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The enzyme dihydropyrimidine dehydrogenase (DPD) is the primary catabolic pathway of fluoropyrimidines including 5 fluorouracil (5FU) and capecitabine. Cases of lethal toxicity have been reported in cancer patients with complete DPD deficiency receiving standard dose of 5FU or capecitabine. DPD is encoded by the pharmacogene DPYD in which more than 200 variants have been identified. Different approaches have been developed for screening DPD-deficiency, including DPYD genotyping and phenotyping. Plasma uracil ([U]) and dihydrouracil ([UH2]) concentrations are routinely used as surrogate markers for systemic DPD activity: [U] ≥ 16 ng/ml and < 150 ng/ml, and [U] ≥ 150 ng/mL indicate partial and complete DPD deficient phenotype, respectively, while values of 5 or 10 for [UH2]/([U] ratio are often cited. Four clinically relevant DPYD defective variants (DPYD*13, DPYD*2A, p.Asp949Val and haplotype B3), are targeted in genetic testing via PCR. In practice, pretreatment [U], alone or combined with these 4 recommended DPYD alleles guides individual dosage selection, though this approach has limitations. This is illustrated by two cases showing discrepancy between DPD deficient phenotype and normal standard genotype. In these two cases, DPYD exome sequencing with Next Generation Sequencing identified rare inactive variants, establishing concordance between phenotype and genotype. In patient 1, [U] levels of 21.1 and 25.5 ng/mL, indicated partial deficiency though the targeted genotype was normal and 5FU dose was adjusted based on the phenotype. In patient 2, [U] levels of 16.2 and 15.2 ng/mL were near the 16 ng/ml threshold. With a normal genotype, he as considered non-deficient as targeted genotype was normal and the standard dose was administered. These two cases underscore the need to pair DPD phenotyping with whole DPYD gene sequencing, due to the frequent discrepancies between these pharmacogenetic tools, the burden of rare variants and ethnic differences in variant frequencies. [ABSTRACT FROM AUTHOR]

Methane (CH4) processes and fluxes have been widely investigated in low-latitude tropical wetlands and high-latitude boreal peatlands. In the mid-latitude Mongolia Plateau, however, CH4 processes and fluxes have been less studied, particularly in riverine wetlands. In this study, in situ experiments were conducted in the riverine sandy wetlands of the Mongolia Plateau to gain a better understanding of CH4 emissions and their influencing mechanisms. Annual CH4 emissions were observed at 8.7 mg m–2 h–1 from the flowing water wetlands during November 2019 − October 2021, approximately 80% and 20% of which were emitted during the growing and non-growing seasons, respectively. In particular, CH4 emissions during the thawing period contributed < 5% to the annual total, contrary to the traditional idea that thawing plays an important role in annual CH4 emissions in boreal peatlands. CH4 emissions were significantly higher in the wetlands dominated by plant species than in that dominated by water body during the growing seasons; therefore, plant-mediated CH4 transport was explained as a favorable pathway for CH4 emissions from sandy soils to the atmosphere. Gene sequencing revealed differences in the phylogenies and taxonomies of methanogenic archaea and methanotrophs between the flowing and static water wetlands, suggesting that flowing water should bring oxygen and nutrients to microbial habitats and potentially affect the production, oxidation, and diffusion of CH4 in sandy wetlands. [ABSTRACT FROM AUTHOR]

Objective To investigate the concentration and genetic variations of SARS-CoV-2 in sewage in Guiyang city from February to December 2023. Methods A total of 1 034 sewage samples were analyzed for ORFlab and N gene concentrations using quantitative PCR, and full viral genome sequencing was performed using the Illumina sequencing platform. Results Among the 1 034 samples collected, 587 were tested positive for SARS-CoV-2. The median concentration of the ORFlah gene was 3.69 copies/ml, and the median concentration of the N gene was 7.57 copies/ml. There was a correlation between the concentration of SARS-CoV-2 in sewage and the number of reported cases from medical institutions. Full genome sequencing was conducted on five SARS-CoV-2 positive samples with CT values <32, achieving an average coverage ranging from 63.91% to 97.28%. All viral types identified were Omicron sub lineages, with a total of 110 mutation sites identified. The S protein exhibited 22 amino acid changes, including the critical mutation E484A. Conclusion The detection rate of SARS-CoV-2 in urban sewage in Guiyang city is relatively high, and the viral concentration correlates well with clinical cases. The application of whole genome sequencing technology allows for the detection of genetic variants in the viral material present in sewage and identifies multiple mutation sites. Monitoring viruses in urban sewage effectively captures the spatiotemporal trends of the disease, serving as an important complement to clinical testing and providing a reference for local governments to implement precise epidemic prevention policies. [ABSTRACT FROM AUTHOR]

Simple Summary: The Yunnong (YN) chicken is a local Chinese breed known for its egg-laying capabilities. This study aimed to identify genes associated with eggshell quality in YN chickens by comparing them with the commercially established Jingfen (JF) breed. The results showed that YN chicken eggs had superior eggshell thickness, strength, mammillary density, and effective layer thickness compared to JF eggs. RT-qPCR analysis identified five key genes—LSS, NSDHL, MSMO1, SQLE, and FDFT1—linked to eggshell quality. Pathway enrichment analysis revealed a complex interplay of metabolic, immune, and cellular processes potentially influencing eggshell traits in poultry. The eggshell quality traits in avian species, including Yunnong chickens (YN), are crucial for commercial and breeding purposes. The use of advanced biotechnologies, such as gene sequencing, has become increasingly common for identifying genetic markers and comparing specific traits in livestock. However, genetic markers related to eggshell traits in YN chickens remain unexplored. This study aimed to compare eggshell quality traits and identify genetic markers in YN chickens versus commercial Jingfen chickens (JF). A total of 400 chicks (200 per breed) were reared for 300 days under controlled conditions. At the end, 60 eggs from each breed were analyzed using scanning electron microscopy to examine eggshell cross-sections and mammillary layer structure. Tissue samples from kidneys and eggshell glands were collected from six hens per breed for RT-qPCR analysis to study gene expression profiles. Results showed that YN chickens had significantly higher eggshell strength, thickness, mammillary density, and effective layer thickness than JF chickens (p < 0.01). Despite this, YN chickens had a thinner mammillary layer. RT-qPCR analysis confirmed five candidate genes related to eggshells (LSS, NSDHL, MSMO1, SQLE, and FDFT1) that play an important role in this process. Pathway analysis revealed significant enrichment in several biological processes, including steroid biosynthesis, glycerolipid metabolism, purine metabolism, and thiamine metabolism. Based on these results, the YN chickens have strong eggshells with better thickness, mammillary density, and effective layer thickness. In addition, genomic and pathways analysis suggest a complex interaction of metabolic, immune, and cellular processes potentially influencing eggshell quality traits in poultry. [ABSTRACT FROM AUTHOR]

