Knowledge, Attitudes, and Practice of General Practitioners Toward Screening of Age-Related Hearing Loss in Community: A Cross-Sectional Study in Shanghai, China.

Background: Age-related hearing loss (ARHL) is experiencing a continuously rising in prevalence among the elderly worldwide. General practitioners (GPs) may have a unique position in its community detection and management. Objective: This study aims to assess the KAP of GPs regarding ARHL through questionnaire, to investigate the role of them in the management and to propose strategies for the hearing screening within the community. Methods: An online survey was administered to 1173 GPs, selected from 56 community health centers (CHCs) in Shanghai during April to June 2022. A scale endorsed by a panel of multidisciplinary experts was used to assess knowledge (7 items), attitudes (12 items), and practice (10 items). A mean score was computed and converted into a scale ranging from 0 to 100. Odds ratios (ORs) were calculated for potential predictors of higher levels of KAP scores (with mean value as a cutoff point) through logistic modelling. Results: A total of 1022 GPs completed the questionnaire with response rate 87.13%. The average scores are 69.90 ± 32.27, 66.09 ± 7.15, and 59.89 ± 21.99 for Knowledge, attitude, and practice, respectively. 24.3% of participants achieve a complete score of knowledge, whereas 5.48% receive zero. 11.6% consider ARHL as not a disease. Above 30.0% are not familiar with the screening tool. 10.8% refuse to undergo hearing screening. Higher levels of compliance in practice are found in the participants with higher levels of knowledge (OR=1.409, p=0.000) and more favorable attitude (OR=1.028, p=0.000). Male (OR=0.708, p=0.036) is associated with lower levels of attitudes. Conclusion: GPs have a low level of ARHL knowledge, a lack of positive attitude towards the detection and management of it, and lower awareness in practice. Further research is required to gain a more comprehensive understanding of the attitudes held by GPs and explore more accessibility strategies.

Pulmonary tumor embolism in a maintenance hemodialysis patient with hepatocellular carcinoma.

Patients with chronic kidney disease are already at an increased risk for pulmonary embolism, since loss of renal function rendered a procoagulant state. Further, malignant tumor is a well-established risk factor for pulmonary thromboembolism. Alternatively, occlusion of the pulmonary vasculature by tumor cells per se and associated thrombi may mimic thromboembolic disease. By comparison, however, report of pulmonary tumor embolism (PTE) in patients on maintenance hemodialysis (MHD) is exceedingly rare. A less vigilant clinician may have otherwise treated this situation as fluid overload or thromboembolic disorder. We herein described in an MHD patient such an unusual case of PTE, which was diagnosed by contrast-enhanced CT and PET/CT. As such, our work may expand the knowledge reserve of dialysis staffs about this rare complication of malignancy.

Pulmonary cryptococcosis closely mimicking lung cancer in a membranous nephropathy patient taking calcineurin inhibitor.

In patients with membranous nephropathy (MN), malignancy may be either the underlying disease or results of immunosuppressive therapy which may also lead to opportunistic infections including the pulmonary cryptococcosis. On CT scan, nodule is the most common feature in pulmonary cryptococcosis and it can mimic lung cancer both clinically and radiologically. Therefore, pulmonary nodular lesions caused by cryptococcosis may be easily misdiagnosed and require unnecessary surgical treatment. As such, we herein presented an isolated subpleural solitary nodule with satellite lesion that closely mimicked lung cancer on both contrast-enhanced computed tomography (CT) scan and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET)/CT in an MN patient under long-term tacrolimus regimen. Cryptococcosis was ascertained by the finding of oval thick-walled yeast on histopathology of the lung biopsy specimen taken during the Argon-Helium cryotherapy. Further, the pulmonary lesions progressively dissipated after antifungal treatment. Arguably, our experience may help clinicians in general and nephrologists in particular with a better understanding of the cryptococcal infection manifesting as pulmonary nodule(s) in the MN patients and contribute to more efficacious differential diagnosis against the lung cancer.

Hemophagocytic lymphohistiocytosis and histiocytic necrotizing lymphadenitis secondary to hemodialysis catheter-related bloodstream infection caused by Corynebacterium Striatum.

