A Core Genome Multilocus Sequence Typing Scheme for Proteus mirabilis / 生物医学与环境科学（英文）

OBJECTIVE@#A core genome multilocus sequence typing (cgMLST) scheme to genotype and identify potential risk clonal groups (CGs) in Proteus mirabilis.@*METHODS@#In this work, we propose a publicly available cgMLST scheme for P. mirabilis using chewBBACA. In total 72 complete P. mirabilis genomes, representing the diversity of this species, were used to set up a cgMLST scheme targeting 1,842 genes, 635 unfinished (contig, chromosome, and scaffold) genomes were used for its validation.@*RESULTS@#We identified a total of 205 CGs from 695 P. mirabilis strains with regional distribution characteristics. Of these, 159 unique CGs were distributed in 16 countries. CG20 and CG3 carried large numbers of shared and unique antibiotic resistance genes. Nine virulence genes ( papC, papD, papE, papF, papG, papH, papI, papJ, and papK) related to the P fimbrial operon that cause severe urinary tract infections were only found in CG20. These CGs require attention due to potential risks.@*CONCLUSION@#This research innovatively performs high-resolution molecular typing of P. mirabilis using whole-genome sequencing technology combined with a bioinformatics pipeline (chewBBACA). We found that the CGs of P. mirabilis showed regional distribution differences. We expect that our research will contribute to the establishment of cgMLST for P. mirabilis.

Improving Effect of Modified Buyang Huanwutang Combined with Electroacupuncture on Traumatic Spinal Cord Injury due to Qi Deficiency and Blood Stasis / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Buyang Huanwutang combined with electroacupuncture （EA） in the treatment of traumatic spinal cord injury （TSCI） due to Qi deficiency and blood stasis. Method:Eighty-seven TSCI patients who met the inclusion requirements were randomly divided into an observation group （<italic>n</italic>=44） and a control group （<italic>n</italic>=43）. On the basis of comprehensive western medical treatments， patients in the control group were further provided with Wuwei Tongshuan oral liquid，10 mL per time，three times per day， while those in the observation group received modified Buyang Huanwutang，one bag per day，for 12 consecutive weeks. Besides， EA was performed in both groups in the same way， once per day， six times per week， for six weeks in total. The American Spinal Injury Association （ASIA） motor score， modified Barthel index （MBI），visual analog scale （VAS） pain score，Berg balance scale （BBS） score，modified Ashworth scale （MAS） score， spinal cord independence measure-Ⅲ（SCIM-Ⅲ） score， lower limb range of motion （ROM）， and Qi deficiency and blood stasis syndrome score before and after treatment were evaluated， followed by the recording of the occurrence of complications during treatment. The brain-derived nerve growth factor （BDNF）， nerve growth factor （NGF）， vascular endothelial growth factor （VEGF）， neurotrophic factor-3 （NT-3）， malondialdehyde （MDA） and superoxide dismutase （SOD） levels before and after treatment were determined. Result:The motor， light touch， needling sensation， MBI， and BBS scores of the observation group were higher than those of the control group （<italic>P</italic><0.01）， while the AS and MAS scores were lower（<italic>P</italic><0.01）. The angles of adductor and straight leg raising in the observation group were greater than those of the control group （<italic>P</italic><0.01），but the Qi deficiency and blood stasis syndrome score was lower（<italic>P</italic><0.01）. Both the scores of self-care， respiration， and sphincter management in SCIM-Ⅲ and the total score in the observation group were elevated as compared with those of the control group （<italic>P</italic><0.01）. The cumulative incidence of complications in the observation group was 34.09%，significantly lower than 55.81% in the control group （<italic>χ</italic>²=4.149，<italic>P</italic><0.05）. Compared with the control group， the observation group exhibited remarkably increased BDNF， NGF， VEGF， NT-3， and SOD （<italic>P</italic><0.01） and decreased MDA （<italic>P</italic><0.01）. Conclusion:Modified Buyang Huanwutang combined with EA is effective in alleviating spinal cord injury， promoting neural functional recovery， improving independence in activities of daily living， reducing the incidence of complications of patients with TSCI， which may be related to the amelioration of ischemia and hypoxia， inhibition of lipid peroxidation， and acceleration of nerve cell repair and regeneration.

