Experimental acrylonitrile butadiene styrene and polyamide evisceration implant: a rabbit clinical and histopathology study / Estudo clínico e histopatológico em coelhos eviscerados com utilização de implantes de poliamida e acrilonitrila butadieno estireno

The purpose of this study was to evaluate acrylonitrile butadiene styrene (ABS) and polyamide implants in rabbits submitted to evisceration at the macroscopic and microstructure level and to assess clinical response and histopathological changes as well. For the experimental study implants of 12mm diameter were prepared by rapid prototyping, weighed and the outer and inner surfaces evaluated macroscopically and by electron microscopy. In addition, a compression test was performed and ultrastructural damage was then determined. After evisceration of the left eyeball, nine New Zealand rabbits received ABS implants and nine others received polyamide implants. The animals were assessed daily for 15 days after surgery and every seven days until the end of the study (90 days). Histopathological evaluation was performed at 15, 45 and 90 days after surgery. The ABS implants weighed approximately 0.44g, while the polyamide ones weighed 0.61g. Scanning electron microscopy demonstrated that the ABS implants had regular-sized, equidistant micropores, while the polyamide ones showed micropores of various sizes. The force required to fracture the ABS implant was 14.39 ±0.60 Mpa, while for the polyamide one, it was 16.80 ±1.05 Mpa. Fifteen days after surgery, we observed centripetal tissue infiltration and scarce inflammatory infiltrate. Implants may be used in the filling of anophthalmic cavities, because they are inert, biocompatible and allow tissue integration.(AU)

Avaliou-se a macroscopia, a microestrutura, a resposta clínica e histopatológica de implantes de acrilonitrila butadieno estireno (ABS) e poliamida em coelhos submetidos à evisceração. Para o estudo experimental, os implantes foram elaborados por meio de prototipagem rápida, com 12mm de diâmetro, pesados e tiveram suas superfícies avaliadas macroscopicamente e por microscopia eletrônica de varredura. Adicionalmente, foi realizado ensaio de compressão para determinar a força necessária para fraturar os implantes. Após a evisceração do olho esquerdo, nove coelhos da raça Nova Zelândia receberam implantes de poliamida e nove outros receberam implantes de ABS. Os animais foram avaliados diariamente nos primeiros 15 dias após a cirurgia e a cada sete dias até o fim do período experimental (90 dias). Avaliação histopatológica foi realizada aos 15, 45 e 90 dias após a cirurgia. Os implantes de ABS pesaram 0,44g, e os de poliamida 0,61g. A microscopia eletrônica de varredura demonstrou que os implantes de ABS eram formados por microporos equidistantes, enquanto os de poliamida apresentavam microporos de vários tamanhos. A força necessária para fraturar os implantes de ABS foi de 14.39±0.60 Mpa, enquanto para os de poliamida foi de 16.80±1.05Mpa. Quinze dias após a cirurgia, foi observada infiltração fibrovascular centrípeta. Os implantes podem ser utilizados para correção de cavidades anoftálmicas por se mostrarem inertes, biocompatíveis e permitirem a infiltração tecidual.(AU)

Advances in metabolic engineering of Escherichia coli for isoprene biosynthesis / 生物工程学报

As an important industrial chemical, isoprene is mainly used as a precursor for synthetic rubbers. In addition, it also has wide applications in the field of pharmaceutical and chemical intermediates, food, adhesives and aviation fuel. Compared with conventional petrochemical routes, production of isoprene in microbial systems has been the research focus considering environment friendly and sustainable development features. This article summarizes the metabolic pathways and key enzymes of isoprene biosynthesis, reviews current methods and strategies in improving isoprene production of Escherichia coli, and also gives some basic ideas and expectation.

