Study On The Relationship Between Individual Difference And The Biomarkers For Stress Response

Stress has influence on health, behavior and working efficiency, and it has become one of important international occupational health problem. So, the evaluation of stress level has significance meaning. Literature research indicated that, as a kind of personality traits, emotional stability has obvious individual differences and with relatively stable for each individual. This kind of personality stability in cognitive appraisal, personality, temperament type has a physiological basis. With the development of the modern life science, the understanding of stress research, from observation and description of phenomenon, has gradually developed to multidisciplinary angles which relate to "psychological-neuroendocrine-behavior", and reveal the response characteristics of stress and the deep mechanism of psychological and behavioral activities.The following five aspects of research were studied in this paper:(1) Development or optimization of technique platform for stress research; (2) Effects of exercise on restrained stress-induced changes of the brain monoamine neurotransmitters and behaviors in mice; (3) Effectof exercise stress on human stress bimarker response;(4) Multiple parameters for evaluation of human mental stress; (5) Salivary alpha-amylase, heart-rate variability and behavior reaction in acute mild psychosocial stress and the impact of Tridimensional Personality; (6) Stress reduced the capacity of attention responses regulation. This research projects will provide knowledge and technical supports for physiology, psychiatry, sports science and psychology researches. The main aspects of this paper were summarized as follows.1. Development or optimized of technique platform for stress research(1) Development or optimized rapid and automated analysis method of stress biomarker: ① Development of nanobody-based flow injection chemiluminescence immunoassay for sensitive detection of human prealbumin. Nanobodies, derived from camelid heavy-chain antibodies, have novel and impactful applications in clinical diagnostics. Our objective is to develop a nanobody-based chemiluminescence immunoassay for sensitive detection of human prealbumin (PA). In this context, a phage display nanobody library is constructed via immunizing dromedary camel with human prealbumin. Three nanobodies have been identified by five successive bio-panning steps. Based on their high expression level and good affinity, two out of three are chosen for further study. Magnetic beads (MBs) were functionalized with PEI by acylamide bond formed between the carboxyl groups on the surface of the MB. Then, the anti-PA nanobody (Nb1) can be effectively immobilized onto the surface of the functionalized MB using glutaradehyde as the linkage. The modified MBs with Nb1 can specifically capture the target PA and reacted with silica nanoparticles with co-immobilized HRP and anti-PA nanobody (Nb2). The concentration of PA was detected by flow injection chemiluminescence. When using MB/PEI as the carrier of anti-PA Nb1, the CL signal significantly increased to 4-fold compared with the signal using MB without PEI modification. The CL signal was further amplified to 5-fold when SiO2/Nb2 was used as the signal probe. Under the optimized conditions, the present immunoassay exhibited a wide quantitative range from 0.05 to 1000 μg L-1 with a detection limit of 0.01 μg L-1. The sensitivity of the proposed immunoassay offers great promises in providing a sensitive, specific, time saving, and potential method for detecting PA in clinical settings. ②The detection of monoamine neurotransmitters and their metabolites by high performance liquid chromatography:Based on before research, a high performance liquid chromatography for simultaneous separation and rapid determination of six monoamine neurotransmitters was development. The HPLC column was kromasil C18 (4.6x250 mm, size 5um). The mobile phase was a mixture of 90% methanol and 10% 0.1 mol/L natrium aceticum solution (v/v). The linear range of the method was 2-300 pmoL. The correlation coefficient of standard curve were higher than 0.9999, the average recoveries of monoamine neurotransmitters were from 95% to 102%. The method was a simple and rapid HPLC procedure, which allowed the large number of samples to be annlyzed within a suitable time. It was good for studying changes in neurotransmitter monoamine levels in different tissues of animal.③Optimization of a rapid and automated determination of salivary a-amylase:Based on before research, a faster and simpler method for determination of salivary a-amylase by flow injection spectrophotometry was optimized. The cycle of a sample using this method is only 34s. This system can be operated with a high precision, which can avoid big differences of determination caused by improper manual control.(2) Development of stress models:The stress (human or mice) model is one of the important method to study the relationship of stressor, behavior and neuroendocrine. Most of the studies have taken place in animal models, and human data is limited, but some studies of rodents and nonhuman primates can be extrapolated to human models. Buy the way of literature summary, we develop for animal and human stress model respectively:(1) mice model of stress (2) human stress model.