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In this study, Lactobacillus paracasei subsp. paracasei NTU101 and Lactobacillus plantarum NTU 102 were used as starter to ferment soy-skim milk, and the optimal mixing ratio was evaluated. The influence of lactic acid bacteria (LAB)-fermented soy-skim milk on mucosal integrity in a gastric mucosal lesion rat model was also investigated. After 24 h, cell densities of L. paracasei subsp. paracasei NTU 101 and L. plantarum NTU 102 fermented in 75% soy milk and 25% milk (optimal condition) were 1.2 × 10(9) and 2.5 × 10(9) CFU/mL. After 180 days at 4 °C, the cell densities of the freeze-dried powders of the fermented soy-skim milks were 1 × 10(9) CFU/g; slight variations in pH and acidity were observed. Pylorus ligation with acidified ethanol treatment was used as the gastric lesion animal model. LAB-fermented soy-skim milk reduced the gastric lesion index and the lipid peroxides (LPO) of gastric mucosa and serum. Administration of the fermented soy-skim milk enhanced superoxide dismutase (SOD) activity and prostaglandin E(2) (PGE(2)) synthesis. Therefore, LAB-fermented soy-skim milk at 10(9) CFU/day protects against gastric injury.

Objective: To investigate the prevalence and correlation factors of carotid atherosclerosis (CAS) among workers of Chongqing Iron and Steel Corporation, and to offer information for the development of prevention program for cerebro-vascular disease., Methods: 512 asymptomatic individuals (mean age 56.8 years, ranged from 45 to 80) from the subsidiary workshop of Chongqing Iron and Steel Corporation were recruited. Demographic data and serum were collected and the internal-media thickness (IMT) of the CAS and the extent of plaque formation were evaluated by B-mode ultrasound. Logistic regression analysis was used to estimate the correlation factors of CAS. Results (1) The overall prevalence of carotid atherosclerosis among the iron and steel workers was 37.6%. IMT was found as 10.2% , with the prevalence rates of nonstenotic plaque and stenotic plaque were 25.0% and 2.4% respectively. (2) It was shown by logistic regression analysis that age (OR = 1.289, 95% CI: 1.014-1.568, P < 0.001), smoking (OR = 1.420, 95% CI: 0.802-3.872, P < 0.001), hypertension (OR= 4.530, 95% CI: 3.952-6.753, P < 0.001) , diabetes mellitus (OR = 2.285, 95% CI: 1.008-5.057, P < 0.001), HsCRP (OR = 1.273, 95% CI: 0.479-2.889, P = 0.037), TC (OR = 1.032, 95% CI: 0.320-1.882, P = 0.047) and LDL-C (OR = 2.313, 95% CI: 1.237-4.331, P = 0.008) were significantly associated with the severity of CAS. (3) Prevalence of the CAS increased with the increasing number of correlation factors., Conclusion: The prevalence of CAS among the iron and steel workers was higher than those in ordinary people of the same age. Age, smoking, hypertension, diabetes mellitus, HsCRP, TC and LDL-C were independent risk factors related to CAS. The prevalence of CAS was increasing parallel to the number of correlation factors.

We aimed to develop a double-injection model of intracerebral hemorrhage (ICH) in rabbits and to evaluate it as a tool for investigating post-ICH brain injury. Rabbits were injected with 300microL fresh autologous whole blood into the right basal ganglia. Behavioral changes were rated, brain water content (BWC) was measured and brain tissue morphology was also examined. ICH was established in 93.5% of the blood injection group. At 1, 3 and 7 days after ICH, there were significant differences in the total neurological scores (p<0.01) and BWC (p<0.01) between a sham-operated group and the ICH group. These findings suggest that the model produces a persistent neurological deficit, hematoma volume and perihematomal edema and closely mimics human hypertensive basal ganglia ICH; it is a controllable and reproducible hematoma that lends itself to quantitative investigation.

The title three compounds have been synthesized by solid state reactions at high temperatures, with excess indium as flux, and structurally characterized by single-crystal X-ray diffraction studies. Eu(3)Co(2)In(15) forms a new structure type and crystallizes in the tetragonal space group P4/mbm (No. 127), whereas KM(2)In(9) (M = Co, Ni) in the BaFe(2)Al(9) type crystallizes in the hexagonal space group P6/mmm (No. 191). Their structures all feature a three-dimensional anionic framework based on 1D [MIn(6)] single cluster chains composed of face-sharing [MIn(9)] clusters. In Eu(3)Co(2)In(15), two adjacent [CoIn(6)] single cluster chains form a [Co(2)In(11)] double cluster chain via corner-sharing In atoms as well as In-In bonds; the latter chains are further interconnected by additional indium atoms via In-In bonds into a complicated 3D framework, forming two types of tunnels along the c-axis, which are filled by the europium atoms. In KM(2)In(9), the [MIn(6)] single cluster chains are directly interconnected via corner sharing and exo In-In bonds into a 3D framework with the K(+) ions encapsulated in the 1D tunnels along the c-axis. Band structure calculations of three compounds based on density functional theory methods indicate that all three compounds are metallic.

Objective: To study the relationship between emotional, physical abuse and internet addiction disorder (IAD) among middle school students, in order to lay foundation for the development of prevention and control programs on IAD., Methods: Students selected from 76 classes in Grade One and Grade Two, filled out the anonymous questionnaire, which including demographic characteristics of students, Young's Internet Addiction Scale, Parent-Child Conflict Tactics Scales, etc. Non-conditional multivariate logistic regression model was employed to explore the relationship between child abuse and IAD. Independent variable was child abuse while dependent variable was IAD. Covariant variables would include demographic characteristics, as sex, age, style of household, et al., Results: Among 3798 students, the overall prevalence of emotional abuse was 81.1%, with 55.1% of them reported having experienced physical abuse. Among 3507 internet users, 5.2% was diagnosed as IAD. Results from the non-conditional multivariate logistic regression model showed that odds ratio of severe physical abuse and moderate physical abuse were 3.02 (1.63-5.58) and 4.00 (2.01-7.93)., Conclusion: Moderate physical abuse and severe physical abuse were possible risk factors of IAD.

