Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis

AIM:To assess expression of matrix metalloproteinases 2(MMP2)and MMP9 in gastric cancer,superficial gastritis and normal mucosa,and to measure metalloproteinase activity.METHODS:MMP2 and MMP9 mRNA expression was determined by quantitative real-time polymerase chain reaction.Normalization was carried out using three different factors.Proteins were analyzed by quantitative gelatin zymography(qGZ).RESULTS:18S ribosomal RNA(18SRNA)was very highly expressed,while hypoxanthine ribosyltransferase-1(HPRT-1)was moderately expressed.MMP2 was highly expressed,while MMP9 was not detected or lowly expressed in normal tissues,moderately or highly expressed in gastritis and highly expressed in cancer.Relative expression of 18SRNA and HPRT-1 showed no significant differences.Significant differences in MMP2 and MMP9 were found between cancer and normal tissue,but not between gastritis and normal tissue.Absolute quantification of MMP9 echoed this pattern,but differential expression of MMP2 proved conflictive.Analysis by qGZ indicated significant differences between cancer and normal tissue in MMP-2,total MMP-9,250 and 110 kDa bands.CONCLUSION:MMP9 expression is enhanced in gastric cancer compared to normal mucosa;interpretation of differential expression of MMP2 is difficult to establish.

Identifying the molecular basis of Jinhong tablets against chronic superficial gastritis via chemical profile identification and symptom-guided network pharmacology analysis

Chronic superficial gastritis(CSG)is a common disease of the digestive system that possesses a serious pathogenesis.Jinhong tablet(JHT),a traditional Chinese medicine(TCM)prescription,exerts therapeutic effects against CSG.However,the molecular basis of its therapeutic effect has not been clarified.Herein,we employed ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q/TOF-MS)based chemical profile identification to determine the chemical components in JHT.Further,we applied network pharmacology to illustrate its molecular mechanisms.A total of 96 chemical constituents were identified in JHT,31 of which were confirmed using reference standards.Based on the bioinformatics analysis using the symptom-guided pharmacological networks of“chi,”“blood,”“pain,”and“inflammation,”and target screening through the interaction probabilities between compounds and targets,matrix metalloproteinase 2(MMP2),dopamine d2 receptor(DRD2),and Aldo-keto reductase family 1 member B1(AKR1B1)were identified as key targets in the therapeutic effect exhibited by JHT against CSG.Moreover,according to the inhibitory activities presented in the literature and binding mode analysis,the structural types of alkaloids,flavonoids,organic acids,including chlorogenic acid(10),caffeic acid(13),(-)-corydalmine(33),(-)-isocorypalmine(36),isochlorogenic acid C(38),isochlorogenic acid A(41),quercetin-3-O-a-L-rhamnoside(42),isochlorogenic acid B(47),quercetin(63),and kaempferol(70)tended to show remarkable activities against CSG.Owing to the above findings,we systematically identified the chemical components of JHT and revealed its molecular mechanisms based on the symptoms associated with CSG.

Evaluation of the Gastric Microbiome in Patients with Chronic Superficial Gastritis and Intestinal Metaplasia

Objective To evaluate the gastric microbiome in patients with chronic superficial gastritis(CSG)and intestinal metaplasia(IM)and investigate the influence of Helicobacter pylori(H.pylori)on the gastric microbiome.Methods Gastric mucosa tissue samples were collected from 54 patients with CSG and IM,and the patients were classified into the following four groups based on the state of H.pylori infection and histology:H.pylori-negative CSG(n=24),H.pylori-positive CSG(n=14),H.pylori-negative IM(n=11),and H.pylori-positive IM(n=5).The gastric microbiome was analyzed by 16S rRNA gene sequencing.Results H.pylori strongly influenced the bacterial abundance and diversity regardless of CSG and IM.In H.pylori-positive subjects,the bacterial abundance and diversity were significantly lower than in H.pylori-negative subjects.The H.pylori-negative groups had similar bacterial composition and bacterial abundance.The H.pylori-positive groups also had similar bacterial composition but different bacterial relative abundance.The relative abundance of Neisseria,Streptococcus,Rothia,and Veillonella were richer in the I-HP group than in G-HP group,especially Neisseria(t=175.1,P<0.001).Conclusions The gastric microbial abundance and diversity are lower in H.pylori-infected patients regardless of CSG and IM.Compared to H.pylori-positive CSG group and H.pylori-positive IM,the relative abundance of Neisseria,Streptococcus,Rothia,and Veillonella is higher in H.pylori-positive patients with IM than in H.pylori-positive patients with CSG,especially Neisseria.

