The microstructure and damage evolution behavior of IN718 alloy prepared by selective laser melting (SLM, referred to as SLM IN718 alloy) under different heat treatment processes were studied using optical microscopy, scanning electron microscopy, electron backscatter diffraction, microhardness tests, and other material tests, combined with digital image correlation. The heat treatment of the SLM IN718 alloy achieved different degrees of recrystallization and transformed the microstructure from dendritic crystals to bulk crystals with the precipitation of a large number of γ′ and γ′′ phases. As the heat-treatment temperature was increased, the δ phase of the SLM IN718 alloy was precipitated and then dissolved, the microhardness and strength of SLM IN718 alloy increased and then decreased, the elongation decreased and then increased, and toughness fracture was the main fracture mode. Therefore, heat treatment can be used to alter the relative proportions of recrystallized and substructured grains in the SLM IN718 alloy, thereby modulating its overall mechanical properties. After heat treatment, the damage factor entered the rapid damage stage earlier, and the critical damage factor increased as the critical plastic strain was increased. Further, the damage evolution equations of the SLM IN718 alloy under different heat treatment processes were established. IN IN71 SLM, (SLM microscopy diffraction tests correlation γ phases heattreatment dissolved mode Therefore properties earlier Further established IN7

Abstract Background Varicocele is an abnormal expansion of the pampininias venous plexus in the scrotum, resulting in impaired sperm production and reduced sperm quality. The exact pathophysiological mechanism leading to varicocele-related infertility has not been fully elucidated. Although treatable, varicocele may lead to male infertility. Objective To investigate the relationship between semen parameters, serum InhB and INSL-3 levels, and the degree of varicocele in male patients. Methods Serum InhB and INSL-3 were detected. To evaluate the relationship between semen parameters and serum InhB and INSL-3 levels. To evaluate the value of semen parameters and serum InhB and INSL-3 levels in distinguishing disease severity in patients with varicocele. Results Serum INSL-3 in patients with varicocele decreased with the severity of the disease. Serum INSL-3 was positively correlated with total sperm count and frequency of normal sperm morphology. There was a weak correlation between serum InhB and semen volume, concentration, and total sperm. Patients with different disease severity were similar within the groups, with partial overlap or similarity between varicocele Grade I and Grade II, and significant differences between Grade III and Grade I and II. Semen volume, concentration, total sperm, normal sperm morphology, and serum InhB and INSL-3 levels could distinguish the degree of varicocele. Conclusion Semen parameters and the combination of serum InhB and INSL-3 levels in patients with varicocele are closely related to the severity of the disease. Serum INSL-3 is expected to be a potential biomarker for early clinical intervention. scrotum quality varicocelerelated elucidated treatable INSL3 INSL 3 INSL- detected morphology volume concentration groups II intervention

Abstract Background: Acid-sensing ion channel 1a (ASIC1a) plays a critical role in physiological and pathological processes. To further elucidate the biological functions of ASICs and their relationships with disease occurrence and development, it is advantageous to investigate and develop additional regulatory factors for ASICs. Methods: In this study, cation exchange chromatography was used to separate seven chromatographic components from Naja naja atra venom. Capillary electrophoresis was employed to detect that Ⅶ peak component containing a main protein Ⅶ-2, which could bind to ASIC1a. The analgesic effects of Ⅶ-2 protein were determined using hot plate methods, and ASIC1a expression in spinal cord tissue from rats with inflammatory pain was detected using western blot. Results: The purified Ⅶ-2 protein named Naja naja atra venom-Ⅶ-2 (NNAV-Ⅶ-2) was obtained by Sephadex G-50 gel filtration, which exhibited a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular weight of 6.7 kD. Remarkably, the NNAV-Ⅶ-2 protein demonstrated a significant analgesic effect and downregulated ASIC1a expression in the spinal cord tissue of rats with inflammatory pain. Conclusions: The analgesic mechanism of the NNAV-Ⅶ-2 protein may be associated with its binding to ASIC1a, consequently downregulating ASIC1a expression in neural tissues. Background Acidsensing Acid sensing ASICa ASIC (ASIC1a processes development Methods study venom Ⅶ2, Ⅶ2 2, 2 Ⅶ- methods blot Results venomⅦ2 venom-Ⅶ- NNAVⅦ2 NNAV (NNAV-Ⅶ-2 G50 G 50 G-5 filtration sulfatepolyacrylamide sulfate polyacrylamide 67 6 7 6. kD Remarkably NNAV-Ⅶ- Conclusions tissues venomⅦ venom-Ⅶ NNAVⅦ (NNAV-Ⅶ- G5 5 G- NNAV-Ⅶ venom- (NNAV-Ⅶ NNAV- (NNAV- (NNAV

