Serum Creatine Kinase Isoenzyme MB Research Articles

Feasibility of aligning creatine kinase MB activity and mass data in multicenter trials using generalized additive modeling.

Elevated serum creatine kinase isoenzyme MB (CK-MB) levels indicate myocardial ischaemia and periprocedural myocardial injury during treatment of heart diseases. We established a method to predict CK-MB mass from activity data based on a prospective pilot study in order to simplify multicentre trials. 38 elective cardiac surgery patients without acute myocardial ischaemia and terminal renal failure were recruited. CK-MB mass and activity were determined in venous blood samples drawn preoperatively, postoperatively, 6 h post-op, and 12 h post-op. Linear regression and generalized additive models (GAMs) were applied to describe the relationship of mass and activity. Influences of demographic and perioperative factors on the fit of GAMs was evaluated. The agreement of predicted and measured CK-MB masses was assessed by Bland-Altman analyses. Linear regression provided an acceptable overall fit (r2=0.834) but showed deviances at low CK-MB levels. GAMs did not benefit from the inclusion of age, body mass index, and surgical times. The minimal adequate model predicted CK-MB masses from activities, sex, and sampling time with an r2 of 0.981. Bland-Altman analyses confirmed narrow limits of agreement (spread: 8.87 µg/L) and the absence of fixed (p = 0.41) and proportional (p = 0.21) biases. GAM-based modelling of CK-MB data in a representative patient cohort allowed to predict CK-MB masses from activities, sex, and sampling time. This approach simplifies the integration of study centers with incompatible CK-MB data into multicentre trials in order to facilitate inclusion of CK-MB levels in statistical models.

B-Type Natriuretic Peptide Can Distinguish Ischemic Stroke Subtypes Better than Other Cardiac Biomarkers.

BACKGROUND The primary benefit to patients of being able to distinguish among subtypes of ischemic stroke (IS) is creation of a better treatment decision-making process. Current classification methods are complex and time-consuming, requiring hours to days. Blood-based cardiac biomarker measurements have the potential to improve mechanism classification of ischemic stroke. MATERIAL AND METHODS In this study, 223 patients with IS were selected as the case group and 75 healthy people who underwent physical examination at the same time were selected as the control group. The chemiluminescent immunoassay (CLIA) method established in this study was used to quantitatively detect plasma B-type natriuretic peptide (BNP) levels in subjects. All subjects were assessed for serum creatine kinase isoenzyme-MB (CK-MB), cardiac troponin I (cTnI), and myoglobin (MYO) after admission. We investigated the effectiveness of BNP and other cardiac biomarkers in the diagnosis of different subtypes of IS. RESULTS The levels of the 4 cardiac biomarkers were increased in IS patients. BNP could better diagnose different types of IS compared to other cardiac biomarkers, and BNP combined with other cardiac biomarkers was better than a single indicator in diagnosing IS. CONCLUSIONS Compared with other cardiac biomarkers, BNP is a better marker for the diagnosis of different subtypes of ischemic stroke. Routine screening for BNP in IS patients is recommended to improve the treatment decision-making process and minimize the time to thrombosis, thereby providing a more precise treatment for patients with different subtypes of stroke.

Dexmedetomidine Pretreatment Protects Against Myocardial Ischemia/Reperfusion Injury by Activating STAT3 Signaling.

Myocardial infarction is a common perioperative complication, and blood flow restoration causes ischemia/reperfusion injury (IRI). Dexmedetomidine (DEX) pretreatment can protect against cardiac IRI, but the mechanism is still insufficiently understood. In vivo, myocardial ischemia/reperfusion (30 minutes/120 minutes) was induced via ligation and then reperfusion of the left anterior descending coronary artery (LAD) in mice. Intravenous infusion of 10 μg/kg DEX was performed 20 minutes before ligation. Moreover, the α2-adrenoreceptor antagonist Yohimbine and STAT3 inhibitor Stattic were applied 30 minutes ahead of DEX infusion. In vitro, hypoxia/reoxygenation (H/R) with DEX pretreatment for 1 hour was performed in isolated neonatal rat cardiomyocytes. In addition, Stattic was applied before DEX pretreatment. In the mouse cardiac ischemia/reperfusion model, DEX pretreatment lowered the serum creatine kinase-MB isoenzyme (CK-MB) levels (2.47 ± 0.165 vs 1.55 ± 0.183; P < .0001), downregulated the inflammatory response ( P ≤ .0303), decreased 4-hydroxynonenal (4-HNE) production and cell apoptosis ( P = .0074), and promoted the phosphorylation of STAT3 (4.94 ± 0.690 vs 6.68 ± 0.710, P = .0001), which could be blunted by Yohimbine and Stattic. The bioinformatic analysis of differentially expressed mRNAs further confirmed that STAT3 signaling might be involved in the cardioprotection of DEX. Upon H/R treatment in isolated neonatal rat cardiomyocytes, 5 μM DEX pretreatment improved cell viability ( P = .0005), inhibited reactive oxygen species (ROS) production and calcium overload (both P ≤ .0040), decreased cell apoptosis ( P = .0470), and promoted STAT3 phosphorylation at Tyr705 (0.102 ± 0.0224 vs 0.297 ± 0.0937; P < .0001) and Ser727 (0.586 ± 0.177 vs 0.886 ± 0.0546; P = .0157), which could be abolished by Stattic. DEX pretreatment protects against myocardial IRI, presumably by promoting STAT3 phosphorylation via the α2-adrenoreceptor in vivo and in vitro.

