Marine bioactive compound dieckol induces apoptosis and inhibits the growth of human pancreatic cancer cells PANC-1.

Pancreatic cancer, which threatens the global population, is a very aggressive disease with an increased mortality rate. Regarding the types of cancer, pancreatic cancer is prone to display significant resistance to conventional therapy, therefore there 5-year survival rate is only 2% to 9%. Bioactive metabolites of marine algae such as polysaccharides, chitin, carternoids, and sterols possess immense pharmacological properties and tend to be promising alternatives for cancer treatment. Dieckol is one such polyphenolic bioactive compound extracted from brown algae Ecklonia cava, which is proven to possess antioxidant, anti-inflammatory, antibacterial, antidiabetic properties. Therefore in the present study, we analyzed the anticancer property of dieckol on PANC-1 pancreatic carcinoma cells. The cytotoxicity property of dieckol against PANC-1 cells was assessed with 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay, and cell morphological analysis. The generation of reactive oxygen species by dieckol on PANC-1 was analyzed with DCFH-DA staining and confirmed by quantifying antioxidants levels in untreated and dieckol-treated PANC-1 cells. The induction of apoptosis was further evaluated with different staining techniques such as Rhodamine 123 staining, acridine orange/ethidium bromide staining, DAPI staining, propidium iodide staining and was confirmed by estimating the protein expression of apoptotic genes, Bax and Bcl2. Cell adhesion assay and estimation of inflammatory cytokines were performed to detect the inhibitory effect of dieckol against cancer cell progression. It is further confirmed by analyzing cancer cell progression proteins, that is, proliferating cell nuclear antigen and cyclin D1 expressions in untreated and dieckol-treated PANC-1 cells. Our overall results authentically prove dieckol persuasively induces apoptosis and inhibits the progression of human pancreatic cancer cells in vitro, suggesting dieckol as a potent marine-based phytochemical to treat pancreatic cancer.

Coreactant-Free Dual Amplified Electrochemiluminescent Biosensor Based on Conjugated Polymer Dots for the Ultrasensitive Detection of MicroRNA.

Generally, electrochemiluminescence (ECL) assays are performed in the presence of a coreactant. The addition of the coreactant in the detection solution would make the ECL system lack sufficient stability. In the case of dissolved oxygen as the coreactant, the unknown concentration of dissolved O2 would result in an inevitable error and a lack of reproducibility in detection. A coreactant-free ECL assay could overcome the above shortcomings and thus is an ideal choice. In this work, a coreactant-free dual amplified ECL strategy was constructed for ultrasensitive detection of microRNA (miRNA). Here, target-catalyzed hairpin assembly and enzyme-triggered DNA walker recycling amplification were integrated to achieve dual signal amplification. Carboxyl-functionalized poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1'-3}-thiadiazole)] (PFBT-COOH) dots were used as luminophores, which displayed prominent ECL performance without adding any coreactants and removing the dissolved O2. As a result, the detection of miRNA was achieved, and the linear range was from 10 aM to 5 pM, and the detection limit was low to 3.3 aM. Meanwhile, the practicability of our biosensor was investigated by analyzing the expression of miRNA in cell lysates. The PFBT-COOH dots provided a great platform for constructing coreactant-free ECL biosensors and expanded the application of conjugated polymer dots in clinical analysis.

Screening potential reference genes for quantitative real-time PCR analysis in the oriental armyworm, Mythimna separata.

The oriental armyworm, Mythimna separata, is a major insect pest in China and other Asian countries. Unfortunately, suitable reference genes for quantitative real-time PCR (qRT-PCR) have not been previously identified in M. separata for normalizing target gene expression. In this study, we evaluated the expression stability of eight candidate genes (18S, ACT, EF1-α, GAPDH, RPS7, RPS13, RPL32 and TUB) in M. separata using the comparative ΔCt method, BestKeeper, Normfinder geNorm and ReFinder, a comprehensive software platform. The results indicated that the appropriate reference gene varied depending on the experimental conditions. We found that ACTIN, EF1-α and TUB were optimal for different developmental stages; TUB, RPS13 and EF1-α showed the most stable expresssion in different tissues; RPS13 and 18S were the best reference genes for monitoring expression under high temperature conditions; TUB, RPS13 and RPS7 exhibited the most stable expression under larval-crowding conditions; RPS7, EF1-α, RPL32 and GAPDH were the best for pesticide exposure experiments. This study provides tools for reliable normalization of qRT-PCR data and forms a foundation for functional studies of target gene expression in M. separata.

