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.

Application of short-pulsed CO_(2) laser and long-pulsed Nd:YAG laser in the treatment of severe hypertrophic port-wine stains

Background:The treatment of port-wine stains(PWS)becomes extremely difficult due to age-related malformation of the vasculature.In this study,we used alternative methods to treat hypertrophic PWS.Methods:A short-pulsed CO_(2) laser was applied to ablate the hypertrophic vasculature of PWS.The ablation was ended when the wound was on the same plane as normal skin.The long-pulsed neodymium-doped yttriumaluminum-garnet(lpNd:YAG)laser was primarily applied to coagulate and subsequently liquefy the hypertrophic vasculature of the PWS.The therapeutic energy used in treating different lesions should be carefully regulated to significantly affect the treatment outcomes.Results:The two cases presented herein demonstrated substantial improvement in hypertrophic vasculature that was largely removed.The skin was resurfaced,although some scar formation and mild hypopigmentation occurred.Conclusion:We suggest the use of short-pulsed CO_(2) and lpNd:YAG lasers for treating certain cases of hypertrophic PWS.When using a short-pulsed CO_(2) laser,it is wise to judge the treatment endpoint and take appropriate precautions to avoid intraoperative bleeding.When using an lpNd:YAG laser,the therapeutic energy should be controlled according to the thickness of the lesion to reduce scar formation.

Angiogenesis and proliferation of endothelial cells in hypertrophic and nodular port-wine stain

Background:Port-wine stain(PWS)has been classified not as the hyperplasia of cells,but rather,as an expansion of malformed vessels.However,previous studies have reported upregulated expression of proangiogenic factors in PWS.Several studies have indicated that the pathology exhibits proliferation of numerous endothelial cells in hypertrophic/nodular PWS.This study aimed to determine the expression of vascular epithelial growth factor(VEGF),matrix metalloproteinase-9(MMP-9),angiopoietin-2(ANG-2),and basic fibroblast growth factor(bFGF)in hypertrophic PWS.Methods:Immunohistochemistry was used to analyze skin samples from 33 patients with hypertrophic PWS.Expression levels of VEGF,MMP-9,ANG-2,and bFGF in hypertrophic PWS were determined by multiplying the intensity by the percentage of immunoreactive cells.Immunoreactivity scores were classified as follows:negative(0),low(1),moderate(2,3,and 4),or high(6).Results:Based on pathological characteristics,hypertrophic PWS was divided into vascular malformation and pyogenic granuloma(PG)types.VEGF,MMP-9,ANG-2,and bFGF were significantly activated in the blood vessels of PG-type PWS samples compared with their counterparts in blood vessels of vascular malformation-type PWS samples and controls.PG-type hypertrophic PWS,which exhibited proliferation of endothelial cells,showed the strongest activation.Conclusion:The exuberant proliferation of endothelial cells in PG-type hypertrophic PWS may be associated with the regulation of proangiogenic factors during development.These proangiogenic factors that function in the angiogenesis and proliferation of endothelial cells may play an important role in the pathogenesis and progression of PWS.Furthermore,these factors may be dynamic and behave differently in various types of hypertrophic PWS.

Glaucoma in Patients with Eyes Close to Areas Affected by Port-wine Stain has Lateral and Gender Predilection

Background：The location of facial port-wine stain （PWS） may be helpful for predicting some associated anomalies;high glaucoma incidence is found in patients with eyes close to PWS-affected areas （V1,ophthalmic branch area of the trigeminal nerve）.This study aimed to investigate the characteristics of glaucoma in V1-affected PWS.Methods：A total of 569 patients with V1 area-affected PWS were reviewed in the study.The large series was based on the referral system between the Department of Plastic and Reconstructive Surgery and the Department of Ophthalmology.All patients were screened for glaucoma with assessments of intraocular pressure,cup-to-disc ratio,comeal diameter （only for infants）,and axial length.Results：Of the 569 patients,110 （19.3%） patients had glaucoma.Among the patients,18.1% （76/420） had early-onset glaucoma （under 4-year-old group）.In the 4 to 18-year-old age group,29.3% （29/99） of the patients had glaucoma.Compared with right lateral and bilateral PWS,left-sided PWS had a lower risk of glaucoma in this study （odds ratio =0.432 [95% confidence interval,0.264-0.706],P =0.01）.The under 4-year-old group showed a slight predominance of males （61.8%） in glaucoma.Conclusions：High glaucoma incidence was observed in patients with eyes close to PWS.More attention should be paid to glaucoma screening for right lateral and bilateral PWS patients.The predominance of males in Sturge-Weber syndrome （SWS） early-onset glaucoma patients might be due to the limitation of the case number;however,it might also provide us a new clue of potential relationship between SWS and PCG.

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.

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.

