Objective To develop and verify a multiplex fluorescent quantitative PCR method for detection of common bacterial and fungal contaminants in cell culture medium.Methods According to NUC gene of Staphylococcus aureus,COLA gene of Clostridium spore,ITS-2 segment sequence of Candida albicans,a set of primers and probes were designed for each respectively,and using UBI3 gene of capsicum introduced as external standard gene,a triple reaction system of Staphylococcus aureus,Clostridium spore and external standard gene and a double reaction system of Candida albicans and external standard gene were established.The primer specificity,linear range,limit of detection,specificity,anti-interference performance and precision of the method were verified.Finally,100 samples of 293T cell culture medium were detected by using the developed method,which was compared with the common PCR method.Results Three pairs of primers all amplified about 100 bp specific gene bands corresponding to the three strains at different annealing temperatures(56,57,58 and59 ℃),and the size was consistent with the expected.In the range of 5.80 × 10~6 — 5.80 × 10~2 copies/μL,the standard plasmids of the three strains showed a good linear relationship with the Ct values.The standard curve equations were:Y=-3.373 X+37.48,Y=-3.557X+36.59 and Y=-3.536 X+39.78,each R~2> 0.99,respectively,and the amplification efficiency was in the range of 90%—110%.All the limits of detection of the three strains were 10~1 CFU/mL.The primers and probes of the three strains showed no specific amplification on the genomic DNA of six kinds of cells that were prone to cross-reaction.The genomic DNA of 293T cells,Yeast,Escherichia coli and Mycoplasma sp.had no effect on the detection.The CVs of repeatability and intermediate precision verification were both less than 15%.Among 100 cell culture medium samples,14 positive and 86 negative samples were detected,and the results of common PCR method for three positive and two negative samples randomly selected were consistent with the developed method.Conclusion The multiplex fluorescent quantitative PCR method developed in this study for the detection of bacteria and fungi in cell culture medium has good specificity,anti-interference performance and precision,and is simple to operate with low cost and high sensitivity,which can quickly detect the contaminants during cell culture.

Objective : To understand the epidemiological characteristics of notifiable respiratory infectious diseases in Huzhou City, Zhejiang Province from 2017 to 2022, so as to provide insights into formulation of respiratory infectious disease prevention and control strategies. Methods: The data pertaining to notifiable respiratory infectious disease in Huzhou City from 2017 to 2022 were collected through the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. Epidemiological characteristics of notifiable respiratory infectious disease was analyzed using a descriptive epidemiological method. : Results Conclusions: A total of 31 314 cases of notifiable respiratory infectious diseases were reported in Huzhou City from 2017 to 2022, with an average annual reported incidence of 169.12/105. The reported incidence of notifiable respiratory infectious diseases appeared a tendency towards a rise in Huzhou City from 2017 to 2022 (P<0.05). The top six reported diseases in terms of case numbers were influenza (20 048 cases), tuberculosis (6 920 cases), COVID-19 (1 893 cases), mumps (1 413 cases), pertussis (475 cases) and scarlet fever (442 cases), accounting for 99.61% of the total cases. The incidence of influenza, COVID-19 and pertussis showed a tendency towards a rise, the incidence of mumps and tuberculosis showed a tendency towards a decline (all P<0.05), and scarlet fever remained at a low-level incidence (P>0.05). Respiratory infectious diseases were mainly reported in winter (January, February and December), with 14 644 cases accounting for 46.77%. There were 15 068 cases reported in schools and kindergartens, accounting for 48.12%. The incidence showed a U-shaped variation with age, with the highest incidence in residents at ages of 10 years and below (987.68/105), and showing a tendency towards a rise in residents at ages of 60 years and above. The incidence of respiratory infectious diseases in Huzhou City from 2017 to 2022 increased significantly. Influenza, tuberculosis, COVID-19, mumps and pertussis are key notifiable respiratory infectious diseases. Residents at ages of 10 years and below and 60 years and above should be given a high priority for respiratory infectious disease control.

