Effects of Steviol Glycosides on Growth Performance, Ruminal Fermentation and Microbial Diversity of Hu Sheep.

The experiment was conducted to investigate the potential effects of steviol glycosides on growth performance, rumen fermentation processes, and microbial diversity in Hu sheep. A single-factor design was used for the trial. Twenty healthy weaned Hu lambs, possessing comparable body weights averaging 18.31 ± 1.24 kg, were randomly allocated into two distinct groups: the control group (CON) and the experimental group (STE), with each comprising 10 lambs. The CON was fed the basal diet, and the STE was supplemented with 0.07% steviol glycosides based on the basal diet. During the experimental period, variations in body weight and feed intake were closely monitored and recorded. After feeding for 90 d, blood was collected to determine blood biochemical indices, and rumen fluid samples were gathered for an in-depth analysis of rumen fermentation parameters and microbial diversity. The outcomes revealed no statistically significant differences in growth performance or serum biochemical indices between the two groups (p > 0.05). Rumen pH in STE and CON was within the normal range. The rumen ammonia nitrogen (NH3-N) and acetic acid (AA) content of STE decreased significantly compared with CON (p < 0.05). No significant variations were observed in the levels of other volatile fatty acids (VFAs) between the two groups (p > 0.05). The rumen microbial OTUs count, as well as the Shannon, Simpson, Chao1, and Ace indices, were notably lower in the STE group compared to the CON group (p < 0.05). Additionally, at the phylum level, the relative abundance of Firmicutes, Bacteroidetes, and Proteobacteria collectively accounted for over 97% of the total phylum composition. In comparison to the CON group, the STE group exhibited an increase in the relative abundance of Proteobacteria (p < 0.05), accompanied by a significant reduction in the relative abundance of Patescibacteria and Desulfobacteria (p < 0.05). At the genus level, there was a notable increase in the relative abundance of Prevotella_7 and Succinivibrionaceae_UCG_001 in the STE group, whereas the relative abundance of Rikenellaceae_RC9_gut_group significantly decreased (p < 0.05). According to the correlation analysis between rumen microflora and VFAs, the relative abundance of Succinivibrionaceae_UCG_001 displayed a significant negative correlation with AA (p < 0.05), whereas Lactobacillus exhibited a notable positive correlation with isobutyric acid (IBA) (p < 0.05). In summary, steviol glycosides had no significant effect on the production performance and blood biochemical indexes of Hu sheep. Steviol glycosides can improve rumen fermentation parameters and rumen microflora structure of Hu sheep and have a certain effect on rumen microbial diversity and composition.

Efficient degradation of tetracycline via peroxymonosulfate activation by phosphorus-doped biochar loaded with cobalt nanoparticles.

The accumulation of tetracycline hydrochloride (TCH) threatens human health because of its potential biological toxicity. Carbon -based materials with easy isolation and excellent performance that can activate peroxymonosulfate (PMS) to generate reactive oxygen species for TCH degradation are essential, but the development of such materials remains a significant challenge. In this study, based on the idea of treating waste, tricobalt tetraoxide loaded P-doped biochar (Co NP-PBC) was synthesised to activate PMS for the degradation of TCH. Possible degradation pathways and intermediate products of TCH were identified using High performance liquid chromatography tandem mass spectrometry (HPLC-MS) detection and density functional theory analysis. Toxicity analysis software was used to predict the toxicity of the intermediate products. Compared to catalysts loaded with Fe and Mn and other Co-based catalysts, Co NP-PBC exhibited an optimal performance (with a kinetic constant of 0.157 min-1 for TCH degradation), and over 99.0% of TCH can be degraded within 20 min. This mechanism demonstrates that the non-free radical oxidation of 1O2 plays a major role in the degradation of TCH. This study provides insights into the purification of wastewater using BC-based catalysts.

First report of leaf spot disease caused by Neopestalotiopsis rosae on Lonicera caerulea L. in Heilongjiang Province, China.

