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This review presents advances in the implementation of high - throughput se quencing and its application to the knowledge of medicinal plants. We conducted a bibliographic search of papers published in PubMed, Science Direct, Google Scholar, Scopus, and Web of Science databases and analyzed the obtained data using VOSviewer (versi on 1.6.19). Given that medicinal plants are a source of specialized metabolites with immense therapeutic values and important pharmacological properties, plant researchers around the world have turned their attention toward them and have begun to examine t hem widely. Recent advances in sequencing technologies have reduced cost and time demands and accelerated medicinal plant research. Such research leverages full genome sequencing, as well as RNA (ribonucleic acid) sequencing and the analysis of the transcr iptome, to identify molecular markers of species and functional genes that control key biological traits, as well as to understand the biosynthetic pathways of bioactive metabolites and regulatory mechanisms of environmental responses. As such, the omics ( e.g., transcriptomics, metabolomics, proteomics, and genomics, among others) have been widely applied within the study of medicinal plants, although their usage in Colombia is still few and, in some areas, scarce. (185)
El extracto de cloroformo (CE) y las fracciones obtenidas de las raíces de Aldama arenaria se evaluaron para determinar su actividad antiproliferativa in vitro contra 10 líneas ce lulares tumorales humanas [leucemia (K - 562), mama (MCF - 7), ovario que expresa un fenotipo resistente a múltiples fármacos (NCI/ADR - RES), melanoma (UACC - 62), pulmón (NCI - H460), próstata (PC - 3), colon (HT29), ovario (OVCAR - 3), glioma (U251) y riñón (786 - 0)]. CE presentó actividad antiproliferativa débil a moderada (log GI 50 medio 1.07), mientras que las fracciones 3 y 4, enriquecidas con diterpenos de tipo pimarane [ent - pimara - 8 (14), ácido 15 - dien - 19 - oico y ent - 8(14),15 - pimaradien - 3 ß - ol], presentaron activid ad moderada a potente para la mayoría de las líneas celulares, con un log GI 50 medio de 0.62 y 0.59, respectivamente. Los resultados mostraron una acción antiproliferativa in vitro prometedora de las muestras obtenidas de A. arenaria , con los mejores resul tados para NCI/ADR - RES, HT29 y OVCAR - 3, y valores de TGI que van desde 5.95 a 28.71 µg.mL - 1, demostrando que los compuestos de esta clase pueden ser prototipos potenciales para el descubrimiento de nuevos agentes terapéuticos
Subject(s)
Plants, Medicinal , High-Throughput Nucleotide Sequencing , Multiomics , Medicine, Traditional , Colombia摘要
La prueba prenatal no invasiva es un método de cribado de aneuploidías fetales y de resultar con riesgo alto debe ser confirmado a través de prueba genética diagnóstica. Es la prueba de detección más sensible y específica para las aneuploidías fetales comunes y minimiza la realización de técnicas invasivas, solo para las gestantes con riesgo elevado. Se debe realizar asesoramiento genético pre- y poscribado. Este estudio tiene como objetivo describir los fundamentos básicos de la prueba prenatal no invasiva mediante el análisis del ácido desoxirribonucleíco libre circulante en plasma materno para cribado de aneuploidías, y de los métodos primordiales y avances en biología molecular incluyendo las tecnologías de secuenciación de nueva generación, que lo han facilitado, considerando sus beneficios y limitaciones al aplicarla en la práctica clínica, en este campo que cambia con tanta rapidez(AU)
The non-invasive prenatal test is a screening method for fetal aneuploidies and if the result is at high risk, it must be confirmed through diagnostic genetic test. It is the most sensitive and specific detection test for common fetal aneuploidies and minimizes the use of invasive techniques, only for pregnant women at high risk. Genetic counseling should be performed before and after screening. This study aims to describe the basic fundamentals of non-invasive prenatal testing by analyzing free circulating deoxyribonucleic acid in maternal plasma for aneuploidy screening, and the primary methods and advances in molecular biology, including next-generation sequencing technologies, which have facilitated it, considering its benefits and limitations when applying it in clinical practice, in this rapidly changing field(AU)
Subject(s)
Humans , Female , Pregnancy , Plasma , DNA , Mass Screening , Prevalence , Risk Factors摘要
Glioblastomas are known for their poor clinical prognosis, with recurrent tumors often exhibiting greater invasiveness and faster growth rates compared to primary tumors. To understand the intratumoral changes driving this phenomenon, we employed single-cell sequencing to analyze the differences between two pairs of primary and recurrent glioblastomas. Our findings revealed an upregulation of ferroptosis in endothelial cells within recurrent tumors, identified by the significant overexpression of the NOX4 gene. Further analysis indicated that knocking down NOX4 in endothelial cells reduced the activity of the ferroptosis pathway. Utilizing conditioned media from endothelial cells with lower ferroptosis activity, we observed a decrease in the growth rate of glioblastoma cells. These results highlighted the complex role of ferroptosis within tumors and suggested that targeting ferroptosis in the treatment of glioblastomas requires careful consideration of its effects on endothelial cells, as it may otherwise produce counterproductive outcomes.
