Expression of hypoxia inducible factor-1 alpha and ischemic erythropoietin tolerance in the brain of cerebral ischemic tolerance model rats

BACKGROUND： Hypoxia inducible factor-1 alpha （HIF-1 （x） and erythropoietin（EPO）, possessing neuroprotective effect in the cerebral ischemia, might play an important role in the formation of cerebral ischemic tolerance （IT）. OBJECTIVE：To observe the neuroprotective effect of cerebral ischemic preconditioning（IPC） of rats, and the expression and mechanism of HIF-1α and target gene erythropoietin in the brain tissue following the formation of cerebral IT. DESIGN ： A randomized and controlled observation SETTING： Department of Neurology, the Affiliated Hospital of Medical College, Qingdao University MATERIALS： Totally 84 enrolled adult healthy male Wistar rats of clean grade, weighing 250 to 300 g, were provided by the Animal Experimental Department, Tongji Medical College of Huazhong University of Science and Technology. Ready-to-use SABC reagent kit and rabbit anti-rat HIF-1α monoclonal antibody were purchased from Boshide Bioengineering Co.Ltd （Wuhan）; Rabbit anti-rat EPO monoclonal antibody was purchased from Santa Cruz Company （USA）. METHODS： This experiment was carried out in the Department of Anatomy, Medical College, Qingdao University during March 2005 to March 2006. ① The 84 rats were divided into 3 groups by a lot： IPC group （n=40）, sham-operation group （n=40） and control group （n=4）. In the IPC group, middle cerebral artery was occluded for 2 hours respectively on the 1^st, 3^rd, 7^th, 14^th and 21^st days of the reperfusion following 10-minute preischemia was made using a modified middle cerebral artery second suture method from Zea-Longa. The rats were sacrificed 22 hours after reperfusion in the end of middle cerebral artery occlusion （MCAO）. That was to say, after 10-minute preischemia, suture was exited to the extemal carotid artery and embedded subcutaneously. Middle cerebral artery was occluded again to form the second reperfusion at the set time point after reperfusion. Twenty-two hours later, rats were sacrificed; In the sham-operation group,the preischemia was substituted by sham-operation（only common carotid artery and crotch were exposed, and MCAO by suture was omitted）, and the other procedures were the same as those in the IPC group. In the control group, rats were given sham-operation twice at an interval of one day, and they were sacrificed 24 hours after the second sham-operation. ② Brain tissue was taken from the rats in each group. Cerebral infarction area of each layer was measured with TTC staining, and total cerebral infarction volume （The total cerebral infarction area of each layerxinterspace ） was calculated. After brain tissue was stained by haematoxylin-esoin （HE）, the form of nerve cells was observed under an optical microscope, and the expressions of HIF-1α（and EPO protein in the brain tissue were detected with immunohistochemical method. MAIN OUTCOME MEASURES： ①Cerebral infarction volume;②form of nerve cell; ③ the expression of HIF-1α and EPO protein in the brain tissue. RESULTS：Totally 84 rats were enrolled in the experiment. The dead rats were randomly supplied during the experiment, and finally 84 rats entered the stage of result analysis. ① Detection of cerebral infarction volume of rats in each group： Cerebral infarction volume in the IPC group was significantly smaller than that in the sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively [（161.2±6.9） mm^3 vs （219.9±11.2） mm^3, （134.9±9.0） mm^3 vs （218.6±13.0） mm^3, （142.9±13.7） mm^3 vs （221.3±14.2） mm^3, t=-8.924, 10.587,7.947, P〈 0.01]. ② Observation of nerve cell form of brain tissue： HE staining showed that the ischemic degree, range and cerebral edema degree of IPC group were significantly milder than those of sham-operation group. ③ The expressions of HIF-1α and EPO protein in cerebral cortex and hippocampus ： The expression of HIF-1αof IPC group was significantly higher than that of sham-operation group on the 1^st, 3^rd and 7^th days after reperfusion respectively （125.93±3.79 vs 117.65±5.60, 140.63±4.64 vs 119.33±4.26, 131.15±2.74 vs 107.60±3.89, t=2.449, 6.763,9.899,P 〈 0.05-0.01）. The expression of EPO of IPC group was significantly higher than that of sham-operation group on the 3^rd and 7^th days after perfusion respectively （141.68±3.29 vs 126.33±4.51, 138.88±2.59 vs 125.58±6.18,t=5.499,3.970, P〈 0.05）. CONCLUSION ： ①IPC can protect the never cells in rat brain and the best time to onset of cerebral IT induced by IPC is 1 to 7 days after reperfusion. ② Neuroprotective effect of cerebral IT might be related to the expression of HIF-1α and its target gene EPO.

