Objective To study the genetic basis for biovar conversion of Y. pestis. Methods In silico comparative genomic analysis was conducted and some critical genetic variations of Yersinia pestis were comparatively analyzed by means of PCR and DNA sequencing. Results A 93bp in-frame deletion in glpD gene results in the glycerol negative characteristic of Orientalis strains. A point mutation in the napA gene may cause the negative characteristic of nitrate reduction in Mediaevalis and Microus strains. A 122-bp frameshift deletion in the araC gene may lead to the arabinose negative phenotype of Microus strains. Conclusion In this study, Microtus strains with their unique pathogenic, biochemical and molecular features, were proposed as a novel biovar Microtus. In the light of its differential ability to ferment glycerol and arabinose and to reduce nitrate, Y. pestis can be classified into four biovars-Antiqua(glycerol positive, arabinose positive and nitrate positive), Mediaevalis(glycerol positive, arabinose positive and nitrate negative), Orientalis(glycerol negative, arabinose positive and nitrate positive), and Microtus(glycerol positive, arabinose negative and nitrate negative).

Objective To better understand the pathogenicity and evolution of Yersinia pestis, we carried out the whole genome sequencing of human-avirulent Yersinia pestis strain 91001, which was isolated from a species of rodent-Microtus brandti. Methods We utilized “whole genome shotgun” approach to get the genome sequence of 91001. Based on the finished and annotated genome sequence of 91001, as well as the previously published genome sequences of CO92 and KIM, we performed detailed comparative genomics analysis on their chromosomes and plasmids. Results The genome of 91001 consisted of one chromosome and four plasmids (pPCP1, pCD1, pMT1 and pCRY). The pPCP1 plasmid of 9 609bp was almost identical with its counterparts from reference strains, which possessed 10 CDS. Plasmid pCD1 was found to be a plasmid of the type III secretory apparatus, and its length was 70 159bp. Although its CDS are quite similar to those of the reference plasmids, there were obvious rearrangements which produced certain differences in structure among them. Another plasmid was pMT1, a 106 642bp plasmid, which showed slightly different architecture compared with the reference ones. There was no mutation in virulent-related genes of pMT1 and pMT1 of 91001, which seemed to have retained more fragments of an ancestor plasmid. pCRY was a novel plasmid discovered in this work. It was 21 742bp long and harbored a group of gene encoding type IV secretory system. pCRY seemed to be able to replicate. The length of chromosome of 91001 was 4 595 065bp, and among its 4 037 predicted CDS (coding sequences), 141 were possibly pseudogenes. There were many IS in the chromosome. Due to the rearrangments mediated by IS, the structure of 91001 chromosome showed significant differences compared with CO92 and KIM. According to the results of comparative genomics analysis, we deduced the genetic mechanisms of nitrate reduction, glycerol fermentation, arabinose and milibiose utilization in 91001. Conclusion According to the analysis of plasmids structure, pseudogenes distribution, nitrate reduction negative mechanism, gene comparison and chromosome architecture, we conclude that 91001 and other strains isolated from Microtus brandti and Microtus fuscus evolved from ancestor Y. pestis and then developed into a different lineage. The deletion of large genome fragments from 91001 chromosome and pseuogenes might contribute to its unqiue pathogenicity and host-specificity.

Objective To establish reliable proteomics analysis models for Yersinia pestis and obtain the basic proteomics data of this pathogen. Methods The human-avirulent Y. pestis strain 91001 was cultured as an experimental strain, and the proteins of which were extracted and divided into three parts fractionally according to their solubility. Then three different methods (Shotgun-LC-MS-MS, 1D-LC-MS-MS and 2D-MS) were used to analyze the extracted proteins. The obtained data were compared with the theoretical protein database of strain 91001 so as to identify the expressed protein of Y.pestis strain 91001 in this study. Results 971 proteins were identified by shotgun-LC-MS-MS method, accounting for 23.4% of the predicted proteins of strain 91001 (971/4 143). 915 proteins were identified by 1D-LC-MS-MS method, accounting for 22.1% of the predicted proteins of strain 91001 (915/4 143). However, with 2D-MS only 5.62% of the predicted proteins (233/4 143) were identified. Altogether 1 193 proteins were identified when the results of the 3 methods added together, accounting for 28.7% of all the predicted CDS in 91001. Conclusion The kind and quantity of proteins identified by various proteomics methods differ from each other dramatically, therefore it is necessary to utilize multiple methods to get more reliable protein profiles of Y. pestis.