HER2/neu oncogene-mediated malignancy is clearly associated with various human cancers. Therefore, HER2/neu targeting is an effective approach to cancer therapy. We have previously demonstrated that Epstein-Barr virus nuclear antigen-1 (EBNA1) can suppress HER2/neu oncogene expression, although EBNA1 itself has oncogenic potential. Here, we found that the N-terminal domain of EBNA1 alone, named EBNA1-NT, which contains the N-terminal region of amino acid residues 1-86 of EBNA1, is required and sufficient to suppress HER2/neu oncogene expression at the transcriptional level. Furthermore, in EBNA1-NT-transfected HER2/neu-overexpressing cells, we found EBNA1-NT could down-regulate the endogenous production of p185(HER2/neu), lower transformation ability, sensitize paclitaxel-induced apoptosis and decrease tumorigenic potential. These data suggest that EBNA1-NT may act as a repressor of the HER2/neu oncogene.

Objective: To determine the contents of NOS and endogenous CO in the penile corpus cavernosum of Wistar rat models of hyperthyroid and hypothyroid, to detect the effects of thyroid hormone and endogenous CO on rat penile erection, and to further investigate the influence of thyroid hormone on human penile erection., Methods: Fifty 3-month-old male Wistar rats were randomly and equally divided into 5 groups: a hyperthyroid, a hyperthyroid treatment, a hypothyroid, a hypothyroid treatment and a control group. The contents of NOS and CO were detected by ultraviolet spectrophotometry., Results: Compared with the controls, the content of NOS was significantly decreased in both the hyperthyroid and the hypothyroid groups (P < 0.01), even lower in the latter than in the former (P < 0.01), and so was the content of CO (P < 0.01), lower in the former than in the latter (P < 0.01). After treatment, the levels of NOS and CO were both elevated in either the hyperthyroid or the hypothyroid group, with no significant difference from that of the controls (P > 0.05)., Conclusion: The contents of NOS and CO in the penile corpus cavernosum decrease with the imbalance of thyroid hormones, but return to normal once the imbalance is corrected. Under the same conditions, hypothyroid inflicts more damage on penile erection than hyperthyroid.

Objective: To investigate the predictors of road traffic accidents (RTAs) in middle school students, a case-control study was designed to seek new clues for prevention and control on RTAs in this population., Methods: Cases were 116 students who experienced RTAs from 2005/01/01 to 2005/12/31. 116 students were served as controls who were from the same class of the cases and matched by age and sex. Information on personal behavior, family conditions, and knowledge, attitude, practice (KAB) on road traffic of these children were collected and compared. Conditional logistic regressions were conducted to analyze the relationship between these factors and RTAs., Results: Compared to the controls, the cases showed more negative attitude and risk-taking-behaviors especially in riding bicycle. However, knowledge on road safety showed less significant differences between case and control groups. The negative attitude and risk-taking-behaviors were independent risk factors for RTAs by logistic regression. In addition, distance from home to school that longer than 1000 m seemed a risk factors for RTAs, while high education levels among mothers was a protective factor., Conclusion: Road safety attitude and behavior, distance from home to school were the main influence factors for RTAs among middle school students. It seemed more important in improving the attitude and behavior on road safety than to enhance the knowledge for prevention and control of RTAs among middle school students.

Objective: To detect the levels of nitric oxide synthetase (NOS) and carbon monoxide (CO) in the penile corpus cavernous of adult male Wistar rats with high homocysteine (Hhcy) and to explore the relationship of NOS and CO levels with erectile dysfunction., Methods: Twenty Wistar rats were equally and randomly divided into a control and an Hhcy group and fed on normal diet and normal diet with 3.0% methionine respectively. Four weeks later, the levels of NOS and CO in the penile corpus cavernous were detected by ultraviolet spectrophotometry and that of serum homocysteine by the cycle enzyme method., Results: Compared with the control group, the levels of NOS and CO in the penile corpus cavernous were significantly lower in the Hhcy group, (6.45 +/- 1.12) nmol/(g x min) vs (10.77 +/- 0.60) nmol/(g x min) and (10.60 +/- 0.92) micromol/L vs (13.36 +/- 0.44) micromol/L, while that of homocysteine was significantly higher, (22.32 +/- 1.65) micromol/L) vs (4.90 +/- 1.73) micromol/L., Conclusion: Four-week diet with methionine can cause Hhcy and significantly decreased levels of NOS and CO in the penile corpus cavernous in Wistar rats. Hhcy is an independent risk factor of erectile dysfunction.

Previously, we reported that apoptosis of cerebellar granular neurons induced by low-K+ and serum-free (LK-S) was associated with an increase in the A-type K+ channel current (I(A)), and an elevated expression of main alpha-subunit of the I(A) channel, which is known as Kv4.2 and Kv4.3. Here, we show, as assessed by quantitative RT-PCR and whole-cell recording, that besides Kv4.2 and Kv4.3, Kv1.1 is very important for I(A) channel. The expression of Kv1.1 was elevated in the apoptotic neurons, while silencing Kv1.1 expression by siRNA reduced the I(A) amplitude of the apoptotic neuron, and increased neuron viability. Inhibiting Kv1.1 current by dendrotoxin-K evoked a similar effect of reduction of I(A) amplitude and protection of neurons. Applying a protein kinase C (PKC) activator, phorbol ester acetate A (PMA) mimicked the LK-S-induced neuronal apoptotic effect, enhanced the I(A) amplitude and reduced the granule cell viability. The PKC inhibitor, bisindolylmaleimide I and Gö6976 protected the cell against apoptosis induced by LK-S. After silencing the Kv1.1 gene, the effect of PMA on the residual K+ current was reduced significantly. Quantitative RT-PCR and Western immunoblot techniques revealed that LK-S treatment and PMA increased the level of the expression of Kv1.1, in contrast, bisindolylmaleimide I inhibited Kv1.1 expression. In addition, the activation of the PKC isoform was identified in apoptotic neurons. We thus conclude that in the rat cerebellar granule cell, the I(A) channel associated with apoptotic neurons is encoded mainly by the Kv1.1 gene, and that the PKC pathway promotes neuronal apoptosis by a brief modulation of the I(A) amplitude and a permanent increase in the levels of expression of the Kv1.1 alpha-subunit.

