Transcranial Doppler Ultrasonography detection on cerebrovascular flow for evaluating neonatal hypoxic-ischemic encephalopathy modeling.

Objective: This study aimed to investigate the feasibility of Transcranial Doppler Ultrasonography (TCD) in evaluating neonatal hypoxic-ischemic encephalopathy (NHIE) modeling through monitoring the alteration of cerebrovascular flow in neonatal hypoxic-ischemic (HI) rats. Methods: Postnatal 7-day-old Sprague Dawley (SD) rats were divided into the control group, HI group, and hypoxia (H) group. TCD was applied to assess the changes of cerebral blood vessels, cerebrovascular flow velocity, and heart rate (HR) in sagittal and coronal sections at 1, 2, 3, and 7 days after the operation. For accuracy, cerebral infarct of rats was examined by 2,3,5-Triphenyl tetrazolium chloride (TTC) staining and Nissl staining to simultaneously verify the establishment of NHIE modeling. Results: Coronal and sagittal TCD scans revealed obvious alteration of cerebrovascular flow in main cerebral vessels. Obvious cerebrovascular back-flow was observed in anterior cerebral artery (ACA), basilar artery (BA), middle cerebral artery (MCA) of HI rats, along with accelerated cerebrovascular flows in the left internal carotid artery (ICA-L) and BA, decreased flows in right internal carotid artery (ICA-R) relative to those in the H and control groups. The alterations of cerebral blood flows in neonatal HI rats indicated successful ligation of right common carotid artery. Besides, TTC staining further validated the cerebral infarct was indeed caused due to ligation-induced insufficient blood supply. Damage to nervous tissues was also revealed by Nissl staining. Conclusion: Cerebral blood flow assessment by TCD in neonatal HI rats contributed to cerebrovascular abnormalities observed in a real-time and non-invasive way. The present study elicits the potentials to utilize TCD as an effective means for monitoring the progression of injury as well as NHIE modeling. The abnormal appearance of cerebral blood flow is also beneficial to the early warning and effective detection in clinical practice.

Research progress of the CXCR4 mechanism in Alzheimer's disease.

Alzheimer's disease (AD) is a degenerative brain disease with complex clinical manifestations and pathogeneses such as abnormal deposition of beta-amyloid protein and inflammation caused by the excessive activation of microglia. CXC motif chemokine receptor type 4 (CXCR4) is a type of G protein-coupled receptor that binds to CXC motif ligand 12 (CXCL12) to activate downstream signaling pathways, such as the Janus kinase/signal transducer and activator of transcription and the renin-angiotensin system (Ras)/RAF proto-oncogene serine (Raf)/mitogen-activated protein kinase/extracellular-regulated protein kinase; most of these signaling pathways are involved in inflammatory responses. CXCR4 is highly expressed in the microglia and astrocytes; this might be one of the important causes of inflammation caused by microglia and astrocytes. In this review, we summarize the mechanism and therapeutics of AD, the structures of CXCR4 and the CXCL12 ligand, and the mechanisms of CXCR4/CXCL12 that are involved in the occurrence and development of AD. The possible treatment of AD through microglia and astrocytes is also discussed, with the aim of providing a new method for the treatment of AD.

Bioinformatics analysis of the wheel treadmill test on motor function recovery after spinal cord injury.

This study aimed to explore the possible target and mechanism of the wheel treadmill (WTM) test for motor function recovery of spinal cord injury (SCI). Rats were divided into sham, control and WTM groups to establish an SCI mode. Rats in the WTM group were trained on the WTM test, and Basso-Beattie-Bresnahan (BBB) scores were determined. The samples were collected, and mRNA sequencing was conducted to determine the changes in gene expression. The coexpressed genes were screened to construct a protein-protein interaction (PPI), followed by the Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology function enrichment analysis, and the differentially expressed genes (DEGs) volcano map and hub gene expression heat map were constructed using R language. The BBB scores in the control and WTM groups increased with time, with the WTM group scoring higher than the control group. The results of rat spinal cord tissue sequencing showed that a total of 1679 DEGs were screened in the sham and control groups; 928 DEGs and 731 overlapping genes were screened in the WTM and control groups. The key genes were identified by PPI analysis. One hundred and thirty-three genes were found to be overlapping by combined analysis of spinal cord sequencing data and BBB scores of rats at Week 7. The top 10 DEGs from high to low were Tyrobp, Rac2, Cd68, C1qb, Aif1, Cd74, Spi1, Fcer1g, RT1-DA, and Ccl4. The terms with the highest enrichment scores were microglia-mediated positive regulation of cytotoxicity and major histocompatibility complex class II protein complexes. Treatment with the WTM test promotes recovery of motor function after SCI in rats by modulating intercellular communication and immune function.

