This study was aimed to explore the features of clustered regularly interspaced short palindromic repeats (CRISPR) structures in Shigella by using bioinformatics. We used bioinformatics methods, including BLAST, alignment and RNA structure prediction, to analyze the CRISPR structures of Shigella genomes. The results showed that the CRISPRs existed in the four groups of Shigella, and the flanking sequences of upstream CRISPRs could be classified into the same group with those of the downstream. We also found some relatively conserved palindromic motifs in the leader sequences. Repeat sequences had the same group with corresponding flanking sequences, and could be classified into two different types by their RNA secondary structures, which contain "stem" and "ring". Some spacers were found to homologize with part sequences of plasmids or phages. The study indicated that there were correlations between repeat sequences and flanking sequences, and the repeats might act as a kind of recognition mechanism to mediate the interaction between foreign genetic elements and Cas proteins.
Base Sequence ; Clustered Regularly Interspaced Short Palindromic Repeats ; Computational Biology ; Genome, Bacterial ; Plasmids ; Shigella ; genetics

Objective : To learn the passive smoking exposure and hazard awareness among the residents aged 18 years and over in Zhengzhou,so as to provide evidence for tobacco control. Methods: By multi-stage stratified and clustered sampling method,the residents aged 18 years and over in Zhengzhou were selected. From June to October in 2018,a questionnaire for risk factors of non-communicable diseases,designed by Chinese Center for Disease Control and Prevention,was used to collect their passive smoking exposure and hazard awareness and then analyzed. Results : A total of 6 793 questionnaires were qualified in 6 809 questionnaires and the effective rate was 99.77%. Among 5 387 non-smokers,2 131 people were exposed to passive smoking,and the crude and standardized rate was 39.56% and 40.14%. The standardized rate of passive smoking exposure was higher in men than in women（42.44% vs. 38.67%,P<0.05）,in rural residents than in urban residents（43.90% vs. 36.62%,P<0.05）,and it decreased with the age increase（P<0.05）. The standardized rate of passive smoking exposure in family,indoor workplace and indoor public place was 30.99%,36.99% and 68.02%,respectively. The standardized awareness rate of “passive smoking exposure may lead to heart disease,lung disease in children and lung cancer in adults” was 56.63%,which was higher in urban rural residents than in rural residents（75.69% vs. 36.33%,P<0.05）. Conclusions The passive smoking exposure rate was high in Zhengzhou,especially in indoor public places. The awareness of the hazards of passive smoking exposure was scarce, especially in rural residents.

Objective To detect the distribution of clustered regularly interspaced short palindromic repeat(CRISPR)associated protein genes cas1 and cas2 in Shigella and to understand the characteristics of CRISPR with relationship between CRISPR and related characteristics on drug resistance. Methods CRISPR associated protein genes cas1 and cas2 in Shigella were detected by PCR,with its products sequenced and compared.Results The CRISPR-associated protein genes cas1 and cas2 were found in all the 196 Shigella isolates which were isolated at different times and locations in China. Consistencies showed through related sequencing appepared as follows:cas2,cas1 (a) and cas1(b)were 96.44%,97.61%and 96.97%,respectively. There were two mutations including 3177129 site(C→G)and 3177126 site(G→C)of cas1(b)gene in 2003135 strain which were not found in the corresponding sites of Z23 and 2008113. Results showed that in terms of both susceptibility and antibiotic-resistance,strain 2003135 was stronger than Z23 and 2008113. Conclusion CRISPR system widely existed in Shigella,with the level of drug resistance in cas1(b) gene mutant strains higher than in wild strains. Cas1(b)gene mutation might be one of the reasons causing the different levels of resistance.