Curdione Relieved Isoproterenol-Induced Myocardial Damage through Inhibiting Oxidative Stress and Apoptosis.

Isoproterenol (ISO) is widely used to treat bronchial asthma, cardiogenic or septic shock, complete atrioventricular block, and cardiac arrest. However, it can also cause myocardial damage owing to infarct-like necrosis. Curdione, an extract of the Chinese herb Rhizoma Curcumae, has a variety of pharmacological activities, including cardioprotective effects. In this study, we investigated the protective effects of curdione and its underlying mechanisms in an ISO-induced myocardial injury model. Our results showed that curdione attenuated ISO-induced H9c2 cell proliferation inhibition and lactic dehydrogenase (LDH) release. Curdione ameliorated morphological damage and reduced the ISO-induced elevation of serum creatine kinase-MB isoenzyme (CK-MB) and LDH.Furthermore, curdione inhibited ISO-induced cell apoptosis, modulated the expression of Bcl-2 and Bax proteins, repealed the accumulation of ISO-induced reactive oxygen species (ROS), prevented mitochondrial dysfunction, and activated the Nrf2/SOD1/HO-1 signaling pathway. The above results show that curdione exerts a protective effect against ISO-induced myocardial damage by inhibiting apoptosis and oxidative stress, suggesting that curdione is a potential therapeutic strategy to prevent ISO-induced myocardial damage.

Case report: One child with an autism spectrum disorder who had chronically elevated serum levels of CK and CK-MB

Some patients with autism spectrum disorder (ASD) exhibit elevated serum creatine kinase levels, which are believed to be associated with mitochondrial dysfunction. Although a few articles have reported this situation in the past and the increase mostly ranges from 100 to 300 U/L, there is a paucity of previous study focusing on the serum creatine kinase MB isoenzyme. This article discusses a 5-year-old girl with ASD, whose serum creatine kinase and creatine kinase MB isoenzyme have been rising for nearly 2 years, fluctuating at 584–993 and 111–625 U/L respectively. Except for behavioral and language symptoms associated with ASD, the child appears normal in other aspects. The child's laboratory tests showed no abnormality, except that the serum levels of lactic acid was slightly higher than normal (1.89 mmol/L, normal 1.33–1.78 mmol/L). The child was prescribed with a traditional Chinese medicine during the process and the serum creatine kinase MB isoenzyme level decreased dramatically to 111 U/L after the treatment. This study firstly recorded the serum creatine kinase levels and the MB isoenzyme in patients with autism spectrum disorder for nearly 2 years, indicating that patients with ASD may experience long-term increases in serum creatine kinase and creatine kinase MB isoenzyme, and that the traditional Chinese medicine decoction Xinfukang can temporarily reduce the serum creatine kinase MB isoenzyme level in patients. Nevertheless, the effect is not sustained. Therefore, it is of great importance to conduct long-term longitudinal studies so as to elucidate the potential mechanism responsible for long-term elevation of serum creatine kinase level.

Clinical characteristics of 68 children with atypical hand, foot, and mouth disease caused by coxsackievirus A6: a single-center retrospective analysis.

BackgroundHand, foot, and mouth disease (HFMD) caused by coxsackievirus A6 (CV-A6) has become prevalent in many parts of the world. It is commonly referred to as atypical HFMD which more likely to present as bullous lesions. Compared with traditional HFMD, its misdiagnosis rate is relatively high, which brings difficulties to clinical diagnosis. We retrospectively analyze the clinical characteristics of children with HFMD with bullous lesions caused by CV-A6.MethodsThe study included 68 children with atypical HFMD caused by CV-A6 who were hospitalized from 2018 to 2020. Data of the children including age, sex, month of HFMD onset, the morphologies and distribution of rashes, the details of fever, the presence or absence of onychomadesis, and laboratory test results were analyzed and compared between an infant group (<1 year), a toddler group (1–<3 years), and a preschool group (3–<6 years).ResultsOf the 68 children, 67 were younger than 5 years old, with a male to female ratio of 1.62:1. The disease peaked in the period from June to September. With 75.0% of the infant group had more than three kinds of rashes; 95.0% of the preschool group had rashes in more than five locations. These differences were statistically significant (P<0.05). All children had fever. The peak fever in the toddler group was lower (P=0.033). No critical cases were observed in any of the groups. Of the 61 children who were successfully followed up, 68.9% developed onychomadesis within 2–3 weeks. The proportion of cases with abnormal liver function was 83.3%, 41.7%, and 10.0% in the infant, toddler, and preschool groups (P<0.001). The proportion of cases with increased serum creatine kinase MB isoenzyme (CK-MB) were significantly higher in the toddler group (P<0.05).ConclusionsAtypical HFMD caused by CV-A6 infection usually occurred in children under 5 years old. The morphologies of the rashes in the infant group changed more, while the rashes in the preschool group was more widely distributed. The incidence of critical cases was low. More than half of the cases can develop onychomadesis in the recovery period. Organ damage was relatively mild in the preschool group.

