A Dual-Cascade Activatable Near-Infrared Fluorescent Probe for Precise Intraoperative Imaging of Tumor.

Accurate intraoperative tumor delineation is critical to achieving successful surgical outcomes. However, conventional techniques typically suffer from poor specificity and low sensitivity and are time-consuming, which greatly affects intraoperative decision-making. Here, we report a cascade activatable near-infrared fluorescent (NIRF) probe IR780SS@CaP that can sequentially respond to tumor acidity and elevated glutathione levels for accurate intraoperative tumor localization. Compared with nonactivatable and single-factor activatable probes, IR780SS@CaP with a cascade strategy can minimize nonspecific activation and false positive signals in a complicated biological environment, affording a superior tumor-to-normal tissue ratio to facilitate the delineation of abdominal metastases. Small metastatic lesions that were less than 1 mm in diameter can be precisely identified by IR780SS@CaP and completely excised under NIRF imaging guidance. This study could benefit tumor diagnosis and image-guided tumor surgery by providing real-time information and reliable decision support, thus reducing the risk of both recurrence and complications to improve patient outcomes.

Altered interhemispheric functional connectivity in patients with obsessive-compulsive disorder and its potential in therapeutic response prediction.

The trajectory of voxel-mirrored homotopic connectivity (VMHC) after medical treatment in obsessive-compulsive disorder (OCD) and its value in prediction of treatment response remains unclear. This study aimed to investigate the pathophysiological mechanism of OCD, as well as biomarkers for prediction of pharmacological efficacy. Medication-free patients with OCD and healthy controls (HCs) underwent magnetic resonance imaging. The patients were scanned again after a 4-week treatment with paroxetine. The acquired data were subjected to VMHC, support vector regression (SVR), and correlation analyses. Compared with HCs (36 subjects), patients with OCD (34 subjects after excluding two subjects with excessive head movement) exhibited significantly lower VMHC in the bilateral superior parietal lobule (SPL), postcentral gyrus, and calcarine cortex, and VMHC in the postcentral gyrus was positively correlated with cognitive function. After treatment, the patients showed increased VMHC in the bilateral posterior cingulate cortex/precuneus (PCC/PCu) with the improvement of symptoms. SVR results showed that VMHC in the postcentral gyrus at baseline could aid to predict a change in the scores of OCD scales. This study revealed that SPL, postcentral gyrus, and calcarine cortex participate in the pathophysiological mechanism of OCD while PCC/PCu participate in the pharmacological mechanism. VMHC in the postcentral gyrus is a potential predictive biomarker of the treatment effects in OCD.

Improved fluorescence-based assay for rapid screening and evaluation of SARS-CoV-2 main protease inhibitors.

The outbreak of coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global threat to human health. In parallel with vaccines, efficacious antivirals are urgently needed. SARS-CoV-2 main protease (Mpro) is an attractive drug target for antiviral development owing to its key roles in virus replication and host immune evasion. Due to the limitations of currently available methods, the development of novel high-throughput screening assays is of the highest importance for the discovery of Mpro inhibitors. In this study, we first developed an improved fluorescence-based assay for rapid screening of Mpro inhibitors from an anti-infection compound library using a versatile dimerization-dependent red fluorescent protein (ddRFP) biosensor. Utilizing this assay, we identified MG-101 as a competitive Mpro inhibitor in vitro. Moreover, our results revealed that ensitrelvir is a potent Mpro inhibitor, but baicalein, chloroquine, ebselen, echinatin, and silibinin are not. Therefore, this robust ddRFP assay provides a faithful avenue for rapid screening and evaluation of Mpro inhibitors to fight against COVID-19.

Enhanced interhemispheric resting-state functional connectivity of the visual network is an early treatment response of paroxetine in patients with panic disorder.

