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Abstract Eosinophils are a group of granulocytes well known for their capacity to protect the host from parasites and regulate immune function. Diverse biological roles for eosinophils have been increasingly identified, but the developmental pattern and regulation of the eosinophil lineage remain largely unknown. Herein, we utilize the zebrafish model to analyze eosinophilic cell differentiation, distribution, and regulation. By identifying eslec as an eosinophil lineage-specific marker, we establish a Tg(eslec:eGFP) reporter line, which specifically labeled cells of the eosinophil lineage from early life through adulthood. Spatial-temporal analysis of eslec + cells demonstrates their organ distribution from larval stage to adulthood. By single-cell RNA-Seq analysis, we decipher the eosinophil lineage cells from lineage-committed progenitors to mature eosinophils. Through further genetic analysis, we demonstrate the role of Cebp1 in balancing neutrophil and eosinophil lineages, and a Cebp1-Cebpβ transcriptional axis that regulates the commitment and differentiation of the eosinophil lineage. Cross-species functional comparisons reveals that zebrafish Cebp1 is the functional orthologue of human C/EBPεP27 in suppressing eosinophilopoiesis. Our study characterizes eosinophil development in multiple dimensions including spatial-temporal patterns, expression profiles, and genetic regulators, providing for a better understanding of eosinophilopoiesis.

Abstract Background The American College of Medical Genetics and Genomics (ACMG)/Association for Molecular Pathology (AMP) guidelines recommend using variant enrichment among cases as "strong" evidence for pathogenicity per the PS4 criterion. However, quantitative support for PS4 thresholds from real-world Mendelian case–control cohorts is lacking. Methods To address this gap, we evaluated and established PS4 thresholds using data from the Chinese Deafness Genetics Consortium. A total of 9,050 variants from 13,845 patients with hearing loss (HL) and 6,570 ancestry-matched controls were analyzed. Positive likelihood ratio and local positive likelihood ratio values were calculated to determine the thresholds corresponding to each strength of evidence across three variant subsets. Results In subset 1, consisting of variants present in both cases and controls with an allele frequency (AF) in cases ≥ 0.0005, an odds ratio (OR) ≥ 6 achieved strong evidence, while OR ≥ 3 represented moderate evidence. For subset 2, which encompassed variants present in both cases and controls with a case AF 2.27 or allele count ≥ 3) and the PS4_Moderate threshold (allele count ≥ 6), respectively. Reanalysis applying the adjusted PS4 criteria changed the classification of 15 variants and enabled diagnosis of an additional four patients. Conclusions Our study quantified evidence strength thresholds for variant enrichment in genetic HL cases, highlighting the importance of defining disease/gene-specific thresholds to improve the precision and accuracy of clinical genetic testing.

The time-varying radio frequency interference has strong nonlinear dynamic characteristics, which is difficult to be predicted by linear method effectively, making the anti-interference decision without sufficient information support. To solve this problem, a recurrent neural network for spectrum prediction based on time-frequency correlation features is proposed. A sliding window is used to characterize the two-dimensional correlation of time-frequency series, and the spectrum prediction problem is transformed into a problem similar to spatiotemporal sequence prediction. A gradient bridge structure across time frames is added to reduce the attenuation of the gradient in the long time and multi-level network propagation. The training efficiency and network performance are improved by the loss function with better matching. Simulation and experimental results verify the validity of the network prediction results.

Good's buffers have been widely applied in cell/organ culture over the past half a century as biocompatible pH stabilizers. However, the emergence of severe adverse effects, such as cellular uptake, lysosomal autophagic activation, and visible light-induced cytotoxicity, raises serious questions over its biocompatibility while underlying mechanism was unclear. Here we report that riboflavin (RF, component of cell culture medium) generates 1O2, ·OH, and O2•- under visible light exposure during regular cell manipulation. These short half-life reactive oxygen species (ROS) react with tertiary amine groups of HEPES, producing 106.6 μM of H2O2. Orders of magnitude elevated half-life of ROS in the medium caused severe cytotoxicity and systematic disorder of normal cell functions. We have further designed and validated zwitterionic betaines as the new generation biocompatible organic pH buffers, which is able to completely avoid the adverse effects that found on HEPES and derivate Good's buffers. These findings may also open a new avenue for zwitterionic betaine based materials for biomedical applications.

Short-wall block backfill mining (SBBM) technology can effectively solve the problems of coal resources waste, water resources loss and gangue waste accumulation. However, after the long-term effect of mine water, the internal heavy metal ions in the gangue backfill materials may be leached, which have a certain potential impact on the groundwater environment in the mining area. Therefore, according to the technical characteristics of SBBM technology, comprehensive prevention of water resources loss and water resources pollution during SBBM was studied systematically. First, prevention and control of water resources loss in the mining area. Based on the combination relationship of “coal pillar, backfill body-barrier layer-aquifuge”, The development of water-conducting fractures in overlying strata under SBBM were analysed. The control mechanism of water resources protection under the evolution of overlying strata structure induced during SBBM was revealed. Secondly, prevention and control of water resources pollution in mining areas. The migration model of heavy metal ions in the gob was established. Then, the pollution mechanism of gangue backfill materials to water resources was revealed, the migration law of heavy metal ions in gangue backfill materials was analyzed, the comprehensive prevention and control technology of water resources destruction in mining area was summarized, the comprehensive prevention and control technology of water resources loss-pollution in mining area was summarized, and the design method of filling ratio based on water resources loss-pollution prevention was put forward. The results show that the gangue materials were filled into the gob as the backfill body, the gangue materials would be the permanent bearing body, which bore the load of the overlying strata accompanied with the protective coal pillar among the blocks, which could greatly prevents the collapse of the low-level stratum, restrict the water-conducting fractures to penetrate the barrier layer and protect the integrity of the high-level stratum. In addition, the heavy metal ions in gangue backfill materials were subjected to the coupled action of seepage, concentration, and stress and then driven by water head pressure and gravitational potential energy to migrate along the side of the coal pillar and below the floor stratum, during which mine water served as the carrier. The migration distance of heavy metal ions increased with increasing of the permeability of floor strata, leaching concentration of heavy metal ions/pollution source intensity, depth of floor breaks and water level heights, and decreases with the increase of gangue particle size and surrounding rock stress. Based on this, the comprehensive prevention and control technology of water resources loss-pollution in mining area was summarized, it included mining parameter control technology, backfill parameter control technology, pollution source control technology, transmission route control technology and in-situ water control technology. and the design method of filling ratio based on water resource loss-pollution prevention and control was put forward, which can realize the comprehensive prevention and control of water resource loss-pollution in mining area. This study will provide scientific theoretical basis and guidance for the comprehensive prevention and control of the water resources destruction problems caused by coal mining.