Serum TP53 Protein Level as a Sensitive Biomarker for the Diagnosis of Myocardial Damage in Children.

BackgroundHigh levels of TP53 protein can lead to apoptosis of myocardial cells. However, TP53 protein influence of myocardial damage remains unclear. This prospective study investigated the involvement of TP53 protein in secondary myocardial damage in children up to 18 years of age.Material/MethodsSerum TP53 protein, N-terminal prohormone B-type natriuretic peptide (NT-ProBNP), cardiac troponin-I (cTnI), and creatine kinase isoenzyme MB (CK-MB) concentrations were measured in 50 hospitalized patients with secondary myocardial damage, 50 hospitalized patients without myocardial damage, and 50 healthy individuals (control). Cardiac damage was diagnosed based on cTnI, NT-ProBNP, and CK-MB levels, with electrocardiographic evidence as the reference. The appropriate cut-off value of TP53 protein for secondary myocardial damage was analyzed by receiver operating characteristic (ROC) curves.ResultsThe serum TP53 protein, NT-ProBNP, cTnI, and CK-MB concentrations of the patients with and without myocardial damage were 10.20±1.20 and 0.30±0.10 ng/L, 505.30 and 107.8 ng/L, 0.23±0.13 and 0.02±0.01 μg/L, and 28.30±5.13 and 12.24±4.29 IU/L, respectively. For the 50 patients with myocardial damage, the area under the ROC curve for serum TP53 protein, NT-ProBNP, cTnI, and CK-MB concentrations were 0.89 (95% CI: 0.81–0.95), 0.83 (95% CI: 0.77–0.91), 0.92 (95% CI: 0.84–0.97), and 0.85 (95% CI: 0.78–0.93), respectively, and the diagnostic cut-off values were 12.00 ng/L, 500.00 ng/L, 0.16 μg/L, and 27.00 IU/L, respectively, with positive likelihood ratios of 20.8, 13.2, 24.6, and 15.6.ConclusionsTP53 protein is a valid biomarker of secondary myocardial damage in pediatric patients and can be diagnostic.

Hypercholesterolemia attenuates cardioprotection of ischemic preconditioning and postconditioning with α7 nicotinic acetylcholine receptor agonist by enhancing inflammation and inhibiting the PI3K/Akt/eNOS pathway.

The present study aimed to evaluate the effects of hypercholesterolemia on cardioprotection of ischemic preconditioning and α7 nicotinic acetylcholine receptor (α7nAChR) agonist postconditioning and explore the potential mechanisms that hypercholesterolemia affected their cardioprotection. Hypercholesterolemic and normal rats were divided into the four groups that received the following treatments: i) Hypercholesterolemic control and normal control groups; ii) hypercholesterolemic ischemia/reperfusion (HI) and normal ischemia/reperfusion (NI) groups; iii) hypercholesterolemic ischemic preconditioning (HIPC) and normal ischemic preconditioning (NIPC) groups; and iv) hypercholesterolemic PNU282987 postconditioning (HPNU) and normal PNU282987 postconditioning (NPNU) groups. Serum lactate dehydrogenase (LDH), creatine kinase isoenzyme MB (CK-MB), cardiac troponin I (cTnI), tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) levels after ischemia/reperfusion were assayed. Furthermore, infarct size and expression levels of Akt, phosphorylated (p)-Akt and endothelial nitric oxide synthase (eNOS) in ischemic myocardium were assessed. Compared with the NI group, serum LDH, CK-MB, cTnI, TNF-α and IL-6 levels and infarct size were significantly decreased, and myocardial p-Akt/Akt and eNOS/GAPDH ratios were significantly increased in the NIPC and NPNU groups. Compared with the HI group, serum CK-MB, cTnI, TNF-α and IL-6 levels and infarct size were significantly decreased in the HIPC group; however, myocardial p-Akt/Akt and eNOS/GAPDH ratios did not significantly change in the HIPC group. Furthermore, there were no significant difference between the HI and HPNU groups in serum LDH, CK-MB, cTnI, TNF-α and IL-6 levels, infarct size, myocardial p-Akt/Akt and eNOS/GAPDH ratios. In conclusion, hypercholesterolemia could aggravate myocardial ischemia/reperfusion injury, attenuate cardioprotection of ischemic preconditioning and eliminate cardioprotection from α7nAChR agonist postconditioning by enhancing inflammation and inhibiting PI3K/Akt/eNOS pathway.