This study aimed to detect alterations in interhemispheric interactions in patients with panic disorder (PD), determine whether such alterations could serve as biomarkers for the diagnosis and prediction of therapeutic outcomes, and map dynamic changes in interhemispheric interactions in patients with PD after treatment. Fifty-four patients with PD and 54 healthy controls (HCs) were enrolled in this study. All participants underwent clinical assessment and a resting-state functional magnetic resonance imaging scan at (i) baseline and (ii) after paroxetine treatment for 4 weeks. A voxel-mirrored homotopic connectivity (VMHC) indicator, support vector machine (SVM), and support vector regression (SVR) were used in this study. Patients with PD showed reduced VMHC in the fusiform, middle temporal/occipital, and postcentral/precentral gyri, relative to those of HCs. After treatment, the patients exhibited enhanced VMHC in the lingual gyrus, relative to the baseline data. The VMHC of the fusiform and postcentral/precentral gyri contributed most to the classification (accuracy = 87.04%). The predicted changes were accessed from the SVR using the aberrant VMHC as features. Positive correlations (p < 0.001) were indicated between the actual and predicted changes in the severity of anxiety. These findings suggest that impaired interhemispheric coordination in the cognitive-sensory network characterized PD and that VMHC can serve as biomarkers and predictors of the efficiency of PD treatment. Enhanced VMHC in the lingual gyrus of patients with PD after treatment implied that pharmacotherapy recruited the visual network in the early stages.

Evaluation of Stripe Rust Resistance and Chip Detection Resistance Genes in 286 Xinjiang Wheat Cultivars and Breeding Lines.

Wheat stripe rust is a destructive disease worldwide, caused by Puccinia striiformis f. sp. tritici (Pst). Resistance breeding is the most effective method of controlling stripe rust. Xinjiang is a relatively independent epidemic region of wheat stripe rust in China. In recent years, wheat stripe rust in this area has shown an upward trend. Therefore, the purpose of this study was to evaluate the resistance level of wheat cultivars (lines) to the prevalent Pst races and determine the genetic background of stripe rust resistance genes in Xinjiang. Six predominant Pst races in China were used to study resistance of 286 wheat cultivars (lines) at both seedling under controlled conditions and adult-plant stages under field conditions. In the seedling tests, 175 (61.19%) entries were resistant to races CYR23, 125 (43.71%) to CYR29, 153 (53.50%) to CYR31, 88 (30.77%) to CYR32, 174 (60.84%) to CYR33, and 98 (34.27%) to CYR34. Among the resistant entries, 23 (8.04%) were resistant to all six races. In the field test, 135 (47.20%) entries were resistant to the tested mixed races. Through comparing the responses in the seedling and adult-plant stages, 109 (38.11%) entries were found to have adult-plant resistance (APR), and 14 (4.90%) entries have all-stage resistance (ASR). The 286 wheat entries were also tested using a wheat breeder chip containing 12 Yr resistance loci. Among these entries, 44 (15.38%) were found to have single gene, 221 (77.27%) have two or more genes, and 21 (7.34%) have none of the 12 genes, including 144 (50.35%) with Yr30 and 5 (1.75%) with YrSP. Entries with two or more genes have stronger resistance to Pst. Overall, the majority of entries have all-stage and/or adult-plant resistance, but their genes for resistance in addition to the 12 tested Yr genes need to be determined. It is also necessary to introduce more effective resistance genes in the breeding programs to improve stripe rust resistance in wheat cultivars in Xinjiang.

First report of leaf spot disease caused by Neopestalotiopsis rosae on Lonicera caerulea L. in Heilongjiang Province, China.