An electrochemiluminescent sensor based on functionalized conjugated polymer dots for the ultrasensitive detection of Cu2.

An ultrasensitive electrochemiluminescence (ECL) detection for Cu2+ was explored using the carboxyl functionalized poly(9,9-dioctylfluorenyl-2,7-diyl) (PS-COOH-co-PFO) dots as the signal label without adding any coreactant.

Mangiferin inhibits apoptosis and oxidative stress via BMP2/Smad-1 signaling in dexamethasone-induced MC3T3-E1 cells.

Mangiferin is a xanthone glucoside, which possesses antioxidant, antiviral, antitumor and anti-inflammatory functions, and is associated with gene regulation. However, it remains unknown whether mangiferin protects osteoblasts, such as the MC3T3-E1 cell line, against glucocorticoid-induced damage. In the present study, MC3T3-E1 cells were treated with dexamethasone (Dex), which is a well-known synthetic glucocorticoid, in order to establish a glucocorticoid-induced cell injury model. After Dex and/or mangiferin treatment, cell viability, apoptosis and reactive oxygen species (ROS) production was measured by Cell Counting kit-8 (CCK-8) and flow cytometry, respectively, and the concentration of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and macrophage colony-stimulating factor (M-CSF) was measured by ELISA. The expression of bone morphogenetic protein 2 (BMP2), phosphorylated­SMAD family member 1 (p-Smad-1), t-Smad-1, osterix (OSX), osteocalcin (OCN), osteoprotegerin (OPG), receptor activator of nuclear factor-κB (RANK), RANK ligand (RANKL), B­cell lymphoma 2 (Bcl-2) and Bcl­2­associated X protein (Bax) was measured by real-time PCR and/or western blot analysis. The results indicated that pretreatment of MC3T3-E1 cells with mangiferin for 3 h prior to exposure to Dex for 48 h significantly attenuated Dex-induced injury and inflammation, as demonstrated by increased cell viability, and decreases in apoptosis, ROS generation, and the secretion of TNF-α, IL-6 and M-CSF. In addition, pretreatment with mangiferin markedly reduced Dex-induced BMP2 and p­Smad-1 downregulation, and corrected the expression of differentiation­ and apoptosis­associated markers, including alkaline phosphatase, OSX, OCN, OPG, RANK, RANKL, Bcl-2 and Bax, which were altered by Dex treatment. Similar to the protective effects of mangiferin, overexpression of BMP2 suppressed not only Dex-induced cytotoxicity, but also ROS generation, and the secretion of TNF-α, IL-6 and M-CSF. In conclusion, the results of the present study are the first, to the best of our knowledge, to demonstrate that mangiferin protects MC3T3-E1 cells against Dex-induced apoptosis and oxidative stress by activating the BMP2/Smad-1 signaling pathway.

Roc curves of obesity indicators have a predictive value for children hypertension aged 7-17 years.

OBJECTIVE: The aim of the study is to examine the distribution of integrated covariate and its association with blood pressure (BP) among children in Anhui province, China, and assess the predictive value of integrated covariate to children hypertension. METHODS: A total of 2,828 subjects (1,588 male and 1,240 female) aged 7-17 years participated in this study. Height, weight, waistline, hipline and BP of all subjects were measured, obesity and overweight were defined by an international standard, specifying the measurement, the reference population, and the age and sex specific cut off points. High BP status was defined as systolic blood pressure (SBP) and/or diastolic blood pressure (DBP) ≥95th percentile for age and gender. RESULTS: Our results revealed that the prevalence of children hypertension was 11.03%, the SBP and DBP of obesity group were significantly higher than that of normal group. Anthropometric obesity indices such as body mass index (BMI) were positively correlated with SBP and DBP. Integrated covariate had a better performance than the single covariate in the receiver-operating characteristic (ROC) curve, the cut-off value; the sensitivity and the specificity of the integrated covariate were 0.112, 0.577, 0.683, respectively. CONCLUSION: Integrated covariate is a simple and effective anthropometric index to identify childhood hypertension.