Actinomycetota are bacteria with biotechnological potential known to produce bioactive compounds in their natural ecosystems. Natural ecosystems include extreme environments such as soda lakes. Lake Natron is one of the soda lakes found in Arusha region, Tanzania. This study aimed to characterise and establish the phylogenetic diversity of the isolated Actinomycetota species that inhabit the lake waters. Results after 16S rRNA gene sequencing showed eleven genera of phylum Actinomycetota: Streptomyces, Micrococcus, Microbacterium, Gordonia, Micromonospora, Kocuria, Isoptericola, Dietzia, Arthrobacter, Nocardiopsis and Mycobacterium have been identified. Of these, Streptomyces radiopugnans, Streptomyces harenosi, Streptomyces althioticus, Micrococcus terreus and Kocuria sp. are reported for the first time in this soda lake, and this is the first time that Gordonia sp. has been reported from a soda lake in East Africa. Micromonospora sp. and Arthrobacter sp. are reported for the first time as occurring in a Tanzanian soda lake and three putative new species of Actinomycetota have been revealed. The Actinomycetota species described in this study sets a platform for future bioprospection of novel bioactive compounds from Lake Natron. [ABSTRACT FROM AUTHOR]

This article presents the first record of Rüppell's Bat (Vansonia rueppellii) within Saudi Arabia, captured within Prince Mohammed bin Salman Royal Reserve in northwestern Saudi Arabia. The morphological and acoustic description is provided. The species has been confirmed by biometrics and genetic sequencing, increasing the number of bat species currently found within Saudi Arabia to 32, from 23 genera and 10 families. [ABSTRACT FROM AUTHOR]

Hypophosphatasia (HPP) is a rare, inherited metabolic disease characterized by low tissue‐nonspecific alkaline phosphatase activity due to ALPL gene variants. We describe ALPL variants from the observational, prospective, multinational Global HPP Registry. Inclusion in the analysis required a diagnosis of HPP, low serum ALP activity, and ≥1 ALPL variant. Of 1176 patients enrolled as of September 2022, 814 met inclusion criteria in Europe (48.9%), North America (36.7%), Japan (10.2%), Australia (2.6%), and elsewhere (1.6%). Most patients (74.7%) had 1 ALPL variant; 25.3% had ≥2 variants. Nearly all patients (95.6%) had known disease‐causing variants; 4.4% had variants of uncertain significance. Disease‐causing variants were predominantly missense (770/1556 alleles). The most common variants were c.571G>A (102/1628 alleles), c.1250A>G (66/1628 alleles), and c.1559del (61/1628 alleles). Variant profiles were generally consistent, except in Japan, where a higher proportion of patients (68.7%) had ≥2 ALPL variants, likely because more had disease onset before age 6 months (53.0% vs. 10.1%–23.1% elsewhere). Frameshift mutations (61/164 alleles) and inframe deletions (7/164 alleles) were more common in Japan. Twenty‐three novel variants were discovered, each in a single geographic region, predominantly Europe. Analyses confirmed previously known ALPL variants, identified novel variants, and characterized geographic variation in frequency and type of ALPL variants in a large population. [ABSTRACT FROM AUTHOR]

Objective To investigate the serological characteristics and molecular mechanism of a case of Bel subtype. Methods Serological method was used to analyze the ABO blood group of the patient, absorption and elution test was used to detect the weak antigen on red blood cells, and molecular biology method was used to analyze the ABO gene sequence of the patient. Results The forward typing of the patient was O type, but the reverse typing was B type. The weak B antigen was confirmed by absorption and elution method. This mutation was not found in ISBT and related literature, and was discovered for the first time. The patient's blood type was Bel (29insertG)/O.01.01 based on the sequence of exon 3-7. Conclusion In the case of ABO blood group incompatibility, the combination of serological and molecular biological detection techniques can improve the accuracy and reliability of ABO blood group identification.

With rapid development and maturity of single-cell measurements, single-cell biology and pathology has become an emerging discipline to understand the disease. However, it is important to address concerns raised by clinicians as to how to apply single-cell measurements for clinical practice, translate the signals of single-cell systems biology into determination of clinical phenotype, and predict patient response to therapies. This paper proposes a new system coined as the clinical artificial intelligent single-cell (caiSC) with the dynamic generator of clinical single-cell informatics, artificial intelligent analyzers, molecular multimodal reference boxes, clinical inputs and outputs, and AI-based computerization. This system provides reliable and rapid information for impacting clinical diagnoses, monitoring, and prediction of the disease at the single-cell level. The caiSC represents an important step and milestone to translate the single-cell measurement into clinical application, assist clinicians’ decision-making, and improve the quality of medical services. There is increasing evidence to support the possibility of the caiSC proposal, since the corresponding biotechnologies associated with caiSCs are rapidly developed. Therefore, we call the special attention and efforts from various scientists and clinicians on the caiSCs and believe that the appearance of the caiSCs can shed light on the future of clinical molecular medicine.

Abstract Background Endophytic fungi (EF) reside within plants without causing harm and provide benefits such as enhancing nutrients and producing bioactive compounds, which improve the medicinal properties of host plants. Selecting plants with established medicinal properties for studying EF is important, as it allows a deeper understanding of their influence. Therefore, the study aimed to investigate the impact of EF after inoculating the medicinal plant Perilla frutescens, specifically focusing on their role in enhancing medicinal properties. Results In the current study, the impact of two EF i.e., Irpex lenis and Schizophyllum commune isolated from A. bracteosa was observed on plant Perilla frutescens leaves after inoculation. Plants were divided into four groups i.e., group A: the control group, group B: inoculated with I. lenis; group C: inoculated with S. commune and group D: inoculated with both the EF. Inoculation impact of I. lenis showed an increase in the concentration of chlorophyll a (5.32 mg/g), chlorophyll b (4.46 mg/g), total chlorophyll content (9.78 mg/g), protein (68.517 ± 0.77 mg/g), carbohydrates (137.886 ± 13.71 mg/g), and crude fiber (3.333 ± 0.37%). Furthermore, the plants inoculated with I. lenis showed the highest concentrations of P (14605 mg/kg), Mg (4964.320 mg/kg), Ca (27389.400 mg/kg), and Mn (86.883 mg/kg). The results of the phytochemical analysis also indicated an increased content of total flavonoids (2.347 mg/g), phenols (3.086 mg/g), tannins (3.902 mg/g), and alkaloids (1.037 mg/g) in the leaf extract of P. frutescens inoculated with I. lenis. Thus, overall the best results of inoculation were observed in Group B i.e. inoculated with I. lenis. GC-MS analysis of methanol leaf extract showed ten bioactive constituents, including 9-Octadecenoic acid (Z)-, methyl ester, and hexadecanoic acid, methyl ester as major constituents found in all the groups of P. frutescens leaves. The phenol (gallic acid) and flavonoids (rutin, kaempferol, and quercetin) were also observed to increase after inoculation by HPTLC analysis. The enhancement in the phytochemical content was co-related with improved anti-oxidant potential which was analyzed by DPPH (% Inhibition: 83.45 µg/ml) and FRAP (2.980 µM Fe (II) equivalent) assay as compared with the control group. Conclusion Inoculation with I. lenis significantly enhances the uptake of nutritional constituents, phytochemicals, and antioxidant properties in P. frutescens, suggesting its potential to boost the therapeutic properties of host plants. Graphical Abstract

Objective To investigate the serological and molecular characteristics of twenty blood samples carrying ABO*AW.37 allele and to analyze ABO allelic enhancement. Methods The ABO phenotype of the twenty samples was determined by serological methods and the genotype of 1-7 ABO exons was analyzed by Sanger sequencing. Results Sequencing analysis showed that all twenty samples contained a c. 940A>G(p.Lys314Glu) mutation of A allele, which was defined as ABO*AW.37. When ABO*AW.37 and B alleles were inherited simultaneously in 9 cases, in forward typing anti-A antibodies all agglutinated and the serological phenotype was AwB. Among the 11 cases with ABO*AW.37 and O alleles inherited simultaneously, there was no agglutination of anti-A in forward typing. For absorption and elution tests, 5 cases were weakly positive and the serological phenotype was Ael, while 6 cases were negative for absorption and elution tests and the serological phenotype was O type. Conclusion Allelic enhancement occured when both ABO*AW.37 allele and B allele were inherited simultaneously. When ABO*AW.37 was inherited simultaneously with O allele, the serological phenotype was Ael or O type and attention should be paid to blood type identification.

