Demonstration of safety characteristics and effects on gut microbiota of Lactobacillus gasseri HMV18

Human normal flora is a source of probiotics.The safety characteristics of a specific isolate determine its application in foods or drugs.The food-borne-pathogen antagonist strain Lactobacillus gasseri HMV18 is one of the isolates from normal human flora.In this work,we assessed the in vitro pH tolerance,bile tolerance,biogenic amine production,mucin utilization,and safety of in vivo administration to mice to evaluate general health,organ-body weight index,organ histopathological change,whether L.gasseri HMV18 can colonize in the gut or modulate the gut microbiota after oral administration.The results suggest that L.gasseri HMV18 can tolerate pH 3 for 2 h,3%bile for 3 h,biogenic amine negative,mucin usage negative,does not encode verified toxins,and cause no visible change in mice's organs.L.gasseri HMV18 might not colonize in mice's gut,but can significantly affect the structure of gut microbiota.A bibliographical survey suggested that there were as few as 8 opportunistic infection cases from 1984 to 2022 and that the possibility for L.gasseri to cause infection is relatively low.Therefore,this work provides a basis for the foods or drugs application of L.gasseri HMV18 and gives a map of experiments for the safety assessment of probiotics.

Messenger RNA Profiling for Forensic Body Fluid Identification： Research and Applications

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts.However,the conventional body fluid identification methods are prone to various limitations,such as time consumption,intensive labor,nonparallel manner,varying degrees of sensitivity and limited specificity.Recently,the analysis of cell-specific messenger RNA expression(mRNA profiling)has been proposed to supplant conventional methods for body fluid identification.Since 2011,the collaborative exercises have been organized by the European DNA Profiling Group(EDNAP)in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification.The major advantages of mRNA profiling,compared to the conventional methods,include higher sensitivity,greater specificity,the ability of detecting several body fluids in one multiplex reaction,and compatibility with current DNA extraction and analysis procedure.In the current review,we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.

Assessment of a Sudden Death Case due to Coronary Artery Disease Based on the PMCT and Forensic Autopsy

It is never an easy thing to diagnose heart vascular disease only depending on the unenhanced postmortem computed tomography(PMCT).This article reported a case of sudden natural death after the complaint of anterior chest pain in which coronary artery calcification(CAC) was clearly displayed using PMCT scan.The entire coronary artery system was almost reconstructed via multiplanar reformation(MPR) and volume-rendering reconstruction(VR),and the total calcium score of the coronary arteries was obtained with CaScoring automatic analysis software.The results showed that CAC was conspicuous;the total calcium score was 640.3,considerably higher than 400.The pulmonary ground-glass opacity(GGO) and small amount of fluid both in the subglottic trachea and main bronchi were also found.The imaging results confirmed those of autopsy.In addition,the results concluded that PMCT might serve as an invaluable adjunct to the classic autopsy procedure.

Rapid screening of 420 pesticide residues in fruits and vegetables using ultra high performance liquid chromatography combined with quadrupole-time of flight mass spectrometry

A new method for screening and identification 420 pesticide residues in fruits and vegetables by ultra-performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry(UPLC-Q-TOF/MS)were developed.The samples were extracted with acetonitrile/acetic acid(99:1,V/V),and clean-up by SinChERS-Nano(single-step,cheap,effective,rugged,safe,nano)column,determined by UPLC-Q-TOF/MS.The accurate mass database and MS/MS database which contains 420 pesticides were established,the automatic retrieval of detection results was carried on according to the accurate mass,retention time,isotope ratio,ion fragment information,and so on.Method verification was performed on leeks samples.The results showed that 420 pesticides had good linearity in the range of 0.1-100μg/L,and the correlation coeffificients(R2)was greater than 0.990.The limits of detections(LODs)and limits of quantifications(LOQs)of 420 pesticides were in range of 0.05-2.0 and 0.1-5.0μg/L,respectively.The average spike recoveries at 3 levels were 70.1%to 119.7%,and the relative standard deviations(RSD)were lower than 20%(n=6).With this method,a survey of pesticide residues was conducted for 110 samples of 10 different fruits and vegetables,which provided scientific data for ensuring pesticide residue safety of the fruits and vegetables consumed daily by the public.This method was simple,sensitive and accurate,and could be used for rapid screening of 420 pesticide residues in fruits and vegetables.

