1.Forensic Validation and Application Evaluation of IDentifier DNA Typing Kit (Yan-Huang34).
Lin-Lin GAO ; Wei XIE ; Su-Juan ZHU ; Da LI ; Qin WANG ; Liang HONG ; You-Ying LI
Journal of Forensic Medicine 2023;39(6):579-585
OBJECTIVES:
To investigate the technical performance of IDentifier DNA typing kit (YanHuang34) and evaluate its forensic application value.
METHODS:
Following the Criterion of Forensic Science Human Fluorescence STR Multiplex Amplification Reagent (GB/T 37226-2018), IDentifier DNA typing kit (YanHuang34) was verified in 11 aspects of species specificity, veracity, sensibility, adaptability, inhibitor tolerance, consistency, balance, reaction condition verification, mixed samples, stability and inter batch consistency. The system efficiency of IDentifier DNA typing kit (YanHuang34) was compared with the PowerPlex® Fusion 6C System, VersaPlex® 27PY System and VeriFilerTM Plus PCR Amplification Kit. The IDentifier DNA typing kit (YanHuang34) was used to detect the swabs of biological samples in daily cases and the STR performances were observed.
RESULTS:
IDentifier DNA typing kit (YanHuang34) had good species specificity, veracity, adaptability, inhibitor tolerance and balance. The sensibility was up to 0.062 5 ng. It was able to detect different types of samples, degraded samples and inhibitor mixed samples. Complete DNA typing could be obtained for samples with the mixture ratio less than 4∶1. The system efficiency of IDentifier DNA typing kit (YanHuang34) was very high, with TDP up to 1-1.08×10-37, CPEtrio and CPEduo up to 1-5.47×10-14 and 1-6.43×10-9, respectively. For the touched biological samples in actual cases, the effective detection rate was 21.05%. The system efficiency of kinship, single parent and full sibling identifications was effectively improved.
CONCLUSIONS
The IDentifier DNA typing kit (YanHuang34) is adaptive to the GB/T 37226-2018 requirements. It can be used for individual identification and paternity identification, and is suitable for application in the field of forensic science.
Humans
;
DNA Fingerprinting
;
Polymerase Chain Reaction
;
Microsatellite Repeats
;
Paternity
;
Species Specificity
2.Study on metabolic dynamics,metabolic enzyme phenotype and species difference of hepatic and intestinal microsome of psoralidin.
Zi-Fei QIN ; Bei-Bei ZHANG ; Han XING ; Pei-le WANG ; Jing YANG ; Xiao-Jian ZHANG ; Zhi-Hong YAO ; Xin-Sheng YAO
China Journal of Chinese Materia Medica 2021;46(13):3410-3421
This study aims to investigate metabolic activities of psoralidin in human liver microsomes( HLM) and intestinal microsomes( HIM),and to identify cytochrome P450 enzymes( CYPs) and UDP-glucuronosyl transferases( UGTs) involved in psoralidin metabolism as well as species differences in the in vitro metabolism of psoralen. First,after incubation serial of psoralidin solutions with nicotinamide adenine dinucleotide phosphate( NADPH) or uridine 5'-diphosphate-glucuronic acid( UDPGA)-supplemented HLM or HIM,two oxidic products( M1 and M2) and two conjugated glucuronides( G1 and G2) were produced in HLM-mediated incubation system,while only M1 and G1 were detected in HIM-supplemented system. The CLintfor M1 in HLM and HIM were 104. 3,and57. 6 μL·min~(-1)·mg~(-1),respectively,while those for G1 were 543. 3,and 75. 9 μL·min~(-1)·mg~(-1),respectively. Furthermore,reaction phenotyping was performed to identify the main contributors to psoralidin metabolism after incubation of psoralidin with NADPH-supplemented twelve CYP isozymes( or UDPGA-supplemented twelve UGT enzymes),respectively. The results showed that CYP1 A1( 39. 