Multicargo-loaded inverse opal gelatin hydrogel microparticles for promoting bacteria-infected wound healing.

Nowadays, many strategies have been developed to design biomaterials to accelerate bacteria-infected wound healing. Here, we presented a new type of multicargo-loaded inverse opal hydrogel microparticle (IOHM) for regulating oxidative stress, antibiosis, and angiogenesis of the bacteria-infected wound. The methacrylate acylated gelatin (GelMA)-based inverse opal hydrogel microparticles (IOHMs) were obtained by using the colloidal crystal microparticles as templates, and fullerol, silver nanoparticles (Ag NPs), and vascular endothelial growth factor (VEGF) were loaded in IOHMs. The developed multicargo-loaded IOHMs displayed good size distribution and biocompatibility, and when they were applied in cell culture, bacteria culture, and animal experiments, they exhibited excellent anti-oxidative stress properties, antibacterial properties, and angiogenesis. These characteristics of the developed multicargo-loaded IOHMs make them ideal for bacteria-infected wound healing.

FAR in systemic lupus erythematosus: a potential biomarker of disease activity and lupus nephritis.

The fibrinogen-to-albumin ratio (FAR), a novel inflammatory marker, has been studied in various diseases. However, the significance of FAR in systemic lupus erythematosus (SLE) has not been fully elucidated. This study was to investigate the connection between FAR and SLE. A retrospective analysis of 154 SLE patients and 77 healthy individuals was performed. The clinical and laboratory data were reviewed. Receiver operating characteristic (ROC) curves were conducted for FAR at baseline to predict disease activity and lupus nephritis (LN) in SLE patients. Pearson correlation was also applied. FAR in the SLE group was found to be significantly higher than that of the healthy control group (83.71 mg/g vs. 53.14 mg/g, P < 0.001). It was also significantly higher in patients with LN than that in patients without (107.64 mg/g vs. 67.75 mg/g, P < 0.001). The ROC curve for predicting LN showed that the area under the curve (AUC) of FAR (0.859, 95% CI 0.803-0.914) was the largest when compared to albumin (0.852, 95% CI 0.789-0.916) or fibrinogen (0.736, 95% CI 0.659-0.814) alone. In addition, FAR was a good predictor of severe disease activity in SLE (AUC = 0.721, 95% CI 0.612-0.830) and LN patients (AUC = 0.789, 95% CI 0.680-0.898). Pearson correlation analysis indicated that FAR demonstrated a strong correlation with SLE disease activity index 2000 (r = 0.4288, P < 0.001). FAR was significantly increased in SLE patients. It is a possible biomarker for disease activity and renal involvement in SLE.

LncRNA CRNDE is involved in the pathogenesis of renal fibrosis by regulating renal epithelial cell mesenchymal-epithelial transition via targeting miR-29a-3p.

Results of previous studies suggested that renal fibrosis and epithelial-mesenchymal transition (EMT) plays an important role in the process of renal fibrosis, but the underlying mechanism remains unclear. Long coding RNA (lncRNA) CRNDE has emerged as potent regulators of EMT programs, therefore, in present work, we examined the roles of LncRNA CRNDE/miR-29a-3p axis in renal fibrosis and the underlying mechanism. We found that in both renal fibrosis animal and cell models, lncRNA CRNDE was dynamically upregulated in animal models or cells by the treatment of TGF-ß. Furthermore, knockdown of CRNDE to rat significantly inhibited EMT, prevented renal fibrosis. Finally, CRNDE regulates renal fibrosis through suppression of miR-29a-3p expression. Together, our results demonstrated that CRNDE acted as a regulator of renal fibrosis via targeting miR-29a-3p. Our findings may provide a potential therapeutic target for the treatment of renal fibrosis.

Functional Characterization of Renal Tubular Epithelial Cells in Optimized Coculture Systems.

