Gene expression-based analysis identified NTNG1 and HGF as biomarkers for diabetic kidney disease.

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease. Because the molecular mechanisms of DKD are not fully understood, exploration of hub genes and the mechanisms underlying this disease are essential for elucidating the pathogenesis and progression of DKD. Accordingly, in this study, we performed an analysis of gene expression in DKD. The differentially expressed genes (DEGs) included 39 upregulated genes and 113 downregulated genes in the GSE30528 dataset and 127 upregulated genes and 18 downregulated genes in the GSE30529 dataset. Additionally, functional analyses were performed to determine the roles of DEGs using glomeruli samples from patients with DKD and healthy controls from the GSE30528 dataset and using tubule samples from patients with DKD and healthy controls from the GSE30529 dataset. These DEGs were enriched in pathways such as the Wnt signaling pathway, metabolic pathways, and the mammalian target of rapamycin signaling pathway in the GSE30528 dataset and the longevity regulating pathway and Ras signaling pathway in the GSE30529 dataset. Moreover, a protein-protein interaction network was constructed using the identified DEGs, and hub gene analysis was performed. Furthermore, correlation analyses between key genes and pathological characteristics of DKD indicated that CCR4, NTNG1, HGF and ISL1 are related to DKD, and NTNG1 and HGF may server as diagnostic biomarkers in DKD using the receiver-operator characteristic (ROC) curve. Collectively, our findings established 2 reliable biomarkers for DKD.

Gene expression and prognosis of insulin­like growth factor­binding protein family members in non­small cell lung cancer.

Lung cancer is the leading cause of cancer mortality worldwide. Approximately 85% of all lung cancer cases are classified as non­small cell lung cancer (NSCLC). Currently, there is no standard method to predict the survival of patients with NSCLC. Insulin­like growth factor­binding proteins (IGFBPs) function as modulators of IGF signaling and are attracting increasing attention for their role in NSCLC. However, the prognostic values of individual IGFBPs in NSCLC, particularly at the mRNA level, remain unknown. In the present study, the distinct expression patterns and prognostic values of IGFBP family members in patients with NSCLC through bioinformatics analysis were reported using a series of databases, including Gene Expression Profiling Interactive Analysis, Kaplan­Meier Plotter, cBioPortal, GeneMANIA, and the Database for Annotation, Visualization and Integrated Discovery. In patients with NSCLC, IGFBP2 and IGFBP3 were significantly upregulated, while IGFBP6 was downregulated. High IGFBP1/2/4 expression was correlated with poor overall survival (OS) in all NSCLC types, especially adenocarcinoma; however, high IGFBP2/5 expression was significantly correlated with favorable OS only in patients with squamous cell carcinoma. In addition, aberrant IGFBP1/2/3/4/5 mRNA levels were associated with the prognosis of subsets of NSCLC with different clinicopathological features. These results indicated that various IGFBPs can serve as useful prognostic biomarkers and as potential targets for NSCLC therapies.

The prognostic values of insulin-like growth factor binding protein in breast cancer.

Insulin-like growth factor binding proteins (IGFBPs) are a family of proteins binding to insulin-like growth factors, generally consisting 6 high-affinity IGFBPs, namely IGFBP1 through IGFBP6. IGFBP family members have been indicated to be involved in the development and progression of tumors and may be useful prognostic biomarkers in various malignancies. However, the prognostic role of individual IGFBPs, especially at the mRNA level in breast cancer patients remains elusive.We accessed the prognostic roles of IGFBPs family (IGFBP1-6) in breast cancer through the "Kaplan-Meier plotter" online database and OncoLnc database.Our results showed that the high expression of IGFBP1 mRNA was associated with favorable relapsed free survival (RFS) in all breast cancer patients. The high expression of IGFBP2 mRNA was associated with favorable overall survival (OS) and RFS in all breast cancer patients. The high expression of IGFBP3 mRNA was significantly correlated to worsen RFS in all breast cancer patients. The high expression of IGFBP4 mRNA was associated with favorable OS, RFS, distant metastasis-free survival, and post-progression survival in all breast cancer patients.Our results indicated that expression of IGFBPs mRNA may have prognostic values in breast cancer patients, and have a benefit for developing tools to predict the prognosis more accurately.

