Integrated Analysis of the Transcriptome and Microbial Diversity in the Intestine of Miniature Pig Obesity Model.

Obesity, a key contributor to metabolic disorders, necessitates an in-depth understanding of its pathogenesis and prerequisites for prevention. Guangxi Bama miniature pig (GBM) offers an apt model for obesity-related studies. In this research, we used transcriptomics and 16S rRNA gene sequencing to discern the differentially expressed genes (DEGs) within intestinal (jejunum, ileum, and colon) tissues and variations in microbial communities in intestinal contents of GBM subjected to normal diets (ND) and high-fat, high-carbohydrate diets (HFHCD). After a feeding duration of 26 weeks, the HFHCD-fed experimental group demonstrated notable increases in backfat thickness, BMI, abnormal blood glucose metabolism, and blood lipid levels alongside the escalated serum expression of pro-inflammatory factors and a marked decline in intestinal health status when compared to the ND group. Transcriptomic analysis revealed a total of 1669 DEGs, of which 27 had similar differences in three intestinal segments across different groups, including five immune related genes: COL6A6, CYP1A1, EIF2AK2, NMI, and LGALS3B. Further, we found significant changes in the microbiota composition, with a significant decrease in beneficial bacterial populations within the HFHCD group. Finally, the results of integrated analysis of microbial diversity with transcriptomics show a positive link between certain microbial abundance (Solibacillus, norank_f__Saccharimonadaceae, Candidatus_Saccharimonas, and unclassified_f__Butyricicoccaceae) and changes in gene expression (COL6A6 and NMI). Overall, HFHCD appears to co-contribute to the initiation and progression of obesity in GBM by aggravating inflammatory responses, disrupting immune homeostasis, and creating imbalances in intestinal flora.

Responsive Peptide Nanofibers with Theranostic and Prognostic Capacity.

Photodynamic therapy (PDT) is a highly promising therapeutic modality for cancer treatment. The development of stimuli-responsive photosensitizer nanomaterials overcomes certain limitations in clinical PDT. Herein, we report the rational design of a highly sensitive PEGylated photosensitizer-peptide nanofiber (termed PHHPEG 6 NF) that selectively aggregates in the acidic tumor and lysosomal microenvironment. These nanofibers exhibit acid-induced enhanced singlet oxygen generation, cellular uptake, and PDT efficacy in vitro , as well as fast tumor accumulation, long-term tumor imaging capacity and effective PDT in vivo . Moreover, based on the prolonged presence of the fluorescent signal at the tumor site, we demonstrate that PHHPEG 6 NFs can also be applied for prognostic monitoring of the efficacy of PDT in vivo , which would potentially guide cancer treatment. Therefore, these multifunctional PHHPEG 6 NFs allow control over the entire PDT process, from visualization of photosensitizer accumulation, via actual PDT to the assessment of the efficacy of the treatment.

Vasorin Deletion in C57BL/6J Mice Induces Hepatocyte Autophagy through Glycogen-Mediated mTOR Regulation.

Abnormal vasorin (Vasn) expression occurs in multiple diseases, particularly liver cancers. Vasn knockout (KO) in mice causes malnutrition, a shortened life span, and decreased physiological functions. However, the causes and underlying mechanisms remain unknown. Here, we established Vasn KO C57BL/6J mice by using the CRISPR/Cas9 system. The animals were weighed, and histology, immunohistochemistry, electronic microscopy, and liver function tests were used to examine any change in the livers. Autophagy markers were detected by Western blotting. MicroRNA (miRNA) sequencing was performed on liver samples and analyses to study the signaling pathway altered by Vasn KO. Significant reductions in mice body and liver weight, accompanied by abnormal liver function, liver injury, and reduced glycogen accumulation in hepatocytes, were observed in the Vasn KO mice. The deficiency of Vasn also significantly increased the number of autophagosomes and the expression of LC3A/B-II/I but decreased SQSTM1/p62 levels in hepatocytes, suggesting aberrant activation of autophagy. Vasn deficiency inhibited glycogen-mediated mammalian target of rapamycin (mTOR) phosphorylation and activated Unc-51-like kinase 1 (ULK1) signaling, suggesting that Vasn deletion upregulates hepatocyte autophagy through the mTOR-ULK1 signaling pathway as a possible cause of diminished life span and health. Our results indicate that Vasn is required for the homeostasis of liver glycogen metabolism upstream of hepatocyte autophagy, suggesting research values for regulating Vasn in pathways related to liver physiology and functions. Overall, this study provides new insight into the role of Vasn in liver functionality.

