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Bo-Wen Jiang,1,2,* Xue Ji,1,2,* Zhong-Qing Lyu,3 Bing Liang,1,2 Jian-Hang Li,3 Ling-Wei Zhu,1,2 Xue-Jun Guo,1,2 Jun Liu,1,2 Yang Sun,1,2 Yan-Jing Liu3 1Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin, People’s Republic of China; 2Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun, Jilin, People’s Republic of China; 3Third Affiliated Clinical Hospital to Changchun University of Chinese Medicine, Changchun, Jilin, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yang Sun; Yan-Jing Liu, Tel +86 431-86986933, Email sunyang10@hotmail.com; 214953156@qq.comPurpose: To elucidate the contribution of a transferable plasmid harboring the blaNDM-1 gene in an Escherichia coli clinical isolate to the spread of resistance determinants.Methods: Nine extended-spectrum β-lactamase-producing E. coli were collected from diarrhea samples from a pediatric patient and genetic linkage was investigated through enterobacteriaceae repetitive intragenic consensus polymerase chain reaction (PCR). Bacterial species were identified by 16s rRNA sequencing, susceptibility testing with the use of a BD PhoenixTM-100 Automated Microbiology System, and assessment of virulence genes by PCR. The transferability of blaNDM-1 in E. coli strain TCM3e1 was confirmed by conjugation experiments. Complete sequencing of E. coli strain TCM3e1 was determined with the PacBio and Illumina NovaSeq platforms and the characteristics were analyzed with bioinformatics software.Results: The results showed that all nine E. coli strains were the same clone. E. coli strain TCM3e1 was resistant to 12 antimicrobial agents and carried the virulence gene EAST-1. Conjugation transfer analysis showed that blaNDM-1 was carried on a self-transmissible plasmid. Two copies of the blaNDM-1 gene were present on an IncC plasmid and some resistance genes with two or three copies were located downstream of the blaNDM-1 gene and formed a tandem repeat fragment (blaDNM-1-bleo-sul1- aadA17- dfrA12).Conclusion: A transmissible plasmid harboring two copies of the blaNDM-1 gene, including clonal dispersions of the blaNDM-1 gene, was identified in clinical isolates. These findings emphasized the necessity of surveillance of the plasmid-borne blaNDM-1 to prevent dissemination.Keywords: diarrheagenic Escherichia coli, multi-drug-resistant, New Delhi metallo-β-lactamase, blaDNM-1 gene

The primary aim of this study was to analyze the evolving trends and key focal points in research on cellular metabolism of colorectal cancer (CRC). Relevant publications on cellular metabolism in CRC were sourced from the Science Citation Index Expanded within the Web of Science Core Collection database. Bibliometric analysis and visualization were conducted using VOSviewer (version 1.6.18) software and CiteSpace 6.1.R6 (64-bit) Basic. A comprehensive compilation of 4722 English-language publications, covering the period from January 1, 1991 to December 31, 2022, was carefully identified and included in the analysis. Among the authors, "Ogino, Shuji" contributed the most publications in this field, while "Giovannucci, E" garnered the highest number of citations. The journal " Cancer Research " ranked first in both publication volume and citations. Institutionally, "Shanghai Jiao Tong University" emerged as the top contributor in terms of published articles, while "Harvard University" led in citation impact. In country-based analysis, the United States held the top position in both publication output and citations, closely followed by China. The increasing recognition of the significance of cellular metabolism in CRC underscores its potential for novel therapeutic approaches aimed at improving CRC management and prognosis., Competing Interests: Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