Recently, interest in cultivating blue honeysuckle (Lonicera caerulea L.) for horticulture and medicinal uses has grown (Sharma and Lee 2021). Between September 2022 and September 2023, a leaf spot disease (Fig. S1) was observed on approximately 20% of 'Lanjingling' blue honeysuckles grown in a 0.18 ha field in Qiqihar city (123.43°E, 47.92°N), Heilongjiang Province, China. Infected plants displayed black leaf spots that expanded to cover the entire leaf. Small, 3 to 4 mm segments of infected tissue were surface sterilized with 75% ethanol for 30 s and 5% sodium hypochlorite (NaOCl) for 3 min, rinsed three times with sterile distilled water, dried on paper towels, and plated in 9 cm Petri dishes containing potato dextrose agar (PDA) (Ma et al. 2023). To induce sporulation, nine purified cultures (Fig. S2) with similar culture characteristics were finally obtained from ten infected plants and they displayed a conidia morphology consistent with Neopestalotiopsis spp., no other fungi were isolated, and the isolation frequency was 90%. Conidiomata (Fig. S3) were brown to black and distributed in concentric rings with an average size of 261.98 (60.30-451.80) µm (n = 50). The conidia (Fig. S3) were fusoid and had four septa, straight to slightly curved, with an average size of 23.48 (13.50-30.30) × 5.42 (4.50-9.30) µm(n = 50), while basal and apical cells were hyaline and the three middle cells were brown with darker septa. PCR amplification was performed with ITS1/ITS4 (White et al. 1990), EFl-728F/EF1-986R (Carbone and Kohn 1999), and Btub2Fd/Btub4Rd (Glass and Donaldson 1995) primers from the genomic DNA of the LD-330. Sequences of ITS (PP033584), TEF (PP048757), and TUB (PP048758) revealed 99 to 100% (499/500, 255/255, and 481/486) shared identity with Neopestalotiopsis rosae sequences (NR145243, KM199524, and KM199430) (Rebollar-Alviter et al. 2020). Therefore, based on morphological characteristics and molecular phylogeny, LD-330 was identified as N. rosae. Six two-year-old healthy plants of the 'Lanjingling' cultivar were selected for a pathogenicity test (Yan et al. 2023). The leaves were surface disinfected with 75% alcohol and then wiped with sterilized water three times. Three plants were inoculated with 10 ml of LD-330 conidial suspension (1 × 106 spores/ml) or with sterile water as an experimental control, respectively. All plants were in closed plastic bag, incubated in a greenhouse at 28 ℃ and 75% relative humidity (RH) under a 12-h light/dark cycle, and each experiment was performed three times (Rebollar-Alviter et al. 2020). Typical leaf spot symptoms were observed on inoculated leaves after 14 days (Fig. S4), whereas no symptoms were detected on water-treated leaves. The same pathogen was reisolated from infected leaves, displayed the same morphological and molecular traits, and was again identified as N. rosae, confirming Koch's postulate. Neopestalotiopsis rosae was previously reported on pecan (Gao et al. 2022), causing black leaf spot disease in China. To our knowledge, this is the first report of a blue honeysuckle leaf spot caused by N. rosae in China and specifically in the Heilongjiang province which has the largest blue honeysuckle cultivation area in the country. Future research should be directed toward developing comprehensive management measures.

First report of leaf spot disease caused by Pseudopithomyces chartarum on Lonicera caerulea L. in Heilongjiang Province, China.

Blue honeysuckle (Lonicera caerulea L.) cultivation has gradually expanded in China but continues to be limited by challenges such as leaf spot disease. Between September 2022 and September 2023, a leaf spot disease was observed on approximately 30% of 'Lanjingling' blue honeysuckles grown in a 2.66 ha field (a total of about 11,000 plants) in Jiamusi city (130.47°E, 46.16°N), Heilongjiang Province, China. Affected plants displayed brown necrotic lesions on their leaves that gradually expanded in area until the leaves fell off the plant entirely. Small, 3 to 4 mm segments of infected tissue from 50 randomly selected leaves were surface sterilized with 75% ethanol for 30 s and 5% sodium hypochlorite (NaOCl) for 3 min, rinsed three times with sterile distilled water, dried on paper towels, and plated in 9 cm Petri dishes containing potato dextrose agar (PDA) (Yan et al. 2022). Five pathogens (LD-232, LD-233, LD-234, LD-235, and LD-236) were isolated on PDA and displayed a conidia morphology consistent with Pseudopithomyces spp. (Perelló et al. 2017). The fungal colonies on PDA were villiform, white, and whorled and had sparse aerial mycelium on the surface with black conidiomata. The conidia were obpyriform and dark brown, had 0 to 3 transverse and 0 to 1 longitudinal septa, and measured 9.00 to 15.30 µm × 5.70 to 9.30 µm in size (n = 50). Genomic DNA was extracted from a representative isolate, LD-232, for molecular verification and PCR amplification was performed with ITS1/ITS4 (White et al. 1990), LROR/LR7 (Carbone and Kohn 1999), and RPB2-5F2/RPB2-7CR (Liu et al. 1999) primers. Sequences of LD-232 ITS (OR835654), LSU (OR835652), and RPB2 (OR859769) revealed 99.8% (530/531 nt), 98.8% (639/647 nt), and 99.8% (1015/1017 nt) shared identity with Pseudopithomyces chartarum sequences (OP269600, OP237014, and MK434892), respectively (Wu et al. 2023). Bayesian inference (BI) was used to construct the phylogenies using Mr. Bayes v. 3.2.7 to confirm the identity of the isolates (Ariyawansa et al. 2015). Phylogenetic trees cannot be constructed based on the genes' concatenated sequences because selective strains do not have complete rDNA-ITS, LSU, and RPB2 sequences. Therefore, based on the morphological characteristics and molecular phylogeny, LD-232 was identified as P. chartarum (Perelló et al. 2017; Wu et al. 2023). A pathogenicity test was performed with six healthy, two-year-old 'Lanjingling' blue honeysuckle plants. Three plants were inoculated by spraying the LD-232 conidial suspension (1 × 106 spores/ml) or clean water as an experimental control condition (Wu et al. 2023; Yan et al. 2023). All plants were cultured in a greenhouse at 28℃ under a 12-h light/dark cycle, and each experiment was replicated three times. Typical leaf spot symptoms were observed on inoculated leaves after 10 days. The same pathogens were reisolated from infected leaves, displayed the same morphological and molecular traits, and were again identified as P. chartarum, confirming Koch's postulate. P. chartarum previously caused leaf spot disease on Tetrapanax papyrifer in China (Wu et al. 2023). To our knowledge, this is the first report of blue honeysuckle leaf spot caused by P. chartarum in China. Identification of P. chartarum as a disease agent on blue honeysuckle will help guide future management of leaf diseases for this economically important small fruit tree.