Long non‐coding RNAs (lncRNAs) and dendritic cells (DC) play crucial roles in the development of acute coronary syndrome (ACS); however, the mechanisms remain unclear. To investigate this, we analysed the differentially expressed lncRNAs in monocyte‐derived DCs (moDCs) from patients with ACS. Peripheral blood mononuclear cells were transformed into moDCs. Cellular morphology and expression levels of moDC‐specific markers (CD80, CD86, CD11c, CD14 and HLA‐DR) were analysed using electron microscopy (EM) and flow cytometry (FCM), respectively. Differentially expressed lncRNAs and their functions were predicted using gene sequencing, gene ontology and the Kyoto Encyclopedia of Genes and Genomes. The expression levels of markers, signalling pathway molecules (p‐PI3K and p‐AKT), inflammatory cytokines (IL‐6 and IL‐12p70) and target gene (C‐C motif chemokine ligand (CCL) 15 and CCL14) were analysed by overexpression or silencing of candidate lncRNAs. EM revealed the cells to be suspended in dendritic pseudopodia. CD11c and HLA‐DR were upregulated, while CD80 and CD86 were downregulated. Comparison between the UA versus ST group showed the highest number of differentially expressed lncRNAs (n = 113), followed by UA versus NST (n = 115), CON versus NST (n = 49) and CON versus ST (n = 35); however, the number was low for CON versus UA and ST versus NST groups. moDC‐specific marker expression, signalling pathway molecules, inflammatory cytokines and CCL14 were upregulated following lentiviral overexpression of smart silencer‐CCL15‐CCL14; however, expression levels decreased following transfection with siRNA. The morphology, function and lncRNA expression of moDCs differ depending on the type of ACS. The differentially expressed lncRNAs, particularly CCL15‐CCL14, regulate the function of moDCs. Thus, our study provides new insights regarding the role of lncRNAs in ACS and indicates the potential use of CCL15‐CCL14 as a novel diagnostic marker and therapeutic target. [ABSTRACT FROM AUTHOR]

With rapid development and mature of single‐cell measurements, single‐cell biology and pathology become an emerging discipline to understand the disease. However, it is important to address concerns raised by clinicians as to how to apply single‐cell measurements for clinical practice, translate the signals of single‐cell systems biology into determination of clinical phenotype, and predict patient response to therapies. The present Perspective proposes a new system coined as the clinical artificial intelligent single‐cell (caiSC) with the dynamic generator of clinical single‐cell informatics, artificial intelligent analyzers, molecular multimodal reference boxes, clinical inputs and outs, and AI‐based computerization. This system provides reliable and rapid information for impacting clinical diagnoses, monitoring, and prediction of the disease at the single‐cell level. The caiSC represents an important step and milestone to translate the single‐cell measurement into clinical application, assist clinicians' decision‐making, and improve the quality of medical services. There is increasing evidence to support the possibility of the caiSC proposal, since the corresponding biotechnologies associated with caiSCs are rapidly developed. Therefore, we call the special attention and efforts from various scientists and clinicians on the caiSCs and believe that the appearance of the caiSCs can shed light on the future of clinical molecular medicine. [ABSTRACT FROM AUTHOR]

Background: Endophytic fungi (EF) reside within plants without causing harm and provide benefits such as enhancing nutrients and producing bioactive compounds, which improve the medicinal properties of host plants. Selecting plants with established medicinal properties for studying EF is important, as it allows a deeper understanding of their influence. Therefore, the study aimed to investigate the impact of EF after inoculating the medicinal plant Perilla frutescens, specifically focusing on their role in enhancing medicinal properties. Results: In the current study, the impact of two EF i.e., Irpex lenis and Schizophyllum commune isolated from A. bracteosa was observed on plant Perilla frutescens leaves after inoculation. Plants were divided into four groups i.e., group A: the control group, group B: inoculated with I. lenis; group C: inoculated with S. commune and group D: inoculated with both the EF. Inoculation impact of I. lenis showed an increase in the concentration of chlorophyll a (5.32 mg/g), chlorophyll b (4.46 mg/g), total chlorophyll content (9.78 mg/g), protein (68.517 ± 0.77 mg/g), carbohydrates (137.886 ± 13.71 mg/g), and crude fiber (3.333 ± 0.37%). Furthermore, the plants inoculated with I. lenis showed the highest concentrations of P (14605 mg/kg), Mg (4964.320 mg/kg), Ca (27389.400 mg/kg), and Mn (86.883 mg/kg). The results of the phytochemical analysis also indicated an increased content of total flavonoids (2.347 mg/g), phenols (3.086 mg/g), tannins (3.902 mg/g), and alkaloids (1.037 mg/g) in the leaf extract of P. frutescens inoculated with I. lenis. Thus, overall the best results of inoculation were observed in Group B i.e. inoculated with I. lenis. GC-MS analysis of methanol leaf extract showed ten bioactive constituents, including 9-Octadecenoic acid (Z)-, methyl ester, and hexadecanoic acid, methyl ester as major constituents found in all the groups of P. frutescens leaves. The phenol (gallic acid) and flavonoids (rutin, kaempferol, and quercetin) were also observed to increase after inoculation by HPTLC analysis. The enhancement in the phytochemical content was co-related with improved anti-oxidant potential which was analyzed by DPPH (% Inhibition: 83.45 µg/ml) and FRAP (2.980 µM Fe (II) equivalent) assay as compared with the control group. Conclusion: Inoculation with I. lenis significantly enhances the uptake of nutritional constituents, phytochemicals, and antioxidant properties in P. frutescens, suggesting its potential to boost the therapeutic properties of host plants. [ABSTRACT FROM AUTHOR]

Background: Human bloodstream microRNAs (miRNAs) have emerged as a promising predictive and diagnostic biomarker for a range of cancers, including breast cancer. Our objective was to look into new miRNA biomarkers for diagnosis in the serum of patients with breast cancer and track the expression levels at different stages using miRNA profiling. Methods: 53 breast cancer patients and 10 healthy controls had blood samples tested for three miRNAs. miRNAs were extracted from blood and evaluated using real-time quantitative polymerase chain reaction. After extracting genomic DNA, miRNA primer-produced PCR products were sequenced to discover point mutations that may contribute to the illness. Results: After examining miR195, miR200b, and miR331, breast cancer patients had a significantly lower miR195 level than healthy persons. In addition, miR200b expression levels were significantly lower in breast cancer patients than in healthy individuals. In advanced stages, miR331 expression was substantially higher than in healthy people. Conclusion: The findings of our study demonstrated a significant association between the expression of miRNAs and the prognosis of breast cancer. Additional research is necessary to study better the correlation between these circulating miRNAs and the stages of breast cancer. [ABSTRACT FROM AUTHOR]