Intermittent fasting boosts antitumor immunity by restricting CD11b^(+)Ly6C^(low)Ly6G^(low) cell viability through glucose metabolism in murine breast tumor model

Intermittent fasting can benefit breast cancer patients undergoing chemotherapy or immunotherapy.However,it is still uncertain how to select immunotherapy drugs to combine with intermittent fasting.Herein we observed that two cycles of fasting treatment significantly inhibited breast tumor growth and lung tissue metastasis,as well as prolonged overall survival in mice bearing 4T1 and 4T07 breast cancer.During this process,both the immunosuppressive monocytic-(M-)and granulocytic-(G-)myeloid-derived suppressor cell(MDSC)decreased,accompanied by an increase in interleukin(IL)7R^(+)and granzyme B^(+)T cells in the tumor microenvironment.Interestingly,we observed that Ly6G^(low)G-MDSC sharply decreased after fasting treatment,and the cell surface markers and protein mass spectrometry data showed potential therapeutic targets.Mechanistic investigation revealed that glucose metabolism restriction suppressed the splenic granulocytemonocyte progenitor and the generation of colony-stimulating factors and IL-6,which both contributed to the accumulation of G-MDSC.On the other hand,glucose metabolism restriction can directly induce the apoptosis of Ly6G^(low)G-MDSC,but not Ly6G^(high)subsets.In summary,these results suggest that glucose metabolism restriction induced by fasting treatment attenuates the immune-suppressive milieu and enhances the activation of CD3^(+)T cells,providing potential solutions for enhancing immune-based cancer interventions.

Reliability Study on Quantitative Detection of Extensor Digi- torum Brevis Strength with Needle Electromyography and Nerve Conduction

Objective To study the objectivity and reliability of needle electromyography and nerve conduction for detection of musculus extensor digitorum brevis strength, which may provide a basis for establishing a quantitative detection of muscle strength in forensic clinical study. Methods Forty-four healthy people were enrolled as the subjects, and during toe dorsiflexion, the following items including needle electromyography indexes, motor unit potential(MUP) amplitude, MUP count, recruitment reaction type, and nerve conduction detection indexes, compound muscle action potential(CMAP) amplitude, CMAP latent period and motor nerve conduction velocity(MNCV), were simultaneously detected under the cooperation and disguise condition. Results Under the cooperation condition, regardless of the same operator or different operators, there were good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, while there were normal test-retest reliabilities in MUP count and recruitment reaction type and the repeatability of the same operator was slightly better than the repeatability between different operators. Under the disguise condition, test-retest reliabilities of MUP amplitude, CMAP amplitude, CMAP latent period and MNCV were relatively high, while test-retest reliabilities of MUP count and recruitment reaction type were relatively low. Conclusion There are good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, which can be conducive to comparison between different operators and results at various times; MUP count and recruitment reaction type, which can be easily affected by subjectivity of operators and examinees, can be used to differentiate whether an examinee disguises or not. The indexes used to objectively judge muscle strength remain to be further investigated.

Rapid analysis of fifteen sulfonamide residues in pork and fish samples by automated on-line solid phase extraction coupled to liquid chromatography–tandem mass spectrometry

The aim of this work was to develop an automated on-line solid phase extraction(SPE)with liquid chromatography-tandem mass spectrometry method for the detection of fifteen sulfonamides in pork and fish samples.Samples were extracted with 0.2%formic acid acetonitrile solution,purified by on-line SPE device with HLB column,then separated by XBridge C18 column,using 0.1%formic acid solution and acetonitrile as the mobile phase.Mass spectrometric data was acquired under multiple reaction monitoring(MRM)mode using positive ionization electrospray.Internal standard method was used in the quantification,good linear relationship was got in range of 0.1–100 ng/mL and correlation coefficient was higher than 0.9990.The limits of detection were in the range of 0.125–2.00g/kg and the limits of quantitation were in the range of 0.250–5.00g/kg.Recoveries of the method were in range of 78.3%–99.3%,relative standard deviation were lower than 10%.The method was simple,sensitivity,and could be used for routine supervision and analysis of fifteen sulfonamides in pork and fish.

Simultaneous determination of 15 pesticide residues in Chinese cabbage and cucumber by liquid chromatography-tandem mass spectrometry utilizing online turbulent flow chromatography

In this experiment,a liquid chromatography tandem mass spectrometry method was built to determine 15 pesticide residues in Chinese cabbage and cucumber samples based on online turbulent flow chromatography purification.After modified quick,easy,cheap,effective,rugged,and safe(QuEChERS)extraction,extracts were directly injected to the TLX(TurboFlow Liquid Xcalibur)system and brought to TurboFlow™columns for on-line purification and then transferred to analytical column for further separation and analysis.TurboFlow™columns types,transfer flow rate,and transfer time were optimized.Limits of detection and limits of quantification of the method obtained for 15 pesticide residues were ranged between 0.2–1.0μg/kg and 0.5–2.0μg/kg in Chinese cabbage and cucumber samples.Recoveries of pesticide residues were in range of 75.3%–103.7%.Matrix effects for 15 pesticides were in range of 5.6%–106.6%.The developed method has been successfully used for the determination of 15 pesticide residues in real samples.