A Novel Estrone Degradation Gene Cluster and Catabolic Mechanism in Microbacterium oxydans ML-6.

Global-scale estrone (E1) contamination of soil and aquatic environments results from the widespread use of animal manure as fertilizer, threatening both human health and environmental security. A detailed understanding of the degradation of E1 by microorganisms and the associated catabolic mechanism remains a key challenge for the bioremediation of E1-contaminated soil. Here, Microbacterium oxydans ML-6, isolated from estrogen-contaminated soil, was shown to efficiently degrade E1. A complete catabolic pathway for E1 was proposed via liquid chromatography-tandem mass spectrometry (LC-MS/MS), genome sequencing, transcriptomic analysis, and quantitative reverse transcription-PCR (qRT-PCR). In particular, a novel gene cluster (moc) associated with E1 catabolism was predicted. The combination of heterologous expression, gene knockout, and complementation experiments demonstrated that the 3-hydroxybenzoate 4-monooxygenase (MocA; a single-component flavoprotein monooxygenase) encoded by the mocA gene was responsible for the initial hydroxylation of E1. Furthermore, to demonstrate the detoxification of E1 by strain ML-6, phytotoxicity tests were performed. Overall, our findings provide new insight into the molecular mechanism underlying the diversity of E1 catabolism in microorganisms and suggest that M. oxydans ML-6 and its enzymes have potential applications in E1 bioremediation to reduce or eliminate E1-related environmental pollution. IMPORTANCE Steroidal estrogens (SEs) are mainly produced by animals, while bacteria are major consumers of SEs in the biosphere. However, the understanding of the gene clusters that participate in E1 degradation is still limited, and the enzymes involved in the biodegradation of E1 have not been well characterized. The present study reports that M. oxydans ML-6 has effective SE degradation capacity, which facilitates the development of strain ML-6 as a broad-spectrum biocatalyst for the production of certain desired compounds. A novel gene cluster (moc) associated with E1 catabolism was predicted. The 3-hydroxybenzoate 4-monooxygenase (MocA; a single-component flavoprotein monooxygenase) identified in the moc cluster was found to be necessary and specific for the initial hydroxylation of E1 to generate 4-OHE1, providing new insight into the biological role of flavoprotein monooxygenase.

Disrupted functional connectivity associated with cognitive impairment in generalized anxiety disorder (GAD) and comorbid GAD and depression: a follow-up fMRI study.

BACKGROUND: Impaired functional connectivity between the bilateral hemispheres may serve as the neural substrate for anxiety and depressive disorders, yet its role in comorbid generalized anxiety disorder (GAD) and depression, as well as the effect of treatment on this connectivity, remains unclear. We sought to examine functional connectivity between homotopic regions of the 2 hemispheres (voxel-mirrored homotopic connectivity [VMHC]) among people with GAD with and without comorbid depression at baseline and after a 4-week paroxetine treatment. METHODS: Drug-naïve patients with GAD, with or without comorbid depression and healthy controls underwent functional magnetic resonance imaging and clinical assessments at baseline and after treatment. We compared VMHC and seed-based functional connectivity across the 3 groups. We performed correlation analysis and support vector regression (SVR) to examine the intrinsic relationships between VMHC and symptoms. RESULTS: Both patient groups (n = 40 with GAD only, n = 58 with GAD and depression) showed decreased VMHC in the precuneus, posterior cingulate cortex and lingual gyrus compared with healthy controls (n = 54). Moreover, they showed decreased VMHC in different brain regions compared with healthy controls. However, we did not observe any significant differences between the 2 patient groups. Seeds from abnormal VMHC clusters in patient groups had decreased functional connectivity. Voxel-mirrored homotopic connectivity in the precuneus, posterior cingulate cortex and lingual gyrus was negatively correlated with cognitive impairment among patients with GAD only and among all patients. The SVR analysis based on abnormal VMHC showed significant positive correlations (p < 0.0001) between predicted and actual treatment responses. However, we did not observe significant differences in VMHC or functional connectivity after treatment. LIMITATIONS: A notable dropout rate and intergroup somatic symptom variations may have biased the results. CONCLUSION: Patients with GAD with or without comorbid depression exhibited shared and distinct abnormal VMHC patterns, which might be linked to their cognitive deficits. These patterns have the potential to serve as prognostic biomarkers for GAD.Clinical trial registration: ClinicalTrials.gov NCT03894085.

