1.Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification.
Shou-Yu WANG ; Rui-Yang TAO ; Yi-Ping HOU ; Cheng-Tao LI
Journal of Forensic Medicine 2022;38(6):763-773
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*
2.Identification of metabolites of Yiqi Baoyuan Prescription in rat plasma, bile, urine and feces after oral administration.
Yi-Ying ZHANG ; Li-Ping WU ; Yi-Fan FENG ; Hui REN ; Hong-Jie KANG ; Yue ZHU ; Sheng GUO ; Shu-Lan SU ; Da-Wei QIAN ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2022;47(16):4469-4479
This study was designed to determine the metabolites of Yiqi Baoyuan Prescription(YQBYP) in rats. The ultra-high performance liquid chromatography coupled to time-of-flight mass spectrometry(UPLC-TOF-MS) and mass defect filter(MDF) were employed to analyze the metabolites of YQBYP in rat plasma, bile, urine and feces. Chromatographic separation was conducted on Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) under gradient elution with 0.1% formic acid aqueous solution(A)-acetonitrile(B), and the column temperature was 30 ℃. Electrospray ion(ESI) source was used under positive and negative ion modes, with capillary voltage of 3.0 kV and mass scanning range of m/z 100-1 000. In this experiment, 9 prototype components and 36 metabolites were identified in rat plasma, bile, urine and feces samples. The results showed that the main metabolic pathways of YQBYP in rats involved methylation, demethylation, oxidation, and other phase Ⅰ reactions as well as glucuronidation, sulfation, and other phase Ⅱ reactions. This study provided scientific basis for clarifying the therapeutic material basis of YQBYP and product development.
Administration, Oral
;
Animals
;
Bile/chemistry*
;
Chromatography, High Pressure Liquid/methods*
;
Feces/chemistry*
;
Prescriptions
;
Rats
;
Rats, Sprague-Dawley
3.Re-detectable positive SARS-CoV-2 RNA tests in patients who recovered from COVID-19 with intestinal infection.
Wanyin TAO ; Xiaofang WANG ; Guorong ZHANG ; Meng GUO ; Huan MA ; Dan ZHAO ; Yong SUN ; Jun HE ; Lianxin LIU ; Kaiguang ZHANG ; Yucai WANG ; Jianping WENG ; Xiaoling MA ; Tengchuan JIN ; Shu ZHU
Protein & Cell 2021;12(3):230-235
4.Investigating effect of Faeces Bombycis on intestinal microflora in rats with syndrome of damp retention in middle-jiao by high-throughput sequencing.
Lu WU ; Yan LAI ; Ying WANG ; Li-Hua CHEN ; Yong-Mei GUAN ; Rui-Ge CHU ; Hua-Sheng YANG
China Journal of Chinese Materia Medica 2020;45(3):623-630
The aim of this paper was to investigate the effect of Faeces Bombycis(FB) on the intestinal microflora in rats with syndrome of damp retention in middle-jiao, and to explore its mechanism in regulating intestinal microflora from the perspective of microorganisms contained in FB. The contents of antidiuretic hormone(ADH) and C-reactive protein(CRP) in serum and aquaporin 3(AQP3) in jejunum were determined by enzyme-linked immunosorbent assay(ELISA). Illumina Miseq platform was used for high-throughput sequencing of the rat feces and FB. The ELISA results showed that as compared with the normal control group, the contents of ADH and CRP in the model group were significantly increased(P<0.05), and the content of AQP3 was significantly decreased(P<0.05). After drug administration, the ADH, CRP and AQP3 contents were recovered. Sequencing of rat feces showed that the ACE, Chao1 and Shannon indexes of the intestinal microflora were the lowest in the model group. As compared with the normal control group, the levels from phylum to genus were all significantly changed in model group, and Proteobacteria, Acinetobacter, Anaerobacter, Pseudomonas, and Parabacteroides levels were significantly increased(P<0.05), while Marvinbryantia level was significantly decreased(P<0.05). As compared with the model group, Proteobacteria was significantly decreased in the FB low and high dose groups(P<0.05), and Acinetobacter, Anaerobacter, Pseudomonas, Parabacteroides levels were significantly decreased in the low, medium and high dose groups(P<0.05), while Lachnoanaerobaculum, Intestinimonas and Marvinbryantia were increased significantly in the high dose group(P<0.05). Sequencing analysis of FB showed that the relative abundance of Leclercia, Pantoea, Brachybacterium, Shimwellia, Hartmannibacter, Klebsiella, Serratia, Aurantimonas, Paenibacillus and Bacillus was high in the FB, but they were basically not present or little in the rat feces. In conclusion, FB may play a role in the treatment of "syndrome of damp retention in middle-jiao" by balancing the intestinal microflora, and this effect may be related to the metabolites of microorganisms in the FB.