A new Lactobacillus gasseri strain HMV18 inhibits the growth of pathogenic bacteria

To search for a new eco-friendly therapy for infectious disease caused by Escherichia coli,Staphylococcus aureus or Klebsiella oxytoca,we collected the vaginal swabs from healthy women,screened for Lactobacillus and found a strain repressing the growth of pathogenic bacteria.The new isolate was identified as L.gasseri by the colony morphology,Gram staining,biochemical reactions and confirmed by the 16 S rDNA sequencing.The HMV18 strain inhibited the growth of food-borne pathogens such as E.coli,S.aureus and K.oxytoca.The HMV18 strain was sensitive to penicillin,ampicillin,erythromycin,tetracycline and chloramphenicol.The HMV18 strain producedα-hemolysis.Pathological histology of the mice ileum showed that the mucosa,villi,lamina propria and crypt depth remained intact and there was no inflammation or hyperemia in the L.gasseri HMV18 gavaged group.L.gasseri HMV18 could not up-regulate inflammatory cytokines level of plasma.All the results suggested L.gasseri HMV18 is a candidate probiotic to be an additive for food preservation or drug to prevent food-borne diseases.

Severe Hypokalemia due to Combination Use of Dexamethasone and Calcium Gluconate for Diabetics with Infection： A Case Report

The combination use of dexamethasone and calcium gluconate can be applied to hypersensitivity. Severe hypokalemia is a usual complication of dexamethasone and calcium gluconate therapy, which occurs frequently with therapeutic use. Fatal cases, accidental and intentional, occur frequently in forensic practice. The current case report presented a 43-year-old man with diabetes mellitus with infection, to whom dexamethasone and calcium gluconate were administered in the private clinic. With the development of such clinical symptoms of severe hypokalemia as quadriplegia, he was confirmed to have severe hypokalemia through a biochemical test before dying of arrhythmia. And also it presented pathophysiologic mechanism underlying severe hypokalemia as well as suggestions for clinical practice regarding combination use of dexamethasone and calcium gluconate.

Genome and transcriptome of Papaver somniferum Chinese landrace CHM indicates that massive genome expansion contributes to high benzylisoquinoline alkaloid biosynthesis

Opium poppy(Papaver somniferum)is a source of morphine,codeine,and semisynthetic derivatives,including oxycodone and naltrexone.Here,we report the de novo assembly and genomic analysis of P.somniferum traditional landrace‘Chinese Herbal Medicine’.Variations between the 2.62 Gb CHM genome and that of the previously sequenced high noscapine 1(HN1)variety were also explored.Among 79,668 protein-coding genes,we functionally annotated 88.9%,compared to 68.8%reported in the HN1 genome.Gene family and 4DTv comparative analyses with three other Papaveraceae species revealed that opium poppy underwent two whole-genome duplication(WGD)events.The first of these,in ancestral Ranunculales,expanded gene families related to characteristic secondary metabolite production and disease resistance.The more recent species-specific WGD mediated by transposable elements resulted in massive genome expansion.Genes carrying structural variations and large-effect variants associated with agronomically different phenotypes between CHM and HN1 that were identified through our transcriptomic comparison of multiple organs and developmental stages can enable the development of new varieties.These genomic and transcriptomic analyses will provide a valuable resource that informs future basic and agricultural studies of the opium poppy.

Electroretinogram and Histopathologic Changes of the Retina after Methanol Intoxication

In order to study the functional and structural alterations of the retina in SD rat model after methanol intoxication,35 rats were divided randomly into five groups administrated with saline,3-day high dose,7-day high dose,3-day low dose and 7-day low dose methanol separately.The retinal function of each group was assessed by flash electroretinogram(F-ERG) 3 and 7 days after methanol poisoning.The microstructure and ultrastructure of the retina were observed at the same time.The high-dose methanol intoxication induced irreversible retinal functional and structural damages 3 days after poisoning,which included prolonged latency and reduced amplitude of the Max-reaction of F-ERG.These injuries were aggravated 7 days after poisoning.Meanwhile,the latency and amplitude of the Cone-reaction of F-ERG were also affected 3 days after poisoning,but there were no further worsening tendency 7 days after poisoning.The retinal histological analysis showed cellular edema,heteromorphy and disarrangement,tissular loosen of the inner nuclear layer and photoreceptors layer.The mitochondrial damage began at the photoreceptors layer and developed further into the inner nuclear layer.The low-dose methanol intoxication only caused transient damage of the retina.Our results showed that the function and structure of the photoreceptor and inner nuclear layer were the primary target of methanol intoxication and that the rod cells were more sensitive to methanol intoxication than the cone cells.The mitochondrial damage developed from outer layer to inner layer of the retina.

