PubMed Health⌕ Search

Biomedical subjects

J N Shi

Publications and source records attributed to J N Shi.

9 recordsLinked to original sources

Identification of genetic differences between two clinical isolates of Streptococcus mutans by suppression subtractive hybridization.

Streptococcus mutans is generally considered to be the principal aetiological agent for dental caries. Phenotypic variation in strains is often associated with differences in gene content, so the isolation of DNA fragments from these genes or associated regions is illuminating. The S. mutans strains 9-1 and 9-2, which both colonized the same oral cavity, were selected after screening for the possession of suspected virulence traits. Genomic DNA of strain 9-1 was used as the tester, and that of 9-2 was used as the driver. Suppression subtractive hybridization (SSH) was applied between the tester and the driver DNAs. The subtractive products were cloned into a pCR2.1 vector. Clone libraries representing sequence differences were obtained. The subtractive fragments that were found specifically in strain 9-1 but not in strain 9-2 were identified by dot blotting and then sequenced. BLASTn and BLASTx sequence homology analyses were subsequently performed. Twenty-seven sequences were found in the genome of strain 9-1 that were not in 9-2. Among them, three revealed no homology to published nucleotide sequences while the remaining sequences showed 81-100% homology to known genes of S. mutans strain UA159. These sequences are involved in competence development, signal transduction and transcriptional regulation, repairing stress damage, transport, carbohydrate catabolism, biochemical synthesis, or unknown functions. Differences exist in the genomes of different S. mutans isolates. SSH is effective in screening for S. mutans strain specific DNA sequences.

Adult↗

Evidence that fructose 1,6-bisphosphate specifically protects the alpha-subunit of pyrophosphate-dependent 6-phosphofructo-1-phosphotransferase against proteolytic degradation.

Pyrophosphate-dependent 6-phosphofructo-1-phosphotransferase (PFP) consists of alpha (regulatory) and beta (catalytic) subunits. The alpha-subunit was previously reported to be much more susceptible to tryptic digestion than the beta-subunit. In this study, ligand-induced protection of PFP subunits against proteolysis by subtilisin was investigated in vitro and the data obtained demonstrated that fructose 1,6-bisphosphate (Fru-1,6-P(2)), while exerting negligible effect on the beta-subunit, remarkably protected the alpha-subunit against proteolytic degradation. Western blot analysis revealed a good correlation between the Fru-1,6-P(2) concentration and the degree of corresponding protection on the alpha-subunit against proteolysis. In contrast, none of other examined ligands including fructose 2,6-bisphosphate, fructose 6-phosphate and pyrophosphate had such protection on the alpha-subunit. This finding (1) indicates that the stability of the alpha-subunit can be selectively increased by Fru-1,6-P(2), and (2) suggests that Fru-1,6-P(2) is likely a special effector of the alpha-subunit.

Electrophoresis, Polyacrylamide Gel↗

Enhancing effects of simulated microgravity on Agrobacterium-infected frequency of tobacco callus.

Under the condition of rotation-induced gravity compensation, the time course of interaction between Agrobacteriun tumefaciens and tobacco callus was investigated by means of a scanning electronic microscope. Resulted from repeated experiments, it was found that callus induced from tobacco leaves under simulated microgravity was easier to be infected by A. tumefaciens than controls. Analyses with a scanning electronic microscope indicated that A. tumefaciens were instantly detected on the surface of cell in the first 5 min, that is, A. tumefaciens are liable to interrecognize with callus cell upon contact with each other. With the proceeding of co-culture, the infection efficiency of A. tumefaciens was correspondingly increased. When the time reached 6 h, the fiber was formed between A. tumefaciens and callus cell. In our experiment, the erecting rotating state was taken as the control to exclude the interference of rotating. In this case, A. tumefaciens did not adsorb on calli until 3 h of co-culture, and fiber was only observed as late as 16 h. Statistic data showed that A. tumefaciens-infected frequency of the callus under the action of microgravity was elevated to 176% over that of control.

Agrobacterium tumefaciens↗

Variables affecting electronic root canal measurement.

This study was conducted in two parts. In the first part, 20 single-rooted teeth that had been scheduled for extraction were investigated. The electronic root canal lengths were measured in vivo with a Dental Sono-Explorer type Y-III, and the actual canal lengths were measured after extraction of the teeth. The rate of agreement of the two measurements was 77.5% within a range of +/- 0.5 mm, while it was 100% at +/- 2.0 mm, which is acceptable clinically. In the second part, there were 19 simulated canals whose lengths and apical foramen sizes were known beforehand. Experiments revealed a negative correlation between the areas of the apical foramina and the difference between the electronic and the actual root canal lengths. This relationship was shown by the linear regression equation: ŷ = 0.6-1.6x. With the exception of the smallest areas of foramina, electronic root canal length measurements were less than the actual lengths.

Analysis of Variance↗

Scanning electron microscopic investigation of maxillary lateral incisors with a radicular lingual groove.

Fourteen human lateral incisors with a radicular lingual groove were observed under the scanning electron microscope to explore the communications between the groove and the pulp cavity. Observations showed that accessory foramina could be found not only in the groove, but also in the corresponding wall of the pulp cavity and a cross section. A complete lack of closure of the calcified tissues along the groove, allowing for direct connection of the pulp and the periodontium, was not found in these specimens. Therefore, it is suggested that accessory canals are the main way of communication between the pulp and the periodontium of the incisors with a radicular lingual groove. Because the accessory canals could be found either in the crown part or in the root part of the groove, infectious materials may get into the pulp cavity when there are pathologic conditions either through accessory foramina in the crown part or through accessory foramina exposed by localized periodontitis around the groove.

Dental Pulp Cavity↗

Multilineage differentiation of ectomesenchymal cells isolated from the first branchial arch.

Cranial neural crest-derived ectomesenchymal cells may be pluripotent stem cells that are capable of generating a range of phenotypes. The fate of these cells appears to be determined in part by intrinsic genetic programs and also by the influence of extracellular signals in the local environment. The extent of lineage determination once neural crest cells have migrated to the first branchial arch is not clear, although branchial arch pattern is not thought to be the result of crest predetermination. The aim of the present study was to test the hypothesis that ectomesenchymal cells of the first branchial arch show properties of pluripotent stem cells, the lineage of which may be directed by specific molecular signaling. Ectomesenchymal cells were enzymatically isolated from the mandibular processes of BALB/c mice and maintained in an undifferentiated state while cultured with leukemia inhibitory factor or induced to differentiate by lineage-specific induction factors or growth conditions, including transforming growth factor beta, forskolin, and a mineralization-promoting medium. Morphological observations and immunocytochemistry demonstrated that cells could be induced to differentiate into smooth muscle cells, glial cells, and osteoblasts, respectively. In the presence of the mineralization-promoting medium, alkaline phosphatase activity increased significantly and mineralization nodules formed. The data reported support the concept that many, although not all, first branchial arch-derived ectomesenchymal cells show properties of multipotent stem cells, the subsequent fate of which can be influenced by induction factors and growth conditions. Some cells, however, showed a degree of commitment with respect to their fate. The possible application of first branchial arch-derived stem cells to tissue engineering of the orofacial tissues should involve consideration of the developmental stage of cell harvesting and the desired cell fate.

Animals↗