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Shan-Shan Su,1,* Sheng-Nan Zhang,1,* Jun-Ru Ye,1 Ling-Na Xu,1 Peng-Cheng Lin,1 Han-Yan Xu,1 Qing Wu,2 Yu-Ping Li1 1Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325015, People’s Republic of China; 2The Center of Laboratory and Diagnosis, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325015, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu-Ping LiDepartment of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, South Baixiang, Ouhai District, Wenzhou, Zhejiang Province 325015, People’s Republic of ChinaTel/fax +86 577 5557 9273Email wzliyp@163.comBackground: Patients with high-titer anti-IFN-γ autoantibodies present disseminated non-tuberculous mycobacterial (NTM) and other opportunistic infections. Due to its rare occurrence and non-specific symptoms, this syndrome is difficult to diagnose during early disease stages. Here, we report a case with high-concentrations of serum anti-IFN-γ autoantibodies who presented with disseminated Talaromyces marneffei and NTM disease accompanied Sweet’s syndrome.Case presentation: A 62-year-old Chinese woman with no previous history was admitted to our hospital in August 2016 due to intermittent fever for 2 years, left chest wall redness, and swelling for 3 months. During hospitalization, the patient was confirmed with disseminated T. marneffei and successfully treated with antifungal therapy. In July 2017, upon second admission, Mycobacterium avium intracellular (MAC) pulmonary infection was established after positive cultures from the right lung tissue. The patient failed treatment after 1 month of anti-NTM therapy due to side effects. In May 2018, she was confirmed as having disseminated MAC disease accompanied by hand rashes, which was considered as Sweet’s syndrome. High-level anti-IFN-γ antibodies in the patient serum were detected upon comparison with normal controls (2.85-fold increase). Following anti-NTM therapy, both symptoms and pulmonary infiltration gradually improved, and joint destruction and lymphadenitis remained.Conclusions: Patients with anti-interferon-γ autoantibodies should be considered for severe, recurrent infections in adults in the absence of other known risk factors. Sweet’s syndrome is a common skin manifestation of the syndrome.Keywords: Talaromyces marneffei, Mycobacterium avium, Sweet’s syndrome, autoantibodies, interferon-gamma

Ying Zhou,1,* Peng-Cheng Lin,1,* Jun-Ru Ye,1 Shan-Shan Su,1 Li Dong,1 Qing Wu,2 Han-Yan Xu,1 Yu-Peng Xie,1 Yu-Ping Li1 1Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, Zhejiang, China; 2The Center of Laboratory and Diagnosis, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, Zhejiang, China *These authors contributed equally to this work Background: Clinicians may fail to make an early diagnosis of pulmonary cryptococcosis (PC) without HIV infection. Serum cryptococcal capsular polysaccharide antigen (CrAg) test, histopathology and culture of lung tissue play different roles in diagnosis of PC. Objective: To investigate the performance of serum CrAg test, histopathology and culture of the lung tissue in diagnosis of PC without HIV infection. Patients/methods: From January 2011 to September 2017, patients with proven PC were recruited from a teaching hospital in southern China. Those patients with HIV infection, PC confirmed by surgery or PC with probable or possible diagnosis were excluded from the study. Latex agglutination test and CrAg lateral flow assay were used for detection of serum CrAg. Lung biopsy and needle aspiration were performed under computed tomography guidance. Results: Eighty-nine patients with proven PC including 41 male (46.1%) and 48 female (53.9%) were enrolled. Fifty-one (57.3%) patients had underlying disease. Positive CrAg test was found in 83 (93.3%) cases. Among six cases with negative CrAg test, PC was confirmed by histology in two cases and positive culture in four cases. The histopathological results of 77 (86.5%) cases revealed cryptococcal granuloma and 12 cases showed chronic inflammation, which was confirmed by positive culture. Among 65 cases, the diseased tissue of 46 (70.8%) cases presented Cryptococcus neoformans in the culture and one case was diagnosed with lung cancer coexisting with PC. Conclusion: Our findings showed that serum CrAg test is rapid and sensitive in diagnosing PC, histology is important for confirming PC and culture plays a complementary role. Biopsied lung tissue should be submitted for cultures whenever feasible. Keywords: pulmonary cryptococcosis, fungal infection, Cryptococcus neoformans, cryptococcal capsular polysaccharide antigen, diagnosis

Zhi-Hui He,1 Yan Chen,2 Ping Chen,2 Sheng-Dong He,2 Hui-Hui Zeng,2 Ji-Ru Ye,2 Da Liu,2 Jun Cao3 1Intensive Care Unit, 2Department of Respiratory Medicine, Second Xiangya Hospital, Central South University, Changsha, 3Department of Respiratory Medicine, Hunan Provincial People’s Hospital, Changsha, China Background: There is a growing realization that COPD, or at least emphysema, involves several processes presenting in aging and cellular senescence. Endothelial progenitor cells (EPCs) contribute to neovascularization and play an important role in the development of COPD. The gene for p16Ink4a is a major dominant senescence one. The aim of the present study was to observe changes in lung function, histomorphology of lung tissue, and expression of p16Ink4a in lung tissue and bone marrow-derived EPCs in emphysematous mice induced by cigarette-smoke extract (CSE), and further to search for a potential candidate agent protecting against emphysema induced by CSE. Materials and methods: An animal emphysema model was induced by intraperitoneal injection of CSE. 5-Aza-2'-deoxycytidine (5-Aza-CdR) was administered to the emphysematous mice. Lung function and histomorphology of lung tissue were measured. The p16Ink4a protein and mRNA in EPCs and lung tissues were detected using Western blotting and quantitative reverse-transcription polymerase chain reaction, respectively. Results: CSE induced emphysema with increased p16Ink4a expression in lung tissue and bone marrow-derived EPCs. 5-Aza-CdR partly protected against emphysema, especially in the lung-morphology profile, and partly protest against the overexpression of p16Ink4a in EPCs and lung tissue induced by CSE. Conclusion: 5-Aza-CdR partly protected against emphysema in mice via suppressing p16Ink4a expression in EPCs and lung tissue. Keywords: 5-Aza-2'-deoxycytidine, cigarette smoke, emphysema, endothelial progenitor cells, p16Ink4a

Efferocytosis is crucial for the clearance of apoptotic cells (ACs) following acute ischemic stroke (AIS), however, its mechanism remains unclear. This study reveals that chemokine-like factor 1 (CKLF1) disrupts efferocytosis by promoting AC finding and internalization while impairing AC degradation in microglia. CKLF1 deficiency reduced the proportion of ACs and lowered levels of damage-associated molecular patterns. Mechanistically, CKLF1 binds to phosphatidylserine on apoptotic neurons/blebs, recruiting microglia to the ischemic penumbra via a C-C chemokine receptor 4 (CCR4)-dependent pathway. Apoptotic blebs with CKLF1 are engulfed into microglia, triggering the rapid production of interleukin-6 (IL6). IL6 enhances AC internalization through the signal transducer and activator of transcription 3 (STAT3)-vav guanine nucleotide exchange factor 1 (VAV1)-ras-related C3 botulinum toxin substrate 1 (RAC1) signaling cascade but simultaneously inhibits transcription factor EB (TFEB) nuclear translocation, leading to lysosomal dysfunction. This effect results in AC accumulation, compromising microglial efferocytosis efficiency and integrity. These findings uncover a novel regulatory axis induced by CKLF1, emphasizing the complex balance between AC internalization and degradation in microglial efferocytosis., Competing Interests: Competing interests: The authors declare no competing interests. Ethics approval and consent to participate: The study was conducted in accordance with the principles of the Declaration of Helsinki principles. The animal protocols used were approved by the Institutional Animal Care and Use Committee of Peking Union Medical College and the Chinese Academy of Medical Sciences (No. 00004538)., (© 2025. The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare.)