Objetivo: El objetivo del estudio es examinar la distribución de las covariables integrado y su asociación con la presión arterial (PA) entre los niños en la provincia de Anhui, China, y evaluar el valor predictivo de covariables integrado a los niños la hipertensión. Métodos: Un total de 2.828 sujetos (1.588 macho y 1.240 hembra) de 7 a 17 años participaron en este estudio. Altura, peso, cintura, hipline y BP de todos los sujetos fueron medidos, la obesidad y el sobrepeso se han definido por una norma internacional, especificando la medición, la población de referencia, y la edad y sexo los puntos de corte.La condición de alta presión se define como la presión arterial sistólica (PAS) y/o presión arterial diastólica (PAD) ≥percentil para edad y sexo. Resultados: Nuestros resultados revelan que la prevalencia de niños la hipertensión fue 11,03%, el SBP y DBP de obesidad grupo fueron significativamente más alta que la de grupo normal. Los índices de obesidad, tales como el índice de masa corporal (IMC) se correlacionaron positivamente con SBP y DBP. Integrated covariable tuvo un mejor rendimiento que la covariable en la curva de características operativas del receptor (ROC), el valor de corte, la sensibilidad y la especificidad de las covariables fueron integrados 0,112, 0,577, 0,683, respectivamente. Conclusión: Integrado covariable es un simple y eficaz para identificar a la niñez índice antropométrico hipertensión.

ROC curves of obesity indicators have a predictive value for children hypertension aged 7-17 years / Curvas ROC de la obesidad en los indicadores tienen un valor predictivo para niños de entre 7 y 17 años hipertensión

Objective: The aim of the study is to examine the distribution of integrated covariate and its association with blood pressure (BP) among children in Anhui province, China, and assess the predictive value of integrated covariate to children hypertension. Methods: A total of 2,828 subjects (1,588 male and 1,240 female) aged 7-17 years participated in this study. Height, weight, waistline, hipline and BP of all subjects were measured, obesity and overweight were defined by an international standard, specifying the measurement, the reference population, and the age and sex specific cut off points. High BP status was defined as systolic blood pressure (SBP) and/or diastolic blood pressure (DBP) ≥ 95th percentile for age and gender. Results: Our results revealed that the prevalence of children hypertension was 11.03%, the SBP and DBP of obesity group were significantly higher than that of normal group. Anthropometric obesity indices such as body mass index (BMI) were positively correlated with SBP and DBP. Integrated covariate had a better performance than the single covariate in the receiver-operating characteristic (ROC) curve, the cut-off value; the sensitivity and the specificity of the integrated covariate were 0.112, 0.577, 0.683, respectively. Conclusion: Integrated covariate is a simple and effective anthropometric index to identify childhood hypertension (AU)

Objetivo: El objetivo del estudio es examinar la distribución de las covariables integrado y su asociación con la presión arterial (PA) entre los niños en la provincia de Anhui, China, y evaluar el valor predictivo de covariables integrado a los niños la hipertensión. Métodos: Un total de 2.828 sujetos (1.588 macho y 1.240 hembra) de 7 a 17 años participaron en este estudio. Altura, peso, cintura, hipline y BP de todos los sujetos fueron medidos, la obesidad y el sobrepeso se han definido por una norma internacional, especificando la medición, la población de referencia, y la edad y sexo los puntos de corte. La condición de alta presión se define como la presión arterial sistólica (PAS) y/o presión arterial diastólica (PAD) ≥ percentil para edad y sexo. Resultados: Nuestros resultados revelan que la prevalencia de niños la hipertensión fue 11,03%, el SBP y DBP de obesidad grupo fueron significativamente más alta que la de grupo normal. Los índices de obesidad, tales como el índice de masa corporal (IMC) se correlacionaron positivamente con SBP y DBP. Integrated covariable tuvo un mejor rendimiento que la covariable en la curva de características operativas del receptor (ROC), el valor de corte, la sensibilidad y la especificidad de las covariables fueron integrados 0,112, 0,577, 0,683, respectivamente. Conclusión: Integrado covariable es un simple y eficaz para identificar a la niñez índice antropométrico hipertensión (AU)

Urinary methylmalonic acid as an indicator of early vitamin B12 deficiency and its role in polyneuropathy in type 2 diabetes.

