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ABSTRACT: This study aimed to investigate the effects of dietary Bacillus velezensis KNF-209 (BV-KNF-209) on the growth performance, immunity, and gut health of broilers. A total of 540 one-day-old male Cobb-500 broilers were randomly divided into 5 groups of 6 replicates with 18 broilers per replicate. Dietary treatments were corn-soybean meal basal diets supplemented with 0, 50, 100, 200, and 400 mg/kg BV-KNF-209 (CON, BV 50, BV 100, BV 200, and BV 400 groups, respectively) for 42 d. Compared with the CON group, the average daily gains (ADG) at 0 to 42 d in the BV 100 and BV 200 groups were significantly increased (P < 0.01), and the feed-to-gain (F:G) ratios were significantly decreased at 0 to 21 d (P < 0.01) and 0 to 42 d (P < 0.05). The BV 200 and BV 400 groups had higher serum immunoglobulin M (IgM) levels at d 21 and 42 (P < 0.05). The serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were significantly decreased in the BV 50, BV 100, and BV 200 groups at d 21 (P < 0.05), and serum IL-1β and IL-6 levels were also reduced in the BV 100 and BV 200 groups at d 42 (P < 0.05). Meanwhile, increased interleukin-10 (IL-10) levels in the jejunal and ileal mucosa at d 42 were observed in the BV 100, BV 200, and BV 400 groups (P < 0.05), while the IL-1β and IL-6 levels (P < 0.01) were decreased. The BV 200 and BV 400 groups showed significantly higher activities of lipase and trypsin (P < 0.05) in jejunal digesta as well as higher activities of amylase and trypsin (P < 0.01) in ileal digesta at d 42. The cecal acetic acid and propionic acid levels in the BV groups and lactic acid levels in the BV 50, BV 100, and BV 200 groups (P < 0.05) were significantly higher compared to those in the CON group. Overall, dietary BV-KNF-209 supplementation significantly improved broiler growth performance, an effect that may have been achieved by heightening immunity, increasing digestive enzyme activity, and raising intestinal short-chain fatty acids and lactic acid levels.

The expansion of hydraulic fractures in coalbed methane (CBM) reservoirs is key to effective stimulation, making it essential to understand fracture propagation and its influencing factors for efficient resource development. Using petrological characteristics, logging data, microseismic monitoring, and fracturing reports from the Baode Block on the eastern Ordos Basin, this study systematically investigates the geological and engineering factors influencing hydraulic fracture propagation. The real-time monitoring of fracture propagation in 12 fractured wells was conducted using microseismic monitoring techniques. The results indicated that the fracture orientations in the study area ranged from NE30° to NE60°, with fracture lengths varying between 136 and 226 m and fracture heights ranging from 8.5 to 25.3 m. Additionally, the fracturing curves in the study area can be classified into four types: stable, descending, fluctuating, and falling. Among these, the stable and descending types exhibit the most effective fracture propagation and are more likely to generate longer fractures. In undeformed–cataclastic coals and bright and semi-bright coals, long fractures are likely to form. When the Geological Strength Index (GSI) of the coal rock ranges between 60 and 70, fracture lengths generally exceed 200 m. When the coal macrolithotype index (Sm) is below 2, fracture lengths typically exceed 200 m. When the difference between the maximum and minimum horizontal principal stresses exceeds 5 MPa, fractures with length >180 m are formed, while fracture heights generally remain below 15 m. From an engineering perspective, for the study area, hydraulic fracturing measures with a preflush ratio of 20–30%, an average sand ratio of 13–15%, and a construction pressure between 15 MPa and 25 MPa are most favorable for coalbed methane production.

