[Comparison of biological characteristics of human gingival junctional epithelial cells and oral epithelial cells].

OBJECTIVE: To isolate P-cadherin positive and negative oral gingival epithelial cells, and to compare the biological characteristics with junctional epithelial cells. METHODS: Human oral gingival epithelial cells and junctional epithelial cells were cultured. P-cadherin positive and negative cells were isolated from oral gingival epithelial cells. The cellular adhesion, proliferation and migration were measured and compared. RESULTS: The P-cadherin positive cells accounted for 20% of oral gingival epithelial cells. Compared with juctional epithelial cells, P-cadherin positive oral gingival epithelial cells showed similar properties of adhesion and migration, and stronger proliferation ability (0.72 ± 0.06 vs. 0.60 ± 0.05, P<0.05). P-cadherin negative oral gingival epithelial cells showed weaker ability of adhesion (48% ± 6% vs. 87% ± 11%, P<0.05), proliferation (0.36 ± 0.04 vs. 0.60 ± 0.05, P<0.05) and migration (10.3 ± 2.7 vs. 23.4 ± 4.8, P<0.05). CONCLUSION: P-cadherin positive oral gingival epithelial cells showed some similar but different biological characteristics, compared with juctional epithelial cells, which suggested that during the process of transforming oral gingival epithelial cells into juctional epithelial cells, complex gene and protein changes were involved instead of simply cellular migration.

Integration of a Cas12a-mediated DNAzyme actuator with efficient RNA extraction for ultrasensitive colorimetric detection of viral RNA.

Developing highly sensitive and specific on-site tests is imperative to strengthen preparedness against future emerging infectious diseases. Here, we describe the construction of a Cas12a-mediated DNAzyme actuator capable of converting the recognition of a specific DNA sequence into an amplified colorimetric signal. To address viral RNA extraction challenges for on-site applications, we developed a rapid and efficient method capable of lysing the viral particles, preserving the released viral RNA, and concentrating the viral RNA. Integration of the DNAzyme actuator with the viral RNA extraction method and loop-mediated isothermal amplification enables a streamlined colorimetric assay for highly sensitive colorimetric detection of respiratory RNA viruses in gargle and saliva. This assay can detect as few as 83 viral particles/100 µL in gargle and 166 viral particles/100 µL in saliva. The entire assay, from sample processing to visual detection, was completed within 1 h at a single controlled temperature. We validated the assay by detecting SARS-CoV-2 in 207 gargle and saliva samples, achieving a clinical sensitivity of 96.3 % and specificity of 100%. The assay is adaptable for detecting specific nucleic acid sequences in other pathogens and is suitable for resource-limited settings.

Chromium on the hands of children after playing in playgrounds built from chromated copper arsenate (CCA)-treated wood.

Children's exposure to arsenic and chromium from playground equipment constructed with chromated copper arsenate (CCA)-treated wood is a potential concern because of children's hand-to-mouth activity. However, there exists no direct measure of Cr levels on the hands of children after playing in such playgrounds. In this study we measured both soluble and total Cr on the hands of 139 children playing in playgrounds, eight of which were constructed with CCA-treated wood and eight of which were not. Children's age and duration of play were recorded. The hands of each child were washed after play with 150 mL deionized water, which was collected in a bag and subsequently underwent analysis of Cr and 20 other elements, using inductively coupled plasma mass spectrometry. Total average Cr on the hands of 63 children who played in CCA playgrounds was 1,112 +/- 1,089 ng (median, 688; range 78-5,875). Total average Cr on the hands of 64 children who played in non-CCA playgrounds was 652 +/- 586 ng (median, 492; range 61-3,377). The difference between the two groups is statistically significant (p < 0.01). Cr levels were highly correlated to both Cu (r = 0.672) and As (r = 0.736) levels in CCA playgrounds (p < or = 0.01), but not non-CCA playgrounds (r = 0.252 and 0.486 for Cu and As, respectively). Principal-component analysis indicates that Cr, Cu, and As are more closely grouped together in CCA than in non-CCA playgrounds. These results suggest that the elevated levels of Cr and As on children's hands are due to direct contact with CCA wood.

[Study on the pretreatment of the series of HPD macroporous adsorption resin].

