PDCD10-Deficiency Promotes Malignant Behaviors and Tumor Growth via Triggering EphB4 Kinase Activity in Glioblastoma.

We previously reported an angiogenic and tumor-suppressor-like function of programmed cell death 10 (PDCD10) in glioblastoma (GBM). However, the underlying mechanism remains to be elucidated. We hypothesized that loss of PDCD10 activates GBM cells and tumor progression via EphB4. To this end, PDCD10 was knocked down in U87 and T98g by lentiviral mediated shRNA transduction (shPDCD10). GBM cell phenotype in vitro and tumor growth in a mouse xenograft model were investigated in presence or absence of the treatment with a specific EphB4 kinase inhibitor NVP-BHG712 (NVP). We demonstrated that knockdown of PDCD10 in GBM cells significantly upregulated the mRNA and protein expression of EphB4 accompanied by the activation of Erk1/2. EphB4 kinase activity, reflected by phospho-EphB4, significantly increased in shPDCD10 GBM cells, and in tumors derived from shPDCD10 GBM xenografts, which was abolished by the treatment with NVP. Furthermore, NVP treatment significantly suppressed PDCD10-knockdown mediated aggressive GBM cell phenotype in vitro and extensive tumor cell proliferation, the tumor neo-angiogenesis, and a quick progression of tumor formation in vivo. In summary, loss of PDCD10 activates GBM cells and promotes tumor growth via triggering EphB4. Targeting EphB4 might be an effective strategy particularly for the personalized therapy in GBM patients with PDCD10-deficiency.

Selected Blood Inflammatory and Metabolic Parameters Predicted Successive Bilateral Sudden Sensorineural Hearing Loss.

OBJECTIVES: To explore whether peripheral inflammatory, metabolic, and hemostatic parameters could predict the pathogenesis of successive bilateral sudden sensorineural hearing loss (SSNHL). METHODS: This study reviewed 33 patients with successive bilateral SSNHL and 215 patients with unilateral SSNHL. Clinical characteristics and hematological parameters were compared, including the inflammatory markers (like neutrophil lymphocyte ratio (NLR), monocyte lymphocyte ratio (MLR), and platelet lymphocyte ratio (PLR)) and metabolic features (including hypertension, triglyceridemia, dyslipidemia, and hyperglycemia), as well as hemostatic indices (including prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen). RESULTS: In the successive bilateral SSNHL group, older average onset age (48.67 ± 15.36 vs. 42.71 ± 13.58, p < 0.05), higher male to female ratio (18 : 15 vs. 112 : 103, p > 0.05), and poorer therapeutic efficacy (12% vs. 59%, p < 0.01) were observed than those in the unilateral SSNHL group. Compared to the unilateral SSNHL group, NLR, MLR, and PLR in the successive bilateral SSNHL group were significantly higher (NLR: 5.72 ± 2.23 vs. 4.45 ± 2.82, p = 0.01; MLR: 0.25 ± 0.15 vs. 0.17 ± 0.11, p < 0.01; PLR: 190.70 ± 69.79 vs. 148.18 ± 65.67; p < 0.01); the LDL level was significantly higher; yet, the HDL level was significantly lower (LDL: 3.79 ± 0.53 vs. 3.49 ± 0.74; HDL: 1.33 ± 0.32 vs. 1.44 ± 0.26; p < 0.05 for both); fibrinogen was significantly higher (4.03 ± 0.47 vs. 3.70 ± 0.65; p < 0.01). Logistic regression analysis demonstrated that the risk factors for successive bilateral SSNHL included age, NLR, MLR, PLR, LDL, HDL, diabetes, and fibrinogen. However, only NLR, MLR, PLR, diabetes, LDL, and HDL independently predicted successive bilateral SSNHL. CONCLUSION: Selected blood inflammatory markers combined with metabolic parameters were positively correlated with successive bilateral SSNHL.

Milk Fat Globule-Epidermal Growth Factor-Factor 8 Reverses Lipopolysaccharide-Induced Microglial Oxidative Stress.

