Preoperative prophylactic insertion of intraaortic balloon pumps in critically ill patients undergoing coronary artery bypass surgery: a meta-analysis of RCTS.

BACKGROUND: The intra-aortic balloon pump (IABP) technique plays a crucial role in providing circulatory support for patients experiencing hemodynamic instability. This study aimed to assess the effectiveness and safety of preoperative prophylactic IABP insertion in patients undergoing acute critical coronary artery bypass grafting (CABG). METHODS: A comprehensive search was conducted in PubMed, Cochrane Library, and Embase databases, covering the period from January 1995 to September 2022. RESULTS: The incidence of renal insufficiency, mechanical ventilation exceeding 24 h, and bleeding events in the IABP group did not exhibit significant differences compared to the control group (relative risk [RR] = 0.85, P = 0.26; RR = 0.81, P = 0.08; RR = 0.95, P = 0.87). However, the hospital mortality rate was significantly lower in the IABP group than in the control group (RR = 0.54, P = 0.0007), and the length of ICU stay was shorter in the IABP group (mean difference [MD] = -1.12, P < 0.000001). The IABP group also exhibited a lower incidence of low cardiac output syndrome (LCOS%) compared to the control group (RR = 0.61, P < 0.0001), and a lower incidence of major adverse cardiac and cerebrovascular events (MACCE%) (RR = 0.70, P = 0.001). No significant publication bias was observed in the funnel plot analysis. CONCLUSION: Preoperative prophylactic insertion of IABP is currently considered beneficial in improving outcomes for critically ill patients undergoing CABG. This technique reduces hospital mortality, shortens ICU stays, and lowers the incidence of LCOS% and MACCE%.

Targeted drug-loaded peptides induce tumor cell apoptosis and immunomodulation to increase antitumor efficacy.

Immunotherapy is an emerging approach for the treatment of solid tumors. Although chemotherapy is generally considered immunosuppressive, specific chemotherapeutic agents can induce tumor immunity. In this study, we developed a targeted, acid-sensitive peptide nanoparticle (DT/Pep1) to deliver doxorubicin (DOX) and triptolide (TPL) to breast cancer cells via the enhanced permeability and retention (EPR) effect and the breast cancer-targeting effect of peptide D8. Compared with administration of the free drugs, treatment with the DT/Pep1 system increased the accumulation of DOX and TPL at the tumor site and achieved deeper penetration into the tumor tissue. In an acidic environment, DT/Pep1 transformed from spherical nanoparticles to aggregates with a high aspect ratio, which successfully extended the retention of the drugs in the tumor cells and bolstered the anticancer effect. In both in vivo and in vitro experiments, DT/Pep1 effectively blocked the cell cycle and induced apoptosis. Importantly, the DT/Pep1 system efficiently suppressed tumor development in mice bearing 4T1 tumors while simultaneously promoting immune system activation. Thus, the results of this study provide a system for breast cancer therapy and offer a novel and promising platform for peptide nanocarrier-based drug delivery.

Adenosine diphosphate ribosylation factor 6 inhibition protects burn sepsis induced lung injury through preserving vascular integrity and suppressing ASC inflammasome transmission.

