The Clinic Research And Study On Difference Gene Expression Bioinformatics Of Chronic Gastritis Spleen Deficiency Syndrome (Gastricism And Malabsorb Subtype)

BackgroundChinese traditional medicine presume that syndrome is the pharmacolog summarize of the cause,location,nature,tendency of disease in the stage of disease process. Syndrome research is the hot spot in traditional Chinese medicine. Many researchers dedicate in syndrome modernization research for decades. They use many modern technique and method to reveal the mystery of syndrome in biochemistry, physiology,immunology,ultrastructure,elements aspect。They get some achievements and confirmed that the Syndrome have pathophysiology foundation. But. weak specificalness is the important cause of cursing syndrome to make great advancement. Many experiments consider that there is no study means to match the complexity and entirety of Chinese medical science syndrome.In recent years, accompanying with the development of genomics, modern pathogenesis consider that the disease is the interaction result between many correlated genes and interior or external environment. Gene chip can parallel analysis massive genes in the same time and take mass information filter. So it is to be used in mechanism,classify,diagnosis,prediction of disease. Many scholars believe that gene chips research method is coincidence with characteristic of the Chinese traditional medicine syndrome, They think gene chip can provide drastic technique support in approaching the essence and objective research of syndrome. Many researchers begin to use gene chip to study syndrome and confirm that different syndrome have different gene expression profile. chronic gastritis is chronic inflammation of gastric mucosa, the patient was devided in several types according to their clinic symptom-complex difference, they can get better therapy result chronic gastritis spleen deficiency syndrome (gastricism and malabsorb subtype ) is one type of this disease. In our early research ,we find the spleen deficiency syndrome patient gene expression profile is different from health adult and piwei damp-heat syndrome of chronic gastritisThe spleen deficiency syndrome patient/s genes are down-regulated in protein synthesis,immune function,energy and substance metabolism,gene transcription. modern medicine research show that some great disease such as tumor,diabetes,cardiovascular disease is correlated with some definite genes and they have great repeatability, some diagnose gene chip fs already used in clinic. But the syndrome of chinese medical science is a different science system with medical disease. The syndrome of chinese medical science have the feature of abstractive, fuzziness, non- quantitive. There are many factor to participate and regulate in syndrome, so it is more complicate than disease. Accordingly, the repeatability of Chinese medical science syndrome differential expression gene is called in question. Wether the differential expression genes we get from the early research can represent the diagnostic criteria / s differential expression genes, How about the relationship between symptoms of syndrome and differential expression genes , and the feature of gene expression profile of spleen deficiency syndrome should to be studied deeply. According to this study , we want to act out the pathophysiological hypothesis of spleen deficiency syndrome different gene expression profile and develope the diagnostic criteria of spleen deficiency syndrome (gastricism and malabsorb subtype )Methods and Result1.Subjects collectionMethod: case of a particular disease is to be in line with chronic gastritis diagnostic criteria (clinical,gastroscop,pathologic diagnosis), consistent with spleen deficiency syndrome/ piwei damp-heat syndrome diagnostic criteria, rule out alimentary tract structural disease and other system severe disease. Healthy manResult: the group of spleen deficiency syndrome was 21 cases, and that of piwei damp-heat syndrome was 28 cases, and that of normal people was 14 cases. Each subject rings clamp gastric mucosa for histopathology and gene chip experiment After filtered by RNA extraction, choose 8 cases each group for gene chip experiment 2. clinical data analysisMethod: analysis the patients clinical symptom and pathology, get information about chronic gastritis clinical symptom and pathology relationship, the different clinical symptom between spleen deficiency group and the piwei damp-heat group, classify spleen deficiency degree by clinical symptom, search significant symptom for spleen deficiency degree classification.Result: HP of yellowish greasy tongue is higher than that of thin and whitish fur (P=0.015),There is no statistically significance between HP and gender, course of disease, pathology, but accompany with course of disease ;lasting and inflammation aggravation, the infection rate of HP is increased, it is perhaps relative to sample size. in addition, the eldest of body of tongue have dependability with loose stool, it has statistically significance (tau_b=0.384 P=0.007)There have no statistically significance in male female ratio, course of disease, pathology degree between spleen deficiency group and the piwei damp-heat group. The symptom of painful abdominal mass, anorexia, less food are higher than that of piwei damp-heat group(P=0.007,P=0.005,P=0.007). HP infection and abdominal pain is higher in piwei damp-heat group(P=0.019,P=0.007).The frequency of occurrence of spleen deficiency syndrome symptoms exceed 90 percent are pale tongue with indentation, abdominal distention after eating, tired and debilitation, anorexia, the symptom of loose stool occurrence is only 47.6 percent, it shows that loose stool appears in some degree spleen deficiency, extracting the occurrence of symptom above 45 percent to degree the spleen deficiency syndrome. It shows that the score of main syndrome is coincidence with the score of whole body syndrome. The most patients of spleen asthenic syndrome is lightly or midrange spleen asthenic.The symptom of loose stool and weary spirit have statistical