BACKGROUND: Obtaining sufficient renal tubular epithelial cells (RTCs) and maintaining the functions of RTCs are vital for developing a bioartificial renal tubule-assisted device for continuous renal replacement therapy. METHODS: We established an optimal Transwell coculture system using human primary renal proximal tubule epithelial cells (RPTECs) and bone marrow mesenchymal stem cells at different cell ratios to investigate morphological and functional changes in RTCs. Changes in cell proliferation, megalin expression, cell cycle, apoptosis, and levels of insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein-7 (BMP-7) after cell culture were investigated. RESULTS: RPTEC/BMMSC coculture at a cell ratio of 3:1 resulted in optimal morphology, function, and growth of RPTECs, in which, viability, proliferation, cytochrome P450 activity, and megalin expression in RPTECs were significantly increased compared to those in other cocultures or RPTECs alone. Additionally, IGF-1 and BMP-7 levels were significantly higher in the 3:1 RPTEC/BMMSC coculture than in the RPTECs alone. CONCLUSION: Our results demonstrate that coculture with RPTECs has great potential for use in renal replacement therapy, thereby providing fundamental information for manufacturing a bioartificial kidney.

Role of IGF-1R in epithelial-mesenchymal transdifferentiation of human peritoneal mesothelial cells.

BACKGROUND: Peritoneal fibrosis (PF) is caused by epithelial-mesenchymal transdifferentiation (EMT) in the peritoneum under high glucose (HG) conditions. The study aimed to explored the role of Insulin-like growth factor 1 receptor (IGF-1R) in the regulation of EMT in human peritoneal mesothelial cells (HPMCs). METHODS: We used HG peritoneal dialysis fluid (PDF) to induce in vivo PF in mice, and treated HPMCs with HG in vitro to stimulate EMT. RESULTS: In the mice, the higher the glucose concentration in the dialysate, the more obvious the peritoneal tissue thickening and the more that collagen was deposited. The in vitro study indicated that the expression of IGF-1R, α-SMA, vimentin was upregulated, while the expression of occludin, ZO-1, and E-cadherin was downregulated in HPMCs under HG and IGF-1R overexpression conditions. Conversely, the expression of IGF-1R, α-SMA, and vimentin was downregulated, while the expression of occludin, ZO-1, and E-cadherin was upregulated in IGF-1R-underexpressed HPMCs under HG conditions. The cell migration abilities were increased, while the cell adhesion abilities were reduced in HPMCs under HG and IGF-1R overexpression conditions. In contrast, cell migration abilities were reduced, while cell adhesion abilities were increased in IGF-1Runderexpressed HPMCs under HG conditions. CONCLUSIONS: Targeting at IGF-1R may provide novel insights into the prevention and treatment of PF.

Toll-Like Receptor 4 Is an Early and Sensitive Biomarker to Detect Acute Kidney Injury after Surgery for Type A Aortic Dissection.

Background: Acute kidney injury (AKI) is a relatively common complication after surgery for type A acute aortic dissection (ATAAD) and is associated with a poor prognosis. Preclinical models suggest that toll-like receptor 4 (TLR4) may participate in the pathogenesis of AKI. However, the correlation of serum TLR4 and post-operative AKI has not been studied in ATAAD patients. This study aimed to explore the possibility of using serum TLR4 levels to predict AKI and 30-day mortality in patients undergoing ATAAD surgery. Methods: A prospective, single-center cohort study was conducted and enrolled a total of 64 patients undergoing ATAAD surgery. The level of serum TLR4 was measured and compared before and within 24 hours after the completion of surgery. Results: Thirty-five (54.7%) patients developed AKI, including 7 (10.9%) diagnosed with severe AKI (Kidney Disease Improving Global Outcomes (KDIGO) stage 3). TLR4 levels at 0-hour,1-hour, 3-hour, and 6-hour after intensive care unit (ICU) admission were significantly different between patients with or without AKI. Further analysis showed that the difference was most significant at 0-hour after ICU admission which corresponded to an area under the curve (AUC) of 0.886 (95% confidence interval (CI), 0.800 to 0.973). For severe AKI, the AUC of TLR4 was the highest with 0.923 (0.852 to 0.995) at 1-hour after ICU admission. TLR4 levels before surgery and at 0-hour, 1-hour, as well as 3-hour after ICU admission were significantly different between survivors and non-survivors. Furthermore, we found that the serum level of TLR4 upon ICU admission could be used to predict the 30-day mortality with AUC of 0.805 (0.648 to 0.962). Conclusions: Serum TLR4 levels can be used as a biomarker to predict the occurrence of AKI and 30-day mortality in patients undergoing ATAAD surgery. Clinical Trial Registration Number: ChiCTR2200057197.

Soluble Vascular Cell Adhesion Molecule-1 Is Associated With Disease Activity in Adult-Onset Minimal Change Disease.