Preparation of Puerarin Nanocrystals and Their Application on Rats with Myocardial Ischemic Injury Combined with Electroacupuncture-PC6 (Neiguan)

The effect of electroacupuncture PC6 (Neiguan) combined with puerarin nanocrystal preparation (PUE-NP) on myocardial injury and autophagy in a rat model with myocardial ischemia was explored in this study. PUE-NPs were first prepared, and there was an analysis on the fourier transform infrared spectroscopy (FTIR), particle size distribution of nanocrystalline preparation, Zeta potential, and X-ray diffractometry (XRD) characterization, so as to analyze the difference in the in vitro cumulative release rate of PUE and PUE-NPs. Secondly, Sprague-Dawley (SD) rats were taken as the subjects investigated. Besides, they were divided randomly into a sham surgery group (Sham), a myocardial ischemia model group (Model), an electroacupuncture Neiguan acupoint pretreatment model group (EA+M), an electroacupuncture Neiguan acupoint pretreatment combined with PUE treatment model group (EA+PUE+M), and an acupuncture Neiguan point pretreatment combined with PUENPs treatment model group (EA+PUE-NPs+M). Then, the rats were treated with abdominal anesthesia, and their serums were extracted to detect the differences in serum creatine kinase MB isoenzyme (CK-MB) and lactate dehydrogenase (LDH) concentrations. After the rats were sacrificed, myocardial tissues were taken out, and the Western blot, hematoxylin-eosin (HE) staining, and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining were employed to detect apoptosis index, pathological changes of myocardial tissue morphology, and expression differences of autophagy-related protein, such as Beclin-1, LC3, and mammalian target of rapamycin (mTOR). It was found that the average particle size of PUE-NPs was 155.9±17.3 nm, and the average Zeta potential was −15.3±2.1 mV. FTIR and XRD maps displayed that PUE-NPs nanoparticles were successfully prepared, and had a longer lasting in vitro release rate compared with PUE bulk drugs. The results of animal experiments indicated that the myocardial infarction area, serum LDH and CK-MB concentration, and myocardial cell apoptosis index, LC3II/I ratio, mTOR phosphorylation level, and Beclin-1 protein expression level of rats from the EA+M group, EA+PUE+M group, and EA+PUENPs+ M group reduced sharply compared with the model group (P &lt; 0.05). Compared with the EA+M group, the above indicators of rats from the EA+PUE+M group and EA+PUE-NPs +M group dropped steeply (P &lt; 0.05). Besides, the above indicators of rats from the EA +PUE-NPs+M group were lower obviously than those indicators of the EA+PUE+M group (P &lt; 0.05). In conclusion, the above results revealed that the combination of electroacupuncture PC6 (Neiguan) and PUE preparation could improve myocardial cell apoptosis and autophagy levels in rats with myocardial ischemia, while the improvement effect of combining PUE-NPs was more marked.

Selenium mitigated aflatoxin B1-induced cardiotoxicity with potential regulation of 4 selenoproteins and ferroptosis signaling in chicks

The aim of the present study was to explore the underlying mechanism of selenium (Se)-mediated detoxification of aflatoxin B1 (AFB1)-induced cardiotoxicity in chicks. A Se-deficient, corn-soybean meal-basal diet (36 μg Se/kg, BD) and three test diets (BD+1.0 mg AFB1/kg, 0.3 mg Se/kg, or 1.0 mg AFB1/kg+0.3 mg Se/kg) were used in a 3-wk 2 × 2 factorial design trial (n = 30 chicks/group). Dietary AFB1 led to induced (P < 0.05) serum creatine kinase and creatine kinase MB isoenzyme activities and heart histopathologic lesions. However, Se deficiency aggravated most of these alterations induced by AFB1. Moreover, mRNA levels of two ferroptosis activators (solute carrier family 11 Member 2 and transferrin) were upregulated (P < 0.05) in the AFB1-treated groups. Additionally, Se deficiency reduced (P < 0.05) glutathione peroxidase (GPX) 3 and thioredoxin reductase 3 mRNA and GPX activity but increased (P < 0.05) selenoprotein M and selenophosphate synthetase 2 mRNA in the heart in AFB1-administered groups. The in vitro study showed that Se alleviated (P < 0.05) AFB1-reduced cell viability and induced (P < 0.05) ROS and ferroptosis in H9C2 cardiac cells. It also downregulated (P < 0.05) two ferroptosis activators (long-chain acyl-CoA synthetase 4 and solute carrier family 11 Member 2) in the AFB1-treated groups in the H9C2 cells. In conclusion, this study illustrated that Se alleviates AFB1-induced cardiotoxicity and cardiomyocyte damage potentially related to the regulation of redox status, 4 selenoproteins, and ferroptosis-related signaling.

