Natural compound fraxinellone ameliorates intestinal fibrosis in mice via direct intervention of HSP47-collagen interaction in the epithelium.

Intestinal fibrosis is a common complication of inflammatory bowel disease. There is still a lack of effective drugs for the prevention or treatment of intestinal fibrosis. Heat shock protein 47 (HSP47) plays a key role in the development of intestinal fibrosis. In this study we investigated the therapeutic potential and underlying mechanisms of fraxinellone, a degraded limonoid isolated from the root bark of Dictamnus dasycarpus, in the treatment of intestinal fibrosis. Intestinal fibrosis was induced in mice by dextran sodium sulfate (DSS) treatment. DDS-treated mice were administered fraxinellone (7.5, 15, 30 mg·kg-1·d-1, i.g.) for 45 days. We showed that fraxinellone administration dose-dependently alleviated DSS-induced intestinal impairments, and reduced the production of intestinal fibrosis biomarkers such as α-smooth muscle actin (SMA), collagen I, hydroxyproline, fibronectin and laminin, and cytokines such as TGF-ß, TNF-α and IL-ß. We then established in vitro intestinal fibrosis cell models in SW480 and HT-29 cells, and demonstrated that treatment with fraxinellone (3, 10, 30 µM) significantly relieved TGF-ß-induced fibrosis responses by inhibiting the TGF-ß/Smad2/3 signaling pathway. Molecular docking suggested that the fraxinellone might disrupt the interaction between HSP47 and collagen, which was confirmed by coimmunoprecipitation experiments. SPR analysis showed that fraxinellone had a high affinity for HSP47 with a Kd value of 3.542 × 10-5 M. This study provides a new example of HSP47-collagen intervention by a natural compound and has important implications for the clinical treatment of inflammation-induced issue fibrosis.

Clinical analysis on the root fracture of the maxillary first molar.

OBJECTIVES: This study aimed to investigate the common types and directions of root fractures of the maxillary first molar and the influence of root canal treatment on the prevalent sites of root fractures. METHODS: A total of 274 maxillary first molars with root fractures diagnosed via cone beam computed tomography were included. The root fractures of nonendodontically and endodontically treated teeth were identified to be spontaneous and secondary root fractures, respectively. The sites, types, and directions of spontaneous and secondary root fractures were determined. RESULTS: Among the spontaneous root fractures, the proportion of palatal root fractures (56.1%) was higher than those of mesial buccal root fractures (36.1%) and distal buccal root fractures (7.8%). Among the secondary root fractures, the proportion of mesial buccal root fractures (52.7%) was higher than those of palatal root fractures (36.5%) and distal buccal root fractures (10.8%). The distribution of predominant fracture sites was statistically significant (P<0.05), and vertical root fracture was the most common type. Palatal and buccal roots were commonly fractured at the mesiodistal and buccal-palatal directions, respectively. CONCLUSIONS: This study provided an epidemiological basis for the clinical features of root fractures of the maxillary first molar. During the dia-gnosis and treatment of the maxillary first molar, the possibility of palatal root fractures should be considered. The occurrence of mesial buccal root fractures may be related to root canal treatment. Therefore, the risk of mesial buccal root fractures caused by iatrogenic factors should be minimized.

cDNA cloning, difference expression in various organs, and bioinformatic analysis of HMGS gene in Sambucus chinensis / 中草药

Objective: To clone 3-hydroxy-3-methylglutaryl coenzyme A synthetase (HMGS) gene from Sambucus chinensis and analyze the difference expression. Methods: The sequence of HMGS gene was cloned from S. chinensis by using RT-PCR strategy. The physiochemical properties, secondary structure, and three-dimensional structure of HMGS protein were forecasted and analyzed, and its structure and function were predicted. And the difference expression of HMGS gene in the rhizome, stems, leaves, and flowers of S. chinensis was analyzed by fluorescent quantitative PCR. Results: The cDNA contains a 1 401 bp open reading frame and encodes a predicted protein of 466 amino acids. No transmembrane region and no signal peptide were present in HMGS protein. Relative real-time PCR analysis indicated that HMGS gene showed the higher transcript abundance was in the flowers and rhizomes, while was lower in the leaves. Conclusion: The HMGS gene is first cloned from S. chinensis and the result will provide a foundation for elucidating the mechanism of the gene in the metablism pathway of terpenoid in S. chinensis.