The immunological mechanisms on adventitial inflammation has received much attention, while the contribution of nerves to adventitial inflammation has largely been ignored. Although the mechanism of initial chemotaxis of the adventitial inflammatory cells remains unknown, vascular nerves were frequently found in the inflammatory lesions of coronary adventitia and adventitial mast cells connect with sensory nerve fibers in atherosclerotic coronary arteries. The sensory nerves in contact with adventitial mast cells contained the neuropeptides SP and CGRP. These neuropeptides play an important role in the amplification of tissue injury by the increase of both vascular permeability and neutrophil recruitment, and the term ''neurogenic inflammation'' has been coined. Activation of adventitial mast cells, with ensuing release of vasoactive compounds, may cause vasoconstriction in atherosclerotic coronary segments. Therefore, we hypothesize that adventitial vanilloid receptor TRPV1 and sensory C-fibers may play a pistol role for adventitial inflammation.

Attractin (ATRN) and Attractin-like 1 (ATRNL1) are highly similar type I transmembrane proteins. Atrn null mutant mice have a pleiotropic phenotype including dark fur, juvenile-onset spongiform neurodegeneration, hypomyelination, tremor, and reduced body weight and adiposity, implicating ATRN in numerous biological processes. Bioinformatic analysis indicated that Atrn and Atrnl1 arose from a common ancestral gene early in vertebrate evolution. To investigate the genetics of the ATRN system and explore potential redundancy between Atrn and Atrnl1, we generated and characterized Atrnl1 loss- and gain-of-function mutations in mice. Atrnl1 mutant mice were grossly normal with no alterations of pigmentation, central nervous system pathology or body weight. Atrn null mutant mice carrying a beta-actin promoter-driven Atrnl1 transgene had normal, agouti-banded hairs and significantly delayed onset of spongiform neurodegeneration, indicating that over-expression of ATRNL1 compensates for loss of ATRN. Thus, the two genes are redundant from the perspective of gain-of-function but not loss-of-function mutations., ((c) 2007 Wiley-Liss, Inc.)

Objective: To determine whether measurement of cardiac troponin T (cTnT) concentration in newly diagnosed peripartum cardiomyopathy (PPCM) can be used to predict persistent left ventricular dysfunction after a 6-month follow-up., Patients and Methods: This was a prospective, multiple-centre clinical trial that studied 106 patients with newly diagnosed PPCM surviving over 6 months. cTnT concentration was measured within 2 weeks of the onset of PPCM., Results: Serum cTnT concentration was negatively correlated with left ventricular ejection fraction (LVEF) at follow-up (LVEF, r = -0.518, p = 0.0001). Analysis by receiver operator characteristic curve yielded an area under the curve of 0.764 (95% CI 0.669 to 0.860, p = 0.0001, vs null hypothesis value 0.5) for cTnT, and a cTnT concentration cut off of >0.04 ng/ml, predicting persistent left ventricular dysfunction with a sensitivity of 54.9% and a specificity of 90.9%. Among 106 recruited patients, there were 33 patients with cTnT concentrations >0.04 ng/ml and 73 patients with cTnT concentrations < or =0.04 ng/ml. After a 6-month follow-up, there was significantly smaller LVEF (35.42% (13.04% vs 50.16% (10.48%, p = 0.0001) and more persistent left ventricular dysfunction (84.8% vs 31.5%, OR = 12.17 (95% CI 4.17 to 35.57), p = 0.001) in patients with cTnT >0.04 ng/ml than in patients with cTnT < or =0.04 ng/ml., Conclusion: Serum cTnT concentration measured within 2 weeks of the onset of PPCM was correlated negatively with LVEF at follow-up. This marker offers a simple, quick, inexpensive, non-invasive method for predicting a persistent LVEF of < or =50%. A cTnT concentration of >0.04 ng/ml predicted persistent left ventricular dysfunction with a sensitivity of 54.9% and a specificity of 90.9%.

The article entitled, "Cardiac Troponin T: An Early Molecule Marker of Normalization of Left Ventricular Ejection Fraction in Patients with Peripartum Cardiomyopathy", by Li et al, which originally was published in this space, has been removed because an article by the same authors and reporting very similar work already has been published in HEART [Published Online First: 25 October 2006], entitled, "Troponin T measurement can predict persistent left ventricular dysfunction in peripartum cardiomyopathy", by Hu et al. The printed version of the article in HEART can be found at Heart 2007;93:488-490.

Objective: To study the effects of lutein on apoptosis and its mechanism., Method: The cells of human esophageal carcinoma EC9706 were grown in RPMI medium containing 10% bovine serum and were treated with lutein at 100 microg x mL(-1) concentration. Flow cytometry was employed to investigate the effects of lutein on cell apoptosis of EC9706 cells. Histochemistry was performed to determine apoptosis-related protein expresion., Result: Flow cytometry analyses revealed that lutein increased EC9706 cell apoptosis ratio when treated with lutein 100 microg x mL(-1) at 96 h. Lutein decreased the expression of Bcl-2 protein and increased the expression of Bax protein in EC9706 cells., Conclusion: Lutein could inhibit mitosis and stimulate apoptosis of EC9706 cells. The apoptotic effect may result from the down-regulation of expression of Bcl-2 and up-regulation expression of Bax.

A variety of cells, including fibroblasts, mast cells, macrophages, and ganglionic cells, are present in coronary artery adventitia. In the infarct-related coronary arteries of myocardial infarction patients, the majority of mast cells are found in the outer layer of the adventitia. Neurogenic stimulation of mast cells in the adventitia of coronary arteries may release vasoactive compounds, such as histamine and leukotrienes, which can contribute to the complex neurohormonal response that leads to abnormal coronary vasoconstriction. Lymphocytes and bacteria are also present mainly in the adventitial layer. Chlamydia pneumoniae is directly involved in the development of adventitial and plaque inflammation (pan-arteritis), leading to plaque rupture. Adventitial O(2)(-) may also play an extensive role in the control of vascular tone. Therefore, adventitial inflammation may play a pivotal role for atherosclerotic lesion development and atheroma instability.

