AIMTo establish a LC-MS(n) method for the identification of anisodamine and its metabolites in rat feces.

METHODSFeces samples were collected after single administration of 25 mg x kg(-1) anisodamine to rats, and dipped in water for 1 h. Samples were then extracted by ethyl acetate. The pretreated samples were separated on a reversed-phase C18 column using a mobile phase of methanol / 0.01% triethylamine (adjusted to pH 3.5 with formic acid) (60 : 40, v/v) and detected by LC-MS". Identification of the metabolites and elucidation of their structures were performed by comparing their changes in molecular masses (deltaM), retention-times and full scan MS(n) spectra with those of the parent drug and blank feces.

RESULTSThe parent drug and its seven metabolites (6beta-hydroxytropine, nor-6beta-hydroxytropine, aponoranisodamine, apoanisodamine, noranisodamine and hydroxyanisodamine, tropic acid) were found in rat feces.

CONCLUSIONThis method is sensitive, rapid, simple, effective, and suitable for the rapid identification of drug and its metabolites in biologic samples.

Animals ; Feces ; chemistry ; Rats ; Rats, Wistar ; Solanaceous Alkaloids ; analysis ; metabolism ; Tandem Mass Spectrometry ; methods

No abstract available.
Antigens, Bacterial/*analysis ; Feces/*microbiology ; Helicobacter Infections/*diagnosis ; Helicobacter pylori/*isolation & purification ; Humans ; Immunoenzyme Techniques

OBJECTIVETo explore the possibility of Bacteroides spp. as fecal contamination indicator bacteria with real-time quantitative PCR (RT-PCR) assay through analyzing the correlation between Bacteroides spp. and coliform group in external environment.

METHODSQuantity of coliform group and Bacteroides in water samples were detected by most-probable-number method (MPN) and RT-PCR, respectively, and their detection correlation was evaluated with linear correlation analysis. Both methods were also applied to detect the contaminated time limits and river water samples collected at four sampling sites in three different times.

RESULTSSeventy two hours were needed for the numeration of coliform group with MPN method, while RT-PCR could detect Bacteroides within 3 hours. The contaminated time limit of indoor and outdoor water samples of coliform group was more than 40 days and 9 days, and Bacteroides 13 days and 5 days, respectively. Also, the positive correlation between the quantity of Bacteroides and coliform group in outdoor water samples was obtained, the quantity of Bacteroides was from 8.3 × 10(6) copies/ml to less than 10(4) copies/ml during the first day to the fifth day, while coliform group was 4.3 × 10(6) MPN/100 ml to 2.4 × 10(3) MPN/100 ml. A 100% coincidence rate of the detection results with both methods was also observed. These results indicated that the detection results of both methods had perfect consistency.

CONCLUSIONBacteroides spp. can be potentially used as fecal contamination indicator bacteria with RT-PCR rapid detection.

Bacteroides ; Environmental Microbiology ; Environmental Monitoring ; methods ; Escherichia coli ; Feces ; microbiology ; Rivers ; microbiology ; Water Pollutants ; analysis

OBJECTIVETo compare the sensitivity, the specificity and the anti-jamming of several excrement occult blood experimental techniques. To evaluate the effect of transferrin (Tf) in the excrement in the digestive tract hemorrhage detection rate.

METHODSFor 600 patients of clinical suspicious digestive tract hemorrhage, take their excrement specimen, using the chemical process (pyramidon semi-quantitative examination law) to detect hemoglobin (Hb), and using monoclonal antibody colloidal gold method to detect Hb and Tf.

RESULTSFinally the hemoglobin chemical process (hereafter refers to as chemical process) to detect upper gastrointestinal hemorrhage with the positive rate 57.3%, and the detection of hemorrhage of lower digestive tract's positive rate is 44.8%; Hemoglobin monoclonal antibody colloidal gold method (hereafter refers to as colloid gold law) to examine upper gastrointestinal hemorrhage with a positive rate 60.4%, under examination hemorrhage with positive rate 77.6%; transferrin monoclonal antibody colloidal gold method (hereafter refer to as transferrin law) to examine upper gastrointestinal hemorrhage with a positive rate 82.3%, examination hemorrhage of lower digestive tract with a positive rate 66.4%; The union examination law (hemoglobin and transferrin to be detected twice, once positive that is positive) examines upper gastrointestinal hemorrhage the positive rate is 90.8%, hemorrhage of lower digestive tract's positive rate is 97.6%.