Extracellular signal-regulated kinase signaling pathway regulates the endothelial differentiation of periodontal ligament stem cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of extracellular signal-regulated kinase (ERK) signaling pathway on the endothelial differentiation of periodontal ligament stem cells (PDLSC).</p><p><b>METHODS</b>Human PDLSC was cultured in the medium with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF) to induce endothelial differentiation. Endothelial inducing cells was incubated with U0126, a specific p-ERK1/2 inhibitor. PDLSC from one person were randomly divided into four groups: control group, endothelial induced group, endothelial induced+DMSO group and endothelial induced+U0126 group. The protein expression of the p-EKR1/2 was analyzed by Western blotting at 0, 1, 3, 6 and 12 hours during endonthelial induction. The mRNA expressions of CD31, VE-cadherin, and VEGF were detected by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) after a 7-day induction. The proportion of CD31(+) to VE-cadherin(+) cells was identified by flow cytometry, and the ability of capillary-like tubes formation was detected by Matrigel assay after a 14-day induction. The measurement data were statistically analyzed.</p><p><b>RESULTS</b>Phosphorylated ERK1/2 protein level in PDLSC was increased to 1.24±0.12 and 1.03±0.24 at 1 h and 3 h respectively, during the endothelial induction (P<0.01). The mRNA expressions of CD31 and VEGF in induced+U0126 group were decreased to 0.09±0.18 and 0.49±0.17, which were both significantly different with those in induced group (P<0.05). The proportion of CD31(+) to VE-cadherin(+) cells of induced+U0126 group were decreased to 5.22±0.85 and 3.56±0.87, which were both significantly different with those in induced group (P<0.05). In Matrigel assay, the branching points, tube number and tube length were decreased to 7.0±2.7, 33.5±6.4, and (15 951.0±758.1) pixels, which were all significantly different with those in induced group (P<0.05).</p><p><b>CONCLUSIONS</b>The endothelial differentiation of PDLSC is positively regulated by ERK signaling pathway. Inhibition of ERK1/2 phosphorylation could suppress endothelial differentiation of PDLSC.</p>

Prevalência de dor crônica e sua associação com a situação sociodemográfica e atividade física no lazer em idosos de Florianópolis, Santa Catarina: estudo de base populacional / Prevalence of chronic pain and its association with the sociodemographic situation and physical activity in leisure of elderly in Florianópolis, Santa Catarina: population-based study

OBJETIVO: Estimar a prevalência de dor crônica e sua associação com a situação socioeconômica, demográfica e atividade física no lazer em idosos. MÉTODOS: Este estudo é parte do inquérito epidemiológico e transversal de base populacional e domiciliar EpiFloripa Idoso 2009-2010 realizado com 1.705 idosos (≥ 60 anos), residentes em Florianópolis, Santa Catarina. A partir da resposta afirmativa de dor crônica, foram investigadas as associações com as variáveis obtidas por meio de entrevista estruturada. Realizou-se a estatística descritiva, incluindo cálculos de proporções e intervalos de confiança 95% (IC95%). Na análise bruta e ajustada, empregou-se regressão de Poisson, estimando-se as razões de prevalência, com intervalos de confiança de 95% e valores p ≤ 0,05. RESULTADOS: Dentre os idosos investigados, 29,3% (IC95% 26,5 - 32,2) relataram dor crônica. Na análise ajustada, observou-se que as variáveis sexo feminino, menor escolaridade e pior situação econômica ficaram associadas significativamente com maior prevalência de dor crônica; ser fisicamente ativo no lazer ficou associado significativamente com menor prevalência do desfecho. CONCLUSÕES: Percebe-se que a dor crônica é um agravo que acomete considerável parcela de idosos, havendo desigualdades sociais na sua frequência e sendo beneficamente afetada pela atividade física no lazer. É necessário que políticas públicas de saúde subsidiem programas multidisciplinares de controle da dor incluindo a prática regular de atividade física, voltada especificamente à promoção da saúde do idoso, evitando assim que a dor crônica comprometa a qualidade de vida desta população. .