In the deep-sea environment, the volume available for an in-situ gene sequencer is severely limited. In addition, optical imaging systems are subject to real-time, large-scale defocusing problems caused by ambient temperature fluctuations and vibrational perturbations. To address these challenges, we propose an edge detection algorithm for defocused images based on grayscale gradients and establish a defocus state detection model with nanometer resolution capabilities by relying on the inherent critical illumination light field. The model has been applied to a prototype deep-sea gene sequencing microscope with a 20× objective. It has demonstrated the ability to focus within a dynamic range of ±40 μm with an accuracy of 200 nm by a single iteration within 160 ms. By increasing the number of iterations and exposures, the focusing accuracy can be refined to 78 nm within a dynamic range of ±100 μm within 1.2 s. Notably, unlike conventional photoelectric hill-climbing, this method requires no additional hardware and meets the wide dynamic range, speed, and high-accuracy autofocusing requirements of deep-sea gene sequencing in a compact form factor. [ABSTRACT FROM AUTHOR]

We report an elderly male with mucin-producing urothelial-type adenocarcinoma of the prostate (MPUAP) and oligometastatic lung involvement, initially diagnosed as benign prostatic hyperplasia and treated with transurethral plasma resection of the prostate (TURP). Postoperative pathology indicated mucinous adenocarcinoma, with immunohistochemistry positive for CK7, CK20, and CDX-2. Next-generation sequencing (NGS) identified genetic alterations similar to those found in intestinal adenocarcinoma. After ruling out gastrointestinal and bladder tumors, MPUAP was confirmed. Ablation therapy was performed for the lung metastasis, followed by radical prostate chemoradiotherapy. Post chemoradiotherapy, the patient received XELOX + Bevacizumab regmien but switched to capecitabine monotherapy due to adverse effects. At a 12-month follow-up post-radiotherapy, no prostate recurrence was observed, though previous lung nodule ablation suggested recurrence. By reviewing historical cases, we discussed the role and significance of radical resection and TURP in MPUAP. NGS is recommended for patients with MPUAP, and regarding chemotherapy, treatment options for colorectal cancer are worth considering.

The species and strain of Cryptosporidium parvum genotype in domestic ducks were recorded for the first-time using sequencing technologies in this study. In addition, it looks at the spread of Cryptosporidium parvum and the methods used to diagnose it in different districts of Mosul. Ninety-one duck feces were gathered from six different districts of the city: Shallalat, Bashiqa, Ali Rash, Bartella, Al-Hamdaniya, Telkaif, Felfil, and Al-Qosh. The positive result rate was 27% (25 of the original 91), with monthly variations based on geographic region and age. The statistical analysis revealed considerable disparities in scientific procedures between the northern and eastern Mosul regions. The age factor revealed a substantial difference between 9 and 12 months. Histopathological changes in the trachea and intestinal tract were recorded. The type and strain of Cryptosporidium parvum were recorded and compared to sequences stored in the NCBI's GenBank database. The parasite's genetic invasion, was discovered to match the strains reported in China. The study concluded that for the first time in Mosul, the strain and type of Cryptosporidium parvum isolate were recorded in local ducks, with a higher incidence among local ducks aged 6-12 months than at other ages. Furthermore, its incidence was higher in the city's northern areas than in its eastern areas. Duck trachea and intestines are affected by this strain's histopathological alterations.

Abstract Background The incidence estimates for intracranial subdural empyema are 0.1 per 100,000 individuals, making it a rare clinical condition. It is a highly morbid and fatal illness that is most commonly the result of a primary infection somewhere else. Case presentation The authors present a young male patient 15 years presented with 1 week history of headache, fever, confusion and seizure attack with initial negative CT head and nearly negative CSF analysis. With few days he developed left sided weakness. MRI brain showed right temporoparietal subdural fluid collection with midline shift. Urgent neurosurgical evacuation was made and revealed subdural empyema. The boy received combination therapy of vancomycin, ceftriaxone, and metronidazole. The culture of the pus was negative but 16S rRNA gene sequencing (bacterial) revealed streptococcus intermedius. He made a good recovery with no recollection or neurological deficit on follow up. Conclusion This case highlights the possibility of occurrence of this rare infection in otherwise healthy individuals without obvious precipitating factor. It also indicates the superiority of MRI brain over CT head in detection of subdural collection. The rapid diagnosis and intervention improve the outcome of the patient.

Objective To explore the blood group changes of two acute myeloid leukemia patients with suspected O type, and their relationship with the therapeutic effect. Methods Serological analysis of ABO blood group of patients was carried out by microcolumn gel method, tube method and absorption-elution test, ABO blood group genotyping was performed by microfluidic chip method. Exons E2 to E7 of ABO gene were amplified by PCR and sequenced by Sanger method. Results The forward typing of two cases were both O type, but the reverse typing were both A type. The absorption-elution test results all showed detection of antigen A. ABO gene phenotype of the two cases were both A, with genotyping results as A102/A102 and A102/O01, respectively. Sequencing results showed that SNP sites of ABO blood group were 467T/T, 261G/delG and 467C/T, respectively.In one case, the intensity of anti-A agglutination reaction changed significantly from weak to strong with the progress of treatment. Conclusion For clinical samples of acute myeloid leukemia patients with ABO forward and reverse typing discrepancy and suspected O type, the result of reverse typing should be valued, and absorption-elution test should be performed to further confirm the ABO blood type combining the genetic test results, so as to develop appropriate blood transfusion strategies for patients.

Objective To investigate the serological and molecular biology characteristics of ambiguous blood types among voluntary blood donors in Putian, and to file records for relevant donors for future reference in case of their own transfusion needs or emergency donations to others. Methods A total of 68 593 blood samples from voluntary blood donors in Putian Central Blood Station between January 1, 2019 and August 31, 2023 were collected and tested for blood types using serological methods. ABO gene (including promoters, enhancers and seven exons as well as their flanking sequences and intron 6) and FUT1 gene testing were performed on ambiguous blood types.3D models were constructed using Chimira and PyMOL software to predict the impact of gene mutations on enzyme structure. Results A total of 16 ABO subtypes were identified by serological methods, with the highest detection rate as the para-Bombay phenotype (0.73 per 10 000), followed by the cisAB phenotype (0.44 per 10 000). Gene analysis revealed 12 cases with known mutations (4 cases of FUT1.01N.06/FUT1.01N.06, 1 case of FUT1 01W.08/FUT1.01N.06, 2 cases of A2.08/B.01, 1 case of BA.02/O.01.01, 1 case of A3.07/O.01.01, 3 cases of cisAB.01/B.01), and key mutations were not found in 4 cases (2 cases of A1.02/B.01, 2 cases of A1.02/O.01.02). 3D molecular model analysis revealed that both A3.07 and FUT1.01W.08 allele can lead to a decrease in activity of the corresponding glycosyltransferases, resulting in the emergence of subtypes. Conclusion The most common phenotype causing discrepancies in ABO blood type testing among voluntary blood donors from Putian is the para-Bombay phenotype, with the most common allele being FUT1.01N.06.