In order to investigate the absorption and retention characteristics of condensed water at different soil layers in the bare desertification area at the bottom of a receding lake in Hoh Xil, Qinghai Province, the characteristics of condensate variation, condensate formation time, condensate volume, and its absorption capacity were investigated in July 2021 using a micro-osmometer. The research area was the artificial water retention layer and bare ground of the exposed sandy bottom formed under the influence of the warming–wetting trend that occurs when water recedes in the salt lake area of the Qinghai-Tibet Plateau, as well as the two conditions of underpass and underseal. According to the results, the time of condensation generation during the observation period in the salt lake area of Hoh Xil begins at about 0:00 and ends at about 10:00. The artificial water retention layer had little influence on the condensation generation time, and the trend of the condensation rate is the same. The unidirectional condensation phenomenon of water vapor in the near-surface air at a depth range of 0–5 cm when an artificial water retention layer is applied is clearly superior to that of other layers; accordingly, its frequency of condensation phenomenon is also the highest. The amount of condensed soil water vapor from the lower part of the ground when an artificial water retention layer is applied at depth ranges of 0–5 cm, 5–10 cm, and 0–10 cm were 3.60, 6.39, and 10.27 times that of the near-surface air, respectively. The total amount of bidirectional condensed water under the condition of the artificial water retention layer at 0–10 cm was the largest. Conversely, the total amount of condensed water from the near-surface air at a depth range of 0–5 cm under the unidirectional condensation condition was the highest, being twice as great as the total amount of condensed water in other layers, which demonstrates the promoting effect that the artificial water retention layer has on the adsorption of water vapor in the near-surface air. Due to the existence of permafrost in the Qinghai-Tibet Plateau, the heat zero-flux plane in the area of the Salt Lake has a migration range of divergence of approximately 30 cm from the surface to the ground. An analogous humidity coefficient characterizes the retention effect of condensed water in certain areas, which serve to demonstrate how the artificial water retention layer distributed at a depth of 0–5 cm, regardless of bidirectional or unidirectional condensation, has the best capacity for adsorption. The results of this study provide a theoretical basis for condensate absorption capacity and vegetation restoration in the bare ecologically degraded areas of the lake bottom. HIGHLIGHTS Explores the exposed sandy area of the Hoh Xil Lake to observe condensation generation patterns and absorption effects.; Investigates the use of artificial retention layers.; Provides a theoretical basis for condensate absorption capacity and vegetation restoration in bare ecologically degraded areas of a lake bottom.; Provides basic data for the rational management, utilization, and ecological restoration of fragile ecosystems.;

Abstract Most of the current bioadhesives cannot perform well on bleeding tissues while postoperative adhesion is a general but serious clinical issue. Here, a three‐layer biodegradable Janus tissue patch (J‐TP) that is able to simultaneously enable efficient closure of bleeding wounds with significantly promoted clotting ability and suppressed postoperative adhesion of tissues is reported. A dry adhesive hydrogel bottom layer of the J‐TP can form rapid (within 15 s) and strong (tensile strength up to 98 kPa) adhesion to bleeding/wet tissues with high bursting pressure (about 312.5 mmHg on a sealed porcine skin) through hydrogen binding and covalent conjugation between the carboxyl & N‐hydroxy succinimide (NHS) groups of hydrogel and the primary amine groups of tissues, while the phosphonic motifs can significantly reduce blood loss (by 81% on a rat bleeding liver model) of bleeding wounds. A thin polylactic acid (PLA) middle layer can improve the tensile strength (by 132%) of the J‐TP in wet conditions while the grafted zwitterionic polymers can effectively prevent postoperative tissue adhesion and inflammatory reaction. This J‐TP may be a promising tissue patch to assist the clinical treatment of injured bleeding tissues with inhibited postoperative adhesion.

Serine/threonine phosphatase calcineurin (CN) is a unique but confounding calcium/calmodulin- mediated enzyme, which is composed of a catalytic subunit A (CNA) and a regulatory subunit B (CNB). We cloned six transcripts for CNA named from NlCNA-X1 to NlCNA-X6, one CNB named NlCNB1 and one CNB homologous gene NlCNBH1 from Nilaparvata lugens. All of them are constitutively transcripted in various tissues and developmental stages. The primary structure of the six isoforms showed obvious differences in the length and composition of the amino acid sequence between the two binding domains of Ca2+/calmodulin (CaM) and CNB. Ca2+-binding EF-hand motifs were found in NlCNB1 and NlCNBH1. The specific gene silencing of NlCNA, NlCNB1 and NlCNBH1 respectively by RNAi resulted in drastical reduction in survival rate, female weight, eclosion rate and fecundity of N. lugens. These results showed that NlCNA, NlCNB1 and NlCNBH1 were required for N. lugens growth and reproduction. The negative effects of NlCNB1 silence on nymph mortality (97%), molting malformation (90%) and female sterile (50%) were more serious than those of NlCNA or NlCNBH1. qRT-PCR and enzyme-linked immunosorbent assay (ELISA) analyses indicated that the nymphs with silenced NlCNA, NlCNB1 or NlCNBH1 showed impaired hormone and energy metabolism. In nymphs, the contents of 20-hydroxyecdysone (20E) after NlCNB1 RNAi and phenoloxidase after NlCNA RNAi were particularly decreased. These results suggested that NlCNA is involved in immunity of N. lugens by regulation of phenoloxidase, while NlCNB1 may control the growth and development of N. lugens by 20E signaling pathway in addition to interact with CNA. Injection of 70 ng/μL dsNlCNB1 resulted in 77.0% down regulation of NlCNB1, and the nymph mortality was up to 57.9% at 10 d after injection. Therefore, NlCNB1 could be a potential candidate target used for strategy design in control of N. lugens. Our results revealed the importance of CN in the regulation of the growth and development of N. lugens, which provided a basis for further study of the molecular mechanism of CN.

This study examines Malay language teachers' perceptions of using Google tools in their online teaching in primary schools. It uses a basic interpretative research methodology to investigate the perspectives of six primary school Malay language teachers from Terengganu, Malaysia. Interviews with semi-structured questions were used to collect data and analyzed thematically. All the themes and sub-themes were organized using NVivo 12 software. The data show that the predominant topic in teachers' thoughts on the importance of Google tools was the reasons for their absence of use. There were two essential sub-themes: a lack of skills and proficiency and a need for more awareness of how to use Google tools. Furthermore, this study highlighted personal support of module development as a secondary theme which included two sub-themes: agreement with the need for modules and the importance of providing them. This study also underlined the importance of module preparation which was reinforced by two sub-themes: facilitating teachers' implementation of online teaching and serving as a significant reference for teachers. This study shows that effective online education requires competency, skills and knowledge of Google Tools. It suggests creating a Google Tools module to guide and serve as a resource for Malay language teachers.