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ObjectiveTo explore the clinical features and causative genes of short stature children with unknown etiology， providing evidence for precise clinical diagnosis and treatment. MethodsThe study recruited children with suspected but undiagnosed short stature from the pediatric endocrinology department in our hospital between January 2018 and August 2022. A retrospective analysis was performed on the clinical manifestations， laboratory test and whole exome sequencing （WES） results. Causative genes were classified and analyzed according to different pathogenic mechanisms. ResultsA total of 48 children （30 boys and 18 girls） were enrolled， aged 7.73 ± 3.97 years， with a height standard deviation score （ HtSDS） of -3.63 ± 1.67. Of the patients， 33 （68.8%） suffered from facial anomalies， 31 （64.6%） from skeletal abnormalities， 26 ［54.2%， 61.5% of whom born small for gestational age （SGA）］ from perinatal abnormalities， 24 ［50.0%， 87.5% of whom with growth hormone （GH） peak concentration below normal］ from endocrine disorders and 21（43.8%） had a family history of short stature. Laboratory tests showed that GH peak concentration following stimulation test was （9.72 ± 7.25） ng/mL， IGF-1 standard deviation score was -0.82 ± 1.42， the difference between bone age and chronological age was -0.93 ± 1.39 years. Of the 25 cases with mutant genes found by WES， 14 （56.0%） had pathogenic mutation， 6 （24.0%） likely pathogenic mutation， and 5 （20.0%） mutation of uncertain significance. Pathogenic and likely pathogenic variants were identified in 14 genes， including 10 affecting intracellular signaling pathways （PTPN11， RAF1， RIT1， ARID1B， ANKRD11， CSNK2A1， SRCAP， CUL7， SMAD4 and FAM111A） and 4 affecting extracellular matrix （ECM） components or functions （ACAN， FBN1， COL10A1 and COMP）. ConclusionsA rare monogenic disease should be considered as the possible etiology for children with severe short stature accompanied by facial anomalies， disproportionate body types， skeletal abnormalities， SGA， GH peak concentration below normal and a family history of short stature. WES played an important role in identifying the monogenic causes of short stature. This study indicated that affecting growth plate cartilage formation through intracellular signaling pathways and ECM components or functions was the main mechanism of causative genes leading to severe short stature in children. Further research may help discover and study new pathogenic variants and gene functions.
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Objective To explore the clinical characteristics and treatment scheme of patients with spinal infection caused by Prevotella intermedia(P.intermedia).Methods Clinical diagnosis and treatment processes of a patient with spinal infection caused by P.intermedia admitted to the spinal surgery department of a hospital were summa-rized,and relevant literature was retrieved from database for reviewing.Results The patient,a 50 year old male,was admitted to the hospital due to"lumbago pain complicated with pain in double lower extremities for 2 months".The lesion tissue was taken for metagenomic next-generation sequencing(mNGS)detection,which detected P.in-termedia,and the patient was diagnosed with P.intermedia spondylitis.After treatments with open lesion clea-rance,tube rinsing+autologous bone transplantation fusion internal fixation,intravenous drip of ceftriaxone sodium and metronidazole,as well as metronidazole rinsing,infection was under control.A total of 16 available papers were retrieved,together with this case,a total of 17 patients were included,with 7 males and 10 females.The main risk factors were diabetes and history of corticosteroid use(35.3％).The most common invasion sites were lumbar ver-tebra(n=12)and thoracic vertebra(n=6).13 cases were positive for pathogen culture,3 cases were positive for molecular detection,and 1 case was positive for staining microscopy.17 patients received anti-anaerobic bacteria treatment,with 14 cases receiving combined surgical treatment.One case died,with a mortality of 5.9％;5 cases had partial neurological impairment,with a disability rate of 29.4％.The survival rate of patients who received treatment of anti-anaerobic bacteria combined with surgery was 92.8％,3 patients only with anti-anaerobic bacteria treatment but without surgery were all cured.Conclusion P.intermedia is an opportunistic pathogeanic bacteria which often causes infection in immunocomprised individuals and is prone to be misdiagnosed.It is recommended to perform mNGS detection to identify the pathogen as early as possible and seize the opportunity for treatment to reduce mortality.
