Myoid differentiation in dermatofibrosarcoma protuberans and its fibrosarcomatous variant: 10 years' experience in a tertiary hospital.

Dermatofibrosarcoma protuberans (DFSP) is a relatively rare, locally aggressive, and dermal-based fibroblastic tumor. There are several histological variants, in which the usual emphasis is on fibrosarcomatous DFSP, as it acquires metastatic potential. Myoid differentiation in DFSP is rare, and more often found in fibrosarcomatous DFSP. Myoid differentiation is defined as tumor cells with brightly eosinophilic cytoplasm, well-defined cytoplasmic margins, and vesicular nuclei. In this study, we aim at characterizing the immunostaining pattern regarding myoid differentiation in DFSP, and discuss the potential pitfall in making the diagnosis. A total of ten cases of DFSP were found in the past ten years in our hospital. Two of them show focal myoid differentiation, including the only case of fibrosarcomatous DFSP. Around 5% of the tumor area in the traditional DFSP case shows myoid differentiation, while around 10% of the tumor area in fibrosarcomatous DFSP shows myoid differentiation. The myoid areas show positive staining, albeit patchy to focal, for smooth muscle markers, including smooth muscle actin, muscle-specific actin, caldesmon, and calponin. Staining for CD34, in those areas, is weak or negative. This may create diagnostic difficulty with smooth muscle tumors or myofibroblastic lesions, especially in a small biopsy sample. In difficult cases, the detection of COL1A1-PDGFB fusion by fluorescence in situ hybridization is helpful, as this is a characteristic chromosomal translocation found in the large majority of DFSP.

Protective effects of acupuncture and LGNHFD on expressions of vascular endothelial growth factor, basic fibroblast growth factor, and cluster of differentiation 34 in rats with cerebral ischemia-reperfusion injury.

OBJECTIVE: To investigate the protective effects of electroacupuncture and LGNHFD on cerebral ischemia-reperfusion injury, and their effects on the expression of cluster of differentiation 34 (CD 34), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) in the peripheral tissue of the cerebral ischemic semi-dark zone in rats. The influence and mechanism of expression. METHODS: Healthy male Sprague-Dawley rats were randomly divided into the sham operation group, model group, Traditional Chinese Medicine (TCM) group, electroacupuncture group, and acupuncture group. The cerebral ischemia model was prepared by middle artery occlusion. The electroacupuncture and TCM groups received electroacupuncture at the ""Neiguan (PC6)"", ""Baihui (GV20)"", and ""Renzhong (GV26)"" acupoints from 4 h after modeling, once daily for 14 d. After modeling, the acupuncture and TCM groups were administered LGNHFD once daily for 14 d. After treatment, the infarct volume and regional cerebral blood flow (rCBF) were measured in each group, the expression of CD34 in the surrounding ischemic penumbra was determined using immunohistochemistry, and the expression of angiogenesis-related factors in the ischemic hemisphere was detected by western blot. RESULTS: Compared with the sham operation group, rCBF was significantly decreased (P < 0.01) and cerebral infarction volume was significantly increased (P < 0.01) in the model group at four time points (5 min and 3, 7, and 14 d after modeling). At three postoperative time points (3, 7, and 14 d), the number of CD34+ cells in the model group were significantly increased (P < 0.01), and the relative expression levels of VEGF, bFGF, and CD34+ in the ischemic hemisphere were significantly increased (P < 0.01, P < 0.05). Compared with the model group, the rCBF of the acupuncture group was significantly increased at 3, 7, and 14 d after intervention (P < 0.01), while the rCBF of the electroacupuncture and TCM groups was significantly increased at 7 and 14 d after intervention (P < 0.01). After 3, 7, and 14 d of intervention, infarct volume was significantly reduced (P < 0.01), the number of CD34+ cells was significantly increased (P < 0.01, P < 0.05), and the relative expression levels of VEGF, bFGF, and CD34 in the ischemic hemisphere were significantly increased (P < 0.01, P < 0.05). Compared with the electroacupuncture and TCM groups, the rCBF of the acupuncture group was significantly increased (P < 0.01), the infarct volume was significantly reduced (P < 0.01), and the relative expression levels of VEGF, bFGF, and CD34 were significantly increased (P < 0.05, P < 0.01) after 3, 7, and 14 d of intervention. CONCLUSION: Acupuncture therapy is superior to simple drug and electroacupuncture therapy for reducing infarct volume, increasing rCBF and CD34 expression in cells, and promoting capillary regeneration in rats with cerebral ischemia. This may be related to the increased expression of VEGF, bFGF, and CD34 - which are related to cerebrovascular neovascularization - in the ischemic hemispheres of such rats.

Involvement of tryptase-positive mast cells and angiogenesis in the growth of inflammatory odontogenic cysts

Abstract: Inflammatory periapical lesions are characterized by infiltration of different immune cell types, the functions of which depend on an effective vascular network. This study aimed to evaluate the mast cells density (MCD) in inflamatory odontogenic cysts capsules concerning microvascular density (MVD), microvascular area (MVA), and microvascular perimeter (MVP), and correlate such findings with the type of lesion, intensity of the inflammatory infiltrate, and thickness of the epithelial lining. Twenty inflamatory dentigerous cysts (IDCs), twenty radicular cysts (RCs), and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using anti-tryptase and anti-CD34 antibodies. RCs exhibited the highest MCD, MVD, MVA, and MVP indexes (p = < 0.001, p = 0.008, p = 0.003 and p = < 0.001, respectively), and lesions with inflammatory infiltrate grade III showed the highest MVD (p = 0.044). Considering epithelial thickness, a higher MVP index was identified in lesions with hyperplastic epithelium (p = 0.018). In IDCs, RCs, and RRCs, a strong positive correlation was observed between MVA and MVP (r = 0.950 and p = < 0.001; r = 0.914 and p = < 0.001; r = 0.713 and p = < 0.001, respectively). In IDCs, a moderate correlation was observed between MCD and both MVA and MVP (r = 0.660 and p = 0.002; r = 0.634 and p = 0.003, respectively). These results suggest that tryptase-positive mast cells might play an important role in the angiogenic activity of IDCs, while RCs had the highest indexes. Our findings also confirmed that the intensity of the inflammatory infiltrate and epithelial thickness influence angiogenesis.