First report of leaf spot disease caused by Alternaria tenuissima on Lonicera caerulea L. in Heilongjiang Province, China.

Blue honeysuckle (Lonicera caerulea L.) is a perennial plant of the Caprifoliaceae family and Lonicera genus, the largest genus in the plant kingdom. Between September 2021 and September 2022, a leaf spot disease was observed on ~20% of blue honeysuckles of the 'Lanjingling' cultivar grown in a 3.33 ha field at the Xiangyang base (126.96°E, 45.77°N) of the Northeast Agricultural University, Harbin (Heilongjiang Province, China). Leaf spots first presented black mildew centers, gradually covering large areas of the leaf until it eventually fell off. Small 3-4 mm segments of infected tissue from 50 randomly selected leaves were surface sterilized with 75% ethanol and 5% sodium hypochlorite, rinsed in sterile distilled water, and transferred to 9 cm Petri dishes containing potato dextrose agar (PDA) after drying. Finally, two isolated pathogens were obtained through single spore culture on PDA; they appeared as gray-black colonies and were named LD-12 and LD-121. The observed LD-12 and LD-121 conidia displayed a morphology consistent with Alternaria spp. They were obpyriform and dark brown, with 0-6 transverse and 0-3 longitudinal septa, measuring 6.00-17.70 µm × 9.30-42.30 µm and 5.70-20.70 µm × 8.40-47.70 µm for LD-12 and LD-121, respectively (n = 50). Genomic DNA was extracted from the two isolates for molecular verification, and PCR amplification was performed with ITS1/ITS4 (White et al. 1990), GPD1/GPD2 (Woudenberg et al. 2015), EFl-728F/EF1-986R (Carbone and Kohn 1999), RPB2-5F2/RPB2-7CR (Liu et al. 1999), and Alt-for/Alt-rev (Hong et al. 2005) primers. Sequences of LD-12 ITS (OQ607743), GPD (OQ623200), TEF (OQ623201), RPB2 (OQ658509), and ALT (OQ623199) revealed 99-100% of identity with Alternaria tenuissima sequences (KC584567, MK451973, LT707524, MK391051, and ON357632). Sequences of LD-121 ITS (OQ629881), GPD (OQ850078), TEF (OQ850075), RPB2 (OQ850076), and ALT (OQ850077) revealed 99-100% identity with A. alternata sequences (MN826219, ON055384, KY094927, MK637444, and OM849255). Nine two-year-old healthy plants from the 'Lanjingling' cultivar were selected for a pathogenicity test. Three plants were inoculated with either the LD-12 or LD-121 conidial suspension (1 × 106 spores/ml) or with clean water as an experimental control condition (Mirzwa-Mróz et al., 2018; Liu et al., 2021). All plants were cultured in a greenhouse at 28℃ under a 12-h light/dark cycle, and each experiment was performed three times. Typical leaf spot symptoms were observed on inoculated leaves after 10 d. The same pathogens reisolated from infected leaves displayed the same morphological and molecular traits. They were again identified as A. tenuissima and A. alternata, confirming Koch's postulate. A. tenuissima and A. alternata were previously reported on Orychophragmus violaceus (Liu et al., 2021) and L. caerulea (Yan et al., 2022) in China. This study is the first report of a blue honeysuckle leaf spot caused by A. tenuissima in China. In the future, effective biological and chemical control should be used to prevent blue honeysuckle leaf spots in China.

First report of Didymella glomerata causing leaf spot on Lonicera caerulea L. in Heilongjiang province, China.

