| Objective:To clarify the dynamic evolution of the heat and blood stasis machine in acute cerebral hemorrhage.By finding the relationship between the pathogenesis and laboratory indicators,and the related plasma proteome,we want to lay a foundation for the definition of the pathogenesis and clinical blood biomarkers.Method:1.Based on the "973" project of TCM in 2006,the "Study on Inheritance and Innovation of Etiology and Pathogenesis of TCM" subtopic:"Discuss the rule of the internal hard-to-treat disease and the molecular basis of its syndrome nature from the theory of "heat and blood stasis".The clinical data of 511 acute cases of cerebral hemorrhage collected during the period from 2007 to 2010 in neurology departments of 15 tertiary hospitals such as Jiangsu Provincial Hospital,Nanjing Chinese Medicine Hospital,and Zhongda Hospital,using EpiData3.2 Chinese data entry,SPSS 22.0 statistical analysis software for six types of laboratory indicators(blood pressure,fasting blood glucose,blood lipids,hemorheology,coagulation,cranial imaging(CT)),two kinds of heat and blood stasis score results(famous medical experience The descriptive analysis of the measurement model and the measurement method of the heat and blood stasis unit scale,the linear correlation and the rank correlation analysis between the heat and blood stasis score(famous medical experience measurement model)and various laboratory indicators.2.Department of Neurology,Department of Neurosurgery,Jiangsu Provincial Hospital of Traditional Chinese Medicine,Binjiang Hospital of Zhenjiang,China,collected from 2013 to 2015,based on the National Natural Science Foundation of China's "Dynamic Measurement of Acute Cerebral Hemorrhage and Heat and Blood Stasis Units and the Study of Their Biomarkers" The blood samples and data of 168 patients with acute cerebral hemorrhage were used in the acute phase of hemorrhagic stroke developed by the expert group of the "973 mechanism of heat and blood stasis due to the mechanism and molecular basis of the disease in the department of internal medicine" of Nanjing University of Chinese Medicine.Quantitative diagnostic criteria for these syndromes were performed on these patients and were divided into cerebral hemorrhagic heat and blood stasis type and non-heat and blood stasis type,and then paired according to sex,age,course of disease,bleeding site,and heat and blood stasis score to obtain 40 pairs of experimental samples(40 in the acute phase of heat and blood stasis,40 in the heat and blood stasis recovery phase,40 in the non-heat and blood stasis acute phase,and 40 in the nonheat and blood stasisc recovery phase).Blood samples of these 40 experimental samples were performed using the SWATH method.Quantitative proteomics studies using the EpiData3.1 Chinese version(provided by the World Health Organization)to establish a clinical data database and SPSS 22.0 statistical analysis software for data analysis and calculations At the same time,the heat and blood stasis score("quantitative diagnostic criteria for pathogenesis of hemorrhagic stroke"),the correlation between the heat and blood stasis score at the initial diagnosis and the protein information at the corresponding period,P<0.05,Pearson correlation coefficient |r|Proteins with>0.3 were subjected to bioinfonnatics analysis of the UniProt and Metascape databases.Results:1.Retention between the heat and blood stasis and the laboratory indicatorsThere was a strong positive correlation between the first day and the first day of hospitalization with the heat and blood stasis score.Hemorheology(erythrocyte aggregation index),cranial imaging(CT)(total cerebral hemorrhage);On the 11th day after the initial diagnosis and admission,both were negatively correlated with heat and blood stasis scores with hemorheology(whole blood viscosity value 30,whole blood viscosity value 5,plasma viscosity value 100),coagulation(plasma fibrin)There was no correlation at the time of initial diagnosis;there was a weak positively correlated fasting blood glucose at 11 days after admission;no correlation was found at the time of initial diagnosis,and weak negatively related lipids(triglycerides)and blood were present at 11 days after admission.Rheology(whole blood viscosity:200);Weakly positive correlation at first visit,no related lipids(low-density lipoprotein)11 days after admission;weak negative correlation at first visit,11 days after admission See correlation for hemorheology(whole blood viscosity value 1,erythrocyte sedimentation rate),coagulation(activated partial thromboplastin time),and blood pressure(not associated with heat and blood stasis score)at the first visit and on the 11th day after admission.Systolic,diastolic,lipids(cholesterol,high)Density lipoprotein,apolipoprotein A,apolipoprotein B,lipoprotein a),coagulation(prothrombin time,thrombin time),cranial imaging(CT)(brain edema grade).2.The evolution law of heat and blood stasis2.1 According to the analysis results obtained from the "famous physician experience measurement model score" we found that:(1)Almost all patients have different degrees of symptoms of heat and blood stasis,but according to the experience of famous doctors,half of the patients with AICH met the diagnostic criteria for heat and blood stasis,manifested as A typical heat syndrome,therefore,heat and blood stasis pathogenesis is an important pathological feature of acute cerebral hemorrhage,but the difference between light and shade.