BackgroundThe serious air pollution of highway toll booths poses a high occupational exposure risk to toll collectors. It is urgent to develop purification methods suitable for airborne particles and microbial pathogens in highway toll booths. ObjectiveTo verify the purification effect of low temperature plasma air purifiers on airborne particles and microbes in highway toll booths. MethodsBased on controlled-intervention design, we selected three toll booths in an expressway toll station as on-site experimental locations for 6 d (no-intervention period: the low-temperature plasma purifier was turned off in the first three days; intervention period: the purifier was turned on from 9:00 to 17:00 in the following three days). The indoor and outdoor PM2.5 and PM10 concentrations were continuously monitored during the study. At 9:00, 12:00, and 17:00 of every day during the experiment, indoor and outdoor air samples were collected to analyze the concentration of airborne culturable colonies with a plankton sampler. Airborne particle samples were collected in the outermost exit continuously from 9:00 to 17:00 every day during the experiment using a medium flow particulate sampler, and the species and relative abundance of fungi and bacteria contained in the samples were analyzed by gene sequencing. Independent-sample t test was used to compare the concentration of indoor PM2.5, PM10, and culturable colonies between the intervention period and the non-intervention period. α diversity analysis, β diversity analysis, and t test were used to compare the diversity and relative abundance of specific species of bacteria and fungi, as well as typical pathogenic bacteria and fungi in the samples between the non-intervention period and the intervention period to reflect the purification effect of low temperature plasma air purifier on airborne PM2.5, PM10, and microorganisms. ResultsDuring the intervention period, the mean indoor concentrations of PM2.5, PM10, and culturable colonies were lower than those of the no-intervention period (P

Nanopore sequencing is the third generation sequencing (TGS) technology based on single molecule. It has the characteristics of no amplification, easy to carry, real-time sequencing, ultra long reading length, direct sequencing of RNA, and detection of base modification. In recent years, nanopore sequencing technology has developed rapidly and has been widely applied in molecular plant breeding fields such as plant genome resequencing, whole genome, transcriptomics, organelle genomics, and epigenetic transcriptomics. The article elaborates on the research status and application prospects of nanopore sequencing technology in potato breeding at home and abroad, in order to provide new ideas for the high-quality development of China′s potato industry. [ABSTRACT FROM AUTHOR]

Background: The incidence estimates for intracranial subdural empyema are 0.1 per 100,000 individuals, making it a rare clinical condition. It is a highly morbid and fatal illness that is most commonly the result of a primary infection somewhere else. Case presentation: The authors present a young male patient 15 years presented with 1 week history of headache, fever, confusion and seizure attack with initial negative CT head and nearly negative CSF analysis. With few days he developed left sided weakness. MRI brain showed right temporoparietal subdural fluid collection with midline shift. Urgent neurosurgical evacuation was made and revealed subdural empyema. The boy received combination therapy of vancomycin, ceftriaxone, and metronidazole. The culture of the pus was negative but 16S rRNA gene sequencing (bacterial) revealed streptococcus intermedius. He made a good recovery with no recollection or neurological deficit on follow up. Conclusion: This case highlights the possibility of occurrence of this rare infection in otherwise healthy individuals without obvious precipitating factor. It also indicates the superiority of MRI brain over CT head in detection of subdural collection. The rapid diagnosis and intervention improve the outcome of the patient. [ABSTRACT FROM AUTHOR]

Neuroblastoma (NB) is the most common extra-cranial malignancy in children. Poor survival in high-risk NB is attributed to recurrent metastatic disease. To better study metastatic disease, we used a novel mouse model to investigate differential gene expression between primary tumor cells and metastatic cells. We hypothesized that metastatic NB cells have a different gene expression profile from primary tumor cells and cultured cells. Using three human NB cell lines (NGP, CHLA255, and SH-SY5Y), orthotopic xenografts were established in immunodeficient nod/scid gamma mice via subcapsular renal injection. Mice were sacrificed and NB cells were isolated from the primary tumor and from sites of metastasis (bone marrow, liver). RNA sequencing, gene set analysis, and pathway analysis were performed to identify differentially expressed genes and molecular pathways in the metastatic cells compared to primary tumor cells. There were 266 differentially expressed genes in metastatic tumor cells (bone marrow and liver combined) compared to primary tumor cells. The top upregulated gene was KCNK1 and the top downregulated genes were PDE7B and NEBL. Top upregulated pathways in the metastatic cells were involved in ion transport, cell signaling, and cell proliferation. Top downregulated pathways were involved in DNA synthesis, transcription, and cellular metabolism. In metastatic NB cells, our study identified the upregulation of biologic processes involved in cell cycle regulation, cell proliferation, migration, and invasion. Ongoing studies aim to validate downstream translation of these genomic alterations, as well as target these pathways to more effectively suppress and inhibit recurrent metastatic disease in NB. [ABSTRACT FROM AUTHOR]

Pulmonary hypertension (PH) is a progressive and potentially fatal complication of sickle cell disease (SCD), affecting 6–10% of adult SCD patients. Various mechanisms and theories have been evaluated to explain the pathophysiology of this disease. However, questions remain, particularly regarding the clinical heterogeneity of the disease in terms of symptoms, complications, and survival. Beyond the classical mechanisms that have been thoroughly investigated and include hemolysis, nitric oxide availability, endothelial disorders, thrombosis, and left heart failure, attention is currently focused on the potential role of genes involved in such processes. Potential candidate genes are investigated through next-generation sequencing, with the transforming growth factor-beta (TGF-β) pathway being the initial target. This field of research may also provide novel targets for pharmacologic agents in the future, as is already the case with idiopathic PH. The collection and processing of data and samples from multiple centers can yield reliable results that will allow a better understanding of SCD-related PH as a part of the disease's clinical spectrum. This review attempts to capture the most recent findings of studies on gene polymorphisms that have been associated with PH in SCD patients. [ABSTRACT FROM AUTHOR]

Background: Norovirus is the predominant pathogen causing foodborne illnesses and acute gastroenteritis (AGE) outbreaks worldwide, imposing a significant disease burden. This study aimed to investigate the epidemiological characteristics and genotypic diversity of norovirus outbreaks in Hongshan District, Wuhan City. Methods: A total of 463 AGE cases from 39 AGE-related outbreaks in Hongshan District between January 1, 2021, and June 30, 2023, were included in the study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to identify norovirus types GI and GII in anal swab samples from all cases. Norovirus-positive samples were sequenced and analyzed for the open reading frame (ORF) 1/ORF2 hinge region. Results: 26 norovirus infectious outbreaks were reported among 39 acute diarrheal outbreaks, including 14 outbreaks in kindergartens, 8 in elementary schools, and 4 in universities. Based on clinical symptoms and epidemiological investigations, a total of 1295 individuals were identified as having been exposed to norovirus, yielding an attack rate of 35.75 %. A higher proportion of outbreaks was observed during the winter and spring seasons (38.46 %). Additionally, norovirus-positive samples were subjected to sequencing and analysis of the open reading frame (ORF) 1/ORF2 hinge region. Genotypic data for norovirus was successfully obtained from 18 (69.23 %) of the infectious outbreaks, revealing 10 distinct recombinant genotypes. GII.4 Sydney 2012 [P31] and GII.17[P17] were the predominant strains in 2021 and 2022, GII.3 [P12] emerged as the dominant strain in 2023. Conclusion: Norovirus outbreaks in Hongshan District predominantly occurred in crowded educational institutions, with peaks in the cold season and a high attack rate in universities. GII.3 [P12] has become the locally predominant strain.