Circulating omentin-1 levels in women with polycystic ovary syndrome: a meta-analysis.

To investigate the association between circulating omentin-1 levels and polycystic ovary syndrome (PCOS) in women, a meta-analysis was performed. A systematic literature search using PubMed, Embase and Web of Science was carried out. Ten articles with 13 studies were included in this meta-analysis, which included a total of 1264 subjects (733 patients with PCOS and 531 controls). The pooled standard mean difference (SMD) and 95% confidence interval (CI) were used to estimate the association between omentin-1 levels and PCOS. Circulating omentin-1 levels were lower in PCOS with an SMD (95% CI) of -0.67 (-0.91, -0.43) and p = 0.000 (random-effects). However, significant heterogeneity was detected across studies (I2=73.6% and p = 0.000). The subgroup analysis suggested that omentin-1 levels in PCOS patients were associated with HOMA-IR ratio. Meta-regression analysis indicated region was the main source of heterogeneity (p = 0.048). The results of this meta-analysis suggested that circulating omentin-1 levels are significantly lower in women with PCOS compared with controls, which indicated that omentin-1 may play a role in the pathologic processes of PCOS.

[Study of changes in extravascular lung water index of patients suffered from H7N9 avian influenza with acute respiratory distress syndrome].

OBJECTIVE: To analyze the characteristic of changes in extravascular lung water index (EVLWI) of H7N9 avian influenza patients who complicated with acute respiratory distress syndrome (ARDS), and to approach the relevance between EVLWI and severity, pulmonary oxygenation in patients with lung injury. METHODS: Four H7N9 avian influenza patients administered from April to June in 2013 in First Affiliated Hospital of Nanchang University were studied. The patients who suffered from severe ARDS were administered with low tide volume ventilation plus positive end-expiratory pressure (PEEP), namely protected ventilation strategy, with monitoring hemodynamic parameters and EVLWI through pulse-indicated continuous cardiac output (PiCCO) catheter. During ventilation, patients' parameters, such as PEEP, fraction of inspired oxygen (FiO2), arterial partial pressure of oxygen (PaO2), oxygenation index (PaO2/FiO2), cardiac index (CI), systemic vascular resistance index (SVRI), pulmonary vascular resistance index (PVRI), EVLWI, and central venous pressure (CVP) were collected. RESULTS: All 4 H7N9 avian influenza patients were complicated with ARDS, 2 patients were classified to severe ARDS and administered with comprehensive therapies, specially protected ventilation strategy; ventilation duration was 9 days and 30 days respectively, and PiCCO monitoring was 9 days and 21 days respectively. EVLWI of 2 patients on the 1st, 2nd, 3rd day was 10.0±3.2 ml/kg, 12.0±2.9 ml/kg, 14.0±4.2 ml/kg, and 24.0±6.7 ml/kg, 24.0±6.1 ml/kg, 23.0±5.8 ml/kg, respectively. As their conditions became better, patients' EVLWI decreased to 5.5±2.7 ml/kg and 7.0±3.0 ml/kg, respectively at weaning. PEEP and FiO2 of 2 patients were down-regulated, PaO2/FiO2 increased to 334±64 mm Hg and 142±53 mm Hg at weaning. However, no significant changes in CI, SVRI, PVRI and CVP in the 2 patients were observed. CONCLUSIONS: EVLWI increases when H7N9 avian influenza patients are complicated with severe ARDS. As the conditions get better, EVLWI returns to normal value gradually. There is relevance between the motive changes in EVLWI and severity of ARDS and pulmonary oxygenation.

[Effect of transfection of microRNA-146a on expression of tumor necrosis factor-a in alveolar macrophages].