Oxidative stress plays an important role in various neurological disorders. Milk fat globule-epidermal growth factor-factor 8 (MFG-E8) is a regulatory protein for microglia. However, its involvement in microglial oxidative stress has not been established. In this study, we observed microglial oxidative stress in response to lipopolysaccharide (LPS) both in vitro and in vivo. LPS induced significant elevation of TNF-α, IL-6, MDA, and ROS and reduction of GSH and SOD in the mouse brains and primary microglia, which were reversed by MFG-E8 pretreatment. MFG-E8 induced the expression of Nrf-2 and HO-1 that was reduced by LPS incubation. Moreover, LPS-increased Keap-1 expression was reversed by MFG-E8. But the above tendencies were not seen when MFG-E8 was applied alone. The current study established the involvement of MFG-E8 in antioxidant effects during neuroinflammation. It may achieve the effects through the regulation of Keap-1/Nrf-2/HO-1 pathways.

Auditory and speech function after cochlear implantation in prelingually deaf children with white matter lesions.

AIM: To explore how auditory and speech function developed post-cochlear implantation (pCI) in prelingually deaf children with white matter lesions (WML). METHOD: Patients (41 males, 29 females; mean age at implantation 2y 11mo [SD 7.5mo], range 1y 8mo-5y) were divided into the following groups according to preoperative brain magnetic resonance imaging evaluation: mild WML, moderate WML, severe WML, and control. We assessed auditory and speech performance at baseline, 6 months, 12 months, and 24 months pCI. As well as auditory event-related potentials, topographic maps and electroencephalography source imaging were recorded and analysed at 24 months pCI. RESULTS: For children with WML (any level), postoperative auditory or speech performance at 6 months was significantly below that of control participants. After stratification, auditory and speech performance was highly related to WML grading. Auditory or speech performance in mild WML or control groups was comparatively better than the moderate WML and severe WML groups. The recovery rate of speech performance fell behind that of the auditory perception. With the increasing severity of WML, N1 amplitude was significantly smaller with a consistent presentation in the topographic map, which was similar in the mild WML and control group. The dominant auditory centre was activated in the control or mild WML groups, but not in the moderate WML and severe WML groups. INTERPRETATION: WML gradually affect auditory and speech development, and electrophysiological performance pCI in prelingually deaf children. WHAT THIS PAPER ADDS: Auditory and speech performance in prelingually deaf children with white matter lesions (WML) was significantly worse than those without WML. Postoperative auditory and speech performance in children with WML was highly related to WML grading. Grand N1 amplitude in auditory event-related potentials was negatively related to the severity of WML. Non-dominant areas close to the auditory cortex were potentially activated in severe WML.

FUNCIÓN AUDITIVA Y DEL HABLA DESPUÉS DE LA IMPLANTACIÓN COCLEAR EN NIÑOS SORDOS PRE VERBALES CON LESIONES DE MATERIA BLANCA: OBJETIVO: Explorar cómo se desarrolla la función auditiva y del habla post implante coclear (pIC) en niños sordos pre verbales con lesiones de sustancia blanca (LSB) METODO: Los pacientes (41 varones, 29 mujeres; edad media de implantación 2y 11m [DS 7.5m], rango 1y 8m - 5 años) se dividieron en los siguientes grupos de acuerdo con la evaluación de imagen de resonancia magnética cerebral preoperatoria: LSB leve, LSB moderada, LSB grave y control. Evaluamos el rendimiento auditivo y del habla al inicio del estudio, a los 6 meses, a los 12 meses y a los 24 meses, pIC. Además de los potenciales evocados auditivos, se registraron y analizaron los mapas topográficos y las imágenes de fuentes de electroencefalografía a los 24 meses pIC. RESULTADOS: Para los niños con LSB (cualquier nivel), el rendimiento auditivo o del habla postoperatorio a los 6 meses fue significativamente inferior al de los participantes de control. Después de la estratificación, el desempeño auditivo y del habla estuvo altamente relacionado con la calificación de la LSB. El rendimiento auditivo o del habla en LSB leve o en el grupo control fue comparativamente mejor que en los grupos de LSB moderada y de LSB grave. La tasa de recuperación del rendimiento del habla quedó por detrás de la percepción auditiva. Con el aumento de la gravedad de LSB, la amplitud de N1 fue significativamente menor con una presentación consistente en el mapa topográfico, que fue similar en el grupo control y de LSB leve. El centro auditivo dominante se activó en el grupo control o de LSB leve, pero no lo hizo en los grupos de LSB moderada y severa. INTERPRETACION: La LSB afecta gradualmente el desarrollo del habla auditiva y el rendimiento electrofisiológico pIC en niños sordos pre verbales.