Catalytic decomposition of gaseous 1,2-dichlorobenzene over CuOx/TiO2 and CuOx/TiO2-CNTs catalysts: Mechanism and PCDD/Fs formation.

Gaseous 1,2-dichlorobenzene (1,2-DCBz) was catalytically decomposed in a fixed-bed catalytic reactor using composite copper-based titanium oxide (CuOx/TiO2) catalysts with different copper ratios. Carbon nanotubes (CNTs) were introduced to produce novel CuOx/TiO2-CNTs catalysts by the sol-gel method. The catalytic performances of CuOx/TiO2 and CuOx/TiO2-CNTs on 1,2-DCBz oxidative destruction under different temperatures (150-350 °C) were experimentally examined and the correlation between catalyst structure and catalytic activity was characterized and the role of oxygen in catalytic reaction was discussed. Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) generation during 1,2-DCBz catalytic oxidation by CuOx/TiO2-CNTs composite catalyst was also examined. Results indicate that the 1,2-DCBz destruction/removal efficiencies of CuOx (4 wt%)/TiO2 catalyst at 150 °C and 350 °C with a GHSV of 3400 h(-1) are 59% and 94% respectively and low-temperature (150 °C) catalytic activity of CuOx/TiO2 on 1,2-DCBz oxidation can be improved from 59 to 77% when CNTs are introduced. Furthermore, oxygen either in catalyst or from reaction atmosphere is indispensible in reaction. The former is offered to activate and oxidize the 1,2-DCBz adsorbed on catalyst, thus can be generally consumed during reaction and the oxygen content in catalyst is observed lost from 39.9 to 35.0 wt% after reacting under inert atmosphere; the latter may replenish the vacancy in catalyst created by the consumed oxygen thus extends the catalyst life and raises the destruction/removal efficiency. The introduction of CNTs also increases the Cu(2+)/Cu(+) ratio, chemisorbed oxygen concentration and surface lattice oxygen binding energy which are closely related with catalytic activity. PCDD/Fs is confirmed to be formed when 1,2-DCBz catalytically oxidized by CuOx/TiO2-CNTs composite catalyst with sufficient oxygen (21%), proper temperature (350 °C) and high concentration of 1,2-DCBz feed (120 ppm).

Lacidipine attenuates TNF-α-induced cardiomyocyte apoptosis.

This study was designed to investigate whether lacidipine elicited a protective role on cardiomyocyte against apoptosis induced by TNF-α. Neonatal rat cardiomyocytes were randomly assigned into different groups. TUNEL staining was utilized to detect apoptosis, and caspase-3 and caspse-12 were determined. To explore the underlying mechanism, Z-ATAD-FMK (a selective caspase-12 inhibitor) was used to identify the key molecule involved. TNF-α increased caspase-3 expression, which was mediated by increased caspase-12 expression. In the meantime, apoptosis was significantly induced by TNF-α. Lacidipine lowered caspase-12 and caspase-3 expression, and cardiomyocyte apoptosis induced by TNF-α. The results suggest that lacidipine attenuates TNF-α -induced apoptosis via inhibition of caspase-12 and caspase-3 successively.

Anti-arrhythmic effect of verapamil is accompanied by preservation of cx43 protein in rat heart.

