Apelin-13 attenuates high glucose-induced calcification of MOVAS cells by regulating MAPKs and PI3K/AKT pathways and ROS-mediated signals.

Vascular calcification (VC) is an inducement of many cardiovascular diseases. Clinic evidences have confirmed that diabetes was the independent risk factor for VC, and the mechanism has not been well explored. Apelin as a ligand molecule is widely found in the cardiovascular system and showed potential in inhibiting VC, but the inhibitory effect and mechanism of apelin-13 against high glucose-induced VC have not been investigated yet. Herein, apelin-13 was employed to inhibit high glucose-induced VC in mouse aortic vascular smooth muscle cells (MOVAS), and the underlying mechanism was explored. The results showed that apelin-13 significantly inhibited high glucose-induced cells proliferation, migration and invasion of MOVAS cells. Apelin-13 also effectively attenuated high glucose-induced calcification by inhibiting alkaline phosphatase (ALP) activity and expression. Further investigation revealed that apelin-13 dramatically suppressed high glucose-induced DNA damage through inhibiting reactive oxide species (ROS) generation. Moreover, apelin-13 also effectively improved high glucose-induced dysfunction of MAPKs and PI3K/AKT. Inhibition of ERK by inhibitor (U0126) significantly blocked high glucose-induced calcification, which further confirmed the significance of MAPKs. Taken together, these results suggested that apelin-13 had the potential to attenuate high glucose-induced calcification of MOVAS cells by inhibiting ROS-mediated DNA damage and regulating MAPKs and PI3K/AKT pathways. Our findings validated the strategy of using apelin-13 maybe a novel way in treating high glucose-mediated VC.

Changes of hippocampus proteomic profiles after blueberry extracts supplementation in APP/PS1 transgenic mice.

Objective: To examine protein changes in the hippocampus of APP/PS1 transgenic mice after blueberry extracts (BB) intervention.Methods: Eight APP/PS1 transgenic mice were randomly assigned to Alzheimer's disease (AD)+BB group (n=4) and AD+control group (n=4). After a 16-week treatment, 2-DE and MALDI-TOF-MS were used to compare the proteomic profiles of the hippocampus in the two groups and Western blot was used to confirm the important differentially expressed proteins.Results: Twelve proteins were differentially expressed between the two groups. Nine of them were identified. Cytochrome b-c1 complex subunit 6, beta-actin, dynamin 1, and heat shock cognate 71 were up-regulated in AD+BB group, while a-enolase, stress-induced-phosphoprotein 1, malate dehydrogenase (MDH), MDH 1, and T-complex protein 1 subunit beta were down-regulated, respectively. Importantly, some of the identified proteins (e.g. dynamin 1) are known to be involved in cognitive impairment. Western blot analysis of hippocampus dynamin 1 expression confirmed the proteomic findings.Conclusions: The consumption of BB modulates the expression of proteins that are linked to the improvements of cognitive dysfunction in hippocampus of APP/PS1 transgenic mice.

Cyanidin-3-O-galactoside and blueberry extracts supplementation improves spatial memory and regulates hippocampal ERK expression in senescence-accelerated mice.

OBJECTIVE: To investigate whether the antioxidation and the regulation on the Extracellular Regulated Protein Kinases (ERK) signaling pathway are involved in the protective effects of blueberry on central nervous system. METHODS: 30 Senescence-accelerated mice prone 8 (SAMP8) mice were divided into three groups and treated with normal diet, blueberry extracts (200 mg/kg•bw/day) and cyaniding-3-O-galactoside (Cy-3-GAL) (50 mg/kg•bw/day) from blueberry for 8 weeks. 10 SAMR1 mice were set as control group. The capacity of spatial memory was assessed by Passive avoidance task and Morris water maze. Histological analyses on hippocampus were completed. Malondialdehyde (MDA) levels, Superoxide Dismutase (SOD) activity and the expression of ERK were detected. RESULTS: Both Cy-3-GAL and blueberry extracts were shown effective functions to relieve cellular injury, improve hippocampal neurons survival and inhibit the pyramidal cell layer damage. Cy-3-GAL and blueberry extracts also increased SOD activity and reduced MDA content in brain tissues and plasma, and increased hippocampal phosphorylated ERK (p-ERK) expression in SAMP8 mice. Further more, the passive avoidance task test showed that both the latency time and the number of errors were improved by Cy-3-GAL treatment, and the Morris Water Maze test showed significant decreases of latency were detected by Cy-3-GAL and blueberry extracts treatment on day 4. CONCLUSION: Blueberry extracts may reverse the declines of cognitive and behavioral function in the ageing process through several pathways, including enhancing the capacity of antioxidation, altering stress signaling. Cy-3-GAL may be an important active ingredient for these biological effects.