OBJECTIVE: To develop a method for the pretreatment of the series of HPD macroporous adsorption resin (MAR). METHOD: The effect of 2% NaOH to pretreatment of the series of HPD MAR was studied by gas chromatography and UV spectrometery. RESULT: The eluting effect by ethanol was better after the MAR was marinated and eluted by 2% NaOH. CONCLUSION: The pretreatment of the series of HPD MAR was: after marinated and eluted by 2% NaOH, the series of HPD MAR were eluted by ethanol at 2 BV x h(-1) at 60 degrees C for 3-4 bed volumes (BV). At then the UV absorbance of eluate was 0.2-0.5. Benzene was not detected and the other residues less than 10 mg x L(-1) in eluted-MAR by GC. And the MARs match to the standard of medicine.

Kindler syndrome protein Kindlin-1 is mainly expressed in adult tissues originating from ectoderm/endoderm.

Mutations of integrin-interacting protein Kindlin-1 cause Kindler syndrome and deregulation of Kindlin-1 is implicated in human cancers. The Kindlin-1-related diseases are confined in limited tissue types. However, Kindlin-1 tissue distribution and the dogma that governs Kindlin-1 expression in normal human body are elusive. This study examined Kindlin-1 expression in normal human adult organs, human and mouse embryonic organs by immunohistochemical analyses. We identified a general principle that the level of Kindlin-1 expression in tissues is tightly correlated with the corresponding germ layers from which these tissues originate. We compared the expression of Kindlin-1 with Kindlin-2 and found that Kindlin-1 is highly expressed in epithelial tissues derived from ectoderm and endoderm, whereas Kindlin-2 is mainly expressed in mesoderm-derived tissues. Likewise, Kindlin-1 was also found highly expressed in endoderm/ectoderm-derived tissues in human and mouse embryos. Our findings indicate that Kindlin-1 may play an importance role in the development of endoderm/ectoderm related tissues.

Enhanced osteogenic differentiation of mesenchymal stem cells on poly(L-lactide) nanofibrous scaffolds containing carbon nanomaterials.

Carbon nanomaterials (CNMs), such as carbon nanotube (CNT) and graphene, are highlighted in bone regeneration because of their osteoinductive properties. Their combinations with nanofibrous polymeric scaffolds, which mimic the morphology of natural extracellular matrix of bone, arouse keen interest in bone tissue engineering. To this end, CNM were incorporated into nanofibrous poly(L-lactic acid) scaffolds by thermal-induced phase separation. The CNM-containing composite nanofibrous scaffolds were biologically evaluated by both in vitro co-culture of bone mesenchymal stem cells (BMSCs) and in vivo implantation. The nanofibrous structure itself demonstrated significant enhancement in cell adhesion, proliferation and oseogenic differentiation of BMSCs, and with the incorporation of CNM, the composite nanofibrous scaffolds further promoted osteogenic differentiation of BMSCs significantly. Between the two CNMs, graphene showed stronger effect in promoting osteogenic differentiation of BMSCs than CNT. The results of in vivo experiments revealed that the composite nanofibrous scaffolds had both good biocompatibility and strong ability in inducing osteogenesis. CNMs could remarkably enhance the expression of osteogenesis-related proteins as well as the formation of type I collagen. Similarly, the graphene-containing composite nanofibrous scaffolds demonstrated the strongest effect on inducing osteogenesis in vivo. These findings demonstrated that CNM-containing composite nanofibrous scaffolds were obviously more efficient in promoting osteogenesis than pure polymeric scaffolds.

Mechanical properties of hydroxyapatite whisker-reinforced bis-GMA-based resin composites.

OBJECTIVE: To evaluate the reinforcement efficacy of hydroxyapatite (HA) whiskers in bis-GMA-based dental restorative composites and determine the effect of volume fraction on the mechanical properties. METHODS: Silanized HA whiskers and nano-scale powder were mixed in various proportions with bis-phenol A-glycidyl methacrylate (bis-GMA)-based polymer pastes. Equal parts of initiator and accelerator pastes were then mixed by hand. After curing at 25 ± 2 °C and storage in distilled water at 37 °C for 24h, elastic modulus, fracture strength and work-to-failure in three-point bending, fracture toughness using a notchless triangular prism fracture method, and Vickers hardness were determined. Data were examined by means of one-way analysis of variance and linear regression. RESULTS: Reinforcing efficacy was significantly dependent on filler morphology. Whiskers had good dispersibility and wettability with bis-GMA-based polymer, conferring good reinforcement and toughening, significantly better than did the HA nano-scale powder. SIGNIFICANCE: HA whiskers provided better mechanical properties in bis-GMA-based composites compared with the nano-scale powder. Such whisker-reinforced materials may be beneficial compared with currently used dental restorative materials.

Failure and behavior in water of hydroxyapatite whisker-reinforced bis-GMA-based resin composites.

