Effects of kinds of additives on fermentation quality, nutrient content, aerobic stability, and microbial community of the mixed silage of king grass and rice straw.

To investigate the effects of kinds of additives on silage quality, the mixture of king grass and rice straw was ensiled with addition of sucrose, citric acid and malic acid at the levels of 0, 1 and 2%, being blank control (CK), citric acid groups (CA1, CA2), malic acid groups (MA1, MA2), citric acid + malic acid groups (CM1, CM2), sucrose groups (SU1, SU2), mainly focusing on fermentation quality, nutrient content, aerobic stability and microbial community of the silages. The results showed that the addition of sucrose decreased (p < 0.05) pH and increased the content of water soluble carbohydrate (p < 0.05). The sucrose groups and mixed acid groups also had a lower (p < 0.01) neutral detergent fiber content. The addition of citric acid and the mixed acid increased (p < 0.01) the aerobic stability of the silage, reduced the abundance of Acinetobacter, and the addition of citric acid also increased the abundance of Lactiplantibacillus. It is inferred that citric acid and malic acid could influence fermentation quality by inhibiting harmful bacteria and improve aerobic stability, while sucrose influenced fermentation quality by by promoting the generation of lactic acid. It is suggested that the application of citric acid, malic acid and sucrose would achieve an improvement effect on fermentation quality of the mixed silage.

Identification and effective regulation of scarb1 gene involved in pigmentation change in autotetraploid Carassius auratus.

The autotetraploid Carassius auratus (4nRR, 4 n=200, RRRR) is derived from whole-genome duplication of Carassius auratus red var. (RCC, 2 n=100, RR). In the current study, we demonstrated that chromatophores and pigment changes directly caused the coloration and variation of 4nRR skin (red in RCC, brownish-yellow in 4nRR). To further explore the molecular mechanisms underlying coloration formation and variation in 4nRR, we performed transcriptome profiling and molecular functional verification in RCC and 4nRR. Results revealed that scarb1, associated with carotenoid metabolism, underwent significant down-regulation in 4nRR. Efficient editing of this candidate pigment gene provided clear evidence of its significant role in RCC coloration. Subsequently, we identified four divergent scarb1 homeologs in 4nRR: two original scarb1 homeologs from RCC and two duplicated ones. Notably, three of these homeologs possessed two highly conserved alleles, exhibiting biased and allele-specific expression in the skin. Remarkably, after precise editing of both the original and duplicated scarb1 homeologs and/or alleles, 4nRR individuals, whether singly or multiply mutated, displayed a transition from brownish-yellow skin to a cyan-gray phenotype. Concurrently, the proportional areas of the cyan-gray regions displayed a gene-dose correlation. These findings illustrate the subfunctionalization of duplicated scarb1, with all scarb1 genes synergistically and equally contributing to the pigmentation of 4nRR. This is the first report concerning the functional differentiation of duplicated homeologs in an autopolyploid fish, substantially enriching our understanding of coloration formation and change within this group of organisms.

Effects of Cellulase and Xylanase on Fermentation Characteristics, Chemical Composition and Bacterial Community of the Mixed Silage of King Grass and Rice Straw.

This study was to investigate the effects of cellulase and xylanase on fermentation characteristics, nutrient composition and the bacterial community of the mixed silage of king grass and rice straw. Lab-scale bag silage was produced and seven groups were studied: blank control (CK); added 1%, 2% cellulase (CE1, CE2); added 1%, 2% xylanase (XY1, XY2); and added 0.5% cellulase +0.5% xylanase, 1% cellulase +1% xylanase (CX1, CX2). The results showed that the application of additives in six treated groups exerted a positive effect on lactic acid (LA) content and their pH values decreased significantly (p < 0.05). The addition of cellulase and xylanase decreased (p < 0.05) the content of neutral detergent fiber (NDF) and acid detergent fiber (ADF) significantly and increased (p < 0.01) the crude protein (CP) and water-soluble carbohydrate (WSC) content. Filter paper enzyme activity (FPA) declined and xylanase activity (XA) intensified (p < 0.05) as ensiling was prolonged, where most of the enzymatic treatments (especially XY2, CX2) resulted in increased enzyme activities. Moreover, the addition of cellulase and xylanase reduced the abundance of harmful bacteria such as Acinetobacter and Klebsiella and increased the abundance of lactic acid bacteria such as Lacticaseseibacillus, Lactiplantibacillus. In conclusion, the addition of cellulase and xylanase would improve fermentation quality and nutrient preservation via altering the bacterial community, with 1% cellulase or complex enzyme best.