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Objective To explore the application of metagenomic next-generation sequencing(mNGS)technology in the investigation of healthcare-associated infection(HAI)outbreaks of carbapenem-resistant Acinetobacter bau-mannii(CRAB).Methods Pathogenic detection by mNGS and conventional pathogen culture were performed on 5 patients in the intensive care unit(ICU)of a hospital from June 8 to 22,2023 from whom CRAB were detected.Microbial sampling was carried out in potentially contaminated environment.Bacterial culture,identification,and antimicrobial susceptibility testing were conducted.Comprehensive control measures were taken,and the effect was evaluated.Results The time required for reporting results by mNGS was shorter than the culture time([3.92± 1.05]days vs[6.24±0.25]days,P＜0.001).CRAB was isolated from the specimens of 5 patients.mNGS de-tected OXA-23 resistance genes from all patients.After comprehensive assessment by experts,4 patients were HAI and 1 patient was due to specimen contamination.According to the definition from Guidelines for HAI outbreak control,this event was considered an outbreak of HAI.The monitoring results of environmental hygiene showed that the detection rate of CRAB in the environment during the outbreak was 51.30％(59/115),mainly from the hands of health care workers and the surface of ventilators.After implementing multidisciplinary infection control measures,clinicians'hand hygiene compliance rate and implementation rate of ventilator disinfection increased from 40.83％(49/120)and 33.33％(16/48)to 82.61％(95/115)and 83.33％(30/36),respectively.The prognosis of patients was good,and no new case emerged during subsequent monitoring.The outbreak of HAI in this hospital has been effectively controlled.Conclusion mNGS is characterized by high precision,less time consumption,and high accuracy,and can be applied to the prevention and control of HAI outbreak and the study of antimicrobial-re-sistant genomes.It is of great significance for the anti-infection treatment of patients with multidrug-resistant orga-nism infection as well as the formulation of HAI prevention and control measures.Continuous improving disinfec-tion effectiveness and hand hygiene compliance is important for preventing and controlling CRAB infection.
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Objective To explore the diagnosis and clinical characteristics of atypical severe pneumonia caused by Chlamydia abortus(C.abortus).Methods Clinical data of 4 patients diagnosed with atypical severe pneumonia caused by C.abortus in a hospital from January 2021 to November 2022 were collected.Clinical characteristics,dia-gnosis and treatment,and precautions of the disease were comprehensively analyzed.Results All 4 patients were male,aged 63-73 years old,with acute onset,high fever,cough and expectoration.Three patients had a history of contact with poultry,one patient had a history of contact with abortion goat.The interval between the emerging of clinical symptoms and the onset of acute respiratory failure in 4 patients was 1-6 days,and the oxygenation index(PaO2/FiO2)at admission was less than 200 mmHg,which gradually decreased with the progression of the disease,active support with a ventilator was necessary.Two patients had an increase in white blood cell count,4 had an in-crease in neutrophil percentage,3 had a mild decrease in platelet count.Among 4 patients,2,2,3 and 4 patients showed elevated levels of aspartate aminotransferase,alanine aminotransferase,creatine kinase,and serum creati-nine respectively,2 patients had mild hyponatremia,4 patients showed significant increase in C-reactive protein,procalcitonin,and interleukin-6 levels.Four patients'chest CT findings showed main involvement of single or mul-tiple lung lobes,with exudation and consolidation,and later involvement of multiple lobes of lung.The metageno-mic next-generation sequencing of bronchoalveolar lavage fluid detected the DNA sequence of C.abortus.Based on the clinical manifestations,contact history,chest CT,and metagenomic next-generation sequencing results of 4 pa-tients,the diagnosis was C.abortus.atypical severe pneumonia.After timely adjustment of the treatment of anti-in-fection regimen based on doxycycline,the patients'condition improved and were discharged.Conclusion C.abor-tus may also cause human pneumonia,which can lead to serious clinical outcome after infection.Patient had a histo-ry of animal contact should be alert to such diseases.Metagenomic next-generation sequencing can detect C.abortus.