BACKGROUND: We herein described the coexistence of hemophagocytic lymphohistiocytosis (HLH) and histiocytic necrotizing lymphadenitis, alternatively known as the Kikuchi disease (KD), secondary to hemodialysis catheter-related bloodstream infection (BSI) caused by Corynebacterium striatum. CASE PRESENTATION: A patient on maintenance hemodialysis had developed persistent fever and Corynebacterium striatum was subsequently identified from the culture of both catheter tip and peripheral blood. During mitigation of the BSI, however, his fever was unabated and ensuing workup further found thrombocytopenia, hyperferritinemia, hypertriglyceridemia, low NK cell activity and a surge in serum CD25 levels. Moreover, biopsy of the bone marrow and lymph node detected histopathological evidence of hemophagocytosis and KD, respectively. Upon these abnormalities, the title-bound diagnosis was considered and the patient was eventually recovered from the treatment of dexamethasone instead of antibiotics. Consistently, aberrations in his serum CD25 levels and NK cell activity had subsided two months after discharge. CONCLUSIONS: Arguably, this encounter offered a unique chance to unravel the principal pathogenic cascade in immunobiology that made the three entities one disease continuum. As such, our work may add new understandings of HLH and/or KD secondary to severe infections in general and excessive release of cytokines in particular among patients with kidney diseases. The resultant early diagnosis is crucial to initiate appropriate treatment and improve the survival of patients with these challenging and potentially life-threatening disorders.

Construction and Effect of the Three-Level and Two-Stage Screening Mode for Age-Related Hearing Loss: A Study Based on the Community in Shanghai, China.

Background: There is a high incidence rate of age-related hearing loss. Severe hearing loss may increase the prevalence of mental illness, cognitive impairment, and even the risk of all-cause death. Purpose: Construction of the three-level and two-stage screening mode for age-related hearing loss of the community and to evaluate its effectiveness. Materials and Methods: A total of 401 participants (aged 60 years or older) from five typical communities were enrolled in the study. The risk factors assessment of age-related hearing loss was completed by using a cross-sectional survey and receiver operating characteristic (ROC) curve. Multiple screening method was adopted and verified by serial and parallel tests, respectively. Based on research data, incorporate risk factors assessment, the Hearing Handicap Inventory for the Elderly Screening Version (HHIE-s) and pure tone audiometry (PTA) were used to construct the screening mode. Results: Multiple screening series testing and multiple screening parallel testing, including risk factors assessment, HHIE-s, and PTA, were used for verification: the sensitivity, specificity, and Kappa index were 70.5% and 9.2%, 95.0% and 71.6%, 0.26 and 0.63, respectively. Finally, the three-level and two-stage screening mode for age-related hearing loss was established. "Three-level" was defined as the risk factors assessment/HHIE-s (high-risk population), PTA (suspect population), and comprehensive hearing loss assessment (confirmed population). "Two-stage" was defined as the population screening by general practitioner in the community and target screening by otolaryngologist of the tertiary hospitals. Conclusion: The three-level and two-stage screening mode for age-related hearing loss consists of the following framework: from population screening to target screening, from suspicious diagnosis to accurate diagnosis, from primary health care to tertiary hospitals. The study objective is to structure a new secondary prevention and treatment mode for age-related hearing loss with primary health care as the core, so as to help the front-end management of healthy aging.

Delivery of Methoprene-Tolerant dsRNA to Improve RNAi Efficiency by Modified Liposomes for Pest Control.

The application of RNA interference (RNAi) technology for pest control is environmentally friendly and accurate. However, the efficiency of RNAi is often inconsistent and unreliable, and finding a suitable carrier element is considered critical to success in overcoming biotic and abiotic barriers to reach the target site. The fall armyworm, Spodoptera frugiperda (FAW), which is one of most important global agricultural pests, has recently spread rapidly to other parts of the world. In this study, a method to improve the stability and RNAi efficiency of the dsRNA carrier complex was reported. Methoprene-tolerant gene (Met) was selected as a target, a gene which is critical to the growth and development of FAW. Biomaterials nanoliposomes (LNPs) were modified with polyethylenimine (PEI) to deliver the dsRNA of Met. The synthesized Met3@PEI@LNPs reached a size of 385 nm and were found to load dsRNA effectively. Through stability and protection assays, it was found that LNPs provided reliable protection. In addition, the release curve also demonstrated that LNPs were able to prevent premature release under alkaline condition of the insect midgut but accelerate the release after entering the acidic environment of the target cells. The cell transfection efficiency of the prepared LNPs reached 96.4%. Toxicity tests showed that the use of LNPs could significantly improve the interference efficiency, with 91.7% interference efficiency achieved when the concentration of dsRNA in LNPs was only 25% of that of the control. Successful interference of Met demonstrated it could significantly shorten the larval period and make the larvae pupate earlier, thus achieving the purpose of control. In this study, we have demonstrated the use of nanotechnology to provide a novel RNAi delivery method for pest control.