Mapping multi-seasonal habitats of giant pandas to identify seasonal shifts.

As a flagship species of biodiversity conservation globally, the giant panda has seasonal migration to cope with seasonal changes in available resources. Here, we have mapped the spatial distribution of multi-seasonal habitats of the giant panda across the Baishuijiang reserve in China. Results show that the spatial patterns are different in different seasons, generally, large patches are observed in the western part, while staggered clusters occur in the middle and eastern parts. That is, suitable habitats for giant pandas are mostly distributed in the west part. More than 75% of the predicted suitable habitats are within the core zone of the reserve year-round, indicating the core zone essentially meet giant panda's ecological needs, although this range could potentially be expanded. This study provides valuable insights into the spatiotemporal migration patterns of endangered species and helps to guide conservation planning.

Effects of Dietary Selenium Yeast Supplementation in Pregnant Cashmere Goats on the Development of Offspring Hair Follicles.

The objective of this study was to investigate the effects of maternal dietary selenium yeast (SY) supplementation during pregnancy on the hair follicle development of kids. Sixty pregnant Hanshan white cashmere goats were randomly divided into the con group (fed with a basal diet) and the SY group (fed with a basal diet with 0.4 mg/kg SY). SY was supplemented during the pregnancy until the birth of the kids. The growth performance, cashmere performance, hair follicle characteristics, and serum antioxidant capacity of the kids were periodically determined. The results showed that the birth weight of the kids in the SY group was significantly higher than that in the con group (p < 0.05), and the average weight at 15 days, 1 month, 3 months, and 5 months of age increased by 13.60%, 8.77%, 8.86%, and 3.90%, respectively (p > 0.05). The cashmere fineness at early birth was dramatically reduced with SY supplementation (p < 0.001), whereas cashmere length and production were significantly increased at 5 months of age (p < 0.05). Histology assays indicated that the primary hair follicles were fully developed at birth, and there was no significant difference in the number of primary hair follicles between the two groups (p > 0.05). The number of secondary hair follicles and the number and density of active secondary hair follicles in the SY group at 15 days were significantly higher than those in the con group (p < 0.05) and were increased by 11.18%, 6.18%, and 22.55% at 5 months of age, respectively (p > 0.05). The serum antioxidant capacity analysis revealed that the SY group had higher levels of T-AOC, SOD, CAT, and GSH-Px activities and lower levels of MDA (p > 0.05). These results reveal that the maternal dietary supplementation of SY in gestation can promote the morphogenesis and maturation of secondary hair follicles and increase the number and density of secondary hair follicles by enhancing the body's antioxidant capacity, contributing to the improvement of cashmere quality and yield.

Improved maximum likelihood method for P-S-N curve fitting method with small number specimens and application in T-welded joint.

In fatigue data analysis, fitting accurate P-S-N curve is problematic if only a small number of specimen is available, especially to evaluate the relationship between the stress level and the standard deviation. This paper proposes a sample information reconstruction method that can effectively solve this problem. Based on this method and the life equivalent principle, a new maximum likelihood method (which is abbreviated to improved maximum likelihood method) is proposed for P-S-N curve fitting. T-joint specimens of Q450NQR1 steel were fabricated and tested, then the P-S-N curves was fitted by the improved maximum likelihood method, least square method, maximum likelihood method, standard BS7608 and standard IIW. Finally, P-S-N curves by three methods and two standards are compared and analyzed. The results show that the relevant parameters of the P-S-N curve with 99.9% survival probability fitted by the improved maximum likelihood method are similar to those in the two standards, and it is indicated that the improved maximum likelihood method is a better way for P-S-N curve fitting with the small number of fatigue test specimens.