Depletion of intracellular zinc induced apoptosis in cultured hippocampal neurons through Raf/MEK/ERK pathways.

An experiment was performed to observe the changes in Raf-1 kinase/mitogen-activated protein kinase ERK (MEK)/extracellular signal-regulated kinase (ERK) signaling pathways in cultured hippocampal neurons and its correlation with neurons apoptosis induced by intracellular zinc depletion. Cultured hippocampal neurons were exposed to a cell membrane-permeant zinc chelator TPEN (2 µM), and to TPEN plus zinc sulfate (5 µM) for 24 h. Cultures were then processed to detect neuronal viability by the methyl thiazolyl tetrazolium assay, while apoptosis rate was simultaneously observed by the flow cytometric analysis. Caspase-3, Raf-1, pMEK, pERK1/2, and pCREB protein levels were examined by Western blot assays. The viability in TPEN-incubated neurons was notably decreased, apoptosis rate and expression of caspase-3 significantly increased compared to untreated controls. The significant down-regulation of Raf/MEK/ERK signaling pathway and expression of pCREB were decreased in TPEN-treated neurons. Co-addition of zinc almost completely reversed TPEN-induced alterations described. The results demonstrated zinc-modulated apoptosis and the expression of Raf/MEK/ERK at the protein level in hippocampal neurons. It is possible that zinc depletion-induced apoptosis in cultured hippocampal neurons may be relevant to the changes of Raf/MEK/ERK signaling pathway.

Effects of intracellular zinc depletion on the expression of VDAC in cultured hippocampal neurons.

An experiment was conducted to investigate whether intracellular zinc depletion can actually change expression of voltage-dependent anion channel 1 (VDAC1) and VPAC2 in cultured hippocampal neurons as well as their significance. Hippocampal neurons were obtained by primary culture from hippocampus of newborn Wistar rats. Cultured hippocampal neurons were exposed to a cell membrane-permeable zinc chelator N,N,N',N'-tetrakis (2-pyridyl methyl) ethylenediamine (TPEN) (2 µM), and to TPEN plus zinc sulfate (5 µM) for 1 or 24 hours. Cultures were then processed to detect neuronal injury by lactate dehydrogenase (LDH) assay, intracellular Ca(2+) with the fluorescent probe fluo-3/AM, reactive oxygen species (ROS) generation using 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay, nuclear morphology by Hoechst 33342, VDAC1, and VDAC2 protein levels by western blot, and VDAC1 and VDAC2 mRNA levels by RT-PCR. The results demonstrated that exposure of hippocampal neurons to TPEN (2 µM) for 24 hours induced notably neuronal injury, significantly increased the number of apoptotic nuclei, up-regulated the expression of VDAC1 protein level and down-regulated the expression of VDAC2 protein level. Significant down-regulation of mRNA levels for VDAC1 and VDAC2 were observed in TPEN-treated neurons. Co-addition of zinc almost completely reversed TPEN-induced neuronal injury and above alterations in VDAC1 and VDAC2 protein levels and mRNA levels. Present results implicate a possibility that up-regulation of VDAC1 and down-regulation of VDAC2 may participate in hippocampal neuron injury induced by zinc deficiency.