Recently, interest in cultivating blue honeysuckle (Lonicera caerulea L.) for horticulture and medicinal uses has grown (Sharma and Lee 2021). Between September 2022 and September 2023, a leaf spot disease (Fig. S1) was observed on approximately 20% of 'Lanjingling' blue honeysuckles grown in a 0.18 ha field in Qiqihar city (123.43°E, 47.92°N), Heilongjiang Province, China. Infected plants displayed black leaf spots that expanded to cover the entire leaf. Small, 3 to 4 mm segments of infected tissue were surface sterilized with 75% ethanol for 30 s and 5% sodium hypochlorite (NaOCl) for 3 min, rinsed three times with sterile distilled water, dried on paper towels, and plated in 9 cm Petri dishes containing potato dextrose agar (PDA) (Ma et al. 2023). To induce sporulation, nine purified cultures (Fig. S2) with similar culture characteristics were finally obtained from ten infected plants and they displayed a conidia morphology consistent with Neopestalotiopsis spp., no other fungi were isolated, and the isolation frequency was 90%. Conidiomata (Fig. S3) were brown to black and distributed in concentric rings with an average size of 261.98 (60.30-451.80) µm (n = 50). The conidia (Fig. S3) were fusoid and had four septa, straight to slightly curved, with an average size of 23.48 (13.50-30.30) × 5.42 (4.50-9.30) µm(n = 50), while basal and apical cells were hyaline and the three middle cells were brown with darker septa. PCR amplification was performed with ITS1/ITS4 (White et al. 1990), EFl-728F/EF1-986R (Carbone and Kohn 1999), and Btub2Fd/Btub4Rd (Glass and Donaldson 1995) primers from the genomic DNA of the LD-330. Sequences of ITS (PP033584), TEF (PP048757), and TUB (PP048758) revealed 99 to 100% (499/500, 255/255, and 481/486) shared identity with Neopestalotiopsis rosae sequences (NR145243, KM199524, and KM199430) (Rebollar-Alviter et al. 2020). Therefore, based on morphological characteristics and molecular phylogeny, LD-330 was identified as N. rosae. Six two-year-old healthy plants of the 'Lanjingling' cultivar were selected for a pathogenicity test (Yan et al. 2023). The leaves were surface disinfected with 75% alcohol and then wiped with sterilized water three times. Three plants were inoculated with 10 ml of LD-330 conidial suspension (1 × 106 spores/ml) or with sterile water as an experimental control, respectively. All plants were in closed plastic bag, incubated in a greenhouse at 28 ℃ and 75% relative humidity (RH) under a 12-h light/dark cycle, and each experiment was performed three times (Rebollar-Alviter et al. 2020). Typical leaf spot symptoms were observed on inoculated leaves after 14 days (Fig. S4), whereas no symptoms were detected on water-treated leaves. The same pathogen was reisolated from infected leaves, displayed the same morphological and molecular traits, and was again identified as N. rosae, confirming Koch's postulate. Neopestalotiopsis rosae was previously reported on pecan (Gao et al. 2022), causing black leaf spot disease in China. To our knowledge, this is the first report of a blue honeysuckle leaf spot caused by N. rosae in China and specifically in the Heilongjiang province which has the largest blue honeysuckle cultivation area in the country. Future research should be directed toward developing comprehensive management measures.

3D Printed Porous Zirconia Biomaterials based on Triply Periodic Minimal Surfaces Promote Osseointegration In Vitro by Regulating Osteoimmunomodulation and Osteo/Angiogenesis.

The triply periodic minimal surface (TPMS) is a highly useful structure for bone tissue engineering owing to its nearly nonexistent average surface curvature, high surface area-to-volume ratio, and exceptional mechanical energy absorption properties. However, limited literature is available regarding bionic zirconia implants using the TPMS structure for bone regeneration. Herein, we employed the digital light processing (DLP) technology to fabricate four types of zirconia-based TPMS structures: P-cell, S14, IWP, and Gyroid. For cell proliferation, the four porous TPMS structures outperformed the solid zirconia group (P-cell > S14 > Gyroid > IWP > ZrO2). In vitro assessments on the biological responses and osteogenic properties of the distinct porous surfaces identified the IWP and Gyroid structures as promising candidates for future clinical applications of porous zirconia implants because of their superior osteogenic capabilities (IWP > Gyroid > S14 > P-cell > ZrO2) and mechanical properties (ZrO2 > IWP > Gyroid > S14 > P-cell). Furthermore, the physical properties of the IWP/Gyroid surface had more substantial effects on bone immune regulation by reducing macrophage M1 phenotype polarization while increasing M2 phenotype polarization compared with the solid zirconia surface. Additionally, the IWP and Gyroid groups exhibited enhanced immune osteogenesis and angiogenesis abilities. Collectively, these findings highlight the substantial impact of topology on bone/angiogenesis and immune regulation in promoting bone integration.

Effects of Isochlorogenic Acid on Ewes Rumen Fermentation, Microbial Diversity and Ewes Immunity of Different Physiological Stages.

