ObjectiveTo analyze the epidemiological characteristics of 8 major respiratory pathogens in influenza-like illness (ILI) cases with acute respiratory infections at fever clinics in Huangpu District, Shanghai from 2015 to 2024, and to provide a scientific basis for the prevention and treatment of respiratory diseases. MethodsA retrospective study was conducted in Huangpu District. Individuals meeting the case definition of ILI from 2015 to 2024 was registered. Their nasopharyngeal swabs were collected for pathogen detection. A total of 8 respiratory viruses were tested, including Influenza A virus (Flu A), Influenza B virus (Flu B), adenovirus (ADV), enterovirus/human rhinovirus (EV/HRV), human parainfluenza virus (HPIV), human coronavirus (HCoV), respiratory syncytial virus (RSV), and human metapneumovirus (HMPV). ResultsFrom 2015 to 2019, a total of 344 ILI cases were tested, of which 192 out of 344 cases (55.81%) were tested positive for single respiratory pathogen. From 2023 to 2024, 1 557 ILI cases were tested, with 572 out of 1 557 cases (36.74%) being positive for single pathogen. From 2023 to 2024, the positive rate of single pathogen in ILI cases was significantly lower than that in 2015‒2019 (χ²=42.66, P<0.001). Specifically, the positive rate of Flu A (χ²=74.43, P<0.001) decreased, while that of HPIV (χ²=8.66, P=0.003) increased, both with statistically significant differences. According to the seasonal pattern, the epidemic intensity of Flu A decreased in summer, while that of HPIV increased in summer and autumn. Demographic results showed statistically significant differences in the positive rates of EV/HRV between genders (χ²=22.38, P<0.001), with males exhibiting a higher positive rate than females. No statistically significant differences were identified in the positive rates of single pathogen among different age groups (χ²=4.42, P=0.110). Nevertheless, statistically significant differences were noted when comparing the positive rates of EV/HRV, Flu A, Flu B and HPIV across different age groups (P<0.05). EV/HRV was more commonly detected in the 15‒<25 age group (10.93%), while Flu A and HPIV had the highest positive rates in the ≥60 age group (21.24% and 4.77%). Flu B had the highest positive rate in the 25‒<60 age group (11.26%). 52.63% of cases with co-infections occurred during winter, with the primary pathogens involved being EV/HRV (9 cases) and HCoV (6 cases). The most prevalent combination of co-infection was Flu A with EV/HRV. ConclusionThe prevalence of respiratory pathogens among ILI cases from 2023 to 2024 exhibited notable fluctuations compared to that from 2015 to 2019. Therefore, influenza surveillance should be strengthened, and attention should also be paid to the prevalence of respiratory pathogens such as HPIV. These findings have profound implications for future research, surveillance, vaccine planning, and public health policy making.

Objective:To investigate the impact of inserting an exogenous gene, NanoLuc (Nluc), at different sites in the influenza virus genome on viral properties and analyze the expression stability of the exogenous gene both in vitro and in vivo. Methods:Using molecular cloning techniques and reverse genetics, eight recombinant influenza viruses were constructed by inserting the exogenous Nluc gene into the gene segments encoding hemagglutinin (HA), neuraminidase (NA), non-structural protein (NS), and polymerase basic protein 1 (PB1). Viral replication capacity was evaluated by hemagglutination and plaque assays. Nluc expression in infected cells was monitored by fluorescence imaging. The potential impact of the exogenous gene insertion on viral infectivity was examined in a mouse infection model. Independent samples t-test were used for statistical analysis. Results:The recombinant viruses with insertions in the HA, NA, and NS gene segments generated fluorescent signals in the first generation of rescued viruses and demonstrated replication capabilities in plaque and hemagglutination assays. The recombinant viruses based on the NA and NS genes were capable of stably expressing Nluc across different generations, and exhibited correct fluorescent distribution patterns in mouse infection experiments. Meanwhile, the NS gene-based recombinant virus demonstrated superior stability in the mouse model.Conclusions:This study demonstrates that the NS gene segment of influenza virus can serve as an effective insertion site for exogenous genes without impairing the viral replication or infectivity, and the recombinant virus constructed based on it exhibits high integration stability and substantial application potential.

