Leiomiomatosis vulvovaginal: reporte de un caso y revisión bibliográfica / Vulvo-Vaginal Leiomyomatosis: Case Report and Review of the Literature

Resumen ANTECEDENTES: Los leiomiomas son los tumores pélvicos más frecuentes en la mujer; sin embargo, su localización vaginal es excepcional. Suelen ser asintomáticos y encontrarse como un hallazgo clínico. En los últimos 20 años solo se han reportado 85 casos, y solo dos han sido recurrentes. OBJETIVO: Reportar un caso de miomatosis vaginal recurrente en una paciente histerectomizada y revisar la bibliografía al respecto. CASO CLÍNICO: Paciente de 58 años, histerectomizada, con una tumoración vaginal. El reporte histopatológico informó una proliferación fusocelular, debidamente delimitada, dispuesta en haces entrecruzados. Los núcleos eran alargados, monomorfos y de extremos romos. El estroma era escaso y colagénico. No se observaron atipias citonucleares ni necrosis. El estudio inmunohistoquímico de la lesión con actina de anticuerpos antimúsculo liso y desmina se reportó positivo. Se diagnosticó miomatosis vulvovaginal recurrente. Se trató mediante resección quirúrgica. CONCLUSIÓN: Los leiomiomas vulvovaginales son extremadamente raros y la bibliografía al respecto es poca; su recurrencia es verdaderamente excepcional. De ahí la importancia de la publicación de estos casos, que aporta información que pueden tomar en cuenta otros clínicos al momento del diagnóstico.

Abstract BACKGROUND: Leiomyomas are the most frequent pelvic tumors in women; however, their vaginal location is unusual. They are usually asymptomatic and present as a clinical finding. In the last 20 years only 85 cases have been reported, and only two have been recurrent. OBJECTIVE: To report a case of recurrent vaginal myomatosis in a hysterectomized patient and review the literature. CLINICAL CASE: A 58-year-old hysterectomized patient with a vaginal tumor. The histopathologic report reported a fusocellular proliferation, properly delimited, arranged in crisscross bundles. The nuclei were elongated, monomorphous and blunt ended. The stroma was sparse and collagenous. No cytonuclear atypia or necrosis were observed. Immunohistochemical study of the lesion with anti-smooth muscle antibody actin and desmin was reported positive. Recurrent vulvovaginal myomatosis was diagnosed. It was treated by surgical resection. CONCLUSION: Vulvovaginal leiomyomas are extremely rare, and the literature is sparse; their recurrence is truly exceptional. Hence the importance of publishing these cases, providing information to be considered by other clinicians at the time of diagnosis.

Fascin overexpression in nasal mucosa in patients with chronic rhinosinusitis with and without nasal polyps / Superexpressão de fascina na mucosa nasal em pacientes com rinossinusite crônica com e sem pólipos nasais

ABSTRACT Objective: The aim of our research is to assess fascin expression in nasal tissues of patients with chronic rhinosinusitis with (CRSwNP) and without (CRSsNP) nasal polyps. Methods: Fascin expression in nasal tissues of 11 CRSwNP patients and 10 CRSsNP patients was immunohistochemically evaluated and compared with control subjects. Results: Fascin was found to be strongly expressed in epithelial cells in polyps in CRSwNP and nasal tissue in CRSsNP. Its strong expression was observed both in lamina propria and nasal epithelial cells in CRSsNP. Fascin overexpression in nasal mucosa in CRSwNP was more pronounced compared with CRSsNP. In addition, proliferating epithelial cells in polyp tissue were weakly immunostained, whereas mature cells expressed much more fascin. Conclusion: CRSwNP and CRSsNP are associated with fascin overexpression, which makes fascin a promising target for therapeutic interventions.

RESUMEN Objetivo: El objetivo de esta investigación fue evaluar la expresión de fascina en los tejidos nasales de pacientes con rinosinusitis crónica con (RSCcPN) y sin pólipos nasales (RSCsPN). Métodos: La expresión de fascina en los tejidos nasales de 11 pacientes con RSCcPN y 10 pacientes con RSCsPN fue analizada por inmunohistoquímica y comparada con los individuos control. Resultados: Fascina fue encontrada por ser fuertemente expresada en células epiteliales de pólipos en la RSCcPN y en tejido nasal en la RSCsPN. Su fuerte expresión fue observada tanto en la lámina propia como en las células epiteliales nasales en la RSCsPN. La sobreexpresión de fascina en la mucosa nasal en la RSCcPN fue más pronunciada en comparación con la RSCsPN. Además, las células epiteliales proliferantes en el tejido del pólipo fueron inmunoteñidas débilmente, mientras las células maduras expresaron mucho más fascina. Conclusión: RSCcPN y RSCsPN están asociadas a la sobreexpresión de fascina, lo que hace la fascina un objetivo prometedor para intervenciones terapéuticas.