The rising incidence of diabetes and its negative impact on quality of life highlights the urgent need to develop biomarkers of early nerve damage. Measurement of total vitamin B12 has some limitations. We want to determine the levels of urinary methylmalonic acid and its relationships with serum vitamin B12 and polyneuropathy. The 176 Chinese patients with Type 2 diabetes mellitus were divided into 3 groups according to the levels of vitamin B12. A gas chromatography mass spectrometric technique was used to determine blood methylmalonic acid and urinary methylmalonic acid. The diagnosis of distal diabetic polyneuropathy was based on the determination of bilateral limb sensory and motor nerve conduction velocity and amplitude with electromyogram. Multiple regression analysis revealed that urinary methylmalonic acid/creatinine, blood methylmalonic acid, and so forth were variables that influenced diabetic polyneuropathy significantly. Nerve sensory conduction velocity and nerve amplitude in the group of urinary methylmalonic acid/creatinine >3.5 mmol/mol decreased significantly. Superficial peroneal nerve sensory and motor conduction velocity and ulnar nerve compound motor active potential amplitude were inversely correlated with urinary methylmalonic acid/creatinine. Urinary methylmalonic acid correlates with serum vitamin B12 levels in person with diabetes and is a sensitive marker of early polyneuropathy.

Dipeptidyl peptidase-IV is a potential molecular biomarker in diabetic kidney disease.

The present study was designed to identify the changes in microvesicle-dipeptidyl peptidase-IV (DPP IV) levels in human urine and serum, and to determine whether there were correlations with the severity of diabetic kidney disease (DKD). A total of 127 patients with type 2 diabetes mellitus (T2DM) were divided into three groups according to the urinary albumin/ creatinine ratio (UACR): microalbuminuria group (n = 50); macroalbuminuria group (n = 34) and normoalbuminuria group (n = 43), and 34 age- and sex-matched non-diabetic healthy subjects were selected as controls. Microvesicle-bound DPP IV and free urinary DPP IV were separated by a filtra-centrifugation method. The total microvesicles were captured by a specific monoclonal antibody, AD-1. DPP IV activity was determined by measuring the cleavage of chromogenic free 4-nitroaniline from Gly-Pro-p-nitroanilide at 405 nm with an ELISA plate reader. DPP IV protein levels were determined by ELISA and Western blot. Our results showed that the microvesicle-bound type was the major form of DPP IV in urine; the urinary microvesicle-DPP IV excretion of each T2DM group was significantly higher compared with controls. The urinary microvesicle-DPP IV level was positively correlated with UACR in patients with T2DM. These findings suggest that the urinary level of microvesicle-bound DPP IV is associated with the severity of DKD.

The construction of glucose biosensor based on platinum nanoclusters-multiwalled carbon nanotubes nanocomposites.

One-step synthesis method was proposed to obtain the nanocomposites of platinum nanoclusters and multiwalled carbon nanotubes (PtNCs-MWNTs), which were used as a novel immobilization matrix for the enzyme to fabricate glucose biosensor. The fabrication process of the biosensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, atomic force microscopy and scanning electron microscope. Due to the favorable characteristic of PtNCs-MWNTs nanocomposites, the biosensor exhibited good characteristics, such as wide linear range (3.0 µM-12.1 mM), low detection limit (1.0 µM), high sensitivity (12.8 µA mM⁻¹), rapid response time (within 6 s). The apparent Michaelis-Menten constant (K(app)(m)) is 2.1 mM. The performance of the resulting biosensor is more prominent than that of most of the reported glucose biosensors. Furthermore, it was demonstrated that this biosensor can be used for the assay of glucose in human serum samples.

[Effects of electroacupuncture at "Zusanli" (ST 36) on Pharmacokinetics of paracetamol in rates].

