The Evolution, Expression Patterns, and Domestication Selection Analysis of the Annexin Gene Family in the Barley Pan-Genome.

Plant annexins constitute a conserved protein family that plays crucial roles in regulating plant growth and development, as well as in responses to both biotic and abiotic stresses. In this study, a total of 144 annexin genes were identified in the barley pan-genome, comprising 12 reference genomes, including cultivated barley, landraces, and wild barley. Their chromosomal locations, physical-chemical characteristics, gene structures, conserved domains, and subcellular localizations were systematically analyzed to reveal the certain differences between wild and cultivated populations. Through a cis-acting element analysis, co-expression network, and large-scale transcriptome analysis, their involvement in growth, development, and responses to various stressors was highlighted. It is worth noting that HvMOREXann5 is only expressed in pistils and anthers, indicating its crucial role in reproductive development. Based on the resequencing data from 282 barley accessions worldwide, genetic variations in thefamily were investigated, and the results showed that 5 out of the 12 identified HvMOREXanns were affected by selection pressure. Genetic diversity and haplotype frequency showed notable reductions between wild and domesticated barley, suggesting that a genetic bottleneck occurred on the annexin family during the barley domestication process. Finally, qRT-PCR analysis confirmed the up-regulation of HvMOREXann7 under drought stress, along with significant differences between wild accessions and varieties. This study provides some insights into the genome organization and genetic characteristics of the annexin gene family in barley at the pan-genome level, which will contribute to better understanding its evolution and function in barley and other crops.

Establishment and Application of a 42-plex Microhaplotype Assay in Forensic Genetics.

OBJECTIVES: To establish and forensically verify a 42 microhaplotypes (mircohaps, MHs) multiplex assay system based on next-generation sequencing (NGS), and to explore the application value of this system in the practice of forensic genetics. METHODS: A total of 42 highly polymorphic MHs were selected from previous studies, and sequenced by the MiSeq FGxTM platform to verify the repeata-bility, sensitivity, specificity, stability, and mixture analysis ability of the detection system. Through population genetic investigation of 102 unrelated Chinese Han individuals in Liyang City, Jiangsu Province, China, the application value of this system in forensic genetics was evaluated. RESULTS: The sequencing repeatability of the 42-plex MHs assay was 100% and the sensitivity was as low as 0.062 5 ng. The system had the ability to withstand the interference of indigo (≤2 500 ng/µL), humic acid (≤9 ng/µL), hemoglobin(≤20 µmol), and urea (≤200 ng/µL) and to detect mixtures of 2 people (1∶19), 3 people (1∶1∶9) and 4 people (1∶1∶1∶9). Based on 102 individual data, the combined power of discrimination and the combined power of exclusion were 1-3.45×10-30 and 1-3.77×10-11, respectively, and the average effect value of alleles was 2.899. CONCLUSIONS: The 42-plex MHs assay was successfully established in this study and this system has high repeatability and sensitivity, good anti-jamming ability and mixture analysis ability. The 42 MHs are highly polymorphism and have good application value in individual identification and paternity testing.

A diagnostic strategy for pulmonary fat embolism based on routine H&E staining using computational pathology.

Pulmonary fat embolism (PFE) as a cause of death often occurs in trauma cases such as fractures and soft tissue contusions. Traditional PFE diagnosis relies on subjective methods and special stains like oil red O. This study utilizes computational pathology, combining digital pathology and deep learning algorithms, to precisely quantify fat emboli in whole slide images using conventional hematoxylin-eosin (H&E) staining. The results demonstrate deep learning's ability to identify fat droplet morphology in lung microvessels, achieving an area under the receiver operating characteristic (ROC) curve (AUC) of 0.98. The AI-quantified fat globules generally matched the Falzi scoring system with oil red O staining. The relative quantity of fat emboli against lung area was calculated by the algorithm, determining a diagnostic threshold of 8.275% for fatal PFE. A diagnostic strategy based on this threshold achieved a high AUC of 0.984, similar to manual identification with special stains but surpassing H&E staining. This demonstrates computational pathology's potential as an affordable, rapid, and precise method for fatal PFE diagnosis in forensic practice.

[mRNA Expression Profile Changes in Angiotensin-â ¡-Induced Atrial Myocardial Fibrosis in Rats].