Animals
;
Aquaporin 3/analysis*
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Bombyx/chemistry*
;
C-Reactive Protein/analysis*
;
Feces/chemistry*
;
Gastrointestinal Microbiome
;
High-Throughput Nucleotide Sequencing
;
Medicine, Chinese Traditional
;
Rats
;
Vasopressins/blood*
5.Effect of chemical ingredients B and C in Kansui stir-baked with vinegar on fecal metabolomics in rats.
Si-Jia GUO ; Dong-Jing JIANG ; Li ZHANG
China Journal of Chinese Materia Medica 2020;45(6):1452-1459
To reveal the toxic mechanism of Kansui stir-baked with vinegar(VEK), conducta comparative study on the metabolites of fecal samples of rats before and after being treated with chemical constituents group B and C(VEKB/VEKC) extracted from VEK by metabolomics approach. The fecal samples of each group were analyzed using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UFLC-Q-TOF-MS). Then the data was processed by principal component analysis(PCA) and partial least square discriminant analysis(OPLS-DA) to screen and identify biomarkers relating to the toxicity of VEK. Besides, t-test was adopted for univariate statistical analysis, so as to study the changes of these biomarkers in drug groups before and after being treated with VEKB/VEKC and explore the effect of VEKB/VEKC on the metabolism of rat feces. Furthermore, the toxic mechanism of VEKB/VEKC was explored based on the results of the metabolic pathway analysis. The results displayed that compared with control group, the metabolism of fecal samples of VEKB and VEKC treated groups show obvious changes, and the VEKB treated group show more significant changes. A total of 16 potential biomarkers and 5 metabolic pathways relating to the toxicity of VEK were found and identified. And the toxicity of VEK might be associated with the disorder of such metabolic pathways as tryptophan metabolism, primary bile acid biosynthesis, amino sugar and nucleotide sugar metabolism, purine metabolism, and degradation of valine, leucine and isoleucine. This study provides a scientific basis for the clinical safety application of VEK.
Acetic Acid
;
Animals
;
Biomarkers
;
Chromatography, High Pressure Liquid
;
Euphorbia/chemistry*
;
Feces/chemistry*
;
Mass Spectrometry
;
Metabolome
;
Rats
6.Follow-up testing of viral nucleic acid in discharged patients with moderate type of COVID-19.
Youjiang LI ; Yingying HU ; Xiaodong ZHANG ; Yuanyuan YU ; Bin LI ; Jianguo WU ; Yingping WU ; Xiaoping XIA ; Jian XU
Journal of Zhejiang University. Medical sciences 2020;49(2):270-274
OBJECTIVE:
To investigate the clinical outcome of patients with moderate type of coronavirus disease 2019 (COVID-19) after discharge by retesting viral nucleic acid.
METHODS:
Seven patients with moderate COVID-19 met the discharge criteria enacted by National Health Commission were quarantined in hospital for 7 days, then continuously quarantined at home for 4 weeks after discharged. During the quarantined period, the symptoms and signs were documented, and sputum or nasal swab and feces samples were collected to test SARS-CoV-2 nucleic acid by RT-PCR method.
RESULTS:
There was no symptoms and signs during the quarantine period in all 7 patients. However, respiratory swabs from 3 patients were confirmed positive of SARS-CoV-2 nucleic acid at 5 to 7 days after they met the discharge criteria.
CONCLUSIONS
There is a relatively high incidence of positive viral nucleic acid in patients met the discharge criteria, and it is suggested that patients met the current discharge criteria should be quarantined in hospital for another 7 days and the follow-up viral testing is necessary.