BACKGROUND: Cell adhesion molecules have been documented to be elevated in numerous immune inflammatory diseases. Minimal change disease (MCD) is an immune disorder. This study aimed to evaluate whether levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble E-selectin (sE-selectin) reflect disease activity in adult-onset MCD. METHODS: A sandwich enzyme-linked immunosorbent assay was used to measure the soluble adhesion molecules in 40 patients with nephrotic-range proteinuria and biopsy-proven MCD, obtained at the time of diagnosis and during remission. Thirty-five age- and sex-matched healthy volunteers served as controls. RESULTS: Patients with MCD during the active stage showed significantly higher levels of sVCAM-1 and sE-selectin when compared to controls. Moreover, sVCAM-1 had significantly positive correlations with both urine protein and serum cholesterol, and was negatively associated with serum albumin. Multiple analyses showed that serum albumin was an independent predictor of sVCAM-1. The correlations between sE-selectin and other clinical parameters were not statistically significant. At follow-up, these markers systematically decreased as the disease went into remission, but the increase in sVCAM-1 persisted even in patients obtaining complete remission for 6 months. CONCLUSIONS: Patients with active MCD had increased levels of sVCAM-1 and sE-selectin. The correlation between sVCAM-1 and proteinuria, serum albumin and cholesterol and its decline during remission indicate that sVCAM-1 is associated with disease activity.

Cross coupling of benzylammonium salts with boronic acids using a well-defined N-heterocyclic carbene-palladium(ii) precatalyst.

N-heterocyclic carbene-palladium(ii)-catalyzed cross-coupling of benzylammonium salts with arylboronic acids for the synthesis of diarylmethane derivatives via C-N bond activation has been developed. Notably, in the presence of the easily prepared and bench-stable Pd-PEPPSI precatalyst, the Csp3-N bond activation of the benzylammonium salt even proceeded smoothly in isopropanol at room temperature.

Clinical Significance of Urinary Biomarkers in Patients With Primary Focal Segmental Glomerulosclerosis.

BACKGROUND: Focal segmental glomerulosclerosis (FSGS) is often accompanied with tubulointerstitial lesion. This study aimed to assess the role of urinary biomarkers in predicting tubulointerstitial lesion and treatment response in FSGS patients. METHODS: Urinary neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), N-acetyl-ß-d-glucosaminidase (NAG) and retinol-binding protein (RBP) were measured in 32 FSGS patients and 22 patients with minimal change nephrotic syndrome. Patients with FSGS were followed up to investigate the value of these markers in predicting treatment response. RESULTS: FSGS patients had higher urinary NGAL, NAG and RBP than patients with minimal change nephrotic syndrome with comparable proteinuria. A cutoff value of 15.87ng/mL NGAL demonstrated 87.1% sensitivity and 59.1% specificity for the diagnosis of FSGS, with an area under the receiver operator characteristic curve of 0.801. In FSGS, these markers correlated significantly with the degree of acute tubulointerstitial damage but not with chronic tubulointerstitial lesion. Response to immunosuppressive therapy was significantly different in patients with KIM-1, NAG and RBP levels below and above the cutoff values. CONCLUSIONS: Urinary NGAL, KIM-1, NAG and RBP are reliable biomarkers of tubulointerstitial lesion in FSGS patients. The measurements of these markers may be useful in diagnosing FSGS, detecting acute tubulointerstitial lesion and predicting treatment response.

Renal ischemia/reperfusion-induced mitophagy protects against renal dysfunction via Drp1-dependent-pathway.

Autophagy is upregulated under stress conditions to degrade superfluous proteins and recycle damaged organelles including damaged mitochondria. However, the occurrence of mitochondrial autophagy and its contribution remain to be elucidated during renal ischemia/reperfusion injury (IRI). In this study, mitophagosomes and engulfed mitochondria were frequently observed by electron microscopy after renal IRI vs. control. Meanwhile, the increase of lipidated microtubule associated protein light chain 3 (LC3-II) and decrease of mitochondrial proteins were detected by western blot, suggesting the presence of mitophagy. Drp1 translocated to mitochondria and was phosphorylated at S616 in response to IRI. Interestingly, we found that inhibiting drp1 phosphorylation with mdivi-1 significantly suppressed IRI-induced mitophagy without affecting general autophagy. Furthermore, our results showed that downregulation of mitophagy significantly exacerbated cell apoptosis and markedly aggravated kidney dysfunction induced by IRI. Taken together, these data indicate that mitophagy was activated via Drp1-dependent pathway and such mitophagic clearance of damaged mitochondria protects cells from IRI-induced apoptosis.