RESUMO Objetivo: O objetivo desta pesquisa foi avaliar a expressão de fascina nos tecidos nasais de pacientes com rinossinusite crônica com (RSCcPN) e sem (RSCsPN) pólipos nasais. Métodos: A expressão de fascina nos tecidos nasais de 11 pacientes com RSCcPN e 10 pacientes com RSCsPN foi avaliada imuno-histoquimicamente e comparada com os indivíduos-controle. Resultados: Fascina foi encontrada por ser fortemente expressa em células epiteliais em pólipos na RSCcPN e em tecido nasal na RSCsPN. Sua forte expressão foi observada tanto na lâmina própria quanto nas células epiteliais nasais na RSCsPN. A superexpressão de fascina na mucosa nasal na RSCcPN foi mais pronunciada em comparação com a RSCsPN. Além disso, as células epiteliais em proliferação no tecido do pólipo foram imunocoradas fracamente, enquanto as células maduras expressaram muito mais fascina. Conclusão: RSCcPN e RSCsPN estão associadas à superexpressão de fascina, o que torna a fascina um alvo promissor para intervenções terapêuticas.

Dact1 involvement in the cytoskeletal arrangement of cardiomyocytes in atrial fibrillation by regulating cx43

Abstract Objective: To determine the role of the dishevelled binding antagonist of beta catenin 1 (DACT1) in the cytoskeletal arrangement of cardiomyocytes in atrial fibrillation (AF). Methods: The DACT1 expression and its associations with the degree of fibrosis and β-catenin in valvular disease patients were analyzed by immunohistochemistry and Masson's staining. DACT1 was overexpressed in the atrial myocyte cell line (HL-1) and the cardiac cell line (H9C2) by adenoviral vectors. Alterations in the fibrous actin (F-actin) content and organization and the expression of β-catenin were detected by flow cytometry, immunofluorescence, and Western blotting. Additionally, the association of DACT1 with gap junctions connexin 43 (Cx43) was detected by immunohistochemistry, immunofluorescence, and Western blotting. Results: Decreased cytoplasmic DACT1 expression in the myocardium was associated with AF (P=0.037) and a high degree of fibrosis (weak vs. strong, P=0.028; weak vs. very strong, P=0.029). A positive association was observed between DACT1 and β-catenin expression in clinical samples (P=0.028, Spearman's rho=0.408). Furthermore, overexpression of DACT1 in HL-1 and H9C2 cells induced an increase in β-catenin and subsequent partial colocalization of DACT1 and β-catenin. In addition, F-actin content and organization were enhanced. Interestingly, DACT1 was positively correlated with the Cx43 expression in clinical samples (P=0.048, Spearman's rho=0.370) and changed the Cx43 distribution in cardiac cell lines. Conclusion: DACT1 proved to be a novel AF-related gene by regulating Cx43 via cytoskeletal organization induced by β-catenin accumulation in cardiomyocytes. DACT1 could thus serve as a potential therapeutic marker for AF.

La leptina promueve la expresión de Hic-5 y la formación de puntos de actina por la vía dependiente de FAK-Src en células epiteliales mamarias MCF10A / Leptin induced Hic-5 expression and actin puncta formation by the FAK/Src-dependent pathway in MCF10A mammary epithelial cells

Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

Expression of microtubule actin cross-linking factor 1 in gastric carcinoma and its effect on cell invasion and metastasis / 中华消化杂志

Objective@#To explore the role of microtubule actin cross-linking factor 1(MACF1) in the metastasis of gastric cancer.@*Methods@#From 2009 to 2012, at The First Affiliated Hospital of Zhengzhou University, the paraffin blocks of gastric cancer and normal tissue adjacent to cancer of 107 patients who received radical gastrectomy were collected. The expression of MACF1 in tissues at protein level was detected by immunohistochemical staining. In 2017, at The First Affiliated Hospital of Zhengzhou University, fresh specimens samples of gastric cancer and normal tissue adjacent to cancer of 42 patients who received radical gastrectomy were also collected. The expression of MACF1 at mRNA level was determined by quantitative real-time polymerase chain reaction (PCR). MACF1 knockout gastric cell line was established. The effects of MACF1 on cell migration and invasion were verified by wound-healing test and Transwell assay. The effects of MACF1 on cell microtubule and actin were analyzed by filamentous actin (F-actin) staining. T-test, chi-square test and multivariate analysis were used for statistical analysis.@*Results@#The positive expression rate of MACF1 in gastric carcinoma tissues was 71.0%(76/107), which was significantly higher than that of the corresponding normal tissues adjacent to cancer (22.4%, 24/107), and the difference was significant (t=4.145, P=0.016). The expression of MACF1 at mRNA level in cancer tissues of 42 patients with gastric cancer was 6.463±0.672, which was significantly higher than that of corresponding normal tissue adjacent to cancer (1.727±0.331), and the difference was statistically significant (t=6.326, P<0.01). The differences in positive expression rate of MACF1 in different tumor infiltration depth, different TNM stage and lymph nodes metastasis were statistically significant (χ2=1.170, 7.959 and 5.288; all P<0.01). The five-year survival rate of patients with high expression of MACF1 was 32.9% (25/76), which was significantly lower than that of patients with normal MACF1 expression (64.5%, 20/31), and the difference was statistically significant (χ2=25.093, P=0.034). The high expression of MACF1 was an independent prognostic factor affecting overall survival rate in patients with gastric cancer after surgery(hazard ratio (HR)=0.513, 95%confidence interval (CI): 0.411 to 0.922, P=0.038). The results of wound-healing assay showed that at 24 hour after wound the migration ability of MACF1 knockout AGS- MACF1-/- cells was (18.77±3.82)%, which was lower than that of wild type AGS cells ((76.24±5.36)%), and the difference was statistically significant (t=6.249, P=0.014). The migration ability of MACF1 knockout HGC27-MACF1-/-cells was (42.48±7.37)%, which was lower than that of wild type HGC27 cells ((82.35±4.28)%), and the difference was statistically significant (t=5.938, P=0.017). The results of Transwell assay indicated that the number of migration cells of MACF1 knockout AGS-MACF1-/- cells was 87.0±11.0, which was less than that of wild type AGS cells (200.0±16.0), and the difference was statistically significant (t=5.820, P=0.028). The number of migration cells of MACF1 knockout HGC27-MACF1-/-cells was 151.0±13.0, which was less than that of wild type HGC27 cells (268.5±20.5), and the difference was statistically significant (t=4.840, P=0.040). The results of F-actin staining demonstrated that the number of actin filaments of MACF1 knockout AGS-MACF1-/- cells was 216.60±18.09, which was less than that of wild type AGS cells (491.30±5.02), and the difference was statistically significant (t=14.630, P=0.005).@*Conclusions@#The abnormally high expression of MACF1 in gastric cancer tissues may be correlated with the poor prognosis of patients with gastric cancer. MACF1 promotes the invasion and metastasis of gastric cancer cells by affecting the formation of F-actin and cell skeleton.

