[Role of brain and muscle ARNT-like protein 1 in the rat periodontitis-induced liver injury model].

Objective: Brain and muscle ARNT-like protein 1 (BMAL1) is a core component of hepatocyte molecular clock and plays an important role in the regulation of other related rhythmic genes in the body through a transcriptional-translational feedback loop in molecular circadian oscillations. Therefore, the aim of this study was to investigate the role of BMAL1 in the rat periodontitis-induced liver injury. Methods: Twelve male Wistar rats were divided into the control group and the periodontitis group according to the random number table method. The rats in the control group were untreated. The periodontitis models were established by ligating the necks of the bilateral maxillary first molars in the periodontitis group rats. After 8 weeks, periodontal clinical indexes of rats in both groups were examined and executed. Micro-CT scans of the maxilla were performed and levels of the alveolar bone resorption were analyzed. Pathological changes in periodontal and liver tissue of rats in two groups were detected by HE and oil red O staining. Biochemical kits were used to detect glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), total cholesterol (TC) and triglycerides (TG) in serum. The gene and protein expression levels of BMAL1, nuclear factor kappa-B (NF-κB) and tumor necrosis factor-α (TNF-α) in liver tissue were measured by real time fluorescent quantitative-PCR (qRT-PCR), immunohistochemistry (IHC) and Western blotting (WB) assays. Apoptosis was detected in liver tissues by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) kit staining. Results: The results of HE staining of maxillary first molars and micro-CT results of maxillary bones showed that alveolar bone resorption was significant in the periodontitis group of rats. The liver histopathology results showed infiltrated inflammatory cells in the liver tissue, disorganized liver cords and a large number of lipid droplets formed in the hepatocytes of the periodontitis group compared with the control group. The results of serum biochemical assay showed that the levels of GOT [(62.77±2.59) U/L], GPT [(47.54±1.04) U/L], TC [(3.19±0.23) mmol/L] and TG [(1.11±0.09) mmol/L] in the serum of rats with periodontitis were significantly higher than that in the control group respectively [GOT: (38.66±2.47) U/L, GPT: (31.48±1.57) U/L, TC: (1.60±0.05) mmol/L and TG: (0.61±0.09) mmol/L](P=0.003, P=0.001, P=0.002, P=0.038). qRT-PCR results showed that the mRNA expression level of BMAL1 was significantly decreased in liver tissue of the periodontitis group [(0.60±0.04)%] compared to the control group [(1.01±0.07)%] (t=4.80, P=0.009), while the mRNA expression levels of NF-κB and TNF-α [(1.62±0.12)%, (2.69±0.16)%] were significantly increased compared to the control group [(1.00±0.03)%, (1.03±0.16)%] (P=0.008, P=0.002); IHC results showed that the protein expression level of BMAL1 in liver tissue of the periodontitis group (averaged optical density, AOD) (11.58±2.15) was down-regulated compared to the control group (AOD) (22.66±1.67) (P=0.015), while NF-κB and TNF-α (AOD) (31.77±2.69, 24.31±2.32) were up-regulated compared to the control group (AOD) (19.40±1.82, 11.92±0.94) (P=0.019, P=0.008). WB results showed that the protein expression level of BMAL1 in liver tissue was down-regulated in the periodontitis group [(0.63±0.10)%] compared to the control group [(1.00±0.06)%] (t=3.19, P=0.033), while NF-κB and TNF-α [(1.61±0.12)%, (2.82±0.23)%] were up-regulated compared to the control group [(1.00±0.12)%, (1.00±0.11)%] (P=0.022, P=0.002). TUNEL staining showed increased apoptotic cells in the liver tissue of the periodontitis group of rats compared to the control group. Conclusions: Periodontitis may induce liver injury by down-regulating the BMAL1 expression levels in liver tissue, which in turn activates NF-κB signaling molecules, leading to the elevated levels of inflammation and apoptosis in rat liver.

