Sarsasapogenin restores podocyte autophagy in diabetic nephropathy by targeting GSK3ß signaling pathway.

Podocyte injury following abnormal podocyte autophagy plays an indispensable role in diabetic nephropathy (DN), therefore, restoration of podocyte autophagy is considered as a feasible strategy for the treatment of DN. Here, we investigated the preventive effects of sarsasapogenin (Sar), the main active ingredient in Anemarrhena asphodeloides Bunge, on the podocyte injury in diabetic rats, and tried to illustrate the mechanisms underlying the effects in high glucose (HG, 40 mM)-treated podocytes (MPs). Diabetes model was established in rats with single streptozocin (60 mg· kg-1) intraperitoneal administration. The rats were then treated with Sar (20, 60 mg· kg-1· d-1, i.g.) or a positive control drug insulin (INS) (40 U· kg-1· d-1, i.h.) for 10 weeks. Our results showed that both Sar and insulin precluded the decreases of autophagy-related proteins (ATG5, Beclin1 and LC3B) and podocyte marker proteins (podocin, nephrin and synaptopodin) in the diabetic kidney. Furthermore, network pharmacology was utilized to assess GSK3ß as the potential target involved in the action of Sar on DN and were substantiated by significant changes of GSK3ß signaling in the diabetic kidney. The underlying protection mechanisms of Sar were explored in HG-treated MPs. Sar (20, 40 µM) or insulin (50 mU/L) significantly increased the expression of autophagy- related proteins and podocyte marker proteins in HG-treated MPs. Furthermore, Sar or insulin treatment efficiently regulatedphosphorylation at activation and inhibition sites of GSK3ß. To sum up, this study certifies that Sar meliorates experimental DN through targeting GSK3ß signaling pathway and restoring podocyte autophagy.

Separate base usages of genes located on the leading and lagging strands in Chlamydia muridarum revealed by the Z curve method.

BACKGROUND: The nucleotide compositional asymmetry between the leading and lagging strands in bacterial genomes has been the subject of intensive study in the past few years. It is interesting to mention that almost all bacterial genomes exhibit the same kind of base asymmetry. This work aims to investigate the strand biases in Chlamydia muridarum genome and show the potential of the Z curve method for quantitatively differentiating genes on the leading and lagging strands. RESULTS: The occurrence frequencies of bases of protein-coding genes in C. muridarum genome were analyzed by the Z curve method. It was found that genes located on the two strands of replication have distinct base usages in C. muridarum genome. According to their positions in the 9-D space spanned by the variables u1 - u9 of the Z curve method, K-means clustering algorithm can assign about 94% of genes to the correct strands, which is a few percent higher than those correctly classified by K-means based on the RSCU. The base usage and codon usage analyses show that genes on the leading strand have more G than C and more T than A, particularly at the third codon position. For genes on the lagging strand the biases is reverse. The y component of the Z curves for the complete chromosome sequences show that the excess of G over C and T over A are more remarkable in C. muridarum genome than in other bacterial genomes without separating base and/or codon usages. Furthermore, for the genomes of Borrelia burgdorferi, Treponema pallidum, Chlamydia muridarum and Chlamydia trachomatis, in which distinct base and/or codon usages have been observed, closer phylogenetic distance is found compared with other bacterial genomes. CONCLUSION: The nature of the strand biases of base composition in C. muridarum is similar to that in most other bacterial genomes. However, the base composition asymmetry between the leading and lagging strands in C. muridarum is more significant than that in other bacteria. It's supposed that the remarkable strand biases of G/C and T/A are responsible for the appearance of separate base or codon usages in C. muridarum. On the other hand, the closer phylogenetic distance among the four bacterial genomes with separate base and/or codon usages is necessary rather than occasional. It's also shown that the Z curve method may be more sensitive than RSCU when being used to quantitatively analyze DNA sequences.

Re-prediction of protein-coding genes in the genome of Amsacta moorei entomopoxvirus.

Using the Z curve method, the protein-coding genes in AmEPV genome are re-predicted. On the basis of the parameters trained on the experimentally validated genes, all of the 30 experimentally validated genes and 67 putative genes are predicted correctly as coding genes. The sensitivities of the present method for self-test and cross-validation are all 100% based on these test sets. Thirty-eight annotated conserved and hypothetical genes are predicted as non-coding ORFs. The number of re-predicted protein-coding genes in AmEPV is 256. It is significantly less than the number 294 reported in the original annotation. After extending the present method trained in AeEPV genome to the other entomopoxvirus genome, it is found that 116 of the 123 known and putative genes are predicted correctly as coding. Six of the seven falsely missed genes are less than 300bp. The present method could be extended to other poxvirus genomes with or without adaptation of training sets.

Mechanism study of electrochemical oxidation in the terylene diaphragm cell.

By using a self-made carbon/polytetrafluoroethylene (C/PTFE) O2-fed as cathode and Ti/IrO2/RuO2 as anode, the effects of electrochemical oxidation of phenol and the coal-gas wastewater containing phenol were studied. The terylene diaphragm which kept pH >12 in cathodic compartment and pH < 1 in the anodic compartment was selected in the experiment in comparison with the other types of diaphragm. Furthermore, hydroxyl radical (HO*) was determined in the cathodic compartment of the diaphragm cell by electron spin resonance spectrum (ESR) and the fluorescence spectra. Compared with pH, the accumulated H2O2 and the COD removal of the no-diaphragm cell, the mechanism of electrochemical oxidation in the terylene diaphragm cell was supposed. The degradation of phenol was supposed to be cooperative oxidation by direct or indirect electrochemical oxidation at the anode and H2O2, HO* produced by oxygen reduction at the cathode. The mineralization of phenol in the diaphragm cell was better than that in the no-diaphragm cell. When the coal-gas wastewater was treated by the electrolysis system with terylene diaphragm, the average removal efficiency of the volatile phenol and COD were 100% and 79.6%, respectively.

Spatial variation and subcellular binding of metals in oysters from a large estuary in China.

Pearl River Estuary (PRE) is the largest estuary in Southern China and there has been an increasing concern of metal pollution due to regional industrialization. In this study, we investigated the spatial variation of metal pollution (Ag, As, Cd, Cu, Pb, and Zn) as well as their subcellular handling in the oyster Crassostrea hongkongensis. Hot spots of metal contamination in the oysters were found in different sites, suggesting that there were different sources of metals in the estuary associated with industrial activity. Metals differed in their subcellular bindings in the oysters from different locations. Metal distribution in the biologically detoxified fraction decreased for Cu but increased for Zn with increasing contamination in the oysters. For Zn, there was a significant difference in its two detoxification pools (metal-rich granules and metallothionein-like proteins) in response to Zn contamination. The high Cd concentrations in oysters may carry a high Cd hazard to the consumers.