Marine shale reservoirs have experienced intense tectonic movements in southeastern Chongqing, resulting in widely developed folds, faults and bedding slip deformation. The gas content is of a wide range while the organic matter content and thickness are sufficient for shale gas accumulation. Here, we review previous work and present outcrop and core data from 11 shale wells around southeastern Chongqing to analyze and summarize tectonic types and their combinations, as well as their influence on shale gas preservation and enrichment. Fault and bedding slips are prevalent in shale reservoirs, and they apparently influence shale gas content. Bedding slip crush is usually formed at the bottom interval where the total organic content is high from intense horizontal compression. Bedding slip crushing and well-developed high-angle natural fractures form a net fracture system where shale gas is enriched in an anticline structure or lost to air in a syncline structure laterally. Strata dip is the key geological factor that controls bedding slip deformation in shale reservoirs in certain areas. According to the established bedding slip geological model, when bedding slip occurred, the critical strata dips of the lower Silurian Longmaxi Formation were 13.34°, respectively. Bedding slip crushed anticlines without faults and synclines without faults and bedding slip crushing are the two tectonic patterns that are favorable for shale gas preservation and enrichment, which should be the focus of further explorations of lower Paleozoic shale gas in southeastern Chongqing.

Erlian Basin is a Mesozoic oil and gas-bearing basin in northeast of China. The extension rate, extension direction, and stratum thickness of this rift basin have a clear control on its shape and extension. In this study, we design three sandbox models of the Erlian Basin to represent the effects of changing these three factors. The extension rate controls the timing of secondary fault formation inside the rift basin; a high extension rate promotes faster deformation inside the rift. The extension direction controls the strike of the fault inside the rift; a greater angle between the extension direction and the normal direction of the strike of the rift favors rapid evolution of internal secondary faults. The stratum thickness represents the control of sedimentation on the rift basin; the thinner the brittle layer, the wider the rift. The simulation results also show that the extension direction is the major factor controlling tectonic deformation in the basin. Stratum thickness and extension rate are secondary controlling factors. Additionally, according to geometric and kinematic similarities between typical Mesozoic rift basins in eastern and adjacent areas of China, we suggest that southeastward extension is a possible kinematic mechanism for basin formation.

This study was conducted to investigate the effects of 1,8-cineole on antioxidant capacity, immunity, and intestinal health of broilers. A total of 540 1-day-old Arbor Acres (AA) male broilers were randomly divided into five treatments with six replicates per treatment, and 18 broilers per replicate for 42 days. Dietary treatments were a corn–soybean meal basal diet supplemented with 0, and 10, 20, 30, and 40 mg/kg 1,8-cineole, respectively. Dietary supplementation with 20~30 mg/kg of 1,8-cineole increased the ADG from d 22 to 42 and d 1 to 42 (p < 0.05), and decreased the FCR (p < 0.05). Dietary supplementation of 10~40 mg/kg of 1,8-cineole increased total antioxidant capacity (TAOC) in serum (p < 0.05), and decreased malondialdehyde (MDA) level in the liver on day 21 (p < 0.05). The supplementation of 20~30 mg/kg of 1,8-cineole increased the activity of total superoxide dismutase (T-SOD) in the serum and liver and TAOC in the serum and the liver (p < 0.05), and decreased the level of MDA in the serum and the liver (p < 0.05) on day 42. Dietary supplementation with 20~30 mg/kg of 1,8-cineole increased serum immunoglobulin A, immunoglobulin G, and immunoglobulin M contents on day 21 (p < 0.05). On day 21, dietary supplementation of 20~30 mg/kg of 1,8-cineole increased the VH and VH/CD (p < 0.05) in the jejunum and ileum. The supplementation of 20~30 mg/kg of 1,8-cineole increased the content of secretory immunoglobulin A in the duodenum and ileum mucosa on d 42 (p < 0.05). In conclusion, dietary supplementation of 1,8-cineole improves the growth performance of broilers by enhancing antioxidant capacity, immunity, and intestinal morphology.