The present study was to test the hypothesis that anti-arrhythmic properties of verapamil may be accompanied by preserving connexin43 (Cx43) protein via calcium influx inhibition. In an in vivo study, myocardial ischemic arrhythmia was induced by occlusion of the left anterior descending (LAD) coronary artery for 45 min in Sprague-Dawley rats. Verapamil, a calcium channel antagonist, was injected i.v. into a femoral vein prior to ischemia. Effects of verapamil on arrhythmias induced by Bay K8644 (a calcium channel agonist) were also determined. In an ex vivo study, the isolated heart underwent an initial 10 min of baseline normal perfusion and was subjected to high calcium perfusion in the absence or presence of verapamil. Cardiac arrhythmia was measured by electrocardiogram (ECG) and Cx43 protein was determined by immunohistochemistry and western blotting. Administration of verapamil prior to myocardial ischemia significantly reduced the incidence of ventricular arrhythmias and total arrhythmia scores, with the reductions in heat rate, mean arterial pressure and left ventricular systolic pressure. Verapamil also inhibited arrhythmias induced by Bay K8644 and high calcium perfusion. Effect of verapamil on ischemic arrhythmia scores was abolished by heptanol, a Cx43 protein uncoupler and Gap 26, a Cx43 channels inhibitor. Immunohistochemistry data showed that ischemia-induced redistribution and reduced immunostaining of Cx43 were prevented by verapamil. In addition, diminished expression of Cx43 protein determined by western blotting was observed following myocardial ischemia in vivo or following high calcium perfusion ex vivo and was preserved after verapamil administration. Our data suggest that verapamil may confer an anti-arrhythmic effect via calcium influx inhibition, inhibition of oxygen consumption and accompanied by preservation of Cx43 protein.

Effect of pomegranate peel polyphenol gel on cutaneous wound healing in alloxan-induced diabetic rats.

BACKGROUND: Pomegranate (punica granatum) belongs to the family Punicaceae, and its peel has been used as a traditional Chinese medicine because of its efficacy in restraining intestine, promoting hemostasis, and killing parasites. Pomegranate peel has been reported to possess wound-healing properties which are mainly attributed to its polyphenol extracts. The purpose of this study was to investigate the effect of pomegranate peel polyphenols (PPP) gel on cutaneous wound healing in diabetic rats. METHODS: Alloxan-induced diabetic rats were given incisional wounds on each side of the mid-back and then treated daily with PPP gel (polyphenol mass fraction = 30%) post-wounding. Rats were sacrificed on days 4, 7, 14, and 21 post-wounding to assess the rates of wound closure, histological characteristics; and to detect the contents of hydroxyproline, production of nitric oxide (NO), and activities of NO synthase (NOS), as well as the expressions of transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) in wound tissue. RESULTS: Wound closure was significantly shortened when PPP gel was applied to the wounds of diabetic rats. Histological examination showed the ability of PPP gel to increase fibroblast infiltration, collagen regeneration, vascularization, and epithelialization in the wound area of diabetic rats. In addition, PPP gel-treated diabetic rats showed increased contents of hydroxyproline, production of NO, and activities of NOS and increased expressions of TGF-ß1, VEGF, and EGF in wound tissues. CONCLUSION: PPP gel may be a beneficial method for treating wound disorders associated with diabetes.

Κ-opioid receptor stimulation improves endothelial function in hypoxic pulmonary hypertension.

The present study was designed to investigate the effect of κ-opioid receptor stimulation with U50,488H on endothelial function and underlying mechanism in rats with hypoxic pulmonary hypertension (HPH). Chronic hypoxia-induced HPH was simulated by exposing the rats to 10% oxygen for 2 wk. After hypoxia, mean pulmonary arterial pressure (mPAP), right ventricular pressure (RVP) and right ventricular hypertrophy index (RVHI) were measured. Relaxation of pulmonary artery in response to acetylcholine (ACh) was determined. Expression and activity of endothelial nitric oxide (NO) synthase (eNOS) and inducible NO synthase (iNOS) with NO production, total antioxidant capacity (T-AOC), gp91(phox) expression and nitrotyrosine content were measured. The effect of U50,488H administration during chronic hypoxia was investigated. Administration of U50,488H significantly decreased mPAP and right ventricular hypertrophy as evidenced by reduction in RVP and RVHI. These effects were mediated by κ-opioid receptor. In the meantime, treatment with U50,488H significantly improved endothelial function as evidenced by enhanced relaxation in response to ACh. Moreover, U50,488H resulted in a significant increase in eNOS phosphorylation, NO content in serum, and T-AOC in pulmonary artery of HPH rats. In addition, the activity of eNOS was enhanced, but the activity of iNOS was attenuated in the pulmonary artery of chronic hypoxic rats treated with U50,488H. On the other hand, U50,488H markedly blunted HPH-induced elevation of gp91(phox) expression and nitrotyrosine content in pulmonary artery, and these effects were blocked by nor-BNI, a selective κ-opioid receptor antagonist. These data suggest that κ-opioid receptor stimulation with U50,488H improves endothelial function in rats with HPH. The mechanism of action might be attributed to the preservation of eNOS activity, enhancement of eNOS phosphorylation, downregulation of iNOS activity and its antioxidative/nitrative effect.