Fibromixoma acral superficial envolvendo quirodáctilo: um relato de caso / Superficial acral fibromyxoma involving the fingers: a case report

O fibromixoma acral superficial é um tumor mesenquimal raro e benigno. Acomete principalmente homens de meia-idade; entretanto, pode ocorrer em qualquer sexo e faixa etária. Apresenta crescimento lento, com predileção por áreas ungueais e periungueais.

Superficial acral fibromyxoma is a rare and benign mesenchymal tumor. It mainly affects middle-aged men; however, it can occur in any gender and age group. It has a slow growth, with a preference for nail and periungual areas

[Effects of different time of cryoablation on the proliferation of airway traumatic granuloma and its mechanism in rabbits].

Objective: To investigate the effect of different cryoablation time on tracheal traumatic granulation formation and its mechanism. Methods: A total of 32 rabbits were randomly assigned into four groups (A-D). Group A underwent tracheotomy alone. Group B, C and D received intra-tracheal brush rubbing to establish airway granulation model. Group C and D underwent 30 s and 2-minute cryoablation respectively. Tracheal specimens of all groups were collected to examine pathological changes using HE staining. Levels of transforming growth factor beta 1 (TGF-ß(1)) and CD34 in tracheal granulation were evaluated using immunohistochemistry (IHC) and real-time quantitative reverse transcription polymerase chain reaction (RT-qCR). Results: Tracheal lumens of group A were smooth without granulation. While the growth of granulation and luminal stenosis were most severe in Group B, followed by Group D and C. Submucosa thickness of Group B was largest as compared with other groups (0.20±0.07, 0.77±0.28, 0.44±0.13 and 0.55±0.18 mm for Group A to D, respectively. P<0.05). And the submucosa layer of Group C was thinner than Group D (P<0.05). The expression and transcription levels of TGF-ß(1) of trachea were highest in Group B as detected by IHC and RT-qPCR (P<0.05), followed by Group D and C (IHC: 0.48±0.01 vs 0.43±0.01, P<0.05; RT-qPCR: 12.61±2.14 vs 2.38±0.10, P<0.05). Both protein and mRNA levels of CD34 were highest in Group B as detected by IHC and RT-qPCR (P<0.05). Tracheal mRNA levels of CD34 were more abundant in Group D than Group C (4.92±0.90 vs 2.09±0.10, P<0.05), while no significant difference was found between groups regarding protein levels measured by IHC. Conclusions: Cryoablation could alleviate the hyperplasia of tracheal traumatic granulation, possibly due to the inhibition of TGF-ß(1) and CD34 expression. The effect of 30 s cryoablation for tracheal traumatic granulation is better.

[Clinicopathologic features of atypical spindle cell lipomatous tumor].

Objective: To investigate the clinicopathologic characteristics, immunophenotype, differential and diagnostic features of atypical spindle cell lipomatous tumor (ASLT). Methods: Three cases of ASLT were collected from January 2010 to March 2017 at Zhejiang Provincial People's Hospital. The clinical and imaging features, histomorphology, immunophenotype and prognosis were analyzed. Fluorescence in situ hybridization (FISH) was used to detect MDM2 gene amplification, and relevant literature was reviewed. Results: All three patients were adult males, aged 38, 43 and 54 years, respectively. One tumor originated in the subcutaneous soft tissue in the head and neck, one was located in the left primary bronchus and one in the latissimus dorsi muscle. Grossly, all three tumors were circumscribed and ranged from 4.0 to 5.8 cm in size. Microscopically, all showed a focally infiltrative front. These tumors were composed of variable proportions of spindle-shaped and adipocytic cells in a background of variable fibrous and edematous matrix. Scattered lipoblasts were easily seen. One tumor was composed predominately of spindle tumor cells, one of adipocytic cells, and one of equally mixed cell populations. The spindle tumor cells were generally bland-appearing with focal nuclear enlargement and hyperchromasia noted in one case. Mitosis was not seen in neither the spindle cells nor the adipocytic cells. By immunohistochemistry, diffuse and strong reactivity to CD34 of the spindle cells was noted in all cases, definite loss of Rb expression was noted in one of three cases, and S-100 protein was expressed only in the adipocytic cells. INI-1 was intact and Ki-67 index was 1% to 3%. All other markers including CDK4, MDM2, STAT6, SOX10, CD99, bcl-2, ß-catenin, CD117, GFAP, CK, EMA, SMA and desmin were negative. FISH of MDM2 was done in two cases, and both showed no amplification. The ASLT in the head and neck had two recurrences during 17 months of follow-up, whereas the tumor in the latissimus dorsi was free of disease during 33 months of follow-up. Conclusions: ASLT is a rare subtype of low-grade adipocytic neoplasm and is distinctive from atypical lipomatous tumor/well-differentiated liposarcoma. The histomorpholgy of ASLT has significant heterogeneity and forms a continuous spectrum. ASLT needs to be distinguished from a series of benign and malignant soft tissue tumors.

