OBJECTIVES: To investigate the regulatory role of the NLRP3 signaling pathway in hepatocyte pyroptosis in nonalcoholic steatohepatitis (NASH) under hypoxia. METHODS: Twenty-four male C57BL/6 mice were randomized equally into hypoxic control (A), hypoxic NASH model (B), hypoxic NASH+NLRP3 inhibitor (C), and hypoxic NASH+caspase-1 inhibitor (D) groups. In groups B-D, the mice were fed a methionine choline-deficient (MCD) diet under hypoxic conditions (to simulate a 5000 m altitude) for 6 weeks; the mice in groups C and D received intraperitoneal injections of the respective inhibitors every other day. RESULTS: Compared with those in group A, the mice in group B showed significantly elevated serum levels of FBG, TC, TG, ALT and AST, increased liver lipid content, inflammatory cell infiltration and collagen fiber deposition, and enhanced hepatic expressions of NLRP3, caspase-1, IL-1β and GSDMD proteins, with obvious swelling, cristae breakage, vacuolization, and outer membrane disruption of the mitochondria, ribosome loss in the cytoplasm, destruction of the nuclear membrane, and pathological changes of the rough endoplasmic reticulum. Treatment with NLRP3 inhibitor and caspase-1 inhibitor both significantly lowered serum levels of TC, TG, ALT and AST (but without significantly affecting FBG) in the mouse models, and reduced liver lipid content, inflammatory cell infiltration, collagen deposition, and expression levels of NLRP3, caspase-1, GSDMD and IL-1β. The treatments also significantly improved pathological changes in the mitochondria, ribosomes and endoplasmic reticulum in liver tissues of the mice. CONCLUSIONS NLRP3 signaling pathway plays a key role in promoting hepatocyte pyroptosis in NASH mice under hypoxic condition, and inhibiting this pathway can effectively reduce liver inflammation, suggesting its potential as a therapeutic target for NASH treatment.
Animals ; Non-alcoholic Fatty Liver Disease/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Pyroptosis ; Mice, Inbred C57BL ; Male ; Hepatocytes/pathology* ; Signal Transduction ; Mice ; Hypoxia/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

Objective To investigate the effect of a nucleic acid purifier combined with a vacuum concentrator on DNA recovery rate.Methods Using 39 tubes of 007 standard samples,two 30 μL samples were taken from each tube.Samples were purified using the PrepFiler ExpressTM BTA Purification Kit on the AutoMate ExpressTM purifier,with elution volumes of 30 μL and 200 μL.Samples with 200 μL eluent were concentrated to 30 μL using a trace DNA sample vacuum concentrator.DNA solution concentrations were measured,and recovery rates were calculated and compared.Results The measured DNA concentration of the 007 standard samples was 0.08～0.15(0.11±0.02)ng/μL,showing a significant difference from the kit-labeled concentration of 0.1 ng/μL(t=3.88,P<0.01).The DNA solution concentration in the purified and concentrated group(200 μL eluent concentrated to 30 μL)[0.05～0.13(0.08±0.02)ng/μL]was significantly higher than the purified group(30 μL eluent)[0.02～0.08(0.05±0.01)ng/μL](t=21.20,P<0.01).The DNA recovery rate of the purified and concentrated group[54.55%～92.86%(75.41±10.12)%]was substantially higher than the purified group[25.00%～54.55%(39.79±7.13)%](t=19.79,P<0.01).Conclusion Utilizing a large-volume elution system in nucleic acid purification can significantly enhance DNA recovery rate.Combining this method with a vacuum concentrator effectively increases DNA solution concentration,thereby improving DNA testing success rates.

Objective To investigate the effect of a nucleic acid purifier combined with a vacuum concentrator on DNA recovery rate.Methods Using 39 tubes of 007 standard samples,two 30 μL samples were taken from each tube.Samples were purified using the PrepFiler ExpressTM BTA Purification Kit on the AutoMate ExpressTM purifier,with elution volumes of 30 μL and 200 μL.Samples with 200 μL eluent were concentrated to 30 μL using a trace DNA sample vacuum concentrator.DNA solution concentrations were measured,and recovery rates were calculated and compared.Results The measured DNA concentration of the 007 standard samples was 0.08～0.15(0.11±0.02)ng/μL,showing a significant difference from the kit-labeled concentration of 0.1 ng/μL(t=3.88,P<0.01).The DNA solution concentration in the purified and concentrated group(200 μL eluent concentrated to 30 μL)[0.05～0.13(0.08±0.02)ng/μL]was significantly higher than the purified group(30 μL eluent)[0.02～0.08(0.05±0.01)ng/μL](t=21.20,P<0.01).The DNA recovery rate of the purified and concentrated group[54.55%～92.86%(75.41±10.12)%]was substantially higher than the purified group[25.00%～54.55%(39.79±7.13)%](t=19.79,P<0.01).Conclusion Utilizing a large-volume elution system in nucleic acid purification can significantly enhance DNA recovery rate.Combining this method with a vacuum concentrator effectively increases DNA solution concentration,thereby improving DNA testing success rates.