Recent years have seen an enormous amount of experimental and clinical research into role of the pulmonary veins (PVs) in atrial fibrillation (AF). The PVs contain cardiomyocytes with easily inducible arrhythmogenic activity due to the enhanced automaticity, induction of triggered activity, and genesis of microreentrant circuits. The enhanced automaticity, induced triggered activity, either alone or in combination with the reentrant mechanisms, may play a role in the initiation of PVs AF. Detailed mapping studies suggest that reentry within the PVs is most likely responsible for their arrhythmogenicity. There is no doubt that the PVs represent the most important source of arrhythmogenic activity in patients with paroxysmal AF. In AF patients with risk factors for development of AF, the presence of the pathological situation is important in enhancing the PV arrhythmogenic activity. Coronary sinus or superior vena cava may also be a source of rapid repetitive electrical activity during AF. Thus, AF should be considered a kind of vascular disease. Moreover, in patients with paroxysmal AF originating from the PVs, a wide spectrum of atrial arrhythmias may coexist, including paroxysms of atrial premature, tachycardia, flutter and fibrillation. This kind of arrhythmias should be named as PV atrial arrhythmias. These new views will help understand the mechanism, diagnosis and treatment method for AF.

Atrial fibrillation (AF) is not a benign condition. Although AF is the most common sustained cardiac arrhythmia, the management of AF remains controversial. In recent years, several clinical trials comparing rhythm control with rate control for AF management found that rhythm control offers no advantage over rate control, but the findings may not apply equally to all patients. The main point is the high risk of AF recurrence in these trials and the toxic effects and poor efficacy of presently available antiarrhythmic drugs negate the benefit of rhythm control. Rhythm control should be considered as the preferred initial therapy if sinus rhythm had been maintained in a higher proportion of patients. For the moment, the optimal strategy to treat AF should be individualized, with a bias towards rate control if patients are at high risk for arrhythmias or can be kept asymptomatic. Rhythm control may still be justified in younger patients or patients who still have symptoms despite adequate rate control. In addition, patients with new or first-episode AF often warrant at least an initial trial of rhythm control.

Calcific aortic valve disease is common among the elderly. Until recently, the concept that calcific aortic valve disease is a degenerative and unmodifiable process basically induced by long-lasting mechanical stress was generally accepted. However calcific aortic valve disease is not merely related to age-associated "wear and tear". The development and progression of calcific aortic valve disease are based on an active process that shares a number of similarities with atherosclerosis. Statins and angiotensin-converting enzyme inhibitors have been shown to slow calcium accumulation in aortic valves. Thus, calcific aortic valve disease should not be considered as a degenerative disease anymore.

Compelling evidence indicates that excessive potassium (K+) efflux and intracellular K+ depletion are the key early steps in apoptosis. Previously, we reported that apoptosis of cerebellar granule neurons induced by incubation in low-K+ (5 mM) and serum-free medium was associated with an increase in A-type transient inactivation of K+ channel current (IA) amplitude and modulation of channels' gating properties. Here, we showed that a classic K+ channel blocker, 4-aminopyradine (4-AP), significantly inhibited IA amplitude in a concentration-dependent manner (reduction of current by 10 microM and 10 mM 4-AP was 11.4+/-1.3% and 72.2+/-3.3%, respectively). Moreover, 4-AP modified the steady-state activation and inactivation kinetics of IA channels, such that the activation and inactivation curves were shifted to the right about 20 mV and 17 mV, respectively. Fluorescence staining showed that 4-AP dramatically increased the viability of cells undergoing apoptosis in a dose-dependent manner. That is, while 5 mM 4-AP was present, cell viability was 84.9+/-5.2%. Consistent with the cell viability analysis, internucleosomal DNA fragmentation by gel electrophoresis analysis showed that 5 mM 4-AP also protected against neuronal apoptosis. Furthermore, 4-AP significantly inhibited cytochrome c release and caspase-3 activity induced by low-K+/serum-free incubation. Finally, current-clamp analysis indicated that 5 mM 4-AP did not significantly depolarize the membrane potential. These results suggest that 4-AP has robust neuroprotective effects on apoptotic granule cells. The neuroprotective effect of 4-AP is likely not due to membrane depolarization, but rather that 4-AP may modulate the gating properties of IA channels in an anti-apoptotic manner.

Objective: To compare rate control and rhythm control strategies in patients with atrial fibrillation (AF) after percutaneous mitral balloon valvotomy (PMV)., Methods: 183 patients with AF after successful PMV, with AF duration

Objective: C-reactive protein (CRP) is considered a risk factor for coronary artery disease. In addition to its lipid-lowering properties, statin decreases the level of CRP. Abrupt cessation of statin therapy during treatment could increase CRP level independently of the elevation of serum lipids and the incidence of cardiac events in patients with atherosclerotic heart disease. Xuezhikang (XZK), an extract of cholestin, has a marked modulating effect on lipid and CRP concentrations in different study time course. However, no attention has been paid to the changes of lipid profile and CRP concentrations after withdrawal of XZK treatment. This study was designed to explore short-term time course effects on lipid profile and CRP concentrations after withdrawal of XZK treatment in coronary heart disease patients., Materials and Methods: Seventy-five consecutive patients with documented coronary heart disease were randomly divided into three groups: 1. Pretreatment with XZK 1,200 mg daily for 6 weeks and then replaced by placebo (XZK discontinued group; n = 25); 2. Treatment with XZK 1,200 mg daily throughout the study (XZK continued group; n = 25); or 3. Placebo (no XZK group; n = 25). Lipid levels (total cholesterol, HDL-C, LDL-C and triglycerides) and CRP were assessed before receiving the XZK therapy, 1 day before discontinuation of XZK, and on days 1, 2, 3, 7 and 14 after discontinuation of XZK, respectively., Results: After 6-week XZK treatment, the fasting total cholesterol, LDL-C, triglyceride and median hs-CRP concentrations decreased, whereas HDL-C concentration increased significantly (p < 0.001, respectively). At day 14 after discontinuation of XZK therapy, total cholesterol (15%), LDL-C (17%) and triglyceride (20%) significantly increased (p < 0.001, respectively), whereas HDL-C level (15%) significantly decreased (p < 0.05). The median level of CRP increased by 11, 65, 128, 103 and 101% on the first, second, third, seventh, and fourteenth day after withdrawal of XZK therapy (p > 0.05, <0.05, <0.001, <0.001, <0.001, compared with 1 day before withdrawal of XZK therapy, respectively). There was a prominent rebound of CRP concentration 3 days after discontinuation of XZK therapy. At this time point, hs-CRP concentration was higher than in the placebo group (p < 0.05). Seven to 14 days after discontinuation of XZK therapy, the hs-CRP concentration declined to a similar level as in the placebo group. No significant correlation was seen between the changes in hs-CRP and lipid profile at all time points., Conclusions: The level of hs-CRP increases on the second day after withdrawal of XZK and there is a prominent rebound 3 days after discontinuation of XZK therapy. The increase of CRP ends within 7 days, where lipids increase at 14 days after discontinuation of XZK therapy. The results may be clinically important for patients with coronary artery disease.