To study antimicrobial resistance and virulence genes of Klebsiella pneumoniae and Raoultella strains isolated from captive giant pandas. Non-duplicate fecal samples were collected from 128 giant pandas during 2017-2019. All isolated microbial strains were tested for antimicrobial drug susceptibility using BD verification panels. Four extended-spectrum β-lactamase resistance genes, nine virulence genes and six capsular serotype genes were detected using PCR. 42 K. pneumoniae and nine Raoultella strains were isolated from different giant pandas. Antibiotic resistance rates were 1.9%-23.5%, except for ampicillin, and 7.8% of the isolates were multidrug-resistant to 7-10 antibiotic classes. This is the first time that a multidrug-resistant R. ornithinolytica strain has been isolated from captive giant pandas. The blaTEM, blaCTX-M, blaSHV and blaDHA genes were detected in four MDR ESBL- K. pneumoniae strains. The rmpA, iutA, ybtS, iroN and iroB genes were positively detected in 11.7% of the isolates. Capsular serotype (K2, K5, K54 and K57) genes were all detected in four K. pneumoniae strains, and one was identified as hypervirulent. This study showed that MDR ESBL- K. pneumoniae, hypervirulent K. pneumoniae, MDR R. ornithinolytica and the colistin-resistant strain may pose risks to captive giant pandas and their keepers, and that the diversity of antibiotic resistance and virulence genes in Klebsiella and Raoultella should be monitored regularly., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study aimed to characterize the antimicrobial resistance and virulence of Enterococcus from dogs and cats in Northeast China and evaluate its zoonotic risk based on a total of 469 enterococci strains from 610 samples, including 238 strains of E. faecium and 128 strains of E. faecalis. The isolation rate from police dog samples was 93.79%, pet dog samples was 69.90% and pet cat samples was 76.67%. The differences in the prevalence of E. faecalis among different hosts were statistically significant (P<0.05). The assays showed that most of the virulence genes detected were existed in E. faecalis and police dogs carried the least number of virulence genes. The correlation between enterococcal surface protein (esp) and aggregation substance (asa1) was determined. Enterococci are most resistant to tetracycline and erythromycin, 68.92% of the isolates were classified as multiple drug resistant. Significant differences (P<0.01) were found between E. faecium and E. faecalis in the resistance rates of nine antimicrobials. Four positive and four negative correlations were found between virulence genes and antimicrobial resistance. The results show that Enterococcus colonization and excretion in dogs and cats were related to animal species and living environments. Some correlation between virulence factors and antimicrobial resistance was obtained. This study confirmed the presence of strains carrying multiple virulence factors and antimicrobial resistance at the same time, suggesting a public health risk for dogs and cats as reservoirs of enterococci.

Background: The association between serum total indoxyl sulfate (tIS), and cardiovascular disease (CVD) and all-cause mortality is a matter of debate. In the current study we sought to determine the association, if any, between serum tIS, and all-cause and CVD-associated mortality in patients on maintenance hemodialysis (MHD)., Methods: A prospective cohort study was conducted involving 500 MHD patients at Dalian Municipal Central Hospital from 31 December 2014 to 31 December 2020. Serum tIS levels were measured at baseline and classified as high (≥44.16 ng/ml) or low (< 44.16 ng/ml) according to the "X-tile" program. Besides, the associations between continuous serum tIS and outcomes were also explored. Predictors were tested for colinearity using variance inflation factor analysis. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using Cox proportional hazards regression models. Restricted cubic spline model was performed to assess dose-response relationships between tIS concentration and all-cause and CVD mortality., Results: During a 58-month median follow-up period, 224 deaths (132 CVD deaths) were documented. After adjustment for potential confounders, the serum tIS level was positively associated with all-cause mortality (HR = 1.02, 95% = 1.01-1.03); however, we did not detect a significant association when tIS was a dichotomous variable. Compared with the MHD population with a serum tIS level < 44.16 ng/ml, the adjusted HR for CVD mortality among those with a serum tIS level ≥ 44.16 ng/ml was 1.76 (95% = 1.10-2.82). Furthermore, we also noted the same association when the serum tIS level was a continuous variable., Conclusion: The serum tIS level was associated with higher risk of all-cause and CVD mortality among MHD patients. Further prospective large-scale studies are required to confirm this finding., (© 2022. The Author(s).)