Mastering creative thinking skills (CTS) is crucial for enhancing students' science conceptual understanding. However, conventional teaching prioritizes linear thinking over the three sub-CTS essential to science learning: associative, visual, and divergent thinking. Weaknesses in these three sub-CTS cause students to have misconceptions, difficulty in creating mental images, and an inability to generate ideas on abstract scientific concepts. The aim of this study is to evaluate the effectiveness of the Digital Storytelling Science Creative (DSSC) module in enhancing these three sub-CTS, based on the topic of acids and bases. The ASSURE instructional design model guided the DSSC module design and development process. This quasi-experimental design study involved 63 eighth-grade students. Analysis of two-way MANCOVA using the data collected through the administered Creative Thinking Skills Test revealed that the DSSC module group performed significantly better than the control group in divergent thinking. However, the DSSC module did not significantly affect students' associative and visual thinking or gender.

The percentage of international faculty members at Korean universities peaked in 2013 and has been decreasing since, suggesting that Korean universities have a problem with international faculty member retention. This study investigates whether international faculty members' intention to leave is associated with their degree of acculturation. Results are based on analyses of data from 325 international faculty members with doctorates working at Korean universities nationwide. Regression analyses reveal that acculturation, work-related conditions (salary, workload, workplace climate) and mobility-related motivational factors (job market in home country, interest in Korea) explain international faculty members' intention to leave. A 'cultural advantage' (ethnically Korean or living with a Korean spouse) is not a significant predictor of intention to leave. Suggestions to support international faculty members, and to help bring about synergistic organizational change are ventured in the hopes of bolstering retention at Korean universities going forward.

This study aims to obtain an e-book based on the Realistic Mathematics Education (PMR) approach that is valid, practical, and effective so that it can improve students' mathematical communication skills on the Two-Variable Linear Equation System (SPLDV) material. This type of research is Research and Development (R&D) using the ADDIE model which consists of five stages, namely analyze, design, development, implementation, and evaluation. The research instruments used were questionnaire and instrument validation sheets, lesson plans validation sheets, e-book validation sheets, mathematical communication skills tests, and student and teacher response questionnaires to e-books. Based on the research, the results of the RPP validation were obtained with a percentage of 88% (very valid), e-book validation with a percentage of 82% for material, and 90% for media with a very valid category. The results of student responses with the percentage of the practicality of 86% (very practical) and the teacher's response questionnaire with the percentage of the practicality of 86% (very practical). The results obtained by meeting the effective criteria with a classical student learning completeness score of 96%, more than 65% of students achieving 75% of the learning objectives for each indicator, and the learning time is the same as ordinary learning in field trials and student responses are good to the e-book provided. developed. Through the Gain test, it is seen that students' mathematical communication skills using e-books based on the PMR approach have increased by 0.7, meaning that they are in the high category. Keywords: e-book, realistic mathematics education, mathematical communication.

Huiming Jiang,1,* Haibin Chen,2,* Pei Wan,1 Meng Liang,3 Nanhui Chen1 1Department of Urology, Meizhou People’s Hospital, Guangdong Provincial Key Laboratory of Precision Medicine and Clinical Translational Research of Hakka Population, Meizhou, People’s Republic of China; 2Department of Histology and Embryology, Shantou University Medical College, Shantou, People’s Republic of China; 3Gannan Medical University, Ganzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Huiming Jiang; Nanhui Chen Tel +86-13560990839Email jianghm0228@163.com; chennanhuiht@163.comBackground: Clear cell renal cell carcinoma (ccRCC) is the most common and aggressive type of primary kidney cancer worldwide. Transmembrane protein 45A (TMEM45A) has been reported to be closely associated with the progression of several cancers. However, the role of TMEM45A in ccRCC remains unclear. Our study intended to explore the potential role of TMEM45A in ccRCC.Methods: Data on the expression of TMEM45A were obtained from multiple databases, including UCSC, GEPIA2, Oncomine and TIMER. Real-world samples of ccRCC and paired normal renal tissues were used to confirm the information obtained from the databases. In addition, the prognostic value of TMEM45A was evaluated. Loss-of-function assays were performed using TMEM45A-targeting lentivirus to evaluate the biological role of TMEM45A in renal cancer cells. Gene set enrichment analysis (GSEA) was performed to investigate the potential molecular mechanisms.Results: TMEM45A was significantly overexpressed in patients with ccRCC and correlated with poor overall survival and disease-free survival. In addition, the expression of TMEM45A was closely associated with various clinicopathological parameters such as histological grade and TNM stage. Knockdown of TMEM45A inhibited the proliferation and migration and promoted the apoptosis of ccRCC cells in vitro. The results of the GSEA suggested that TMEM45A was potentially involved in the promotion of epithelial–mesenchymal transition (EMT) and inflammatory response in ccRCC.Conclusion: TMEM45A was overexpressed and associated with poor survival and acted as a tumour promoter in ccRCC; therefore, might be a potential prognostic marker and therapeutic target.Keywords: biomarker, clear cell renal cell carcinoma, prognosis, TMEM45A

Objective To investigate the effects of novel turtle shell decoction (NTSD) on the cycle, apoptosis, connective tissue growth factor/integrin alpha 5 (CTGF/Itgα5) pathway and the expression of Collagen type I (Col1a1) in rat hepatic stellate cell line T6 (HSCT6). Methods HSCT6 cells was treated with NTSD containing crude drug 0.71 g/mL as experimental group, treated with turtle shell pills (TSP) at the same dose as positive control, and cultured routinely as normal control. Cell cycle and apoptosis of HSCT6 cells were tested by flow cytometry (FCM). Real time-qPCR, Western blotting and immunofluorescence assay were employed to detect the expression changes of CTGF, Itgα5 and Col1a1 genes and their coding proteins in each group of HSCT6, respectively. Results FCM showed that NTSD treatment blocked the cells at S stage, and induced cell apoptosis. Real time-qPCR, Western blotting and immunofluorescence assay revealed that NTSD can significantly down-regulate the expression of CTGF, Itgα5 and Col1a1 in mRNA and protein levels in HSC-T6 cells (P < 0.01). Conclusion NTSD treatment results in cell arrest at S phage through down-regulating CTGF/Itgα5, induction of cell apoptosis, decrease of Col1a1 expression, and then inhibition of cell growth in HSCT6 cells.

The paper establishes a related differential equation model about changes in financial interest rates. It uses information related to liquidity to feedback the law and stability of differential equations in interest rate changes. The article applies stochastic processes and partial differential equations to complex financial networks to confirm node yields in financial market networks. It confirms the existence of interest rate stickiness in Chinese financial markets. The advantage of this interest rate model is that when the external economic environment changes, the state of interest rates will also change accordingly.

