Naringin attenuates acute myocardial ischemia-reperfusion injury via miR126/GSK-3ß/ß-catenin signaling pathway

Introduction: Myocardial ischemia-reperfusion (I/R) injury is one of the mechanisms contributing to the high mortality rate of acute myocardial infarction. Purpose: This study intended to study the role of naringin in cardiac I/R injury. Methods: AC16 cells (human cardiomyocyte cell line) were subjected to oxygen-glucose deprivation/recovery (OGD/R) treatment and/or naringin pretreatment. Then, the apoptosis was examined by flow cytometry and Western blotting. The concentration of IL-6, IL-8 and TNF-α was measured by enzyme-linked immunosorbent assay (ELISA) kits. How naringin influenced microRNA expression was examined by microarrays and quantitative real-time polymerase chain reaction (qRT-PCR). Dual luciferase reporter assay was employed to evaluate the interaction between miR-126 and GSK-3ß. The GSK-3ß/ß-catenin signaling pathway was examined by Western blotting. Finally, rat myocardial I/R model was created to examine the effects of naringin in vivo. Results: Naringin pretreatment significantly decreased the cytokine release and apoptosis of cardiomyocytes exposed to OGD/R. Bioinformatical analysis revealed that naringin upregulated miR-126 expression considerably. Also, it was found that miR-126 can bind GSK-3ß and downregulate its expression, suggesting that naringin could decrease GSK-3ß activity. Next, we discovered that naringin increased ß-catenin activity in cardiomyocytes treated with OGD/R by inhibiting GSK-3ß expression. Our animal experiments showed that naringin pre-treatment or miR-126 agomir alleviated myocardial I/R. Conclusions: Naringin preconditioning can reduce myocardial I/R injury via regulating miR-126/GSK-3ß/ß-catenin signaling pathway, and this chemical can be used to treat acute myocardial infarction.

Expression and clinical significance of hsa-miRNA-200c in patients with liver metastasis of epithelial ovarian cancer / 中国实用妇科与产科杂志

OBJECTIVE: To investigate the expressions of hsa-miRNA-200 c and its relationship with metastasis of EOC.METHODS: The expression of hsa-miRNA-200 c was detected by Stem-loop Real-time Quantitative PCR(TaqMan probe method)in 73 cases of EOC,30 cases of benign ovarian epithelial tumors and 30 cases of normal ovarian tissues,which were collected in gynecological operations from Guangdong General Hospital from October 2010 to May 2011.Meantime,the clinical pathologic features data were analyzed.The assessment of the correlation between hsa-miRNA-200 c and clinicopathological features,and the hierarchical analysis of hsa-miRNA-200 c level in 73 cases of ovarian epithelial cancer was further undertaken(Among 73 cases,13 patients suffered liver metastasis and 60 patients had non-liver metastasis).Overexpression or knockdown of hsa-miRNA-200 c,ovarian cancer cell invasion and migration abilitywas detected.RESULTS: The expression of miRNA-200 c in the EOC tissues was 382.18±15.22,which was significantly higher than that in the benign ovarian epithelial tumors(35.61 ± 1.42)and normal ovarian tissues(4.43 ±2.23)(P0.05).The expressions of miRNA-200 c were low in EOC with late clin-ical FIGO stage(670.91±16.88 vs. 129.52±33.3,P0.05).The transwell cabinet invasion experiment showed the expression of miRNA-200 c was negatively correlated with the invasion capability of ovarian cancer cells.CONCLUSION: MiRNA-200 c is likely to play a double regulation role in the development of EOC,whose low-expression has been associated with late EOC,lymph node metastasis,liver metastasis,and poor prognosis.