Deterioration in Hemodynamics Reaction, Baroreflex Sensitivity, Sympathetic Nerve Activity and Redox State of Thoracic Aorta in the Experimental Model of Nitrate Tolerance and Its Pharmacological Correction

Continuous treatment with organic nitrates causes nitrate tolerance and provides evidence for a relationship between mitochondrial complex 1 activity and mitochondrial aldehyde dehydrogenase-2 (ALDH-2) with disturbances of the hemodynamics reaction during nitroglycerin (NTG) tolerance (NTGT). The purpose of this study was the evaluation of efficacy of original oxidized form NAD-containing drug, NADCIN®, on hemodynamic reactions, baroreflex sensitivity (BRS) and reflex control of splanchnic sympathetic nerve activity (SSNA), level of redox-potential, activity of ALDH-2 and superoxide anion generation in aortic tissue in rat model of NTGT. Five groups (7 - 9 each) of male Wistar rats, including control, acute i.v. NTG (150 mcg/kg) administration, NTG tolerance NTGT treatment with NADCIN® 8 mg/kg and methylene blue (MB, 2.5 mg/kg) were used. NTGT in rats was accompanied with the greatly attenuation of hemodynamics reaction, BRS, the decreasing of the ability to reflex control of SSNA without pronounce overexpression of endothelin-1 in vessels (aorta). In NTGT rats i.v. NTG along induced less hypotensive reactions and alterations in heart period vs single NTG treated group, more expressively decreased BRS (-34%) and reflex control of SSNA (-18%). NADCIN® significantly inhibits tolerance-inducing properties of the prolonged nitroglycerin infusion (max decrease of blood pressure response to nitroglycerin injection, % of normal controls: NTGT 51.2%, NADCIN® 91.6%, MB 55.8%). NADCIN® in NTGT rats after NTG i.v. administration increased reduced BRS (+37.8%, p < 0,05), reflex control of SSNA (+29.4%, p < 0.05) and reversed the decreasing of NAD/NADH ratio, ALDH-2 activity and decreasing in superoxide generation in thoracic aortic tissue. Thus, course treatment with NADCIN® of NTGT rats restores hemodynamics changes, BRS and SSNA throughout the increasing of redox-potential NAD/NADH and cessates the NTGT developing.

Objective function of cost in optimal tolerance allocation

An objective function model is proposed for cost in optimizing and allocating tolerance with consideration of manufacturing conditions. With the fuzzy comprehensive evaluation method,a manufacturing difficulty coefficient is derived,which takes into account of several factors affecting the manufacturing cost,including the forming means of the blank,size,machining surface features,operator’s skills and machinability of materials. The coefficient is then converted into a weight factor used in the inversed square model representing the relationship between the cost and tolerance,and,hence,an objective function for cost is established in optimizing and allocating tolerance. The higher is the manufacturing difficulty coefficient,the higher is the relative manufacturing cost and the higher is the weight factor of the tolerance allocation,which indicates the increase of the tolerance’s effects on the total manufacturing cost and,therefore,a larger tolerance should be allocated. The computer-aided tolerance allocation utilizing this model makes it more convenient,accurate and practicable.