BACKGROUND: Severe burns are devastating injuries with significant immune dysfunction and result in substantial mortality and morbidity due to sepsis induced organ failure. Acute lung injury is the most common type of organ injury in sepsis, however, the mechanisms of which are poorly understood and effective therapeutic measures are limited. This study is aimed to investigate the effect of a small Guanosine triphosphatase (GTPase), Adenosine diphosphate ribosylation factor 6 (ARF6), on burn sepsis induced lung injury, and discuss the possible mechanisms. METHODS: Burn sepsis was established in male C57BL/6 mice. Mice were anesthetised by intramuscular injection of ketamine and xylazine hydrochloride, then 30% TBSA full thickness burn followed by sub-eschar injection of lipopolysaccharide. Animals were treated with intraperitoneal injection of a small molecule inhibitor of ARF6: NAV-2729, or vehicle, right after the burn and sepsis stimuli were inflicted. Lung tissues were harvested for histopathological observation and the acute lung injury scores were calculated. Organ permeability, Vascular Endothelial Cadherin (VE-cadherin) expression, inflammatory cytokine levels and myeloperoxidase activity in lung tissues were detected. Rat pulmonary microvascular endothelial cells (PMVECs) were stimulated by burn sepsis serum with or without 10 µM NAV-2729. The ARF6 activation, VE-cadherin expression, inflammasome activity, adapter protein apoptosis speck-like protein containing a caspase recruiting domain (ASC) specks and cytokines secretion were determined. Student's t test was used for comparison between two groups. Multiple comparisons among groups were performed by using analysis of variance, with Tukey's test for the post hoc test. RESULTS: NAV-2729 treatment attenuated burn sepsis induced lung injury and promoted survival of burn septic mice by preserving VE-cadherin expression in endothelial cell adherent junction and limited vascular hyperpermeability in lung tissues. Moreover, inflammatory cytokine expression and inflammatory injury in lung tissues were alleviated. Mechanistically, NAV-2729 enhanced vascular integrity by inhibiting ARF6 activation and restoring VE-cadherin expression in PMVECs. In addition, NAV-2729 inhibited ARF6-dependent phagocytosis of ASC specks, thus preventing inflammation propagation mediated by cell-to-cell transmission of ASC specks. CONCLUSIONS: ARF6 inhibition preserved vascular integrity by restoring expression of VE-cadherin and suppressed the spread of inflammation by affecting phagocytosis of ASC specks, thus protected against sepsis induced lung injury and improve survival of burn septic animals. The findings of this study implied potential therapeutics by which ARF6 inhibition can protect lung function from septic induced lung injury and improve outcomes in burn sepsis.

Pathophysiology of diabetic kidney disease and autophagy: A review.

Diabetic kidney disease (DKD) is one of the main complications of diabetic microangiopathy. The pathogenesis of DKD is very complex, including autophagy, inflammation, oxidative stress. Although a series of treatment intervention have achieved certain results in the treatment of diabetic nephropathy, still cannot reverse the kidney injury of diabetic nephropathy. The kidney is one of the most important organs of energy metabolism. Renal function is highly dependent on phagocytosis of mitochondria, and aberrant or defective autophagic mechanisms are central to the pathology of many renal diseases. Under high glucose conditions, mitochondrial fragments accumulate in the kidney, suggesting that mitochondrial clearance mechanisms may be attenuated with changes in mitochondrial transformation mechanisms. However, the exact mechanism of mitophagy regulation in DKD has not been elucidated. Recent advances in autophagy have renewed interest in these signaling pathways and molecules in the pathogenesis of DKD. Investigating autophagy and its associated signaling molecules may provide potential unique targets for therapeutic intervention in DKD.

Effect of Al Content on the Wear Evolution of Ti1-xAlxN-Coated Tools Milling Ti-6Al-4V Alloy.

Ti1-xAlxN coating is formed by replacing some Ti atoms in TiN with Al atoms, and their properties are closely related to Al content (0 < x < 1). Recently, Ti1-xAlxN-coated tools have been widely used in the machining of Ti-6Al-4V alloy. In this paper, the hard-to-machine material Ti-6Al-4V alloy is used as the study material. Ti1-xAlxN-coated tools are used for milling experiments. The evolution of the wear form and the wear mechanism of Ti1-xAlxN-coated tools are studied, and the influence of Al content (x = 0.52, 0.62) and cutting speed on tool wear are analyzed. The results show that the wear on the rake face changes from the initial adhesion and micro-chipping to coating delamination and chipping. Wear on the flank face varies from the initial adhesion and grooves to boundary wear, build-up layer, and ablation. The main wear mechanisms of Ti1-xAlxN-coated tools are dominated by adhesion, diffusion, and oxidation wear. Ti0.48Al0.52N coating protects the tool well and extends its service life.

Role of the TPX2/NCOA5 axis in regulating proliferation, migration, invasion and angiogenesis of breast cancer cells.