Photocatalytic decomposition on nano-TiO2: destruction of chloroaromatic compounds.

Photocatalysis is applied increasingly in addressing and solving environmental and energy-related problems. Especially the TiO2-derived catalysts attract attention because of their catalytic efficiency, wide range of applications, ease in use, and low cost (it costs about 150 Yuan a kilogram in China). This review first describes the principles of photocatalytic destruction by semiconductors and then focuses on degradation rates and reaction mechanisms in a variety of photocatalytic uses of modified TiO(2). Finally, these concepts are illustrated by selected examples relating to the photocatalytic degradation of organic persistent pollutants, such as polychlorinated benzenes (PCBz), biphenyls (PCB) and dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). And some approaches towards industrial application are analyzed.

[Experimental study on the preventive and treatment function of cortex mori for peripheral nervous lesion at the early stage of diabetes rats].

AIM: To observe the preventive function of cortex mori for peripheral nervous lesion at the early stage in diabetes rats, and probe into the mechanism of that formula. METHODS: Set up the diabetes rat model evoked by alloxan. According to different blood sugar values, randomly divide the rats into normal group, model group, cortex mori(high dosage and low dosage) group and methylcobalamin control group, respectively administer the rats with saline and cortex mori of different dosages by ig (1.875 g/kg, 0.625 g/kg), while 300 microg/kg methylcobalamin for control group, one time each day. Two months after the administration, determine the FBG, body weight, SOD and MDA in blood serum of rats in each group, observe the changes on final product of glycosylation, CGMP and CAMP of sciatic nerve and the synapsin of rats' sciatic nerves. Conduct the pathological observation on area of myelin sheath, extramedullary fiber and the cross section of myelin sheath of sciatic nerves. And observe the changes of ultrafine form of sciatic nerve through transmission electron microscope. In the mean time, determine the MNCV, SNVC and SL, and the tail-flicking test should be undertaken for checking the sensory nerve. RESULTS: Cortex mori can effectively enlarge the area of myelin sheath, extramedullary fiber and the cross section of myelin sheath. CONCLUSION: Cortex mori can obviously ease up the pathological changes of peripheral nerve at the early stage of the diabetes rats, and the overall curative effect is better than that of methylcobalamin.

[Plasma glutathione of patients with coronary heart disease measured by fluorospectrophotometer].

The standard samples of reduction form glutathione (GSH) and oxidization form glutathione (GSSG) were scanned with full-wavelength range to determine the excitation wavelength lambda(ex) 334.4 nm, the emission wavelength lambda(em) 424 nm, and the spectral bandwidth 5 nm respectively. Phosphate buffer saline (PBS) of pH 8. 3 served as buffer solution. GSH was incubated for 30 min with 100 microL o-phthaldehyde (OPA) of 10 mmol x L(-1) methyl alcohol solution for derivatization, and then fluorescence intensities were measured. With standard glutathione concentration being independent variable and fluorescence intensity being dependent variable, the linear equations for GSH and GSSG were deduced: Y(GSH) = 6.9 + 8.6X (r2 = 0.994) and Y(GSSG) = 6.2 + 17.2X (r2 = 0.999). Standard curves were done hereby. The plasma glutathione of three groups was then measured, and GSH/ GSSG redox potential was calculated according to Nernst equation. The results indicated that, from normal control group to prophase coronary heart disease group, then to coronary heart disease group, the GSH and GSH/GSSG ratio gradually reduced, GSSG and GSH/GSSG redox potential gradually increased (more positive) (all P < 0.05), and the redox potential shifted to oxidization direction along with the development of coronary heart disease. This fluorospectrophotometry method showed simple operation, and fine precision and accuracy.