(2)On the third day after admission,it is often the third day after the onset of illness.The heat and blood stasis score is the highest and the symptoms are the most obvious.It reflects the extreme performance of AICH disease and the condition is serious.The hotness level of the heat and blood stasis-associated symptoms collected at the time of admission meets the diagnostic criteria for the heat and blood stasis.Therefore,whether the presence of heat and blood stasis reflects the severity of the disease in the acute phase of cerebral hemorrhage to a certain extent can be used to monitor the disease.Progress of the disease.(3)After the peak period,the level of heat and blood stasis did not change sharply,but it also showed a gradual decline,and overall it was faster than before the disease.No second peak of heat and blood stasis was observed,which may be consistent with the characteristics of AICH's own course of change,which is a pathological reaction that occurs within a short period of time.The condition is severe and reflects the severity of the disease;The situation did not collect information,and the possibility of secondary hemorrhage in cerebral hemorrhage could not be known for the development of the development of the sputum heat and blood stasis during the entire AICH course,which needs to be further studied in the future.It can be seen that identifying the presence and degree of heat and blood stasis in the early stage of AICH can early monitor the progress of the disease and predict the prognosis.It is especially important to provide a diagnostic basis for early individualized interventions in Chinese medicine,which will greatly reduce the disease.Mortality and disability rates.2.2 According to the results obtained from the use of a helium calorimeter,"measurement method of the heat and blood stasis unit scale," we found that:(1)Most patients have any of the four phlegm heat variables,which is a testimony to the heat and blood stasis machine.With multiple dimensions,it divides the heat and blood stasis machine according to traditional Chinese medicine theory and clinical manifestations into several pathogenesis differentiation factors(the heat and blood stasis machine may include four changes of real heat,deficiency heat,blood stasis and consciousness),and carries out Objectively quantified measurement will make the pathogenesis differentiation more accurate.According to the theory of hemorrhagic stroke and heat and blood stasis caused by Professor Zhou Zhongying of the Chinese medicine master,different patients and the disease period "blood stasis" and "heat" are of different severity,and they are transformed into each other.In the early stage,the heat was heavy and the middle heat was heavy.In the later period,the heat was heavy.The establishment of this specific division method reduces the difficulty of judging the pathogenesis of paralyzed heat and blood stasis,and provides a reliable tool for the study of the microscopic mechanism of combining clinical testing indicators.(2)By analyzing the four latent variables independently,we can find that the real heat syndrome and blood stasis syndrome are long-term indications with obvious specificity,suggesting that the stasis heat syndrome is more empirical,as a cerebrovascular disease The first manifestation of consciousness,from the beginning of the disease to the late stage of the disease,is the difference between the four latent variables,the most obvious manifestation of acute cerebral hemorrhage acute disorders,but once the early days,consciousness recovery faster.3.Retention between the heat and blood stasis and plasma proteins3.1 Correlation Analysis:Pearson(r)was between-0.507 and 0.521,there were 84 proteins with P<0.05;there were 55 proteins with correlation coefficient | r |>0.3:P09871,Q14008,P02763,P08697,P01834,P01596;P01598,P01011,P01860,P15169,P69905,P51812,Q562R1,P02649,P13798,P18428,P01825,P06331,P01593,P01594,P01599,P01600,P01607,P01608,P01609,P01610,P80362,Q9BWK5,P60709,P63261,Q8NCM8,P68871,P05452,P03952,P05165,P00746,P00488,P02774,P00751,P05155,P07225,P01024,P00736,P06396,075636,P02042,P02749,P02775,P00915,P02753,P01620;P01622;P01623;P04206,P02750,P02787,P02790,075449,P01625;P06312;P06313;P06314,P36955,Q06033,P02747,P04003,P00740,Q9BSC4,075882,PODJI8,Q6GMV1,P27169,of which 31 proteins are positively correlated and 24 negatively correlated.3.2 Bioinformatics analysis:In the acute phase of cerebral hemorrhage,heat and blood stasis-associated plasma proteins mainly exist extracellularly or originate from secretion,and positively related proteins are mainly present in the plasma membrane and cytoskeleton,and very few are scattered in the nucleus,endoplasmic reticulum,or cell fluid.In mitochondria,only a small portion of negatively related proteins are scattered in the plasma membrane,cytoskeleton,nucleus,or endoplasmic reticulum.The plasma proteins associated with heat and blood stasis in the acute phase of cerebral hemorrhage come from different families.Both the positive and negative correlations