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Objectives: This study aimed to assess and compare the effect of an 8-week high-intensity interval training (HIIT) or moderate-intensity continuous training (MICT) programme on body composition and cardiovascular metabolic outcomes of sedentary adolescents in China. Methods: Eighteen sedentary normal-weight adolescents (age: 18.5 ± 0.3 years, 11 females) were randomized into three groups. HIIT group protocol consisted of three sessions/week for 8-week of "all out" sprints to reach 85%-95% of HRmax, and MICT group protocol undertook three sessions/week for 8-week of continuous running to reach 65%-75% of HRmax. The control group resumed normal daily activities without any intervention. Blood pressure and body composition were measured, and fasting blood samples were obtained at baseline and 48 h post-trial. Mixed-design ANOVA analysis was employed followed by post hoc t-tests and Bonferroni alpha-correction was used to evaluate interaction, between-group, and within-group differences, respectively. Results: Results indicated that HIIT and MICT similarly affected body fat mass (p = 0.021, ES = 0.19; p = 0.016, ES = 0.30, respectively), body fat percentage (p = 0.037, ES = 0.17; p = 0.041, ES = 0.28, respectively), visceral fat area (p = 0.001, ES = 0.35; p = 0.003, ES = 0.49, respectively) of body composition. A positive outcome was observed for waist/hip ratio (p = 0.033, ES = 0.43) in HIIT, but not MICT (p = 0.163, ES = 0.33). No significant differences were found between groups for any clinical biomarkers. However, pairwise comparison within the group showed a significant decrease in systolic blood pressure (p = 0.018, ES = 0.84), diastolic blood pressure (p = 0.008, ES = 1.76), and triglyceride (p = 0.004, ES = 1.33) in HIIT, but no significant differences were found in the MICT and Control group. Conclusion: Both 8-week HIIT and MICT programmes have similar positive effects on reducing body fat mass, fat percentage, and visceral fat area. However, sedentary adolescents may have limited scope to decrease insulin resistance after these 8-week interventions. Notably, the 8-week HIIT intervention was highly effective in increasing cardiometabolic health compared to the MICT. The exercise intensity threshold value and metabolic outcomes of high-intensity interval sprints should be explored further to extend the long-term benefit in this cohort.
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Individuals' continuous success in competitive interactions with conspecifics strongly affects their social hierarchy. Medial prefrontal cortex (mPFC) is the key brain region mediating both social competition and hierarchy. However, the molecular regulatory mechanisms underlying the neural ensemble in the mPFC remains unclear. Here, we demonstrate that in excitatory neurons of prelimbic cortex (PL), lncRNA Sera remodels the utilization of Pkm Exon9 and Exon10, resulting in a decrease in the Pkm1/2 ratio in highly competitive mice. By employing a tet-on/off system, we disrupt or rebuild the normal Pkm1/2 ratio by controlling the expression of Pkm2 in PL excitatory neurons. We find that long-term Pkm2 modulation induces timely competition alteration and hysteretic rank change, through phosphorylating the Ser845 site of GluA1. Together, this study uncovers a crucial role of lncRNA Sera/Pkm2 pathway in the transition of social competition to rank by remodeling neural ensemble in mPFC.
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BACKGROUND: A precise model for predicting outcomes is needed to guide perioperative management. With the developments of liver transplantation (LT) discipline, previous models may become inappropriate or noncomprehensive. Thus, we aimed to develop a novel model integrating variables from donors and recipients for quick assessment of transplant outcomes. METHODS: The risk model was based on Cox regression in a randomly selected derivation cohort and verified in a validation cohort. Perioperative data and overall survival were compared between stratifications grouped by X-tile. Receiver operating characteristic curve and decision curve analysis were used to compare the models. Violin and raincloud plots were generated to present post-LT complications distributed in different stratifications. RESULTS: Overall, 528 patients receiving LT from 2 centers were included with 2/3 in the derivation cohort and 1/3 in the validation cohort. Cox regression analysis showed that cold ischemia time (CIT) (P=0.012) and the Model for End-Stage Liver Disease (MELD) (P=0.007) score were predictors of survival. After comparison with the logarithmic models, the primitive algorithms of CIT and MELD were defined as the CIT-MELD Index (CMI). CMI was stratified by X-tile (grade 1 ≤1.06, 1.06< grade 2 ≤1.87, grade 3 >1.87). In both cohorts, CMI performed better in calculating transplant outcomes than the balance of risk score, including perioperative incidents and prevalence of complications. CONCLUSIONS: Model integrating variables from graft and recipient made the prediction more accurate and available. CMI provided new sight in outcome evaluation and risk factor management of LT.