CONCLUSIONExcrement transferrin has the high detection rate in the upper gastrointestinal hemorrhage; Hb and the Tf combined examination may obviously raise the digestive tract hemorrhagic disease's positive detection rate.

Feces ; chemistry ; Gastrointestinal Hemorrhage ; diagnosis ; Gold Colloid ; Humans ; Occult Blood ; Transferrin ; analysis

OBJECTIVETo systemically study the metabolism of ginsenoside Re in rats.

METHODSix SD rats were used divided into 3 groups. After oral administration of ginsenoside Re at a dosage of 100 mg x kg(-1), feces were collected at 12, 24, 36, 48, and 60 h. The microbial transformation metabolites of ginsenoside Re were used as standard references. HPLC-ESI-MS/MS analysis was used in the metabolism studies.

RESULTSix metabolites of ginsenoside Re were detected in rat feces by HPLC-ESI-MS/MS analysis. Their structures were identified as 20(S)-ginsenoside Rg2, 20(S)-ginsenoside Rh1, 20(R)-ginsenoside Rh1, ginsenoside F1, 3-oxo-ginsenoside Rh1 and protopanaxatriol.

CONCLUSIONGinsenoside Re has the same metabolic pathway with microorganisms, to some extent, justified the use of microbial models for mammalian metabolism studies.

Administration, Oral ; Animals ; Biotransformation ; Feces ; chemistry ; Ginsenosides ; analysis ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Fecal immunochemical tests (FITs) are the most widely used non-invasive tests in colorectal cancer (CRC) screening. However, evidence about the direct comparison of the test performance of the self-administered qualitative a laboratory-based quantitative FITs in a CRC screening setting is sparse. METHODS: Based on a CRC screening trial (TARGET-C), we included 3144 pre-colonoscopy fecal samples, including 24 CRCs, 230 advanced adenomas, 622 non-advanced adenomas, and 2268 participants without significant findings at colonoscopy. Three self-administered qualitative FITs (Pupu tube) with positivity thresholds of 8.0, 14.4, or 20.8 μg hemoglobin (Hb)/g preset by the manufacturer and one laboratory-based quantitative FIT (OC-Sensor) with a positivity threshold of 20 μg Hb/g recommended by the manufacturer were tested by trained staff in the central laboratory. The diagnostic performance of the FITs for detecting colorectal neoplasms was compared in the different scenarios using the preset and adjusted thresholds (for the quantitative FIT). RESULTS: At the thresholds preset by the manufacturers, apart from the qualitative FIT-3, significantly higher sensitivities for detecting advanced adenoma were observed for the qualitative FIT-1 (33.9% [95% CI: 28.7-39.4%]) and qualitative FIT-2 (22.2% [95% CI: 17.7-27.2%]) compared to the quantitative FIT (11.7% [95% CI: 8.4-15.8%]), while at a cost of significantly lower specificities. However, such difference was not observed for detecting CRC. For scenarios of adjusting the positivity thresholds of the quantitative FIT to yield comparable specificity or comparable positivity rate to the three qualitative FITs accordingly, there were no significant differences in terms of sensitivity, specificity, positive/negative predictive values and positive/negative likelihood ratios for detecting CRC or advanced adenoma between the two types of FITs, which was further evidenced in ROC analysis. CONCLUSIONS Although the self-administered qualitative and the laboratory-based quantitative FITs had varied test performance at the positivity thresholds preset by the manufacturer, such heterogeneity could be overcome by adjusting thresholds to yield comparable specificities or positivity rates. Future CRC screening programs should select appropriate types of FITs and define the thresholds based on the targeted specificities and manageable positivity rates.
Colonoscopy ; Colorectal Neoplasms/diagnosis* ; Early Detection of Cancer ; Feces ; Hemoglobins/analysis* ; Humans ; Laboratories ; Occult Blood ; Sensitivity and Specificity

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.
Forensic Medicine/methods* ; Body Fluids/chemistry* ; RNA/analysis* ; Feces ; Forensic Genetics ; Semen/chemistry* ; Saliva/chemistry*

OBJECTIVETo investigate the changes and clinical significance of biomarker fecal bile acids (BA) in children with Henoch-Schönlein purpura (HSP).