Objective To compare the PCR effect of rapid PCR instrument and common PCR instrument. Methods The concentration of 9947A standard was diluted to 0.1, 0.05, 0.025, 0.0125, 0.0063 ng/μL, 100 samples from each concentration group to establish PCR reaction system with Identifiler? Plus PCR Amplification Kit, 50 samples of them test with rapid PCR instrument (Speed cycler2 thermocycler), the other 50 samples test with common PCR instrument (9700 thermocycler). Detection of PCR product with 3500xL Genetic Analyser, the STR typing of both groups of each concentration group should be compared. Results The success rate of both thermocyclers have no significant difference (P>0.05); The success number of STR typing of common PCR instrument (13.7±1.0; 11.3±1.5) were higher than rapid PCR instrument (13.1±1.3; 9.9±1.9) when the concentration of 9947A were 0.0125, 0.0063ng/μL (P=0.029; P<0.001); The peak height of DNA typing map obtained from common PCR instrument (18931±4625;13437±3165; 5752±1344) were higher than rapid PCR instrument (16929±4034; 11815±4120; 4865±1401) when the concentration of 9947A were 0.1, 0.05, 0.025ng/μL (P=0.023; P=0.030; P=0.002). Conclusions The rapid PCR instrument could achieve the equal success rate to the common PCR instrument with less time, which revealed that the rapid PCR instrument was suitable for application in practical cases; The quality of STR typing from common PCR instrument may be more higher.

Objective To explore the distributional differences of the gene frequencies of 22 short tandem repeats loci on Y Chromosome(Y?STRs) between offenders with Initiative?aggressive behavior and impulsive?aggressive behavior,and to probe into the genetic factors of initiative?aggressive behavior and im?pulsive?aggressive behavior. Methods Biological samples of 271 offenders with initiative?aggressive behav?ior and 271 offenders with impulsive?aggressive behavior were collected and PCR compound amplification was carried out with the aid of PowerPlex Y23 System. Then the PCR products were subjected to electrophoresis and gene detection with AB3500xL gene analysis system so as to calculate and compare the alleles and haplo?types of 22 Y?STRs gene frequency in the two groups. Results The distribution of allele frequency were sig?nificantly difference in locus DYS437(P=0.022) between two groups,not in the other 21 Y?STRs loci( all P>0.05) . Univarite analysis showed significant differences at allelle 14 in locus DYS437 between both groups ( initiative?aggressive behavior group:69. 37%;impulsive?aggressive behavior group:58. 67%; P=0. 009 ) . Conclusion Loci DYS437 may be associated with aggressive behavior. In the group of aggressive behavior, allelle 14 on locus DYS437 may be the susceptible factor of initiative?aggressive behavior and the resistant factor of impulsive?aggressive.

OBJECTIVETo assess the association of short tandem repeats (STRs) loci with aggressive behaviors of schizophrenia.

METHODSBlood samples from 123 schizophrenic patients with aggressive behaviors and 489 schizophrenic patients without aggressive behaviors were collected. DNA from all samples was amplified with a PowerPlex 21 system and separated by electrophoresis to determine the genotypes and allelic frequencies of 20 STR loci including D3S1368, D1S1656, D6S1043, D13S317, Penta E, D16S639, D18S51, D2S1338, CSF1PO, Penta D, TH01, vWA, D21S11, D7S820, D5S818, TPOX, D8S1179, D12S391, D19S433, and FGA.

RESULTSAll of the 20 STR loci have reached Hardy-Weinberg equilibrium in both groups. A significant difference was found in allelic and genotypic frequencies of loci Penta D between the two groups (alleles: P=0.042; genotypes: P=0.014) but not for the remaining 19 loci (P> 0.05). Univariate analysis also showed a significant difference for allele 10 and genotypes 10-12 of Penta D between the two groups (P=0.0027, P=0.0001), with the OR being 1.81 (95%CI: 1.22-2.67) and 4.33 (95%CI: 1.95-9.59), respectively.

CONCLUSIONPenta D may be associated with aggressive behaviors of schizophrenia. Allele 10 and genotypes 10-12 of Penta D may confer a risk for the disease.

