1.Progress in the biomarker discovery for drug-induced liver injury.
Lei-yan HE ; Yao-xue GUO ; Chun LI ; Ye DENG ; Qi-zhi ZHANG ; Wen-xing PENG
Acta Pharmaceutica Sinica 2015;50(8):959-965
The leading cause of drug withdrawal from market and clinical trials failure is drug-induced liver injury (DILI). Varying clinical, histological and laboratory features of DILI, as well as undefined underlying mechanisms, hinder patients to be diagnosed in the early-stage of the disease and receive effective treatments. Conventional indicators, like serum transaminases and bilirubin, have inevitable limitations referring to sensitive prediction and specific detection of DILI. In order to reduce the occurrence of DILI, researchers have attempted to discover potential biomarkers with higher specificity and sensitivity from blood and urine in recent years. This article aims to review recent advances in biomarkers of DILI.
Biomarkers
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blood
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urine
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Chemical and Drug Induced Liver Injury
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diagnosis
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Humans
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Sensitivity and Specificity
2.Construction,expression and identification of eukaryotic expression vector carrying Schistosoma japonicum gene coding lysophospholipase
Gong-si, FANG ; Yong, YAO ; Li-wen, WANG ; Xue-long, WANG
Chinese Journal of Endemiology 2011;30(4):364-367
Objective Schistasoma japonicum(S.japonicum)lysophospholipase gene(Sjl539)from cDNA of S japonicum adult worms was amplified and subcloned into eukaryotic expression vector pcDNA3.1(+)for expression of recombinant antigen and immunogenicity analysis.Methods Total RNA of S.japonicum was extracted to generato cDNA by RT-PCR.The Sj1539 gent was amplified.The DNA fragment was subcloned into eukaryofic expression vector pcDNA3.1(+)following insertion and amplification in pGEM-T.The recombinant plasmid was transfected into human cervical carcinoma cell strain(Hela cells)and expression products were identified by Western blotting.Results The size of PCR product was approximately 684 bp.It was confirmed that Sj1539 gene had been inserted successfully by the recombinant plasmid digested with two enzymes and PCR.It was verified that the expression product could react with S.japonicum-infected rabbit serum by Western blotting and the molecular weight was approximately 25×103.Conclusions The eukaryotie expression vector carrying Sj1539 gene has been established and the expression product has been obtained.
3.Relation of social adaptability to theory of mind and executive function in children with autism
Li YAO ; Jing LIU ; Xue LI ; Wen YANG ; Yujing ZHOU ; Hui WANG
Chinese Mental Health Journal 2015;(9):692-696
Objective:To explore the relation of social adaptability to theory of mind (Tom)and executive function (EF)in Chinese children with autism.Method:Thirty-eight children aged 6.0 -12.9 years and meeting the Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-IV)criteria for autism were re-cruited.They were assessed with the Children's Adaptive Behavior Rating Scale,Chinese Wechsler Intelligence Scale for Children (C-WISC),Tom test and EF task,then their adaptive quotient (ADQ),intelligence quotient, scores of theory of mind and executive function were obtained.Results:In this sample,the ADQ was (85.4 ± 20.9),the T scores of independent factor,cognitive factor and social self-control factor were separately (40.2 ± 17.3),(43.3 ±15.7)and (42.0 ±16.0).The median total intelligence quotient was (102.1 ±24.0).After con-trolling for age and intelligence quotient,the score of first false belief understanding was positively correlated with social responsibility subsclale score and social self-control factor score (r =0.50,0.45,Ps <0.05).In executive function test,scores of immediate and delayed recall for details of Rey complex figure recall task were negatively correlated with social responsibility subscale score (r =-0.46,-0.47,Ps <0.05).In Stroop tests,the second test time,the forth test time,the forth test error number and alpha numeric connection error number were negatively cor-related with economic activity subscale score (r =-0.63,-0.57,-0.45,-0.57,Ps <0.05).The second test time of Stroop was negatively correlated with the scores of individual choose subscale and social self-control factor (r =-0.46,-0.50,Ps <0.05).Conclusion:It suggests that social adaptability may be related to theory of mind and executive function.The autistic children with better theory of mind and executive function have better social a-daptability.
