Systemic analysis of urinary stones from the Northern, Eastern, Central, Southern and Southwest China by a multi-center study.

BACKGROUND: To provide some basis for the prevention of urinary stones in general population, we did a systemic analysis of urinary stones from Northern, Eastern, Central, Southern and Southwest China by a multi-center study. METHODS: A total of 11,157 urinary stones from Northern, Eastern, Central, Southern and Southwest China were obtained and analyzed by Fourier transform infrared spectroscopy. Combined with scanning electron microscopy and X-ray energy spectrometer, urinary stones were classified into different types. Furthermore, the correlation between stone types and clinical characteristics, as well as their regional distribution were elucidated. RESULTS: Calcium oxalate stones were the most common type in each region, followed by calcium oxalate-calcium phosphate mixed stones, uric acid stones and calcium phosphate stones. The distribution of calcium oxalate stones were highest prevalence in Southwest China (67.9%, P < 0.05), followed by Eastern and Northern China. Anhydrous uric acid stones, with a constituent ratio of 19.3% in Southern China, and 13.7% in Central China, were significantly higher than that in other regions (P < 0.05). Elements analysis indicated varieties among stone types as well as distribution regions. Moreover, the clinical characteristics were highly correlated with stone types and anatomical locations but not their distribution regions. CONCLUSIONS: The material and elements composition of urinary stones among different regions showed some varieties. Calcium oxalate stone has the highest constituent ratio in Southwest China, while anhydrous uric acid stone has the highest constituent ratio in Southern China. Moreover, the clinical characteristics were highly correlated with stone types and anatomical locations but not their distribution regions.

The systematic classification of urinary stones combine-using FTIR and SEM-EDAX.

BACKGROUND: To explore underlying mechanism of urinary stones formation, the composition and microstructure of urinary stones were analyzed systematically with a large sample study from China. MATERIALS AND METHODS: A total of 2437 urinary stones were obtained from the urology department at our Hospital. The composition of the stones was analyzed by Fourier transform infrared spectroscopy (FTIR). Meanwhile, the microstructure and element distribution were observed with scanning electron microscopy combined with element distribution analysis (SEM-EDAX). RESULTS: Urinary stones were classified into eight types, that were consisted of calcium oxalate stones (1301/2437, 53.39%), calcium phosphate stones (131/2437, 5.38%), anhydrous uric acid stones (434/2437, 17.81%), magnesium ammonium phosphate stones (12/2437, 0.49%), sodium urate stones (5/2437, 0.21%), brushite stones (4/2437, 0.16%), cystine stones (3/2437, 0.12%) and mixed stones (547/2437, 22.45%, ten subtypes were included). Under SEM, they displayed distinct microstructures: plank-like, brick-like, polyhedron or paliform crystals for calcium oxalate stones, similar sized echin-sphere or rough bulbiform or tree bark-like crystals for calcium phosphate stones, rotten-wood-like or petrous crystals for anhydrous uric acid stones, globular or gallet-like crystals for magnesium ammonium phosphate stones, sawdust-like crystals for sodium urate stones, broken-wood-like crystals for brushite stones, stacking hexagonal cystine crystals for cystine stones, and two or more of the above crystals for mixed stones. Meanwhile, they also presented distinct elemental composition and distribution by EDAX. CONCLUSIONS: Urinary stones can be classified into eight types, and exhibit a diversity of microstructure and elemental compositions in China. The formation process of different types of urinary stones may be diverse.

A comparative study of mud-like and coralliform calcium carbonate gallbladder stones.

To gain insight to underlying mechanism of the formation of calcium carbonate (CaCO3 ) gallbladder stones, we did comparative study of stones with mud appearance and those with coralliform appearance. A total of 93 gallbladder stones with mud appearance and 50 stones with coralliform appearance were analyzed. The appearance, color, texture, and the detection of Clonorchis sinensis eggs by microscopic examination were compared between the two groups. Then, the material compositions of stones were analyzed using Fourier Transform Infrared spectroscopy and the spectrogram characteristics were compared. Moreover, microstructure characteristics of the two kinds of stones were observed and compared with Scanning Electron Microscopy. Mud-like gallbladder stones were mainly earthy yellow or brown with brittle or soft texture, while coralliform stones were mainly black with extremely hard texture, the differences between the two groups was significant (p < .05). The analytic results of FTIR spectroscopy showed that 95.7% (89/93) of the mud-like gallbladder stones were CaCO3 stones, and mainly aragonite; while all of the coralliform stones were CaCO3 stones, and mainly calcite (p < .05). Meanwhile, microscopic examination indicated that the detection rate of Clonorchis sinensis eggs in mud-like CaCO3 stones was lower than that in coralliform CaCO3 stones (p < .05), and that in aragonite CaCO3 stones was lower than that in calcite CaCO3 stones(p < .05). Mud-like CaCO3 stones mainly happened to patients with cystic duct obstruction. Clonorchis sinensis infection was mainly associated with coralliform (calcite) CaCO3 stones. Cystic duct obstruction was mainly associated with mud-like (aragonite) CaCO3 stones.