China has the largest blue honeysuckle (Lonicera caerulea L.) cultivation area globally. In June 2022, leaf spots were observed on approximately 10% of blue honeysuckle (cv. 'Lanjingling') leaves in a 0.03-ha field in Harbin (127.66°E, 45.61°N), Heilongjiang Province, China. The leaves of the affected plants displayed chlorotic to tan dieback with a darker brown margin along the leaftip and leave margins. Cross-sectional segments of approximately 3 mm were cut from 50 typical infected plant leaves. Their surfaces were sterilized with 75% ethanol for 30 s followed by 3 min in 5% sodium hypochlorite (NaOCl), rinsed three times with sterile water, and transferred to 9-cm Petri dishes containing 15 ml of sterile PDA growth medium. Five purified cultures with similar culture characteristics were finally obtained and their colonies were dark brown on the PDA plates. The pycnidia were subglobular and deep black and measured avg. 215.48 (135.30-331.20) µm × avg. 170.28 (99.90-282.90) µm (n = 50) (Chen et al., 2015; Huang et al., 2018). Conidia were single-celled, hyaline, and ellipsoidal and measured avg. 6.22 (5.40-7.20) µm × avg. 3.42 (2.70-3.90) µm (n = 50). For molecular verification, genomic DNA was extracted from a representative isolate, LD-75. The internal transcribed spacer region (ITS), the second-largest subunit of RNA polymerase II (rpb2), the partial 28S large subunit rDNA (LSU), beta-tubulin (TUB), and actin (ACT) genes were amplified with the primers ITS1/ITS4, RPB2f/RPB2r, LROR/LR7, TUB2Fd/TUB4Rd, and ACT512f/ACT783R, respectively (White et al. 1990; Carbone and Kohn, 1999; Staats et al., 2005; de Gruyter et al., 2009; Chen et al., 2015). BLAST results indicated that the genes of LD-75 (GenBank OP218870, OP264863, OQ561448, OQ597233, and OQ597232) shared 99%-100% identity with those of Didymella glomerata (OK485138, GU371781, EU754185, MZ073910, and MW963190, respectively). Therefore, based on morphological characteristics and molecular phylogeny, LD-75 was identified as D. glomerata. Six two-year-old healthy plants from the 'Lanjingling' cultivar were selected for a pathogenicity test. The leaves were surface disinfested with 75% ethanol and then wiped with sterilized water three times. All plants were cultured in a greenhouse at 28℃ under a 12-h light/dark cycle. Whole plants sprayed with conidial suspension of isolate LD-75 (106 spores/mL) (n = 3) displayed leaf spot symptoms after 14 d, while no symptoms were detected on whole plants sprayed with sterile water (n = 3). The same isolate, reisolated from infected leaves and with the same morphological and molecular traits, was also identified as D. glomerata, confirming Koch's postulate. The fungus was previously reported in Cornus officinalis in Nanyang City, China (Huang et al., 2018). To our knowledge, this is the first report of blue honeysuckle leaf spot caused by D. glomerata in China. Reducing blue honeysuckle production losses caused by leaf spots is crucial for growers, and we hope that researchers will develop efficient control strategies for managing this emerging plant disease.

First Report of Postharvest Fruit Rot Caused by Botrytis cinerea on Blue Honeysuckle (Lonicera caerulea L.) Fruit in China.