Background: This study aimed to investigate the effects of general intravenous anesthesia and combined inhalation anesthesia on regional saturation of oxygen (rSO 2 ) and cerebral hemodynamics during carotid endarterectomy (CEA). Optimizing intraoperative brain protection strategies has become a key focus in CEA research., Methods: Fifty-four patients (43 males, 11 females, aged 44-80) undergoing unilateral CEA were randomly assigned to Group IVA (intravenous anesthesia) or Group CIA (combined inhalation anesthesia), with 27 patients each. Group IVA was maintained with propofol and remifentanil, while Group CIA used sevoflurane, propofol, and remifentanil, with sevoflurane stopped after carotid exposure. Hemodynamics were controlled at various stages: ±10% before clamping, + 20% during clamping (metaraminol), and 0 to -10% after exposure. HR, MAP, and rSO₂ were recorded at T0 (pre-induction), T1 (pre-clamping), T2 (post-clamping), T3 (5 min post-clamping), T4 (10 min post-clamping), T5 (15 min post-clamping), and T6 (15 min post-reperfusion). Blood samples were taken at T1, T6, and T7 (24 h post-surgery) for blood gas and S100-β analysis., Results: No significant differences in rSO₂ were observed at T0 and T6 (P > 0.05). However, Group CIA had significantly higher rScO₂ at T1, T2, T3, T4, and T5 (P < 0.05). From T2 to T5, rSO₂ increased in both groups (P < 0.05). MAP and HR showed no significant differences (P > 0.05). ΔrSO₂ increased more in Group CIA (P < 0.05). At T6, S100-β protein was higher in Group IVA (P = 0.016), and pH differed significantly at T1 (P = 0.009). No other significant differences were observed., Conclusion: Both intravenous and combined inhalation anesthesia may reduce rSO₂ decline during temporary clamping in CEA. Combined inhalation anesthesia showed a trend toward higher rSO₂ levels, potentially leading to better outcomes, but further studies are needed to confirm these findings., Retrospectively Registered Clinical Trial Number: ISRCTN17014575; Registration Date: 2024/6/10., Competing Interests: Declarations. Ethics approval and consent to participate: The study was approved by the Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University, and informed consent was obtained from all subjects(Clinical Trial Number: ISRCTN17014575; Registration Date: 2024/6/10, https://www.isrctn.com/ISRCTN17014575?q=ISRCTN17014575&filters=&sort=&offset=1&totalResults=1&page=1&pageSize=10 ). The final version of the experimental protocol, informed consent form, researcher manual, and Clinical Trial Observation Form (CRF) were developed and revised in accordance with the guidelines of the Ethics Committee. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Bilateral mastectomy and bilateral salpingo-oophorectomy (BSO) reduce cancer risks in BRCA mutation carriers. However, limited data exist on complication rates for mastectomy combined with BSO or with immediate breast reconstruction (IBR)., Methods: We analyzed 83,885 female patients in NSQIP (2007-2019) undergoing mastectomy, excluding partial mastectomies. Patients were grouped by surgery type: (a) mastectomy alone, (b) mastectomy with BSO, (c) mastectomy with IBR, and (d) all three procedures. Outcomes and predictors of postoperative complications were assessed., Results: Increased postoperative complications were associated with higher body mass index (BMI; OR 1.89, p < 0.001), diabetes (OR 1.76, p < 0.001), tobacco use (OR 1.27, p < 0.001), and higher ASA class (OR 1.72, p < 0.001). Complication rates were highest for patients undergoing mastectomy, IBR, and BSO (6.50% versus 5.76% for mastectomy alone, p = 0.0055). On multivariate analysis, only mastectomy with IBR was significantly associated with increased complications (OR 1.11, p < 0.001). Mastectomy with BSO had the lowest complication rate (5.39%, p = 0.0055) and showed no significant differences in (length of stay) LOS, readmission, or reoperation rates compared with mastectomy alone., Conclusions: Adding BSO to mastectomy + IBR did not significantly increase reoperation rates, supporting the safety of combined procedures. However, IBR increased complication risk and reoperation rates, emphasizing the need for preoperative risk stratification and patient counseling., Competing Interests: Disclosure: Dr. Edmonson: advisory board, Impedimed. All other authors report no financial or commercial relationships that have a special interest in this study and received no financial or material support in conducting this work. The authors declare that none among them possess any conflicts of interest, including relevant financial interests, activities, relationships, and affiliations, that would in any way affect this manuscript or its conclusions. Data Access, Responsibility, and Analysis: The first and second authors had full access to all the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. Data Sharing Statement: The database utilized in this study is a free, publicly available database from the American College of Surgeons which is available upon request. No additional non-public data were used in this study. Artificial Intelligence Use Statement: This manuscript, along with the data collection, preparation, analysis, conclusions, writing, and references, was produced solely by the authors without the use of any artificial intelligence technology., (© 2025. Society of Surgical Oncology.)

Purpose: Three-dimensional (3D) printing must go through a post-processing procedure. This study aimed to evaluate the effects of different cleaning methods on 3D printed three-unit fixed partial dentures using 3D analysis., Materials and Methods: A three-unit fixed partial denture was designed and printed using two different printers. The evaluation was taken regarding cleaning method with ethanol (ETH) or isopropanol (IPA), and combining ultrasonic (US) technique for the following groups: group 1, ETH; group 2, ETH+US; group 3, IPA; group 4, IPA+US; and group 5, US. The 3D evaluation was conducted by calculating the mean distance between two specific points and analyzing various angular deviations, utilizing the geometry-embedded library. In addition, visual analysis was performed by creating sectional planes in different points of view. Normality test and independent t-test were performed to compare results between the two printers used in this study. In addition, one-way ANOVA was performed to compare cleaning method groups., Results: Mean distance results showed statistical differences between printers for groups IPA, US, and IPA+US, whereas no significant differences were found among cleaning methods. Angular measurements were taken at the bucco-lingual, proximal, and occlusal planes to assess deviations in the internal surfaces. Overall results showed a tendency of shrinkage of the printed three-unit fixed partial denture., Conclusion: Ultrasonic treatment may impact the washing efficiency when used with other cleaning solutions regarding angular and longitudinal deformation. Adequate selection of post-processing methods is crucial to decrease morphological deformities in 3D printed materials., (© 2025 The Korean Academy of Prosthodontics.)

Emerging evidence shows that psychological stress promotes the progression of Parkinson's disease (PD) and the onset of dyskinesia in non-PD individuals, highlighting a potential avenue for therapeutic intervention. We previously reported that chronic restraint-induced psychological stress precipitated the onset of parkinsonism in 10-month-old transgenic mice expressing mutant human α-synuclein (αSyn) (hαSyn A53T). We refer to these as chronic stress-genetic susceptibility (CSGS) PD model mice. In this study we investigated whether ginsenoside Rg1, a principal compound in ginseng notable for soothing the mind, could alleviate PD deterioration induced by psychological stress. Ten-month-old transgenic hαSyn A53T mice were subjected to 4 weeks' restraint stress to simulate chronic stress conditions that worsen PD, meanwhile the mice were treated with Rg1 (40 mg· kg -1 ·d -1 , i.g.), and followed by functional magnetic resonance imaging (fMRI) and a variety of neurobehavioral tests. We showed that treatment with Rg1 significantly alleviated both motor and non-motor symptoms associated with PD. Functional MRI revealed that Rg1 treatment enhanced connectivity between brain regions implicated in PD, and in vivo multi-channel electrophysiological assay showed improvements in dyskinesia-related electrical activity. In addition, Rg1 treatment significantly attenuated the degeneration of dopaminergic neurons and reduced the pathological aggregation of αSyn in the striatum and SNc. We revealed that Rg1 treatment selectively reduced the level of the stress-sensitive protein RTP801 in SNc under chronic stress conditions, without impacting the acute stress response. HPLC-MS/MS analysis coupled with site-directed mutation showed that Rg1 promoted the ubiquitination and subsequent degradation of RTP801 at residues K188 and K218, a process mediated by the Parkin RING2 domain. Utilizing αSyn A53T + ; RTP801 -/- mice, we confirmed the critical role of RTP801 in stress-aggravated PD and its necessity for Rg1's protective effects. Moreover, Rg1 alleviated obstacles in αSyn autophagic degradation by ameliorating the RTP801-TXNIP-mediated deficiency of ATP13A2. Collectively, our results suggest that ginsenoside Rg1 holds promise as a therapeutic choice for treating PD-sensitive individuals who especially experience high levels of stress and self-imposed expectations., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Esculetin (ESC) is a coumarin-derived phytochemical prevalent in traditional Chinese medicine that exhibits anti-acute ischemic stroke activities. Our previous studies demonstrate that CKLF1 is a potential anti-stroke target for coumarin-derived compound. In this study we investigated whether CKLF1 was involved in the neuroprotective effects of ESC against photothrombotic stroke in mice. The mice were treated with ESC (20, 40 or 80 mg·kg -1 ·d -1 , i.g.) for two weeks. The therapeutic effect of ESC was assessed using MRI, neurological function evaluation, and a range of behavioral tests on D1, 3, 7 and 14 of ESC administration. We showed that oral administration of ESC dose-dependently reduced the cerebral infarction volume within one week after stroke, improved behavioral performance, and alleviated neuropathological damage within two weeks. Functional MRI revealed that ESC significantly enhanced the abnormal low-frequency fluctuation (ALFF) value of the motor cortex and promoted functional connectivity between the supplementary motor area (SMA) and multiple brain regions. We demonstrated that ESC significantly reduced the protein levels of CKLF1 and CCR5, as well as the CKLF1/CCR5 protein complex in the peri-infarcted area. We showed that ESC (0.1-10 μM) dose-dependently blocked CKLF1-induced chemotactic movement of neutrophils in the Transwell assay, reducing the interaction of CKLF1/CCR5 on the surface of neutrophils, thereby reducing neutrophil infiltration, and decreasing the expression of ICAM-1, VCAM-1 and MMP-9 in the peri-infarct tissue. Knockout of CKLF1 reduced brain infarction volume and motor dysfunction after stroke but also negated the anti-stroke efficacy and neutrophil infiltration of ESC. These results suggest that the efficacy of ESC in promoting post-stroke neural repair depends on its inhibition on CKLF1-mediated neutrophil infiltration, which offering novel perspectives for elucidating the therapeutic properties of coumarins., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Purpose: The aim of this study was to evaluate the effects of Frondoside A (FA) on the osteogenic differentiation of human periodontal ligament (PDL) cells., Methods: Human PDL cells were cultured in osteogenic medium and treated with FA at concentrations of 0, 0.05, and 0.2 µM for 14 days. The expression levels of genes associated with osteogenic differentiation were assessed using quantitative real-time polymerase chain reaction analysis. Subsequently, RNA sequencing was performed to identify enriched gene sets following FA treatment. Alkaline phosphatase (ALP) activity was measured to confirm the osteogenic potential of FA., Results: Treatment with 0.2 µM FA significantly increased the expression levels of runt-related transcription factor 2 ( RUNX2 ), ALP , and osteocalcin ( OCN ) at day 3, while also significantly elevating the expression of dentin sialophosphoprotein ( DSPP ), RUNX2 , ALP , OCN , and osterix ( OSX ) at day 14 ( P <0.017). Hallmark gene sets enriched during FA treatment were associated with the KRAS (normalized enrichment score [NES]=2.02, Q =0.000), interferon alpha (IFN-α) (NES=1.88, Q =0.001), IFN-γ (NES=1.85, Q <0.001), hypoxia (NES=1.79, Q =0.001), and p53 (NES=1.77, Q =0.001) signaling pathways. Additionally, treatment with 0.2 µM FA significantly intensified ALP staining at day 14 ( P <0.05)., Conclusions: Within the limitations of this study, FA treatment influenced periodontal regeneration by promoting the osteogenic differentiation of human PDL cells., Competing Interests: No potential conflict of interest relevant to this article was reported., (Copyright © 2024. Korean Academy of Periodontology.)