Objective: To construct a recombinant adenoviral vector carrying HIF-1alpha gene and explore the therapeutic effect of HIF-1alpha on focal cerebral ischemia in adult rats., Methods: The AdEasy System was used to construct the recombinant adenoviral vector carrying HIF-1alpha gene and green fluorescent protein and PCR was used to identify the HIF-1alpha gene. Middle cerebral artery occlusion (MCAo) and reperfusion models were established and divided into Ad-HIF-1alpha group, Ad group and NS group. After Ad-HIF-1alpha, Ad and NS were injected into the ischemic ventricle, expression of Ad-HIF-1alpha was observed and its therapeutic effect was evaluated by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining and neurological severity scores., Results: GFP expression distributed apart from the ventricle and reached a peak at 14 days and persisted for about 4 weeks under fluorescent microscope. The neurological severity scores was 2.4 +/- 0.5 at 24 h in Ad-HIF-1alpha group and there was no statistical significance compared with Ad group (2.6 +/- 0.5) and NS group (2.7 +/- 0.7) (P > 0.05). The scores were 1.6 +/- 0.7 at 48 h and 0.9 +/- 0.6 at 72 h in Ad-HIF-1alpha group, and there were statistical significance compared with Ad group (2.9 +/- 0.6 and 3.2 +/- 0.6 respectively) and NS group (3.0 +/- 0.7 and 3.2 +/- 0.8) (P < 0.05). The infarct volume was 81.2 mm(3) +/- 1.4 mm(3) at 72 h in Ad-HIF-1alpha group and there was statistical significance compared with Ad group (173.9 mm(3) +/- 1.3 mm(3)) and NS group (171.7 mm(3) +/- 6.2 mm(3)) (P < 0.05)., Conclusion: HIF-1alpha gene had definite therapeutic effect on focal cerebral ischemia in adult rats, which settles a foundation for next HIF-1alpha gene study and clinic application.

Objective: To study the effect of Shuxuetong injection on cerebral tissue and brain microvascular endothelial cell(BMEC) in secreting tissue-type plasminogen activator (tPA)., Method: The experiment in vivo was designed for three groups: sham-operation group, focal cerebral ischemia model group, Shuxuetong injection-treated group. The gene expression and the activity of tPA on brains were detected by RT-PCR and chromogenic substrate analysis respectively after treatment for five days; The experiment in vitro: The cultured BMEC were divided into experimental group and control group. The Shuxuetong injection were added into experimental group and the BMECs in the experimatal groups were treated for 24 h. The gene expression and the activity of tPA on cultured BMEC were detected by RT-PCR and chromogenic substrate analysis respectively., Result: The gene expression and the content of tPA on both cerebral tissue and BMEC in the experimatal groups were significantly higher than that in the control groups., Conclusion: Shuxuetong injection could accelerate secretion of tPA, and therefore enhance fibrinolysis.

Experimental and clinical findings support the essential role of interleukin (IL)-6 in the pathogenesis of various human cancers and provide a rationale for targeted therapeutic investigations. A novel peptide, S7, which selectively binds to IL-6 receptor (IL-6R) alpha chain (gp80) and broadly inhibits IL-6-mediated events, was identified using phage display library screening. The synthetic S7 peptide specifically bound to soluble IL-6R as well as cognate human IL-6R alpha, resulting in a dose-dependent blockade of the interaction between IL-6 and IL-6R alpha. S7 peptide prevents IL-6-mediated survival signaling and sensitizes cervical cancer cells to chemotherapeutic compounds in vitro. The in vitro analysis of antiangiogenic activity showed that S7 peptide substantially inhibits IL-6-induced vascular endothelial growth factor-A expression and angiogenesis in different cancer cell lines. Furthermore, S7 peptide was bioavailable in vivo, leading to a significant suppression of IL-6-induced vascular endothelial growth factor-mediated cervical tumor growth in severe combined immunodeficient mice. These observations show the feasibility of targeting IL-6/IL-6R interaction using the small peptide and highlight its potential in the clinical applications.

Compelling evidence indicates that excessive K+ efflux and intracellular K+ depletion are key early steps in apoptosis. Previously, we reported that apoptosis of cerebellar granular neurons induced by incubation under low K+ (5 mM) conditions was associated with an increase in delayed rectifier outward K+ current (IK) amplitude and caspase-3 activity. Moreover, the melatonin receptor antagonist 4P-PDOT abrogated the effects of 2-iodomelatonin on IK augmentation, caspase-3 activity and apoptosis. Here, we show that incubation under low K+/serum-free conditions for 6 hr led to a dramatic increase in the A-type transient outward K+ current (IA) (a 27% increase; n=31); in addition, fluorescence staining showed that under these conditions, cell viability decreased by 30% compared with the control. Treatment with 2-iodomelatonin inhibited the IA amplitude recorded from control and apoptotic cells in a concentration-dependent manner and modified the IA channel activation kinetics of cells under control conditions. Moreover, 2-iodomelatonin increased the viability of cell undergoing apoptosis. Interestingly, 4P-PDOT did not abrogate the effect of 2-iodomelatonin on IA augmentation under these conditions; in the presence of 4P-PDOT (100 microm), 2-iodomelatonin reduced the average IA by 41+/-4%, which was similar to the effect of 2-iodomelatonin alone. These results suggest that the neuroprotective effects of 2-idomelatonin are not only because of its antioxidant or receptor-activating properties, but rather that 2-iodomelatonin may inhibit IA channels by acting as a channel blocker.

