Abstract
BackgroundThe objective of this study was to analyze gallbladder stones for direct evidence of a relationship between Clonorchis sinensis infection and gallbladder stones formation.MethodologyWe investigated one hundred eighty-three gallbladder stones for the presence of Clonorchis sinensis eggs using microscopy, and analyzed their composition using Fourier transform infrared spectroscopy. We confirmed the presence of Clonorchis sinensis eggs in the gallbladder stones using real-time fluorescent PCR and scanning electron microscopy.Principal Findings Clonorchis sinensis eggs were detected in 122 of 183 gallbladder stones based on morphologic characteristics and results from real-time fluorescent PCR. The proportion of pigment stones, cholesterol stones and mixed gallstones in the egg-positive stones was 79.5% (97/122), 3.3% (4/122) and 17.2% (21/122), respectively, while 29.5% (18/61), 31.1% (19/61) and 39.3% (24/61) in the egg-negative stones. The proportion of pigment stone in the Clonorchis sinensis egg-positive stones was higher than in egg-negative stones (P<0.0001). In the 30 egg-positive stones examined by scanning electron microscopy, dozens or even hundreds of Clonorchis sinensis eggs were visible (×400) showing a distinct morphology. Many eggs were wrapped with surrounding particles, and in some, muskmelon wrinkles was seen on the surface of the eggs. Also visible were pieces of texture shed from some of the eggs. Some eggs were depressed or without operculum while most eggs were adhered to or wrapped with amorphous particles or mucoid matter (×3000).Conclusion Clonorchis sinensis eggs were detected in the gallbladder stones which suggests an association between Clonorchis sinensis infection and gallbladder stones formation, especially pigment stones.
Highlights
Cholecystolithiasis or the presence of gallstones in the gallbladder is a common disease with a mean prevalence rate of 10% based on epidemiological studies [1]
Clonorchis sinensis eggs were detected in the gallbladder stones which suggests an association between Clonorchis sinensis infection and gallbladder stones formation, especially pigment stones
C. sinensis eggs are yellow-brown in color, resemble sesame seeds, have a thick shell and a small operculum in the front end, shouldering in the junction of the shell and operculum, with occasional visible small protrusions in the obtuse posterior end
Summary
Cholecystolithiasis or the presence of gallstones in the gallbladder is a common disease with a mean prevalence rate of 10% based on epidemiological studies [1]. The incidence rate increases with changes in dietary patterns and with the development of an aging population [2,3,4,5,6]. Clonorchiasis, known as liver fluke disease, is an important amphixenosis. It is mainly distributed in East Asia and Southeast Asia, including China, DPR.Korea, the Republic of Korea, Vietnam, and the Philippines [13]. Based on the second national survey on parasitic diseases between 2001 and 2004 in China, the overall C. sinensis infection rate of the surveyed population was 0.58% [14,15]. The objective of this study was to analyze gallbladder stones for direct evidence of a relationship between Clonorchis sinensis infection and gallbladder stones formation
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