Breast cancer is a common malignant tumor in women. Increasing evidence has demonstrated that nuclear receptor coactivator 5 (NCOA5) and targeting protein for xenopus kinesin-like protein 2 (TPX2) serve vital roles in the progression of breast cancer. However, to the best of our knowledge, the molecular mechanisms underlying the involvement of TPX2/NCOA5 in the development of breast cancer are not fully understood at present. In the present study, the expression levels of NCOA5 and TPX2 were compared between paired non-tumor and tumor tissues of patients with breast cancer using the TNMplot tool. Expression differences of NCOA5 and TPX2 in human breast epithelial cell lines (MCF10A and MCF12A) and human breast cancer cell lines (MCF7 and T47D) were assessed via reverse transcription-quantitative PCR and western blotting. Additionally, proliferation, migration and invasion of breast cancer cells were determined via Cell Counting Kit-8, would healing and transwell assays. In vitro angiogenesis was determined using a tube formation assay. Furthermore, TPX2 was identified as a high-confidence NCOA5 interactor based on BioPlex network data sets. A co-immunoprecipitation assay was adopted to confirm the interaction between TPX2 and NCOA5. The present study revealed that TPX2 and NCOA5 were highly expressed in breast cancer cells. TPX2 interacted with NCOA5 and there was a positive association between TPX2 and NCOA5 expression. NOCA5 knockdown repressed the proliferation, migration, invasion and in vitro angiogenesis of breast cancer cells. In addition, TPX2 knockdown suppressed the proliferation, migration and invasion of breast cancer cells, and inhibited in vitro angiogenesis, and all of these effects were reversed following NCOA5 overexpression. In conclusion, NCOA5 was a downstream target of TPX2 in enhancing proliferation, migration, invasion and angiogenesis of breast cancer cells.

Assessment of disaster preparedness and related impact factors among emergency nurses in tertiary hospitals: descriptive cross-sectional study from Henan Province of China.

Background: The aim of this study was to investigate the current state of disaster preparedness and to determine associated factors among emergency nurses from tertiary hospitals in Henan Province of China. Methods: This multicenter descriptive cross-sectional study was conducted with emergency nurses from 48 tertiary hospitals in Henan Province of China between September 7, 2022-September 27, 2022. Data were collected through a self-designeds online questionnaire using the mainland China version of the Disaster Preparedness Evaluation Tool (DPET-MC). Descriptive analysis and multiple linear regression analysis were used to evaluate disaster preparedness and to determine factors affecting disaster preparedness, respectively. Results: A total of 265 emergency nurses in this study displayed a moderate level of disaster preparedness with a mean item score of 4.24 out 6.0 on the DPET-MC questionnaire. Among the five dimensions of the DPET-MC, the mean item score for pre-disaster awareness was highest (5.17 ± 0.77), while that for disaster management (3.68 ± 1.36) was the lowest. Female gender (B = -9.638, p = 0.046) and married status (B = -8.618, p = 0.038) were negatively correlated with the levels of disaster preparedness. Five factors positively correlated with the levels of disaster preparedness included having attended in the theoretical knowledge training of disaster nursing since work (B = 8.937, p = 0.043), having experienced the disaster response (B = 8.280, p = 0.036), having participated in the disaster rescue simulation exercise (B = 8.929, p = 0.039), having participated in the disaster relief training (B = 11.515, p = 0.025), as well as having participated in the training of disaster nursing specialist nurse (B = 16.101, p = 0.002). The explanatory power of these factors was 26.5%. Conclusion: Emergency nurses in Henan Province of China need more education in all areas of disaster preparedness, especially disaster management, which needs to be incorporated into nursing education, including formal and ongoing education. Besides, blended learning approach with simulation-based training and disaster nursing specialist nurse training should be considered as novel ways to improve disaster preparedness for emergency nurses in mainland China.

Analysis of bulk RNA-seq data from sepsis patients reveals sepsis-associated lncRNAs and targeted cell death-related genes contributing to immune microenvironment regulation.

Background: Sepsis is a life-threatening organ dysfunction syndrome that leads to the massive death of immune cells. Long non-coding RNAs (lncRNAs) have been reported to exert key regulatory roles in cells. However, it is unclear how lncRNAs regulate the survival of immune cells in the occurrence and development of sepsis. Methods: In this study, we used blood whole transcriptome sequencing data (RNA-seq) from normal controls (Hlty) and patients with uncomplicated infection (Inf1 P), sepsis (Seps P), and septic shock (Shock P), to investigate the fraction changes of immune cell types, expression pattern of cell death-related genes, as well as differentially expressed lncRNAs. Association network among these factors was constructed to screen out essential immune cell types, lncRNAs and their potential targets. Finally, the expression of lncRNAs and cell death genes in sepsis patients were validated by qRT-PCR. Results: In this study, we found fifteen immune cell types showed significant fraction difference between Hlty and three patient groups. The expression pattern of cell death-related genes was also dysregulated in Hlty compared with patient groups. Co-expression network analysis identified a key turquoise module that was associated with the fraction changes of immune cells. We then identified differentially expressed lncRNAs and their potential targets that were tightly associated with the immune cell dysregulation in sepsis. Seven lncRNAs, including LINC00861, LINC01278, RARA-AS1, RP11-156P1.3, RP11-264B17.3, RP11-284N8.3 and XLOC_011309, as well as their co-expressed cell death genes, were finally identified, and we validated two lncRNAs (LINC00861 and LINC01278) and four mRNA targets using qRT-PCR in sepsis samples. Conclusion: The global analysis of cell death-related genes in the occurrence and development of sepsis was carried out for the first time, and its expression regulation mode was displayed. The expression pattern of sepsis-associated lncRNAs were analyzed and identified, and the lncRNAs were significantly related to the change of immune cell proportion. We highlight the important roles of lncRNAs and their potential targets in the regulation of immune cell fraction changes during sepsis progression. The identified lncRNAs and their target genes may become new biomarkers and therapeutic targets of sepsis.