Effect of plasma exchange on bioactivity of peripheral blood CD34+ cells in patients with acute-on-chronic (subacute) liver failure / 临床肝胆病杂志

ObjectiveTo investigate the effect of improvement in internal environment after plasma exchange on the bioactivity of peripheral blood CD34+ cells in patients with acute-on-chronic (subacute) liver failure. MethodsThe peripheral blood CD34+ cells from patients with acute-on-chronic (subacute) liver failure were cultured in vitro using the medium containing with the serum collected before or after plasma exchange. The growth curves of the two groups of cells were recorded and compared. RT-PCR was used to detect the expression of the cell surface markers PK-M2, Integrin-β1, and L-PK, and immunofluorescence was used to detect the expression of Integrin-β1. A repeated measures analysis of variance was used for comparison of continuous data between groups. ResultsThe CD34+ cells in the medium containing the serum collected after plasma exchange grew better than those in the medium containing the serum collected before plasma exchange (P＜0.05). PK-M2 and Integrin-β1 were detected in the CD34+ cells of both groups, but L-PK was not detected in either group. ConclusionFor patients with liver failure, the improvement in internal environment after plasma exchange helps to maintain the bioactivity of peripheral blood CD34+ cells, thus improving the efficacy of stem cell transplantation for liver failure.

Clinicopathologic features of atypical spindle cell lipomatous tumor / 中华病理学杂志

Objective@#To investigate the clinicopathologic characteristics, immunophenotype, differential and diagnostic features of atypical spindle cell lipomatous tumor (ASLT).@*Methods@#Three cases of ASLT were collected from January 2010 to March 2017 at Zhejiang Provincial People′s Hospital. The clinical and imaging features, histomorphology, immunophenotype and prognosis were analyzed. Fluorescence in situ hybridization (FISH) was used to detect MDM2 gene amplification, and relevant literature was reviewed.@*Results@#All three patients were adult males, aged 38, 43 and 54 years, respectively. One tumor originated in the subcutaneous soft tissue in the head and neck, one was located in the left primary bronchus and one in the latissimus dorsi muscle. Grossly, all three tumors were circumscribed and ranged from 4.0 to 5.8 cm in size. Microscopically, all showed a focally infiltrative front. These tumors were composed of variable proportions of spindle-shaped and adipocytic cells in a background of variable fibrous and edematous matrix. Scattered lipoblasts were easily seen. One tumor was composed predominately of spindle tumor cells, one of adipocytic cells, and one of equally mixed cell populations. The spindle tumor cells were generally bland-appearing with focal nuclear enlargement and hyperchromasia noted in one case. Mitosis was not seen in neither the spindle cells nor the adipocytic cells. By immunohistochemistry, diffuse and strong reactivity to CD34 of the spindle cells was noted in all cases, definite loss of Rb expression was noted in one of three cases, and S-100 protein was expressed only in the adipocytic cells. INI-1 was intact and Ki-67 index was 1% to 3%. All other markers including CDK4, MDM2, STAT6, SOX10, CD99, bcl-2, β-catenin, CD117, GFAP, CK, EMA, SMA and desmin were negative. FISH of MDM2 was done in two cases, and both showed no amplification. The ASLT in the head and neck had two recurrences during 17 months of follow-up, whereas the tumor in the latissimus dorsi was free of disease during 33 months of follow-up.@*Conclusions@#ASLT is a rare subtype of low-grade adipocytic neoplasm and is distinctive from atypical lipomatous tumor/well-differentiated liposarcoma. The histomorpholgy of ASLT has significant heterogeneity and forms a continuous spectrum. ASLT needs to be distinguished from a series of benign and malignant soft tissue tumors.

Evaluación de umbrales mínimos de celularidad en unidades de sangre de cordon umbilical en el Instituto Nacional Materno Perinatal del Perú / Evaluation of minimum thresholds of cellularity in units of umbilical cord blood at Instituto Nacional Materno Perinatal in Peru

RESUMEN Con el objetivo de evaluar criterios mínimos de celularidad de las unidades de sangre de cordón umbilical (USCU) según los estándares NETCORD en el Instituto Nacional Materno Perinatal de Lima, Perú, se realizó un estudio transversal que incluyó 100 USCU; se determinó el volumen, el recuento de células nucleadas totales (CNT) por hematología y el número de células CD34+ totales, así como también la viabilidad celular, por citometría de flujo. Se encontró que el 56% de las USCU no cumplen los umbrales mínimos de celularidad para ser criopreservadas en un banco de sangre de cordón umbilical. Se encontró, además, que las USCU de recién nacidos de mayor peso y de sexo femenino presentan mayor volumen y recuentos de células. En conclusión, es necesario considerar estas variables para optimizar la colecta de las USCU y obtener mayores recuentos de células que permita almacenar unidades de alta calidad en un futuro banco de sangre de cordón umbilical en Perú.