[Gallbladder Stone Types in Patients with Cholelithiasis and Clonorchis sinensis Infection in the Endemic Area of Clonorchiasis].

OBJECTIVE: To investigate the relationship between Clonorchis sinensis infection and the gallbladder stone type in patients with cholelithiasis in the endemic area of clonorchiasis. METHODS: Gallbladder stones were collected from 598 patients with cholelithiasis through minimally invasive gallbladder-preserving cholelithotomy in the Sixth People's Hospital of Nansha District from May 2009 to October 2012. The stone samples were analyzed for composition by Fourier transform infrared spectroscopy to identify their types. The Clonorchis sinensis eggs were detected in the stones by microscopic examination, and the detection rates of eggs were calculated for different stone types. Then the clinical characteristics and biochemical indicators were compared among patients with different types of stones, as well as between Clonorchis sinensis egg-positive and -negative patients with the calcium-carbonate type of stones. Some calcium-carbonate stones positive for Clonorchis sinensis eggs were randomly selected for further scanning electron microscopy (SEM) examination. RESULTS: Of the stones from 598 patients, 234 (39.1%) were cholesterol type, 133 (22.2%) bile pigment type, 112(18.7%) calcium-carbonate type, 86 (14.4%) mixed types and 33(5.5%) were others. The detection rate of Clonorchis sinensis eggs in these types was 6% (15/234), 44% (59/133), 60% (67/112), 36% (31/ 86) and 30% (10/33), respectively, being highest in calcium-carbonate stones while lowest in cholesterol stones. The Co2-combining power of the plasma was higher in patients with calcium-carbonate and mixed stones than in those with cholesterol stones (P < 0.05), and the CO2-combining power of the bile and biliary pH were both higher in patients with calcium-carbonate types than in those with other types (P < 0.05). In addition, in patients with calcium-carbonate stones, the CO2-combining powers of the plasma and the bile, as well as biliary pH were all higher in the egg-positives than in the egg-negatives. Further, both light microscopy and SEM revealed adherence of the Clonorchis sinensis eggs to calcium-carbonate crystals. CONCLUSION: The infection rate of Clonorchis sinensis is higher in patients with calcium-carbonate gallbladder stones than in those with other types of stones.

Clonorcis sinensis eggs are associated with calcium carbonate gallbladder stones.

Calcium carbonate gallbladder stones were easily neglected because they were previously reported as a rare stone type in adults. The aim of this study was to investigate the relationship between calcium carbonate stones and Clonorchis sinensis infection. A total of 598 gallbladder stones were studied. The stone types were identified by FTIR spectroscopy. The C. sinensis eggs and DNA were detected by microscopic examination and real-time fluorescent PCR respectively. And then, some egg-positive stones were randomly selected for further SEM examination. Corresponding clinical characteristics of patients with different types of stones were also statistically analyzed. The detection rate of C. sinensis eggs in calcium carbonate stone, pigment stone, mixed stone and cholesterol stone types, as well as other stone types was 60%, 44%, 36%, 6% and 30%, respectively, which was highest in calcium carbonate stone yet lowest in cholesterol stone. A total of 182 stones were egg-positive, 67 (37%) of which were calcium carbonate stones. The C. sinensis eggs were found adherent to calcium carbonate crystals by both light microscopy and scanning electron microscopy. Patients with calcium carbonate stones were mainly male between the ages of 30 and 60, the CO2 combining power of patients with calcium carbonate stones were higher than those with cholesterol stones. Calcium carbonate gallbladder stones are not rare, the formation of which may be associated with C. sinensis infection.

The systematic classification of gallbladder stones.