Expression of dopamine receptor D2 in pulmonary carcinoma cells and effects of dopamine on the cell apoptosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of dopamine receptor D2 in different pulmonary carcinoma cells and investigate the effect of dopamine in inducing apoptosis of A549 cells.</p><p><b>METHODS</b>Western blotting and RT-PCR were employed to detect the expression of dopamine receptor D2 in different pulmonary carcinoma cells (95D, H460, GLC-82, A549 and H446 cells). The apoptosis of A549 cells after a 6-hour exposure to 0.02%, 0.04%, 0.08% and 0.1% dopamine was analyzed by flow cytometry. The apoptosis-inducing effect of dopamine in vivo was also tested by intratumoral injection of 1% dopamine in 2 BALB/c-nu mice bearing A549 tumor xenograft using flow cytometry.</p><p><b>RESULTS</b>The presence of dopamine receptor D2 expression was detected by Western blotting and RT-PCR in 95D, H460, GLC-82, A549 and H446 cells. Flow cytometry detected obvious apoptosis of A549 cells following dopamine exposure in vitro in positive correlation to dopamine concentration. In the tumor-bearing mice, dopamine also showed an obvious apoptosis-inducing effect on A549 cells.</p><p><b>CONCLUSION</b>Dopamine receptor D2 exists extensively in different pulmonary carcinoma cells. Dopamine may promote the apoptosis of pulmonary carcinoma cells through dopamine receptor D2.</p>

Utility of squamous cell carcinoma antigen, carcinoembryonic antigen, Cyfra 21-1 and neuron specific enolase in lung cancer diagnosis: a prospective study from China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Early detection and diagnosis is urgent for the sake of effective treatment strategy for lung cancer. However, a convenient, economical and relatively precise method is not available. We here report a prospective study to find the possible value of the combined use of four popular tumor markers in the early diagnosis of lung cancer among patients with suspicious nodules in the lung.</p><p><b>METHODS</b>Six hundred and sixty inpatients with suspicious nodules in the lung were divided into a lung cancer group and a benign pulmonary tumor group according to post-operative histological examinations. Serum levels of four tumor markers including squamous cell carcinoma antigen (SCC), carcinoembryonic antigen (CEA), Cyfra 21-1 and neuron specific enolase (NSE) were assayed for each patient. Receiver operating characteristic (ROC) curves were constructed for each tumor marker. The power of lung cancer diagnosis of each tumor marker, as well as a combination of them were analyzed and compared.</p><p><b>RESULTS</b>The serum levels (median, range) of SCC, CEA, Cyfra 21-1 and NSE were 0.44 (0.01 - 35.70) ng/ml, 2.49 (0.30 - 26.78) ng/ml, 2.30 (0.82 - 73.33) ng/ml and 10.54 (0.10 - 56.41) ng/ml respectively in lung cancer group, and were 0.32 (0.01 - 0.90) ng/ml, 1.60 (0.20 - 8.93) ng/ml, 1.41 (0.72 - 4.82) ng/ml and 9.36 (6.56 - 24.24) ng/ml respectively in the benign pulmonary tumor group. The difference in each tumor marker between the two groups was significant (P < 0.05). The ROCs of SCC, CEA, Cyfra 21-1 and NSE were 0.702 (95%CI, 0.654 - 0.751), 0.611 (95%CI, 0.563 - 0.659), 0.650 (95%CI, 0.601 - 0.700) and 0.598 (95%CI, 0.542 - 0.654) respectively, indicating very low power of these four tumor markers. When a combination of SCC, CEA, Cyfra 21-1 and NSE were employed, the diagnosis power was strengthened.</p><p><b>CONCLUSION</b>SCC, CEA, Cyfra 21-1 and NSE are valuable in the early diagnosis of lung cancer among suspicious nodules in the lung, especially when they were assayed together for one patient.</p>

Polymorphism of MTHFR C677T, serum vitamin levels and cognition in subjects with hyperhomocysteinemia in China.