Blue honeysuckle (Lonicera caerulea L.) fruit is growing in popularity as a natural, functional 'super fruit', but its storage is challenged by pathogen infection. In June 2022, approximately 30% of 100 kg of blue honeysuckle fruits (cv. Lanjingling) obtained in Harbin, China (128.70°E, 44.87°N) showed postharvest fruit rot symptoms after 15 d of storage at 4°C, leading to whole fruit rotting with gray fungal growth (Fig.1 A). Small (1-2 mm) segments of infected tissue were obtained from 20 randomly selected fruits which were surface sterilized with 75% ethanol for 30 s and 5% sodium hypochlorite (NaOCl) for 3 min, rinsed three times with sterile distilled water, dried in paper towel, and plated in 9 cm Petri dishes containing potato dextrose agar (PDA). Five purified cultures were obtained and their front colonies were dark brown (Fig.1 C) on the PDA plates after 5 d at 25°C (Alam et al. 2019; Riquelme-Toledo et al. 2020). The conidia (n = 50) were single-celled, hyaline, either ellipsoid or ovoid, and measured 7.5-15.0 µm (11.7 µm average) × 6.0-11.4 µm (8.3 µm average). The conidiophores (Fig.1 E) were branched at the apex bearing bunches of conidia resembling grape clusters (Ellis 1971). For molecular confirmation, genomic DNA was extracted from a representative isolate LDGS-3 using the Ezup Column Fungi Genomic DNA Purification kit (Sangon Biotech, Shanghai, China). The internal transcribed spacer region (ITS, GenBank ON952502), heat shock protein (HSP60, GenBank OP039103), the second-largest subunit of RNA polymerase II (RPB2, GenBank OP186114) and glyceraldehyde 3-phosphate dehydrogenase (G3PDH, GenBank OQ658508) genes were partially amplified with the respective primers ITS1/ITS4, HSP60f/HSP60r, RPB2f/RPB2r, and G3PDH-F/G3PDH-R (Staats et al. 2005; White et al. 1990). BLAST analysis revealed that the sequences of the four genes showed 100% homology with the MH782039, MH796663, MN448501 and MH796662 sequences for isolates of Botrytis cinerea. Based on morphology and molecular characteristics, the isolate LDGS-3 was identified as B. cinerea. For pathogenicity, twenty healthy blue honeysuckle fruits (cv. Lanjingling) were superficially sterilized with 75% ethanol and washed with distilled water. Ten inoculated blue honeysuckle fruits, which were injected with 10 µL conidial suspension of isolate LDGS-3 (106 spores/mL) displayed fruit rot symptoms (Fig.1 B) inside 9 cm Petri dishes after 10 d at 4°C, while no symptoms were detected on ten fruits inoculated with sterile distilled water (Alam et al. 2019). The same isolate that was reisolated from infected fruits with the same morphological and molecular traits was also identified as B. cinerea, confirming Koch's postulates. B. cinerea was previously reported in Henan Province, China in hawthorn (Zhang et al. 2018). To our knowledge, this is the first report of postharvest fruit rot caused by B. cinerea on blue honeysuckle fruit in China, which will aid future management of this emerging postharvest disease.

An Activatable Near-Infrared Fluorescent Probe for Precise Detection of the Pulmonary Metastatic Tumors: A Traditional Molecule Having a Stunning Turn.

An accurate detection of lung metastasis is of great significance for making better treatment choices and improving cancer prognosis, but remains a big challenge in clinical practice. In this study, we propose a reinventing strategy to develop a pH-activatable near-infrared (NIR) fluorescent nanoprobe, pulmonary metastasis tracer (denoted as PMT), based on assembly of NIR dye IR780 and calcium phosphate (CaP). By delicately tuning the intermolecular interactions during the assembly process and dye doping content, as well as the synthetic condition of probe, the fluorescence of PMT could be finely adjusted via the tumor acidity-triggered disassembly. Notably, the selected PMT9 could sharply convert subtle pH variations into a distinct fluorescence signal to generate high fluorescence ON/OFF contrast, dramatically reducing the background signals. Benefiting from such preferable features, PMT9 is able to precisely identify not only the tumor sites in orthotopic lung cancer models but also the pulmonary metastases in mice with remarkable signal-to-background ratio (SBR). This study provides a unique strategy to turn shortcomings of traditional dye IR780 during in vivo imaging into advantages and further expand the application of fluorescent probe to image lung associated tumor lesions.

First report of leaf spot disease caused by Alternaria alternata on Lonicera caerulea L. in China.

Relatively few studies investigating plant diseases in blue honeysuckle (Lonicera caerulea L.) have been conducted in China. In September 2021, a leaf spot disease was observed on approximately 10% of blue honeysuckle 'Lanjingling' leaves in a 0.02 ha field plantation in Tiekuang Street (124.36°E, 40.12°N) in Dandong City, Liaoning Province, China. The main symptom consisted of leaf spots with black mildew centers typically surrounded by yellow halos. Small pieces (3-4 mm) of the infected leaves were plated onto potato dextrose agar (PDA) medium as described by Wang et al. (2020) and six purified cultures were obtained through single spore culture on PDA. The observed conidia, consistent with the morphology of Alternaria alternata, were obpyriform and dark brown, measuring 5.8 to 15.3 µm × 7.9 to 42.5 µm, with 1-6 transverse septa and 0-3 longitudinal septa (n = 50) (Simmons 2007). For molecular verification, genomic DNA was extracted from a representative isolate LD-8. The ITS (GenBank OL454815), GPD (GenBank OL601993), TEF (GenBank OL538256), RPB2 (GenBank OL601966), and Alt (GenBank OL538257) genes were partially amplified with the respective primers ITS1/ITS4 (White et al. 1990), GPD1/GPD2 (Woudenberg et al. 2015), EFl-728F/EFI-986R (Carbone and Kohn 1999), RPB2-5F2/RPB2-7CR (Liu et al. 1999), and Alt-for/Alt-rev (Hong et al. 2005). BLAST analysis revealed that these genes shared 99%-100% identity with OK345332, MK451977, MN756011, KU933459, and MN655781, respectively. A greenhouse experiment was conducted using six, healthy two-year-old blue honeysuckle 'Lanjingling' plants to observe disease development (Mirzwa-Mróz et al. 2018). After 10 d, we noted typical leaf spot symptoms on inoculated leaves sprayed with a conidial suspension (106 spores/mL) while no symptoms were detected on uninoculated leaves. The same isolate, reisolated from infected leaves with the same morphological and molecular traits, was also identified as A. alternata, confirming Koch's postulates. The fungus was previously reported in cockscomb plants in Heilongjiang Province, China (Wang et al. 2020). To our knowledge, this is the first report of leaf spot disease caused by A. alternata in blue honeysuckle grown in China. This study will provide a basis for future development of effective protection strategies against blue honeysuckle leaf spot in China.