Stature Estimation Based on the Length of Tibia and Fibula Measured by Digital X-ray in Chinese Han Teenagers

Objective To derive regression formulae for stature estimation using forensic radiography from the tibia and fibula of Chinese Han teenagers in Sichuan Province. Methods To construct equations, measurements were conducted on the training sample (412 adults, 201 males and 211 females). The whole length of the fibula and four measurements of tibia were determined using CR radiography, rectified through theoretical magnification. The regression formulae were relatively constructed to the real stature measured in an erect position. Through using the testing sample (40 adults) for the regression formulae, the reliability of the regression formulae was assessed. Results The range of correlation coefficients of four measurements for tibia was 0.880-0.895 in the sex-unknown group, 0.869-0.893 in the male, and 0.8450.855 in the female. The five measurements were found to be better correlated with stature in the male than in the female. Conclusion The digital X-ray of the tibia and fibula for stature estimation is proved to be effective in forensic individual identification; therefore, these equations can be of great assistance to the stature estimation of the contemporary Chinese Han teenagers.

Long-Term Trend of Bone Development in the Contemporary Teenagers of Chinese Han Nationality

Objective To further improve the accuracy of bone age identification using the time of secondary ossification center appearance and epiphyseal fusion of 7 joints to estimate the age of living individuals.Methods DR films were taken from 7 parts including sternal end of clavical and the left side of shoulder,elbow,carpal,hip,knee and ankle joints of 1 709 individuals who came from eastern China,central China and southern China,whose ages were between 11.0 and 20.0 years.From those 7 joints 24 osteal loci were selected as bone age indexes,which could better reflect age growth of teenagers.The characteristics of secondary ossification center appearance and epiphyseal fusion were observed,and the mean and age range of secondary ossification center appearance and epiphyseal fusion were calculated.Results The fusion time of the 24 epiphyses were advanced at different degrees,the most obvious epiphyses the sternal end of clavicle,scapular acromial end,distal end of the radius,distal end of the ulna,iliac crest,ischial tuberosity,the upper and lower end of tibia and fibula.The appearance time of sternal end of clavicle,scapular acromial end,iliac crest and ischial tuberosity epiphyses were all found to be after the age of 12,and the female's age,approximately 1 year ahead of schedule in comparison with the male's.Conclusion The relevant forensic information and data for bone age identification should be updated every 10-15 years so as to provide accurate and objective evidence for court testimony,conviction and sentencing.

Postmortem MSCT Diagnosis of Whiplash Injuries in a Traffic Accident：A Case Report and Review of the Literature

A 45-year-old male car driver died in a traffic accident of four cars rear-end collision on the highway.He was found to have died after a respiratory and cardiac arrest at the scene.No sign of skin injuries was observed from the external inspection.The autopsy was not permitted by the family members because of the local culture.Multislice computed tomography(MSCT)was applied to the current case,showing dislocation of C3~4cervical vertebrae withⅡdegree,C4vertebral plate fractures,and spinal stenosis.Post-mortem MSCT confirmed the diagnosis as whiplash injuries.MSCT was verified to be effective in showing the severity of whiplash injuries,thus providing certain objective evidence for medicolegal expertise.

3D Finite Element Model of Human Knee Injuries in the Traffic Accident

Objective To explore the injury mechanism of the human knee in a traffic accident by establishing a 3D finite element(FE) model. Methods The FE model, composed of femur, tibia, fibula, patella, meniscus, knee ligaments and surrounding soft tissues, was reconstructed by CT scanning data from a male volunteer. Validation was performed by the lateral impact simulation, and the stress and strain results were obtained to be compared with those previously reported for injury prediction. Results The results derived from the FE model were found to be similar with those previously reported, most of the ligaments and meniscus wounded at 40 m/s collision, which was readily observed. Conclusion The simulation results generated by FE model can be effectively used for the injury mechanism analysis of initial contact.