The esophageal expression of fibrotic cytokines and NADPH oxidase in stress induced esophageal fibrosis and its significance / 中国医师杂志

Objective To detect the expression levels of collagen1 (colla-1),transforming growth factor-β1 (TGF-β1),a-smooth muscle actin (α-SMA) and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX-4) in mouse esophagus submitted to chronic restraint stress (CRS),in order to discuss stress-induced esophageal fibrosis and the role of oxidative stress.Methods 20 male Kunming mice were randomly divided into two groups,CRS and normal control (NC).The mice in CRS group were submitted to 2 h per day of restraint stress using home-made device for a period of 14 days,and the mice in both group were treated the same at rest of the time.Fibrotic changes of esophageal tissue were observed using Masson staining.The expression levels of NOX-4 and related fibrotic cytokines in esophageal tissues were detected by several methods such as immunohistochemistry,enzyme-linked immunosorbent assay (ELISA) and realtime polymerase chain reaction (qRT-PCR).Results Body weight in CRS group was significantly lower than NC group (8.75 ± 1.69 vs 12.69 ± 3.16),with statistically significant difference (t =3.11,P ＜ 0.05).Masson staining revealed that CRS mice showed distinct fibrosis of epithelial interstitium,while there was no distinct changes observed in NC mice.Immunohistochemical staining revealed intense staining for NOX-4 in epithelial,mucosal and submucosal layers of esophagi in CRS mice.ELISA showed that the serum level of NOX-4 in CRS mice was higher than NC mice (1.442 ± 0.05 vs 0.449 ± 0.08),with statistically significant difference (t =-27.32,P ＜ 0.01).Real-time PCR results showed that the expression of colla-1,TGF-β1,α-SMA and NOX-4 in CRS mice were as (2.443 ±0.36,2.78 ±0.13,2.244 ±0.18,2.448 ±0.440) times higher than NC mice,with statistically significant difference (t =-11.19,-38.86,-19.90,-10.37,P ＜ 0.01).Conclusions Fibrotic cytokines such as colla-1,TGF-β1 and α-SMA may participate in formation of stress induced esophageal fibrosis,and oxidative stress may play crucial role in the process of esophageal fibrosis.

Expression of microtubule actin cross-linking factor 1 in gastric carcinoma and its effect on cell invasion and metastasis / 中华消化杂志

