A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

Objective To investigate the effects of heart displacement on hemodynamics during off-pump coronary artery bypass grafting (OP-CABG) while the sites for anastomosis were being exposed. Methods Forty-seven patients of both sexes (36 male, 11 female) aged 50-82 years undergoing OP-CABG were enrolled in the study. Preoperative cardiac function was assessed : class Ⅱ in 22 patients; Ⅲ in 23 and Ⅳ in 2 according to NYHA classification.The mean ejection fraction was 0.55?0.14 before surgery.They received on average 3.2 grafts. Premedication consisted of intramuscular morphine 10 mg, midazolam 3-5 mg and scopolamine 0.3 mg.Before induction of anesthesia ECG and SpO2 were monitored and radial artery was cannulated for continuous direct BP monitoring. Anesthesia was induced with midazolam 0.1 nig?kg-1 , fentanyl 4?g?kg-1 and pancuronium 0.1 mg?g-1 iv.The patients were mechanically ventilated after tracheal intubation and PETCO2 was maintained at about 40 mm Hg. Anesthesia was maintained with isoflurane and 50%-60% N2O in O2 and intermittent intravenous boluses of fentanyl and pancuronium. Swan-Ganz catheter which can continuously monitor mixed venous blood O2 saturation (SvO2) was placed in pulmonary artery via right internal jugular vein. SvO2, cardiac output (CO), BP, pulmonary arterial pressure (PAP) and HR were continuously monitored. Right atrial pressure (RAP) and PAWP were measured intermittently. Cardiac index (CI),stroke index (SI),systemic vascular resistance index (SVRI),PVRI, left and right ventricular work index (LVWI,RVWI) and left and right ventricular stroke work index (LVSWI,RVSWI) were calculated. The hemodynamic parameters were recorded after induction of anesthesia before surgery (T1,baseline),before heart displacement (T2), while anastomosis to anterior descending branch was being made (T3), while anastonosis to right coronary artery or posterior descending branch (T4) and to left circumflex artery or diagonal branch (T5) was being made, after normal heart position was resumed (T6) and at the end of operation (T7). Results While anastomosis to the anterior descending branch was being made (T3) SI and LVSWI significantly decreased as compared with the baseline (P

Objective:To investigate the incidence of asthma and the characteristics of acute attacks in children aged 3-14 in different administrative areas of Wuxi, and to analyze the correlation of asthma with the outdoor environmental factors.Methods:A total of 10 175 children aged 3-14 were randomly selected from 5 different administrative regions in Wuxi City, Jiangsu Province for childhood asthma epidemiological survey from January to December in 2017.Results:The complete questionnaires were collected from 8 318 children with a response rate of 81.70%.There were 6 734 valid questionnaires.Asthma occurred to 259 children, and the prevalence rate was 3.84%.The prevalence of asthma in male and female children was 4.89% (163 cases) and 2.82% (96 cases), respectively.The ratio of male patients to female patients was 1.70∶1.00, and the difference was significant ( χ2=19.72, P<0.01). Children were most prone to suffer from asthma attacks at the turn of seasons (87/259 cases, 33.59%). The frequency of asthma was the lowest from June to August (9/259 cases, 3.47%). Among 244 cases attacked by acute asthma, the prevailing time period of acute attacks was irregular in 88 patients (36.07%), bedtime in 64 cases (26.23%). In different regions of the city, the lowest prevalence rate of asthma was found in Binhu District (50/1 830 cases, 2.73%), the highest prevalence rate of asthma was found in Xinwu District (71/1 502 cases, 4.72%), the difference between the highest and lowest prevalence rates of asthma was significant ( χ2=13.19, P<0.05). In terms of the air quality in different admi-nistrative regions of Wuxi, the nitrogen dioxide (NO 2) concentration, the carbon monoxide (CO) concentration, PM 10 and PM 2.5 in Binhu District were lower than other 4 regions.In different seasons in Wuxi city, the sulfur dioxide(SO 2) concentration, the NO 2 concentration, the CO concentration, PM 10 and PM 2.5 from June to August were significantly lower than those in other months.The prevalence of asthma in children in different administrative districts of Wuxi was weakly positively correlated with the SO 2 concentration, the NO 2 concentration and PM 10 ( r=0.10, P<0.01; r=0.22, P<0.01; r=0.06, P<0.01, respectively). The prevalence of asthma was weakly negatively associated with the ozone(O 3) concentration ( r=-0.06, P<0.01). Acute asthma attacks were weakly positively correlated with the SO 2 concentration ( r=0.22, P<0.01), the NO 2 concentration ( r=0.28, P<0.01), the CO concentration ( r=0.23, P<0.01), PM 10 ( r=0.18, P<0.01) and PM 2.5 ( r=0.18, P<0.01), and weakly negatively correlated with the O 3 concentration ( r=-0.40, P<0.01). Conclusions:The prevalence of asthma in Wuxi is higher in boys than in girls, and the frequency of asthma attacks is related to the season.The SO 2, NO 2, CO, PM 10, PM 2.5 and O 3 concentration affect the prevalence and acute attacks of asthma.