Ethanolic and Aqueous Extracts of Avocado (Persea americana) Seeds Attenuates Doxorubicin-Induced Cardiotoxicity in Male Albino Rats

Cardiotoxicity from long-term use of the anticancer drug, doxorubicin (DOX), imposes a strict limitation its administration. The health benefits of avocado seeds (AS) stem from essential nutrients and phytochemicals. This study aimed to compare effects of ethanol (EAS), and water (WAS) extracts from AS on DOX-induced cardiotoxicity. Forty adult male albino rats were divided into four groups: GI were control rats. GII rats received DOX alone over a 2 week period. GIII animals received the DOX treatment plus AS. GIV rats received the DOX treatment plus EAS. AS extract treatments were administered after initiation of cardiotoxicity. Total flavonoids, polyphenols, and antioxidant capacity of both AS extracts were measured. Serum markers of cardiac dysfunction were evaluated, oxidative stress and inflammatory biomarkers were measured in cardiac tissue homogenates and histopathology of heart tissues was evaluated. The AS ethanolic extract had a higher content of active compounds. DOX treatment increased markers of cardiac dysfunction Matrix Metalloproteinase-1 (MMP-1), Serum Cardiac Troponin I (cTnI), Lactate Dehydrogenase (LDH), Serum Creatine Kinase Isoenzyme-MB (CK-MB), and Heart-Type Fatty Acid-Binding Protein (FABP3) and produced increases in levels of Advanced Oxidation Protein Products (AOPP) and Malondialdehyde (MDA) in cardiac tissue homogenates with a significant reduction in Catalase (CAT), Total Antioxidant Capacity (TAC). It also increased levels of Tumor Necrosis Factor (TNF-a), Interleukin-6 (IL-6), Interleukin-1 beta (IL-1s) and Caspase-3, in the cardiac tissue homogenates. EAS was more effective than WAS, but both acted as cardioprotective agents following DOX treatment as evidenced by mitigation of impacts on biochemical markers.

Curdione Ameliorated Doxorubicin-Induced Cardiotoxicity Through Suppressing Oxidative Stress and Activating Nrf2/HO-1 Pathway.

Doxorubicin (DOX) is a representative antibiotic of terpenoids and clinically used in the treatment of various malignant tumors. However, its application is limited by the cardiotoxocity. Curdione, an extract from Rhizoma Curcumae, has many promising pharmacological effects including protecting acute liver injury and cerebral ischemia. It is still unknown whether curdione has a protective function for DOX-induced cardiotoxicity. In our study, we investigated the protective effects of curdione against DOX-induced cardiotoxicity. Our results showed that curdione attenuated DOX-induced growth inhibition and release of lactic dehydrogenase in a concentration-dependent manner. And curdione ameliorated the histopathological damage, reduced the elevation of serum creatine kinase-MB isoenzyme (CK-MB) and lactic dehydrogenase by DOX. Furthermore, curdione inhibited DOX-induced cell apoptosis and modulated the expression of Bcl-2 and Bax proteins, as well as abrogated DOX-induced reactive oxygen species accumulation and prevented mitochondria dysfunction. Further study indicated that curdione decreased DOX-induced phosphorylation of extracellular signal-regulated kinase1/2 (Erk1/2) and c-Jun-N-terminal kinase and activated nuclear factor-erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) signal pathway. Our results suggested that curdione maybe is a new and feasible strategy to prevent DOX-induced cardiotoxicity through monitoring multiple targets.

Protective effect of supplementation with Lycium ruthenicum Murray extract from exhaustive exercise-induced cardiac injury in rats.

