Changes in activity of layer 2/3 cortical neurons in isoflurane-anesthetized mice: Real-time Ultra-large-Scale High-resolution imaging platform / 中华麻醉学杂志

Objective:To observe and analyze the changes in activity of layer 2/3 cortical neurons in isoflurane-anesthetized mice by Real-time Ultra-large-Scale High-resolution (RUSH) imaging platform.Methods:Clean-grade healthy male Rasgrf2-Cre/Ai148d mice, aged 8-12 weeks, weighing 18-25 g, were studied.The mice recovered ten days after the skull replacement surgery and proceeded to the next experiment.Imaging data of calcium fluorescence signals from layer 2/3 cortical neurons were acquired by RUSH imaging platform after fixing the head of mice.The time of imaging data acquisition in the awake state, during anesthesia with 1.2% isoflurane, and after the end of anesthesia was 100, 600 and 600 s, respectively.Imaging data were analyzed using Image J and MATLAB softwares.Results:The overall trend of activity of layer 2/3 cortical neurons decreased first and then stabilized with the inhalation of 1.2% isoflurane.The cortical neural activity were gradually increased when isoflurane inhalation was stopped.The recovery rate of neural activity was different in different brain regions after isoflurane inhalation was stopped.The recovery of neural activity in the primary motor cortex was delayed obviously.During the maintenance of anesthesia, the activities of most layer 2/3 cortical neurons in the retrosplenial cortex were weakened, however, some of the neurons became more active.Conclusions:The neural activity in the 2/3 layer of cortex in isoflurane anesthetized mice is inconsistent in observation region, brain region and single cell, suggesting that different neural pathways are involved in the process of anesthesia induction and recovery from anesthesia.

Efficacy of modified Parks incision in the surgery for correction of strabismus in children / 中国基层医药

Objective:To investigate the clinical efficacy of modified parks incision in the surgery for correction of strabismus in children.Methods:Sixty children patients with strabismus who received treatment in Shanxi Hospital of Integrated Traditional and Western Medicine, China between January 2015 and January 2020 were included in this study. They were randomly assigned to receive surgery with either a trapezoidal flap incision (control group, n = 30) or modified Parks incision (modified Parks group, n = 30). Time to tear film break-up, amount of tears secreted, and the angles of deviation in strabismus were compared before and after surgery. The efficacy was evaluated by corneal fluorescein staining score. Postoperative complications were compared between the two groups. Results:Before surgery, there were no significant differences in the time to tear film break-up and the amount of tears secreted between the two groups (both P > 0.05). After surgery, the time to tear film break-up and the amount of tears secreted were (9.16 ± 1.74) seconds, (7.51 ± 1.36) mm/5 minutes in the modified Parks group and they were (7.57 ± 1.45) seconds and (6.05 ± 1.14) mm/5 minutes, respectively in the control group. After surgery, the time to tear film break-up and the amount of tears secreted in each group were shortened or decreased compared with before treatment, and these two indices in the modified Parks group were significantly shorter or less than those in the control group ( t = 3.845, 4.506, both P < 0.05). Before surgery, there were no significant differences in corneal fluorescein staining score and the angles of deviation in strabismus between the two groups (both P > 0.05). After surgery, corneal fluorescein staining score and the angles of deviation in strabismus in the modified Parks group were (5.14 ± 1.51) points and (10.68 ± 1.75) PD, respectively and they were (6.25 ± 1.73) points and (15.95 ± 2.14) PD, respectively in the control group. After surgery, corneal fluorescein staining score or the angle of deviation in strabismus was increased or decreased in each group. The corneal fluorescein staining score and the angle of deviation in strabismus in the modified Parks group were significantly lower than those in the control group ( t = 2.648, 10442, both P < 0.05). Total effective rate in the modified Parks group was significantly higher than that in the control group [96.7% (29/30) vs. 80.0% (24/30), χ2= 4.043, P < 0.05]. The incidence of postoperative complications and discomfort rate in the modified Parks group were 0.0% (0/30) and 53.3% (16/30), respectively, which were significantly lower than those in the control group 13.3% (4/30) and 80.0% (24/30), χ2 = 4.286, 4.800, both P < 0.05]. Conclusion:Modified Parks incision for corrective strabismus surgery can better correct the angle of strabismus, protect the stability of tear film function, reduce postoperative complications and decrease postoperative discomfort rate compared with trapezoidal flap incision.

