Autophagy related protein 9A increase in hepatitis B virus-associated hepatocellular carcinoma and the role in apoptosis

The majority of hepatocellular carcinoma (HCC) cases are associated with the hepatitis B virus (HBV) infection. Autophagy related protein 9A (ATG9A) is a transmembrane protein required for autophagosome formation. In order to investigate the role of ATG9A in HBV-associated HCC, ATG9A protein expression was determined in tumor liver tissues and compared with adjacent nontumor tissues from HCC patients with or without HBV infection. In HBV-associated HCC tissues, ATG9A protein level was increased in tumor liver tissues, but not in cases of non-HBV HCC. Our findings suggested that ATG9A might be involved in HBV and cancer cell survival. Therefore, we aimed to analyze the function of ATG9A in HBV replication using RNA interference to evaluate the HBV DNA level using real-time PCR. In the present study, there were no significant differences between shATG9A-transfected HepG2.2.15 cells and the mock control. However, we found that silencing ATG9A affected apoptosis in HepG2.2.15 and HepG2 cell lines. Our results indicated that ATG9A might be partly involved in the survival of HCC. Thus, the inhibition of ATG9A together with other targets might be a potential drug target for HCC treatment.     

Validation of osmotic fragility test and dichlorophenol indophenol precipitation test for screening of thalassemia and Hb E

The strategy for screening of thalassemia and Hb E by a combination of osmotic fragility (OF) test and dichlorophenol indophenol precipitation (DCIP) test was validated with 436 unrelated Thai subjects. Hemoglobin (Hb) typing, Hb A2 quantitation, PCR and DNA sequence analysis were used as confirmatory methods for diagnosis of thalassemia and hemoglobinopathy. The sensitivity and specificity of this strategy was 100% and 79.7%, respectively. The results assessed by two medical scientists were exactly the same with 93.3% accuracy in comparison with the confirmatory methods. A combination of OF and DCIP has been shown to be a reliable, rapid, simple and sensitive strategy for screening thalassemia and Hb E in the Thai population.     

Local botulinum toxin type A injections in the treatment of spastic toes

OBJECTIVE: To investigate the efficacy and safety of botulinum toxin type A treatment of spastic toes using varying doses based on the degree of spasticity (Modified Ashworth Scale). DESIGN: Single-center, open-label, prospective study. Hemiplegic patients with either hitchhiker's great toes (persistent extension of the great toes) or toe flexor spasms with pain during walking were treated with local intramuscular injections of botulinum toxin type A. Initial botulinum toxin type A dose per muscle was 25 units for patients with a baseline Ashworth score of 2, 50 units for a score of 3, and 75 units for a score of 4. Additional botulinum toxin type A injections were allowed if there was an insufficient clinical response to initial treatment. The muscles injected included flexor digitorum, extensor hallucis longus, and/or flexor hallucis longus. All injections were made using electromyographic guidance. Outcome measures were the Modified Ashworth Scale, a visual pain scale, a visual percentage of function scale, and adverse effects. RESULTS: Twenty patients were enrolled. The dose of botulinum toxin type A used ranged from 25 to 35 units per muscle for an Ashworth score of 2, from 50 to 70 units per muscle for a score of 3, and from 75 to 95 units per muscle for a score of 4. There were improvements in all outcome measures. In most patients, the benefits lasted 5-6 mo, with a few patients exhibiting benefits for > or =2 yr. There were no adverse effects. CONCLUSIONS: Botulinum toxin type A treatment using doses based on spasticity severity seems to be safe and effective in the treatment of spastic toes, and further study is warranted.     

