Fabrication and Application of Ag@SiO2/Au Core-Shell SERS Composite in Detecting Cu2+ in Water Environment.

A sensitive and simple method for detecting Cu2+ in the water source was proposed by using surface-enhanced Raman scattering spectroscopy (SERS) based on the Ag@SiO2/Au core-shell composite. The Ag@SiO2 SERS tag was synthesized by a simple approach, in which Ag nanoparticles were first embedded with Raman reporter PATP and next coated with a SiO2 shell. The Ag@SiO2 nanoparticles had strong stability even in a high-concentration salty solution, and there were no changes to their properties and appearance within one month. The Ag@SiO2/Au composite was fabricated through a controllable self-assemble process. L-cysteine was decorated on the surface of a functionalized Ag@SiO2/Au composite, as the amino and carboxyl groups of it can form coordinate covalent bond with Cu2+, which shows that the Ag@SiO2/Au composite labelled with L-cysteine has excellent performance for the detection of Cu2+ in aqueous media. In this study, the SERS detection of Cu2+ was carried out using Ag@SiO2 nanoparticles, and the limit of detection (LOD) as low as 0.1 mg/L was achieved.

Effect of different ankle braces on lower extremity kinematics and kinetics following special-induced fatigue for volleyball players with functional ankle instability.

Background: The aim of this study is to examine the effects of different ankle braces on functional ankle instability (FAI) participants following special-induced fatigue, which will provide advice for preventing ankle sprains in volleyball game. Methods: A total of 18 male collegiate volleyball players with FAI were recruited. The kinematics and kinetics data were acquired from the participants during single-leg drop landing using the infrared motion capture system (Mars2H, Nokov, China) and the force platform (Bertec, USA). A 2 × 2 within subjects design ANOVA was adopted to analyze the data. Results: Whether fatigue or not, soft and semi-rigid brace reduced the ankle inversion (P = 0.025). Moreover, soft brace reduced the sagittal range of motion (ROM) of the ankle joint before fatigue (P = 0.05). In addition, the semi-rigid brace shortened the time to stability in the medial and lateral directions (P = 0.039) as well as the vertical directions (P < 0.001). The semi-rigid brace reduced the ground reaction force post-fatigue (P = 0.001). Conclusion: Soft ankle brace reduced the sagittal range of motion pre-fatigue. Since volleyball requires athletes to jumping and landing repeatedly, and the ankle sagittal ROM was an important cushion during landings. Thus, soft ankle brace might result in overuse injury for lower extremity. However, the semi-rigid ankle brace increased the dynamic stability in the medial and vertical directions, and reduced the ankle inversion angle and forward ground reaction force post-fatigue. This ensured that the volleyball player's ankle was in a neutral position during landing, reducing the risk of excessive inversion caused by contact with the opposing player during spike and block.

Preliminary investigation of the immune activity of PmH2A-derived antimicrobial peptides from the pearl oyster Pinctada fucata martensii.

Antimicrobial peptides (AMPs) play important roles in the immune defense against pathogenic microorganisms. For instance, histone 2A (H2A)-derived AMPs is an antimicrobial peptide involved in the host's innate immune defense and immunoregulation. AMPs have been isolated from the pearl oyster Pinctada fucata martensii but their role in host defense remains poorly understood. To elucidate the structural features of P. f. martensii H2A (PmH2A)-derived AMPs and their potential immune functions, we synthesized a series of laboratory-designed synthetic analogs of PmH2A and examined their antimicrobial properties, as well as their mechanisms of action. This analysis revealed inhibitory effects on the growth of Gram-positive and Gram-negative bacteria. Further assessment by transmission electron microscopy (TEM) of two of the three peptides, PmH2A-AMP and PmH2A-AMP(5-13)[KLLK]3, confirmed that it exerted an anti-bacterial activity through membrane lysis. Finally, we found that the hemocytes and gills of P. f. martensii released antimicrobial H2A histones in response to LPS exposure, mimicking tissue damage and infection. This immune response is reminiscent of the neutrophil extracellular traps (NETs) recently described in oysters. Thus, the LPS challenge is sufficient to induce histone-derived peptide accumulation in pearl oyster P.f. martensii.

Members of the histone-derived antimicrobial peptide family from the pearl oyster Pinctada fucata martensii: Inhibition of bacterial growth.