Objective To explore the role of microtubule actin cross-linking factor 1 (MACF1) in the metastasis of gastric cancer.Methods From 2009 to 2012,at The First Affiliated Hospital of Zhengzhou University,the paraffin blocks of gastric cancer and normal tissue adjacent to cancer of 107 patients who received radical gastrectomy were collected.The expression of MACF1 in tissues at protein level was detected by immunohistochemical staining.In 2017,at The First Affiliated Hospital of Zhengzhou University,fresh specimens samples of gastric cancer and normal tissue adjacent to cancer of 42 patients who received radical gastrectomy were also collected.The expression of MACF1 at mRNA level was determined by quantitative real-time polymerase chain reaction (PCR).MACF1 knockout gastric cell line was established.The effects of MACF1 on cell migration and invasion were verified by wound-healing test and Transwell assay.The effects of MACF1 on cell microtubule and actin were analyzed by filamentous actin (F-actin) staining.T-test,chi-square test and multivariate analysis were used for statistical analysis.Results The positive expression rate of MACF1 in gastric carcinoma tissues was 71.0％ (76/107),which was significantly higher than that of the corresponding normal tissues adjacent to cancer (22.4％,24/107),and the difference was significant (t =4.145,P =0.016).The expression of MACF1 at mRNA level in cancer tissues of 42 patients with gastric cancer was 6.463 ±0.672,which was significantly higher than that of corresponding normal tissue adjacent to cancer (1.727 ± 0.331),and the difference was statistically significant (t =6.326,P ＜ 0.01).The differences in positive expression rate of MACF1 in different tumor infiltration depth,different TNM stage and lymph nodes metastasis were statistically significant (x2 =1.170,7.959 and 5.288;all P ＜ 0.01).The five-year survival rate of patients with high expression of MACF1 was 32.9％ (25/76),which was significantly lower than that ofpatients with normal MACF1 expression (64.5％,20/31),and the difference was statistically significant (x2 =25.093,P =0.034).The high expression of MACF1 was an independent prognostic factor affecting overall survival rate in patients with gastric cancer after surgery (hazard ratio (HR) =0.513,95％ confidence interval (CI):0.411 to 0.922,P =0.038).The results of wound-healing assay showed that at 24 hour after wound the migration ability of MACF1 knockout AGS-MACF1 / cells was (18.77 ± 3.82) ％,which was lower than that of wild type AGS cells ((76.24 ± 5.36) ％),and the difference was statistically significant (t =6.249,P =0.014).The migration ability of MACF1 knockout HGC27-MACF1-/-cells was (42.48 ± 7.37)％,which was lower than that of wild type HGC27 cells ((82.35-± 4.28) ％),and the difference was statistically significant (t =5.938,P =0.017).The results of Transwell assay indicated that the number of migration cells of MACF1 knockout AGS-MACF1-/-cells was 87.0 ± 11.0,which was less than that of wild type AGS cells (200.0 ± 16.0),and the difference was statistically significant (t =5.820,P =0.028).The number of migration cells of MACF1 knockout HGC27-MACF1-/-cells was 151.0 ± 13.0,which was less than that of wild type HGC27 cells (268.5 ± 20.5),and the difference was statistically significant (t =4.840,P =0.040).The results of F-actin staining demonstrated that the number of actin filaments of MACF1 knockout AGS-MACF1-/-cells was 216.60 ± 18.09,which was less than that of wild type AGS cells (491.30 ± 5.02),and the difference was statistically significant (t =14.630,P =0.005).Conclusions The abnormally high expression of MACF1 in gastric cancer tissues may be correlated with the poor prognosis of patients with gastric cancer.MACF1 promotes the invasion and metastasis of gastric cancer cells by affecting the formation of F-actin and cell skeleton.

Changes in expression of Talin1 and F-actin during serum-induced proliferation of pulmonary artery smooth muscle cells of rats with hepatopulmonary syndrome / 中华麻醉学杂志

Objective To evaluate the changes in the expression of Talin1 and F-actin during ser-um-induced proliferation of pulmonary artery smooth muscle cells ( PASMCs) of rats with hepatopulmonary syndrome ( HPS) . Methods Twenty healthy male Sprague-Dawley rats, weighing 220-250 g, were used for producing HPS by chronic ligation of the common bile duct. Blood samples from the abdominal aorta were collected to prepare serum. Primarily cultured PASMCs obtained from rats were seed in 6- or 96-well plates and divided into 2 groups using a random number table method: control group ( group C) and HPS group ( group HPS) , with 24 wells in each group ( for 6-well plates) or with 30 wells in each group ( for 96-well plates) . In C and HPS groups, normal rat serum or HPS rat serum were added, respectively, with the final concentration of 5％. At 24, 48 and 72 h of incubation, the expression of Talin1, F-actin and G-actin was determined by Western blot, the F-actin∕G-actin ratio was calculated, and the proliferation of PASMCs was measured by 3H-TdR incorporation and CCK-8 assays. Results Compared with group C, the proliferation of PASMCs was significantly enhanced, the expression of Talin1 was up-regulated, and the F-actin∕G-actin ratio was increased in group HPS ( P<0. 05) . The proliferation of PASMCs was gradually en-hanced, the expression of Talin1 was gradually up-regulated, and the F-actin∕G-actin ratio was gradually increased with the prolonged incubation time in group HPS (P<0. 05). Conclusion The mechanism by which the HPS rat serum induces proliferation of PASMCs may be related to up-regulating the expression of Talin1 and F-actin.

Prognostic factors analysis of Ki-67, α-SMA expression in retroperitoneal leiomyosarcoma / 中华肿瘤杂志

Objective@#To investigate the expression and prognostic value of alpha smooth muscle actin(α-SMA) and Ki-67 in retroperitoneal leiomyosarcoma.@*Methods@#Fifty retroperitoneal leiomyosarcoma patients who underwent operation in Chinese People′s Liberation Army General Hospital from May 2002 to December 2015 were retrospectively analyzed. There were 14 males and 36 females form 21 to 79 and an average age of 48. Kaplan-Meier estimations and Cox regression analyses were performed.@*Results@#Of the 50 cases, 45 patients underwent complete resection, and others are not. The overall 1, 3, 5-year survival rates were 86.0%, 46.0% and 28.0%, respectively. Tumor size, extent of resection, pathological stage, and expression levels of Ki-67 and alpha smooth muscle actin (α-SMA) were closely related to the survival of retroperitoneal leiomyosarcoma patients (all P<0.05), respectively. Multivariate analysis showed that pathological grade and degree of surgical resection were independent risk factors in the prognosis of patients (P<0.05).@*Conclusion@#The high expression of α-SMA and Ki-67 are indicators of poor prognosis in retroperitoneal leiomyosarcoma, which can be used as a potential survival predictor in patients with retroperitoneal leiomyosarcoma.

