OBJECTIVETo make a mechanical analysis on three-dimensional finite element models of the mandibular first molar with the maximum distal occlusal (DO) structure defect after the root canal therapy and filling and crown restoration under static and impact loads and to provide a guideline for planning restoration for the clinic.

METHODSThe research adopted reverse engineering technology to build the model of three-dimensional finite element. The form of the intercuspal occlusion and cusp to cusp occlusion during the circulation of posterior teeth occlusion movement were simulated. Half-sine pulse/impact was chosen for the impact dynamic. The impact ratio was indicated to the stress change between impact loads and static loads.

RESULTSUnder the two kinds of loads, the maximum Mohr stress values of the metal crowns were shown in all models. The restoration effects between the two kinds of models were compared, the maximum Mohr stress value of the crown metal and dentin was not obviously difference. The maximum Mohr stress values of dentin were all obviously smaller than the stretch limit strength of dentin. The impact ratio closed to 1.

CONCLUSIONThe impact loads accorded with the oral actual situation more than the static loads, but the suitable analysis of the static loads could be accepted. The restoration of metal crown is necessary. The effects between the amalgam filling and full crown restoration and composite resin filling and full crown restoration is not difference obviously.

Composite Resins ; Crowns ; Dentin ; Finite Element Analysis ; Humans ; Molar

OBJECTIVETo establish a method for diagnosis of freshwater drowning by amplifying gyrB and 16S rRNA genes of Aeromonas hydrophila using PCR coupled with capillary electrophoresis (CE).

METHODSDNA samples were extracted from human, 18 planktons (including Candida albicans, Aeromonas hydrophila, and 16 species of algae), and 30 cases of tissue samples (including the lung, liver, and kidney, all examined with microwave digestion-vacuum filtration-automated scanning electron microscopy) from human cadavers, including 28 freshwater drowning victims and 2 with natural death. The DNA samples were amplified with the primer AH (for gyrB gene) and primer Ah (for 16S rRNA gene), and the products were analyzed with CE.

RESULTSPCR amplification followed by CE yielded negative results for DNA of human, Candida albicans and 16 species of algae, whereas a positive result was found for Aeromonas hydrophila DNA with PCR products of 195 bp (with primer AH) and 350 bp (with primer Ah). In the 28 drowning cases, the detection rates of Aeromonas hydrophila using primer AH were 96.4% in the lung tissue, 71.4% in the liver tissue, and 60.7% in the kidney, as compared with the rates of 75.0%, 42.9%, and 32.1% using primer Ah, respectively. The positive rates for Aeromonas hydrophila in the organs of the drowning victims were 82.1% and 53.6% with primer AH and primer Ah, respectively. The detection showed negative results in the 2 cases of natural deaths. The two primers produced significantly different detection rates of Aeromonas hydrophila (P<0.05).

CONCLUSIONPCR coupled with CE for detecting gyrB gene of Aeromonas hydrophila has a high sensitivity in assisting a diagnosis of freshwater drowning. Detection of both the gyrB gene and 16S rRNA gene of Aeromonas hydrophila can yield more convincing evidence of the diagnosis of freshwater drowning.

Objective To establish a Real-time quantitative PCR method (qPCR) for the detection of diatom UPA barcoding genes and evaluate its application in the drowning diagnosis. Methods The homologous sequences of diatoms UPA gene was obtained by Blast from GeneBank, based on which the universal primers for diatoms were designed. DNA were extracted from 2 common human symbiotic bacteria (Escherichia coli, Bifidobacterium longum), 3 species of planktonic bacteria, 15 species of planktonic algae, tissue samples (lung, liver and kidney) from human cadavers (28 drowning victims, 1 victims by non-drowning in the water, 3 victims deaths on land) in 32 cases. The specificity, sensitivity and repeatability of the designed primers were tested. The positive rates of diatoms detection in the drowning cases were calculated. The results of the real-time quantitative method were evaluated comparatively by Microwave Digestion-Vacuum Filtration-Automated Scanning Electron Microscopy (MD-VF-Auto SEM) and PCR-Capillary Electrophoresis (PCR-CE). Results The results showed that the primers UPA99 had strong specificity for the diatomaceae (Synedra radians, Navicula sp., Melosira varians, Cyclotella sp. and Nitzschia sp.) DNA. The melting curve of the amplified product was smooth; the peak was narrow; the melting temperature was (87±1)℃. The sensitivity of qPCR method was 1.56×10-5ng/μL with the detection range of 1.56×102ng/mL~1.56×10-5ng/μL, in contrast with the PCR-CE method (1.56×10-3ng/μL). This real-time PCR method showed high repeatability and stability with the coefficient of variation less than 2%. The detection rate of lung, liver and kidney was 89.3%, 71.4% and 64.3% respectively. Conclusion The established qPCR method, based on the universal primers designed for diatom UPA gene, has high specificity, high sensitivity and good repeatability. With a promising prospect for application, qPCR is suitable for drowning diagnosis.