Objective: To study the differences between the mRNA expression profile in angiotensin Ⅱ (Ang Ⅱ)-induced fibrotic cardiomyocytes and that of normal cardiomyocytes and the relevant signaling pathways. Methods: Six 8-week-old male Sprague-Dawley (SD) rats were randomly assigned to a control group and an Ang Ⅱ group, with 3 rats in each group. Rats in the control group were injected via caudal vein with 0.9% normal saline at 2 mg/kg per day, while rats in the Ang Ⅱ group were injected with Ang Ⅱ via caudal vein at 2 mg/kg per day. The medications were continuously administered in the two groups for 14 days. The degree of myocardial fibrosis was determined by Masson's Trichrome staining and the content of collagen Ⅰ was determined by immunohistochemistry. High throughput sequencing was performed to measure the mRNA expression of rat cardiomyocytes in the two groups and to screen for differentially-expressed mRNAs. The differentially-expressed mRNAs were analyzed by Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Results: Compared with those of the control group, the degree of myocardial fibrosis and the content of collagen Ⅰ in Ang Ⅱ group were significantly higher ( P<0.05). Through sequencing, 313 differentially-expressed mRNAs were identified, with 201 being up-regulated and 112 being down-regulated. Go and KEGG analyses showed that these differentially-expressed mRNA were involved in a variety of biological regulatory functions and pathways of myocardial fibrosis. Conclusion: Ang Ⅱ can cause myocardial fibrosis in rats. There are significant differences in mRNA expression between fibrotic cardiomyocytes and normal cardiomyocytes. The differentially expressed mRNAs may play an important role in biological processes, including immune response, cell remodeling, and extracellular matrix deposition.

Establishing an hTERT-driven immortalized umbilical cord-derived mesenchymal stem cell line and its therapeutic application in mice with liver failure.

Acute liver failure (ALF) is characterized by rapid liver cell destruction. It is a multi-etiological and fulminant complication with a clinical mortality of over 80%. Therapy using mesenchymal stem cells (MSCs) or MSCs-derived exosomes can alleviate acute liver injury, which has been demonstrated in animal experiments and clinical application. However, similar to other stem cells, different cell sources, poor stability, cell senescence and other factors limit the clinical application of MSCs. To achieve mass production and quality control on stem cells and their exosomes, transfecting umbilical cord mesenchymal stem cell (UCMSC) with lentivirus overexpressing human telomerase reverse transcriptase (hTERT) gene, the hTERT-UCMSC was constructed as an immortalized MSC cell line. Compared with the primary UCMSC (P3) and immortalized cell line hTERT-UCMSC at early passage (P10), the hTERT-UCMSC retained the key morphological and physiological characteristics of UCMSC at the 35th passage (P35), and showed no signs of carcinogenicity and toxic effect in mice. There was no difference in either exosome production or characteristics of exosomes among cultures from P3 primary cells, P10 and P35 immortalized hTERT-UCMSCs. Inoculation of either hTERT-UCMSC (P35) or its exosomes improved the survival rate and liver function of ALF mice induced by thioacetamide (TAA). Our findings suggest that this immortalized cell line can maintain its characteristics in long-term culture. Inoculation of hTERT-UCMSC and its exosomes could potentially be used in clinics for the treatment of liver failure in the future.

Internal validation of an improved system for forensic application: a 41-plex Y-STR panel.

