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.Comparative analysis of metagenomic and 16S rDNA sequencing in gut microbiota of healthy elderly.
Si Qi ZHUANG ; Yi Xin MAO ; Fu Chang DENG ; Yue Yun LUO ; Wan Ying SHI ; Xia LI ; Ya Qiang CAO ; Ji Cheng XU ; Song TANG
Chinese Journal of Preventive Medicine 2022;56(11):1618-1624
Objective: To explore the differences in subsequent analysis between metagenomic and 16Sr DNA sequencing in compositionally characterizing gut microbiota of healthy elderly. Methods: By using a panel study design, five monthly repeated measurements were performed among 76 healthy older people in Jinan City, Shandong Province. Their fecal samples were collected, and genomic DNA was extracted and analyzed through metagenomic and 16Sr DNA sequencing to compare the composition and diversity of gut microbiota. The correlation between species abundance and α diversity was analyzed by Pearson correlation analysis, and the correlation between species abundance and β diversity was determined by Procrustes analysis. Results: The age of 76 participants was (65.07±2.75), and the body mass index was (25.03±2.40) kg/m2. There were 38 males and 38 females. A total of 345 fecal samples were obtained from five monthly repeated measurements . Compared with 16S rDNA sequencing, metagenomic sequencing showed more annotated species at each level. The difference in the number of two sequencing species increased with the decrease of the level. Although there were significant differences in species richness between the two sequencing methods. Their species richness was highly correlated at both phylum (r=0.88, P<0.001) and genus (r=0.77, P<0.001) levels. Bacteroidetes and Firmicutes were the common dominant species. Gut microbiota diversity analysis further showed that there was a significantly positive correlation between α diversity (r=0.70, P<0.001) and β diversities (M2=0.84, P<0.05) in the two groups. Conclusion: The annotation efficiency of metagenomic sequencing is much higher than that of 16S rDNA sequencing. The two sequencing methods are consistent in phylum abundance as well as α diversity.
Male
;
Female
;
Humans
;
Aged
;
Gastrointestinal Microbiome/genetics*
;
DNA, Ribosomal/genetics*
;
Feces
;
Sequence Analysis, DNA
;
Metagenomics
;
RNA, Ribosomal, 16S/genetics*
3.Effects of Tanreqing Capsule on the negative conversion time of nucleic acid in patients with COVID-19: A retrospective cohort study.
Xing ZHANG ; Yan XUE ; Xuan CHEN ; Jia-Min WU ; Zi-Jian SU ; Meng SUN ; Lu-Jiong LIU ; Yi-Bao ZHANG ; Yi-le ZHANG ; Gui-Hua XU ; Miao-Yan SHI ; Xiu-Ming SONG ; Yun-Fei LU ; Xiao-Rong CHEN ; Wei ZHANG ; Qi CHEN
Journal of Integrative Medicine 2021;19(1):36-41
OBJECTIVE:
Traditional Chinese medicine plays a significant role in the treatment of the pandemic of coronavirus disease 2019 (COVID-19). Tanreqing Capsule (TRQC) was used in the treatment of COVID-19 patients in the Shanghai Public Health Clinical Center. This study aimed to investigate the clinical efficacy of TRQC in the treatment of COVID-19.
METHODS:
A retrospective cohort study was conducted on 82 patients who had laboratory-confirmed mild and moderate COVID-19; patients were treated with TRQC in one designated hospital. The treatment and control groups consisted of 25 and 57 cases, respectively. The treatment group was given TRQC orally three times a day, three pills each time, in addition to conventional Western medicine treatments which were also administered to the control group. The clinical efficacy indicators, such as the negative conversion time of pharyngeal swab nucleic acid, the negative conversion time of fecal nucleic acid, the duration of negative conversion of pharyngeal-fecal nucleic acid, and the improvement in the level of immune indicators such as T-cell subsets (CD3, CD4 and CD45) were monitored.
RESULTS:
COVID-19 patients in the treatment group, compared to the control group, had a shorter negative conversion time of fecal nucleic acid (4 vs. 9 days, P = 0.047) and a shorter interval of negative conversion of pharyngeal-fecal nucleic acid (0 vs. 2 days, P = 0.042). The level of CD3
CONCLUSION
Significant reductions in the negative conversion time of fecal nucleic acid and the duration of negative conversion of pharyngeal-fecal nucleic acid were identified in the treatment group as compared to the control group, illustrating the potential therapeutic benefits of using TRQC as a complement to conventional medicine in patients with mild and moderate COVID-19. The underlying mechanism may be related to the improved levels of the immune indicator CD3
Adult
;
Antiviral Agents/therapeutic use*
;
COVID-19/pathology*
;
Capsules
;
DNA, Viral/analysis*
;
Drugs, Chinese Herbal/therapeutic use*
;
Feces/virology*
;
Female
;
Humans
;
Length of Stay
;
Lymphocyte Count
;
Male
;
Medicine, Chinese Traditional/methods*
;
Middle Aged
;
Retrospective Studies
;
SARS-CoV-2/genetics*
;
Severity of Illness Index
;
Treatment Outcome
;
Young Adult
4.Head-to-head comparison of the test performance of self-administered qualitative vs. laboratory-based quantitative fecal immunochemical tests in detecting colorectal neoplasm.