Jinyi Min,1,&ast; Peng Wan,2,&ast; Guiwei Liu,1 Min Yu,2 Lei Su3 1Department of Critical Care Medicine, The People’s Hospital, Dangyang City, Hubei, 444100, People’s Republic of China; 2Department of Critical Care Medicine, The People’s Hospital of China Three Gorges University, Yichang City, Hubei, 443000, People’s Republic of China; 3Department of Critical Care Medicine, Guangzhou General Hospital of Guangzhou Military Command, The Military Key Laboratory of Trauma Care in Hot Zone and Tissue Repair in PLA, Guangzhou, 510000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Peng WanDepartment of Critical Care Medicine, The People’s Hospital of China Three Gorges University, 4th Street Hudi, Xiling District, Yichang City, Hubei, 443000, People’s Republic of ChinaTel + 86717-6287551Fax + 86717-6221636Email wanpeng0a0@163.comPurpose: Data regarding the incidence of a coagulable state following heat stroke as assessed by Sonoclot signature analysis are limited. Our purpose was to appraise coagulopathy using a dynamic test capable of analyzing the entire coagulation cascade and to characterize coagulation in patients with heat stroke prior to transfusion.Materials and Methods: The data of 106 patients were collected prospectively from the Critical Care Center of the General Hospital of Guangzhou Military Command. Coagulable state was defined as normal. Both hyper- and hypo-coagulable states were defined as coagulation defects. Hypercoagulability was defined as an activated clotting time (ACT) ≦195s and a clot rate (CR) > 23, and hypocoagulability was defined as an ACT ≧119s and a CR < 7. The Sonoclot signature t examination was performed at the time of admission. Conventional tests, such as the prothrombin time (PT) and activated partial thromboplastin time (aPTT), were compared with Sonoclot monitoring to identify coagulation defects.Results: The average age of the 106 patients was 23.2± 2.5 years. There were 102 males (96.3%) and 4 females (3.7%). Thirty-four patients (32.1%) were hypercoagulable and 44 patients (41.5%) were hypocoagulable at the time of admission; 28 patients (26.4%) had no evidence of a coagulopathy. Patients with hypocoagulability, unlike patients with hypercoagulability, had a higher sequential Organ Failure Assessment score, indicating a more severe multiple organ dysfunction score. Mortality was 5.9% in patients with hypercoagulability compared with 3.5% in patients with normal coagulation, and 18.1% in patients with a hypocoagulable state (P < 0.05). ACT was a predictor of mortality, while the CR and platelet function did not show statistical significance.Conclusion: This study determined the clinical outcomes and prognostic value of coagulability in patients with heat stroke, as defined by Sonoclot signature analysis at the time of admission.Keywords: heat stroke, coagulopathy, prognosis

Studies found that the mastery of Level-4 Science Process Skills (L4SPS) among secondary school students is still relatively weak. Thus, the purpose of this research was to determine the effects of 5E Inquiry-based Learning and Group Investigation (I5E-GI) model on the five constructs of L4SPS, namely Identifying Manipulated, Responding, and Constant Variables, Forming Hypotheses, Making Observations, Making Inferences, and Defining Operationally. A teaching and learning (TL) module was developed to guide teachers in implementing the I5E-GI method for the five L4SPS constructs. The L4SPS Test Instrument was constructed to measure the level of L4SPS mastery at the end of intervention. The quasi-experimental research design was conducted on 180 Form Four students taking Core Science subjects. A total of three groups were assigned, namely i) 5E Inquiry-based Learning and Group Investigation method (I5E-GI, n = 60), ii) 5E Inquiry-based Learning method (5E, n= 60), and iii) Traditional Learning (TR, n = 60) in an urban secondary school in Sarawak, Malaysia. Data were analyzed using the inferential statistical tests of MANOVA, MANCOVA, ANCOVA, and effect size. The results showed that there was a statistically significant effect across the three groups of TL methods. There is a statistically significant effect of the I5E-GI method compared to the 5E and TR methods on the five L4SPS constructs. As for the effect size, the I5E-GI method provides a more significant effect size than the 5E and TR learning methods. Overall, the findings prove that the I5E-GI method has positive implications for the mastery of L4SPS among Form Four students.

Purpose: Although there exists a substantial body of research discussing the efficient implementation of classroom-based assessment (CBA), there is a dearth of knowledge regarding its application within the context of English as a Second Language (ESL) education in Malaysia. The current study aimed to investigate the ESL teachers' practices in implementing CBA, with a specific emphasis on the alignment of their assessment procedures with the policy requirements. Methodology: Hence, this research utilizes a qualitative approach, employing data triangulation through semi-structured interviews, classroom observations, and document analysis. Four ESL teachers from two public secondary schools in the northeast district of the Malaysian city of Pulau Pinang were selected to participate in the study. The four ESL teachers taught the same grade (Form 2) at the secondary level. The data collected were analyzed through thematic analysis procedures. Findings: This research discovered that teachers demonstrate commendable practices in planning the CBA. They exhibited discernment by formulating instructional objectives in line with the curriculum document. However, a misalignment was observed in teachers' practices during the implementation phase. Despite employing various assessment methods, teachers predominantly leaned towards formal assessments, with less emphasis on student-centred approaches. Challenges surfaced when explicit learning objectives were not effectively communicated, and the use of standardized rubrics, was not tailored to students' diverse capabilities. At the reporting phase, concerns were raised about an excessive focus on administrative data recording. This study brought to light that teachers relied on the accumulation of students' grades and marks to determine the mastery levels, thus corroborating with the examoriented practices. Significance: This study holds significant value for the field of ESL education in Malaysia by shedding light on the actual implementation of CBA and its alignment with policy directives. By providing information on how policies are implemented at the local level, contributes to the improvement of assessment policies that can be tailored to classroom realities.

The purpose of this study was to explore the influence of educational choice on the intention to enter the examination, and the mediating role of career decision-making self-efficacy. A total of 1391 freshmen from six higher vocational colleges in Guangdong Province of China were investigated with Education Choices Scale, Career Decision-Making Self-Efficacy Scale and Willingness to Apply Scale. The results showed that educational choice and career decision-making self-efficacy had significant effects on the willingness to apply. Career decision-making self-efficacy plays a mediating role in the significant positive impact of educational choice on the willingness to apply. Therefore, China's educational institutions and policy makers should focus on cultivating vocational students' career decision-making self-efficacy and improving their confidence and ability in career decision-making. To help students better understand their interests, values, and career goals, and to help students better understand the career fields they face professionally.

The authors contend that children benefit from play as a form of intervention and as a means of fostering their cognitive, social, and physical growth. They review several standardized instruments developed over the last fifty years to assess this benefit of play on child development. They identify twenty-one such play measures, the majority of them applicable to children under twelve. Some of the measurements can be conducted by children, but most are proxy rated and employ some form of observation. Over half require no or minimal training. Others can be completed in less than thirty minutes. More evaluations exist for the earlier instruments, but these measurements lacked psychometric evidence to back them up. The authors note they based their selections on the distinct characteristics and features of the measures, and they offer their conclusions to assist users in choosing the best systematic instruments for their specific needs.