A novel GJA5 variant associated with increased risk of essential hypertension.

OBJECTIVES: Gap junction protein alpha 5 (GJA5), also termed connexin 40 (Cx40), exerts a pivotal role in the mediation of vascular wall tone and two closely-linked polymorphisms in the GJA5 promoter (-44G>A and +71A>G) have been associated with enhanced susceptibility to essential hypertension (EH) in men. The present investigation aimed to ascertain whether a novel common polymorphism within the upstream regulatory region of GJA5 (transcript 1B), -26A>G (rs10465885), confers an increased risk of EH. METHODS: For this investigation, 380 unrelated patients with EH and 396 unrelated normotensive individuals employed as control persons were enrolled from the Chinese Han-ethnicity population, and their GJA5 genotypes and plasma renin concentrations were determined by Sanger sequencing and an automated chemiluminescent immunoassay, respectively. The functional effect of the GJA5 variant was explored in cultured murine cardiomyocytes by dual-light reporter gene analysis. RESULTS: The GJA5 variant conferred a significantly increased risk for EH (OR: 2.156; 95% CL: 1.661-2.797, P < 0.0001), and significantly increased plasma renin levels were measured in patients with EH in comparison with control individuals (46.3±7.2 vs 37.4±6.9, P < 0.0001). A promoter-luciferase analysis revealed significantly diminished activity of the promoter harboring the minor allele for this variation in comparison with its wild-type counterpart (165.67±16.85 vs 61.53±8.67, P = 0.0007). CONCLUSIONS: These findings indicate that the novel variant upstream of the GJA5 gene (-26A>G) confers a significantly increased vulnerability of EH in humans, suggesting potential clinical implications for precisive prophylaxis and treatment of EH.

Circ_0008529 Contributes to Renal Tubular Cell Dysfunction in High Glucose Stress via miR-185-5p/SMAD2 Pathway in Diabetic Nephropathy.

Circular RNA (circRNA) is key regulator of diabetic nephropathy (DN) progression. However, the role of circ_0008529 in DN progression remains to be better deciphered. Cell viability, cell cycle, apoptosis and inflammation were measured by MTS assay, flow cytometry and corresponding assay kits. RT-qPCR was used to assess the expression of circ_0008529, miR-185-5p and SMAD family member 2 (SMAD2). Also, western blotting was performed to measure protein expression. Target relationship was validated by RNA pull-down assay, dual-luciferase reporter assay and RNA immunoprecipitation assay. Urinary exosome was isolated using ultracentrifugation method and identified by transmission electron microscopy. Receiver operating characteristic curve was used to analyze the diagnostic value of circ_0008529 in DN patients. Circ_0008529 and SMAD2 were upregulated, while miR-185-5p was downregulated in high glucose (HG)-induced renal tubular HK-2 cells. Under HG treatment, cell viability and cell cycle process were suppressed, while apoptosis, inflammation and extracellular matrix accumulation were enhanced. However, interfering circ_0008529 could attenuate HG-induced effects, and this protection was abated by miR-185 inhibition or SMAD2 re-expression. Mechanically, circ_0008529 and SMAD2 were competing endogenous RNAs for miR-185-5p via target binding, and circ_0008529 regulated SMAD2 expression via miR-185-5p. Notably, circ_0008529 expression was upregulated in urinary exosomes of DN patients, and showed diagnostic value (Sensitivity: 70.21%; Specificity: 86.67%). Circ_0008529 might be a potential target for DN, which regulated DN progression via miR-185-5p/SMAD2 pathway.

Screening of Insecticidal and Antifungal Activities of the Culturable Fungi Isolated from the Intertidal Zones of Qingdao, China.