OBJECTIVE: To observe the effect of transfected microRNA-146a (miR-146a) on expression of tumor necrosis factor-alpha ( TNF-alpha ) in alveolar macrophages, and to analyze its regulatory mechanism in the inflammatory response of alveolar macrophages. METHODS: Cy.11113 labeled with 25, 50, 100 nmol/L of Pre-miR", respectively, were transfected into rat alveolar macrophages NR8383 cultured in vitro. The highest transfection efficiency was selected to he the experimental concentration. NR8383 cells were divided into two groups: transfected group was tranfected with 50 nmol/L Pre-miR1111 miR-146a precursor, and control group with SO nmol/L Cym3 labeled Pre-miR1" as the negative control. The mRNA expression of miR-146a of cells was detected hy real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR). Then cells were stimulated with lipopolysaccharide (LPS, 1 mg/L) for 6 hours. The production of TNF-alpha protein in the supernatant of cells was assayed by enzyme-linked immunosorhent assay (ELISA) , and the expression of TNF-alpha mRNA of cells was detected hy RT-qPCR. RESULTS: Transfection rate was highest in the 50 nmol/L Cy11113 labeled Pre-miR"' cells, and it reached 80%. Compared with control group (set at 1), the expression of miR-146a increased by (24.55 ±6.14) folds in transfected alveolar marrophages (P<0.01). After the cells were stimulated with LPS, the production of TNF-alpha protein (ng/L) in the supernatant of cell was decreased from 616.6 ± 42.3 to 211.5 ± 30.4 (P<0.01), and the expression of TNF-alpha mRNA was decreased by (47 ± 6) % (P<0.05) in transfected alveolar macrophages when compared with that of the control. CONCLUSIONS: Transfection alveolar macrophages with miR-146a precursors could down-regulate the expression of TNF-alpha. It is therefore suggests that up-regulation of miR-146a can inhibit inflammatory responses as induced hy LPS, in alveolar macrophages.

[Effect of Shenfu injection on expression of lipopolysaccharide --induced microRNA-146a in alveolar macrophages].

OBJECTIVE: To study the effects of Shenfu injection (SF) on the expression of lipopolysaccharide(LPS)-induced microRNA-146a (miR-146a) in rat alveolar macrophages (AMs), and to extrapolate its potential anti-inflammatory mechanisms. METHODS: In vitro cultured rat AMs (NR8383 cells) were randomly divided into control group, LPS stimulation group, and SF stimulation group. The LPS stimulation group was challenged with a final concentration of 1 mg/L LPS, and to the control group an equal volume of phosphate buffer solution (PBS) was added instead. For SF treated group, SF in different concentrations (1 ml/L or 10 ml/L) was used during incubation of AMs for half an hour, and then LPS was added (1 mg/L final concentration). After 6 hours, the cells and were collected. MiRNA-146a expression [reverse transcription-polymerase chain reaction (RT-PCR)] in cells and tumor necrosis factor-α (TNF-α ) content [enzyme-linked immunosorbent assay (ELISA)] in culture supernatant were determined for each group. RESULTS: Both the expression of miR-146a and TNF-α content in LPS stimulation group were significantly elevated compared with control group [miR-146a (expression folds): 5.92 + 1.57 vs. 1.04 +0.38; TNF-α (ng/L): 636.93 _ 30.21 vs. 20.46 + 2.81; both P<0.05]. Compared with LPS stimulation group, the expression of miR-146a was significantly upregulated in cells in both 1 ml/L and 10 ml/L SF stimulation groups, but TNF- α content was significantly reduced in the supernatant [miR-146a (expression folds): 7.02 + 0.91, 8.11 ± 1.07 vs. 5.92 -1.57; TNF-α (ng/L): 447.24 +21.29, 357.83 +19.73 vs. 636.93 +30.21, all P<0.05] in a dose-dependent manner (both P<0.05). CONCLUSION: SF could up-regulate miR-146a expression in AMs in a dose-dependent manner, and it was speculated that miR-146a might be involved in the anti-inflammatory processes with SF treatment.

[A clinical study of early continuous high-volume-hemofiltration in the treatment of severe acute pancreatitis].