Objective This study aims to analyze the prevalence, distribution, and risk factors of abnormal glucose metabolism in urban and rural adult hypertension patients in Zhengzhou. Methods The study was conducted on permanent residents aged 18 years and over (with at least 6 months of residence) in urban and rural areas of Zhengzhou. The survey subjects were chosen by the multi‐stage sampling method. Basic information regarding the height, weight, waist circumference, and blood pressure of 6 798 people who met the standards of admission and discharge were obtained through an on‐site questionnaire survey and physical examination. Blood samples were collected and blood glucose and lipid levels were detected. Results There were 1 985 patients with hypertension, with a prevalence rate of 29.20% and the standardized prevalence rate was 26.53%. Blood samples were collected from 1 936 hypertensive patients (55.8±13.6 years), of whom 903 were male (46.64%) and 1 033 female (53.36%). Among 1 050 patients (54.24%) with abnormal glucose metabolism, the impaired fasting glucose rate was 4.44%, the decreased glucose tolerance rate was 22.21%, and the diabetes rate was 27.56%. Blood samples were collected from 4 748 non‐hypertensive patients and 1 123 patients (23.65%) with abnormal glucose metabolism. The prevalence of abnormal glucose metabolism in hypertensive patients was significantly higher than that in non‐hypertensive patients, and the difference was statistically significant (χ2=586.264, P<0.001). The prevalence rate of abnormal glucose metabolism in hypertensive patients was higher in females, 58.57%, than in males, 49.28% (χ2=16.743, P<0.001). Urban patients showed a higher abnormal glucose metabolism, 57.68%, than rural patients did, 51.67% (χ2=6.984, P<0.001), and the prevalence of abnormal glucose metabolism in hypertensive patients showed a significant upward trend with an increase in age. Patients with central obesity, abnormal blood lipids, family diabetes history, non‐smokers, and non‐drinkers showed a higher prevalence of abnormal glucose metabolism, and the differences were statistically significant (P<0.05). The prevalence of abnormal glucose metabolism increased with age and body mass index, and decreased with increase in education level, with differences being statistically significant (P<0.001). Multivariate logistic regression analysis showed that age, obesity, centripetal obesity, dyslipidemia, and family history of diabetes were the main risk factors for abnormal glucose metabolism in patients with hypertension. Conclusions There is a relatively high prevalence of abnormal glucose metabolism in hypertension patients among the urban and rural residents of Zhengzhou. In the management of hypertension, the detection and intervention for reducing abnormal glucose metabolism should be strengthened and related risk factors must be prevented. The age for health management objectives among hypertension and diabetes patients should be further reduced.

OBJECTIVETo detect the distribution of clustered regularly interspaced short palindromic repeat (CRISPR) associated protein genes cas1 and cas2 in Shigella and to understand the characteristics of CRISPR with relationship between CRISPR and related characteristics on drug resistance.

METHODSCRISPR associated protein genes cas1 and cas2 in Shigella were detected by PCR, with its products sequenced and compared.

RESULTSThe CRISPR-associated protein genes cas1 and cas2 were found in all the 196 Shigella isolates which were isolated at different times and locations in China. Consistencies showed through related sequencing appeared as follows: cas2, cas1 (a) and cas1 (b) were 96.44%, 97.61% and 96.97%, respectively. There were two mutations including 3177129 site(C→G)and 3177126 site (G→C) of cas1 (b) gene in 2003135 strain which were not found in the corresponding sites of Z23 and 2008113.

RESULTSshowed that in terms of both susceptibility and antibiotic-resistance, strain 2003135 was stronger than Z23 and 2008113.

CONCLUSIONCRISPR system widely existed in Shigella, with the level of drug resistance in cas1 (b) gene mutant strains higher than in wild strains. Cas1 (b) gene mutation might be one of the reasons causing the different levels of resistance.

Bacterial Proteins ; genetics ; CRISPR-Associated Proteins ; genetics ; Clustered Regularly Interspaced Short Palindromic Repeats ; Drug Resistance, Bacterial ; genetics ; Mutation ; Shigella ; genetics

OBJECTIVETo detect the molecular characteristics of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) in Shigella and to analyze the distribution of CRISPR related to the time of isolation.

METHODSOf the 52 Shigella strains, 41 were isolated from Henan, 6 from Jiangxi and 5 isolated from Beijing. Both CRISPR locus of S1, S2, S3 and S4 in Shigella were detected by polymerase chain reaction (PCR). The PCR products were sequenced and compared.

RESULTSThe positive rates of CRISPR locus in Shigella were 33.69% (S1), 50.00% (S2), 82.69% (S3) and 73.08% (S4), respectively. Two subtypes were discovered in S1 and S3 locus. Three subtypes were discovered in S2 locus. Four different subtypes were discovered in S4 locus. The isolates from Henan strains were divided into two groups by the time of isolation. Distributions of S1 were different, before or after 2004, on Shigella. S1 could not be detected after 2004. There were no statistical differences of S2, S3 and S4 in two groups.

CONCLUSIONDifferent CRISPR subtypes or Shigella were discovered. A significant correlation was noticed between the CRISPR S1 related to the time of isolation but not between S2, S3 or S4 on the time of isolation.

Clustered Regularly Interspaced Short Palindromic Repeats ; Polymerase Chain Reaction ; Shigella ; genetics