Purpose: To elucidate the contribution of a transferable plasmid harboring the bla NDM-1 gene in an Escherichia coli clinical isolate to the spread of resistance determinants., Methods: Nine extended-spectrum β-lactamase-producing E. coli were collected from diarrhea samples from a pediatric patient and genetic linkage was investigated through enterobacteriaceae repetitive intragenic consensus polymerase chain reaction (PCR). Bacterial species were identified by 16s rRNA sequencing, susceptibility testing with the use of a BD Phoenix TM -100 Automated Microbiology System, and assessment of virulence genes by PCR. The transferability of bla NDM-1 in E. coli strain TCM3e1 was confirmed by conjugation experiments. Complete sequencing of E. coli strain TCM3e1 was determined with the PacBio and Illumina NovaSeq platforms and the characteristics were analyzed with bioinformatics software., Results: The results showed that all nine E. coli strains were the same clone. E. coli strain TCM3e1 was resistant to 12 antimicrobial agents and carried the virulence gene EAST-1. Conjugation transfer analysis showed that bla NDM-1 was carried on a self-transmissible plasmid. Two copies of the bla NDM-1 gene were present on an IncC plasmid and some resistance genes with two or three copies were located downstream of the bla NDM-1 gene and formed a tandem repeat fragment ( bla DNM-1 - bleo-sul1- aadA17- dfrA12 )., Conclusion: A transmissible plasmid harboring two copies of the bla NDM-1 gene, including clonal dispersions of the bla NDM-1 gene, was identified in clinical isolates. These findings emphasized the necessity of surveillance of the plasmid-borne bla NDM-1 to prevent dissemination., Competing Interests: The authors declare that none have any financial or non-financial conflicts of interest associated with this study., (© 2022 Jiang et al.)

Migratory birds play an important role in the spread of multidrug-resistant (MDR) bacteria. To investigate the prevalence of MDR Escherichia coli in migratory birds in China and potential relationships with the environment, a total of 1387 samples (fecal samples, cloacal swabs, or throat swabs) were collected from migratory birds from three different river basins in China. The collected samples were processed and subjected to bacteriological examinations. Antimicrobial susceptibility testing of the recovered isolates was performed using the E-test for the detection of minimum inhibitory concentrations (MICs). Some antibiotic resistance genes were detected and the PCR products were confirmed by sequencing. In total, 478 (34.7%) E. coli isolates were recovered. The results showed that the drug-resistant E. coli isolates were highly resistant to β-lactams (43.7%) and tetracycline (22.6%), and 73 (15.3%) were MDR, including eight that were extended spectrum β-lactamase-positive. The retrieved strains harbored the blaCTX-M, blaTEM-1, tet(A), tet(B), tet(M), sul1, sul2, sul3, cmlA, floR, and intI1 genes with a prevalence of 5.9%, 36.4%, 80.5%, 11.9%, 6.8%, 6.8%, 47.5%, 12.7%, 50.8%, 37.3%, and 61.0%, respectively. The drug resistance rate of the isolates from southern China was higher than those from northern China. The E. coli samples collected for migratory birds in the Pearl River Basin had the highest proportion (46.7%) MDR isolates. Furthermore, MDR bacteria carried by migratory birds were closely related to the antibiotic content in the basin, which confirms that MDR bacteria carried by migratory birds are likely acquired from the environment. This study also confirmed that migratory birds are potential transmitters of MDR bacteria, demonstrating the need to reduce the use and emission of antibiotics and further in-depth studies on the mechanisms underlying drug resistance of bacteria., Competing Interests: The authors have declared that no competing interests exist.

Herein, a metal-free and solvent-free protocol was developed for the C-N coupling of heteroaryl halides and amines, which afforded numerous heteroaryl amines or their hydrochlorides without any external base. Further investigations elucidated that the basicity of amines and specific interactions derived from the X-ray crystallography analysis of 3j'·HCl played pivotal roles in the reactions. Moreover, this protocol was scalable to gram scales and applicable to drug molecules, which demonstrated its practical value for further applications.