Vitamin D supplementation improves endothelial dysfunction in patients with non-dialysis chronic kidney disease.

PURPOSE: Hypovitaminosis D is common in chronic kidney disease (CKD) and is associated with endothelial dysfunction and cardiovascular events. This study aimed to investigate the effects of vitamin D supplementation on endothelial dysfunction in non-dialysis CKD patients. MATERIALS AND METHODS: Seventy-one non-dialysis CKD patients with low vitamin D (serum 25(OH)D < 30 ng/mL) were recruited. Patients received oral cholecalciferol 50,000 units once a week for 12 weeks. Changes in endothelial function by brachial artery flow-mediated dilation (FMD), soluble vascular cell adhesion molecule-1 (sVCAM-1), and sE-selectin were studied. RESULTS: There was a significant increase in serum levels of 25(OH)D after cholecalciferol supplementation (33.7 ± 12.1 vs. 13.2 ± 5.4 ng/mL, P < 0.001). Multivariable regression analysis showed that higher proteinuria (ß = - 0.548, P < 0.001) and lower levels of 25(OH)D (ß = 0.360, P < 0.001) at baseline were related to lower 25(OH)D level after supplementation. FMD increased significantly from 4.4 ± 1.3 to 5.1 ± 1.5% (P < 0.001), and soluble endothelial biomarkers decreased: sVCAM-1 from 926.9 ± 158.0 to 867.0 ± 129.0 ng/mL (P < 0.001), and sE-selectin 69.7 ± 15.8 to 63.3 ± 14.7 ng/mL (P < 0.001). CONCLUSIONS: Vitamin D supplementation can improve endothelial dysfunction in pre-dialysis CKD patients.

Effects of continuous renal replacement therapy on serum cytokines, neutrophil gelatinase-associated lipocalin, and prognosis in patients with severe acute kidney injury after cardiac surgery.

The aim of our study was to evaluate the effect of continuous renal replacement therapy (CRRT) on serum cytokines, neutrophil gelatinase-associated lipocalin (NGAL), and prognosis in patients with severe acute kidney injury (AKI) following cardiac surgery. A total number of 153 patients with severe AKI following cardiac surgery were treated with CRRT. They were divided into the survival and non-survival groups. Clinical data from these two groups before and after CRRT were recorded and analyzed. It was found that the number of impaired organs, MODS and APACHE II scores were significantly higher in the non-survival group than those in the survival group before CRRT. After CRRT, MODS and APACHE II scores decreased significantly. The post-CRRT levels of serum TNF-α and IL-6 were significantly decreased. After CRRT, serum NGAL decreased in the two groups, but the levels were higher in the non-survival group than those in the survival group. MODS and APACHE II scores could be used to evaluate the severity of AKI in patients after cardiac surgery. CRRT is an effective treatment for these patients and high levels of TNF-α, IL-6, and NGAL are associated with a poor prognosis in these patients.

Serum neutrophil gelatinase-associated lipocalin and proinflammatory cytokines in pigs with septic versus non-septic acute kidney injury.

BACKGROUND: The aim of this study was to establish a composite pig model with sepsis and ischemic acute kidney injury (AKI) and to investigate the differences in serum neutrophil gelatinase-associated lipocalin (NGAL) and proinflammatory cytokines in septic and non-septic AKI. METHODS: Seventeen healthy hybridized pigs (weighed 26.97 ± 2.26 kg) were randomly divided into two groups. Group A (n = 12) served as the septic AKI model which received cecal ligation and puncture, resulting in abdominal infection plus clamping of renal artery (CRA). Group B (n = 5) received CRA only. Vital signs and the functions of the main organs were observed. Serum NGAL, TNF-α, and IL-6 were measured at 0, 4, 8, 24, and 48 h after surgical admissions. RESULTS: Septic AKI model was successfully induced, which manifested as multiple organ dysfunction syndrome, including AKI, liver dysfunction, progressive decline of cardiac function and abnormal pulmonary function. Apparent pathological changes were found in kidney, liver, lung and small intestine of group A. The proinflammatory cytokines in Group A were significantly higher than those in Group B at different time points (P < 0.05). In Group A, serum concentrations of TNF-α reached the peak at 8 h, while IL-6 levels dramatically increased at 24 h. There was a significant difference in serum NGAL between Group A and B at 8 h (P < 0.05). CONCLUSIONS: Septic AKI animals have higher serum NGAL compared with non-septic AKI animals. Monitoring the activities of TNF-α, NGAL and IL-6 would make great contributions in discovering sepsis and evaluating the severity of sepsis.