Carbendazim (CAR) and enrofloxacin (ENF) are frequently detected in fruits and meat products, respectively. Since most people consume fruits, vegetables, and meat products, combined exposure is possible, necessitating further evaluation of toxic interactions. In this study, the developmental toxicity of separate and combined exposure was examined in zebrafish embryos. Carbendazim exposure at 0.79 mg/L and above significantly affected developmental parameters, while enrofloxacin alone had no substantial effects on these developmental parameters within the selected concentration range (0.10–0.40 mg/L). Surprisingly, ENF antagonized the CAR-evoked reduction in the 48 hpf (hours post-fertilization) hatching rate and the increases in the 96 hpf malformation and lethality rates. The results revealed that the antagonism might be associated with reciprocal effects of these compounds on metabolism-related genes, such as cyp7a1 and apoa1a. These results reveal a complex interaction between ENF and CAR on metabolic regulation during development and highlight the importance of combined assessment for agents with the potential for simultaneous exposure.

Abstract Background RNA methylation, an important reversible post-transcriptional modification in eukaryotes, has emerged as a prevalent epigenetic alteration. However, the role of the m6A reader YTH domain family 2 (YTHDF2) has not been reported in anaplastic thyroid cancer (ATC) and its biological mechanism is unclear. Methods The relationship between YTHDF2 expression and ATC was determined using data sets and tissue samples. A range of analytical techniques were employed to investigate the regulatory mechanism of YTHDF2 in ATC, including bioinformatics analysis, m6A dot-blot analysis, methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNA immunoprecipitation (RIP) assays, RNA sequencing, RNA stability assays and dual luciferase reporter gene assays. In vitro and in vivo assays were also conducted to determine the contribution of YTHDF2 to ATC development. Results YTHDF2 expression was significantly increased in ATC. The comprehensive in vitro and in vivo experiments demonstrated that YTHDF2 knockdown significantly attenuated ATC proliferation, invasion, migration, and apoptosis promotion, whereas YTHDF2 overexpression yielded the opposite trend. Mechanistically, RNA-seq, MeRIP-seq and RIP-seq analysis, and molecular biology experiments demonstrated that YTHDF2 accelerated the degradation of DNA damage-inducible transcript 4 or regulated in DNA damage and development 1 (DDIT4, or REDD1) mRNA in an m6A-dependent manner, which in turn activated the AKT/mTOR signaling pathway and induced activation of epithelial-mesenchymal transition (EMT), thereby promoting ATC tumor progression. Conclusions This study is the first to demonstrate that elevated YTHDF2 expression levels suppress DDIT4 expression in an m6A-dependent manner and activate the AKT/mTOR signaling pathway, thereby promoting ATC progression. YTHDF2 plays a pivotal role in ATC progression, and it may serve as a promising therapeutic target in the future.

Abstract The outbreaks of severe acute respiratory syndrome coronavirus (SARS-CoV-1), Middle East respiratory syndrome coronavirus (MERS-CoV), and SARS-CoV-2 highlight the need for countermeasures to prevent future coronavirus pandemics. Given the unpredictable nature of spillover events, preparing antibodies with broad coronavirus-neutralizing activity is an ideal proactive strategy. Here, we investigated whether SARS-CoV-2 infection and vaccination could provide cross-neutralizing antibodies (nAbs) against zoonotic sarbecoviruses. We evaluated the cross-neutralizing profiles of plasma and monoclonal antibodies constructed from B cells from coronavirus disease 2019 (COVID-19) convalescents and vaccine recipients; against sarbecoviruses originating from bats, civets, and pangolins; and against SARS-CoV-1 and SARS-CoV-2. We found that the majority of individuals with natural infection and vaccination elicited broad nAb responses to most tested sarbecoviruses, particularly to clade 1b viruses, but exhibited very low cross-neutralization to SARS-CoV-1 in both natural infection and vaccination, and vaccination boosters significantly augmented the magnitude and breadth of nAbs to sarbecoviruses. Of the nAbs, several exhibited neutralization activity against multiple sarbecoviruses by targeting the spike receptor-binding domain (RBD) and competing with angiotensin-converting enzyme 2 (ACE2) binding. SCM12-61 demonstrated exceptional potency, with half-maximal inhibitory concentration (IC50) values of 0.001–0.091 μg/mL against tested sarbecoviruses; while VSM9-12 exhibited remarkable cross-neutralizing breadth against sarbecoviruses and SARS-CoV-2 Omicron subvariants, highlighting the potential of these two nAbs in combating sarbecoviruses and SARS-CoV-2 Omicron subvariants. Collectively, our findings suggest that vaccination with an ancestral SARS-CoV-2 vaccine, in combination with broad nAbs against sarbecoviruses, may provide a countermeasure for preventing further sarbecovirus outbreaks in humans.