Vasorin contributes to lung injury via FABP4-mediated inflammation.

BACKGROUND: Lung injury caused by pulmonary inflammation is one of the main manifestations of respiratory diseases. Vasorin (VASN) is a cell-surface glycoprotein encoded by the VASN gene and is expressed in the lungs of developing mouse foetuses. Previous research has revealed that VASN is associated with many diseases. However, its exact function in the lungs and the underlying mechanism remain poorly understood. METHODS AND RESULTS: To investigate the molecular mechanisms involved in lung disease caused by VASN deficiency, a VASN gene knockout (VASN-/-) model was established. The pathological changes in the lungs of VASN-/- mice were similar to those in a lung injury experimental mouse model. We further analysed the transcriptomes of the lungs of VASN-/- mice and wild-type mice. Genes in twenty-four signalling pathways were enriched in the lungs of VASN-/- mice, among which PPAR signalling pathway genes (3 genes, FABP4, Plin1, AdipoQ, were upregulated, while apoA5 was downregulated) were found to be closely related to lung injury. The most significantly changed lung injury-related gene, FABP4, was selected for further verification. The mRNA and protein levels of FABP4 were significantly increased in the lungs of VASN-/- mice, as were the mRNA and protein levels of the inflammatory factors IL-6, TNF-α and IL-1ß. CONCLUSIONS: We believe that these data provide molecular evidence for the regulatory role of VASN in inflammation in the context of lung injury.

Cavin-1 promotes M2 macrophages/microglia polarization via SOCS3.

PURPOSE: Our study aimed to investigate the function of Cavin-1 and SOCS3 in macrophages/microglia M2 polarization and further explored the relevant mechanism. METHODS: Expression levels of Cavin-1 and SOCS3 in macrophages/microglia were measured by western blotting and RT-PCR, respectively. Then, Cavin-1 or SOCS3 was gene silenced by a siRNA approach, and gene silencing efficiency was determined by western blotting. Next, co-immunoprecipitation (Co-IP) was employed to further analyze the interaction between Cavin-1 and SOCS3. Finally, the activation of STAT6/PPAR-γ signaling was evaluated using western blotting, and the M2 macrophages/microglia polarization was validated by measuring the mRNA expression of M2 markers by RT-PCR. RESULTS: In the polarization process of macrophages/microglia to M2 phenotype, both Cavin-1 and SOCS3 increased synchronously at protein and mRNA level, reached the peak at the 6 h, and then decreased. After Cavin-1 or SOCS3 silencing, the expression of Cavin-1 and SOCS3 declined. These results suggested that Cavin-1 and SOCS3 were positively correlated in macrophages/microglia, and this conjecture was verified by Co-IP. Besides, Cavin-1 silencing not only suppressed the activation of STAT6/PPAR-γ pathway, but also suppressed the release of anti-inflammatory factors. Finally, we found that SOCS3 overexpression reversed the inhibitory effect of Cavin-1 silencing on the release of anti-inflammatory factors in M2 macrophages/microglia. CONCLUSIONS: Cavin-1 and SOCS3 are actively involved in the process of M2 macrophages/microglia polarization. As a SOCS3 interacting protein, Cavin-1 can promote M2 macrophages/microglia polarization via SOCS3.

The pheromone affects reproductive physiology and behavior by regulating hormone in juvenile mice.

Pheromones could promote hormone secretions and regulate sexual behavior. It was unclear whether multiparous pheromone could induce variations in puberty. The aim was to ascertain whether pheromone in urine of multiparous females induced central precocious puberty (CPP) in juvenile C57BL/6J females. The precocious puberty was examined by vaginal smear, lordosis reaction, HE stain, and ELISA analysis. Results suggested that the first vaginal opening and the first estrus were significantly earlier. The time interval of the first vaginal opening and estrus was significantly shortened. It was interesting that the first estrus was significantly correlated with the first vaginal opening and the time interval of the first estrus. In the first estrus, female lordosis reaction, the number of mature follicles, and the weight of the ovary and uterus significantly increased. The level of luteinizing hormones also significantly increased. Thus, multiparous pheromone can regulate sex hormone to induce CPP in juvenile C57BL/6J females.