Numerous studies focused on drug discovery perspective have proved the great potential for exploration of marine-derived fungi to seek bioactive chemicals. Yet, marine-derived fungi are less explored compared to their terrestrial counterparts. Here, 181 fungal strains (134 species) isolated from marine algae and sediment in Chinese intertidal zones were screened to reveal bioactivities using brine shrimp, green peach aphid and plant pathogens as targets. Fermentation supernatants of 85 fungal strains exhibited a high lethality (>70%) of brine shrimp at 24 h, and 14 strains appeared to be acute-toxic as featured by more than 75% mortality at 4 h, indicating efficient insecticidal bioactivity. The crude extracts of 34 strains displayed high toxicity to green peach aphid with more than 70% of mortality at 48 h. For the plant pathogens tested, the inhibitory rates of eight fungal strains affiliated with Alternaria (AS3, AS4), Amphichorda (AS7), Aspergillus (AS14), Chaetomium (AS21), Penicillium (AS46), Purpureocillium (AS55) and Trichoderma (AS67) were equal or higher than that of the positive Prochloraz, and five of them (AS7, AS14, AS21, AS55, AS67) were also strongly toxic to brine shrimp or aphid. Our findings indicate broad potential for exploration of marine-derived fungi as candidate resources to pursue bioactive compounds in controlling agricultural pests and pathogens.

Scoop thrombectomy: A declotting technique for the treatment of thrombosed autologous arteriovenous fistula. A single-center retrospective study.

BACKGROUND: Thrombosis is one of the main complications leading to the failure of autologous arteriovenous fistula (AVF) for patients with renal failure. Thrombectomy is one of the major therapies to remove thrombi to salvage the AVF and prolong its patency. MATERIALS AND METHODS: Fifty-six patients with AVF thrombosis at the anastomosis were recruited for this study and underwent thrombectomy procedures. Their clinical variables were collected. The vasculature was accessed at the site of the aneurysmal dilatation. Under ultrasound guidance, a scoop thrombectomy procedure was performed by anterograde and retrograde scooping to remove the thrombus using forceps. Then, a sheath was placed in the direct vertical direction. Angioplasty was performed with a balloon to treat the underlying primary arteriovenous stenosis. Patients were followed up for 12 months after surgery. The procedural success, primary and secondary patency rates, and incidence of procedure-related complications were analyzed. RESULTS: There were 2 minor (3.6%) and no major complications. Clinical success was achieved in 55 of the 56 procedures (98.2%). No symptomatic pulmonary embolism or arterial embolization was noted. The primary patency rates at 3, 6, and 12 months were 92.9, 83.8, and 73.3%%, respectively, according to the Kaplan-Meier survival analysis. CONCLUSION: Scoop thrombectomy is a safe procedure with high technical success and a low complication rate, and it is an effective method for patients to receive hemodialysis immediately.

Suppression of lncRNA GAS6-AS2 alleviates sepsis-related acute kidney injury through regulating the miR-136-5p/OXSR1 axis in vitro and in vivo.

Acute kidney injury (AKI) is a common complication of sepsis and increase morbidity and mortality. Long non-coding RNA (LncRNA) GAS6-AS2 was related to inflammation and apoptosis in different diseases by regulating miRNAs and downstream genes, but its role in AKI remains unclear. Thus, we speculated that GAS6-AS2 might function in sepsis-related AKI via regulating target genes. Here, LPS or CLP was used to establish in vitro or in vivo sepsis-related AKI model. The interactions between GAS6-AS2 and miR-136-5p, and miR-136-5p and OXSR1, were validated by luciferase reporter assay, RNA pull-down, or RIP assay. Cell apoptosis was determined by flow cytometry, Western blotting, or IHC. The kidney injury was evaluated by H&E staining. The expression of GAS6-AS2, miR-136-5p, and OXSR1 was determined by qRT-PCR or Western blotting. We found that GAS6-AS2 was up-regulated in LPS-treated HK2 cells and the CLP-induced rat model. In vitro, GAS6-AS2 knockdown decreased cleaved caspase-3 and bax expression and increased bcl-2 expression. The levels of TNF-α, IL-1ß, and IL-6 were reduced by GAS6-AS2 down-regulation. GAS6-AS2 knockdown ameliorated oxidative stress in the cells, as indicated by the reduced ROS and MDA levels and the elevated SOD level. In vivo, GAS6-AS2 down-regulation decreased urinary NGAL and Kim-1 levels and serum sCr and BUN levels, and H&E proved that the kidney injury was alleviated. GAS6-AS2 knockdown also reduced apoptosis, inflammation, and oxidation induced by CLP in vivo. Mechanically, GAS6-AS2 sponged miR-136-5p which targeted OXSR1. Overall, lncRNA GAS6-AS2 knockdown has the potential to ameliorate sepsis-related AKI, and the mechanism is related to miR-136-5p/OXSR1 axis.