OBJECTIVE: To estimate the prevalence of chronic pain and its association with socioeconomic and demographic status, and leisure physical activity in the elderly population. METHODS: This study is part of an epidemiological cross-sectional population-based household survey called EpiFloripa Elderly 2009-2010, which was conducted with 1,705 elderly individuals (≥ 60 years) residents of Florianópolis, Santa Catarina. From the positive response to chronic pain, the associations with the variables were investigated through a structured interview. Descriptive statistics were conducted, including ratio calculation and 95% confidence intervals. In crude and adjusted analysis, Poisson regression was utilized, estimating prevalence ratios, with 95% confidence intervals and ≤ 0.05 p-values. RESULTS: Among the subjects, 29.3% (IC95% 26.5 - 32.2) reported chronic pain. Adjusted analysis showed that being female, having less years of schooling, and being in worse economic situation were significantly associated with a higher prevalence of chronic pain. Being physically active during leisure time was significantly associated with lower prevalence of the outcome. CONCLUSIONS: Therefore, it is clear that chronic pain affects a considerable amount of elderly individuals. Social inequalities are a harmful influence in these individuals' quality of life, inasmuch as those inequalities increase the frequency with which chronic pain afflicts them. At the same time, physical activity during leisure time decreases chronic pain frequency. It is fundamental that public health policies subsidize multidisciplinary pain management programs, which should include health targeted physical activity for the elderly, thus preventing the decrease in quality of life that chronic pain brings to this population. .

Improving isoprene production by engineered heterologous mevalonate pathway in Escherichia coli / 生物工程学报

Isoprene is an important precursor of synthetic rubber material. In our previous study, metabolic engineered Escherichia coli strain (BW-01) was constructed and used to produce isoprene. Based on the theory of protein budget, using synthetic biology strategies including the increased copy number of genes and rare codons, we regulated the expression of key enzyme to improve isoprene production in Escherichia coli strain. Under shake-flask conditions, isoprene productivity of the engineered strain (BW-07) increased by 73% compared with BW-01, reached 761.1 mg/L. It provides a reference for further studies.

Downregulation of Aquaporin 4 Expression through Extracellular Signal-regulated Kinases1/2 Activation in Cultured Astrocytes Following Scratch-injury / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the role of extracellular signal-regulated kinase1/2 (ERK1/2) pathway in the regulation of aquaporin 4 (AQP4) expression in cultured astrocytes after scratch-injury.</p><p><b>METHODS</b>The scratch-injury model was produced in cultured astrocytes of rat by a 10-μL plastic pipette tip. The morphological changes of astrocytes and lactate dehydrogenase (LDH) leakages were observed to assess the degree of scratch-injury. AQP4 expression was detected by immunofluorescence staining and Western blot, and phosphorylated-ERK1/2 (p-ERK1/2) expression was determined by Western blot. To explore the effect of ERK1/2 pathway on AQP4 expression in scratch-injured astrocytes, 10 µmol/L U0126 (ERK1/2 inhibitor) was incubated in the medium at 30 min before the scratch-injury in some groups.</p><p><b>RESULTS</b>Increases in LDH leakage were observed at 1, 12, and 24 h after scratch-injury, and AQP4 expression was reduced simultaneously. Decrease in AQP4 expression was associated with a significant increase in ERK1/2 activation. Furthermore, pretreatment with U0126 blocked both ERK1/2 activation and decrease in AQP4 expression induced by scratch-injury.</p><p><b>CONCLUSION</b>These results indicate that ERK1/2 pathway down-regulates AQP4 expression in scratch-injured astrocytes, and ERK1/2 pathway might be a novel therapeutic target in reversing the effects of astrocytes that contribute to traumatic brain edema.</p>

Epidemiology of cervical cancer in Colombia / Epidemiología del cáncer cervical en Colombia