Objective To develop and verify a multiplex fluorescent quantitative PCR method for detection of common bacterial and fungal contaminants in cell culture medium.Methods According to NUC gene of Staphylococcus aureus,COLA gene of Clostridium spore,ITS-2 segment sequence of Candida albicans,a set of primers and probes were designed for each respectively,and using UBI3 gene of capsicum introduced as external standard gene,a triple reaction system of Staphylococcus aureus,Clostridium spore and external standard gene and a double reaction system of Candida albicans and external standard gene were established.The primer specificity,linear range,limit of detection,specificity,anti-interference performance and precision of the method were verified.Finally,100 samples of 293T cell culture medium were detected by using the developed method,which was compared with the common PCR method.Results Three pairs of primers all amplified about 100 bp specific gene bands corresponding to the three strains at different annealing temperatures(56,57,58 and59 ℃),and the size was consistent with the expected.In the range of 5.80 × 10~6 — 5.80 × 10~2 copies/μL,the standard plasmids of the three strains showed a good linear relationship with the Ct values.The standard curve equations were:Y=-3.373 X+37.48,Y=-3.557X+36.59 and Y=-3.536 X+39.78,each R~2> 0.99,respectively,and the amplification efficiency was in the range of 90%—110%.All the limits of detection of the three strains were 10~1 CFU/mL.The primers and probes of the three strains showed no specific amplification on the genomic DNA of six kinds of cells that were prone to cross-reaction.The genomic DNA of 293T cells,Yeast,Escherichia coli and Mycoplasma sp.had no effect on the detection.The CVs of repeatability and intermediate precision verification were both less than 15%.Among 100 cell culture medium samples,14 positive and 86 negative samples were detected,and the results of common PCR method for three positive and two negative samples randomly selected were consistent with the developed method.Conclusion The multiplex fluorescent quantitative PCR method developed in this study for the detection of bacteria and fungi in cell culture medium has good specificity,anti-interference performance and precision,and is simple to operate with low cost and high sensitivity,which can quickly detect the contaminants during cell culture.

Objective: To investigate the epidemilogical characteristics of pulmonary tuberculosis in Huzhou City, Zhejiang Province from 2014 to 2023, so as to provide the basis for formulating prevention and control measures for the construction of "TB-free city". Methods: The data of pulmonary tuberculosis cases in Huzhou City from 2014 to 2023 was collected through the Infectious Disease Reporting Management System of Chinese Disease Prevention and Control Information System. The onset time, region, and population distribution characteristics of the cases were described. Results: A total of 11 598 cases of pulmonary tuberculosis were reported in Huzhou City from 2014 to 2023, with an average annual incidence of 37.42/105. The reported incidence decreased from 47.50/105 in 2014 to 28.36/105 in 2023 (P<0.05), with an annual decline rate of 5.57%. There were 6 304 etiological positive cases, accounting for 54.35%. The peak season for pulmonary tuberculosis cases was from March to September, with the highest seasonal ratio of 112.48% in May. The average annual reported incidence rates in Anji County and Changxing County were relatively high (46.14/105 and 41.15/105). The reported incidence rate of pulmonary tuberculosis in Huzhou City increased with age (P<0.05), peaking at 97.36/105 in the group aged 75 to <80 years. There were 7 991 male pulmonary tuberculosis cases and 3 607 female cases, with a male-to-female ratio of 2.22∶1. The average annual incidence rates of pulmonary tuberculosis was higher in males than in females (50.39/105 vs. 23.87/105). Farmers were the primary occupation affected, with 6 350 cases accounting for 54.75%. Conclusions The reported incidence rate of pulmonary tuberculosis in Huzhou City decreased from 2014 to 2023, with a high incidence in spring and summer. The incidence rates in Anji County and Changxing County were higher than Huzhou City's average. Male, elderly residents and farmers were the key populations for pulmonary tuberculosis prevention and control.