Failure mode under Hertzian indentation and the behavior on immersion in water of bis-GMA-based composites with HA whiskers or nanoscopic HA powder as filler were evaluated. Failure load decreased with increase in filler loading, but the decrease was smaller for whiskers, which showed a different failure mode both macroscopically and microscopically. Particle-filled composites failed mainly by radial cracking and cone cracking, with some plastic deformation at low filler loading, with fracture into irregular segments. For whisker-filled materials, crack propagation was inhibited by the well-dispersed whiskers by the usual toughening mechanisms; cone cracking was the dominant failure mode, at higher loads than for the powder, and fracture was incomplete. The filler reduced both water-uptake and elution of soluble materials, as expected, but both were lower for the whisker-filled material. Such composites might form the basis of viable materials for dental load-bearing restorations and other applications.

Effect of surface treatment of hydroxyapatite whiskers on the mechanical properties of bis-GMA-based composites.

The mechanical properties of bis-GMA-based composites filled with hydroxyapatite (HA) particles or whiskers are characterized in this paper. The inherent properties of reinforcements, the bonding strength of the filler-matrix interface and their dispersibility in a polymer matrix were intimately associated with the mechanical performance of the composites. Silanization of both particles and whiskers effectively improved the bonding of the filler-matrix interface, their dispersibility in matrix monomers and filler loading. The particle- and whisker-filled composites showed highly significant differences in both flexural strength and fracture toughness. HA whiskers prepared by hydrothermal homogeneous precipitation had superior dispersibility and wettability in the polymer matrix; no aggregation and entanglement were found in both the products before and after silanization and the resin composites. These features conferred the whiskers having good interface bonding with the polymer matrix and superior reinforcing and toughening effects. For the particulate filler, the addition of HA led to a decrease in the flexural strength for both silane-treated and untreated fillers. Cracks propagated easily through the micropores and voids in the HA aggregates, leading to lower flexural strength and low toughness. However, silanization of HA did not show obvious effects on the elastic modulus of the composites.

[Preparation of anti-P21-activated kinase 5 polyclonal antibody and its application in dental germ cells].

OBJECTIVE: To clone PAK5-N terminal sequence for expression in E. coli to prepare its polyclonal antibody, and examine the role of PAK5 in dental germ cells. METHODS: Based on human PAK5 cDNA sequence, PCR primers were designed to amplify PAK5-N terminal sequence. The PCR product was cloned into the expression vector pGEX-4T-1 EcoRI/XhoI sites, and the recombinant plasmids were identified by agarose gel electrophoresis followed by DNA sequence analysis. The recombinant plasmids were transformed into E. coli BL21 and the expression of GST-fusion protein was induced by IPTG. Glutathione-Sepharose beads were used to purify GST-fusion PAK5-N-terminal fragment. Anti-PAK5 polyclonal antibody was obtained in immunizing rabbits with purified GST-PAK5 N-terminal fusion protein, and the antibodies were purified by protein A beads and used for detection of PAK5 expression in dental germ cells. RESULTS AND CONCLUSIONS: We successfully cloned PAK5-N terminal gene fragment, and achieved protein expression, purification and production of PAK5-NT polyclonal antibody. The results of Western blotting indicated that PAK5 can be highly expressed in the dental germ cells, suggesting that PAK5 may play an important role in biological function of dental germ cells.

[Determination of organic residues in macroporous adsorption resins by gas chromatography].

A method has been developed for the simultaneous determination of benzene, toluene, xylene, styrene, decane, diethylbenzene, undecane, divinylbenzene, dodecane and naphthalene residues in macroporous adsorption resins (MARs). The organic residues were extracted from resin samples by ultrasonic extraction with dichloromethane and analyzed by gas chromatography. The chromatographic conditions were as follows: a DB-624 capillary column (30 m x 0.32 mm i.d., 1.8 microm); carrier gas, nitrogen with a flow rate of 2.5 mL/min; column temperature, programmed from 40 degrees C to 200 degrees C at a rate of 14 degrees C /min and kept at 200 degrees C for 1 min; flame ionization detector temperature, 250 degrees C; injector temperature, 220 degrees C; splitless injection, 1 microL. All the 10 organic residues were separated well in 12 min. The recoveries for spiked standards (n = 3) were 73.8% - 107.9%. The relative standard deviations were 1.3% - 4.4%. The limits of detection were 0.007 - 0.03 mg/L. This method is sensitive, accurate and quick. Nine commercial MARs and their pretreated samples were assayed, and the results show that the contents of organic residues varied greatly in two kinds of samples. And the pretreated MARs can be used safely in the production of traditional Chinese medicines.