Abstract Long-term humoral immunity to SARS-CoV-2 is essential for preventing reinfection. The production of neutralizing antibody (nAb) and B cell differentiation are tightly regulated by T follicular help (TFH) cells. However, the longevity and functional role of TFH cell subsets in COVID-19 convalescents and vaccine recipients remain poorly defined. Here, we show that SARS-CoV-2 infection and inactivated vaccine elicited both spike-specific CXCR3+ TFH cell and CXCR3− TFH cell responses, which showed distinct response patterns. Spike-specific CXCR3+ TFH cells exhibit a dominant and more durable response than CXCR3− TFH cells that positively correlated with antibody responses. A third booster dose preferentially expands the spike-specific CXCR3+ TFH cell subset induced by two doses of inactivated vaccine, contributing to antibody maturation and potency. Functionally, spike-specific CXCR3+ TFH cells have a greater ability to induce spike-specific antibody secreting cells (ASCs) differentiation compared to spike-specific CXCR3− TFH cells. In conclusion, the persistent and functional role of spike-specific CXCR3+ TFH cells following SARS-CoV-2 infection and vaccination may play an important role in antibody maintenance and recall response, thereby conferring long-term protection. The findings from this study will inform the development of SARS-CoV-2 vaccines aiming to induce long-term protective immune memory.

ABSTRACT Dengue virus (DENV) is the most widespread arbovirus, causing symptoms ranging from dengue fever to severe dengue, including hemorrhagic fever and shock syndrome. Four serotypes of DENV (DENV-1 to -4) can infect humans; however, no anti-DENV drug is available. To facilitate the study of antivirals and viral pathogenesis, here we developed an infectious clone and a subgenomic replicon of DENV-3 strains for anti-DENV drug discovery by screening a synthetic compound library. The viral cDNA was amplified from a serum sample from a DENV-3-infected individual during the 2019 epidemic; however, fragments containing the prM-E-partial NS1 region could not be cloned until a DENV-3 consensus sequence with 19 synonymous substitutions was introduced to reduce putative Escherichia coli promoter activity. Transfection of the resulting cDNA clone, plasmid DV3syn, released an infectious virus titer of 2.2 × 102 focus-forming units (FFU)/mL. Through serial passages, four adaptive mutations (4M) were identified, and addition of 4M generated recombinant DV3syn_4M, which produced viral titers ranging from 1.5 × 104 to 6.7 × 104 FFU/mL and remained genetically stable in transformant bacteria. Additionally, we constructed a DENV-3 subgenomic replicon and screened an arylnaphthalene lignan library, from which C169-P1 was identified as exhibiting inhibitory effects on viral replicon. A time-of-drug addition assay revealed that C169-P1 also impeded the internalization process of cell entry. Furthermore, we demonstrated that C169-P1 inhibited the infectivity of DV3syn_4M, as well as DENV-1, DENV-2, and DENV-4, in a dose-dependent manner. This study provides an infectious clone and a replicon for the study of DENV-3 and a candidate compound for future development against DENV-1 to -4 infections. IMPORTANCE Dengue virus (DENV) is the most prevalent mosquito-transmitted virus, and there is no an anti-dengue drug. Reverse genetic systems representative of different serotype viruses are invaluable tools for the study of viral pathogenesis and antiviral drugs. Here, we developed an efficient infectious clone of a clinical DENV-3 genotype III isolate. We successfully overcame the instability of flavivirus genome-length cDNA in transformant bacteria, an unsolved issue for construction of cDNA clones of flaviviruses, and adapted this clone to efficiently produce infectious viruses following plasmid transfection of cell culture. Moreover, we constructed a DENV-3 subgenomic replicon and screened a compound library. An arylnaphthalene lignan, C169-P1, was identified as an inhibitor of virus replication and cell entry. Finally, we demonstrated that C169-P1 exhibited a broad-spectrum antiviral effect against the infections with DENV-1 to -4. The reverse genetic systems and the compound candidate described here facilitate the study of DENV and related RNA viruses.