First Report of Bacterial Leaf Spot on Tobacco Caused by Pseudomonas psychrotolerans in China.

Tobacco (Nicotiana tabacum) is an economically important crop, and its productivity is challenged due to pathogen infection. In 2020 and 2021, a previously uncharacterized disease was observed on field grown tobacco (Variety NC102) in Zhucheng City, Shandong Province, China (119°7'14" E, 36°0'58" N), where tobacco has been grown for decades. The disease can be found throughout the growth period of tobacco and mainly occurred from fast growing period (about 13-16 leaves) to leaf maturity stage. In severely diseased areas, the incidence rate can reach 100%. The symptoms first began as chlorotic water stain like small spots, then the spots merged into larger irregular necrotic maculae around the chlorotic halos. Small pieces of symptomatic leaves from 10 different infected plants were collected for pathogen isolation. The small pieces of discolored leaves were surface sterilized with 75% ethanol for 40s and washed with sterile water for three times. The sterilized leaves were ground with a glass rod with 1mL sterile water, and 100 µL suspensions were spread on nutrient agar medium then incubated at 28oC for 48 hours. Yellow round colonies with undulating edges were showed up on nutrient agar medium 48 hours later. Three isolates were randomly picked up from each of the 10 plates for subsequent analysis. After purification and culture on nutrient agar plate, the 16S rRNA gene of the 30 isolates were amplified with primers 27F and 1492R and the amplicons were sequenced and analyzed by sequence alignment. The sequence alignment results showed that the 16S rRNA nucleotide identity of the 30 isolates were 100%. One typical isolate named ZC5 was selected for subsequent analysis, and the resulting 16S rRNA sequence was deposited at GenBank, NCBI under accession OK092624. The 16S rRNA sequence identity with those of P. psychrotolerans strain K3-2 (KY882083) and M3-1 (KY882120) were 100%, respectively. The phenotypic analysis by Biolog Gen Ⅲ indicated that the bacterial isolate (ZC5) showed highest similarity (98.3%) with strain Pseudomonas oryzihabitans. P. oryzihabitans and P. psychrotolerans have a high degree of homology in the phylogenetic relationship based on the phylogenetic analysis of three concatenated sequences of gyrB, rpoB and rpoD genes (Mulet et al. 2010). The gyrB (ON462356), rpoB (ON462355), rpoD (ON462357) gene of isolate ZC5were also amplified and sequenced by using primers gyrB-For/gyrB-Rev, rpoB-For/rpoB-Rev and rpoD-For/rpoD-Rev (Hauser et al. 2004), respectively. While P. psychrotolerans and P. oryzihabitans form the same clade in phylogenies, strains of P. psychrotolerans do form a unique sub-clade. Isolate ZC5 clustered more closely with the type strain of P. psychrotolerans LMG 21977 in the phylogenetic tree. Therefore, based on the concatenated sequences of three genes (gyrB, rpoB and rpoD), the isolate ZC5 was confirmed as P. psychrotolerans. Based on morphological, Biolog characteristics and phylogenetic analysis, the isolate was identified as P. psychrotolerans. The tobacco plants at fast growing stage were selected for pathogenicity tests. Pathogenicity tests were conducted by injecting 10 µL bacterial suspension (108cfu/mL) of ZC5 into tobacco leaves with a syringe. Sterile water was inoculated into the tobacco leaves in the same way as the control. Six plants were selected for pathogenicity tests each time and five leaves of each tobacco plant were inoculated, and the tests were repeated three times. To simulate disease conditions in the natural environment, the inoculated plants were moved outdoors. The average temperature was 32°C during the day and 20°C at night. To maintain humidity, the tobacco leaves were sprayed with water every two days. Symptoms appeared on the pathogen inoculated leaves seven days after inoculation, whereas the control treatment remained symptomless. The pathogens were reisolated from diseased leaves and identified as P. psychrotolerans based on morphological, molecular and phylogenetic analysis, which fulfilled Koch's postulates. To our knowledge, this is the first report of tobacco bacterial leaf spot caused by P. psychrotolerans.

Challenge of evolving Klebsiella pneumoniae infection in patients on hemodialysis: from the classic strain to the carbapenem-resistant hypervirulent one.

