Soluble elastin fragments in serum are elevated in aortic dissection

OBJECTIVES: We aimed to establish an enzyme-linked immunosorbent assay (ELISA) for measuring soluble elastin fragments (sELAF) in serum and to reveal its usefulness in diagnosing acute aortic dissection. BACKGROUND: Acute aortic dissection is a life-threatening disease of the aorta. However, the diagnosis is still frequently missed, especially at onset. The establishment and clinical availability of simplified laboratory test(s) to help diagnose and screen acute aortic dissection patients is therefore urgently needed. METHODS and RESULTS: An ELISA to measure sELAF in serum was developed using the newly created double monoclonal antibodies which recognize the different epitopes of human aortic elastin. Twenty-five acute aortic dissection patients, 50 patients with acute myocardial infarction, and 474 healthy individuals were enrolled in the study. The sELAF levels from healthy subjects gradually increased with aging. When the cutoff point for positivity was set at the mean + 3SD above the mean of those in healthy subjects at each age, 16 acute aortic dissection patients (64.0%) were found to be positive, while only one acute myocardial infarction patient was positive (2.0%). Acute aortic dissection patients with either an open or a partially open pseudolumen were found to be 88.9% positive for sELAF, while those with its early closure was 0% positive. The difference in the sELAF levels between acute aortic dissection patients with and without a thrombotic closure of false lumen was significant (60.3 +/- 15.6 vs 135.4 +/- 53.2 ng/ml, p < 0.005). CONCLUSIONS: The sELAF level in serum may be a useful marker for helping both diagnose and screen acute aortic dissection, while also helping distinguish acute aortic dissection from acute myocardial infarction.     

Novel reporter system to monitor early stages of the hepatitis B virus life cycle

A recombinant hepatitis B virus (HBV) expressing NanoLuc (NL) (HBV/NL) was produced by cotransfecting a plasmid containing a 1.2‐fold HBV genome carrying the NL gene with a plasmid bearing a packaging‐defective 1.2‐fold HBV genome into a human hepatoma cell line, HepG2. We found that NL activity in HBV/NL‐infected primary hepatocytes or sodium taurocholate cotransporting polypeptide‐transduced human hepatocyte‐derived cell lines increased linearly for several days after infection and was concordant with HBV RNA levels in the cells. Treatment of the virus‐infected cells with HBV inhibitors reduced NL activity in a dose‐dependent manner. Detection of HBV/NL infection, monitored by NL activity, was highly sensitive and less expensive than detection using the conventional method to evaluate HBV infection. In addition, because we also studied host factors, this system is applicable not only for studying the HBV life cycle, but also for exploring agent(s) that regulate HBV proliferation.     

Intravesical administration of exogenous microRNA-145 as a therapy for mouse orthotopic human bladder cancer xenograft

We previously reported that the level of microRNA (miR)-145 is attenuated in human bladder cancer cells. In this current study, we investigated whether intravesical administration of miR-145 could be a potential therapeutic strategy for controlling bladder cancer by using an orthotopic human bladder cancer xenograft model. Following transfection of 253J B-V cells with miR-145, the effects of the ectopic expression of miR-145 were examined by performing MTT, Western blotting analysis, Hoechst33342 staining, and wound healing assay in vitro. Also, a mouse orthotopic human bladder cancer model was established by inoculating 253J B-V cells into the bladder wall of mice. The anti-cancer effects of intravesical injections of miR-145 into these mice were then assessed. Transfection of 253J B-V cells with miR-145 induced apoptosis and suppression of cell migration in vitro. Western blotting showed that the levels of c-Myc, socs7, FSCN1, E-cadherin, β-catenin, and catenin δ-1 were decreased and that the PI3K/Akt and Erk1/2 signaling pathways were increased in compensatory fashion. In vivo, mice treated with miR-145 showed 76% inhibition of tumor growth, with a significant prolongation of animal survival (p = 0.0183 vs. control). Western blotting showed that both apoptosis and cell motility-related genes were significantly decreased as seen in vitro. Furthermore, PI3k/Akt and Erk1/2 signaling pathways, which were activated in a compensatory manner in vitro, were decreased in vivo. Intravesical administration of exogenous miR-145 was thus concluded to be a valid therapy for bladder cancer in this human bladder cancer xenograft model.