Objective:To investigate the impact of inserting an exogenous gene, NanoLuc (Nluc), at different sites in the influenza virus genome on viral properties and analyze the expression stability of the exogenous gene both in vitro and in vivo. Methods:Using molecular cloning techniques and reverse genetics, eight recombinant influenza viruses were constructed by inserting the exogenous Nluc gene into the gene segments encoding hemagglutinin (HA), neuraminidase (NA), non-structural protein (NS), and polymerase basic protein 1 (PB1). Viral replication capacity was evaluated by hemagglutination and plaque assays. Nluc expression in infected cells was monitored by fluorescence imaging. The potential impact of the exogenous gene insertion on viral infectivity was examined in a mouse infection model. Independent samples t-test were used for statistical analysis. Results:The recombinant viruses with insertions in the HA, NA, and NS gene segments generated fluorescent signals in the first generation of rescued viruses and demonstrated replication capabilities in plaque and hemagglutination assays. The recombinant viruses based on the NA and NS genes were capable of stably expressing Nluc across different generations, and exhibited correct fluorescent distribution patterns in mouse infection experiments. Meanwhile, the NS gene-based recombinant virus demonstrated superior stability in the mouse model.Conclusions:This study demonstrates that the NS gene segment of influenza virus can serve as an effective insertion site for exogenous genes without impairing the viral replication or infectivity, and the recombinant virus constructed based on it exhibits high integration stability and substantial application potential.

Objective:To quantitatively analyze the plaque components of carotid artery through energy spectrum computed tomography angiography(CTA),and to measure the blood flow perfusion in the blood-supply area of carotid artery through CT perfusion(CTP),so as to explore the relationship among plaque component,the degree of luminal stenosis and cerebral blood flow perfusion.Methods:A total of 68 patients with unilateral plaques of carotid artery and severe vascular stenosis who were screened and diagnosed by ultrasound and CTA in Xiyuan Hospital from December 2017 to July 2019 were selected,and all patients underwent CTA examination and CTP examination.North American symptomatic carotid endarterectomy test(NASCET)method was used to measure the degree of carotid stenosis.The GE AW 4.7 post-process workstation was used to conduct analyses of energy spectrum and cerebral perfusion for the plaque component.And then,the slope of energy spectrum curve and the effective atomic number were obtained.At the same time,the cerebral blood volume(CBV),cerebral blood flow(CBF),time to peak(TTP)and mean transit time(MTT)of contrast agent in blood-supplying area of anterior cerebral artery(ACA)and middle cerebral artery(MCA)at the side of lesion were measured.Results:A total of 68 measured plaques of 68 patients met the condition,including 44 vulnerable plaques(including lipid plaques and mixed plaques)and 24 stable plaques(fibrous plaques).The average slopes of the energy spectrum curves of vulnerable plaque and stable plaque were respectively 0.45±0.45 and 1.15±0.39,and the differences were significant(t=2.413,P<0.05).The averagely effective atomic numbers of vulnerable plaques and stable plaques were respectively 7.21±1.06 and 8.01±0.63,and the difference were significant(t=2.548,P<0.05).The average TTP values of the ACA at the side of lesion of vulnerable plaques and stable plaques were respectively(12.20±1.61)S and(13.59±2.79)S,and the difference was significant(t=-2.607,P<0.05).The mean MTT values of the ACA at the side of lesion of vulnerable plaques and stable plaques were respectively(5.07±1.66)S and(6.09±2.19)S,and the difference was significant(t=-2.177,P<0.05).The degree of vascular stenosis at the side of lesion was positively correlated with TTP and MTT in blood-supplying area of middle cerebral artery(MCA)at the side of lesion(r=0.537,0.465,P<0.05),and that was negatively correlated with CBF values in blood-supplying areas of ACA and MCA at the side of lesion(r=-0.281,-0.569,P<0.05),respectively.The slope of the energy spectrum curve of carotid plaque was positively correlated with the TTP values in blood-supplying areas of ACA and MCA at the side of lesion(r=0.242,0.246,P<0.05),respectively.Conclusion:CT spectral imaging can quantitatively analyze the displayed components of carotid atherosclerotic plaque,and the degree of vascular stenosis can affect the blood flow perfusion of cerebral tissue,and the delays of TTP and MTT are more easily caused by vulnerable plaque,and the TTP of them is more sensitivity.