(3) The construction of stress evaluation methods:In order to describe the influence of stress model on body, we develop the mice and human behavior evaluation method. Including mice behavior indicator:elevated cross maze, forced swimming test, feeding inhibition behavior, social behavior. Human behavior indicator:Stroop test, mental arithmetic evaluation, the cognitive regulation of eye movement.2. Effects of exercise on restrained stress-induced changes of the brain monoamine neurotransmitters, HDACs mRNA and behaviors in miceRestrained stress is a widely accepted model to study the long-term behavioral changes produced by the early life events. However, whether and how restrained isolation induce anxiety-like and social interaction behavior changes remain controversial, and if so, whether aerobic exercise can relieve it. Here we reported that 1-month-old mices subjected to individual isolations from their mother and nest for 2 h per day, increased anxiety and depressive-like behaviors tested in young adult (postnatal days 28-37) compared to normal reared controls. Furthermore, these behavioral changes were accompanied by altered brain monoamine neurotransmitter and HDACs mRNA expression. More importantly, two week aerobic exercise completely relieved restrained stress-induced anxiety, depressive-like and social interaction behaviors and restored the balance between excitatory and inhibitory activities to physiological levels. These findings indicate that restrained stress may produce social interaction change and anxiety-and depressive-like social interaction behaviors, and aerobic exercise may be an effective way to against negative emotion during development.3. Rapid determination of salivary a-amylase by Flow Injection Analysis for the research in children exercise stress responseThe aim of the study was to evaluate the applicability of flow injection spectrophotometry in the monitor of the potential biomarker, salivary a-amylase in acute exercise stress. Subjects were sixty healthy children who took a thirty minutes exercise test. To assess salivary a-amylase levels pre and post exercise, the participants provided four salivary samples (before the exercise test, at the end of the test and thirty minutes after the exercise test). It was verified that individual’s salivary a-amylase secretion was changed under exercise stress. It was concluded that flow injection spectrophotometry might be a reliable method for determination of salivary a-amylase in the assessment of exercise stress.4. Rapid determination of salivary a-amylase by Flow Injection Analysis for psychological stress response researchThe aim of the study was to assess the applicability of flow injection spectrophotometry in the monitor of the potential biomarker, salivary a-amylase in acute Psychological stress. Subjects were twelve healthy graduate students who took a ten minutes Stroop test and mental arithmetic test. To assess pre and post stress salivary α-amylase levels, the participants provided four salivary samples (ten minutes before the stress test, at the end of the each test and ten minutes after the third sample). It was verified that individual’s salivary a-amylase secretion was changed under physical or psychological stress, the method could detect salivaa-amylase activities conveniently and accurately. It was concluded that flow injection spectrophotometry might be a reliable method for determination of salivary a-amylase in the assessment of Psychological stress.5. Salivary alpha-amylase, heart-rate variability and behavior reaction in acute mild psychosocial stress and the impact of Tridimensional PersonalityRecent years have seen a growing interest in salivary a-amylase (sAA) as a non-invasive marker for sympathetic nervous system (SNS) activity. TPQ Personality includes 3 dimensions, defined as individual differences in implicit associative responses to mild stress stimuli. We examined whether these three dimensions were associated with patterns of Stroop and mental arimacimatics test and the counterpart sAA responses. Methods:From 24 healthy students, high and low novelty-seeking (NS) groups, high and low novelty-seeking (NS) groups, high and low Reward dependence (RD) groups high and low high and low Reward dependence (RD) groups, and high and low harm-avoidance (HA) groups were selected using scores on the Tridimensional Personality Questionnaire measuring temperament dimensions. Patterns of Stroop and mental arimacimatics stress test were presented, and sAA, HRV responses were measured as an autonomic index of stress responses. Stress responses to the two stimulus conditions were compared between groups. Results:LF/HF1 responses in the low NS, RD group were significantly greater than in the high NS, RD group before stress test were presented. Post of stress test and recovery of salivary a-amylase lever in the high RD group were significantly greater than in the low RD group for stimuli of all three valences. Number of trials of Stroop in high NS group were significantly greater than in the low NS group. Conclusions: suggesting that different dimensions of temperament may be associated with different patterns of Stress responses. Novelty seeking may be associated with susceptibility of preattentive perception.6. Stress reducedthe capacity of attention responses regulationCognitive regulation has been widely shown in the laboratory to be an effective way to alter the sport attention responses. We often fail to use Cognitive regulation strategies in high stress condition. In this research, visual search processing in sports situation of epee athletes and college undergraduates were analyzed. All participants first learned to practice epee situation task. Eye movement training directly followed where participants were taught to regulate responses to epee Attaque stimulus. Then, participants underwent a 3 min cold pressor stress or a control task before repeating the attack task using these newly acquired regulation skills. Salivary a-amylase and cortisol concentrations were assayed as neuroendocrine markers of stress response. The results showed that Top-ranking expert fencers fixated particularly on the trunk. In the first visual fixation stage, athletes collected more effective visual information. Although there was no group differences in arousal after the initial learning, eye movements of stressed collagestudentsparticipants showed declined attention regulation, which suggest that stress markedly impairs the visual cognitive regulation of new-formed skills affective responses under stress.