Downregulation of miR-574-5p inhibits HK-2 cell viability and predicts the onset of acute kidney injury in sepsis patients

BackgroundIncreased levels of microRNA-574-5p (miR-574-5p) have been found to be associated with increased survival of septic patients, indicating the potential role of miR-574-5p in protecting against septic progression and complications. Acute kidney injury (AKI) is one of the most common and serious complications of sepsis. Therefore, the aim of this study was to test these hypotheses: (1) in a renal cell culture line (HK-2), upregulated expression of miR-574-5p increases, and downregulated expression of miR-574-5p decreases cell viability, and (2) serum levels of miR-574-5p from patients with sepsis and AKI are lower than those of patients with sepsis but no AKI.MethodsThe expression of miR-574-5p was regulated by cell transfection in HK-2 cells, and HK-2 cell viability was measured using the Cell Counting Kit-8. Serum miR-574-5p expression was analyzed using qRT-PCR. The predictive value of miR-574-5p for AKI onset was evaluated using the receiver operating characteristic curve and logistic regression analysis.ResultsThe overexpression of miR-574-5p promoted HK-2 cell viability. Fifty-eight sepsis patients developed AKI, who had significantly lower miR-574-5p expression. miR-574-5p expression was decreased with AKI stage increase and correlated with kidney injury biomarker and had relatively high accuracy to predict AKI occurrence from sepsis patients.ConclusionOverexpression of miR-574-5p in cultured HK-2 cells increases cell viability and knocked-down expression of miR-574-5p decreases cell viability. Consistently, septic patients with AKI were found to have less upregulation of miR-574-5p expression compared to septic patients without AKI. Thus, serum miR-574-5p may provide a novel biomarker for septic AKI.     

miR-34b-5p promotes renal cell inflammation and apoptosis by inhibiting aquaporin-2 in sepsis-induced acute kidney injury

ObjectiveThis study was designed to uncover the mechanism of miR-34b-5p-mediated aquaporin-2 (AQP2) in sepsis-induced injury using human renal tubular epithelial cells (HK-2).MethodsSerum levels of miR-34b-5p, TNF-α, IL-1β, IL-6, serum creatinine (SCr), and blood urea nitrogen (BUN) in septic patients with acute kidney injury (AKI) and healthy controls were detected. Lipopolysaccharide (LPS) was used to induce sepsis in HK-2 cells. LPS-induced HK-2 cells were transfected with miR-34b-5p inhibitor, miR-34b-5p mimic, pcDNA3.1-AQP2, si-AQP2, miR-34b-5p inhibitor + si-NC, or miR-34b-5p inhibitor + si-AQP2. The expressions of miR-34b-5p, AQP2, Bax, Bcl-2, cleaved caspase-3, TNF-α, IL-1β, and IL-6 in HK-2 cells were detected. TUNEL staining revealed the apoptosis of HK-2 cells. Dual-luciferase reporter assay verified the binding between miR-34b-5p and AQP2.ResultsThe expression of miR-34b-5p and the inflammatory responses were augmented in septic AKI patients. miR-34b-5p was up-regulated and AQP2 was down-regulated in LPS-induced HK-2 cells. miR-34b-5p inhibition or AQP2 overexpression ameliorated apoptosis and inflammation in LPS-induced HK-2 cells. In contrast, overexpressing miR-34b-5p deteriorated LPS-induced injury in HK-2 cells. AQP2 was a downstream target of miR-34b-5p. AQP2 silencing abolished the suppressive effects of miR-34b-5p inhibition on LPS-induced apoptosis and inflammatory response in HK-2 cells.ConclusionmiR-34b-5p inhibits AQP2 to promote LPS-induced injury in HK-2 cells.     