Excessive inflammation causes chronic diseases and tissue damage. Although there has been drug treatment, its side effects are relatively large. Searching for effective anti-inflammatory drugs from natural products has become the focus of attention. First isolated from Trichoderma longibraciatum, trichodimerol is a natural product with TNF inhibition. In this study, lipopolysaccharide (LPS)-induced RAW264.7 macrophages were used as a model to investigate the anti-inflammatory activity of trichodimerol. The results of nitric oxide (NO) detection, enzyme-linked immunosorbent assay (ELISA), and reactive oxygen species (ROS) showed that trichodimerol could reduce the production of NO, ROS, and the proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α. Western blotting results showed that trichodimerol could inhibit the production of inflammatory mediators such as cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) and the protein expression of nuclear transcription factor-kappaB (NF-κB), p-IKK, p-IκB, Toll-like receptor 4 (TLR4), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase)-1, and ASC, which indicated that trichodimerol may inhibit inflammation through the NF-κB and NLRP3 pathways. At the same time, molecular docking showed that trichodimerol can directly combine with the TLR4-MD2 complex. Hence, trichodimerol inhibits inflammation by obstructing the interaction between LPS and the TLR4-MD2 heterodimer and suppressing the downstream NF-κB and NLRP3 pathways.

This study explores advertising students' recognition and comprehension of in-feed native advertising on Instagram, a leading platform for social media native advertising. Data were collected from 303 U.S. college advertising students. It examines the impact of two key aspects of Instagram in-feed native advertising--an additional label ("Shop Now") and ad-context congruency--through both quantitative and qualitative data. Findings reveal a significant deficit in native ad recognition among advertising students. Qualitative analysis identified four themes constituting Instagram in-feed advertising for advertising students: professional photo quality, branded products, persuasive elements, and excessive hashtags. Quantitative results indicate that high ad-context congruency led to higher perceptions of nativeness and more positive emotions, but did not significantly influence ad attitudes or negative emotions. The study underscores practical and theoretical implications for advertising educators and scholars, emphasizing the need for enhanced digital advertising literacy and discussing broader implications for advertising pedagogy and practices.

Postpartum depression is a significant public health issue that occurs within the first 12 weeks after childbirth. It is more prevalent among teenage mothers compared to adults. However, the findings of the existing interventional studies cannot be readily applied to teenage mothers due to their unique psychosocial concerns. Furthermore, these findings have shown inconsistencies regarding the benefit of psychological and psychosocial interventions in reducing the incidence of postpartum depression. The current review is aimed at investigating the effectiveness of psychosocial interventions in preventing postpartum depression, specifically among teenage mothers. The preferred reporting items for systematic reviews and meta-analysis manuals were utilized to identify and select relevant articles for this review. The articles were retrieved using population, intervention, control, and outcome models. The quality of each article was assessed using the Cochrane risk of bias tool. Statistical analysis was conducted using STATA version 17. The effect size of the intervention was estimated using the standard mean difference in depression scores between the intervention and control groups. Heterogeneity among the studies was assessed using the I[superscript 2] statistic and Q statistic, while publication bias was evaluated through funnel plot asymmetry and Egger's test. A total of nine eligible articles were included. While psychosocial interventions have been demonstrated to decrease the incidence of postpartum depression compared to usual maternal health care, it is worth noting that the mean difference in depression scores was significant in only three of the included studies. The meta-analysis revealed that psychosocial interventions were effective at preventing postpartum depression, with a pooled effect size of -0.5 (95% CI: -0.95, -0.06) during the final postpartum depression assessment. The heterogeneity was substantial, with an I[superscript 2] value of 82.3%. Although publication bias was not observed, small studies had a significant effect on the pooled effect size. The findings of this review suggest that psychosocial interventions can effectively prevent PPD, particularly within the first 3 months of the postpartum period. This review highlights the scarcity of interventional studies in low-income countries, indicating the need for further research in diverse communities.

This study investigated the impact of problematization-oriented scaffolding and structuring-oriented scaffolding, incorporated within instructional videos, on students' computational thinking and their performance in programming education. We recruited 86 participants from three senior classes at a high school. Each of the three classes was assigned to one of the experimental conditions: the problematization-oriented scaffolding group, the structuring-oriented scaffolding group, and the control group, which received no scaffolding. Pre- and post-tests were conducted to assess students' computational thinking and programming performance during a three-week period. We observed significant improvements in the computational thinking skills of the problematization-oriented scaffolding group, particularly in the algorithmic thinking and cooperativity subdimensions, when compared to the control group. Structuring-oriented scaffoldings in videos also helped enhance students' computational thinking, especially in the algorithmic thinking subdimension. In contrast, students in the control group primarily improved their problem-solving skills, although the difference across the three groups was not significant. Furthermore, this study revealed that the use of learning scaffolds, whether problematization-oriented or structured, significantly contributes to students' learning achievements in comparison to the control group. These findings emphasize the importance of selecting the appropriate scaffolding approach to enhance specific dimensions of computational thinking.

Online and computer-assisted learning have become widespread in the rapidly evolving education landscape. However, these learning modalities uniquely challenge academic integrity, escalating the potential for academic cheating. This systematic review used thematic and narrative syntheses to examine the relationships and the effects of self-doubt and self-regulation on academic cheating in online and computer-assisted learning environments. It involved a sample of 93 peer-reviewed empirical studies published between 1998 and 2023 across five electronic databases adhering to the principles of the PRISMA framework. Findings reveal that different emotional, motivational, and cognitive factors act as primary mediators, while the individual, situational, and environmental factors were significant moderators. These findings underscore the context-dependent and inconsistent effects of self-doubt and self-regulation on academic cheating. Notably, while self-doubt and self-regulation exert opposing influences on academic cheating, other factors, such as gender, culture, performance, feedback, peer pressure, and proctoring, significantly modify these effects. Hence, the relationship between self-doubt, self-regulation, and academic cheating in online and computer-aided learning is thus intricate and dynamic, depending upon various individual, situational, and contextual elements that shape students' motivation, emotions, and cognition. Therefore, this study contributes to the broader discourse on online and computer-aided learning by offering strategies to prevent and reduce academic cheating. Recommendations include promoting self-regulation, reducing self-doubt, focusing on specific mediators and moderators, and utilizing different resources and techniques to measure and identify academic cheating. The results underline the importance of a concerted, multi-faceted approach to upholding academic integrity in the era of digital learning.

Dental anatomy education for dental technology students should be developed in alignment with digital dental laboratory practices. We hypothesized that a virtually assisted sketching-based dental anatomy teaching module could improve students' acquisition of skills essential for digital restoration design. The second-year dental technology curriculum included a novel virtual technology-assisted sketching-based module for dental anatomy education. Pre- and post-course assessments evaluated students' skill sets and knowledge bases. Computer-aided design (CAD) scores were analyzed after one year to assess how the skills students developed through this module impacted their subsequent CAD performance. Participants who undertook the dental sketching-based teaching module demonstrated significantly improved theoretical knowledge of dental anatomy, dental aesthetic perception, and spatial reasoning skills. A partial least squares structural equation model indicated that the positive effects of this module on subsequent CAD performance were indirectly mediated by dental aesthetic perception, spatial reasoning, and practice time. A virtually assisted sketching-based dental anatomy teaching module significantly improved students' acquisition of skills and knowledge and positively mediated dental technology students' CAD performance.

