Rice small secreted peptide, OsRALF26, recognized by FERONIA-like receptor 1 induces immunity in rice and Arabidopsis.

Rapid alkalinization factors (RALFs), belonging to a family of small secreted peptides, have been considered as important signaling molecules in diverse biological processes, including immunity. Current studies on RALF-modulated immunity mainly focus on Arabidopsis, but little is reported in crop plants. The rice immune receptor XA21 confers immunity to the bacterial blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo). Here, we pursued functional characterization of rice RALF26 (OsRALF26) up-regulated by Xoo during XA21-mediated immune response. When applied exogenously as a recombinant peptide, OsRALF26 induced a series of immune responses, including pathogenesis-related genes (PRs) induction, reactive oxygen species (ROS) production, and callose deposition in rice and/or Arabidopsis. Transgenic rice and Arabidopsis overexpressing OsRALF26 exhibited significantly enhanced resistance to Xoo and Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), respectively. In yeast two-hybrid, pull-down assays, and co-immunoprecipitation analyses, rice FER-like receptor 1 (OsFLR1) was identified as a receptor of OsRALF26. Transient expression of OsFLR1 in Nicotiana benthamiana leaves displayed significantly increased ROS production and callose deposition after OsRALF26 treatment. Together, we propose that OsRALF26 induced by Xoo in an XA21-dependent manner is perceived by OsFLR1 and may play a novel role in the enforcement of XA21-mediated immunity.

Rice NLR protein XinN1, induced by a pattern recognition receptor XA21, confers enhanced resistance to bacterial blight.

KEY MESSAGE: Rice CC-type NLR XinN1, specifically induced by a PRR XA21, activates defense pathways against Xoo. Plants have evolved two layers of immune systems regulated by two different types of immune receptors, cell surface located pattern recognition receptors (PRRs) and intracellular nucleotide-binding domain leucine-rich repeat-containing receptors (NLRs). Plant PRRs recognize conserved molecular patterns from diverse pathogens, resulting in pattern-triggered immunity (PTI), whereas NLRs are activated by effectors secreted by pathogens into plant cells, inducing effector-triggered immunity (ETI). Rice PRR, XA21, recognizes a tyrosine-sulfated RaxX peptide (required for activation of XA21-mediated immunity X) as a molecular pattern secreted by Xanthomonas oryzae pv. oryzae (Xoo). Here, we identified a rice NLR gene, XinN1, that is specifically induced during the XA21-mediated immune response against Xoo. Transgenic rice plants overexpressing XinN1 displayed increased resistance to infection by Xoo with reduced lesion length and bacterial growth. Overexpression of autoactive mutant of XinN1 (XinN1D543V) also displayed increased resistance to Xoo, accompanied with severe growth retardation and cell death. In rice protoplast system, overexpression of XinN1 or XinN1D543V significantly elevated reactive oxygen species (ROS) production and cytosolic-free calcium (Ca2+) accumulations. In addition, XinN1 overexpression additionally elevated the ROS burst caused by the interaction between XA21 and RaxX-sY and induced the transcription of PTI signaling components, including somatic embryogenesis receptor kinases (OsSERKs) and receptor-like cytoplasmic kinases (OsRLCKs). Our results suggest that XinN1 induced by the PRR XA21 activates defense pathways and provides enhanced resistance to Xoo in rice.

Challenges and solutions in developing an objective and structured clinical examination for complementary and integrative medicine: A mixed-method approach.

This study explores the challenges encountered by developers when creating objective and structured clinical examination (OSCE) modules specifically for Korean medicine (KM). The complexity of developing pattern identification (PI) items, due to the lack of standardized materials and ambiguity in KM, was a primary focus. A mixed-method approach was utilized, including a survey, importance-performance analysis, and focus group interviews. Seven developers participated, creating a total of 21 OSCE modules. The main difficulties identified were in developing PI items, selecting appropriate cases, crafting realistic examination situations, determining scoring criteria, setting up checklists, and writing scenarios. Challenges were categorized into "case," "examination situation," "postexamination notes," "checklist," "scenario," "format," and "PI." The importance-performance analysis revealed improvements in module development capabilities with each iteration. For the future development of efficient OSCE modules, standardization of KM diagnostic methods and PI is essential. The study highlights the need for social and academic efforts, as well as support from the KM education community and schools, to address these challenges and enhance the development process.

