Association of Interleukin-1 Beta and Interleukin-1 Receptor Antagonist Gene Polymorphisms and Plasma Levels with Diabetic Nephropathy.

Objective: We investigated the relationships between interleukin- (IL-) 1ß and IL-1 receptor antagonist (IL-1Ra) gene polymorphism and plasma levels in patients with diabetic nephropathy (DN). Methods: The genotype and allele frequency distribution of IL-1ß and IL-1Ra in 61 patients with DN and 48 healthy controls (HCs) were determined by kompetitive allele-specific PCR (KASP), and the plasma concentrations of IL-1ß and IL-1Ra in DN patients and HCs were measured by enzyme-linked immunosorbent assays (ELISA). Results: Significant differences were detected in the distribution of IL-1ß (-511C/T) genotype and allele frequencies between the DN and HC groups (P < 0.05), with the T genotype being more frequent in DN patients than HCs (OR = 2.84, 95% CI: 1.489-5.416). The IL-1ß (+3953C/T) and IL-1Ra (+8006C/T) genotypes and allele frequencies were not significantly different between the two groups (P > 0.05). The plasma IL-1ß level was significantly higher (P < 0.01), while the plasma IL-1Ra concentration was significantly lower in the DN group than the HC group (P < 0.05). Furthermore, the plasma IL-1ß level was significantly different between IL-1ß (-511C/T) locus variants (P < 0.05). Conclusion: The IL-1ß (-511C/T) gene polymorphism was significantly associated with DN risk in the population of northern Guangxi, China, and the T allele maybe responsible for genetic susceptibility to DN.

Diagnostic value of serum cystatin C for diabetic nephropathy: a meta-analysis.

BACKGROUND: Although dozens of studies have investigated the relationship between the content of serum cystatin C (Cys-C) and diabetic nephropathy (DN), the results are still controversial. Hence, This study aims to explore the accuracy of serum Cys-C for diagnosing DN by meta-analysis. METHODS: The studies about serum Cys-C diagnosing DN were searched from six online databases from inception to September 22, 2020. The data were processed by Stata 15.0 statistic software. The corresponding diagnostic effect sizes, such as sensitivity and specificity, were obtained. We drew a summary receiver operating characteristic (SROC) curve. We assess the risk of literature bias was following the QUADAS-2 guidelines. RESULTS: Twenty-six published studies were identified. The results showed a pooled sensitivity of 0.86 (95% confidence interval (CI): 0.82-0.90), specificity of 0.89 (95%CI: 0.85-0.92), positive likelihood ratio of 7.59 (95%CI: 5.66-10.19), negative likelihood ratio of 0.16 (95%CI: 0.12-0.21), and diagnostic odds ratio of 48.03 (95%CI: 30.64-75.29). The area under the SROC curve was given a value of 0.94 (95%CI: 0.91-0.96). CONCLUSION: Serum cystatin C has an excellent diagnostic value with good sensitivity and specificity for diabetic nephropathy.

The effects of omega-3 fatty acids in type 2 diabetes: A systematic review and meta-analysis.

BACKGROUND: The effect of omega-3 polyunsaturated fatty acids (n-3 PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on cardiovascular risk modification in type 2 diabetes and related complications remain unclear. We aim to assess the published effects of n-3 PUFA interventions on lipid risk factors in type 2 diabetes. METHODS: We searched the literature on Pubmed, Embase, CENTRAL, and Web of Science databases in order to perform a pooled analysis of randomized clinical trials (RCTs) assessing n-3 PUFA interventions in type 2 diabetes. The primary outcomes analyzed were the effect of n -3 PUFAs on metabolic biomarkers in type 2 diabetes. RESULTS: 46 RCTs involving 4991 patients with type 2 diabetes were identified for further analysis. Analysis of results showed that n-3 PUFAs interventions significantly improved total cholesterol (TC, WMD = -0.22; 95% CI: -0.32∼ -0.11), triglyceride (TG,WMD = -0.36; 95% CI: -0.48∼-0.25), high-density lipoprotein cholesterol (HDL-C,WMD = 0.05; 95% CI: 0.02∼ 0.08), hemoglobin A1c (HbA1c, WMD = -0.19; 95% CI: -0.31∼-0.06) and C-reactive protein (CRP,WMD = -0.40; 95% CI: -0.74∼-0.07) levels compared to controls (p < 0.05). There was no significant effect on renal function, fasting blood sugar (FBS), insulin resistance (HOMA-IR), low-density lipoprotein cholesterol (LDL-C), adiponectin and leptin (p > 0.05). CONCLUSIONS: The results of this systematic review suggest that n-3 PUFAs can improve cardiovascular risk factors in type 2 diabetes.