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The digestion rate of steamed soybean (DRSS), protease activity of koji (PAK) and formaldehyde nitrogen content of moromi (FNCM) are key indicators to monitor soy sauce production. Currently, monitoring these indicators relies on workers' experience, which can sometimes lead to low material utilization rates and even fermentation failures. Near-infrared spectra were collected during soybean steaming, as well as koji and moromi fermentation, using miniature fiber spectrometers. These spectra were optimized using four pretreatment methods, and regression models were constructed using PLS, iPLS, and Si-PLS. The evaluation of models in prediction sets was based on the correlation coefficient (Rp) and root mean square error (RMSEP). Results indicated that Rp = 0.9327, RMSEP = 4.37% for DRSS, Rp = 0.9364, RMSEP = 228 U/g for PAK, and Rp = 0.9237, RMSEP =0.148 g/100 mL for FNCM were obtained. The above results coupling with validation experiments demonstrated that the developed in-situ and real-time spectroscopy system could ensure high-quality soy sauce production.
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Existing studies have examined unhealthy food packaging information, mainly focusing on aspects such as the content, color, and text, whilst paying less attention to the boundaries of information. This paper investigates unhealthy foods through three experiments, revealing that the presence (vs. absence) of packaging information boundaries on unhealthy foods has a negative impact on consumers' purchasing intentions (p = 0.040) (Experiment 1). The feeling of constraint mediates this effect (ß = -0.078, CI: [-0.1911, -0.0111]) (Experiment 2). Additionally, consumers with an independent self-construal exhibit reduced purchasing intentions when unhealthy food packaging information boundaries are present (vs. absent) (p < 0.001), whereas those with an interdependent self-construal show increased purchasing intentions under the same conditions (p = 0.024) (Experiment 3). This paper reveals the psychological mechanism and boundary conditions of unhealthy food packaging information boundaries affecting consumers' purchasing intention and provides practical inspiration for government policy-making related to unhealthy food packaging.
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BACKGROUND: Studies investigating the relationship between flavonoid intake and chronic kidney disease (CKD) are limited. This study investigated the association between daily flavonoid intake and CKD in US adults by using data for 2007-2008, 2009-2010, and 2017-2018 from the National Health and Nutrition Examination Survey (NHANES) database. METHODS: This study employed a cross-sectional design and used data from three cycles of the continuous NHANES: 2007-2008, 2009-2010, and 2017-2018. NHANES researchers collected data related to consumption of various food and beverages from participants by employing 24-h dietary recall questionnaires. CKD is defined as an estimated glomerular filtration rate of < 60 mL/min/1.73m2 or a urine albumin-to-creatinine ratio of ≥ 30 mg/g. RESULTS: The odds ratios (OR) for CKD risk in the second (Q2), third (Q3), and fourth (Q4) quartiles of total flavonoid intake, compared with that in the first (Q1) quartile, were 0.780 (95% CI: 0.600, 1.015), 0.741 (95% CI: 0.573, 0.957), and 0.716 (95% CI: 0.554, 0.925), respectively (with a P value for the trend of 0.040). According to the restricted cubic spline analysis, total flavonoid intake exhibited a non-linear relationship with CKD risk (P < 0.001). CONCLUSION: Our findings suggest that a potential J-shaped relationship was observed between total flavonoid consumption and CKD risk, with an inflection point at 69.58 mg/d. Our study indicates that a moderate intake of flavonoids may confer renal benefits which may offer novel strategies for CKD treatment.