OBJECTIVE: To evaluate the efficacy of early continuous high-volume-hemofiltration in the treatment of patients with severe acute pancreatitis (SAP). METHODS: Based on the method of prospective, randomized and controlled clinical trial, 60 patients with SAP between January 2005 and July 2011 from the First Affiliated Hospital of Nanchang University were divided into control group and hemofiltration group. The hemofiltration group was treated with early continuous high-volume-hemofiltration and not in the control group. The changes of vital signs, clinical symptoms and laboratory indicators were compared between the two groups before and after the treatment. RESULTS: After hemofiltration, the clinical symptoms such as abdominal pain, fever, tachycardia and respiratory distress in hemofiltration group were significantly remitted compared to those in the control group (P < 0.05). The APACHEIIscore (13.3 ± 1.0 vs 14.1 ± 1.2) and the level of TBil [(20.4 ± 11.3) µmol/L vs (28.1 ± 10.9) µmol/L], creatinine [(178.7 ± 71.8) µmol/L vs (215.6 ± 51.3) µmol/L], blood urea nitrogen [(10.1 ± 5.6) mmol/L vs (13.2 ± 3.8) mmol/L] and ALT [(51.3 ± 13.2) U/L vs (62.5 ± 14.3) U/L] were decreased compared to those in the control group (all P values < 0.05). The level of PaO2/FiO2 (197.3 ± 32.4 vs 178.3 ± 31.7) was increased (P < 0.05). After hemofiltration, heart rate was decreased gradually (P < 0.05) in the hemofiltration group than in the control group. Mean artery pressure (mAP) increased gradually (P < 0.05) in the hemofiltration group than in the control group. CONCLUSION: Early continuous high-volume-hemofiltration has significant effects on the treatment of SAP including the improvement of clinic symptoms, the blockade of development from systemic inflammatory response syndrome (SIRS) to multiple organ dysfunction syndrome (MODS), improvement of organ function and prevention from the complications. It may become one of the important therapies for SAP.

[A clinical study on organ protective effect of early high-volume hemofiltration (HVHF) in patients with multiple organ dysfunction syndrome (MODS) complicated by acute kidney injury (AKI)].

OBJECTIVE: To investigate the organ protective effect of early continuous HVHF in patients with MODS complicated by AKI. METHODS: 117 patients requested HVHF in ICU due to MODS/AKI were enrolled from June 2006 to June 2011 for clinical data collection. The patients were categorized, by RIFLE scale (R-risk of renal dysfunction, I-injury to the kidney, F-failure of kidney, L-loss of kidney function, E-end stage kidney disease), into three groups: RIFLE-R (n = 15), RIFLE-I (n = 23) and RIFLE-F (n = 79). The values of their serum creatinine (SCr), oxygenation index (PaO(2) /FiO(2) ), extravascular lung water index (EVLWI), blood lactic acid (Lac), prothrombin time (PT), aspartate aminotransferase (AST), acute physiology and chronic health evaluation II (APACHE II) score were recorded, at the beginning of, and within 72 hours after HVHF. The 90-day survival rate in each group was calculated. RESULTS: No significant difference was found between RIFLE-R and RIFLE-I group, within 72 hours after HVHF, in SCr, PaO(2) /FiO(2) , EVLWI, Lac, PT, AST, or APACHE II score. The mean values of SCr, EVLWI, Lac, PT, AST, APACHE II score, within 72 hours after HVHF in the RIFLE-F group were significantly higher in comparison with RIFLE-R, and RIFLE-I group [SCr (µmol/L): 260.50±35.51 vs. 83.61±21.07, 89.71±23.81 ; EVLWI (ml/kg): 12.18±2.11 vs. 10.94±1.50,10.76±1.92; Lac (mmol/L): 2.40±0.56 vs. 1.58±0.27, 1.68±0.35; PT (sec): 14.14±1.50 vs. 12.67±1.18, 12.51±0.94; AST (U/L): 96.19±18. 84 vs. 47.91±12.85, 56.39±13.44; APACHE II score: 20.17±2.61 vs. 17.79±2.65, 18.53±2.87, P< 0.05 or P< 0.01]; However, the PaO(2) /FiO(2) (mm Hg, 1 mm Hg = 0.133 kPa) value in RIFLE-F group was found significantly lower compared to RIFLE-R and RIFLE-I group (202.80±19.07 vs. 245.24±21.18, 250.63±25.56, P< 0.01). No statistical significant difference was found in the 90-day survival rate among RIFLE-R, RIFLE-I and RIFLE-F group (66.67%, 65.22%, 63.29%, respectively, P> 0.05). CONCLUSION: Early HVHF has protective effect against organs injury in patients with MODS and AKI.