Preliminary clinical application of fluorescence microscopic imaging and computer-aided diagnosis system in the diagnosis of superficial cutaneous fungal infections / 中华皮肤科杂志

Objective@#To evaluate the accuracy of automated fluorescence microscopic imaging and computer-aided diagnosis system (AFMICADS) in the auxiliary diagnosis of superficial cutaneous fungal infections.@*Methods@#Totally, 106 outpatients and inpatients with suspected superficial fungal infections were enrolled from clinical departments of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between July 2018 and September 2018. A total of 126 specimens were collected, including 83 skin scales and 43 nail parings. Each specimen was divided into 3 groups to be examined by conventional fungal microscopy, culture with modified Sabouraud dextrose agar and fluorescence microscopy (artificial fluorescence microscopy and AFMICADS-based fluorescence microscopy) respectively. A positive result was defined as that conventional fungal microscopy and/or fungal culture was positive. Consistency rate, sensitivity and specificity of the 3 microscopic methods were calculated. Statistical analysis was carried out with SPSS 10.0 software by using McNemar test and Kappa test for analyzing difference in the positive rate, as well as consistency, between the 3 microscopic methods and the positive standard, and by using efficiency test for comparing the consistency rate among the 3 microscopic methods.@*Results@#Of 126 specimens, 124 (98.4%) were positive for artificial fluorescence microscopy, and 123 (97.6%) for AFMICADS-based fluorescence microscopy. Both positive rates of the above 2 microscopic methods were significantly higher than the positive rate of the positive standard (77.8%, both P < 0.001) . The sensitivity, specificity and consistency rate of AFMICADS-based fluorescence microscopy were 100%, 10.7% and 80.2% respectively, and those of artificial fluorescence microscopy were 100%, 7.1% and 79.4% respectively. Additionally, no significant difference in the consistency was observed between the AFMICADS-based and artificial fluorescence microscopy (P > 0.05) .@*Conclusion@#The accuracy of AFMICADS-based fluorescence microscopy in the diagnosis of superficial cutaneous fungal infections is similar to that of artificial fluorescence microscopy.

Preliminary clinical application of fluorescence microscopic imaging and computer-aided diagnosis system in the diagnosis of superficial cutaneous fungal infections / 中华皮肤科杂志

Objective To evaluate the accuracy of automated fluorescence microscopic imaging and computer-aided diagnosis system (AFMICADS) in the auxiliary diagnosis of superficial cutaneous fungal infections.Methods Totally,106 outpatients and inpatients with suspected superficial fungal infections were enrolled from clinical departments of Union Hospital,Tongji Medical College,Huazhong University of Science and Technology between July 2018 and September 2018.A total of 126 specimens were collected,including 83 skin scales and 43 nail parings.Each specimen was divided into 3 groups to be examined by conventional fungal microscopy,culture with modified Sabouraud dextrose agar and fluorescence microscopy (artificial fluorescence microscopy and AFMICADS-based fluorescence microscopy) respectively.A positive result was defined as that conventional fungal microscopy and/or fungal culture was positive.Consistency rate,sensitivity and specificity of the 3 microscopic methods were calculated.Statistical analysis was carried out with SPSS 10.0 software by using McNemar test and Kappa test for analyzing difference in the positive rate,as well as consistency,between the 3 microscopic methods and the positive standard,and by using efficiency test for comparing the consistency rate among the 3 microscopic methods.Results Of 126 specimens,124 (98.4％) were positive for artificial fluorescence microscopy,and 123 (97.6％) for AFMICADS-based fluorescence microscopy.Both positive rates of the above 2 microscopic methods were significantly higher than the positive rate of the positive standard (77.8％,both P ＜ 0.001).The sensitivity,specificity and consistency rate of AFMICADS-based fluorescence microscopy were 100％,10.7％ and 80.2％ respectively,and those of artificial fluorescence microscopy were 100％,7.1％ and 79.4％ respectively.Additionally,no significant difference in the consistency was observed between the AFMICADS-based and artificial fluorescence microscopy (P ＞0.05).Conclusion The accuracy of AFMICADS-based fluorescence microscopy in the diagnosis of superficial cutaneous fungal infections is similar to that of artificial fluorescence microscopy.