Effective Picosecond Nd:YAG laser on seborrheic dermatitis and its mechanism.

BACKGROUND: The Picosecond Nd:YAG laser has advantages in skin rejuvenation, which has little damage to surrounding tissues due to the ultra-short pulse width. We perform clinical application of Picosecond Nd:YAG laser's tender skin mode, which could improve the seborrheic dermatitis. MATERIALS AND METHODS: Sixty-three subjects with facial seborrheic dermatitis are randomized to be control and observation groups. Records regarding skin subjective improvement, skin barrier function-related data, skin microbial status, and dermoscopy detection of the two patient groups before and after treatments are investigated. RESULTS: Improvements of erythema and scales in observation group are significant compared with controls (p < 0.05). In terms of skin barrier function, there are significant changes regarding transepidermal water loss and epidermal seborrhea content in observation group after the laser treatments. Skin microbial state, pityrosporum furfur, and Demodex significantly decrement in observation group. Microscopical findings of infiltration mode advise that scales and capillary congestion and dilatation are significantly improved in observation group. Compared with controls, epidermal gloss increases, pore fineness improves, and capillary density decreases in the observation group. CONCLUSIONS: Picosecond Nd:YAG laser could efficiently decrease erythema area and seborrheic dermatitis scales and reduce pruritus incidence. The sebaceous gland secretion of seborrhea and multiplying of epidermal parasitic microbiological are inhibited after laser treatment. Less epidermal seborrhea content and inflammation are induced by parasitic microbiology, which is helpful for skin barrier function and microvascular remodeling.

Vasorin deficiency leads to cardiac hypertrophy by targeting MYL7 in young mice.

Vasorin (VASN) is an important transmembrane protein associated with development and disease. However, it is not clear whether the death of mice with VASN deficiency (VASN-/- ) is related to cardiac dysfunction. The aim of this research was to ascertain whether VASN induces pathological cardiac hypertrophy by targeting myosin light chain 7 (MYL7). VASN-/- mice were produced by CRISPR/Cas9 technology and inbreeding. PCR amplification, electrophoresis, real-time PCR and Western blotting were used to confirm VASN deficiency. Cardiac hypertrophy was examined by blood tests, histological analysis and real-time PCR, and key downstream factors were identified by RNA sequencing and real-time PCR. Western blotting, immunohistochemistry and electron microscopy analysis were used to confirm the downregulation of MYL7 production and cardiac structural changes. Our results showed that sudden death of VASN-/- mice occurred 21-28 days after birth. The obvious increases in cardiovascular risk, heart weight and myocardial volume and the upregulation of hypertrophy marker gene expression indicated that cardiac hypertrophy may be the cause of death in young VASN-/- mice. Transcriptome analysis revealed that VASN deficiency led to MYL7 downregulation, which induced myocardial structure abnormalities and disorders. Our results revealed a pathological phenomenon in which VASN deficiency may lead to cardiac hypertrophy by downregulating MYL7 production. However, more research is necessary to elucidate the underlying mechanism.

The Picosecond Laser Effects on Tattoo Removal and Metabolic Pathways.

OBJECTIVE: We aimed to investigate picosecond laser effects regarding rat tattoo removals. We systematically detected the metabolic pathways considering tattoo pigment particles in rat models. MATERIALS AND METHODS: We employed fluorescein to mark the tattoo dye and utilized a pattern needle to prepare tattoo on rat so as to establish the tattoo removal effects of picosecond laser methods employing animal models. We applied a picosecond laser to process the tattoo and observed the effects along with metabolic pathways for tattoo removal via pathology and imaging approaches. RESULTS: Based on the results of characterization, pathology and fluorescence, we discovered that the picosecond laser could effectively remove the pigment particles on rat skin, part of which would be metabolized out of the body through the circulatory system. After picosecond laser treatment, the fluorescence intensity of the rat skin engraved part was gradually weakened. At 8 h after the treatment, the metabolic organs had weak fluorescence, and there was no fluorescence in the metabolic organs at 12 h and 24 h after the treatment. CONCLUSION: The picosecond laser had perfect tattoo removal effects, which could be utilized as a reference for clinical tattoo removal.