ABSTRACT A cross-sectional study that included 100 units of umbilical cord blood (UCB) was conducted to evaluate the minimum criteria of cellularity in UCB units, according to NetCord standards at Instituto Nacional Materno Perinatal in Lima, Peru. The volume, total count of nucleated cells by hematological tests and total number of CD34+ as well as cell viability by flow cytometry were determined. The study revealed that 56% of UCB units do not fulfill the minimum criteria of cellularity to be cryopreserved in an umbilical cord blood bank. Furthermore, the UCB units of newborns who weighed more and were female had a higher volume and cell count. In conclusion, these variables must undoubtedly be considered to optimize the collection of UCB units and obtain greater cell counts that enable the storage of high-quality units in a future umbilical cord blood bank in Peru.

Clinical significance of CD40, Ki-67 and CD34 expressions in gastric cancer and adjacent tissues / 中国医师杂志

Objective To investigate the relationship between CD40,Ki-67,and CD34 expressions and clinical significance of gastric cancer.Methods The expressions of CD40,Ki-67,and CD34 were detected by immunohistochemistry in 80 cases of gastric carcinomas and adjacent mucosas.Results The high expression of CD40 was detected in 36.3％ (29/80) cases of gastric cancer tissues,and Ki-67 was 53.8％ (43/80).The high expression of CD40 was related to lymph node metastasis,depth of invasion,and the patient's prognosis.The high expression of Ki-67 was related to histological differentiation,depth of invasion,vascular invasion,and lymph node metastasis (P ＜ 0.05).Expression of CD40 was positively related to Ki-67 and CD34 in gastric carcinomas (P ＜ 0.05).Conclusions The high expressions of CD40,Ki-67,and CD34 in gastric cancers are related to the tumor proliferation,tumor angiogenesis,and the prognosis of patients.

Correlación entre características físicas neonatales y maternas con el recuento total de células nucleadas y de células cd34+ por microlitro en sangre de cordón umbilical / Correlation between neonatal and maternal physical characteristics with total nucleated cell count and cell count per microliter cd34+ in cord blood / Correlação entre características físicas neonatais e maternas com a contagem total de células nucleadas e células cd34 + por microlitro no sangue do cordão umbilical

Introducción: La sangre de cordón umbilical (SCU) ha sido reconocida como una fuente de células madre hematopoyéticas. Múltiples estudios han sido realizados con el propósito de determinar variables maternas y neonatales que afecten el volumen, el recuento total de células nucleadas y de células CD34+. El presente estudio correlaciona variables maternas y neonatales con el recuento total de células nucleadas y de CD34+ medidas en μL (microlitro). Materiales y Métodos: Estudio correlacional en el que se analizaron 50 muestras de sangre de cordón umbilical de gestantes atendidas en dos IPS, una de Bogotá y otra de Ubaté, (Cundinamarca), durante un período de 7 meses del año 2013. La selección de estas muestras fue basada en un muestreo no probabilístico. Se calculó el coeficiente de correlación de Pearson con su respectiva significancia estadística entre las variables cuantitativas de la madre y del neonato, y el volumen, el recuento de células CD34+ y el recuento total de células nucleadas por μL. Resultados: Se encontró correlación positiva entre la longitud del cordón y el recuento total de células nucleadas. Así mismo entre el volumen inicial de la muestra y el recuento total de células nucleadas y el recuento de células CD34+ en μL y en mL (mililitro). Discusión: Múltiples estudios se han desarrollado entorno a la búsqueda de variables maternas y neonatales que afectan la calidad de la muestra. En Colombia no se habían descrito las correlaciones encontradas en el presente estudio. Es por ello, que el desarrollo de nuevos estudios con diseños analíticos será enriquecedor.

Background: The cord blood has been recognized as a source of hematopoietic stem. Multiple studies have been conducted in order to determine maternal and neonatal variables that affect the volume, total nucleated cell count and CD34+ cells. This study correlated maternal and neonatal variables with the total nucleated cell count and CD34+ cells measured by μL. Materials and methods: Correlational study in which 50 samples of umbilical cord blood were analyzed of pregnant women at two health institutions, one at Bogotá and the other one at Ubaté, Cundinamarca, for a period of seven months of 2013. The selection of these samples was based on a non-probability sample. The Pearson correlation coefficient was calculated with their respective statistical significance between quantitative variables of mother and newborn, and the volume, the count of CD34+ cells and the total nucleated cell count measured by μL. Results: Positive correlation between the length of the umbilical cord and the total nucleated cell count was found. Similarly, correlation between total nucleated cell count with the initial volume and count of CD34+ cells in μL and mL was found. Discussion: Several studies have been conducted around to look maternal and neonatal variables that affect the sample quality, however, the Colombian literature has not described the correlation found in this study, so it requires the development of new research with designs of analytical studies to establish associations between the variables described.

Introdução: O sangue do cordão umbilical (SCU) tem sido reconhecido como uma fonte de células-tronco hematopoiéticas. Vários estudos têm sido realizados com a finalidade de determinar as variáveis maternas e neonatais que afetam o volume e a contagem total de células nucleadas e células CD34+. O presente estudo correlaciona as variáveis maternas e neonatais com a contagem total das células nucleadas e células CD34+ medidas em μL (microlitro). Materiais e métodos: Estudo correlacional no que 50 amostras de sangue do cordão umbilical de mulheres grávidas atendidas em dois IPs foram analisadas, uma em Bogota e outra em Ubaté (Cundinamarca), durante um período de 7 meses do ano 2013. A seleção destas amostras foi baseada em uma amostragem não probabilística. O coeficiente de correlação de Pearson foi calculado com a respectiva significância estatística entre variáveis quantitativas da mãe e do recém-nascido, assim como o volume, a contagem de células CD34 + e contagem total de células nucleadas por μL. Resultados: Correlação positiva foi encontrada entre o comprimento do cordão e a contagem total de células nucleadas. Também entre o volume da amostra inicial e contagem total de células nucleadas e contagem de células CD34+ em μL e em mL (mililitro). Discussão: Vários estudos têm sido desenvolvidos em torno da busca de variáveis maternas e neonatais que afetam a qualidade da amostra. Na Colômbia não tinham sido descritas as correlações encontradas no presente estudo. É por esta razão que será gratificante o desenvolvimento de novos estudos com desenhos analíticos.