Parent education programs are important avenues through which parents of children with developmental delays can learn new skills to help them with child-rearing challenges. However, those in economic hardship often face problems with applying these skills at home where the child's learning and/or behavioral problems commonly occur. In this exploratory qualitative study, we adopted a positive deviance (PD) lens to explore strategies six economically disadvantaged parents used to support their children who were attending special and mainstream preschools concurrently. The PD perspective was first used in public health research with low-resource communities where a few individuals have been observed to adopt uncommon practices and behaviors that enable them to find better ways to prevent social or health problems than their neighbors facing similar constraints and risks. We identified adaptive parental behaviors, thoughts and feelings that helped them navigate daily hassles faced with "dual schooling" their children in Singapore. The study highlights important implications in the development and refinement of future parent training programs.

In recent years, digital computing in memory (CIM) has been an efficient and high-performance solution in artificial intelligence (AI) edge inference. Nevertheless, digital CIM based on non-volatile memory (NVM) is less discussed for the sophisticated intrinsic physical and electrical behavior of non-volatile devices. In this paper, we propose a fully digital non-volatile CIM (DNV-CIM) macro with compressed coding look-up table (LUT) multiplier (CCLUTM) using the 40 nm technology, which is highly compatible with the standard commodity NOR Flash memory. We also provide a continuous accumulation scheme for machine learning applications. When applied to a modified ResNet18 network trained under the CIFAR-10 dataset, the simulations indicate that the proposed CCLUTM-based DNV-CIM can achieve a peak energy efficiency of 75.18 TOPS/W with 4-bit multiplication and accumulation (MAC) operations.

Most studies on gamified learning have neglected gamification depth, which has motivated the current study to identify the impact of gamified depth on students' intrinsic motivation and performance levels. A quasi-experimental approach was employed, which involved a total of 117 undergraduate students divided into control (n = 57) and experimental (n = 60) groups. The students were exposed to two gamified platforms, namely Kahoot! for shallow gamification and Classcraft for deep gamification, over four weeks. The Intrinsic Motivation Inventory (IMI) served to measure changes that occurred in students' intrinsic motivation levels and the achievement test in the final lesson. Paired t-tests, an independent t-test and regression analysis were conducted to test the study hypothesis. Deep gamification significantly increased learners' intrinsic motivation level (+0.1845), while shallow gamification decreased it (-0.6439). Contrarily, students in the shallow gamified learning environment achieved a higher performance with a mean value of 73.42 compared to the counterparts in a deep gamified environment (50.83). Students' language mastery, interest, attitude and familiarity with technology were postulated to impact the study outcomes. Distinct subjects, age groups and a suitable experiment period are recommended for future studies to develop a proper gamified depth framework in the educational sector.

Nasopharyngeal carcinoma is a diverse cancer characterised by a heterogeneous microenvironment. Here, the authors use single cell sequencing to analyse the tumour microenvironment in 10 nasopharyngeal carcinoma tumours and identify different cell types including immune-suppressive T regulatory, tolerogenic dendritic, and exhausted CD8 T cells.

Abstract Background As core units of organ tissues, cells of various types play their harmonious rhythms to maintain the homeostasis of the human body. It is essential to identify the characteristics of cells in human organs and their regulatory networks for understanding the biological mechanisms related to health and disease. However, a systematic and comprehensive single-cell transcriptional profile across multiple organs of a normal human adult is missing. Results We perform single-cell transcriptomes of 84,363 cells derived from 15 tissue organs of one adult donor and generate an adult human cell atlas. The adult human cell atlas depicts 252 subtypes of cells, including major cell types such as T, B, myeloid, epithelial, and stromal cells, as well as novel COCH + fibroblasts and FibSmo cells, each of which is distinguished by multiple marker genes and transcriptional profiles. These collectively contribute to the heterogeneity of major human organs. Moreover, T cell and B cell receptor repertoire comparisons and trajectory analyses reveal direct clonal sharing of T and B cells with various developmental states among different tissues. Furthermore, novel cell markers, transcription factors, and ligand-receptor pairs are identified with potential functional regulations in maintaining the homeostasis of human cells among tissues. Conclusions The adult human cell atlas reveals the inter- and intra-organ heterogeneity of cell characteristics and provides a useful resource in uncovering key events during the development of human diseases in the context of the heterogeneity of cells and organs.

Background: Excessive inflammation results in severe tissue damage as well as serious acute or chronic disorders, and extensive research has focused on finding new anti-inflammatory hit compounds with safety and efficacy profiles from natural products. As promising therapeutic entities for the treatment of inflammation-related diseases, fusaproliferin and its analogs have attracted great interest. However, the underlying anti-inflammatory mechanism is still poorly understood and deserves to be further investigated.Methods: For the estimation of the anti-inflammatory activity of fusaproliferin (1) and its analogs (2-4)in vitro and in vivo, lipopolysaccharide (LPS)-induced RAW264.7 macrophages and zebrafish embryos were employed. Then, transcriptome analysis was applied to guide subsequent western blot analysis of critical proteins in related signaling pathways. Surface plasmon resonance assays (SPR) combined with molecular docking analyses were finally applied to evaluate the affinity interactions between 1-4 and TLR4 and provide a possible interpretation of the downregulation of related signaling pathways.Results: 1-4 significantly attenuated the production of inflammatory messengers, including nitric oxide (NO), reactive oxygen species (ROS), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), as well as nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in LPS-induced RAW264.7 macrophages. Transcriptome analyses based on RNA-seq indicated the ability of compound 1 to reverse LPS stimulation and the nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPKs) signaling pathways contribute to the anti-inflammatory process. Experimental verification at the protein level revealed that 1 can inhibit the activation of inhibitor of NF-κB kinase (IKK), degradation of inhibitor of NF-κB (IκB), and phosphorylation of NF-κB and reduce nuclear translocation of NF-κB. 1 also decreased the phosphorylation of MAPKs, including p38, extracellular regulated protein kinases (ERK), and c-Jun N-terminal kinase (JNK). SPR assays and molecular docking results indicated that 1-4 exhibited affinity for the TLR4 protein with KD values of 23.5–29.3 μM.Conclusion: Fusaproliferin and its analogs can be hit compounds for the treatment of inflammation-associated diseases.