BackgroundMuscle is an important tissue of the human body. Muscle atrophy is common in people of all ages, which will lead to human weakness and decline of motor function, which is one of the important causes of disability. The common methods of genomics research are transcriptome, proteomics and metabolomics, which are important means to explore the molecular pathology of diseases. In recent years, combinatorial research has been carried out on a large scale in the field of muscle atrophy. However, no author in this field has carried out bibliometrics and visual analysis.MethodsIn this study, articles related to the histological study of muscular dystrophy since 2000 were searched from the Web of Science core database (WoSCC). We will retrieve the results through CiteSpace, VosViewer and R for data statistics and visual analysis.ResultsIn this study, a total of 141 publications were collected, and the number of publications increased year by year. These 141 articles came from 1031 co-authors from 361 institutions in 31 countries and were published in 92 journals. A total of 6286 articles from 1383 journals were cited. Authors from American institutions have published the most articles and have been cited the most, and authors from other countries have also made considerable contributions.ConclusionThis is the first bibliometric and visual analysis of published research in the field of muscular dystrophy through systematic data retrieval and combined with a variety of bibliometric analysis tools. Through these data, we summarize the previous studies of scholars, and provide prospects for future research in the field.

BackgroundTo compare the diagnostic performance of microbiological culture and 16S/18S rRNA gene polymerase chain reaction (PCR)-Sanger sequencing for infectious keratitis (IK) and to analyse the effect of clinical disease severity on test performance and inter-test concordance.MethodsThis was a three-arm, diagnostic cross-sectional study. We included all eligible patients who presented with presumed bacterial/fungal keratitis to the Queen's Medical Centre, Nottingham, UK, between June 2021 and September 2022. All patients underwent simultaneous culture (either direct or indirect culture, or both) and 16S (pan-bacterial)/18S (pan-fungal) ribosomal RNA (rRNA) PCR-Sanger sequencing. The bacterial/fungal genus and species identified on culture were confirmed using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. Relevant clinical data were also collected to analyze for any potential clinico-microbiological correlation. Main outcome measures included the diagnostic yield, test accuracy (including sensitivity and specificity), and inter-test agreement [including percent agreement and Cohen's kappa (k)].ResultsA total of 81 patients (86 episodes of IK) were included in this study. All organisms identified were of bacterial origin. Diagnostic yields were similar among direct culture (52.3%), indirect culture (50.8%), and PCR (43.1%; p = 0.13). The addition of PCR enabled a positive diagnostic yield in 3 (9.7%) direct culture-negative cases. Based on composite reference standard, direct culture had the highest sensitivity (87.5%; 95% CI, 72.4–95.3%), followed by indirect culture (85.4%; 95% CI, 71.6–93.5%) and PCR (73.5%; 95% CI, 59.0–84.6%), with 100% specificity noted in all tests. Pairwise comparisons showed substantial agreement among the three tests (percent agreement = 81.8–86.2%, Cohen's k = 0.67–0.72). Clinico-microbiological correlation demonstrated higher culture-PCR concordance in cases with greater infection severity.ConclusionsThis study highlights a similar diagnostic performance of direct culture, indirect culture and 16S rRNA PCR for bacterial keratitis, with substantial inter-test concordance. PCR serves as a useful diagnostic adjuvant to culture, particularly in culture-negative cases or those with lesser disease severity (where culture-PCR concordance is lower).

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.

Kazakhstani researchers reported a significant prevalence of highly carcinogenic human papillomavirus types in the country. The article aimed to present HPV L1 gene sequencing developments in women affected with cervical cancer throughout the western part of Kazakhstan with provided findings on the geographic pathways of obtained isolates. Methods. The HPV L1 gene was amplified using the consensus primers MY09HPV 5’-CGTCCMARRGGAWACTGATC-3’ and MY11HPV 5’ – GCMCAGGGWCATAAYAATGG-3’. The purified DNA was used as the target for direct nucleotide sequencing. Phylogenetic analyses were conducted using the MegAlign program from the LASERGENE software package (version 6.0; DNA star, Madison, WI) and with MEGA version 5.0 software. A multiple alignment was created through Clustal W software, and the neighbor-joining method was used to construct the phylogenetic tree. Results. Of 70 HPV samples transported to the Astana shared laboratory for gene L1 sequencing, only ten appeared fit to obtain isolates (14.3%). The viral load of the samples ranged from 3.3 to 8.2, and the range of DNA concentration was from 8.16 to 69.6 ng/uL. HPV16 unique Kazakhstani isolate from Aktobe, having its own branch, and not yet registered in the world genebank, was revealed. An isolate of potentially carcinogenic HPV53 forming a remote cluster with KF436822/1, KU951264.1 - Southwest China, and 97% identity with EU056643.1 - Ireland, and acted as a single agent for invasive cervical cancer was identified. In general, the sequencing findings indicate the variety of ways for HPV pervasion into the western region of Kazakhstan: North and South America, Europe, and Asia. The study was recorded in the ISRCTN registry, No. 7154910, 02/01/2018.

Panel‐based methods are commonly employed for the analysis of novel gene fusions in precision diagnostics and new drug development in cancer. However, these methods are constrained by limitations in ligation yield and the enrichment of novel gene fusions with low variant allele frequencies. In this study, we conducted a pioneering investigation into the stability of double‐stranded adapter DNA, resulting in improved ligation yield and enhanced conversion efficiency. Additionally, we implemented blocker displacement amplification, achieving a remarkable 7‐fold enrichment of novel gene fusions. Leveraging the pre‐enrichment achieved with this approach, we successfully applied it to Nanopore sequencing, enabling ultra‐fast analysis of novel gene fusions within one hour with high sensitivity. This method offers a robust and remarkably sensitive mean of analyzing novel gene fusions, promising the discovery of pivotal biomarkers that can significantly improve cancer diagnostics and the development of new therapeutic strategies. [ABSTRACT FROM AUTHOR]

As a cornerstone of the central dogma of molecular biology, RNA plays vital roles in living organisms. Over the past few decades, many RNA labeling technologies have been developed to elucidate the biological function of RNA. These technologies have significantly advanced our understanding of RNA secondary structure, localization, and turnover. Additionally, taking advantage of these innovative RNA labeling approaches, plenty of tool kits have been devised for the regulation of RNA-related biological process, such as gene expression and gene editing. In this review, we primarily focus on an array of intracellular RNA labeling methods, encompassing chemical probes-based labeling, metabolic labeling, and proximity-dependent labeling. We also provide a brief overview of their applications in the research of RNA biology. Finally, the perspectives of RNA labeling are also discussed. [ABSTRACT FROM AUTHOR]