Epidemiological Characteristics of Notifiable Infectious Diseases among Foreign Cases in China, 2004-2017 / 生物医学与环境科学（英文）

Objective@#We aimed to assess the features of notifiable infectious diseases found commonly in foreign nationals in China between 2004 and 2017 to improve public health policy and responses for infectious diseases.@*Methods@#We performed a descriptive study of notifiable infectious diseases among foreigners reported from 2004 to 2017 in China using data from the Chinese National Notifiable Infectious Disease Reporting System (NNIDRIS). Demographic, temporal-spatial distribution were described and analyzed.@*Results@#A total of 67,939 cases of 33 different infectious diseases were reported among foreigners. These diseases were seen in 31 provinces of China and originated from 146 countries of the world. The infectious diseases with the highest incidence number were human immunodeficiency virus (HIV) of 18,713 cases, hepatitis B (6,461 cases), hand, foot, and mouth disease (6,327 cases). Yunnan province had the highest number of notifiable infectious diseases in foreigners. There were different trends of the major infectious diseases among foreign cases seen in China and varied among provinces.@*Conclusions@#This is the first description of the epidemiological characteristic of notifiable infectious diseases among foreigners in China from 2004 to 2017. These data can be used to better inform policymakers about national health priorities for future research and control strategies.

Correlations of serum CEA and CA19-9 levels with clinical staging of colorectal cancer / 预防医学

Objective To evaluate the value of serum CEA and CA19-9 concentration for clinical staging of colorectal cancer. Methods A total of 350 patients who underwent the surgical treatments for colorectal cancer between February 2015 to January 2017 were enrolled. The serum CEA and CA19-9 were detected by chemoluminescence method. Results The positive rate of CEA of patients in stageⅠto Ⅳ was 25.00%, 36.69%, 50.78% and 66.67%, respectively. The positive rate of CA19-9 of patients in stageⅠto Ⅳwas 2.94%, 10.07%, 17.97% and 53.33%, respectively. The positive rates of CEA and CA19-9 were gradually increased with the stage developing (P<0.05). Results from multivariate logistic regression analysis showed that the positive levels of CEA and CA19-9 were risk factors in the TNM staging of colorectal cancer. The ORs and 95%CI were 1.790 (1.163-2.755)and 3.476(1.790-6.749), respectively. Conclusion The positive serum concentrations of CEA and CA19-9 showed significant associations with TNM staging. Preoperative serum concentrations of CEA and CA 19-9 could be auxiliary diagnostic indicators to assess the condition of colorectal cancer.

Pulsed-field gel electrophoresis typing on non-O157 Shiga toxin-producing Escherichia coli isolates / 中华流行病学杂志

<p><b>OBJECTIVE</b>To establish a database and to understand the molecular epidemiological features of non-O157 Shiga toxin-producing Escherichia coli (STEC) isolates from different animal reservoirs and patients.</p><p><b>METHODS</b>Pulsed-field gel electrophoresis (PFGE) was performed according to the PulseNet protocol with minor modifications. A dendrogram was constructed using the BioNumerics.</p><p><b>RESULTS</b>Under the PulseNet protocol, 62 PFGE patterns were obtained from 76 non-O157 STEC isolates and then divided into A to M groups. Isolates from different sources were widely distributed in different groups, but were predominant seen in certain groups.</p><p><b>CONCLUSION</b>The non-O157 STEC isolates in China were highly polymorphic. PulseNet protocol seemed to be suitable for the typing of Chinese non-O157 STEC isolates.</p>

An etiological survey on a foodborne disease epidemic outbreak caused by Salmonella enteritidis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To conduct an etiological molecular epidemiological survey and laboratory test on a foodborne disease epidemic outbreak to make clear of the cause and implement effective prevention and control on it.</p><p><b>METHODS</b>On May 12th 2012, 135 kindergarten children were sent to Xuzhou City People's Hospital and Children's Hospital with gastrointestinal infection disease. A total of 34 anus swab samples and 4 vomit samples were collected from the patients. Real-time PCR rapid detection, strains separation and cultivation, phage lysis experiments, ATB automated identification system were used to make etiological detection and identification. The genomic DNA of salmonella enteritidis were typed with the pulsed-field gel electrophoresis (PFGE), cluster analysis were carried out together with the patterns of local Salmonella infections.</p><p><b>RESULTS</b>Children in 20 classes were suffered from the gastrointestinal infection among the 21 classes. There were no significant aggregation of class distribution. Among the 135 patients, 76 were boys (56.3%) and 59 were girls (43.7%). The main symptoms were fever (above 38°C), diarrhea and bellyache. Through real-time PCR detection and strains separation, 19 salmonella enteritidis were isolated from 34 anus swab samples of suspected cases and the detection rate was 56%. There were no strains detected from vomit samples. All of the 19 salmonella enteritidis showed the same serological subtype, biochemical reaction, drug sensitivity and phage lysis pattern. The salmonella enteritidis had the identical PFGE pattern (100% similarity), and were different from the pattern of local sporadic infection cases.</p><p><b>CONCLUSION</b>It was confirmed that this was an epidemic outbreak of foodborne disease caused by homologous salmonella enteritidis by epidemiological survey, clinical information, lab etiological test and molecular typing.</p>