Robust and nanostructured chitosan-silica hybrids for bone repair application.

In this study, chitosan-silica hybrids (CSHs) with superior mechanical strength and homogeneous dispersion of nano-sized silica particles were synthesized via a facile sol-gel method aiming for bone regeneration. The effects of varied acidic conditions of sol-gel reaction and inorganic/organic ratios on the performance of the hybrid were investigated. CSHs synthesized under weak acidic conditions (acetic acid, pH 4.0) showed a homogeneous nanostructure and robust strength (maximum compressive strength: 42.6 ± 3.3 MPa and 271 ± 31 MPa in wet and dry forms, respectively). However, those developed under the strong acidic condition (HCl, pH 4.0) and the strong acid condition plus lower pH (HCl, pH 2.8) tended to aggregate and exhibited inferior mechanical properties (compressive strength: 6.3 ± 0.3 MPa in wet form at pH 2.8). Under the latter conditions, the interactions between silica and chitosan were weak. Moreover, the mechanical properties of the CSHs could be tuned in a wide range by conveniently varying the inorganic/organic composition ratio between 50% and 70%. In vitro cytocompatibility study indicated that CSHs were non-cytotoxic. These results suggested that the weak acidic sol-gel process were essential for fabricating chitosan-silica hybrids with high mechanical strength, which had potential to be applied as a bone substitute.

The comparison of skin rejuvenation effects of vitamin A, fractional laser, and their combination on rat.

BACKGROUND: Because of long-term exposure of skin, skin aging is an unavoidable natural law with age. Traditional Vitamin A and novel ablative fractional laser technique both have the effects of skin rejuvenation, and studies have demonstrated both of them have apparent clinical efficacy and histology-improving effects on photo-aging skin. MATERIALS AND METHODS: 45 female healthy Wistar rats were selected and the depilation areas of every rat were divided into four regions: control region(Region A), Vitamin A acid region(Region B), combination treatment region(Region C), and fractional laser region(Region D). 0.025% Vitamin A acid cream was applied to Region B and C every day for 3 weeks; Region C and D were irradiated once with 10600nm CO2 fractional laser on the first day of the trail. The skin tissue was dissected and placed into liquid nitrogen according to the design. The real-time quantitative PCR and western blotting methods were taken to detect the expression changes of miR-29a, Akt, TGF-ß, and mRNA of type III pre-collagen. RESULTS: It can be seen from the results of the real-time quantitative PCR that the mRNA expression levels of type III pre-collagen, Akt, and TGF-ß in the treatment regions are up-regulated and the expression levels of miR-29a mRNA are down-regulated compared to the Region A. The hybridization tests showed that changes of the expression of type III pre-collagen, Akt gene, miR-29a gene, and TGF-ß gene across the experiment regions are all significantly different in the third week, and the expression levels of them all achieve the highest value in the third week, the expression level of miR-29a gene achieves the lowest value in the third week, which are consistent with the results of real-time quantitative PCR. CONCLUSION: It is indicated that the combination region of Vitamin A acid and fractional laser may lead to low expression of miR-29a, thus the inhibition of downstream Akt activation is loss, Akt activation is enhanced, enhancement of the expression of TGF-ß is induced, leading to proliferation of fibroblasts, and promotion of the collagen proteins' synthesis in skin. Therefore miR-29a/Akt/TGF-ß signal pathway may participate in the skin rejuvenation mechanism of action Vitamin A acid and fractional laser. This may provide a new treatment approach for skin rejuvenation.

miRNA-126 enhances viability, colony formation, and migration of keratinocytes HaCaT cells by regulating PI3 K/AKT signaling pathway.