Cell migration is mediated by ion channels and transporters, and plays crucial roles in a variety of physiological and pathological processes. Previously, our studies have shown that a Ca(2+)-regulated K(+) current exists in B-16 murine melanoma cells, and that endothelin-1 (ET-1) inhibits the K(+) current via a PKC-dependent pathway. In the present study, patch-clamp whole-cell recording and transwell migration assays were used to examine the effects of ET-1 on B-16 murine melanoma cell migration. ET-1 (100 nM in the injection pipette and 10 nM in the incubation medium) decreased the K(+) current amplitude by 33.0 +/- 2.5% and inhibited migration of B-16 cells by 57.4 +/- 9.4%. Similarly, the Ca(2+)-regulated K(+) channel blockers, BaCl(2) and quinidine, decreased the K(+) current by 20.5 +/- 1.0% and 36.6 +/- 1.2%, respectively, and slowed migration of B-16 melanoma cells by 37.1 +/- 8.6% and 42.7 +/- 8.8%, respectively. The effect of ET-1 on the K(+) current and cell migration was simulated by ET-3. In contrast, the K(+) channel opener, diclofenac, increased the K(+) current by 128.8 +/- 11.7%, 257.4 +/- 35.8% at concentrations of 1 and 5 mM, respectively. Likewise, the migration of B-16 murine melanoma cells dramatically increased by 75.6 +/- 12.7% in the presence of 100 microM diclofenac in incubation medium. Furthermore, the ET-1- and ET-3-induced inhibition of K(+) current and migration was abrogated by diclofenac. In the presence of diclofenac, ET-1 only reduced the K(+) current amplitude by 10.6 +/- 1.1%, and slowed B-16 cell migration by only 10.8 +/- 8.9%. The results suggest that the K(+) channel-dependent migration of B-16 melanoma cells is modulated by ET-1. Cell Motil., (Copyright 2004 Wiley-Liss, Inc.)

Activation of K(+) current plays a critical role in the control of programmed cell death. In the present study, whole-cell patch-clamp recording, a caspase-3 activity assay, and flow cytometric analysis were used to examine the effects of the MT2 melatonin receptor agonist 2-iodomelatonin on the delayed-rectifier K(+) current (IK) and the prevention of apoptosis. It was found that apoptosis of cerebellar granular neurons induced by low-K(+) (5 mm) incubation was associated with an increase in IK amplitude and caspase-3 activity. After 6 hr of low-K(+) treatment, IK was increased by 45% (n = 86). Flow cytometry showed that the apoptosis rate increased by 333% compared with the control neurons. In addition, exposure of cultured granule cells to low K(+) also resulted in a significant activation of caspase-3, by 466%. 2-Iodomelatonin (10 microm in injection pipette) inhibited the IK amplitude recorded from control cells and from cells undergoing apoptosis. However, 2-iodomelatonin only modified the IK-channel activation kinetics of cells under both conditions. Furthermore, 2-iodomelatonin reduced the rate of apoptosis and caspase-3 activation, by 66 and 64%, respectively. The melatonin receptor antagonist, 4P-PDOT, abrogated the effect of 2-iodomelatonin on the IK augmentation, caspase-3 activity, and apoptosis. These results suggest that the neuroprotective effects of melatonin are not only because of its function as a powerful antioxidant, but also to its interactions with specific receptors. The effect of 2-iodomelatonin against apoptosis may be mediated by activating a melatonin receptor, which modulates IK channels and reduces K(+) efflux.

Ovarian follicle development is dependent on growth factors that stimulate cell proliferation and act as survival factors to prevent apoptosis of follicle cells. We examined the mechanism of the protective effect of IGF-I against Fas ligand-induced apoptosis of granulosa cells and its relationship to cell proliferation. IGF-I activated both the phosphoinositide 3'-OH kinase (PI3K) and the MAPK pathways. Experiments using specific inhibitors of these pathways showed that protection by IGF-I was mediated by the PI3K pathway and not the MAPK pathway. Recombinant adenoviruses were used to test whether the downstream target of PI3K activation, Akt kinase, was required for protection against apoptosis. Expression of dominant negative Akt prevented protection by IGF-I whereas expression of constitutively active Akt (myrAkt) mimicked the effect of IGF-I. Treatment with IGF-I, or expression of myrAkt, increased progression from G(0)/G(1) to S phase of the cell cycle whereas expression of dominant negative Akt inhibited G(0)/G(1) to S phase progression and prevented the stimulatory effect of IGF-I. We tested whether cell cycle progression was required for protection from apoptosis using the cyclin-dependent kinase-2 inhibitor roscovitine, which blocks cells at the G(1)/S transition. Roscovitine prevented the protective effect of IGF-I and myrAkt expression against apoptosis. Therefore, activation of Akt is not sufficient to protect granulosa cells from apoptosis in the absence of cell cycle progression. In summary, IGF-I protects granulosa cells from apoptosis by activation of the PI3K/Akt pathway. This protective effect can occur only when progression from G(1) to S phase of the cell cycle regulated by the PI3K/Akt pathway is unperturbed.

Growth factors and steroids play an important role in the regulation of ovarian follicular development. In cattle, two of the earliest detectable differences between the healthy dominant follicle selected for development to the ovulatory stage and subordinate follicles destined to undergo atresia are the greater availability of IGF and the greater capacity to produce estradiol in the dominant follicle. We have shown that IGF-I and estradiol stimulate the proliferation of bovine granulosa cells in vitro and promote granulosa cell survival by increasing resistance to apoptosis. Furthermore, the ability of IGF-I and estradiol to increase resistance to apoptosis is tied to their ability to promote progression through the cell cycle. Blocking the cell cycle at the transition between the first gap phase and the DNA synthesis phase using a specific inhibitor prevented the protective effects of IGF-I and estradiol against apoptosis. Further experiments showed that the protective effect of IGF-I against apoptosis is mediated by the stimulation of phosphatidylinositol 3-kinase and its downstream target, protein kinase B/Akt. Constitutive activation of Akt by the infection of granulosa cells with a recombinant Akt adenovirus protected against apoptosis, and this effect also depended on cell cycle progression. These experiments show that the protective effect of estradiol and IGF-I against apoptosis depends on unperturbed progression through the cell cycle. Once follicles have developed to the preovulatory stage, the LH surge induces terminal differentiation of granulosa cells and withdrawal from the cell cycle. Bovine granulosa cells withdraw from the cell cycle by 12 h after the LH surge and become resistant to apoptosis, even in the absence of growth factors. Treatment with a progesterone receptor antagonist in vitro caused reentry of granulosa cells into the cell cycle and susceptibility to apoptosis, suggesting that induction of progesterone receptor expression by the LH surge is required for cell cycle withdrawal and resistance to apoptosis. In summary, the susceptibility of granulosa cells to apoptosis depends on the cell cycle. Proliferating granulosa cells in growing follicles depend on growth factors for survival, whereas cells that have terminally differentiated in response to the LH surge are resistant to apoptosis and relatively independent of growth factors for survival.