Because it is difficult to isolate standard antimicrobial peptides (AMPs) using traditional biochemical approaches, we designed, synthesized, and evaluated a series of structurally altered histone-derived AMPs (HDAPs) from the pearl oyster Pinctada fucata martensii using molecular cloning approaches. Four histone-homolog genes (PmH2A, PmH2B, PmH3, and PmH4-1) were identified, of which PmH2A and PmH2B had yet to be described. PmH2A and PmH2B were therefore cloned using Rapid Amplification of cDNA Ends (RACE) and characterized. Constitutive PmH2A and PmH2B mRNA expression was detected in all six pearl oyster tissues tested, with comparatively greater transcript abundance in the gonads. Because α-helical content, hydrophilicity index, and the presence of a proline hinge may be the three important factors influencing the antimicrobial efficacy of HDAPs, we synthesized a series of eight N- and C-terminally truncated or amino acid-substituted synthetic candidate HDAP analogs derived from PmH2A, PmH2B, PmH3, and PmH4-1. Only the PmH2A- and PmH4-derived AMPs inhibited bacterial growth. The PmH2A-derived AMPs were α-helical proteins, while the PmH4-derived AMPs were extended strand/random coil proteins. Our results suggested that having an α-helical structure was particularly important for the antibacterial efficacy of the PmH2A-derived peptides; amphipathic structures (hydrophilic index, 0.3 to -0.3) may enhance the antimicrobial function of both the PmH2A- and PmH4-derived peptides. The high antibacterial efficacy of one of the HDAP analogs studied, PmH2A-AMP (5-13) [KLLK]3, indicated that this protein may represent a promising candidate for the treatment of bacterial infections in aquaculture mollusk species. This first study of HDAPs from the pearl oyster P. f. martensii provides new insights into the design and function of highly effective antimicrobial peptides.

Gene cloning and functional study of PmKSPI from Pinctada fucata martensii.

Kunitz-type serine protease inhibitors (KSPI) are a family of serine protease inhibitors (SPIs) and are extensively found in animals, plants, and microbes. SPI can inhibit proteases that may be harmful or unwanted to its cells. Here, a four-domain Kunitz-type SPI, PmKSPI, was cloned by RACE in the pearl oyster Pinctada fucata martensii. The full-length cDNA sequence of PmKSPI was 1318 bp, including the 5' UTR (25 bp), the 3' UTR (96 bp) and ORF (1197 bp). Homology analysis indicated that PmKSPI had the highest resemblance (30.14%) with its homolog in Crassostrea gigas. Phylogenetic analysis revealed that PmKSPI clustered with homologs in other molluscs. We found that PmKSPI mRNA expression in P. f. martensii was distributed in all six tissues, with the highest level in the mantle, and almost no expression in other tissues. After PAMPs challenge, expression of PmKSPI mRNA in the mantle was significantly up-regulated. The recombinant protein rPmKSPI significantly inhibited the growth of 5 kinds of Gram-negative bacteria but had little effect on Gram-positive bacterial activity. Transmission electron microscopy showed that plasmolysis occurred in two Gram-negative bacteria species when treated with rPmKSPI. rPmKSPI may thus have a bactericidal effect by destroying the bacterial cell membrane or cell walls and releasing its contents. Therefore, our results suggest that PmKSPI is tightly associated with the immunological defence of P. f. martensii.

Review of Thin-Layer Chromatography Tandem with Surface-Enhanced Raman Spectroscopy for Detection of Analytes in Mixture Samples.

In the real world, analytes usually exist in complex systems, and this makes direct detection by surface-enhanced Raman spectroscopy (SERS) difficult. Thin layer chromatography tandem with SERS (TLC-SERS) has many advantages in analysis such as separation effect, instant speed, simple process, and low cost. Therefore, the TLC-SERS has great potential for detecting analytes in mixtures without sample pretreatment. The review demonstrates TLC-SERS applications in diverse analytical relevant topics such as environmental pollutants, illegal additives, pesticide residues, toxic ingredients, biological molecules, and chemical substances. Important properties such as stationary phase, separation efficiency, and sensitivity are discussed. In addition, future perspectives for improving the efficiency of TLC-SERS in real sample detecting are outlined.

Comprehensive analysis of the lncRNAs, mRNAs, and miRNAs implicated in the immune response of Pinctada fucata martensii to Vibrio parahaemolyticus.