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Objective:To study the changes of intestinal flora before and after dietary intervention in pregnant women with gestational diabetes mellitus (GDM) and explore its correlation with the results of oral glucose tolerance test (OGTT), using third generation high-throughput sequencing of full-length 16S rDNA.Methods:Thirty pregnant women who received regular antenatal care in the First Affiliated Hospital of Xinjiang Medical University between July 2022 and March 2023 were selected. Demographic data and laboratory examination results of these pregnant women were collected. The pregnant women with GDM (the GDM group) were given individualized dietary intervention, while the control group were given routine dietary guidance, for a total of 2 weeks. The stool was collected before and after the intervention in both groups, and a total of 60 stool samples were collected. Through the PacBio Sequencing platform, the third generation sequencing of full-length 16S rDNA was utilized to investigate the intestinal microbiome diversity. The composition, abundance and diversity of intestinal flora were compared between groups, both before and after the intervention.Results:There were 74 species showing significant differences in abundance between the two groups of pregnant women, 54 of which were enriched in the GDM group. The correlation analysis of blood glucose levels tested by OGTT as an environmental factor showed that Lachnospirales ( P=0.002) and unclassified Bacteria ( P=0.035) were positively correlated with OGTT-0h blood glucose ( P<0.05), while Christensenellales ( P=0.018)and Oscillospirales ( P=0.045) negatively. Lachnospirales ( P=0.027)and unclassified Bacteria ( P=0.028) were positively correlated with OGTT-1 h blood glucose ( P<0.05), while Oscillospirales ( P=0.025) negatively. There was a positive correlation between Lachnospirales ( P=0.027) and OGTT-2h blood glucose ( P<0.05). Conclusions:After the individualized diet intervention, the dominant bacteria of the intestinal flora changed and the fasting blood glucose of declined in pregnant women with GDM. It's a reasonable presumption that individualized diet intervention can contribute to the regulation of the disordered intestinal flora in pregnant women with GDM, which implies a role of dietary intervention in the treatment of GDM.
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The ICR(Institute of Cancer Research)mouse infection model was constructed to study the pathogenicity of Sal-monella Telelkebir serotype,and the pathogenic identification of mouse isolates was carried out.Observe the bacterial excretion cycle,evaluate the pathogenicity of Salmonella serotype to mice,and calculate the LD50 by the changes in clinical characteris-tics,histopathology and tissue bacterial load of infected mice;by flight mass spectrometry,biochemical identification,serotype identification,molecular typing and other experiments,compared with human isolates;virulence gene analysis was carried out by PCR experiment and whole genome sequencing.The LD50 of Salmonella Telelkebir is 2.67 × 108 CFU/mL;curling and fluffing may occur 0.5 h after infection;autopsy of dead mice showed that the small intestine was severely congested,with more bubbles and fluid accumulation,cecal necrosis,liver apical degeneration and necrosis,necrotic foci on the surface of the kidney and spleen atrophy;the bacterial load of spleen,kidney,lung,liver and jejunum in mice reached its peak at 3 days after infection,while that of heart at 6 days;the bacterial excretion time of the high-dose group exceeded 100 days;The level of CD3 in tissues increased with increasing dose,with inflammatory cell infiltration,myocardial capillary dilation and hyperemia,large area of vacuoles,degeneration and necrosis of hepatocytes,obvious enlargement of splenic sinus,blurred zoning,thickening of glomerular basement membrane,partial exfoliation of ciliated epithelium,atrophy and exfoliation of jejunal villi;PCR and whole genome sequencing revealed Salmonella-related virulence genes such as cdtB,plt A and pltB.This study was the first to successfully establish the ICR mouse model of Salmonella Telelkebir,demonstrating that this serotype of Salmonella has some pathogenicity.