Objective: To study the accumulation of fluoride in rat hippocampus and its effect on cholinesterase activity., Methods: Rats were subchronically exposed to NaF, and fluoride concentration and cholinesterase activity in rat hippocampus were determined., Results: Fluoride concentration in rat hippocampus was significantly correlated with the dosage of fluoride, and there were significant differences among high dosage group [(13.03 +/- 1.79) micro g/g], low dosage group [(9.83 +/- 0.92) micro g/g] and control [(8.27 +/- 1.11) micro g/g], P < 0.01. Acetylcholinesterase activities among three groups [(0.111 +/- 0.031) micro mol/mg, (0.143 +/- 0.025) micro mol/mg, (0.183 +/- 0.027) micro mol/mg] were also significantly different (P < 0.01), which was negatively correlated with fluoride concentration in rat hippocampus (r = -0.700, P < 0.01). The activity of butylcholinesterase in high dosage group [(0.041 +/- 0.010) micro mol/mg] was different from that of control [(0.067 +/- 0.025) micro mol/mg, P < 0.05], but the activity was not significantly related with fluoride concentration in rat hippocampus (r = -0.317, P = 0.094)., Conclusion: Fluoride may go through the blood-brain barrier and accumulate in rat hippocampus, and inhibit the activity of cholinesterase.

We had previously found that co-injection of dextromethorphan, an antitussive drug and a non-competitive NMDA receptor antagonist, with morphine into dam rats throughout the pregnancy period could attenuate the naloxone-precipitated morphine withdrawal syndrome in their offspring. In the present study, we further tested whether postnatal injection of dextromethorphan into the neonatal rats or a 3-day co-injection of dextromethorphan with morphine into the dam rats before delivery is also effective. Female Sprague-Dawley rats were bi-daily injected with escalating doses of morphine from a week before mating till the first postnatal week. Withdrawal syndrome of morphine in the offspring, manifested mainly as abdominal stretching, was generated by injection of naloxone on postnatal day 5. Direct injection of dextromethorphan into the offspring effectively reduced the severity of naloxone-precipitated abdominal stretching in a dose-dependent manner. A 3-day co-treatment with dextromethorphan given to the dam rat before delivery also had a similar attenuating effect, but the efficacy was lower than that produced by postnatal injection. Thus, the results from the present study support that dextromethorphan is of potential in treating or preventing neonatal morphine withdrawal syndrome.

The purpose of this study was to determine whether 0.8-1 mA, 2 Hz of percutaneous electrical stimulation could affect the regeneration of a 10-mm gap of rat sciatic nerve created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes. Six weeks after implantation, though the group receiving the electrical stimulation had a lower success percentage of regeneration (57%) compared with the controls receiving no stimulation (70%), quantitative histology of the successfully regenerated nerves revealed that the mean values of the axon density, blood vessel number, blood vessel area, and percentage of blood vessel area in total nerve area in the group with the electrical stimulation were all significantly larger than those in the controls (p < 0.05). These results showed that the electrical stimulation could elicit rehabilitating effects on the regenerated nerves., (Copyright 2001 John Wiley & Sons, Inc. J Biomed Mater Res 57: 541-549, 2001)

Ovarian follicular atresia occurs by apoptosis of granulosa and theca cells. The Fas antigen (Fas), a cell surface receptor that triggers apoptosis when activated by Fas ligand (FasL), may be involved in this process. A possible role of the Fas pathway in mediating serum withdrawal-induced apoptosis of granulosa cells was examined. Granulosa cells collected from 5- to 10-mm bovine follicles were cultured in DMEM-F12 containing serum for 3 days, deprived of serum, and live cells were counted at various times after serum withdrawal. Cell death increased significantly 6 h after serum withdrawal (21% +/- 7%; P: < 0.05 vs. 0 h) and continued to increase until 24 h (43% +/- 6%). No further increases in cell death were observed through 72 h. Detection of the translocation of phosphatidylserine to the outer surface of the cell membrane by annexin V binding indicated that cells died by apoptosis. Quantitative reverse transcriptase-polymerase chain reaction assays showed no changes in Fas mRNA levels but a 4.7-fold increase in FasL mRNA 3 h after serum withdrawal (P: < 0.05 vs. 0 h). FasL mRNA remained elevated through 24 h and returned to basal levels at 48 h. Immunohistochemical staining showed that both Fas and FasL protein increased on the cell surface within 3 h and remained elevated through 12 h (the last time point tested). Binding of FasL to Fas was blocked with two reagents that bind to the extracellular domain of FasL: an anti-FasL antibody and Fas:Fc, a chimeric protein consisting of the Fc portion of human immunoglobulin G and the extracellular domain of human Fas. Cell death 24 h after serum withdrawal was reduced 55% +/- 10% and 34% +/- 12% by anti-FasL antibody and Fas:Fc, respectively (P: < 0.05 vs. no blocking protein). In conclusion, serum withdrawal-induced apoptosis of bovine granulosa cells is mediated at least partially by Fas/FasL interactions. These results are consistent with a potential role of Fas in an autocrine or paracrine pathway to trigger ovarian follicular atresia.