Autoxidation of brain homogenates from various animals as measured by thiobarbituric acid assay

INTRODUCTION: The TBARS assay has been well recognized for determination of lipid peroxidation and oxidative injury in biological samples including brain homogenates. In general, the homogenates are freshly prepared using rat brains as the tissue sources. In this study, we compared the rates of spontaneous lipid peroxidation in brain homogenates obtained from bovine, canine, hen, rat, and swine. In addition, the influences of lyophilization process and storage time up to six months at -20 degrees C without the freeze-thaw cycle were also determined in the swine brain preparations. METHODS: The standard assay for thiobarbituric acid-reactive substances (TBARS) was performed at 37 degrees C, using spectrophotometry to quantify the malondialdehyde (MDA) content. RESULTS: Rat brain homogenate exhibited the highest autoxidation rate (0.128+/-0.002 microM/min) whereas the bovine brain exhibited the lowest rate (0.032+/-0.001 microM/min). Swine brain homogenate could be kept at -20 degrees C up to 3 months without a significant increase in rate of autoxidation. Lyophilization caused a significant increase in the autoxidation rate of brain homogenate. However, the autoxidation rates of the lyophilized preparation were quite comparable throughout the six-month freezing time. DISCUSSION: Swine brain was a good candidate for tissue source in the TBARS reaction. The homogenate could be kept in the lyophilized form under the storage condition at -20 degrees C without the freeze-thaw cycle in the dark for at least six months.     

Constituents of Polyalthia jucunda. and Their Cytotoxic Effect on Human Cancer Cell Lines

Abstract

4-Hydroxy-4, 7-dimethyl-α.-tetralone (1), 4,5-dihydroblumenol A (2), N.-trans.-feruloyltyramine (3), and 24–methylenelanosta-7, 9(11)-dien-3-β., 15α.-diol (4) were isolated from the stem bark of Polyalthia jucunda. (Piere) Finet & Gagnep (Annonaceae). All the compounds were evaluated for their effects on growth of four human tumor cell lines [ER(+) MCF-7, ER(?) MDA-MB-23, SF 268, and NCI-H460] and of a non–tumor cell line (MRC-5). Only compound 4 exhibited a dose-dependent growth inhibitory effect against both tumor and non–tumor cell lines but with less effect on the latter. Using the TUNEL assay, it was found that the inhibitory effect of compound 4 on NCI-H460 cells was probably caused by apoptosis.     

Pulpal anesthesia in pediatric patients following supplemental mandibular buccal infiltration in vital permanent mandibular molars with deep caries

Clinical Oral Investigations - Inferior alveolar nerve block (IANB) does not always provide adequate pulpal anesthesia, and supplemental techniques have been investigated in adults. This study...     

Delta-like ligand 4 in hepatocellular carcinoma intrinsically promotes tumour growth and suppresses hepatitis B virus replication

AIM To investigate the role of Delta-like ligand 4(DLL4) on tumour growth in hepatitis B virus(HBV)-associated hepatocellular carcinoma(HCC) in vivo.METHODS We suppressed DLL4 expression in an HBV expressing HCC cell line, HepG2.2.15 and analysed the growth ability of cells as subcutaneous tumours in nude mice. The expression of tumour angiogenesis regulators, VEGF-A and VEGF-R2 in tumour xenografts were examined by western blotting. The tumour proliferation and neovasculature were examined by immunohistochemistry. The viral replication and viral protein expression were measured by quantitative PCR and western blotting, respectively.RESULTS Eighteen days after implantation, tumour volume in mice implanted with sh DLL4 HepG2.2.15 was significantly smaller than in mice implanted with control HepG2.2.15(P 0.0001). The levels of angiogenesis regulators, VEGF-A and VEGF-R2 were significantly decreased in implanted tumours with suppressed DLL4 compared with the control group(P 0.001 and P 0.05, respectively). Furthermore, the suppression of DLL4 expression in tumour cells reduced cell proliferation and the formation of new blood vessels in tumours. Unexpectedly, increased viral replication was observed after suppression of DLL4 in the tumours.CONCLUSION This study demonstrates that DLL4 is important in regulating the tumour growth of HBV-associated HCC as well as the neovascularization and suppression of HBV replication.     

Outcomes of Direct Pulp Capping by Using Either ProRoot Mineral Trioxide Aggregate or Biodentine in Permanent Teeth with Carious Pulp Exposure in 6- to 18-Year-Old Patients: A Randomized Controlled Trial

Introduction

This study aimed to compare the success rates of direct pulp capping (DPC) by using either ProRoot Mineral Trioxide Aggregate (MTA) or Biodentine in the cariously exposed permanent teeth of 6- to 18-year-old patients. Gray discoloration was also evaluated.