1.The Study Of Protective Effects On Cerebral Ische Mia-reperfusion Injury Of A New MLIF Analogue Replacement By Cysteine In Rats 2. Analysis Of The Normal Values Of Pulmonary Function By Impulse Oscillometry In Healthy Adults In Lanzhou

BackgroundIschemic cerebrovascular disease（ICVD） is one of the major diseases that are serious and harmful to the human health and survival. It is considered now that the acute inflammation is the critical factor to the process of the secondary reperfusion injury. There includes a large cytokines released, the high expression of adhesion molecules. White blood cells and other immune cells are activated to release a variety of inflammatory mediators such as cytokines including TNF-α, IL-1β, IL-6, IL-8, chemokines and adhesion molecules. TNF-α is considered to be the key factor in the whole inflammatory network and is the medium of the beginning of the systemic inflammatory responses. During the period of ischemia and reperfusion, there is the production of free radical increased, the remove of free radical declined and lipid peroxidation enhanced significantly at the same time. Nerve cells will be damaged severely. Therefore, if the inflammation reduced and the free radical chain cut, it is very important to protect the function of nerves after ischemia-reperfusion. Monocyte locomotion inhibitory factor（MLIF） is a polypeptide that is an anti-inflammatory response. MLIF inhibits the movement of human monocytes and polymorphonuclear leukocytes, suppresses the inflammatory response and escapes the host immune. At the same time, it also inhibits the secretion of cytokines TNF-α, IL-1β and the expression of the corresponding receptors. It affects the whole process of inflammatory reactions. There have been shown that the group of atoms responsible for the action of peptide Met-Gln-Cys-Asn-Ser（MLIF）is the carboxyl-terminal group Cys-Asn-Ser in the studies. The N-terminal of MLIF active groups Cys-Asn-Ser will be modified in this experiment. It will be understood that how does the new analogue of MLIF affect the changes of nerves function and impact the impression of TNF-α, IL-1β. It will be hypothesized the pharmacological mechanism of the new MLIF analogue. So it will provide the theoretical basis for the clinical study and will give the benefit to the patients with ischemic cerebrovascular disease in the future. Objective. There has two parts in this study. In order to increase the anti-inflammatory activity of MLIF, in the first part, there were designed and synthesized one new analog（Cys-Cys-Asn-Ser） of MLIF and the N-terminal of MLIF Cys-Asn-Ser by solid phase synthesis. The second part is that comparing with the N-terminal of MLIF Cys-Asn-Ser, we studied the pharmacological activity and potentially mechanism of the new analog of MLIF on cerebral ischemia-reperfusion injury by set up MCAO in rats. Methods1. In this study, we transformed and synthesized the new analog（Cys-Cys-Asn-Ser） and the N-terminal of MLIF Cys-Asn-Ser by solid phase synthesis. For desalination treatment, Sephadex G10 gel column was used. At the same time, we used HPLC to purify and needed Mass Spectrometry to identify the structure of the two peptides.2. An intraluminal monofilament was applied to blockade of the MCA（Middle cerebral artery）and then the focal cerebral ischemia was induced in male SD（Sprague Dawley）rats. For observing the role of this peptide on focal cerebral ischemia-reperfusion injury in SD rats, the neurological functions was judged by Longa’s method. After 24 hours reperfusion, with TTC staining technique we still measured the infarction size of cerebral tissue.3. After 24 hour reperfusion in SD rats, the level of TNF-α and IL-1β were detected in brain tissue homogenate for evaluated the anti-inflammatory activity and the function of neuroprotective effect of this peptide. Results1. There were verificated the chemical structure of the new analog（Cys-Cys-Asn-Ser） of MLIF and the micro-molecule polypeptide Cys-Asn-Ser by HPLC and MS. The output of Cys-Cys-Asn-Ser was 40.5% and the yield of Cys-Asn-Ser was 61.2%. Their puritys are more than 95%.2. After MCAO in SD rats, there was showed distinct neurological deficits in the ischemia-reperfusion group rats; a serious infarction was displayed in the ischemic brain tissue. Pretreated and treated with these peptides after MCAO in rats, the size of infarction was reduced and the neurological deficits were taken a turn for the better. Compared with the N-terminal of MLIF Cys-Asn-Ser, there was the consequence to imply that the pharmacological activity of new analog（Cys-Cys-Asn-Ser） has been enhanced3. In this study, it was concluded that TNF-α and IL-1β were increased in model group and the expression of TNF-α and IL-1β were inhibited by the new analog Cys-Cys-Asn-Ser and the micro-molecule polypeptide Cys-Asn-Ser. Compared with the N-terminal of MLIF Cys-Asn-Ser, there has been extremely increased