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The aim of this study was to investigate the virulence determinants and genetic diversity of foodborne Yersinia enterocolitica from Wenzhou.A total of 71 strains of Yersinia enterocolitica were isolated from food and foodborne diarrhea ca-ses in Wenzhou,and their biotypes,serotypes,and drug resistance were analyzed.On the basis of whole genome sequencing,we assessed virulence gene profiles,and performed multilocus sequence typing(MLST)and core gene multilocus sequence typ-ing(cgMLST).A total of 94.4％(67/71)of isolates belonged to biotype 1A,and the highest proportion had serotype lA/O∶5(29.6％,21/71).The sensitivity rates of the isolates to 14 antibiotics exceeded 95.8％.A total of 16 categories and 126 viru-lence genes were identified,with two strains carrying the pYV plasmid and chromosome-related virulence genes.ST3(31.6％,12/38)was the most widespread MLST type,and cgMLST analysis revealed no dense clusters of genotypes except for strains sharing the same ST.In conclusion,pathogenic strains were identified from foodborne Yersinia enterocolitica in Wenzhou and were found to exhibit high genetic polymorphism.Enhanced regulatory supervision is essential to prevent the outbreak of food-borne diseases caused by Yersinia enterocolitica.
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We aimed to identify the infectious source of a case of melioidosis,to provide evidence for the prevention and control of melioidosis in Shanxi Province,China.The patient developed repeated fever,fatigue,diarrhea,and other symptoms after being caught in the rain while traveling in Hainan Province.The blood culture was positive,and the bacterial strain was i-dentified as Burkholderia thayensis and sent to the provincial Center for Disease Control and Prevention for further evaluation.MALDI-TOF MS and biochemical identification were used to identify the strain,whole genome sequencing was performed after nucleic acid extraction,MLST type and drug-resistance genes were analyzed,and a phylogenetic tree was constructed.The iso-lated strain was identified as Burkholderia pseudomallei by MALDI-TOF MS and biochemistry,and the MLST type was 366.The whole gene sequencing analysis indicated a close evolutionary relationship with the three isolates in Hainan Province,with high homology.This case of melioidosis was indeed imported from Hainan Province,according to molecular tracing analysis and epidemiological investigation,thus suggesting that medical institutions and disease control departments should strengthen understanding of melioidosis,and improve the diagnosis and treatment ability.
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This study was aimed at performing epidemiologic investigation of the first psittacosis death case in Hangzhou City,to provide a reference for the investigation and disposal of psittacosis cases,as well as prevention and control.Epidemio-logic data were collected through field epidemiologic investigation,and close contacts and environmental samples were collected for pathogenicity testing.The first symptom in the patient was cough,which did not raise concerns at the time.Several days later,the patient developed abdominal distension and black stools,and visited two medical institutions for treatment and hospi-talization.The patient's sputum and peripheral blood were tested for Chlamydia psittaci infection by metagenomic analysis via next-generation sequencing.Samples collected from the patient's family members,close contacts,and home environment test-ed negative with real-time quantitative polymerase chain reaction.The patient later died of gastrointestinal bleeding.This article is the first report of a case of psittacosis contracted from exposure to a sick parrot in Hangzhou City,in a patient who died be-cause of an underlying disease.Operational training should be provided for medical personnel,and early diagnosis with mNGS and treatment of patients with underlying diseases should be performed as early as possible to avoid fatality.In addition,health education should be carried out to raise public awareness of the disease.