Y-chromosome short tandem repeats (Y-STRs) have a unique role in forensic investigation. However, low-medium mutating Y-STRs cannot meet the requirements for male lineage differentiation in inbred populations, whereas rapidly mutating (RM) high-resolution Y-STRs might cause unexpected exclusion of paternal lineages. Thus, combining Y-STRs with low and high mutation rates helps to distinguish male individuals and lineages in family screening and analysis of genetic relationships. In this study, a novel 6-dye, 41-plex Y-STR panel was developed and validated, which included 17 loci from the Yfiler kit, nine RM Y-STR loci, 15 low-medium mutating Y-STR loci, and three Y-InDels. Developmental validation was performed for this panel, including size precision testing, stutter analysis, species specificity analysis, male specificity testing, sensitivity testing, concordance evaluation, polymerase chain reaction inhibitors analysis, and DNA mixture examination. The results demonstrated that the novel 41-plex Y-STR panel, developed in-house, was time efficient, accurate, and reliable. It showed good adaptability to directly amplify a variety of case-type samples. Furthermore, adding multiple Y-STR loci significantly improved the system's ability to distinguish related males, making it highly informative for forensic applications. In addition, the data obtained were compatible with the widely used Y-STR kits, facilitating the search and construction of population databases. Moreover, the addition of Y-Indels with short amplicons improves the analyses of degraded samples. Key Points: A novel multiplex comprising 41 Y-STR and 3 Y-InDel was developed for forensic application.The multiplex included rapidly mutating Y-STRs and low-medium mutating Y-STRs, which is compatible with many commonly used Y-STR kits.The multiplex is a powerful tool for distinguishing related males, familial searching, and constructing DNA databases.

Epidemiological characteristics and secular trend of the HIV/AIDS cases among 15-24 years old population in Hefei from 2004 to 2022 / 中国学校卫生

Objective@#To investigate the characteristics of HIV/AIDS cases among 15-24 year old population reported in Hefei from 2004 to 2022, so as to provide insights into AIDS control among adolescents.@*Methods@#The epidemiological data regarding HIV/AIDS cases between 15 and 24 years old reported in Hefei from 2004 to 2022 were captured from the AIDS comprehensive prevention and control information system of Chinese disease prevention and control information system, and data regarding temporal distribution, population distribution, and routes of infections and detection were descriptively analyzed by descriptive epidemiological methods.@*Results@#From 2004 to 2022, 865 cases of HIV/AIDS were reported in Hefei among 15-24 years old youth, accounting for 21.80% of the total reported cases. Among the HIV/AIDS patients, males accounted for 92.60%(801 cases), the unmarried ones accounted for 93.41% (808 cases)，those with college degree or above accounted for 60.12% (520 cases)，and 25.78%(223 cases) of them were students. The proportion of student cases increased annually( χ 2 trends =47.67, P <0.01). Homosexual transmission accounted for 81.39%, both showed an increasing trend( χ 2 trends =51.23, P <0.01)．Totally 55.49% of cases were found through testing and consultation, and the proportion of cases increased by year( χ 2 trends =112.18, P <0.01). In 2004-2022，the number of newly reported cases among people aged 15-24 showed a rising trend at an average rate of 24.46% by year( Z=4.92, P <0.01), which was higher than the average rate of 21.54% for the entire population( Z=12.75, P <0.01).@*Conclusion@#The epidemic of HIV/AIDS among population aged 15-24 years is serious in Hefei. Comprehensive measures for HIV education and prevention intervention are desperately needed to be reinforced among targeted students.

Electrospun Polycrown Ether Composite Nanofibers as an Adsorbent for On-Line Solid Phase Extraction of Eight Bisphenols from Drinking Water Samples with Column-Switching Prior to High Performance Liquid Chromatography.

Bisphenols (BPs) are a class of endocrine disruptors widely existing in the environment. They have a great impact on human health owing to their environmental endocrine disrupting effects, chronic toxicity, neurotoxicity, cytotoxicity and genetic toxicity. In this paper, an on-line packed fiber solid phase extraction (PFSPE) coupling with column-switching HPLC-FLD determination method was developed for the determination of eight BPs in drinking water. The poly (dibenzo-18-crown-6-ether)/polystyrene composite nanofibers (PDB18C6/PS) were prepared by electrospinning and used as an adsorbent for the on-line PFSPE column. The on-line PFSPE-HPLC equipment contained a dual ternary pump and a switching valve to enable enrichment, purification, and analysis directly in the system. The results showed that the proposed on-line PFSPE-HPLC-FLD method realized the simultaneous separation and detection of eight BPs: BPF, BPE, BPA, BPB, BPAF, BPAP, BPC and BPZ. The curves of the target analytes were prepared with good correlation coefficient values (r2 > 0.998) in the range of 50−1000 pg/mL. The limit of detection (S/N = 3) was 20 pg/mL, the limit of quantitation (S/N = 10) is 50 pg/mL. The recoveries of eight BPs were 94.8−127.3%, and the intra-day precisions (RSD) were less than 10%. The PFSPE column made of the PDB18C6/PS composite nanofibers has stable properties and can be reused at least 200 times. In the detection of drinking water samples, BPZ was detected in nearly 80% of drinking water samples, and BPA, BPAP, BPF and BPAF were also detected in some water samples. This high level of integration and automation was achieved in pretreatment of eight BPs from water samples. The proposed simple, rapid, and practical method has been successfully applied to the detection of eight BPs in drinking water, which can provide powerful technical support for drinking water quality and safety monitoring.