Effect of ozone produced from antibody-catalyzed water oxidation on pathogenesis of atherosclerosis.

Recent studies have suggested that antibodies can catalyze the generation of unknown oxidants including hydrogen peroxide (H2O2) and ozone (O3) from singlet oxygen (1O2) and water. This study is aimed to detect the effect of antibody-catalyzed water oxidation on atherosclerosis. Our results showed that both H2O2 and O3 were produced in human leukemia THP-1 monocytes incubated with human immunoglobulin G and phorbol myristate acetate. In the THP-1 monocytes incubated with human immunoglobulin G, phorbol myristate acetate and low density lipoprotein, the intracellular total cholesterol, free cholesterol, cholesteryl ester and lipid peroxides clearly increased, and a larger number of foam cells were observed by oil red O staining. The accumulation of all intracellular lipids was significantly inhibited by vinylbenzoic acid, and only slightly affected by catalase. These findings suggested that the production of O3, rather than H2O2, might be involved in the pathogenesis of atherosclerosis through the antibody-catalyzed water oxidation pathway.

[Establishment of rat model of psychical erectile dysfunction].

OBJECTIVE: To set up a method of establishing the animal model of psychical erectile dysfunction with emotional stress. METHODS: All thirty-six male rats with normal sexual function were divided into three groups, i. e. normal group, model group and demasculinized group randomly according to their weights. The rats in the model group were suspended upside down in midair over the water and irritated repeatedly. Two weeks later, the sexual abilities of all rats, i. e. the times of mounting and intromitting the estrus female rats, the latent period of mounting, intromission and ejaculation, were recorded, and the number of rats that had sexual activities was also counted. And the hemorheology indices of the rats were measured. RESULTS: Compared with the normal rats, the latency of mounting [(152.5 +/- 24.6) s vs (42.4 +/- 9.6) s] and intromission [(437.0 +/- 67.7) s vs (130.8 +/- 39.1) s] of the model rats were longer (P < 0.01), but the latency of ejaculation [(385.3 +/- 80.0) s vs (547.3 +/- 69.4) s] was shorter (P < 0.05) than that of the normal. There was no significant difference in the times of mounting between normal [(38.3 +/- 6.1) vices and model rats (38.5 +/- 5.4) vices], but the intromission times of model rats [(9.2 +/- 1.7) vices] was lower than that of the normal rats [(20.3 +/- 3.1) vices], P < 0.01. Compared with the normal rats, the sexual activity incidence of the model rats (mounting: 58.3%, intromission: 33.3%, ejaculation: 16.7%) was significant lower than that of the normal rats (100%) (P < 0.01). But there was no significant difference in the sexual ability between the model and the demasculinized rats (P > 0.05). The hemorheology indices, e. g. blood viscosity, hematocrit (Hct) and red cell aggregation (RCA), of the model rats was significant higher than that of the normal and demasculinized rats (P < 0.05), but there was no significant difference between the normal and demasculinized rats. CONCLUSION: The rat model of psychical erectile dysfunction can be made ideally with psychical stress.

[Antibody-induced ozone production and its potential effects on pathogenesis of atherosclerosis].

In the present study, we measured the antibody-catalyzed 03 formation from THP-1 monocytes activated by phorbol myristate acetate (PMA) by indigo carmine bleaching reaction test, and the accumulation of cholesterol in THP-1 monocytes by fluorescence spectrophotometric method, and analyzed the cholesterol ozonation product 5,6-secosterol by high-performance liquid chromatography (HPLC), to explore the potential effect of antibody-catalyzed water oxidation on pathogenesis of atherosclerosis. It was showed that THP-1 monocytes incubated with human IgG and PMA evidently produced an oxidant with the chemical signature of 03 which could bleach indigo carmine, and be intensified or inhibited respectively by catalase and vinylbenzoic acid. In the THP-1 monocytes incubated with human IgG, PMA and LDL, the intracellular accumulated total cholesterol (TC), free cholesterol (FC), cholesteryl ester (CE) and the CE/TC increased evidently, and the cholesterol ozonation product 5,6-secosterol was also produced markly, all of that were inhibited by vinylbenzoic acid. These results demonstrated that the activated THP-1 monocytes possess the ability to produce O3 through antibody-catalyzed water-oxidation pathway, which could be a new mechanism concerned with atheriosclerosis.