Implementation strategy for introducing a clinical skills examination to the Korean Oriental Medicine Licensing Examination: a mixed-method modified Delphi study

PURPOSE: This study investigated the validity of introducing a clinical skills examination (CSE) to the Korean Oriental Medicine Licensing Examination through a mixed-method modified Delphi study METHODS: A 3-round Delphi study was conducted between September and November 2022. The expert panel comprised 21 oriental medicine education experts who were officially recommended by relevant institutions and organizations. The questionnaires included potential content for the CSE and a detailed implementation strategy. Subcommittees were formed to discuss concerns around the introduction of the CSE, which were collected as open-ended questions. In this study, a 66.7% or greater agreement rate was defined as achieving a consensus. RESULTS: The expert panel's evaluation of the proposed clinical presentations and basic clinical skills suggested their priorities. Of the 10 items investigated for building a detailed implementation strategy for the introduction of the CSE to the Korean Oriental Medicine Licensing Examination, a consensus was achieved on 9. However, the agreement rate on the timing of the introduction of the CSE was low. Concerns around 4 clinical topics were discussed in the subcommittees, and potential solutions were proposed. CONCLUSION: This study offers preliminary data and raises some concerns that can be used as a reference while discussing the introduction of the CSE to the Korean Oriental Medicine Licensing Examination.

Oryza-Specific Orphan Protein Triggers Enhanced Resistance to Xanthomonas oryzae pv. oryzae in Rice.

All genomes carry lineage-specific orphan genes lacking homology in their closely related species. Identification and functional study of the orphan genes is fundamentally important for understanding lineage-specific adaptations including acquirement of resistance to pathogens. However, most orphan genes are of unknown function due to the difficulties in studying them using helpful comparative genomics. Here, we present a defense-related Oryza-specific orphan gene, Xio1, specifically induced by the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo) in an immune receptor XA21-dependent manner. Salicylic acid (SA) and ethephon (ET) also induced its expression, but methyl jasmonic acid (MeJA) reduced its basal expression. C-terminal green fluorescent protein (GFP) tagged Xio1 (Xio1-GFP) was visualized in the nucleus and the cytosol after polyethylene glycol (PEG)-mediated transformation in rice protoplasts and Agrobacterium-mediated infiltration in tobacco leaves. Transgenic rice plants overexpressing Xio1-GFP showed significantly enhanced resistance to Xoo with reduced lesion lengths and bacterial growth, in company with constitutive expression of defense-related genes. However, all of the transgenic plants displayed severe growth retardation and premature death. Reactive oxygen species (ROS) was significantly produced in rice protoplasts constitutively expressing Xio1-GFP. Overexpression of Xio1-GFP in non-Oryza plant species, Arabidopsis thaliana, failed to induce growth retardation and enhanced resistance to Pseudomonas syringae pv. tomato (Pst) DC3000. Our results suggest that the defense-related orphan gene Xio1 plays an important role in distinctive mechanisms evolved within the Oryza and provides a new source of Oryza-specific genes for crop-breeding programs.

Synergistic effect of Mahaenggamseok-tang in the treatment of pediatric patients with lower respiratory tract infections: A PRISMA-compliant systematic review and meta-analysis.

BACKGROUND: Infants are often hospitalized because of lower respiratory tract infections, and overuse of antibiotics to treat such infections has led to severe problems. Herbal medicines may be more effective and safer than antibiotics. Mahaenggamseok-tang is a common herbal medicine in Asia, but the evidence for its effectiveness in lower respiratory tract infection treatment is insufficient. This review assesses the efficacy of Mahaenggamseok-tang in treating lower respiratory tract infections. METHODS: The study used Chinese, English, and Korean databases, as well as one Japanese database. All included studies were randomized controlled trials comparing Mahaenggamseok-tang with medication to treat lower respiratory tract infections. Studies using Mahaenggamseok-tang plus Western medicine were also included. Standardized mean difference (SMD), risk ratio (RR) with 95% confidence interval (CI), and risk of bias were analyzed using Review Manager 5.4 software. The GRADEpro website was used to assess the reviews. RESULTS: Seventeen randomized controlled trials with a total of 1993 participants were included in the meta-analysis. All studies compared the Mahaenggamseok-tang plus Western medicine group to the Western medicine only group. Meta-analysis showed that Mahaenggamseok-tang affected total effective rate (risk ratio: 1.20, 95% confidence interval [CI]: 1.10-1.31, P < .001), cough disappearance time (SMD: -1.62, 95% CI: -2.30 to -0.95, P < .001), fever disappearance time (SMD: -2.04, 95% CI: -2.87 to -1.21, P < .001), abnormal lung sound disappearance time (SMD: -1.68, 95% CI: -2.43 to -0.93, P < .001), Creactive protein (SMD: -3.18, 95% CI: -4.36 to -1.99, P < .001), procalcitonin (SMD: -5.04, 95% CI: -9.20 to -0.88, P < .05), tumor necrosis factor-α (SMD: -0.84, 95% CI: -1.46 to -0.23, P < .01), IgE (SMD: -2.69, 95% CI: -2.91 to -2.47, P < .001), and adverse events (risk ratio: 0.44, 95% CI: 0.29-0.68, P < .001), but not interleukin-6 (SMD: -1.59, 95% CI: -3.48 to 0.30, P>.05). DISCUSSION: Mahaenggamseok-tang plus Western medicine is more effective and safer than Western medicine alone for treating lower respiratory tract infections. However, the included randomized controlled trials were not randomized well; therefore, better randomized randomized controlled trials are needed to make significant recommendations.PROSPERO registration number: CRD42020165698.