Single-Cell RNA-Seq Analysis Identifies Angiotensinogen and Galanin as Unique Molecular Markers of Acinar Cells in Murine Salivary Glands.

The submandibular gland (SMG) and sublingual gland (SLG) are two of three major salivary glands in mammals and comprise serous and mucous acinar cells. The two glands share some functional properties, which are largely dependent on the types of acinar cells. In recent years, while ScRNA-seq (single-cell sequencing) with a 10 × platform has been used to explore molecular markers in salivary glands, few studies have examined the acinar heterogeneity and unique molecular markers between SMG and SLG. This study aimed to identify the molecular markers of acinar cells in the SLG and SMG. We performed ScRNA-seq analyses in 4-week-old mice and verified the screened molecular markers using reverse transcription-quantitative real-time PCR, immunohistochemistry, and immunofluorescence. Our results showed prominently heterogeneous acinar cells, although there was great similarity in the cluster composition between the two glands at 4 weeks. Furthermore, we demonstrated that Agt is a specific marker of SMG serous acinar cells, whereas Gal is a specific marker of SLG mucous acinar cells. Trajectory inference revealed that Agt and Gal represent two types of differential acinar cell clusters during late development in adults. Thus, we reveal previously unknown specific markers for salivary acinar cell diversity, which has extensive implications for their further functional research.

Successful regression of newly formed corneal neovascularization by subconjunctival injection of bevacizumab in patients with chemical burns.

Purpose: To investigate the effect and timing of subconjunctival bevacizumab injection on inhibiting corneal neovascularization (CorNV) in patients after chemical burns. Methods: Patients with CorNV secondary to chemical burns were involved. Two subconjunctival injections of bevacizumab (2.5 mg/0.1 mL per involved quadrant) with an interval of 4 weeks were administered, and followed up a year. The area occupied by neovascular vessels (NA), accumulative neovascular length (NL), mean neovascular diameter (ND), best-corrected visual acuity (BCVA) and intraocular pressure (IOP) were evaluated. Complication was also recorded. Results: Eleven patients with CorNV were involved. Eight patients had a history of surgery (four had amniotic grafts, one had keratoplasty, and three had amniotic grafts and keratoplasty). Decreasing in NA, NL, and ND were statistically significant at each time point compared to the baseline (p < 0.01). CorNV that developed within 1 month was considerably regressed, and vessels with fibrovascular membranes were found to be narrower and shorter than pretreatment. BCVA improved in five patients (from one to five lines), remained unchanged in five patients, and decreased in one patient compared to pretreatment. Conclusion: Subconjunctival bevacizumab injection has a particular potential for the regression of CorNV, especially newly formed within 1 month in patients after chemical burns.

CD93 Ameliorates Diabetic Wounds by Promoting Angiogenesis via the p38MAPK/MK2/HSP27 Axis.

OBJECTIVE: Diabetic wounds are a complication of diabetes mellitus, which is characterised by microcirculation dysfunction caused by decreased local blood supply and insufficient metabolic exchange. Clinically, in addition to glycaemic control, the most important treatment for diabetic wounds is to promote local angiogenesis, which accelerates wound healing. The authors previous study demonstrated that CD93, which is specifically expressed on vascular endothelial cells (ECs), redundantly regulates angiogenesis in zebrafish, suggesting that CD93 is a potential angiogenic molecule. However, the role of CD93 in diabetic wounds has not yet been elucidated. METHODS: The angiogenic effects of CD93 were studied from four aspects: exogenous, endogenous, in vitro, and in vivo. CD93 recombinant protein was used in microvascular ECs and in mice to observe angiogenesis in vitro and in vivo. The wound model was established in CD93-/- and wild type diabetic mice, and the degree of wound healing as well as the amount and maturity of neovascularisation were investigated. The possible mechanism of CD93 in angiogenesis was determined by CD93 overexpression in cultured ECs. RESULTS: CD93 recombinant protein was found to exogenously promote tube formation and sprouting of ECs. It also recruited cells to promote the formation of vascular like structures in subcutaneous tissue and accelerated wound healing by optimising angiogenesis and re-epithelisation. Furthermore, CD93 deficiency was observed to delay wound repair, characterised by reduced neovascularisation, vascular maturity, and re-epithelisation level. Mechanically, CD93 activated the p38MAPK/MK2/HSP27 signalling pathway, positively affecting the angiogenic functions of ECs. CONCLUSION: This study demonstrated that CD93 promotes angiogenesis both in vitro and in vivo and that its angiogenic role in vitro is mediated by the p38MAPK/MK2/HSP27 signalling pathway. It was also found that CD93 exerts beneficial effects on wound healing in diabetic mice by promoting angiogenesis and re-epithelisation.