Loss of renal function may render hemodialysis patients more susceptible to infectious diseases, which is the second of all-causes mortality in this population. In addition to infection caused by the classic Klebsiella pneumoniae (cKp), however, hemodialysis staffs are now facing new challenge with growing prevalence of the carbapenem-resistant Kp (CR-Kp) and hypervirulent Kp (hvKp) as they are respectively associated with increased drug-resistance and virulence. We therefore chose to share our recent experience in treating severe infections either caused (cKp, CR-Kp, hvKp) or complicated (CR-hvKp) by these strains in hemodialysis patients. Based upon yet beyond published works, we further came up with the detection of intracranial lesion, novel diagnostic approach using unique biomarkers followed by selection of appropriate antibiotics, management of metastasic abscesses and bracing for the most lethal scenario in the order of cKp, CR-Kp, hvKp and CR-hvKp, respectively. Since reports of complicated hvKp infection in hemodialysis patients were rare, we also discussed in details this clinical entity focusing on its epidemiology, mechanism of increased virulence and involvement of the arteriovenous fistula as insidious source of persistent septicemia. By covering the full spectrum of clinically relevant Kp stains specifically from the viewpoint of nephrology, our work had highlighted the importance of infection control in uremic state and vice versa. As such, it may greatly raise the awareness of dialysis staffs against the challenge of evolving Klebsiella pneumoniae infection in hemodialysis patients and expeditiously reach a higher degree of readiness which was proved to be the key determinant of ultimate survival.

CircSMAD4 alleviates high glucose-induced inflammation, extracellular matrix deposition and apoptosis in mouse glomerulus mesangial cells by relieving miR-377-3p-mediated BMP7 inhibition.

BACKGROUND: Diabetic nephropathy (DN) is a common complication of diabetes mellitus. Accumulating studies suggest that the deregulation of circular RNA (circRNA) is involved in DN pathogenesis. This study aimed to investigate the role of circSMAD4 in DN models. METHODS: Mice were treated with streptozotocin to establish DN models in vivo. Mouse glomerulus mesangial cells (SV40-MES13) were treated with high glucose to establish DN models in vitro. The expression of circSMAD4, miR-377-3p and bone morphogenetic protein 7 (BMP7) mRNA was measured by quantitative real-time PCR (qPCR). The releases of inflammatory factors were examined by ELISA. The protein levels of fibrosis-related markers, apoptosis-related markers and BMP7 were checked by western blot. Cell apoptosis was monitored by flow cytometry assay. The predicted relationship between miR-377-3p and circSMAD4 or BMP7 was validated by dual-luciferase reporter assay or pull-down assay. RESULTS: CircSMAD4 was poorly expressed in DN mice and HG-treated SV40-MES13 cells. HG induced SV40-MES13 cell inflammation, extracellular matrix (ECM) deposition and apoptosis. CircSMAD4 overexpression alleviated, while circSMAD4 knockdown aggravated HG-induced SV40-MES13 cell injuries. MiR-377-3p was targeted by circSMAD4, and miR-377-3p enrichment partly reversed the effects of circSMAD4 overexpression. BMP7 was a target of miR-377-3p, and circSMAD4 regulated BMP7 expression by targeting miR-377-3p. MiR-377-3p overexpression aggravated HG-induced injuries by suppressing BMP7. CONCLUSION: CircSMAD4 alleviates HG-induced SV40-MES13 cell inflammation, ECM deposition and apoptosis by relieving miR-377-3p-mediated inhibition on BMP7 in DN progression.

Sequencing and phylogenetic characterization of a novel RNA virus in Arma chinensis.

The complete genome of a novel virus from Arma chinensis was determined by RNA sequencing and rapid amplification of cDNA ends. This virus has a single-stranded RNA genome of 10,540 nucleotides (nt) excluding the poly(A) tail. Two non-overlapping open reading frames (ORFs) in the sense direction were predicted: one long ORF at the 5' end of the genome (6,219 nt) that encodes a polypeptide of 2,072 amino acids (aa), and one short ORF at the 3' end of the genome (3,033 nt) that encodes a polypeptide of 1,010 aa. Phylogenetic analysis indicated that the virus clusters within a large cluster of currently unidentified picorna-like viruses with a high bootstrap value. We named the virus isolate Arma chinensis picorna-like virus 1 (AcPV-1). The prevalence of AcPV-1 infection in samples of Arma chinensis from the wild was at a low level (5.48%, 8 positives in 146 samples). Keywords: Arma chinensis; genomic characterization; phylogenetic analysis; Arma chinensis picorna-like virus 1; prevalence.