Population Structure of Stagonosporopsis Species Associated with Cucurbit Gummy Stem Blight in Korea.

Gummy stem blight (GSB), a common and serious disease in cucurbits worldwide, is caused by three genetically distinct species: Stagonosporopsis cucurbitacearum (syn. Didymella bryoniae), S. citrulli, and S. caricae. In Korea, however, the three species of Stagonosporopsis have been barely characterized. In this study, 21 Stagonosporopsis isolates were recovered from watermelon (Citrullus lanatus) and muskmelon (Cucumis melo) leaves and stem showing blight symptoms collected from 43 fields in Korea. Sequence analysis performed with an internal transcribed spacer region was not competent to differentiate the Stagonosporopsis isolates. On the contrary, analysis of ß-tubulin (TUB) genes and three microsatellite markers, Db01, Db05, and Db06, successfully differentiated Stagonosporopsis isolates. Further sequence analysis identified two Stagonosporopsis species, S. citrulli and S. caricae, and one previously unknown species of Stagonosporopsis. Representative isolates from three species caused dark water-soaked lesions on the detached watermelon and muskmelon leaves with no significant differences in the aggressiveness. Our results indicate that the S. citrulli, S. caricae, and unknown Stagonosporopsis sp. are all causal agents of GSB for both watermelon and muskmelon. This is the first report of a new species and the population structure of Stagonosporopsis species causing GSB in Korea.

Effects of acupuncture on postoperative recovery and extubation time: A protocol for systematic review and meta analysis.

BACKGROUND: This systematic review protocol aims to provide evidence of the efficacy and safety of acupuncture on postoperative recovery and extubation time. METHODS: The following 11 electronic databases will be searched from inception: The Cochrane Database of Systematic Reviews, MEDLINE, EMBASE, AMED, CINAHL, 1 Chinese database (CNKI), and 5 Korean databases (OASIS, DBpia, RISS, KISS, NDSL). Only randomized controlled trials of acupuncture treatment for postoperative recovery after surgery will be included for review. The selection of the studies, data extraction, and management will be performed independently by 3 researchers. Methodological quality, including the risk of bias, will be assessed using the Cochrane risk of bias assessment tool. RESULTS AND CONCLUSIONS: Our systematic review will provide evidence of the efficacy of acupuncture on postoperative recovery and extubation time. This evidence will provide useful information to practitioners and patients in the field of surgery and complementary medicine. PROSPERO REGISTRATION NUMBER: 2020 CRD42020168411.

Efficacy of traditional herbal medicine for psychological sequelae in COVID-19 survivors: A protocol for systematic review and meta-analysis.

INTRODUCTION: This study is the protocol for a systematic review to evaluate the efficacy of herbal medicine on COVID-19 survivors with psychological sequelae. Currently, there are many COVID-19 survivors with psychological sequelae as COVID-19 has widely spread all over the world. However, there is no critically appraised evidence of the benefit of herbal medicine for COVID-19 survivors with psychological sequelae. MATERIALS AND METHODS: We will search 11 electronic databases from inception to December, 2022: 4 English databases, MEDLINE, PubMed, Excerpta Medica database (EMBASE), and the Cochrane Central Register of Controlled Trials (CENTRAL); 3 Chinese databases, the Chinese National Knowledge Infrastructure, the Chinese Scientific Journal database, and the Wan Fang database; and 4 Korean databases, the Oriental Medical Advanced Searching Integrated System, the Korean Studies Information Service System, the National Digital Science Links, and the Research Information Sharing Service. We will include randomized controlled trials (RCTs), non-RCTs, and quasi-RCTs for all formations of TRADITIONAL herbal medicine versus conventional drug, placebo, and no treatment for COVID-19 survivors. We will only include the COVID-19 survivors with psychiatric symptoms lasting at least 1 month, regardless of their race, sex, and age. DISCUSSION AND CONCLUSIONS: This systematic review will be published in a peer-reviewed journal. The findings will provide evidence and treatment directions for clinicians. This protocol does not need ethical approval because it will be based on published research. PROSPERO REGISTRATION NUMBER: PROSPERO CRD42020210592.

Efficacy and safety of fermented oyster extract for height of children with short stature: a randomized placebo-controlled trial.