Genetic Polymorphism of 16 X-STR Loci in Xinjiang Uygur Population.

OBJECTIVES: To study the genetic polymorphism and population genetic parameters of 16 X-STR loci in Xinjiang Uygur population. METHODS: The Goldeneye® DNA identification system 17X was used to amplify 16 X-STR loci in 502 unrelated individuals (251 females and 251 males). The amplified products were detected by 3130xl genetic analyzer. Allele frequencies and population genetic parameters were analyzed statistically. The genetic distances between Uygur and other 8 populations were calculated. Multidimensional scaling and phylogenetic tree were constructed based on genetic distance. RESULTS: In the 16 X-STR loci, a total of 67 alleles were detected in 502 Xinjiang Uygur unrelated individuals. The allele frequencies ranged from 0.001 3 to 0.572 4. PIC ranged from 0.568 8 to 0.855 3. The cumulative discrimination power in females and males were 0.999 999 999 999 999 and 0.999 999 999 743 071, respectively. The cumulative mean paternity exclusion chance in trios and in duos were 0.999 999 997 791 859 and 0.999 998 989 000 730, respectively. The genetic distance between Uygur population and Kazakh population was closer, and the genetic distance between Uygur and Han population was farther. CONCLUSIONS: The 16 X-STR loci are highly polymorphic and suitable for identification in Uygur population, which can provide a powerful supplement for the study of individual identification, paternity identification and population genetics.

Composite Nanofibers as Novel Sorbents for On-Line and Off-Line Solid-Phase Extraction in Chromatographic System: A Comparison for Detection of Free Biogenic Monoamines and Their Metabolites in Plasma.

Two different pretreatment approaches have been used for the enrichment and separation of biogenic monoamines and metabolites in plasma for high performance liquid chromatography (HPLC) determination. The first approach, based on on-line packed-fiber solid-phase extraction (PFSPE) coupled with HPLC, allows for the simultaneous detection of epinephrine (E), norepinephrine (NE), dopamine (DA), 3-methoxyl epinephrine (MN), norepinephrine (NMN), 3-methoxytyramine (3-MT), and 5-hydroxytryptamin (5-HT). Using this developed on-line PFSPE-HPLC method, the limit of detections (LODs) of the seven analytes ranged from 1 ng/mL (NMN and MN) to 2 ng/mL (NE, E, DA, 3-MT and 5-HT). The reportable ranges were 5-300 ng/mL for NE and DA, 5-100 ng/mL for E, and 5-200 ng/mL for NMN, MN, 3-MT and 5-HT. The off-line PFSPE-HPLC was employed in the second approach and could provide simultaneous detection of NE, E, DA, NMN, and MN. The linearity was verified in the range of 0.5-20 ng/mL (NE, E, and DA) and 20-250 ng/mL (NMN and MN). The LODs of the five analytes ranged from 0.2 ng/mL (NE, E, and DA) to 5 ng/mL (NMN and MN). This study verified the possibility of using nanofibers as an adsorbent in an on-line PFSPE-HPLC system for the determination of biogenic monoamines and their metabolites in human plasma. Compared with the off-line PFSPE approach, the on-line PFSPE method deserves attention mainly due to its greener character, derived from the automation of the process and high-throughput with less operators' handling.

Efficiency Analysis of Uncle-Nephew Relationship Identification by Increasing STR Markers and Adding Reference Samples.

