is related to Hpylori infection. METHODS： Fifty cases of ...AIM： To detect whether there is Helicobacter pylori （Hpylori） colonization in the pharynx mucous membrane of healthy people and whether chronic pharyngitis is related to Hpylori infection. METHODS： Fifty cases of chronic pharyngitis refractory over three months were prospectively studied from March 2004 to August 2004 in the otolaryngology outpatient department of the Second Hospital of Xi＇an Jiaotong University. Template-directed dye-terminator incorporated with fluorescence polarization detection （TDI-FP） and modified Giemsa stain were used to examine pharynx mucous membrane tissue for H pylori colonization in the patients with chronic pharyngitis and the healthy people as a control group. RESULTS： In the control group, no people were detected to have Hpylori in the pharynx. In contrast, in 50 cases with chronic pharyngitis, 19 （38.0%） cases were H pylori positive with a TDI-FP assay and 4 （8%） cases were TDI-FP positive with Giemsa staining in the pharynx. Sixteen of the 50 pharyngitis cases had stomach ailment history, 11 cases （68.8%） of these 16 patients were determined to be H pylori positive in the pharynx with the TDI-FP assay. 2,2 test showed that this infection rate was remarkably higher （P= 0.0007） than that in the cases without stomach ailment history. Giemsa staining showed that 3 cases （18.8%） of the patients with stomach ailment history were infected with H pylori in the pharynx, which was remarkably higher （P = 0.042） than that in the patients without stomach ailment history （1 case, which was 2.9%）. CONCLUSION： H pylori may not be detected in the pharynx of healthy people. Chronic pharyngitis may be related to H pylori infection. The infection rate with Hpylori in the pharynx is higher in patients with stomach ailment histories than in patients without stomach ailment histories, suggesting that chronic pharyngitis may be related to stomach ailment history.展开更多
AIM： To detect Salmonella enteritidis （S. enteritidis） in paraffin slices and antigen location in infected duck tissues. METHODS： The rabbits were immunized with purified bacillus to obtain S. enteritidis-specific...AIM： To detect Salmonella enteritidis （S. enteritidis） in paraffin slices and antigen location in infected duck tissues. METHODS： The rabbits were immunized with purified bacillus to obtain S. enteritidis-specific antibody, which were then extracted by the caprylic-ammonium sulphate method, purified through High-Q columns. An indirect immuno-fluorescent staining method （IFA） was established to detect the S. enteritidis antigen in paraffin slices. Detected S. enteritidis in each organ tissue of ducklings experimentally infected with S. enteritidis. RESULTS： The gland of Garder, heart, kidney, spleen, liver, brain, ileum, jejunum, bursa of Fabricius from S. enteritidis experimentally infected ducklings were positive or strongly positive, and the S. enteritidis antigen mainly distributed in the infected cell cytoplasm.CONCLUSION： IFA is an intuitionist, sensitive and specific method in detecting S. enteritidis antigen in paraffin wax slices, and it is a good method in diagnosis and antigen location of S. enteritidis. We also conclude that the gland of Garder, heart, kidney, spleen, liver, ileum, jejunum are target organs in S. enteritidis infections of duck, and S. enteritidis is an intracellular parasitic bacterium.展开更多
Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and spu...Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and sputum concentration may also improve sensitivity of microscopy. Objective: To compare the sensitivity and specificity of microscopy for AFB using ZN and fluorescent stains in direct and concentrated specimen with culture as gold standard. Methods: Morning sputum specimen of patients, suspected of having pulmonary tuberculosis, over a period of 6 months was subjected to direct microscopy using fluorescent stain;the same slide was over-stained with ZN stain. Same sputum sample was concentrated by Petroff’s method and subjected to fluorescent microscopy followed by ZN microscopy and finally to culture for AFB. Results: Sensitivity of fluorescent stained concentrated sputum samples was maximum and of ZN stained unprocessed sputum samples was minimum. Specificity of three of the methods was equal at 0.96 but of ZN stained concentrated sputum smears was 0.97. Sensitivity of total fluorescent stains was 0.85 (Specificity 0.96) and sensitivity of total ZN stained smears was 0.80 (Specificity 0.96). Discussion: We used same smear for fluorescent and ZN stains, so smear related variability is decreased. Blinding for microscopy was practically complete. Conclusion: The sensitivity of sputum microscopy for AFB can be increased by concentrating the sputum and using fluorescent microscopy. The specificity remains high in all the methods.展开更多
基金supported by the National Natural Science Foundation of China(No.32270558)。
文摘Ciliates are eukaryotic unicellular organisms with complex morphology and developmental processes,including asexual and sexual processes.Conjugation is a form of sexual process that renews genetic materials.However,visualizing conjugation in ciliates is a challenge due to the complexity and dynamics of the process,while traditional staining methods are often insufficient for the research.This study introduces a new method for visualizing developmental progression in the nuclei during conjugation using Hoechst33342 staining.It describes how to proceed from cell culture,conjugation induction and synchronization,staining preparation,and observation to statistical analysis.The combination of fluorescent staining with the‘volume-fixing'technique eliminates the fixation and dehydration steps,thus reducing the overall operation time to just 20 minutes.This method offers several advantages over traditional staining techniques for studying the nuclei during conjugation.It improves image quality and workflow efficiency and enables real-time observation of live cell states.Potential solutions to challenges that may arise during experimental procedures are introduced and references and guidelines for cytological research are provided in this paper.