OBJECTIVETo evaluate the effect of chewing sugar-free gum after sucrose challenge on dental plaque pH in situ.

METHODS16 healthy volunteers aged 23 - 32 years were screened as subjects. The pH of 48-hour dental plaque was measured using a Beetrode pH microelectrode when subjects chewed Extra sugar-free gum after sucrose challenge.

RESULTSDental plaque pH maintained at resting plaque pH when immediately chewed sugar-free gum after sucrose challenge. Chewing sugar-free gum at 5 min after sucrose challenge, dental plaque pH was raised from 5.59 (measured at 5 min after sucrose challenge) to 6.98 (measured at 10 min after sucrose challenge).

CONCLUSIONSChewing sugar-free gum after sucrose challenge can neutralize organic acid produced by bacteria in dental plaque and rapidly rise plaque pH.

Adult ; Chewing Gum ; Dental Plaque ; metabolism ; prevention & control ; Female ; Humans ; Hydrogen-Ion Concentration ; Male ; Sucrose

OBJECTIVETo detect Porphyromonas gingivalis (P. gingivalis) in buccal epithelial cells and subgingival plaque from periodontally healthy subjects and patients with chronic periodontitis.

METHODS40 subjects were included in the healthy group and 39 subjects were included in the diseased group in this study. Cells and subgingival plaque samples were collected. The extracted DNA was amplified with universal primers and P. gingivalis species-specific primer.

RESULTSP. gingivalis was detected in 37.5% of subgingival plaque samples and 32.5% of buccal mucosa samples in the healthy subjects, but 69.23% of subgingival plaque samples and 46.15% of buccal mucosa samples in the periodontitis group. Highly statistically significant differences were observed between healthy and periodontitis groups in the detections of P. gingivalis of subgingival plaque samples.

CONCLUSIONP. gingivalis may be one of oral flora because it can be detected in the healthy population and not lead to destruction of supporting structures of the teeth.

Adult ; Bacteria ; Chronic Periodontitis ; Dental Plaque ; Epithelial Cells ; Humans ; Periodontitis ; Porphyromonas gingivalis

OBJECTIVETo study the molecular epidemiology of Vibrio parahaemolyticus isolates from clinical and environmental samples collected in Hangzhou area during 2000 and 2002.

METHODSV. parahaemolyticus isolates from food-poisoning, sporadic diarrhea patients and seafood in market in Hangzhou during 2000 and 2002 were serotyped. From clinical isolates of serotype O3:K6 and environmental isolates of serotype O3:KUT, virulence genes, tdh and trh, were examined by PCR. Molecular typings [including ribotyping, random amplified polymorphic DNA analysis (RAPD), enterobacterial repetitive intergenic consensus PCR (ERIC-PCR)], were also performed.

RESULTSAmong 13 food-poisoning outbreaks caused by V. parahaemolyticus, O3:K6 strains with tdh positive and trh negative were detected in 11 episodes (84.6%), and O4:K8 strains with tdh positive and trh negative were responsible for other 2 outbreaks (15.4%). In 34 V. parahaemolyticus isolates from sporadic diarrhea patients, O3:K6, O4:K8, and O1:KUT strains with tdh positive and trh negative were detected with proportions of 26.5% (9/34), 17.6% (6/34), and 38.2% (13/34), respectively, and other 6 strains were not able to be serotyped. Of 64 isolates from seafood in which both tdh and trh were negative except trh was positive in one O1:KUT strain, 37 belonged to 7 serotypes except O3:K6 or O4:K8, and 9 were not able to be serotyped. The fingprintings of ribotyping, ERIC-PCR, and RAPD showed that almost all of O3:K6 isolates from food-poisoning and sporadic diarrhea patients were genetically close to each other and there were obviously genetical difference between O3:K6 isolates from clinic and O3:KUT isolates from environment.