Objective:To evaluate the performance of the influenza surveillance network and compare the epidemic intensity of influenza during 2014-2020 in Shanghai.Methods:Based on the weekly reports of influenza-like illness (ILI) and laboratory-confirmed influenza cases from January 1, 2014 to December 31, 2020. This study first evaluated the data reporting and specimen collection of ILI cases for each sentinel hospital, and then calculated the percentage of ILI (ILI%), the proportion of specimens tested positive for influenza, and the incidence of influenza among all ILI outpatient and emergency visits to measure the epidemic intensity of influenza. Finally, seasonal autoregressive integrated moving average (ARIMA) model was applied to quantify the changes in epidemic intensity of influenza in 2020.Results:The proportion of influenza surveillance sentinel hospitals with a score of less than 5 in the evaluation of ILI data reporting and samples collection were 9.68% and 21.05% in 2020 in Shanghai, respectively. ILI% was estimated to be 1.51% (95% CI: 1.50%-1.51%) and 2.31% (95% CI: 2.30%-2.32%), respectively for 2014-2019 and 2020; the proportion of specimens tested positive was 24.27% (95% CI: 24.02%- 24.51%) and 7.15% (95% CI: 6.78%-7.54%), respectively; and the incidence of influenza was 3.66‰ (95% CI: 3.62‰-3.70‰) and 1.65‰ (95% CI: 1.57‰-1.74‰), respectively. ARIMA model showed that ILI% was increased by 45.25% in 2020 in Shanghai, and the proportion of specimens tested positive and the incidence of influenza were reduced by 78.45% and 51.80%, respectively. Conclusions:In 2020, the performance of influenza surveillance system has changed, ILI% has increased, the proportion of specimens tested positive and the incidence of influenza has decreased in Shanghai. The change in the quality of influenza surveillance is also a potential factor affecting the epidemic intensity of influenza. In the future, the quality control of influenza surveillance network still needs to be further strengthened.

ObjectiveTo conduct on-site epidemiological investigation， emergency response， tracing of infection source and analysis of a confirmed COVID-19 case of a foreign airline cargo service staff member in Shanghai’s international airport， aiming to provide reference for prevention of imported COVID-19 cases under regular prevention and control of COVID-19. MethodsA retrospective field epidemiological investigation was conducted to collect information of basic characteristics， illness onset， diagnosis， treatment， clinical manifestations， exposure history and risk factors within 14 days before onset， close contacts， close contacts of close contacts， and key places related to activity trajectories. Respiratory tract specimens of cases and contacts were collected for detection of SARS-CoV-2 by real time RT-PCR （rRT-PCR）. Emergency response， including infection source analysis and contact management， was conducted. ResultsThe case developed pharyngeal itch on July 28， 2021， and fever on the 30th， and went to the hospital for treatment twice on the 31st. Because his specimen was positive for SARS-CoV-2 by rRT-PCR on August 1， he was isolated and treated on August 2 and diagnosed as a confirmed case of COVID-19. The case was a foreign airline cargo service member at an international airport. The two regular nucleic acid screenings of him as a high-risk occupation on July 21 and 28 were negative. He did not leave Shanghai within 14 days before the onset of illness. During July 22nd and 23rd， he was repeatedly exposed during work to the high-risk environment that may have been contaminated by SARS-CoV-2 and had contact with the crew of foreign airlines， and the personal protection was not standardized. None of the 67 close contacts and 567 close contacts of close contacts in Shanghai showed symptoms during the 14-day medical quarantine， and the specimens of them were all negative for SARS-CoV-2 by rRT-PCR. The results of genome sequencing analysis showed that the genomic homology between the virus of the case and the one of recent domestic local epidemic and the recent imported cases was low， and the homology with the overseas Delta mutant strain was higher than that of the domestic Delta mutant strain. ConclusionThe situation of prevention of COVID-19 import is still serious. It is necessary to conduct regular nucleic acid screening for high-risk occupational groups， strengthen the diagnosis and reporting awareness of medical institutions， effectively implement the prevention and control measures for people， objects， and environment at international airports， and further enhance the public's awareness of personal protection.

