Deep learning-based recognition of stained tongue coating images

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.

Tectona grandis (Teak Tree) Young Leaf Extract as a Histological Stain

Stains are applied to impart contrast to the tissue and identify particular features of interest. However, the use of synthetic dyes as staining reagents has been associated with significant human health challenges and pollution of the ecosystem. These developments have necessitated a shift towards using natural dyes that are eco-friendlier and readily available. We investigated the staining reaction patterns of teak tree leaves (Tectona grandis) dye extracts and explored their suitability as a cytoplasmic stain in micromorphological assessments. Dye extracts were prepared using acetone, methanol, and ethanol as solvents from air-dried (under shade) teak tree young leaves. The dye extracts were applied as a counterstain and evaluated against eosin in formalin-fixed paraffin-embedded (FFPE) bovine tissue sections at varying concentrations and different staining times. Teak tree leaves (Tectona grandis) dye extracts produced relatively varying staining intensities of reddish-brown cytoplasmic coloration when used on bovine tissue at different concentrations and staining times comparable to eosin and with blue-purple hematoxylin nuclear stain. The present study showed that Tectona grandis leaf dye extracts provide an excellent cytoplasmic staining pattern and can be used as an alternative counterstain in routine H&E staining techniques.

The superiority and feasibility of 2,3,5-triphenyltetrazolium chloride-stained brain tissues for molecular biology experiments based on microglial properties

Background:TTC(2,3,5-triphenyltetrazolium chloride)staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model.Given that microglia exhibit different morphologies in different regions after ischemic stroke,we demonstrate the superiority and necessity of using TTC-stained brain tissue to analyze the expression of various proteins or genes in different regions based on microglia character.Methods:We compared brain tissue(left for 10 min on ice)from the improved TTC staining method with penumbra from the traditional sampling method.We identified the feasibility and necessity of the improved staining method using real time(RT)-PCR,Western blot,and immunofluorescence analysis.Results:There was no protein and RNA degradation in the TTC-stained brain tissue group.However,the TREM2 specifically expressed on the microglia showed a significant difference between two groups in the penumbra region.Conclusions:TTC-stained brain tissue can be used for molecular biology experiments without any restrictions.In addition,TTC-stained brain tissue shows greater superiority due to its precise positioning.

Can the Prediction of Intrauterine Insemination Results by Used Aniline Blue Stain (ABS) and Sperm Chromatin Dispersion (SCD) Levels?

Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine insemination before starting Intrauterine Insemination (IUI) method. Materials and Methods: In this prospective study, 111 couples who underwent Intrauterine Insemination (IUI) in unexplained infertility patients were admitted to Al-Farah IVF and assisted reproductive center in Baghdad, Iraq between November 2020 and February 2021 were evaluated. Semen fluid analysis was performed based on (WHO 4th) guiding rules. In addition, Sperm Chromatin Dispersion (halo test) and sperm maturation were performed with Aniline Blue Stain (ABS). Results: Sperm Chromatin Dispersion (SCD) groups were compared in terms of pregnancy outcome;the positive pregnancy rate was found to be above in the normal SCD groups (p = 0.0005). In addition, Aniline Blue Stain (ABS) groups were compared in the terms of pregnancy outcome;the positive pregnancy rate was found to be higher in the normal ABS group (p = 0.017). Conclusion: Our study showed that the use of DNA fragmentation (SCD) and sperm maturation tests (ABS) together with routine semen analysis in intrauterine insemination cases will make a significant contribution to the prediction of Intrauterine Insemination (IUI) increased results. So, these results indicate a defect in the effect of DNA fragmentation on the outcome of intrauterine insemination.

Advances in photodynamic therapy for port-wine stain and our experience

Port-wine stain(PWS)is a congenital capillary malformation that occurs in 0.3%–0.5%of newborns.The pulsed dye laser is the current gold standard treatment for PWS;however,its efficacy is poor.Photosensitizer photodynamic therapy(PDT)is considered a promising treatment for PWS.Here we provide a comprehensive overview of PDT.