Development of a singleplex PCR assay targeting O-antigen modification genes for molecular serotyping of Shigella flexneri / 中华流行病学杂志

Objective To develop a singleplex PCR assay targeting O-antigen modification genes for molecular serotyping of Shigella (S.)flexneri.Methods Eight pairs of primer for O-antigen synthesis and modification genes of S.flexneriwere designed and used for developing an O-antigen modification gene-specific singleplex PCR assay to serotype 14 most common S.flexneri serotypes (1 a,1 b,1 c,2a,2b,3a,3b,4a,4b,5a,Y,X,Xv and F6).Bacterial pathogens which causing diarrheal disease were used for specificity detection.106 S.flexneri clinical isolates were serotyped by this method and compared with the slide agglutination method.Results An O-antigen modification,gene-specific singleplex PCR was developed.When six singleplex PCR reactions were performed,14 of the 15 recognized S.flexneri serotypes were identified,except for serotype Xv.The detection threshold ranged from 10 pg to 1 ng DNA in a 20 μ l reaction system.A high concordance between the singleplex PCR assay and slide agglutination were observed when 106 S.flexneri strains of various serotypes were analyzed with an exception that 1 serotype Y strain showed that it was carrying the additional defective gtr Ⅱ genes.Conclusion This method showed advantages over the traditional slide agglutination methods,and was promising when under application in the following situations as clinical diagnosis.

Study on the integration site and arrangement of SfII and SfX prophages in Shigella flexneri serotype 2b strains / 中华流行病学杂志

Objective To study the integration site and arrangement of SfII and SfX prophages in Shigella flexneri serotype 2b strains. Methods A series of primers were designed based on potential integration site of SfII and SfX prophages in Shigella flexneri serotype 2b strains, and PCR were performed for 50 serotype 2b strains to amplify special genes located in host and prophages. PCR products were sequenced to identify integration sites and arrangement of SfII and SfX. Results In all the serotype 2b strains, prophage SfII and SfX were adjacent to each other, and integrated into the thrW tRNA gene of the host, which were located between genes proA and yaiC of host. Prophage SfX was located immediately upstream of prophage SfII in all the detected 50 serotype 2b strains exception for strain 51251. Conclusion This was the first report on the integration site and arrangement of serotype-converting prophages SfII and SfX in Shigella flexneri 2b strains.

The POSSUM scoring system for risks assessment in pancreatoduodenectomy / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the value of the POSSUM scoring system in predicting postoperative morbidity and mortality of pancreatoduodenectomy (PD).</p><p><b>METHODS</b>Two hundreds and sixty-five consecutive PDs were performed between January 2005 and December 2007. POSSUM scores which relied on 12 physiologic and 6 operative variables were prospectively calculated for each case. Expected morbidity and mortality were estimated based on POSSUM scores and were compared with observed morbidity, which were diagnosed according to the Clavien complication scheme and domestic reference criteria respectively, and mortality.</p><p><b>RESULTS</b>Physiologic scores of 265 cases ranged from 12 to 24,the mean was 15. Operative scores ranged from 14 to 24, the mean was 17. The overall POSSUM scores ranged from 0.24 to 0.88. Average expected morbidity was 43.8%, expected cases were 116. Observed morbidity rate was 39.6% (105/265). The expected and observed morbidities and cases had no significantly differences. All patients were classified to 1 of 4 strata based on their individual POSSUM scores and subsequent risk of morbidity. Predictive value was the highest when scores ranged from 0.4 to 0.8. POSSUM exhibited less predictive value for mortality, but if POSSUM was more than 0.5, it was useful for mortality predicting.</p><p><b>CONCLUSIONS</b>POSSUM scoring system has high value for predicting the risk of morbidity in PD and can be helpful in guiding surgery and postoperative management decisions.</p>

