The Intestinal Microbiota Composition in Early and Late Stages of Diabetic Kidney Disease.

Many studies have suggested that gut microbiota dysbiosis may be one of the pathogenesis factors of diabetes mellitus (DM), while it is not clear whether it is involved in the development of diabetic kidney diseases (DKD). The objective of this study was to determine bacterial taxa biomarkers during the progression of DKD by investigating bacterial compositional changes in early and late DKD. 16S rRNA gene sequencing was performed on fecal samples, including the diabetes mellitus (DM), DNa (early DKD), and DNb (late DKD) groups. Taxonomic annotation of microbial composition was performed. Samples were sequenced on the Illumina NovaSeq platform. At the genus level, we found counts of Fusobacterium, Parabacteroides, and Ruminococcus_gnavus were significantly elevated both in the DNa group (P = 0.0001, 0.0007, and 0.0174, respectively) and the DNb group (P < 0.0001, 0.0012, and 0.0003, respectively) compared with those in the DM group. Only the level of Agathobacter was significantly decreased in the DNa group than the DM group and in the DNb group than the DNa group. Counts of Prevotella_9, Roseburia were significantly decreased in the DNa group compared with those in the DM group (P = 0.001 and 0.006, respectively) and in the DNb group compared with those in the DM group (P < 0.0001 and 0.003, respectively). Levels of Agathobacter, Prevotella_9, Lachnospira, and Roseburia were positively correlated with an estimated glomerular filtration rate (eGFR), but negatively correlated with microalbuminuria (MAU), 24 h urinary protein quantity (24hUP), and serum creatinine (Scr). Moreover, the areas under the curve (AUCs) of Agathobacter and Fusobacteria were 83.33% and 80.77%, respectively, for the DM and DNa cohorts, respectively. Notably, the largest AUC for DNa and DNb cohorts was also that of Agathobacter at 83.60%. Gut microbiota dysbiosis was found in the early and late stages of DKD, especially in the early stage. Agathobacter may be the most promising intestinal bacteria biomarker that can help distinguish different stages of DKD. IMPORTANCE It is not clear as to whether gut microbiota dysbiosis is involved in the progression of DKD. This study may be the first to explore gut microbiota compositional changes in diabetes, early-DKD, and late DKD. We identify different gut microbial characteristics during different stages of DKD. Gut microbiota dysbiosis is found in the early and late stages of DKD. Agathobacter may be the most promising intestinal bacteria biomarker that can help distinguish different stages of DKD, although further studies are warranted to illustrate these mechanisms.

Carthamin Yellow Protects the Heart Against Ischemia/Reperfusion Injury With Reduced Reactive Oxygen Species Release and Inflammatory Response.

Carthamin yellow (CY) is a flavonoid compound isolated from safflower, which is widely used clinically in China. It has various pharmacological effects including promoting blood circulation to remove blood stasis and alleviating pain. Ischemic heart disease is one of the main culprits of illness and death. Here, in this study, ex vivo and in vivo models were used to investigate whether CY reduces ischemia/reperfusion injury. In vitro experiments further verify and explain the potential mechanisms of CY cardioprotective function. Isolated hearts from male rats with or without CY pretreatment before ischemia which underwent 30-minute ischemia followed by 60-minute reperfusion showed that CY pretreatment significantly reduced the infarct size and lactate dehydrogenase release. The in vivo experiments also indicated CY preadministration (i.v.) reduced infarct size and improved the heart function, which was impaired by myocardial ischemia/reperfusion injury. The in vitro model on myocardial cell also showed that CY reduced ischemia/reperfusion injury by reducing the lactate dehydrogenase and reactive oxygen species (ROS) releasing. Eliminating ROS with N-acetylcysteine or preinject CY into rat jugular vein reduces the expression of IL-6, TNF-a, and, especially, IL-1b in an in vivo I/R model. Also, CY pretreatment strongly reduces ischemia/reperfusion-induced NLRP3 up-expression and caspase-1 activation. Our results indicated CY reduced ischemia-reperfusion injury when administered before reperfusion. The reduction in injury is accompanied by a reduced ROS release and decreased inflammatory response.

[Preparation of rat model of systemic inflammatory response syndrome induced by zymosan].

OBJECTIVE: To establish a model of systemic inflammatory response syndrome (SIRS) in rats. METHODS: SD rats were intraperitoneally injected with different concentrations of zymosan suspension. The general status, temperature, white cell count, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-10 (IL-10) and the pathological changes of main organs were examined. RESULTS: The conditions of rats receiving zymosan doses of 750 mg/kg and 1000 mg/kg were consistent with the criteria of SIRS model; however, the mortality of 1000 mg/kg group was higher than that of 750 mg/kg group. CONCLUSION: The rat model of systemic inflammatory response syndrome has been successfully induced.

[Transfection of pancreatic cancer cells BxPC-3 with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA and its silencing effects].

OBJECTIVE: To study the transfection of pancreatic cancer cells BxPC-3 with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA and its silencing effects. METHODS: Pancreatic cancer cells BxPC-3 transfected with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA were selected as target and divided into five groups: (1) T1 group (pSilencer4.1CMV neo-hTERT1-siRNA), (2) T2 group (pSilencer4.1CMV neo-hTERT2-siRNA), (3) Lipofectamine (Lipofectamine), (4) mismatch group(pSilence4.1CMV, as negative control), (5) cell control group(without transfection). The expression of hTERT mRNA was detected by RT-PCR. The viability of cells was measured by MTT method. The cell cycle and cell apoptosis was measured by flow cytometry. The expression of telomerase protein was measured by Western blot. RESULTS: Compared with Lipofectamine group, negative control group and cell control group, the expression of hTERT-mRNA and telomerase protein in cells was downregulated significantly(P<0.05), the viability of BxPC-3 cells was decreased significantly (P<0.05), the ratio of cells in G0/G1 stage was increased, the ratio of cells in S stage and G2/M stage was decreased, and the ratio of apoptotic cells was increased significantly in T1 group and T2 group. CONCLUSION: Recombinant plasmid T1 and T2 can downregulate the expression of hTERT mRNA and telomerase protein in BxPC-3 cells , and has good RNAi silencing effects. T1 and T2 can also inhibit the growth of BxPC-3 cells, block the cell cycle, promote the apoptosis of cells, and has anti-pancreatic cancer effects in vitro.