Objective: To investigate the trends in incidence and epidemiological characteristics of notifiable infectious diseases in Huzhou City, Zhejiang Province from 2005 to 2023, so as to provide insights into optimizing infectious disease surveillance and control. Methods: Data of notifiable infectious disease cases in Huzhou City from 2005 to 2023 were collected from the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. Trends in incidence were analyzed using annual percent change (APC) and average annual percent change (AAPC). The population distribution and seasonal characteristics were descriptively analyzed. Results: From 2005 to 2023, a total of 504 283 cases of notifiable infectious diseases were reported in Huzhou City, with an annual crude incidence rate of 892.65/105 and a standardized incidence rate of 989.21/105. The incidence rate of notifiable infectious diseases in Huzhou City showed an upward trend (AAPC=8.886%, P<0.05), of which there was an obvious upward trend from 2021 to 2023 (APC=95.996%, P<0.05). After the removal of COVID-19 incidence, the incidence trend was basically unchanged (AAPC=7.970%, P<0.05). From 2005 to 2023, the incidence rate of class A and B notifiable infectious diseases showed no obvious trend (P>0.05), and the incidence rate of class A and B respiratory infectious diseases showed an upward trend (AAPC=6.958%, P<0.05). After the removal of COVID-19 incidence, the two showed a downward trend (AAPC=-7.680% and -8.660%, both P<0.05). The incidence rate of class A and class B intestinal infectious diseases, blood-borne and sexually transmitted infectious diseases showed a downward trend (AAPC=-14.849% and -5.977%, both P<0.05), while the incidence rates of natural and insect-borne infectious diseases did not show a significant trend (P>0.05). The overall incidence rate of class C infectious diseases showed an upward trend (AAPC=13.058%, P<0.05). The incidence rate ratio (IRR) of notifiable infectious diseases between males and females was 1.26 (95%CI: 1.25-1.27). A total of 204 043 cases under 10 years were reported, accounting for 40.46%. The peak incidence of class A and B respiratory infectious diseases was in January, May and June, while that of class A and B intestinal infectious diseases was from June to October. The peak incidence of class C respiratory infectious diseases was in January, March, April and December, while that of class C intestinal infectious diseases was from May to August and from November to December. Conclusions The incidence rate of notifiable infectious diseases in Huzhou City showed an upward trend from 2005 to 2023, which was more obvious from 2021 to 2023. Men and children under 10 years were the high-risk population. The incidence of respiratory and intestinal infectious diseases had obvious seasonal characteristics.

Objective: To analyze the epidemiological characteristics of pertussis in Huzhou City, Zhejiang Province from 2012 to 2022, so as to provide insights into formulation of pertussis control measures. Methods: The data of reported pertussis cases in Huzhou City from 2012 to 2022 were collected through the Infectious Disease Report Information System of Chinese Disease Prevention and Control Information System. The temporal, spatial and population distributions of pertussis cases in Huzhou City from 2012 to 2022 were analyzed using a descriptive epidemiological method. Results: A total of 499 pertussis patients were reported in Huzhou City from 2012 to 2022, with mean annual reported incidence of 1.508/105, and no death was reported. The reported incidence of pertussis remained at a low level in Huzhou City from 2012 to 2021, all below 1/105, and increased to 12.625/105 in 2022. The reported incidence of pertussis appeared an overall tendency towards a rise in Huzhou City from 2012 to 2021 (Z=-29.261, P<0.001). The incidence of pertussis peaked from June to July, and a relatively higher incidence rate of pertussis was reported in Deqing (6.359/105) and Anji counties (1.725/105), while higher incidence was found among children at ages of <1 year (30.566/105), 4 years (31.896/105) and 5 years (29.485/105). Conclusion The reported incidence of pertussis was at a low level in Huzhou City from 2012 to 2021, and increased sharply in 2022. The incidence of pertussis peaked from June to July, was concentrated in Deqing and Anji counties and higher among infants under one year of age and preschool children at ages of 4 to 5 years.