Betacoronavirus
;
isolation & purification
;
Coronavirus Infections
;
diagnosis
;
Feces
;
chemistry
;
virology
;
Follow-Up Studies
;
Humans
;
Pandemics
;
Patient Discharge
;
statistics & numerical data
;
Pneumonia, Viral
;
diagnosis
;
Quarantine
;
statistics & numerical data
;
RNA, Viral
;
analysis
;
Reverse Transcriptase Polymerase Chain Reaction
;
Time Factors
7.Persistently Upregulated Hippocampal mTOR Signals Mediated by Fecal SCFAs Impair Memory in Male Pups with SMM Exposure in Utero.
Yi Tian ZHU ; Xin Ji LIU ; Kai Yong LIU ; Qiang ZHANG ; Lin Sheng YANG ; Rong WEI ; Jing Jing ZHANG ; Fang Biao TAO
Biomedical and Environmental Sciences 2019;32(5):345-356
OBJECTIVE:
To investigate the molecular mechanisms of the adverse effects of exposure to sulfamonomethoxin (SMM) in pregnancy on the neurobehavioral development of male offspring.
METHODS:
Pregnant mice were randomly divided into four groups: control- (normal saline), low- [10 mg/(kg•day)], middle- [50 mg/(kg•day)], and high-dose [200 mg/(kg•day)] groups, which received SMM by gavage daily during gestational days 1-18. We measured the levels of short-chain fatty acids (SCFAs) in feces from dams and male pups. Furthermore, we analyzed the mRNA and protein levels of genes involved in the mammalian target of rapamycin (mTOR) pathway in the hippocampus of male pups by RT-PCR or Western blotting.
RESULTS:
Fecal SCFA concentrations were significantly decreased in dams. Moreover, the production of individual fecal SCFAs was unbalanced, with a tendency for an increased level of total fecal SCFAs in male pups on postnatal day (PND) 22 and 56. Furthermore, the phosphatidylinositol 3-kinase (PI3k)/protein kinase B (AKT)/mTOR or mTOR/ribosomal protein S6 kinase 1 (S6K1)/4EBP1 signaling pathway was continuously upregulated until PND 56 in male offspring. In addition, the expression of Sepiapterin Reductase (SPR), a potential target of mTOR, was inhibited.
CONCLUSION
In utero exposure to SMM, persistent upregulation of the hippocampal mTOR pathway related to dysfunction of the gut (SCFA)-brain axis may contribute to cognitive deficits in male offspring.
Alcohol Oxidoreductases
;
metabolism
;
Animals
;
Anti-Infective Agents
;
toxicity
;
Fatty Acids, Volatile
;
analysis
;
Feces
;
chemistry
;
Female
;
Hippocampus
;
drug effects
;
metabolism
;
Male
;
Memory
;
drug effects
;
Mice, Inbred ICR
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Pregnancy
;
Prenatal Exposure Delayed Effects
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Sulfamonomethoxine
;
toxicity
;
TOR Serine-Threonine Kinases
;
metabolism
8.Quantification of Panax notoginseng saponins metabolites in rat plasma with in vivo gut microbiota-mediated biotransformation by HPLC-MS/MS.
Yin-Ping GUO ; Man-Yun CHEN ; Li SHAO ; Wei ZHANG ; Tai RAO ; Hong-Hao ZHOU ; Wei-Hua HUANG
Chinese Journal of Natural Medicines (English Ed.) 2019;17(3):231-240
Panax notoginseng saponins (PNS) are the major components of Panax notoginseng, with multiple pharmacological activities but poor oral bioavailability. PNS could be metabolized by gut microbiota in vitro, while the exact role of gut microbiota of PNS metabolism in vivo remains poorly understood. In this study, pseudo germ-free rat models were constructed by using broad-spectrum antibiotics to validate the gut microbiota-mediated transformation of PNS in vivo. Moreover, a high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was developed for quantitative analysis of four metabolites of PNS, including ginsenoside F1 (GF1), ginsenoside Rh2 (GRh2), ginsenoside compound K (GCK) and protopanaxatriol (PPT). The results showed that the four metabolites could be detected in the control rat plasma, while they could not be determined in pseudo germ-free rat plasma. The results implied that PNS could not be biotransformed effectively when gut microbiota was disrupted. In conclusion, gut microbiota plays an important role in biotransformation of PNS into metabolites in vivo.