Background: Oxyresveratrol is the main constituent of mulberries and has many bioactive properties beneficial to human health. The purpose of this study was to assess the anti-inflammatory effects of oxyresveratrol on in vitro periodontitis model., Methods: Human periodontal ligament cells were treated with oxyresveratrol (0, 10, and 20 µg/mL) for 72 h. Cell viability and flow cytometry assays were performed. To investigate anti-inflammatory effect of oxyresveratrol on periodontal inflammation, nitric oxide production under lipopolysaccharide stimulation was assessed. Next, expression of biomarkers associated periodontal inflammation was evaluated. Scratch wound assay was performed to evaluate cell migration/proliferation potential of oxyresveratrol under lipopolysaccharide stimulation., Results: Periodontal ligament cell toxicity was not observed in oxyresveratrol treatment. Oxyresveratrol treatment significantly inhibited nitric oxide production and reduced MMP-2, MMP-9, TNF-α, IL-6, and IL-8 expressions after lipopolysaccharide stimulation. Regarding cell migration/proliferation, open wound area in oxyresveratrol (33.28 ± 6.80%) was the lowest (p < 0.05)., Conclusions: Within the limits of this study, oxyresveratrol inhibited lipopolysaccharide-induced inflammation in periodontal ligament cells and promoted periodontal ligament cell migration/proliferation. These findings suggest that oxyresveratrol could be valuable for the management of periodontal diseases., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Although intraoral scanning is highly reliable, little is known about its accuracy in young children with limited mouth-opening ability., Aim: To determine the accuracy of intraoral scans based on the degree of mouth opening., Design: To simulate mouth opening in children with primary dentition, three groups (n = 5 per group) were allocated by maximum mouth opening of 30, 37 and 40 mm. After the primary dentition model was connected to a dental phantom, intraoral scanning was performed using iTero and TRIOS4. The scanned files were digitally evaluated. Root mean square values were calculated to assess trueness and precision., Results: iTero showed deviations of three-dimensional trueness of 0.067 ± 0.008, 0.063 ± 0.001 and 0.065 ± 0.005 mm, and TRIOS4 of 0.07 ± 0.002, 0.064 ± 0.003 and 0.066 ± 0.002 mm in the 30, 37 and 40 mm groups, respectively. There were no significant differences in either mouth opening (p > .017) or the intraoral scanners (p > .05). The same statistical results were obtained for precision, with the exception of the 30 mm of mouth opening., Conclusions: Within the limits of this study, limited mouth opening hardly influenced the accuracy of intraoral scanning., (© 2024 The Authors. International Journal of Paediatric Dentistry published by BSPD, IAPD and John Wiley & Sons Ltd.)

Bursaphelenchus xylophilus is the pathogen of pine wilt disease, which can devastate the pine forest ecosystem. Usually, plant cells generate reactive oxygen species (ROS) as a defensive substance or signalling molecules to resist the infection of nematodes. However, little is known about how B. xylophilus effectors mediate the plant ROS metabolism. Here, we identified a pioneer B. xylophilus Prx3-interacting effector 1 (BxPIE1) expressed in the dorsal gland cells and the intestine. Silencing of the BxPIE1 gene resulted in reduced nematode reproduction and a delay in disease progression during parasitic stages, with the upregulation of pathogenesis-related (PR) genes PtPR-3 (class Ⅳ chitinase) and PtPR-9 (peroxidase). The protein-protein interaction assays further demonstrated that BxPIE1 interacts with a Pinus thunbergii class III peroxidase (PtPrx3), which produces H 2 O 2 under biotic stress. The expression of BxPIE1 and PtPrx3 was upregulated during the infection stage. Furthermore, BxPIE1 effectively inhibited H 2 O 2 generating from class III peroxidase and ascorbate can recover the virulence of siBxPIE1-treated B. xylophilus by scavenging H 2 O 2 . Taken together, BxPIE1 is an important virulence factor, revealing a novel mechanism utilized by nematodes to suppress plant immunity., (© 2024 John Wiley & Sons Ltd.)

Statement of Problem: The outcome of photopolymerized 3-dimensional (3D) printing is influenced by the methods used for postprinting cleaning, yet information on postprinting cleaning is sparse., Purpose: The purpose of this in vitro study was to assess the cleaning efficiency and surface and mechanical properties of 3D printed resin according to postprinting cleaning methods., Material and Methods: Specimens were fabricated from a 3D model using resin materials (NextDent C&B MFH and DIOnavi-P. MAX) and were tested for postprinting cleaning methods for 5 minutes with isopropyl alcohol, isopropyl alcohol + ultrasonic, ethyl alcohol, ethyl alcohol + ultrasonic, and ultrasonic alone. Postpolymerization was followed for 5 minutes. The cleaning efficiency, microcomputed tomography (µCT), surface roughness, Vickers hardness, and flexural strength of the specimens were evaluated. The 1-way ANOVA test was performed after considering normality. A post hoc analysis with Bonferroni was also performed (α=.008 or.005)., Results: Ultrasonic in addition to cleaning solutions significantly improved the cleaning efficiency in NextDent C&B MFH specimens (P<.005), whereas ultrasonic did not affect the efficiency in DIOnavi-P. MAX specimens. No significant differences were found in surface roughness by postprinting cleaning methods in either NextDent C&B MFH or DIOnavi-P. MAX (P>.005). No significant changes in surface hardness were observed by postprinting cleaning methods (P>.008). In the NextDent C&B MFH, ethyl alcohol + ultrasonic significantly decreased the flexural strength (P<.005). There were no significant differences in the flexural strength in the DIOnavi-P. MAX (P>.005)., Conclusions: Ethyl alcohol was comparable with isopropyl alcohol for use as a postprinting cleaning solution for both NextDent C&B MFH and DIOnavi-P. MAX. The addition of ultrasonic to cleaning solutions should be applied with caution. These findings suggest that different postprinting cleaning methods can be recommended depending on the 3D printed resin materials., (Copyright © 2024 Editorial Council for The Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Background/purpose: Well-Root PT is a novel bioceramic material developed to overcome limitations of conventional calcium silicate cements. The purpose of this study was to assess the biocompatibility and bioactivity of a premixed putty-type cement, Well-Root PT., Materials and Methods: Identical cylindrical samples were prepared from ProRoot MTA, Biodentine, and Well-Root PT. In vitro calcium weight volume and calcium ion release from the materials were evaluated with scanning electron microscopy and energy-dispersive spectroscopy and inductively coupled plasma-optical emission spectroscopy. An in vivo rat direct pulp capping model was implemented with the materials ( n  = 14 per material). The rats were sacrificed at 7 or 28 days. Hematoxylin and eosin and immunohistochemical analyses were performed., Results: In vitro calcium weight volume was 42.83 ± 8.82 % in ProRoot MTA, 47.05 ± 8.83 % in Biodentine, and 29.99 ± 4.94 % in Well-Root PT. Calcium ion releases from Well-Root PT after 7 and 28 days were similar with those from ProRoot MTA, but lower than those from Biodentine ( P  = 0.001 after 7 and 28 days equally). In an in vivo rat model, hematoxylin and eosin analysis showed no significant differences in inflammatory infiltration ( P  = 0.393) and hard tissue formation scores among the materials ( P  = 0.905). Also, both CD68 and DSPP expression showed similar results, with no significant differences among the materials (equally P  = 0.874 for both markers)., Conclusion: Within the limits of this study, Well-Root PT was comparable to ProRoot MTA and Biodentine in terms of biocompatibility and bioactivity., Competing Interests: The authors have no conflicts of interest relevant to this article., (© 2024 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Background: To evaluate the association between the pulmonary vein (PV) entry site morphology after total anomalous pulmonary vein repair (TAPVC) and postoperative pulmonary vein stenosis (PVS)., Methods: Computed tomography (CT) examination was performed to determine the PV entry site morphology. The width of the PV confluence was divided by the width of the left atrium (LA) to obtain the cPV/LA index. The cPV/LA index was compared between patients with and without postoperative PVS., Results: Fifty-one patients who had undergone CT after TAPVC repair were included, with a median cPV/LA index of 0.5 (interquartile range (IQR) = 0.349-0.654). Among them, 27 patients developed postoperative PVS. The median cPV/LA index after primary TAPVC repair was significantly lower in patients with PVS compared to those without PVS (0.367, IQR = 0.308-0.433 vs. 0.657, IQR = 0.571-0.783, P < 0.0001). Additionally, the cPV/LA index after surgical re-intervention for PVS was significantly smaller in patients who developed recurrent stenosis compared to those who remained free-from re-stenosis after surgical relief (0.459, IQR = 0.349-0.556; vs. 0.706, IQR = 0.628-0.810, P = 0.0045)., Conclusion: A small PV confluence width is associated with the development of postoperative PVS and recurrent stenosis after surgical relief of PVS. Our results suggest that adequate bilateral pulmonary vein lateralization during TAPVC surgery is crucial., Competing Interests: Declaration of competing interest All the authors reported no conflicts of interest., (Copyright © 2024 Formosan Medical Association. Published by Elsevier B.V. All rights reserved.)