Objective. To describe the incidence, mortality, time trends and prognostic factors for cervical cancer in Cali, Colombia, and to review the molecular epidemiological evidence showing that HPV is the major and necessary cause of cervical cancer and the implications of this discovery for primary and secondary prevention. Materials and methods. Incidence rates of cervical cancer during a 45-year period (1962-2007) were estimated based on the population-based cancer registry of Cali and the mortality statistics from the Municipal Health Secretariat of Cali. Prognostic factors were estimated based on relative survival. Review of the molecular epidemiological evidence linking HPV to cervical cancer was focused on the studies carried out in Cali and in other countries. Results. Incidence rates of squamous cell carcinoma (SCC) declined from 120.4 per 100 000 in 1962-1966 to 25.7 in 2003-2007 while those of adenocarcinoma increased from 4.2 to 5.8. Mortality rates for cervical cancer declined from 18.5 in 1984-1988 to 7.0 per 100 000 in 2009-2011. Survival was lower in women over 65 years of age and in clinical stages 3-4. Review of the molecular epidemiological evidence showed that certain types of HPV are the central and necessary cause of cervical cancer. Conclusions. A decline in the incidence and mortality of SCC and an increase in the incidence of adenocarcinoma during a 45-year period was documented in Cali, Colombia.

Objetivos. Describir la tendencia temporal de la incidencia, mortalidad y los factores pronósticos del cáncer de cuello uterino en Cali, Colombia, y revisar la evidencia epidemiológica molecular que muestra que el VPH es la causa principal y necesaria del cáncer cervical y las implicaciones de este descubrimiento para la prevención primaria y secundaria. Material y métodos. Se estimaron las tasas de incidencia de cáncer de cuello uterino durante un periodo de 45 años (1962-2007) con la información del registro poblacional de cáncer de Cali y las de mortalidad con las estadísticas de la Secretaría de Salud Municipal de Cali. Los factores pronósticos se estimaron sobre la base de la supervivencia relativa. La revisión de la evidencia epidemiológica molecular que une el VPH con el cáncer cervical se centró en los estudios llevados a cabo en Cali y en otros países. Resultados. Las tasas de incidencia de carcinoma de células escamosas (SCC) por 100000 se redujeron desde 120.4 en 1962-66 a 25.7 en 2003-07, mientras que las de adenocarcinoma aumentaron desde 4.2 hasta 5.8. Las tasas de mortalidad por cáncer de cuello uterino por 100000 se redujeron desde 18.5 en 1984-88 a 7.0 en 2009-11. La supervivencia fue menor en las mujeres de más de 65 años de edad y en los estadios clínicos 3-4. La revisión de la evidencia epidemiológica molecular mostró que ciertos tipos de VPH son la causa central y necesaria del cáncer cervical. Conclusiones. Se documentó la disminución de la incidencia y la mortalidad por SCC y un aumento en la incidencia de adenocarcinoma durante un periodo de 45 años en Cali, Colombia.

Incidencia, mortalidad y supervivencia al cáncer de próstata en Cali, Colombia, 1962-2011 / Prostate cancer in Cali, Colombia, 1962-2011: incidence, mortality and survival

Objetivo. Analizar la tendencia de las tasas de incidencia, mortalidad y supervivencia al cáncer de próstata (CP) en el periodo 1962-2011. Material y métodos. Con base en el Registro Poblacional de Cáncer en Cali (Colombia) y los registros de mortalidad de la Secretaría de Salud Pública Municipal se analizaron las tendencias de las tasas de incidencia, mortalidad y supervivencia por CP en Cali durante el periodo 1962-2011. Resultados. La incidencia de cáncer de próstata aumentó vertiginosamente entre 1986 y 2002 (Annual Percent Change APC 6.2%) y se estabilizó posteriormente. La mortalidad disminuyó desde 1997 en los mayores de 70 años, mientras que en el grupo de 50-69 años fue a partir de 1981. La supervivencia relativa a cinco años fue 69.8% (IC95% 67.5-72.0) y se asoció de manera significativa con la edad, periodo de diagnóstico y estrato socioeconómico. Conclusión. El incremento en la incidencia del CP coincide temporalmente con la implementación del antígeno específico de próstata (PSA por sus siglas en inglés) en Cali; hay evidencia de mejoría en la supervivencia en el CP y disminución en su mortalidad.