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Flavonoides , Encuestas Nutricionales , Insuficiencia Renal Crónica , Humanos , Flavonoides/administración & dosificación , Insuficiencia Renal Crónica/epidemiología , Masculino , Femenino , Persona de Mediana Edad , Adulto , Estudios Transversales , Estados Unidos/epidemiología , Dieta , Anciano , Tasa de Filtración Glomerular , Factores de Riesgo , Adulto JovenRESUMEN
Nitrate pollution poses significant threats to both aquatic ecosystems and human well-being, particularly due to eutrophication and increased risks of methemoglobinemia. Conventional treatment for nitrate-contaminated wastewater face challenges stemming from limited availability of carbon sources and the adverse impacts of toxins on denitrification processes. This study introduces an innovative Intimately Coupled Photocatalysis and Biodegradation (ICPB) system, which utilizes Ag3PO4/Bi4Ti3O12, denitrifying sludge, and polyurethane sponge within an anoxic environment. This system demonstrates remarkable efficacy in simultaneously removing bio-recalcitrant organic compounds (such as sulfamethoxazole) and nitrates, surpassing standalone treatment methods. Optimally, the ICPB achieves complete removal of sulfamethoxazole, along with 87.7 % removal of DOC, and 81.8 % reduction in nitrate levels. Its ability to sustain pollutant removal and biological activity over multiple cycles can be attributed to the special formation of biofilm and mineralization of sulfamethoxazole, minimizing both photocatalytic damage and toxic inhibitory effects on microbes. The dominant microbial genera of ICPB system included Castellaniella, Acidovorax, Raoultella, Giesbergeria, and Alicycliphilus. Additionally, the study sheds light on a potential mechanism for the concurrent treatment of recalcitrant organics and nitrates by the ICPB system, presenting a novel and highly effective approach for addressing biologically resistant wastewater.
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Springtails (Collembola) stand as one of the most abundant, widespread, and ancient terrestrial arthropods on earth. However, their evolutionary history and deep phylogenetic relationships remain elusive. In this study, we employed phylogenomic approaches to elucidate the basal relationships among Collembola. We sampled whole-genome data representing all major collembolan lineages in proportion to their known diversity. To account for potential phylogenomic biases, we implemented various data extraction, locus sampling, and signal filtering strategies to generate matrices. Subsequently, we applied a diverse array of tree-searching and rate-modelling methods to reconstruct the phylogeny. Our analyses, utilizing different matrices and methods, converged on the same unrooted relationships among collembolan ingroups, supporting the current ordinal classification and challenging the monophyly of Arthropleona and Symphypleona s.l. However, discrepancies across analyses existed in the root of Collembola. Among various root positions, those based on more informative matrices and biologically realistic models, favoring a basal topology of Entomobryomorpha + (Symphypleona s.s. + (Neelipleona + Poduromorpha)), were supported by subsequent methodological assessment, topology tests, and rooting analyses. This optimal topology suggests multiple independent reduction of the pronotum in non-poduromorph orders and aligns with the plesiomorphic status of neuroendocrine organs and epicuticular structure of Entomobryomorpha. Fossil-calibrated dating analyses based on the optimal topology indicated late-Paleozoic to mid-Mesozoic origins of the crown Collembola and four orders. In addition, our results questioned the monophyly of Isotomidae and Neanuridae, underscoring the need for further attention to the systematics of these families. Overall, this study provides novel insights into the phylogenetic backbone of Collembola, which will inform future studies on the systematics, ecology, and evolution of this significant arthropod lineage.
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The poor durability of Ru-based catalysts limits the practical application in proton exchange membrane water electrolysis (PEMWE). Here, we report that the asymmetric active units in Ru1-xMxO2 (M = Sb, In, and Sn) binary solid solution oxides are constructed by introducing acid-resistant p-block metal sites, breaking the activity and stability limitations of RuO2 in acidic oxygen evolution reaction (OER). Constructing highly asymmetric Ru-O-Sb units with a strong electron delocalization effect significantly shortens the spatial distance between Ru and Sb sites, improving the bonding strength of the overall structure. The unique two-electron redox couples at Sb sites in asymmetric active units trigger additional chemical steps at different OER stages, facilitating continuous proton transfer. The optimized Ru0.8Sb0.2O2 solid solution requires a superlow overpotential of 160 mV at 10 mA cm-2 and a record-breaking stability of 1100 h in an acidic electrolyte. Notably, the scale-prepared Ru0.8Sb0.2O2 achieves efficient PEMWE performance under industrial conditions. General mechanism analysis shows that the enhanced proton transport in the asymmetric Ru-O-M unit provides a new working pathway for acidic OER, breaking the scaling relationship without sacrificing stability.