Regional citrate versus heparin anticoagulation for continuous renal replacement therapy in critically ill patients: A meta-analysis of randomized controlled trials.

INTRODUCTION: This study aimed to compare the efficacy and safety of citrate and heparin in continuous renal replacement therapy (CRRT) for critically ill patients. METHODS: Searched in PubMed, Embase, and Cochrane Library databases. RESULTS: Analyses showed that there no difference existed in mortality, metabolic alkalosis, circuit loss, and the number of transfused between the two groups (RR = 0.95, p = 0.40; RR = 1.73, p = 0.40; RR = 0.64, p = 0.09; RR = 1.05, p = 0.70). The filter life of the citrate group was longer than the heparin group (MD = 16.98, p < 0.0001). The risk of bleeding and heparin-induced thrombocytopenia was significantly lower in the citrate (RR = 0.32, p < 0.00001; RR = 0.55, p = 0.04). The citrate group was more susceptible to hypocalcemia (RR = 4.85, p = 0.0004). CONCLUSION: Citrate anticoagulant therapy should have priority for CRRT in most critically ill patients.

Preoperative CT-based peritumoral and tumoral radiomic features prediction for tumor spread through air spaces in clinical stage I lung adenocarcinoma.

OBJECTIVES: This study aims to develop and evaluate preoperative CT-based peritumoral and tumoral radiomic features to predict tumor spread through air space (STAS) status in clinical stage I lung adenocarcinoma (LUAD). MATERIALS AND METHODS: From June 2018 to December 2019, a retrospective diagnostic investigation was done. Patients with pathologically confirmed STAS status (N = 256) were eventually enrolled. The development cohort consisted of 191 patients (74.6%) chosen randomly in a 7:3 ratio, whereas the validation group consisted of 65 patients (25.4%). The performance of models was assessed using receiver operating characteristic analysis, accuracy, sensitivity, specificity, negative predictive values, and positive predictive values. RESULTS: The STAS positive status was found in 85 (33.2%) of the 256 patients (female: 53.2%; median [IQR] age: 62.0, [53.0-79.0] years), while the STAS negative status was found in 171 patients (66.8%) (female:50.6%; median [IQR] age: 62.0, [53.0-87.0] years). The combined TRS and PRS-15 mm model had an AUC of 0.854 (95% CI, 0.799-0.909) in the development cohort and 0.870 (95% CI, 0.781-0.958) in the validation cohort, indicating that the tumor radiomic signature (TRS) model and different peritumoral radiomic signature (PRS) models were used to build the optimal gross radiomic signature (GRS) model. The radiomic nomogram achieves superior discriminatory performance than GRS and clinical and radiological signatures (CRS), with an AUC of 0.871 (95% CI, 0.820-0.922) in the development cohort and AUC of 0.869 (95% CI, 0.776-0.961) in the validation cohort. Based on the Akaike information criterion (AIC) and decision curve analysis (DCA), the radiomic nomogram provided greater clinical predictive capacity than clinical or any radiomic signatures alone. CONCLUSION: In conclusion, we discovered that peritumoral characteristics were substantially related to STAS status. This study revealed the unit of radiomic signature and clinical signatures may have a better performance in STAS status.

The effect of a coix seed oil injection on cancer pain relief.