FUNÇÃO AUDITIVA E DE FALA APÓS IMPLANTE COCLEAR EM CRIANÇAS SURDAS PRÉ-LINGUAGEM COM LESÕES DA SUBSTÂNCIA BRANCA: OBJETIVO: Explorar como a função auditiva e da fala se desenvolveu pós implante coclear (pIC) em crianças surdas pré-linguagem com lesões da substância branca (LSB). MÉTODO: Pacientes (41 do sexo masculino, 29 do sexo feminino; média de idade no implante 2a 11m [DP 7,5m], variação 1a 8m-5a) foram divididos nose seguintes grupos de acordo com a avaliação da imagem de ressonância magnética pré-operatória: LSB leve, LSB moderada, LSB severa, e controle. Avaliamos o desempenho auditivo e de fala na linha de base, 6 meses, 12 meses, e 24 meses, pCI. Também foram registrados e analisados potenciais auditivos relacionados a eventos, mapas topográficos e imagem de fontes de eletroencefalorafia 24 meses pIC. RESULTADOS: Para crianças com LSB (qualquer nível), o desempenho auditivo e de fala pós-operatório aos 6 meses foi significantemente abaixo dos participantes controle. Após estratificação, o desempenho se relaciou fortemente com o grau de LSB. O desempenho auditivo e de fala nos grupos com LSB leve e controle foi comparativamente melhor do que nos grupos LSB moderado e severo. A taxa de recuperação do desempenho da fala ficou atrás da percepção auditiva. Com a maior severidade da LSB, a amplitude N1 foi significativamente menor, com apresentação consistente no mapa topográfico, que foi similar nos grupos LSB leve e controle. O centro auditivo dominante estava ativado nos grupos controle e LSB leve, mas não nos grupos com LSB moderada e grave. INTERPRETAÇÃO: A LSB gradualmente afeta o desenvolvimento auditivo e de fala, e o desempenho eletrofisiológico pIC em crianças surdas pré-linguagem.

Loss of programmed cell death 10 activates tumor cells and leads to temozolomide-resistance in glioblastoma.

PURPOSE: Glioblastoma (GBM) is one of the most aggressive and incurable primary brain tumors. Identification of novel therapeutic targets is an urgent priority. Programmed cell death 10 (PDCD10), a ubiquitously expressed apoptotic protein, has shown a dual function in different types of cancers and in chemo-resistance. Recently, we reported that PDCD10 was downregulated in human GBM. The aim of this study was to explore the function of PDCD10 in GBM cells. METHODS: PDCD10 was knocked down in three GBM cell lines (U87, T98g and LN229) by lentiviral-mediated shRNA transduction. U87 and T98g transduced cells were used for phenotype study and LN229 and T98g cells were used for apoptosis study. The role of PDCD10 in apoptosis and chemo-resistance was investigated after treatment with staurosporine and temozolomide. A GBM xenograft mouse model was used to confirm the function of PDCD10 in vivo. A protein array was performed in PDCD10-knockdown and control GBM cells. RESULTS: Knockdown of PDCD10 in GBM cells promoted cell proliferation, adhesion, migration, invasion, and inhibited apoptosis and caspase-3 activation. PDCD10-knockdown accelerated tumor growth and increased tumor mass by 2.1-fold and led to a chemo-resistance of mice treated with temozolomide. Immunostaining revealed extensive Ki67-positive cells and less activation of caspase-3 in PDCD10-knockdown tumors. The protein array demonstrated an increased release of multiple growth factors from PDCD10-knockdown GBM cells. CONCLUSIONS: Loss of programmed cell death 10 activates tumor cells and leads to temozolomide-resistance in GBM, suggesting PDCD10 as a potential target for GBM therapy.

Do Selected Blood Inflammatory Markers Combined with Radiological Features Predict Proliferation Index in Glioma Patients?