A revisit to staining reagents for neuronal tissues

In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(and often naturally occurring)staining substances.Incidentally,the active ingredients of most of them were small molecules.With the advent of time,the knowledge of chemistry helped identify compounds and conditions for staining.The staining reagents were even found to enhance the visibility of the organelles.Silver impregnation identification of Golgi bodies was discovered in owl optic nerve.Staining reagents since the late 1800s were widely used across all disciplines and for nerve tissue and became a key contributor to advancement in nerve-related research.The use of these reagents provided insight into the organization of the neuronal tissues and helped distinguish nerve degeneration from regeneration.The neuronal staining reagents have played a fundamental role in the clinical research facilitating the identification of biological mechanisms underlying eye and neuropsychiatric diseases.We found a lack of systematic description of all staining reagents,whether they had been used historically or currently used.There is a lack of readily available information for optimal staining of different neuronal tissues for a given purpose.We present here a grouping of the reagents based on their target location:(I)the central nervous system(CNS),(II)the peripheral nervous system(PNS),or(III)both.The biochemical reactions of most of the staining reagents is based on acidic or basic pH and specific reaction partners such as organelle or biomolecules that exists within the given tissue type.We present here a summary of the chemical composition,optimal staining condition,use for given neuronal tissue and,where possible,historic usage.Several biomolecules such as lipids and metabolites lack specific antibodies.Despite being non-specific the reagents enhance contrast and provide corroboration about the microenvironment.In future,these reagents in combination with emerging techniques such as imaging mass spectrometry and kinetic histochemistry will validate or expand our understanding of localization of molecules within tissues or cells that are important for ophthalmology and vision science.

Screening of some indigenous herbal dyes for use in plant histological staining

The efficacies of some indigenous herbal dyes for use in staining plant materials were examined to obtain non-toxic, eco-friendly and cheap stains for use in plant histology. Dye extracts from Bixa orellana, Curcuma domestica, Lonchocarpus cyanescens and Pterocarpus osun were used to stain wood sections using the existing standard staining procedures with little modification. All the extracts had affinity for the fibre and vessel elements except the extract from L. cyanescens. The extracts from C. domestica and B. orellana had higher selectivity than those ofP. osun for fibre. From the results of the absorbance curves, each of the dye extracts from all speciese had minimum of two peaks, indicating that they had two or more colour imparting chromophores except dye extract from C. domestica. All the dye extracts were acidic with pH range of 3.77 to 6.77. Therefore, this study shows that dye extracts from B. orellana, C. domestica and P. osun could be solitarily or in combination with artificial dyes for plant histological staining.

Comparison of Sensitivity and Specificity of ZN and Fluorescent Stain Microscopy with Culture as Gold Standard

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.

CHANGES OF VISUAL FINDINGS, ELECTRIC FEATURES AND STAINING OF AURICLES IN MALIGNANT TUMOR PATIENTS

In order to find a simple and conve-nient method of diagnosis and to probe intothe value of auricular diagnosis in malignanttumors so as to substantiate and

Comparative Investigation of Alternative Negative Staining Reagents in Bacterial Morphological Study

Negative staining is an effective method that can be used for electron microscopic study to observe fine structural morphology without destruction of bacterial structure. Although uranium acetate is used worldwide as a general dyeing solution, it is extremely difficult to use it by a new purchase at a research institution because it falls under the nuclear regulation substance in Japan. Therefore, we examined alternative reagents for negative staining that could replace uranium acetate through bacterial observation with an electron microscope. Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pyogenes were examined by four stain reagents (phosphotungstic acid (PTA), EMstainer, TI blue, and uranium acetate). Pre cultured bacteria were stained with each stain reagents on a copper grid, washed with PBS, and observed with a transmission electron microscope. In the comparison between bacterial structures, the cell wall structure and bacterial flagella could be observed well in the order of PTA, EMstainer, and uranium acetate. With TI blue staining, flagella could be observed very poorly. In comparison between bacteria, gram negative bacteria such as Escherichia coli and Pseudomonas aeruginosa, could be observed well as compared with gram positive cocci such as Staphylococcus aureus and Streptococcus pyogenes. The uranium acetate looked very coarse in background particles. Since crystals tend to precipitate, TI blue also required filtering, and electron beams were absorbed by the agglomerated crystals, and the frequency of electronic burning occurred high frequency. In this study, there was clear difference in the observation conditions depending on the type of bacteria and the kind of the staining reagents. Especially, it was confirmed that good negative staining features of Pseudomonas aeruginosa by electron microscope were obtained by PTA and EMstainer staining. These alternative reagents are considered to be a candidate for a negative staining.

A RAPID STAINING METHOD SHOWING AgNOR AND DNA IN PATHOLOGICAL DIAGNOSIS

In order to satisfy the need of rapid clinical diagnosis, a series of experimental studies for AgNOR (Argyrophilic NORassociated proteins) and DNA (Dereyribonucleic acid) staining was compared, from which a stable staining solution was selected to show mucin residence. Using the referred tables, with reference to the corresponding relation of the staining solution with temperature and time, reliable staining results for AgNOR and DNA could be obtained.These two modified staining methods are simple, reliable and easy to operate, able to provide good contrast for pathological diagnosis.