The dynamic expression changes of myocardium p-p38MAPK, NF-κB and COX-2 in rats after an exhausted exercise / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the dynamic expression changes of p38 mitogen-activated protein kinase (p38MAPK), nucler facter kappa B (NF-κB) and cyclooxygenase-2 (COX-2) in myocardial tissue after an exhausted exercise and study the impact of p38MAPK, NF-κB and COX-2 on its myocardial damage.</p><p><b>METHODS</b>Sixty Wister male rats were randomly divided into the control group (n = 10) and the exhausted exercise group (n = 50). Then the exhausted exercise group was further divided into 5 subgroups, namely 0 h, 3 h, 6 h, 12 h, 24 h after an exhausted exercise (n = 10). The myocardial injury animal model was set up by using an exhausted swimming exercise and the expression of p-p38MAPK, NF-κB and COX-2 were examined by Western blot.</p><p><b>RESULTS</b>Compared with the control group, the expression of p-p38MAPK were increased significantly (P < 0.01) in all the groups and the 3 h group was the highest( P < 0.01); The expression of NF-κB were increased significantly (P < 0.05) in all the groups but 0 h P > 0.05) and the 6h group was increased significantly compared with the other groups( P < 0.05); The expression of COX-2 were increased significantly( P < 0.05) in all the groups but 0 h and the 24 h groups was increased significantly compared with the other groups(P < 0.05).</p><p><b>CONCLUSION</b>p38MAPK was activated in an acute exhausted exercise, p-p38MAPK may play an important role in modulating NF-κB and COX-2 expression and mediating the exhausted exercise induced myocardial damage.</p>

Changes of c-fos, malondialdehyde and lactate in brain tissue after global cerebral ischemia under different brain temperatures / 华中科技大学学报（医学）（英德文版）

Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic (37-38°C) ischemia, mild hypothermic (31-32°C) ischemia, hyperthermic (41-42°C) ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20-min ischemia. The expression of c-fos protein and the levels of malondialdehyde (MDA) and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-fos protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion following 20-min ischemia as compared with the sham-operated group (P<0.01). The levels of MDA and lactate in mild hypothermic group were significantly lower than those in normothermic group (P<0.01). It is suggested that brain temperature influences the translation of the immunoreactive protein product of c-fos after global cerebral ischemia, and MDA and lactate are also affected by hypothermia and hyperthermia.

Changes of c-fos, malondialdehyde and lactate in brain tissue after global cerebral ischemia under different brain temperatures / 华中科技大学学报（医学）（英德文版）

Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic (37-38°C) ischemia, mild hypothermic (31-32°C) ischemia, hyperthermic (41-42°C) ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20-min ischemia. The expression of c-fos protein and the levels of malondialdehyde (MDA) and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-fos protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion following 20-min ischemia as compared with the sham-operated group (P<0.01). The levels of MDA and lactate in mild hypothermic group were significantly lower than those in normothermic group (P<0.01). It is suggested that brain temperature influences the translation of the immunoreactive protein product of c-fos after global cerebral ischemia, and MDA and lactate are also affected by hypothermia and hyperthermia.

The expression of interferon-lambda1 in CHO cell / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which linked the enhancer SP163 with interferon lambda1. Then express the interferon lambda1 in CHO (dhfr-) cells.</p><p><b>METHODS</b>Using PCR method to introduce the restriction enzyme sites and through the fusion PCR binding the enhancer with the interferon Lambda1. After sequenced, lambda1 and SP163-lambda1 was inserted into PCI-dhfr forming the expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which was constructed successfully confirming by sequencing. Then the expressing vectors were transfected into CHO (dhfr-) cells using liposome transfection method and interferon lambda1 protein was assayed with indirect immunofluorescence and Western Blot. Using cytopathic effect inhibition evaluated the antiviral activity of interferon lambda1.</p><p><b>RESULTS</b>Successfully constructing the eukaryotic expression vectors of interferon lambda and the vectors could express interferon lambda1. The result of immunofluorescence showed the enhancer developed the expression of interferon lambda1. Detecting the interferon lambda1 in CHO (dhfr-) cells after transfecting 48 hour using Western Blot. The cytopathic effect inhibition showed the expressed interferon lambda1 has the antiviral activity.</p><p><b>CONCLUSION</b>Successfully expressed the interferon lambda1 in CHO (dhfr-) cells and the protein possesses antiviral activity, which may supply a valuable basis for building the stable cell line of interferon lambda1.</p>