Mechanism of cucurbitacin B underlying pressure overload-induced myocardial fibrosis / 中华老年心脑血管病杂志

Objective To study the mechanism of cucurbitacin B (CuB) underlying pressure over load-induced myocardial fibrosis.Methods Sixty C57 mice were divided into sham operation group,CuB treatment group,aorta ligation group,CuB+aorta ligation group (15 in each group).The animals received intragastric gavage (0.2 mg/kg · 2 d) one week after operation and a myocardial fibrosis model of mice was induced by aorta ligation 4 weeks after operation.Microvascular density (MVD) was detected with immunohistochemical staining.Expressions of α-SMA,CD31,CD34,vimentin and endothelial-mesenchymal transition (EndMT) were detected by Western blot with immunofluorescence staining.Results No significant difference was found in cardiac MVD,and expression level of α-SMA,vimentin,CD31 and CD34 between sham operation group and CuB group 4 weeks after operation (P＞0.05).The cardiac MVD and expression level of CD31 and CD34 were significantly lower while the expression level of α-SMA and vimentin was significantly higher in aorta ligation group than in sham operation group 4 weeks after operation (P＜0.05).The cardiac MVD and expression level of CD31 and CD34 were significantly higher while the expression level of α-SMA and vimentin was significantly lower in CuB+aorta ligation group than in aorta ligation group 4 weeks after operation (P＜0.05).Conclusion CuB can attenuate cardiac fibrosis by regulating EndMT.

Detection of F-actin autoantibodies in the serum of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the value of F-actin autoantibodies in the serum of patients with systemic lupus erythematosus (SLE),and to explore the relationships between F-actin autoantibodies and other clinical indicators.Methods ELISA was established to detect serum levels of F-actin autoantibodies in 93 inpatients with SLE from March 2017 to January 2018 (case group,n=93),72 patients with rheumatoid arthritis (RA) (disease control group) and 83 healthy subjects (healthy control group) were included during the same period.The positive rates of F-actin autoantibodies between the case group and the two control group were compared.Clinical data including SLE disease activity index (SLEDAI),immuno-globulin (lg)G,erythrocyte sedimentation rate (ESR),anti-dsDNA,and antinuclear antibody (ANA) of 93 patients with SLE were collected and the correlation analysis between F-actin autoantibodies units was applied respectively.The diagnostic performance of F-actin autoantibodies in SLE was analyzed by using the receiver operating characteristic curve (ROC).T test,Chi-square test and Spearman/Pearson correlation analysis were applied for statistical analysis.Results The serum levels of F-actin autoantibodies in the SLE case group,disease control group,and healthy control group were (18±13),(12±6),and (11±5) U,respectively,the differences between SLE case group and disease control group,and healthy control group were significant (t=3.163,P=0.001 9;t=4.436,P＜0.01).The positive rates of F-actin autoantibodies were 33％(31/93) in patients with SLE,10％(7/72) in disease control group,and 4％(3/83) in healthy control group.The F-actin autoanti-bodies units in SLE were correlated with SLEDAI,IgG,ESR,anti-dsDNA,and ANA (r=0.273 7,P=0.008 3;r=0.558 7,P＜0.01;r=0.419 9,P=0.000 1,r=0.351 4,P=0.001 1,r=0.460 9,P＜0.01),in which F-actin autoantibodies units showed significant correlation with IgG and ANA.In the ROC curve,the area under the curve(AUC) was 0.62 [95％CI(0.54,0.70)],P=0.001 3.which was statistically significant.When the cut-off value of the F-actin autoantibodies was 14.04 U,the Youden's index (YI) was the largest (YI=0.30),and the sen-sitivity for the diagnosis of SLE was 0.77,the specificity was 0.53.Conclusion The positive rate of F-actin autoantibodies in the serum of patients with SLE is higher than that of RA and healthy controls,so it has certain diagnostic value for SLE.The F-actin autoantibodies units is correlated with both SLEDAI,ESR,and anti-dsDNA,suggesting that F-actin autoantibodies units may be a new biomarker for disease activity assessment of SLE patients.

Coactosin-like protein 1 inhibits neuronal migration during mouse corticogenesis

Coactosin-like protein 1 (Cotl1), a member of the actin-depolymerizing factor (ADF)/cofilin family, was first purified from a soluble fraction of Dictyostelium discoideum cells. Neuronal migration requires cytoskeletal remodeling and actin regulation. Although Cotl1 strongly binds to F-actin, the role of Cotl1 in neuronal migration remains undescribed. In this study, we revealed that Cotl1 overexpression impaired migration of both early- and late-born neurons during mouse corticogenesis. Moreover, Cotl1 overexpression delayed, rather than blocked, neuronal migration in late-born neurons. Cotl1 expression disturbed the morphology of migrating neurons, lengthening the leading processes. This study is the first to investigate the function of Cotl1, and the results indicate that Cotl1 is involved in the regulation of neuronal migration and morphogenesis.