Relationships between hyperhomocysteinemia (HHE) and neurodegenerative diseases have been widely studied. However, the impact of serum total homocysteine (tHcy) levels on cognitive function has not been confirmed. C677T polymorphisms in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene have impacts on tHcy level; it is suspected to influence cognitive function, but only few investigations have assessed its effects on non-dementia adults and the results have been controversial. Moreover, there is no report about Chinese subjects. In the present study, we determined C677T/MTHFR genotype, serum tHcy concentration and cognition in 182 nondemented subjects aged 55-88 years to probe the associations between MTHFRC677T mutation, increased tHcy levels and decreased cognitive function in a northern city in China. A serum tHcy level > or = 16 micromol/l was deemed HHE. Cognitive function was assessed by the Mini Mental State Examination (MMSE) and Basic Cognitive Aptitude Tests (BCAT). Results showed that: (i) subjects with the T allele had higher serum tHcy levels than those without, especially in lower folate status; (ii) T allele and CT/TT genotype frequencies in subjects with HHE were higher than in non-HHE subjects (P < 0.05); and (iii) serum tHcy level was inversely related to total BCAT score (P < 0.05) but MTHFR677 C to T polymorphism had no association with it. Our results confirmed that the MTHFR 677 C to T mutation, especially in lower serum folate concentration status, results in the increase of serum tHcy levels which is bad for cognitive function and indicates that higher serum folate level is of benefit in keeping lower serum tHcy level and better cognitive function. The results provide some valuable clues for individualized nutrition intervention of HHE and cognition decline in the middle-aged and the elderly.

[Cytoprotective effect of blueberry extracts against oxidative damage of rat hippocampal neurons induced by H2O2].

OBJECTIVE: To investigate the protective effect of blueberry extract (BE, 25% anthocyanins) against oxidative damage in primary cultures of rat hippocampal neurons induced by H2O2. METHODS: Rat hippocampal neurons were randomly assigned to control group, H2O2 group and BE pretreatment groups, BE at six different doses (0.01, 0.1, 1.0, 10.0, 20.0 and 40 microg/ml) and then exposed to 50 micromol/L H2O2 for twenty-four hours. To selecte the most fittest concentration of BE by testing viability of neurons and activity of LDH. Then MDA concentration, SOD activity and neuronal apoptosis were(checked) measured. RESULTS: (1) Compared with H2O2 group, the hippocampal cell viabilities in the 0.1, 1.0 and 10 microg/ml BE groups were significantly increased from 57.44% to 78.42%, 87.71% and 72.40% separately. The activity of LDH in BE groups at varied concentrations (0.1, 1.0 and 10 microg/ml) was significiantly lower than that in H2O2 group. It was found that 1 microg/ml BE had the furthest protective effect against oxidative damage in primary cultures of rat hippocampal neurons induced by H2O2. (2) The concentration of MDA and the rate of neuronal apoptosis of BE group (1 microg/ml) were much lower than H2O2 group, while SOD activity was much higher. CONCLUSION: Proper dose of BE has remarkable protective effect against oxidative stress in primary cultures of rat hippocampal neurons induced by H2O2, the mechanism may be related to decreasing the neuronal apoptosis and enhancing the antioxidation of hippocampal neurons.

[Study on triterpenes from of Ligularia xanthotricha].

OBJECTIVE: To investigate the chemical constituents of Ligularia xanthotricha. METHOD: Silica gel column chromatography and preparative TLC were employed for the isolation and purification. The structures were identified on the basis of spectral data (IR, EI-MS, 1H-NMR, 13C-NMR, DEPT) and chemical evidence. RESULT: Seven compounds were isolated and identified as follows: lupeol (1), lupeol palmitate (2), 3, 28-dihydroxyl-lupeol (3), betulinic acid (4), taraxasterol (5), taraxasteryl palmitat (6) and taraxasteryl acetate(7). CONCLUSION: All the compounds were isolated from this plant for the first time.

[Studies on glycosides from Gentiana veitchiorum].

OBJECTIVE: To study the chemical constituents of Gentiana veitchiorum. METHOD: The chemical constituents were isolated by chromatography and identified by spectral data. RESULT: Five glycosides, loganic acid (1), gentiopicroside (2), isoorientin 3'-methyl ether (3), isovitexin (4), isoorientin (5) were isolated and identified. CONCLUSION: Compounds 1-5 were isolated from this plant for the first time.