Preparation of the monoclonal antibody against human Bocavirus VP2 protein / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express and purify HBoV VP2 protein, and the monoclonal antibody against HBoV VP2 protein was prepared with hybridoma technique.</p><p><b>METHODS</b>The HBoV VP2 cloned into vector pET-30a was expressed in E. coil. After purified by immobilized metal affinity chromatography, the BALB/c mouse was immunized with purified protein as antigen. The positive hybridoma cells were screened with hybridoma technique and ELISA assay. Isotype and titer of the monoclonal antibody were detected.</p><p><b>RESULTS</b>The recombinant HBoV VP2 protein was expressed and purified, and then the monoclonal antibody was obtained with hybridoma technique. The titer of the IgG monoclonal antibody was up to 1:4 x 10(5).</p><p><b>CONCLUSION</b>Monoclonal antibody against recombinant HBoV VP2 protein was prepared and the antibody titer was high. This work may provide a new method in rapid diagnosis and study of HBoV.</p>

Effects of immediate and delayed mild hypothermia on endogenous antioxidant enzymes and energy metabolites following global cerebral ischemia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The optimal time window for the administration of hypothermia following cerebral ischemia has been studied for decades, with disparity outcomes. In this study, the efficacy of mild brain hypothermia beginning at different time intervals on brain endogenous antioxidant enzyme and energy metabolites was investigated in a model of global cerebral ischemia.</p><p><b>METHODS</b>Forty-eight male Sprague-Dawley rats were divided into a sham-operated group, a normothermia (37°C - 38°C) ischemic group and a mild hypothermic (31°C - 32°C) ischemia groups. Rats in the last group were subdivided into four groups: 240 minutes of hypothermia, 30 minutes of normothermia plus 210 minutes of hypothermia, 60 minutes of normothermia plus 180 minutes of hypothermia and 90 minutes of normothermia plus 150 minutes of hypothermia (n = 8). Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model for 20 minutes and mild hypothermia was applied after 20 minutes of ischemia. Brain tissue was collected following 20 minutes of cerebral ischemia and 240 minutes of reperfusion, and used to measure the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and adenosine triphosphate (ATP).</p><p><b>RESULTS</b>Mild hypothermia that was started within 0 to 60 minutes delayed the consumption of SOD, GSH-Px, GSH, and ATP (P < 0.05 or P < 0.01) in ischemic tissue, as compared to a normothermic ischemia group. In contrast, mild hypothermia beginning at 90 minutes had little effect on the levels of SOD, GSH-Px, GSH, and ATP (P > 0.05).</p><p><b>CONCLUSIONS</b>Postischemic mild brain hypothermia can significantly delay the consumption of endogenous antioxidant enzymes and energy metabolites, which are critical to the process of cerebral protection by mild hypothermia. These results show that mild hypothermia limits ischemic injury if started within 60 minutes, but loses its protective effects when delayed until 90 minutes following cerebral ischemia.</p>

Comparative antiplaque and antigingivitis efficacy of three antiseptic mouthrinses: a two week randomized clinical trial