Study of novel coating strategy for coronary stents: simutaneous coating of VEGF and anti- CD34 antibody / Estudo da nova estratégia de revestimento para stents coronários: revestimento simultâneo de VEGF e anticorpo anti-CD34

Abstract Introduction: Intravascular coronary stenting has been used in the treatment of coronary artery disease (CAD), with a major limitation of in-stent restenosis (ISR). The 316 stainless steel has been widely used for coronary stents. In this study, we developed a novel coating method to reduce ISR by simultaneously coating vascular endothelial growth factor (VEGF) and anti-CD34 antibody on 316L stainless steel. Methods: Round 316L stainless steel sheets in the D-H group were polymerized with compounds generated from condensation reaction of dopamine and heparin using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS). Sixteen sheets from the D-H group were further immersed into 1ug/ml VEGF165 and 3mg/ml heparin sodium one after another for 10 times, and named as the D-(H-V)10 group. Eight sheets from the D-(H-V)10 group were coated with anti-CD34 antibody and termed as the D-(H-V)10-A group. Immunofluorescence assay and ELISA were used to evaluate whether the 316L stainless steel disks were successfully coated with VEGF and anti-CD34 antibody. Results: The results of immunofluorescence assay and ELISA showed that VEGF could be detected in the D-(H-V)10 and D-(H-V)10-A group, suggesting the steel sheets were successfully covered with VEGF. Anti-CD34 antibody could only be observed in the D-(H-V)10-A group, which was the only group coated with CD34 antibody. Both results suggested that the 316L stainless steel sheets were successfully coated with VEGF and anti-CD34 antibody. Conclusion: Our study developed a method to simultaneously coat VEGF and anti-CD34 antibody to stainless metal steel. This research serves as a fundamental role for a novel coating strategy. .

Resumo Introdução: O stent coronário intravascular tem sido utilizado no tratamento de doença arterial coronária, com uma maior limitação de restenose intra-stent (RIS). O aço inoxidável 316 tem sido amplamente utilizado para stents. Neste estudo, foi desenvolvido um novo método de revestimento para reduzir a RIS para revestir simultaneamente o fator de crescimento endotelial vascular (VEGF) e anti-CD34 em aço inoxidável 316L. Métodos: Placas de aço inoxidável 316L redondas no grupo DH foram polimerizadas com compostos gerados a partir da reacção de condensação de dopamina e heparina utilizando N- (3-dimetilaminopropil) -N'-etilcarbodiimida (EDC) e N-hidroxissuccinimida (NHS). Dezesseis folhas a partir do grupo DH foram ainda imersas em 1 ug/ml de VEGF 165 e 3 mg/ml de heparina sódica, um após outro por 10 vezes, sendo denominado como o grupo D-(HV)10. Oito folhas de D-(HV)10 foram revestidas com anticorpo anti-CD34 e denominado como grupo D-(HV)10-A. Testes de imunofluorescência e ELISA foram usados para avaliar se os discos de aço inoxidável 316L foram revestidos com sucesso com VEGF e anticorpo anti-CD34. Resultados: Os resultados dos testes de imunofluorescência e ELISA mostraram que o VEGF pôde ser detectado nos grupos D-(HV)10 e D-(HV)10-A, evidenciando que as chapas de aço foram cobertas com VEGF com sucesso. O anticorpo anti-CD34 podia apenas ser observado no grupo D-(HV)10-A, o único grupo revestido com anticorpo CD34. Ambos os resultados sugerem que as chapas de aço inoxidável 316L foram revestidas com sucesso com VEGF e anticorpo anti-CD34. Conclusão: Nosso estudo desenvolveu um método para revestir simultaneamente VEGF e anti-CD34 de aço inoxidável. Esta pesquisa tem um papel fundamental para a nova estratégia de revestimento. .

Application of D2-40/S100 and CD34/S100 detection in the diagnosis of blood and lymphatic vessel invasion of cutaneous malignant melanoma / 中华皮肤科杂志

Objective To develop an immunohistochemical assay for the diagnosis of cutaneous malignant melanoma (CMM) micrometastasis via blood and lymphatic vessels,and to evaluate its clinical significance.Methods Fifty-three patients (32 males and 21 females) histopathologically diagnosed as CMM were enrolled in this study.The patients were aged (61.2 ± 8.4) years (range,52-72 years).Tissue specimens were obtained from the central area of tumor in each case,and also from removed lymph nodes in some cases.The average duration of follow-up was (65.00 ± 5.68) months.During the follow-up,17 patients died of the recurrence or metastasis of CMM,and 6 patients were lost to follow-up.The expressions of D2-40,S100 and CD34 antigens in 53 tissue specimens were examined by immunohistochemical staining with three individual monoclonal antibodies,or by an immunohistochemical method using 2 two-antibody cocktails (D2-40/S 100 and CD34/S100) and double-color chromogens in single tissue sections.Results Of the 53 patients,30.19％ (16/53) were positive for hematoxylin-eosin (HE) staining combined with immunohistochemical staining with individual monoclonal antibodies,and 49.06％ (26/53) for the immunohistochemical method using two-antibody cocktails and double-color chromogens.Statistical differences were found in the positive rate between the two methods (x2 =3.94,P＜ 0.05).Compared with patients without blood/lymph vessel tumor emboli,those with blood/lymph vessel tumor emboli showed higher lymph node metastasis rate (80.77％ (21/26) vs.37.04％ (10/27),x2 =10.43,P ＜ 0.001),but lower five-year survival rate (42.31％ (11/26) vs.70.37％ (19/27),x2 =4.25,P ＜ 0.05).Conclusions The immunohistochemical method with two-antibody cocktails is superior to HE staining combined with immunohistochemical staining with individual monoclonal antibodies in the detection of blood/lymph vessel tumor emboli.And blood/lymph vessel tumor emboli may be an important prognostic factor in patients with CMM.