BACKGROUND: Pain is one of the most commonly reported symptoms in patients with advanced cancer, but is still less than optimally treated. The effect of traditional Chinese medicine in cancer pain treatment is nowadays getting more and more attention. OBJECTIVE: To investigate the effect of a coix seed oil injection on cancer pain relief in a cancer center in a tertiary hospital in China. METHODS: Patients in the treatment group received a coix seed oil injection for 2 weeks, while patients in the control group received equivalent 0.9% saline. The numeric rating scale was used to assess the pain level. The Quality of Life Questionnaire-Core 30 was used to assess life quality. The adverse drug reactions during the treatment process were observed. RESULTS: Patients in the coix seed treatment group had significantly superior efficacy on pain control over those in the control group. Coix seed therapy significantly improved patients' scores reflecting by the Quality of Life Questionnaire-Core 30 (QLQ-C30) scale. In addition, the occurrence of adverse reactions such as constipation and nausea in the treatment group was significantly lower than that in the control group. CONCLUSION: The coix seed oil injection effectively reduced the pain level of cancer patients, significantly improved their life quality, and had no obvious adverse effects.

MicroRNA-873 acts as a tumor suppressor in esophageal cancer by inhibiting differentiated embryonic chondrocyte expressed gene 2.

Esophageal cancer is one of the most common digestive malignant diseases worldwide and emerging evidences revealed that microRNAs (miRNAs) were implicated in the development and progression of esophageal cancer. However, the expression level and biological function of microRNA-873(miR-873) in esophageal cancer are still largely elusive. In this study, we investigated the expression and biological roles of miR-873 in human esophageal cancer. Our results revealed that miR-873 was significantly underexpressed in esophageal cancer tissues and cell lines when compared with the para-tumor tissue and primary human esophageal epithelial cells. Furthermore, overexpression of miR-873 could remarkably inhibit esophageal cancer cell growth, migration and invasion. Moreover, we validated differentiated embryonic chondrocyte expressed gene 2 (DEC2) as a direct target of miR-873 which could reverse the repressive effects of miR-873 on esophageal cancer cell. In summary, our investigation demonstrated that miR-873 was underexpressed in esophageal cancer and might act as a tumor suppressor gene by directly targeting DEC2.

Leptin/OB-R pathway promotes IL-4 secretion from B lymphocytes and induces salivary gland epithelial cell apoptosis in Sjögren's syndrome.

Sjögren's syndrome (SjS) is a chronic autoimmune epithelitis in which cell apoptosis promotes the formation of inflammatory lesions. We used immunohistochemistry and TUNEL to assay B cell infiltration and apoptosis in salivary gland tissue from 16-week-old NOD/LtJ mice with SjS. In co-cultures of primary salivary glandepithelial cells (SGECs) and spleen B cells, we assessed SGEC viability and apoptosis using CCK8 assays and flow cytometry. ELISAs were employed to assess cytokine levels in culture medium. Leptin protein, leptin receptor (OB-R), pro- and anti-apoptotic proteins, and Jak2/Stat3/ERK signaling molecules were analyzed using western blotting. B cell infiltration and salivary gland apoptosis were increased in salivary tissue from mice with SjS. Leptin treatment had no effect on cell viability or apoptosis among B cells and primary SGECs. B cell and SGEC co-culture systems showed that leptin increased apoptosis induced by B lymphocytes, reduced SGEC cell viability, and promoted IL-4 secretion from B cells. This suggests Leptin/OB-R signaling stimulates B cells-induced SGEC apoptosis via IL-4 secretion and OB-R-Jak2-Stat3 activation.

Leptin/OB-R signaling is elevated in mice with Sjögren's syndrome and is implicated in disease pathogenesis.

Sjögren's syndrome (SjS) is a systemic autoimmune disease resulting in a severe dry mouth and dry eyes. Currently, care for patients with SjS is palliative, as no established therapeutics target the disease directly, and its pathogenetic mechanisms are uncertain. Leptin activates B cells to induce the secretion of proinflammatory and anti-inflammatory cytokines and is elevated in several autoimmune diseases. In this study, we found the expression of leptin and its receptor OB-R in mouse models of SjS are elevated both locally and systemically during SjS progression. Recombinant serotype 2 adeno-associated viral (rAAV2) vectors expressing either OB-R shRNA (rAAV2-shOB-R) or none (rAAV2-null) were injected into 4 or 16 week-old BALB/c NOD/LtJ (NOD) mice and resulted in a modest reduction in glandular inflammation in the SjS model. In conclusion, Leptin/OB-R signaling may be pathogenically involved in SjS and may serve as a new marker and a potential therapeutic target.