The objective of this randomized, examiner blind, parallel group, controlled clinical trial was to compare the antiplaque and antigingivitis efficacy of an essential oil-containing mouthrinse (EO) to two mouthrinses containing 0.05 percent Cetylpyridinium Chloride (CPC), one with alcohol and one alcohol-free, using a two-week experimental gingivitis validated-model with a 5 percent hydroalcohol rinse serving as the negative control. One hundred and fifty-nine subjects, 56 males and 103 females; ranging in age from 18 to 58 years in good general health were assigned to one of the four treatment groups: EO (n = 40), 0.05 percent CPC with alcohol (CPCa, n = 39), 0.05 percent CPC alcohol-free (CPCna, n = 40), and 5 percent hydroalcohol negative control (n = 40). The Mean Turesky Modification of the Quigley-Hein Plaque Index (PI) and the Mean Modified Gingival Index (MGI) were the primary efficacy endpoints and were evaluated at baseline and at two weeks. Following baseline examinations, subjects received a complete dental prophylaxis and began supervised rinsing with their assigned mouthrinse twice daily for two weeks, as their sole oral hygiene measure; 151 subjects completed the trial. Two weeks after baseline the EO adjusted mean PI and MGI scores were significantly lower than those of both CPC rinses and negative control (p < 0.001). In conclusion, the EO mouthrinse demonstrated significantly greater antiplaque and antigingivitis efficacy than both CPC-containing mouthrinses and the negative control.

Studies on porcine reproductive and respiratory syndrome virus morphogenesis in Marc-145 cell and the ultrastructural changes of the infected cells / 病毒学报

Electron microscopy was employed for ultrastructural observation of Marc-145 cells infected with porcine reproductive and respiratory syndrome virus (PRRSV) SC1 strain and studied the virus morphogenesis in infected cells. The results demonstrated that PRRSV was spherical and enveloped. The virion is 45-65 nm in diameter and its nucleocapsid was approximately 25-30 nm. PRRSV entered Marc-145 cells by endocytosis, and replicated in the cytoplasm. The mature viruses were released from infected cells by budding or exocytosis. The main ultrastructural changes of the infected cells were as follows: increased number of cytoplasmic vacuoles, dilated endoplasmic reticulum, mitochondria underwent hyperplasia with its ridges swollen, sloughed, and eventually vacuolated. Typical apoptosis was also observed in the infected Marc-145 cells, which included microvilli sloughing off the cell, appearance of apoptotic bodies and cell fragmentation.

The advantages for Snail expression to promote cell migration and induce actin reorganization and to protect against the serum-deprivation-triggered apoptosis of bone marrow stem cells / 生物工程学报

The Snail transcription factor has been described as a strong repressor of E-cadherin and its stable expression induces epithelial-mesenchymal transitions responsible for the acquisition of motile and invasive properties during tumor progression. A fascinating analogy that has been raised is the seemingly similar and shared characteristics of stem cells and tumorigenic cells, which prompted us to investigate whether the mechanisms of the acquisition of invasiveness during tumor progression are also involved in bone marrow stem cells (MSCs). In this study, we examined whether Snail gene expression acts in the mobility, cytoskeleton and anti-apoptosis of MSCs. Cell Transmigration Assay and Western Blotting were performed to evaluate the cell migratory capability and the related Signaling pathways in MSCs transfected with the Snail expression vector of pCAGGSneo-SnailHA (MSCs-Sna), compared with MSCs(MSCs-neo) transducted with the control vector(pCAGGSneo). Actin cytoskeleton by Immunofluorescence and Sub-G1 detection by a FACScan flow cytometer were performed to analyze the cytoskeleton and antiapoptotic capability of MSCs-Sna. Compared with MSCs-neo, MSCs-Sna show significantly more migration in the transwell migration system (P < 0.05). And suppression of PI-3K activation by the specific PI-3K inhibitor, Wortmannin, brought on a reduction in Snail-mediated MSCs migration. In addition, we provide evidences that high expression of Snail inhibited the serum-deprivation triggered apoptosis and cytoskeleton changement of MSCs. These data suggest the possibility of facilitating MSCs migration to injured tissue and subsequent survival and maintenance in the local microenvironment after their transplantation, by investigating and increasing the advantage factors such as Snail high expression in MSCs.