OBJECTIVES: To estimate the system efficiency of uncle-nephew relationship identification by increasing STR markers and adding reference samples based on the test results of simulated data and real samples, so as to provide references for selecting the appropriate number of STRs and reference samples for uncle-nephew relationship identification. METHODS: Five common models of uncle-nephew relationship identification were constructed by adding different reference samples. In each model, the likelihood ratio (LR) for 10 000 pairs of uncle-nephew relationships and 10 000 pairs of unrelated individuals were simulated by detecting 19, 39 or 55 STRs, and the system efficiency at different thresholds was simulated. The samples of the Han population in Zhejiang were collected, and 55 autosomal STRs were obtained by using SiFaSTRTM 23plex kit, Goldeneye® DNA ID 22NC kit and AGCU 21+1 PCR amplification kit. When 19, 39 and 55 STRs were detected, the LR of each model and system efficiency under different thresholds were calculated and compared with the simulation results. RESULTS: Under the same detection system, the calculated results of simulated data and corresponding true samples were basically consistent. In the same model, there was a positive correlation between the system efficiency of uncle-nephew relationship identification and the number of STRs detected. Moreover, the system efficiency of introducing relatives was higher than identifying only two individuals. The order of preference for the introduction of relatives was the full sibling (or mother) of the uncle and the full sibling (or mother) of the nephew. CONCLUSIONS: The system efficiency of uncle-nephew relationship identification could be improved by increasing the number of STRs and introducing known relatives, which would provide the basis for selecting the most appropriate detection system and reference individuals in actual cases.

Molecular Characterization of Staphylococci Recovered from Hospital Personnel and Frequently Touched Surfaces in Tianjin, China.

Staphylococci are major hospital-associated pathogens, and the dissemination of methicillin-resistant staphylococci in hospitals poses a great challenge for managing hospital-acquired infections. Little is known about the dissemination of staphylococci recovered from the hospital environment and personnel in China. In this study, antimicrobial susceptibility tests, mecA gene detection, SCCmec typing, and multilocus sequence typing (MLST) were performed to clarify the molecular epidemiology of staphylococci in a large hospital in Tianjin, China. One hundred and ninety-five staphylococci were recovered, and 94% of isolates were resistant to at least one antibiotic. Eighty-five staphylococci were mecA gene-positive, and 40% of them harbored SCCmec IV and V. The genotype of Staphylococcus aureus (S. aureus) was ST25, and the dominant genotype of methicillin-resistant Staphylococcus epidermidis (MRSE) was ST59. Three new sequence types were assigned as ST840, ST841, and ST842. One (2%) frequently touched surface was contaminated by S. aureus, which suggested that environmental contamination occurred in the hospital in China. Nineteen (39%) frequently touched surfaces were contaminated by methicillin-resistant coagulase-negative staphylococci (MRCoNS), and 46% of HP carried MRCoNS. Varied staphylococcal species and antimicrobial-resistance rates were observed between staphylococci that were recovered from hospital personnel and frequently touched surfaces. The transmission of MRSE and S. aureus between hospital personnel and frequently touched surfaces was detected. Hospital items and personnel may act as reservoirs of antimicrobial-resistant staphylococci, and cleaning strategies should be carried out to decrease the dissemination of antimicrobial-resistant staphylococci in hospitals in China.

Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococci Recovered from Public Shared Bicycles in China.

Millions of public shared bicycles (PSBs) have been launched in China, and PSBs are a potential reservoir of antimicrobial-resistant staphylococci. However, no national data to elucidate the dissemination, antimicrobial resistance and genotypes of staphylococci has been recovered from public shared bicycles located in different cities in China. Antimicrobial susceptibility, SCCmec types and sequence types of staphylococci were determined. A total of 146 staphylococci were recovered in this study, and 87% staphylococcal isolates were resistant to at least one antibiotic. In total, 29 (20%) staphylococcal isolates harbored mecA gene, and SCCmec types were determined as follows: SCCmec type II (n = 1), IV(n = 3), V (n = 4), VI (n = 1), VIII (n = 2), A/1 (n = 6), A/5 (n = 2), C/1 (n = 2), C/2 (n = 1), C/3 (n = 1), (n = 5) and Pseudo (ψ)-SCCmec (n = 1). Sequence types of 16 Staphylococcus epidermidis were determined, including ST10, ST17, ST59, ST60, ST65, ST130, ST184, ST262, ST283, ST337, ST360, ST454, ST567, ST820, ST878 and ST934. PSBs are a reservoir of diverse antimicrobial-resistant staphylococci, and staphylococcal species differences were observed in isolates that were recovered from public shared bicycles in the south and north of China. PSBs are a source of antimicrobial resistance and genetic diverse staphylococci.

eQTL Highlights the Potential Role of Negative Control of Innate Immunity in Kawasaki Disease.