Biomateriais com aplicabilidade na ortopedia: avanços e desafios na área da infectologia / Biomaterials with applicability in orthopedics: advances and challenges in the infectology area

O controle na formação do biofilme em implantes e próteses ortopédicas continua sendo um dos grandes desafios acerca da infeção relacionada aos dispositivos na área da saúde. O objetivo desta pesquisa foi investigar biomateriais com aplicabilidade na ortopedia, visando os avanços e enfrentamentos dos desafios na área da infectologia. Uma revisão integrativa foi realizada a respeito da formação de biofilme em biomateriais de próteses de quadril com a finalidade de contribuir com as medidas de prevenção e controle aos agravos infecciosos. Além disso, a formação in vitro do biofilme em função dos biomateriais (titânio e titânio revestido com biovidro F18), microrganismos (Staphylococcus epidermidis e Candida albicans) e tempos de incubação (2, 4 e 8 horas) foi avaliada por microscopia de fluorescência. A revisão integrativa foi realizada no portal PubMed da National Library of Medicine, bem como nas bases Cochrane, Embase, Web of Science, CINAHL e LILACS com a inclusão de estudos primários sobre a temática, publicados online até novembro de 2017, em português, inglês e espanhol. Na fase experimental / laboratorial, biofilmes de S. epidermidis (ATCC 12228) e C. albicans (ATCC 90028) foram formados em corpos de prova de titânio e titânio revestido com biovidro F18 após 2, 4 e 8 horas de incubação a 37?C sob agitação orbital. As áreas das imagens dos corpos de prova, em porcentagem, recobertas com biofilme (células vivas) foram avaliadas por microscopia de fluorescência. Os dados coletados foram submetidos à análise estatística empregando-se os testes de normalidade Shapiro Wilk, U de Mann-Whitney e t de Student por meio do software IBM SPSS Statistics (versão 25) e nível de significância ?=5%. Na revisão integrativa, os resultados demonstraram que dos 16 estudos primários, 81,25% eram pesquisas experimentais in vitro e que novos biomateriais foram desenvolvidos para prevenir a formação de biofilme. Com relação à fase experimental / laboratorial, houve menor formação de biofilme por S. epidermidis e C. albicans (p<0,001) no titânio revestido com biovidro F18 do que no titânio, após 8 horas de incubação. Entretanto, houve maior formação de biofilme por S. epidermidis e C. albicans após 8 horas do que em 2 horas de incubação, tanto no titânio quanto no titânio revestido com biovidro F18 (p<0,05). Em suma, a revista da literatura mencionou o desenvolvimento de biomateriais novos para prevenir a formação de biofilme. Na fase laboratorial / experimental, o titânio revestido com biovidro F18 apresentou atividade antibiofilme em comparação com o titânio, e os tempos de incubação de 2 para 8 horas aumentaram a formação de biofilme em ambos os biomateriais. Ainda, pesquisas futuras acerca do biovidro F18 fundamentadas nos aspectos físicoquímicos, bioquímicos e microbiológicos são importantes para a elucidação dos mecanismos de ação relacionados ao controle dos biofilmes