Ming LU ; Yu-Han ZHANG ; Bin LU ; Jie CAI ; Cheng-Cheng LIU ; Hong-Da CHEN ; Min DAI
Chinese Medical Journal 2021;134(11):1335-1344
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
5.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
6.Clinical characteristics of 16 patients with fecal severe acute respiratory syndrome coronavirus 2 nucleic acid-positive.
Yan ZHOU ; Zhiquan XIAO ; Dong CHEN ; Jing GUAN ; Zhiguo ZHOU ; Huirong ZHANG ; Huanfa ZHOU
Journal of Central South University(Medical Sciences) 2020;45(5):560-564
OBJECTIVES:
To analyze the clinical characteristics of fecal severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid-positive in patients with coronavirus dasease 2019 (COVID-19) and to provide a scientific basis for the prevention and control of this disease.
METHODS:
The clinical data of 16 patients with fecal SARS-CoV-2 nucleic acid positive, who hospitalized in the North Branch of the First Hospital of Changsha (Changsha Public Health Rescue Center) from January to February 2020, were retrospectively analyzed. Their clinical manifestations, laboratory data and imaging data were summarized.
RESULTS:
Among the 16 patients, there were 9 males (56.25%) and 7 females (43.75%), the ratio of males to females was 1∶1.29. The age of onset was (43.3±14.6) years. There were 15 patients with contact history of Wuhan, 1 patient with contact history of local patient.Twelve patients were common type (75%), and 4 patients were severe type (25%). Clinical symptoms included fever in 14 patients (87.5%), cough in 12 patients (75%), shortness of breath in 5 patients (31.25%), pharyngalgia in 10 patients (62.5%), fatigue in 7 patients (43.75%), and diarrhea in 4 patients (25%). There were 14 patients (87.5%) with normal or decreased white blood cell count, 11 patients (68.75%) with decreased lymphocyte count, 15 patients (93.75%) with increased erythrocyte sedimentation rate, 13 patients (81.25%) with increased hypersensitivity C-reactive protein, 5 patients (31.25%) with increased procalcitonin, and 8 patients (50%) with increased serum ferritin in peripheral blood, and stool routine was basically normal. Compared with the common type, there was significant difference in the white blood cell and lymphocyte counts in the severe type (<0.01); the infection indicators, such as hypersensitivity C-reactive protein and serum ferritin, were significantly increased, with significant difference (all <0.01); but the procalcitonin and erythrocyte sedimentation rate was not significantly different (both >0.05). Chest CT mainly showed patchy shadows and interstitial changes. According to imaging examination, 4 patients (25%) showed unilateral pneumonia and 12 patients (75%) showed bilateral pneumonia.
CONCLUSIONS
The patients have the clinical symptoms of COVID-19, but gastrointestinal symptoms (such as diarrhea) are more common, and the changes of white blood cell count, lymphocyte count, hypersensitivity C-reactive protein, ferritin are more obvious in severe patients.The positivity of fecal nucleic acid suggests the possibility of digestive tract transmission of SARS-CoV-2, and fecal nucleic acid testing can be used as a routine testing method in clinical practice.
Adult
;
Betacoronavirus
;
isolation & purification
;
C-Reactive Protein
;
analysis
;
China
;
Coronavirus Infections
;
diagnosis
;
physiopathology
;
Diarrhea
;
virology
;
Feces
;
virology
;
Female
;
Ferritins
;
analysis
;
Humans
;
Leukocyte Count
;
Male
;
Middle Aged
;
Pandemics
;
Pneumonia, Viral
;
diagnosis
;
physiopathology
;
Retrospective Studies
7.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*
;
Bombyx/chemistry*
;
C-Reactive Protein/analysis*
;
Feces/chemistry*
;
Gastrointestinal Microbiome
;
High-Throughput Nucleotide Sequencing
;
Medicine, Chinese Traditional
;
Rats
;
Vasopressins/blood*
8.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
;
Pregnancy
;
Prenatal Exposure Delayed Effects
;
Sulfamonomethoxine
;
toxicity
;
TOR Serine-Threonine Kinases
;
metabolism
9.Molecular epidemiology of norovirus in children with acute gastroenteritis in Tianjin, China.