significance in classification in spleen asthenic syndrome degree (P=0.017, P=0.030). Although fat body of tongue and wasting have statistical significance in classification in spleen asthenic syndrome degree, but the occurrence of this two symptoms is highest in heavy degree spleen deficiency syndrome group. It. shows that the course of spleen asthenic syndrome develop from digestive canal to general symptom. The symptom of insomnia has no statistical significance with sex ,but has statistical significance with body weight decrease(tau_b=0.748 P=0.001).3. Gene chip data analysis Method: the gastric mucosa was collected under the help of gastroscope and stored in liquid nitrogen. The gene expression of gastric mucosa was examined by BiostarH-140s. there are two group gene chips, one is spleen deficiency syndrome patient via healthy people(Group AB), the other group is spleen deficiency syndrome patient via piwei damp-heat patient (Group AC). The different gene is choosen by cyS/cy3 ratios and t-test, then take analysis of gene bioinformatics. Cluster the patients by filtered gene, according to the symptoms of the patients, to find the relationship between the syndrome with the gene expression. Compared two experiments different gene, Sum up the; feature of spleen deficiency syndrome gene expression profile.Result: There are eight gene chips in every group. There are 83genes filtered by cyS/cy3 ratios(l. Stimes)in group AB. There are 24 genes up-regulated , 51(61.4%)genes down-regulated. These genes is relative to protein synthesis and metabolism, genetic transcription, immunization, energy and nutrient substance metabolism, cytoskeleton. There are 12 genes filtered from 83 genes by t-test(P＜0.05).294 genes are filtered by cy5/cy3 ratios(1.5times)in group AC. 198(67.3%) genes are down-regulated, 83 (28.2%) genes are up-regulated. These genes is relative to DNA repair, protein synthesis and metabolism, energy and nutrient substance metabolism, immunization, cytoskeleton and cell cycle, signal transduction. There are 109 genes filtered from 294 genes by t-test(P＜0.05) 85 (77.98%) genes are down-regulated, 24 (22.02%) genes are up-regulated. AB group patients is clustered by all filtered genes(cyS/cy3 ratio). From the cluster result, we find the patient whose general symptom score is high(13,13,17,17)are obviously clustered in one group. The patient whose general symptom score is low is far from the other patient. The high score people have the symptom such as loose stool, fat tongue with indentation, the lower score patient have normal stool or dry stool, loose stool and fat tongue with indentation are the symptom of nutrient substance malabsorption. The ratio of genes especially correlated with nutrient substance absorb, energy metabolism and protein synthesis are down-regulated more obviously in the patient who have loose stool and fat tongue with indentation So we presume that some symptoms have its own genes background, loose stool and fat tongue is the key symptom in spleen deficiency syndrome classification.Bioinformation of some major difference genes: RPS28,RPS29,RPS20,RPL37,RPL35,RPL6,RPL23AP7,RPL22,RPL27,MRPL18,CGI-94,RPL18A,RPS7,RPS9,MRPL18,MRPL51,MRPS6 are take part in protein synthase, AKRIC1 and SLC13A4 are relative to bile acid transport, ECH1 administrates fatty acid metabolism, SLC38AI and SLCIAI is involved in amino acid transport, ARL61P5 regulates concentration of taurocholic acid and glumatic acid inside the cell. The gene relative about energy metabolism is LOC57149, CGI-69, ATP6V1G1, COX8A, PRDX3,CYP20A1 and NDUFB2. KRT19, ADD3, CAPZA2 and RCSD1 which are relative to cystoskeleton are down-regulated. SLOOP participates endothelial cell migration up-regulated. CENPH and SMS participate cell division and proliferation down-regulated.Conclusion1. chronic gastritis spleen deficiency syndrome (gastricism and malabsorb subtype) has its own different gene expression profile, the genes function include protein synthesis and metabolism, genetic transcription, immunization,, energy and nutrient substance metabolism, cytoskeleton. The difference genes in two chip experiment are coincident in the tendency of up or down-regulated and gene classification. It shows that diagnostic criteria is suitable, stability and confined specificity of syndrome. But complete replication genes are less, except foe considering about individual difference, time and space difference, two syndrome, light and heavy press of spleen deficiency should be considered.2. Different symptom has its own gene expression profile, clinical exo-syndrome of syndrome and modern biology are two aspect of one unity. To analyze the patient clinical data, we discover frequency of secondary symptom weary and debll, is ahead, and the about chondrosome energy metabolism is down-regulated, whether bring this symptom into main syndrome should be discussed. Frequency of loose stool is only 47%, the ratio in middle and heavy spleen deficiency patient is obviously high, fat tongue shows direct ratio with spleen deficiency degree, and has dependability with loose stool. The ratio of genes especially correlated with nutrient substance absorb, energy metabolism and protein synthesis are down-regulated more obviously in the patient who have loose stool and fat tongue with indentation, these patients who have high symptom score are clustered in one group. so we presume fat tongue and loose stool be reference symptom of degree classification.3. Because syndrome is not identify and fixed quantity, relative to multi-system, combination differentiation of symptoms and signs, it is invilable to research symptom in wide-bound. We should choose one bound patient with special symptom to study4. It is one point of view to study syndrome ,its outcome could be a supply to other indicatrix, it can not displace clinical diagnosis. This research belongs to initial stage, there are many question to be solve, such as; how to analyze the gene chip data further deeply, how to mix un-quantitative clinical symptom message, patho-physiology and gene bioinformatics together better.