Exopolysaccharides metabolism and cariogenesis of Streptococcus mutans biofilm regulated by antisense vicK RNA.

Background: Streptococcus mutans (S. mutans) is a pivotal cariogenic pathogen contributing to its multiple virulence factors, one of which is synthesizing exopolysaccharides (EPS). VicK, a sensor histidine kinase, plays a major role in regulating genes associated with EPS synthesis and adhesion. Here we first identified an antisense vicK RNA (ASvicK) bound with vicK into double-stranded RNA (dsRNA). Objective: This study aims to investigate the effect and mechanism of ASvicK in the EPS metabolism and cariogenesis of S. mutans. Methods: The phenotypes of biofilm were detected by scanning electron microscopy (SEM), gas chromatography-mass spectrometery (GC-MS) , gel permeation chromatography (GPC) , transcriptome analysis and Western blot. Co-immunoprecipitation (Co-ip) assay and enzyme activity experiment were adopted to investigate the mechanism of ASvicK regulation. Caries animal models were developed to study the relationship between ASvicK and cariogenicity of S. mutans. Results: Overexpression of ASvicK can inhibit the growth of biofilm, reduce the production of EPS and alter genes and protein related to EPS metabolism. ASvicK can adsorb RNase III to regulate vicK and affect the cariogenicity of S. mutans. Conclusions: ASvicK regulates vicK at the transcriptional and post-transcriptional levels, effectively inhibits EPS synthesis and biofilm formation and reduces its cariogenicity in vivo.

Melatonin promotes the development of the secondary hair follicles by regulating circMPP5.

BACKGROUND: The quality and yield of cashmere fibre are closely related to the differentiation and development of secondary hair follicles in the skin of cashmere goats. The higher the density of secondary hair follicles, the higher the quality and yield of cashmere from the fleece. Development of secondary hair follicles commences in the embryonic stage of life and is completed 6 months after birth. Preliminary experimental results from our laboratory showed that melatonin (MT) treatment of goat kids after their birth could increase the density of secondary hair follicles and, thus, improve the subsequent yield and quality of cashmere. These changes in the secondary hair follicles resulted from increases in levels of antioxidant and expression of anti-apoptotic protein, and from a reduction in apoptosis. The present study was conducted to explore the molecular mechanism of MT-induced secondary hair follicle differentiation and development by using whole-genome analysis. RESULTS: MT had no adverse effect on the growth performance of cashmere kids but significantly improved the character of the secondary hair follicles and the quality of cashmere, and this dominant effect continued to the second year. Melatonin promotes the proliferation of secondary hair follicle cells at an early age. The formation of secondary hair follicles in the MT group was earlier than that in the control group in the second year. The genome-wide data results involved KEGG analysis of 1044 DEmRNAs, 91 DElncRNAs, 1054 DEcircRNAs, and 61 DEmiRNAs which revealed that the mitogen-activated protein kinase (MAPK) signaling pathway is involved in the development of secondary hair follicles, with key genes (FGF2, FGF21, FGFR3, MAPK3 (ERK1)) being up-regulated and expressed. We also found that the circMPP5 could sponged miR-211 and regulate the expression of MAPK3. CONCLUSIONS: We conclude that MT achieves its effects by regulating the MAPK pathway through the circMPP5 sponged the miR-211, regulating the expression of MAPK3, to induce the differentiation and proliferation of secondary hair follicle cells. In addition there is up-regulation of expression of the anti-apoptotic protein causing reduced apoptosis of hair follicle cells. Collectively, these events increase the numbers of secondary hair follicles, thus improving the production of cashmere from these goats.