Non-coding RNAs (ncRNAs) have been implicated in a variety of biological processes. However, most ncRNAs are of unknown function and are as-yet unannotated. The immune-related functions of ncRNAs in the pearl oyster Pinctada fucata martensii were explored based on transcriptomic differences in the expression levels of long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) in the hemocytes of P.f. martensii after challenge by the pathogenic bacterium Vibrio parahaemolyticus. Across the challenged and control pearl oysters, 144 miRNAs and 14,571 lncRNAs were identified. In total, 13,375 ncRNAs were differentially expressed between the challenged and control pearl oysters; in the challenged pearl oysters as compared to the controls, 15 miRNAs and 5147 lncRNAs were upregulated, while 51 miRNAs and 8162 lncRNAs were downregulated. The sequencing results were validated using quantitative real-time polymerase chain reaction (qRT-PCR) analysis. GO and KEGG pathway analysis showed that genes targeted by the differentially expressed ncRNAs were associated with the vascular endothelial growth factor (VEGF) signaling pathway and the nuclear factor kappa-B (NF-κB) signaling pathway. An lncRNA-mRNA-miRNA network that was developed based on the transcriptomic results of this study suggested that lncRNAs may compete with miRNAs for mRNA binding sites. This study may provide a useful framework for the detection of additional novel ncRNAs, as well as new insights into the pathogenic mechanisms underlying the response of P.f. martensii to V. parahaemolyticus.

Layered mechanical and electrical properties of porcine articular cartilage.

The complex structure and composition of articular cartilage make its performance show depth-dependent characteristics, but its related parameters are not complete at present. In this study, porcine articular cartilage was divided into three zones along the thickness direction, and the cartilage tissue in each zone was tested for electrical impedance, compression relaxation, and permeability to obtain their mechanical and electrical impedance parameters. The results showed that there were significant differences in mechanical and electrical properties of cartilage tissue in different zones in which resistivity, elastic modulus, relaxation time, and final relaxation rate increased gradually from superficial zone to deep zone along the direction of cartilage thickness while the permeability decreased gradually. Bioimpedance analysis can capture the phenomenon of very slight histological changes, which is expected to provide information for predicting cartilage degeneration, but the electrical impedance parameters of cartilage are still very lacking. These data are expected to provide reference for the treatment of clinical osteoarthritis and the research of cartilage repair materials.

The succinylome of Pinctada fucata martensii implicates lysine succinylation in the allograft-induced stress response.

Lysine succinylation is a novel protein post-translational modification associated with the regulation of a variety of cellular processes. Post-translational modifications may regulate the immune response of Pinctada fucata martensii, a marine bivalve used to produce cultured pearls, in response to the surgical implantation of the seed pearl. This allograft-induced stress response may lead to transplant rejection or host death. However, the regulatory effects of post-translational modifications following nucleus insertion surgery in P.f. martensii remain largely unknown. Here, we used 4D label-free quantitative proteomics (4D-LFQ) with LC-MS/MS to explore the effects of nucleus implantation on lysine succinylation in P.f. martensii. We identified 4430 succinylated sites on 964 succinylated proteins in P.f. martensii after nucleus insertion surgery, and seven conserved motifs were identified upstream and downstream of these sites. In total, 269 succinylation sites were differentially expressed in response to implantation (|fold-change| > 1.5 and FDR <1%; 211 upregulation and 58 downregulation), corresponding to 163 differentially expressed succinylated proteins (DESPs; 124 upregulated and 39 downregulated). The terms over-enriched in the DESPs included "cellular processes", "metabolic pathways", and "binding activity", while the significantly enriched pathways included "ECM-receptor interaction", "PI3K-Akt signaling", and "focal adhesion". "EGF-like structural domains", "platelet-responsive protein type 1 structural domains", and "laminin EGF-like (domains III and V) domains" were overrepresented in the DESPs. Parallel reaction-monitoring (PRM) analysis validated 13 DESPs from the proteomics data. The succinylome of P.f. martensii (generated here for the first time) helps to clarify the biological role of large-scale succinylation in this bivalve after nucleus insertion surgery, providing a theoretical basis for further investigations of stress-induced post-translational modifications in other mollusks and extending our knowledge of the molluscan succinylated proteome.

Cloning and functional analysis of a trypsin-like serine protease from Pinctada fucata martensii.