5 μL·min~(-1)·mg~(-1)),CYP2 C8( 88. 0 μL·min~(-1)·mg~(-1)),CYP2 C19( 166. 7 μL·min~(-1)·mg~(-1)),and CYP2 D6( 9. 1 μL·min~(-1)·mg~(-1)) were identified as the main CYP isoforms for M1,whereas CYP2 C19( 42. 0 μL·min~(-1)·mg~(-1)) participated more in producing M2. In addition,UGT1 A1( 1 184. 4 μL·min~(-1)·mg~(-1)),UGT1 A7( 922. 8 μL·min~(-1)·mg~(-1)),UGT1 A8( 133. 0 μL·min~(-1)·mg~(-1)),UGT1 A9( 348. 6 μL·min~(-1)·mg~(-1)) and UGT2 B7( 118. 7 μL·min~(-1)·mg~(-1)) played important roles in the generation of G1,while UGT1 A9( 111. 3 μL·min~(-1)·mg~(-1)) was regarded as the key UGT isozyme for G2. Moreover,different concentrations of psoralidin were incubated with monkey liver microsomes( MkLM),rat liver microsomes( RLM),mice liver microsomes( MLM),dog liver microsomes( DLM) and mini-pig liver microsomes( MpLM),respectively. The obtained CLintwere used to evaluate the species differences.Phase Ⅰ metabolism and glucuronidation of psoralidinby liver microsomes showed significant species differences. In general,psoralidin underwent efficient hepatic and intestinal metabolisms. CYP1 A1,CYP2 C8,CYP2 C19,CYP2 D6 and UGT1 A1,UGT1 A7,UGT1 A8,UGT1 A9,UGT2 B7 were identified as the main contributors responsible for phase Ⅰ metabolism and glucuronidation,respectively. Rat and mini-pig were considered as the appropriate model animals to investigate phase Ⅰ metabolism and glucuronidation,respectively.
Animals
;
Benzofurans
;
Coumarins
;
Dogs
;
Glucuronides
;
Glucuronosyltransferase/metabolism*
;
Kinetics
;
Mice
;
Microsomes, Liver/metabolism*
;
Phenotype
;
Rats
;
Species Specificity
;
Swine
;
Swine, Miniature/metabolism*
3.Interspecies Phylogenetic Analysis of
Xiao Lei LONG ; Shi Ping WANG ; Shuai Feng ZHOU ; Zheng Xiang LI ; Shi Feng ZHUANG ; Zhuo HE
Biomedical and Environmental Sciences 2021;34(11):881-890
Objective:
This study aims to investigate the infection of
Method:
Infection of the definitive human host and intermediate fish host by
Results:
In 2016-2020, the average population infection rate of Hunan was 1.38%, while in Tongdao County the rate was up to 26.90%, and the highest fish infection rate was detected in Qiyang County (99.44% in the dorsal fin of
Conclusion
The systematically study of
Animals
;
Cat Diseases/parasitology*
;
Cats
;
China/epidemiology*
;
Clonorchiasis/veterinary*
;
Clonorchis sinensis/genetics*
;
Dog Diseases/parasitology*
;
Dogs
;
Fish Diseases/parasitology*
;
Fishes
;
Humans
;
Incidence
;
Prevalence
;
Species Specificity
4.Molecular Detection and Identification of
Hong Qing ZHAO ; Pei Pei LIU ; Feng XUE ; Miao LU ; Xin Cheng QIN ; Kun LI
Biomedical and Environmental Sciences 2021;34(12):1020-1023
5.Development and Forensic Application of EX20+30Y Kit.
Xu-Ming ZHOU ; Deng-Xing GAO ; Fa-Yuan LI
Journal of Forensic Medicine 2021;37(5):639-645
OBJECTIVES:
To develop a multiplex PCR amplification system (EX20+30Y for short) of 19 autosomes, 30 Y-STR loci plus the gender indicator, and evaluate its forensic application value.
METHODS:
With the six-color fluorescence labeling technology, a multiplex amplification system of 19 autosomal STR loci and 30 Y-STR loci plus the gender indicator was constructed. Blood samples from 210 unrelated individuals, 69 daily case samples and standard samples 9948 and 9947A were collected for loci detection and analysis. The EX20+30Y multiplex amplification system was evaluated by its sensitivity, mixed sample detection ability, species specificity, balance, direct amplification ability, sample applicability and anti-inhibition ability.