Increased angiogenesis in primary myelofibrosis: latent transforming growth factor-ß as a possible angiogenic factor.

OBJECTIVE: The aim of this work was to demonstrate a possible relationship between anti-latency-associated peptide human latent transforming growth factor beta 1 (latent TGF-ß1) expression in megakaryocytes and microvascular density in bone marrow biopsies from patients with essential thrombocythemia and primary myelofibrosis. METHODS: Microvascular density was evaluated by immunohistochemical analysis and the expression of latent TGF-ß1 in samples (100 megakaryocytes per bone marrow sample) from 18 essential thrombocythemia and 38 primary myelofibrosis (19 prefibrotic and 19 fibrotic) patients. Six bone marrow donor biopsies were used as controls. Fibrosis in the bone marrow biopsies was evaluated according to the European Consensus. RESULTS: The average fibrosis grade differed between essential thrombocythemia and primary myelofibrosis groups when compared to the control group. Latent TGF-ß1 expression differed significantly between the fibrotic primary myelofibrosis (PMF) group and the control group (p-value<0.01). A high degree of neo-angiogenesis (demonstrated by analysis of CD34 expression) was detected in patients with myelofibrosis. There were correlations between latent TGF-ß1 expression and microvascular density (r=0.45; p-value<0.0009) and between degree of microvascular density and fibrosis grade (r=0.80; p-value<0.0001). Remarkable differences for neo-angiogenesis were not observed between patients with essential thrombocythemia and controls. CONCLUSION: Angiogenesis participates in the pathogenesis of primary myelofibrosis, in both the prefibrotic and fibrotic stages, while latent TGF-ß is differentially expressed only in the prefibrotic stage.

Increased angiogenesis in primary myelofibrosis: latent transforming growth factor-β as a possible angiogenic factor

Objective: The aim of this work was to demonstrate a possible relationship between anti-latency-associated peptide human latent transforming growth factor beta 1 (latent TGF-β1) expression in megakaryocytes and microvascular density in bone marrow biopsies from patients with essential thrombocythemia and primary myelofibrosis. Methods: Microvascular density was evaluated by immunohistochemical analysis and the expression of latent TGF-β1 in samples (100 megakaryocytes per bone marrow sample) from 18 essential thrombocythemia and 38 primary myelofibrosis (19 prefibrotic and 19 fibrotic) patients. Six bone marrow donor biopsies were used as controls. Fibrosis in the bone marrow biopsies was evaluated according to the European Consensus. Results: The average fibrosis grade differed between essential thrombocythemia and primary myelofibrosis groups when compared to the control group. Latent TGF-β1 expression differed significantly between the fibrotic primary myelofibrosis (PMF) group and the control group (p-value < 0.01). A high degree of neo-angiogenesis (demonstrated by analysis of CD34 expression) was detected in patients with myelofibrosis. There were correlations between latent TGF-β1 expression and microvascular density (r = 0.45; p-value < 0.0009) and between degree of microvascular density and fibrosis grade (r = 0.80; p-value < 0.0001). Remarkable differences for neo-angiogenesis were not observed between patients with essential thrombocythemia and controls. Conclusion: Angiogenesis participates in the pathogenesis of primary myelofibrosis, in both the prefibrotic and fibrotic stages, while latent TGF-β is differentially expressed only in the prefibrotic stage...

Avaliação da angiogênese em lesões de líquen plano oral e pênfigo vulgar / Assessment of angiogenesis in lesions of oral lichen planus and pemphigus vulgaris

O Líquen Plano Oral e o Pênfigo Vulgar são doenças imunológicas crônicas mucocutâneas de etiologias desconhecidas que podem acometer a mucosa oral. Tão importante quanto em neoplasias o papel da angiogênese na patogenia das doenças inflamatórias crônicas é de apreciável interesse, podendo ser considerado um marcador da atividade da doença. Ademais, com o estudo do processo angiogênico nessas lesões tem-se a possibilidade de aprimorar o entendimento do mecanismo patogênico nas mesmas. Este trabalho se propôs a avaliar a atividade angiogênica através da expressão imunoistoquímica do anticorpo anti-CD34, utilizando a técnica da MVC, em 26 casos de LPO reticulares, 14 casos de LPO erosivos, 18 casos de PV e em 15 casos de mucosa oral normal. Os resultados deste experimento foram submetidos a testes estatístico não-paramétricos com nível de significância de 5%. Não foram observadas diferenças estatisticamente significativas entre o número médio de vasos obtidos pela MVC nas lesões. Todavia, foi observado que a mediana dos casos de líquen plano oral foi maior que no grupo de pênfigo vulgar, que por sua vez revelou mediana maior que nos casos de mucosa oral normal (p=0,280). No tocante a contagem microvascular de CD34 em relação à forma clínica dos líquens planos orais (reticular e erosivo), verificou-se uma mediana ligeiramente maior para a forma clínica erosiva. Apesar de os testes estatísticos utilizados não demonstrarem diferença significativa (p=0,720). Ainda que os resultados do presente estudo não permitam a afirmação contundente da participação do processo angiogênico na patogênese e progressão das lesões de líquen plano oral e pênfigo vulgar, sugerimos que este processo está presente em ambas as lesões, porém outros estudos devem ser realizados a fim de que essa hipótese seja fundamentada. (AU)