[Effect of triptolide on expression of urokinase-type plasminogen activators in vascular endothelial cells].

BACKGROUND & OBJECTIVE: It has been showed that triptolide (T10) has antitumor activities. This study was to observe the inhibitory effects of T10 on proliferation of vascular endothelial cells and expression of urokinase-type plasminogen activator (u-PA), and to elucidate the antiangiogenic mechanism of T10. METHODS: Human umbilical vein endothelial cells were cultured in vitro for 3 generations, and treated with T10 (0, 5, 10, 20, and 30 microg/L), or dexamethasone (300 mg/L) as control. Cell count and 3H-TDR incorporation were used to observe cell proliferation. The chick embryo chorioallantoic membrane (CAM) test was carried out to observe the effect of T10 on angiogenesis. Immunohistochemistry and real-time quantitive reverse transcription-polymerase chain reaction (RT-PCR) were used to detect protein and mRNA levels of u-PA. RESULTS: T10 inhibited the proliferation of umbilical vein endothelial cells in a dose-dependent manner; inhibitory rate of endothelial cells was 28.93% after treatment of 5 microg/L of T10. T10 significantly reduced angiogenesis in CAM, even at the concentration of 5 microg/L (P<0.05). T10 decreased protein and mRNA levels of u-PA in endothelial cells in a dose-dependent manner; the protein level of u-PA was reduced by 41.05% after treatment of 5 microg/L of T10. CONCLUSION: T10 could inhibit the proliferation of human umbilical vein endothelial cells and expression of u-PA, which may contribute to its antiangiogenic activity.

[Effects of triptolide on migration of vascular endothelial cells].

OBJECTIVE: To investigate the inhibitory effects of triptolide on the migration, gene and protein expression of urokinase-type plasminogen activator (u-PA) of endothelial cells (ECs) cultured in vitro and to elucidate the anti-angiogenic mechanism of triptolide. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and the passage 3 cells were treated with triptolide (0, 5, 10, 20, 30 microg/L). Three-dimensional culture system was used to assess the effect of triptolide on the migration of HUVECs. Real time quantitive reverse transcription polymerase chain reaction and immunohistochemical assay were applied to detect gene and protein expression of u-PA. RESULTS: Triptolide decreased the migration activity and suppressed the expression of u-PA of HUVECs at the gene and protein level. CONCLUSION: Triptolide could inhibit the migration of ECs by reducing the gene and protein expression of u-PA. These may contribute to an elucidation of the mechanism by which triptolide inhibits the angiogenesis.

[A novel fluorophotometric method for assaying free glutathione of both reduced and oxidized in plasma].

To investigate the measurement of free glutathione of both reduced or oxidized (i.e. GSH and GSSG) in plasma, and to evaluate the redox state of GGSH/GSSG in human plasma, both GSH and GSSG in plasma were measured by fluorophotometry based on the facts that one molar GSSG can be reduced to two molar GSH by dithiothreitol(DTT) under the condition of the pH being about 6.0, and GSH can provide both primary amine and thiol groups to react with the two carbonyl groups of O-phthaldehyde (OPA) to form a fluorescent ternary isoindole complex at pH 8.0. This method can at least measure 16 picomole GSH and 8 picomole GSSG respectively in the tube. The variation coefficient (CV) for intra-ssay and intera-ssay is about 4. 6% and 3.9% for GSH and 3.5% and 4.1% for GSSG respectively. The recovery of GSH and GSSG added to the plasma is (99.77% +/- 5.70)% and (99.28% +/- 4.73)% respectively. The concentration of GSH and GSSG in the plasma of young healthy volunteer is (16.5 +/- 2.4) nmol x mL(-1) and (1.7 +/- 0.35)nmol x mL(-1) respectively, without significant difference between male and female. This measurement method is simple with great sensitivity and selectivity for rapid measuring GSH and GSSG in human plasma simultaneously.