The effects of isochlorogenic acid (ICGA) on ewes rumen environment, microbial diversity, and immunity at different physiological stages (estrus, pregnancy and lactation) were studied in this experiment. Twenty healthy female Hu lambs of 1.5 months with similar body weight (17.82 ± 0.98 kg) and body condition were selected and randomly divided into two groups: the control group (CON) and the ICGA group (ICGA). The lambs of CON were fed a basal diet, while the lambs of ICGA were supplemented with 0.1% ICGA based on the basal diet. Lambs rumen fermentation characteristics, microbial diversity and immunity at estrus, pregnancy, and lactation stages were determined and analyzed, respectively. The results showed that the rumen pH in CON increased first and then decreased as lambs grew (p < 0.05). However, it showed the opposite change in ICGA. The content of ammonia nitrogen (NH3-N) showed the highest at estrus stage in both groups, but it was significantly higher in ICGA than that in CON (p < 0.05). The Acetic acid/propionic acid (A/P) ratio at estrus stage and the volatile fatty acids (VFAs) at pregnancy stage in ICGA were significantly higher than those of the CON (p < 0.05). The 16S rDNA sequencing analysis showed that the Shannon, Chao 1 and ACE indexes of the ICGA were significantly higher than those of the CON both at estrus and lactation stages (p < 0.05), while they showed higher at the pregnancy stage in CON (p > 0.05). Principal component analysis (PCA) showed that there were significant differences in rumen microorganism structure between CON and ICGA at all physiological stages (p < 0.01). At the phylum level, compared with the CON, Firmicutes relative abundance of three physiological stages decreased (p > 0.05) while Bacteroidota increased (p > 0.05). The relative abundance of Synergistota at estrus stage and Patescibacteria at the lactation stage increased significantly too (p < 0.05). At the genus level, compared with the CON, the relative abundance of Prevotella at three stages showed the highest (p > 0.05), while the relative abundance of Succiniclasticum, unclassified_Selenomonadaceae and Rikenellaceae_RC9_gut_group showed different abundances at different physiological stages in ICGA. Compared with the CON, the lambs of the ICGA showed higher blood IgG, IgM, and TNF- α contents at three physiological stages and higher IL-6 contents at pregnancy stage (p < 0.05). Conclusion: Adding ICGA could regulate ewes rumen fermentation mode at different physiological stages by increasing rumen NH3-N at estrus, VFAs at pregnancy, and the ratio of A/P at lactation. It optimizes rumen microbial flora of different physiological stages by increasing Bacteroidota relative abundance while reducing Firmicutes relative abundance, maintaining rumen microbial homeostasis at pregnant stage, increasing the number of beneficial bacteria in later lactating and ewes blood immunoglobulins content at three physiological stages.

Identification and validation of LINC01322 as a potential prognostic biomarker and oncogene promoting tumor progression in lung adenocarcinoma.

Our study identified a novel long noncoding RNA, LINC01322, that acts as an oncogene in lung adenocarcinoma progression. Cytoplasmic and nuclear RNA purification assays indicated that LINC01322 was localized in the cytoplasm and nucleus. Gene set enrichment analysis revealed the involvement of LINC01322 in the regulation of cell proliferation, migration, and the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. LINC01322 may promote lung adenocarcinoma proliferation and migration through the Janus kinase/signal transducer and activator of transcription signaling pathway. In vitro experiments demonstrated that the knockdown of LINC01322 significantly suppressed lung adenocarcinoma cell proliferation, migration, and activation of the Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway, whereas overexpression had the opposite effects. Inhibition of the Janus kinase 2/signal transducer and activator of transcription 3 pathway activity partially reversed the enhancement of cell proliferation and migration caused by LINC01322 overexpression. In vivo experiments further verified the oncogene role of LINC01322. Altogether, our findings suggest that LINC01322 promotes lung adenocarcinoma progression by activating the Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway and that it could be a therapeutic target.

Antibacterial effects and microarray-based molecular mechanisms of trans-cinnamaldehyde against Porphyromonas gingivalis.

Porphyromonas gingivalis (P. gingivalis) is one of the keystone pathogenic bacteria of periodontitis and peri-implantitis. This study aimed to investigate the antibacterial effects and molecular mechanisms of trans-cinnamaldehyde (TC), a safe extract from natural plants, on P. gingivalis. Minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of TC were determined, and scanning and transmission electron microscopies were used to assess the morphological changes. The overall biomass was estimated, and the metabolic activity of biofilms was determined at different TC concentrations. A microarray-based bioinformatics analysis was performed to elucidate the underlying molecular mechanisms of TC-inhibited P. gingivalis, and significant differences among groups were determined. TC showed an inhibitory effect on the proliferation and survival of planktonic P. gingivalis, of which the MIC and MBC were 39.07 µg/mL and 78.13 µg/mL, respectively. TC also significantly suppressed the formation and metabolic activity of P. gingivalis biofilm. The results of the significant pathways and gene ontology (GO) analyses revealed that TC treatment inhibited two metabolic pathways, accompanied by the downregulation of relative genes of nitrogen metabolism (NrfA, NrfH, and PG_2213) and starch and sucrose metabolism (PG_1681, PG_1682, and PG_1683). Thus, this study confirmed TC to be a natural antimicrobial agent against P. gingivalis and further demonstrated that TC suppressed the microbial activity on P. gingivalis through the disruption of physiological metabolism, which might inhibit the growth and the biofilm formation of P. gingivalis.

Perioperative immune checkpoint inhibition for colorectal cancer: recent advances and future directions.