BACKGROUND: The tumor microenvironment is partially characterized by a state of chronic inflammation, and radiologic features are related to the tumor's biological behavior. This study was conducted to explore whether peripheral blood inflammatory markers combined with radiologic features could predict proliferation potency. METHODS: This study retrospectively reviewed 183 patients with a primary diagnosis of glioma. Clinical characteristics, preoperative peripheral full blood count data, and brain magnetic resonance imaging findings were reviewed to analyze the expression of inflammatory markers neutrophil lymphocyte ratio (NLR), monocyte lymphocyte ratio, and platelet lymphocyte ratio (PLR), as well as radiologic features such as location, peritumor edema, and contrast enhancement. Immunohistochemical staining was performed to determine the proliferation index (i.e., expression of Ki-67). Receiver operating characteristic curves for cutoff value, various bivariate tests, and binary logistic regression analyses were applied. RESULTS: Proliferation index was highly associated with tumor grade, showing a gradually increasing tendency. A Ki-67 cutoff value >9% predicted high-grade glioma (HGG). Mean NLR and PLR were significantly higher in the HGG group compared with the low-grade glioma group (NLR: 3.11 ± 0.59 vs. 4.27 ± 1.13; PLR: 133.07 ± 13.17 vs. 161.51 ± 38.99; P < 0.01 for both). Contrast enhancement was more likely in the HGG group, but there was no significant between-group difference in peritumor edema. Logistic regression analysis identified the following risk factors for prediction of proliferation potency: age, Karnofsky Performance Score, NLR, PLR, and contrast enhancement. However, age >43 years, NLR >3.68, and positive contrast enhancement independently predicted a higher proliferation rate. CONCLUSIONS: NLR and contrast enhancement were positively correlated with the proliferation potency of gliomas.

The effect of allogeneic cardiac stem cells in left ventricular geometry and function in a rat model of myocardial infarction.

BACKGROUND: The development of heart failure after acute myocardial infarction (MI) was related to left ventricular (LV) pathological remodeling and dysfunction. Cardiac stem cells (CSCs) provided a new option to treat acute MI. This study was to investigate the effects of CSCs on structural and functional alteration in acute MI. METHODS: Acute MI was induced in 20 Sprague-Dawley rats by ligation of the left anterior descending coronary artery. Two weeks after MI, animals were randomized into CSCs or control group. LV geometry and function were echocardiographically measured at baseline, 2, 4, and 6 weeks after MI. After measuring hemodynamics at 6 weeks after MI, hearts were harvested for tracing CSCs stained by PKH26 and testing expression of VEGF-α/TGF-ß1 by RT-PCR and ELISA. RESULTS: Two weeks after MI, there were significant decreases in interventricular septal systolic and diastolic thickness (IVSTs/d), while increases in LV systolic and diastolic dimension (LVDs/d). Consequently, this contributed to decreases in ejection fraction (EF) and fractional shorting (FS). With the treatment of CSCs for 4 weeks, significant better ejection fraction (EF) and fractional shorting (FS) were achieved in CSCs group accompanied by the reduction in LV systole and diastole dimension (LVDs/d). Besides, a significant improvement in the maximal rate of LV pressure development and decline (peak +dP/dt and -dP/dt, respectively) was observed. Moreover, significantly higher VEGF-α was expressed in CSCs group rather than TGF-ß1. CONCLUSION: CSCs significantly prevented structural and functional deterioration after MI with increasing expression of VEGF-α.

Combination treatment with fasudil and clioquinol produces synergistic anti-tumor effects in U87 glioblastoma cells by activating apoptosis and autophagy.

Survival of patients with glioblastoma (GBM) remains poor, and novel treatment methods are urgently needed. In this study, we tested the effects of a combination of fasudil, a ROCK inhibitor, and clioquinol, an 8-hydroxyquinoline derivative with antimicrobial properties, on human GBM U87 cells. Combination treatment synergistically inhibited the viability of glioma cells but not mouse normal neuron HT22 cells and significantly induced mitochondria-mediated apoptosis. Moreover, the combination was also found to trigger macro-autophagy (henceforth referred to as autophagy) by increasing the expression levels of several proteins involved in the induction of autophagy. Further studies showed that 3-methyladenine (3-MA) or chloroquine (CQ), two autophagy inhibitors, abrogated the cytotoxic effects of the combination treatment as well as the autophagy. Overall, we demonstrated that fasudil and clioquinol show synergistic anti-cancer effects, providing evidence for the further development of combination therapy for GBM.

Combined treatment of fasudil and glutamate decreased the viability of human glioblastoma cells by excitotoxicity through NMDAR in vitro.

Glioblastoma (GBM) is the most common brain tumor with high abilities of proliferation, migration and invasion. As is well-known, the peritumoral excitotoxic neuronal cell loss caused by glutamate, secreted by GBM cells, through activated N-methyl-D aspartate receptor (NMDAR) of neuronal cell. What's more, glutamate benefits the migration of GBM cells. However, the glutamate will not kill the GBM cells itself, which may be due to the deficiency of NMDAR. Fasudil, a ROCK inhibitor, was applied for subarachnoid hemorrhage (SAH) in clinic for many years. And it was found to be of potential to inhibit the proliferation, migration and invasion of GBM cells. In present study, we applied fasudil on the primary human GBM cells to further investigate the reduction of cell viability combined with glutamate. Combination treatment of glutamate and fasudil could significantly decrease the cell viability and elevate the level of LDH compared with fasudil treatment alone. What's more, MK-801, a NMDAR antagonist, could partially abolish this death caused by combination treatment. Further study found that the expression level of NMDAR-2B was elevated after treatment with fasudil in GBM cells. These results demonstrated fasudil could increase the expression level of NMDAR, which is necessary for glutamate to work. In a word, our research has provided a new sight of medicine combination in the treatment of GBM.