Objective:To evaluate narrow band imaging-magnifying endoscopy (NBI-ME) for the further assessment of lesions of low-grade intraepithelial neoplasia (LGIN) in the gastric biopsy.Methods:Data of 180 patients who underwent NBI-ME before endoscopic submucosal dissection (ESD) for biopsy of gastric LGIN at the First Affiliated Hospital of Soochow University from January 2017 to October 2020 were analyzed retrospectively. Taking the pathological results after ESD as the gold standard, the sensitivity, the specificity, the positive predictive value, the negative predictive value, and the accuracy of NBI-ME in predicting the pathological upgrading of gastric LGIN lesions after ESD were calculated, and the receiver operator characteristic (ROC) curve was drawn.Results:Among 180 gastric LGIN lesions, 115 (63.89%) were pathological upgraded and 65 (36.11%) were not after ESD. There were 10 missed diagnoses, 19 misdiagnoses, and 151 correct diagnoses in NBI-ME examination before ESD. The sensitivity, the specificity, the positive predictive value, the negative predictive value, and the accuracy of NBI-ME in predicting the pathological upgrading of gastric LGIN lesions after ESD were 91.3% (105/115), 70.8% (46/65), 84.7% (105/124), 82.1%(46/56) and 83.9% (151/180), respectively. The area under the ROC curve was 0.810 (95% CI: 0.737-0.883). Conclusion:Further NBI-ME examination of gastric LGIN lesions diagnosed by biopsy pathology can accurately predict whether the lesions have pathological upgrading after ESD, which is of important guiding significance for the patients to choose the treatment strategy of further follow-up or endoscopic resection.

Objective:To discuss the challenges and problems of the prevention and control of emerging infectious diseases in Shanghai as a megacities.Methods:An imported case of cutaneous anthrax occurred in Shanghai on May 30, 2019. Shanghai Municipal Center for Disease Control & Prevention conducted an epidemiological investigation and treatment of the cases; after this case, the data of patients with cutaneous anthrax were collected, and an epidemiological study was conducted. Meanwhile, the wound and blood samples of the patient were collected for laboratory testing.Results:Of the seven wound samples of the patient, 6 were positive for the Bacillus anthracis nucleic acid test, and the double serological test results showed a 4-fold increase in the titer of anthrax antibodies. Shanghai CDC conducted an epidemiological investigation of the confirmed cases and observed its contacts. After treatment, the patients recovered, and no other issues appeared among the 19 contacts. Conclusions:Shanghai must strengthen the training of clinicians on emerging infectious diseases to achieve early detection, diagnosis, and treatment of imported infectious diseases and reduce the incidence, spread, and death of the diseases. At the same time, multi-department joint prevention and control are needed to prevent and control secondary cases.

Objective To investigate the effect of aldehyde-free fixation solution and aldehyde fixation solution on DNA and cytoplasm of nucleus.Methods The DNA quantification (IOD),discrete coefficient (CV),DNA index (DI) and nucleus area (area) were measured by DNA quantification system (ICM) in 8 different fixation solution;combined with cytological staining,the effects of different fixative solution on nuclear DNA and cytoplasmic staining were analyzed.Results Aldehyde fixation solution was better than aldehyde free fixation solution,the formaldehyde in fixtion solution has important influence on Feulgen-eosin staining.Conclusion Aldehyde fixation solution combined with cytological staining can obtained good dyeing effect,which provides the basis for the subsequent multi-technique joint diagnosis.

Objective To study the aberrant DNA methylation of H19 and MEST in infertile male sperm.Methods Routine semen analysis and morphological analysis were performed on 15 control individuals and 33 oligoastheno-teratozoospermias.Sperms were purified by density gradient centrifugation and DNA was extracted and treated with bisulfite.After PCR,the purified PCR products were cloned into pMD (R)18-T vector and proceed to chemical transformation,restriction enzyme reaction.For each sample,positive clones were selected for sequencing analysis and DNA methylation analysis.Results Five control individuals had high degree of H19 and methylation rate was 100.0％,while the methylation rate of 93.0％ in 10 oligoastheno-teratozoospermias was significantly lower.Significant difference was found in methylation rate between the control individuals and oligoastheno-teratozoospermias (P ＜ 0.01).As to MEST,10 control individuals had a low degree of MEST and the methylation rate was 1.6％,while the methylation rate in 23 oligoastheno-teratozoospermias was 4.8％.No significant difference was found in the methylation rate between control individuals and oligoastheno-teratozoospermias (P ＞ 0.05).Conclusion Male infertility is associated with aberrant methylation of H19 differentially methylated region,and it may be a biomarker of human spermatogenesis defect.