the anti-inflammatory activity of the new analog Cys-Cys-Asn-Ser. ConclusionOn the basis of these findings, we eventually concluded that the anti-inflammatory activity of Cys-Cys-Asn-Ser has been improved in comparing with the pharmacophore（Cys-Asn-Ser） of MLIF. This new analog Cys-Cys-Asn-Ser protects the nerves function on Cerebral Ischemia-Reperfusion Injury.Background and ObjectiveThe respiratory system disease is a common and frequently-occurring illness. It has been a major public health implication because it causes serious damage to the people’s health. Not only it is high incidence, many diseases are insidious onset, but also it damages seriously pulmonary function and it is high morbidity, which has caused huge losses to the society and national economy. So it is very important of the early diagnosis, early treatment and early prevention on the respiratory system disease. In the examination methods of respiratory disease, we have more and more pay attention to the determination of lung function. But the regular examination of lung function always take the breathing of subjects as a signal source, it is inevitably affected by the subjects and will be not conducted especially on the old, the young, the weak and the consciousness and so on. The prominent advantage of IOS is that the subjects can breath quietly, the operation is very convenient and it is applicable widely. So IOS is very common in the clinic now. The normal values of the determination of lung function are usually affected by the subjects’ gender, age, height, weight and other factors such as the race, the physical activity, the type of work, the living environment and smoking or not. It is the necessary measure to improve the accurate diagnosis for selecting the right reference values. But there don’t have the uniform reference values and the normal values in China. The reference values that we always have adopted are provided by the abroad（mainly European）.It is not accord with Chinese practical. The standard of IOS is inconsistent in the different domestic region. So it is not conducive to make an accurate diagnosis. Therefore it is particularly important to establish the predictive equations of IOS in China. Lanzhou is located in the northwest of China. Because of the special geography and as a industrial city, it is necessary to establish the normal values and the predictive equations in Lanzhou area. Our results will be provided the reference to the clinical doctors especially for the respiratory doctors in Lanzhou and in the other five provinces of the northwest and provided a clue for the clinical doctors in the other domestic areas. It will be the foundation to build our ownand uniform the normal values and the predictive equations early in China. Materials and MethodsA total of 920 healthy adults who are over the age 18 and the pre-surgery patients who don’t have cardiopulmonary and airway disease were enrolled in this study. The subjects were enrolled in the Second Hospital of Lanzhou University between February 2008 and December 2011.There are 450 males and 470 females. The subjects were measured by the determination of IOS which standard are totally according to the recommendation from European Respiratory Society（ERS）.The subjects have not been taken medicines such as beta-2 agonists, theophyllines, cholinergic receptor antagonists, hormones, anti-allergics, adrenergic and so on before 24 hours. Respiratory impedance（Zrs）, respiratory resistance（R5 and R20）,resonant frequency（Fres）,reactance（X5）,central and peripheral resistance（Rc,Rp） and so on are measured with Jaeger Master Screen IOS instruments. All statistical analysis was processed by SPSS 17.0. Results1. IOS parametric such as Zrs, Fres, R₅, R₂0, X₅, Rc and Rp are not normally distributed. 2. In the parametric of our study such as Zrs, Fres, R₅, R₂0, X₅, Rc and Rp, their values have difference with the proposed value of foreigner. The two paired test is significant different（P<0.001）. 3. The predictive equations of IOS main parametric have been set up respectively in both male and female healthy adults in Lanzhou. 4. The average and the 95% confidence interval of Fres are greater than 10 Hz which always used at home and abroad, even greater than 15 Hz.There is no significant different from male and female. 5. According established the predictive equations of IOS main parametric, R is related to the height and the weight, especially the height. There is a negative correlation. But it is a positive correlation with the weight. 6. Considering the ages（within the limits of 18 and 80） in our study, there is no obvious change in the parametric of IOS（except X₅） as the growth of age. X₅ is a negative correlation with the age. ConclusionThe reference values and predictive equations of IOS for healthy adults in Lanzhou are different from those inland and abroad. It should be set up our own reference values and predictive equations.