In Situ Degradation Kinetics of 25 Feedstuffs and the Selection of Time Points in Mathematical Statistics.

Rumen degradation kinetics of 25 feedstuffs (six protein feeds, nine energy feeds and ten roughages) were first determined using the nylon bag technique in situ and the differences of degradation characteristics fitted with five or seven time points measuring data were evaluated with the goodness of fit (R2) of degradation curves. Protein and energy feeds were incubated for 2, 4, 8, 16, 24, 36, 48 h, roughages were incubated for 4, 8, 16, 24, 36, 48, 72 h, where three and six data sets of five time points were screened out, respectively. Only the degradation parameters a (rapidly degraded proportion), b (slowly degraded proportion) and c (degradation rate of slowly degraded proportion) of several feeds at five time points were significantly different from those at seven time points (p < 0.05), and the others were not significant (p > 0.05). The R2 of the degradation curves obtained at five time points was closer to 1, indicating that the fitting obtained at five time points was more accurate in predicting the real-time rumen degradation rate of feed. These results indicate that it is feasible to determine the rumen degradation characteristics of feedstuffs by only setting five measuring time points.

Hair Follicle Development and Cashmere Traits in Albas Goat Kids.

The objectives of this trial were to study the growth and development of hair follicles and cashmere traits in cashmere goats and to provide a theoretical basis for the regulation of secondary hair follicle development and the scientific breeding selection of cashmere goats. Twelve single-fetal female kids were selected as research objects. A long-term tracking plan was created to regularly determine their growth performance, cashmere performance, and hair follicle traits. The results showed no significant difference in live weight after the first and second combing. The cashmere yield and unit yield of the first combing were significantly higher than those of the second combing (p < 0.05). Sections of hair follicles showed that the primary hair follicles are almost fully developed by 1 month, and the secondary hair follicles are fully developed by 5-6 months after birth. The primary hair follicle density (PFD) and secondary hair follicle density (SFD) were highest at birth and decreased within 1 month; and SFD was stable at 5-6 months of age. The change of MSFD took a maximum time of 2 to 3 months. The S:P increase reached its peak at 6 months. BMP4 expression increased with time. FGF2, FGF21 and BMP7 were higher at 3 months old than at the other two-time points. In conclusion, this study determined the total development time of primary and secondary hair follicles from morphology and speculated that FGF2, FGF21, and BMP7 may play a regulatory role in developing secondary hair follicles. Therefore, the period from birth to 6 months of age was the best time to regulate secondary hair follicle development in cashmere goats kids. The traits of the hair follicle and cashmere at 6 months of age could be breeding selection indicators for cashmere goats.

Melatonin Promotes the Development of Secondary Hair Follicles in Adult Cashmere Goats by Activating the Keap1-Nrf2 Signaling Pathway and Inhibiting the Inflammatory Transcription Factors NFκB and AP-1.

Exogenous melatonin (MT) has been used to promote the growth of secondary hair follicles and improve cashmere fiber quality, but the specific cellular-level mechanisms involved are unclear. This study was carried out to investigate the effect of MT on the development of secondary hair follicles and on cashmere fiber quality in cashmere goats. The results showed that MT improved secondary follicle numbers and function as well as enhanced cashmere fiber quality and yield. The MT-treated goat groups had high secondary-to-primary ratios (S:P) for hair follicles, greater in the elderly group (p < 0.05). Antioxidant capacities of secondary hair follicles improved fiber quality and yield in comparison with control groups (p < 0.05/0.01). Levels of reactive oxygen and nitrogen species (ROS, RNS) and malondialdehyde (MDA) were lowered (p < 0.05/0.01) by MT. There was significant upregulation of antioxidant genes (for SOD-3; GPX-1; NFE2L2) and the protein of nuclear factor (Nrf2), and downregulation of the Keap1 protein. There were significant differences in the expression of genes for secretory senescence-associated phenotype (SASP) cytokines (IL-1ß, IL-6, MMP-9, MMP-27, CCL-21, CXCL-12, CXCL-14, TIMP-1,2,3) plus their protein of key transcription factors, nuclear factor kappa B (NFκB) and activator protein-1 (AP-1), in comparison with the controls. We concluded that MT could enhance antioxidant capacity and reduce ROS and RNS levels of secondary hair follicles through the Keap1-Nrf2 pathway in adult cashmere goats. Furthermore, MT reduced the expression of the SASP cytokines genes by inhibiting the protein of NFκB and AP-1 in the secondary hair follicles in older cashmere goats, thus delaying skin aging, improving follicle survival, and increasing the number of secondary hair follicles. Collectively, these effects of exogenous MT enhanced the quality and yield of cashmere fibers, especially at 5-7 years old.