Secretion expression of cholera toxin B subunit in food-grading Lactococcus lactis expression system / 中华流行病学杂志

<p><b>OBJECTIVE</b>To clone and secretion express cholera toxin B subunit (CTB) in food-grading Lactococcus lactis expression systems.</p><p><b>METHODS</b>ctB fragment that encoding CTB was amplified by polymerase chain reaction (PCR) using the genomic DNA of Vibrio cholera strain 569B as template and was inserted into two secretion expression vector pSQZ and pSQ to construct food-grading expression system L.lactis MBP71/pSQZ-ctB and L.lactis MBP71/pSQ-ctB. The expressed CTB was detected by Western-blot assay.</p><p><b>RESULTS</b>The ctB fragment was successfully amplified from Vibrio cholera strain 569B and inserted into two secretion expression vectors pSQZ and pSQ to construct food-grading expression system L. lactis MBP71/pSQZ-ctB and L. lactis MBP71/pSQ-ctB. Western-blot assay demonstrated that CTB was secretion and expressed from L.lactis MBP71 harboring vectors pSQZ-ctB and pSQ-ctB, and the quantity of CTB secreted by L. lactis MBP71/pSQ-ctB was about 2 microg/ml, higher than that of L. lactis MBP71/pSQZ-ctB.</p><p><b>CONCLUSION</b>CTB was successfully secreted and expressed by food-grading L. lactis expression systems.</p>

Molecular typing of isolates from two dysentery outbreaks in Chengdu, through pulsed field gel electrophoresis technology / 中华流行病学杂志

<p><b>OBJECTIVE</b>Using pulsed field gel electrophoresis (PFGE) and polymerase chain reaction (PCR) typing to analyze strains isolated from two outbreaks caused by Shigella sonnei and to trace the source of infection.</p><p><b>METHODS</b>Virulence genes ipaH and ial were detected by PCR and PFGE was used to subtype the isolates. Patterns were compared, using the software BioNumerics.</p><p><b>RESULTS</b>Within the 54 isolates, all were ipaH positive with 48 as ial positive. Strains from the Chongzhou outbreak were clustered into 4 PFGE patterns, with the predominant pattern accounted for 72% of the analyzed strains. The pattern of strains isolated from the cold pork with sauce was identical to the predominant pattern. The strains from Dayi outbreak were clustered into 8 PFGE patterns and the predominant pattern accounted for 56% of the test strains.</p><p><b>CONCLUSION</b>Strains from the two outbreaks were quite different and the 'cold pork with sauce' seemed to be the major source of infection, causing the outbreak of diarrhea in Chongzhou. The sources of infection of the Dayi outbreak might be complicated whereas PFGE showed a discriminatory and reproducible laboratory tool in the epidemiologic investigation on outbreaks of diarrhea.</p>

Clone and express ctb-stx2 b fusion gene in Enterohemrrhagic escherichia coli O157:H7 Shigeai toxin 2B subunit and V cholera toxin B subunit and the detection of their immunogenicity / 中华流行病学杂志

Objective To clone and express the fusion gene encoding Enterohemrrhagic escherichia coli O157:H7(EHEC O157:H7)Shigela toxin 2B subunit(Stx2B)and vibrio cholera toxin B subunit (CTB)as well as to detect the immunogenicity and GM1-binding ability of fusion protein.Methods To design a primer to amplify stx2b gene and ctb-stx2b fusion gene encoding Stx2B and CTB-Stx2B respectively and to clone the genes into express plasmid pET30a(+)C in order to construct pET30a-ctb-stx2b after T-A sequencing was varified,then to transform constructed plasmid into E.coliBL21(DE3)induced by IPTG and purified by a purify kit and to detect molecular weight and immunogenicity by SDSPAGE and Western-blot.Results The amplified ctb-stx2b fragments appeared to he 750 bp and gene sequence was identical to designed sequence.The prokaryotic expression system pET30a-ctb-stx2b/BL21 could express protein weight about Mr20×103and the expressed protein could react to CTB monoclone anti-body.The fusion protein CTB-Stx2B could bind GM1.Conclusion CTB-Stx2B had successfully been expressed in prokaryotic while the expressed protein had good immunogenicity and GM1-Binding ability.This study provided information on further EHEC O157:H7 vaccine research.