Objective. To analyze the trend in prostate cancer survival, incidence and mortality rates in Cali, Colombia from 1962 to 2011. Materials and methods. Based on the Cancer Registry of Cali, Colombia and the mortality registry of the City's Public Health Secretary, incidence, mortality age-standardized rates and relative survival were calculated during 1962-2011. Results. Prostate cancer incidence rates increased sharply between 1986 and 2002 (APC: 6.21%) and then leveled off. Mortality diminished in 1997 in men older than 70 years-old while in men aged 50-69 years declined since 1981. The 5-year-relative-survival was 69.8% (CI95% 67.5-72.0) and it was significantly associated with age, quinquennial period of diagnosis and socioeconomic strata. Conclusion. The increase in incidence rates of prostate cancer in time coincides with the implementation of the PSA in Cali. There is evidence of improvement in prostate cancer survival, and decreased prostate cancer mortality.

Incidencia, mortalidad y supervivencia por cáncer colorrectal en Cali, Colombia, 1962-2012 / Colorectal cancer incidence, mortality and survival in Cali, Colombia, 1962-2012

Objetivo. Estudiar el comportamiento del cáncer colorrectal (CCR) en Cali, Colombia, durante el periodo 1962-2012, utilizando la información del Registro Poblacional de Cáncer de Cali y de la Secretaría de Salud Municipal de Cali. Material y métodos. Análisis ecológico de series de tiempo para estudiar la tendencia de las tasas de incidencia (1962-2007) y mortalidad por CCR (1984-2012); y un análisis de supervivencia de la serie de casos de CCR registrados en Cali entre 1995-2004. La tendencia temporal de las tasas de incidencia (TIEE) y mortalidad (TMEE) estandarizadas por edad se estudió mediante el cambio medio anual (APC por sus siglas en inglés). Se estimó la supervivencia relativa a cinco años y se hizo un análisis múltiple mediante el modelo de regresión de riesgos proporcionales de Cox. Resultados. Durante el periodo 1962-2007, las TIEE de CCR aumentaron en los hombres y mujeres residentes en Cali; APC= 2.6 (IC95% 2.2-3.0) y APC= 2.2% (IC95% 1.8-2.7), respectivamente. Entre 1984-2012 las TMEE permanecieron estables en las mujeres y en los hombres aumentaron en todos los grupos de edad; APC=1.8 (IC95% 0.8-2.8). La supervivencia relativa a cinco años fue independiente del sexo y aumentó de 29.7% en 1995-1999 a 39.8% durante 2000-2004. El riesgo de morir por CCR fue mayor en las personas de estratos socioeconómicos (ESE) bajos frente a los ESE altos, HR= 2.1 (IC95% 1.7-2.6); en los mayores de 70 años frente a los <50, HR= 2.4 (IC95% 1.9-2.9) y en el periodo 1995-1999 frente al 2000-2004 HR=1.5 (IC95% 1.3-1.7). Conclusión. El cáncer de colon y recto está ocupando un lugar preponderante entre los cánceres de mayor importancia en Cali, Colombia.

Objective. To study the colorectal cancer (CRC) behavior in Cali, Colombia, during the 1963-2012 period using data from the Population-based Cancer Registry of Cali and the Municipal Health Secretariat of Cali. Materials and methods. An ecological time series analysis to study the CRC incidence (1962-2007) and mortality (1984-2012) rate trends; and a survival analysis of CRC cases registered in Cali between 1995 and 2004 were conducted. The age-standardized temporal trend of incidence (I-ASR) and mortality (M-ASR) rates were studied using an annual percent change (APC). The 5-year relative survival was estimated and a multivariate analysis was performed using the Cox proportional hazard regression model. Results. During the 1962-2007 period, CRC TTIR increased in men and women living in Cali [APC= 2.6 (95% CI 2.2-3.0) and APC= 2.2% (95% CI 1.8-2.7), respectively]. In the 1984-2012 period, the TTMR remained stable in women but increased in men in all age groups [APC= 1.8 (95% CI 0.8-2.8)]. The 5-year relative survival was independent of sex and increased from 29.7% in 1995-1999 to 39.8% in 2000-2004. The risk of dying from CRC was higher in people of lower socio-economic status (SES) vs higher SES [HR= 2.1 (95% CI: 1.7-2.6)], among people older than 70 years of age vs younger than 50 years [HR= 2.4 (95% CI: 1.9-2.9)], and for the 1995-1999 period vs 2000-2004 period [HR= 1.5 (95% CI 1.3-1.7)]. Conclusion. CRC is beginning to take a prominent place among the most important cancers in Cali, Colombia.

