Objective To determine Haemophilus influenzae type b strains in molecular level using PCR,and to study the immunogenicity of capsular polysaccharide conjugates in mice.Methods Extracting genomes using bacterial DNA extract kit from Hoemophilus influenzae type b strains,and PCR for determining the strains through serotyping-specific and capsular genotyping primers respectively.Various capsular polysaccharides conjugated TT respectively,and the conjugates were administered subcutaneously to mice through dilution.After vaccination with two doses,blood samples were collected for the detection of antibody levels to polyribosylribitol phosphate ( PRP),the capsular polysaccharide of Hib.Results All five Haemophilus influenzae type b strains contain type-specific(482 bp) and capsular type (343 bp)DNA fragment through PCR detecting.The DNA fragments were sequenced.BLAST show that these sequences are 100％ homology comparing the above strains respectively,and are 99％ and 100％ homology comparing the GenBank X78559.1 and M19995.1 respectively.The immunogenicity of mice from various capsular polysaccharide conjugates (PRP-TT) was not significantly different by ELISA detecting.Conclusion Through PCR,Haemophilus influenzae type b strain can be determined in molecular level.The immunogenicity of mice from purified capsular polysaccharide conjugates was not different.The study provides a detection means for the features and heredity stability of Haemophilus influenzae type b strain and reference data for the immunogenicity of different polysaccharide conjugates in vaccine research and development and production.

Objective · To compare the prevalence of tobacco advertisement before and after implementation of the new edition of Advertising Law of the People's Republic of China and evaluate the effects of implementation of the new Advertising Law. Methods · Tobacco sales places, public places, and streets in Shanghai were randomly selected to observe tobacco advertisement and complete the survey. The survey was carried out in 10 districts of Shanghai in June 2015 with 10 tobacco sales places selected in each district, and in 4 districts of Shanghai in June 2016 with 25 tobacco sales places selected in each district. Sales places included convenient stores, tobacco stores, supermarkets, and kiosks. In addition, public transport waiting points, public transport vehicles, hospitals, shopping centers, restaurants, bars, and Internet cafes were selected into survey in 2016. Results · In 2016, the proportion of tobacco sales places having tobacco advertisement was lower than that in 2015, but the proportion of tobacco promotion behaviors was higher. In 2016, 128 public places and streets were investigated and only 1 had outdoor tobacco advertisement. Conclusion · The new edition of Advertising Law has significant effect on controlling tobacco advertisement, but tobacco sales places still have tobacco advertisement with various disguised promotion forms. Tobacco sales places have diversified business styles and minors often enter these places. It is recommended that a clear definition of disguised forms of tobacco advertisement should be made, tobacco advertisement at tobacco sales places needs to be supervised, the release of tobacco advertisement should be strictly reviewed, and existing tobacco advertisement should be severely punished, so as to prevent minors from exposing to tobacco advertisement and information luring smoking.

Objective To understand pathogen spectrum of bacterial and fungal infection of central nervous system (CNS),and evaluate the etiological diagnostic value of universal primer polymerase chain reaction (PCR).Methods Data about patients with suspected or confirmed bacterial and fungal infection of CNS from January 2009 to March 2015 were collected,species of pathogens from cerebrospinal fluid (CSF)were analyzed,DNA from patients’CSF were performed PCR amplification and sequencing with universal primers of bacterial 16S rRNA and fungal 28S rRNA, PCR detection results were compared with CSF culture during the same period.Results A total of 400 patients were with confirmed or suspected bacterial or fungal infection of CNS,132 of whom were with positive CSF culture.150 pathogenic isolates were detected,including 48 isolates of gram-positive bacteria,90 gram-negative bacteria,and 12 fungi;the top three isolated bacteria were Acinetobacter baumannii (n =32 ),coagulase negative staphylococcus (n=16)and Klebsiella pneumoniae (n=13);the most common fungus was Cryptococcus neoformans (n=8).CSF from 88 infected patients and 20 non-infected patients were selected for PCR amplification,the sensitive of PCR am-plification assay was higher than the culture method (35.23% [31/88]vs 28.41 %[25/88],χ2 =4.17,P <0.05).