To the Editor: Lycium ruthenicum (L. ruthenicum) Murray contains abundant anthocyanins. The purpose of the present research was to investigate the protective effect of L. ruthenicum extract supplementation on the damage from exhaustive exercise in rats which were treated for seven days before the exhaustive exercise. Exhaustive exercise induced an increase of Serum nitrate (NO) and reactive oxygen species (ROS) levels, but supplementation of L. ruthenicum extract reduced them significantly (P < 0.05). Interleukin (IL)-1 and IL-6 were also significantly reduced by L. ruthenicum as compared to the control (P < 0.05). Supplementation with L. ruthenicum extract also decreased the creatine kinase isoenzyme MB (CK-MB) generation significantly and ameliorated the myocardial histopathology. L. ruthenicum belongs to the genus Lycium of the family Solanaceae and is a salinized desert plant distributed mainly in Qinghai and Xinjiang Provinces of China. Recently, studies have shown that L. ruthenicum exhibits a wide range of pharmacologic properties, such as anti-oxidation, anti-fatigue, and hypoglycemic activity.[1,2] Anthocyanins and their aglycones from L. ruthenicum have strong antioxidant activity.[3] Anthocyans are flavonols, which occur ubiquitously in the plant kingdom and confer bright red or blue coloration on berries and other fruits and vegetables. Anthocyanin concentration in the L. ruthenicum extract was determined by using the Nakata method[4] and the concentration in the L. ruthenicum extract was 24.1 ± 7.7 mg/g. However, we know anthocyanin has antioxidant properties and ROS induced by extreme temperatures can be protected by anthocyanins in several plants. Anthocyanin has an antioxidant role in plants against ROS and extreme temperatures. Exercise-induced oxidative stress has been reported and antioxidant supplementation can prevent muscle damage, protective effects on the endocrine-metabolic and immune profile.[5] Therefore, L. ruthenicum extract should have a protective effect against oxidative stress and we investigated that the possible protective effect of L. ruthenicum extract and anthocyanin supplementation on the oxidative stress generated by exhaustive exercise in SD rats. In this study, 30 Sprague-Dawley (SD) rats, males and aged 6 weeks, were randomly assigned to the control, anthocyanin (5 mg/rat as the positive control), and other three groups were given L. ruthenicum extract in distilled water orally (as vehicle, 10 and 30 mg/rat), for 7 days before the exhaustive exercise stress. Rats of the exhaustive exercise group were subjected to graded treadmill running started at 10% grade, 15 m/min for 10 min followed by gradual increases of treadmill speed and time as 25 m/min for 20 min, 28 m/min for 20 min, 31 m/min for 20 min, 34 m/min for 20 min, 37 m/min for 20 min until exhaustion. The heart was dissected and washed with saline for fixation with formalin. Exhaustive exercise causes oxidative stress and is defined as the inability of rats to run on the treadmill despite a mild electrical shocking.[6] Add some information on dietary antioxidants for exercise-induced oxidative stress.[7] Regular exercise is recommended for a healthy life, but exhaustive exercise generates the excessive oxidative stress that may exceed the beneficial effect from the regular exercise.[8] In the present study, an increasing CK-MB activity from the cardiac muscle consisted of myocardial damage was seen in tissue vacuoles and cell death from heart tissue. Exhaustive exercise also induced an increase of serum NO, ROS, IL-1, IL-6, and CK-MB levels, but supplementation of L. ruthenicum extract reduced them significantly. Supplementation with L. ruthenicum extract also significantly ameliorated the myocardial histopathology. Under the microscope, the photographs showed the heart section with a 400× magnification. The severity of damage in the exhaustive exercise group was exhibited by vacuoles and dead cells. However, changes in histopathology were significantly reduced by the treatment with anthocyanin and L. ruthenicum extract [Figure 1].Figure 1: Protective effect of Lycium ruthenicum extract on the exhaustive exercise-induced myocardial injury. Histopathology (hematoxylin and eosin staining) of myocardial slices from rats from (A) no treatment control group; (B) the exhaustion group; (C) the Anthocyanin group, exhaustive exercise treatment supplemented with 5 mg Anthocyanin/rat; and (D) the L. ruthenicum extract group, exhaustive exercise treatment supplemented with 30 mg L. ruthenicum/rat. Photographs show the myocardial section with 400× magnification. Tissue vacuoles (hollow triangle arrow heads ▿) and cell death (nuclei stained, black arrow heads ◂) were more severe in exhaustion than those in control or L. ruthenicum groups.In conclusion, 1-week oral supplementation of L. ruthenicum extracts present positive effects on the oxidative stress generated by exhaustive exercise in SD rats, possibly via promotion of antioxidant capacity. The mechanism of protection of L. ruthenicum extract supplementation was not investigated in this study and waits for further study in the future. Acknowledgements The authors are grateful for text proofreading support from Prof. Kee-Ching Jen. Funding None. Conflicts of interest None.

Puerarin pre-conditioning on the expression levels of CK-MB, cTnI and inflammatory factors in patients undergoing cardiac valve replacement.

Effect of puerarin preconditioning on the expression levels of nuclear factor κB (NF-κB), interleukin 6 (IL-6), interleukin 8 (IL-8), troponin I (cTnI), and creatine kinase isoenzyme MB (CK-MB) in the neutrophils of patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB) was evaluated. We enrolled 50 patients scheduled for cardiac valve replacement and assigned them randomly divided into either a puerarin or a control group. Puerarin was dissolved in 10 ml normal saline before CPB, and administered by intravenous infusion to patients in the puerarin group. The control group was administered an equivalent amount of saline. We used flow cytometry to determine the expression levels of NF-κB, IL-6 and IL-8 in neutrophils and an auto chemistry analyzer to determine the serum levels of cTnI and CK-MB before anesthesia induction (T0), 30 min after aortic declamping (T1), 4 h after aortic declamping (T2), and 8 h after aortic declamping (T3). We found the mean serum cTnI and CK-MB levels of the puerarin group tended to decrease with time. The positive rates of NF-κB, IL-6 and IL-8 at different time-points were lower in patients of the puerarin group than in those of the control group (and the differences at T3 were statistically significant). The clinical manifestations of patients in the puerarin group after operation were better than those in the control group (P<0.05). We found that the expression levels of NF-κB, IL-6 and IL-8 were positively correlated with the levels of CK-MB and cTnI (P<0.05). Puerarin preconditioning can reduce the NF-κB activation and the overexpression of IL-6 and IL-8 in neutrophils, and it inhibits the release of myocardial enzyme cTnI and CK-MB reflecting myocardial cell protection. Puerarin seems to improve safety and efficacy of valvular replacement operations.

