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Methods: Blood pressure and urine biochemical indices were recorded. Renal blood flow was evaluated by renal ultrasonography. Transmission electron microscopy (TEM) and HE staining were used to assess kidney and spleen morphology. Renal fibrosis was assessed using Masson staining. Serum levels of IL-6, IL-10, and IL-17A were measured using ELISAs. The density of RORγ and Foxp3 in the spleen was observed by immunofluorescence staining. The levels of Th17 cells and Tregs in blood were detected via flow cytometry. Transcriptome sequencing was performed to screen the targets of BSHM granules in hypertensive kidneys. Results: BSHM granules decreased SBP by 21.2 mm·Hg and DBP by 8.8 mm·Hg in ageing SHRs (P < 0.05), decreased the levels of urine mALB, ß2-Mg, and NAG (P < 0.01), and improved renal blood flow and arteriosclerosis. BSHM granules increased IL-10 expression (P < 0.05) while decreasing IL-6 (P < 0.01) and IL-17A (P < 0.05) levels. BSHM granules improved Foxp3 density and the number of Tregs (P < 0.01) and reduced RORγt density and the number of Th17 cells (P < 0.01). Transcriptome sequencing identified 747 differentially expressed (DE) mRNAs in kidneys after BSHM treatment. GO analysis suggested that BSHM granules act through immunoregulation. Conclusions: BSHM granules attenuated hypertensive renal damage in ageing SHRs, by significantly increasing Tregs and decreasing Th17 cells.
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Using a series of DNA methylation analysis, pathogenesis was investigated to identify the specific DNA methylation markers for diagnosing atherosclerosis. Firstly, with the chip platform of Illumina Human Methylation 450 BeadChip, a total of 1,458 CpGs, covering 971 differential methylated genes were extracted with stringent filtering criteria. Secondly, hierarchical clustering as a heat map was used to check on the dependability of differential methylated genes. Thirdly, the related GO terms and pathways were enriched by up- and down-methylated genes, respectively, after verifying the capacity of these differential methylated genes to distinguish between atherosclerosis and healthy controls. In total, 971 differential DNA methylated genes were identified (1,458 CpGs). Several important function regions were also identified, including cell adhesion, PI3K-Akt signaling pathway and transcription from RNA polymerase II promoter. This study indicates that patients with atherosclerosis have high levels of DNA methylation, which is promising for early diagnosis and treatment of atherosclerosis.
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In this study, we evaluated the effectiveness of SrCl2·6H2O to mark otoliths of juvenile common carp (Cyprinus carpio). The carp fish were immersed with four concentrations of the Sr compound (i.e., 0, 4, 8, 12 mg·L-1) for 2 d to determine the essential concentration for immersing. Then, the carp fish were immersed in 8 mg·L-1 of the Sr compound for five different immersing times (i.e., 1, 2, 3, 4, 5 d) to establish the essential immersing time. Results from electron microprobe analysis (EPMA) showed that otolith Sr marks were very obvious in all C. carpio fish of the aforementioned Sr immersed group with high ratio of Sr/Ca concentration and red color (high Sr level) ring map, compared to the control group with low ratio of Sr/Ca concentration and basic blue color (low Sr level) map. We obtained a 100% mark-success rate of otolith Sr marking in juvenile C. carpio. No significant difference were observed on the average body length, mass and mortality between the immersed group and the control group, suggesting that Sr marking had no negative impact on fish. As clear and complete otolith Sr marked rings occurred at concentration of 8 mg·L-1 or more, and immersing time of 2 d or more for the Sr compound, 8 mg·L-1 and 2 d were suggested as the essential immersing Sr concentration and immersing time, respectively. Our results demonstrated the strong feasibility of otolith Sr marking for juvenile C. carpio.