Reprogramming the immunosuppressive microenvironment of IDH1 wild-type glioblastoma by blocking Wnt signaling between microglia and cancer cells.

The vast majority (>90%) of glioblastoma (GBM) patients belong to the isocitrate dehydrogenase 1 wild type (IDH1WT) group which exhibits a poor prognosis with a median survival of less than 15 months. This study demonstrated numerous immunosuppressive genes as well as ß-catenin gene, pivotal for Wnt/ß-catenin signaling, were upregulated in 206 IDH1WT glioma patients using the Chinese Glioma Genome Atlas (CGGA) database. The increase in microglia with an immunosuppressive phenotype and the overexpression of ß-catenin protein were further verified in IDH1WT GBM patients and IDH1WT GL261 glioma allografts. Subsequently, we found that IDH1WT GL261 cell-derived conditioned medium activated Wnt/ß-catenin signaling in primary microglia and triggered their transition to an immunosuppressive phenotype. Blocking Wnt/ß-catenin signaling not only attenuated microglial polarization to the immunosuppressive subtype but also reactivated immune responses in IDH1WT GBM allografts by simultaneously enhancing cytotoxic CD8+ T cell infiltration and downregulating regulatory T cells. Positron emission tomography imaging demonstrated enhanced proinflammatory activities in IDH1WT GBM allografts after the blockade of Wnt signaling. Finally, gavage administration of a Wnt signaling inhibitor significantly restrained tumor proliferation and improved the survival of model mice bearing IDH1WT GBM allografts. Depletion of CD8+ T cells remarkably abrogated the therapeutic efficacy induced by the Wnt signaling inhibitor. Overall, the present work indicates that the crosstalk between IDH1WT glioma cells and immunosuppressive microglia is important in maintaining the immunosuppressive glioma microenvironment. Blocking Wnt/ß-catenin signaling is a promising complement for IDH1WT GBM treatment by improving the hostile immunosuppressive microenvironment.

Using iron-based phosphate binders in phosphate reduction and anemia improvement in patients receiving dialysis: a meta-analysis of randomized controlled trials.

PURPOSE: A study was conducted to determine whether iron-based phosphate binders (IBPBs) need to be preferred for hyperphosphatemia and anemia management in patients on dialysis. METHODS: For this meta-analysis, we searched PubMed, Embase, and Cochrane Central Register of Controlled Trials for randomized controlled trials that evaluated the efficacy and safety of IBPBs in decreasing phosphate and correcting anemia in dialysis patients. RESULTS: Nineteen trials comprising 4719 participants were included. Compared with placebo, serum phosphate decreased significantly after treatment with ferric citrate (FC), fermagate (one study), and SBR759 (one study). Hemoglobin increased significantly after treatment with FC and sucroferric oxyhydroxide (PA21). In addition, FC and PA21 reduced serum intact parathyroid hormone (iPTH) and increased ferritin and transferrin saturation, but SBR759 did not. Compared with active treatment, the non-inferiority of IBPBs in reducing serum phosphate and iPTH was demonstrated. FC significantly improved serum hemoglobin and iron-related parameters and decreased the use of intravenous iron and erythropoiesis-stimulating agent, whereas PA21 did not increase serum hemoglobin level. The incidences of infection and hospitalization were similar between the two groups, with FC having a higher risk of diarrhea than the placebo and active treatments. CONCLUSION: FC was associated with the control of hyperphosphatemia and the improvement of anemia. However, PA21 did not show superiority for alleviating anemia compared with the active treatment. Other IBPBs, such as fermagate and SBR759, remained poorly understood due to the limited number of studies. Further trials are required to assess the effect of IBPBs on the risk of cardiovascular events and all-cause mortality.