Serum protein profiles suggest a possible link between qi deficiency constitution and Pi-qi-deficiency syndrome of chronic superficial gastritis

Objective:To identify potential serum protein candidates involved in linking the traditional Chinese medicine(TCM)-defined qi deficiency constitution(QDC)to Pi-qi-deficiency syndrome(PQDS)of chronic superficial gastritis(CSG).Methods:Using participants with the TCM-defined balanced constitution as a control population,labelfree quantitative proteomics was adopted to identify differentially expressed proteins(DEPs)in serum samples from two case populations:case population 1(participants with QDC)and case population 2(patients with PQDS of CSG).The DEPs discovered in both case populations were analyzed to identify common DEPs as potential candidates for proteins involved in the link between QDC and PQDS.Based on Kyoto Encyclopedia of Genes and Genomes pathway(KEGG)and Gene Ontology(GO)enrichment analysis and analysis of proteineprotein interaction networks,we evaluated the possible functions of these potential serum candidates.Results:We discovered 24 and 28 proteins that were differentially expressed in case populations 1 and 2,respectively,compared with the control population.Hierarchical clustering analysis showed that the expression profile of DEPs of individuals from the same population clustered well,while those from different populations were segregated.Furthermore,GO analysis revealed the 10 DEPs that were common to both case populations to be mainly associated with negative regulation of cellular metabolic and immune system processes while KEGG analysis indicated these proteins to be associated with complement and coagulation cascades and peroxisome proliferator-activated receptor signaling.Notably,serum levels of C4b-binding protein beta chain,glycosylphosphatidylinositol-specific phospholipase D1 and MS-F1 light chain variable region proteins were notably higher in the two case populations compared with the control,particularly in the case of CSG with PQDS.Conclusion:The results presented here provide new insights into the molecular mechanisms underlying development of PQDS of CSG from QDC,and suggest candidate serum biomarkers for future application in integrative medicine.

Bioinformatics Research on Chronic Superficial Gastritis of Pi-deficiency Syndrome by Gene Arrays

Objective： To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis （CSG） with the Pi-deficiency syndrome （PDS） and those of the non- Pi-deficiency syndrome （non-PDS）, i.e. patients of CSG with Pi （脾）-Wei （胃） dampnese-heat syndrome and healthy persons. Methods： With the BRB-Array Tools software package, original data collection and bioinformatic analysis of gene arrays were conducted in 6 CSG patients of PDS （CSG-PDS）, 6 CSG patients of non-PDS （CSG-nPDS）, and 6 healthy volunteers （Normal）. Results： Compared with non-PDS, the gene expressions in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly. Conclusions： The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.

A Long-Term Follow-up Study of Chronic Gastritis with Fibergastroscope

A more than 5 year follow up study about 48 cases of chronic atrophic gastritis(CAG) and 100 cases of chronic superficial gastritis (CSG) is reported in CAG, it is found that 35.5% of the patients were improved, 41.6% stabilized, 12.5% deteriorated, 6.2% recovered, and 4.2% cancerated. In CSG, it is found that 40% of the patients were improved, 48% stabilized, 6% deteriorated, 5% recovered and 1% cancerated. It is concluded that CAG group has a higher rate of cancerization than that of CSG group.

Detection In Situ of E-cadherin,β-catenin,TCF4 and CDX2 in Various Gastric Diseases

Objective： To investigate the expressions of E-cadherin, β-catenin, transcription factor 4 （TCF4）, which are related with Wnt/β-catenin signaling pathway, and Caudal-type Homeobox Protein 2 （CDX2） originated from intestine, in various gastric diseases and its influence on gastric carcinogenesis and differentiation. Methods： A total of 319 various gastric diseases samples from gastroscopy were detected in situ using immunohistochemistry, and data were analyzed with Х^2 test and Kendall＇s tau-b rank correlation coefficient analysis. Results： Along with the progression from superficial gastritis （SG） to atrophic gastritis （AG） to gastric cancer （GC）, the expressions of E-cadherin and β-catenin appeared a decreasing tendency in total, while β-catenin emerged a decreasing tendency in the cell membrane and an increasing in the nucleus, and the highly positive expression rate of intranuclear transcription factor TCF4 appeared an increasing tendency. The transcription factor CDX2 had a higher expression in AG and GC than that in SG. The expression rates of the four proteins above in intestinal type adenocarcinoma were all higher than those in diffuse type carcinoma. There were positive correlations between E-cadherin and β-catenin expressed in cytoplasm, between β-catenin expressed in cell membrane and β-catenin in cytoplasm, between β-catenin expressed in cytoplasm and β-catenin in nucleus, between β-catenin and TCF4 in nucleus, between TCF4 and CDX2 in nucleus, respectively （P〈0.05）. And β-catenin and CDX2 had no relevance in nucleus （P〉0.05）. Conclusion： We found that the four proteins were involved in gastric carcinogenesis and the differentiation to intestinal type adenocarcinoma.