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To explore the characteristics of hematologic indicators and related risk factors of lower extremity deep vein thrombosis (LDVT) in patients with cerebral infarction. METHODS: This study retrospectively analyzed data from 174 patients with cerebral infarction admitted to The Rehabilitation Department of Shanghai Fifth Rehabilitation Hospital and Shanghai First People's Hospital from June 2022 to June 2023. Based on the results of lower limb venous color Doppler ultrasound examinations, patients were divided into two groups: the LDVT group (35 cases) and the non-LDVT group (139 cases). We compared the clinical data and hematologic indicators (D-dimer value, fibrinogen, white blood cells, platelets, uric acid, creatinine, etc.) of the two groups to identify the risk factors of cerebral infarction complicated with LDVT. RESULTS: Statistical analysis revealed that the D-dimer values of the LDVT group were significantly (P<0.05) higher than those of the non-LDVT group. The uric acid value of the LDVT group was significantly lower than that of the non-LDVT group, with statistical significance (P<0.05). The Brunnstrom staging in the LDVT group was significantly different from that in the non-LDVT group (P<0.05). Meanwhile, binary logistic regression analysis showed that LDVT complicated with cerebral infarction was associated with D-dimer level [OR=1.302, 95% CI (1.077, 1.575)], uric acid level [OR=0.995, 95% CI (0.990, 1.000)], and Brunnstrom staging [OR=3.005, 95% CI (1.312, 6.880)]. CONCLUSION: D-dimer value, uric acid value, and Brunnstrom stage I to II are closely related to the occurrence of LDVT in patients with cerebral infarction. High D-dimer value, low uric acid value, and Brunnstrom stage I to II are independent risk factors for LDVT in cerebral infarction. Early assessment of D-dimer value, uric acid value, and Brunnstrom stage of cerebral infarction should be considered in clinical practice.
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Collaborative management of environmental pollution and carbon emissions (CMPC) has been a major policy instrument to promote Sustainable Development Goals (SDG) in recent years. However, the relationship between the benefits and drawbacks of this environmental management practice for green growth in and around a local area remains to be clarified. Using 30 provinces in China during 2001-2019 as the object of analysis, we assessed the efficiency of local CMPC practices using the nonradial directional distance function (NDDF) model, predicted local green growth using the frontier green complexity index (GCI), and empirically examined the spatial effects, locational heterogeneity, and threshold characteristics of the relationship using the spatial Durbin model and the panel threshold model. Our study finds that although efficient CMPC does drive local green growth, the promotion effect is nonlinear with decreasing marginal effect. This effect is particularly obvious in economically developed regions with higher CMPCs, which will absorb resources from neighboring regions and create a "siphoning" effect. It was found that local financial support and foreign direct investment (FDI) can radiate green growth to neighboring regions; therefore, CMPC practice needs to pay more attention to the effect of joint governance, supplemented by financial and foreign investment policy tools, to better promote the green transformation of local economy.