亚砷酸钠对人正常肝细胞脂质代谢及因子调控的作用

背景:肝脏作为砷毒作用的主要靶器官,成为砷毒性作用机制相关研究的焦点。目的:探讨亚砷酸钠对人正常肝细胞脂质代谢、细胞增殖、细胞凋亡及相关调控因子表达的影响。方法:MIHA正常人肝细胞株暴露于0,10,20,30μmol/L亚砷酸钠48 h,光学显微镜观察细胞形态变化,CCK-8法检测细胞活性,单试剂COD-PAP法、单试剂GPO-PAP法及酶微板法检测细胞上清总胆固醇、三酰甘油及总胆汁酸水平,油红O染色法检测细胞内脂质含量,Edu-488渗入法检测细胞增殖,PI染色法及Annexin V-FITC/PI双标记法结合流式细胞术分别检测细胞周期和细胞凋亡,实时荧光定量PCR和Western blot检测肝细胞核因子4α、胆固醇7α-羟化酶及法尼醇X受体的m RNA及蛋白表达水平。结果与结论:(1)与对照组(0μmol/L亚砷酸钠)相比,随着亚砷酸钠浓度的增加:MIHA细胞活性逐渐降低;细胞上清中总胆固醇、三酰甘油水平逐渐增高,总胆汁酸水平逐渐降低;细胞内脂质含量逐渐增多;细胞停滞于S期及G_2/M期细胞比例逐渐增多;细胞凋亡率逐渐升高;肝细胞核因子4α mRNA表达水平未见明显变化,胆固醇7α-羟化酶与法尼醇X受体mRNA表达水平下降;肝细胞核因子4α、胆固醇7α-羟化酶、法尼醇X受体蛋白表达水平逐渐下降;(2)亚砷酸钠具有细胞毒性,显著降低MIHA细胞活性,诱导细胞脂肪变性,阻滞细胞增殖,诱发细胞凋亡等损伤;亚砷酸钠可下调肝细胞核因子4α蛋白表达以及胆固醇7α-羟化酶、法尼醇X受体的转录和蛋白表达,进一步引起肝细胞的脂质代谢紊乱。

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。

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.

Mouse Karyotype Obtained by Combining DAPI Staining with Image Analysis

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.

XPS and FTIR studies of fungus-stained Daemonorops margaritae

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.

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.

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.

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.

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.

Comparison ofβ-Amyloid Plaque Labeling Methods:Antibody Staining,Gallyas Silver Staining,and Thioflavin-S Staining

Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods.

Application of Prussian blue staining in the diagnosis of ocular siderosis

AIM:To explore the value of Prussian blue staining in the diagnosis of ocular siderosis.METHODS:Between January 2012 and January 2013,the Prussian blue stain used in anterior lens capsule and vitreous liquid after centrifugation from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis. At the same time, give a negative control.RESULTS:Anterior lens capsule membrane and liquid of vitreous cavity from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis revealed ferric ions that stained positively with Prussian blue. In the control group, there is no positive reaction.CONCLUSION:Prussian blue staining in the diagnosis of ocular siderosis has a very significant worth,suspected cases can be definitive diagnosed.

Preparation, optical properties and cell staining of water soluble amine-terminated PAMAM G2.0-Au nanocomposites

The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0（NH2-PAMAM G2.0）-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of Au（Ⅲ） to NH2-PAMAM G2.0 were studied by both UV-visible spectrometry and fluorospectrometry. The NH2-PAMAM G2.0-Au nanocomposites, with a type of structure in which one Au nanoparticle is surrounded by several NH2-PAMAM G2.0 dendrimers, emit strong bluish violet fluorescence, and are uniform, water soluble and biocompatible as well as very stable in frozen conditions. The size of gold nanoparticles in the nanocomposites is about 2.5 nm and decreases with the increase of NH2-PAMAM G2.0 concentration. The NH2-PAMAM G2.0 plays an important role in acting as host or micro-reactor for Au（Ⅲ） before Au（Ⅲ） reduction and acting as dispersant and stabilizer for gold nanoparticles after Au（Ⅲ） reduction. Preliminary experiments of cells staining to human embryonic lung fibroblast cell lines show that the NH2-PAMAM G2.0-Au nanocomposites can be used as optical imaging markers for bioanalyses and medical diagnoses.

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.

Pollen viability of Polygala paniculata L.(Polygalaceae)using different staining methods

Polygala paniculata L.is a medicinal plant that grows in the Brazilian Atlantic coast,known as‘barba-de-São-João’,‘barba-de-bode’,‘vassourinha branca’,and‘mimosa’.In this study,pollen viability was estimated by three different staining methods:2%acetic orcein,2%acetic carmine,and Alexander’s stain.The young inflorescences of twenty accessions were collected and fixed in a solution of ethanol:acetic acid(3:1)for 24 hours,then stored in ethanol 70%under refrigeration.Six slides per plant,two for each stain,were prepared by squashing,and 300 pollen grains per slide were analyzed.Pollen viability was high(>70%)for most accessions of P.paniculata using the Alexander’s stain,which proved the most adequate method to estimate pollen viability.