For colorectal cancer (CRC), surgical resection remains essential for achieving good prognoses. Unfortunately, numerous patients with locally advanced CRC and metastatic CRC failed to meet surgical indications or achieve pathological complete response after surgery. Perioperative therapy has been proven to effectively lower tumor staging and reduce recurrence and metastasis. Immune checkpoint inhibitors (ICIs) have shown unprecedented prolongation of survival time and satisfactory safety in patients with high microsatellite instability/deficient mismatch repair (MSI-H/dMMR), while the therapeutic effect obtained by patients with mismatch repair-proficient or microsatellite stable (pMMR/MSS) was considered minimal. However, recent studies found that certain CRC patients with dMMR/MSI-H presented intrinsic or acquired immune resistance, and pMMR/MSS CRC patients can also achieve better efficacy. Therefore, more predictors are required for screening patients with potential clinical benefits. Since the discovery of synergistic effects between immunotherapy, chemotherapy, and radiotherapy, different immunotherapy-based therapies have been applied to the perioperative therapy of CRC in an increasing number of research. This review comprehensively summarized the past and current progress of different combinations of immunotherapy in perioperative clinical trials for CRC, focusing on the efficacy and safety, and points out the direction for future development.

Comparison of the Retention and Fit of Polyether Ether Ketone Clasps during Fatigue Circulation Tests.

Purpose: The purpose of this study was to evaluate the fit and retention of clasps made of polyether ether ketone (PEEK) or cobalt-chromium alloy (Co-Cr) at different tooth positions in experimental simulations of in vitro wear and removal for 5 years. Methods: Standard crowns of the right mandibular first premolar (44) and first molar (46) were selected, and a circular three-arm clasp was designed, scanned and fabricated. Ten PEEK clasps were used as the experimental group, and 10 Co-Cr clasps were used as the control group. The seating channel was parallel to the side of the abutment base in both groups. The oral environment was simulated, and each clasp was tested in artificial saliva for 7200 cycles while the change in clasp retention force was recorded. The fit before and after the fatigue cycles was measured by the silicone rubber film copying method. Data were statistically analyzed using the independent samples t-test (α = 0.05). Results: Before circulation, the retention forces of the clasps at position 44 were 4.61 ± 0.91 N (PEEK) and 47.50 ± 10.59 N (Co-Cr), and the forces at position 46 were 3.38 ± 0.49 N (PEEK) and 28.79 ± 10.99 N (Co-Cr). After circulation, the retention forces of the clasps at position 44 were 4.15 ± 0.91 N (PEEK) and 13.90 ± 6.59 N (Co-Cr), and the forces at position 46 were 2.93 ± 0.25 N (PEEK) and 11.56 ± 3.93 N (Co-Cr). Before circulation, the fit of each clasp at the reference points (clasp tip, clasp arm, and occlusal rest) was between 41.70 µm and 170.29 µm, and after circulation, they were between 64.05 µm and 182.59 µm. The retention force and fit of the PEEK clasps did not undergo statistically significant changes from before to after circulation (P > 0.05). However, there were statistically significant (P < 0.05) decreases in the retention force of the Co-Cr clasps and the fit of the clasp tip during circulation. In addition, there was a sudden and large change in the retention force of the Co-Cr clasps after approximately 360 cycles. Conclusions: The retention force and suitability of the PEEK clasps met the requirements for clinical use during testing that simulated the in vitro wear and removal procedure for 5 years. Compared with the Co-Cr clasp, the PEEK clasp underwent less fatigue deformation, which makes it feasible for clinical applications.

Analysis of postoperative ocular surface changes and intervention effect after pars plana vitrectomy in meibomian gland dysfunction dry eye patients.

AIM: To observe ocular surface changes after phacovitrectomy in patients with mild to moderate meibomian gland dysfunction (MGD)-type dry eye and track clinical treatment response using a Keratograph 5M and a LipiView interferometer. METHODS: Forty cases were randomized into control group A and treatment group B; the latter received meibomian gland treatment 3d before phacovitrectomy and sodium hyaluronate before and after surgery. The average non-invasive tear film break-up time (NITBUTav), first non-invasive tear film break-up time (NITBUTf), non-invasive measured tear meniscus height (NTMH), meibomian gland loss (MGL), lipid layer thickness (LLT) and partial blink rate (PBR) were measured preoperatively and 1wk, 1 and 3mo postoperatively. RESULTS: The NITBUTav values of group A at 1wk (4.38±0.47), 1mo (6.76±0.70), and 3mo (7.25±0.68) were significantly lower than those of group B (7.45±0.78, 10.46±0.97, and 11.31±0.89; P=0.002, 0.004, and 0.001, respectively). The NTMH values of group B at 1wk (0.20±0.01) and 1mo (0.22±0.01) were markedly higher than those of group A (0.15±0.01 and 0.15±0.01; P=0.008 and P<0.001, respectively); however, there was no difference at 3mo. The LLT of group B at 3mo [91.5 (76.25-100.00)] significantly exceeded that of group A [65.00 (54.50-91.25), P=0.017]. No obvious intergroup difference was found in MGL or PBR (P>0.05). CONCLUSION: Mild to moderate MGD dry eye worsens in the short term after phacovitrectomy. Preoperative cleaning, hot compresses, and meibomian gland massage as well as preoperative and postoperative sodium hyaluronate promote the rapid recovery of tear film stability.