4.A novel 6-dye STR 25-plex DNA typing system for forensic casework and databases
Xue BAI ; Yiren YAO ; Jian YE ; Wen SONG ; Guangshu DING
Chinese Journal of Forensic Medicine 2020;35(3):258-264
To develop and validate a novel 6-dye STR(short tandem repeat) 25-plex DNA typing system for forensic DNA profiling and databases. In this study, a novel STR 25-plex DNA typing system that includes 24 autosomal STRs (D1S1656, D2S1338, D2S441, D3S1358, D5S818, D6S1043, D7S820, D8S1179, D10S1248, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11, D22S1045, CSF1PO, FGA, Penta D, Penta E, TH01, TPOX, vWA, D11S4463) and Amelogenin was developed. Validation studies demonstrated the sensitivity, accuracy, and reproducibility of our novel STR 25-plex DNA typing system. The sensitivity of the STR 25-plex DNA typing system was demonstrated by the ability to obtain complete profiles from as little as 0.125ng of human DNA. Specificity testing was demonstrated by the lack of cross-reactivity to a variety of commonly encountered animal species and microbial pool. For stability testing, full profiles were obtained with humic acid concentration ≤60ng/μL and hematin ≤600μM. For forensic evaluation, the selected 24 autosomal STRs followed the Hardy–Weinberg equilibrium. Since 24 autosomal STRs were independent from one another, PM (Probability matching) was 3.5434×10-28, TDP (Total Probability of Discrimination Power) was 0.999999999999999999999999969863, and CEP (Cumulative probability of exclusion) was 0.99999999375. The new STR 25-plex typing system is sensitive, reproducible, and stable, therefore it is highly applicable for use in national DNA database and can help to facilitate international data sharing.
5.Construction of a 15-plex Rapid STR Multiplex Amplification System.
Jun-Ping HAN ; Jing SUN ; Yuan OU ; Peng LIU ; Jian YE ; Wen-wen ZHAO ; Xue-qian WANG ; Yi-wen ZHANG ; Yao LIU ; Cai-xia LI
Journal of Forensic Medicine 2016;32(1):49-53
OBJECTIVE:
To establish a 15-plex rapid STR multiplex amplification system.
METHODS:
Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives.
RESULTS:
Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure.
CONCLUSION
The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Alleles
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Chromosome Mapping
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DNA/genetics*
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DNA Fingerprinting/methods*
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Forensic Genetics/methods*
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Humans
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Microsatellite Repeats/genetics*
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Polymerase Chain Reaction/methods*
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Racial Groups/genetics*
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Sensitivity and Specificity
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Tandem Repeat Sequences
6.Effect of position on oxygenation in neonates after weaning from mechanical ventilation.
Wen-Xiu YAO ; Xin-Dong XUE ; Jian-Hua FU
Chinese Journal of Contemporary Pediatrics 2008;10(2):121-124
OBJECTIVEWhat is the best suitable position for neonates who were weaned from mechanical ventilation has not been identified. This study aimed to evaluate the effect of the supine and prone positions on oxygenation in neonates within 6 hrs after weaning from mechanical ventilation.
METHODSSixty neonates who were weaned from mechanical ventilation were randomly given prone or supine position (n=30 each). They all received oxygen inspiration and SPO2 was maintained in a normal range by adjusting the oxygen flow rate (FiO2). Blood PaO2 and PaCO2 levels were measured 1 and 6 hrs after weaning and then the alveolodouble ended arrowarterial oxygen partial pressure difference (A-aDO2), respiratory index and oxygenation index were calculated.
RESULTSMean FiO2 used in the prone position group was significantly lower than that in the supine position group 1 and 6 hrs after weaning (P<0.01). The value of A-aDO2 in the prone position group 1 hr (171.06+/-86.55 vs 253.62+/-71.56; P<0.01) and 6 hrs after weaning (105.85+/-78.18 vs 208.48+/-86.80; P<0.01) were significantly lower than that in the supine position group. The respiratory index in the prone position group 1 and 6 hrs after weaning (2.16+/-1.24 and 1.35+/-1.11) was also reduced compared to 3.74+/-1.68 and 3.65+/-1.28 in the supine position group (P<0.01). In contrast, PaO2 in the prone position group 1 hr (88.70+/-32.65 vs 73.43+/-17.68; P<0.01) and 6 hrs (84.10+/-13.95 vs 70.20+/-20.27; P<0.01) after weaning was significantly higher than that in the supine position group. The oxygenation index in the prone position group 1 and 6 hrs after weaning (213.49+/-88.96 and 275.23+/-108.83) increased significantly compared to 141.54+/-43.25 and 160.62+/-63.03 in the supine position (P<0.01).