Psoriasis is considered an immune-mediated inflammatory skin disorder that affects the quality of life of nearly four percent of the world population. Considering the side effects of existing therapeutic drugs and the urgent need for new drug development, we screened more than 250 traditional Chinese medicine compounds to identify drugs that significantly reduced the viability of human HaCaT keratinocytes, a psoriasis-related model cell line. Convallatoxin (CNT) was found to be a highly effective inhibitor of HaCaT cell viability. Subsequent mechanistic studies revealed that CNT induced HaCaT cell death by necroptosis rather than by apoptosis. CNT destroyed the membrane integrity of HaCaT cells, as detected by nuclear propidium iodide (PI) staining and lactate dehydrogenase (LDH) release. Additionally, the intercellular levels of adenosine triphosphate (ATP) were lower in HaCaT cells treated with CNT than in control HaCaT cells, and typical necroptosis-associated characteristics were observed by electron microscopy in cells treated with CNT. Furthermore, compared with control HaCaT cells, CNT-treated HaCaT cells produced more reactive oxygen species (ROS), but this effect was inhibited by the antioxidants N-acetyl-cysteine (NAC), diphenyleneiodonium chloride (DPI), and apocynin and the necroptosis inhibitor Nec-1. In addition, antioxidant treatment attenuated necroptotic cell death, suggesting that CNT-induced HaCaT necroptosis is mediated by oxidative stress. More importantly, CNT ameliorated skin lesions and inflammation in imiquimod (IMQ)- and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced psoriasis-like mouse models. In conclusion, our results demonstrate that CNT is cytotoxic against HaCaT cells in vitro and exerts antipsoriatic activities in two mouse models of psoriasis in vivo, making CNT a potential promising candidate drug for future research., (Copyright © 2019 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)

The possible presence of graphene in the interstellar medium (ISM) is examined by comparing the interstellar extinction curve with the ultraviolet absorption of graphene calculated from its dielectric functions experimentally obtained with the electron energy loss spectroscopy (EELS) method. Based on the absence in the interstellar extinction curve of the [Formula: see text] π-π* electronic interband transition of graphene, we place an upper limit of [Formula: see text] of C/H on the interstellar graphene abundance, exceeding the previous estimate by a factor of [Formula: see text]3 which made use of the dielectric functions measured with the spectroscopic ellipsometry (SE) method. Compared with the SE method which measures graphene in air (and hence its surface is contaminated) in a limited energy range of [Formula: see text]0.7-5 [Formula: see text], the EELS probes a much wider energy range of [Formula: see text]0-50 [Formula: see text] and is free of contamination. The fact that the EELS dielectric functions are substantially smaller than that of SE naturally explains why a higher upper limit on the graphene abundance is derived with EELS. Inspired by the possible detection of C 24 , a planar graphene sheet, in several Galactic and extragalactic planetary nebulae, we also examine the possible presence of C 24 in the diffuse ISM by comparing the model IR emission of C 24 with the observed IR emission of the Galactic cirrus and the diffuse ISM towards l  = 44°20' and b  = -0°20'. An upper limit of [Formula: see text] on C 24 is also derived from the absence of the characteristic vibrational bands of C 24 at [Formula: see text]6.6, 9.8, and 20 [Formula: see text] in the observed IR emission., (© 2019 The Author(s) Published by Oxford University Press on behalf of the Royal Astronomical Society.)