CXCL12-induced upregulation of FOXM1 expression promotes human glioblastoma cell invasion.

Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor; it is highly aggressive and is associated with a poor prognosis. Binding of the chemokine CXCL12 to its receptors (CXCR4 and CXCR7) contributes to the activation of many downstream signaling pathways and promotes the invasion of various malignant tumor cells, including GBM cells. FOXM1, a transcription factor involved in cell cycle regulation, is overexpressed in GBM and is involved in GBM progression. However, the molecular mechanisms by which CXCL12 promotes the invasion of human GBM cells remain unclear. In this study, we demonstrate that CXCL12 increases the production of FOXM1 by binding to CXCR4 in GBM cell lines. Furthermore, pretreatment with an inhibitor of the PI3K/AKT pathway abrogated the CXCL12-induced expression of FOXM1. In addition, there was a positive correlation between CXCL12/CXCR4 expression and FOXM1 expression in human malignant glioma tissues. Finally, a functional assay revealed that CXCL12 does not stimulate GBM cell invasion when FOXM1 expression is silenced using a small interfering RNA (siRNA). Collectively, these findings suggest that CXCL12 promotes GBM cell invasion in part by increasing the expression of FOXM1, which is mediated in part by a PI3K/AKT-dependent mechanism in vitro.

Comparison of cardiac stem cells and mesenchymal stem cells transplantation on the cardiac electrophysiology in rats with myocardial infarction.

INTRODUCTION: Whether transplanted cardiac stem cells (CSCs) and mesenchymal stem cells (MSCs) improved ventricular fibrillation threshold (VFT) similarly is still unclear. We sought to compare the effects of the CSC and MSC transplantation on the electrophysiological characteristics and VFT in rats with myocardial infarction (MI). METHODS: MI was induced in 30 male Sprague-Dawley rats. Two weeks later, animals were randomized to receive 5 × 10(6) CSCs labeled with PKH26 in PBS or 5 × 10(6) MSCs labeled with PKH26 in phosphate buffer solution(PBS) or PBS alone injection into the infarcted anterior ventricular free wall. Six weeks after the injection, electrophysiological characteristics and VFT were measured. Labeled CSCs and MSCs were observed in 5 µm cryostat sections from each heart. RESULTS: Malignant ventricular arrhythmias were significantly (P = 0.0055) less inducible in the CSC group than the MSC group. The VFTs were improved in the CSC group compared with the MSC group. Labeled CSCs and MSCs were identified in the infarct zone and infarct marginal zone. Labeled CSCs expressed Connexin-43, von Willebrand factor, α-smooth muscle actin and α-sarcomeric actin,while the Labeled MSCs expressed von Willebrand factor, α-smooth muscle actin and α-sarcomeric actin in vivo. CONCLUSIONS: After 6 weeks of cell transplantation, CSCs are superior to MSCs in modulating the electrophysiological abnormality and improving the VFT in rats with MI. CSCs and MSCs express markers that suggest muscle, endothelium and vascular smooth muscle phenotypes in vivo, but MSCs rarely express Connexin-43.

The effects of a newly developed miniaturized mechanical chest compressor on outcomes of cardiopulmonary resuscitation in a porcine model*.