Trypsin-like serine proteases (TLSs) play various roles in dietary protein digestion, hemolymph coagulation, antimicrobial peptide synthesis, and, in particular, the rapid immune pathways activated in response to pathogen detection. The cultured pearl industry, of which Pinctada fucata martensii is one of the most important species, is plagued by disease, thus leading to large economic losses. Herein, the molecular mechanisms underlying the innate immune response of P.f. martensii were explored. First, immune effector molecules from the P.f. martensii genome were screened and a TLS-like gene encoding a protein with a trypsin domain, herein designated as PmTLS, was identified. A multi-sequence alignment indicated a low sequence homology between PmTLS and other mollusk TLS-like proteins. Furthermore, a neighbor-joining phylogenetic analysis indicated that PmTLS has the closest genetic relationship to a Crassostrea gigas TLS. Additionally, real-time quantitative PCR (qPCR) analysis showed that PmTLS mRNA is constitutively expressed in all of the 6 examined P.f. martensii tissues, with significantly higher expression noted in hemocytes relative to the other tissues examined (p < 0.05). P.f. martensii samples were then challenged with various pathogen-associated molecular patterns (PAMPs), including lipopolysaccharide, peptidoglycan, and polyinosinic acid. In the challenge groups, PmTLS was significantly upregulated in hemocytes at 48 h post-challenge when compared to the unchallenged controls. Furthermore, treatment with recombinant PmTLS (rPmTLS) also significantly inhibited the growth of most of the examined gram-negative bacteria tested in vitro (p < 0.05), but it had little effect on the growth of the examined gram-positive bacteria. When examining morphological changes via transmission electron microscopy, rPmTLS treated bacteria exhibited morphological changes such as plasma wall separation. Thus, rPmTLS appears to play a bactericidal role by destroying bacterial cell membranes or cell walls, which subsequently leads to a release of the cellular contents and cell death. The findings presented herein have enabled further characterization of the immune defense mechanisms in P.f. martensii and may lead to improved disease control methods for the pearl cultivation industry.

Ultra-Sensitive, Rapid and On-Site Sensing Harmful Ingredients Used in Aquaculture with Magnetic Fluid SERS.

The integration of surface-enhanced Raman scattering (SERS) spectroscopy with magnetic fluid provides significant utility in point-of-care (POC) testing applications. Bifunctional magnetic-plasmonic composites have been widely employed as SERS substrates. In this study, a simple and cost-effective approach was developed to synthesize magnetic-plasmonic SERS substrates by decorating silver nanoparticles onto magnetic Fe3O4 nanoparticles (AgMNPs), which function both as SERS-active substrates and magnetic fluid particles. The strong magnetic responsivity from AgMNPs can isolate, concentrate, and detect target analytes from the irregular surface of fish skin rapidly. We fabricate a microfluid chip with three sample reservoirs that confine AgMNPs into ever smaller volumes under an applied magnetic field, which enhances the SERS signal and improves the detection limit by two orders of magnitude. The magnetic fluid POC sensor successfully detected malachite green from fish with excellent selectivity and high sensitivity down to the picomolar level. This work achieves a label-free, non-destructive optical sensing approach with promising potential for the detection of various harmful ingredients in food or the environment.

Comparative proteomics and transcriptomics illustrate the allograft-induced stress response in the pearl oyster (Pinctada fucata martensii).

Implantation of a spherical nucleus into a recipient oyster is a critical step in artificial pearl production. However, the molecular mechanisms underlying the response of the pearl oyster to this operation are poorly understood. In this research, we used transcriptomic and proteomic analyses to examine allograft-induced changes in gene/protein expression patterns in Pinctada fucata martensii 12 h after nucleus implantation. Transcriptome analysis identified 688 differential expression genes (DEGs) (FDR<0.01 and |fold change) ＞ 2). Using a 1.2-fold increase or decrease in protein expression as a benchmark for differentially expressed proteins (DEPs), 108 DEPs were reliably quantified, including 71 up-regulated proteins (DUPs) and 37 down-regulated proteins (DDPs). Further analysis revealed that the GO terms, including "cellular process", "biological regulation" and "metabolic process" were considerably enriched. In addition, the transcriptomics analysis showed that "Neuroactive ligand-receptor interaction", "NF-kappa B signaling pathway", "MAPK signaling pathway", "PI3K-Akt signaling pathway', "Toll-like receptor signaling pathway", and "Notch signaling pathway" were significantly enriched in DEGs. The proteomics analysis showed that "ECM-receptor interaction", "Human papillomavirus infection", and "PI3K-Akt signaling pathway" were significantly enriched in DEPs. The results indicate that these functions could play an important role in response to pear oyster stress at nucleus implantation. To assess the potential relevance of quantitative information between mRNA and proteins, using Ward's hierarchical clustering analysis clustered the protein/gene expression patterns across the experimental and control samples into six groups. To investigate the biological processes associated with the protein in each cluster, we identified the significantly enriched GO terms and KEGG pathways in the proteins in each cluster. Gene set enrichment analysis (GSEA) was used to reveal the potential protein or transcription pathways associated with the response to nuclear implantation. Thus, the study of P. f. martensii is essential to enhance our understanding of the molecular mechanisms involved in pearl biosynthesis and the biology of bivalve molluscs.

