Label-free quantitative proteomic analysis of serum exosomes in mice with thoracic aortic aneurysm.

OBJECTIVE: Thoracic aortic aneurysm (TAA) is a cardiovascular disease with high morbidity and mortality. However, the causes and mechanisms of TAA are not fully understood. Serum exosomes from mice with TAA were used to explore the markers associated with this disease. METHODS: C57BL/6 mice were divided into three groups and given ordinary drinking water, ordinary drinking water plus a saline osmotic pump, or drinking water containing ß-aminopropionitrile (BAPN) (1 g/kg/d) plus an angiotensin II (Ang II) (1 µg/kg/min) osmotic pump. Haematoxylin and eosin staining of thoracic aortic tissues was performed. The basic characteristics of exosomes were analysed. Differentially expressed proteins (DEPs) were identified by LC‒MS/MS. Protein‒protein networks and enrichment analysis were used to explore possible molecular mechanisms. RESULTS: The present study elucidated the protein expression profile of serum exosomes in mice with TAA induced by BAPN combined with Ang II. In this work, the expression of a total of 196 proteins was significantly dysregulated in serum exosomes of mice with TAA, with 122 proteins significantly upregulated and 74 proteins markedly downregulated. Notably, Haptoglobin (Hp) and Serum amyloid p-component (Sap) identified based on the PPI network were significantly upregulated and have been strongly linked to cardiovascular disease. Interestingly, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the upregulated and downregulated proteins were involved in the complement and coagulation cascade pathways. CONCLUSIONS: This study showed that the identified DEPs have potential as biomarkers for the diagnosis of TAA and provided a more comprehensive understanding of the pathophysiological mechanisms of TAA.

Hybrid Repair of Complicated Type B Aortic Dissection Aneurysm With Dissected Kommerell Diverticulum Without Thoracotomy.

PURPOSE: The coincidence of aberrant right subclavian artery (ARSA) and Kommerell diverticulum (KD) with type B aortic dissection (TBAD) is a rare but dangerous disease. Currently, there are no well-established guidelines for treatment. Most authors seem to agree that surgical treatment is warranted. However, a hybrid repair technique as we performed is flexible, and a promising approach should be considered. CASE REPORT: Here, we summarized a case report of successful single-stage hybrid repair of a complicated TBAD combined with ARSA and KD without thoracotomy. CONCLUSION: Hybrid repair is a flexible and promising technique that has the potential to replace most open operation procedures in the future with a developed technique and more evidence-based medicine. CLINICAL IMPACT: As for ARSA and KD with TBAD patients, open surgical repair has been historically the treatment of choice; however, hybrid repair without thoracotomy means less invasion, simpler operation and faster recovery, which provides a flexible and promising technique that has the potential to replace most open operation procedures in the future with more evidence-based medicine.

The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC.

The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome. This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect. A lentiviral vector was used to stably transfect human umbilical vein endothelial cells (HUVECs) with PGI2S alone (HUVEC-PGI2S) or both PGI2S and tPA (HUVEC-PGI2S-tPA). Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells. The enzyme-linked immuno sorbent assay (ELISAs) demonstrated that the anticoagulation components, ATIII and PLG, were up-regulated and coagulation factor FVIII was down-regulated in both cell lines. QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA, PKC, and PTGIR were up-regulated in both cell lines, but MAPK expression was not altered in either cell line. However, cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced G1 phase arrest. HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis.

In vitro activity of ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam alone or in combination with polymyxin B against carbapenem resistant Acinetobacter baumannii.

Nosocomial infection caused by Carbapenem-Resistant Acinetobacter baumannii (CR-A. baumannii) has become a challenge in clinical practice. Acting as the last resort antibacterial agents for the treatment of CR-A. baumannii infection, polymyxins have high risk of nephrotoxicity and poor clinical efficacy. Ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam are three ß-lactam/ß-lactamase inhibitor combination complexes that newly approved by the Food and Drug Administration for the treatment of carbapenem-resistant Gram-negative bacterial infection. In this study, we analyzed the in vitro activity of those novel antibacterial agents alone or in combination with polymyxin B against the CR-A. baumannii obtained from a Chinese tertiary hospital. Our results suggest that those novel antibacterial agents should not be used alone for the treatment of CR-A. baumannii infection, as they cannot prevent the regrowth of bacteria at the clinical achievable blood concentration. Imipenem/relebactam and meropenem/vaborbactam should not be used as the substitutes of imipenem and meropenem for polymyxin B based combination therapy against CR-A. baumannii, since they have no edge over imipenem and meropenem on antibacterial activity when in combination with polymyxin B. Ceftazidime/avibactam may be more suitable than ceftazidime for polymyxin B based combination therapy against CR-A. baumannii, as it has a higher synergistic rate with polymyxin B, and the antibacterial activity of ceftazidime/avibactam is much higher than that of ceftazidime when tested in combination with polymyxin B. Ceftazidime/avibactam may also be the better choice than imipenem and meropenem for polymyxin B based combination therapy against CR-A. baumannii, as it has a higher synergistic rate with polymyxin B.

