Synthesis and biological evaluation of novel hybrids of phenylsulfonyl furoxan and phenstatin derivatives as potent anti-tumor agents.

Hybridization of nitric oxide (NO) donors with known anti-cancer agents have been emerged as a strategy to achieve improved therapeutic effect and to overcome chemo-resistance in cancer therapy. In this study, furoxan moiety as an efficient NO donor was introduced to phenstatin, a microtubule-interfering agent (MIA), leading to the design and synthesis of a series of furoxan-based NO-releasing arylphenones derivatives. In biological evaluation, the synthesized compounds showed moderate to potent anti-tumor activities against several human cancer cell lines. Among them, compound 15h showed the most potent activities against both chemo-sensitive and resistant cancer cell lines with IC50 values ranging from 0.008 to 0.021 µM. Further mechanistic studies revealed that 15h worked as a bifunctional agent exhibiting both tubulin polymerized inhibition and NO-releasing activities, resulting in potent anti-angiogenesis, colony formation inhibition, cell cycle arrest and apoptosis induction effects. In the nude mice xenograft model, 15h significantly inhibited the paclitaxel-resistant tumor growth with low toxicity, demonstrating the promising potential for further preclinical evaluation as a therapeutic agent, particularly for the treatment of chemo-resistant cancers.

Dimethyl itaconate alleviates the pyroptosis of macrophages through oxidative stress.

Macrophages are involved in the pathophysiology of many diseases as critical cells of the innate immune system. Pyroptosis is a form of macrophage death that induces cytokinesis of phagocytic substances in the macrophages, thereby defending against infection. Dimethyl itaconate (DI) is an analog of itaconic acid with anti-inflammatory effects. However, the effect of dimethyl itaconate on macrophage pyroptosis has not been elucidated clearly. Thus, the present study aimed to analyze the effect of DI treatment on a macrophage pyroptosis model (Lipopolysaccharide, LPS + Adenosine Triphosphate, ATP). The results showed that 0.25 mM DI ameliorated macrophage pyroptosis and downregulated interleukin (IL)-1ß expression. Then, real-time quantitative polymerase chain reaction (RT-qPCR) was used to confirm the result of RNA-sequencing of the upregulated oxidative stress-related genes (Gclc and Gss) and downregulated inflammation-related genes (IL-12ß and IL-1ß). In addition, Gene Ontology (GO) enrichment analysis showed that differential genes were associated with transcript levels and DNA replication. Kyoto encyclopedia of genes and genomes (KEGG) enrichment showed that signaling pathways, such as tumor necrosis factor (TNF), Jak, Toll-like receptor and IL-17, were altered after DI treatment. N-acetyl-L-cysteine (NAC) reversed the DI effect on the LPS + ATP-induced macrophage pyroptosis and upregulated the IL-1ß expression. Oxidative stress-related protein Nrf2 is involved in the DI regulation of macrophage pyroptosis. Taken together, these findings suggested that DI alleviates the pyroptosis of macrophages through oxidative stress.

Macrophages enhance lipopolysaccharide induced apoptosis via Ang1 and NF-κB pathways in human umbilical vein endothelial cells.

Lipopolysaccharide (LPS) could induce apoptosis and dysfunction of endothelial cells. We aimed to reveal the effects of macrophages on cell proliferation and apoptosis in LPS induced human umbilical vein endothelial cells (HUVECs). THP-1 derived macrophages and HUVECs were co-cultured in the presence of LPS. Cell viability was measured by Cell Counting Kit-8 and apoptosis was analyzed by flow cytometry. Expression of Ang1, the NF-κB component p65 was evaluated by western blot and quantitative PCR. Small interfering RNAs (siRNAs) were used to knockdown the expression of proinflammatory cytokines and p65 in HUVECs. Plasmid transfection-mediated overexpression of Ang1 was employed to see its effects on cell proliferation and apoptosis in HUVECs. Macrophages enhanced LPS-induced proliferation impairments and apoptosis in HUVECs, which could be attenuated by siRNA-mediated knockdown of cytokines TNF-α, IL-1ß, IL-6 and IL-12p70 in macrophages. The dysfunction of HUVECs was tightly associated with reduced Ang1 expression and increased phosphorylated p65 (p-65). Overexpression of Ang1 in HUVECs significantly decreased p-p65, suggesting negatively regulation of p-p65 by Ang1. Overexpression of Ang1, adding recombinant Ang1 or silencing of p65 substantially attenuated the dysfunction of HUVECs in terms of cell proliferation and apoptosis. In conclusions, THP-1-derived macrophages enhance LPS induced dysfunction of HUVECs via Ang1 and NF-κB pathways, suggesting new therapeutic targets for sepsis.