Yu-Lian FANG ; Wei WANG ; Ping WANG ; Yu ZHAO ; Tong-Qiang ZHANG ; Xuan LI ; Chao-Meng FAN ; Shu-Xiang LIN
Chinese Journal of Contemporary Pediatrics 2019;21(2):139-143
OBJECTIVE:
To investigate the molecular epidemiological characteristics of norovirus (NoV) among children with acute gastroenteritis in Tianjin in 2017.
METHODS:
A total of 758 stool specimens were collected from the children with acute gastroenteritis possibly caused by viral infection in Tianjin Children's Hospital between January and December, 2017. Quantitative real-time RT-PCR was used for primary screening of NoV, and conventional RT-PCR was used for gene amplification, sequencing and genotype identification of the VP1 region of capsid protein in positive specimens.
RESULTS:
Among the 758 specimens, 241 (31.8%) were found to have GII NoV. Sequencing of the VP1 region of capsid protein in positive specimens showed that among the 241 specimens with GII NoV, 69 (28.6%) had GII.4 subtype, 51 (21.2%) had GII.3 subtype, 24 (10.0%) had GII.2 subtype, and 18 (7.5%) had other subtypes. There was a significant difference in NoV detection rate between different age groups (P=0.018), and the 1- <4 years group had the highest NoV detection rate (37.3%). There was also a significant difference in NoV detection rate across seasons (P<0.001), and there was a highest NoV detection rate in winter (48.1%). Twenty-seven children (3.6%) had co-infections with NoV and rotavirus.
CONCLUSIONS
NoV is one of the major pathogens of the children with acute gastroenteritis from Tianjin in 2017. GII genotype, especially GII.4 subtype, is the prevalent strain. NoV infection is commonly seen in children less than 4 years and reaches the peak in winter. Some children are found to have co-infections with rotavirus.
Caliciviridae Infections
;
Child
;
China
;
epidemiology
;
Feces
;
Gastroenteritis
;
epidemiology
;
Genotype
;
Humans
;
Molecular Epidemiology
;
Norovirus
;
Phylogeny
;
RNA, Viral
;
Sequence Analysis, DNA
10.Systematic review on the characteristics of acute gastroenteritis outbreaks caused by sapovirus.
Y YU ; X H GUO ; H Q YAN ; Z Y GAO ; W H LI ; B W LIU ; Q Y WANG
Chinese Journal of Epidemiology 2019;40(1):93-98
Objective: To understand the epidemiological and etiological characteristics of outbreaks on acute gastroenteritis caused by sapovirus (SaV) worldwide. Methods: Literature about the outbreaks on acute gastroenteritis caused by SaV were retrieved from the databases including WanFang, CNKI, PubMed and Web of Science after evaluation. Time, geography, setting and population distributions of outbreaks, transmission mode, SaV genotype and clinical characteristics of the patients were analyzed. Results: A total of 34 papers about SaV were included, involving 146 outbreaks occurred between October 1976 and April 2016. In these papers, 138 outbreaks were reported on the related months. All these outbreaks occurred in northern hemisphere. SaV outbreaks occurred all year around, but mainly in cold season, the incidence was highest in December (25 outbreaks) and lowest in in August (2 outbreaks). Most outbreaks were reported by Japan, followed by Canada, the United States of America and the Netherlands. There were 141 outbreaks for which the occurring settings were reported, child-care settings were most commonly reported setting (48/141, 34.04%), followed by long-term care facility (41/141, 29.08%) and hospital (16/141, 11.35%). Clinical symptoms of 1 704 cases in 31 outbreaks were reported, with the most common symptom was diarrhea (1 331/1 704, 78.12%), followed by nausea (829/1 198, 69.20%), abdominal pain (840/1 328, 63.25%), vomiting (824/1 704, 48.36%) and fever (529/1 531, 34.53%). Genotypes of SaV were determined for 119 outbreaks. GⅠ(51/119, 42.86%) and GⅣ (45/119, 37.82%) were predominant. The outbreaks of GⅣ SaV increased suddenly in 2007, and the outbreaks of GⅠ SaV mainly occurred in 2008 and during 2011-2013. Conclusions: SaV outbreaks were reported mainly by developed countries, with most outbreaks occurred in cold season, in child-care settings and long term care facility. GⅠ and GⅣ were the most common genotypes of SaV. Prevention and control of SaV outbreak in China seemed relatively weak, and it is necessary to conduct related training and to strengthen the SaV outbreak surveillance in areas where service is in need.
Caliciviridae Infections/virology*
;
Child
;
China/epidemiology*
;
Disease Outbreaks
;
Feces/virology*
;
Gastroenteritis/virology*
;
Genotype
;
Humans
;
Phylogeny
;
RNA, Viral/genetics*
;
Sapovirus/isolation & purification*
;
Sequence Analysis, DNA

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