Objective To evaluate the value of amplification and sequencing of 16S rRNA gene in the identification of clinical rare pathogenic bacteria,and guide the diagnosis and treatment for related clinical infection.Methods 12 bacterial isolates that were difficult,or unable to be identified with conventional laboratory methods,or with special phenotypes were collected. The 16S rRNA gene was amplified by polymerase chain reaction (PCR),then sequenced for identifying bacterial species through BLAST comparison,clinical characteristics of related infection were ana-lyzed.Results 12 clinical isolates were all positive for PCR amplification (about 1500 bp),species were all identi-fied (similarity≥99% ),the identified strains were Listeriamonocytogenes(n= 2),Brucellamelitensis(n= 2),Fu-sobacteriummortiferum(n= 1),Rothiaaeria(n= 1),Nocardiafarcinica(n= 1),Staphylococcussaccharolyticus (n= 1 ),Rhizobiumradiobacter(n= 1 ),Prevotellabivia(n= 1 ),Ralstoniamannitolilytica(n= 1 ),and Atopobium vaginae(n= 1 ). The sensitivity of 16S rRNA gene amplification was high,and the minimum detection limit of Escherichiacoli ATCC 25922 was 1.5×101 CFU/mL. Clinical data of 12 patients revealed that these strains can cause multi-sites and multi-types of infection,after patients received targeted antimicrobial therapy,11 improved, and 1 died.Conclusion Sequencing for 16S rRNA gene can rapidly and accurately identify rare,anaerobic,and difficult cultured bacteria,provide laboratory evidence for etiological diagnosis and treatment of different types of infection.

Objective To investigate the immunogenicity of group A plus group C meningococeal glycoconjugate vaccine,namely dosage,immune memory and compatibility. Methods The mice were injected with group A plus group C meningococcal glycoconjugate vaccine with different dosages. Blood samples were taken on the 14th day after the last injection for testing the antibodies against polysaccharide A and C. After the optimal immunization dosage had been decided,the mice were inoculated separately with the monovalent group A and the monovalent group C and the bivalent group A plus group C glycoconjugate vaccine with one,two or three injections for observation of the effectiveness of different injections and the compatibilities. Results The dosage of 1.25 μg of each polysaccharide of group A and group C in bivalent glycoconjugate vaccine appears to be immunologically optimal to vaccinate the mice. Immunological memory could be induced in mice inoculated with the glycoconjugate vaccine,and the antigenic immunogenicity of the group A component and group C component in the formulation of group A plus group C meningococcal glycoconjugate vaccine was not affected. Conclusion Group A plus group C meningococcal glycoconjugate vaccine have good immunogenicity,immune memory and compatibility.

Objective To examine the clinical value of polymerase chain reaction (PCR) in rapid diagnosis of bacterial and fungal infection of central nervous system.Methods The cerebrospinal fluid (CSF) samples were collected from 137 patients for DNA extraction.PCR was used to amplify the DNA of pathogenic bacteria and fungi using universal primers.The PCR products were subjected to DNA sequencing analysis for identifying microbial species.The conventional culture of pathogens was carried out simultaneously as control.Results PCR revealed bacterial pathogen in 50 of the 137 CSF samples,fungal pathogen in 6 of the 137 CSF samples.Conventional culture of CSF reported positive bacterial infection in 38 cases,fungal infection in 5 cases.PCR provided diagnostic sensitivity of 40.9％,specificity 100％,positive predictive value 100％,negative predictive value 38.2％.The diagnostic efficiency was 56.7％.In contrast,the conventional culture achieved the results of 31.4％,100％,100％,34.7％,44.4％,respectively.The sensitivity,negative predictive value,and diagnostic efficiency of PCR were significantly better than conventional culture method.The coincidence rate between PCR and conventional culture was 97.7％.Conclusions Universal primer-based PCR is characteristic of short turnaround time,specificity,sensitivity and accuracy,which is very useful for rapid diagnosis of the pathogenic bacteria and fungi in central nervous system infections.

Objective:To investigate the independent risk factors for fever after endoscopic radiofrequency ablation (RFA).Methods:From January 2016 to April 2021, 51 patients with early esophageal cancer, who were treated with RFA in the Department of Gastroenterology, Changhai Hospital and whose lesion range exceeded 3/4 of the circumference of esophagus, were included in the case-control study. Patients were divided into fever group ( n=15) and non-fever group ( n=36) according to whether they had fever after operation. The general condition of patients, family history of gastrointestinal tumors, lesion length, lesion range, ablation energy and ablation times were mainly collected for univariate analysis. The variables with P<0.1 were further included in multivariate logistic regression analysis to explore the independent risk factors for fever after RFA. Results:Univariate analysis showed that the lesion length ( t=-3.89, P<0.001), lesion range ( χ2=11.52, P=0.001) and ablation energy ( P=0.001) were significantly different between the two groups. Pearson correlation showed that there was a significant positive correlation between lesion length and lesion circumference ( r=0.71, P<0.001), and the lesion range was determined by the lesion circumference length. Therefore, the two variables of lesion length and ablation energy were finally included in the logistic regression analysis. Logistic regression analysis showed that the risk of fever after RFA was 1.21 times as high as that before when the length of esophageal lesions increased by 1 centimeter (95% CI: 1.01-1.43, P=0.037). The risk of fever after RFA using 12 J ablation energy was 0.43 times as high as that using 10 J ablation energy (95% CI: 0.22-0.85, P=0.015). Conclusion:Lesion length and ablation energy are independent risk factors for fever after esophageal RFA. Patients with long segment early esophageal cancer and using low ablation energy are more likely to have fever after RFA.