BACKGROUND: To develop a method for systematic classification of gallbladder stones, analyze the clinical characteristics of each type of stone and provide a theoretical basis for the study of the formation mechanism of different types of gallbladder stones. METHODOLOGY: A total of 807 consecutive patients with gallbladder stones were enrolled and their gallstones were studied. The material composition of gallbladder stones was analyzed using Fourier Transform Infrared spectroscopy and the distribution and microstructure of material components was observed with Scanning Electron Microscopy. The composition and distribution of elements were analyzed by an X-ray energy spectrometer. Gallbladder stones were classified accordingly, and then, gender, age, medical history and BMI of patients with each type of stone were analyzed. PRINCIPAL FINDINGS: Gallbladder stones were classified into 8 types and more than ten subtypes, including cholesterol stones (297), pigment stones (217), calcium carbonate stones (139), phosphate stones (12), calcium stearate stones (9), protein stones (3), cystine stones (1) and mixed stones (129). Mixed stones were those stones with two or more than two kinds of material components and the content of each component was similar. A total of 11 subtypes of mixed stones were found in this study. Patients with cholesterol stones were mainly female between the ages of 30 and 50, with higher BMI and shorter medical history than patients with pigment stones (P<0.05), however, patients with pigment, calcium carbonate, phosphate stones were mainly male between the ages of 40 and 60. CONCLUSION: The systematic classification of gallbladder stones indicates that different types of stones have different characteristics in terms of the microstructure, elemental composition and distribution, providing an important basis for the mechanistic study of gallbladder stones.

Microscopic examination of gallbladder stones improves rate of detection of Clonorchis sinensis infection.

To improve the rate of detection of Clonorchis sinensis infection, we compared different specimens from patients with cholecystolithiasis. Feces, gallbladder bile, and gallbladder stones collected from 179 consecutive patients with cholecystolithiasis underwent microscopic examination, and according to the results, 30 egg-positive and 30 egg-negative fecal, gallbladder bile, and gallbladder stone specimens, respectively, underwent real-time fluorescent PCR. The detection rates of eggs in feces, bile, and gallbladder stones were 30.7%, 44.7%, and 69.8%, respectively, and the differences were statistically significant (P<0.01). The PCR results confirmed that the eggs in the specimens were C. sinensis eggs. Eggs in the feces were "fresh" and in the gallbladder stones were "old." Microscopic examination of gallbladder stones may improve the detection rates of C. sinensis infection, which is important for developing individualized treatments to prevent the recurrence of gallbladder stones and to prevent the occurrence of severe liver damage and cholangiocarcinoma.

A microstructural study of gallbladder stones using scanning electron microscopy.

OBJECTIVE: The objective of this study was to describe the microstructure of different types of gallbladder stones to better understand the basis of gallbladder stone formation. METHODS: Gallbladder stones from 387 patients with cholecystolithiasis were first analyzed by Fourier transform Infrared spectroscopy to identify the type of the gallbladder stone, and they were then examined using scanning electron microscopy to define their microstructure. RESULTS: Cholesterol stones were mainly composed of plate-like or lamellar cholesterol crystals stacked tightly in a radial, cord-like, or irregular staggered arrangement. A small number of bilirubinate particles were seen occasionally. Pigment stones were mainly composed of loosely arranged bilirubinate particles with different shapes (sphere-like, clumping-like, or amorphous). Calcium carbonate stones were composed of calcium carbonate crystals having many shapes (bulbiform, ellipsoid, fagot-shaped, fusiform, hawthorn-shaped, cuboid, button-shaped, lamellar, broken firewood-shaped, rod-shaped, acicular, or crushed sugar cane-shaped). Bulbiform shaped crystals were the most common. Phosphate stones were mainly composed of different sized echin-sphere-like or rough bulbiform crystals. Bilirubinate particles were always adherent. Calcium stearate stones usually had a network structure with adherent bilirubinate particles. Protein stones usually had a honeycomb or chrysanthemum petal-like structure. Cystine stones were composed of hexagonal cystine crystals, some of which had prominent edges. Mixed stones presented different shapes according to their mixed components. CONCLUSION: Different types of gallbladder stones had characteristic shapes and elements. This study provides an objective basis for further research regarding gallbladder stone formation.

[Survey on Clonorchis sinensis infection and bile component of gallstone patients from the Pearl River Delta region in Guangdong Province].