PURPOSE: Kawasaki disease (KD) is an acute systemic vasculitis mainly found in the medium-sized arteries, especially the coronary arteries. Immune system is involved in the pathogenesis of acute KD in children, but the functional differences in the immune system between healthy children and KD patients remain unclear. PATIENTS AND METHODS: A total of 190 KD patients and 119 healthy controls were recruited for the next-generation sequencing of 512 targeted genes from 4 immune-related pathways. Subsequently, the peripheral blood mononuclear cells (PBMCs) were isolated. RNA sequencing of the LPS treated PBMCs from additional 20 KD patients and 20 healthy controls was used to examine the differentially expressed genes (DEGs). Then, an expression quantitative trait locus (eQTL) analysis combined with previously analyzed RNA data were used to examine the DEGs. Finally, the serum levels of 13 cytokines were detected before and after LPS treatment in 40 samples to confirm the findings from eQTL analysis. RESULTS: A total of 319 significant eQTL were found, and both eQTL analysis and RNA sequencing showed some DEGs were involved in the connective tissue disorders and inflammatory diseases. DEGs that function to negatively regulate immunity were closely related to the pathogenesis of KD. In addition, the serum levels of IL-10 (an inflammatory and immunosuppressive factor) and SCD25 (an important immunosuppressant) reduced significantly in the KD patients. CONCLUSION: Our study shows the expression of factors responsible for the negative control of innate immunity is altered, which plays an important role in the etiology of KD.

Constructing a 3D interconnected "trap-zap" ß-CDPs/Fe-g-C3N4 catalyst for efficient sulfamethoxazole degradation via peroxymonosulfate activation: Performance, mechanism, intermediates and toxicity.

A novel and high-efficiency catalyst Fe doped g-C3N4 (Fe-g-C3N4) composited with ß-cyclodextrin polymers (ß-CDPs) was synthesized for activating peroxymonosulfate (PMS). The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) results showed that the catalyst was 3D interconnected porous structure. The degradation rate constant of sulfamethoxazole (SMX) in ß-CDPs/Fe-g-C3N4+PMS system was estimated to be 0.132 min-1, which was 14.7 times and 2.2 times that of g-C3N4+PMS and Fe-g-C3N4+PMS system, respectively. In addition, the ß-CDPs/Fe-g-C3N4 exhibited superior degradation performance in a wide pH range (3.0-9.0) and good selectivity in the presence of other inorganic anions and natural organics. Radical scavenging, electron paramagnetic resonance (EPR) and electrochemical measurements indicated that 1O2 and Fe(V)O were the main active species for SMX degradation in ß-CDPs/Fe-g-C3N4+PMS system. Moreover, ß-CDPs accelerated electron transfer between catalyst and PMS and promoted the generation of reactive oxygen species (ROS) during PMS activation. The loading of ß-CDPs increased the yields of Fe(V)O and 1O2 in the system and limited the leaching of Fe3+. In addition, the possible degradation pathways of SMX were described based on the intermediates detected by liquid chromatography-mass spectrometry (LC-MS), and the toxicity of the intermediates was also evaluated. This work investigate the role of ß-CDPs in PMS activation for the first time and develop a promising material with potential for water treatment.

Application of SifaInDel 45plex System in the Han and Mongolian Populations.