Tannic acid-mediated immune activation attenuates Brucella abortus infection in mice

Brucellosis is an emerging infectious disease affecting humans and animals. In this study, we investigated the in vitro and in vivo effects of tannic acid (TA) against Brucella abortus infection. After infection, F-actin polymerization and mitogen-activated protein kinases (MAPKs) (ERK 1/2 and p38α) phosphorylation were reduced in TA-treated cells compared with that in control cells. The mice were infected via an intraperitoneal route and were orally given TA or phosphate-buffered saline for 14 days. Spleen weights of the TA-treated and control mice were not different; however, splenic proliferation of B. abortus was significantly reduced in the TA-treated group. Immune response analysis showed that, compared with the control group, non-infected TA-treated mice displayed increased levels of interferon-γ (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), and interleukin-10 at 3 days post-infection and a further increase in IFN-γ and MCP-1 at 14 days post-infection. In contrast, compared with the control group, infected TA-treated mice displayed elevated levels of IFN-γ at 3 days post-infection, which continued to increase at 14 days post-infection, as was also observed for tumor necrosis factor. Taken together, the results showing TA activation of cytokine production and inhibition of bacterial proliferation in the host highlight a potential use of TA treatment in the control of Brucella infection.

Mechanisms underlying the urotensin Ⅱ-induced proliferation of and α-smooth muscle actin expression in human dermal fibroblasts / 中华皮肤科杂志

Objective To evaluate the effects of urotensin Ⅱ on cell proliferation of and α-smooth muscle actin (α-SMA) expression in normal human dermal fibroblasts (NFs),and to explore their regulatory mechanisms.Methods NFs were isolated from foreskin tissues and subjected to primary culture in vitro.Reverse transcription PCR and Western blot analysis were performed to measure the mRNA and protein expression of urotensin Ⅱ and its receptor,respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferation of NFs,which were treated with urotensin Ⅱ at different concentrations of 0,10-10,10-9,10-8,10-7 and 10-6 mol/L for 0,6,12,24 and 48 hours separately,and then the optimal concentration and duration of urotensin Ⅱ exposure were selected to be 10-8 mol/L and 24 hours respectively.Some cultured NFs were divided into 5 groups:control group receiving no treatment,U Ⅱ group treated with 10-8 mol/L urotensin Ⅱ,U Ⅱ + nicardipine group treated with 10-8 mol/L urotensin Ⅱ and the calcium channel blocker nicardipine at the concentration of 10-5 mol/L,U Ⅱ + PD98059 group treated with 10-8 mol/L urotensin Ⅱ and the mitogen activated protein kinase (MAPK)inhibitor PD98059 at the concen-tration of 10-5 mol/L,and U Ⅱ + cyclosporine group treated with 10-8 mol/L urotensin Ⅱ and the calcium-dependent protein kinase (CaM PK) inhibitor cyclosporine at the concentration of 10-5 mol/L.After 24-hour treatment,CCK-8 assay was conducted to evaluate the proliferation of NFs in the above groups,real-time fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of α-SMA respectively.Results Urotensin Ⅱ receptor was expressed in NFs,but urotensin Ⅱ was not.The proliferative activity of NFs significantly differed among the control group,U Ⅱ group,U Ⅱ + nicardipine group,U Ⅱ + PD98059 group and U Ⅱ + cyclosporine group (the mean absorbance value at 405 nm:1.036 ± 0.046,1.405 ± 0.158,1.121 ± 0.109,1.192 ± 0.089 and 1.141 ± 0.056,respectively;F =9.587,P ＜ 0.01),and the U Ⅱ group showed significantly higher proliferative activity of NFs compared with the control group,U Ⅱ + nicardipine group,U Ⅱ + PD98059 group and U Ⅱ + cyclosporine group (q =8.263,6.355,4.774 and 5.912,respectively,all P ＜ 0.05).There were significant differences in the mRNA and protein expression of α-SMA among the 5 groups (F =6.351,7.045,both P ＜ 0.01),and the mRNA and protein expression of α-SMA was significantly higher in the U Ⅱ group than in the other 4 groups (all P ＜ 0.05).Conclusion Urotensin Ⅱ may induce the proliferation of and α-SMA expression in NFs through calcium channels,MAPK and CaM PK pathways.