基金National Natural Science Foundation of China(82274411)Science and Technology Innovation Program of Hunan Province(2022RC1021)Leading Research Project of Hunan University of Chinese Medicine(2022XJJB002).
文摘Objective To build a dataset encompassing a large number of stained tongue coating images and process it using deep learning to automatically recognize stained tongue coating images.Methods A total of 1001 images of stained tongue coating from healthy students at Hunan University of Chinese Medicine and 1007 images of pathological(non-stained)tongue coat-ing from hospitalized patients at The First Hospital of Hunan University of Chinese Medicine withlungcancer;diabetes;andhypertensionwerecollected.Thetongueimageswererandomi-zed into the training;validation;and testing datasets in a 7:2:1 ratio.A deep learning model was constructed using the ResNet50 for recognizing stained tongue coating in the training and validation datasets.The training period was 90 epochs.The model’s performance was evaluated by its accuracy;loss curve;recall;F1 score;confusion matrix;receiver operating characteristic(ROC)curve;and precision-recall(PR)curve in the tasks of predicting stained tongue coating images in the testing dataset.The accuracy of the deep learning model was compared with that of attending physicians of traditional Chinese medicine(TCM).Results The training results showed that after 90 epochs;the model presented an excellent classification performance.The loss curve and accuracy were stable;showing no signs of overfitting.The model achieved an accuracy;recall;and F1 score of 92%;91%;and 92%;re-spectively.The confusion matrix revealed an accuracy of 92%for the model and 69%for TCM practitioners.The areas under the ROC and PR curves were 0.97 and 0.95;respectively.Conclusion The deep learning model constructed using ResNet50 can effectively recognize stained coating images with greater accuracy than visual inspection of TCM practitioners.This model has the potential to assist doctors in identifying false tongue coating and prevent-ing misdiagnosis.
文摘One of the significant features of The Human Stain is that it lays bare the reality of American society and censures the Anglo-Saxon or white discourse.What are equally important are the multiple narrative strategies that Philip R oth employs in the novel.T his paper focuses mainly on the elaboration of such narrative strategies as mimesis and diegesis,complicated narrative time,shifting focalizations,and narration or non-narrative comments,so as to point out that these narrative strategies add colors to its( postmodern) artistic features,becoming an integral component of this novel.
基金Supported by National Natural Science Foundation of China(No.30973899)
文摘AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. ' RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group. CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.
文摘In this study, mitotic metaphase chromosomes in mouse were identified by a new chromosome fluorescence banding technique combining DAPI staining with image analysis. Clear 4＇, 6-diamidino-2-phenylindole （DAPI） multiple bands like （J-hands could be produced in mouse. The Meta- Morph software was then used to generate linescans of pixel intensity for the banded chromosomes from short arm to long arm. These linescans were sufficient not only to identify each individual chromosome but also analyze the physical sites of bands in chromosome. Based on the results, the clear and accurate karyotype of mouse metaphase chromosomes was established. The technique is therefore considered to he a new method for cytological studies of mouse.
基金supported by the National Key Project of S&T Supporting Programs Funded by MOST of China during the 12th Five-year Plan(No.2012BAD23B0104)
文摘We explored the discoloration of rattan cane using X-ray photoelectron spectroscopy(XPS) and Fourier transform infrared spectroscopy(FTIR). XPS analysis showed that after the cane was stained by Lasiodiplodia theobromae, carbon and oxygen elements and the ratio of oxygen to carbon decreased. Considering atomic binding,C_1 and C_4 contents increased, while C_2 and C_3 contents decreased, and the ratio of O_2 to O_1 decreased sharply. The relative contents of lignin, cellulose and polysaccharides increased and new substances with low O_2/O_1 ratio occurred. FTIR analysis showed that the absorption peaks of O–H at 3346 cm^(-1), aliphatic C–H at 2921, 2853 and1464 cm^(-1), and C=O at 1723 cm^(-1), were characteristic peaks of fungal melanin intensified, indicating that cane discoloration was primarily caused by fungal melanin. The absorption peaks characterizing cellulose and lignin like polysaccharides at 800 cm^(-1), C–H at 1374 cm^(-1), C–O at1058 and 1038 cm^(-1), phenolic hydroxyl at 1245 cm^(-1),aromatic ether bonds at 1270 cm^(-1), carbon skeleton at1608 cm^(-1) and benzene ring at 1500 cm^(-1) were enhanced since the fungus mainly consumed the extractives in cane cell lumens and the main composition content increased relatively. Regardless of the discoloration caused by natural fungi or inoculated fungi, the discoloring feature and composition changes were identical except that the fungusinoculated cane had more melanin.
基金Acknowledgments This study was supported by grants from the China National Natural Science Foundation （Nos. 30430530 and 30571337） and from the Momentous Research Project of the China Ministry of Science and Technology （No. 2006CB944003）.