Objective To study genetic variations of pgm locus and its flanking regions in Yersinia pestis isolated from Chinese natural focus so as to understand differences in virulence between different strains and to improve the prevention of plague. Methods We analysed the sequence variations of pgm locus and its flanking regions by using PCR-sequencing and allele-specific PCR among 260 strains of Y. pestis isolated from different foci and 7 strains of Yersinia pseudotuberculosis. Results For YP1666, only the strains of Xilin Gol grassland type and Microtus fuscus type had intact transmembrane helix, and the T-deletion at base 1406 was unique in strains of Orientalis. Same as Y. pseudotuberculosis, there was no IS100-insertion at the 3′ end of pgm locus of strains from Eastern section of North Tianshan Mountain type, Western section of North Tianshan Mountain type A and B, Xilin Gol grassland type and M. fuscus type. Only the strains of Xilin Gol grassland type and M. fuscus type had IS285-insertion in pigmentation segment and IS100-insertion at its downstream flanking region. pgm locus was deleted entirely from 36 strains, most of which came from Ordos plateau type, Song-liao plain type B , Kunlun Mountain type A and B. Conclusion Strains of Eastern section of North Tianshan Mountain type, Western section of North Tianshan Mountain type A and B, Xilin Gol grassland type and M. fuscus type are the oldest lineage of Chinese isolates. The pgm locus of the strains of these three types is very stable because there is no IS100-insertion at its 3′ terminal. We suggest that the strains of Xilin Gol grassland type and M. fuscus type should be grouped into a fourth biovar: Microtus. pgm locus is highly conserved among strains of different ecotypes, and its variations are well correlated with biovar, focus and ecotype, which indicates that pgm locus has played a role in strains′ adaptation to their environment.

Objective To evaluate the relationship of the plasma levels of osteopontin(OPN),lipoprotein (a) [Lp (a)] and apolipoproteinA/apolipoproteinB (ApoB/ApoA1) ratio with atherosclerosis.Methods 120 elderly patients with atherosclerosis (observation group) and another 120 healthy elderly subjects (control group) from Health Check Department in Tianjin Third Center Hospital were enrolled from Mar.2014 to Nov.2015.The levels of plasma OPN,Lp(a) and ApoB / ApoA1 ratio were measured and their relationship with atherosclerosis was analyzed.Results In observation group versus control group,the levels of plasma OPN[(33.2±10.1) μg/L vs.(30.6± 9.1) μg/L],ApoB/ApoA1 ratio[(0.8±0.7) vs.(0.6±0.1)]and Lp(a)[(357.6±66.5) mg/L vs.(120.5±35.7) mg/L] were increased with statistically significant differences (t =8.91,8.43,6.01;P=0.021,0.035,0.041).The concentrations of plasma OPN,ApoB/ApoA1 ratio and Lp (a) were increased along with aggravation of artery lesion,with statistically significant differences (F =8.50,7.38,6.41;P=0.031,0.039,0.037).Conclusions The plasma levels of OPN,Lp(a) and ApoB/ ApoA1 ratio are increased in patients with atherosclerosis and increased along with the lesion aggravation.

Objective To study the roles of soluble intercellular adhesion molecule 1(sICAM 1) in the pathogenesis of pregnancy induced hypertension (PIH). Methods Serum levels of sICAM 1 were measured by enzyme linked immunosorbent assay(ELISA) in 65 women with PIH (PIH group : 15 mild PIH, 24 moderate PIH and 26 severe PIH) and 25 normal pregnant women (control group). The serum levels of IL 1? and TNF ? were measured by chemiluminescent ELISA at the same time. Results Serum concentrations of sICAM 1 were significantly higher ( P