Effect of Glucose Concentration on Electrochemical Corrosion Behavior of Pure Titanium TA2 in Hanks' Simulated Body Fluid.

Titanium and its alloys have been widely used as implant materials due to their excellent mechanical property and biocompatibility. In the present study, the effect of glucose concentration on corrosion behavior of pure titanium TA2 in Hanks' simulated body fluid is investigated by the electrochemical impedance spectrum (EIS) and potentiodynamic polarization methods. The range of glucose concentrations investigated in this research includes 5 mmol/L (limosis for healthy people), 7 mmol/L (after diet for healthy people), 10 mmol/L (limosis for hyperglycemia patient), and 12 mmol/L (after diet for hyperglycemia patient), as well as, 15 mmol/L and 20 mmol/L, which represent different body fluid environments. The results indicate that the pure titanium TA2 demonstrates the best corrosion resistance when the glucose concentration is less than 10 mmol/L, which shows that the pure titanium TA2 as implant material can play an effective role in the body fluids with normal and slight high glucose concentrations. Comparatively, the corrosion for the pure titanium implant is more probable when the glucose concentration is over 10 mmol/L due to the premature penetration through passive film on the material surface. Corrosion defects of pitting and crevice exist on the corroded surface, and the depth of corrosion is limited to three microns with a low corrosion rate. The oxidation film on the surface of pure titanium TA2 has a protective effect on the corrosion behavior of the implant inner material. The corrosion behavior of pure titanium TA2 will happen easily once the passive film has been penetrated through. The corrosion rate for TA2 implant will accelerate quickly and a pure titanium implant cannot be used.

Targeting druggable enzymome by exploiting natural medicines: An in silico-in vitro integrated approach to combating multidrug resistance in bacterial infection.

CONTEXT: Antibiotic resistance is a major clinical and public health problem. Development of new therapeutic approaches to prevent bacterial multidrug resistance during antimicrobial chemotherapy has thus been becoming a primary consideration in the medicinal chemistry community. OBJECTIVE: We described a new strategy that combats multidrug resistance by using natural medicines to target the druggable enzymome (i.e., enzymatic proteome) of Staphylococcus aureus. MATERIALS AND METHODS: A pipeline of integrating in silico analysis and in vitro assay was purposed to identify antibacterial agents from a large library of natural products with diverse structures, high drug-likeness, and relatively low flexibility, with which a systematic interactome of 826 natural product candidates with 125 functionally essential S. aureus enzymes was constructed via a high-throughput cross-docking approach. The obtained docking score matrix was then converted into an array of synthetic scores; each corresponds to a natural product candidate. By systematically examining the docking results, a number of highly promising candidates with potent antibacterial activity were suggested. RESULTS: Three natural products, i.e., radicicol, jorumycin, and amygdalin, have been determined to possess strong broad-spectrum potency combating both the drug-resistant and drug-sensitive strains (MIC value <10 µg/ml). In addition, some natural products such as tetrandrine, bilobalide, and arbutin exhibited selective inhibition on different strains. DISCUSSION AND CONCLUSION: Combined quantum mechanics/molecular mechanics analysis revealed diverse non-bonded interactions across the complex interfaces of newly identified antibacterial agents with their putative targets GyrB ATPase and tyrosyl-tRNA synthetase.

Overexpression of CRY1 protects against the development of atherosclerosis via the TLR/NF-κB pathway.

It has been demonstrated that the circadian clock system could be a potential factor involved in inflammation and the progression of atherosclerosis. A previous study has reported that cryptochrome 1 (CRY1), which is a core clock component, is associated with regulating proinflammation. However, whether CRY1 is involved in atherosclerosis is currently unknown. In the present study, we aimed to explore the role of CRY1 in regulating atherosclerosis in apolipoprotein E (ApoE)-deficient mice and the underlying molecular mechanism. We found that CRY1 mRNA expression was significantly decreased in atherosclerotic patients compared to the healthy subjects. Overexpression of CRY1 in the mouse model of atherosclerosis by adenovirus-mediated gene transfer significantly decreased the expression of proinflammatory factors including tumor necrosis factor-α (TNF-α), interleukin (IL)-1, IL-6, and macrophage inflammatory protein-1α (MIP-1α). In addition, the adhesion molecules including vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin, were also downregulated by CRY1 overexpression. Furthermore, the plaque area of the aortic sinus and the concentrations of total cholesterol (TC), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C) were also decreased in the atherosclerotic mice by CRY1 overexpression. Moreover, overexpression of CRY1 significantly decreased the protein levels of toll-like receptor (TLR) 2, TLR4 and phosphorylated p65 (p-p65). Additionally, the results of luciferase reporter assay exhibited that CRY1 overexpression was capable of inhibiting the activation of nuclear factor-kappa B (NF-κB). Taken together, our results suggest that overexpression of CYR1 relieves the development of atherosclerosis that may be associated with regulating the TLR/NF-κB pathway.

siRNA-mediated silencing of Wnt5a regulates inflammatory responses in atherosclerosis through the MAPK/NF-κB pathways.