ABSTRACT Objectives: We examined the expression of Lnc-ZFAS1 in osteosarcoma and comprehensively evaluated its effects on osteosarcoma in vitro and vivo. Moreover, we revealed the regulatory mechanism between Lnc-ZFAS1 and miR-520b/miR-520e-mediated RHOC and provided a novel clue for ameliorating osteosarcoma. Method: The expression of Long non-coding RNA Zinc Finger Antisense 1 (LncRNA ZFAS1) osteosarcoma tissues and normal tissues in the TCGA database was analyzed. Then, LncRNA ZFAS1 expression was further verified in clinical samples and osteosarcoma cell lines (U2OS and KHOS), as well as the human osteoblast cell line hFOB1.19 by qRT-PCR. Thereafter, LncRNA ZFAS1 was overexpressed or silenced to explore its effects on cell proliferation, apoptosis, migration, invasion, and Epithelial-Mesenchymal Transition (EMT). The fundamental mechanism through which Lnc-ZFAS1 affects osteosarcoma progression was further investigated and verified. Results: We found that LncRNA ZFAS1 was upregulated in osteosarcoma, and Lnc-ZFAS1 overexpression facilitated osteosarcoma cells proliferation, migration, invasion and EMT, while Lnc-ZFAS1 silence exerted reverse influence. Mechanistically, Lnc-ZFAS1 functionally acted as a sponger of microRNA-520b (miR-520b) and micro-RNA-520e (miR-520e) to up-regulate Ras Homologue C (RHOC). In addition, depleted Lnc-ZFAS1 restrained osteosarcoma cells proliferation, migration, and invasion, which could be rescued by RHOC overexpression. Lnc-ZFAS1 was upregulated in osteosarcoma and Lnc-ZFAS1 could exert promoted impact upon osteosarcoma cells proliferation, migration, invasion, and EMT in vitro. Conclusions: Lnc-ZFAS1 acted sponger of miR-520b and miR-520e to promote RHOC, indicating that Lnc-ZFAS1/miR-520b/RHOC and Lnc-ZFAS1/miR-520e/RHOC axes might serve as potential therapeutic strategies against osteosarcoma. Objectives LncZFAS1 LncZFAS Lnc ZFAS Lnc-ZFAS vivo Moreover miR520b/miR520emediated miR520bmiR520emediated miRbmiRemediated miR 520b/miR 520e mediated 520b b e Method noncoding non coding analyzed Then U2OS UOS U OS KHOS, KHOS , KHOS) hFOB119 hFOB hFOB1 19 hFOB1.1 qRTPCR. qRTPCR qRT PCR. PCR qRT-PCR Thereafter proliferation apoptosis migration EpithelialMesenchymal Epithelial Mesenchymal EMT. . (EMT) Results influence Mechanistically microRNA520b microRNAb microRNA miR520b miRb (miR-520b microRNA520e microRNAe micro miR520e miRe (miR-520e upregulate up regulate RHOC. (RHOC) addition Conclusions LncZFAS1/miR520b/RHOC LncZFAS1miR520bRHOC LncZFASmiRbRHOC ZFAS1/miR 520b/RHOC LncZFAS1/miR520e/RHOC LncZFAS1miR520eRHOC LncZFASmiReRHOC 520e/RHOC miR520emediated emediated bmiR 520bmiR hFOB11 hFOB1. (EMT (RHOC bRHOC ZFAS1miR ZFASmiR 520bRHOC eRHOC 520eRHOC miRemediated