OBJECTIVE: When the duration of cardiac arrest is prolonged, reperfusion of the vital organs by effective chest compression is the most important intervention for successful resuscitation. We investigated the effects of a newly developed miniaturized chest compressor on the outcomes of cardiopulmonary resuscitation. DESIGN: Prospective, randomized, controlled experimental study. SETTING: University-affiliated animal research laboratory. SUBJECTS: Thirty male domestic pigs. INTERVENTIONS: Ventricular fibrillation was induced in 30 male domestic pigs weighing 35 ± 2 kg. Cardiopulmonary resuscitation was initiated after 7 mins of untreated ventricular fibrillation. The animals were randomized to receive mechanical chest compression with a miniaturized chest compressor, a LUCAS device or a Thumper device. After 5 mins of cardiopulmonary resuscitation, a 150-J defibrillation was delivered. If resuscitation was not successful, cardiopulmonary resuscitation was continued for 2 mins before the next defibrillation. The protocol was continued until successful resuscitation or for a total of 15 mins of cardiopulmonary resuscitation. The animals were observed for 72 hrs after resuscitation. MEASUREMENTS AND MAIN RESULTS: The miniaturized chest compressor generated significantly greater coronary perfusion pressure, end-tidal PCO2, carotid blood flow, and intrathoracic negative pressure, with significantly lower compression depth and fewer rib fractures when compared with both the LUCAS and Thumper devices. Both the miniaturized chest compressor and LUCAS devices required lower numbers of defibrillation for successful resuscitation when compared with the Thumper device. This was associated with lower prevalence of recurrent ventricular fibrillation and better postresuscitation myocardial and neurological function when compared with the Thumper device. CONCLUSIONS: The miniaturized chest compressor improves hemodynamic efficacy and the success of cardiopulmonary resuscitation with significantly less injury, which is as effective as the LUCAS device. It may provide a new option for cardiopulmonary resuscitation.

Cannabinoid 1 (CB1) receptor mediates WIN55, 212-2 induced hypothermia and improved survival in a rat post-cardiac arrest model.

AIM: The nonselective Cannabinoid (CB) receptor agonist, WIN55, 212-2, was demonstrated to induce hypothermia and improve post-resuscitation outcomes in a rat post-cardiac arrest model. The present study was to explore the potential mechanisms of WIN55, 212-2 on thermoregulation following resuscitation and to investigate which class of CB receptors was involved in WIN55, 212-2-induced hypothermia. METHODS: Ventricular fibrillation (VF) was induced and untreated for 6 min in 20 male Sprague-Dawley rats. Defibrillation was attempted after 8min of Cardiopulmonary resuscitation (CPR). Five min post-resuscitation, resuscitated animals were randomized to receive an intramuscular injection of selective CB1 receptors antagonist, SR141716A (5 mg kg(-1)); selective CB2 receptors antagonist SR144528 (5 mg kg(-1)); or placebo. Thirty min after injection, animals received continuous intravenous infusion of WIN55, 212-2 (1.0 mgkg(-1) h(-1)) for 4h while control animals received placebo. The identical temperature environment was maintained in all animals. RESULTS: In animals treated with WIN55, 212-2, blood temperatures decreased progressively from 37 °C to 34 °C within 4h. This hypothermic effect was completely blocked by CB1 but not CB2 antagonist. Accordingly, significantly better cardiac output, ejection fraction and myocardial performance index, reduced neurological deficit scores, improved microcirculation and longer duration of survival were observed in WIN55, 212-2-treated animals, which were also completely abolished by pretreatment with CB1 antagonist. CONCLUSIONS: Pharmacologically induced hypothermia with WIN55, 212-2 improved post-resuscitation myocardial and cerebral function, associated with a significantly increased duration of survival in a rat post-cardiac arrest model. The hypothermic and resulted beneficial effects of WIN55, 212-2 were mediated through CB1 receptors.

Adenovirus-mediated transfection with glucose transporter 3 suppresses PC12 cell apoptosis following ischemic injury.

In this study, we investigated the effects of adenovirus-mediated transfection of PC12 cells with glucose transporter 3 after ischemic injury. The results of flow cytometry and TUNEL showed that exogenous glucose transporter 3 significantly suppressed PC12 cell apoptosis induced by ischemic injury. The results of isotopic scintiscan and western blot assays showed that, the glucose uptake rate was significantly increased and nuclear factor kappaB expression was significantly decreased after adenovirus-mediated transfection of ischemic PC12 cells with glucose transporter 3. These results suggest that adenovirus-mediated transfection of cells with glucose transporter 3 elevates the energy metabolism of PC12 cells with ischemic injury, and inhibits cell apoptosis.

Comparison of the durations of mild therapeutic hypothermia on outcome after cardiopulmonary resuscitation in the rat.