Supplementary Feeding of Grazing Inner Mongolian Cashmere Goats during Pregnancy-Based on "Nutrient Requirements of Cashmere Goats".

This study aimed to conduct precise supplementation for pregnant cashmere goats under grazing based on the feeding standard. Eight Inner Mongolian pregnant cashmere goats of near-average body weight were selected at early gestation (44.41 ± 4.03 kg) and late gestation (46.54 ± 4.02 kg) to measure their nutrient intake. Then, two pregnant cashmere goat flocks, No. 10 (control group, on-farm supplement) and No. 11 (supplemented group, supplement based on standard), with the same goat herd structure and grassland type, were chosen to conduct the supplemental feeding experiment. The results showed that pregnant cashmere goats lacked daily the intake of dry matter, digestive energy, crude protein and most essential mineral elements under grazing. After supplemental feeding, the supplementation based on the feeding standard increased the cashmere length and cashmere length growth volume and decreased the cashmere fineness, with no statistical significance. The goat cashmere yield, goat weight after shearing, single and twin-birth kid weight and kids' mature secondary hair follicle density were significantly higher in the supplemented group (p < 0.05). In conclusion, supplementation in accordance with "Nutrient Requirements of Cashmere Goats" can enhance pregnant cashmere goats' fiber production, growth performance, fertility and kids' secondary hair follicles development, which is of great importance for the healthy and precise nutrition and management of cashmere goats.

Mesoporous waffle-like N-doped carbon with embedded Co nanoparticles for efficiently electrocatalytic oxygen reduction and evolution.

Rational design and facile preparation of high-performance carbon-based eletrocatalysts for both oxygen reduction and evolution reactions (ORR and OER) is crucial for practical applications of rechargeable zinc-air batteries. Inspired by the fact that the metallic Co catalysis on the formation of carbon nanotubes (CNTs), this work develops a facial compression-pyrolysis route to synthesize a mesoporous waffle-like N-doped carbon framework with embedded Co nanoparticles (Co@pNC) using a Co metal-organic framework and melamine as precursors. The unique porous waffle-like carbon framework is built up of interwoven N-doped CNTs and graphene nanosheets, which offers abundant catalytic-active sites and rapid diffusion channels for intermediates and electrolyte. The optimized Co@pNC shows excellent bifunctional ORR/OER electrocatalytic activity in alkaline media with a half-wave potential (E1/2) of 0.85 V for ORR and a small potential gap of 0.70 V between ORR E1/2 and OER potential at 10 mA cm-2. Its assembled battery exhibits a peak power density up to 150.3 mW cm-2, an energy density of 928 Wh kgZn-1 and superb rate capability. It highlights a facile component and architecture strategy to design high-performance carbon-based eletrocatalysts.

Steam explosion processing intensifies the nutritional values of most crop byproducts: Morphological structure, carbohydrate-protein fractions, and rumen fermentation profile.

To investigate the feasibility of steam explosion on the exploitation of ruminant feedstuff, the morphological structure, carbohydrate-protein fractions, and rumen fermentation profile of five typical crop byproducts (corn cob, rice straw, peanut shell, millet stalk, and sugarcane tip) were analyzed before and after steam explosion processing. The results showed that these crop byproducts had different physicochemical properties and rumen fermentation profiles, most of which could be improved by steam explosion processing, i.e., more rough morphological surface, much-broken structure, more digestible carbohydrate fraction (non-NDF +49.92-452.24%), faster gas production rate (c +9.72-68.75%), higher dry matter digestibility (DMD48 +11.38-47.36%), more available energy (ME -3.69-+42.13%, except for peanut shell), along with more unavailable protein fraction (ADICP +27.16-102.70%). It is suggested that steam explosion processing could intensify the feeding value of most crop byproducts for ruminants, but with a caution of heat damage to proteins.