The miR-26a-5p/IL-6 axis alleviates sepsis-induced acute kidney injury by inhibiting renal inflammation

Sepsis-induced acute kidney injury (AKI) is a common and life-threatening complication in hospitalized and critically ill patients and has unacceptable morbidity and mortality rates. However, effective approaches for the diagnosis and treatment of septic AKI are still lacking. Here, we demonstrated significant increases in the miR-26a-5p levels in renal tubular cells of LPS-induced septic AKI models both in vivo and in vitro. Mechanistically, we provided evidence of the involvement of NF-κB in miR-26a-5p induction. LPS treatment of renal tubular cells led to the activation of NF-κB, and inhibition of NF-κB by TPCA-1 prevented the induction of miR-26a-5p. These results indicated that NF-κB was a key upstream factor for the induction of miR-26a-5p in septic AKI. Anti-miR-26a-5p enhanced the expression of IL-6 at both the protein and mRNA levels following LPS treatment. Furthermore, our luciferase microRNA target reporter assay verified that IL-6 is a direct target of miR-26a-5p. Blocking miR-26a-5p promoted renal inflammation and worsened kidney injury. Thus, our study indicated that the miR-26a-5p/IL-6 axis can alleviate sepsis-induced acute kidney injury by inhibiting renal inflammation. This mechanism may represent a therapeutic target for septic AKI.     

MiR-702-3p inhibits the inflammatory injury in septic H9c2 cells by regulating NOD1

BackgroundCardiac insufficiency is a common complication of sepsis and septic shock and is the most common cause of death in critically ill patients. Recent studies have found that microRNAs (miRNAs) play a potential role in sepsis as markers, but little is known about their functional effects on sepsis-induced cardiomyopathy (SIC).ObjectiveThis study is designed to explore the possible role and underlying mechanisms of miR-702-3p in septic cardiomyopathy.MethodsAs expected, H9c2 cells were induced with lipopolysaccharide (LPS) to construct the model of septic cardiomyopathy. The expression of miR-702-3p was detected by qRT-PCR assay and those of IL-1β, IL-6 and TNF-α by ELISA assay. The viability, proliferation and apoptosis of LPS-treated H9c2 cells were determined by CCK-8, EdU, flow cytometry and western blot assays. Moreover, Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) was predicted and confirmed as a direct target of miR-702-3p by TargetScan, miRwalk and miRDB prediction and dual-luciferase reporter gene assays.ResultsWhile LPS can weaken the viability of H9c2 cells, miR-702-3p enhances that of LPS-treated H9c2 cells by inhibit the expressions of TNF-α, IL-6, IL-1β. We found NOD1 is a target gene of miR-702-3p, and over-expression of NOD1 restores the inhibitory effects of miR-702-3p on the LPS-treated H9c2 cells.ConclusionMiR-702-3p played an important role in the pathogenesis of sepsis cardiomyopathy via targeting NOD1, suggesting that miR-702-3p may be a potential new target for the treatment of SIC.     

Urinary micro-RNA expressions and protein concentrations may differentiate bladder cancer patients from healthy controls

Purpose

To determine expression differences of urine exosomal miR-19b1-5p, 21-5p, 136-3p, 139-5p, 210-3p and concentration differences of urinary BLCA-4, NMP22, APE1/Ref1, CRK, VIM between bladder cancer, follow-up patients, and control samples, to evaluate diagnostic importance of these differences and establish a diagnostic panel for bladder cancer.

Methods

Urine samples of 59 bladder cancer patients, 34 healthy controls, and 12 follow-up patients without recurrence were enrolled to this study. Real-time PCR and ELISA were performed to determine urine exosomal miR-19b1-5p, 21-5p, 136-3p, 139-5p, 210-3p expressions and urinary BLCA-4, NMP22, APE1/Ref1, CRK, VIM, creatinine concentrations. Logistic regression analyses were performed to determine the diagnostic panel, the sensitivity, and specificity of the panel assessed by the ROC curve analysis. p values?<?0.05 were considered statistically significant.

Results

In bladder cancer risk groups, mir-139, -136, -19 and 210 expressions or positivity were found to be different and concentrations of urinary Ape1/Ref1, BLCA4, CRK, and VIM increased by twofold on average compared to healthy controls. Logistic regression and ROC analyses revealed that panel could differentiate bladder cancer patients from healthy controls with 80% sensitivity and 88% specificity (AUC?=?0.899), low-risk patients from controls with 93% sensitivity and 95.5% specificity (AUC?=?0.976). Despite the low number of samples, our findings suggest that urine exosomal miR-19b1-5p, 136-3p, 139-5p expression, and urinary APE1/Ref1, BLCA-4, CRK concentrations are promising candidates in terms of bladder cancer diagnosis.