THE EFFECTS OF CARVEDILOL AND Q10 ON ISOPRENALINE-INDUCED CARDIOTOXICITY: ELECTROCARDIOGRAPHIC, BIOCHEMICAL AND HISTOPATHOLOGICAL EVALUATION IN RATS.

Objective: To assess the therapeutic effect of combining of Carvedilol and Q10 on electrocardiographic, biochemical and histopathological changes in isoprenaline induced myocardial infarction. Methods: Myocardial infarction was induced in rats by subcutaneous (s.c) injection of isoprenaline (85mg/kg) for two consecutive days at an interval of 24 h. Rats were treated orally (p.o) with Carvedilol at dose (5 mg/kg/day) and Q10 at dose (10 mg/kg/day) for a period of 14 days. At the end of experiment electrocardiographic, biochemical and histopathological changes were monitored from control and experimental groups. Results: ISP intoxicated rats showed a significant tachycardia , elevation ST and prolongation of QTc (i.e. ST, QTc & HR) due to conduction abnormalities, increased serum creatine kinase isoenzyme-MB (CK-MB) Treatment with Carv+Q10 significantly improved the ISP induced alteration in ECG, biochemical and histopathological changes. Conclusions: The present result shows that treatment with Carv+Q10 significantly attenuates myocardial infarction in ISP-intoxicated rats.

Effects of ketamine constant rate infusions on cardiac biomarkers and cardiac function in dogs

ObjectiveTo evaluate the effects of ketamine continuous rate infusions (CRI) at two dose rates on cardiovascular function and serum creatine kinase MB isoenzyme (CK-MB) and troponin I in healthy conscious dogs. Study designExperimental, prospective, crossover, randomized, blinded study. AnimalsEight adult mixed-breed dogs, aged 6±1 years and weighing 19±8.6 kg (mean±standard deviation). MethodsDogs were administered an intravenous bolus of ketamine (0.5 mg kg−1) followed by a ketamine CRI for 12 hours (20 μg kg−1 minute−1; treatment TC20 or 40 μg kg−1 minute−1; treatment TC40). Sedation, heart rate (HR), mean arterial pressure (MAP), electrocardiographic and echocardiographic parameters were evaluated at baseline (T0) and 1 (T1), 2 (T2), 4 (T4), 8 (T8), 12 (T12) and 24 (T24) hours after ketamine infusion started. Serum concentrations of CK-MB and troponin I were measured at baseline and 12, 24 and 48 hours after infusion started. ResultsHR increased over the first 4 hours, significantly at T1 in TC20 and at T4 in TC40 when compared with T0 (p < 0.05). MAP was significantly increased at T2 in TC40 when compared with TC20. Behavioral changes, such as stereotypical head movements and twitches, occurred within 4 hours in TC40. There were no significant changes in echocardiographic examinations in any dog when compared with baseline. There were no temporal changes in serum CK-MB activity either within or between treatments (p > 0.05). No troponin I was detected in any sample. Conclusions and clinical relevanceNo indication of myocardial injury resulting from ketamine infusion was detected in this study in healthy dogs. Further studies are needed to assess the ketamine infusion effects on antinociception and other organ function not evaluated in the present study.

MiRNA-21 Expression in the Serum of Elderly Patients with Acute Myocardial Infarction.

BackgroundThe aims of this study were to examine the expression of miRNA-21 in the serum of elderly patients (>65 years) with acute myocardial infarction (AMI) and to investigate the potential role of serum miRNA-21 as a marker of early cardiac myocyte damage.Material/MethodsThirty-eight elderly patients with recent AMI, 27 elderly patients with unstable angina pectoris, and 25 healthy elderly individuals were included in the study. Serum miRNA-21 expression was determined following total RNA extraction and reverse-transcribed into cDNA, followed by reverse transcription-polymerase chain reaction (RT-PCR). Serum creatine kinase MB isoenzyme (CK-MB) and cardiac troponin I (cTnI) levels were analyzed by electrochemiluminescence. Apoptosis of human cardiac myocytes (HCM) was analyzed using fluorescence-activated cell sorting (FACS), and protein expression of caspase-3 was detected using Western blot.ResultsExpression levels of miRNA-21 in the serum of elderly patients with AMI were positively correlated with serum levels of CK-MB (r=0.3683, P=0.0229) and cTnI (r=0.5128, P=0.009). Following tumor necrosis factor (TNF)-α induction, the apoptosis rates of HCM transfected with the miRNA-21 mimic short hairpin RNA (shRNA) were downregulated by 39.1% compared with control HCM cells, and protein expression of c-Jun N-terminal kinases (JNK) and p38 were unchanged (P>0.05); protein expression of p-JNK, p-p38 and caspase-3 were downregulated by 37.1%, 35.8%, and 36.0%, respectively.ConclusionsExpression of miRNA-21 was upregulated in the serum of elderly patients with AMI, which inhibited TNF-α induced apoptosis in HCM by activating the JNK/p38/caspase-3 signaling pathway.

