Objective To investigate the correlation between CD40 gene promoter region - 1C/T polymorphism and carotid atherosclerotic plaques and large artery atherosclerotic stroke.Methods The subjects were the patients with acute large artery atherosclerotic stroke (patient group) and the patients without history of stroke (control group).The patient group was further divided into an unstable plaque subgroup,a stable plaque subgroup,and a plaque-free subgroup.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect CD40 - 1C/T polymorphism.Results A total of 170 patients with large artery atherosclerotic stroke (patient group) and 61 subjects without history of stroke (control group) were included.In the patient group,51 patients were in the unstable plaque subgroup,60 were in the stable plaque subgroup,and 59 were in the plaque-free subgroup.C allele frequency of the patient group was significantly higher than that of the comrol group (52.9％ vs.38.5％ ;x2 =7.466,P =0.006).In the patient group,C allele frequency of the unstable plaque subgroup (75.5％) was significantly higher than that of the stable plaque subgroup (53.3％),and both of them were significantly higher than that of the plaque-free subgroup (33.1％ ) (the stable plaque subgroup vs.the plaque-free subgroup:x2 =9.970,P =0.002; the unstable plaque subgroup vs.the stable plaque subgroups:x2 =11.680,P =0.001; the unstable plaque subgroup vs.the plaque-free subgroup:x2 =39.532,P=0.000).Multivariate logistic regressive analysis showed that hypertension (OR9.513,95％ CI 1.291 - 20.779; P =0.028),increased total cholesterol level (OR 4.235,95％ CI 1.069 -19.034; P =0.032),increased low density lipoprotein level( OR 4.201,95％ CI 1.803 - 9.672; P =0.001 )and C alleles (OR 1.759,95％ CI 1.177 - 2.738; P =0.006) are the independent risk factors of large artery atherosclerotic stroke.Conclusions CD40 - 1C/T polymorphism is associated with the risks of carotid atherosclerotic plaque formation,unstable plaque and large artery atherosclerotic stroke; C allele may be a susceptibility factor for large artery atherosclerotic stroke.

ObjectiveTo discuss the expression of tumor shift suppressor gene KAI1,matrix metal proteinase 9 (MMP-9) in endometriosis sickness in dystopia internal membrane and its significance.MethodsIn 53 patients and 30 healthy controls,the expression of KAI1,MMP-9 in the endometriosis was detected with immunohistochemistry SP method,and the relationship was analyzed.ResultsKAI1 had low expression in EMS patients,while MMP-9 was highly expressed.In the control group,KAI1 was highly expressed while MMP-9 lowly expressed,and the differences between two groups were remarkably significant (P ＜0.05).KAI1 and the MMP-9 expression showed negative correlation between two groups.Conclusions KAI1 and MMP-9 was negatively expressed in EMS,which may play a role in the EMS prognosis.KAI1 and MMP-9 may be served as the prognosis and monitor markers for EMS.

Objective To investigate the mucosal immunity of L1 virus-like particles ( VLPs) of human papillomavirus ( HPV) types 16 and 18 plus JY adjuvant by intranasal immunization in cynomolgus. Methods Cynomolgus were immunized with low and high dosage of HPV types 16 and 18 L1 VLP with JY adjuvant for 3 times by intranasal route at weeks 0, 4 and 8, respectively, using PBS as control. Subsequently, vaginal secretion, oral secretion and nasal secretion were collected at weeks 0, 2, 4, 6, 8 and 16, respectively, and determined for mucosal immunity by ELISA.Results HPV-L1-specific sIgA antibodies were detected in all secretions, including oral, nasal and vaginal ones, the concentrations of sIgA antibody induced were much higher than those in PBS control group.There was no significant difference ( P>0.05) in sIgA antibody levels among cynomolgus vaccinated with low and high dosage of L1 VLP, as was between oral and nasal secretion ( P >0.05 ) , However, the concentrations of sIgA antibody in vaginal secretion were significant higher than those in oral and nasal secretion, differences were statistically significant ( P<0.01) .Conclusions Following intranasal immunization in cynomolgus, HPV types 16 and 18 L1 VLP with JY adjuvant can effectively induce sIgA antibody in vaginal secretion, and vaginal sIgA antibody concentrations were much higher than those in oral and nasal secretion.