ABSTRACT Purpose: To investigate the role of cyanidin-3-O-glucoside (C3G) in renal ischemia/reperfusion (I/R) injury and the potential mechanisms. Methods: Mouse models were established by clamping the left renal vessels, and in vitro cellular models were established by hypoxic reoxygenation. Results: Renal dysfunction and tissue structural damage were significantly higher in the I/R group. After treatment with different concentrations of C3G, the levels of renal dysfunction and tissue structural damage decreased at different levels. And its protective effect was most pronounced at 200 mg/kg. The use of C3G reduced apoptosis as well as the expression of endoplasmic reticulum stress (ERS)-related proteins. Hypoxia/reoxygenation (H/R)-induced apoptosis and ERS are dependent on oxidative stress in vitro. In addition, both AG490 and C3G inhibited the activation of JAK/STAT pathway and attenuated oxidative stress, ischemia-induced apoptosis and ERS. Conclusions: The results demonstrated that C3G blocked renal apoptosis and ERS protein expression by preventing reactive oxygen species (ROS) production after I/R via the JAK/STAT pathway, suggesting that C3G may be a potential therapeutic agent for renal I/R injury. Purpose cyanidin3Oglucoside cyanidinOglucoside cyanidin 3 O glucoside CG C G (C3G ischemiareperfusion ischemia reperfusion IR I R (I/R mechanisms Methods vessels reoxygenation Results group 20 mgkg mg kg mg/kg ERSrelated related proteins Hypoxiareoxygenation Hypoxia H/Rinduced HRinduced H/R induced H addition AG AG49 JAKSTAT JAK STAT ischemiainduced Conclusions ROS (ROS Oglucoside 2 Rinduced HR AG4

Abstract African swine fever (ASF) is an important viral disease of swine caused by the African swine fever virus (ASFV), which threatens swine production profoundly. To better understand the gene expression changes when pig infected with ASFV, RNA sequencing was performed to characterize differentially expressed genes (DEGs) of six tissues from Kenya domestic pigs and Landrace × Yorkshire (L/Y) pigs infected with ASFV Kenya1033 in vivo. As results, a total of 209, 522, 34, 505, 634 and 138 DEGs (q-value < 0.05 and |Log2foldchange| values >2) were detected in the kidney, liver, mesenteric lymph node, peripheral blood mononuclear cell, submandibular lymph node and spleen, respectively. The expression profiles of DEGs shared in the multiple tissues illustrated variation in regulation function in the different tissues. Functional annotation analysis and interaction of proteins encoded by DEGs revealed that genes including IFIT1, IFITM1, MX1, OASL, ISG15, SAMHD1, IFINA1, S100A12 and S100A8 enriched in the immune and antivirus pathways were significantly changed when the hosts were infected with ASFV. The genes mentioned could play crucial roles in the process of the reaction to non-lethal ASF infection, which may will help to improve the ASF tolerance in the pig population through molecular breeding strategies. (ASF , (ASFV) profoundly (DEGs L/Y LY L Y (L/Y Kenya103 vivo results 209 522 34 505 63 13 qvalue q value 005 0 05 0.0 Log2foldchange Logfoldchange Log foldchange |Log2foldchange >2 2 kidney liver cell spleen respectively IFIT1 IFIT IFITM1 IFITM MX1 MX OASL ISG15 ISG SAMHD1 SAMHD IFINA1 IFINA SA S A S100A1 S100A nonlethal non lethal infection strategies (ASFV Kenya10 20 52 3 50 6 1 00 0. > ISG1 Kenya1 5

RESUMEN La contaminación por neblina, caracterizada principalmente por la baja visibilidad, es uno de los principales problemas ambientales a los que se enfrenta China en la actualidad. El pronóstico exacto de la neblina es útil para la aplicación de medidas preventivas orientadas a controlar emisiones de contaminantes atmosféricos y aliviar la contaminación por neblina. Sin embargo, no es fácil predecir con precisión los acontecimientos de baja visibilidad causados por neblina, lo cual requiere no sólo un pronóstico riguroso de los factores meteorológicos, sino también una actualización detallada y en tiempo real de los inventarios de emisiones de las fuentes. Con el fin de obtener una herramienta de predicción fiable, se estudian varios métodos populares de aprendizaje automático, como la máquina vectorial de apoyo (SVM, por su sigla en inglés), el k-vecino más próximo, el bosque aleatorio y varios métodos de aprendizaje profundo. A partir de los principales factores que influyen en la visibilidad, se analizan las relaciones entre velocidad y dirección del viento, temperatura, humedad y visibilidad. El método de aprendizaje automático se utiliza para entrenamiento y pronóstico. La precisión de estos métodos para pronosticar la visibilidad se verifica mediante varios parámetros (error de raíz cuadrada media, error absoluto medio y error porcentual absoluto medio). Los resultados muestran que: 1) entre todos los parámetros meteorológicos, la velocidad del viento puede reflejar mejor la variación de la visibilidad; 2) el rendimiento de las redes neuronales recurrentes de memoria a largo-corto plazo (LSTM RNN) y el método de unidades recurrentes cerradas (GRU) en la predicción de visibilidad a corto plazo (es decir, 1, 3 y 6 h) es casi igual; 3) el método clásico de aprendizaje automático (SVM) funciona bien para el pronóstico de visibilidad a medio y largo plazo, y 4) el método de aprendizaje automático también tiene cierto grado de precisión en el pronóstico, incluso en un periodo de tiempo más largo (p. ej., 72 h).