The inhibition of inducible nitric oxide synthase and oxidative stress by agmatine attenuates vascular dysfunction in rat acute endotoxemic model

Vascular dysfunction leading to hypotension is a major complication in patients with septic shock. Inducible nitric oxide synthase (iNOS) together with oxidative stress play an important role in development of vascular dysfunction in sepsis. Searching for an endogenous, safe and yet effective remedy was the chief goal for this study. The current study investigated the effect of agmatine (AGM), an endogenous metabolite of l-arginine, on sepsis-induced vascular dysfunction induced by lipopolysaccharides (LPS) in rats. AGM pretreatment (10mg/kg, i.v.) 1h before LPS (5mg/kg, i.v.) prevented the LPS-induced mortality and elevations in serum creatine kinase-MB isoenzyme (CK-MB) activity, lactate dehydrogenase (LDH) activity, C-reactive protein (CRP) level and total nitrite/nitrate (NOx) level after 24h from LPS injection. The elevation in aortic lipid peroxidation illustrated by increased malondialdehyde (MDA) content and the decrease in aortic glutathione (GSH) and superoxide dismutase (SOD) were also ameliorated by AGM. Additionally, AGM prevented LPS-induced elevation in mRNA expression of iNOS, while endothelial NOS (eNOS) mRNA was not affected. Furthermore AGM prevented the impaired aortic contraction to KCl and phenylephrine (PE) and endothelium-dependent relaxation to acetylcholine (ACh) without affecting endothelium-independent relaxation to sodium nitroprusside (SNP). In conclusion: AGM may represent a potential endogenous therapeutic candidate for sepsis-induced vascular dysfunction through its inhibiting effect on iNOS expression and oxidative stress.

Serum Creatine Kinase-MB Isoenzyme Activity among Subjects with Uncomplicated Essential Hypertension: Any Sex Differences.

Hypertension (high blood pressure) is a major health challenge and more women than men are affected by the condition. Complications as a result of this condition often lead to disabilities and premature death. The objective of this study was to evaluate creatine kinase-MB (CK-MB) activity in uncomplicated hypertension and to know whether sex differences exist in the activity of the enzyme. Serum creatine kinase-MB isoenzyme activity, troponin I, and lipid profile were evaluated in 140 male and 100 female Nigerians with hypertension. The control group was comprised of 100 (50 males and 50 females) normotensive subjects. Measured parameters were assayed using Selectra Pros chemistry analyzer. The means were compared between males and females using Students’t-test. The mean CK-MB activity of the female hypertensive subjects was significantly higher (p < 0.001) than the males. Similarly, the mean cardiac troponin I (cTnI) of the female hypertensive subjects was significantly higher (p < 0.001) than the males. Conversely, the mean CK-MB activity of the female normotensive subjects was significantly lower (p < 0.001) than the male counterparts. There was no difference in the levels of cTnI between male and female normotensive subjects. Serum CK-MB activity was higher in female than male hypertensive subjects. In the light of these results, cardiac markers should be routinely done in the evaluation of hypertensive subjects and sex-specific consideration may be recognized in the management of these patients.

Expression and location of HSP60 and HSP10 in the heart tissue of heat-stressed rats.

The present study aimed to analyze the expression levels and localizations of heat shock protein (HSP) 60 and HSP10 in the heart tissue of rats subjected to heat stress (42°C) for 0, 20, 80 and 100 min. Histopathological injuries and increased serum activities of serum lactate dehydrogenase and creatine kinase isoenzyme MB were detected in the heated rat myocardial cells. These results suggested that heat stress-induced acute degeneration may be sufficient to cause sudden death in animals by disrupting the function and permeability of the myocardial cell membrane. In addition, the expression levels of HSP60 were significantly increased following 20 min heat stress, whereas the expression levels of its cofactor HSP10 were not. Furthermore, the location of HSP60, but not of HSP10, was significantly altered during periods of heat stress. These results suggested that HSP60 in myocardial tissue may be more susceptive to the effects of heat stress as compared with HSP10, and that HSP10 is constitutively expressed in the heart of rats. The expression levels and localizations of HSP60 and HSP10 at the different time points of heat stress were not similar, which suggested that HSP60 and HSP10 may not form a complex in the heart tissue of heat-stressed rats.