Research on application of modified polyethylene glycol hydrogels in the construction of tissue engineered heart valve / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the effect of the polyethylene glycol (PEG)-hydrogels to enhance the seeding-cells adhesion to the biomaterial scaffolds.</p><p><b>METHODS</b>Sixteen porcine aortic valves were decellularized with Triton X-100 and trypsin, then divided into A and B group, eight in each group. Group A: the donor goat's autologous bone marrow mesenchymal stem cells (BMSCs) Selected as the seeding-cells were encapsulated into the modified PEG-hydrogels to complete the process of the cells attaching to the acellular porcine aortic valves. Non-PEG but reservation of BMSCs was modified in Group B. After static culture for 7 d, the mono semilunar tissue engineering heart valve (TEHV) were implanted respectively into each donor goat's abdominal aortas. Gross and histology examination, ultrasonic scanning, electron microscopy observation and biomechanics detection were performed at 16 weeks after operation. The 8 native goat aortic valves from the donor goats were selected at the same time as control group (Group C).</p><p><b>RESULTS</b>There were much more improvements compared Group A to Group B (P < 0.05) in tensile strength [(12.9 +/- 1.3) MPa vs. (8.8 +/- 0.4) MPa], ratio of re-endothelial (84.6% vs. 14.8%) and mural thrombosis (0/8 vs. 8/8). The data illustrated the critical importance of BMSCs differentiation to endothelial and myofibroblast for remodeling into native tissue in microenvironment in vivo.</p><p><b>CONCLUSIONS</b>It is feasible to reconstruct TEHV efficiently by combining modified PEG-hydrogels with acellular biomaterial scaffold and autologous MSCs cells. It can improve the integration of the seeding-cells and scaffold. It can also protect the growth and differentiation of the BMSCs in the systemic circulation effectively.</p>

Serum transforming growth factor-beta1 level reflects disease status in patients with esophageal carcinoma after radiotherapy.

AIM: To evaluate the relationship between changes in serum transforming growth factor beta1 (TGFbeta1) level and curative effect of radiotherapy (RT) in patients with esophageal carcinoma. METHODS: Ninety patients with histologically confirmed esophageal carcinoma were enrolled. Serum samples for TGFbeta1 analysis were obtained before and at the end of RT. An enzyme-linked immunosorbent assay was used to measure serum TGFbeta1 level. Multivariate analysis was performed to investigate the relationship between disease status and changes in serum TGFbeta1 level. RESULTS: Serum TGFbeta1 level in patients with esophageal carcinoma before RT was significantly higher than that in healthy controls (P < 0.001). At the end of RT, serum TGFbeta1 level was decreased in 67.82% (59/87) of the patients. The overall survival rate at 1, 3 and 5 years was 48.28% (42/87), 19.54% (17/87) and 12.64% (11/87), respectively. Main causes of death were local failure and regional lymph node metastasis. In patients whose serum TGFbeta1 level decreased after RT, the survival rate at 1, 3 and 5 years was 61.02% (36/59), 28.81% (17/59) and 18.64% (11/59), respectively. The survival rate at 1 year was 17.86% (5/28) in patients whose serum TGFbeta1 level increased after RT, and all died within 18 mo (P < 0.01). CONCLUSION: Serum TGFbeta1 level may be a useful marker for monitoring disease status after RT in patients with esophageal carcinoma.

[Fluorescence spectrum analysis system for protoporphyrin IX in serum based on wavelet transform].

Protoporphyrin IX is an important kind of organic compound for vital movement, and can be used as the sign of tumour blood. Human protoporphyrin IX content in serum is very low, and affected by various factors. The serum fluorescence spectrum analysis system based on wavelet transform was used to discriminated the protoporphyrin IX weak signals. The protoporphyrin IX fluorescence spectrum was obtained by a multi-function spectrum measuring system, and decomposed several times by wavelet transform to distinguish the noise and spectrum signals. The fluorescence spectrum can be divided into corresponding discrete approximations signals (a1-a6) and discrete details signals (d1-d6) by six times of decomposition, showing the signal frequency decreasing with decomposition times increasing and the protoporphyrin IX fluorescence character peak appears here. The weak signals were discriminated and the exactly component and quantity can be acquired for further analysis. So it can be analysed quantitatively. The researches in the present paper provide the potential application in the diagnosis of incipient tumous using the serum fluorescence spectrum

Two new polyporusterones from Polyorus umbellatus.

Two new polyporusterones named as porusterone I and polyprosterone II were isolated from polyourus umbellatus. Their structures have been established on the basis of spectroscopic analysis.