Physical layer impairments tolerance based lightpath provision in software defined optical network

As all-optical networks grow with ever increasing ultra-high speed,the communication quality suffers seriously from physical layer impairments( PLIs). The same problem still exists in software defined optical network( SDON) controlled by OpenFlow. Aimed to solve this problem,a PLIs tolerance based lightpath provision scheme is proposed for OpenFlow controlled optical networks. This proposed approach not only takes the OSNR model to represent those linear PLIs factors,but also introduces those nonlinear factors into the OSNR model. Thus,the proposed scheme is able to cover most PLIs factors of each optical link and conduct optical lightpath provison with better communication quality. Moreover,PLIs tolerance model is also set up and considered in this work with some necessary extension to OpenFlow protocols to achieve better compatibility between physical layer impairments factors and various services connections. Simulation results show that the proposed scheme is able to get better performance in terms of packet loss rate and connection setup time.

Accuracy Analysis and Design of A3 Parallel Spindle Head

As functional components of machine tools, parallel mechanisms are widely used in high efficiency machining of aviation components, and accuracy is one of the critical technical indexes. Lots of researchers have focused on the accuracy problem of parallel mechanisms, but in terms of controlling the errors and improving the accuracy in the stage of design and manufacturing, further efforts are required. Aiming at the accuracy design of a 3-DOF parallel spindle head（A3 head）, its error model, sensitivity analysis and tolerance allocation are investigated. Based on the inverse kinematic analysis, the error model of A3 head is established by using the first-order perturbation theory and vector chain method. According to the mapping property of motion and constraint Jacobian matrix, the compensatable and uncompensatable error sources which affect the accuracy in the end-effector are separated. Furthermore, sensitivity analysis is performed on the uncompensatable error sources. The sensitivity probabilistic model is established and the global sensitivity index is proposed to analyze the influence of the uncompensatable error sources on the accuracy in the end-effector of the mechanism. The results show that orientation error sources have bigger effect on the accuracy in the end-effector. Based upon the sensitivity analysis results, the tolerance design is converted into the issue of nonlinearly constrained optimization with the manufacturing cost minimum being the optimization objective. By utilizing the genetic algorithm, the allocation of the tolerances on each component is finally determined. According to the tolerance allocation results, the tolerance ranges of ten kinds of geometric error sources are obtained. These research achievements can provide fundamental guidelines for component manufacturing and assembly of this kind of parallel mechanisms.

Quantitative Analysis of Multi-Recovery-Based Intrusion Tolerance Model

Quantitative analysis has always been a difficult problem in security analysis of intrusion tolerance systems. An intrusion tolerance model based on multiple recovery mechanisms is introduced in this paper and how to quantify the security attributes of the model is proposed. A state transition model with recovery states more accurately describes the dynamic behavior of the system. Considering that recovery mechanisms have a great impact on the security performance of the system, we set up the cost models corresponding to different recovery mechanisms. We propose a feasible security measure based on mean cost to security failure in order to evaluate the system cost during the recovery phase. The experimental results confirmed the feasibility of the proposed methods.

Intracellular detection of cytokine expression with four-color flow cytometry in pregnant NOD/SCID mice

The aim of this study was to evaluate the relationship between the embryo resorption rate （RR） and the status of local immunity at the feto-maternal interface in pregnant NOD/SCID mice. RR was calculated in NOD/SCID x NOD/SCID mice and compared with non-immunodeficiency BALB/c × BALB/c mice on day 13.5 of gestation. Intraeellular detection of cytokine expression was performed with four-color flow cytometry to identify the functional subsets of lymphocytes in NOD/SCID mice before being bred or during pregnancy. No statistically supported difference in RRs was observed between NOD/SCID × NOD/SCID and control BALB/c × BALB/c mice. Accordingly, although multiple immunodeficits were confirmed in NOD/SCID mice, the percentages of several functional cell subsets were spontaneously altered during pregnancy, and this may be correlated with the roughly normal pregnancy outcomes observed in these mice. The spontaneous alteration of cell percentages at the feto-maternal interfaee in NOD/SCID mice may be of benefit to the pregnancy outcomes.