Wound healing is a basic biological process including proliferation and migration of keratinocyte. The effects of microRNAs on skin wound healing remain largely unexplored. This study aimed to investigate the role of microRNA-126 (miR-126) in human skin wound healing. Relative expression of miR-126 after injury was evaluated by qRT-PCR. Cell viability, colony formation, cycle distribution, migration, and the alternation of PI3 K/AKT pathway after miR-126 knockdown or overexpression were detected, respectively. In addition, potential target gene of miR-126 was also explored by luciferase assay. Results showed that miR-126 was up-regulated during skin wound healing. Moreover, overexpression of miR-126 promoted cell proliferation and migration, whereas inhibition of miR-126 led to the opposite effects. Additionally, we discovered that PLK2, which inhibited cell viability, colony formation and migration of keratinocyte, was a target gene of miR-126. The expression of PLK2 was negatively correlated with the level of miR-126 during wound healing. Finally, we demonstrated that overexpression of miR-126 significantly increased the expression of p-AKT, p-ERK2, and PI3 K, indicating that overexpression of miR-126 activated PI3 K/AKT signaling pathway. In conclusion, our results demonstrated that miR-126 acted as a critical regulator for promoting proliferation and migration in keratinocyte during skin wound healing.

Fabrication of novel bioactive hydroxyapatite-chitosan-silica hybrid scaffolds: Combined the sol-gel method with 3D plotting technique.

Sol-gel derived organic/inorganic hybrids, in which organic and inorganic components form co-networks at the molecular level, have demonstrated great potential for providing improved mechanical properties and biological functions in tissue engineering applications. Here, a novel bioactive hydroxyapatite-chitosan-silica hybrid (HA-CSH) scaffold was successfully fabricated by combining the sol-gel method and 3D plotting technique. Physiochemical characterization confirmed that chitosan was hybridized homogeneously with the inorganic phase on nanoscale. The obtained scaffolds possessed precisely controllable and interconnected porous structures. The nano-sized HA formed in situ and dispersed uniformly in the hybrid network, which reduced the water absorption and increased the mechanical strength of the hybrid scaffold under humidity condition as compared to chitosan-silica hybrid (CSH) scaffold. Compression tests showed that the 3D plotted hybrid scaffolds under wet conditions had compressive strengths of 10-13 MPa and elastic moduli of 21-27 MPa and thus met the mechanical requirements of human trabecular bone. Studies on the mineralization process under simulated body fluid (SBF) conditions confirmed that the introduction of HA obviously increased the biological activity of hybrid scaffolds. In vitro cell results indicated that the HA-CSH scaffold not only supported adhesion and proliferation of mouse bone mesenchymal stem cells (mBMSCs), but also improved the osteoinductivity. The alkaline phosphatase activity and mineral deposition on the HA-CSH scaffold were higher than those on the CSH scaffold. These results suggested that the 3D plotted HA-CSH scaffold may be a promising bioactive material for bone tissues regeneration.

Tea polyphenols inhibit the proliferation, migration, and invasion of melanoma cells through the down-regulation of TLR4.

Melanoma is the most common skin cancer and malignant melanoma which can cause skin cancer-related deaths. Toll-like receptor 4 (TLR4) had been reported to play an important role in melanoma, and tea polyphenol (TP) is regarded as an anticancer substance. However, the relationship between TP and TLR4 in melanoma is not well explored. Therefore, our aim is to figure out how TP has an influence on melanoma. Melanoma cell lines (B16F10 and A375) were treated with TP and lipopolysaccharides (LPS). Western blot assay was used to examine TLR4 expression, and MTT assay was conducted to assess proliferation. Wound healing assay was conducted to evaluate the migration of melanoma cells, and transwell assay was used to examine the melanoma cells' invasiveness. Besides, in vivo experiments were practiced for TP function in mice with melanoma cells. TP inhibited the proliferation, migration and invasion ability of melanoma cells, which displayed a dosage and time dependence. TLR4 was highly expressed in melanoma cells compared with normal skin cells. TP could suppress TLR4 expression both in normal melanomas and in stimulated melanomas by TLR4 agonist LPS. Suppressing TLR4 in melanomas could inhibit cell function (proliferation, migration, and invasion), and blocking the expression of 67LR could abolish TP function on TLR4. TP can inhibit melanoma (B16F10) growth in vivo.