Objective To study the relation of methotrexate (MTX) dose with blood concentration of MTX polyglutamates (MTX-PGs) and clinical efficacy and explore the optimal initial dose of MTX for treatment of rheumatoid arthritis (RA). Methods We conducted a retrospective analysis of 165 RA patients receiving treatment in our hospital with MTX at the weekly oral dose of 10(n=67), 12.5(n=66) or 15 mg (n=32). The 28 joint disease activity scores (DAS28) were calculated before and after 8 weeks of MTX treatment. According to the European Union Anti-rheumatic Union (EULAR) criteria for efficacy evaluation, the clinical efficacy of the treatment was categorized into favorable responses (including significant and moderate responses) and nonresponse. High-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) assay was used to determine the blood concentration of MTX-PGs of the patients, and relations of MTX dose with the clinical efficacy, DAS28 after treatment and blood MTX-PGs concentration was analyzed. Results The patients taking MTX at the weekly dose of 10, 12.5 and 15 mg showed total response rates of 37.31%, 57.58%, and 56.25%, respectively, which was significantly lower in 10 mg group than in 12.5 mg group (P < 0.05). The median DAS28 after treatment was 3.92 (2.13-5.15), 2.80 (1.81-4.22), and 2.96 (2.26-3.67) in the 10, 12.5, and 15 mg groups, respectively, significantly higher in the 10 mg group than in 12.5 mg group (P < 0.05). The median blood concentrations of MTX-PGs in the 10, 12.5, and 15 mg groups were 92.0 (88.0-96.0), 112.0 (103.0-115.0), and 127.5 ng/mL (102.0-153.8 ng/mL), respectively, showing significant differences among the 3 dose groups (P < 0.05). An increased MTX dose was associated with a higher MTX-PGs concentration in these patients. In 10 mg group, the mean blood concentration of MTX-PGs was 92.5±7.0 ng/mL, significantly lower than the documented concentration (92.5±7.0 ng/mL) required to produce a clinical response (P < 0.05). Conclusion MTX at the weekly oral dose of 12.5 mg can be the optimal initial dose for treatment of local RA patients in Chongqing.

Quan Zhao,1,* Linhai Li,1,* Yu Zhu,1 Dezhi Hou,1 Yuejin Li,1 Xiaodong Guo,1 Yongzhi Wang,1 Opeyemi Joshua Olatunji,2 Ping Wan,3 Kunmei Gong1 1Department of General Surgery, First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), Kunming, Yunnan 650032, People’s Republic of China; 2Faculty of Thai Traditional Medicine, Prince of Songkla University, Hat Yai 90112, Thailand; 3Department of Digestive Internal Medicine, First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), Kunming, Yunnan 650032, People’s Republic of China*These authors contributed equally to this workCorrespondence: Kunmei GongDepartment of General Surgery, First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), Kunming, Yunnan 650032, People’s Republic of ChinaEmail gkmmed@sina.comPing WanDepartment of Digestive Internal Medicine, First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), Kunming, Yunnan 650032, People’s Republic of ChinaEmail kmwp_66@sina.comBackground: Obesity is characterized by excessive body fat, insulin resistance and dyslipidemia, which increases the chances of developing chronic diseases like type 2 diabetes, cardiovascular diseases, hypertension, nonalcoholic fatty liver diseases, some types of cancers and neurodegenerative diseases. Kukoamine B (Kuk B) is a spermine alkaloid obtained from Lycium chinense, and it has been shown to possess antidiabetic, antioxidant and anti-inflammatory properties. In this study, we evaluated the therapeutic effect of Kuk B on high-fat diet/high-fructose (HFDFr)-induced insulin resistance and obesity in experimental rats.Materials and Methods: Rats were fed with either normal rat diet or HFDFr for 10 consecutive weeks. The groups that were fed with HFDFr received Kuk B (25 and 50 mg/kg) from the beginning of the 6th week to the 10th week. After treatment, the effect of Kuk B on body weight, food, water intake, insulin, blood glucose, serum biochemical parameters, hepatic oxidative stress (malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and proinflammatory cytokine (interleukin (IL)-6, interleukin (IL)-1β and tumor necrosis factor alpha (TNF-α)) levels was determined. Histopathological analysis of the liver tissues was also performed.Results: HFDFr-fed rats showed a significant increase in body weight, fasting blood glucose, insulin, lipid accumulation and liver function enzymes. In addition, HFDFr diet increased hepatic MDA, TNF-α, IL-1β and IL-6 and decreased hepatic SOD, CAT and GSH-Px activities. On the other hand, Kuk B significantly attenuated body weight, insulin resistance, lipid accumulation, oxidative stress and inflammation.Conclusion: These results indicated that Kuk B showed protective effect against HFDFr-induced metabolic disorders by downregulating lipid accumulation, oxidative stress and inflammatory factors.Keywords: kukoamine B, obesity, insulin resistance, high-fat diet, inflammation, oxidative stress

Objective To investigate the relationship between disease progression and microecology of tongue coating flora in patients with rheumatoid arthritis (RA). Methods A total of 30 tongue coating flora swabs were collected from 20 RA patients, including 10 in the active stage (OP group) and 10 in the remission stage (OR group), and 10 healthy individuals (OHC group).The total DNA extracted by rapid oral swab genomic DNA extraction kit was amplified with 16S rRNA and sequenced with high throughput Illumina MiSeq. The results were analyzed by microecology. Results There was no statistical difference in the diversity of 3 groups of samples (P>0.05). All the 3 groups of samples had their own unique genera, including 7 genera in OP group, 4 genera in OR group, and 10 genera in OHC group; The samples from 3 groups had the same composition of dominant flora, but obvious difference was seen in the composition of flora, mainly in p_Firmicutes, p_Actinobacteria, p_Episilonbacteraeota, p_Proteobacteria, g_Neisseria, g_Actinomycetes, g_Campylobacter, and g_Porphyromonas (all P < 0.05). Through PCA analysis and hierarchical cluster analysis, OP group and OHC group can gather together well with significant differences. Conclusion There are differences between the flora of tongue coating in RA patients and healthy individuals. The abundances of Actinomycetes and Campylobacter are increased, while those of Neisseria and Porphyromonas are decreased.