Oral lichen planus and pemphigus vulgaris are chronic diseases mucous membrane immune of unknown aetiology that can be observed affecting to the oral mucous. A relevant as regards neoplasies the role angiogenesis in the inflammatory chronic disease pathogenesis as it provides a substancial interest can be considered as being an activity diseases marker; besides being through specialised research of this angiogenic process to improve of understanding pathogenic mechanism. This research proposes to assess angiogenic active through of antibody immunohistochemistry expression antiCD34 antibody in 26 OLP of reticular cases, 14 OLP erosives cases, 18 of PV cases and 15 specimens of normal oral mucosa. The result was submitted non-parametric tests of 5% significance level. It is not statistically significant correlacion was seen regarding between average vessels. However, only be effectively observed the median of OLP cases was larger than pemphigus vulgaris in fact proved average larger than oral normal mucosa (p=0,280). Regarding the microvascular count of CD34 concerning clinic form oral lichen planus (reticular and erosion) increased emphasis is more cross-border average for the form erosion clinic. Despite of the statistic tests could not be more effective (p=0,720). Even though, the results of the research is not sufficient to enable to consider of angiogenic process in the pathogenesis and lesions progression of oral lichen planus and pemphigus vulgaris, we suggest this process is present in both forms lesion, however, more studies must be made in the near future in order to prepare a well-founded proposal. (AU)

Preliminary recognition of stem cells surface markers in hepatoblastoma / 重庆医学

Objective To preliminarily screen the neoplastic stem cell (NSC) related surface markers combination and to under-stand the NSC distribution in hepatoblastoma(HB) .Methods The children cases of HB undergone the surgical therapy in the Jian-gxi Provincial Children′s Hospital were selected .The immunohistochemical method was adopted for observing the expression and distribution of NSC-related markers CD34 ,Thy-1 ,c-kit ,CD56 and stem cell factor(SCF) in the HB tissue and the normal hepatic tissue away from the edge of tumor tissue 3cm outside .Results Thy-1 and c-kit were sporadically distributed in the HB tissue and mainly focued on the portal area ,but did not exressed in the normal liver tissue ;the expression of CD34 and SCF in HB was signifi-cantly higher than that in the normal liver tissue(P0 .05) .Conclusion The different NSC related surface markers are distributed in the HB tissue and focus on the specific areas .The positive cells of Thy-1/c-kit expression may play a role in the HB occurrence .

Análise da imunoexpressão da podoplanina (D2-40), da proteína nuclear Ki-67 e da catepsina D-34 e suas relações com metástase em pacientes operados por tumor carcinoide típico broncopulmonar / Analysis of the immunoexpression of podoplanin (D2-40), of the nuclear protein Ki-67, and of cathepsin D-34 and their relation to metastasis in patients operated for typical bronchopulmonary carcinoid tumor

Os tumores carcinoides broncopulmonares típicos são considerados tumores de baixo grau de malignidade, o que faz com que muitas vezes seja negligenciada sua capacidade de gerar metástases e levar a óbito. A atual impossibilidade de se fazer predições fidedignas do potencial metastático para individualizar a terapêutica e o manejo clínico do paciente portador de tumor carcinoide típico muitas vezes impõe situações de dúvida nas decisões da prática clínica diária. O conhecimento das características moleculares e genéticas desses tumores é a esperança de melhor compreensão de sua história natural, da identificação de seus fatores prognósticos e da avaliação de novas propostas terapêuticas. O objetivo deste estudo foi quantificar três imunomarcadores: o D2-40, o CD-34 e o Ki-67 e avaliar se eles são capazes de predizer metástase. Como informação adicional também foram avaliadas as variáveis clínicas e suas associações com metástases. Material e métodos: Foram analisados prontuários de 97 pacientes submetidos à cirurgia para tratamento de tumor carcinoide típico broncopulmonar e que apresentavam período de acompanhamento pós operatório de cinco anos completos. Foi estabelecido um banco de dados epidemiológicos, anátomopatológicos, cirúrgicos e clínicos relacionados à doença. Os blocos de parafina contendo os tumores primários, metastáticos e os linfonodos ressecados foram recuperados e reavaliados por dois patologistas para confirmação do diagnóstico histológico. Após confirmação diagnóstica foram realizadas as reações imunohistoquímicas para o D2-40, o CD34 e Ki-67. As variáveis demografia, gênero, idade, localização do tumor, tamanho do tumor, margem comprometida, total de linfonodos dissecados e a quantificação dos marcadores Ki-67, CD34 e podoplanina (área linfática e densidade) foram comparadas a cinco desfechos: metástase linfática; metástase em linfonodos hilares, peribrônquicos e pulmonares ipsilaterais (N1); metástase em linfonodos mediastinais...