OBJECTIVE: To investigate influence of electroacupuncture at "Zusanli" (ST 36) on pharmacokinetics after oral administration of Paracetamol in rats and explore the effect and mechanism of acupuncture combined with medicinal drugs. METHODS: Forty-eight SD rats, in half respectively male and female, were randomly divided into 6 groups: Paracetamol groups of low, moderate, and high doses (oral dose of Paracetamol 300, 600 or 1200 mg/kg, respectively), and electroacupuncture plus Paracetamol groups of low, moderate, and high doses (electroacupuncture at "Zusanli" (ST 36) acupoint for 20 min after oral dose of Paracetamol 300, 600 or 1 200 mg/kg, respectively). Serum specimens of rats in each group were obtained at different time points to determine the concentrations of Paracetamol by RP-HPLC after oral administration of Paracetamol at different dosage, and the pharmacokinetic parameters were calculated by software 3P87, so as to observe the influence of electroacupuncture on absorption and metabolism of Paracetamol. RESULTS: Except in electroacupuncture plus Paracetamol groups of high doses group showing a non-linear dynamics model, the pharmacokinetics parameters of Paracetamol in the other fives groups were all fitted to first order grade absorption of two-compartment open model. Compared with the Paracetamol groups of low and moderate doses, the time to peak concentration (T(max)) was advanced (P < 0.01), the peak values of plasma concentration (C(max)) were significantly higher (P < 0.01), and the absorption half-life (T1/2ka) speeded up apparently (P < 0.01), but the elimination half-life (T1/2ke) remained basically unchanged (P > 0.05), and the area under the curve (AUC) increased significantly (P < 0.01), and plasma clearance (Cl(s)) was lower (P < 0.01) in the electroacupuncture plus Paracetamol groups of low and moderate doses. Compared with the Paracetamol group of high doses, the AUC also increased significantly (P < 0.01) and the mean residence time (MRT) postponed relatively (P < 0.05) in the electroacupuncture plus Paracetamol group of high doses. CONCLUSION: It indicates that clinical consideration must be given to the drug dosage and the possible influence of electroacupuncture on the metabolism of some drugs in order to avoid and reduce adverse reactions.

[Segmental resection of the liver by Glissonean pedicle transection for primary liver cancer].

OBJECTIVE: To study the clinical effect of segmental resection of the liver using Glissonean pedicle transection for primary liver cancer. METHODS: The clinical data of 55 primary liver cancer patients admitted from January 2006 to October 2008 were analyzed retrospectively. Twenty-five of the patients underwent segmental resection of the liver by Glissonean pedicle transection (group A), and 30 underwent routine hepatectomy (group B). The positivity rate of the resection margin, micrometastasis in the hepatic parenchyma surrounding the lesions and postoperative recurrence rates were investigated. RESULTS: The positivity rate of the resection margin was 4.0% in group A, significantly lower than that of group B. The number of histological micrometastasis was significantly higher in group A than in group B (16 vs 8). The median distance of histological micrometastasis was 6.8 mm (2.7-25.6 mm) in group A and 4.2 mm (2.4-9.0 mm) in group B. The one-year recurrence rate was significantly lower in group A than in group B (16% vs 26.7%). CONCLUSION: Glissonean pedicle transection for segmental liver resection is a simpler procedure than routine hepatectomy for primary liver cancer and can reduce the number of histological micrometastasis and recurrence rate.

[The relationship between resistin and the peripheral neuropathy in type 2 diabetes].

OBJECTIVE: To investigate the relationship between the serum resistin concentration and type 2 DM with peripheral neuropathy. METHODS: Ninety-six patients of type 2 diabetes mellitus (DM) among which 52 were with peripheral neuropathy (DM + PN) and 34 healthy persons undergoing routine physical examination as controls were studied. Body weight, waist circumference, blood pressure, body mass index (BMI), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), and beta-cell action measured by the homeostasis model assessment (HOMA-beta), and blood lipids were measured. ELISA was used to detect the serum resistin, Endothelins (ET), and nitrogen oxide (NO). RESULTS: The serum resistin level of the DM + PN patients was (26 +/- 9) ng/ml, significantly higher than that of the DM patients [(20 +/- 6) ng/ml, P < 0.05]. Th serum resistin level was positively correlated with FINS, Homa-IR, systolic blood pressure, diastolic blood pressure, and ET (all P < 0.05) and significantly negatively correlated with HOMA-beta and NO (both P < 0.05). CONCLUSION: Resistin may play a role in the pathogenesis of type 2 DM and diabetic peripheral neuropathy.