Cost analysis of different cervical cancer screening strategies in Mexico / Análisis de costos de distintas estrategias de tamizaje para cáncer cervical en México

Objective. To compare the costs and number of undetected cases of four cervical cancer screening strategies (CCSS) in Mexico. Materials and methods. We estimated the costs and outcomes of the following CCSS: a) conventional Papanicolaou smear (Pap) alone; b) high-risk human papilloma virus testing (HR-HPV) as primary screening with Pap as reflex triage; c) HR-HPV as primary screening with HPV-16/18 typing, liquid-based cytology (LBC) and immunostaining for p16/Ki67 testing as reflex triage, and d) co-testing with HR-HPV and LBC with HPV-16/18 typing and immunostaining for p16/Ki67 as reflex triage. The outcome of interest was high-grade cervical lesions or cervical cancer. Results. HR-HPV testing, HPV typing, LBC testing and immunostaining is the best alternative because it is the least expensive option with an acceptable number of missed cases. Conclusions. The opportunity costs of a poor quality CCSS is many false negatives. Combining multiple tests may be a more cost-effective way to screen for cervical cancer in Mexico.

Objetivo. Comparar los costos y los casos no detectados de cuatro estrategias de tamizaje de cáncer cervical (ETCC) en México. Material y métodos. Se estimaron los costos y resultados en salud de las siguientes ETCC: a) citología convencional como único procedimiento de tamizaje; b) detección de virus del papiloma humano de alto riesgo (VPH-AR) como tamizaje primario y citología convencional como procedimiento de triage; c) detección de VPH-AR como tamizaje primario y tipificación de VPH-16/18, citología en base líquida e inmunotinción para p16/Ki67 como procedimientos de triage, y d) evaluación conjunta con VPH-AR y citología en base líquida como tamizaje primario y tipificación de VPH-16/18 e inmunotinción para p16/Ki67 como procedimientos de triage. El resultado en salud analizado fueron los casos de neoplasia intraepitelial cervical (CIN 2/3) o cáncer cervical detectados. Resultados. La ETCC basada en la detección de VPH-AR como prueba primaria y seguida de la tipificación de VPH-16/18, la citología en base líquida y la inmunotinción para p16/Ki67 como procedimientos de triage es la mejor alternativa, ya que es la menos costosa y la que tuvo un nivel aceptable de casos perdidos. Conclusiones. El costo de oportunidad de una ETCC de mala calidad es un alto número de falsos negativos. La combinación seriada de varias pruebas de tamizaje y triage puede ser una alternativa costo-efectiva para la detección oportuna de cáncer cervical en México.

Dieta y cáncer gástrico en México y en el mundo / Diet and gastric cancer in Mexico and in the world

El cáncer gástrico (CG) es la cuarta causa de muerte por cáncer a nivel global. La dieta y el consumo de alcohol y tabaco, además de la infección por Helicobacter pylori determinan un gran número de casos de esta neoplasia. Algunos alimentos contienen sustancias que podrían influir en el proceso de carcinogénesis gástrica, aunque los mecanismos subyacentes no están completamente dilucidados. En México y el mundo, la disminución en el consumo de frutas, vegetales no feculentos y allium, leguminosas y alimentos fuente de selenio, así como el aumento en el consumo de sal, alimentos salados, salmuera y ahumados, chile, carnes procesadas y asadas o a la parrilla se han asociado respectivamente con un aumento de riesgo de CG. Con la evidencia disponible, se podrían desarrollar y evaluar programas para la prevención y control del CG.