ComMSnDB-An Automatable Strategy to Identify Compounds from MS Data Sets (Identification of Gypenosides as an Example).

Gynostemma pentaphyllum (Thunb.) Makino is a popular functional food and is also used as an important medicinal plant in China. Gypenoside, the main active constituent in G. pentaphyllum (Thunb.) Makino, belongs to dammarane-type triterpenoid saponins. Due to its high molecular weight and high polarity, it is difficult to obtain complete compound information for gypenoside extracts via mass spectrometry experiments. In this study, an automated targeted data postprocessing strategy called Compound MSn Database (ComMSnDB) was designed and established to elucidate compounds in gypenoside extracts based on ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-ESI-Q-TOF-MS/MS). As a result, 18 types of and 199 main saponin constituents, including 47 potential novel compounds, were tentatively identified from different habitats. At the same time, 15 gypenoside standard compounds were used to verify the feasibility of the ComMSnDB strategy. These results demonstrated that ComMSnDB offers practical value for quick, automated, and effective compound identification.

New MS network analysis pattern for the rapid identification of constituents from traditional Chinese medicine prescription Lishukang capsules in vitro and in vivo based on UHPLC/Q-TOF-MS.

Ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) has recently been widely used in qualitative analyses of traditional Chinese medicine (TCM) prescriptions. However, a poor understanding of detected mass spectral data has rendered data processing difficult and time-consuming. Efficient and widespread data analysis methods focused on identifying both phytochemical compounds and metabolites of TCM prescriptions have rarely been described. In this study, a new MS network analysis pattern that uses model drug Lishukang (LSK) capsules to accelerate the data processing of TCM preparations is developed. The MS network analysis pattern integrates intrinsic structural correlations of phytochemical compounds and structural information derived from mass spectrometry to identify the same types of compounds from a raw data stream. As a result, five MS networks of flavones and flavone glycosides, alkaloids, phenolic acids, saponins and benzylester glucosides in LSK are preliminarily established. 278 compounds, including 9 potential novel compounds are identified or tentatively assigned based on MS networks. Furthermore, 57 potential metabolites of LSK are identified in rat plasma, and potential metabolic pathways are investigated under the guidance of MS networks in vitro. The MS network analysis pattern serves as an integral solution for identifying phytochemical compounds and metabolites of TCM prescriptions. The investigations of LSK also provide essential data for its further study.

Matrix Metalloproteinase-9 -1562C/T Gene Polymorphism Is Associated with Diabetic Nephropathy.

To investigate the association between the metalloproteinase-9 (MMP9) -1562C/T polymorphism and diabetic nephropathy (DN) in Han Chinese, the patients with type 2 diabetes were collected and divided into the non-DN (NDN) and DN groups; controls were recruited. Genotype and allele frequencies were assessed using polymerase chain reaction and restriction fragment length polymorphism. Results showed that SBP, DBP, HbA1c, UAER, Cr, BUN, TG, and TC were higher in the DN group compared with the control and NDN groups. SBP, HbA1c, and TC in DN patients with the TT and CT genotypes were lower than in those with CC. Compared with controls, the frequency of the T allele in the DN group was significantly lower. The MMP9 -1562C allele, SBP, Cr, BUN, TG, and TC were independent risk factors for DN. All of the above suggested that the MMP9 -1562C/T polymorphism was associated with DN in Han Chinese.

The development of a quantitative and qualitative method based on UHPLC-QTOF MS/MS for evaluation paclitaxel-tetrandrine interaction and its application to a pharmacokinetic study.