Objective: In this study, the impact of inhibiting the PI3K/AKT/NF-κB pathway on lung oxidative damage induced by Echinococcus granulosus cyst fluid was investigated., Methods: Twenty-four mice were randomly assigned to four groups. Three months after inoculation with hydatid cyst segments, mice in group A were treated with intraperitoneal and intratracheal saline injections; mice in group B were administered a caudal vein injection of a PI3K inhibitor, followed by cyst fluid sensitization; mice in group C received an AKT inhibitor via caudal vein, followed by cyst fluid sensitization; and mice in group D were subjected to cyst fluid sensitization without any inhibitor treatment. Cellular changes in lung tissues across all groups were evaluated, including pathological section analysis. Analysis of pulmonary tissue and serum from these mice included the assessment of PI3K/AKT/NF-κB pathway proteins, inflammatory factors, and related mRNA levels., Results: Mice in groups B and C exhibited a higher proportion of M2-type macrophages and significantly lower levels of PI3K/AKT/NF-κB pathway proteins, inflammatory factors (interleukin-6 [IL-6]/tumor necrosis factor-α [TNF-α]), and oxidative markers in lung tissues compared to mice in group D ( P < 0.05)., Conclusion: Our results in this study indicate that activation of the PI3K/AKT/NF-κB pathway contributed to an increase in the M1 macrophage phenotype, leading to enhanced secretion of peroxidases and inflammatory factors. This mechanism plays a crucial role in the oxidative and inflammatory lung damage associated with allergic reactions to E. granulosus cyst fluid., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Li, Wang, Zhou, Wang, Tailaiti, Lin, Bayina, Cao and Ye.)

Background: Cell wall integrity (CWI) is crucial for fungal growth, pathogenesis, and adaptation to extracellular environments. Calcofluor white (CFW) is a cell wall perturbant that inhibits fungal growth, yet little is known about how phytopathogenic fungi respond to the CFW-induced stress., Results: In this study, we unveiled a significant discovery that CFW triggered the translocation of the transcription factor CgCrzA from the cytoplasm to the nucleus in Colletotrichum gloeosporioides. This translocation was regulated by an interacting protein, CgMkk1, a mitogen-activated protein kinase involved in the CWI pathway. Further analysis revealed that CgMkk1 facilitated nuclear translocation by phosphorylating CgCrzA at the Ser280 residue. Using chromatin immunoprecipitation sequencing, we identified two downstream targets of CgCrzA, namely CgCHS5 and CgCHS6, which are critical for growth, cell wall integrity, and pathogenicity as chitin synthase genes., Conclusions: These findings provide a novel insight into the regulatory mechanism of CgMkk1-CgCrzA-CgChs5/6, which enables response of the cell wall inhibitor CFW and facilitates infectious growth for C. gloeosporioides., (© 2024. The Author(s).)

Pinus is an important economic tree species, but pine wilt disease (PWD) seriously threatens the survival of pine trees. PWD caused by Bursaphelenchus xylophilus is a major quarantine disease worldwide that causes significant economic losses. However, more information about its molecular pathogenesis is needed, resulting in a lack of effective prevention and treatment measures. In recent years, effectors have become a hot topic in exploring the molecular pathogenic mechanism of pathogens. Here, we identified a specific effector, BxNMP1, from B. xylophilus . In situ hybridization experiments revealed that BxNMP1 was specifically expressed in dorsal gland cells and intestinal cells, and RT-qPCR experiments revealed that BxNMP1 was upregulated in the early stage of infection. The sequence of BxNMP1 was different in the avirulent strain, and when BxNMP1 -silenced B. xylophilus was inoculated into P. thunbergii seedlings, the disease severity significantly decreased. We demonstrated that BxNMP1 interacted with the thaumatin-like protein PtTLP-L2 in P. thunbergii . Additionally, we found that the β-1,3-glucanase PtGLU interacted with PtTLP-L2. Therefore, we hypothesized that BxNMP1 might indirectly interact with PtGLU through PtTLP-L2 as an intermediate mediator. Both targets can respond to infection, and PtTLP-L2 can enhance the resistance of pine trees. Moreover, we detected increased salicylic acid contents in P. thunbergii seedlings inoculated with B. xylophilus when BxNMP1 was silenced or when the PtTLP-L2 recombinant protein was added. In summary, we identified a key virulence effector of PWNs, BxNMP1. It positively regulates the pathogenicity of B. xylophilus and interacts directly with PtTLP-L2 and indirectly with PtGLU. It also inhibits the expression of two targets and the host salicylic acid pathway. This study provides theoretical guidance and a practical basis for controlling PWD and breeding for disease resistance.

Competing Interests: Declaration of competing interest All authors declare that they have no conflicts of interest.

Background/aim: Frondoside A is a sea cucumber extract which is well known for its anti-inflammatory and immunomodulatory properties. The purpose of this study was to evaluate the effect of Frondoside A application in the alveolar socket on inflammatory responses after delayed replantation in rat teeth., Materials and Methods: Human periodontal ligament cells were cultured and exposed to Frondoside A. Cell-counting kit-8 assay was performed to evaluate the cell viability and nitric oxide assay was performed to assess the anti-inflammatory effect of Frondoside A. Molars were extracted from 32 Sprague-Dawley rats and randomly divided into control and Frondoside A groups. After 30 min of extra-oral dry time, molars were replanted. In the Frondoside A group, Frondoside A solution was applied in the alveolar socket before replantation. The animals were sacrificed after 28 days and histologically and immunohistochemically evaluated., Results: 0.5 μM Frondoside A showed higher cellular viability at 6 h and lower production of nitric oxide compared with other Frondoside A solutions (p < .05). The Frondoside A group demonstrated lower inflammatory resorption scores in both middle 1/3 and apical 1/3 of root compared to the control group (p < .05). The Frondoside A group showed lower levels of expression in both cathepsin K and CD45 compared with the control group (p < .05)., Conclusions: Within the limits of this study, intra-alveolar delivery of Frondoside A alleviates inflammatory root resorption in delayed replantation of rat teeth., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Sufficient alveolar housing is fundamental to ensure the favorable prognosis of autotransplanted teeth. However, little is known about the alveolar ridge profiles of the anterior maxilla in children. Therefore, this study aimed to investigate the alveolar ridge profiles of the anterior maxilla in growing children., Methods: A total of 169 Korean children aged 10-13 years old were included in this retrospective study. Demographic data and cone-beam computed tomography scans of 244 maxillary central incisors were collected. For each tooth, the alveolar ridge thickness was determined at 1-, 3-, and 5-mm levels below the cemento-enamel junction. The alveolar process height and labial concavity were also evaluated. Statistical analyses were performed to evaluate differences according to age and sex., Results: The alveolar ridge thickness did not differ according to age. The alveolar ridge thickness in males was significantly greater than that in females among those aged 10 and 11 years. The average alveolar process height was 17.43 ± 2.22 mm. The alveolar process height hardly changed with age from 10-13 years, whereas the alveolar process height of males was significantly greater compared with that of females among those aged 10 years. The labial concavity was approximately 150˚, with no significant differences between age and sex., Conclusions: Within the limits of this study, the anatomy of the alveolar ridge in the maxillary central incisors showed sufficient capacity as a recipient site for autotransplantation using mandibular premolars, especially in males. These findings serve as a valuable resource for facilitating autotransplantation in children., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Context: Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, has been shown to exhibit anti-depressive effects in clinical trials. However, the direct mechanism underlying its effect on neuroinflammation remains unclear. Neuroinflammatory reaction from astrocytes leads to depression, and our previous study found that gap junction disorder between astrocytes aggravated neuroinflammatory reaction in depressed mice., Objective: To investigate the potential mechanism of celecoxib's effects on astrocytic gap junctions during the central nervous inflammation-induced depression., Materials & Methods: Stereotaxic injection of lipopolysaccharide (LPS) into the prefrontal cortex (PFC) to establish a model of major depressive disorder (MDD). Celecoxib was administrated into PFC 15 min after LPS injection. The depressive performance was tested by tail suspension test and forced swimming test, and the levels of proinflammation cytokines were determined at mRNA and protein levels. Resting-state functional connection (rsFC) was employed to assess changes in the default mode network (DMN). Additionally, astrocytic gap junctions were also determined by lucifer yellow (LY) diffusion and transmission electron microscope (TEM), and the expression of connexin 43 (Cx43) was measured by western blotting, quantitative polymerase chain reaction, and immunofluorescence., Results: LPS injection induced significant depressive performance, which was ameliorated by celecoxib treatment. Celecoxib also improved rsFC in the DMN. Furthermore, celecoxib improved astrocytic gap junctions as evidenced by increased LY diffusion, shortened gap junction width, and normalized levels of phosphorylated Cx43. Celecoxib also blocked the phosphorylation of p65, and inhibition of p65 abolished the improvement of Cx43., Discussion & Conclusion: Anti-depressive effects of celecoxib are mediated, at least in part, by the inhibition of nuclear factor- kappa B (NF-κB) and the subsequent improvement of astrocytic gap junction function., Competing Interests: Declaration of Competing Interest The authors declare that there are no competing interests to declare., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: This study aimed to assess the biocompatibility and bioactivity of a dual-cured resin-based calcium silicate cement in vitro and in vivo., Methods: For in vitro analyses, standardized samples were prepared using TheraCal LC, TheraCal PT, and ProRoot MTA. The amount of residual monomer released from TheraCal LC and TheraCal PT was assessed using liquid chromatography/mass spectrometry. Calcium ion release from the materials was evaluated using inductively coupled plasma-optical emission spectroscopy. Scanning electron microscopy and energy-dispersive X-ray spectroscopy were used to determine the calcium weight volume in the materials. For in vivo analysis, a rat direct pulp capping model with TheraCal LC, TheraCal PT, and ProRoot MTA groups (n = 16 per group) was used. The rats were euthanized after 7 or 28 days, and histological and immunohistochemical analyses (CD68 and DSPP) were performed., Results: Bisphenol A-glycidyl methacrylate and polyethylene glycol dimethacrylate release from TheraCal PT was lower than that from TheraCal LC (P < .05). Similar results were obtained for calcium-ion release and calcium weight volume, with ProRoot MTA showing the highest values. In the in vivo evaluation, TheraCal PT showed significantly greater hard tissue formation than TheraCal LC (P < .017). TheraCal PT showed lower CD68 expression and greater DSPP expression than TheraCal LC (P < .017). There were no significant differences in the expression of CD68 or DSPP between the TheraCal PT and ProRoot MTA groups., Conclusions: Within the limitations of this study, the biocompatibility and bioactivity of TheraCal PT could be comparable to those of ProRoot MTA., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