In vitro analysis of synergistic combination of polymyxin B with 12 other antibiotics against MDR Acinetobacter baumannii isolated from a Chinese tertiary hospital.

In clinical practice, polymyxins are suggested to be used in combination with other antibiotics for improving their antibacterial efficacy and preventing the emergency of antibiotic-resistant strains. However, even though synergistic combination of polymyxin B with many antibiotics have been confirmed in various studies with different bacterial species and analyzing methods, which antibiotic is the best option for combination therapy of polymyxin B against MDR A. baumannii remains uncertain. In this study, we systematically analyzed the synergistic combination of polymyxin B with 12 other antibiotics against MDR A. baumannii isolated from a Chinese tertiary hospital using the checkerboard assay. The results suggest that, for polymyxin B-based combination therapy against MDR A. baumannii as characterized in this hospital, cefperazone-sulbactam may be the best partner, since it has the highest synergistic rate and the best synergistic effect with polymyxin B. Minocycline, imipenem, meropenem, ceftazidime, cefepime, amikacin and sulfamethoxazole also have some synergistic effects with polymyxin B, but piperacillin-tazobactam, ciprofloxacin, levofloxacin and tobramycin show no synergism. None of these 12 antibiotics has an antagonistic effect when combined with polymyxin B.

Synergistic antibacterial and anti-biofilm activities of resveratrol and polymyxin B against multidrug-resistant Pseudomonas aeruginosa.

Bacterial infection caused by multidrug-resistant Pseudomonas aeruginosa has become a challenge in clinical practice. Polymyxins are used as the last resort agent for otherwise untreatable Gram-negative bacteria, including multidrug-resistant P.aeruginosa. However, pharmacodynamic (PD) and pharmacokinetic (PK) data on polymyxins suggest that polymyxin monotherapy is unlikely to generate reliably efficacious plasma concentrations. Also, polymyxin resistance has been frequently reported, especially among multidrug-resistant P.aeruginosa, which further limits its clinical use. A strategy for improving the antibacterial activity of polymyxins and preventing the development of polymyxin resistance is to use polymyxins in combination with other agents. In this study, we have demonstrated that resveratrol, a well tolerated compound, has synergistic effects when tested in vitro with polymyxin B on antibacterial and anti-biofilm activities. However, its' systemic use is limited as the required high plasma levels of resveratrol are not achievable. This suggests that it could be a partner for the combination therapy of polymyxin B in the treatment of topical bacterial infection caused by MDR P.aeruginosa.

Combined prognostic value of the cancer stem cell markers CD47 and CD133 in esophageal squamous cell carcinoma.

BACKGROUND: Treatments based on the inhibition of pivotal signals of cancer stem cells (CSCs) are on a promising track. Recent studies have shown that targeting CSCs with broader immune-based therapeutic methods, for example, the anti-CD47 treatment, may serve as a more potent strategy for eliminating these intractable cells. We aimed to explore the prognostic effects of CD47/CD133 and the potential therapeutic significance of CD47 in esophageal squamous cell carcinoma (ESCC). METHODS: Immunohistochemistry was employed to identify the characteristics of CD47 and CD133 in 26 pairs of tumor tissues and adjacent non-tumor tissues and 136 ESCC tissues. Kaplan-Meier analysis and Cox proportional hazards models were built for estimating the prognostic values of CD47 and CD133 expression and their combined stemness index. Sphere formation assays were undertaken to explore the effects of CD47 inhibition on primary human ESCC CSCs. RESULTS: Results conclude that CD47 and CD133 expression is increased in tumor tissues as compared to adjacent non-tumor tissues. A positive correlation between CD47/CD133 expression and differentiation was found in 136 ESCC patients. Survival analysis indicated that patients with high CD47 or CD133 expression exhibited poor overall survival and progression-free survival (PFS). The combination of high CD47 and CD133 expression was a reliable independent prognostic factor for both OS (HR = 1.940, 95% CI = 1.399-2.690, P < 0.0001) and progression-free survival (HR = 1.883, 95% CI = 1.384-2.562, P < 0.0001). Notably, CD47+ CD133+ ESCC cells were observed to possess the characteristics of CSCs, and anti-CD47 treatment veritably eliminated the CSCs pool. CONCLUSIONS: The stemness index determined by the expression of CD47 and CD133 is a promising prognostic predictor, and CD47 is a potential therapeutic target for CSCs in ESCC patients.