Objective To evaluate the effect of JY adjuvant,which is composed of IL-2 and chitosan,on the immune response for mucosal immunization with human papillomavirus types 16 and 18 L1 virus-like particles (HPV16 + 18 L1 VLP).Methods Mice were immunized three times with HPV16 + 18 L1 VLP in the presence or absence of JY adjuvant by intramuscular and intranasal routes,respectively.Subsequently,experiments were undertaken to detect serum IgG antibody,neutralizing antibody and respiratory tract washes sIgA antibody titers and cellular immune response.Results Following intranasal immunization,serum IgG antibody titers were much higher in HPV16 + 18 L1 VLP with JY adjuvant group than that in VLP without adjuvant group (P ＜ 0.01),after intramuscular immunization,serum IgG titers induced by VLP with or without JY adjuvant were the same; following intramuscular immunization,neutralizing antibody titers induced by adjuvant-containing HPV16 + 18 L1 VLP were higher than those by adjuvant-free VLP,following intranasal immunization,only serum neutralizing antibody was detected in adjuvant-containing VLP group; after intranasal immunization,lung washes sIgA concentration were much higher in HPV16 + 18 L1 VLP with JY adjuvant group than that in VLP without adjuvant group (P ＜ 0.05) ; following intranasal and intramuscular immunization,respectively,the number of spot forming cells were much higher in HPV16 + 18 L1 VLP with JY adjuvant group than that in VLP without adjuvant group (P ＜ 0.01,P ＜ 0.05,respectively).Conclusion JY adjuvant enhanced the cellular,humoral and mocosal immunities induced with HPV16 + 18 L1 VLP by intranasal route,while showed no significant influence of the adjuvant was seen in the group immunized by intramuscular route.

Objective We develop a rapid,specific,sensitive tandardized SOP.And initial application for 200 nasopharyngeal aspirates of children with related pathogens of acute respiratory tract infections in BeiJing area.Methods To developed nested PCR and TaqMan probe real-time fluorescence quantitative PCR method for detection of KIPyV and WUPyV'S gene,and then the sequences of gene fragments are analyzed.Evalution of two assays from 200 nasopharyngeal aspirates.Results In this study,sensitivity of TaqMan probe real time fluorescent quantitative PCR assay was higher than one of nested PCR (500 copies/μl),and both assays did not show any positive amplification in detetion of other respiratory virus.Coefficient of varience of KIPyV and WUPyV are less than 2.9％ and 1.95％ respectively in the repeatability detection.The detection rates of KIPyV,and WUPyV were 1.5％ and 8％ in nested PCR assay and 12％ and 14％ in real time Fluorescent quantitative PCR assay respectively.Conclusion This study established good sensitivity and reproducibility,high specificity and rapid method for detection nucleic acid of these polyomaviruses that have good prospects on the clinical application.

Objective:To retrospectively analyze the clinical characteristics of elderly patients who received colonoscopy and to explore the clinical value of regular colonoscopy for the elderly.Methods:This was a retrospective cohort study.A total of 1 154 patients aged 75 years and over undergone colonoscopy in Guangdong General Hospital from January 2015 to March 2018 were enrolled and divided into three groups, including 605 cases aged 75-79 years, 527 cases aged 80-89 years and 22 cases aged 90 years and over.Detection rates of colorectal lesions by colonoscopy were recorded.The clinical value of annual colonoscopy on the detection of colorectal lesions in elderly patients 75 years and older were analyzed to assess the necessity for regular monitoring.Results:Overall, 569 cases(49.3%)underwent colonoscopy with sedation and 585 cases(50.7%)underwent colonoscopy without sedation.The total positive detection rate was 83.4%(962/1 154), and the main lesions were polyps(858 cases, 74.4%), including 605(52.4%)cases of adenomas.Among the three groups, gastrointestinal bleeding was the main cause for colonoscopy in the group aged 90 years and over, while abdominal discomfort, elevated immunological tumor markers and history of non-colon cancer were the main reasons for colonoscopy in the group aged 75-79 years( P<0.05). A total of 153 cases underwent annual colonoscopy.The detection rate of polyps and adenomas decreased in the second exam, but still higher than 40.0%. Conclusions:Colonoscopy is a safe and effective method for the elderly population aged 75 years and over.Polyps and adenomas are the most common lesions.Recurrence of polyps after colorectal cancer and polypectomy is common and it is necessary to receive colonoscopy regularly.