belong to the peptidase S1 family of proteins.Among positively related proteins,the globin family,actin family,and peptidases Each of the S1 family has more than one protein subordinate.In the negatively related protein,the peptidase S1 family and the serpin family each have 4 protein subordinates,and the cardinomycin superfamily and the Lipocalin family each have 2 protein subordinates.About one-third of the plasma proteins associated with the heat and blood stasis machine in the acute phase of cerebral hemorrhage have the function of regulating metal ion binding.Both positive and negative correlations have ATP binding,actin binding,metal ion binding,and molecular functions.Antigen binding,enzyme binding,endopeptidase inhibitor activity,chemotaxis activity,serine-type endopeptidase activity,carbohydrate binding,same protein binding,lipid binding,and a third of positively related proteins have a serine type The function of the peptidase activity,the rest of the negatively related proteins,is mainly related to the functions of serine-type endopeptidase activity,endopeptidase inhibitor activity,heparin binding,the same protein binding,ATP binding,lipid binding and carbohydrate binding.Both positive and negative plasma proteins associated with heat and blood stasis scores were enriched into 8 biological processes,of which the same biological processes had only lipid metabolism.According to the degree of participation from high to low,the biological processes specific to positively correlated proteins were:Complement and coagulation cascade,hemostasis,exocytosis,coagulation,red blood cell function,protein processing,phagocytosis,and biological processes specific to negatively related proteins are:acute inflammatory response,zymogen activation,acute phase response,and NABA MATRISOME-related,NABA ECM modulators,receptor binding and uptake ligands,immune effects.Conclusion:1.In clinical laboratory indicators,there must be associated with heat and blood stasis,blood sugar,low-density lipoprotein,red blood cell aggregation index,activated partial prothrombin time,total cerebral hemorrhage,may be related to triglyceride,lipoprotein a.Erythrocyte sedimentation rate,plasma fibrinogen,unrelated to blood pressure,cholesterol,high density lipoprotein,apolipoprotein A,apolipoprotein B,whole blood viscosity value 200,whole blood viscosity value 30,whole blood viscosity value 5,whole blood viscosity Value 1,plasma viscosity value 100,prothrombin time,thrombin time,brain edema grade.2.Almost all patients have different degrees of symptoms of heat and blood stasis,and the most severe heat and blood stasis occurs on the third day after the onset.Most patients have any of the four types of heat and blood stasis latent variables.Dimensions,real sthenic fever syndrome and blood stasis syndrome have a long time in the course of acute disease.3.There were 55 plasma proteins associated with heat and blood stasis in the acute phase of cerebral hemorrhage(P<0.05,correlation coefficient | r | ? 0.3),of which 31 were positively correlated proteins:Q14008,P06396,P60709;P63261,Q06033,Q562R1,P01620;P01622;P01623;P04206,P00488,P01860,P02775,P03952,Q9BSC4,P01825;P06331,075449,P07225,Q6GMV1,P01834,P01596;P01598,P02042,P68871,P04003,P13798,P02787,P00751,P69905,P00915,P01165,075882,P01593;P01594;P01599;P01600;P 01607;P01608;P01609;P01610;P80362,P01625;P06312;P06313;P06314,P01024,P27169.24 negatively related proteins:P09871,P00736,Q9BWK5,P36955,P51812,P02753,P00746,P01011,P02749,P05452,P00740,P18428,P02790,P05155,P0DJI8,P02763,P08697,075636,Q8NCM8,P02747,P02649,P02750,P02774,P15169.4.These plasma proteins mainly exist in extracellular or secretory origin,basically from different families,both positive and negative related parts belong to the peptidase S1 family of proteins,about one-third have the function of regulating metal ion binding,Both negative and related components possess ATP binding,actin binding,metal ion binding,antigen binding,enzyme binding,endopeptidase inhibitor activity,chemotactic activity,serine type endopeptidase activity,carbohydrate binding,the same protein binding,lipid binding.5.Eight biological processes that enriched positive and negative plasma proteins associated with heat and blood stasis scores were obtained.The common biological processes are lipid metabolism,and biological processes unique to positively related proteins:complement and coagulation cascade reactions.Hemostasis,exocytosis,coagulation,erythrocyte function,protein processing,phagocytosis,and biological processes specific to negatively related proteins:acute inflammatory response,zymogen activation,acute phase response,NABA MATRISOME-related,NABA ECM modulators,Body binding and uptake of ligands,immune effects.Once again verified that lipid metabolism is the pathological mechanism of cerebral hemorrhage,it also confirmed to a certain extent that the microscopic mechanism of the heat and blood stasis machine in the acute phase of cerebral hemorrhage is related to the coagulation process and inflammatory reaction,but this study found that the pathogenesis may be more presented as the coagulation process,and the inflammatory response plays an opposite role at this time. |
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