A 21-year-old woman with severe pulmonary hypertension and circulatory collapse was referred to our hospital for possible lung transplantation with extracorporeal membrane oxygenation support. Computed tomography revealed severe stenosis of all 4 pulmonary veins, and fibrosing mediastinitis was suspected. Surgical reconstruction of the pulmonary veins was performed, and extracorporeal membrane oxygenation support was weaned off. After surgery, pulmonary vascular resistance normalized. This successful case demonstrates that surgical pulmonary venous reconstruction is an important treatment for fibrosing mediastinitis induced by pulmonary venous stenosis and pulmonary hypertension., (Copyright © 2023 International Society for the Heart and Lung Transplantation. Published by Elsevier Inc. All rights reserved.)

Purpose: The objective of this study was to investigate the influence of photodynamic therapy (PDT) employing different, lower 5-aminolevulinic acid (ALA) dosages on the proliferative activity of Cutibacterium acnes (C. acnes)., Methods: In this in vitro bacterial experiment, we examined the effects of PDT using different doses of ALA (0.05 mmol/L; 0.1 mmol/L; 0.5 mmol/L; 1.0 mmol/L; 2.5 mmol/L). To elucidate the underlying mechanisms, we assessed colony-forming units (CFUs), bacterial staining for live/dead, antioxidant enzyme activity, and gene expression of oxidative stress markers following treatment with different doses of ALA-PDT., Results: Our findings demonstrate that CFU, bacterial staining for live/dead, as well as the activity and gene expression of superoxide dismutase (SOD) and catalase (CAT), all exhibited significant increases when the ALA concentration was 0.1/0.5 mmol/L. However, both CFU and cell growth of C. acnes decreased when the ALA concentration reached 1.0 mmol/L., Conclusion: Lower concentration of ALA-PDT (0.1/0.5 mmol/L) appears to promote the growth of C.acnes while higher doses (1.0 /2.5 mmol/L) are associated with eradication. The procedure is possibly mediated by the activation of antioxidant-related genes and enzyme expression in cells., Competing Interests: Declaration of Competing Interest The authors have declared that no competing interest exists., (Copyright © 2023. Published by Elsevier B.V.)

Background and Objectives : In spite of the oral environment being healing-prone, its dynamic changes may affect wound healing. The purpose of this study was to assess the oral wound healing effect of Angelica gigas Nakai (AG) prepared by hot-melt extrusion. Materials and Methods : Human gingival fibroblast (HGF) cells were treated with AG or AG via hot-melt extrusion (AGH) for 24 h to determine the optimal concentration. For evaluating the anti-inflammatory effect of AG and AGH, a nitric oxide assay was performed under lipopolysaccharide (LPS) stimulation. The wound-healing effects of AG and AGH were evaluated using cell proliferation/migration assays and wound-healing marker expression through qRT-PCR. Results : Both AG and AGH showed no cytotoxicity on HGH cells. Regarding nitric oxide production, AGH significantly decreased LPS-induced nitric oxide production ( p < 0.05). AGH showed a significantly positive result in the cell proliferation/cell migration assay compared with that in AG and the control. Regarding wound healing marker expression, AGH showed significantly greater VEGF and COL1α1 expression levels than those in the others ( p < 0.05), whereas α-SMA expression was significantly different among the groups. Conclusions : Within the limits of this study, AGH accelerated oral wound healing in vitro.

Background: To explore the value of early oral nutritional supplements (ONS) in patients with nasopharyngeal carcinoma (NPC) treated with concurrent chemoradiotherapy (CCRT)., Methods: Patients with newly diagnosed II-IVA stage NPC were analyzed and divided into Early and Routine ONS groups according to whether they received early ONS at the beginning of CCRT. Changes in nutritional indicators, incidence of treatment-related toxicity, radiation interruption, and completion of CCRT were compared., Results: In total, 161 patients with NPC were analyzed, including 72 in the Early ONS group and 89 in the Routine ONS group. Multivariate analysis showed that early ONS was an independent protective factor for concurrent chemotherapy ≥2 cycles, and a protective factor against ≥grade 3 radiation-induced oral mucositis (RIOM) and weight loss >5%. In stage III-IVA patients, early ONS was beneficial in decreasing the risk of severe malnutrition., Conclusions: Early ONS can improve nutritional outcomes, reduce RIOM, and enhance treatment adherence., (© 2023 Wiley Periodicals LLC.)

Visceral pain (VP) is caused by internal organ disease. VP is involved in nerve conduction and related signaling molecules, but its specific pathogenesis has not yet been fully elucidated. Currently, there are no effective methods for treating VP. The role of P2X2/3 in VP has progressed. After visceral organs are subjected to noxious stimulation, cells release ATP, activate P2X2/3, enhance the sensitivity of peripheral receptors and the plasticity of neurons, enhance sensory information transmission, sensitize the central nervous system, and play an important role in the development of VP. However, antagonists possess the pharmacological effect of relieving pain. Therefore, in this review, we summarize the biological functions of P2X2/3 and discuss the intrinsic link between P2X2/3 and VP. Moreover, we focus on the pharmacological effects of P2X2/3 antagonists on VP therapy and provide a theoretical basis for its targeted therapy., Competing Interests: Declaration of Competing Interest There is no conflict of interest in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Parkinson's disease (PD) is pathologically manifested by the aggregation of α-synuclein, which has been envisioned as a promising disease-modifying target for PD. Here, we identified 20C, a bibenzyl compound derived from Gastrodia elata, able to inhibit the aggregation of A53T variants of α-synuclein directly in vitro. Computational analysis revealed that 20C binds to cavities in mature α-synuclein fibrils, and it indeed displays a strong interaction with α-synuclein and reduced their β-sheet structure by microscale thermophoresis and circular dichroism, respectively. Moreover, incubating neural cells with 20C reduced the amounts of α-synuclein inclusions significantly. The treatment of A53T α-Syn transgenic mice with 20C significantly reduces the toxic α-synuclein levels, improves behavioral performance, rescues dopaminergic neuron, and enhances functional connections between SNc and PD associated brain areas. The transcriptome analysis of SNc demonstrated that 20C improves mitochondrial dynamics, which protects mitochondrial morphology and function against α-synuclein induced degeneration. Overall, 20C appears to be a promising candidate for the treatment of PD., (© 2023. The Author(s).)