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Background: There is little research on the relationship between flavonol consumption and chronic kidney disease (CKD). This study aimed to examine the link between flavonol consumption and the risk of CKD among US adults, using data from the 2007-2008, 2009-2010 and 2017-2018 National Health and Nutrition Examination Survey (NHANES). Methods: A cross-sectional approach was used, drawing on data from three NHANES cycles. The flavonol consumption of the participants in this study was assessed using a 48 h dietary recall interview. CKD was diagnosed based on an estimated glomerular filtration rate below 60 mL/min/1.73 m2 or a urine albumin-to-creatinine ratio of 30 mg/g or higher. Results: Compared to the lowest quartile of flavonol intake (Q1), the odds ratios for CKD were 0.598 (95% CI: 0.349, 1.023) for the second quartile (Q2), 0.679 (95% CI: 0.404, 1.142) for the third quartile (Q3), and 0.628 (95% CI: 0.395, 0.998) for the fourth quartile (Q4), with a p value for trend significance of 0.190. In addition, there was a significant trend in CKD risk with isorhamnetin intake, with the odds ratios for CKD decreasing to 0.860 (95% CI: 0.546, 1.354) in the second quartile, 0.778 (95% CI: 0.515, 1.177) in the third quartile, and 0.637 (95% CI: 0.515, 1.177) in the fourth quartile (p for trend = 0.013). Conclusion: Our analysis of the NHANES data spanning 2007-2008, 2009-2010, and 2017-2018 suggests that high consumption of dietary flavonol, especially isorhamnetin, might be linked to a lower risk of CKD in US adults. These findings offer new avenues for exploring strategies for managing CKD.
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High-concentration ozone pollution pose threats to ecosystems and human health. However, there is limited research on the impact of the alternating evolution of synoptic weather patterns (SWPs) on the multi-scale transport processes and sources of ozone. From June 14 to 18, 2018, a rare consecutive ozone pollution plagued in Hefei and broader Yangtze River Delta region (YRD). This study investigates the meteorological factors and sources using in-situ observational data and WRF-Chem model simulations. Analysis reveals a northeastern low-pressure system moving from north to south generated a cold front. This moving cold front facilitated the vertical transport of warm air masses carrying high-concentration ozone originating from North China. Subsequently, Ozone-rich air masses (ORMs) were transported over the YRD, influenced by the eastward movement of the Mongolian high-pressure system. Based on WRF-Chem model with NOx tagging mechanisms and WRF-FLEXPART backward simulations, it is confirmed that a notable atmospheric transport originated from North China region (NCR) to Hefei, especially on June 15. As the Mongolian high-pressure weakens and shifts east-southward, it carried ORMs generated by NOx emissions from the YRD, accumulating over the sea within the range of 120°E to 126°E and 25°N to 30°N. Both WRF-chem model results and TRopospheric Ozone and Precursors from Earth System Sounding (TROPESS) Chemistry Reanalysis dataset Version 2 (TCR-2) revealed the existence of ORMs in this geographic range. Subsequently, the ORMs carried out to sea by the weakened high-pressure system were reintroduced inland, influenced by southeast winds brought about by the peripheral circulation of typhoon "Gaemi". In summary, the alternating evolution of SWPs significantly influences multi-scale ozone transport from both the NCR and the YRD regions, making substantial contributions to this prolonged episode. These findings offer valuable insights for improving regional ozone pollution prevention and control mechanisms.
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Due to the stability issue, It is difficult to prepare a silver nanocluster bearing functional sites, especially at a large scale. We report the synthesis and structure of a stable silver nanocluster bearing multiple surface aldehyde groups [Ag21(Ph2PO2)10(p-CHOPhC≡C)6]SbF6, which allows for postsynthesis modification such as surface functionalization through aldimine condensation to give homochiral clusters. Remarkably, the preparation of this cluster can be done in ~90 % high yield at gram scale, which facilitates further studies and potential applications. Through DFT calculations and geometric structure analysis, the high stability of this cluster is attributed to the geometric closure and electronic structure. This is the first time that an effective one-pot method has been developed to synthesize functional silver nanoclusters in high yield. The title cluster will be useful in the development of a variety of cluster-based materials.