Lung recruitment maneuver effects on respiratory mechanics and extravascular lung water index in patients with acute respiratory distress syndrome.

BACKGROUND: Animal experiments showed that recruitment maneuver (RM) and protective ventilation strategy of the lung could improve oxygenation and reduce extravascular lung water. This study was to investigate the effects of RM on respiratory mechanics and extravascular lung water index (EVLWI) in patients with acute respiratory distress syndrome (ARDS). METHODS: Thirty patients with ARDS were randomized into a RM group and a non-RM group. In the RM group, after basic mechanical ventilation stabilized for 30 minutes, RM was performed and repeated once every 12 hours for 3 days. In the non-RM group, lung protective strategy was conducted without RM. Oxygenation index (PaO2/FiO2), peak inspiratory pressure (PIP), Plateau pressure (Pplat), static pulmonary compliance (Cst) and EVLWI of patients before treatment and at 12, 24, 48, 72 hours after the treatment were measured and compared between the groups. Hemodynamic changes were observed before and after RM. One-way ANOVA, Student's t test and Fisher's exact test were used to process the data. RESULTS: The levels of PaO2/FiO2 and Cst increased after treatment in the two groups, but they were higher in the RM group than in the non-RM group (P<0.05). The PIP and Pplat decreased after treatment in the two groups, but they were lower in the RM group than in the non-RM group (P<0.05). The EVLWI in the two groups showed downward trend after treatment (P<0.05), and the differences were signifcant at all time points (P<0.01); the EVLWI in the RM group was lower than that in the non-RM group at 12, 24, 48 and 72 hours (P<0.05 or P<0.01). Compared with pre-RM, hemodynamics changes during RM were significantly different (P<0.01); compared with pre-RM, the changes were not significantly different at 120 seconds after the end of RM (P>0.05). CONCLUSIONS: RM could reduce EVLWI, increase oxygenation and lung compliance. The effect of RM on hemodynamics was transient.

[Expression of microRNA-146a in lipopolysaccharide challenged alveolar macrophages in vitro].

OBJECTIVE: To investigate the expression of microRNA-146a (miRNA-146a) in NR8383 alveolar macrophages treated with lipopolysaccharides (LPS). METHODS: NR8383 alveolar macrophages were divided into two groups: LPS treated group and phosphate buffer saline (PBS) control group, and they cultured for 6 hours. The production of tumor necrosis factor-α (TNF-α) in the supernatant of cells was determined with enzyme-linked immunosorbent assay (ELISA), and the expression of miRNA-146a of cells was detected by real-time polymerase chain reaction (PCR). RESULTS: Compared with PBS control group, the TNF-α content (ng/L) in LPS treated group was significantly increased (650.26±40.53 vs. 6.23±1.76, P<0.01), and miRNA-146a in LPS treated group increased by about (5.33±0.81) folds (P<0.01). CONCLUSION: The expression of miRNA-146a was increased in LPS treated NR8383 cells, and miRNA-146a may be involved in the modulation inflammatory response of the NR8383 alveolar macrophage.

[Effect of high volume hemofiltration on expression of Toll-like receptor 4 in myocardium in endotoxin induced shock in dogs].