METHODSNineteen children with HSP and twenty-seven healthy children were enrolled in this study. The stool samples were obtained at the acute and remission phases. Fecal BA levels were measured by high performance liquid chromatography mass spectrometry (HPLC-MS).

RESULTSThe fecal cholic acid level in the HSP remission group was significantly higher than in the HSP acute group and the healthy control group (P<0.016). The fecal chenodeoxycholic acid level in the HSP remission group was significantly higher than in the healthy control group (P<0.016). The levels of fecal secondary colonic bile acids, deoxycholic acid and lithocholic acid, in the HSP acute and remission groups were significantly lower than in the healthy control group(P<0.05, P<0.016 respectively). No significant differences were found in the levels of fecal urosodeoxycholic acid among the three groups (P>0.05).

CONCLUSIONSFecal secondary colonic bile acids, deoxycholic acid and lithocholic acid, are in decrease in children with HSP at the acute stage, which may be involved in the pathogenesis and treatment outcomes of HSP.

Bile Acids and Salts ; analysis ; Biomarkers ; analysis ; Child ; Feces ; chemistry ; Female ; Humans ; Male ; Purpura, Schoenlein-Henoch ; diagnosis ; therapy

OBJECTIVETo evaluate the detection of fecal PPAR-delta and COX-2 mRNA in screening of colorectal cancer.

METHODSFifty-one patients with colorectal cancer and 21 healthy controls were included in this study. Total RNA was isolated from the fecal samples. Expression of PPAR-delta and COX-2 mRNA was determined by RT-PCR, and its value in screening of colorectal cancer was investigated.

RESULTSThe positive detection rate of fecal PPAR-delta and COX-2 mRNA in colorectal cancer patients was significantly higher than that in healthy controls. In 47 colorectal cancer patients and 19 healthy controls with positive fecal ACTB mRNA expression, the sensitivity of fecal PPAR-delta mRNA, COX-2 mRNA and PPAR-delta mRNA plus COX-2 mRNA detection in diagnosing colorectal cancer was 76.6%(36/47), 80.9%(38/47) and 91.5%(43/47) respectively; the specificity was 63.2%(12/19), 84.2%(16/19) and 89.5%(17/19) respectively.

CONCLUSIONThe combination detection of fecal PPAR-delta and COX-2 mRNA is effective in screening human colorectal cancer and is better than detection of single marker alone.

Aged ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; Cyclooxygenase 2 ; analysis ; Early Detection of Cancer ; Feces ; chemistry ; Female ; Humans ; Male ; Middle Aged ; PPAR delta ; analysis ; RNA, Messenger ; analysis

OBJECTIVETo develop a multiplex real-time PCR for the detection of Salmonella invasion protein A gene (invA), enterotoxigenic Escherichia coli (ETEC) heat-labile I enterotoxin gene (elt), and Shigella or enteroinvasive E. coli (EIEC) invasive plasmid antigen H gene (ipaH).

METHODSUnder the optimized reaction conditions of the multiplex real-time PCR, invA, elt, and ipaH were determined in 10-fold series of dilution of DNA extracted from Salmonella enterica serovar Typhimurium, ETEC 44815 strain and Shigella F301 strain. The three genes were examined in 90 fecal samples from diarrhea patients using the multiplex real-time PCR. When PCR-positive samples were found, the target strains were isolated and identified.

RESULTSThe detectable concentration for this multiplex real-time PCR was 10 CFU/microl for Shigella F301 strain, 10(2) CFU/microl for S. enterica serovar Typhimurium and ETEC 44815 strain, respectively. Out of 90 fecal samples from diarrhea patients, thirteen were found positive for elt gene (14.4%), and five were found positive for ipaH gene (5.6%). Three E. coli strains positive for elt gene and four E. coli strains positive for ipaH gene were isolated successfully from the PCR-positive samples mentioned above. The detection of invA, elt and ipaH genes was completed in 10 h, which included an enrichment period of 6 h.

CONCLUSIONThe multiplex real-time PCR assay can detect invA, elt, ipaH simultaneously in a single reaction, moreover, it can detect for virulence genes in strains of Salmonella, ETEC, and Shigella or EIEC and screen these pathogens in fecal specimens from patients with diarrhea with a high specificity.

DNA, Bacterial ; analysis ; Diarrhea ; microbiology ; Escherichia coli ; genetics ; Feces ; microbiology ; Humans ; Polymerase Chain Reaction ; methods ; Salmonella ; genetics ; Shigella ; genetics