Conclusions

Although our panel has great sensitivity for early detection of BC, it needs to be validated in larger populations.

     

Septic acute kidney injury in critically ill patients – a single-center study on its incidence,clinical characteristics,and outcome predictors

Purpose The objective of this study is to examine the incidence, clinical characteristics, and outcome (90-day mortality) of critically ill Chinese patients with septic AKI. Methods Patients admitted to the ICU of a regional hospital from 1 January 2011 to 31 December 2013 were included, excluding those on chronic renal replacement therapy. AKI was defined using KDIGO criteria. Patients were followed till 90 days from ICU admission or death, whichever occurred earlier. Demographics, diagnosis, clinical characteristics, and outcome were analyzed. Results In total, 3687 patients were included and 54.7% patients developed AKI. Sepsis was the most common cause of AKI (49.2%). Compared to those without AKI, AKI patients had higher disease severity, more physiological and biochemical disturbance, and carried significant co-morbidities. Ninety-day mortality increased with severity of AKI (16.7, 27.5, and 48.3% for KDIGO stage 1, 2, and 3 AKI, p?<?0.001). Full renal recovery was achieved in 71.6% of AKI patients. Compared with non-septic AKI, septic AKI was associated with higher disease severity and required more aggressive support. Non-recovery of renal function occurred in 2.5% of patients with septic AKI, compared with 6.4% in non-septic AKI (p?<?0.001). Cox regression analysis showed that age, emergency ICU admission, post-operative cases, admission diagnosis, etiology of AKI, disease severity score, mechanical ventilation, vasopressor support, and blood parameters (like albumin, potassium and pH) independently predicted 90-day mortality. Conclusions AKI, especially septic AKI is common in critically ill Chinese patients and is associated with poor patient outcome. Etiology of AKI has a significant impact on 90-day mortality and may affect renal outcome.     

Assessing the influence of acute kidney injury on the mortality in patients with acute myocardial infarction: a clinical trail

Objectives: Acute kidney injury (AKI) increases the risk of death following acute myocardial infarction (AMI). In this current study, we tried to understand the role of newly KDIGO defined AKI in AMI-induced early and late mortality.

Methods: We retrospectively analyzed the clinical data of AMI patients (totaling 1371 cases) from the hospital’s computer database. And AKI was defined based on the KDIGO criteria but GFR or urinary output assessment was not used. Subsequently, we compared the association of AKI with 30-day and 30-day to 5-year all-cause mortality, using multivariate COX regression analysis with two models.

Results: We observed the development of AKI in 410 (29.9%) patients during the hospital stay. The 30-day and 30-day to 5-year mortality rates were 5.6% and 11.3%, respectively, in 1371 AMI patients. Further, adjusted Cox regression analysis based on model 1 revealed that AKI severity was an independent risk factor of 30-day mortality, while AKI Stage 3 was an independent predictor of 30-day to 5-year mortality. Adjusted Cox regression analysis based on model 2 revealed that normal baseline renal function with AKI and impaired renal function with AKI were independent risk factors of 30-day mortality, while normal baseline renal function with AKI and impaired renal function with AKI were identified to be independent predictors of 30-day to 5-year mortality.

Conclusions: Whether the baseline renal function decreased or not, AKI strongly correlated with short- and long-term all-cause mortality in patients with AMI. Specifically, the short-term mortality of AMI patients increased with more severe AKI.     

Implications of dynamic changes in miR-192 expression in ischemic acute kidney injury

Purpose

Ischemia–reperfusion injury (IRI) is a major cause of acute kidney injury (AKI) with poor outcomes. While many important functions of microRNAs (miRNAs) have been identified in various diseases, few studies reported miRNAs in acute kidney IRI, especially the dynamic changes in their expression and their implications during disease progression.