Objective: To understand the status of health literacy and its associated factors among primary school students in Shandong Province. Methods: By using stratified cluster sampling method, 9 063 children in grade 1 to 3 from 5 cities in Shandong Province were investigated with self-designed questionnaire. Chi square and multivariate logistic regression were used to analyze associated factors for health literacy. Results: Rate of proficient health literacy among grade 1-3 primary school students was 62.8%(5 689/9 063), with 75.6%(6 850/9 063), 56.3%(5 106/9 063) and 80.1%(7 261/9 063) in dimension of safety emergency, health care and disease prevention and control. Proficient health literacy was 47.7%(4 327/9 063), 88.3%(8 003/9 063) and 73.6%(6 672/9 063) in domain of health knowledge and concept, health behavior and health skills. Univariate analysis showed that the health literacy varied by gender, only-child in the family, grade, living area, cities, self-perceived academic performance, father educational attainment, mother educational attainment, father occupation and mother occupation(χ2=39.66, 161.09, 193.05, 89.23, 1 107.75, 196.35, 99.43, 74.39, 64.86, 16.70, P<0.01). Multivariate analysis found that gender, only-child in the family, grade, living area, city, self-perceived academic performance, father’ s occupation and educational attainment were positively associated with health literacy among elementary students. Conclusion High level of health literacy has been found among primary school students in Shandong Province, but varied in area and different demographic characteristics, which needs further specific intervention.

Animals ; Biocatalysis ; Cattle ; Enteropeptidase ; chemistry ; metabolism ; Humans ; Models, Molecular ; Oryzias ; metabolism ; Structure-Activity Relationship ; Substrate Specificity

Objective:To explore any effects of combining virtual reality training with aromatherapy in caring for Alzheimer′s disease patients in a nursing home.Methods:Fifty nursing home residents with Alzheimer′s were divided at random into an observation group and a control group, each of 25. Both groups received routine rehabilitation, while the observation group was additionally given 45 minutes of virtual reality training combined with aromatherapy, 3 times a week for 6 months. Both groups′ cognition was then evaluated using the MMSE and an Alzheimer′s cognition assessment scale (ADAS-cog). Psycho-behavioral symptoms were quantified using the Alzheimer′s disease pathological behavior scale (BEHAVE-AD). Motor functioning was quantified using the timed up and go test (TUGT), the 30-second sit-to-stand test (30sCST), the 30-second arm curl test (30sACT) and the sit-and-reach test (CSRT). Ability in the activities of daily living (ADL) and life quality were quantified using the activity of daily living scale and of the quality of life scale for Alzheimer′s disease (QOL-AD) before and after the intervention.Results:After the intervention the average MMSE, ADAS-cog, BEHAVE-AD and ADL scores of both groups had improved significantly, with the average improvement in the observation group significantly greater than that in the control group. The TUGT, 30sCST, 30sACT and CSRT results of both groups were also significantly better, with those of the observation group again significantly superior, on average, to the control group′s results. The average QOL-AD score in the observation was significantly improved after the intervention, and was then significantly better than the control group′s average.Conclusions:Virtual reality training combined with aromatherapy can significantly improve the cognition, psycho-behavioral symptoms, activity in daily living, motor functioning and life quality of Alzheimer′s patients in a nursing home. It is worthy of promotion and application in nursing homes.

Objective:To observe any effect of virtual reality (VR) training on the cognitive functioning and functional fitness of nursing home residents with subjective cognitive decline (SCD).Methods:Fifty-six of such residents were randomly divided into an observation group and a control group, each of 28. Both groups received health education and routine care, but the observation group was additionally provided with 45 minutes of VR training three times a week for 6 months. The training included Baduanjin, magic, flying bird, supermarket shopping, gravity ball and gym episodes. Both groups′ cognition was evaluated using the subjective cognitive decline questionnaire (SCD-Q), the Montreal cognitive assessment (MoCA), the Rivermead Behavioural Memory Test (second edition) (RBMT-Ⅱ), a digit symbol substitution test (DSST), an animal fluency test (AFT) and trail-making test A-B (TMT A-B). Functional fitness was quantified using the 8-foot up-and-go test (8UGT), a 30-second arm curl test (30sACT), a 30-second chair stand test (30sCST), a back scratching test (BST), the sit-and-reach test (CSRT) and a 2-minute step test (2MST) before and after the 6-month intervention.Results:After the intervention, the average SCD-Q, MoCA, RBMT-Ⅱ, DSST, TMT-A, and TMT-B scores of the observation group were significantly better than before the intervention, and significantly better than the control group′s averages. And except for the back scratching their functional results were also significantly better, on average, than those of the control group.Conclusions:VR training can effectively improve the cognition and functional fitness of nur-sing home residents with SCD. Such training is worthy of promotion and wider application in nursing homes.