Many methods have been developed for more efficient gene delivery and expression in human cells. A number of studies have been performed in achieving successful gene delivery and expression conditions. We investigated differential gene expression patterns after delivery adenoviral vector containing green fluorescent protein(GFP) gene into human cancer cell lines. We constructed recombinant adenoviral Ad-CMV-GFP containing CMV promoter and GFP gene. The efficiency of gene expression was assessed by observation GFP expressing cells using fluorescent microscopy after transfer of Ad-CMV-GFP in concentrations of 0.1μl. 1μl. 10μl. At first, we evaluated expression patterns of gene in several human cancer cell lines, gastric adenocarcinoma cell line AGS was showed high level of GFP expression compared with colorectal adenocarcinoma cell line HT-29. After transfer 0.1μl of Ad-CMV-GFP in AGS, we could found that GFP expression cells were observed in next day and highly increased 2 days. While, small number of GFP expressing cells were examined in HT-29 and SNU-C4. Therefore, these data showed that AGS was expressed the highest level of GFP and almost AGS cells seems to express GFP in concentration of 1μl of Ad-CMV-GFP. GFP expression pattern in HT-29 reveal that expression was low in next day after gene transfer but significantly increase expression level in 2 days. In case of SNU-C4, GFP expression increased with increasing concentration of Ad-CMV-GFP and t ransfer times. For examine effects of transfer times in small amount gene, we transfer in concentration of 0.1μl Ad-CMV-GFP and detected GFP expression patterns after 2 days or 4 days. As a result, expression level of GFP in AGS was increase about 2 fold after 4 days compared with 2 days, but any difference of GFP expression levels were not showed in HT-29 and SNU-C4. Our study suggested that adenovirus was very efficient gene transfer vector for gene expression in human cancer cell lines. In addition to, we also demonstrated that gene expression patterns was dependent on each human cell lines. Therefore, further studies will be needed to confirm the optimum conditions for efficient gene delivery and expression in each target cell lines with consideration to cellular properties.
The regeneration of periodontium is the goal of periodontal therapy. Many periodontologists try to achieve this goal by using guided tissue regeneration(GTR) method. However, these procedures always include several disadvantages. Recombinant human bone morphogenetic protein-2 (rhBMP-2) stimulated ectopic bone formation when it was implanted in rat muscles with insoluble bone matrix by differentiating muscle cells into chondrocytes and osteoblasts. The purpose of this study was to evaluate the osteoinductive potential of the mixture of rhBMP-2 (5 μg/ml) and heparin (0.25 or 25 μg/ml ) at the critically sized rabbit calvarial defects. And this study aimed to reveal that heparin also acts to enhance the bone forming activity of rhBMP-2. The 12 rabbits (4-month-old; NewZealand White) were used in the present study. 5 μg/ml of rhBMP-2 and 0, 0.25 or 25 μg/ml of heparin were mixed and blotted into anorganic bovine bone and filled cranial defects. The animals were sacrificed following a time schedule (1, 3, and 6 weeks). Sections were made in 7 μm thicknesses, stained with H&E and Masson's trichrome method, and examined under a light microscope. The differences among each obtained percent value were evaluated by one-way analysis of variance. A p value of p<0.05 was considered statistically significant and an ANOVA test was performed to verify significant differences. To adjust for multiple comparisons when one-way analysis of variance showed a significant difference between groups (p<0.05), Scheffe`s post hoc test was used to identify which group differences accounted for the significant p-value. In control group, osteoinduction was not outstanding, however, in experimental groups, osteoinduction was significantly outstanding, and as the concentration of heparin mixed with rhBMP-2 increased, osteoinduction was increased. Mixtures of rhBMP-2 and heparin affect bone formation at initial bone formation, but that effect disappeared following a time lapse.
In the previous molecular cloning study from human salivary gland cDNA l ibrary a novel clone (C77-091) was known as a candidate gene for antimicrobial protein by GenBank database search and RNA in situ hybridization. This study is aimed to identify the molecular characteristics of C77-091 protein, which showed an antimicrobial activity on E.coli, thereby named as salivary antimicrobial protein (SAMP). SAMP consisted of a typical hydrophobic amino acid rich domain in the N-terminus, a cluster of basic amino acids, carbohydrate attachment site, a possible transglutaminase catalyzed cross-linking site, and multiple consensus sequences of phosphorylation site in the C-terminus. Western blot analysis of human organs and tissue with the monospecific antibody to the synthetic SAMP peptide showed strong interacting protein from the extracts from submandibular gland and parotid saliva but absent in the mixed saliva, and the immunohistochemical staining detected a strong positive regions in the secretory granules in the luminal cytoplasm of interlobular ductal cells of salivary gland. The SAMP was also distributed in the human sebaceous gland and prostate. These data suggest that C77-091 named SAMP gene is a novel antimicrobial protein in human salivary gland, which may play a role for the innate immunity by protecting and stabilizing the mucosal epithelium to maintain homeostasis of oral mucosa.