GOLPH3 promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma via mTOR and Wnt/ß­catenin signal activation.

The authors' previous study demonstrated that Golgi phosphoprotein 3 (GOLPH3) was significantly overexpressed in esophageal squamous cell carcinoma (ESCC), correlating with poor patient survival. In the present study, GOLPH3 stable overexpression and knockdown KYSE­140 cell lines were constructed. Cell proliferation, colony formation, cell cycle progression and tumorigenesis assays were performed. The results revealed that GOLPH3 promoted ESCC cell growth and proliferation. The effects of GOLPH3 on the mechanistic target of rapamycin (mTOR) and Wnt/ß­catenin signaling pathways were investigated using western blot analyis and dual­luciferase reporter assays, and were observed to be activated in cells with GOLPH3 overexpression. Furthermore, overexpression of GOLPH3 resulted in the downregulation of p21 protein, upregulation of cyclin D1 and increased retinoblastoma­associated protein phosphorylation, consequently leading to accelerated cell cycle progression. In addition, GOLPH3 knockdown resulted in reversed effects. The results of the current study suggest that GOLPH3 serves an important role in promoting tumorigenicity of ESCC via mTOR and Wnt/ß­catenin signaling pathway activation.

MicroRNA Expression Profile and the Role of miR-204 in Corneal Wound Healing.

PURPOSE: MicroRNAs (miRNAs) are endogenous short chain (∼ 22-nucleotide) noncoding RNAs that inhibit protein translation through binding to target mRNAs. Recent studies have implied that miRNAs play a regulatory role in corneal development. Here we profile their involvement in corneal epithelial renewal, develop an miRNA-target network that affects wound healing outcome, and investigate the function of miR-204 in this response. METHODS: NanoString nCounter technology and bioinformatics analyzed miRNA expression levels and their targets during mouse corneal epithelial wound healing. Real-time RT-PCR was performed to detect miR-204 expression in mouse corneal epithelium. Human corneal epithelial cells (HCECs) were transfected with miR-204 using transfection reagent. MTS (3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, inner salt) and a scratch wound-healing assay evaluated the effects of miR-204 expression on HCEC proliferation and migration, respectively. Cell cycle analysis was performed by flow cytometry. Expression of sirtuin 1 (SIRT1) was determined by Western blot analysis. RESULTS: Fifteen miRNAs were dramatically downregulated, whereas 14 other miRNAs were markedly upregulated during corneal wound healing. Expression of miR-204 fell the most during this process. Transfection of miR-204 into HCECs led to a significant decline in cell proliferation and induced cell cycle G1-arrest. Furthermore, in these cells, miR-204 also inhibited migration. Sirtuin 1 was confirmed as a target of miR-204. CONCLUSIONS: During mouse corneal epithelial wound healing, a complex miRNA-gene network was resolved that is modulated by changes in miR-204 expression. Downregulation of this miRNA appears to be an essential response to injury since its decline promotes human corneal epithelial cell proliferation and migration. Therefore, miR-204 could be a biomarker of this process.

RASSF8 downregulation promotes lymphangiogenesis and metastasis in esophageal squamous cell carcinoma.

Lymphatic vessels are the major routes of human esophageal squamous cell carcinoma (ESCC) metastasis. Tumor cells secrete pro-lymphangiogenic factors to induce new lymphatic vessels, promoting lymph node metastasis. In this study, we show that RAS association domain family 8 (RASSF8) expression in ESCC clinical samples was inversely correlated with lymph node metastasis and patients survival. Tumor cells with low RASSF8 expression had higher apparent migratory ability, and promoted and lymphangiogenesis both in vitro and in vivo. RASSF8 downregulation enhanced VEGF-C expression and caused subcellular redistribution of p65 in ESCC. Our results show that RASSF8 acts as a tumor suppressor in ESCC and is a potential therapeutic target for preventing lymph node metastasis.