Global patterns in genomic diversity underpinning the evolution of insecticide resistance in the aphid crop pest Myzus persicae.

The aphid Myzus persicae is a destructive agricultural pest that displays an exceptional ability to develop resistance to both natural and synthetic insecticides. To investigate the evolution of resistance in this species we generated a chromosome-scale genome assembly and living panel of >110 fully sequenced globally sampled clonal lines. Our analyses reveal a remarkable diversity of resistance mutations segregating in global populations of M. persicae. We show that the emergence and spread of these mechanisms is influenced by host-plant associations, uncovering the widespread co-option of a host-plant adaptation that also offers resistance against synthetic insecticides. We identify both the repeated evolution of independent resistance mutations at the same locus, and multiple instances of the evolution of novel resistance mechanisms against key insecticides. Our findings provide fundamental insights into the genomic responses of global insect populations to strong selective forces, and hold practical relevance for the control of pests and parasites.

Efficacy of an improved method to screen semiochemicals of insect.

BACKGROUND: A combination of gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS) is typically used to screen active compounds that play a role in the regulation of insect behavior. This method uses two kinds of gas chromatography (GC) equipment and needs to compare compounds between the two chromatograms, and it is tedious and costly. To improve detection efficiency, as well as reduce costs and the rate of missed detection, we designed a system connecting gas chromatography (GC), electroantennography (EAG), and mass spectrometry (MS), with MS used instead of the flame ionization detector (FID) as the GC-EAD detector. To verify the feasibility of the improved method, we compare two methods-GC-EAG-MS and GC-EAD-through a series of experiments. Some researchers made similar improvements, but these were not compared with GC-EAD, and their method needed to be improved in the synchronization and split ratio. Our method has been optimized and improved in these aspects. METHODS: Helicoverpa armigera was the test organism; the improved method and conventional method were used to detect known and unknown compounds, as well as screen out active compounds that could generate responses in H. armigera antennae. RESULTS: Screening known single compounds using the two methods, the active compound benzaldehyde was detected in all seven concentrations of solution. By using the two methods, the five same active compounds of Helicoverpa armigera were detected in high concentration solution of the mixed compounds (100 mg L-1, 50 mg L-1); the four same active compounds were detected at 20 mg L-1 concentration; two identical same compounds were detected in low concentration solution (concentrations of 10 mg L-1 and below). By using the two methods, six identical active compounds of Helicoverpa armigera were detected in unknown compounds. CONCLUSION: The improved method was consistent with the conventional method in terms of accuracy and sensitivity. However, compared with the traditional methods, Gas chromatography-electroantennographic-mass spectrometry (GC-EAG-MS) saved the cost of GC and FID equipment, thereby greatly lowering the experimental cost. In the experiment, GC-EAG-MS combined the two experimental operations of screening active substances by GC-EAD and identifying active substances by GC-MS into one, which not only reduced the experimental steps, but also avoided the false positive caused by the comparison of the two chromatograms, and it greatly reduced the difficulty level of the overall experimental analysis. GC-EAG-MS is more convenient, efficient, economical, and practical, and could confidently replace traditional methods. With further optimization, it could be widely applied in the study of plant and insect chemical ecology.

Sequencing and phylogenetic characterization of a novel Polerovirus from Nicotiana tabacum.

In this study, we reported the complete genome of a novel Polerovirus, named Tobacco yellow virus (TYV), which can be transmitted by Myzus persicae. TYV had a single-stranded RNA genome of 5735 nucleotides in length and contained six putative open reading frames (ORFs). Phylogenetic analysis with whole genome nucleotide sequences and amino acid sequences deduced from the conserved domain of the RNA-dependent RNA polymerase, clustered TYV with Potato leafroll virus from the genus Polerovirus with high bootstrap values. However, TYV clustered with Brassica yellow virus using amino acid sequences deduced from the conserved domain of the coat protein. Taken together with the identities between ORFs in TYV and related ORFs in species from Polerovirus, our results strongly suggested TYV is a novel species of the genus Polerovirus.

First report of Colletotrichum chlorophyti causing peanut anthracnose in China.