Analysis on the status of Shigella spp antimicrobial resistance through data from the National Shigellosis Surveillance System in China, in 2005 / 中华流行病学杂志

<p><b>OBJECTIVE</b>The Ministry of Public Health released the National Surveillance project on Shigellosis in August, 2005. This study was to reveal the antimicrobial resistance status of Shigella isolates through the National Shigellosis Surveillance System in 2005 in China, so as to provide evidence for the development of surveillance, prevention and cure of Shigellosis.</p><p><b>METHODS</b>All the lab assistants received training from Chinese Center for Disease Control and Prevention. The project prescribed the uniform experimentation, quality control method, reagent, etc. Disc diffusion test(K-B) was carried out, following the CLSI methods. Data were analyzed by WHONET 5.4 software.</p><p><b>RESULTS</b>(1) 3 serotypes were identified and S. flexneri was common that accounted for 75.5% of all Shigella isolates followed by 24.4% of S. sonnei, but only 1 strain of S. dysenteriae was separated. (2) The resistant rates to tetracycline and ampicillin in Shigella spp were quite high, as over 90.0%. However, the resistant rate to Cefotaxime was the lowest, only 6.1%. The resistant rates were different between serotypes with the resistant rates of S. flexneri to ampicillin, ampicillin/clavulanate and ciprofloxacin were higher than those of S. sonnei (P < 0.001). (3) The multiple-antibiotic-resistance status in Shigella spp was quite serious and the resistant rate to five and more antimicrobials was 54.9%. The most common resistant patterns were seen on ampicillin, nalidixin, tetracycline and sulfamethoxazole. (4) There were some differences in subtypes and antimicrobial resistance among different provinces.</p><p><b>CONCLUSION</b>Cefotaxime seemed the best in curing Shigellosis at the clinic level. Programs regarding monitoring subtypes and antimicrobial resistance of Shigella should be in a continuous manner so as to understand the pathogens timely and to control the disease pertinently.</p>

Surveillance on severe acute respiratory syndrome associated coronavirus in animals at a live animal market of Guangzhou in 2004 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the prevalence of severe acute respiratory syndrome coronavirus (SARS-CoV) like virus in animals at a live animal market of Guanzhou in 2004 before and after culling of wild animal action taken by the local authority, in order to predict the re-emerging of SARS from animal originals in this region.</p><p><b>METHODS</b>Animals at live animal market were sampled for rectal and throat swabs in triplicate. A single step realtime reverse transcription-polymerase chain reaction (RT-PCR) diagnostic kit was performed for screening SARS-CoV like virus, the manual nested RT- PCR and DNA sequencing were performed for confirmation. Only specimens which tested positive for both of the N and P genes by nested RT-PCR were scored as positive.</p><p><b>RESULTS</b>In 31 animals sampled in January 5 2004 before culling of wild animals at Guangdong Province, including 20 cats (Felis catus), 5 red fox (Vulpes vulpes) and 6 Lesser rice field rats (Rattus losea), 8 (25.8%) animals were tested positive for SARS-CoV like virus by RT-PCR methods, of which 4 cats, 3 red fox and one Lesser rice field rats were included. However, two weeks after culling of animals and disinfection of the market were implemented, in 119 animals sampled in January 20 2004, including 6 rabbits (Oryctolagus cuniculus), 13 cats, 46 red jungle fowl (Gallus gallus), 13 spotbill duck (Anas platyrhynchos), 10 greylag goose (Anser anser), 31 Chinese francolin (Franclinus pintadeanus), only rectal swab from one greylag goose was tested positive for SARS-CoV like virus. Furthermore, in 102 animals that including 14 greylag gooses, 3 cats, 5 rabbits, 9 spotbill duck (Anaspoecilorhyncha), 2 Chinese francolin (Franclinus pintadeanus), 8 common pheasant (Phasianus colchicus), 6 pigeons, 9 Chinese muntjac (Muntiacus reevesi), 19 wild boar (Sus scrofa), 16 Lesser rice field rats, 5 dogs, 1 mink (Mustela vison), 3 goats, 2 green peafowl (Pavo muticus) sampled in April, May, June, July, August and November, only rectal swab from one pig was tested positive. However, of 12 and 10 palm civets sampled in November and December including five of which had been at the live animals market for 2 days, none of them was tested positive.</p><p><b>CONCLUSION</b>This findings revealed that animals being sampled in April, May, June, July, August and November of 2004, only one rectal swab from a pig was tested positive as SARS-CoV like virus, much lower than the results from the previous year, suggesting that the possibility of re-emerging of human infection from animal origins is low for the winter of 2004-2005.</p>