In the long-term operation of any deep geological repository (DGR), microbial species can grow within and biological activity may take place. The presence of microbes can result in the production of small organic acids and siderophores that may change the properties of the buffer material and corrode the metal canisters. In the current study, the bacterial population of Bikaner and Barmer bentonites was investigated through the plate culture technique. For detailed analysis, 16S ribosomal RNA (rRNA) gene sequencing technique was carried out. The results showed that Bacillus tequilensis KTCT 13622(T) and Lysinibacillus fusiformis NBRC 15717(T) were present in the Bikaner bentonite, whereas Burkholderia contaminans LMG 23361(T) and Staphylococcus pasteuri ATCC 51129(T) were present in the Barmer bentonite. Bacillus tequilensis and Lysinibacillus fusiformis are rod-shaped, gram-positive, nonmoveable bacteria of phylum Firmicutes. Burkholderia contaminans is a gram-negative, rod-shaped, moveable bacterium of phylum Proteobacteria. Staphylococcus is a genus of gram-positive bacteria in phylum Firmicutes. Bacteria belonging to the phylum Firmicutes can restrict the mobility of toxic radionuclides by the process of biosorption; and bacteria belonging to the phylum Actinobacteria have the ability to change their shape by extending out branches, or filaments, and destabilizing the compactness of the backfill material. The identified bacteria would be helpful for designing DGRs, considering the long-term safety aspects. [ABSTRACT FROM AUTHOR]

Background: Pituitary stalk interruption syndrome (PSIS) is a complex clinical syndrome characterized by varied pituitary hormone deficiencies, leading to severe manifestations across multiple systems. These include lifelong infertility, short stature, mental retardation, and potentially life-threatening pituitary crises if not promptly diagnosed and treated. Despite extensive research, the precise pathogenesis of PSIS remains unclear. Currently, there are two proposed theories regarding the pathogenic mechanisms: the genetic defect theory and the perinatal injury theory. Methods: We systematically searched English databases (PubMed, Web of Science, Embase) and Chinese databases (CNKI, WanFang Med Online, Sinomed) up to February 24, 2023, to summarize studies on gene sequencing in PSIS patients. Enrichment analyses of reported mutated genes were subsequently performed using the Metascape platform. Results: Our study included 37 articles. KEGG enrichment analysis revealed mutated genes were enriched in the Notch signaling pathway, Wnt signaling pathway, and Hedgehog signaling pathway. GO enrichment analysis demonstrated mutated genes were enriched in biological processes such as embryonic development, brain development, axon development and guidance, and development of other organs. Conclusion: Based on our summary and analyses, we propose a new hypothesis: disruptions in normal embryonic development, partially stemming from the genetic background and/or specific gene mutations in individuals, may increase the likelihood of abnormal fetal deliveries, where different degrees of traction during delivery may lead to different levels of pituitary stalk interruption and posterior lobe ectopia. The clinical diversity observed in PSIS patients may result from a combination of genetic background, specific mutations, and variable degrees of traction during delivery. [ABSTRACT FROM AUTHOR]

In this study, we utilized the remarkable capabilities of Bacillus subtilis ls-45 during the fermentation process to generate pine nut peptide. Through gene sequencing, we confirmed the proficiency of Bacillus subtilis ls-45 in producing protease, thereby serving as a valuable enzymatic source for protein hydrolysis. Our investigation focused on examining the variations in amino acid types and quantities between enzymatic pine nut protein peptide (EPP) and fermented pine nut protein polypeptide (FPP). Furthermore, we conducted a comprehensive assessment of the in vitro antioxidant activities of EPP and FPP, encompassing measurements of their Hydroxyl radical scavenging rate, Total reducing capacity, Superoxide anion scavenging rate, and ABTS+ radical scavenging rate. Notably, FPP exhibited superior antioxidant capacity compared to EPP. By employing semi-inhibitory mass concentration (IC50) analysis, we determined that FPP displayed enhanced efficacy in neutralizing hazardous free radicals when compared to EPP. [ABSTRACT FROM AUTHOR]

This study investigates the evolution of the biofilm matrix responsible for treating effluent from a pulp mill and identifies the microbial community by 16S rRNA gene sequencing. In addition, a biocarrier promising a functional structure with better specific features for biofilm formation than traditional polyurethane carriers was explored. The average removal efficiencies were 43.7% for chemical oxygen demand (COD) and 62.7% for biochemical oxygen demand (BOD5). The color increased during the treatment, indicating anoxic zones being formed in the inner part of this type of carrier. Periodic micrographs showed the evolution of extracellular polymeric substances and materials like fungi and bacteria adhered to the carriers. Genetic sequencing confirmed the presence of Bacillus sp. and Paenibacillus glucanolyticus, species with the potential to degrade and discolor pulp industrial effluents. Results offer a potential basis to enhance treatment facilities of pulp and paper mills based on microbial activities.

Hundreds of bacteria, viruses, eukaryotes, and archaea comprise the human intestinal microbiota, which constitutes a complex ecology. Recent years have established the significance of the gastrointestinal microbiome in the study of biological microorganisms. Diverse techniques for cultural extraction and analysis make it difficult to cultivate intestinal gut bacteria species. To determine the causative microorganism, it is necessary to investigate microbes and bacteria. Novel and efficacious remedies for illnesses are required. Bioinformatics and next-generation sequencing can assist in the analysis of enormous quantities of sequenced data for bacterial investigations, thereby expanding sequencing. However, advancements in sequencing technology have broadened the scope of bioinformatics identification and analysis. With next-generation sequencing techniques such as metagenomic, 16S rRNA, and meta-transcriptomic sequencing, experimental data regarding the immunological response of the intestinal gut microbiome to genetic manipulation seems to be helpful in identifying the diseases. Despite significant progress over the past two decades, there is still a lack of understanding about the ecology of diseases and treatments. The present review highlights how microbial gene sequencing and analysis of sequencing information can help manage human gut issues.

Cells constitute the fundamental units of living organisms. Investigating individual differences at the single-cell level facilitates an understanding of cell differentiation, development, gene expression, and cellular characteristics, unveiling the underlying laws governing life activities in depth. In recent years, the integration of single-cell manipulation and recognition technologies into detection and sorting systems has emerged as a powerful tool for advancing single-cell research. Raman cell sorting technology has garnered attention owing to its non-labeling, non-destructive detection features and the capability to analyze samples containing water. In addition, this technology can provide live cells for subsequent genomics analysis and gene sequencing. This paper emphasizes the importance of single-cell research, describes the single-cell research methods that currently exist, including single-cell manipulation and single-cell identification techniques, and highlights the advantages of Raman spectroscopy in the field of single-cell analysis by comparing it with the fluorescence-activated cell sorting (FACS) technique. It describes various existing Raman cell sorting techniques and introduces their respective advantages and disadvantages. The above techniques were compared and analyzed, considering a variety of factors. The current bottlenecks include weak single-cell spontaneous Raman signals and the requirement for a prolonged total cell exposure time, significantly constraining Raman cell sorting technology’s detection speed, efficiency, and throughput. This paper provides an overview of current methods for enhancing weak spontaneous Raman signals and their associated advantages and disadvantages. Finally, the paper outlines the detailed information related to the Raman cell sorting technology mentioned in this paper and discusses the development trends and direction of Raman cell sorting.