Psoriasis is a common dermatosis causing considerable inconvenience to 4% of the general population. Traditional psoriasis treatments often cause side effects, drug resistance and complications, necessitating development of safer and more effective treatments. In this study, we screened over 600 natural compounds to identify viability inhibitors of human HaCaT keratinocytes cultured in vitro. The results showed that nitidine chloride was a highly effective inhibitor. Further studies revealed that nitidine chloride inhibited HaCaT proliferation and induced S phase cell cycle arrest; these effects were associated with reduced DNA synthesis, decreased Ki67, cyclin A, and cyclin D1 levels, and increased p53 protein expression. Nitidine chloride also significantly downregulated bcl-2 and upregulated bax, cleaved caspase-9 and cleaved caspase-3. Mechanistic studies revealed that nitidine chloride-induced apoptosis involved the c-Jun N-terminal kinase (JNK) pathway. More importantly, in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)- and imiquimod (IMQ)-induced epidermal hyperplasia and inflammation models, nitidine chloride inhibited topical edema in mouse ear and back skin, substantially reducing tissue thickness and weight. In some cases, nitidine chloride also ameliorated conditions caused by TPA and IMQ, such as angiogenesis and infiltration of large numbers of inflammatory cells around blood vessels. Additionally, nitidine chloride inhibited the expression of various proinflammatory cytokines in the two animal models. In conclusion, our results are the first to demonstrate that nitidine chloride inhibits the proliferation of HaCaT cells, induces apoptosis partly via the JNK signaling pathway in vitro and ameliorates skin lesions and inflammation in vivo, making it an appropriate candidate for psoriasis treatment., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study was carried out to optimize the solvent for extracting the antioxidants and α-glucosidase inhibitors (AGIs) from Pyracantha fortuneana fruit (PFF) and the major chemical components were characterized by HPLC-QTOF-MS/MS. The results showed that 50% and 70% acetone (v/v, ml/ml) gave the best extraction efficiency on phenolics and total flavonoids, while 70% acetone and 50% methanol possess better recovery on protein and polysaccharides, respectively. In addition, the 50% and 70% acetone extracts gave the strongest radical scavenging ability and α-glucosidase inhibitory activity (p > 0.05), but the Fe 3+ reducing power of the 50% acetone extract was higher than that of 70% acetone. Correlation analysis indicated that phenolic acids and flavonoids were connected to the antioxidant activity and α-glucosidase inhibitory activity closely. Moreover, 25 compounds including 7 flavonoids, 6 phenolic acids, 7 organic acids, 3 tannins, 1 terpene, and 1 alkaloid were identified or tentatively identified in the 50% acetone extract. Overall, 50% acetone can be a proper solvent for extracting antioxidants and AGIs from PFF. PRACTICAL APPLICATIONS: Imbalance between production and clearance of reactive oxygen species (ROS) in the body could induce various chronic diseases. PFF is an edible fruit beneficial to human health; it is reported to be capable of optimizing blood glucose levels and may prevent premature aging. In the present study, PFF was found to be excellent in antioxidant activities and α-glucosidase inhibitory ability; 50% acetone was found to be the best extraction solvent. In addition, the predominant phytochemical components of the 50% acetone extract were characterized. This study can promote further research of Pyracantha fortuneana in natural functional products, especially in the prevention of type II diabetes and its complication., (© 2019 Wiley Periodicals, Inc.)

As an important part of epigenetic marker, DNA methylation involves in the gene regulation and attracts a wide spread attention in biological auxology, geratology and oncology fields. In forensic science, because of the relative stable, heritable, abundant, and age-related characteristics, DNA methylation is considered to be a useful complement to the classic genetic markers for age-prediction, tissue-identification, and monozygotic twins' discrimination. Various methods for DNA methylation detection have been validated based on methylation sensitive restriction endonuclease, bisulfite modification and methylation-CpG binding protein. In recent years, it is reported that the third generation sequencing method can be used to detect DNA methylation. This paper aims to make a review on the detection method of DNA methylation and its applications in forensic science., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Forensic Medicine.)

Vaginal swabs taken in rape cases usually contain epithelial cells from the victim and sperm from the assailant and forensic DNA analysis requires separation of sperm from these cell mixtures. PH-20, which is a glycosylphosphatidylinositol-anchored hyaluronidase located on the head of sperm, has important functions in fertilization. Here we describe a newly developed method for sperm isolation using anti-PH-20 antibody-coupled immunomagnetic beads (anti-PH-20 IMBs). Optical microscopy and scanning electron microscopy showed the IMBs recognized the head of sperm specifically and exhibited a great capacity to capture sperm cells. However, we found it necessary to incubate the IMB-sperm complex with DNase I before sperm lysis in order to remove any female DNA completely. We compared the sensitivity of anti-PH-20 IMBs in sperm and epithelial cell discrimination to those coated with a different anti-sperm antibody (anti-SP-10, anti-ADAM2 or anti-JLP). Only the anti-PH-20 IMBs succeeded in isolating sperm from cell mixtures at a sperm/epithelial cell ratio of 103:105. Further, our method exhibited greater power and better stability for sperm isolation compared to the traditional differential lysis strategy. Taken together, the anti-PH-20 IMB method described here could be effective for the isolation of sperm needed to obtain a single-sourced DNA profile as an aid to identifying the perpetrator in sexual assault cases.