Soft rock has the characteristics of poor engineering mechanics, and it is easy to change under the action of water or external load. This paper uses the pulse attenuation permeability meter to develop permeability test and coefficient of variation for weak rock with common engineering mechanical properties. Calculate and explore the geological stress sensitivity characteristics of weak rock engineering within the Three Goiges reseijvoir area. The conclusions are as follows: the permeability of weak rock in the Three Gorges reservoir area is less than 0.lmD, which belongs to the type of compact soft rock, among which the permeability of medium sandstone (0.048mD) bigger than the permeability of fine sandstone (0.0055mD), fine sandstone bigger than the permeability of mud shale (0.0047mD); the rock permeability coefficient of variation is greater than 0.7, with a strong heterogeneity. The permeability of soft rock has obvious anisotropy. The permeability of the vertical layer of medium-fine sandstone and silty mudstone is greater than the permeability parallel to the plane. The difference between the two is closely related to the physical difference between different lithologies. The permeability of siltstone, shale and sandstone rocks decreases exponentially with the increase of effective stress. The relationship between the permeability loss rates of various types of rock is: shale bigger than siltstone, siltstone bigger than sandstone, indicating the same stress condition, page Rocks have higher stress sensitivity than siltstones and sandstones. This is mainly because the fractures in sandstone and siltstone are more developed than shale. [ABSTRACT FROM AUTHOR]

Two-dimensional (2D) Ti3C2Tx MXene sheet possesses its interesting electrical and optical properties and has thus aroused widespread interest in a wide range of flexible optoelectronic device applications. A clear understanding of the friction behavior and performance, of MXene and other transparent materials (e.g., SiO2) will enable these promising applications. In this study, the frictional behaviors between the SiO2 (100) plane and the MXene sheet have been measured by a lateral force atomic force microscopy (AFM), based on the realization of the friction experiments in the single crystal heterojunction. The results on the two-dimensional single–layer interface between MXene and SiO2 clearly show that the corresponding heterostructure owns a strong friction anisotropy with a six-fold symmetry. The friction coefficient for the misaligned contact is 0.067, while it is 0.031 for the aligned one. We find that the measurement of friction anisotropy is highly related to the number of atomic layers of MXene. The increase of the layer number is found to significantly reduce the friction anisotropy and beyon 10 layers it is no longer related to the thickness. These findings will have general applicability and can be used in other MXene heterojunction configurations to realize the advantages of adaptive nanodevices and the development of flexible optoelectronics.

BackgroundPrediction of central lymph node metastasis (CLNM) is vital for clinical decision-making processes in clinically N0 (cN0) unifocal papillary thyroid carcinoma (PTC), but the sensitivity of preoperative detection of CLNM is limited. The aim of the present study was to determine whether there are ultrasonic (US) characteristics associated with CLNM.MethodsIn total, 1657 PTC patients (514 men and 1143 women) were enrolled in the present study between January 2018 and May 2021. The patients met the following inclusion criteria based on preoperative detection: suspected nodule confirmed as PTC by biopsy; the nodule was unifocal and less than 4 cm in diameter; no prior neck radiation exposure; no extrathyroidal extension; and no CLNM or distant metastases on imaging. All the enrolled patients underwent total thyroidectomy with prophylactic central lymph node dissection (CLND). A postoperative pathological diagnosis was made.ResultsCLNM was found in 58.4% of male patients and 36.9% of female patients. In univariate analysis, size, adjacent anterior capsule, distance to the lower pole and color Doppler flow imaging (CDFI) were considered risk factors for the male and female groups (p < 0.05). In multivariate analyses, size, adjacent anterior capsule, distance to the lower pole and CDFI were independent risk factors for male patients. For females, the independent risk factors included size, adjacent anterior capsule, distance to the lower pole and CDFI.ConclusionIn the present cohort, US imaging characteristics, including size, adjacent anterior capsule, distance to the lower pole and CDFI, were identified to be potentially beneficial in preoperative clinical decision-making processes for cN0 unifocal PTC patients.