Effects of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes / 中华皮肤科杂志

Objective To evaluate the effect of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes after the plasmid containing the Wnt5A gene is transfected into primary melanocytes.Methods In vitro cultured primary human melanocytes were divided into three groups:blank control group receiving no treatment,negative control group transfected with endotoxin-free pcDNA3.1 (+)empty vector by Lipo3000 in Opti-MEM medium,Wnt5A plasnid group transfected with endotoxin-free pcDNA3.1 (+) vector containing the Wnt5A gene by Lipo3000 in Opti-MEM medium.After the transfection,quantitative PCR (qPCR) was performed to measure the mRNA expression of Wnt5A,ras-related C3 botulinum toxin substrate 1 (Rac1),filamentous actin (F-actin) and β-tubulin,Western blot analysis to determine the protein expression of Wnt5A,receptor tyrosine kinase like orphan receptor 2 (ROR2),Rac1,F-actin and β-tubulin,and an immunofluorescence assay (IFA) to observe the expression of cytoskeletal proteins.Results qPCR showed significant differences in the mRNA expression of the Wnt5A gene and its downstream genes Rac1 and F-actin among the Wnt5A plasmid group,negative control group and blank control group (F =1374.179,112.576,66.458,respectively,all P ＜ 0.01),but there was no significant difference in the mRNA expression of β-tubulin among the three groups (P ＞ 0.05).Additionally,the Wnt5A plasmid group showed significantly higher mRNA expression of Wnt5A,Rac1 and F-actin compared with the blank control group and negative control group (all P ＜ 0.05).As Western blot analysis revealed,compared with the blank control group and negative control group,the Wnt5A plasmid group showed significantly higher Wnt5A protein expression (both P ＜ 0.05),but significantly lower protein expression of Rac 1,ROR2 and F-actin (all P ＜ 0.05).However,no significant difference in β-tubulin protein expression was observed among the three groups (P ＞ 0.05).IFA showed no obvious difference in the fluorescence intensity of β-tubulin or F-actin between the Wnt5A group and the two control groups,but melanocytes showed larger size and increased number of dendrites,and the cytoskeleton changed dramatically with varying fluorescence intensity of F-actin,fuzzy texture,fractured or locally clustered tonofilaments in the Wnt5A group.Conclusion The overexpression of the Wnt5A gene in melanocytes can regulate the mRNA and protein expression of cytoskeletal proteins,nake melanocytes larger and more dendritic,and cause changes in the cytoskeleton,which may facilitate the transportation of melanosomes,and participate in the occurrence of hyperpigmented diseases.

Effects of different degrees of intermittent hypoxia on the activation and the extracellular matrix secretion of mouse lung fibroblast cells / 天津医药

Objective To explore the effects of different degrees of intermittent hypoxia (IH) on the activation and the secretion of extracellular matrix in MLg lung fibroblast cell line. Methods MLg lung fibroblast cells in logarithmic growth phase were exposed for 5%O2 for 100 seconds and 21%O2 for 120 seconds in 1 h, 4 h and 8 h groups (IH1, IH4 and IH8) and normoxia group (21%O2 for 8 h, N group). The cells in each group were collected at the end of experiment. Real-time PCR was used to measure the mRNA expression levels ofα-SMA and typeⅠcollagen (COL1) A1, and Western blot assay was used to detect the protein expression levels ofα-SMA and COL1. Results The mRNA and protein expression levels ofα-SMA and COL1 were significantly increased in IH1, IH4 and IH8 groups than those in N group (all P < 0.05). Furthermore, expression levels of α-SMA and COL1 showed a time-dependent increase with IH exposure time. Conclusion The intermittent hypoxia can promote the cell activation and the extracellular matrix secretion of mouse lung fibroblast cells, which may be related with the oxidative stress.

Substrate roughness induces the development of defective E-cadherin junctions in human gingival keratinocytes

PURPOSE: The entry of bacteria or harmful substances through the epithelial seal of human gingival keratinocytes (HGKs) in the junctional epithelium (JE) is blocked by specialized intercellular junctions such as E-cadherin junctions (ECJs). However, the influence of roughened substrates, which may occur due to apical migration of the JE, root planing, or peri-implantitis, on the development of the ECJs of HGKs remains largely unknown. METHODS: HGKs were cultured on substrates with varying levels of roughness, which were prepared by rubbing hydrophobic polystyrene dishes with silicon carbide papers. The activity of c-Jun N-terminal kinase (JNK) was inhibited with SP600125 or by transfection with JNK short hairpin RNA. The development of intercellular junctions was analyzed using scanning electron microscopy or confocal laser scanning microscopy after immunohistochemical staining of the cells for E-cadherin. The expression level of phospho-JNK was assessed by immunoblotting. RESULTS: HGKs developed tight intercellular junctions devoid of wide intercellular gaps on smooth substrates and on rough substrates with low-nanometer dimensions (average roughness [Ra]=121.3±13.4 nm), although the ECJs of HGKs on rough substrates with low-nanometer dimensions developed later than those of HGKs on smooth substrates. In contrast, HGKs developed short intercellular junctions with wide intercellular gaps on rough substrates with mid- or high-nanometer dimensions (Ra=505.3±115.3 nm, 867.0±168.6 nm). Notably, the stability of the ECJs was low on the rough substrates, as demonstrated by the rapid destruction of the cell junction following calcium depletion. Inhibition of JNK activity promoted ECJ development in HGKs. JNK was closely associated with cortical actin in the regulation of ECJs in HGKs. CONCLUSIONS: These results indicate that on rough substrates with nanometer dimensions, the ECJs of HGKs develop slowly or defectively, and that this effect can be reversed by inhibiting JNK.