CONCLUSIONSThe prone position is better than the supine position for the improvement of oxygenation within 6 hrs after weaning from mechanical ventilation in neonates.
Female ; Humans ; Infant, Newborn ; Male ; Oxygen ; metabolism ; Posture ; Respiration, Artificial
7.Based on supramolecular chemistry to explore the scientific connotation of the compatibility between licorice and the insoluble mineral medicine gypsum
Yao-zhi ZHANG ; Wen-min PI ; Lin-ying WU ; Lu-ping YANG ; Shu-chang YAO ; Xiang ZHANG ; Xue-mei HUANG ; Peng-long WANG
Acta Pharmaceutica Sinica 2024;59(4):1048-1056
Licorice-gypsum (gancao-shigao, GC-SG) drug pair was used as the research object, using supramolecular chemistry to explore the scientific connotation of combining herbal medicine GC with insoluble mineral medicine SG in clinical application of traditional Chinese medicine. ① The Tyndall effect, microscopic morphology and particle size of the single and co-decocted of GC and SG were observed, the paste content and conductivity were determined, and the interaction between GC and SG was detected by isothermal titration calorimetry (ITC) and infrared absorption spectroscopy (IR). ② Calcium chloride (CaCl2), a soluble calcium salt of equal gypsum quality, was used instead of SG with GC for co-decocting to explore the effect of calcium salt content on the water decocting, and the characteristics were combined with the Tyndall effect, microscopic morphology, paste content and conductivity. ITC and IR techniques were used to detect the interaction between the two, and the interaction between them was detected by ITC and IR. The zeta potential and ultraviolet-visible spectrophotometry (UV-vis) of GC-SG and GC-CaCl2 co-decoction were compared, and the inorganic and organic components in the co-decoction were detected by inductively coupled plasma optical emission spectrometer (ICP-OES) and high performance liquid chromatography (HPLC). The results showed: ① Compared with the liquid phase of single decoction, GC-SG co-decoction had more obvious Tyndall effect, and showed uniform spherical nanoparticles under electron microscope. Physical characterization results such as paste content and conductivity showed that co-decoction promoted the dissolution of each other's components; ITC and IR results showed that there was strong interaction between GC and SG, which preliminatively indicated that GC and SG co-decoction promoted the formation of uniform and stable supramolecular system of traditional Chinese medicine. ② When soluble calcium salt was used to substitute insoluble SG with GC for co-decocting, a stronger but astigmatic light path appeared than single decocting solution, the zeta potential was reduced, and a large number of accumulated polymers were formed. The results of paste content and conductivity showed that the dissolution of the co-decocting component was reduced than the single decocting component. ITC, UV-vis and IR results showed that there was interaction between GC with Ca2+ and SG. The formation of polysink indicated that a large amount of soluble calcium salt would destroy the stability of supramolecular Chinese medicine. The results of ICP-OES and HPLC showed that the glycyrrhizic acid (GA) content of the former lower than the latter, which was related to the formation of a large number of polycondensates with the increase of Ca2+ concentration and the decrease of the dissolution of GA and other active ingredients. This study indicates that the compatibility of GC and SG can form a uniform and stable supramolecular system of traditional Chinese medicine. Calcium salt, the main component of SG, is taken as the starting point. Excessive soluble Ca2+ can promote the aggregation of active ingredients such as GA, so as to reveal the scientific connotation of the compatibility of GC and SG, an insoluble mineral medicine.