The control of biofilm formation on implants and orthopedic prostheses still is one of the major challenges concerning infection related to devices in the health field. The objective of this research was to investigate biomaterials with applicability in orthopedics, aiming for advances and facing challenges in the infectology area. An integrative review was performed regarding biofilm formation on hip prosthesis biomaterials in order to contribute to the preventive and infection control measures. Moreover, the in vitro biofilm formation according to biomaterials (titanium and titanium coated with F18 bioglass), microorganisms (Staphylococcus epidermidis and Candida albicans) and incubation times (2, 4 and 8 hours) was evaluated by fluorescence microscopy. The integrative review was performed on PubMed portal from National Library of Medicine as well as on Cochrane, Embase, Web of Science, CINAHL and LILACS databases with the inclusion of primary studies about the topic, published online up until November 2017, in Portuguese, English and Spanish. In the experimental / laboratory step, S. epidermidis (ATCC 12228) and C. albicans (ATCC 90028) biofilms were formed on proof bodies of titanium and titanium coated with F18 bioglass after 2, 4 and 8 hours of incubation at 37?C under orbital shaking. The image areas of proof bodies, in percentage, coated with biofilm (living cells) were evaluated by fluorescence microscopy. The data collected were submitted to statistical analysis using normality tests Shapiro Wilk, U from Mann-Whitney and t from Student through IBM SPSS Statistics (version 25) software and significance level ?=5%. In the integrative review, the results showed that among 16 primary studies, 81.25% were in vitro experimental studies and that new biomaterials were developed to prevent biofilm formation. Regarding experimental / laboratory step, there was less biofilm formation by S. epidermidis and C. albicans (p<0.001) on titanium coated with F18 bioglass than on titanium, after 8 hours of incubation. However, there was more biofilm formation by S. epidermidis and C. albicans after 8 hours than in 2 hours of incubation, both on titanium and on titanium coated with F18 bioglass (p<0.05). In sum, the literature review mentioned the development of new biomaterials to prevent biofilm formation. In laboratory / experimental step, titanium coated with F18 bioglass presented antibiofilm activity in comparison with titanium, and the incubation times of 2 to 8 hours increased biofilm formation on both materials. Besides, future studies about F18 bioglass based on physicochemical, biochemical and microbiological aspects are important for the elucidation of action mechanisms related to biofilms control

Clinical and histological features of patients with membranoproliferative glomerulonephritis classified by immunofluorescence findings / Características clínicas e histológicas de pacientes com glomerulonefrite membranoproliferativa classificados por achados de imunofluorescência

Abstract Background: New classification for membranoproliferative glomerulonephritis has been proposed in the literature. The aim of this study was to compare the clinical, biochemical, etiology and renal biopsy findings of these patients grouped by immunofluorescence as proposed by the new classification. Methods: Patients with renal biopsy-proven membranoproliferative glomerulonephritis unrelated to systemic lupus erythematosus, diagnosed between 1999 and 2014. The patients were divided according to immunofluorescence: Immunoglobulin positive group, C3 positive only and negative immunofluorescence group. Results: We evaluated 92 patients, the majority of which were in the immunoglobulin positive group. Infectious diseases, hepatitis C virus and schistosomiasis, were the most frequent etiology. A negative immunofluorescence group had more vascular involvement in renal biopsy compare with others groups. Conclusions: The only difference between the groups was higher vascular involvement in renal biopsy in negative immunofluorescence group. These new classification was satisfactory for the finding of etiology in one part of the cases.

Resumo Introdução: Uma nova classificação para glomerulonefrite membranoproliferativa foi proposta na literatura. O objetivo deste estudo foi comparar os achados clínicos, bioquímicos, etiológicos e da biópsia renal desses pacientes agrupados por imunofluorescência, conforme proposto pela nova classificação. Métodos: Pacientes com glomerulonefrite membranoproliferativa comprovada por biópsia renal, não relacionada ao lúpus eritematoso sistêmico, diagnosticados entre 1999 e 2014. Os pacientes foram divididos de acordo com a imunofluorescência: grupo positivo por imunoglobulina, grupo positivo por C3 apenas e grupo com imunofluorescência negativa. Resultados: avaliamos 92 pacientes, a maioria dos quais estava no grupo de imunoglobulina positiva. Doenças infecciosas, o vírus da hepatite C e a esquistossomose, foram as etiologias mais frequentes. Um grupo com imunofluorescência negativa apresentou maior comprometimento vascular na biópsia renal quando comparado com os outros grupos. Conclusões: a única diferença entre os grupos foi o maior envolvimento vascular na biópsia renal no grupo de imunofluorescência negativa. Esta nova classificação foi satisfatória para a descoberta de etiologia em uma parte dos casos.

