| ObjectiveBy detecting the gene polymorphism of MUC1-568 and HSP70-2-1267 in the gastric mucosa of patients who have Helicobacter pylori(Hp)correlated diseases(HPCD),controlled with the normal people,the relationships of disharmony syndrome between liver and stomach,damp-heat of spleen-stomach,spleen-qi deficiency syndrome and gene polymorphism of MUCI-568 and HSP70-2-1267 have been investigated.Also the susceptible genes of specific different syndromes of Chinese medicine have been discussed,in order to seek out some theories for the clinically integrated traditional and western medicine in the point view of gene polymorphism.Methods1.Source of casesThe pathological specimens of gastric mucosa were gathered from those outpatients or inpatients who visited the digestive department of the first affiliated hospital of Guangzhou University of Chinese Medicine,from May 2011 to October 2013,in which 99 cases of HPCG conforming to the inclusion criteria were included,72 cases with chronic gastritis,27 cases with peptic ulcer,30 cases with disharmony syndrome between liver and stomach while 27 Hp positive,37cases with damp-heat syndrome while 33 Hp positive,32 cases with spleen-qi deficiency syndrome while 26 negative,simultaneously 13 healthy people were recruited from Guangzhou University of Chinese Medicine while 2 people were Hp positive.All the general information of the subjects was inquired by the members of our research group,including clinical symptoms,then symptoms classification,grading,filling the clinical observation tables.2.Collection of pathologic specimensAll the subjects in different groups were examined by electronic gastroscopy,4 specimens collected in the greater gastric curvature or lesser curvature of the stomach antrum of every subject,at a distance(2 or 3 centimeters)from pylorus by biopsy forceps,and every specimen is about 0.3-0.4cmX0.3-0.4cm in size,one of which was detected by rapid urease tests,while one of which was fixed in 4%paraformaldehyde,then sent to pathology department in time in order to be paraffin-embedded,cut sections,stained with methylene blue and HE.The other two specimens were put in the two separated freezing tubes,after cryopreserved in the liquid nitrogen in a specific time,stored in the-80?low temperature refrigerator.When all the required cases of specimens were collected,the gene polymorphism test began.3.Observation and detection of indicatorsObserve the infections and the extent of infections for the gastric mucosa,the inflammation of the gastric mucosa,the active degree,or the pathological progression of different stages with microscope.Amplification and identification the gastric mucosa DNA of every subject by using PCR and electrophoresis,then detect the gene polymorphism of MUC1-568 and HSP70-2-1267 of every specimen by using Sanger double DNA chain termination direct sequencing method.Results1.General information comparisonGeneral information of different group including age,sex,disease entities and clinical course,has no statistically significant difference(P>0.05),so the general information of each group is comparable.2.Comparison for the distribution of the disease entityThe incidence of peptic ulcer of male is higher than female,and there is statistically significant difference(P<0.05).While the damp-heat syndrome is higher than spleen-qi deficiency syndrome in the peptic ulcer obviously,also there is statistically significant difference(P<0.05).3.Comparison for the infections and the extent of infections of HpThe incidence of Hp infection in the healthy volunteers is lower than that of HPCD(chronic inflammation and peptic ulcer),and there is statistically significant difference(P<0.05).While there is a tendency that chronic inflammation is higher than peptic ulcer,however no significant difference(P>0.05).For the infections extent of Hp,the healthy subjects are lower than the HPCD(chronic inflammation and peptic ulcer),also there is statistically significant difference(P<0.05),however there is no statistically significant difference between the chronic gastric inflammation and the peptic ulcer.For the detection rates of the three types of Chinese Medicine syndromes,there is a tendency that the cases of disharmony syndrome of liver and stomach are more than that of damp-heat syndrome,also it exists in the relationship between damp-heat syndrome and spleen-qi deficiency syndrome,however there is no statistically significant difference(P>0.05).For the infection extent,there is a tendency that the cases of damp-heat syndrome and spleen-qi syndrome are more than spleen-qi deficiency,while there is no statistically significant difference(P>0.05).4.Comparison for the clinical symptomsThe symptom of bitter taste is heavier than that of peptic ulcer,and there