8.Effects of interleukin-1β on mineralization potential of dental pulp stem cells
Xue-Chao YANG ; Si-Yuan ZHANG ; Ming-Wen FAN ; Xin LI ; Tian LIU ; Yao YAO
Chinese Journal of Stomatology 2011;46(7):406-411
Objective To investigate the effects of pro-inflammatory cytokine interleukin-1β(IL-1β)on mineralization potential of dental pulp stem cells(DPSC). Methods Rat DPSC were cultured in vitro and randomly divided into three groups, IL-1β(10 μg/L),osteogenic inductive medium and non-osteogenic inductive medium. After 3, 7, and 12 days of treatment, the cultures were evaluated for cell proliferation and calcium deposit. Real-time polymerase chain reaction was used to detect the gene expression levels of osteocalcin(OC), bone sialoprotein(BSP), dentin sialophosphoprotein(DSPP) and dentin matrix protein 1(DMP-1). In vivo test, after 3 day′s treatment with IL-1β, the cell-scaffold complexes were implanted subcutaneously in mice for 8 weeks. Histological analysis was performed to evaluate hard tissue formation. Results In vitro test, after 3-day′s treatment, IL-1β improved cell proliferation to 137.22 DNA μg/L and cell viability becomes(97.12±7.18)% of control. The gene expression levels of OC, BSP, DSPP and DMP-1 are(378.19±16.22)%,(427.12±18.22)%,(247.19±10.11)% and(198.29±10.23)% respectively. The results of IL-1β′s group was notable increased compared with non-osteogenic induction medium and the statistical differences are significant. IL-1β induced the odontogenic differentiation of DPSC. However, these effects tended to continuously decrease with treatment time. Histological analysis demonstrated that in the group treated with IL-1β hard tissue was markedly formed in vivo. Conclusions IL-1β may induce the mineralization of DPSC and play an important role in host defenses and tissue repair.
9.Analysis of interspecies adherence of oral bacteria using a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis profiling.
Ren-ke WANG ; Xue-song HE ; Wei HU ; Renate LUX ; Ji-yao LI ; Xue-dong ZHOU ; Wen-yuan SHI
International Journal of Oral Science 2011;3(2):90-97
Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol x L(-1) glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.
Adult
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Animals
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Bacterial Adhesion
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DNA, Bacterial
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analysis
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Denaturing Gradient Gel Electrophoresis
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Fusobacterium nucleatum
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physiology
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Humans
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Membranes, Artificial
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Mice
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Microbial Interactions
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physiology
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Polymerase Chain Reaction
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Protein Binding
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Saliva
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microbiology
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Streptococcus mutans
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physiology
10.Effects of vitamin D analogue EB1089 on proliferation and apoptosis of hepatic carcinoma cells.
Wen-jing LUO ; Jing-yuan CHEN ; Wen XU ; Fang ZHAO ; Yao-ming CHEN ; Xue-feng SHEN
Chinese Journal of Preventive Medicine 2004;38(6):415-418
OBJECTIVEThis study aimed at investigating the effects of vitamin D analogue EB1089 on the proliferation and apoptosis of hepatic carcinoma cells.
METHODSHepatic carcinoma cell strain G(2) (Hep-G(2)) in which prominent vitamin D receptor (VDR) mRNA could be expressed and the cell strain T (HCC-T) negative in VDR gene expression were incubated in culture media with 100 nmol/L, 10 nmol/L and 1 nmol/L EB1089 for 2 d, 4 d and 6 d, respectively. Survival and proliferation of the cells were detected by blue tetrazolium colorimetric test and plate clone-forming test, the VDR mRNA expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and apoptosis of the cells was detected by flow cytometry (FCM) and electron microscopy.
RESULTSEB1089 could inhibit the proliferation of hepatocellular cell line Hep-G(2) that expressed prominent vitamin D receptor mRNA, the inhibitory rate is 17.5% approximately 72.1%. On the other hand, EB1089 had no anti-proliferative effect on hepatocellular cell line HCC-T in which the gene expression of vitamin D receptors was negative. The electron microscope results showed that EB1089 could induce apoptosis of hepatocarcinoma cells and the percentages of apoptotic cells measured by flow cytometer was 21.4%. Cell cycle progression was blocked at G(1) phase with EB1089.
CONCLUSIONEB1089 could inhibit proliferation of human Hep-G(2), probably through VDR, and induce apoptosis of the cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Calcitriol ; analogs & derivatives ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Division ; drug effects ; Humans ; Liver Neoplasms ; pathology ; Tumor Cells, Cultured