Objective:To study the sequence characteristics and genetic variation of a human respiratory syncytial virus (HRSV) subtype A genome in Beijing.Methods:The genomic RNA of HRSV from nasopharyngeal aspirate samples was sequenced and obtained a whole genome sequence of HRSV A subtype. The phylogenetic tree was constructed with reference sequences of other HRSV strains. The major proteins were compared and single nucleotide polymorphism analyzed. In addition, the N-glycosylationsites of F and G protein were predicted.Results:Phylogenetic tree and homology analysis results suggest that the HRSV strain (RSVA/Beijing-China/2017) was the A subtype ON1 genotype. Nucleotide and amino acid variation analysis showed that G protein, F protein and L protein had some substitutions. Analysis of amino acid variation sites showed that amino acid substitution (L142S) occurred at position 142 of G protein. For F protein, there were two substitutions, which were S105N in the P27 peptide (110-136aa) and C69Y in the antigen sites ? (62-69 aa and 196-210 aa). The prediction of N-glycosylation sites revealed that there were 5 N-glycosylation sites of F protein and 4 N-glycosylation sites of G protein in this strain.Conclusions:The HRSV strain obtained in Beijing belongs to A subtype ON1 genotype. The G, F and L proteins have large variations, and 22 amino acid substitutions have occurred in the G and F proteins.

Objective:To investigate the epidemiological characteristics of human bocavirus 1 (HBoV1) and to analyze the genetic variation.Methods:A total of 2 848 nasopharyngeal aspirate (NPAs) specimens were collected from hospitalized children with acute respiratory tract infections (ARTI) in Beijing Friendship Hospital from April 2017 to March 2019, and HBoV1 was detected by quantitative real-time PCR. Epidemiological analysis was carried out based on the clinical information of the patients. The nested PCR method was used to amplify the NP1 and VP1 genes of HBoV1 for homology analysis. Maximum clade credibility tree (MCC tree) and genetic polymorphism map were constructed to analyze the time evolution of HBoV1 VP1.Results:HBoV1 was detected in 90(3.16%) of 2 848 NPAs, most (93.33%, 84/90) HBoV1-positive cases were among children <5 years of age. HBoV1 could be detected throughout the year with a higher prevalence 7.23% (18/249) in October. Of the 90 HBoV1-infected cases, the main clinical symptoms were fever and cough, 44(48.89%) were co-infected with other respiratory viruses; 55 NP1 sequences and 47 VP1 sequences were obtained by nested PCR amplification, phylogenetic analysis showed that the nucleotide homology was 98.9%~-100% and 99.1%~-100%, respectively. MCC tree showed that the HBoV1 VP1 gene sequence obtained in this study appeared in two adjacent clades, the gene evolution was stable.Conclusions:HBoV1 is one of the common viruses that cause respiratory infection among children in Beijing. HBoV1 genetic evolution is relatively stable, but it still needs to be monitored continuously.

Objective:To construct 2019 novel coronavirus (2019-nCoV) 614D and 614G pseudovirus by HIV lentivirus packaging system and explore their biological specificity.Methods:The recombinant expression plasmids pCDNA3.1-614D and pCDNA3.1-614G were transiently cotransfected with psPAX2 and pLenti CMV Puro LUC into 293T cells respectively. After 72 hours, the supernatant was collected and ultracentrifuged with 20% sucrose cushion. The titer, morphology, protein expression and neutralizing activity of pseudovirus were determined.Results:S protein specific fluorescence was detected by indirect immunofluorescence test, Western blot analysis showed S protein was expressed, and the spike of pseudovirus was observed under transmission electron microscope. The titers of pseudovirus 614D and 614G were 1.12×10 4 and 2.52×10 4 TCID 50/ml, respectively. The pseudovirus 614D and 614G could be neutralized by S rabbit polyclonal antibody, indicating that the pseudovirus has high specificity. Conclusions:In this study, 2019-nCoV 614D and 614G pseudovirus was successfully constructed, which laid the foundation for the establishment of in vitro neutralizing antibody detection platform based on pseudovirus.

Objective To analyze the distribution characteristics of injuries among students in Songjiang district, Shanghai, and to provide data support for preventing injuries among students. Methods Data of injury cases among school students in Songjiang from 2017 to 2019 were collected from the National Injury Surveillance System (NISS), and the demographic characteristics, injury occurrence and clinical characteristics of injuries were analyzed. Results A total of 13 877 cases of injuries in school students were reported from 2017-2019, with a sex ratio of 1.94:1. The education level of the injured students was mainly primary school. The peak incidence months of student injuries were May (9.97%) and November (9.67%). Falls (50.83%) were the leading cause of student injuries, and the top three places of student injuries were at home (30.08%), in schools and public places (28.11%) and on roads/streets (22.97%). The top three activities at the time of the injury were leisure activities (34.62%), sports activities (18.50%), and taking transportation（17.68%）. The top three injury types were bruise/scratch (42.94%), sprain/strain (28.67%) and sharps/bite/open injury (13.53%). The common injury parts were lower limbs (38.39%), head (27.82%) and upper limbs (24.88%). Conclusion The prevention of injuries in school students in Songjiang should focus on male students and low age students, and targeted injury prevention and control work for students of different ages should be carried out.