CONCLUSIONThere was a distinct serotype distribution between V. parahaemolyticus isolates from clinic patients and seafood. One group of close related V. parahaemolyticus O3:K6 strains with tdh positive and trh negative seemed to be prevailing in Hangzhou in those years.

China ; epidemiology ; DNA, Bacterial ; analysis ; Disease Outbreaks ; Food Contamination ; Foodborne Diseases ; Humans ; Molecular Epidemiology ; Polymerase Chain Reaction ; Seafood ; Vibrio parahaemolyticus ; genetics ; isolation & purification

OBJECTIVETo study the effects of Porphyromonas gingivalis (Pg) on the proliferation and apoptosis of vascular endothelial cell and to identify the role of Pg in the pathogenesis of atherosclerosis.

METHODSA cell culture model of vascular endothelial cell treated by Pg was used in vitro. The cells' ability of proliferation was measured by methyl thiazolyl tetrazolium assay, and cell cycle and the percentage of apoptosis with or without Pg invasion were examined by flow cytometry.

RESULTSInvasion of Pg could inhibit the proliferation of vascular endothelial cells at 72 h (F = 786.68, P < 0.01), and induce apoptosis at 24 h (F = 1074.56, P < 0.01). In addition, the cell cycle was arrested at G1 phase by the invasion of Pg W83 at 24 h (F = 43.23, P < 0.01) and ATCC 33277 at 48 h (F = 66.72, P < 0.01).

CONCLUSIONSPg may aggregate the inflammation of the vascular endothelial cells through induction of apoptosis, which could be one of the pathologic mechanisms in atherosclerosis.

Apoptosis ; Bacteroidaceae Infections ; physiopathology ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Porphyromonas gingivalis

OBJECTIVETo investigate the distribution of rag genotypes of Porphyromonas gingivalis (P. gingivalis) in chronic periodontitis patients.

METHODSSubgingival plaque samples were collected from 50 chronic periodontitis patients. The occurrence of P. gingivalis was determined by polymerase chain reaction (PCR) using 16S rDNA-specific primers. Distribution of rag genotypes was assessed in P. gingivalis positive samples by PCR.

RESULTSThe occurrence of P. gingivalis was 70.7%, and the distribution of four rag genotypes among P. gingivalis positive samples was as follows: rag-1 60.4%; rag-2 23.6%; rag-3 44.3%; rag-4 15.1%, respectively.

CONCLUSIONP.gingivalis with various rag genotypes was present in subgingival plaque samples from chronic periodontitis patients, and P. gingivalis with rag-1 and rag-3 were more predominant in chronic periodontitis patients, which may be associated with the development of periodontitis.

Adult ; Chronic Periodontitis ; DNA-Binding Proteins ; Dental Plaque ; Female ; Genotype ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis

To explore the growth and development and analyze the quality of the parthenocarpy fruit induced by exogenous hormones of Siraitia grosvenorii. the horizontal and vertical diameter, volume of the fruit were respectively measured by morphological and the content of endogenous hormones were determined by ELISA. The size and seed and content of mogrosides of mature fruit were determined. The results showed that the fruit of parthenocarpy was seedless and its growth and development is similar to the diploid fruit by hand pollination and triploid fruit by hand pollination or hormones. But the absolute value of horizontal and vertical diameter, volume of parthenocarpy fruit was less than those of fruit by hand pollination, while triploid was opposite. The content of IAA, ABA and ratio of ABA/GA was obviously wavy. At 0-30 d the content of IAA and ABA of parthenocarpy fruit first reduced then increased, content of IAA and GA parthenocarpy fruit was higher than that of fruit by hand pollination. Mogrosides of parthenocarpy fruit was close to pollination fruit. Hormones can induce S. grosvenorii parthenocarpy to get seedless fruit and the fruit shape and size and quality is close to normal diploid fruit by hand pollination and better than triploid fruit by hormone or hand pollination.
Cucurbitaceae ; chemistry ; drug effects ; genetics ; growth & development ; Diploidy ; Fruit ; chemistry ; genetics ; growth & development ; Plant Growth Regulators ; pharmacology

Dysentery, Bacillary ; epidemiology ; Humans ; Molecular Epidemiology ; Shigella ; genetics