Paclitaxel is a broad-spectrum anti-cancer drug by targeting microtubulin. However, multidrug resistant (MDR) makes its clinical application more difficult and results in failure of chemotherapy. Tetrandrine as a potential multidrug resistant modulator could be combined with other anti-cancer drugs. In this study, ultra-performance liquid chromatography (UHPLC) combined with quadrupole time-of-flight mass spectrometry (QTOF) was applied to simultaneously qualitative and quantitative analysis of paclitaxel for the pharmacokinetic studies while combined with tetrandrine. This method was developed based on non-target screening mode IDA (Information Dependent Acquisition). As a result, the validated range was 0.25-64ng/ml (30µl plasma) for paclitaxel. Totally 33 metabolites of paclitaxel and tetrandine were identified in vivo and in vitro. The main metabolites of PTX were dose-dependent decreased with different amounts of tetrandine co-administration no matter in vivo and in vitro, the exposure of PTX increased in pharmacokinetic study. The verified method is sensitive accurate and effective for the simultaneous determination of paclitaxel and its metabolites in blood, urine and live microsome incubation samples and it was successfully applied to evaluate the pharmacokinetics and drug-drug interaction between paclitaxel and tetrandine. Furthermore, a biosensor technology, surface plasmon resonance (SPR) analysis was applied to preliminary evaluate the competitive protein binding of multiple components. The SPR analysis indicated that the affinity between 6-hydroxy-paclitaxel and micotubulin is similar to that between paclitaxel and micotubulin, and tetrandrine also does not form a competitive combination with paclitaxel. For human, 6-hydroxy-paclitaxel is the one of main metabolites of paclitaxel, so the results suggested that tetrandine has an influence on the metabolite of paclitaxel, but tetrandine and the main metabolites of PTX probably do not affect PTX's biological targeting, the effect of its pharmacological action needs to be further studied.

Neutrophils activation can be diminished by apolipoprotein A-I.

High density lipoprotein (HDL) has anti-inflammatory function. To investigate the effects of apolipoprotein A-I (ApoA-I), the major apolipoprotein of HDL, on activated neutrophils, we stimulated neutrophils in vitro with fMLP and PMA, as a receptor-binding and a nonreceptor-binding stimuli, respectively, and incubated ApoA-I with those neutrophils. Three conditions were utilized: 1) resting neutrophils + ApoA-I (0, 2.5,5, 10 microg/mL respectively), 2) fMLP(10(-7) mol/L)-activated neutrophils + ApoA-I (0, 2.5, 5, 10 microg/mL respectively), and 3) PMA(10(-7) mol/L)-activated neutrophils + ApoA-I (0, 2.5, 5, 10 microg/mL respectively). After incubation, we measured neutrophils adhesion to fibronectin, oxidative bust (O2- and H2O2 production), degranulation (release of MPO and elastase), and L929 cell mortality which were attacked by release-out of cytokines in activated neutrophils (using MTT). Our results showed that in vitro ApoA-I inhibits fMLP- and PMA- activated neutrophil adhesion, oxidative burst, degranulation and L929 cell mortality. These inhibition effects of ApoA-I on fMLP-activated neutrophils are more powerful than that on PMA-activated neutrophils. ApoA-I has no effect on resting neutrophils. We concluded that ApoA-I could diminish the function of activated neutrophils.

High-density lipoprotein as a potential carrier for delivery of a lipophilic antitumoral drug into hepatoma cells.