Mechanism of α-naphthyl isothiocyanate inducing cholestatic hepatitis: a preliminary study / 临床肝胆病杂志

ObjectiveTo investigate the mechanism of α-naphthyl isothiocyanate (ANIT) inducing cholestatic hepatitis. MethodsA total of 60 healthy male Sprague-Dawley rats were randomly divided into normal control group (N group with 30 rats) and model group (ANIT group with 30 rats), and each group was further divided into three subgroups according to different time phases after gavage (at 24, 48, and 72 hours after gavage), with 10 rats in each group. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), and total bilirubin (TBil) were measured, and the level of malondialdehyde (MDA) in the liver was measured. A light microscope was used to observe the pathological changes in the liver, and a transmission electron microscope was used to observe the changes in the ultrastructure of the bile canaliculus. A confocal laser scanning microscope was used to analyze the mean fluorescence intensity of phalloidine labeled by fluorescein isothiocyanate in order to determine the change in the level of fibrous actin (F-actin). ResultsAfter the administration of ANIT by gavage, the model group showed significant increases in the serum levels of ALT, AST, GGT, and TBil and the level of MDA in liver tissue compared with the normal control group. The model group also showed damage in the structure of the hepatic lobules, hyaline degeneration of hepatocytes, spotted, patchy, and focal necrotic lesions, and neutrophil infiltration mostly confined to the hepatocytes around the bile duct, proliferation of biliary epithelial and fibrous tissues, widened perisinusoidal spaces, dilation of the bile canaliculi, extensive shedding of microvilli, and formation of bile thrombi in the bile capillaries, as well as a low fluorescence intensity of F-actin. The above changes were the most obvious at 48 hours; recovery began at 72 hours, but significant differences were still seen between the two groups. ConclusionANIT can cause lipid peroxidation in liver tissue and lead to degeneration and necrosis of liver cells, damage of microvilli of the bile canaliculi, and formation of bile thrombi, as well as reduction in the expression of F-actin in the bile canaliculi. Therefore, it affects the function of the liver to secrete bile and causes cholestatic hepatitis.

Treponema pallidum membrane protein Tpp47 regulates the permeability of vascular endothelial cells via the RhoA/ROCK signal pathway: an experimental study / 中华皮肤科杂志

Objective To investigate mechanisms underlying the regulation of the permeability of vascular endothelial cells by the Treponema pallidum membrane protein Tpp47.Methods Human umbilical vascular endothelial cell (HUVEC) monolayers were established as a model,and were directly cultured with the presence of recombinant Tpp47 protein (rTpp47-treated group),or boiled and inactivated rTpp47 (negative control group).Some HUVEC monolayers,which were pretreated with the RhoA/ROCK signal pathway inhibitor Y-27632 for 30 minutes and then cultured with the presence of rTpp47,served as the pretreatment group.After 1-and 4-hour additional culture,enzymelinked immunosorbent assay (ELISA) was performed to estimate the permeability of these cell monolayers to horseradish peroxidase (HRP).After 12 hours of culture,rhodamine-phalloidin was used to stain cytoskeletal proteins,and confocal laser scanning microscopy was performed to observe the arrangement of the cytoskeletal protein F-actin.Western-blot analysis was conducted to measure the expressions of RhoA in HUVECs treated with rTpp47 or inactivated rTpp47.Results The supernatant level of HRP (expressed as the absorbance value at 450 nm) was significantly higher in the rTpp47-treated group than in the negative control group (0.81 ± 0.10 vs.0.39 ± 0.09,P ＜ 0.05),but no significant difference was observed between the pretreatment group (0.51 ± 0.10) and rTpp47-treated group or negative control group (both P ＞ 0.05) after 1-hour culture.Similarly,the rTpp47-treated group showed significantly increased levels of HRP compared with the pretreatment group and negative control group (2.31-± 0.14 vs.1.21 ± 0.12 and 0.73 ± 0.12,both P ＜ 0.05),while there was no significant difference between the pretreatment group and negative control group after 4-hour culture.The expression of RhoA in HUVECs treated with rTpp47 was significantly higher than that in those treated with inactivated rTpp47.Confocal laser scanning microscopy showed that rTpp47 treatment led to the rearrangement of F-actin in HUVECs followed by the formation of stress fibers in cytoplasm,while Y-27632 could partly inhibit the rearrangement of F-actin.Conclusion The recombinant Treponema pallidum membrane protein Tpp47 can regulate the permeability of vascular endothelial cells through the RhoA/ROCK signal pathway.

α-Smooth Muscle Actin and ACTA2 Gene Expressions in Vasculopathies

ABSTRACT α-smooth muscle actin, encoded by ACTA2 gene, is an isoform of the vascular smooth muscle actins, typically expressed in the vascular smooth muscle cells contributing to vascular motility and contraction. ACTA2 gene mutations cause a diversity of diffuse vasculopathies such as thoracic aortic aneurysms and dissections as well as occlusive vascular diseases, including premature coronary artery disease and ischemic stroke. Dynamics of differentiation-specific α-smooth muscle actin in arterial smooth muscle cells and proliferation of the proteins have been well described. Although a variety of research works have been undertaken in terms of modifications of α-smooth muscle actin and mutations of ACTA2 gene and myosin, the underlying mechanisms towards the pathological processes by way of gene mutations are yet to be clarified. The purpose of the present article is to describe the phenotypes of α-smooth muscle actin and implications of ACTA2 mutations in vasculopathies in order to enhance the understanding of potential mechanisms of aortic and coronary disorders.