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【Objective】 To study the molecular mechanism of 95 samples of serological ABO subtypes. 【Methods】 A total of 95 samples with discrepancy between forward and reverse blood grouping were subjected to serological confirmation, and genotyped by polymerase chain reaction with sequence-specific primers (PCR-SSP). For those subtype alleles could not be detected by PCR-SSP, ABO gene exon 1-7 sequencing and gene single strand sequencing were performed successively to determine the mutation site and the gene location. 【Results】 A total of 34 ABO alleles were detected in 95 samples. Five common ABO alleles (ABO*A1.01, ABO*A1.02, ABO*B.01, ABO*O.01.01 and ABO*O.01.02) and 29 rare ABO alleles were identified, including 16 named alleles by ISBT (ABO*A2.01, ABO*A2.05, ABO*A2.13, ABO*A3.07, ABO*AW.37, ABO*AEL.05, ABO*B3.01, ABO*B3.05, ABO*BW.03, ABO*BW.07, ABO*BW.27, ABO*BEL.03, ABO*cisAB.01, ABO*cisAB.05, ABO*BA.02, ABO*BA.04) and 5 named alleles by dbRBC(A223, B309, Bw37, Bel09, Bw40)and eight unnamed alleles [ABO*B.01+ 978C>A, ABO*A1.02+ 248A>T, ABO*B.01+ 125dupT, ABO*B.01+ (98+ 1G>A), ABO*A1.02/ABO*B.01+ 1A>G, ABO*A1.02/ABO*O.01.01+ 28G>T, ABO*A1.02/ABO*B.01+ 538C>T, ABO*A1.02/ABO*O.01.01+ 797insT] .The last four samples could not be verified by single strand because of insufficient samples. In 95 samples, 76 samples (21 named alleles of ISBT and dbRBC) were identified by PCR-SSP, and the remaining 19 samples were identified by exon 1-7 sequencing of ABO gene, of which 8 were identified as unnamed alleles, and the remaining 11 samples were not identified as subtype alleles. 【Conclusion】 The molecular genetic mechanism of 95 serological ABO subtypes was revealed, and 8 rare novel alleles were identified. The detection of ambiguous blood groups is influenced by factors such as patient pathology and physiology, therefore the combination of serological testing and genetic testing is suggested for the identification of ABO subtype.
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【Objective】 To investigate and analyze the serological and molecular biological characteristics of B(A) subgroup in a tertiary hospital in Jiaozhou, Qingdao. 【Methods】 From November 2019 to February 2023, the samples of 12 patients were suspected to be AB subgroup by microcolumn glass bead method and saline test tube method. The exons 6 and 7 of ABO gene were further amplified, sequenced and analyzed to determine the ABO allele type. 【Results】 A total of 9 cases of B(A) subgroup were detected in 26 065 patients in Jiaozhou, with a detection rate of 0.345 ‰ ( 9/26 065 ). Among the 9 cases of B(A) subgroup, 8 cases of serological reaction showed AweakB, and the gene detection was heterozygous for BA.04 gene and O gene.One case of serological reaction showed ABweak, and the gene detection was heterozygous for BA.04 gene and A gene. 【Conclusion】 Blood group serological combined with gene detection can accurately identify ABO blood group. B(A) subgroup alleles can exist in individuals with serological reaction of ABweak.
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Objective To explore the effects of different types of drinking water on the growth and fecal flora of mice.Methods Specific pathogen-free NIH mice were randomly divided into five groups,32 mice each group,with half males and half females in each group.The group were given either purified water(control group),acidified water,alkalized water,weakly acidic water or solid water.Diet and body weight were monitored continuously for 20 days.After the experiment,animal fecal samples were collected,and the V3-V4 region was amplified with bacterial 16S rDNA universal primers.An Illumina Miseq high-throughput sequencing platform was used for high-throughput sequencing,and microbial community,α diversity and β diversity were analyzed by bioinformatics method.Results The body weight of female mice given different pH values of weakly acidic water was higher,while the weight of the other groups was lower,than that of the control group(P＞0.05).The body weight of male mice in the acidified water group was higher,while that of other groups was lower,than that of control group,but there was no statistical difference between the groups(P＞0.05).The body weights of male and female mice in the solid water group were lower than those in the control group(P＜0.05).The food and water intake of the female animals in the alkaline water group and the water intake of female animals in the solid water group were lower than those of the other groups.OTU clustering analysis showed that the data volume of the sequencing was reasonable,and the fecal flora species of NIH mice were divided into five phyla,among which Bacteroides and Firmicutes were dominant.Unclassified Pseudopurpuromonas,Lactobacillus and Alistipes were the main genera.There were differences in fecal flora abundance and diversity among the mice given the five drinking water types.α analysis showed that the acidified water group had the highest flora abundance and diversity,while the solid water group had the lowest flora diversity.β analysis showed that the fecal flora composition in the solid water group was the closest to that of the control group,followed by the alkalized water group,acidified water group and weakly acidic water group.Conclusions Through an exploration of the effects of consuming different forms of water,this study revealed that solid water consumption had the greatest effects on body weight,feed intake,water consumption,and fecal flora of mice.The abundance and diversity of fecal flora in mice were affected by different pH values of drinking water,especially acidified water.