Hypoxia inducible factor-1 alpha promotes mesangial cell proliferation in lupus nephritis.

BACKGROUND: Evidence has accumulated that hypoxia plays a significant role in the pathogenesis and progression of both acute renal injury and chronic renal disease. However, little was known about the effects of hypoxia on lupus nephritis (LN). In the current study, we investigated the expression of hypoxia inducible factor-1 alpha (HIF-1α) in LN. METHODS: Renal biopsies from 22 LN patients and 20 patients with renal carcinoma were obtained. In situ HIF-1α expression was examined by immunohistochemical staining, and the relationship between HIF-1α and clinical/pathological features was analyzed. HIF-1α expression in kidney from both MRL/lpr and C57BL/6 mice was detected by immunohistochemical technology. Dimethyloxaloylglycine (DMOG), an inhibitor of HIF-degrading prolylhydroxylases, was utilized to prevent HIF-1α degradation in mouse mesangial cells (MCs). After DMOG treatment, the proliferation and apoptosis rates of mouse MCs were determined. RESULTS: LN patients showed larger amounts of HIF-1α in both glomerular and tubulointerstitial areas. The levels of intraglomerular HIF-1α were closely associated with renal pathology activity index and clinical manifestations in LN patients. In MRL/lpr mice, intraglomerular HIF-1α-positive cells were also significantly increased. Interestingly, the levels of HIF-1α positively correlated with cell density in glomerulus in both LN patients and MRL/lpr mice. Upon treatment with DMOG, the proliferation of MCs was upregulated, and apoptosis was downregulated. CONCLUSION: HIF-1α is highly expressed in both glomerular and tubulointerstitial tissues in LN, especially in proliferative LN. HIF-1α may promote MCs growth through the induction of proliferation and inhibition of apoptosis, and hence plays an important role in the pathogenesis of LN.

Improvement of cytokine response and survival time by bioartificial kidney therapy in acute uremic pigs with multi-organ dysfunction.

OBJECTIVE: To explore whether bioartificial kidney (BAK) ameliorates cytokine response and biochemical indices, and prolongs the survival time in acute uremic pigs with multiple organ dysfunction syndrome (MODS). METHODS: Hybridized pigs suffering from MODS and acute renal failure (ARF) were treated with BAK (Group A, n=6) or sham-BAK containing no cells (Group B, n=6), or received no treatment (Group C, n=5). Data on blood pressure, hepatic and renal function, IL-10, TNF-α, arterial blood gas, and survival time of all the pigs was recorded. RESULTS: Mean arterial pressure (MAP, mmHg) responded more rapidly and reached higher values in Group A (91.82 ± 5.73) compared with Groups B and C at 24 hours (p<0.01). The peak level of serum IL-10 (pg/mL) in Group A (249.57 ± 43.51) was significantly higher than in Groups B and C (132.06 ± 17.53, 104.25 ± 13.42, p<0.01). Serum TNF-α level (pg/mL) in Group A dropped gradually to 402.91 ± 32.47 at 24 hours, and showed a significant discrepancy compared with those before treatment (537.16 ± 38.45) and Group B (512.94 ± 19.5, p<0.05). There was no difference in plasma endotoxin and serum IL-6 between pre-treatment and post-treatment in Groups A and B. BAK treatment, however, resulted in a significant decline in IL-6/IL-10 ratios. The average survival time (hours) in Group A (113.01 ± 14.32) was significantly longer, prolonged by 35.93% and 63.90% compared to Groups B and C (p<0.01), respectively. CONCLUSIONS: The addition of renal tubule cell therapy to hemofiltration in an acutely uremic animal model with MODS altered systemic cytokine balance, ameliorated MAP, and prolonged survival time.