The application of acridine orange fluorescent staining method in circulating tumor cell / 天津医药

Objective To establish the screening platform of circulating tumor cells (CTCs) using acridine orange fluo?rescent (AO-F) dyeing method, and to apply it in the screening of peripheral blood CTCs in patients with kidney cancer. Methods Twenty-seven patients with metastatic renal cell carcinoma was included in this study. Primitive tumor cells and kidney cancer cell line 769-P were cultured with different concentrations of fetal bovine serum. Smears were prepared and observed under fluorescence microscopy. The percentage of AO-F positive staining of 769-P cells under 5 random sights was calculated. The sensitivity of AO-F staining to cells was evaluated. The 5 mL morning fasting venous blood was obtained from 10 subjects with healthy check-up. The 1×106 cell suspension was prepared. The logarithmic phase of renal tumor cells was used to prepare tube containing 500, 200, 100, 50 and 10 tumor cell suspension, which were mixed with 1×106 nucleated cells to establish CTCs model of renal cancer. AO-F staining method was used to detect the expression of AO-F positive cells. The correlation between expression of AO-F positive cells and clinical parameters was analyzed. Results The prima?ry cells and cell line 769-P showed similar bright color and morphological characteristics. The percentage of AO-F positive staining in 769-P cells was 93%±3%under 5 random sights. The recovery rates (%) of four groups (500, 200, 100 and 50 tu?mor cell suspension) were 10.2±3.8, 9.2±2.3, 10.8±2.6 and 10.5±1.9, respectively. There were no significant differences in recovery rates between four groups (P>0.05). The group of 10 tumor cell suspension could find AO-F positive staining cells occasionally. Zero case was positive in controls. Nine of 27 patients were positive and the rate was 33.33%. There were no significant statistical differences in AO-F positive rates between gender, age, tumor size, pathological pattern, Furhman stage, metastasis of lung and presence of tumor (P>0.05). Conclusion It is confirmed that the method of CTCs staining with AO-F, which has high specificity and reproducibility, is feasible to detect CTCs and worthy of being studied. There is a certain reference value to predict tumor recurrence and metastasis.

Osteogenetic ability of mouse spermatogonial stem cells cultured in vitro / 中国组织工程研究

BACKGROUND:Spermatogonial stem cells are a kind of adult stem cells, which have self-renewal and differentiation potential, and can be differentiated into specific cells in vitro, suggesting that the spermatogonial stem cells may be possibly differentiated into osteoblasts. But the related research has not been reported. OBJECTIVE:To observe the biological characterization and osteogenic process of mouse spermatogonial stem cells cultured in vitro. METHODS:Spermatogonial stem cells were obtained from the testicle of mice aged 15-20 days, and were cultured on the feeder layer from bone marrow stroma cells in vitro. When cultured for 3 days, the cells were cultured in the conditioned medium (experimental group) and basic medium (control group). The cells proliferation capability and osteogenic property were examined by phase-contrast microscope, alkaline phosphatase activity and type I col agen immunofluorescence staining. RESULTS AND CONCLUSION:Spermatogonial stem cells proliferated faster in the experiment group than in the control group. cells grew rapidly in colony-like shape in the conditioned medium at 3-6 days, the three-dimensional feeling enhanced, cellmass and clusters continued to increase in size, the extracellular matrix was increased in number and the cytoplasmic bridge was not obvious. After culture for 15 days, cells in the two groups were positive for alkaline phosphatase staining that the cytoplasmic membrane was dyed black. Under the fluorescent microscope, green fluorescence was visible in the experimental group, suggesting the cells in the experimental group was positive for type I col agen, but negative in the control group, which is similar with the biological characteristics of osteoblasts. These findings indicate that spermatogonial stem cells possess the osteogenic capability under induction conditions, which are expected to provide seed cells for bone tissue engineering.