Expression of SIRT1 gene in human lung cancer lines enhances their sensitivity to the anticancer effects of cisplatin.

OBJECTIVE: Lung cancer is one of the lethal cancers and one of the major causes of cancer-related mortalities across the globe. SIRT1 gene has been reported to be involved in the progression and tumorigenesis of several types of cancers. However, the role of SIRT1 gene is in the progression of lung cancer is poorly understood. Against this backdrop, the present study was designed to investigate the expression of SIRT1 gene in different lung cancer cell lines. Moreover, the relation between the expression of this gene and the sensitivity of lung cancer cell lines to the anticancer effects of cisplatin was also investigated. MATERIALS AND METHODS: Expression of SIRT1 gene was determined by quantitative RT-PCR. Protein expression was examined by Western blotting. Anti-proliferative activity was determined by MTT and colony formation assay. Apoptotic populations were determined by annexin V/IP staining and flow cytometry. RESULTS: The results revealed that NCI-H125 showed lowest, NCI-H226 showed moderate, while as NCI-H358 exhibited the highest expression of SIRT1. The three differentially SIRT1 expressing cancer cell lines were subjected to cisplatin treatment. It was observed that cisplatin exhibited the highest anticancer activity against NCI-H125 (IC50, 1.25 µM) and lowest against NCI-H358 (IC50, 4.5 µM). Moreover, cisplatin leads to highest inhibition of colony formation and apoptosis in NCI-H125 and lowest against NCI-H358. CONCLUSIONS: Expression of SIRT1 gene determines the sensitivity of lung cancer cells to anticancer effects of cisplatin. This work will pave for understanding the role of SIRT1 gene in cancer progression.

MiR-370 promotes cell death of liver cancer cells by Akt/FoxO3a signalling pathway.

OBJECTIVE: MicroRNAs have emerged as key regulators in cancer cell biology. In the present study, we investigate the role and the involving mechanism of miR-370 in the progression of liver cancers. MATERIALS AND METHODS: MiR-370 levels were detected by real-time PCR assay. Cell proliferation of HepG2, MHCC-97H and SMMC-7721 was determined by MTT assay. PI staining was detected by FACS analysis. Colony formation was used to test liver cancer cell growth. FoxO3a and Akt expression was determined by western blotting analysis. RESULTS: MiR-370 level was significantly down-regulated in liver cancer cells. Functional analysis revealed that miR-370 mimics suppressed cell proliferation of liver cancer cells, while transfection with miR-370 inhibitor increased cell proliferation of liver cancer cells. Moreover, miR-370 mimics induced cell death of HepG2. Furthermore, Western blotting analysis results demonstrated that miR-370 inhibited the proliferation of liver cancer cells by activating FoxO3a. CONCLUSIONS: MiR-370 inhibited cell proliferation of liver cancer cells by PI3K/Akt signaling pathway. It worked as a tumor suppressor to suppress the progression of human liver cancers.

Application of low tube voltage coronary CT angiography with low-dose iodine contrast agent in patients with a BMI of 26-30 kg/m2.