Study on the poroelastic behaviors of the defected articular cartilage.

This article presented the possible mechanism of arthritis damaged changes in cartilage's interstitial fluid flowing behavior. Firstly, the analytical solutions for the pore fluid pressure and velocity in the idealized cartilage defect model were obtained, which are employed to validate the finite element (FE) method. Then according to the MRI data, an articular cartilage FE model was developed to study the effects of defect characteristics on its poroelastic behaviors. The results showed the interstitial fluid pressure and velocity in defected articular cartilage is diminished, moreover, this trend is even more severe as the defect radius or thickness increased. As the development of osteoarthritis goes, the fluid velocity is decreased and cause the even serious nutrients loss.

A randomized controlled trial on the effect of dietary guidance on the treatment of Henoch-Schonlein purpura in children.

The amino acid-based formulae were extensively added to diet of children for the treatment of Henoch-Schonlein purpura (HSP), and the nutrition and growth situation of children were evaluated after giving new dietary intervention. Patients were randomly divided into restricted diet group (n=30) and dietary guidance group (n=30). Besides, 30 cases with bronchiolitis who had normal diet were selected as the control group. The dietary questionnaire was designed to record the types and intakes of various foods taken by children every day, and the intake levels of nutrients were analyzed. Physical examination, biochemical analysis of blood and urine routine were carried out to evaluate the effect of dietary guidance on their growth and development. The results showed that restricted diet group had lower levels of nutrient intake and the actual/recommended percentage. However, overall nutrient intake level of the dietary guidance group was higher, basically equal to the recommended intake level. Besides, the actual intake and actual/recommended percentage of nutrients of dietary guidance group were significantly higher than those of restricted diet group (p<0.05). Dietary guidance can improve nutrients and protein intake of children with HSP, and reduce the relapse of rash and incidence of complications.

Efficacy and Safety of TangWang Prescription for Type 2 Non-Proliferative Diabetic Retinopathy: A Study Protocol for a Randomized Controlled Trial.

Background: Diabetic retinopathy (DR) is one of the most common and severe microvascular complications of diabetes mellitus (DM), which results in blindness among adults worldwide. Presently, the efficacy of drug treatments for diabetic retinopathy (DR) is not satisfactory, thus urgently necessitating effective drug treatment measures. TangWang prescription (TWP) has been found to have retinal protection effects in previous clinical and basic research. However, there is a lack of rigorous, randomized, and controlled studies. This study aims to evaluate the efficacy and safety of TWP in delaying the development of DR. Methods: This study is a randomized, double-blind, placebo-controlled, parallel-group, multicenter clinical trial, consisting of 384 participants to be randomized in a 1:1 ratio in the treatment and control groups. Furthermore, the treatment and control groups will be administered the TangWang prescription and the placebo, respectively, each at a dose of one bag twice a day. The study period will last for 48 weeks. The primary outcome measure will be the changes in the degree of retinal microvascular lesions before and after treatment. The secondary outcome will be changes in the degree of hemangioma, microvascular bleeding, microvascular leakage, macular edema, and vision. All statistical tests will be two-sided, and a p < 0.05 will be considered statistically significant. Discussion: We hypothesize that the patients with DR will benefit from TangWang prescription, and in addition to the central random system and platform of dynamic information collection, the patients' conditions will be monitored, and the data collected for analysis. If successful, this study will provide evidence that the TWP formulation delays in the progression of DR. Trial registration: The design of this trial has been registered with the ClinicalTrials.gov (NCT03025399).