Methods

The expression of miR-192, a specific kidney-enriched miRNA, was assessed in both the plasma and kidney of IRI rats at different time points after kidney injury and compared to renal function and kidney histological changes. The results were validated in the plasma of the selected patients with AKI after cardiac surgery compared with those matched patients without AKI. The performance characteristics of miR-192 were summarized using area under the receiver operator characteristic (ROC) curves (AUC-ROC).

Results

MiRNA profiling in plasma led to the identification of 42 differentially expressed miRNAs in the IRI group compared to the sham group. MiR-192 was kidney-enriched and chosen for further validation. Real-time PCR showed that miR-192 levels increased by fourfold in the plasma and decreased by about 40% in the kidney of IRI rats. Plasma miR-192 expression started increasing at 3 h and peaked at 12 h, while kidney miR-192 expression started decreasing at 6 h and remained at a low level for 7 days after reperfusion. Plasma miR-192 level in patients with AKI increased at the time of ICU admission, was stable for 2 h and decreased after 24 h. AUC-ROC was 0.673 (95% CI: 0.540–0.806, p = 0.014).

Conclusions

Plasma miR-192 expression was induced in a time-dependent manner after IRI in rats and patients with AKI after cardiac surgery, comparably to the kidney injury development and recovery process, and may be useful for the detection of AKI.

     

The optimal timing of continuous renal replacement therapy for patients with sepsis-induced acute kidney injury

Purpose

High mortality in the intensive care unit (ICU) is probably associated with sepsis-induced acute kidney injury (AKI). The aim of this study is to explore which stage of AKI may be the optimal timing for continuous renal replacement therapy (CRRT).

Methods

A retrospective analysis of 160 critically ill patients with septic AKI, treated with or without CRRT was performed in Binzhou medical college affiliated hospital ICU. The parameters including 28-days mortality rate, renal recovery, ventilation time and ICU stay between CRRT group and control group were assessed.

Results

Renal recovery, defined as independence from dialysis at discharge, was documented for 64/76 (84.2 %) of the surviving patients (48.1 % of total subjects included in the study). The mortality rate increased proportionally with acute kidney injury Network stages in CRRT subgroups (P = 0.001) and control groups (P = 0.029). CRRT initiation at stage 2 of AKI significantly reduced the 28-day mortality (P = 0.048) and increased the 28-day survival rate (P = 0.036) compared with those in control group. In addition, the ICU stay and ventilation time were shorter in CRRT group than that of control group in stage 2 of AKI.

Conclusion

The stage 2 AKI might be the optimal timing for performing CRRT.     

Clinical characteristics of sepsis-induced acute kidney injury in patients undergoing continuous renal replacement therapy

Objective: The aim of this study was to investigate the clinical characteristics of sepsis-induced acute kidney injury (AKI) in patients undergoing continuous renal replacement therapy (CRRT).

Methods: From 2011 to 2015, we enrolled 340 patients who were treated with CRRT for sepsis at the Presbyterian Medical Center. In all patients, CRRT was performed using the PRISMA platform. We divided these patients into two groups (survivors and non-survivors) according to the 28-day all-cause mortality. We compared clinical characteristics and analyzed the predictors of mortality.

Results: The 28-day all-cause mortality was 62%. Survivors were younger than non-survivors and had higher platelet counts (178?±?101?×?10³/mL vs. 134?±?84?×?10³/mL, p?p?p?p?0.05?mL/kg/h (66% vs. 86%, p?=?.001) in the first day. In a multivariate logistic regression analysis, age, platelet count, RDW score, APACHE II score, serum creatinine level, and a urine output of <0.05?mL/kg/h the first day were prognostic factors for the 28-day all-cause mortality.

Conclusion: Age, platelet count, APACHE II score, RDW score, serum creatinine level, and urine output the first day are useful predictors for the 28-day all-cause mortality in sepsis patients requiring CRRT.     