Expression and distribution of aquaporin 3 during early embryonic development of mouse / 中华围产医学杂志

Objective To investigate the expression and distribution of aquaporin 3 (AQP3) in mouse early embryos at different stages.Methods Controlled ovarian hyperstimulation model of Kunming mouse was used to collect four-cell embryos,eight-cell embryos,morula stage,and early blastocysts.Immunofluorescence microscopy and laser confocal microscopy were used to detect expression and distribution of AQP3 channels in these stages.Results Fluorescence signal of AQP3 was found in four embryonic stages of mice.Distribution within embryo was different at different embryonic stages.AQP3 was mainly expressed on the karyotheca of blastomeres at four-cell and eight-cell stage.In morula stage,AQP3 was mainly expressed on cell membrane of each blastomere.In early blastocysts,AQP3 was predominantly expressed on the cell membrane and cytoplasm of trophoblastic cell.Conclusions AQP3 trans-membrane channel might have potential regulation function on mouse embryonic development.

Coma blisters after poisoning caused by central nervous system depressants: case report including histopathological findings / Bolhas do coma pós-intoxicação por depressores do sistema nervoso central: relato de caso incluindo achados histopatológicos

Blister formation and eccrine sweat gland necrosis is a cutaneous manifestation associated with states of impaired consciousness, most frequently reported after overdoses of central nervous system depressants, particularly phenobarbital. The case of a 45-year-old woman who developed "coma blisters" at six distinct anatomic sites after confirmed (laboratory) phenobarbital poisoning, associated with other central nervous system depressants (clonazepam, promethazine, oxcarbazepine and quetiapine), is presented. A biopsy from the left thumb blister taken on day 4 revealed focal necrosis of the epidermis and necrosis of sweat gland epithelial cells; direct immunofluorescence was strongly positive for IgG in superficial blood vessel walls but negative for IgM, IgA, C3 and C1q. The patient was discharged on day 21 with no sequelae.

Formação de bolhas e necrose de glândula sudoríparas écrinas é uma manifestação cutânea associada com estados de diminuição da consciência, mais frequentemente relatada após superdosagens de depressores do sistema nervoso central, particularmente fenobabital. Relatamos o caso de uma paciente de 45 anos que desenvolveu "bolhas do coma" após tentativa de suicídio por fenobarbital (confirmada laboratorialmente), associada a outros depressores do sistema nervoso central (clonazepam, prometazina, oxcarbazepina e quetiapina). Biópsia da bolha do 1o quirodáctilo esquerdo no 4o dia de internação revelou necrose focal da epiderme e necrose de células epiteliais de glândula sudorípara; a imunofluorescência direta foi fortemente positiva para IgG na parede superficial dos vasos sanguíneos, mas negativa para IgM, IgA, C3 e C1q. A paciente teve alta no 21o dia, sem seqüelas.

Comparação entre três métodos de coloração a frio no diagnóstico primário de tuberculose: um estudo piloto / Comparison among three cold staining methods in the primary diagnosis of tuberculosis: a pilot study

OBJETIVO: Em países em desenvolvimento, a baciloscopia é a principal ferramenta para a identificação de casos de tuberculose pulmonar. O objetivo do presente estudo foi avaliar a eficácia diagnóstica do método de coloração de Gabbett (MCG) e de um método modificado de coloração a frio (MMC), ambos em duas etapas, em comparação com a do método de coloração fluorescente (MCF), em três etapas, para a detecção de BAAR em esfregaços de escarro. MÉTODOS: Nossa amostra consistiu de 260 amostras de escarro coletadas de casos suspeitos de tuberculose pulmonar no Kasturba Hospital, em Manipal, Índia. Os esfregaços foram preparados em triplicata, para cada um dos métodos: MCF, MMC e MCG. As lâminas foram numeradas aleatoriamente a fim de que o examinador fosse cegado quanto à identidade das amostras. RESULTADOS: Das 260 amostras, 16 (6,15 por cento), 15 (5,77 por cento) e 13 (5,00 por cento) foram positivas para BAAR com MCF, MMC e MCG, respectivamente. A sensibilidade de MCG e MMC em relação à de MCF foi de 81,25 por cento e 93,75 por cento, respectivamente. Houve boa concordância de MCG e MMC com MCF (0,988 e 0,996, respectivamente), e não houve diferenças estatísticas significativas. CONCLUSÕES: Embora MCG e MMC apresentaram menor sensibilidade que MCF, que é avaliado por microscopia de fluorescência, consideramos que os dois primeiros métodos sejam promissores no diagnóstico de tuberculose.