BackgroundThyroid nodules are a very common finding in the general population. Fine-needle aspiration (FNA) has been recommended as the initial test for the evaluation of thyroid nodules. The trend on reporting as atypia of undetermined significance (AUS) has been significantly increased, but the malignant risk is still controversial among different studies. The aim of this study is to investigate the malignancy risk of thyroid nodules reported as Bethesda category III (AUS/FLUS) on initial FNA.MethodWe reviewed 272 papillary thyroid cancer (PTC) patients with suspicious thyroid nodules who underwent fine-needle aspiration and received surgical treatment during 2019 to 2020.ResultsOne hundred ten (40.4%) patients were diagnosed with PTC. Multivariate analysis showed that microcalcification (p = 0.037, OR = 2.260, 95% CI: 1.051–4.860), shape (p = 0.003, OR = 4.367, 95% CI: 1.629–11.705), diameters (p = 0.002, OR = 0.278, 95% CI: 0.123–0.631), anti-thyroglobulin antibodies (TGAb) (p = 0.002, OR = 0.150, 95% CI: 0.046–0.494), anti-thyroid peroxidase antibody (A-TPO) (p = 0.009, OR = 4.784, 95% CI: 1.486–15.401), and nodule goiter (p < 0.001, OR = 0.100, 95% CI: 0.046–0.217) were independent malignant risk factors in patients with thyroid nodule classified as Bethesda category III.ConclusionIn this study, malignant risk factors in patients with thyroid nodule classified as Bethesda category III were significantly associated with preoperative serum TGAb, A-TPO, microcalcification, irregular shape, and nodule diameters. Nodules with malignant factors should be carefully elevated; surgery may be the better option for those patients.

Large‐volume central lymph node metastasis (large-volume CLNM) is associated with high recurrence rate in papillary thyroid carcinoma (PTC) patients. However, sensitivity in investigating large-volume CLNM on preoperative ultrasonography (US) is not high. The aim of this study is to investigate the clinical factors associated with large-volume CLNM in clinical N0 PTC patients. We reviewed 976 PTC patients undergoing total thyroidectomy with central lymph node dissection during 2017 to 2019. The rate of large-volume LNM was 4.1% (40 of 967 patients). Multivariate analysis showed that male gender and young age (age2cm) was associated with large-volume CLNM, with OR (95% CI) 3.757 (1.061–13.310), by multivariate analysis. In ROC curve analysis on the diameter of the CPTC tumor, the Area Under Curve (AUC) =0.682(p=0.013), the best cut-off point was selected as 2.0cm. In conclusion, male gender and young age were predictors for large-volume CLNM of cN0 PTC. cN0 PTMC patient with capsule invasion and cN0 CPTC patient with tumor diameter >2cm were correlated with large-volume CLNM. Total thyroidectomy with central lymph node dissection may be a favorable primary treatment option for those patients.

A dynamically adjustable terahertz (THz) broadband absorber based on a vanadium dioxide (VO2) hybrid metamaterial is theoretically designed. During the design, we first consider working frequency, bandwidth, excitation method and fabrication process. And then, a commercialized full wave electromagnetic (EM) calculation software CST microwave 2019 based on finite integration technology is used to simulate and optimize the performance of the device. Due to the device is excited by light, it consists of a top layer of VO2 oblique-split-ring-resonator (OSRR) array, a polyimide (PI) spacer and a bottom metal film. The simulation results show that, when the VO2 changes from insulating state to metal state under light stimulus, the absorption increases from 3% to over 90%. Meanwhile, when the VO2 is in metallic phase, the bandwidth of absorption greater than 90% reaches to 1.0 THz and the relative bandwidth is about 83%. The potential applications of the device are active EM stealth, THz wave detection and thermal imaging. This work paves a new way for simply constructing a low-cost dynamically adjustable THz broadband absorber.