Simvastatin improves sepsis-induced mortality and acute kidney injury via renal vascular effects

Acute kidney injury (AKI) occurs in about half of patients in septic shock and the mortality of AKI with sepsis is extremely high. An effective therapeutic intervention is urgently required. Statins are 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors that also have pleiotropic actions. They have been reported to increase the survival of septic or infectious patients. But the effect of simvastatin, a widely used statin, on sepsis-induced AKI is unknown. The effects of simvastatin and tumor necrosis factor (TNF)-alpha neutralizing antibody were studied in a clinically relevant model of sepsis-induced AKI using cecal ligation and puncture (CLP) in elderly mice. Simvastatin significantly improved CLP-induced mortality and AKI. Simvastatin attenuated CLP-induced tubular damage and reversed CLP-induced reduction of intrarenal microvascular perfusion and renal tubular hypoxia at 24 h. Simvastatin also restored towards normal CLP-induced renal vascular protein leak and serum TNF-alpha. Neither delayed simvastatin therapy nor TNF-alpha neutralizing antibody improved CLP-induced AKI. Simvastatin improved sepsis-induced AKI by direct effects on the renal vasculature, reversal of tubular hypoxia, and had a systemic anti-inflammatory effect.     

Circulating miR-200c Levels Significantly Predict Response to Chemotherapy and Prognosis of Patients Undergoing Neoadjuvant Chemotherapy for Esophageal Cancer

Background

There is increasing evidence that microRNA expression in cancer tissue is useful for predicting the prognosis of patients with cancer. However, the relationship between the levels of circulating microRNAs and response to chemotherapy and prognosis remains unclear in esophageal cancer.

Methods

We measured the serum levels of miR-21, miR-145, miR-200c, and let-7c by quantitative RT-PCR in 64 patients with esophageal cancer who underwent neoadjuvant chemotherapy.

Results

The serum levels of miR-21, miR-145, miR-200c, and let-7c in esophageal cancer patients were significantly higher than those in healthy volunteers. High expression of miR-200c correlated significantly with poor response to chemotherapy (p = 0.0211). There was no significant relationship between chemosensitivity and the levels of miR-21, miR-145, and let-7c. High expression of miR-200c was associated with shortened progression-free survival (p = 0.0076), but there was no significant relationship between prognosis and the expression of miR-21, miR-145, and let-7c. Multivariate analysis identified miR-200c expression as the most valuable prognostic factor for patients with esophageal cancer who receive neoadjuvant chemotherapy.

Conclusions

The serum level of miR-200c can be useful for predicting the response to chemotherapy and the prognosis of patients with esophageal cancer who receive neoadjuvant chemotherapy.

     

瞬时受体电位类香草醛5和骨桥蛋白在泌尿系统结石患者中的表达及其与微小RNA-22-3p的关系

目的检测瞬时受体电位类香草醛5(TRPV5)和骨桥蛋白(OPN)在泌尿系统结石患者中的表达及其与微小核糖核酸-22-3p(miR-22-3p)的关系。方法荧光定量反转录-聚合酶链式反应(RT-qPCR)检测健康体检者、非泌尿系统结石患者和泌尿系统结石血清TRPV5、OPN和miR-22-3p表达并分析其相关性。将模型+miR-con组(转染miR-con)、模型+miR-22-3p组(转染miR-22-3p)转染至HK-2细胞并草酸钙结石晶体溶液(100 mg/L)处理。流式细胞仪检测细胞凋亡;蛋白质印迹法(Western blot)检测抗裂解的半胱氨酸天冬氨酸蛋白酶-3(cleaved Caspase-3)、B淋巴细胞瘤-2(bcl-2)和bcl-2相关X蛋白(bax)表达。两组间比较采用t检验,多组间比较采用单因素方差分析联合SNK-q检验。采用Pearson相关性分析法分析相关性。结果与健康体检者或非泌尿系统结石患者比较,泌尿系统结石患者血清TRPV5和miR-22-3p表达显著降低(TRPV5分别为0.66±0.15、1.06±0.26、0.77±0.21,miR-22-3p分别为0.62±0.18、1.00±0.23、0.83±0.19),OPN表达显著升高(2.78±0.49、1.07±0.21、1.60±0.20),差异有统计学意义(F=43.908、300.788、41.410,P<0.05)。泌尿系统结石患者血清中miR-22-3p的表达量与TRPV5的表达呈正相关(r=0.52,P<0.05),与OPN的表达呈负相关(r=-0.69,P<0.05)。与对照组比较,模型组HK-2细胞中TRPV5和miR-22-3p表达显著降低(TRPV5为0.43±0.07比0.98±0.10,miR-22-3p为0.72±0.16比1.01±0.13),OPN表达显著升高(3.24±0.36比1.02±0.12),差异有统计学意义(t=13.517、75.794和17.551,P<0.05),细胞凋亡率[(17.37±2.09)%比(3.78±0.49)%]、cleaved Caspase-3(0.42±0.06比0.19±0.05)和bax(0.68±0.07比0.28±0.06)蛋白质表达均显著升高,bcl-2蛋白表达显著降低(0.14±0.03比0.31±0.05),差异有统计学意义(t=21.431、17.233、13.655、31.782,P<0.05)。与模型+miR-con组比较,模型+miR-22-3p组细胞上述检测指标均发生显著的负向调控。结论TRPV5和OPN在泌尿系统结石患者中的表达与miR-22-3p存在明显的相关性。     