Associations between maternal complications during pregnancy and childhood asthma: a retrospective cohort study.

Background: Studies on the associations between maternal complications during pregnancy and childhood asthma are exclusively conducted in Western countries. The findings are mixed and may not be translated to other populations. We aimed to investigate the associations among the Chinese population and to determine whether the associations were mediated through pre-term birth, caesarean delivery, low birthweight and not breastfeeding in the first 6 months. Methods: We conducted a retrospective cohort study of 166 772 children in Guangzhou, China. Information on maternal gestational hypertension, gestational diabetes and gestational anaemia during pregnancy was extracted from medical records. Ever-diagnosis of asthma in children aged 6-12 years was obtained by questionnaire. Logistic regression models and mediation analyses were used to estimate the adjusted odds ratios (aORs) and 95% confidence intervals for childhood asthma. Results: Gestational hypertension, gestational diabetes and gestational anaemia during pregnancy were associated with an increased risk of ever-diagnosed childhood asthma: aOR 1.48 (95% CI 1.37-1.60), 1.71 (95% CI 1.65-1.78) and 1.34 (95% CI 1.26-1.45), respectively. A stronger association was observed for two or three gestational complications (aOR 2.02 (95% CI 1.93-2.16)) than one gestational complication (aOR 1.64 (95% CI 1.52-1.77)). The aOR for the three gestational complications was 1.35 (95% CI 1.26-1.45), 1.63 (95% CI 1.58-1.70) and 1.32 (95% CI 1.24-1.43), respectively, after controlling for the mediators, including pre-term birth, caesarean delivery, low birthweight and not breastfeeding in the first 6 months. Conclusions: Gestational hypertension, gestational diabetes and gestational anaemia were associated with childhood asthma, and the associations were partially explained by the mediation effects.

In vitro fracture and fatigue resistance of monolithic zirconia crowns fabricated by stereolithography.

STATEMENT OF PROBLEM: Stereolithography has been used to print zirconia ceramic crowns with acceptable dimensional accuracy and fracture force. However, studies that compared the fatigue resistance of zirconia crowns fabricated by stereolithography are lacking. PURPOSE: The purpose of this in vitro study was to compare the fracture and fatigue resistance of monolithic zirconia crowns printed by stereolithography apparatus (SLA) and digital light processing (DLP) with those of zirconia crowns milled by computer numerical control (CNC). MATERIAL AND METHODS: A total of 120 crowns were fabricated (n=40/material) and underwent 0, 104, 105, or 106 dynamic loading cycles of 30 to 300 N in artificial saliva, followed by a static fracture loading test (n=10). After fracture, 1 crown from each group was selected for fractography analysis by scanning electron microscopy (SEM). Data were statistically analyzed through 2-way ANOVA and post hoc analysis for multiple comparisons (α=.05). RESULTS: The 2-way ANOVA results showed that the mean ±standard deviation force at fracture was the highest for CNC (before fatigue loading: 5154 ±568 N, 104: 5735 ±1231 N, 105: 5523 ±797 N, and 106: 6007 ±1258 N), followed by DLP (before fatigue loading: 3381 ±612 N, 104: 4046 ±1146 N, 105: 2929 ±559 N, and 106: 3223 ±739 N), and the lowest for SLA (before fatigue loading: 2956 ±598 N, 104: 2757 ±421 N, 105: 3326 ±391 N, and 106: 3103 ±246 N) (P<.01). The fracture force of the crowns was not significantly affected by the number of fatigue cycles (P>.05). Fractography analysis showed that the number of arrest lines increased for crowns of all 3 materials. SEM images also showed the steps of SLA and DLP from their layer-by-layer printing and small cracks between layers of SLA after 106 loading cycles. CONCLUSIONS: The fracture force of monolithic zirconia crowns milled by CNC was significantly higher than that of zirconia crowns printed by stereolithography. Zirconia crowns printed by SLA and DLP could withstand typical clinical conditions, and their fracture and fatigue resistance exceeded the clinically estimated average occlusal forces.

Comprehensive Analysis of the Longissimus Dorsi Transcriptome and Metabolome Reveals the Regulatory Mechanism of Different Varieties of Meat Quality.

The beef quality significantly varies between breeds. Pingliang Red Cattle resembles Wagyu in fat deposition and flavor. To screen key factors affecting beef quality, we performed meat quality trait testing, RNA-seq, and metabolomics on the longissimus dorsi of Pingliang Red Cattle, Wagyu cross F1 generation, and Simmental cattle. The gene and metabolite expression profiles were similar between Pingliang Red Cattle and Wagyu cross F1 generation. Genes such as FASN, ACACA, PLIN1, and FABP4 were significantly upregulated in the Pingliang Red Cattle and Wagyu cross F1 generation (P < 0.05). Similarly, numerous metabolites, such as 3-iodo-l-tyrosine, arachidonic acid, and cis-aconitate, which may improve the beef quality such as fat deposition and tenderness, were found in higher levels in the Pingliang Red Cattle and Wagyu cross F1 generation. This study revealed differences in the transcriptional and metabolic levels between Pingliang Red Cattle and premium beef breeds, suggesting that Pingliang Red Cattle harbors the genetic potential for breeding high-grade beef cattle.