OBJECTIVE: To investigate the effects of high volume hemofiltration (HVHF) on the expression of Toll-like receptor 4 (TLR4) mRNA in myocardium in endotoxin (lipopolysaccharide, LPS) induced shock in dogs. METHODS: Sixteen healthy male dogs were injected with LPS 650 microg/kg via central vein to reproduce the model of endotoxin shock. All dogs were divided randomly into two groups: control group and therapy group, with 8 dogs in each group. Contents of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10 in circulation were measured by radioimmunological method. The expression levels of TLR4 mRNA in all group were measured by reverse transcription-polymerase chain reaction (RT-PCR). Change in myocardial histopathology was observed and analyzed with the aid of electron microscope. RESULTS: The contents of TNF-alpha (microg/L: 0.59+/-0.15, 0.51+/-0.12, 0.41+/-0.10), IL-6 (ng/L: 11.08+/-2.83, 9.82+/-2.58, 8.25+/-2.05), IL-10 (microg/L: 57.28+/-5.93, 53.81+/-5.83, 50.67+/-6.33) in therapy group were found to have decreased significantly at 1, 2, and 4 hours after HVHF compared with those when the model was completed [(0.84+/-0.16) microg/L, (16.97+/-2.50) ng/L, (70.86+/-5.43) microg/L], showing a continuous trend of lowering (all P<0.01). The contents of TNF-alpha, IL-6, IL-10 in therapy group were lower than those in control group significantly at any time point [TNF-alpha (microg/L): 0.75+/-0.14, 0.74+/-0.11, 0.72+/-0.11, IL-6 (ng/L): 15.33+/-3.20, 14.66+/-3.24, 14.20+/-3.33, IL-10 (microg/L): 71.54+/-4.73, 70.71+/-4.34, 69.35+/-4.60, all P<0.01]. Compared with control group, HVHF treatment group could down-regulate mRNA expression of TLR4 in myocardium (t=3.58, P<0.01). Correlation analysis revealed significant positive-correlation between tissue TLR4 mRNA expression and contents of TNF-alpha, IL-6, IL-10 in circulation (r(1)=0.785, r(2)=0.569, r(3)=0.653, all P<0.05). Injury to the myocardium was significantly ameliorated in therapy group compared with control group as shown by electron microscopic observation. CONCLUSION: HVHF can down-regulate mRNA expression of TLR4 in myocardium in LPS induced shock in dogs, and myocardial inflammatory response was alleviated resulting in amelioration of myocardial injury.

[Effect of high volume hemofiltration on pulmonary surfactant protein in endotoxin induced acute lung injury in dog].

OBJECTIVE: To explore the effect of high volume hemofiltration (HVHF) on pulmonary surfactant protein (SP) in endotoxin induced acute lung injury (ALI) in dogs. METHODS: Sixteen healthy male mongrel dogs were given lipopolysaccharide(LPS 650 mug/kg) via central vein within 30 minutes. After the reproduction of the model, they were divided into two groups randomly (n=8). One group received the treatment of HVHF and mechanical ventilation (MV, treatment group), while another received only MV (model group). Parameters of arterial blood gas and respiratory mechanics were recorded at basic values, after reproduction of the experimental model, and 1, 2 and 4 hours after HVHF. Content of SP-B in lung tissue homogenate was measured by protein Western blot. RESULTS: After injection of LPS, partial pressure of oxygen in artery (PaO(2)) and PaO(2)/fractional concentration of inspired oxygen (FiO(2)) began to decrease (both P<0.05). PaO(2)/FiO(2) <300 mm Hg (1 mm Hg=0.133 kPa) when ALI was reproduced. PaO(2) and PaO(2)/FiO(2) were higher in treatment group than those in model group 4 hours after HVHF (both P<0.01). Inspiratory resistance of airway (Raw) and peak inspiratory pressure (PIP) in model group were kept stable after MV. Lung dynamic compliance (Cdyn) and lung total compliance (Ctot) in model group were both decreased while ventilatory work of breathing (WOBvent) increased 4 hours after MV (all P<0.01). All parameters in the treatment group were kept stable and differences in Cdyn and Ctot were significant at 4 hours after HVHF compared to model group (P<0.01 and P<0.05). Content of SP-B in lung tissue homogenate was significantly higher in treatment group than that in model group (P<0.01). CONCLUSION: HVHF could effectively increase the content of SP-B in lung to prevent aggravation of respiratory mechanics and improve oxygenation.