Effects of the diagnostic window and duration of acute kidney injury on 1-year mortality in elderly patients: a single-center retrospective study

Background

We evaluated the prognostic impact of AKI duration on the 1-year mortality rate in elderly patients diagnosed based on the 48-hour and 7-day changes in serum creatinine (Scr) levels recommended by the Kidney Disease: Improving Global Outcomes (KDIGO) guidelines.

Methods

This retrospective study was conducted from 2007 to 2018 on elderly patients in the Geriatric Department of the Chinese PLA General Hospital. Based on the two diagnostic criteria in the KDIGO guidelines, the patients were divided into a 48-hour diagnostic window and a 7-day diagnostic window group, and into transient AKI (lasting 1–2 days) and persistent AKI (lasting 3–6 days, and?≥?7 days) based on the time at which the Scr level returned to the baseline value. The primary outcome was the 1-year mortality rate after AKI.

Results

In total, 688 patients were enrolled, including 367 (53.3%) with a 48-hour and 321 (46.7%) with a 7-day diagnostic window. Of the 688 patients, in the 48-hour window group, 12.0% had transient AKI, 31.1% had lasting 3–6 days, and 56.9% had lasting?≥?7 days; in the 7-day window group, 5.3% had transient AKI, 24.0% had lasting 3–6 days, and 70.7% had lasting?≥?7 days. Overall, 332 patients (33.6%) died within 1 year, including 189 (51.5%) in the 48-hour and 143 (44.5%) in the 7-day diagnostic window group. After adjusting for multiple covariates, AKI duration was associated with a significantly higher 1-year mortality rate (3–6 days: HR?=?3.535; 95% CI?=?1.685–7.417, P?=?0.001;?≥?7 days: HR?=?2.400; 95% CI?=?1.152–5.001, P?=?0.019) in the 48-hour diagnostic window group, but it did not differ in the 7-day diagnostic window group (P?=?0.452).

Conclusions

Persistent AKI was common in elderly hospitalized patients, accounting for 88% and 95% of patients with 48-hour and 7-day diagnostic windows, respectively. Moreover, AKI duration was associated with different clinical outcomes depending on the diagnostic window. Further studies should focus on the mechanism underlying the relationship of AKI outcomes with diagnostic criteria.

