[Twenty years in the 21st century: identification technologies for and multimodal research on chemical changes in exploration of processing mechanism of Chinese medicine].

Chinese medicine undergoes complex chemical changes during processing and identifying these changes is the key to the processing mechanism. In the past 20 years of the 21 st century, research on the chemical changes in Chinese medicine after processing has focused the changes in the biopharmaceutical process in addition to the variation during processing. With the surging of information technologies, various identification technologies(instrumental analysis techniques, molecular biological techniques, data mining techniques, and biotransformation techniques) have developed rapidly and been widely applied to the research on processing mechanism. Thus, based on the chemical changes in the processing and biopharmaceutical process, the author suggested a research tactic of multimodal identification as the core by reorganizing key technologies for chemical identification from studies of the processing mechanism of Chinese me-dicine, aiming at establishing an interdisciplinary multi-dimensional research model for the processing mechanism of Chinese medicine.

[Twenty years in the 21st century: research approaches and techniques in modern system biology for mechanisms of Chinese medicinal processing].

Clarifying the mechanisms of Chinese medicinal processing is pivotal to the modernization of Chinese medicine. Research on Chinese medicinal processing gives priority to the mechanisms of the processing in enhancing efficacy, reducing toxicity, and repurposing medicinals. During the past 20 years, scholars have carried out in-depth studies on the mechanisms of Chinese medicinal processing via modern system biology. They mainly focused on the changes of medicinal properties and efficacy caused by processing using techniques of modern pharmacology and molecular biology, spectrum-efficacy correlation, and biophoton emission. However, these techniques fail to reflect the holistic view of traditional Chinese medicine. With the introduction of system biology, multi-omics techno-logies(genomics, transcriptomics, proteomics, and metabolomics) have surged, which have been applied to the research on the mec-hanisms of Chinese medicinal processing. These multi-omics technologies have advantages in the research on holism. This study aims to summarize the research techniques and approaches in system biology for mechanisms of Chinese medicinal processing in the past 20 years and analyze the limitations and advantages of them. It is concluded that the multi-omics techniques of system biology can reconstruct the mechanisms of Chinese medicinal processing. This study provides a new direction for further research on the mechanisms of Chinese medicinal processing.

[Gaseous Nitrogen Emission from Soil After Application of NH4+-N Loaded Biochar].

To safely and effectively transfer NH4+-N from eutrophic water to soil, biochar was applied to adsorb NH4+-N from wastewater, and this NH4+-N loaded biochar (N-BC) was subsequently used as a soil amendment. Understanding the influence of N-BC on N2O-N emission and NH3-N volatilization is important for both decreasing the application of chemical fertilizers and reducing gaseous nitrogen loss from soil. In this study, experiments were conducted in soil columns with four treatments, namely CK (no fertilizer), NPK (chemical fertilizer), N-BC+PK (NH4+-N loaded biochar+chemical fertilizer), and BC+NPK (biochar+chemical fertilizer). Compared to both the NPK and BC+NPK treatments, N-BC+PK significantly reduced the cumulative N2O-N emissions and NH3-N volatilization, as well as the total gaseous nitrogen loss from the soil (P<0.05). Relative to NPK and BC+NPK, cumulative N2O-N emissions decreased by 33.62% and 24.64%, cumulative NH3-N volatilization decreased 70.64% and 79.29%, and the cumulative total gaseous nitrogen loss decreased by 64.97% and 73.75%. In particular, BC+NPK significantly enhanced the cumulative NH3-N volatilization. Furthermore, the N2O-N emission flux and NH3-N volatilization rate were significantly positively correlated with the NH4+-N concentration, NO3--N concentration, and pH of soil (P<0.01). Overall, using NH4+-N loaded biochar can significantly decrease N2O-N emissions and NH3-N volatilization, relative to the traditional application combining biochar and chemical fertilizer. This research provides solid theoretical support and data for the application of NH4+-N loaded biochar in soil, to reduce gaseous nitrogen loss.