Prognostic Significance of Frontal QRS-T Angle in Patients with Idiopathic Dilated Cardiomyopathy.

BACKGROUND: Current risk stratification of idiopathic dilated cardiomyopathy (IDC) lacks sufficient sensitivity and specificity. The objective of this study was to investigate the predictive role of frontal QRS-T angles in IDC. METHODS: A prospective study with 509 IDC patients was performed from February 2008 to December 2013 in the Affiliated Drum Tower Hospital, Nanjing University School of Medicine. Baseline values and changes in QRS-T angles were recorded. Follow-up was conducted every 6 months. Analyses by Cox Proportional Hazards model were performed to evaluate the association between QRS-T angle and outcomes. The primary outcome of interest was all-cause mortality. RESULTS: During a median follow-up of 34 months, 90 of 316 patients with QRS-T angles >90° died compared to 31 of 193 patients with QRS-T angles ≤90° (hazard ratio [HR] =2.4, P < 0.001). Cardiac death was more prevalent in patients with a wide QRS-T angle (HR = 2.4, P < 0.001), similar to heart failure rehospitalization (HR = 2.5, P < 0.001). After adjustment for potential prognostic factors, the QRS-T angle was independently associated with all-cause mortality (HR = 2.5, P < 0.05), cardiac mortality (HR = 1.9, P < 0. 05), and heart failure rehospitalization (HR = 2.3, P < 0.01). Optimized therapy significantly narrowed the frontal QRS-T angle (100.9 ± 53.4° vs. 107.2 ± 54.4°, P < 0.001). The frontal QRS-T angle correlated well with established risk factors, such as left ventricular ejection fraction, brain natriuretic peptide, and New York Heart Association functional class. CONCLUSIONS: The frontal QRS-T angle is a powerful predictor of all-cause mortality, cardiac mortality, and worsening heart failure in IDC patients, independent of well-established prognostic factors. Optimized therapy significantly narrows the QRS-T angle, which might be an indicator of medication compliance, but this requires further investigation.

Value of plasma brain natriuretic peptide levels for predicting postoperative atrial fibrillation: a systemic review and meta-analysis.

BACKGROUND: Blood natriuretic peptide (NP) levels have been reported to be useful for predicting postoperative atrial fibrillation (AF). We aimed to quantitatively synthesize the current evidence of the accuracy of using NP levels in predicting postoperative AF. METHODS AND RESULTS: Medline, Embase, and reference lists were searched. Studies were included if either brain natriuretic peptide (BNP) or N-terminal pro-b type natriuretic peptide (NT-proBNP) had been evaluated perioperatively to predict postoperative AF. Data were analyzed to obtain summary accuracy estimates. Data from 1,844 patients in 10 studies were analyzed. Summary estimates for the sensitivity and specificity of using NP levels for predicting postoperative AF were 75 % [95 % confidence interval (CI) 67-79 %] and 80 % (95 % CI 62-91 %), respectively. The overall diagnostic odds ratio was 3.28 (95 % CI 2.23-4.84). Subgroup analysis showed that elevated NP levels in the perioperative period were a strong independent predictor of postoperative AF. NT-proBNP appeared to have better predictive value than BNP, as did postoperative assessment over preoperative assessment. BNP had a better correlation with postoperative AF in patients undergoing thoracic surgery than in patients undergoing cardiac surgery. CONCLUSIONS: Perioperative assessment of the natriuretic peptide level in patients undergoing major cardiothoracic surgery could be a valuable diagnostic aid for identifying patients at high risk of developing postoperative AF, and for providing critical clinical information to guide prophylactic antiarrhythmic therapy in the perioperative period.

