[Effect of Uremic Clearance Granules on improvement of chronic kidney disease in rats based on microbiome-metabolomics and its mechanism].

This research aimed to study the effect of Uremic Clearance Granules on chronic kidney disease in SD rats by using the methods of microbial functional genomics combined with metabolomics, and to preliminarily explore its mechanism. The SD rat model of chronic kidney disease was established by the adenine-induced method. After the model was successfully induced, the animals were randomly divided into a negative control group, a Uremic Clearance Granule treatment group, and a normal control group, with 8 rats in each group. After 4 weeks of administration, animal feces and serum were collected, and 16S rDNA sequencing technology was used to analyze the abundance, diversity, and function prediction of intestinal microorganisms. Liquid chromatography-mass spectrometry(LC-MS) technology was used to perform high-throughput sequencing to detect animal serum metabolites. The MetPA database was used to screen out potential biomarkers of chronic kidney disease in rats and conduct the enrichment analysis of metabolic pathways. Spearman's method was used to analyze the correlation between the two omics. The results showed that Uremic Clearance Granules effectively improved the body weight loss and renal function-related biochemical and appearance indicators in rats with chronic kidney disease. The results of 16S rDNA sequencing showed that Uremic Clearance Granules regulated the diversity and composition of the intestinal flora in rats with chronic kidney disease. The changes in the intestinal flora affected functional metabolic pathways such as amino acid biosynthesis and metabolism, lipid metabolism, and carbohydrate metabolism. The results of LC-MS showed that as compared with the negative control group, 15 metabolites were reversed in the Uremic Clearance Granule treatment group, among which 11 potential marker metabolites were significantly up-regulated and 4 potential marker metabolites were significantly down-regulated. Five amino acid metabolic pathways were mainly involved, which were significantly correlated with changes in the intestinal flora. Therefore, Uremic Clearance Granules can improve the renal function of rats with chronic kidney disease, and the mechanism may be related to its effect on the amino acid metabolism pathway by regulating the intestinal flora.

Mutual regulation between miR-21 and the TGFß/Smad signaling pathway in human bronchial fibroblasts promotes airway remodeling.

OBJECTIVE: Airway remodeling is an important pathological feature of asthma. Excessive deposition of extracellular matrix (e.g., collagen) secreted from fibroblasts is a major factor contributing to airway remodeling. Currently, the mechanism by which collagen continues to be oversynthesized in the airway remains unclear. In this study, we investigated the role of the microRNA-21 (miR-21) and TGFß/Smad signaling pathway in human bronchial fibroblasts (HBFs), and explored the regulatory mechanism of airway remodeling. METHODS: HBFs were cultured in vitro and treated with the transforming growth factor ß (TGFß), receptor inhibitor (SB431542), and TGFß1. miR-21 and Smad7 overexpressing lentiviruses, as well as an miR-21 interfering lentivirus were constructed and transfected into HBFs. Western blotting was used to determine the expression of airway remodeling-related proteins and proteins in the TGFß/Smad signaling pathway. miR-21 expression was measured by quantitative real-time PCR. RESULTS: The high expression of miR-21 induced by TGFß1 was reduced following the treatment with the SB431542 in HBFs. Smad7 overexpression inhibited the elevated expression of the COL I protein induced by miR-21 overexpression in HBFs. Inhibiting miR-21 expression upregulated the level of Smad7 protein, thus reducing the expression of airway remodeling-related proteins induced by TGFß1 stimulation in HBFs. CONCLUSIONS: TGFß1 can induce miR-21 expression in HBFs through the TGFß/Smad signaling pathway to promote airway remodeling. miR-21 downregulates Smad7, activates the TGFß/Smad signaling pathway, and promotes airway remodeling. Mutual regulation between miR-21 and the TGFß/Smad signaling pathway in HBFs promotes airway remodeling.

[Primary study on collagen X gene expression in the apical disc of idiopathic scoliosis].

OBJECTIVE: To study the distribution of collagen X in the intervertebral disc and determine its role in the pathogeny of idiopathic scoliosis (IS). METHODS: The data included apical disc and intermediate disc from 14 cases of AIS, 26 discs from 13 cases of scoliosis of confirmed pathogeny which included 10 cases of congenital scoliosis and neurofibromatosis scoliosis (CS group). Six discs were obtained from 3 cases of sudden death of normal young man served as controls. The distribution of collagen X in the apical disc of IS was examined by immunohistochemistry and in situ hybridization (ISH) with RNA probe. The figure of collagen X hybridization in the endplate cartilage was input to the figure analysis system. The mRNA content of collagen X was compared between every 2 groups by SPSS software. RESULTS: Collagen X was mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its mRNA was expressed in the hypertrophic chondrocyte. Positive signal was found in the nuclei of several scoliosis patients, which was not related to the pathogeny. The collagen X mRNA contents of the apical disc and intermediated disc of the IS group and of the CS group were significantly higher than those of the normal group (P < 0.05). There was no difference between the IS and CS groups. CONCLUSIONS: Collagen X is mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its can also be secreted by the chondrocyte-like cells in the nucleus under special condition. Higher expression of collagn X gene in scoliosis patients may be the effects of long term abnormal stress which causes calcification of endplate cartilage. Collagen X expressed in the nucleus may be the result of secretion of chondrocyte-like cells in the disc under abnormal mechanical condition.

Temporal effect of local hyperthermia on murine contact hypersensitivity.

BACKGROUND: The sensitization and elicitation phases are involved in the immunopathogenesis of contact hypersensitivity (CHS). Langerhans cells (LCs) are believed to play pivotal roles in the sensitization stage of CHS. Local hyperthermia on skin induces the migration as well as maturation of epidermal LCs. Although fever-range whole body hyperthermia and local hyperthermia at 43°C prior to sensitization were reported to suppress CHS, the effects of different temperatures and the timing sequence of local hyperthermia on CHS have not been tackled. METHODS: Local hyperthermia was applied to murine dorsal skin 3 days prior to, concurrent with, or 2 days post sensitization with fluorescein isothiocyanate (FITC) in BALB/c mice. Local hyperthermia temperatures at 37°C, 39°C, 41°C and 43°C were applied to mouse dorsal skin and the severity of CHS was calculated by measuring the swelling response of the challenged ears. RESULTS: Local hyperthermia at 39°C, 41°C and 43°C prior to sensitization reduced the severity of CHS, as compared with that at 37°C. The suppression of CHS was temperature dependent in that higher temperature had a stronger effect. On the contrary, the hyperthermia treatments, either concurrent with or post-sensitization, resulted in an enhanced temperature-dependent ear swelling response. CONCLUSIONS: The severity of murine CHS could be influenced by local hyperthermia at the sensitization stage in a temperature dependent manner. The temporal effect of local hyperthermia suggested a novel factor in interpreting the severity of allergic contact dermatitis.