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Interleukin-17A therapeutic inhibitors are among the most effective treatment methods for moderate-to-severe plaque psoriasis (PP). Reflectance confocal microscopy is a non-invasive imaging technique already documented to be beneficial in evaluating the follow-up of PP under treatment with topical actives and phototherapy. This study aimed to assess the epidermal and dermal changes associated with psoriasis and its treatment with RCM during systemic secukinumab treatment in patients with moderate-to-severe PP. A pilot study was conducted to evaluate RCM as a non-invasive tool for monitoring secukinumab treatment in patients with PP. For patients receiving secukinumab treatment, lesional skin was selected for RCM imaging, which were recorded at all scheduled times. The RCM evaluation criteria were established based on the histopathological diagnostic criteria for psoriasis. The clinical severity of psoriasis was assessed utilizing the psoriasis area severity index. A total of 23 patients with PP were included in the study. Each patient received 300 mg of subcutaneous secukinumab as induction therapy at baseline and weeks 1-4, followed by maintenance therapy every four weeks. Microscopic confocal changes were observed during the treatment. The results identified early microscopic evidence of the anti-inflammatory activity of secukinumab, which was not detected during the clinical examination. RCM findings correlating with the PASI were used to observe the patient's response to treatment and were identified as follows: acanthosis and parakeratosis, presence of epidermal and dermal inflammatory cells, presence of non-edge dermal papillae, and vascularization in the papillary dermis. This study is the first to demonstrate the use of RCM as an effective tool for non-invasive monitoring of secukinumab therapeutic response at a cellular level in a clinical or research setting. Early detection of RCM parameters associated with secukinumab activity may facilitate the identification of an early treatment response. RCM appears to be capable of providing practical and helpful information regarding follow-up in patients with PP undergoing secukinumab treatment. RCM may also provide novel perspectives on the subclinical evaluation of PP's response to biological therapy.
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Anticuerpos Monoclonales Humanizados , Interleucina-17 , Microscopía Confocal , Psoriasis , Humanos , Psoriasis/tratamiento farmacológico , Psoriasis/diagnóstico por imagen , Psoriasis/patología , Interleucina-17/antagonistas & inhibidores , Microscopía Confocal/métodos , Femenino , Masculino , Anticuerpos Monoclonales Humanizados/uso terapéutico , Persona de Mediana Edad , Adulto , Proyectos Piloto , Estudios de Seguimiento , Anciano , Piel/patología , Piel/diagnóstico por imagen , Resultado del Tratamiento , Índice de Severidad de la Enfermedad , Anticuerpos Monoclonales/uso terapéuticoRESUMEN
BACKGROUND: Management of inferior ramus of the pubis-ischium ramus remains controversial, and related research is sparse. The main intention of this study is to describe the biomechanical and clinical outcomes of pubis-ischium ramus fractures in Tile B pelvic injuries and to identify the feasibility and necessity of fixation of the inferior ramus of the pubis-ischium ramus. METHODS: This study comprised two parts: a biomechanical test and a retrospective clinical study. For the biomechanical tests, Tile B-type pelvic injuries were modeled in six cadaver specimens by performing pubis-ischium osteotomies and disruption of the anterior and interosseous sacroiliac ligaments. The superior and/or inferior rami of the pubis-ischium ramus were repaired with reconstruction plates and separated into three groups (A, B, and C). Specimens were placed in the standing position and were loaded axially with two-leg support for three cycles at 500 N. The displacements of sacroiliac joints at osteotomy were measured with Vernier calipers and compared using statistical software. To investigate the clinical outcomes of this technique, 26 patients were retrospectively analyzed and divided into a superior ramus fixation group (Group D) and a combined superior and inferior ramus of the pubis-ischium ramus fixation group (Group E). The main outcome measures were time of operation, blood loss, postoperative radiographic reduction grading, and functional outcomes. RESULTS: In the vertical loading test, Group E showed better pelvic ring stability than Group D (P < 0.05). However, the shift of the sacroiliac joints was almost identical among the three groups. In our clinical case series, all fractures in Group E achieved bony union. Group E demonstrated earlier weight-bearing functional exercise (2.54 ± 1.45 vs 4.77 ± 2.09; P = 0.004), earlier bony union (13.23 ± 2.89 vs 16.55 ± 3.11; P = 0.013), and better functional outcomes (89.77 ± 7.27 vs 82.38 ± 8.81; P = 0.028) than Group D. The incidence of sexual dysfunction was significantly lower in Group E than that in Group D (2/13 vs 7/13; P = 0.039). Bone nonunion occurred in two patients in Group D, and two patients in Group E had heterotopic ossification. None of the patients exhibited wound complications, infections, implant failures, or bone-implant interface failures. CONCLUSIONS: Fixation of the inferior ramus of a pubis-ischium ramus fracture based on conventional fixation of the anterior pelvic ring is mechanically superior in cadaveric Tile B pelvic injury and shows rapid recovery, good functional outcomes, and low incidence of complications.