Typical bronchopulmonary carcinoid tumors are slow-growing tumors considered to be of low malignancy, a fact that often underscores their capacity to generate metastasis that leads to death. The literature does not contain any assessment of the metastatic potential that would allow for individualized and optimized therapy that could have a positive impact upon the survival rate of patients. Genetic and biomolecular studies are the most promising venues for better comprehension of the behavior and natural history of such tumors. The objective of this investigation was to analyze three immunomarkers associated to malignity phenotypes: D2-40, CD-34, and Ki-67, and to verify whether or not they are associated to metastasis. D2-40 is a specific marker of the proliferation of the lymphatic endothelium and allows for the quantification of lymphangiogenesis. CD-34 identifies the endothelial cells of micro vessels and quantifies angiogenesis. Ki-67 in turn is a marker of neoplasia cell proliferation. As additional information several clinical variables were scrutinized for their association to metastasis. Ninety-seven subjects were assessed herein. These had undergone surgery for typical bronchopulmonary carcinoid tumor and all had complied with a full 5-year follow-up period. The paraffin blocks containing the primary and metastatic tumors and the dried lymph nodes were recovered. Once the histologic diagnosis was confirmed immunohistochemical reactions were performed for D2-40, Ki-67, and CD-34. The variables demography, gender, age, tumor site, tumor size, compromised margin, total dissected lymph nodes and quantification of D2-40 (lymphatic area and density), Ki-67 and CD-34 markers were compared to 5 outcomes: lymphatic metastasis, metastasis in hilar, peribronchic and ipsilateral pulmonary lymph nodes (N1), metastasis in ipsilateral mediastinal lymph nodes (N2), haematogenic metastasis, and lymphatic or haematogenic metastasis. It was concluded that there is...

Chimerism of placenta-derived cells with maternal blood and umbilical cord blood cells / 中国组织工程研究

BACKGROUND:There are abundant cel populations in the placenta that attracts more and more attentions because of high content of CD34+cel s. It is expected to become a new source of hematopoietic stem cel s for the treatment of hematologic diseases and other malignant diseases. OBJECTIVE:To investigate the amount of cel s derived from placenta, their colony forming ability, and their chimerism analysis. METHODS:Five placentas obtained from five healthy ful-term cesarean women were treated with perfusion method and tissue digestion for the cel col ection. Flow cytometry was used to detect the proportion of CD34+cel s in the placenta and cord blood, fol owed by the culture of cel colonies as wel as regular observation of cel morphology and counting. PCR amplification with sequence-specific primers and sequence-specific oligonucleotide probes were used to examine HLA type of placenta, umbilical cord blood, and maternal peripheral blood;Short tandem repeat PCR was used for chimerism analysis. RESULTS AND CONCLUSION:There were more CD34+cel s in the placenta than in the umbilical cord blood. The placenta had good ability to form multiple colonies in vitro, and there were maternal source components in the placenta. It is concluded that the amount of cel s in the placenta and their biological functions exhibit the potential use of placenta as a new source of hematopoietic stem cel s.

Mononuclear cells isolated from mobilized bone marrow differentiate into vascular endothelial cells and cardiomyocyte-like cells / 中国组织工程研究

BACKGROUND:It is controversial whether bone marrow mobilization can retain in cardiac injured position in congestive cardiomyopathy or differentiate into cardiomyocytes and vascular endothelial cells. OBJECTIVE:To study the effects of granulocyte colony stimulating factor (G-GSF) on myocardium and angiogenesis in rats with congestive cardiomyopathy. METHODS:Fifty Wistar rats with heart failure caused by adriamycin-induced cardiomyopathy were divided into heart failure group (n=20) treated with normal saline and bone marrow mobilization (n=30) treated with subcutaneous injection of recombinant human G-GSF. Ten rats from the bone marrow mobilization were kil ed at day 6 of mobilization, and myocardial tissue was taken for CD43 immunofluorescent staining. Blood samples were taken from the rat tail in each group before and 5 days after treatment to count total number of white blood cells and percentage of mononuclear cells. Meanwhile, mononuclear cells extracted from the peripheral blood were used for flow cytometry detection. At day 5 after treatment, bromodeoxyuridine (BrdU, 50 mg/kg) was successively given to al rats for 4 weeks before they were sacrificed. Myocardial tissues were taken to determine the homing of mononuclear cells and evaluate differentiation of mononuclear cells into cardiomyocytes and vascular endothelial cells using BrdU staining, BrdU/myosin heavy chain double staining, and BrdU/actin double staining. Hematoxylin-eosin staining was used for determination of blood vessel density. RESULTS AND CONCLUSION:G-CSF mobilization increased the number of mononuclear cells that was significantly higher than before treatment (P<0.05). Flow cytometry showed that the number of CD34-positive mononuclear cells in the peripheral blood was higher in the bone marrow mobilization than in the heart failure group (P<0.05). Myocardial CD34 immunofluorescence showed that the heart failure group was negative and the bone marrow mobilization group was positive. In the bone marrow mobilization group, the myocardial tissue was positive for BrdU staining, BrdU/myosin heavy chain double staining and BrdU/actin double staining, while vascular endothelial cells in the region of myocardial injury was positive for BrdU;conversely, the heart failure group was negative. The density of blood vessels in the bone marrow mobilization group was significantly higher than that in the heart failure group (P<0.001). These findings indicate that bone marrow mobilization increases the number of mononuclear cells, and these cells are homing to myocardial injury, thereby playing a repair role in the myocardium and vascular tissue of heart failure rats with congestive cardiomyopathy.