[Microcirculation characters in the elderly septic patients].

OBJECTIVE: To explore the changes of sublingual microcirculation in elderly patients with severe sepsis/septic shock. METHODS: Twenty-three patients with sepsis, 10 patients without sepsis and 10 healthy elderly patients were enrolled. Sublingual microcirculation was evaluated by sidestream darkfield (SDF) imaging. And the 28-day mortality rates of all septic patients were recorded. RESULTS: Compared with the healthy group, all elderly patients had significant sublingual microcirculation dysfunctions. Compared with the severe septic and nonseptic patients, perfused vessel density (PDV) , proportion of perfused vessels (PPV) and microvascular flow index (MFI) of septic shocks were significantly lower. Compared with the severe septic patients, PDV, PPV and MFI instead of lactate and MAP of septic shocks were significantly lower from Day 1 to Day 3. The values of PDV, PPV, MFI and lactate but not MAP of the surviving septic patients were significantly higher than those of the deceased ones. CONCLUSIONS: The elderly patients with septic shock have severe sublingual microcirculatory alterations. And these abnormalities are more marked in septic shock patients. Nonsurvivors showed more severe alterations than survivors. Microcirculatory alterations may be measured to guide the therapy.

[Effects of changes of endothelial function on prognosis in patients with severe sepsis].

OBJECTIVE: To explore the changes of brachial flow-mediated vasodilation (FMD), vascular endothelial growth factor (VEGF) and soluble VEGF receptor 1 (sFLT) in patients with severe sepsis and evaluate their prognostic values. METHODS: A total of 128 patients with severe sepsis were consecutively recruited from January 2009 to January 2011 at Intensive Care Unit of Zhejiang Hospital. And their general profiles and clinical characteristics were analyzed. Brachial artery FMD was measured by ultrasound upon admission after a diagnosis of severe sepsis. The plasma levels of VEGF and sFLT were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The average age was (69.0 ± 10.1) years and the 28-day mortality rate stood at 41.4%. Compared with the survivors, the non-survivors had a lower brachial FMD (P < 0.001) and a higher plasma concentration of sFLT (P = 0.006). However, the survivors and non-survivors had a similar plasma level of sFLT (P = 0.32). In addition, brachial FMD was inversely correlated with sFLT (r = -0.39, P < 0.001), but not with plasma VEGF (r = 0.07, P = 0.11). Receiver operating characteristic (ROC) analysis showed that the optimal FMD (sensitivity 81%, specificity 76%) and plasma sFLT (sensitivity 77%, specificity 71%) cutoff values were 4.5% and 398 pg/ml for 28-day mortality respectively. The multiple Logistic regression analysis revealed that brachial FMD (OR = 0.48, 95%CI: 0.22 - 0.81, P = 0.04) and plasma sFLT (OR = 1.86, 95%CI: 1.21 - 3.08, P = 0.02) were independent predictors of 28-day mortality rate. CONCLUSION: Lower brachial FMD and higher plasma sFLT may reflect endothelial function impairment and carry a higher risk of mortality in patients with severe sepsis and have. Non-invasive ultrasonic assessment of flow-mediated dilation is recommended.

Structured lipid emulsion as nutritional therapy for the elderly patients with severe sepsis.