OBJECTIVE: In developing countries, sputum smear microscopy is the main tool for pulmonary tuberculosis case finding. The objective of the present study was to evaluate the diagnostic efficacy of Gabbett's staining (GS) and modified cold staining (MCS), both of which are two-step methods, in comparison with that of fluorescent staining (FS), which is a three-step method, for the detection of AFB in sputum smears. METHODS: Our sample comprised 260 sputum samples collected from individuals suspected of having pulmonary tuberculosis at Kasturba Hospital, in Manipal, India. Smears were prepared in triplicate: one each for FS, MCS, and GS. The smears were randomly numbered so that the examiner was blinded to the sample identities. RESULTS: Of the 260 samples, 16 (6.15 percent), 15 (5.77 percent), and 13 (5.00 percent) showed positive AFB results with FS, MCS, and GS, respectively. The sensitivity of GS and MCS, in comparison with that of FS, was 81.25 percent and 93.75 percent, respectively. The concordance of GS and MCS with FS was good (0.988 and 0.996, respectively), and no statistically significant differences were found. CONCLUSIONS: Although MCS and GS were found to be less sensitive than was FS, which is evaluated under fluorescence microscopy, the first two are promising methods for the diagnosis of tuberculosis.

Porfiria cutânea tardia: relato de dois casos / Porphyria cutanea tarda: case two reports

JUSTIFICATIVA E OBJETIVOS: A infrequência da porfiria cutânea tardia (PCT) e as suas várias formas de apresentação clínica justificam tal exposição. O objetivo deste estudo foi relatar dois casos de PCT: uma secundária a fármacos e outra de origem familiar. RELATO DO CASO: Caso 1: Paciente do sexo feminino, 70 anos com lesões bolhosas e vesiculosas no dorso das mãos, face e couro cabeludo havia dois anos, além de cicatrizes com milium. Fazia uso de fenitoína para epilepsia, sendo a PCT desencadeada por este fármaco. Caso 2: Trata-se de uma paciente de 35 anos com lesões vesiculosas e bolhosas no dorso das mãos, úlceras de difícil cicatrização, hipercromia e hipertricose facial. História familiar positiva de um irmão com hemocromatose em tratamento.CONCLUSÃO: A PCT resulta de um defeito enzimático caracterizado por diminuição da atividade da uroporfirinogênio-decarboxilase. Hemocromatose hereditária, hepatite C, estrógenos e história familiar constituem os principais fatores de risco para porfiria cutânea tardia. Clinicamente, observam-se vesículas, bolhas e ulcerações em áreas fotoexpostas, cicatrizes, pequenos cistos (milium), hipertricose e hiperpigmentação cutânea. O diagnóstico é clínico associado à dosagem de porfirinas na urina, sangue e fezes, podendo ser complementado com o exame histopatológico. A terapêutica inclui a eliminação do fator causal, flebotomias repetidas e/ou administração de cloroquina ou cimetidina por via oral.

BACKGROUND AND OBJECTIVES: The porphyria cutanea tarda (PCT) is an infrequent dermatosis and has several presentations that justify this paper. This objective of study was relating two PCT cases: one of them is a secondary drugs form and the other is a familiar one. CASE REPORT: Case 1: A woman patient, 70 old year with bullous and vesicles on the hand, face and scalp along two years beside milium scars. This patient was being treated with phenytoin for brain?s cysticercosis and this drug probably desencaded the PCT. The urinary?s porphyrinas dosage elevated associated with histopalogic confirmed the diagnostic. Case 2: Woman patient, 35 old year with bullous and vesicles on the hand, ulcers that not scars. There was facial hypercromia and hypertrichosis. Hers brothers has hemocromatosis. CONCLUSION: The PCT is an enzymatic defect characterized for reduced uroporphyrinogen decarboxilasi?s activity. Hereditary hemocromatosis, C hepatitis, estrogens and family history are the main risk factors for porphyria cutanea tarda. Clinically, we can observe vesicles, bullae and ulcers in areas exposed to light, atrophic scars, small cysts (milia), hypertrichosis and cutaneous hyper pigmentation. Clinical aspects associated to porphyrina's detection urinary, blood and stools makes the diagnosis. It can be complemented by skin's histopathology. The treatment can be done by eliminating the risk factors, repeated phlebotomies and/or introducing oral chloroquine or cimetidine.