Effects of different proportions of stevia stalk on nutrient utilization and rumen fermentation in ruminal fluid derived from sheep.

Background: Stevia straw is a byproduct of sugar crop stevia. It is a good feed material because of richness in nutrients and active substances (steviosides and flavonoids). However, due to improper utilization such as piling, burning and so on, it became a large amount of wasted straw resources and lead to environmental pollution. Methods: We added 0%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.5% of stevia stalk to study the effects of different stevia stalk concentrations on nutrient utilization and rumen fermentation in sheep (based on sheep diet). In vitro fermentation method was used, with 17 repetitions for each treatment. All fermentation substrate based on sheep diet with different stevia stalk concentrations were fermented for 2 h, 6 h, 12 h, 24 h and 48 h, then the gas production, dry matter degradability (DMD), crude protein degradability (CPD), neutral detergent fiber degradability (NDFD), acid detergent fiber degradability (ADFD), pH, ammonia nitrogen (NH3-N) and volatile fatty acids (VFAs) were determined. Results: The results showed that at different fermentation time, the change trend of gas production in each teatment was basically same, but the maximum occurred in 1.0% treatment at 48 h. The DMD, CPD, NDFD and ADFD of sheep diets increased with fermentation time increasing, especially the CPD48h, NDFD48h and ADFD48h of diets in 0.8%, 1.0% and 1.5% treatments were significantly higher than those in control (P < 0.05). The pH of fermentation substrate in each treatment remained within the normal range of 6.21∼7.25. NH3-N24h-48hin 0.8%, 1.0% and 1.5% treatments were higher than that in control. At 6 h-12 h, the total acid content of 0.8% and 1.0% treatments were significantly higher than those of other treatments (P < 0.05), it reached the highest in 1.0% treatment. According to overall evaluation, effect ranking of stevia stalk on sheep nutrient utilization was as follows: 1.0% >0.8% >1.5% >0.4% >0.6% >0.2%. Overall, 1.0% stevia stalk could promote nutrient degradation and sheep rumen fermentation.

[Accuracy evaluation of recording maxillomandibular relation with the mandibular kinesiography].

PURPOSE: To evaluate the accuracy of recording maxillomandibular position relationship with the mandibular kinesiography by surface electromyography analysis of the masticatory muscles and MRI of temporomandibular joint (TMJ). METHODS: Eight edentulous patients were selected randomly, and the maxillomandibular position relationship was determined by mandibular kinesiography and traditional method respectively. Then the corresponding bite plates were made to wear. Surface electromyography test of left and right anterior temporal and masseter muscle and magnetic resonance scan of the temporomandibular joint were performed on closed mouth position to record peak potentials, calculate asymmetric indexes, analyze the position relationship between the joint disc and the condyle, measure TMJ joint space. SPSS 18.0 software package was used for data analysis. RESULTS: There was no significant difference in the amplitude of the anterior temporal or masseter muscle, the asymmetry index of total, asymmetry index of masseter, asymmetry index of temporalis, the anterior, upper and posterior spaces of TMJ between the mandibular kinesiography group and the traditional group (P＞0.05). The temporomandibular articular disc process relationship of the two groups were conformed to the normal physiological position relationship. CONCLUSIONS: Mandibular kinesiography could accurately record the maxillomandibular position relationship in patients with edentulous jaws.

Effects of Dietary Quinoa Seeds on Cecal Microorganisms and Muscle Fatty Acids of Female Luhua Chickens.