[Effect of Applying Hydrochar for Reduction of Ammonia Volatilization and Mechanisms in Paddy Soil].

Hydrochar, as a product of the hydrothermal carbonization of biomass, has good application prospects for the NH3 volatilization reduction in rice fields due to its rich pore structure and functional surface. In this study, hydrochar was applied as a soil conditioner to paddy soil. A soil column experiment was conducted to investigate the effect of hydrochar on NH3 volatilization throughout the growth period of rice. The experiment was conducted with three treatments:CKU (control without hydrochar); SHC (sawdust hydrochar); and W-SHC (water-washed sawdust hydrochar). The application rate of SHC and W-SHC was 0.5% (w/w). The study investigated the effects of different hydrochars on the pH and concentrations of NH4+-N in floodwater, the flux and accumulation of NH3 volatilization, and the yield-scale cumulative emission of NH3 volatilization. Results show that the SHC treatment significantly reduces cumulative emissions of NH3 volatilization and the yield-scale cumulative emissions of NH3 volatilization (P<0.05), which were 32.42% and 47.61% lower than CKU, respectively. The effect of W-SHC on ammonia volatilization reduction was slightly weaker, as the cumulative emissions of NH3 volatilization and the yield-scale cumulative emissions of NH3 volatilization decreased by 10.14% and 27.71%, respectively, compared with CKU. The NH3 volatilization reduction was possibly related to the disturbance of pH and the decrease in NH4+-N concentrations in the floodwater because of the application of hydrochar. Compared with CKU, both SHC and W-SHC treatments reduced the pH and NH4+-N concentration in the floodwater. The impacts were more obvious in the rice base fertilizer period (BF) and the first supplemental fertilizer period (SF1) than in the second supplemental fertilizer period (SF2). The soil urease activity was significantly inhibited by hydrochar (P<0.05), and the abundance of soil ammonia-oxidizing gene (AOA, AOB) also significantly increased after application of SHC (P<0.05). This resulted in the enhanced efficiency of ammonia-oxidizing, which had an effect on the reduction of the NH4+-N concentrations in the floodwater. This study provides theoretical and experimental data support for the application of hydrochar in agro-environments with regard to ammonia volatilization reduction in paddy fields.

Cathepsin D protects renal tubular cells from damage induced by high glucose independent of its enzymatic activity.

Although glomerular and vascular damage have been considered the main characteristics of diabetic kidney disease (DKD), accumulating data now indicate that tubular atrophy also plays a major role. Cathepsin D (CatD) is the major aspartate protease within lysosomes. The current study demonstrated that CatD expression was altered in the renal tubular epithelium in patients with diabetes mellitus (DM). In contrast to its low and uniform distribution in the tubular epithelium in normal kidney tissues, CatD demonstrated flecked and increased expression in tubules with relatively integral structures, and disappeared in disordered tubules in DM kidney tissues. In vitro studies demonstrated that CatD protected HK2 cells from the damage induced by high glucose and advanced glycation end-products (AGEs), independent of its enzymatic activity. In addition, the current study demonstrated that AGEs induced lysosome membrane permeabilization (LMP) and loss of mitochondrial membrane potential (MMP). Overexpression of CatD prevented LMP and maintained the MMP in HK2 cells exposed to AGEs. In addition, the catalytic activity of CatD was not required for its role in LMP prevention and MMP maintenance. These results indicate, for the first time that CatD may improve the viability of renal tubular cells in the presence of diabetic mediators independent of its enzymatic activity by preventing LMP and stabilizing the MMP.

Cordyceps sinensis attenuates renal fibrosis and suppresses BAG3 induction in obstructed rat kidney.