Experimental Study on the Effectiveness and Safety of Gardenia Blue Pigment on Anterior Capsule Staining

Objective: To explore the effect of gardenia blue pigment on the anterior capsule staining and its safety performance. Methods: In this study, New Zealand white rabbits were used as the research object to test the effect of gardenia blue pigment on the anterior capsule staining and anterior segment toxicity. In this study, gardenia blue stains of different concentrations (5%, 4%, 3%, 2%, 1%, 0.5%) were prepared to stain the anterior capsule of rabbit crystals, and the effect of the stain was observed to determine the optimal concentration of the stain. Twenty-seven healthy New Zealand white rabbits without eye disease were randomly divided into three groups: gardenia blue staining group;bio-blue positive drug group;physiological saline group. The intraocular pressure, anterior chamber inflammation, corneal endothelial injury, and anterior chamber angle pathological changes were measured in 1D, 7D, and 14D. Results: The results of this study showed that the effect of gardenia blue pigment on the anterior capsule of the lens was the best when the concentration of blue pigment was 2%. There was no change in intraocular pressure in the anterior chamber for a short period of time. There is no damage to the corneal endothelium. Conclusion: The results of this study show that the effect of gardenia blue pigment on the anterior capsule is best when the concentration of blue pigment is 2%, and the stain has no obvious toxic and side effects on the anterior segment of the eye.

Stain Materials’Role in Biological Research:A Tool in Heath Care

Stains and staining methods significantly assist in diagnoses in medical research and health care.Certain color of a dye can identify the location of a tumor within a specimen.Applying histochemical-staining enabled morphological identification of fibrin in the lymphoid tissue during cancer progression.Staining methods in combination with the LSFM(light-sheet fluorescence microscopy)allowed tracing the drug penetration,development and spread of tumors.The Curcumin dye is in use for labelling and imaging of Aβplaques in post-mortem brain tissue.Immunofluorescent staining methods are employed in detection of some important proteins in early diagnostic changes relevant to heart damage.The methods are developed in medical research to include stem cells and tissue engineering,cell culturesproperties and capabilities,connective tissues and extracellular matrix,nervous system,musculoskeletal system;respiratory system,liver and gastrointestinal tract,and male and female reproductive systems.

On the Multiple Narrative Strategies in The Human Stain

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.

Comparative Study of Z N Staining vs. Flurochrome Staining and Impact of Sample Processing on Diagnosis of Tuberculosis from Various Clinical Samples

Background: Tuberculosis is a highly infectious disease and India has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. Although its sensitivity is low in comparison to culture and molecular methods, its sensitivity can still be improved by using fluorescence staining method and processing of samples by homogenization and concentration method. Material and methods: Samples were collected from all newly registered suspected cases of tuberculosis in tertiary care hospital from outward and indoor department during a period of one year. Smears were prepared for Ziehl Neelsen stain and fluorescence stain both before and after homogenization and concentration procedure by 4% NAOH-2.9% sodium citrate method and results of them were interpreted according to RNTCP criteria for grading of sputum samples. All the samples were cultured in liquid culture MGIT system (Mycobacterial Growth Indicator Tube) and results of microscopy were compared with liquid culture taken as gold standard. Data were analyzed by using SPSS software version 16. Result: 350 samples were collected during study period. Out of 350 samples, 48 samples were positive for M. tuberculosis by MGIT system. In comparison with MGIT system, sensitivity of Z N stain for detection of acid fast bacilli was 77% before decontamination procedure, which was increased up to 85.42% after decontamination and concentration process. Sensitivity of fluroscence stain was 85.42% before processing, which was increased up to 91.67% after processing of samples. Conclusion: Sensitivity of smear microscopy can be enhanced by use of fluroscence microscopy and concentration method.

RAL-STAINER自动染色仪对抗酸染色的性能评估

目的 评估RAL-STAINER自动染色仪对抗酸染色的性能。方法 收集标本,分别进行手工抗酸染色和仪器抗酸染色,评价自动染色仪的染色效果的合格率与手工染色的染色结果的一致率、仪器染色的重复性、与室间质评预期结果的符合率、交叉污染和工作性能等。结果 RAL-STAINER自动染色仪对各类标本抗酸染色背景基本无杂质,细菌和细胞色泽鲜明、结构清楚,染色效果的合格率为100%。自动染色仪与手工抗酸染色一致率为100%。仪器染色的重复性为100%。使用自动染色仪,两次室间质评结果均100%符合预期。自动染色仪批量染色时,无交叉污染。自动染色仪操作简便、通量高、染色效果稳定、生物安全性好。结论RAL-STAINER自动染色仪对抗酸染色的性能较好。

Selection of oocytes for in vitro maturation by brilliant cresyl blue staining： a study using the mouse model

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.

A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy

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.

Application of an indirect immunofluorescent staining method for detection of Salmonella enteritidis in paraffin slices and antigen location in infected duck tissues

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.

Helicobacter pylori infection in the pharynx of patients with chronic pharyngitis detected with TDI-FP and modified Giemsa stain

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.