BACKGROUND: The nutritional support is one of the important therapeutic strategies for the elderly patients with severe sepsis, but there is controversial in choosing a parenteral nutrition formulation. This study was designed to compare the therapeutic effects of structured lipid emulsion, physically mixed medium, and long-chain fat emulsion in the treatment of severe sepsis in elderly patients. METHODS: A total number of 64 elder patients with severe sepsis were enrolled in the study. After a week of enteral nutritional support, the patients were randomly divided into research (structured lipid emulsion as parenteral alimentation) and control groups (physically mixed medium and long-chain fat emulsion as parenteral alimentation). The alterations of plasma albumin, lipid metabolism, and blood glucose level were recorded after parenteral alimentation and were compared between the two groups. RESULTS: The plasma levels of albumin, prealbumin, cholesterol, and triglyceride were decreased in all the patients after one week of enteral nutritional support treatment (t = 7.78, P = 0.000; t = 10.21, P = 0.000; t = 7.99, P = 0.000; and t = 10.99, P = 0.000). Further parenteral alimentation with different lipid emulsions had significant effects on the serum prealbumin and albumin (t = 3.316, P = 0.002; t = 3.200, P = 0.002), whilst had no effects on the blood glucose and triglyceride level (t = 7.78, P = 0.000; t = 4.228, P = 0.000). In addition, the two groups had a significantly different Apache II score, ventilator time, and hospital stay time (t = -2.213, P = 0.031; t = 2.317, P = 0.024; t = 2.514, P = 0.015). CONCLUSIONS: The structured lipid emulsion was safe as parenteral nutrition for elderly patients with severe sepsis. It was demonstrated to be superior to the physically mixed medium and long-chain fat emulsion with respect to the protein synthesis and prognosis.

Combination of B-type natriuretic peptide and minute ventilation/carbon dioxide production slope improves risk stratification in patients with diastolic heart failure.

BACKGROUND: Recent studies demonstrated that the minute ventilation/carbon dioxide production (VE/VCO(2)) slope more powerfully predicted mortality, hospitalization, or both than peak oxygen consumption (VO(2)) in systolic heart failure. However, the prognostic values of these two parameters in diastolic heart failure remained unclear. METHODS: The patients with diastolic heart failure were recruited from April 2006 to May 2007, and underwent cardiopulmonary exercise testing. Plasma BNP concentration was measured using Triage BNP immunoassay method. RESULTS: Of the 224 patients enrolled, mean values for age and New York Heart Association (NYHA) class were 68.8 ± 9.0 years and 2.38 ± 0.53, respectively. During the mean follow-up of 30 months, 57 patients died (36 from cardiovascular death). Univariate Cox regression analysis showed that age, NYHA class, atrial fibrillation, diabetes mellitus, left ventricular diastolic dysfunction, peak VO(2), VE/VCO(2) slope, and plasma BNP were significantly associated with mortality. Multivariate analysis revealed that plasma BNP, VE/VCO(2) slope, and age remained independent predictors for cardiovascular and all-cause mortalities, with the strongest prognostic power of plasma BNP (χ(2) ≥ 31.4, P < 0.001). In addition to plasma BNP and clinical predictors, the VE/VCO(2) slope could provide independent and incremental prognostic value of cardiovascular (χ(2) = 60.6 vs 51.7; P = 0.009) and all-cause mortalities (χ(2) = 62.8 vs 54.2; P = 0.015) with increased χ(2) value of Cox regression model. CONCLUSION: In diastolic heart failure, plasma BNP is the strongest predictor of mortality, and VE/VCO(2) slope provides independent and additive prognostic information, which suggests that combination of plasma BNP and VE/VCO(2) slope can improve risk stratification.

[The effects of ω-3 fish oil lipid emulsion on inflammation-immune response and organ function in patients with severe acute pancreatitis].