Cloning, expression, and bioinformatics analysis of acetyl-CoA C-acetyltransferase gene in Houttuynia cordata / 中草药

Objective: Acetyl-CoA C-acetyltransferase (AACT) is the initial enzyme in the terpenoid biosynthesis pathway of mevalonate (MVA), two units of acetyl-CoA were catalyzed to acetoacetyl-CoA. To clone the full length cDNA of AACT gene and carry out the bioinformatics analysis and expression analysis in order to provide the basis on resolving the mechanism of biosynthesis for terpenoid secondary metabolites from Houttuynia cordata. Methods: The cDNA sequence of AACT gene was obtained from H. cordata by using RT-PCR strategy. And the different expression of AACT gene in the rhizomes, stems, leaves, and flowers of H. cordata was analyzed by fluorescent quantitative PCR. Results: The cDNA contains a 1 218 bp open reading frame and encodes a predicted protein of 405 amino acids. No transmembrane region and signal peptide were present in AACT protein by bioinformatics prediction. Relative real-time PCR analysis indicated that AACT gene showed the highest transcript abundance in the stems and rhizomes of H. cordata lower levels in the flowers and leaves, the values of them were 1.49, 0.96, 0.20, and 0.10, respectively. Conclusion: This AACT gene is cloned from H. cordata for the first time. The results will provide a foundation for exploring the mechanism of the gene in terpenoid biosynthesis and metabolism in H. cordata.

Molecular cloning, differential expression, and bioinformatics analysis of HMGR gene in Sambucus chinensis / 中草药

Objective: To clone the 3-hydroxy-3-methylglutaryl coenzyme A redutase (HMGR) gene from Sambucus chinensis and analyze the differential expression. Methods: The sequence of HMGR was cloned from S. chinensis by using RT-PCR strategy. The physical and chemical properties, secondary structure, and tertiary structure of the HMGR protein were forecasted and analyzed, and its structure and function were predicted. And the different expression of HMGR gene in the rhizome, stems, leaves, and flowers was analyzed by fluorescent quantitative PCR. Results: The cDNA contains a 1782 bp open reading frame and encodes a predicted protein of 593 amino acids. Two transmembrane regions and no signal peptide were present in HMGR. Relative real-time PCR analysis indicated that HMGR showed the higher transcript abundance in the flowers and aerial stems, and the lower levels in the rhizomes and leaves. Conclusion: This study clones and expression analyzes HMGR gene from S. chinensis for the first time. The results will provide a foundation for exploring the mechanism of terpenoid biosynthesis in S. chinensis.

Chemical constituents of stems of viola japonica var. stenopetala Franch. ex H / 中国药学杂志

OBJECTIVE: To investigate the chemical constituents from the stems of Viola japonica var. stenopetala Franch. ex H. METHODS: The constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography, and preparative TLC. The structures were identified on the basis of spectral data and physiochemical characteristics. RESULTS: Fifteen compounds were isolated from 70% ethanol extract of Viola japonica var. stenopetala Franch. ex H. and identified as β-sitosterol (1), daucosterol (2), chlorogenic acid (3), 7-hydroxycoumarin (4), stigmastero-3-O-β-D-glucopyranoside (5), dehydrololiolide (6), kaempferol-7-O-β-D-glucopyranoside (7), characterizedas(+)-pinoresinol-O-β-D-glucopyranoside (8), 5, 7-dihydroxy-3, 6-dimethoxyflavone (9), apigenin-7-O-β-D-glucoside (10), chryseriol (11), β-amyrin (12), robinin (13), kaempferol-3, 7-di-O-α-L-rahmnoside(14), and solagenin-6-O-β-D-quinovopyranoside(15). CONCLUSION: Compounds 8 and 15 are isolated from the plants in Gnaphalium L. for the first time. Compounds 5, 6, 8, 11, 14, and 15 are isolated from this plant material for the first time.

[An anatomical study of maxillary sinus septum of Han population in Jiangsu region using cone-beam CT].

PURPOSE: To examine the anatomical variation of maxillary sinus septum of Han nationality in Jiangsu region by using cone-beam computed tomography (CBCT) combined with Simplant software in order to provide anatomical basis and operation instruction for oral implants after maxillary sinus lifting. METHODS: CBCT image data were collected from 424 patients for analysis of maxillary sinus septa. Digital imaging and communications in medicine (Dicom) image files were fed into the computer-aided Simplant software and used to analyze the prevalence, location, height, orientation, and morphology of maxillary sinus septa through three-dimensional reconstruction. The data was analyzed with SPSS17.0 software package. RESULTS: The proportion of the occurrence of maxillary sinus septa in 424 subjects was 44.81% and 21.23% of the subjects (n=90) had multiple sinus septa, while 20.52% had bilateral sinus septa (n=87). Totally 848 maxillary sinuses were observed in this study and 277 sinuses had septa with a proportion of 32.67%. The prevalence of septa was not significantly related to gender, age, and the presence or absence of teeth. Septa were located most frequently in the middle of maxillary sinus (59.94%). The mean height of sinus septa was (5.90±3.65) mm and (5.54±2.87) mm in the right and left maxillary sinus, respectively. The mean length of sinus septa was (8.15±2.40) mm and (7.88±2.73) mm in the right and left maxillary sinus, respectively. CONCLUSIONS: Nearly 44.81% of Han population in Jiangsu region have maxillary sinus septa. The CBCT imaging technique can provide comprehensive and accurate quantitative analysis of maxillary sinus septa and is meaningful to provide anatomical basis and clinical guidance before sinus augmentation procedures.