To study the effects of adding quinoa seed (raw grain) to the diet of the Luhua chicken on the cecal microorganism abundance and fatty acid composition of muscle, 120 49-day-old healthy female dewormed Luhua chickens (body weight 1476.21 ± 101.39 g) were randomly divided into 4 groups, with 3 replicates in each group and 10 chickens in each repetition. The control group (CK group) was fed a basal diet and the experimental groups were fed with 4% (Q4), 8% (Q8), and 12% (Q12) quinoa seed (raw grain) added to the basal diet for 75 days. After 121 days of age, the animals were slaughtered and the 16S rRNA characteristics of cecal flora, as well as composition and content of fatty acids in muscle, were determined and analyzed. The content of unsaturated fatty acids (UFAs), docosahexaenoic acid (C22:6n3; DHA) and n-3 polyunsaturated fatty acids (n-3 PUFAs) in the breast and leg muscles significantly increased in the experimental groups supplemented with quinoa seeds (p < 0.05). However, the content of saturated fatty acids (SAFs) and ratio of n-6/n-3 in breast muscle and leg muscle significantly decreased (p < 0.05). In addition, adding a certain percentage of quinoa seeds in the diet can also affect the community composition and content of microorganisms in the ceca of Luhua chickens. At the phylum level, the Proteobacteria, Actinobacteria, Synergistetes and Melainabacteria in experimental groups (Q4, Q8 and Q12) were significantly lower than those in the CK group (p < 0.05). At the genus level, Desulfovibrio, Synergistes, Olsenella, Parabacteroides, Mailhella, Sutterella and Ruminiclostridiu in group Q4 were significantly lower than those in group CK (p < 0.05) while Faecalibacterium in Q8 group, and Lawsonia and Faecalibacterium in Q12 group were significantly higher than those in the CK group (p < 0.05). Enrichment analysis of the microbial function showed that compared with the CK group, Metabolism and Enzyme Families were significantly enriched in the Q4 group (p < 0.05). Cellular Processes and Signaling were significantly enriched in the Q8 group (p < 0.05). The association analysis of fatty acids with microorganisms showed that the abundance of Faecalibacterium, Lawsonia and Meagmonas was significantly correlated with partial SFAs and UFAs (p < 0.05). In conclusion, adding quinoa seeds to diets significantly increased the content of muscle DHA, UFAs and n-3 PUFAs. The content of SAFs and the n-6/n-3 ratio were significantly reduced. Taken together, quinoa can effectively improve the cecal microbiota structure, inhibit the number of harmful bacteria and increase the number of beneficial bacteria, regulating the intestinal environment and promoting the body health of female Luhua chickens.

Negative effects of enlarging internal limiting membrane peeling for idiopathic macular hole surgery.

AIM: To observe the effects of the different extents of internal limiting membrane (ILM) peeling on the surgical success and anatomical and functional outcomes of idiopathic macular hole (IMH). METHODS: In this retrospective cohort study, 36 patients were reviewed and divided into two groups according to the extent of ILM peeling: group A (18 patients), with the peeling area within one-half of the optic disc macular distance as the radius; group B (18 patients), with the peeling area larger than that of group A but did not exceed the optic disc macular distance as the radius. The main outcomes included the best corrected visual acuity (BCVA), light-adaptive electroretinography, macular hole (MH) closure rate, central macular thickness (CMT), retinal nerve fiber layer (RNFL) and ganglion cell complex (GCC) thickness [nine regions based on the Early Treatment of Diabetic Retinopathy Study (ETDRS) ring] before and 1, 3, and 6mo after surgery. RESULTS: The closure rate was 94.4% (17/18) both in groups A and B. The BCVA in both groups improved significantly compared with the preoperative values, but there was no difference between the two groups. The b-wave amplitude of the electroretinogram analysis was significantly improved in both groups compared to that of the preoperative period, with a greater increase in group A than in group B at 6mo (P=0.017). The CMT in both groups gradually decreased after surgery, and there was no difference between the two groups. The RNFL thickness of the temporal outer ring region in group B was significantly lower than that in group A at 3 and 6mo after surgery (P=0.010, 0.032). The GCC thickness of the temporal outer ring region in group B was significantly lower than that in group A at 6mo after surgery (P=0.038). CONCLUSION: Enlarging the extent of ILM peeling doesn't affect the IMH closure rate and visual acuity recovery, but the greater the extent of peeling, the greater the damage to the inner retinal structures.

Effects of Dietary Oregano Essential Oil on Cecal Microorganisms and Muscle Fatty Acids of Luhua Chickens.

This experiment was conducted to investigate the effects of oregano essential oil on the cecal microorganisms and muscle fatty acids of Luhua chickens. One hundred and twenty 49-day-old healthy dewormed Luhua chickens were randomly divided into four groups with three replicates per group and ten chickens per replicate. The corn−quinoa and soybean meal diets were supplemented with 0 (Q8 group), 50 (QO50 group), 100 (QO100 group) and 150 mg·kg−1 (QO150 group) of oregano essential oil, respectively, and the experiment lasted for 75 days. The composition of intestinal flora was detected by Illumina sequencing of the 16S rRNA V4 region, and the composition and content of fatty acids in the muscles were analyzed by gas chromatography. The results showed that dietary oregano essential oil can effectively increase the contents of elaidic acid (C18:ln9t), polyunsaturated fatty acids (PUFAs) and n-3 polyunsaturated fatty acids (n-3 PUFAs) in breast muscle tissues. However, the fatty acid composition and PUFA content in leg muscle tissues were not significantly improved. According to a 16S rRNA high-throughput sequencing analysis, dietary oregano essential oil supplementation with a certain concentration can change the cecal microbial community composition of broilers. At the phylum level, Elusimicrobia in the QO150 group was significantly lower than that in Q8 group (p < 0.05). At the genus level, Phascolarctobacterium, Parasutterella and Bilophila in the experimental groups (QO50, QO100 and QO150) were significantly lower than those in the Q8 group (p < 0.05). An enrichment analysis of the microbial function found that the amino acid metabolism, energy metabolism, metabolism, signal transduction and genetic information processing were mainly enriched in the experimental groups, which promoted the digestion and absorption of nutrients and enhanced intestinal barrier functioning. An analysis of the association between fatty acids and microbes found that the abundance of microbiota was significantly correlated with partially saturated fatty acids (SFAs) and unsaturated fatty acids (UFAs) (p < 0.05). In conclusion, the dietary addition of oregano essential oil can effectively improve cecal microbial community composition, promote the digestion and absorption of nutrients, and enhance intestinal barrier functioning. It can significantly improve the content of some fatty acids, and there was a certain correlation between caecum microorganisms and fatty acid deposition in muscles.