A combined shading system (CSS) consisting of a beam stop and a pinhole is proposed to be used between an ellipsoidal monocapillary (EM) and a conventional laboratory x-ray source to obtain an adjustable hollow-cone output beam for different experiments with no need for changing the EM. The CSS can change the incident x-ray beam on the EM by adjusting the position of the beam stop and the pinhole, with the corresponding change of the output beam of the EM. In this study, the adjustable hollow-cone output x-ray beam of an 80-mm-long EM with a CSS was studied in detail with a laboratory Cu x-ray generator with a focal spot diameter of 50 μm. The adjustable range of the focal spot size of the EM was from 8.6 to 58.7 μm. The adjustable range of the gain of the focal spot of the EM was from 0 to 1350. The beam divergence of the hollow-cone output beam of the EM ranged from 6 to 16.75 mrad. The illumination angle of the hollow-cone output beam of the EM ranged from 0 to 5.95 mrad. In addition, the potential application of the proposed adjusting method in testing the performance of the EM is briefly discussed.

Short tandem repeat (STR) genotyping methods are widely used for human identity testing applications, including forensic DNA analysis. Samples of DNA containing the length-variant STR alleles are typically separated and genotyped by comparison to an allelic ladder. Here, we describe a newly devised library of cloned STR alleles. The library covers alleles X and Y for the sex-determining locus Amelogenin and 259 other alleles for 22 autosomal STR loci (TPOX, D3S1358, FGA, D5S818, CSF1PO, D7S820, D8S1179, TH01, vWA, D13S317, D16S539, D18S51, D21S11, D2S1338, D6S1043, D12S391, Penta E, D19S433, D11S4463, D17S974, D3S4529 and D12ATA63). New primers were designed for all these loci to construct recombinant plasmids so that the library retains core repeat elements of STR as well as 5'- and 3'-flanking sequences of ∼500 base pairs. Since amplicons of commercial STR genotyping kits and systems developed in laboratories are usually distributed from 50 to <500 base pairs, this library could provide universal templates for allelic ladder preparation. We prepared three different sets of allelic ladders for this locus TH01 and an updated version of an allelic ladder for the DNATyper(®)19 multiplex system using these plasmids to confirm the suitability of the library as a good source for allelic ladder preparation. Importantly, the authenticity of each construct was confirmed by bidirectional nucleotide sequencing and we report the repeat structures of the 259 STR alleles. The sequencing results showed all repeat structures we obtained for TPOX, CSF1PO, D7S820, TH01, D16S539, D18S51 and Penta E were the same as reported. However, we identified 102 unreported repeat structures from the other 15 STR loci, supplementing our current knowledge of repeat structures and leading to further understanding of these widely used loci., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

This study was purposed to construct the recombinant hF9 minigene and its stable nonsense mutant cell lines, and to investigate its significance. Minigene hF9 was cloned into the mammalian expression vector pCMV-Tag3B; a nonsense mutant containing a premature termination codon (PTC) in the 121(st) amino acid residue was obtained by PCR site-directed mutagenesis; minigene hF9 and nonsense mutant were respectively transfected into HepG2 cells with G418 treatment to get stable HepG2-WT and HepG2-N cell lines. The results confirmed that the minigene hF9 and nonsense mutant were constructed successfully. The gene of interest was amplified by RT-PCR from the stable cell lines, and the minigene hF9 was expressed in the stable cell lines. It is concluded that the recombinant hF9 minigene and its stable nonsense mutant cell lines are constructed successfully. The cell lines can be used to screen the drugs treating the nonsense mutation-caused hemophilia according to PTC read-through approaches.