ABSTRACT Haze pollution, mainly characterized by low visibility, is one of the main environmental problems currently faced by China. Accurate haze forecasts facilitate the implementation of preventive measures to control the emission of air pollutants and thereby mitigate haze pollution. However, it is not easy to accurately predict low visibility events induced by haze, which requires not only accurate prediction for weather elements, but also refined and real-time updated source emission inventory. In order to obtain reliable forecasting tools, this paper studies the usability of several popular machine learning methods, such as support vector machine (SVM), k-nearest neighbor, and random forest, as well as several deep learning methods, on visibility forecasting. Starting from the main factors related to visibility, the relationships between wind speed, wind direction, temperature, humidity, and visibility are discussed. Training and forecasting were performed using the machine learning methods. The accuracy of these methods in visibility forecasting was confirmed through several parameters (i.e., root-mean-square error, mean absolute error, and mean absolute percentage error). The results show that: (1) among all meteorological parameters, wind speed was the best at reflecting the visibility change patterns; (2) long short-term memory recurrent neural networks (LSTM RNN), and gated recurrent unit (GRU) methods perform almost equally well on short-term visibility forecasts (i.e., 1, 3, and 6 h); (3) a classical machine learning method (i.e., the SVM) performs well in mid- and long-term visibility forecasts; (4) machine learning methods also have a certain degree of forecast accuracy even for long time periods (e.g., 7 2h).

RESUMO Introdução O tênis é um esporte de rede que exige força, resistência, agilidade, velocidade, habilidade e outras demandas físicas dos participantes. Os atletas de tênis estão sujeitos a lesões em seus treinamentos diários, afetando o seu desempenho competitivo. Objetivo Explorar a correlação entre o local das lesões e a causa das lesões nos jogadores de tênis. Métodos Os jogadores de tênis foram selecionados como os objetos de pesquisa. Neste estudo, foi utilizado um questionário para registrar as lesões no tênis. Através de método estatístico, resumiu-se e analisou-se o tratamento de reabilitação dos jogadores de tênis após as lesões. Resultados As principais formas de lesões esportivas graves causadas pelos tenistas foram lesões agudas e moderadas. As fraturas mais comuns que ocorrem são principalmente as ligamentares e tendinosas: lesão ou laceração do ligamento articular, lesão ligamentar muscular, bursite e contusão ligamentar de tecidos moles. As articulações mais envolvidas são joelhos, tornozelos, cotovelos, pulsos e ombros. Conclusão As causas comuns das lesões esportivas nos tenistas são uma união de imprecisão dos movimentos técnicos, atividades de força muscular insuficientes e força insuficiente para atividades técnicas preparatórias. Na prática, recomenda-se aos atletas corrigir e padronizar seus movimentos. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