OBJECTIVES: To investigate the genetic polymorphism of InDel loci in SifalnDel 45plex system in the Han population in Jiangsu Province and the Mongolian population in Inner Mongolia, and to evaluate the effectiveness of the system in forensic medicine. METHODS: SifaInDel 45plex system was used for genotyping in blood samples of 398 unrelated individuals from the above two populations, and allele frequencies and population genetic parameters of the two populations were calculated respectively. Eight intercontinental populations in the gnomAD database were used as reference populations. The genetic distances between the two studied populations and eight reference populations were calculated based on the allele frequencies of 27 autosomal-InDels (A-InDels). The phylogenetic trees and multidimensional scaling (MDS) analysis diagrams were constructed accordingly. RESULTS: Among two studied populations, the 27 A-InDels and 16 X-InDels showed no linkage disequilibrium between each other and the allele frequency distributions were in Hardy-Weinberg equilibrium. The CDP of the 27 A-InDels in two studied populations were all higher than 0.999 999 999 9, and the CPEtrio were all less than 0.999 9. The CDP of the 16 X-InDels in Han in Jiangsu and Mongolian in Inner Mongolia female and male samples were 0.999 997 962, 0.999 998 389, and 0.999 818 940, 0.999 856 063, respectively. The CMECtrio were all less than 0.999 9. The results of population genetics showed that the Jiangsu Han nationality, Inner Mongolia Mongolian nationality and East Asian population clustered into one branch, showing closer genetic relationship. The other 7 intercontinental populations clustered into another group. And the above 3 populations displayed distant genetic relationships with the other 7 intercontinental populations. CONCLUSIONS: The InDels in the SifaInDel 45plex system have good genetic polymorphism in the two studied populations, which can be used for forensic individual identification or as an effective complement for paternity identification, and to distinguish different intercontinental populations.

Surgical management strategy of slide tracheoplasty for infants with congenital tracheal stenosis.

OBJECTIVE: The study objective was to evaluate the outcomes of slide tracheoplasty in infancy and identify predictors of adverse outcomes. METHODS: We retrospectively reviewed the clinical data of infants aged less than 1 year with congenital tracheal stenosis who underwent slide tracheoplasty at a single center from April 2010 to September 2020. RESULTS: Of 120 infants, 71.7% (86/120) had a pulmonary artery sling and 37.5% (45/120) had simultaneous intracardiac repairs. Additionally, 52.5% (63/120) of the patients had anomalous tracheobronchial arborization, and 17.5% (21/120) had diffuse tracheal stenosis. Six airway reoperations (5%) and 6 deaths (5%) occurred, and the mortality decreased annually. Multivariate analysis revealed that a low body weight, cardiovascular anomalies, and normal tracheobronchial arborization predicted a longer intubation duration. Univariate analysis revealed that a low body weight, preoperative invasive ventilation, a long cardiopulmonary bypass time, and granulation tissue were associated with death. After surgery, 26 patients had dysphagia, 24 of whom resumed oral feeding during follow-up. Ninety-two patients underwent chest computed tomography reexamination, and the trachea diameter had increased significantly from 2.32 ± 0.72 mm to 5.46 ± 1.24 mm. Nineteen and 29 patients underwent spirometry before and after surgery, respectively, and showed improvements in ventilation function, with the ratio of time to peak tidal expiratory flow to total expiratory time and ratio of volume to peak tidal expiratory flow to total expiratory volume values significantly improved from 19.80% (interquartile range, 16.90-23.80) and 23.10% (interquartile range, 21.10-25.90) to 26.80% (interquartile range, 21.20-34.40) and 30.20% (interquartile range, 25.00-34.50), respectively (P < .05). CONCLUSIONS: A tailored individual management strategy of slide tracheoplasty in infancy facilitates favorable clinical outcomes. Close postoperative follow-up and long-term functional evaluations including clinical symptoms and pulmonary function are still needed.

[Determination of three urinary catecholamines and serotonin by on-line packed-fiber solid-phase extraction].