Adolescent ; Adult ; Aged ; Aggression ; Gene Frequency ; Genotype ; Humans ; Microsatellite Repeats ; Middle Aged ; Schizophrenia ; genetics ; Young Adult

Objective To discuss the effect of DNA extraction of bloodstain on the filter paper with four methods of solid phase absorption.Methods 180 bloodstain samples on the iflter paper, each one contains 1 microlitre anticoagulation peripheral venous blood, divided into 4 groups with 45 samples, respectively. All samples were treated with four methods of solid phase absorption, i.e. DNA IQ? System, D-shield sensitive DNA Extraction Kit, High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method. The concentration of DNA and the results of STR typing of four groups were compared each other.Results The concentration of DNA was 3.764±1.790μg/mL and 3.634±1.112μg/mL by using D-shield sensitive DNA Extraction Kit and High efifciency Silica Bead DNA Extraction Kit, respectively. However, the concentration of DNA by using Conventional silica bead method group (3.350±1.250) was not signiifcantly different from each other (P<0.05), while the concentration of DNA extracted with above three methods were higher than by using DNA IQ? System (1.864±1.207)(P<0.001); Signiifcant differences of peak height existed between DNA IQ? System and other three methods (P<0.001); As the same time, the peak height of samples by using High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method were signiifcantly different from D-shield sensitive DNA Extraction Kit (P<0.01).Conclusions The DNA extracted in bloodstain on the iflter paper by using D-shield sensitive DNA Extraction Kit, High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method was more than DNA IQ? System. Meanwhile, the quality of DNA using High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method may be higher.

OBJECTIVETo assess the association between aggressive behaviors and 15 short tandem repeats (STRs) loci.

METHODSPeripheral blood samples from 541 army men with aggressive behaviors and 459 healthy individuals were collected. All sample were amplified with a AmpFlSTR Identifiler(TM) system and separated by electrophoresis to compare the genotypic and allelic frequencies of 15 STRs (CSF1PO, D2S1338, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, FGA, THO1, TPOX and vWA) in the two groups.

RESULTSA significant difference was found in allelic and genotypic frequencies at loci D2S1338 and D19S433 (P< 0.01) between the two groups, but not for the remaining 13 STR loci (P> 0.05). Univarite analysis also showed a significant difference for allele 16, genotypes 19-22, 22-24 on D2S1338 and genotypes 13-14.2 on D19S433 between the two groups (P= 0.0018, P= 0.0001, P= 0.0003, P= 0.0000), with the OR values being 7.380 (95%CI: 1.701-32.028), 0.051(95%CI: 0.007-0.388), 13.933(95%CI: 1.845-105.717), 0.349 (95%CI: 0.216-0.564), respectively.

CONCLUSIOND2S1338 and D19S433 may be associated with aggressive behavior. Allele 16 and genotype 22-24 on D2S1338 may be susceptible factors for the disease, whilst genotypes 19-22 on D2S1338 and 13-14.2 on D19S433 may confer a protective effect on it.

Adult ; Aggression ; Alleles ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Mental Disorders ; genetics ; Microsatellite Repeats

Objective To investigate the relationship of the initiative-aggressive behaviors and short tandem repeats (STRs) loci Penta D and Penta E.Methods The biological samples of 307 initiativeaggressive violent offenders and 459 healthy persons without violent behavior were collected.Then all of the sample were amplified by PowerPlex(R) 16 HS System and separated by electrophoresis to compare the genotypes and alleles frequency of Penta D and Penta E in both groups.Results Penta D and Penta E loci in both groups were found to coincide with Hardy-Weinberg equilibrium(P ＞0.05).There were significant difference of alleles and genotypes frequency at loci Penta D and Penta E between both groups (alleles:P =0.000 0,P =0.001 3; genotypes:P =0.001 2; P =0.000 0).Univarite analysis showed significant differences at allelle 5,6 on Penta D and allelle 9 on Penta E between both groups (P =0.002 4,P =0.001 6,P =0.002 1),with the OR values being 0.113 (95％ CI:0.015-0.869),12.092 (95 ％ CI:1.506-96.924),0.225 (95％ CI:0.078-0.648),respectively.Conclusion Penta D and Penta E might be associated with initiative-aggressive behavior,with allelle 6 on Penta D being the susceptible factor and allelle 5 on Penta D and allelle 9 on Penta E might being the protective factors.

Objective To investigate the relationship of the initiative-aggressive behaviors and short tandem repeats (STRs) loci Penta D and Penta E.Methods The biological samples of 307 initiativeaggressive violent offenders and 459 healthy persons without violent behavior were collected.Then all of the sample were amplified by PowerPlex(R) 16 HS System and separated by electrophoresis to compare the genotypes and alleles frequency of Penta D and Penta E in both groups.Results Penta D and Penta E loci in both groups were found to coincide with Hardy-Weinberg equilibrium(P ＞0.05).There were significant difference of alleles and genotypes frequency at loci Penta D and Penta E between both groups (alleles:P =0.000 0,P =0.001 3; genotypes:P =0.001 2; P =0.000 0).Univarite analysis showed significant differences at allelle 5,6 on Penta D and allelle 9 on Penta E between both groups (P =0.002 4,P =0.001 6,P =0.002 1),with the OR values being 0.113 (95％ CI:0.015-0.869),12.092 (95 ％ CI:1.506-96.924),0.225 (95％ CI:0.078-0.648),respectively.Conclusion Penta D and Penta E might be associated with initiative-aggressive behavior,with allelle 6 on Penta D being the susceptible factor and allelle 5 on Penta D and allelle 9 on Penta E might being the protective factors.