BAG3 regulates a number of cellular processes, including cell proliferation, apoptosis, adhesion and migration, and epithelial-mesenchymal transition (EMT). However, the role of BAG3 in renal tubular EMT and renal interstitial fibrosis remains elusive. This study aimed to examine the dynamic expression of BAG3 during renal fibrosis, and to investigate the efficacy of Cordyceps sinensis (C. sinensis) on renal fibrosis. A rat model of unilateral ureteral obstruction (UUO) was established, and the expression of BAG3 and α-SMA, and the efficacy of C. sinensis on renal fibrosis induced by UUO were examined. The results showed that UUO led to collagen accumulation, which was significantly suppressed by C. sinensis. UUO increased the expression of BAG3 and α-SMA, a mesenchymal marker, while UUO induced BAG3 and α-SMA expression was significantly inhibited by C. sinensis. In addition, immunohistochemical staining demonstrated that BAG3 immunoreactivity was restricted to tubular epithelium. In conclusion, BAG3 is a potential target for the prevention and/or treatment of renal fibrosis, and C. Sinensis is a promising agent for renal fibrosis.

Implication of Bcl-2-associated athanogene 3 in fibroblast growth factor-2-mediated epithelial-mesenchymal transition in renal epithelial cells.

The epithelial-mesenchymal transition (EMT) of tubular epithelial cells to myofibroblast-like cells plays a substantial role in renal tubulointerstitial fibrosis, which is a common pathological character of end-stage renal disease (ESRD). Fibroblast growth factor-2 (FGF-2) triggers EMT in tubular epithelial cells and increases Bcl-2-associated athanogene 3 (BAG3) expression in neural progenitor and neuroblastoma cells. In addition, a novel role of regulation of EMT has been ascribed to BAG3 recently. These previous reports urged us to study the potential involvement of BAG3 in EMT triggered by FGF-2 in renal tubular epithelial cells. The current study found that FGF-2 induced EMT, simultaneously increased BAG3 expression in human kidney 2 (HK2) cells. Although FGF-2 induced EMT in nontransfected or scramble short hairpin RNA (shRNA) transfected HK2 cells, it was ineffective in BAG3-silenced cells, indicating a favorable role of BAG3 in EMT of tubular cells induced by FGF-2. Knockdown of BAG3 also significantly suppressed motion and invasion of HK2 cells mediated by FGF-2. Furthermore, we confirmed that BAG3 was upregulated in kidney of unilateral ureteral obstruction (UUO) rats, a well-established renal fibrosis model, in which EMT is supposed to exert a substantial influence on renal fibrosis. Importantly, upregulation of BAG3 was limited to tubular epithelial cells. Results of the current study identify BAG3 as a potential player in EMT of tubular epithelial cells, as well as renal fibrosis.

BAG3 regulates ECM accumulation in renal proximal tubular cells induced by TGF-ß1.

Previously we have demonstrated that Bcl-2-associated athanogene 3 (BAG3) is increased in renal fibrosis using a rat unilateral ureteral obstruction model. The current study investigated the role of BAG3 in renal fibrosis using transforming growth factor (TGF)-ß1-treated human proximal tubular epithelial (HK-2) cells. An upregulation of BAG3 in vitro models was observed, which correlated with the increased synthesis of extracellular matrix (ECM) proteins and expression of tissue-type plasminogen activator inhibitor (PAI)-1. Blockade of BAG3 induction by shorting hairpin RNA suppressed the expression of ECM proteins but had no effect on PAI-1 expression induced by TGF-ß1. Forced overexpression of BAG3 selectively increased collagens. TGF-ß1-induced BAG3 expression in HK-2 cells was attenuated by ERK1/2 and JNK MAPK inhibitors. In addition, forced BAG3 overexpression blocked attenuation of collagens expression by ERK1/2 and JNK inhibitors. These data suggest that ERK1/2 and JNK signaling events are involved in modulating the expression of BAG3, which would ultimately contribute to renal fibrosis by enhancing the synthesis and deposition of ECM proteins.

Genetic variants in interleukin genes and susceptibility to IgA nephropathy: a meta-analysis.