Objective: To investigate the influencing factors of smoking tendency of adolescents and to analyze the influence of e-cigarette on smoking tendency of teenagers and the possible interaction, to provide evidence for tobacco control measures. Methods: Stratified multistage cluster probability sampling method was used to select 6 178 students from junior middle school, high school and vocational high school students in Shanghai for questionnaire survey from September to October 2019. Students with smoking tendency were taken as case group, adolescents with same sex and similar age(within 1 year) were mathed in 1∶3 ratio. A total of 631 adolescents with smoking tendency and 1 870 non-smoking tendency were included in the study. Conditional Logistic regression was used to analyze the influencing factors and possible interactions. Results: After adjusting gender, age and other covariates, conditional Logistic analysis results showed that the smoking tendency of middle school students was associated with factors such as their friends smoking, trying cigarettes and trying e-cigarettes. The OR value and 95%CI were 3.26(2.47-4.29), 5.90(3.76-9.24), and 3.28(2.11-5.10), respectively. The interaction analysis results showed that the OR value and 95%CI of the multiplying interaction between friends smoking and trying e-cigarettes, trying smoking and trying e-cigarettes were 8.62(4.90-15.17) and 12.01(6.02-23.95), respectively. There was no additive interaction. Conclusion Tobacco control interventions, such as e-cigarette harm publicity and peer education, can help to change teenagers smoking tendency and further reduce their tobacco use rate.

Objective To investigate the molecular epidemiology and clinical characteristics of human bocavirus in children with acute respiratory tract infection in Shanghai,China.Methods Between January 2012 and December 2012,271 nasopharyngeal aspiration (NPA) samples were collected from children who had been hospitalized for acute respiratory tract infection at Shanghai,China.HBoV NS1 gene was detected by the nested polymerase chain reaction.In the further,other common respiratory viruses (HRV,RSV,ADV et al) was screened in HBoV positive samples,All PCR positive products were sequenced and phylogenetic analysis.Results The overall frequency of HBoV,infection was 11.4％.21of 31 HBoV positive sample tested were mix-infection.HBoV was detected from all over year.Children with HBoV varied in age from 4 months to 4-years (median age,17 months).The clinical diagnoses of HBoV positive patients included acute upper respiratory tract infection and lower respiratory tract infection.The clinical presentations of HBoV positive children included fever,cough,sputum production,diarrhea,vomiting;pharynx engorgement,crackles,wheezing.There being a statistically significant difference in the detection rates of HBoV between the outpatients and inpatients.The HBoV NS1 gene sequences phylogenetic analysis revealed that 29 HBoV1 NS1 sequences shared 99％-100％ nucleotide identity with the human bocavirus strain,whereas the amino acid identity was 96％-100％.The two HBoV2 NS1 sequences shared 99％ nucleotide sequence identity with HBoV2 strain FJ170282 respectively.Conclusions Human bocavirus was distributed in Shanghai region,HBoV1 was dominant and HBoV2 was detected at the first time.The symptom and clinical diagnoses has no specificity with patients with other common respiratory viruses.

Objective: To investigate the effect of treatment dentures on changes in denture space in edentulous patients and to evaluate its clinical effect. Methods: Twenty patients with treatment dentures were investigated with a questionnaire and Oral Health Impact Profile (OHIP-14) before and after treatment. The denture space, denture tissue surface, occlusal points, Gothic arch images, mucosal condition, lateral occlusal condition, bite force, retention and stability, patient mastication ability, denture satisfaction, and OHIP-14 score were collected for statistical analysis. Results : After wearing treatment dentures, the denture space significantly improved, the tissue conditioners on the treatment denture gradually began to be distributed, and the occlusal points gradually became symmetrical. The gothic arch images showed that the joints, nerves, and muscles gradually stabilized. The mucosal condition changed to a healthy state. The lateral occlusal conditions were improved. There were also significant differences in the bite force, retention and stability, subjective and objective chewing ability, satisfaction with the denture and OHIP-14 score before and after treatment (P < 0.05). The objective chewing ability was positively correlated with the retention stability of the denture, the subjective chewing ability and the satisfaction of the denture, and it was negatively correlated with OHIP-14. Conclusion Treatment dentures could help to restore denture space to an ideal condition and improve the oral health of patients. This treatment could also improve the retention and stability of the denture and the chewing ability to improve patient satisfaction. This treatment has clinical application value and promotion potential.