ABSTRACT Introduction Tennis is a network sport that requires strength, endurance, agility, speed, skill, and other physical demands from participants. Tennis athletes are subject to injuries in their daily training, affecting their competitive performance. Objective Explore the correlation between the location of injuries and the cause of injuries in tennis players. Methods Tennis players were selected as the research subjects. In this study, a questionnaire was used to record tennis injuries. The rehabilitation treatment of tennis players after injuries was summarized and analyzed through statistical methods. Results The main forms of serious sports injuries caused by tennis players were acute and moderate injuries. The most common fractures are mainly ligament and tendon: joint ligament injury or tear, muscle ligament injury, bursitis, and soft tissue ligament contusion. The joints most commonly involved are knees, ankles, elbows, wrists, and shoulders. Conclusion The common causes of sports injuries in tennis players are the inaccuracy of technical movements, insufficient muscular strength, and insufficient strength for preparatory technical activities. In practice, it is recommended that athletes correct and standardize their movements. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMEN Introducción El tenis es un deporte de red que exige fuerza, resistencia, agilidad, velocidad, habilidad y otras exigencias físicas a los participantes. Los atletas de tenis están sujetos a lesiones en su entrenamiento diario, lo que afecta a su rendimiento competitivo. Objetivo Explorar la correlación entre la localización de las lesiones y la causa de las mismas en los tenistas. Métodos Se seleccionaron jugadores de tenis como sujetos de la investigación. En este estudio, se utilizó un cuestionario para registrar las lesiones de tenis. Mediante un método estadístico, se resumió y analizó el tratamiento de rehabilitación de los tenistas tras las lesiones. Resultados Las principales formas de lesiones deportivas graves causadas por los tenistas fueron las lesiones agudas y las moderadas. Las fracturas más comunes que se producen son principalmente las de ligamentos y tendones: lesión o desgarro de ligamentos articulares, lesión de ligamentos musculares, bursitis y contusión de ligamentos de tejidos blandos. Las articulaciones más comúnmente afectadas son las rodillas, los tobillos, los codos, las muñecas y los hombros. Conclusión Las causas comunes de las lesiones deportivas en los tenistas son la unión de la inexactitud de los movimientos técnicos, las actividades de fuerza muscular insuficientes y la fuerza insuficiente para las actividades técnicas preparatorias. En la práctica, se recomienda que los deportistas corrijan y normalicen sus movimientos. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

ABSTRACT Purpose: This study aimed to develop a minimally invasive surgical procedure for laminar lift and posterior cervical laminoplasty via the intermuscular approach using a canine model. Methods: Six Alaskan dogs were used for developing the surgical approach. The bilateral laminae of C3-7 were cut with an ultrasonic osteotome and fixed with bilateral plates to maintain the lamina lifting and reshape a wider spinal canal. The important structures, such as ligaments, supraspinous ligaments, interspinous ligaments, and ligamentum flavum were preserved. The therapeutic effect was evaluated by preoperative and postoperative imaging results and neck mobility. Results: The surgical procedures were all successfully performed in the 6 animals. All the dogs survived well within 1 year of postoperative follow-up. The postoperative neck mobility was as good as the preoperative one. Computed tomography results showed that the anteroposterior diameter of the spinal canal was successfully enlarged and maintained well. Conclusions: The minimally invasive surgical procedure for laminar lift and posterior cervical laminoplasty via the intermuscular approach was feasible in a canine model, which might be applied in clinical practice.

Inconel 718 alloy which was prepared using selective laser melting (SLM) forming technology underwent various annealing treatments. Optical microscope, scanning electron microscope, electron backscattered diffraction and MTS testing machine were used to study the microstructure, mechanical and fatigue properties of Inconel 718 alloy melted by selective laser melting under different annealing treatments. The results show that the microstructure of Inconel 718 alloy after homogenization and doubleaging annealing is the most obvious, and the microstructure of the alloy is mainly recrystallized. The microstructure contains a large number of annealing twins and the grain boundaries are flat. The yield strength, tensile strength and microhardness of Inconel 718 alloy formed by selective laser melting after different annealing treatments are greatly increased, while the elongation after fracture is significantly decreased. The fatigue properties of Inconel 718 alloy after double aging annealing and solution dual aging annealing are improved slightly, while the fatigue properties after uniform double aging annealing are decreased slightly.

Eccrine sweat glands (ESGs) perform critical functions in temperature regulation in humans. Foxa1 plays an important role in ESG maturation and sweat secretion. Its molecular mechanism, however, remains unknown. This study investigated the expression of Foxa1 and Na-K-ATPase (NKA) in rat footpads at different development stages using immunofluorescence staining, qRT-PCR, and immunoblotting. Also, bioinformatics analysis and Foxa1 overexpression and silencing were employed to evaluate Foxa1 regulation of NKA. The results demonstrated that Foxa1 was consistently expressed during the late stages of ESGs and had a significant role in secretory coil maturation during sweat secretion. Furthermore, the mRNA abundance and protein expression of NKA had similar accumulation trends to those of Foxa1, confirming their underlying connections. Bioinformatics analysis revealed that Foxa1 may interact with these two proteins via binding to conserved motifs in their promoter regions. Foxa1 gain-of-function and loss-of-function experiments in Foxa1-modified cells demonstrated that the activities of NKA were dependent on the presence of Foxa1. Collectively, these data provided evidence that Foxa1 may influence ESG development through transcriptional regulation of NKA expression.