Many existing studies have demonstrated that genetic variants in interleukin (IL) genes might have an impact on an individual's susceptibility to IgA nephropathy (IgAN); but individually published results are inconclusive. This meta-analysis aimed to derive a more precise estimation of the relationships between IL genetic variants and IgAN risk. We searched CISCOM, CINAHL, Web of Science, PubMed, Google Scholar, EBSCO, Cochrane Library, and China BioMedicine (CBM) and China National Knowledge Infrastructure (CNKI) databases from inception through August 1, 2013. Meta-analysis was performed using the STATA 12.0 software. Seven case-control studies were included with a total of 1135 IgAN patients and 1603 healthy controls. Our meta-analysis results revealed that genetic variants in IL-1 and IL-1RN genes were associated with an increased risk of IgAN. However, similar associations were not observed in IL-6, IL-10, and IL-22R genes. Subgroup analysis by ethnicity suggested that there were significant associations between IL genetic variants and an increased risk of IgAN among both Asian and Caucasian populations. Meta-regression analyses showed that gene types may be a major source of heterogeneity. No publication bias was detected in this meta-analysis. The present meta-analysis suggests that IL genetic variants may contribute to the risk of IgAN, especially in IL-1 and IL-1RN genes.

The role of CD44-hyaluronic acid interaction in exogenous mesenchymal stem cells homing to rat remnant kidney.

BACKGROUND/AIMS: The aim of our study was to reveal the role of CD44-Hyaluronic acid (HA) in the homing and improving renal function of systemically transplanted MSCs in chronic renal failure. METHODS: First, a remnant kidney model was established in rats and the expression of HA was determined using immunohistochemistry (IHC) and western blotting. Next, chemotaxis assay using flow cytometry, and cell migration assay of MSCs were performed in vitro. Then, MSCs were transplanted into rats, thus, sprague-Dawley (SD) rats were randomly divided into sham group, 5/6 nephrectomy (5/6 Nx) group, MSC group and MSC/Anti-CD44 group (n = 8 for all groups). Migration of MSCs to the kidney in these rats was assessed by using cell tracking experiments, and tissue damage was evaluated by morphological analysis using Masson's trichrome staining and periodic acid Schiff staining. RESULTS: HA was significantly observed in 5/6 Nx group, but not in sham group. Meanwhile, HA was discovered induced MSCs migration remarkably (p < 0.05) and anti-CD44 antibody inhibited the migration significantly (p < 0.05) in vitro. In vivo, the GFP-MSCs were observed in MSC group and the cells reduced in MSC/Anti-CD44 groups, especially, in the tubulointerstitium. CONCLUSION: Our findings reveal that CD44-HA has the potential to induce MSCs homing to injured tissue, while its effect on the ability of MSCs, improving tissue function, is not significant.

[A multi-center, randomized, controlled, double blind and double dummy clinical trial of antofloxacin hydrochloride tablet versus levofloxacin tablet for the treatment of acute bacterial infections].

OBJECTIVE: To evaluate the clinical efficacy and safety of antofloxacin hydrochloride tablet for the treatment of acute bacterial infections. METHODS: A multi-center randomized control, double blind and double dummy clinical trial was conducted; levofloxacin tablet was closed as controlled drug. The duration of treatment was 7-14 days in both groups. RESULTS: A total of 719 patients were enrolled in the study, in which 359 patients treated with antofloxacin and 360 patients treated with levofloxacin were included. Three hundred and thirty and 337 patients completed the study and met with all the criteria for per-protocol analysis, respectively. By the end of chemotherapy, the cured rates in per protocol set (PPS) population were 79.7% and 77.4%, the effective rates were 95.2% and 96.7%, and the bacterial clearance were 96.7% and 97.5% for the treating and control group, respectively. The clinical and bacterial efficacy of antofloxacin and levofloxacin was comparable by the analysis of infectious sites. Three hundred and fifty-seven and 356 patients in antofloxacin and levofloxacin groups were evaluated the safety. The drug adverse events occurred both in 10.1%, and drug adverse reactions occurred in 7.8% and 7.9% patients in the two groups. The most common drug adverse reactions were mild gastroenteric symptoms. No QTc prolongation was detected in all the patients. One patient in each group had mild blood glucose increase at the end of therapy, but the glucose returned to normal level without any intervention. No statistic significant difference between the two groups in clinical efficacy and safety was detected (P > 0.05). CONCLUSIONS: Antofloxacin hydrochloride tablet was effective and safe for the treatment of acute bacterial infections.