     

MiR-30b-3p and miR-126-3p of urinary extracellular vesicles could be new biomarkers for prostate cancer

BackgroundExtracellular vesicles (EVs) including exosomes are present in blood, urine, and saliva and contain proteins, microRNAs, and messenger RNAs. We investigated microRNAs in urinary EVs to discover new biomarkers of prostate cancer (PCa).MethodsWe isolated EVs from urine obtained following digital rectal examination (DRE) of 14 men with elevated levels of serum prostate-specific antigen (PSA) [negative biopsy (n=4) and PCa with Gleason scores of 6 (n=3), 7 (n=3), and 8–9 (n=4)]. MicroRNAs extracted from EVs were analyzed by microRNA microarray.ResultsMicroRNAs miR-30b-3p and miR-126-3p were identified as being overexpressed in urinary EVs of the PCa patients versus the biopsy-negative men, but no microRNAs were associated with the Gleason score. In the independent cohort as well, these two microRNAs were overexpressed in urinary EVs from the PCa patients versus the negative-biopsy men. Logistic regression analysis adjusted by age and PSA showed that these two microRNAs were significantly associated with the prediction of PCa in biopsy specimens. Sensitivity and specificity of miR-30b-3p and miR-126-3p for the prediction of PCa were 46.4% and 88.0% and 60.7% and 80.0%, respectively, which were better than those of serum PSA (53.5% and 64.0%, respectively).ConclusionsMiR-30b-3p and miR-126-3p in urinary EVs could be potential biomarkers of PCa.     

PenKid measurement at admission is associated with outcome in severely ill burn patients

BackgroundProenkephalin A 119–159 (penKid) has been proposed as a sensitive biomarker of renal function. This study evaluated the association of concentrations of plasma penKid with death and risk of acute kidney injury (AKI) in severely ill burn patients.MethodsA prospective observational study in two centers with severely ill adult burn patients was conducted. The inclusion criteria were total body surface area (TBSA) burns >15%, with burn injury occurring <72 h before intensive care unit (ICU) admission and plasma sample taken at admission. The primary endpoint was 90-day mortality. The secondary endpoints were AKI and a combined endpoint of 90-day mortality and/or AKI. Mortality was also evaluated in the sub-group of patients with sub-clinical AKI, defined as a patient without AKI but with elevated penKid.ResultsA total of 113 consecutive patients were enrolled. The median age was 48 years (Interquartile range [IQR] 33–64), the median burn TBSA was 35% (IQR 25–53), and 90-day mortality was 31.9%. Thirty-one percent of the patients had AKI, and 41.6% of patients had the combined endpoint. There was a stepwise decrease in survival from patients without AKI, sub-AKI, and with AKI (survival rate 90.0% [95% CI 82.7–97.9], 66.7% [95% CI 48.1–92.4], and 31.4% [95% CI 19.3–51.3], respectively, p < 0.001). Plasma penKid concentration was significantly higher in non-survivors compared to survivors (86.9 pmol/L [IQR 53.3–166.1] versus 52.9 pmol/L [IQR 37.1–70.7]; p = 0.0001) and in patients with AKI compared to patients without AKI (86.4 pmol/L [IQR 56.5–153.4] versus 52.5 pmol/L [IQR 35.5–71.2]; p < 0.001). Penkid provided added value on top of serum creatinine (Screat) and Sepsis Related Organ Failure Assessment (SOFA) scores to predict 90-day mortality (combined c-index of 0.738 versus 0.707; p = 0.024 and 0.787 versus 0.752; p < 0.001).ConclusionsPlasma penKid concentration at admission was associated with an increased risk of death in burn patients. PenKid has additional prognostic value on top of Screat and SOFA to predict 90-day mortality.     

Clinical significance of serum miR-129-5p in patients with diabetes mellitus presenting macrovascular complications