RESULTS:
Multiplex amplification of blood samples from 210 unrelated individuals by the detection system obtained accurate genotyping results. The detection sensitivity of standard samples was 0.125 ng and the species specificity was high. The 69 samples from daily cases were genotyped correctly. Moreover, standard sample 9948 could be accurately genotyped even if the samples contained a certain concentration of inhibitors.
CONCLUSIONS
The multiplex amplification system established in this study can conduct combined examination of 19 autosomes, 30 Y-STR loci plus the gender indicator with accurate genotyping and high sensitivity. It has a good forensic application prospect.
Chromosomes, Human, Y/genetics*
;
DNA Fingerprinting/methods*
;
Forensic Medicine/methods*
;
Humans
;
Microsatellite Repeats
;
Multiplex Polymerase Chain Reaction
;
Species Specificity
6.Construction of a highly efficient synthetic lycopene engineered Saccharomyces cerevisiae.
Ling SUN ; Junhua WANG ; Wei JIANG ; Youran LI ; Liang ZHANG ; Zhongyang DING ; Zhenghua GU ; Guiyang SHI ; Sha XU
Chinese Journal of Biotechnology 2020;36(7):1334-1345
Lycopene, as a high value-added terpene compound, has been widely concerned by researchers at home and abroad. Firstly, the ability of lycopene synthesis of Saccharomyces cerevisiae model strains S288c and YPH499 was analyzed and compared. The results showed that YPH499 was more suitable for lycopene synthesis as yeast chassis. Subsequently, the effects of constitutive promoters GPDpr, TEF1pr and inducible promoters GAL1pr, GAL10pr on Lycopene synthesis were compared. The results showed that when GPDpr and TEF1pr were used as promoters of crtE, crtB and crtI in lycopene synthesis pathway, the production of lycopene was 15.31 mg/L after 60 h fermentation in shaking flask. When GAL1pr and GAL10pr were used as promoters, the production was 123.89 mg/L, which was 8.09 times higher. In addition, the methylvaleric acid (MVA) pathway was further modified to overexpress the key enzyme gene of N-terminal truncation, tHMG1 (3-hydroxy-3-methylglutaryl coenzyme A reductase). The lycopene production was 265.68 mg/L, and the yield per cell was 72.79 mg/g. The Saccharomyces cerevisiae strain designed and constructed in this study can express lycopene in high yield per cell, thus could be used in the industrial production of lycopene after further construction and optimization.
Biosynthetic Pathways
;
genetics
;
Fermentation
;
Industrial Microbiology
;
Lycopene
;
metabolism
;
Saccharomyces cerevisiae
;
genetics
;
metabolism
;
Species Specificity
7.Genetic Diversity, Antimicrobial Resistance, and Virulence Genes of Isolates from Clinical Patients, Tap Water Systems, and Food.
Shuang MENG ; Yong Lu WANG ; ChenGeng LIU ; Jing YANG ; Min YUAN ; Xiang Ning BAI ; Dong JIN ; Jun Rong LIANG ; Zhi Gang CUI ; Juan LI
Biomedical and Environmental Sciences 2020;33(6):385-395
Objective:
This study aimed to evaluate the genetic diversity, virulence, and antimicrobial resistance of isolates from clinical patients, tap water systems, and food.
Methods:
Ninety isolates were obtained from Ma'anshan, Anhui province, China, and subjected to multi-locus sequence typing (MLST) with six housekeeping genes. Their taxonomy was investigated using concatenated sequences, while their resistance to 12 antibiotics was evaluated. Ten putative virulence factors and several resistance genes were identified by PCR and sequencing.