BACKGROUND: Some experimental studies have established the effect of oysters on the promotion of body growth. Yet, there is a lack of human clinical studies. The objective of this study was to evaluate the effect of a fermented oyster (FO) extract on the increase in the height of children with stature in the 25th percentile by age. METHODS: In total, 100 children (6-11 years old) were randomly divided into two (FO or control) groups. For 24 weeks, the subjects in the FO group took the FO extract once daily before sleeping, whereas the control group took placebo extracts, simultaneously. We evaluated the height gain, height velocity (HV), height standard deviation score (SDS), urine deoxypyridinoline (DPD), growth hormone (GH), insulin-like growth factor (IGF-1), and IGF binding protein 3 (IGFBP-3). RESULTS: The height gain and height SDS were significantly higher in the FO group than in the placebo group after 24 weeks (height gain: p < 0.001, height SDS: p < 0.005). The HV was also significantly higher in the FO group than in the placebo group after the 6th and 24th week (p = 0.001, p = 0.004). After 24 weeks, we observed a decrease in GH, IGF, and IGFBP-3 in both groups. However, serum IGFBP-3 level in the FO group reduced less than placebo group. CONCLUSION: FO supplementation may help to increase the height of children, and the effect might be mediated via effects on the IGFBP-3 levels.

Incidence of Alternaria Species Associated with Watermelon Leaf Blight in Korea.

Alternaria leaf blight is one of the most common diseases in watermelon worldwide. In Korea, however, the Alternaria species causing the watermelon leaf blight have not been investigated thoroughly. A total of 16 Alternaria isolates was recovered from diseased watermelon leaves with leaf blight symptoms, which were collected from 14 fields in Korea. Analysis of internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and RNA polymerase II second largest subunit (RPB2) were not competent to differentiate the Alternaria isolates. On the contrary, analysis of amplicon size of the histone H3 (HIS3) gene successfully differentiated the isolates into three Alternaria subgroups, and further sequence analysis of them identified three Alternaria spp. Alternaria tenuissima, A. gaisen, and A. alternata. Representative Alternaria isolates from three species induced dark brown leaf spot lesions on detached watermelon leaves, indicating that A. tenuissima, A. gaisen, and A. alternata are all causal agents of Alternaria leaf blight. Our results indicate that the Alternaria species associated watermelon leaf blight in Korea is more complex than reported previously. This is the first report regarding the population structure of Alternaria species causing watermelon leaf blight in Korea.

Mahaenggamseok-tang, a herbal medicine, for lower respiratory tract infections in pediatric patients: A protocol for systematic review and meta-analysis.

BACKGROUND: The aim of this study is to assess the clinical evidence for or against Mahaenggamseok-tang (MHGT) as a treatment for lower respiratory tract infections (LRTIs) in pediatric patients. METHOD: This systematic review will include randomized clinical trials (RCTs) of MHGT, as a treatment for LRTIs, compared with other therapies such as placebo and western medicine. The search terms will be selected according to the medical subject heading. We will search the following databases for systematic reviews from 2000 to Feb 2020: 5 English databases (The Cochrane Database of Systematic reviews, MEDLINE, Excerpta Medica dataBASE, Allied and Complementary Medicine Database, and Cumulative Index to Nursing and Allied Health Literature), 1 Chinese database, 5 Korean databases (Oriental Medicine Advanced Searching Integrated System, DataBase Periodical Information Academic (DBPIA), Research Information Service System, Korean Studies Information Service System, and National Digital Science Library), and 1 Japanese database (J-Stage). All RCTs of decoctions or alternate forms of MHGT will be included. We will search for all parallel or crossover RCTs without language restrictions. The methodological quality of the RCTs will be assessed using Cochrane risk of bias. Furthermore, the studies will be limited to those performed in children under 16 years of age. RESULTS AND CONCLUSIONS: Our systematic review and meta-analysis will provide evidence for MHGT as a treatment for LRTI. The findings can help practitioners and patients recognize more effective and safer therapeutic methods. PROSPERO REGISTRATION NUMBER: CRD42020165698.

The first report of Hepatozoon species infection in leopard cats (Prionailurus bengalensis) in Korea.

Hepatozoon species infections were detected in 4 of 5 heart samples from leopard cats (Prionailurus bengalensis) in Korea examined by light microscopy and PCR. Microscopically, the schizonts of the Hepatozoon species were observed within the parasitophorous vacuoles of unidentified host cells that were located between myocytes. The schizonts were approximately 31 +/- 4 x 19 +/- 3 microm in size; 3 mature schizonts contained approximately 24, 25, and 35 merozoites. In analyses of the partial 18S rRNA gene sequence, the Hepatozoon species infecting the Korean leopard cats was closely related to Brazilian (99.2-99.8% identical) and Spanish (97.0-98.2% identical) isolates of Hepatozoon felis.