BACKGROUNDDiabetic macrovascular complications (DMCs) are the most common complications encountered during the course of diabetes mellitus (DM) with extremely high mortality rates. Therefore, there is an urgent need to identify specific and sensitive biomarkers for the early diagnosis of DMCs.AIMTo investigate the expression and significance of serum miR-129-5p in patients with DM and macrovascular complications.METHODSSerum samples were collected from 36 healthy controls, 58 patients with DM presenting no macrovascular complications, and 62 patients with DMCs. The expression of miR-129-5p was detected using quantitative real-time polymerase chain reaction. Pearson’s correlation assay was performed to analyze the correlation between serum miR-129-5p levels and clinical indicators. Receiver operator characteristic (ROC) analysis was conducted to analyze the diagnostic value of serum miR-129-5p in patients with DM or DMCs.RESULTSThere was a 4.378-fold and 7.369-fold increase in serum miR-129-5p expression in the DM (5.346 ± 0.405) and DMCs (8.998 ± 0.631) groups, respectively (P < 0.001), compared with the control group (1.221±0.090). In addition, the expression of serum miR-129-5p in patients with DMCs was higher than that in patients with DM, revealing a 1.683-fold increase (P < 0.001). Additionally, serum miR-129-5p expression significantly correlated with smoking history, disease duration, and glycated hemoglobin (HbA1c) in patients with DMCs (P < 0.001). The area under the ROC curve (AUC) of miR-129-5p as a serum marker was 0.964 (95% confidence interval [CI]: 0.930-0.997, P < 0.001) in distinguishing between patients with DM and healthy controls, whereas the AUC of miR-129-5p as a serum marker was 0.979 (95%CI: 0.959-0.999, P < 0.001) in distinguishing between patients with DMCs and healthy controls. CONCLUSIONElevated serum miR-129-5p expression levels correlate with the development of DMCs and can be utilized as a novel early diagnostic biomarker for DM combined with macrovascular complications.     

miR-18a-3p在肝癌中的表达及其调控ADCY1表达对肝癌细胞侵袭及增殖的影响

背景与目的 研究表明多种microRNA（miRNA）可能在肝癌的发生发展中发挥重要作用,其作用机制仍值得进一步研究和探讨。因此,本研究从已报道的肝癌差异表达miRNA中进一步筛选关键miRNA,并验证和探讨其作用机制。方法 从已发表的研究中筛选出肝癌组织及肝癌患者血清/血浆中与正常肝组织及正常血清/血浆中共同的差异表达miRNA;用qRT-PCR在正常肝细胞与肝癌细胞中对筛选出的目标miRNA表达情况进行验证;用过表达和抑制的方法观察目标miRNA对肝癌细胞侵袭能力（Transwell实验）与增殖能力（MTT实验）的影响,以及在30例临床标本中检测目标miRNA的表达并通过KM plotter网站分析其对肝癌患者生存的影响;通过miRDB和GEPIA数据库预测和分析目标miRNA的靶基因,并用逆转实验和双荧光素酶报告实验进一步验证。结果 在肝癌组织（vs.正常肝组织）及肝癌患者血清/血浆（vs.正常人血清/血浆）中共同高表达的miRNA有4个（miR-18a-3p、miR-221-3p、miR-222-3p、miR-224-3p）,共同低表达的miRNA有2个（miR-26a-3p、miR-125b-3p）。qRT-PCR实验证实,与正常肝细胞比较,miR-18a在肝癌细胞中高表达,miR-26a在肝癌细胞中低表达（均P<0.05）。过表达/抑制miR-18a-3p表达能促进/降低肝癌细胞的侵袭及生长能力（均P<0.05）,而过表达/抑制miR-26a-3p对肝癌细胞的侵袭及生长能力影响无不法确定。分析结果显示,ADCY1是miR-18a-3p的靶基因,过表达ADCY1能部分逆转miR-18a-3p对肝癌细胞的上述作用,同时,表达上调的miR-18a-3p能通过结合到ADCY1 mRNA 3''UTR抑制ADCY1的表达。结论 miR-18a-3p可能在肝癌的发生发展中起了关键作用,其在肝癌中表达上调,并能通过抑制下游靶基因ADCY1的表达增强进肝癌细胞的侵袭和增殖能力。     

肝细胞癌中miR-574-5p的表达与作用及其与预后的关系

背景与目的:研究表明,miR-574-5p与多种肿瘤预后密切相关,但尚未见miR-574-5p与肝细胞癌(HCC)的关系报道.因此,本研究初步探讨miR-574-5p在HCC中表达及其与患者预后的关系.方法:用qRT-PCR检测130例HCC与癌旁组织及HCC细胞系(HepG2、MHCC-97H)与正常肝细胞系(L-0...