OBJECTIVE: To evaluate the prevalence of Clonorchis sinensis and bile components of gallstone patients from the Pearl River Delta region in Guangdong Province. METHODS: From June 2011 to November 2012, 406 gallstone patients from the Pearl River Delta region in Guangdong Province were enrolled in the study. Fecal specimens, bile and gallbladder stone samples were examined for the presence of C. sinensis eggs by the Kato-Katz method, centrifuged sediment smear microscopy, and triturated stone microscopy, respectively. The blood samples were detected for the antibody against C. sinensis by using colloidal gold immunochromatographic assay. Total calcium, ionized calcium, HCO3-, magnesium, pH, total bilirubin (TBIL), total bile acid (TBA), total cholesterol (CHO), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were analyzed quantitatively in bile of gall bladder. RESULTS: The overall C. sinensis infection rate was 52.2% (212/406). The proportion of males infected with C. sinensis was 64.7% (130/201), higher than that of females (40.0%, 82/205). The content of TBIL [(1 458.0 +/- 681.0) micromol/L], CHO [(4.1 +/- 1.3) mmol/L] and magnesium [(4.8 +/- 1.8) mmol/L] in the bile from the patients under 30 years old, total calcium [(3.0 +/- 1.3) mmol/L], ionized calcium [(1.5 +/- 0.7) mmol/L], TBA [(114.6 +/- 54.5) mmol/L], CHO [(5.1 +/- 1.7) mmol/L], TBIL [(1 396.0 +/- 615.0) micromol/L], GGT [(1 562.0 +/- 583.0) U/L] and ALP [(263.0 +/- 94.0) U/L] in 31-40 years-old, or CHO [(5.4 +/- 2.2) mmol/L] in 41-50 years-old in egg-positive group was lower than that of the corresponding negative groups [(2 759.0 +/- 969.0) micromol/L, (7.5 +/- 2.5), (7.5 +/- 2.2), (3.8 +/- 1.6), (1.9 +/- 1.0), (144.1 +/- 63.4), and (9.9 +/- 2.5) mmol/L, (1 892.0 +/- 584.0) micromol/L, (2457.0 +/- 988.0) and (535.0 +/- 196.0) U/L, and (7.9 +/- 2.3) mmol/L], respectively (P < 0.05). While, the content of HCO3- [(22.7 +/- 5.1) mmol/L], pH (7.6 +/- 0.4) in 31-40 years-old group and pH (7.6 +/- 0.4) in the patients aged above 50 years-old was higher than that of the negatives [(17.3 +/- 6.9) mmol/L, 7.4 +/- 0.2, and 7.5 +/- 0.3] (P < 0.05). CONCLUSION: In the Pearl River Delta region in Guangdong Province, C. sinensis prevalence is high in the gallstone patients. There are differences in bile components between C. sinensis-infected and non-infected patients.

[Detection of Clonorchis sinensis eggs in the ground gallbladder stones by microscopy].

Sera, feces, bile and gallbladder stones were collected from 179 patients who accepted gallbladder-preserving cholelithotomy during the period of January to June 2010 at the general surgery department in the Second People's Hospital of Panyu District in Guangzhou. Rapid colloidal gold immunochromatography was used to detect IgG against Clonorchis sinensis. C. sinensis eggs were examined by fecal direct smear, and in bile sediments and ground gallbladder stones. The results showed that the positive rate of rapid colloidal gold immunochromatographic assay for IgG was 51.4%, and the egg positive rate in feces, bile sediments and gallbladder stones was 30.7%, 44.7% and 69.8%, respectively. The detection rate of fecal direct smear was the lowest, while that of the gallbladder stone examination was the highest (P < 0.05). Those patients with egg-positive feces and/or bile sediments were all with egg-positive gallbladder stones.

Cholecystolithiasis is associated with Clonorchis sinensis infection.

BACKGROUND: The objective of this study was to analyze gallbladder stones for direct evidence of a relationship between Clonorchis sinensis infection and gallbladder stones formation. METHODOLOGY: We 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.

Development of a real-time PCR assay for the detection of Clonorchis sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis.

High prevalence of cholecystolithiasis in parts of East Asia has been postulated to be associated with Clonorchis sinensis infection. This study describes the development of a TaqMan-based real-time PCR assay for the detection of C. sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis. Primers and probe targeting the cytochrome c oxidase subunit 1 gene of mitochondrial DNA proved to be highly specific for C. sinensis and did not amplify other related heterogeneous DNA samples. The detection limit of this assay was 0.1 pg of adult C. sinensis genomic DNA. All of the egg-positive samples determined by microscopy yielded positive results by real-time PCR assay and that genetic testing of gallbladder stones using real-time PCR was considered as the most effective means for assessing C. sinensis infection status. This assay not only contributes to a greater understanding of stone pathogenesis but also benefits patients with cholecystolithiasis by facilitating effective diagnosis, treatment, and relapse prevention.

Tiny cystine stones in the gallbladder of a patient with cholecystolithiasis complicating acute cholecystitis: a case report.

Cystine stones, the main component of which is cystine, are very common urinary calculi, but are rare in the gall bladder. In animals, there has been only one report of cystine gallstones in tree shrews, and to our knowledge, this is the first report of cystine gallstones in humans.