Newcastle disease viruses (NDVs) possess a single-stranded, non-segmented RNA and are classified into different strains based on their level of pathogenicity. Due to slight differences in the molecular makeup of the viral genome among these strains, employing a pair of primers for molecular diagnostics becomes essential. This study aims to establish a new approach to detect potential NDV infection by developing molecular methods. This was accomplished by performing a single RT-PCR reaction utilizing a newly designed universal primer set targeting a remarkably conserved area within the viral M gene. Various tools and resources were utilized to generate a set of primers, including the NCBI database and the Geneious Inspirational Software for Biologists. Ninety-four oropharyngeal swabs were collected from 66 chickens and 28 pigeons showing signs of ND. Viral RNA was extracted from samples, and M genes were amplified using conventional RT-PCR and real-time quantitative RT-PCR (qRT-PCR), followed by genomic sequencing and bioinformatics. The designed primers exhibited good quality, as indicated by a Delta G value of less than -5. This suggests that the primers are unlikely to cause any issues during the PCR process. Moreover, the amplification of the M gene was achieved successfully in both conventional RT-PCR and RT-PCR for approximately all collected samples from chickens and pigeons. This successful amplification was further verified through genomic sequencing and subsequent sequence analysis. These findings provide confirmation that the designed universal primers can effectively identify and quantify NDVs using PCR assay.

Abstract Background 1P36 deletion syndrome is recognized as the most common terminal microdeletion syndrome in humans, characterized by early developmental delay and consequent intellectual disability, seizure disorder, and distinctive facial features. Variable deletion locations may attributed to phenotypic variability. However, the abnormal phenotypes of hematology are rarely reported in 1P36 deletion syndrome patients. Case presentation We present a case of postnatal intellectual disability accompanied by pancytopenia. Copy number variation analysis revealed a pathogenic deletion in 1p36.331p36.32 with a deletion size of 2.21 Mb. Following successful treatment with glucocorticoids, the patient was diagnosed with immuno-related hemocytopenia (IRH). Discussion The patient experienced IRH, an uncommon characteristic of 1p36 deletion syndrome. The deletion fragment of 1p36.33-p36.32, particularly the loss of GNB1 gene, has been associated with the development of pancytopenia. Genotype-phenotype correlations are valuable in identifying the genes responsible for various clinical characteristics of the syndrome by associating phenotypic variation with specific genes located within the chromosome deletion region. Genome sequencing is recommended in cases where clinical manifestations indicate the presence of a genetic disorder but pose diagnostic challenges.

We evaluated the effects of feeding 60% dried corn distillers’ grains plus solubles (DDGS) or the equivalent sulfur as CaSO4 on sperm characteristics and transcript abundance. Thirty-six half-sibling Angus bulls (256 ± 8.5 d; initial BW = 320 ± 2.7 kg) were assigned to one of three treatments: (1) 60% concentrate as corn (CON); (2) 60% DDGS as corn replacement (60DDGS); and (3) CON diet + equivalent sulfur of 60DDGS added as CaSO4 (SULF). The acrosome/cell membrane integrity, mitochondrial energy potential, oxidation status, DNA integrity, and zinc signatures were analyzed via flow cytometry. Sperm-specific gene expression was assessed via RNA sequencing. The flow cytometry data were analyzed using PROC MIXED in SAS to determine the effects of treatment. Pairwise comparisons based on edgeR were used to identify differentially expressed genes. The percentage of polarized mitochondria tended to be greater (p = 0.08) for SULF compared with CON and 60DDGS. Protamine 1 was upregulated (p < 0.01; FDR = 0.10) in 60DDGS compared to CON. Zinc signature 1 in 60DDGS and SULF was reduced (p = 0.03) compared to CON. This study suggests that feeding bulls diets containing 60% DDGS had little effect on DNA integrity and gene expression.

Background: Indoor bacterial communities may change with altitude because their major contributors, outdoor bacterial communities, vary with altitude. People’s health effects from bacteria inhalation exposure can also vary with altitude because human respiratory physiology changes with oxygen content in air. Accordingly, adjusting indoor bacterial communities may help to acclimate newcomers from low-altitude environments to ultrahigh-altitude environments. To lay the groundwork for further research, we aimed to first elucidate the bacterial communities in ultrahigh-altitude residences and the effects of altitude on these communities. We collected 187 environmental samples from residential communities at ultrahigh altitudes of 3811–4651 m in Ngari, China and sequenced bacterial 16S rRNA genes. Results: On one hand, when abundant genera in ultrahigh-altitude residences and those reported by previous studies on low-altitude residences were compared, nine genera were shared, whereas other five genera were abundant only at ultrahigh altitudes. On the other hand, when the bacterial communities of residences at different ultrahigh altitudes were further compared, the bacterial composition in indoor surface samples varied significantly with altitude. The relative abundance of five bacterial genera in indoor air samples and 10 genera and three phyla in indoor surface samples varied monotonically with altitude. Conclusions: Altitude may be a long-neglected factor that shapes residential bacterial communities and thus warrants attention.

Aims: Heme oxygenase (HO-1) affords protection against ischaemia/reperfusion (I/R) injury; however, its effects on testicular I/R injury remain poorly explored. Herein, we aimed to examine the effects of HO-1 on testicular I/R injury and elucidate the underlying mechanism. Methods: Using the TALEN technique, we knocked out the HO-1 gene from rats. In vivo: Thirty hmox+/+ and 30 hmox−/− rats were randomly assigned to six groups: sham-operated (sham), I/R (the left testicle torsion/detorsion) 0 d,I/R 1d, I/R 3d, I/R 7d and I/R 28d. In vitro: GC-1 were suffered from: control,H/R (oxygen-deprivation/reoxygenation),H/R + HO-1 siRNA,H/R + c-Jun siRNA or H/R + HO-1 siRNA + c-jun.We performed immunofluorescence and immunohistochemistry experiments to detect HO-1 nuclear translocation. Flow cytometry was used to detect cell apoptosis and analyse the cell cycle. High-resolution miRNA, mRNA sequencing, reverse transcription-quantitative PCR, and western blotting were performed to identify testicular I/R injury-related genes strongly conserved in HO-1 knockout rats. A double luciferase reporter assay was performed to verify the relationship between C-jun and miR-221/222. Main findings: In vivo, HO-1 improved the pathological damage induced by testicular I/R. In GC-1 cells, we confirmed the nuclear translocation of HO-1 and its protective effect against hypoxia/reoxygenation (H/R) damage. Accordingly, HO-1 protein itself, rather than heme metabolites, might play a key role in testicular I/R. Gene sequencing was performed to screen for miR221/222 and its downstream gene, thymocyte selection-associated high mobility group box (TOX). HO-1 increased c-Jun phosphorylation in the H/R group, knocked down c-Jun in GC-1 cells, and decreased miR-221/222 expression. Inhibition of HO-1 expression decreased the expression of c-Jun and miR-221/222, which was rescued by adding c-Jun. Dual-luciferase reporter assay confirmed the interaction between c-Jun and miR-221/222. Conclusions: HO-1 could exert a protective effect against testicular I/R via the phosphorylated c-Jun-miR-221/222-TOX pathway.

Streptomyces, the largest genus in the Streptomycetaceae family and a prolific producer of antibacterial drugs, is a saprophytic soil organism that rarely causes invasive infections. Here we report a case of necrotic pneumonia caused by Streptomyces albireticuli in a 75-year-old man who presented with progressive chest tightness and dyspnea. Streptomyces albireticuli was isolated from his bronchoalveolar lavage fluid and identified through whole-genome sequencing (WGS) and phylogenetic analysis. The patient responded satisfactorily to clarithromycin therapy. The findings of this study may enhance our vigilance in identifying visceral infections caused by Streptomyces. [ABSTRACT FROM AUTHOR]