Gastric cancer (GC) is the fourt leading cause of cancer death at global level. Diet, alcohol and tobacco, in addition to Helicobacter pylori infection, account for a large number of cases. Some substances contained in foods may influence GC carcinogenesis process; however, the underlying mechanisms have not been fully elucidated. In Mexico and worldwide, a low intake of fruits, non-starchy and allium vegetables, pulses, and foods containing selenium, as well as high intake of salt, salty, salted and smoked foods, chili pepper, processed and grilled/barbecued meats, have been respectively associated with an increased risk of GC. Based on the available evidence, programs for GC prevention and control could be developed and evaluated.

Vitamin C induces periodontal ligament progenitor cell differentiation via activation of ERK pathway mediated by PELP1

The differentiation of periodontal ligament (PDL) progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone. Vitamin C (VC), a water-soluble nutrient that cannot be biosynthesized by humans, is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling. Therefore, the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level. We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents. During the process, VC preferentially activated ERK1/2 but did not affect JNK or p38. Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2. ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation. PELP1, a nuclear receptor co-regulator, was up-regulated under VC treatment. PELP1 knockdown inhibited ERK phosphorylation. The overexpression of PELP1 had a positive relationship with Runx2 expression. Taken together, we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis. Our finding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.

Insulin Like Growth Factor Binding Protein-5 Regulates Excessive Vascular Smooth Muscle Cell Proliferation in Spontaneously Hypertensive Rats via ERK 1/2 Phosphorylation

Insulin-like growth factor binding proteins (IGFBPs) are important components of insulin growth factor (IGF) signaling pathways. One of the binding proteins, IGFBP-5, enhances the actions of IGF-1, which include the enhanced proliferation of smooth muscle cells. In the present study, we examined the expression and the biological effects of IGFBP-5 in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). The levels of IGFBP-5 mRNA and protein were found to be higher in the VSMC from SHR than in those from WKY. Treatment with recombinant IGFBP-5-stimulated VSMC proliferation in WKY to the levels observed in SHR. In the VSMCs of WKY, incubation with angiotensin (Ang) II or IGF-1 dose dependently increased IGFBP-5 protein levels. Transfection with IGFBP-5 siRNA reduced VSMC proliferation in SHR to the levels exhibited in WKY. In addition, recombinant IGFBP-5 significantly up-regulated ERK1/2 phosphorylation in the VSMCs of WKY as much as those of SHR. Concurrent treatment with the MEK1/2 inhibitors, PD98059 or U0126 completely inhibited recombinant IGFBP-5-induced VSMC proliferation in WKY, while concurrent treatment with the phosphatidylinositol-3 kinase inhibitor, LY294002, had no effect. Furthermore, knockdown with IGFBP-5 siRNA inhibited ERK1/2 phosphorylation in VSMC of SHR. These results suggest that IGFBP-5 plays a role in the regulation of VSMC proliferation via ERK1/2 MAPK signaling in hypertensive rats.

High Glucose Induces Connective Tissue Growth Factor Expression and Extracellular Matrix Accumulation in Rat Aorta Vascular Smooth Muscle Cells Via Extracellular Signal-Regulated Kinase 1/2

