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目的探讨幽门螺杆菌(Hp)感染与胆色素结石形成的关系。方法取35例胆色素性结石患者(实验组)及10例对照组患者胆汁进行需氧菌、厌氧菌及Hp培养,采用PCR方法检测胆囊管黏膜、胆管黏膜、胆汁和胆石核心中的Hp-DNA;所有黏膜用Warthin-Starry银染色法染色,Western印迹法检测胆汁中Hp感染相关蛋白。结果所有胆汁培养均未发现Hp生长。实验组胆石核心、胆汁和黏膜的Hp-DNA扩增阳性率分别为14.29%、31.43%和56.67%。Hp-DNA阳性胆汁中,检出细胞毒素相关抗原(Cag A)7例,细胞空泡毒紊(Vae A)6例,同时检出Hp Vac A相关亚单位(相对分子质量37000)、糖蛋白(相对分子质量35000)、尿素酶B和尿素酶A。实验组30例胆管黏膜经Warthin- Starry染色后有7例(23.33%)镜下观察到类似Hp的菌体。对照组胆汁和黏膜Hp-DNA阴性,胆汁中未检出Hp感染相关蛋白.黏膜Warthin-Starry染色后,镜下未观察到类似Hp的菌体。结论胆色素结石患者胆道系统中存在Hp感染依据.并存在多种Hp感染相关蛋白,Hp感染可能参与胆色素性结石的形成。
Objective To investigate the relationship between Helicobacter pylori (Hp) infection and the formation of bile pigment stones. Methods Thirty-five patients with bile duct stones (experimental group) and 10 control subjects were enrolled in this study. Aerobic, anaerobic and Hp cultures of bile were performed. The expression of Hp in mucosa, bile duct, bile and gallstone -DNA. All mucosa were stained with Warthin-Starry silver staining and Western blotting was used to detect Hp infection-related proteins in bile. Results No Hp growth was found in all bile cultures. The positive rates of Hp-DNA amplification in gallstone core, bile and mucosa in experimental group were 14.29%, 31.43% and 56.67% respectively. In Hp-DNA positive bile samples, 7 cases of cytotoxic antigen (Cag A) and 6 cases of Vae A were detected. Hp Vac A related subunit (relative molecular mass 37000), glycoprotein (Relative molecular weight 35000), urease B and urease A In the experimental group, 7 cases (23.33%) of the bile duct mucosa stained with Warthin-Starry showed Hp-like cells. Hp-DNA was negative in the bile and mucosa of the control group, and no Hp-related protein was detected in the bile.After the Warthin-Starry staining of the mucosa, no Hp-like cells were observed under the microscope. Conclusions Hp infection exists in the biliary system of patients with biliary pigment stones, and there are many proteins related to Hp infection. Hp infection may be involved in the formation of biliary pigment stones.