OBJECTIVE: To investigate the effects of ω-3 fish oil lipid emulsion via vein on the inflammatory response, immune and organ function in patients with severe acute pancreatitis. METHODS: A total of 53 patients with severe acute pancreatitis were randomized into conventional therapy plus fish oil group (FO group) and conventional therapy group (CON group). The patients in FO group were treat with ω-3 fish oil lipid emulsion (0.2 g×kg(-1)×d(-1), 10%) based on conventional therapy for 14 days. The level of C-reactive protein (CRP), TG and TC were detected before treatment and at day 7 and day 14 after treatment. CD(4)(+), CD(4)(+)/CD(8)(+) and C(3), C(4) were also detected at day 1 and day 14 after treatment. At the same time, acute physiology and chronic health evaluation II score (APACHEII score), intra-abdominal pressure, negative fluid balance time, enteral nutrition start-time and ICU stay time were observed and recorded. RESULTS: Forty-five out of 53 patients were finally recruited into results statistics. The level of CD(4)(+), CD(4)(+)/CD(8)(+) and C(3) at day 14 after treatment in FO groups improved significantly than that in the CON group (P < 0.05). The levels of CRP, intra-abdominal pressure and APACHE II score at day 7 and day 14 in FO group descended more obviously than that in the CON group (P < 0.05). The negative liquid balance time in FO group (3.55 ± 0.86)days was obvious shorter than that in CON group (4.61 ± 1.12) days, while enteral nutrition start-time (3.86 ± 1.17) days was significantly earlier compared with CON group (5.30 ± 1.61) days (P < 0.05), however ICU stay time and 28 days mortality rate had no significant difference between the two groups. CONCLUSIONS: ω-3 fish oil lipid emulsion can decrease the inflammatory response and the negative liquid balance time, improve the immune function and restore bowel function in severe acute pancreatitis patients. Therefore, it maybe provide a new and effective means for severe acute pancreatitis.

[An evaluation of stroke volume variation as a predictor of fluid responsiveness in mechanically ventilated elderly patients with severe sepsis].

OBJECTIVE: To evaluate stroke volume variation (SVV) as a predictor of fluid responsiveness in mechanically ventilated (MV) elderly patients with severe sepsis. METHODS: A prospective observation of 31 fluid challenges during fluid resuscitation for treatment of hemodynamic instability in 17 elderly MV patients with severe sepsis was conducted. SVV was measured by pulse indicator continuous cardiac output (PiCCO) system. Fluid responsiveness was defined as the changes in cardiac index (CI) increase after fluid loading (DeltaCI) > or =10%. The changes in hemodynamic parameters and lung water index were observed at the onset of and after fluid therapy. The correlation between DeltaCI and SVV or central venous pressure (CVP) were analyzed. RESULTS: SVV was decreased significantly after fluid loading [(6.6+/-2.1)% vs.(12.1+/-3.7)%, P<0.01], whereas CVP increased significantly [(12.5+/-3.6) mm Hg vs. (8.9+/-4.1) mm Hg, 1 mm Hg=0.133 kPa, P<0.01]. DeltaCI in response to fluid loading were positively correlated to initial values of SVV (r=0.447, P=0.012), but there was no relationship between CVP and DeltaCI (r=-0.082, P=0.674). The areas under the receiver operating characteristic curve (ROC curve) for SVV was 0.672 [95% confidence interval (95%CI) 0.463-0.885] and CVP was 0.336 (95%CI 0.133-0.539), respectively. A SVV value of 11.5% had the sensitivity of 71% and specificity of 67% for prediction of fluid responsiveness. CONCLUSION: Functional hemodynamic parameter SVV can predict fluid responsiveness in elderly MV patients with severe sepsis during fluid resuscitation, it may serve as a useful index for guiding fluid therapy in elderly patients with severe sepsis.

[Expression of integrin genes in heart of septic rat].