ABSTRACT Oil is a non-renewable resource, and it is essential to improve oil recovery. Surfactant flooding is a key technology in enhancing oil recovery. The present work aims to develop a new surfactant to the combination flooding system in oil displacement. The Aminosulfonic Acid Gemini Surfactant was prepared by using Hexadecylamine, 1-Butanaminium,N,N,N-tributyl-,bromide (1:1) and Sodium 2-chloroethyl sulfonate. The AminoSulfonic Acid Gemini Surfactant Sodium N,N’-Dihexadecyl-2-propanol- 1,3-ethylsulfonate SNNDPE) has better reaction temperature at 80℃. The best reaction time of SNNDPE is 12 h. Next, We made a characterization of SNNDPE. A new gemini surfactant was developed, which is beneficial to the research of material science. Then the performance of the new Gemini surfactant was tested. The surface tension tests suggested that the newly synthesized Aminosulfonic Acid Gemini Surfactant has higher surface activity. The resistance temperature of the Aminosulfonic Acid Gemini Surfactant solution is 80℃. Moreover, the oil-water interfacial tension experiment suggested that the minimum aqueous solution of the combination flooding system reaches an ultra-low level of 9.4 × 10−4 mN/m. The combination flooding system experiment results can be considered as a guidance in the design and studies of the combination flooding system for a selective.

ABSTRACT To accurately measure sowing depth, we designed a multisensor data fusion algorithm based on the Kalman filter and applied it to a dual-channel ultrasonic sowing depth measurement system. We installed two ultrasonic sensors on the frame above the depth limit wheel and the soil covering-compacting wheel. The terrain-like plate was installed on the central axis of the depth limit wheel to simulate the surface relief and calculate the seeding depth value. The Kalman filter algorithm filters out the clutter generated when the two ultrasonic sensors collect the depth data and then fuses the two ultrasounds of the filtered data. The test shows that when the vehicle speed is 4 km/h , the fluctuation ranges of the collected data of ultrasonic 1 and 2 are 45-60 mm and 35-70 mm , respectively, and the fluctuation ranges of the filtered data are 52-58 mm and 48-60 mm , respectively. After data fusion processing, the data fluctuate in a small range of approximately 55 mm . When the vehicle speed is 6 km/h and 8 km/h, the obtained results have the same trend as the experimental results at 4 km/h . The results show that the sowing depth measurement system is effective.

Abstract Evaluation of volatile aroma compounds from Chinese wild grape species aimed to explore new grape species via fruit aroma. In this study, the volatile organic compounds (VOCs) from ten grape cultivars belonging to nine different grape species were detected, by using headspace solid-phase microextraction with GC-MS. Chinese wild grape species were studied along with table grape (Muscat Hamburg, Concord) and wine grape (Cabernet Sauvignon) cultivars belonging to different origins. Total forty-five volatile compounds were detected, upon which several VOCs including pentanoic acid, cyclobutyl ester, maltol, (+)-neoisomenthol, and 8-methyloctahydrocoumarin were detected in Chinese wild grapes that have not been detected in other Vitis species. The results revealed that C6 volatile compounds were pre-dominant VOCs. Esters were found abundant in V. labrusca (Concord) and terpenoids were dominant in V. vinafera (Hamburg Muscat). The principal component analysis showed that Chinese wild grape cultivars V. yeshanensis, V. facifolia, V. davidii, V. betulifolia V. heyneana, V. amurensis and V. adenoclada are correlated with wine grape cultivar ‘Cabernet Sauvignon’, while table grape cultivars are distributed separately. These results can provide a reference for the utilization of new germplasm for specific purposes.

Abstract Transfection efficiency was estimated to optimize the conditions for RNA interference (RNAi), including transfection time, validity, and nucleic acid concentration and type, using the EZ Trans Cell Reagent, a cationic polymer. An shRNA against GFP was designed and transfected into cells using the EZ transfection reagent. The shRNA significantly decreased the expression of GFP. In addition, pre-diluted transfection reagent at room temperature and small nucleic acids increased the transfection efficiency, which peaked at 24 h. Compared with circular nucleic acids, linear nucleic acids showed higher transfection efficiency and a higher genome integration rate. We optimized cationic polymer-mediated RNAi conditions, and our data will be useful for future RNAi studies.