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Objective To investigate the effect of vitamin D(VD)on intestinal flora in spontaneously diabetic rats.Methods Zucker diabetic fatty rats(ZDF rats)were randomised to control(Con)group,VD control(VD)group,model(T2DM)group,and VD intervention(VD+T2DM)group.Fasting blood glucose profiles and oral glucose tolerance levels were determined in rats of each group.16S rDNA sequencing was used to assess changes in rat intestinal flora.OTU analysis(Venn diagram),α diversity analysis(chao1,observed species,PD whole tree,and shannon and simpson),βdiversity analysis(principal coordinate analysis(PCoA)),flora structure,and colony species variability analysis(linear discriminant analysis and influence factor(LEfSe)analysis)were also performed.Results VD intervention significantly improved fasting blood glucose levels and insulin resistance in T2DM rats(P＜0.05).α diversity showed no significant differences in chao1,observed species,PD whole tree,and shannon and simpson indices between T2DM and VD+T2DM groups(P＞0.05).β diversity analysis showed that the VD+T2DM group had more species similarity to the Con group than the T2DM group.The dominant bacteria of rat intestinal flora in each group were significantly different.In comparison to the T2DM group,the VD+T2DM group showed a decrease in abundance of Bacteroidetes and increases in abundances of Firmicutes and Clostridium XIVa.Conclusions VD improves fasting glucose elevation and insulin resistance in T2DM rats.VD improves the structure of intestinal flora,decreases Bacteroidetes,and elevates Firmicutes and Clostridium XIVa abundances in T2DM rats.
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Objective To examine the differential expression of plasma circRNA in individuals with type 2 diabetes mellitus(T2DM)and to elucidate the potential role of circRNA in the pathogenesis and progression of T2DM.Methods A total of 33 newly diagnosed T2DM patients who were hospitalized in the Department of Endocrinology of Heze Municipal Hospital and 33 healthy subjects with normal glucose tolerance(NC)were enrolled in this study from March 2022 to March 2023.Three subjects with T2DM and 3 with NC were randomly selected from study population and underwent high-throughput sequencing to identify differentially expressed circRNA.Six circRNA with significant differences were selected,and validated in the remaining study population using Real-Time Quantitative Polymerase Chain Reaction(RT-qPCR).Bioinformatics analyses were conducted on the differentially expressed circRNA-associated genes,and their interaction with microRNA(miRNA)was predicted.Results Microarray analysis revealed 166 significantly differentially expressed circRNA(FC≥2,P<0.05)in the T2DM group compared with NC group.Among them,137 were up-regulated and 29 were down-regulated.RT-qPCR validation of six circRNA showed that the expression level of hsa_circ_0000705 was significantly higher,while the expression levels of hsa_circ_0005362 and hsa_circ_0042839 were significantly lower in T2DM group,consistent with the microarray results.However,hsa_circ_0117392,hsa_circ_0008311,and hsa_circ_0087641 showed no significant changes.Conclusion Differentially expressed circRNA are present in T2DM patients.Hsa_circ_0005362 was significantly down-regulated and may be involved in the development of T2DM through the regulation of related signaling pathways by targeting miR-128-1-5p.
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Objective:To explore the application value of Oxford nanopore technologies (ONT) in the diagnosis and treatment of periprosthetic joint infection (PJI).Methods:A prospective analysis was conducted on 32 patients with PJI admitted to the joint department of Xi'an Honghui Hospital from October 2021 to March 2023, who met the 2018 PJI diagnostic criteria of the American Skeletal Infection Society (MSIS), including 15 males and 17 females with an average age of 63.93±8.93 years. 32 revision patients who did not meet the 2018 MSIS PJI criteria during the same period were collected as controls (non PJI group), including 13 males and 19 females with an average age of 65.53±8.54 years. All patients underwent joint fluid puncture before or during surgery, and the specimens were tested by ONT, metagenomic next generation sequencing (mNGS), and general microbial culture. The receiver operating characteristic (ROC) curves were drawn for both groups, and the sensitivity, specificity, positive predictive value, negative predictive value, and Youden index of the three detection techniques were calculated and compared to evaluate the detection efficiency of different detection methods in PJI.Results:Among the 32 patients with PJI, 30 were positive for ONT, with a total of 30 pathogenic bacteria detected, and the detection time was 22.37±8.36 h. 31 were positive for mNGS, with a total of 33 bacterial species detected, and the detection time was 46.25±9.36 h. 17 were positive for microbial culture, with a total of 8 bacterial species detected, and the detection time was 96.23±15.62 h. Among the 32 patients with non PJI group, 1 was positive for ONT and 5 were positive for mNGS, with a total of 1 and 3 bacterial species detected, respectively. The results of microbial culture were all negative. The detection time and area under the curve (AUC) of ONT and mNGS were 22.37±8.36 h and 0.953[95% CI (0.901, 1.006)], 46.25±9.36 h and 0.906[95% CI (0.835, 0.977)], respectively, which were better than those of microbial culture 96.23±15.62 h and 0.766[95% CI (0.678, 0.853)], and the difference was statistically significant ( P<0.05). The sensitivity of ONT, mNGS, and microbial culture were 0.938, 0.969, and 0.531, respectively, and the specificity was 0.969, 0.844, and 1.000, respectively. The Jordan index was 0.906, 0.813, and 0.531, respectively. Conclusion:ONT testing has higher diagnostic efficacy than mNGS and microbial culture in the diagnosis of PJI, and also has advantages in detection time. It also suggests that some PJI are not caused by a single microbial infection.