Lipase is involved in lipid hydrolysis, which is related to nematodes' energy reserves and stress resistance. However, the role of lipases in Bursaphelenchus xylophilus, a notorious plant-parasitic nematode responsible for severe damage to pine forest ecosystems, remains largely obscure. Here, we characterized a class III lipase as a candidate effector and named it BxLip-3. It was transcriptionally up-regulated in the parasitic stages of B. xylophilus and specifically expressed in the oesophageal gland cells and the intestine. In addition, BxLip-3 suppressed cell death triggered by the pathogen-associated molecular patterns PsXEG1 and BxCDP1 in Nicotiana benthamiana, and its Lipase-3 domain is essential for immunosuppression. Silencing of the BxLip-3 gene resulted in a delay in disease onset and increased the activity of antioxidant enzymes and the expression of pathogenesis-related (PR) genes. Plant chitinases are thought to be PR proteins involved in the defence system against pathogen attack. Using yeast two-hybrid and co-immunoprecipitation assays, we identified two class I chitinases in Pinus thunbergii, PtChia1-3 and PtChia1-4, as targets of BxLip-3. The expression of these two chitinases was up-regulated during B. xylophilus inoculation and inhibited by BxLip-3. Overall, this study illustrated that BxLip-3 is a crucial virulence factor that plays a critical role in the interaction between B. xylophilus and host pine., (© 2023 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Objective: To determine the pathogenesis and molecular targets of anaphylaxis caused by hydatid cyst fluid leakage., Methods: First, Balb/c mice were infected with Echinococcus granulosus, and then the anaphylaxis model was developed. The mice were separated into: anaphylaxis caused by the cystic echinococcosis group (ANPC), the cystic echinococcosis without anaphylaxis group (CE group), and the normal control group (CTRL). Following this, the spleen tissue was collected for microRNA (miRNA) sequencing. Using bioinformatics analysis, differentially expressed miRNAs (DEMs) were identified. Then, through the use of protein-protein interaction (PPI) networks, the key target genes for miRNA regulation associated with echinococcosis-induced anaphylaxis were identified., Results: ANPC and CE groups have 29 and 39 DEMs compared to the CTRL group, respectively. Based on these 25 DEMs, interactions between miRNA and mRNA were screened, and 174 potential target genes were identified. We performed gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis on these 174 target genes, and the results revealed that the three pathways with the highest enrichment were the PI3K-Akt signaling pathway, FoxO signaling pathway, and Focal adhesion. The interaction analysis of PPI and miRNA-hub gene networks revealed that interleukin 6 (IL-6) was regulated by miR-146a-5p and miR-149-5p, while IL-10 was regulated by miR-29b-3p and miR-29c-3. Using reverse transcription polymerase chain reaction, we found that the miRNAs regulating IL-6 and IL-10 were significantly upregulated in the ANPC group, and there are three pathways involved in that process: Pathways of PI3K-Akt signaling, FoxO signaling, and Focal adhesion. IL-6 and IL-10 play an important role in cellular pyroptosis and apoptosis. Therefore, the aforementioned results provide significant reference value for elucidating the mechanism of cellular pyroptosis and apoptosis in echinococcosis-induced anaphylaxis, and for formulating tissue and organ protection strategies for patients with cystic echinococcosis when anaphylaxis is triggered by hydatid cyst rupture., (© 2023 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.)

Objective: To investigate the changes in memory T cells and the related factors in mice by the establishment of a BALB/c mouse model of Echinococcus granulosus-induced sensitization., Methods: A sensitized BALB/c mouse model was established by intraperitoneal injection of E. granulosus. A control group (CTRL), a nonsensitized group infected with E. granulosus (CE), and a sensitized group infected with E. granulosus (ANPC) were set up. The pathological changes in lung tissue in mice, the change in memory T cells (CD4 Tm), and the change in peripheral blood nucleated interleukin-23 (IL-23) were detected using HE staining, flow cytometry, and liquid-phase multiple protein quantification techniques, respectively., Results: The individual percentage of mouse memory T cells was 9.14 ± 0.45, 25.23 ± 0.17, and 13.29 ± 0.32 in the CTRL, CE, and ANPC groups, respectively. The percentage of memory T cells in the ANPC group was higher than that in the CTRL group (t = 18.410, p < .001) but lower than that in the CE group (t = -80.147, p < .001). The levels of IL-23 in peripheral blood of mice in the CTRL, CE, and ANPC groups were 225.76 ± 27.16, 359.21 ± 28.67, and 215.69 ± 22.69, respectively. The level of IL-23 in peripheral blood of mice in the ANPC group was lower than that in the CE group (t = 9.609, p < .001), and there was no statistical difference with the CTRL group (t = 0.697, p = .502)., Conclusion: In the BALB/c mouse model of E. granulosus-induced sensitization, the expression of IL-23 in peripheral blood increased, and the memory T cell proliferated and became activated; there was a decrease in the content of IL-23 in peripheral blood and number of activated memory T cells in the sensitization group infected with E. granulosus. The E. granulosus-induced allergic reaction was related to IL-23 and the activation of memory T cells., (© 2023 The Authors. Immunity, Inflammation and Disease published by John Wiley & Sons Ltd.)

Selecting suitable promoters to drive gene overexpression can provide significant insight into the development of engineered bacteria. In this study, we analyzed the transcriptome data of Burkholderia pyrrocinia JK-SH007 and identified 54 highly expressed genes. The promoter sequences were located using genome-wide data and scored using the prokaryotic promoter prediction software BPROM to further screen out 18 promoter sequences. We also developed a promoter trap system based on two reporter proteins adapted for promoter optimization in B. pyrrocinia JK-SH007: firefly luciferase encoded by the luciferase gene set ( Luc ) and trimethoprim ( TP )-resistant dihydrofolate reductase ( TP r ). Ultimately, eight constitutive promoters were successfully inserted into the probe vector and transformed into B. pyrrocinia JK-SH007. The transformants were successfully grown on Tp antibiotic plates, and firefly luciferase expression was determined by measuring the relative light unit (RLU). Five of the promoters (P4, P9, P10, P14, and P19) showed 1.01-2.51-fold higher activity than the control promoter λ phage transcriptional promoter (PRPL). The promoter activity was further validated via qPCR analysis, indicating that promoters P14 and P19 showed stable high transcription levels at all time points. Then, GFP and RFP proteins were overexpressed in JK-SH007. In addition, promoters P14 and P19 were successfully used to drive gene expression in Burkholderia multivorans WS-FJ9 and Escherichia coli S17-1. The two constitutive promoters can be used not only in B. pyrrocinia JK-SH007 itself to gene overexpression but also to expand the scope of application.

Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus ), is a major quarantine forest disease that poses a threat to various pine species, including Pinus massoniana (masson pine), worldwide. Breeding of PWN-resistant pine trees is an important approach to prevent the disease. To expedite the production of PWN-resistant P. massoniana accessions, we investigated the effects of maturation medium treatments on somatic embryo development, germination, survival, and rooting. Furthermore, we evaluated the mycorrhization and nematode resistance of regenerated plantlets. Abscisic acid was identified as the main factor affecting maturation, germination, and rooting of somatic embryos in P. massoniana , resulting in a maximum of 34.9 ± 9.4 somatic embryos per ml, 87.3 ± 9.1% germination rate, and 55.2 ± 29.3% rooting rate. Polyethylene glycol was identified as the main factor affecting the survival rate of somatic embryo plantlets, with a survival rate of up to 59.6 ± 6.8%, followed by abscisic acid. Ectomycorrhizal fungi inoculation with Pisolithus orientalis enhanced the shoot height of plantlets regenerated from embryogenic cell line (ECL) 20-1-7. Ectomycorrhizal fungi inoculation also improved the survival rate of plantlets during the acclimatization stage, with 85% of mycorrhized plantlets surviving four months after acclimatization in the greenhouse, compared with 37% non-mycorrhized plantlets. Following PWN inoculation, the wilting rate and the number of nematodes recovered from ECL 20-1-7 were lower than those recovered from ECL 20-1-4 and 20-1-16. The wilting ratios of mycorrhizal plantlets from all cell lines were significantly lower than those of non-mycorrhizal regenerated plantlets. This plantlet regeneration system and mycorrhization method could be used in the large-scale production of nematode-resistance plantlets and to study the interaction between nematode, pines, and mycorrhizal fungi., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Chen, Fei, Xia, Ke, Ye and Zhu.)

Ischemic stroke is characterized by the presence of reactive microglia. However, its precise involvement in stroke etiology is still unknown. We used metabolic profiling and showed that chemokine like factor 1 (CKLF1) causes acute microglial inflammation and metabolic reprogramming from oxidative phosphorylation to glycolysis, which was reliant on the AMP-activated protein kinase (AMPK)-mammalian target of rapamycin (mTOR)-hypoxia inducible factor 1α (HIF-1α) signaling pathway. Once activated, microglia enter a chronic tolerant state as a result of widespread energy metabolism abnormalities, which reduces immunological responses, including cytokine release and phagocytosis. Metabolically dysfunctional microglia were also found in mice using genome-wide RNA sequencing after chronic administration of CKLF1, and there was a decrease in the inflammatory response. Finally, we showed that the loss of CKLF1 reversed the defective immune response of microglia, as indicated by the maintenance its phagocytosis to neutrophils, thereby mitigating the long-term outcomes of ischemic stroke. Overall, CKLF1 plays a crucial role in the relationship between microglial metabolic status and immune function in stroke, which prepares a potential therapeutic strategy for ischemic stroke., (© 2023. The Author(s).)

Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Anthracnose is one of the most widespread and destructive diseases on Chinese fir. Colletotrichum cangyuanense, Colletotrichum fructicola, Colletotrichum gloeosporioides, and Colletotrichum siamense are the causal agents of anthracnose on Chinese fir. A rapid and accurate diagnosis of different pathogens is critical for the disease management., Results: Phylogenetic tree and sequence similarity analysis showed that the single-locus ApMat provides superior phylogenetic information and is an appropriate target to distinguish C. cangyuanense, C. fructicola, C. gloeosporioides, and C. siamense. The real-time PCR assays with the primer sets of MQ-F/R, 1#C-F/R, YK-F/R, and WZ-F/R, and corresponding TaqMan probes of MQ-P, 1#C-P, YK-P, and WZ-P were specific for C. cangyuanense, C. fructicola, C. gloeosporioides, and C. siamense, respectively. The sensitivity tests showed that the lowest amount of gDNA that the PCRs can detect was 1 ng of genomic DNA. The validity of the assays was confirmed by directly detecting the pathogens from both the fungal culture and infected Chinese fir., Conclusion: These results demonstrated the potential of the TaqMan real-time PCR targeting the ApMat gene to provide rapid, specific, and reliable molecular detection of C. fructicola, C. gloeosporioides, C. siamense, and C. cangyuanense, respectively. The data also provided a reference solution for the identification of species within Colletotrichum gloeosporioides species complex (CGSC), which share similar morphological characteristics. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Pine wilt disease, caused by Bursaphelenchus xylophilus , results in tremendous economic loss in conifer production every year. To disturb the host immune responses, plant pathogens secrete a mass of effector proteins that facilitate the infection process. Although several effectors of B. xylophilus have been identified, detailed mechanisms of their functions remain largely unexplored. Here, we reveal two novel B. xylophilus Kunitz effectors, named BxKU1 and BxKU2, using different infection strategies to suppress immunity in Pinus thunbergii . We found that both BxKU1 and BxKU2 could suppress PsXEG1-triggered cell death and were present in the nucleus and cytoplasm in Nicotiana benthamiana . However, they had different three-dimensional structures and various expression patterns in B. xylophilus infection. In situ hybridization experiments showed that BxKU2 was expressed in the esophageal glands and ovaries, whereas BxKU1 was only expressed in the esophageal glands of females. We further confirmed that the morbidity was significantly decreased in P. thunbergii infected with B. xylophilus when BxKU1 and BxKU2 were silenced. The silenced BxKU2I, but not BxKU1 , affected the reproduction and feeding rate of B. xylophilus . Moreover, BxKU1 and BxKU2 targeted to different proteins in P. thunbergii , but they all interacted with thaumatin-like protein 4 (TLP4) according to yeast two-hybrid screening. Collectively, our study showed that B. xylophilus could incorporate two Kunitz effectors in a multilayer strategy to counter immune response in P. thunbergii , which could help us better understand the interaction between plant and B. xylophilus .

The pinewood nematode, Bursaphelenchus xylophilus , has been determined as one of the world's top ten plant-parasitic nematodes. It causes pine wilt, a progressive disease that affects the economy and ecologically sustainable development in East Asia. B. xylophilus secretes pathogenic proteins into host plant tissues to promote infection. However, little is known about the interaction between B. xylophilus and pines. Previous studies reported transthyretin proteins in some species and their strong correlation with immune evasion, which has also been poorly studied in B. xylophilus . In this study, we cloned and functionally validated the B. xylophilus pathogenic protein BxTTR-52, containing a transthyretin domain. An in situ hybridization assay demonstrated that BxTTR-52 was expressed mainly in the esophageal glands of B. xylophilus . Confocal microscopy revealed that BxTTR-52-RFP localized to the nucleus, cytoplasm, and plasma membrane. BxTTR-52 recombinant proteins produced by Escherichia coli could be suppressed by hydrogen peroxide and antioxidant enzymes in pines. Moreover, silencing BxTTR-52 significantly attenuated the morbidity of Pinus thunbergii infected with B. xylophilus . It also suppressed the expression of pathogenesis-related genes in P. thunbergii . These results suggest that BxTTR-52 suppresses the plant immune response in the host pines and might contribute to the pathogenicity of B. xylophilus in the early infection stages.

Plant growth-promoting rhizobacteria (PGPR) are important members of soil microbial communities. In this study, the effects of several PGPR on the growth of Carya illinoinensis plants, the microbial community composition and soil nutrients were investigated by inoculation tests to identify excellent PGPR strains. The experiment showed that after PGPR application, the plant height, ground diameter, and dry weight of C. illinoinensis were significantly increased compared with those of the control group, and Bacillus velezensis YH20 had the most significant effect in promoting growth (p < 0.05). In addition, all the PGPRs used for inoculation promoted plant root growth, and the Brevibacillus reuszeri MPT17 strain had the most significant promoting effect on plant root growth (p < 0.05). The application of PGPRs also affected the nutrient levels in plants and plant rhizosphere soil. For example, compared with the control, the levels of available phosphorus and potassium in rhizosphere soil and the total potassium content in plant roots were significantly increased under Br. reuszeri MPT17 treatment (p < 0.05). The experiment showed that the relative abundance of Mortierella, Dictyophora, and Bacillus in the rhizosphere soil increased significantly after the application of PGPR (p < 0.05). These genera could effectively improve the rate of soil nutrient use, antagonize plant pathogenic bacteria, and promote plant growth. This study provides basic reference data regarding the use of PGPR to improve the microecological environment and promote the growth and development of C. illinoinensis plants., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The migratory plant-parasitic nematode Bursaphelenchus xylophilus is the pathogen of the pine wilt disease (PWD), causing serious damage to pine forests in China. During the process of plant resistance to multiple pathogens, plant immunity plays a key role. In this current study, the pathogen-associated molecular pattern (PAMP) BxCDP1 in B. xylophilus has been identified, but the host target protein of BxCDP1 and its key amino acid region inducing the plant immunity have yet to be elucidated. We found that BxCDP1 could trigger superoxide production, H 2 O 2 production, and callose deposits. A RING-H2 finger protein 1 (RHF1) of Pinus thunbergii was screened and characterized as a target protein of BxCDP1 by yeast two-hybrid and co-immunoprecipitation (Co-IP). Moreover, two peptides (namely M9 and M16) proved to be key regions of BxCDP1 to induce PAMP-triggered immunity (PTI) in Nicotiana benthamiana , which also induced the expression of pathogenesis-related (PR) genes ( PtPR-3 , PtPR-4 , and PtPR-5 ) in P. thunbergii and enhanced the resistance of the host to B. xylophilus . These results indicate that BxCDP1 plays a critical role in the interaction between B. xylophilus and P. thunbergii , and both peptides M9 and M16 have the potential to be developed and utilized as immune inducers of pines against B. xylophilus in future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hu, Wu, Wen, Ye, Qiu, Rui and Zhang.)

Bursaphelenchus xylophilus is the most economically important species of migratory plant-parasitic nematodes (PPNs) and causes severe damage to forestry in China. The successful infection of B. xylophilus relies on the secretion of a repertoire of effector proteins. The effectors, which suppress the host pine immune response, are key to the facilitation of B. xylophilus parasitism. An exhaustive list of candidate effectors of B. xylophilus was predicted, but not all have been identified and characterized. Here, an effector, named BxSCD3, has been implicated in the suppression of host immunity. BxSCD3 could suppress pathogen-associated molecular patterns (PAMPs) PsXEG1- and INF1-triggered cell death when it was secreted into the intracellular space in Nicotiana benthamiana. BxSCD3 was highly up-regulated in the early infection stages of B. xylophilus . BxSCD3 does not affect B. xylophilus reproduction, either at the mycophagous stage or the phytophagous stage, but it contributes to the virulence of B. xylophilus. Moreover, BxSCD3 significantly influenced the relative expression levels of defense-related (PR) genes PtPR-3 and PtPR-6 in P inus thunbergii in the early infection stage. These results suggest that BxSCD3 is an important toxic factor and plays a key role in the interaction between B. xylophilus and host pine.

The high incidence of treatment-resistant pain calls for the urgent preclinical translation of new analgesics. Understanding the behavioral readout of pain in animals is crucial for efficacy evaluation when developing novel analgesics. Mas-related G protein-coupled receptor D-positive (Mrgprd + ) and transient receptor potential vanilloid 1-positive (TRPV1 + ) sensory neurons are two major non-overlapping subpopulations of C-fiber nociceptors. Their activation has been reported to provoke diverse nocifensive behaviors. However, what kind of behavior reliably represents subjectively conscious pain perception needs to be revisited. Here, we generated transgenic mice in which Mrgprd + or TRPV1 + sensory neurons specifically express channelrhodopsin-2 (ChR2). Under physiological conditions, optogenetic activation of hindpaw Mrgprd + afferents evoked reflexive behaviors (lifting, etc.), but failed to produce aversion. In contrast, TRPV1 + afferents activation evoked marked reflexive behaviors and affective responses (licking, etc.), as well as robust aversion. Under neuropathic pain conditions induced by spared nerve injury (SNI), affective behaviors and avoidance can be elicited by Mrgprd + afferents excitation. Mechanistically, spinal cord-lateral parabrachial nucleus (lPBN) projecting neurons in superficial layers (lamina I-II o ) were activated by TRPV1 + nociceptors in naïve conditions or by Mrgprd + nociceptors after SNI, whereas only deep spinal cord neurons were activated by Mrgprd + nociceptors in naïve conditions. Moreover, the excitatory inputs from Mrgprd + afferents to neurons within inner lamina II (II i ) are partially gated under normal conditions. Altogether, we conclude that optogenetic activation of the adult Mrgprd + nociceptors drives non-pain-like reflexive behaviors via the deep spinal cord pathway under physiological conditions and drives pain-like affective behaviors via superficial spinal cord pathway under pathological conditions. The distinct spinal pathway transmitting different forms of nocifensive behaviors provides different therapeutic targets. Moreover, this study appeals to the rational evaluation of preclinical analgesic efficacy by using comprehensive and suitable behavioral assays, as well as by assessing neural activity in the two distinct pathways., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Su, Wu, Xu, Wang, Chen, Ye, Maimaitiabula, Liu, Sun and Zhang.)