Copeptin and Nesfatin-1 Are Interrelated Biomarkers with Roles in the Pathogenesis of Insulin Resistance in Chinese Children with Obesity.

BACKGROUND: Copeptin and nesfatin-1 have recently been identified as novel peptides that play a role in the pathogenesis of obesity-related insulin resistance in adults. However, the relationship between them has not yet been elucidated, and their circulating levels in children with obesity have not been adequately studied. Therefore, the current study aimed to investigate whether their levels are altered in Chinese children with obesity, as well as to determine the correlation of these 2 peptides with each other, with insulin resistance, and with other biochemical parameters. METHODS: A total of 156 children were enrolled in this study, including 101 children with obesity and 55 lean controls. Anthropometric parameters and clinical data of all subjects were collected, and circulating tumor necrosis factor-α, adiponectin, leptin, copeptin, and nesfatin-1 levels were measured using ELISA. RESULTS: Serum copeptin and nesfatin-1 levels were significantly elevated in children with obesity and children with insulin resistance compared to control subjects. In addition, nesfatin-1 and copeptin levels were found to be significantly positively correlated with one another by Pearson's correlation and partial correlation. In multiple regression analysis using nesfatin-1 or copeptin as the dependent parameter, a significant correlation was observed between nesfatin-1 and copeptin, and associations between each of them with homeostasis model assessment of insulin resistance (HOMA-IR) were detected. CONCLUSION: These novel findings shed light on the possible interplay role of these 2 molecules in obesity-related insulin resistance.

Moxibustion for the treatment of diabetic peripheral neuropathy: A systematic review and meta-analysis following PRISMA guidelines.

BACKGROUND: At present, the effect of western-medicine (WM) therapy to treat diabetic peripheral neuropathy (DPN) is limited. Moxibustion is a representative external treatment in traditional Chinese medicine that has been beneficial to DPN. We aim to systematically assess the efficacy and safety of moxibustion in treating DPN, following PRISMA guidelines. METHODS: Eight electronic databases were searched to acquire information on eligible trials published from inception to June 1, 2019. We included randomized controlled trials (RCTs) applying moxibustion therapy with a minimum of 14-days treatment duration for DPN patients compared with placebo, no intervention, or conventional WM interventions. The primary outcomes in our study include the sensory-nerve conduction velocity (SNCV) and motor-nerve conduction velocity (MNCV). We used the Cochrane Collaboration Risk of Bias tool to assess the methodological quality of eligible RCTs. Statistical analyses were conducted using Review Manager 5.3. Risk ratios (RR) and mean differences (MD) were calculated with a 95% confidence interval (CI). The χ test was applied to assess the heterogeneity. RESULTS: In total, 11 RCTs were included that involved 927 DPN patients. Compared with the control group, there was an increase in median MNCV (MD = 6.26, 95% CI 2.64-9.89, Z = 3.39, P = .0007) and peroneal MNCV (MD = 6.45, 95% CI 5.30-7.61, P < .00001). There was also an increase in median SNCV (MD = 6.64, 95% CI 3.25-10.03, P = .0001) and peroneal SNCV (MD = 3. 57, 95% CI 2.06-5.09, Z = 4.63, P < .00001) in the treatment groups. The treatment groups receiving moxibustion therapy indicated a more significant improvement in total effectiveness rate (RR = 0.25, 95% CI 0.18-0.37, Z = 7.16, P < .00001). Toronto Clinical Scoring System indicated a significant decrease in the treatment groups (MD = -2.12, 95% CI -2.82 to 1.43, P < .00001). Only 1 study reported that treatment groups experienced no adverse reactions. The other 10 studies did not mention adverse events. CONCLUSIONS: Moxibustion therapy may be an effective and safe option for DPN patients but needs to be verified in further rigorous studies.

Banxia Xiexin Decoction Ameliorates t-BHP-Induced Apoptosis in Pancreatic Beta Cells by Activating the PI3K/AKT/FOXO1 Signaling Pathway.