AIM: To investigate the possibility of recombinant high-density lipoprotein (rHDL) being a carrier for delivering antitumoral drug to hepatoma cells. METHODS: Recombinant complex of HDL and aclacinomycin (rHDL-ACM) was prepared by cosonication of apoproteins from HDL (Apo HDL) and ACM as well as phosphatidylcholine. Characteristics of the rHDL-ACM were elucidated by electrophoretic mobility, including the size of particles, morphology and entrapment efficiency. Binding activity of rHDL-ACM to human hepatoma cells was determined by competition assay in the presence of excess native HDL. The cytotoxicity of rHDL-ACM was assessed by MTT method. RESULTS: The density range of rHDL-ACM was 1.063-1.210 g/mL, and the same as that of native HDL. The purity of all rHDL-ACM preparations was more than 92%. Encapsulated efficiencies of rHDL-ACM were more than 90%. rHDL-ACM particles were typical sphere model of lipoproteins and heterogeneous in particle size. The average diameter was 31.26+/-5.62 nm by measure of 110 rHDL-ACM particles in the range of diameter of lipoproteins. rHDL-ACM could bind on SMMC-7721 cells, and such binding could be competed against in the presence of excess native HDL. rHDL-ACM had same binding capacity as native HDL. The cellular uptake of rHDL-ACM by SMMC-7721 hepatoma cells was significantly higher than that of free ACM at the concentration range of 0.5-10 microg/mL (P<0.01). Cytotoxicity of rHDL-ACM to SMMC-7721 cells was significantly higher than that of free ACM at concentration range of less than 5 microg/mL (P<0.01) and IC50 of rHDL-ACM was lower than IC50 of free ACM (1.68 nmol/L vs 3 nmol/L). Compared to L02 hepatocytes, a normal liver cell line, the cellular uptake of rHDL-ACM by SMMC-7721 cells was significantly higher (P<0.01) and in a dose-dependent manner at the concentration range of 0.5-10 microg/mL. Cytotoxicity of the rHDL-ACM to SMMC-7721 cells was significantly higher than that to L02 cells at concentration range of 1-7.5 microg/mL (P<0.01). IC50 for SMMC-7721 cells (1.68 nmol/L) was lower than that for L02 cells (5.68 nmol/L), showing a preferential cytotoxicity of rHDL-ACM for SMMC-7721 cells. CONCLUSION: rHDL-ACM complex keeps the basic physical and biological binding properties of native HDL and shows a preferential cytotoxicity for SMMC-7721 hepatoma to normal L02 hepatocytes. HDL is a potential carrier for delivering lipophilic antitumoral drug to hepatoma cells.

Role of apolipoprotein A-I in protecting against endotoxin toxicity.

High density lipoprotein (HDL) binds lipopolysaccharide (LPS or endotoxin) and neutralizes its toxicity. We investigated the function of Apolipoprotein A-I (ApoA-I), a major apolipoprotein in HDL, in this process. Mouse macrophages were incubated with LPS, LPS+ApoA-I, LPS+ApoA-I+LFF (lipoprotein-free plasma fraction d>1.210 g/ml), LPS+HDL, LPS+HDL+LFF, respectively. MTT method was used to detect the mortality of L-929 cells which were attacked by the release-out cytokines in LPS-activated macrophages. It was found that ApoA-I significantly decreased L-929 cells mortality caused by LPS treatment (LPS vs. LPS+ApoA-I, P<0.05) and this effect became even more significant when LFF was utilized (LPS vs. LPS+ApoA-I+LFF, P<0.01; LPS vs. LPS+HDL+LFF, P<0.01). There was no significant difference between LPS+ApoA-I+LFF and LPS+HDL+LFF treatment, indicating that ApoA-I was the main factor. We also investigated in vivo effects of ApoA-I on mouse mortality rate and survival time after LPS administration. We found that the mortality in LPS+ApoA-I group (20%) and in LPS+ApoA-I+LFF group (10%) was significantly lower than that in LPS group (80%) (P<0.05, P<0.01, respectively); the survival time was (43.20 +/- 10.13) h in LPS+ApoA-I group and (46.80 +/- 3.79) h in LPS+ApoA-I+LFF group, which were significantly longer than that in LPS group (16.25 +/- 17.28) h (P<0.01). We also carried out in vitro binding study to investigate the binding capacity of ApoA-I and ApoA-I+LFF to fluorescence labeled LPS (FITC-LPS). It was shown that both ApoA-I and ApoA-I+LFF could bind with FITC-LPS, however, the binding capacity of ApoA-I+LFF to FITC-LPS (64.47 +/- 8.06) was significantly higher than that of ApoA-I alone (24.35 +/- 3.70) (P<0.01). The results suggest that: (1) ApoA-I has the ability to bind with and protect against LPS; (2) LFF enhances the effect of ApoA-I; (3) ApoA-I is the major contributor for HDL anti-endotoxin function.