摘要
Rhino-orbito-cerebral mucormycosis (ROCM) is an acute, rapidly progressive, and lethal opportunistic fungal disease. Due to the atypical clinical manifestations, the disease is easily misdiagnosed in the early stage. The patients with ROCM associated cerebrovascular complications generally have a high mortality rate. This article reports a 47-year-old female patient with diabetic ketoacidosis and COVID-19 admitted to Central Hospital Affiliated to Shandong First Medical University. The results of radiological examinations and cerebrospinal fluid metagenomic next-generation sequencing confirmed as Rhizopus oryzae associated ROCM. In spite of receiving amphotericin B colloidal dispersion and isavuconazole treatments, the patient died of ROCM complicated with severe cerebral infarctions and pulmonary infection. The purpose of this case report is to summarize the clinical characteristics of ROCM with cerebrovascular ischemic events and the rare condition of bilateral anterior circulation involvements, and introduce recent diagnostic and therapeutic approaches for this disease.
摘要
Objective:To investigate the clinical phenotype and genetic characteristics of developmental epileptic encephalopathy 18 (DEE18) caused by SZT2 gene variants. Methods:Clinical data of 2 children with SZT2 related DEE18 who visited the Department of Pediatric Neurology, Linyi People′s Hospital in March 2020 and July 2023 were collected. The whole exome sequencing (WES) and Sanger sequencing were applied to verify the child and their parents. SWISS-MODEL software was used to perform protein 3D modeling for the selected SZT2 gene variants. Results:Both of the 2 cases showed severe global developmental delay, epileptic seizures, autism, megacephaly, facial deformity, hypotonia, corpus callosum malformation, persistent cavum septum pellucidum, and slow background activity and focal discharge in video electroencephalography. Case 1 was easy to startle and thin in stature; case 2 had immune deficiency and clustered seizures. WES results showed that case 1 carried a compound heterozygous variant of c.5811G>A (p.W1937X) (paternal) and c.9269delG (p.S3090Ifs *94) (maternal), while case 2 carried a compound heterozygous variant of c.6302A>C(p.H2101P) (paternal) and c.7584dupA (p.E2529Rfs *20) (maternal), the parents of both patients with normal clinical phenotypes. The 4 mutations mentioned above were novel variations that had not yet been reported domestically or internationally. According to the American College of Medical Genetics and Genomics variant classification criteria and guidelines, the p.S3090Ifs *94 variant was interpreted as pathogenic; p.W1937X variant was interpreted as pathogenic; p.E2529Rfs *20 variant was interpreted as likely pathogenic; p.H2101P variant was interpreted as uncertain significance. 3D modeling showed that the variant of p.H2101P resulted in a significant change in the hydrogen bond around the 2 101st amino acid encoded, leading to a decrease in protein stability. The other 3 variants led to early truncation of peptide chain and obvious changes in protein structure. Conclusions:DEE18 caused by SZT2 gene mutation is mainly an autosome recessive genetic disease, and its clinical manifestations include global developmental delay, epileptic seizures, autism, craniofacial malformation, hypotonia, epileptic discharge, corpus callosum malformation, persistent cavum septum pellucidum, shock, small and thin stature, and immune deficiency. Four novel variants related to the SZT2 gene may be the genetic etiology of DEE18 patients in this study.