Study on the Mechanism of Elastic Instability Caused by Natural Growth in Orthotropic Material.

Compared to synthetic materials, naturally grown biological materials have more specific behavioral patterns and life connotations in their morphological evolution over millions of years of environmental evolution on Earth. In this paper, we investigate the physical mechanisms and manifestations of out-of-plane deformation instability. Firstly, the origin of the instability phenomenon caused by the growth of the leaf is introduced. Leaf instability problems are modeled using rectangular thin plates. Secondly, the variation in the critical intrinsic strain with the principal shear modulus is obtained by numerical solution. The post-buckling behavior of the growth instability is further analyzed by general static analysis, and we obtain the phase diagram of morphogenesis of thin plant organs as functions of the principal shear modulus and off-axis angle. The research results enhance the understanding of the mechanism of elastic instability caused by natural growth in orthotropic materials.

Effect of tea polyphenols on the fermentation quality, protein preservation, antioxidant capacity and bacterial community of stylo silage.

The present study was aimed to evaluate the improvement potential of tea polyphenols (TPP) on silage characteristics and bacterial community. Stylo ensiled with TPP (0, 0.2 or 0.4%, on a fresh basis) were analyzed for fermentation parameter, protein fraction, antioxidant activity and bacterial community after 7, 14, 30 and 60 days fermentation. The addition of TPP resulted in the decrease (P < 0.05) of pH values (5.09 vs. 4.91), dry matter loss (11.77 vs. 8.02% DM), butyric acid concentration (1.64 vs. 1.02% DM) and ammonia-N proportion (13.69 vs. 8.98% CP, on Day 30) of stylo silage as well as the increase (P < 0.01) of lactic acid bacteria population (6.17 vs. 7.54 cfu/g FM) and true protein content (6.03 vs. 7.44% DM), particularly at the first 30 days of ensiling. It somewhat enhanced the antioxidant capacity of style silage at the early stage, and altered the bacterial community of stylo silage, with Clostridium_sensu_stricto_1 and Lachnoclostridium_5 much decreased but Enterobacter and Clostridium still being the dominant genera. It is suggested that TPP could help improve fermentation quality and nutrient preservation of stylo silage, and delay proteolysis process and antioxidant decay.

Validation of superior reference genes in mouse submandibular glands under developmental and functional regeneration states.

Saliva is crucial for lubricating the mouth and aiding in food digestion. However, the occurrence of oral dysfunction, such as xerostomia, dysphagia or oral infection can markedly reduce the quality of life of affected individuals. The major salivary glands include the submandibular gland (SMG), and sublingual and parotid glands; they are the larger glands in mammals that produce the majority of the saliva. The SMG serves as an effective model for the study of branching morphogenesis and functional regeneration. In order to better understand the key dynamic gene expression patterns during salivary gland development and functional regeneration, it is crucial to search for a panel of reliable reference genes. The present study thus aimed to identify superior reference genes to normalize gene expression data in the SMG under states of development and functional regeneration. First, the developmental SMG samples were harvested from mice in the embryonic and post­natal periods. Functional regeneration samples from a ductal ligation/de­ligation model were obtained at several stages. A total of 12 reference genes (Actb, Actg1, Ubc, Uba1, Uba52, Ube2c, Tuba1a, Tuba1b, Tubb5, H2afy, H2afx and Gapdh) from 430 candidates involving tubulin, histone, actin, ubiquitin and GAPDH family members were screened via transcriptome sequencing (RNA­seq) analysis. RT­qPCR (SYBR­Green) and western blot analysis were then used to semi­quantitatively assess gene and protein expression. The stability of expression was evaluated using the ΔCq, geNorm, BestKeeper, NormFinder and RefFinder methods and software. Actg1 exhibited the highest stability in the SMG developmental stage, while Tubb5 was recommended as the most stable reference gene for the SMG regenerative stage. In summary, the present study provides evidence­based selections for superior reference genes in the SMG during the stages of development and functional regeneration.