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Placas Óseas , Huesos Pélvicos , Humanos , Fenómenos Biomecánicos , Masculino , Femenino , Adulto , Huesos Pélvicos/cirugía , Huesos Pélvicos/lesiones , Huesos Pélvicos/diagnóstico por imagen , Persona de Mediana Edad , Fenómenos Mecánicos , Cadáver , Fracturas Óseas/cirugía , Estudios Retrospectivos , Fijación Interna de Fracturas/instrumentaciónRESUMEN
The desire to explore the natural resources and geopolitical patterns of the North and South Poles has significantly increased the interest of experts and researchers in the development and utilization of the polar regions. In this article, we comprehensively analyzed the current state of the development of polar low-temperature steel around the world. We highlighted the challenges that must be addressed in the ongoing development efforts and summarized the expected future trends in this field. The main theme of this article involves the challenges encountered in polar environments primarily caused by the low-temperature toughness and seawater corrosion of marine steel.
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Constructed wetlands (CWs) have been widely used to ensure effective domestic wastewater treatment. Microorganisms-derived CWs have received extensive attention as they play a crucial role. However, research on the succession patterns of microbial communities and the influencing mechanisms of internal environmental factors throughout entire CW operations remains limited. In this context, three parallel-operated CWs were established in this study to assess the microbial communities and their influencing environmental factors at different substrate depths throughout the operation process using 16S rRNA gene high-throughput sequencing and metagenomic sequencing. The results showed gradual reproduction and accumulation of the microbial communities throughout the CW operation. Although gradual increases in the richness and diversity of the microbial communities were found, there were decreases in the functional expression of the dominant microbial species. The excessive accumulation of microorganisms will decrease the oxidation-reduction potential (ORP) within CWs and attenuate their influence on effluent. Dissolved oxygen (DO) was the major factor influencing the microbial community succession over the CW operation. The main identified functional bacterial genera responsible for the ammonium oxidation, nitrification, and denitrification processes in the CWs were Nitrosospira, Nitrobacter, Nitrospira, Rhodanobacter, and Nakamurella. The narG gene was identified as a key functional gene linking various components of nitrogen cycling, while pH, electrical conductivity (EC), and ORP were the major environmental factors affecting the metabolism characteristics of nitrogen functional microorganisms. This study provides a theoretical basis for the effective regulation of related microbial communities to achieve long-term, efficient, and stable CW operations.
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Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA) are the four main pathogens that cause viral diarrhea in pigs, and they often occur in mixed infections, which are difficult to distinguish only according to clinical symptoms. Here, we developed a multiplex TaqMan-probe-based real-time RT-PCR method for the simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA for the first time. The specific primers and probes were designed for the M protein gene of PEDV, N protein gene of TGEV, N protein gene of PDCoV, and VP7 protein gene of PoRVA, and corresponding recombinant plasmids were constructed. The method showed extreme specificity, high sensitivity, and excellent repeatability; the limit of detection (LOD) can reach as low as 2.18 × 102 copies/µL in multiplex real-time RT-PCR assay. A total of 97 clinical samples were used to compare the results of the conventional reverse transcription PCR (RT-PCR) and this multiplex real-time RT-PCR for PEDV, TGEV, PDCoV, and PoRVA detection, and the results were 100% consistent. Subsequently, five randomly selected clinical samples that tested positive were sent for DNA sequencing verification, and the sequencing results showed consistency with the detection results of the conventional RT-PCR and our developed method in this study. In summary, this study developed a multiplex real-time RT-PCR method for simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA, and the results of this study can provide technical means for the differential diagnosis and epidemiological investigation of these four porcine viral diarrheic diseases.