Venous drainage in the lesser saphenous sural neurovenofasciocutaneous distally based flap:an experimental study in rabbit model / 中华创伤杂志

Objective To investigate the venous drainage of lesser saphenous aural neurov enofasciocutaneous distally based flap through fluorescence tracing technique and discuss the pattern of venous drainage.Methods Venous blood for 0.1 ml was collected from every rabbit ear vein of 20 rabbits respectively for separation of the erythrocytes and labeling with FITC.The lesser saphenous sural neurovenofasciocutaneous distally based flaps were successfully established in hind limbs of 20 rabbits that were then allocated into four groups according to different inspection time points at 30 minutes ( Croup A) ,24 hours (Group B) ,72 hours (Group C) and 7 days (Group D) after operation.The labeled erythrocytes (5 μl) were injected into the flaps via lesser saphenous vein (in Groups A and B)or hypoderma (in Groups C and D).Then,the flaps were removed five seconds (in Groups A and B) or 10 seconds (in Groups C and D) after injection,immediately frozen and sectioned (5-7 μm in thickness) for microscopic analysis of fluorescent distribution in the pedicle.Results FITC-labeled red blood cells showed steady green fluorescence under inversion fluorescence microscope.Fluorescence was mainly distributed in the wall of lesser saphenous vein and peripheral vessels,as well as inner and outer membrane of perforator artery.There was only faint fluorescence around sural nerve in Groups B,C and D.HE staining showed that the lumina of lesser saphenous vein in Groups C and D were fully filled with thrombosis.Conclusions Vein of the lesser saphenous sural neurovenofasciocutaneous distally based flaps is refluxed mainly through wall of lesser saphenous vein and peripheral vessels as well as through inner and outer membrane of perforator artery in the pedicle.Thrombosis occurs in the lumina of lesser saphenous but there is no venous blood reflux through the valve of lesser saphenous vein.

Detection of homodimer formation of CD99 through extracelluar domain using bimolecular fluorescence complementation analysis

Although various functions of CD99 have been reported, such as apoptosis and homotypic aggregation of thymocyte and transendothelial migration of immune cells, biochemical/molecular natures of CD99 are still elusive. Using mouse CD99 gene, we show that CD99 forms homodimer through its extracellular domain. Expression of mouse CD99 is up-regulated on T cells after CD3-mediated activation, like the case for human CD99. The potential of CD99 to form homodimer was tested with a recently developed bimoleular fluorescence complementation analysis (BiFC). In BiFC analysis, the dimerization-induced fluorescence was strong near the perinuclear region and was faded at the cell membrane. However, surface expression of CD99 was still detected by flow cytometry, suggesting that CD99 either in monomer form or in association with other molecules exists on the cell surface. In BiFC analysis using CD99 mutants with its extracellular, transmembrane, or cytosolic domains changed to corresponding human CD4 domains, the mutant replaced with human CD4-extracellular domain did not produce fluorescence. Purified soluble CD99-Fc fusion proteins bound to CD99-Fc immobilized onto the gold sensor chip in surface plasmon resonance analysis, confirming that the extracellular domain was responsible for dimer formation. Intracytoplasmic staining for CD99 expression in the thymocytes and mature T cells showed that most of the cells, even the cells with low surface level of CD99, contained the molecule inside the cell. Our results suggest that majority of CD99 homodimers may exit in the cell and be exported to the cell surface, dissociating from each other, after a certain regulatory signal is delivered.