BACKGROUND: Current studies have demonstrated that applying therapeutic hypothermia for 12 to 24 hours after resuscitation from cardiac arrest improves the outcomes of cardiopulmonary resuscitation. The present study investigated whether a shorter duration of therapeutic hypothermia induced quickly and early after resuscitation would provide an equal improvement in the outcomes of cardiopulmonary resuscitation. METHODS AND RESULTS: Ventricular fibrillation was induced and untreated for 8 minutes in 24 male Sprague-Dawley rats. Defibrillation was attempted after 8 minutes of cardiopulmonary resuscitation. Seven minutes after resuscitation, animals were randomized into 4 groups (n=6 each): normothermic, hypothermic-2 hours, hypothermic-5 hours, and hypothermic-8 hours. Animals in the hypothermic groups received rapid cooling, which was started 7 minutes after restoration of spontaneous circulation and maintained at 33±0.5°C for 2, 5, or 8 hours. Normothermic animals were maintained at 37±0.2°C. All animals were anesthetized and ventilated for 8 hours after resuscitation. Blood temperature was significantly decreased in the hypothermic groups. Postresuscitation myocardial function, neurological deficit scores, and 72-hour survival were significantly better in animals treated with hypothermia regardless of the duration of cooling. However, significantly better postresuscitation tissue microcirculation, myocardial ejection fraction, and neurological deficit scores were observed in the hypothermic-2 hours animals. CONCLUSIONS: In a rat model of cardiopulmonary resuscitation, a shorter duration of mild hypothermia induced rapidly and early after restoration of spontaneous circulation improved postresuscitation microcirculation, myocardial and cerebral functions, and survival as well as, or better than, prolonged duration of hypothermia after resuscitation.

Cholecystokinin octapeptide induces hypothermia and improves outcomes in a rat model of cardiopulmonary resuscitation.

OBJECTIVES: To investigate the effects of cholecystokinin octapeptide on thermoregulation, postresuscitation myocardial function, neurologic outcome, and duration of survival in a rat model of cardiopulmonary resuscitation. DESIGN: : Prospective, randomized, placebo-controlled experimental study. SETTING: University-affiliated animal research laboratory. SUBJECTS: Ten male Sprague-Dawley rats. INTERVENTIONS: Ventricular fibrillation was induced and untreated for 6 mins. Defibrillation was attempted after 8 mins of cardiopulmonary resuscitation. Animal temperature was adjusted to 37.0 °C with the aid of a heating lamp. At 30 mins after resuscitation, animals were randomized to receive an intravenous injection of either cholecystokinin octapeptide (200 µg/kg in 0.3 mL saline) or vehicle placebo (0.3 mL saline). The ambient temperature settings and that of the distance of the heating lamp from the animal remained the same in both groups throughout the entire experiment. MEASUREMENTS AND MAIN RESULTS: Body temperature, hemodynamic measurements, and postresuscitation myocardial function, including cardiac output, left ventricular ejection fraction, and myocardial performance index, were measured together with neurologic deficit scores and duration of survival. RESULTS: After injection of cholecystokinin octapeptide, blood temperature decreased progressively from 37.0 °C to 34.8 °C 5 hrs after resuscitation and returned to 37.0 °C at 9 hrs after injection. In the control group, blood temperature was sustained at 37.0 °C ± 0.2 °C during the same period of observation. Myocardial and neurologic function and duration of survival were significantly better in the cholecystokinin octapeptide-treated animals when compared to the control group. CONCLUSIONS: : In a rat model of cardiopulmonary resuscitation, cholecystokinin octapeptide induced mild hypothermia, attenuated postresuscitation myocardial dysfunction, and improved neurologic outcome and duration of survival.

Improvements of cardiac electrophysiologic stability and ventricular fibrillation threshold in rats with myocardial infarction treated with cardiac stem cells.