Peng Wan,1– 3,* Shiyun Cui,1– 3,* Zhenbao Ma,1– 3 Lin Chen,1– 3 Xiaoshen Li,1– 3 Ruonan Zhao,1– 3 Wenguang Xiong,1– 3 Zhenling Zeng1– 3 1Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou 510642, People’s Republic of China; 2National Laboratory of Safety Evaluation (Environmental Assessment) of Veterinary Drugs, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, People’s Republic of China; 3National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, South China Agricultural University, Guangzhou 510642, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wenguang Xiong Email xiongwg@scau.edu.cnZhenling Zeng Tel +86 20 85281204Fax +86 20 8528 4896Email zlzeng@scau.edu.cnPurpose: The plasmid-borne mobilized colistin resistance gene (mcr-1) was discovered in 2015. Subsequently, the rapid horizontal transfer of mcr-1 gene to diverse bacterial species poses a serious threat to public health, which urgently needs the introduction of novel antimicrobial strategies. Therefore, the purpose of this study is to sensitize bacteria to colistin and reduce the propagation of mcr-1 gene by curing mcr-1-harboring plasmid in Escherichia coli (E. coli) using the CRISPR-Cas9 system.Methods: Two sgRNAs specific to mcr-1 gene were designed and cloned into plasmid pCas9. The recombinant plasmid pCas9-mcr was transformed into E. coli carrying pUC19-mcr-1 or pHNSHP45, separately. The elimination efficiency in strains was evaluated by PCR and quantitative real-time PCR (qPCR). The antimicrobial susceptibility test was performed using the broth microdilution method.Results: In this study, we constructed the high copy number plasmid pUC19-mcr-1 and recombinant plasmid pCas9-m1 or pCas9-m2, which contain 20 nt or 30 nt sgRNA sequences targeted to mcr-1, respectively. PCR and qPCR results showed that mcr-1-harboring plasmids could be efficiently eliminated, and there was no significant correlation between sgRNA lengths and curing efficiency. However, when comparing restructured high copy number plasmid (pUC19-mcr-1) to natural resistance plasmid (pHNSHP45) in eliminating efficiency, we found that the content of plasmid backbone had an influence on efficiency. Furthermore, the conjugation assays verified that the engineered CRISPR-Cas9 system in bacteria or in bacteria genome can protect the recipient from plasmid-borne mcr-1 transfer via conjugation. Additionally, sequence analysis showed that three different types of defects in CRISPR-Cas9 system lead to escape mutants.Conclusion: We presented a method that only one plasmid-mediated CRISPR-Cas9 system can be used to efficiently resensitize E. coli to colistin. Moreover, this system provided a great potentiality to counteract the propagation of mcr-1 among bacterial pathogens.Keywords: sgRNA lengths, curing efficiency, backbone content, quantitative real-time PCR, conjugation assays, escape mutants

The yearly increase in government R&D investment and top-down national R&D investment allocation requires a more quantitative decision-making system that maximizes R&D performance and efficient budget allocation. Sound decision-making is necessary at both the selection stage and the pursuit stage in order to maximize limited national R&D resources. We study Korean smart farms as an example to examine national R&D investment from the various R&D actors (academia, industry, and research institutes) perspectives. The objective of our research is to evaluate the theoretical efficiency of R&D investment on specific technologies in smart farms and compare the results with expert opinions where the reality is reflected. To be specific, our study is to provide the quantitative approach in making decision among policymakers by reflecting the field experiences and opinions. We use a data envelopment analysis with an assurance region model, which integrates an analytic hierarchy process and a data envelopment analysis. The weights of output in DEA model by the R&D actors are similar to the overall weight by all actors, implying that investment allocation decisions in the smart farm sector are not significantly affected by the R&D actors. We realized that the relative efficiency of some R&D technologies increases after reflecting qualitative ideas of experts. In reality, it is necessary to invest in these technology groups, but they excluded from top-down decision-making. This also shows that a government’s top-down decision-making can distort its investment allocation. This study proposes a new approach to compensate for the difference between theoretical virtual prices and actual prices in data envelopment analysis. In particular, when comparing the only quantitative results on investment priorities with analysis results by additionally reflecting the opinions of experts in each sector, we found that the Korean government’s investment priorities in the smart farm field are considerably distorted. Therefore, this study is expected to be used as an alternative for policy makers to compensate for the quantitative distortion might be caused by top-down national R&D investment decisions.

Xiufeng Zhang,1–3 Dongdong Han,1–3 Pengfei Pei,1–3 Jie Hao,1–3 Yixing Lu,1–3 Peng Wan,1–3 Xianfeng Peng,4 Weibiao Lv,5 Wenguang Xiong,1–3 Zhenling Zeng1–3 1National Laboratory of Safety Evaluation (Environmental Assessment) of Veterinary Drugs, South China Agricultural University, Guangzhou 510642, People’s Republic of China; 2Guangdong Provincial Key Laboratory of Veterinary Drugs Development and Safety Evaluation, South China Agricultural University, Guangzhou 510642, People’s Republic of China; 3National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, South China Agricultural University, Guangzhou 510642, People’s Republic of China; 4Guangzhou Insighter Biotechnology Co., Ltd, Guangzhou 510642, People’s Republic of China; 5Department of Clinical Laboratory, Shunde Hospital, Southern Medical University (The First People’s Hospital of Shunde), Foshan 528000, People’s Republic of ChinaCorrespondence: Wenguang Xiong; Zhenling ZengSouth China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, People’s Republic of ChinaTel +86 20-85281204Fax +86 20-85284896Email xiongwg@scau.edu.cn; zlzeng@scau.edu.cnBackground: Bacterial infections cause a serious public health crisis due to the emergence of resistance towards multiple conventional antibacterial drugs. In particular, multidrug-resistant (MDR) Enterococcus faecium which belongs to “ESKAPE” organisms is causing significant problems worldwide. Hence, there is an urgent need to find alternative therapies. Recently, substituted benzene guanidine compounds have been used as lead structures to discover new promising drugs in both synthetic and medicinal chemistry.Purpose: Here we investigated the antimicrobial activity of a new substituted benzene guanidine analog, isopropoxy benzene guanidine, against Enterococci.Material and methods: The isopropoxy benzene guanidine was synthesized by Guangzhou Insighter Biotechnology Co., Ltd and tested on both reference bacterial strain and 32 clinical MDR Enterococci strains. The in vitro antibacterial activity was evaluated by microdilution method and kill kinetic assays. The potential antibacterial mechanism was measured by fluorescence spectrometry using fluorescent membrane potential probe 3, 3-diethyloxacarbocyanine iodide (DiOC2 (3)).Results: Isopropoxy benzene guanidine exhibited potent bactericidal activity against both reference strain and MDR Enterococci isolates. The minimum inhibitory concentration (MIC) range for isopropoxy benzene guanidine was 1–4 μg/mL. Minimum bactericidal concentration (MBC) was about 2-8-fold of its MIC values. Time-kill studies showed that isopropoxy benzene guanidine provided superior bactericidal effect against reference and MDR strains within 12 hrs at 2×MIC. Furthermore, isopropoxy benzene guanidine could cause a large reduction in the magnitude of the generated membrane potential compared to that of the untreated cells.Conclusion: The present study highlights the potent bactericidal activity of isopropoxy benzene guanidine on Enterococci by disrupting the cell membrane potential. These findings demonstrate that isopropoxy benzene guanidine may be a good chemical lead for further medicinal chemistry and pharmaceutical development and could be used as a therapeutic agent for infectious diseases caused by MDR Enterococci.Keywords: isopropoxy benzene guanidine, MDR Enterococci, DiOC2, 3, cell membrane potential, bactericidal activity