Characterization of circRNA-miRNA-mRNA networks regulating oxygen utilization in type II alveolar epithelial cells of Tibetan pigs.

Understanding the signaling pathway regulatory mechanisms in type II alveolar epithelial (ATII) cells, the progenitor cells responsible for proliferating and regenerating type I alveolar epithelial (ATI) and ATII cells, in Tibetan pigs is beneficial for exploring methods of preventing and repairing cellular damage during hypoxia. We simulated a hypoxic environment (2% O2) for culture ATII cells of Tibetan pigs and Landrace pigs, with cells cultured under normoxic conditions (21% O2) as a control group, and performed integrated analysis of circular RNA (circRNA)-microRNA (miRNA)-messenger RNA (mRNA) regulatory axes by whole-transcriptome sequencing. Functional enrichment analysis indicated that the source genes of the differential expressed circRNAs (DEcircRNAs) were primarily involved in cell proliferation, cellular processes, and cell killing. A series of DEcircRNAs were derived from inhibitors of apoptosis proteins and led to a key autonomous effect as modulators of cell repair in Tibetan pigs under hypoxia. The significant higher expression of COL5A1 in TL groups may inhibited apoptosis of ATII cells in Tibetan pigs under lower oxygen concentration, and may lead their better survive in the hypoxia environment. In addition, a competing endogenous RNA (ceRNA) network of functional interactions was constructed that included novel_circ_000898-ssc-miR-199a-5p-CAV1 and novel_circ_000898-ssc-miR-378-BMP2, based on the node genes ssc-miR-199a-5p and ssc-miR-378, which may regulate multiple miRNAs and mRNAs that mediate endoplasmic reticulum (ER) stress-induced apoptosis and inflammation and attenuate hypoxia-induced injury in ATII cells under hypoxic conditions. These results broaden our knowledge of circRNAs, miRNAs, and mRNAs associated with hypoxia and provide new insights into the hypoxic response of ATII cells in Tibetan pigs.

Effects of different fertilizers on nutrient quality and mineral elements in different economic forage groups in Qilian Mountain alpine meadows.

The objective of this study was to examine the responses of nutrient quality and mineral elements of forages in different economic groups forages in the Qilian Mountains alpine meadows to different fertilization treatments. Eight fertilization treatments, potassium (K), nitrogen (N), zinc (Z), boric (B), copper (Cu), phosphorus (P), molybdenum (Mo), and selenium (Se), were sprayed onto forage roots, and no fertilizer was applied as a blank control (CK), with four replicates in each group. The forage trace element contents and nutritional quality of each economic group in to different treatments were determined in mid-August, and the trace element surplus and deficiency were analyzed. Compared with that in the control, the forage crude protein (CP) content in different economic groups under different fertilization treatments increased and fluctuated within the range of 7.7%-23.94%. The dry matter digestibility (DMD) ranged from 38.78% to 77.34% and showed no significant differences in DMD among treatments (P > 0.05). The DMD of legume forages was significantly higher than those of Gramineae grasses and sedges; there were no significant differences in forage acid detergent fiber (ADF) between any treatments and the control (P > 0.05), but ADF showed a decreasing trend in the treatments compared with that in the control. The P content increased by 81.81% in legumes (P < 0.05); however, it decreased by 38.46% and 61.29% in wilted grass and forbs (P < 0.05) and increased in Gramineae grasses and legumes under N treatment by 92.86% and 50% (P < 0.05), respectively. The Cu content under Cu, N, Zn, B and Se treatments increased significantly by 33.81%∼346.49% compared with that in the control. There were no significant differences in Zn content among the economic groups under different treatments (P > 0.05), but legumes showed better absorption of Zn. Consequently, the forages in this study were evaluated as having medium Cu (8.1-20 mg/kg), medium Se (0.11-1.0 mg/kg), and medium (30.1-60 mg/kg) to high (60.1-100 mg/kg) Zn contents. Comprehensive analysis showed that the application of different fertilizers could increase the CP and DMD of various forages, reduce ADF, and effectively improve the nutritional quality of pastures. The contents of N, Cu, Zn, and Se in forages increased under all treatments, and the contents of all forage elements were at appropriate levels. Applying K, N, P, Cu and Mo is beneficial to dominant species in Qilian Mountains alpine meadows.