BACKGROUND: Banxia Xiexin Decoction (BXXD) reportedly regulates glycolipid metabolism and inhibits pancreatic ß-cell apoptosis. This study is aimed at investigating the protective effect of BXXD on tert-butyl hydroperoxide- (t-BHP-) induced apoptosis in MIN6 cells and the underlying mechanisms. METHODS: MIN6 cells were preincubated with BXXD or liraglutide (Li) with or without PI3K inhibitor LY294002 (LY) for 12 h, following which t-BHP was added to induce MIN6 cell apoptosis. The protective effects of BXXD on MIN6 cells were evaluated by detecting cell viability and proliferation and glucose-stimulated insulin secretion (GSIS). The antiapoptotic effects were evaluated by Hoechst 33342 staining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay (TUNEL). Malondialdehyde and glutathione peroxidase content and superoxide dismutase activity were measured using commercial kits. The expression of PI3K/AKT/FOXO1 signaling pathway-related signal molecules, and that of apoptotic indicators Bax, P27, and Caspase-3, was quantified using western blotting. RESULTS: Preincubation with BXXD significantly improved t-BHP-induced proliferation inhibition and apoptosis and enhanced GSIS. t-BHP induced the generation of reactive oxygen species and inhibited the activities of antioxidant enzymes, which could be neutralized by pretreatment with BXXD. BXXD promoted the phosphorylation of AKT and FOXO1 in t-BHP-induced MIN6 cells. Moreover, BXXD attenuated the expression of related apoptotic indicators Bax, P27, and Caspase-3. LY abolished these effects of BXXD. CONCLUSION: BXXD protected MIN6 cells against t-BHP-induced apoptosis and improved insulin secretory function through modulation of the PI3K/AKT pathway and the downstream FOXO1, thus suggesting a novel therapeutic approach for type 2 diabetes mellitus (T2DM).

Effectiveness of clean development policies on coal-fired power generation: an empirical study in China.

Coal-fired power will remain China's primary source of electricity for a long time to come. The clean development of coal-fired power generation has become an important strategic choice for China's energy transformation. Based on collecting and sorting out the driving policies for China's clean development of coal-fired power generation from 1997 to 2016, firstly, we reviewed the historical evolution of these policies in terms of the policy quantity, policy theme, and policy instrument. Then, we designed the scoring criteria to quantify the policies. Finally, the effectiveness of the overall policies and different policy instruments was empirically analyzed by the econometric model built on the basis of Cobb-Douglas production function. The results show that the annual release amount of driving policies for the clean development of coal-fired power in China presents a trend of fluctuating growth. Policy themes are mainly divided into two parts, namely production capacity management and air pollutants control; policy instruments mainly include elimination of backward units, project approval, standards, generation price regulation, tax incentives, loan policy, and information publicity; the overall policies have significant positive effects on the clean development of coal-fired power industry; in terms of policy instruments, the elimination of backward units and generation price regulation have significant effects on improving the energy efficiency; project approval and generation price regulation are effective for reducing SO2 emissions rate; and the effects of standards, tax incentives, loan policy, and information publicity on the clean development of coal-fired power are not significant. On these bases, some policy implications to further promote the clean development of coal-fired power industry are provided.

Chinese herbal medicine Tangshen Formula treatment for type 2 diabetic kidney disease in the early stage: study protocol for a randomized controlled trial.

BACKGROUND: Diabetic kidney disease (DKD) is the main cause of end-stage kidney disease and has become a heavy economic and social burden due to its high prevalence and morbidity. The most effective strategy is that patients with DKD should be diagnosed and treated early. Preliminary studies showed that the Chinese herbal Tangshen Formula (TSF) may delay the progression of DKD, reducing microalbuminuria and macroalbuminuria and improving renal function. We designed a randomized, double-blind, placebo-controlled trial to evaluate the efficacy of TSF in patients with DKD. METHODS/DESIGN: This trial is a 13-center, randomized, double-blind, placebo-controlled study. A total of 632 participants will be randomized in a 1:1 ratio to an experiment group (TSF plus losartan) and a control group (placebo plus losartan). The trial cycle will last 24 weeks. The primary outcome will be the change in the urine microalbumin-creatinine ratio from baseline to week 24. The secondary outcome will be the change in the rate of progression to the clinical proteinuria period after intervention, the rate of urine microalbumin negative conversion, the rate of normal urinary microalbumin, the doubling rate of the baseline creatinine value and the glomerular filtration rate between the two groups. Safety in medication will also be evaluated. DISCUSSION: We hypothesize that patients with type 2 diabetes in the early stage of DKD will benefit from TSF. If successful, this study will provide evidence-based recommendations for clinicians. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03009864. Registered January 2017.