After the master cell stock(mcs) and working cell bank of more than 30 different strains of 7 animal kidney cell lines (F-81 or CRFK cell line, MDCK cell line, Vero or Vero-2 cell line, MA-104 cell line and BHK-21 cell line) were established in China, the chromosomal number variations and structural aberrations of the above lines, primary feline or canine kidney cell (FKC or CKC) and HeLa cell line were investigated and their karyotypes of routine or Giemsa chromosomal bands were analyzed. The carcinogenesis or tumorigenicity testing of these cells in about 700 nude mice and for colony formation in soft agar (SA) and for agglutination under different concentration of plant lectins was carried out. Both tumorigenicity-negative strains of F-81, CRFK, Vero or Vero-2 lines and very-low-tumorigenicity strains of MDCK line were successfully selected and evaluated for the production of canine or feline combination viral vaccines, which are free of infectious agents, and described with respect to cytogenetic characteristics and tumorigenicity. Rate of modal chromosome number represents the ratio of cell number having modal chromosome number to all the split cell number analyzed at random. Rate of difference represents the ratio of difference of the rate of modal chromosome number between mcs (master cell stock) + n and mcs passages. The chromosomal analysis results showed that the ratio of difference of the rate of modal chromosome number between mcs + n and mcs passages was not more than 5%-15% and the structure aberrations was generally 0%-3%, not more than 5%-10%, thus the hereditary character of cell lines is comparatively stable without significant difference between different passages. The genetic characteristics of chromosomal number of cell lines determines their tumorigenicity, but it is species specific. Experimental models were established for the researches on the prevention and prophylaxis of malignant tumors or cancers and their genetically biological characteristics. Tests showed that there was correlation among cell line chromosome number variations, anchorage independence in soft agar, agglutination under plant lectins and tumor-forming ability in nude mice. Since testing in vitro is more economic, simpler, faster, and is thought to be reliable, we recommend plant lectins followed by SA or analysis of karyotypes as the initial means for monitoring tumorigenicity of animal cell line in nude mice.

A 10 mm gap of rat sciatic nerve was created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes filled with an extracellular gel containing collagen, laminin and fibronectin. Empty silicone rubber tubes were used as controls. Six weeks after implantation, all extracellular elements were completely degraded and absorbed, and 90% of the animals from the extracellular gel group exhibited regeneration across the nerve gaps, whereas only 60% in the control group. Both qualitative and quantitative histology of the regenerated nerves revealed a more mature ultrastructural organization with 28% larger cross-sectional area and 28% higher number of myelinated axons in the extracellular gel group than the controls. These results showed that the gel mixture of collagen, laminin and fibronectin could offer a suitable growth medium for the regeneration of axons.

A sensitive technique for the sexing of bovine embryos was developed using polymerase chain reaction (PCR) amplification of the bovine amelogenin (bAML) gene on the X- and Y-chromosomes of Holstein dairy cattle. Cloning and DNA sequencing showed a 45.1% homology between the fifth intron of the bAML-X and bAML-Y gene with multiple deletions. A pair of sex-specific primers was designed to allow amplification of a single fragment of 467-bp from the X-chromosome of female cattle and two fragments of 467-bp and 341-bp from the X- and Y-chromosomes of male cattle. The primers were successfully applied to bovine sexing from single blastomeres isolated from day-6 to day-7 cow embryos by direct cell lysis and PCR. Our protocol of embryo sexing should be applicable to the diagnosis of defective genes in vitro in human embryos and in other domestic or recreational animals., (Copyright 1999 Wiley-Liss, Inc.)

This study investigated the carcase composition of Taiwan simulated native chickens at different stages of growth. The results indicated that the carcases of male chickens exhibited significantly larger protein mass and water mass than those of females during weeks 7 to 20; as was the ash mass during weeks 8 to 20. However, the fat mass of the carcases of female chickens was larger than that of males during weeks 14 to 20. Separate regression equations were derived for the relationship between protein mass, fat mass, ash mass and water mass and both carcase weight and age of both male and female chickens.

In the cerebral ischemic pathophysiologic mechanism, lactic acidosis is a important factor to exacerbate cerebral damage. Our research showed that the lactic level of cerebral cortex in rats increased rapidly after the focal cerebral ischemia or during blood reperfusion after cerebral ischemia, 26.99 +/- 5.89 and 28.63 +/- 5.08 mumol/g brain wight respectively, it exacerbated significantly brain edema and pathological damage. The lactic level decreased rapidly to treat with dichloroacetate (50 mg/kg body weight) after cerebral ischemia or during blood reperfusion, 14.11 +/- 2.06 and 13.23 +/- 1.71 mumol/g brain wight respectively, brain edema and pathology improved significantly. It suggested that dichloroacetate might across blood-brain barrier into the cerebral ischemic region and lowered lactic level, improved brain internal environment, relieved cerebral damage after focal cerebral ischemia or during blood reperfusion. It may improve the prognosis of patient with ischemic cerebral vascular disease to be treated with dichloroacetate early.

A technique is presented for feature extraction of a waveform y based on its Tauberian approximation, that is, on the approximation of y by a linear combination of appropriately delayed versions of a single basis function x, i.e., y(t) = ¿M i = 1 aix(t - ¿i), where the coefficients ai and the delays ¿i are adjustable parameters. Considerations in the choice or design of the basis function x are given. The parameters ai and ¿i, i=1, . . . , M, are retrieved by application of a suitably adapted version of Prony's method to the Fourier transform of the above approximation of y. A subset of the parameters ai and ¿i, i = 1, . . . , M, is used to construct the feature vector, the value of which can be used in a classification algorithm. Application of this technique to the classification of wide bandwidth radar return signatures is presented. Computer simulations proved successful and are also discussed.

The prevalence ratio, incidence and mortality rates from cerebrovascular disease (CVD) among 41,232 subjects, in rural areas of Sichuan, People's Republic of China, were investigated and various suspect risk factors to stroke were evaluated. The prevalence ratio of CVD was 266.10/100,000 population; annual incidence rate was 179.86/100,000 population; annual mortality rate was 153.02/100,000 population. The main risk factors were hypertension, heart diseases, sclerotic findings and TIAs. It was characterized by higher incidence, shorter course, higher mortality of stroke in rural area.

The endotracheal administration of 14C-proline to young adult golden hamsters was effective in radiolabeling collagen and resulted in a greater specific activity for hydroxyproline than did intraperitoneal administration of the amino acid. The isolation of purified isoluble collagen was accomplished with the aid of pancreatic elastase. Enzyme treatment of lung homogenates resulted in the solubilization of 35 per cent of total lung collagen. The residue obtained after elastase treatment was pure collagen. A significant portion of the radiolabeled collagen was solubilized by the elastase treatment, which suggests that elastase may be helpful in distinguishing between newly synthesized and total lung collagen. Studies performed at various times after administration of 14C-proline indicate that a greater portion of the collagen radioactivity is solubilized during the early days after treatment with the amino acid. The methodology and procedures outlined in this communication allow for additional insights into the metabolism of connective tissue, specifically collagen, in normal and diseased lungs.