is statistically difference(P<0.05).For the symptom of anorexia,the cases of spleen-qi deficiency syndrome are more than that of disharmony syndrome of liver and stomach or damp-heat syndrome of spleen and stomach.For the symptom of nausea,the cases of spleen-qi deficiency symptom are more than that of damp-heat syndrome and disharmony syndrome between liver and stomach.For the symptoms of bitter taste,the damp-heat syndrome of spleen and stomach are more than that of disharmony syndrome of liver and stomach or spleen-qi deficiency syndrome,also there is statistically significant difference(P<0.05).5.Comparison for the extent and activity of inflammation,the pathological progression stages of gastric mucosaThere is a tendency that the cases of the peptic ulcer group are more than that of chronic inflammation for all the activity and extent of inflammation,the degree of atrophy,however there is no statistically significant difference(P>0.05),while there is no difference between the intestinal metaplasia and dysplasia group.There is a tendency that the cases of disharmony syndrome of liver and stomach or the damp-heat syndrome of spleen and stomach are more than that of spleen-qi deficiency syndrome for the degree of inflammation and atrophy,however there is no statistically significant difference(P>0.05).6.Relationship of gene polymorphism of MUC1-568?HSP70-2-1267 and HPCDThere is no statistically difference for the genetype frequency of MUC1-568 in the HPCD and healthy volunteers(P>0.05).There is a tendency that the cases of peptic ulcer are more than that of healthy volunteers or chronic gastric inflammation for the HSP70-2-1267GG genetype frequency in the HPCD,and there is no statistically difference for the rest of the genetype frequency in different groups(P>0.05).7.Relationship between the genetype polymorphism of MUC1-568?HSP70-2-1267 and Hp inflammationThe tendency that genetype frequencies of AG in MUC1-568 in Hp positive are more than that of Hp negative exists,however there is no statistically difference for the rest of the genetype frequencies in the two groups(P>0.05).There is no statistically difference for the genetype frequencies of MUC1-568 in different groups(P>0.05).There is also no statistically significant difference for the genetype frequency of HSP70-2-1267 between the Hp positive group and Hp negative group(P>0.05).8.Relationship of genetype polymorphism of MUC1-568?HSP70-2-1267,the extent and activity of inflammation,the pathological progression stages of gastric mucosaThere is no statistically difference for the extent and activity of inflammation,the pathological progression stages for the genetype frequency of MUC1-568(P>0.05).GA genetype of HSP70-2-1267 possibly is related to the intestinal metaplasia and hyperplasiathe of gastric mucosa.9.Relationship between gene polymorphism of MUC1-568?HSP70-2-1267 and three types of Chinese Medicine syndromeAmong the genetype frequency of MUC1-568AG in the spleen-qi deficiency syndrome,disharmony syndrome of liver and stomach,the damp-heat syndrome of spleen and stomach and the healthy volunteers,there is a tendency that the genetype frequency that decrease progressively in those four groups,however there is no statistically significant difference(P>0.05).Also there is a same tendency in the genetype frequency of HSP70-2-1267AG,but no statistically significant difference(P>0.05).ConclusionTo investigate the relationship between the gene polymorphism of MUC1-56,HSP70-2-1267 and HPCD,rapid urease test,methylane blue stain,HE pathological stain and Sanger double DNA chain termination direct sequencing method were used.The conclusions are following:1.The incidence of peptic ulcer in male is higher than that of female,and the cases of damp-heat syndrome are more than that of spleen-qi deficiency syndrome in the peptic ulcer.There is a tendency that the extent,activity and atrophy of inflammation in the gastric mucosa of peptic ulcer are heavier than that of chronic gastric inflammation group.The degree of inflammation and atrophy of gastric mucosa in Shi syndrome including disharmony syndrome of liver and stomach and the damp-heat syndrome of spleen and stomach is heavier than that of spleen-qi deficiency syndrome.2.The GA genetype of HSP70-2-1267 is corraleted with intestinal metaplasia and displasia of gastric mucosa.Also the GG genetype of HSP70-2-1267 may be the susceptible genetype of peptic ulcer,AG genetype of MUC1-568 is possibly related to the syndrome of spleen-qi deficiency,and AA genetype of HSP70-2-1267 is possibly related to the damp-heat syndrome,while the gene polymorphism of MUC 1-568 may not have any correlation of HPCD(chronic gastric inflammation and peptic ulcer)or Hp. |
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