Circular RNA expression alterations are involved in incubation of morphine craving

OBJECTIVE Cue-induced drug craving progressively increase after prolong abstinence in animal and human beings.A behavioral phenomenon termed incubation of drug craving is considered as a key reason for drug relapse,but the transcriptional mechanisms that contribute to this incubation are unknown.It has been demonstrated that circular RNAs(circR NAs),act as miR NA sponges,play important roles in the regulation of gene expression and the pathogenesis of disease.The present study aims to explore the transcriptional profiles associated with incubation of morphine craving in nucleus accumbens(NAc),an important brain area previously implicated in drug seeking.METHODS The animal model of the incubation of drug craving was induced by CPP paradigms with six morphine(5 mg·kg-1) injections and 14 d drug abstinence in mice.The brain tissues of NAc were collected after the behavioral tests for circRNA-sequencing by Illumina Hiseq X sequencer.We identified differentially expressed genes(DEGs) by qRT-PCR and used bioinformatics methods for further function analysis.RESULTS The progressive increase of CPP scores during the 14 d drug abstinence indicated the establishment of animal model.The data of circRNA-sequencing reported that 16 circRNAs were significantly altered after 28 d drug abstinence in NAc of morphine treated mice(FC≥2 and P<0.05).Among those circRNAs,9 were significantly up-regulated,while 7 were down-regulated.Furthermore,we subsequently tested circRNAs expression using quantitative real-time PCR,and the consistent data were obtained.The results of KEGG pathway analysis indicated that these genes were enriched for several biological processes,including RNA transport,protein ubiquitination and histone methylation,etc.CONCLUSION These findings provide a unique resource of gene expression data for future studies examining transcriptional mechanisms in NAc that mediate opioids seeking after prolonged withdrawal.

Genetic polymorphism of mitochondrial DNA HVS-I and HVS-II of Chinese Tu ethnic minority group

We analyzed the two hypervariable segments HVS-I and HVS-II of 108 Chinese Tu ethnic minority group samples for forensic and population genetics purposes, Comparing with Anderson sequence, 79 polymorphic loci in HVS-I and 40 in HVS-II were found in Chinese Tu ethnic minority group mtDNA sequences, and 90 and 64 haplotypes were then defined. Haplotype diversity and the mean pairwise differences were 0.9903±0.0013 and 5.7785 in HVS-I, and 0.9777±0.0013 and 3.5819 in HVS-II, respectively. By analyzing the hypervariable domain from nucleotide 1,6180 to 1,6193 in HVS-I, we defined some new types of sequence variations. We also compared the relationship between Tu population and other populations using mtDNA HVS-I sequences. According to Rst genetic distances, the phylogenetic tree showed that the Tu population, the Xi＇an Han population, the Chinese Korean, and the Mongol ethnic group were in a clade. This indicated a close genetic relationship between them. There were far relations between the Tu population and other Chinese southern Han populations, Siberian, European, African, and other foreign populations. The results suggest that Tu population has a multi-origin and has also merged with other local populations.

Mathematical Models for Teenager's Living Age Evaluation Based on CT Image of Medial Clavicular Epiphysis

Objective To explore the correlation between volume rendering(VR) statistics of medial clavicular epiphysis and living age,and establish the mathematical models for living age evaluation using the CT image of medial clavicular epiphysis based on the growth rules of osteoepiphysis of medial clavicle.Methods The CT images of the medial clavicles from 795 teenagers aged 15-25,387 males and 408 females,were collected in East and South China.VR 3D images were reconstructed from 0.60 mm-thick slice CT images.The epiphyseal diameter,sternal end diameter,and their respective diameter ratio(the left: x 1;the right: x 3);epiphyseal area,sternal end area,and their respective area ratio(the left: x 2;the right: x 4),were measured and calculated.All these observations were analyzed using SPSS 19.0 statistical software.The statistical differences in gender and age were analyzed by Mann-Whitney U test.The mathematical models were established using least square.Sixty trained subjects,30 males and 30 females,were tested to verify the accuracy of the established mathematical models.Results In the group of same age,x 1 showed significant difference in gender;the same results were observed in x 2,x 3,and x 4,which suggested that the growth rules of osteoepiphysis of medial clavicle were highly correlated with living age.The accuracy of these mathematical models were all above 67.6%(±1.0 year) and 78.5%(±1.5 year).Conclusion The mathematical models with reasonable accuracy could be manageable in practice to confirm the conclusion of the atlas method.The current study can contribute to the single skeletal age evaluation.

Quality Control System of Judicial Expertise in China

The current quality control system of judicial expertise in China For a long time in China,the administrative organization of justice has been subordinate to universities,Ministry of Public Health,Ministry of Public Security,the Procuratorate,the Court and National Security Agency.These administrative organizations are mainly in charge of the management of judicial expertise.The quality control system of judicial expertise can be classified into three levels.