Helicobacter Pylori In Patients With Bile Pigment Calculus In Biliary System

Objective:This experiment explored the relationship between Hp infection and the formation of pigmented calculus and the possible mechanism of gallstone formation through the study on Hp in the center of gallstones , bile and bile duct mucosa.Methods:Regular, anaerobic and Helicobacter pylori cultures were performed for all bile (include 35 samples of bile pigment calculus and 10 samples of the controls). Helicobacter pylori DNA in bile, bile duct mucosa and pigmented calculus were detected by PCR , and Western- blot was performed to detect Helicobacter pylori associated protein in all bile samples. The bile duct mucosa was stained by Warthin-Starry.Results:1.The regular, anaerobic and Helicobacter pylori cultures were found 15 positive of regular and anaerobic in 35 bile samples of bile pigment calculus and negative of regular in the controls. Helicobacter pylori cultures were found no bacteria growing in all bile samples.2.In 35 samples of bile pigment calculus ,the positive rate of Hp -DNA PCR amplification in the center of gallstones , bile and bile duct mucosa was 14.29% , 31.43 % and 56.67 % respectively. The Hp-DNA in the bile and bile duct mucosa of control group were negative. There were significant differences between the positive rates of Hp -DNA PCR amplification in bile(p<0.01) and bile duct mucosa(p<0.05) of experimental group and those of control group. In experimental group, there were significant differences between the positive rate of Hp -DNA PCR amplification in bile duct mucosa and those in bile(p<0.05) and center of gallstones(p<0.01), and the Hp–DNA in bile and bile duct mucosa of all the samples of which Hp–DNA in center of gallstones were postive were postive.3.PCR showed positive in 11 of 35 bile samples,of which 7contained anti-CagA antibodies,6 contained Vac A. Vacuolating cytotoxin associated 35kD and 30 kD glycoprotein ,Ure B and UreA were also found in the bile. PCR showed negative in 23 of 35 bile samples ,of which 1 contained anti-CagA antibodies, Ure B and UreA found in the bile .The associated protein of Helicobacter pylori infection was not found in the controls4.Bacteria resembling Helicobacter pylori were found in 7(23.33 %) of 30 bile duct mucosa samples stained by Warthin-Starry and Hp–DNA of these samples was positive, however the results in the control group were negative.Conclusions :The presence of Helicobacter pylori DNA in biliary tract system and associated protein of Helicobacter pylori infection in some bile samples may be represent risk factors for the formation of pigmented calculus...