Results:
The 90 isolates were divided into 84 sequence types, 80 of which were novel, indicating high genetic diversity. The isolates were classified into eight different species. PCR assays identified virulence genes in the isolates, with the enterotoxin and hemolysin genes , , , and found in 47 (52.2%), 13 (14.4%), 22 (24.4%), and 12 (13.3%) of the isolates, respectively. The majority of the isolates (≥ 90%) were susceptible to aztreonam, imipenem, cefepime, chloramphenicol, gentamicin, tetracycline, and ciprofloxacin. However, several resistance genes were detected in the isolates, as well as a new variant.
Conclusions
Sequence type, virulence properties, and antibiotic resistance vary in isolates from clinical patients, tap water systems, and food.
Aeromonas
;
drug effects
;
genetics
;
isolation & purification
;
pathogenicity
;
Anti-Bacterial Agents
;
pharmacology
;
China
;
Drinking Water
;
microbiology
;
Drug Resistance, Bacterial
;
Food Microbiology
;
Genetic Variation
;
Gram-Negative Bacterial Infections
;
microbiology
;
Species Specificity
;
Virulence
9.Genome Size Evolution Mediated by Gypsy Retrotransposons in Brassicaceae.
Shi-Jian ZHANG ; Lei LIU ; Ruolin YANG ; Xiangfeng WANG
Genomics, Proteomics & Bioinformatics 2020;18(3):321-332
The dynamic activity of transposable elements (TEs) contributes to the vast diversity of genome size and architecture among plants. Here, we examined the genomic distribution and transposition activity of long terminal repeat retrotransposons (LTR-RTs) in Arabidopsis thaliana (Ath) and three of its relatives, Arabidopsis lyrata (Aly), Eutrema salsugineum (Esa), and Schrenkiella parvula (Spa), in Brassicaceae. Our analyses revealed the distinct evolutionary dynamics of Gypsyretrotransposons, which reflects the different patterns of genome size changes of the four species over the past million years. The rate of Gypsy transposition in Aly is approximately five times more rapid than that of Ath and Esa, suggesting an expanding Aly genome. Gypsy insertions in Esa are strictly confined to pericentromeric heterochromatin and associated with dramatic centromere expansion. In contrast, Gypsy insertions in Spa have been largely suppressed over the last million years, likely as a result of a combination of an inherent molecular mechanism of preferential DNA removal and purifying selection at Gypsy elements. Additionally, species-specific clades of Gypsy elements shaped the distinct genome architectures of Aly and Esa.
Brassicaceae/genetics*
;
Evolution, Molecular
;
Genome Size
;
Genome, Plant
;
Genomics
;
Phylogeny
;
Retroelements
;
Species Specificity
10.Isolation of Penicillium expansum WH-3 for the production of L(+)-tartaric acid.
Wen-Na BAO ; Yi CHEN ; Hong-Xiu LIAO ; Hang CHEN ; Shi-Wang LIU ; Yong LIU
Journal of Zhejiang University. Science. B 2020;21(10):835-840
The L(+)-form of tartaric acid (L(+)-TA) exists extensively in nature, and is widely used in the food, chemical, textile, building, and pharmaceutical industries (Su et al., 2001). The main method for L(+)-TA production is microbial transformation by cis-epoxysuccinate hydrolase (CESH), which can catalyze the asymmetric hydrolysis of cis-epoxysuccinic acid or its salts to TA or tartrate (Bao et al., 2019). Seventeen species containing CESH have been isolated so far. However, most species for L(+)-TA production have been reported from bacteria (Xuan and Feng, 2019). The only fungus isolated from soil by our lab recently, that could be used as catalyst for the process under acidic condition, is Aspergillus niger WH-2 (Bao et al., 2020). In order to find strains with new characteristics, this study attempted to isolate a new CESH source from fungi and investigate its application value.
Aspergillus niger/metabolism*
;
Biomass
;
Catalysis
;
Fermentation
;
Hydrogen-Ion Concentration
;
Hydrolases/chemistry*
;
Hydrolysis
;
Industrial Microbiology
;
Magnetic Resonance Spectroscopy
;
Penicillium/metabolism*
;
Phylogeny
;
Soil
;
Species Specificity
;
Stereoisomerism
;
Tartrates/chemistry*
;
Temperature
;
Textiles

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