Objective To evaluate the bactericidal effect of diode laser on Porphyromonas gingivalis (Pg),and to explore an optimized protocol for a safe dose of photodynamic therapy (PDT) to eliminate periodontal pathogens as well as the impact on the implant surfaces,so as to provide theoretical and experimental basis for PDT in periimplantitis therapy.Methods Artificial in vitro models were formatted by culturing Pg standard strain and ITI (International Team for Implantology) implants together in CDC broth.Then artificial in vitro models were treated by different doses of hematoporphyrin monomethyl ether (HMME) and different energy density of laser (EDL) for 60 s.The cultures were counted by colony form unit (CFU),and SPSS 17.0 statistical software was used for data statistical analysis to select the best EDL and HMME dose.Finally,ITI implants were observed by scanning electron microscope (SEM) to evaluate the impact of HMME-PDT on Pg of implant surfaces.Results When EDL was 12 J/cm2 and mass concentration of HMME was 25 μg/ml,SEM observations showed that PDT could effectively kill Pg ((13.00±5.00) CFU)without damaging the implant surfaces.Conclusions PDT therapy combining 630 nm diode laser with photosensitizer HMME have good bactericidal effect on Pg,and the EDL and HMME dose is as small as the clinical applicable safe dose.

AIDS Vaccines ; administration & dosage ; genetics ; immunology ; Animals ; HIV Infections ; immunology ; prevention & control ; virology ; HIV-1 ; genetics ; immunology ; Humans ; Immunity, Mucosal

In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.
Cell Line ; DNA Transposable Elements ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection

Objective To explore the psychological availability of the full-time candidates for master degree of nursing during the clinical work. Methods A phenomenological approach was adopted in this study. About 11 full-time candidates for master's degree of nursing took part in the semi-structured and in-depth interviews and the acquired data were analysed. Results About 4 themes promoting psychological availability were identified: strong research knowledge and skills, career development confidence, perception of attention from leaders and more satisfactory salary. The four themes hindering psychological availability were:clinical work tasks, lower level of expertise, lack of social support and lower perception of self benefit. Conclusions The full-time candidates for master's degree hold active perception and much expectation about their clinical work but meanwhile they feel passive and disappointed. Nursing administrators should develop distinguishing and definite clinical personnel training and continuing education programs for them, offer them an access to learning chances and emotional support in order to improve the psychological availability and stabilize the nursing team.

Aim To investigate Jianpi Qinghua Chinese herbal compound( JQCC) on the expressions of the rel-evant proteins of TLR4 and its downstream MyD88-de-pendent pathways, and on the inflammatory factor TNF-α in the animal model of chronic atrophic gastritis ( CAG) in rats, so as to discuss the molecular mecha-nism of JQCC in the treatment of CAG. Methods 53 Wistar rats were randomly divided into the blank con-trol group(n=8) and the CAG model group(n=45), and the animal model of CAG in rats was replicated by the “ammonia + sodium deoxycholic acid + ethanol”method. After the successful modeling was confirmed, the rest of the 40 CAG rats in the CAG model group were divided into the model group, the vitacoenzyme-tablet group, the low dose of JQCC group, the medium dose of JQCC group, the high dose of JQCC group ( each group n =8 ) . The experimental animals of all the groups were given intragastric administration of medication continuously for 30 days. Then the patho-logical histological changes were observed by HE stai-ning. The protein expressions of TLR4, MyD88, NF-КB and COX-2 were tested by Western-blot assay. And the serum TNF-α level was measured by ELISA. Results The protein expressions of TLR4 , MyD88 , NF-КB and COX-2 and the serum TNF-α level in the rats in the model group were increased evidently ( P<0. 01). Compared with the model group, the gastric mucosa lesions were improved in the low dose of JQCC group, the medium dose of JQCC group, the high dose of JQCC group, together with significant decreases of the protein expressions of TLR4 , MyD88 , NF-κB and COX-2 and the serum TNF-α level ( P <0. 05 or P <0. 01 ) . Conclusion JQCC could effectively improve the pathological and histological changes in the gastric mucosa in CAG rats, and the therapeutic mechanism might be related to the expressions of the relevant pro-teins of TLR4-MyD88-dependent pathways and the ex-pressions of anti-inflammatory cytokines.