Previous studies have demonstrated that the aberrant expression of Wnt5a occurs in atherosclerotic lesions. However, the precise role of Wnt5a in the pathogenesis of atherosclerosis remains largely unknown. The present study was undertaken to determine whether the RNA interference of Wnt5a in vivo by adenovirus (Ad)-mediated small interfering RNA (siRNA) transfection is capable of inhibiting the progression of atherosclerosis. Recombinant adenovirus carrying siRNA targeting Wnt5a (Ad-Wnt5a siRNA) was designed. Male apolipoprotein E-deficient (ApoE(-/-)) mice were fed a high-fat diet to induce the pathogenesis of atherosclerosis. Mice were randomly divided into 3 groups (n=15 in each group): the mock group, which received treatment with phosphate-buffered saline (PBS); the Ad-NC group, which received treatment with Ad-non-specific siRNA; and the Ad-Wnt5a siRNA group, which received treatment with Ad-Wnt5a siRNA. Treatment with Ad-Wnt5a siRNA markedly inhibited the mRNA and protein expression of Wnt5a in the aortic tissues. The knockdown of Wnt5a had no significant effect on blood lipid levels, but it suppressed atherosclerotic development and increased plaque stability, which was determined by hematoxylin and eosin staining, picrosirius red staining and Oil Red O staining. Furthermore, the mRNA and protein expression of inflammatory cytokines, including monocyte chemotactic protein-1 (MCP-1), cyclooxygenase-2 (COX-2), matrix metalloproteinase (MMP)-2 and MMP-9 was significantly downregulated in the Ad-Wnt5a siRNA group. In addition, the knockdown of Wnt5a inhibited the nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. These results demonstrate that Ad-mediated Wnt5a silencing in vivo attenuates the development of atherosclerotic disease by reducing inflammatory mediators involved in the MAPK/NF-κB pathways.

Role of autophagy and apoptosis in wound tissue of deep second-degree burn in rats.

OBJECTIVES: The pathogenesis of burn wound progression is poorly understood. Contributing factors include continuous loss of blood perfusion, excessive inflammation, and elevated apoptosis levels in wound tissue. Macroautophagy (here referred to simply as "autophagy") is associated with many chronic diseases. The authors hypothesized that autophagy is involved in burn wound progression in a rat model of deep second-degree burn. METHODS: Deep second-degree burns were modeled using a brass rod heated to 100°C applied for 6 seconds to the back skin of Wistar rats. Full-thickness biopsies were obtained from burned and nonburned controls at several times postburn. Western blotting and immunohistochemical (IHC) staining determined expression of the autophagy markers Light Chain 3 (LC3) and beclin-1. Apoptosis was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and laser Doppler flowmetry (LDF)-measured tissue perfusion. Myeloperoxidase (MPO) activity assay measured inflammation. Hematoxylin and eosin (H&E) and Masson's trichrome staining-determined pathology and wound depth. RESULTS: The LC3 and beclin-1 protein level in burn wounds decreased to one-fourth of normal levels (p<0.01) over 24 hours and then began to increase but still did not reach their normal level. TUNEL-positive cells in burn wounds were 3.7-fold (p<0.01) elevated over 48 hours and then decreased slightly, yet still remained higher than in normal skin. The burn wound progressed in depth over 72 hours. In addition, significant decrease in LDF values and upregulation of MPO activity were observed. Enhanced LC3-positive cells were observed in the deep dermal layer of burn wounds as shown by IHC staining. CONCLUSIONS: A reduction in autophagy and blood flow and an increase in apoptosis and inflammation were observed in burn wounds early during the course of burn injury progression. This suggests that autophagy, complemented by apoptosis, play important roles in burn progression. Enhanced autophagy in the deep dermis may be a prosurvival mechanism against ischemia and inflammation after burn injury.