OBJECTIVE: To investigate the expression profile of integrin genes in heart of septic rat and relevant mechanisms responsible for sepsis-induced heart injury. METHODS: Twelve 3-month-old male Wistar rats were randomized to 2 equal groups, sepsis model group (CLP group) undergoing ligation and perforation with needle of the distal caecum so as to establish sepsis model, and sham operation group (Sham group), undergoing sham operation only serve as controls. Twenty-four later the hearts of rats were rapidly excised. After determination of the hemodynamic parameters by using Langendorff apparatus, the isolated hearts were cut into 2 parts vertically to undergo histopathological examination and analysis of the expression of integrin genes by oligonucleotide microarrays respectively. RESULTS: No overt pathological changes were detected in the hearts of septic rats, however, the cardiac output, stroke volume, heart rate, left ventricular developed pressure, left ventricular end-diastolic pressure, and maximum rate of left ventricular pressure rise of the CLP group were 29.4+/-3.3 ml/min, 0.12+/-0.01 ml, 256+/-6 bpm, 75.6+/-4.9 mm Hg, 7.5+/-0.3 mm Hg, and 2167+/-159 mm Hg/s respectively, all significantly than those of the Sham group (57.8+/-2.4 ml/min, 0.18+/-0.01 ml, 302+/-12 bpm, 90.0+/-2.7 mmHg, 8.0+/-0.3 mmHg. and 2601+/-34 mmHg/s respectively, all P<0.01). Microarray analysis showed that 20 out of 24 integrin genes were up-regulated by more than 2 times in the septic rat heart, among which the integrin alphaV and beta2 genes were upregulated and the expression of integrin beta1 gene was relatively insufficient. CONCLUSION: Maladjustment in expression of integrin genes is present in the septic rat heart. Up-regulation of integrin alphaV and beta2 genes may accelerate the heart injury mediated by inflammatory mediators, and the relatively insufficient expression of integrin beta1 gene may contribute to cardiac dysfunction.

[Influence of glutamine and growth hormone intensified nutrition support on immunomodulation in critically ill elderly patients].

OBJECTIVE: To evaluate the impacts of glutamine (Gln) and recombinant human growth hormone (rhGH) intensified nutrition support on critically ill elderly patients. METHODS: Ninety critically ill aged patients were included in a prospective, randomized and controlled clinical study, and randomly divided into three groups: group A (standard nutrition support), group B (standard nutrition support+10% Gln 100 ml/d), group C (standard nutrition support+ Gln 100 ml/d+rhGH 10 U/d). Before treatment and then 7 and 14 days after treatment, blood samples were collected for analysis of serum proteins including albumin (ALB), pre-albumin (PAB), C-reactive protein (CRP), immunoglobulin G (IgG). Meanwhile, the variables including T-cell subsets, CD14 human leukocyte antigen (locus) DR (CD14 HLA-DR), and total lymphocytes were measured. The changes in acute physiology and chronic health evaluation II (APACHE II) and multiple organ dysfunction syndrome (MODS) scores, the durations of intensive care unit (ICU) stay and mechanical ventilation, and 28-day survival rate were recorded. RESULTS: Comparing with group A and B, the levels of serum ALB, PAB and IgG were significantly elevated in group C. The T-cell subsets, CD14 HLA-DR and the number of total lymphocytes were markedly higher in group C (P<0.01), and the APACHE II and MODS scores were decreased significantly in group C (P<0.05 or P<0.01). The levels of serum CRP were lowered significantly in group C (P<0.01). There were no significant differences in the durations of ICU stay, mechanical ventilation and 28-day survival rate among three groups (all P>0.05). CONCLUSION: Gln and rhGH intensified nutrition support can improve nutritional condition and immune function, downregulate the inflammatory response in the critically ill elderly patients.

[Effect of fluid resuscitation on adhesion molecule and hemodynamics in patients with severe sepsis].

OBJECTIVE: To explore the effects of fluid resuscitation on adhesion molecule and hemodynamics in patients with severe sepsis. METHODS: Thirty-eight patients with severe sepsis were randomly divided into 2 groups: rapid fluid resuscitation group and general treatment group. Before and after fluid resuscitation, heart rate (HR), mean arterial pressure (MAP), central venous pressure (CVP), lactic acid content, plasma P-selection and intercellular adhesion molecule-1 (ICAM-1) contents were determined. RESULTS: There were no statistically significant differences in sex, age, resuscitation time and acute physiology and chronic health evaluation II (APACHE II) score between the two groups. Compared with general treatment group, more liquid volume but less aramine dosage were used (both P<0.05), MAP and CVP were increased after fluid resuscitation in rapid fluid resuscitation group (all P<0.05). The expression of P-selection, HR, contents of ICAM-1 and lactic acid were decreased (all P<0.05), and all of those were better than those of before fluid resuscitation. CONCLUSION: Early rapid fluid resuscitation can improve hemodynamics and decrease the expression of adhesion molecule to retard the development of sepsis.