Background: The pine wilt disease (PWD) caused by Bursaphelenchus xylophilus is a devastating forest disease and its pathogenesis remains unclear. Secreted enzymes and proteins are important pathogenicity determinants and Bx-FAR-1 is an important pathogenic protein involved in the interaction between pine and B. xylophilus. However, the function of the Bx-FAR-1 protein in monitoring and prevention PWD remains unknown., Results: We found a small peptide of B. xylophilus effector Bx-FAR-1 is sufficient for immunosuppression function in Nicotiana benthamiana. Transient expression of Bx-FAR-1 in N. benthamiana revealed that nuclear localization is required for its function. The results of the ligand binding test showed that Bx-FAR-1 protein had the ability to bind fatty acid and retinol. We demonstrated that Bx-FAR-1 targeted to the nuclei of Pinus thunbergii using the polyclonal antibody by immunologic approach. The content of jasmonic acid (JA) was significantly increased in P. thunbergii infected with B. xylophilus when Bx-FAR-1 was silenced. We identified an F-box protein as the host target of Bx-FAR-1 by yeast two-hybrid and co-immunoprecipitation. Moreover, we found that Pt-F-box-1 was up-regulated during B. xylophilus infection and the expression of Pt-F-box-1 was increased in Bx-FAR-1 double-stranded RNA (dsRNA)-treated host pines., Conclusion: This study illustrated that Bx-FAR-1 might mediate the JA pathway to destroy the immune system of P. thunbergii, indicating that PWN likely secretes effectors to facilitate parasitism and promote infection, which could better reveal the pathogenesis mechanisms of B. xylophilus and would be beneficial for developing disease control strategies., (© 2022 Society of Chemical Industry.)

Pinus thunbergii Parl. (Japanese black pine), an evergreen species, is distributed along the seacoasts of China. In addition, this species has been planted along seacoasts as a windbreak to prevent soil erosion due to its resistance to salt and various environmental stresses. It can also be found in public parks and gardens due to its exquisite appearance and toughness. In August 2020, needle blight symptoms were found on several black pine trees in Sichuan Province, China. Further surveys showed that these symptoms are common. The disease incidence is less than 30% while severity of the disease is high. The tips of old needles first turn grayish green that developed into brown bands ranging from 1 to 2 mm. To determine the pathogen, small needle pieces (3-4 mm2 long) from the margin of fresh lesions were surface-sterilized for 30 s in 75% ethanol, follow by 1% NaOCl for 90 s, then washed three times with sterile water, and then were placed on potato dextrose agar (PDA) with 0.1 mg/mL ampicillin and incubated at 25°C. Pure cultures of 8 isolates were obtained by monosporic isolation, and a representative isolate (SC03) was deposited in the Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University. When cultured on Spezieller Nährstoffarmer Agar medium (Leslie and Summerell, 2006), the SC03 colony was beige-white, cottony from top view and pale orange near the center on the reverse side. The daily growth rate was 11.8 mm/day at 25°C in the dark. Microscopic observations showed hyaline and septate hyphae, slightly curved macroconidia with two to three septa measuring 17.5 - 30 × 3.7 - 7.5 μm (23.2 × 5.7 on average), and aseptate microconidia measuring 7.5 - 12.5 × 2.5 - 5.0 μm, (9.7 × 4.3 on average). The morphological characteristics of conidia and other structures of SC03 matched those of the Fusarium fujikuroi species complex (FFSC) (Abdalla et al. 2000). For accurate identification, translation elongation factor 1-alpha（TEF-1α） , and the second largest RNA polymerase subunit (RPB2) were amplified and sequenced using the primer pairs EF1 and EF2, RPB5f2 and RPB7cr. The sequences were deposited in GenBank [Accession TEF-1α: ON049647, RBP2: ON049648]. A Blast search of GenBank showed that TEF-1α and RPB2 sequences of SC03 matched Fusarium proliferatum (Matsush) Nirenberg at a high level (>99%). Phylogenetic analysis using neighbor joining and concatenated sequences (TEF-1α and RPB2) with MEGA X placed SC03 in F. proliferatum. For the pathogenicity test, a conidial suspension was prepared with a concentration of 2.0 × 107 conidia/ml. The suspension was sprayed onto 3 annual seedlings' needles, and the control was sprayed with sterile water. Inoculated and uninoculated plants were kept in humid chambers in a glasshouse where the average humidity was 60% and the average temperature was 27℃. After 10 days, typical symptoms appeared on inoculated needles, whereas control needles remained symptomless. These symptoms were similar to those observed in field. The fungus, F. proliferatum, was reisolated from those lesions, confirming Koch's postulates. No symptoms were observed on control plants. Fusarium proliferatum is a ubiquitous saprophytic fungus on cankers and very rarely reported to cause disease on pine needles. It has been reported to cause leaf blight of Polygonatum cyrtonema (Zhou et al. 2021) and Majesty palm (Polizzi and Vitale 2003). To our knowledge, this is the first report of needle blight on P. thunbergii caused by F. proliferatum in China. The disease represents a threat to producers and more research on the biology and management is needed.

Salix babylonica is an important landscape tree in China and has been widely planted. In this study, the pathogenicity of Septotinia populiperda causing leaf blotch of Sa. babylonica to four willow species ( Sa. matsudana , Sa. chaeomoloides , Sa. matsudana f. tortuosa , and Sa. suchowensis ) and Populus tomentosa (Chinese white poplar) was determined. Its sexual stage and biological characteristics were studied. Leaves from four willow species and P. tomentosa were inoculated with mycelial plugs. Typical leaf blotches with sporodochia were produced on all inoculated leaves. Among the isolates studied, some developed conidia but sclerotia were rare. The sclerotia developed apothecia after induction at 4°C for 3 months in an incubator and 2 more months outdoors from January to March. The biological characteristics of S. populiperda showed that mycelium grew better on complete medium than on potato dextrose agar, Czapek's agar, and minimal medium. For mycelial growth, the optimal carbon source was dextrose and the optimal nitrogen source was yeast powder. Conidia germination rate was 59.4% at 24 h. The conidia germinated best in a 4% willow leaf extraction. The optimal temperature for conidia germination was 25°C, and the optimal pH was 4.

Purpose: This study aims to evaluate the diagnostic application and performance of the metagenomic next-generation sequencing (mNGS) in patients suspected of local pulmonary infection by comparing it to the traditional pathogen detection methods in lung tissue specimens obtained by a computerized tomography-guided biopsy (CT-guided biopsy)., Methods: We retrospectively reviewed patients, admitted to the First Affiliated Hospital of Wenzhou Medical University, China from May 2018 to December 2020, who were suspected of local pulmonary infection. All cases received a CT-guided lung biopsy, tissue samples were sent both for conventional examinations (CE) and mNGS tests. The sensitivity and specificity of the two diagnostic approaches were compared., Results: 106 patients enrolled, 76 patients were diagnosed with a pulmonary infection. Among 49 patients with identified pathogens, CE confirmed pathogenic infections in 32 cases. Mycobacterium spp. and fungi accounted for 37.5% (12/32) and 28.1% (9/32), respectively, with bacteria 34.4% (11/32). The mNGS examination detected extra pathogenic microorganisms in 22 patients that were consistent with the patients' clinical and radiographic pictures. The sensitivity of mNGS was 53.9% vs. 42.1% for the CE, while the specificity was 56.7% versus 96.7%. For detection rate, mNGS was significantly superior to CE in bacterial (96.3% vs. 40.7%, p < 0.05), and mixed infections (100% vs. 50%, p < 0.05), but inferior to CE in fungal (60% vs. 90%, p > 0.05) and Mycobacterium spp. infections (66.7% vs. 100%, p > 0.05) with no significant difference. Among 31 cases diagnosed with lung abscess, the diagnostic performance of the detection rate was 67.7% (21/31) in favour of mNGS compared to 29.0% (9/31) for CE (p < 0.05). Most polymicrobial infections were induced by anaerobic species that coexisted with Streptococcus constellatus. And Klebsiella pneumoniae was the most common isolated monomicrobial infection., Conclusions: The most commonly detected causative pathogens for local pulmonary infections were bacteria, Mycobacterium spp. and fungi. Compared with the CE, the advantages of mNGS in the pathogens detection lie in the discovery of bacterial and mixed infections, as well as in the detection of lung abscess. Conversely, mNGS is not good enough to be recommendable for the detection of Mycobacterium spp. and fungi., (© 2022. The Author(s).)