[Value of urinary kidney injury molecule-1 protein in early diagnosis of radiocontrast- induced nephropathy in rats].

OBJECTIVE: To assess the value of determination of urine kidney injury molecule-1 (KIM-1) protein content in the early diagnosis of radiocontrast-induced nephropathy (RCIN) in rats. METHODS: Seventy-two adult male SD rats were randomly divided into groups A, B, C (n=8) and D group (which was subdivided into 2, 6, 12, 24, 48 h and 7 days groups, n=8). Group A was subject to injections via the tail vein of PBS and normal saline (NS), group B received injections of PBS, NS, and contrast medium (CM), group C with indomethacin (INDO), nitric oxide synthase inhibitor (L-NAME), and NS, and group D with INDO, L-NAME and CM. Each injection was given at the interval of 15 min. RESULTS: In group D, serum creatinine (Scr) and urea nitrogen (BUN) were significantly increased at 6 h after the injections (P<0.001), peaked at 24 h and recovered normal levels at 48 h. Histological examination revealed significant pathological changes in the kidneys at 6 h, showing diffuse tubulointerstitial hyperemia and hemorrhage, marked tubular necrosis and tubular structure destruction; at 12 h, significant tubular necrosis was still present with tubular structure destruction, but the tubulointerstitial hemorrhage was alleviated; at 24 h, tubular regeneration occurred in the renal medulla, but even till 7 days the tubular structures failed to show full recovery. In group D, urine KIM-1 level began to increase at 2 h, reached the peak level at 24 h, and lasted till 7 days (P<0.001); urine N-acetly-ß-D-glucosaminidase (NAG) value began to increase at 6 h and became normal at 48 h. Urine MMP-9 level underwent no significant changes in group D over the time points of observation. CONCLUSION: The results show that urinary KIM-1 levels can be used as an indicator for early diagnosis of RCIN.

Involvements of Rho-kinase and TGF-beta pathways in aldosterone-induced renal injury.

Recent studies have suggested a role for aldosterone in the pathogenesis of renal injury. This study investigated the potential contributions of Rho-kinase and TGF-beta pathways to aldosterone-induced renal injury. Rats were uninephrectomized and then treated for 5 wk with 1% NaCl in a drinking solution and one of the following: Vehicle (2% ethanol, subcutaneously; n = 9); aldosterone (0.75 microg/h, subcutaneously; n = 9); or aldosterone + fasudil, a specific Rho-kinase inhibitor (10 mg/kg per d, subcutaneously; n = 8). Phosphorylation of myosin phosphate target subunit-1 (MYPT1) and Smad2/3 in renal cortical tissue was measured by Western blotting with anti-phospho MYPT1 and Smad2/3 antibodies, respectively. Rats that received aldosterone infusion exhibited hypertension and severe renal injury characterized by proteinuria, glomerular sclerosis, and tubulointerstitial fibrosis with increases in alpha-smooth muscle actin staining and numbers of monocytes/macrophages in the interstitium. Renal cortical mRNA levels of types I and III collagen, TGF-beta, connective tissue growth factor, and monocyte chemoattractant protein-1 as well as Smad2/3 phosphorylation were significantly increased in rats that received aldosterone infusion. All of these changes were associated with an increase in renal tissue MYPT1 phosphorylation. Treatment with fasudil did not alter BP but significantly ameliorated proteinuria and renal injury in rats that received aldosterone infusion. Furthermore, fasudil prevented MYPT1 phosphorylation and markedly decreased alpha-smooth muscle actin staining, numbers of monocytes/macrophages, mRNA levels of types I and III collagen, TGF-beta, connective tissue growth factor and monocyte chemoattractant protein-1, and Smad2/3 activity in renal cortical tissues. These results provide evidence, for the first time, that Rho-kinase is substantially involved in aldosterone-induced renal injury through activation of a TGF-beta-dependent pathway.