Biogenic monoamines, including catecholamines (CAs) and serotonin (5-HT), play critical roles in the central nervous system. They have recently been proven to be primarily useful as biomarkers for the diagnosis of CA-producing tumors. The highly polar properties of biogenic monoamines result in poor retention on conventional materials, making it challenging to simultaneously measure more biogenic monoamines from complex matrices. Moreover, the classical method of off-line pretreatment is relatively complex, labor-intensive, and incurs errors in repeatability among different operators. Therefore, the development of an on-line sample pretreatment method combined with the use of specific nanofiber adsorbents has been explored. An on-line procedure could avoid unnecessary and time-consuming steps, and enable full automation of the experimental process. In this study, an on-line packed-fiber solid-phase extraction (PFSPE) and determination method for urinary CAs (dopamine (DA), norepinephrine (NE), epinephrine (E)) and 5-HT was developed, using composite nanofibers of polycrown ether-polystyrene (PCE-PS). PCE-PS composite nanofibers prepared by electrospinning were used as adsorbents in the PFSPE column, which was connected to the on-line HPLC system. The PFSPE-HPLC equipment contained a dual ternary pump and a switching valve to enable enrichment, purification, and analysis directly in the system. The left pump was connected with the PFSPE column for sample enrichment and purification, while the right pump was attached to the analysis column for sample separation and testing. The switching valve was controlled such that after enrichment, the samples could be eluted to the analysis column for separation and detection. The current work expands on our previous research by analyzing more target substances, and developing an on-line sample pretreatment method to simultaneously analyze four biogenic monoamines. Gradient separation aided in the satisfactory separation of the biogenic monoamines within a short retention time. The running time was set at 16 min to enable thorough enrichment, elution, and analysis. The influence of the complexing reagent (diphenylborinic acid 2-aminoethyl ester, 2 mg/mL) was also investigated with this on-line PFSPE-HPLC system. The results showed that the intensity of most analytes was significantly higher when 50 µL of the complexing reagent was added. The influence of a buffer on the extraction of the biogenic monoamines was also tested. The optimum extraction condition for the target analytes was achieved when artificial urine (AU) samples were diluted in a volume ratio of 1∶1 by phosphate- buffered saline solution (PBS, pH 7.8). Under the optimum experimental conditions, the on-line PFSPE-HPLC procedure showed good linearity (in the range of 1 ng/mL to 200 ng/mL) with correlation coefficients above 0.996 for the quantitative detection of urinary CAs (DA, NE, E) and 5-HT. For the CAs, the limit of detection (LOD) was 1 ng/mL (S/N=3), while the limit of quantitation (LOQ) was 2.5 ng/mL (S/N=10). For 5-HT, the LOD was 2.5 ng/mL (S/N=3) and the LOQ was 5 ng/mL (S/N=10). Moreover, high recovery rates and good reproducibility were obtained. The recoveries of AU and real urine spiked with CAs and 5-HT were in the range of 83.5%-117.7%, and the intra-day precision was lower than 10%. Additionally, no significant changes in the nanofibers were observed after repeated extraction, which reflected the good stability and reusability of the nanofibers. The nanofibers could be reused for more than 95 times. The on-line PFSPE-HPLC system was successfully applied for the determination of urinary CAs and 5-HT with good precision and high sensitivity. This high level of integration and automation was significantly advantageous in terms of its repeatability, as well as reduction in the time and effort required. The proposed on-line pretreatment and determination method can provide strong technical support for the detection and diagnosis of, as well as research on related diseases in clinical practice.

An Edaravone-Guided Design of a Rhodamine-Based Turn-on Fluorescent Probe for Detecting Hydroxyl Radicals in Living Systems.

The hydroxyl radical (·OH), one of the reactive oxygen species (ROS) in biosystems, is found to be involved in many physiological and pathological processes. However, specifically detecting endogenous ·OH remains an outstanding challenge owing to the high reactivity and short lifetime of this radical. Herein, inspired by the scavenging mechanism of a neuroprotective drug edaravone toward ·OH, we developed a new ·OH-specific fluorescent probe RH-EDA. RH-EDA is a hybrid of rhodamine and edaravone and exploits a ·OH-specific 3-methyl-pyrazolone moiety to control its fluorescence behavior. RH-EDA itself is almost nonfluorescent in physiological conditions, which was attributed to the formation of a twisted intramolecular charge transfer (TICT) state upon photoexcitation and the acylation of its rhodamine nitrogen at the 3' position. However, upon a treatment with ·OH, its edaravone subunit was converted to the corresponding 2-oxo-3-(phenylhydrazono)-butanoic acid (OPB) derivative (to afford RH-OPB), thus leading to a significant fluorescence increase (ca. 195-fold). RH-EDA shows a high sensitivity and selectivity to ·OH without interference from other ROS. RH-EDA has been utilized for imaging endogenous ·OH production in living cells and zebrafishes under different stimuli. Moreover, RH-EDA allows a high-contrast discrimination of cancer cells from normal ones by monitoring their different ·OH levels upon stimulation with ß-Lapachone (ß-Lap), an effective ROS-generating anticancer therapeutic agent. The present study provides a promising methodology for the construction of probes through a drug-guided approach.