The Preliminary Research On The Relationship Of Clonorchis Sinensis Infection And Gallbladder Stones From Patients With Cholecystolithiasis

Gallstones is common diseases in China and the world, epidemiologic studies indicate mean prevalence rates of about10%in population.The incidence of gallbladder stones shows an ascendant trend accompanies with the improvement of people’s living standard and the change of dietary patterns as well as the development of an aging population. In order to find out the causative factor of gallbladder stones, many scholars have done lots of relevant research, but there are still many phenomenon still can’t be explained.Many researchers in China and abroad have done some research on the relationshiop of Clonorchis sinensis infection and gallstones and concluded that Clonorchis sinensis infection involved in the formation of bile duct stones rather than gallbladder stones,but we detected C. sinensis eggs in gallbladder stones through light microscope incidentally, then a penetrating study was performed.Objective:This study was to approach the relationship between Clonorchis sinensis infection and gallbladder stones of patients with cholecystolithiasis.Methods:(1) Composition analysis of gallbladder stones by Fourier transform infrared (FTIR) spectroscopy:through comparisons of the spectra between gallbladder stones and standard controls, gallbladder stones were grouped as cholesterol stones, bile pigment stones and mixed stones.(2) Gallbladder stones were ground and smeared with0.9%NaCl, then, the filtrate was smeared onto labeled slides and observed with an Olympus System Microscope (BX51, Japan). Each filtrate was smeared onto3to4slides.(3) Detection of C. sinensis DNA in the gallbladder stones by real-time fluorescent PCR.(4) Gallbladder stones were taken and then fixed on the sample table and dried at60℃overnight. Then, the stones were sputter-coated with gold (ETD-2000, China) and observed under a ZEISS (EVO LS10, Germany) SEM and photographed. Meanwhile, the elements composition was analyzed with X-ray energy spectrometer.Results:(1) The main composition of gallbladder stones were bilirubin and bilirubinate, cholesterol, calcium carbonate and calcium stearate. According to the main composition, gallbladder stones were grouped as cholesterol stones, bile pigment stones and mixed stones.(2) C. sinensis eggs were detected in the gallbladder stones, and were preliminarily judged as C. sinensis eggs morphologically, which were further confirmed by real-time fluorescent PCR.(3) The C. sinensis egg-positive gallbladder stones were mainly pigment stones. Gallbladder stones from patients in the epidemic area of clonorchiasis sinensis were mainly pigment stones, while gallbladder stones from patients in the non-epidemic area of clonorchiasis sinensis were mainly cholesterol stones.(4) The scanning electron microscopy and X-ray spectroscopy analysis showed that microstructure and elemental composition of all types of stone were different and had different characteristics, the elemental composition of egg-positive stones was particularly complicated.Conclusion:The research showed that C. sinensis eggs were detected in the gallbladder stones, and the C. sinensis egg-positive gallbladder stones were mainly pigment stones, indicating that C. sinensis infection are associated with gallbladder stones especially pigment stones. The microstructure and elemental composition of all types of stone were different and had different characteristics; the elemental composition of egg-positive stones was particularly complicated.