Animals
;
Anti-Bacterial Agents
;
pharmacology
;
Biotransformation
;
Chromatography, High Pressure Liquid
;
Feces
;
microbiology
;
Gastrointestinal Microbiome
;
drug effects
;
physiology
;
Ginsenosides
;
blood
;
Male
;
Panax notoginseng
;
chemistry
;
Rats, Sprague-Dawley
;
Sapogenins
;
blood
;
Saponins
;
administration & dosage
;
metabolism
;
Tandem Mass Spectrometry
9.Identification of active compound combination contributing to anti-inflammatory activity of Xiao-Cheng-Qi Decoction via human intestinal bacterial metabolism.
Xing-Yan LIU ; Li LI ; Xue-Qing LI ; Bo-Yang YU ; Ji-Hua LIU
Chinese Journal of Natural Medicines (English Ed.) 2018;16(7):513-524
Human intestinal bacteria play an important role in the metabolism of herbal medicines, leading to the variations in their pharmacological profile. The present study aimed to investigate the metabolism of Xiao-Cheng-Qi decoction (XCQD) by human intestinal bacteria and to discover active component combination (ACC) contributing to the anti-inflammatory activity of XCQD. The water extract of XCQD was anaerobically incubated with human intestinal bacteria suspensions for 48 h at 37 °C. A liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) method was performed for identification of the metabolites. In addition, the anti-inflammatory effects of XCQD and biotransformed XCQD (XCQD-BT) were evaluated in vitro with cytokines in RAW264.7 cells induced by lipopolysaccharide (LPS). A total of 51 compounds were identified in XCQD and XCQD-BT. Among them, 20 metabolites were proven to be transformed by human intestinal bacteria. Significantly, a combination of 14 compounds was identified as ACC from XCQD-BT, which was as effective as XCQD in cell models of inflammation. In conclusion, this study provided an applicable method, based on intestinal bacterial metabolism, for identifying combinatory compounds responsible for a certain pharmacological activity of herbal medicines.
Animals
;
Anti-Inflammatory Agents
;
isolation & purification
;
pharmacology
;
therapeutic use
;
Bacteria
;
metabolism
;
Biotransformation
;
Cytokines
;
metabolism
;
Drugs, Chinese Herbal
;
chemistry
;
metabolism
;
Feces
;
microbiology
;
Gastrointestinal Microbiome
;
drug effects
;
Humans
;
Inflammation
;
chemically induced
;
drug therapy
;
Lipopolysaccharides
;
pharmacology
;
Macrophages
;
drug effects
;
metabolism
;
Mice
;
Models, Biological
;
Molecular Structure
;
RAW 264.7 Cells
10.Bitter Melon Powder Protects against Obesity-associated Fatty Liver Disease by Improving Colonic Microenvironment in Rats with High-fat Diet-induced Obesity.
Juan BAI ; Ying ZHU ; Ying DONG
Biomedical and Environmental Sciences 2017;30(8):611-615
This study explored how bitter melon powder (BMP) alters the colonic microenvironment during the development of obesity-associated fatty liver in rats. We observed that BMP effectively inhibited the body weight gain and lipid accumulation in the liver, ameliorated glucose intolerance, and increased the colon weight after an 8-week treatment compared to that in the high-fat diet (HFD) group. BMP significantly decreased fecal water toxicity towards HT-29 cells, as revealed by the cell counting kit (CCK)-8 assay results, and the mRNA expression of Toll-like receptor 4 (TLR4) in colon mucosa. Additionally, gut permeability in the BMP group was restored to normal levels. Finally, BMP alleviated the inflammatory state of the rat colon mucosa and liver tissues as well as the systemic inflammation.
Animals
;
Colon
;
drug effects
;
Dietary Fats
;
administration & dosage
;
adverse effects
;
Fatty Liver
;
etiology
;
prevention & control
;
Feces
;
chemistry
;
HT29 Cells
;
Humans
;
Momordica charantia
;
Obesity
;
complications
;
Powders
;
Rats

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