Connective tissue growth factor (CTGF) is a potent pro-fibrotic factor, which is implicated in fibrosis through extracellular matrix (ECM) induction in diabetic cardiovascular complications. It is an important downstream mediator in the fibrotic action of transforming growth factor beta (TGFbeta) and is potentially induced by hyperglycemia in human vascular smooth muscle cells (VSMCs). Therefore, the goal of this study is to identify the signaling pathways of CTGF effects on ECM accumulation and cell proliferation in VSMCs under hyperglycemia. We found that high glucose stimulated the levels of CTGF mRNA and protein and followed by VSMC proliferation and ECM components accumulation such as collagen type 1, collagen type 3 and fibronectin. By depleting endogenous CTGF we showed that CTGF is indispensable for the cell proliferation and ECM components accumulation in high glucose-stimulated VSMCs. In addition, pretreatment with the MEK1/2 specific inhibitors, PD98059 or U0126 potently inhibited the CTGF production and ECM components accumulation in high glucose-stimulated VSMCs. Furthermore, knockdown with ERK1/2 MAPK siRNA resulted in significantly down regulated of CTGF production, ECM components accumulation and cell proliferation in high glucose-stimulated VSMCs. Finally, ERK1/2 signaling regulated Egr-1 protein expression and treatment with recombinant CTGF reversed the Egr-1 expression in high glucose-induced VSMCs. It is conceivable that ERK1/2 MAPK signaling pathway plays an important role in regulating CTGF expression and suggests that blockade of CTGF through ERK1/2 MAPK signaling may be beneficial for therapeutic target of diabetic cardiovascular complication such as atherosclerosis.

Role of Extracellular Signal-Regulated Kinase 1/2 and Reactive Oxygen Species in Toll-Like Receptor 2-Mediated Dual-Specificity Phosphatase 4 Expression / 영남의대학술지

BACKGROUND: Toll-like receptors (TLRs) are well-known pattern recognition receptors. Among the 13 TLRs, TLR2 is the most known receptor for immune response. It activates mitogen-activated protein kinases (MAPKs), which are counterbalanced by MAPK phosphatases [MKPs or dual-specificity phosphatases (DUSPs)]. However, the regulatory mechanism of DUSPs is still unclear. In this study, the effect of a TLR2 ligand (TLR2L, Pam3CSK4) on DUSP4 expression in Raw264.7 cells was demonstrated. METHODS: A Raw264.7 mouse macrophage cell line was cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 1% antibiotics (100 U/mL penicillin and 100 g/mL streptomycin) at 37degrees C in 5% CO2. TLR2L (Pam3CSK4)-mediated DUSP4 expressions were confirmed with RT-PCR and western blot analysis. In addition, the detection of reactive oxygen species (ROS) was measured with lucigenin assay. RESULTS: Pam3CSK4 induced the expression of DUSP1, 2, 4, 5 and 16. The DUSP4 expression was also increased by TLR4 and 9 agonists (lipopolysaccharide and CpG ODN, respectively). Pam3CSK4 also induced ERK1/2 phosphorylation and ROS production, and the Pam3CSK4-induced DUSP4 expression was decreased by ERK1/2 (U0126) and ROS (DPI) inhibitors. U0126 suppressed the ROS production by Pam3CSK4. CONCLUSION: Pam3CSK4-mediated DUSP4 expression is regulated by ERK1/2 and ROS. This finding suggests the physiological importance of DUSP4 in TLR2-mediated immune response.

Regulation of isoprenoid pathway for enhanced production of linalool in Saccharomyces cerevisiae / 生物工程学报

Linalool is an important monoterpene, and widely used in food, pharmaceutical and cosmetic industry. The low concentration in plants and the difficulties in extraction restrict its large scale production. Saccharomyces cerevisiae can provide the monoterpene precursor, geranyl diphosphate (GPP) through its endogenous isoprenoid pathway. Therefore, it could be used as the host for monoterpene production. However, the weak metabolic flux through the isoprenoid pathway leads to the insufficient supply of GPP, and results in low monoterpene productivity. In order to increase the metabolic flux, we constructed the integrated expression plasmid pRS305-tHMG1 and free expression plasmid pYLIS-IDI1 to enhance the expression levels of isopentenyl diphosphate isomerase (IDI1) and a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase gene (tHMG1). The two plasmids were separately transformed into S. cerevisiae CEN.PK2-1C, resulting in strains LS01 and LS02. The plasmid pYLIS-IDI1 was further transformed into strain LS01, resulting in strain LS03. GC-MS analysis showed that the linalool concentration was increased by 1.3 times and reached (127.71 +/- 7.68) microg/L. In conclusion, enhancement of the supply of GPP precursors through the regulation of isoprenoid pathway could increase the linalool production in S. cerevisiae.