OBJECTIVES: Arrhythmia is of concern after cardiac stem cell transplantation in repairing infarcted myocardium. However, whether transplantation improved the ventricular fibrillation threshold and whether severe malignant ventricular arrhythmia is induced in the myocardial infarction model are still unclear. We sought to investigate the electrophysiologic characteristics and ventricular fibrillation threshold in rats with myocardial infarction by treatment with allogeneic cardiac stem cells. DESIGN: Prospective, randomized, controlled study. SETTING: University-affiliated hospital. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Myocardial infarction was induced in 20 male Sprague-Dawley rats. Two weeks later, animals were randomized to receive 5 × 10(6) cardiac stem cells labeled with PKH26 in phosphate buffer solution or a phosphate buffer solution-alone injection into the infarcted anterior ventricular-free wall. MEASUREMENTS AND MAIN RESULTS: Six weeks after the cardiac stem cell or phosphate buffer solution injection, electrophysiologic characteristics and ventricular fibrillation threshold were measured at the infarct area, infarct marginal zone, and noninfarct zone. Labeled cardiac stem cells were observed in 5-µm cryostat sections from each harvested heart. The unipolar electrogram activation recovery time dispersions were shorter in the cardiac stem cell group compared with those at the phosphate buffer solution group (15.5 ± 4.4 vs. 38.6 ± 14.9 msecs, p = .000177). Malignant ventricular arrhythmias were significantly (p = .00108) less inducible in the cardiac stem cell group (one of ten) than the phosphate buffer solution group (nine of ten). The ventricular fibrillation thresholds were greatly improved in the cardiac stem cell group compared with the phosphate buffer solution group. Labeled cardiac stem cells were identified in the infarct zone and infarct marginal zone and expressed Connexin-43, von Willebrand factor, α-smooth muscle actin, and α-sarcomeric actin. CONCLUSIONS: Cardiac stem cells may modulate the electrophysiologic abnormality and improve the ventricular fibrillation threshold in rats with myocardial infarction treated with allogeneic cardiac stem cells and cardiac stem cell express markers that suggest muscle, endothelium, and vascular smooth muscle phenotypes in vivo.

Optimizing the duration of CPR prior to defibrillation improves the outcome of CPR in a rat model of prolonged cardiac arrest.

AIMS: This study was to investigate whether optimal duration of CPR prior to defibrillation could be guided by Amplitude Spectrum Analysis (AMSA) in the setting of prolonged VF on outcome of CPR. METHODS: VF was induced in thirty Sprague-Dawley rats and untreated for 8 minutes. Animals were then randomized into 3 groups prior to CPR: The duration of CPR prior to defibrillation was guided by AMSA (CC+AMSA); guidelines-based with delayed defibrillation that simulated the AED algorithm (GL+AED); and guidelines-based with immediate shock (GL+shock ready). RESULTS: Regardless of groups, the majority of the animals (85%) required over 5 min of CPR to achieve restoration of spontaneous circulation (ROSC). Significantly greater rate of ROSC after first defibrillation (70% vs 0%, p < 0.01), lesser CPR interruptions and the number of defibrillations were observed in the CC+AMSA group when compared to both guidelines-based groups (p < 0.001). This was associated with a significantly better post-resuscitation myocardial and neurological function and longer durations of survival. CONCLUSIONS: After prolonged VF, optimal duration of CPR prior to defibrillation guided by AMSA improves the outcome of CPR.

Effect of therapeutic hypothermia vs δ-opioid receptor agonist on post resuscitation myocardial function in a rat model of CPR.

AIM: This study is to compare the effect of the δ-opioid receptor agonist, D-Ala(2)-D-Leu(5) enkephalin (DADLE) with normothermic control and therapeutic hypothermia on post resuscitation myocardial function and 72-h survival in a rat model of cardiac arrest and resuscitation. METHODS: Ventricular fibrillation (VF) was induced in 15 male Sprague-Dawley rats. After 8 min of untreated VF, cardiopulmonary resuscitation was performed for 8 min before defibrillation. Animals were randomized to three groups of five: (a) normothermia; (b) hypothermia (32 °C); and (c) normothermia with DADLE intravenous infusion (1 mg/kg h(-1)). Hypothermia and drug infusion were started after successful defibrillation. Myocardial functions, including cardiac output (CO), left ventricular ejection fraction (LVEF), and myocardial performance index (MPI) were measured echocardiographically together with duration of survival. RESULTS: The 72-h survival was significantly greater in the hypothermic group than in both DADLE and normothermic group (p = 0.02). However, the survival time of the DADLE treated animals was significantly longer than that of the normothermia group (51.8 ± 18.9 vs 18.8 ± 10.1h, p < 0.01). DADLE group showed significantly better CO (PR 60 min, p = 0.049), better LVEF (PR 60 min, p = 0.044; PR 240 min, p < 0.001) and lower MPI (PR 60 min, p = 0.043; PR 240 min, p = 0.045) than normothermic group. Hypothermia group also showed significantly better CO (PR 60m in, p = 0.044; PR 240 min, p = 0.007), better LVEF (PR 60 min, p = 0.001; PR 240 min, p < 0.001) and lower MPI (PR 60 min, p = 0.003; PR 240 min, p = 0.012) than the normothermic group. CONCLUSIONS: DADLE attenuated post resuscitation myocardial dysfunction and increased short term survival time. However, the 72-h survival in the DADLE group was less than that in the hypothermia group.