Mental Health of Medical Staff During the Coronavirus Disease 2019 Pandemic: A Systematic Review and Meta-Analysis.

OBJECTIVE: This study aimed to quantify the prevalence of the adverse mental health outcomes in medical staff working in the hospital settings during the coronavirus disease 2019 (COVID-19) pandemic and explore the relative distribution of anxiety and depressive symptoms. METHODS: PubMed, EMBASE, China National Knowledge Infrastructure, WANFANG DATA, and VIP Database for Chinese Technical Periodicals were searched for articles published from January 1, 2019, to April 19, 2020. The prevalence estimates of adverse mental health symptoms in medical staff were pooled using the random-effects model. RESULTS: A total of 35 articles and data of 25,343 medical staff were used in the final analysis. The pooled prevalence estimates in medical staff during the COVID-19 pandemic were as follows (ordered from high to low): fear-related symptoms, 67% (95% confidence interval [CI] = 61%-73%); high levels of perceived stress, 56% (95% CI = 32%-79%), anxiety symptoms, 41% (95% CI = 35%-47%); insomnia, 41% (95% CI = 33%-50%); posttraumatic stress disorder symptoms, 38% (95% CI = 34%-43%); depressive symptoms, 27% (95% CI = 20%-34%); and somatic symptoms, 16% (95% CI = 3%-36%). The subgroup analysis revealed that the prevalence estimates of fear-related symptoms were consistently high. CONCLUSIONS: Medical staff during the COVID-19 epidemic have a high prevalence of adverse mental health symptoms. Data-based strategies are needed to optimize mental health of medical staff and other health care professionals during times of high demand such as the COVID-19 and other epidemics.PROSPERO Registration: CRD42020182433.

Antagonistic Activity of Bacillus velezensis SDTB038 against Phytophthora infestans in Potato.

Potato late blight is a severe and highly epidemic disease caused by Phytophthora infestans that can affect all parts of the plant. This study mainly screened antagonistic strains for good control of potato late blight and identified strain SDTB038 as Bacillus velezensis according to its morphological and chemical properties and the 16S rRNA, gyrA, and gyrB gene sequences. This antagonistic strain achieved good control of potato late blight in greenhouses and fields and promoted potato plant growth. Two-year field trials (2018 and 2019) showed that B. velezensis SDTB038 can be used to reduce food losses caused by late blight, achieving late blight reductions of 40.79% (2018) and 37.67% (2019). In two-year field trials, the control effects of the highest concentrations of fluopimomide and B. velezensis SDTB038 were better than those of the other treatments. The control effect of 85 g ha-1 fluopimomide and B. velezensis SDTB038 and that of 170 g ha-1 fluopimomide alone showed no significant differences. These field results indicate that a low concentration of fungicide and a high concentration of SDTB038 can be effective in controlling potato late blight. Foliar detection showed that lipopeptides have an inhibitory effect on P. infestans. The amplification of lipopeptide genes revealed surfactin (srfAB and srfAC) and fengycin (fenB) genes in SDTB038, but only surfactin production by B. velezensis SDTB038 was observed by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry analysis. Therefore, the strain B. velezensis SDTB038 can produce secondary metabolites that help potato plants resist late blight development, can effectively inhibit the infection of potato leaves by P. infestans, and has potential value for development as a biological pesticide against potato late blight.