Halitosis is a major problem on social interactions in many people, and oral hygiene control is a major treatment for halitosis. However, medication may be the choice of method for non-oral origin halitosis. The present study evaluated the efficacy of chlorophyllin medication for oral malodor patients with GERD (gastroesophageal reflux disease). 26 patients with halitosis visited Department of Periodontology of Seoul National University Bundang Hospital. VSCs (Volatile Sulfur compounds) level was evaluated according to Chlorophyllin medication after 20 days’ medication. Scaling was conducted after medication, and VSCs level was measured 28 days after scaling. Chlorophyllin medication on the halitosis patients with GERD showed a statistically significant efficacy (P > 0.05). There was no significant effect on the patients without GERD (P>0.05). There was no significant effect s caling before o ne month ( P>0.05). Chlorophyllin was e ffective in t he halitosis patients with GERD. There was no significant difference one month after scaling.
A 37 years old male patient had suffered from fibrous nasal polyp for more than 10 years even though careful medical treatment, and recently aggravated with severe nasal stiffness and pain. In the radiological observation the lesion was continuously extended into the ipsilateral maxillary sinus by poly cystic radiopacities gradually destroying the anterior wall of antrum. During the Caldwell-Luc operation a cystic fibrous granulomatous tissue was removed, and its microsections showed multicystic and tunnel-like spaces, of which lumens were usually round and filled with parts of parasitic worm e xhibiting parasitic t eguments and eggs. The granulomatous l esion showed relatively mild inflammatory reaction with the infiltration of eosinophilic PMNs. The histological features of tunnel-like penetration of parasitic worms, and their teguments and eggs were competent with the diagnosis of sparganosis. Especially, although the patient had no experience of raw foods such as frog or snake, he used to drink the untreated natural stream water obtained in the mountain area of Gangwon prefecture in Korea. Therefore, the present study demonstrated a rare lesion of antral spanganosis, and we suppose that the frequent incident of oral sparganosis in Gangwon prefecture is possibly related to the usage of untreated stream water in the mountain area of Gangwon prefecture in Korea
Facial numbness restricted to the distribution of the mental nerve(mental neuropathy) is called "numb chin syndrome". The clinical importance of this syndrome is associated with first recognition of involvement of malignant diseases. The malignant neoplasm with numb chin syndrome show rapid progression and high mortality. We present a 43-year-old female who had been treated by radiotherapy for precursor T-cell leukemia/lymphoma involving the central nervous system( CNS) previously and later developed mental nerve invasion without central nervous system recurrence. MRI images revealed the CNS tumor mass remitted, and there was no identified peripheral nervous system(PNS) involvement including the mental nerve invasion, nevertheless the patient complained of consistent numbness and pain on right mandibular area. This is the first case of precursor T-cell leukemia/lymphoma accompanying mental nerve invasion without recurrence for central nervous system. Proper interpretation for mental neuropathy may lead to the prompt diagnosis and therapeutic intervention.
Epithelioid hemangioma is an unusual, which was at first described as angiolymphoid hyperplasia with eosinophilia (ALHE) in 1969 and misinterpreted as the same disease with Kimura’s disease. But now it represents a distinctive vascular tumor, a different entity with those inflammatory conditions. Here, we present a case of epithelioid hemangioma appearing as a diffuse gingival nodular mass in 33‐year‐old male who had suffered from the idiopathic sharp pain of left mandible area previously and later extended to left maxillary region, therefore the gingival lesion was clinically estimated as malignancy associated with neural invasion. CT images revealed the soft tissue mass restricted to left maxillary gingiva without infiltration to underlying maxillary alveolar bone. Histologically, the circumscribed soft tissue mass consist of solid sheets of epithelioid to spindled cells, expressing reactivity for neither cytokeratin AE1/3 nor HMB‐45, but for CD31, endothelial cell marker. Therefore, the lesion was pathologically diagnosed as epithelioid hemangioma. Diagnostic evaluation for the neurologic symptoms of the patient revealed the trigeminal neuralgia, accompanying with the present epithelioid hemangioma incidentally. Awareness of epithelioid hemangioma should be emphasized not to be misdiagnosed as malignant disease like an invasive squamous cell carcinoma, malignant melanoma, or angiosarcoma. Proper evaluation and interpretation for neural symptom may lead to the correct differential diagnosis and therapeutic intervention