Peanut (Arachis hypogaea L.) is an important oil crop - mainly in Shandong and Henan Provinces, in China. In 2018-2019, the occurrence of a black spot disease on the leaves and stems were found in two fields (a total of 10 ha) in Niulingguanzhuang village of Yishui county in Linyi, Shandong Province, China, and 20% to 40% of plants were infected, thereby reducing the amount of marketable product. Natural symptoms were circular, dark brown-to-black lesions (2-6 mm in diameter) and coalescent necrosis on leaves and black necrosis in stems. Six symptomatic leaves and stems collected from six plants of two fields, were used for isolation. Portions of infected tissue were surfaced-sterilized with 0.5% NaClO for two minutes, 70% alcohol for 30 seconds and washed twice with sterile water. The tissues were placed on potato dextrose agar (PDA) at 28℃ for 5 days. Felt or fleece forming colonies about 4 cm in diameter that were circular, flat, and with dark center and white narrow margins were formed. Three isolates (LSJF-4, HSGF, HSJF) were purified and one culture (LSJF-4) was deposited in the China General Microbiological Culture Collection Center (CGMCC, NO. 3.19617). The chlamydospore were dark brown, verruculose, organized in chains and clusters, yellow-brown to black-brown color, and mostly spherical, 10.3 - 18.4 µm × 8.3-11.4 µm (n = 50, av. 12.6 ± 2.1× 10.1 ± 1.1 µm). The conidia were colorless, crescent or sickle-shaped, with one acute end and one blunt end. They were 16.7 - 24.3 µm × 3.6 - 5.5 µm (n= 50, av. 21.2 ± 1.7× 4.5 ± 0.36 µm), with dark brown, straight, septate setae, 40.3 - 86.9 µm × 3.6-5.5 µm (n=50, av. 70.0 ± 19.1 × 4.5 ± 0.7 µm) in size. Setae were straight, dark brown. These morphological characteristics of our isolates were identical to Colletotrichum chlorophyti (Damm et al. 2009). The internal transcribed spacer (ITS, ITS1/ITS4) rDNA of three fungus (LSJF-4, HSGF, HSJF) were amplified using PCR and sequenced, as described by Damm et al (2009). The sequencing results (GenBank Accession No. MK796409, MN756650, MN756651) were submitted to the GenBankand blast analysis showed that it had 99.0% identity with those of Colletotrichum chlorophyti (No. GU227894). Actin (ACT, ACT-512F/ACT-783R), beta-tubulin (Tub2, T1/T2 ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, GDF1/GDR1), chitin synthase (CHS1, CHS-79f/CHS-345R), histone (H3, Cy1H3F/ Cy1H3R) of LSJF-4 (Nos. MN688800, MN688797, MN097811, MN688799 and MN688798, respectively) (Damm et al. 2009) were also sequenced and BLAST. The results showed high identity of all the five sequences to the CGMCC 3.19617 (Nos. GU228286/GU228287 in GAPDH = 99.6%, GU228384/GU22385 in CHS1 = 96.8%, GU228090/GU228091 in H3 = 99.0%, GU227992/GU227993 in ACT = 98.7%, and GU228188/GU228189 in Tub2 = 98.8%). Therefore, isolate CGMCC 3.19617 was identified as C. chlorophyti based on morphological and molecular characteristics. To determine pathogenicity, a conidial suspension containing 106 conidia/mL was sprayed on leaves and stems of six 40-day old peanut seedlings. Three control plants were similarly sprayed with sterile water. All treated plants were kept moist (>85% relative humidity) at 30℃ for 48 h in darkness, and then kept in 60% relative humidity and 28℃ conditions. Two weeks post-inoculation, small, dark, near elliptical lesions appeared on inoculated leaves and stems, which were similar to those naturally infected plants. While controls remained symptomless. C. chlorophyti was reisolated from infected tissues and identified based on the previous methods, fulfilling Koch's postulates. The test was repeated twice. C. chlorophyti has been reported as a pathogen associated with soybean anthracnose isolated from the Netherlands (Damm et al. 2009). C. chlorophyti was first detected on soybean plants imported from Uruguay (Li et al. 2017) in China. To our knowledge, this is the first report of C. chlorophyti causing peanut anthracnose in China. C. chlorophyti may be a new threat to Leguminous plant species, especially peanuts plants. We proposed paying close attention to taking necessary control meatures.