Objective To study the setup accuracy and other factors during frameless stereotactic radiosurgery (SRS) of intracraial tumor using ExacTrac X-ray image guide system.Methods Totally 119 intracranial tumor patients from August 2014 to February 2016 underwent auto setup with infrared marker.Bilateral oblique cross field images were obtained with ExacTrac X-ray system,and went through comparison,registration and correction with the digitally reconstructed ones of the planning system.Then the translation accuracy errors at LAT,LNG and VRT directions and corresponding rotational accuracy errors were acquired,and the errors experienced extended analysis.Results The translation errors at LAT,LNG and VRT directions acquired with ExacTrac X-ray image guidance system and 6DOF couch were (0.16±0.13) mm,(0.17±0.14) mm and (0.15±0.11) mm respectively,the corresponding rotational errors were (0.21±0.15),(0.18±0.15),(0.18±0.14)° respectively,and the vector error was (0.32±0.16) mm.All of 3 translation and 3 rotational errors were in the SRS error range.Conclusion ExacTrac X-ray image guidance system combined with 6 degrees-of-freedom couch increases the treatment accuracy during frameless SRS,and thus is worthy promoting practically.

Objective To study the effects of self-management education on quality of life in children with asthma.Methods Seventy-five out-patients and in-patients with asthma were randomlydivided into 2 groups.The educational group(38 cases)received the basic knowledge education of asthma and self-management,while the control group received only the basic knowledge education of asthma.A inquiry was conducted in all patients using a Chinese PAQLQ(pediatric asthma quality of life questionnaire)before and 6 months after the treatment.The correlation and difference between 2 groups were analyzed.Results There were no significant difference before the therapy.After 6 months's treatment,there were uptrends on scorecard in some fields in such as symptom,activity,emotion and the total score in all patients(P

BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

OBJECTIVETo investigate the relationship among oxygen concentration, quorum sensing system, type secretion system, and biofilm production of Pseudomonas aeruginosa.

METHODSA total of 23 clinical strains of Pseudomonas aeruginosa were cultured at different levels of environmental oxygen for three days. Then biofilm mass and alginate were quantified. The expression levels of LasI and RhlI were detected by real time polymerase chain reaction (PCR). The secretion of exoenzyme S was examined by Western blot.

RESULTSBoth the biofilm mass (R=0.455, P=0.000) and alginate (R=0.367, P=0.000) were positively correlated with oxygen concentration. Real time PCR showed that the expression levels of LasI and RhlI were not significantly correlated with oxygen concentration (R=0.025, P=0.794; R=-0.044, P=0.653), the production of biofilm (R=0.001, P=0.990; R=0.011, P=0.909), or alginate(R=0.029, P=0.770; R=0.193, P=0.064). Western blot showed that the optimal oxygen concentration range for exoenzyme S secretion of Pseudomonas aeruginosa ranged 10% to 30%.

CONCLUSIONSHyperoxia can promote the production of biofilm and alginate by Pseudomonas aeruginosa. Las/Rhl system may not participate in biofilm production at the early stage due to the low bacteria amount. The increased production of biofilm may inhibit the expression of Type Secretion system and thus inhibit bacterial virulence.

Alginates ; metabolism ; Biofilms ; drug effects ; Oxygen ; metabolism ; Pseudomonas aeruginosa ; metabolism ; physiology ; Quorum Sensing ; drug effects ; physiology

Objective To study the relationship between ischemic stroke and gene polymorphism of an-giotensin-converting enzyme (ACE), apolipoprotein E (APOE) and methylene tetrahydrofolate reductase (MTHFR) among the population of Zhuang nationality in western Gui. Methods We directly sequenced ACE, APOE and MTHFR genes in 149 cases of ischemic stroke and 109 cases of normal people in western Gui. χ2 test was used to measure the relationship between gene polymorphism and ischemic stroke. Hardy-Weinberg genetic equilibrium test was used to evaluate the reliability of these data. Results In the ischemic stroke group, 62 cases, 22 cases and 65 cases carried II genetype, DD genetype and ID genetype in ACE. χ2 test showed no relationship between ACE gene polymorphism and ischemic stroke. In analysis of the polymorpism of APOE in the ischemic stroke and control group, no relationship between APOE gene polymorphism and ischemic stroke was found by χ2 test. MTFHR gene polymorphism was significantly related with ischemic stroke by χ2 test (P = 0.019). Conclusion Polymorphism of gene MTFHR but neither ACE nor APOE is significantly associated with ischemic stroke.