Effect of low frequency stimulation and rehabilitation training on stroke patients / 中国康复理论与实践

@#ObjectiveTo investigate the effect of low frequency stimulation on motor function of stroke patients.Methods50 stroke patients were randomly divided into control group and treatment group with 25 cases in each group. Patients of control group received neuromedical and Bobath technical treatment; cases of treatment group received low frequency stimulation besides the above two methods. All patients were assessed respectively with the Fugl-Meyer Assessment and Clinical Nerve Function Impaired Scale at the admission and on 20 days after treatment.ResultsThe motor functions of both groups were improved (P<0.05). But the recovery of treatment group was superior to the control group (P<0.05).ConclusionLow frequency stimulation can promote the stroke patients' motor function recovery.

Clinical study of locally moderate hypothermia on treating acute intracerebral hemorrhage / 中国康复理论与实践

@#ObjectiveTo demonstrate the effect of moderate hypothermia and doule carotid ice applied to acute intracerebral hemorrhage (ICH) and the best opportunity of treatment.Methods174 cases with ICH were randomly divided into treated group and control group, 87 cases for each group. Each case of both groups was treated by routine therapy, moreover, all cases in treated group were treated by moderate hypothermia (MHT).The clinical effect and prognosis between treated group and control group were contrasted,so to do between the cases in the treated group who were treated no more than 3 hours and more than 3 hours after attack.ResultsCompared with the control group, the neurological function improved in the treated group(P<0.05), as well as survival rate and recovery(P<0.05).Cases who treated no more than 3 hours after attack was better than that of more than 3 hours(P<0.05).ConclusionModerate hypothermia can effectively ease the neurologic deficits after ICH, decrease mortality and disable rate.The best therapy time is within 3 hours after ICH.

Protective effect of ginkgo biloba extract on cerebral ischemia/reperfusion injury in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Ginkgo biloba extract (GbE) on dynamic equilibrium of free radicals and amino-acids in cortex of rats with cerebral ischemia/reperfusion (I/R) injury and its influence and characteristics to intracellular free calcium concentration ([Ca2+]i) in primary cultured hippocampal neuron of rats.</p><p><b>METHODS</b>Amino-acids were quantified by high performance liquid chromatographic (HPLC) analysis. Concentration of MDA and GSH-Px were determined by thiobarbituric acid (TBA) technique. SOD was assayed through xanthine method, and microfluoremetric technique was used to assay the change of [Ca2+]i and its characteristics.</p><p><b>RESULTS</b>Compared with the non-treated groups, at all time points (3 hrs after ischemia, 1 and 2 hrs after I/R separately), in the GbE treated groups, the levels of Glu, Asp and MDA were lower and SOD and GSH-Px were higher (P < 0.01 or P < 0.05), the GABA and Gly levels were lower in groups treated with middle (10 mg/kg) or high dosage (15 mg/kg) of GbE (P < 0.05). Compared with the group treated with small dosage GbE (5 mg/kg), Glu, Asp and MDA were lower and GABA, Gly, SOD and GSH-Px were higher in the groups treated with middle or high dosage of GbE (P < 0.05), while the difference in the latter two groups was insignificant. Level of [Ca2+]i in cultured neurons treated with 1 x 10(-5) mol/L glutamate combined 25 micrograms/ml GbE for 20s was lower with lower peak value and longer time for reaching the peak than that in neurons treated with 1 x 10(-5) mol/L glutamate alone. Besides, the time of decline phase was also shorter in the former, so the flatform stage was prolonged. The response was recovered by re-applying of glutamate after [Ca2+]i back to base line.</p><p><b>CONCLUSION</b>GbE can protect damaged neurons through keeping the balance of inhibitory/excitatory amino-acids, enhancing free radicals scavengers system, and inhibiting the effect of glutamate to [Ca2+]i.</p>

Development and clinical application of the EMG information management system / 中国医疗器械杂志

This paper introduces an EMG multi-gateway analysis diagnosis and information management system. The clinical applications show that this system has higher efficiency and standard report contents, and easy statistical analysis. And it also offers EMG standard figure, normal value data, nerve and muscle select scheme etc, for reference.