文摘Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.
基金Supported by a grant from the Bureau of Health in Shaanxi Province,No.2002 02D24 and grants No.NSFC30440080No.NIDCD R21 DC005846
文摘AIM： To detect whether there is Helicobacter pylori （Hpylori） colonization in the pharynx mucous membrane of healthy people and whether chronic pharyngitis is related to Hpylori infection. METHODS： Fifty cases of chronic pharyngitis refractory over three months were prospectively studied from March 2004 to August 2004 in the otolaryngology outpatient department of the Second Hospital of Xi＇an Jiaotong University. Template-directed dye-terminator incorporated with fluorescence polarization detection （TDI-FP） and modified Giemsa stain were used to examine pharynx mucous membrane tissue for H pylori colonization in the patients with chronic pharyngitis and the healthy people as a control group. RESULTS： In the control group, no people were detected to have Hpylori in the pharynx. In contrast, in 50 cases with chronic pharyngitis, 19 （38.0%） cases were H pylori positive with a TDI-FP assay and 4 （8%） cases were TDI-FP positive with Giemsa staining in the pharynx. Sixteen of the 50 pharyngitis cases had stomach ailment history, 11 cases （68.8%） of these 16 patients were determined to be H pylori positive in the pharynx with the TDI-FP assay. 2,2 test showed that this infection rate was remarkably higher （P= 0.0007） than that in the cases without stomach ailment history. Giemsa staining showed that 3 cases （18.8%） of the patients with stomach ailment history were infected with H pylori in the pharynx, which was remarkably higher （P = 0.042） than that in the patients without stomach ailment history （1 case, which was 2.9%）. CONCLUSION： H pylori may not be detected in the pharynx of healthy people. Chronic pharyngitis may be related to H pylori infection. The infection rate with Hpylori in the pharynx is higher in patients with stomach ailment histories than in patients without stomach ailment histories, suggesting that chronic pharyngitis may be related to stomach ailment history.
基金the National Key Technology R&D Program of China, No. 2004BA 901A 03National Scientific and Sechnical Support Program, No. 2007Z06-017+3 种基金The Cultvation Fund of the Key Scientific and Technical Innovation Project & Ministry of Education of China, No. 706050Program for New Century Excellent Talents in University, No. NCET-04-0906/NCET-06-0818Sichuan Province Basic Research Program, No. 04JY0290061/07JY029-017Program for Key Disciplines Construction of Sichuan Province No. SZD0418
文摘AIM： To detect Salmonella enteritidis （S. enteritidis） in paraffin slices and antigen location in infected duck tissues. METHODS： The rabbits were immunized with purified bacillus to obtain S. enteritidis-specific antibody, which were then extracted by the caprylic-ammonium sulphate method, purified through High-Q columns. An indirect immuno-fluorescent staining method （IFA） was established to detect the S. enteritidis antigen in paraffin slices. Detected S. enteritidis in each organ tissue of ducklings experimentally infected with S. enteritidis. RESULTS： The gland of Garder, heart, kidney, spleen, liver, brain, ileum, jejunum, bursa of Fabricius from S. enteritidis experimentally infected ducklings were positive or strongly positive, and the S. enteritidis antigen mainly distributed in the infected cell cytoplasm.CONCLUSION： IFA is an intuitionist, sensitive and specific method in detecting S. enteritidis antigen in paraffin wax slices, and it is a good method in diagnosis and antigen location of S. enteritidis. We also conclude that the gland of Garder, heart, kidney, spleen, liver, ileum, jejunum are target organs in S. enteritidis infections of duck, and S. enteritidis is an intracellular parasitic bacterium.
文摘Introduction: Reports indicate that fluorescent staining of smears increases sensitivity of direct microscopy;so ZN staining is being replaced with fluorescent microscopy in RNTCP in India. Chemical processing and sputum concentration may also improve sensitivity of microscopy. Objective: To compare the sensitivity and specificity of microscopy for AFB using ZN and fluorescent stains in direct and concentrated specimen with culture as gold standard. Methods: Morning sputum specimen of patients, suspected of having pulmonary tuberculosis, over a period of 6 months was subjected to direct microscopy using fluorescent stain;the same slide was over-stained with ZN stain. Same sputum sample was concentrated by Petroff’s method and subjected to fluorescent microscopy followed by ZN microscopy and finally to culture for AFB. Results: Sensitivity of fluorescent stained concentrated sputum samples was maximum and of ZN stained unprocessed sputum samples was minimum. Specificity of three of the methods was equal at 0.96 but of ZN stained concentrated sputum smears was 0.97. Sensitivity of total fluorescent stains was 0.85 (Specificity 0.96) and sensitivity of total ZN stained smears was 0.80 (Specificity 0.96). Discussion: We used same smear for fluorescent and ZN stains, so smear related variability is decreased. Blinding for microscopy was practically complete. Conclusion: The sensitivity of sputum microscopy for AFB can be increased by concentrating the sputum and using fluorescent microscopy. The specificity remains high in all the methods.