AIM: To evaluate the feasibility of using a reduced radiation dose and reduced iodine intake (i.e., "double low": low tube voltage/low iodine dose contrast agent) scanning protocol and the adaptive iterative dose reduction (AIDR) reconstruction algorithm in coronary heart disease (CHD) patients with a BMI of 26-30 kg/m(2). MATERIALS AND METHODS: One-hundred and seventy-nine consecutive CHD patients with a body mass index >26 kg/m² but <30 kg/m² were randomly assigned to two groups (group A: 53 men, 39 women, average age 61.83 ± 11.84 years, and group B: 40 men, 47 women, average age 62.25 ± 11.37 years) based on tube voltage, contrast agent, and algorithm used. Group A underwent the "double low" protocol (iodixanol at 270 mg iodine/ml, 100 kVp tube voltage, and AIDR). Group B received the conventional protocol [iopamidol at 370 mg iodine/ml, 120 kVp tube voltage, and filtered back projection (FBP)]. RESULTS: The volume CT dose index (CTDIvol), dose-length product (DLP), effective dose (ED), and iodine intake of patients in "double low" group A were significantly lower than the "conventional" group B (p < 0.001). The mean intraluminal attenuation and contrast enhancement in group A were also significantly less than group B (p < 0.001), whereas the image noise using AIDR in group A was significantly lower than group B using FBP (p < 0.001). However, the signal-to- noise ratio (SNR), contrast-to-noise ratio (CNR), and image-quality scores between the two groups were comparable. CONCLUSIONS: Use of 320-row CT with a "double low" scanning protocol for CCTA in patients with a BMI of 26-30 kg/m(2) not only provided images of diagnostic quality but also reduced both radiation dose and iodine intake during scanning.

Establishing a new electrical conduction pathway by anastomosis of the right auricle and right ventricle assisted by mesenchymal stem cells in a canine model.

BACKGROUND: Electronic pacemakers are the primary treatment of complete atrioventricular (AV) block, but their use is associated with many complications. The aim of the present study was to create an alternative treatment for these patients. MATERIALS AND METHODS: Mesenchymal stem cells (MSCs) isolated from the bone marrow of a 3-month-old dog were cultured in vitro. The MSCs were labeled with 4', 6-diamidino-2-phenylindole (DAPI) before transplantation. We anastomosed the right auricle and right ventricle in 24 dogs, and transplanted labelled MSCs into the anastomotic area of 8 dog hearts. Using immunostaining we assessed survival and differentiation of the implanted cells at 8 weeks posttransplantation. Electrocardiography confirmed the secondary electrical conduction pathway. RESULTS: The ventricular current was captured by the electronic pacemaker in 21 dogs. Compared with the control group (surgery alone), pacemaker stimulus current was significantly less in the MSC group (surgery+MSCs). CONCLUSIONS: Anastomosis of the right auricle and right ventricle assisted by MSCs may be a new treatment for patients with complete AV block in the future.

Eluting characteristics of a platelet glycoprotein receptor antibody using a PLLA-coated stent.

Drug-eluting stents (DES) have emerged as a recognized alternative to treat stent restenosis but many questions remain regarding the optimal type and eluting characteristics of both drug and stent. The first component of the study examines the extent of surface coating of PLLA (poly(L-lactic acid)) on a Nitinol stent. The second characterizes the adsorption and elution rates of monoclonal mouse anti-human platelet glycoprotein (GP) IIIa antibody SZ-21 from a PLLA-coated surface. The PLLA coating was examined by fluorescence staining and image analysis using the Image Processing Box of MATLAB. Stents exposed to the monoclonal mouse anti-human platelet GP IIIa antibody were tested for their adsorption characteristics by radioisotope technique with (125)I-labelled SZ-21. The elution rates were then measured in looped circuits at different velocities (10 or 20 ml/min) and durations (30 min up to 312 h). Results showed that the fluorescence staining and image analysis showed a striking difference in the extent of coating between PLLA-coated stents and SZ-21 eluting stents on the gray-scale distribution of Nitinol surfaces. The amount of SZ-21 adsorbed onto the PLLA-coated stents was dependent on the concentration and duration of immersion in the solution. The method of preparation the mAb eluting stent significantly influenced the elution characteristics for a continuous perfusion of more than 300 h. The eluting curve was biphasic with initial rapid elution for the first 24 h followed by a gradual slow elution. These results indicate that the Image Processing Box of MATLAB appears to be a useful method for semi-quantitative analysis of fluorescence images. Furthermore, SZ-21 can be passively adsorbed onto PLLA-coated stents and predictably influenced by the concentration and duration of immersion. These studies may pave the way to developing stent-based delivery of a potent anti-platelet agent.