Objective To investigate the resistance level of adult Aedes albopictus to commonly used insecticides in urban areas of Huzhou City， and analyze the trend of insecticide resistance. Methods Aedes albopictus larvae were collected from different urban areas of Huzhou City in 2019 and 2020. The larvae were reared to adults for determining resistance using the WHO standard test kits by contact tube method. Results The mortality was 97.06%‒100.00% in adult Aedes albopictus mosquitoes exposed to 0.4% beta-cypermethrin， 0.1% deltamethrin， 0.5% malathion， 0.05% propoxur and 3% permethrin in 2019. While the adult mosquitoes exposed to 0.4% beta Cypermethrin， 0.08% beta Cypermethrin， 0.03% deltamethrin， 0.4% permethrin， 0.5% malathion and 0.02% propoxur in 2020， the adjusted mortality were 98.58%， 89.09%， 85.64%， 91.34%， 91.43% and 100% respectively . When the insecticide doses on medicated paper were high， the Huzhou populations of Aedes albopictus were sensitive to beta-cypermethrin， deltamethrin， malathion and propoxur， and showed suspected resistance to permethrin. While the Aedes albopictus were exposed to low testing dose， they showed suspected resistance to beta-cypermethrin， deltamethrin， permethrin and malathion， but remained to be sensitive to propoxur. Conclusion The adult Aedes albopictus in Huzhou City from 2019 to 2020 was still sensitive to propoxur， but their resistance to malathion changes from sensitive to resistant. When the contact tube method is used for insecticide resistance， it is suggested that the accurate diagnostic dose should be obtained first before conducting further insecticide resistance tests.

Objective:To explore the effect of mild cognitive impairment on the subjective well-being of the elderly and the mediating role of self-efficacy in it. To provide an effective basis for health management and clinical intervention in the elderly with mild cognitive impairment.Methods:A total of 164 patients with mild cognitive impairment from December 2018 to June 2019 were surveyed by the General Self-efficacy Scale, the Montreal Cognitive Assessment Scale, and the Memorial University of Newfoundland Scale of Happiness Scale.Results:The overall score of self-efficacy was 21.41±3.98, and subjective well-being was 26.44±7.35. There was a positive correlation among cognitive function, self-efficacy and subjective well-being, except that there was no significant correlation between self-efficacy and cognitive function( r values were 0.263-0.687, P<0.01). Self-efficacy played a part in mediating between cognitive function and subjective well-being in patients with mild cognitive impairment, accounting for 37.8% of the total effect. Conclusions:Mild cognitive impairment can also have a negative impact on the subjective well-being of the elderly. Among them, self-efficacy plays a particularly important role in mediating. Interventions about self-efficacy on the elderly with mild cognitive impairment may have more significant effects.

Objective:To explore the current status and hot trends of geriatric nursing in China in the past five years, in order to provide a reference for related research.Methods:This research adopted the bibliometric research method. The Wanfang database, China National Knowledge Infrastructure (CNKI) , and China Biomedical Literature Database were searched on October 30, 2020, and bibliometric analysis was carried out based on the literature related to geriatric nursing published from 2016 to 2020.Results:A total of 4 930 papers included were published in 359 journals. The cumulative number of papers published in the top 15 journals exceeded 50%, and one third of them were non-nursing journals. The number of papers published in the region where the first author of the paper was located was from 1 to 555, and the top 3 regions were Beijing, Jiangsu, and Guangdong respectively. Research hotspots were mainly concentrated in five areas, namely, common clinical problem nursing in elderly patients, geriatric nursing services in institutions and communities, quality of life and psychological nursing, chronic disease management, and safety risk management.Conclusions:In the past five years, geriatric nursing research has been large in volume and published in journals. The research attention of different cities is quite different, and the research field is continuously expanding, which is in line with the background of my country's population aging and the guiding direction of national policies.

ObjectiveTo investigate the features and changing trend of drug-induced liver injury (DILI) in the elderly from 2009 to 2019, and to provide a reference for clinical prevention and treatment of DILI in the elderly. MethodsA retrospective analysis was performed for the clinical data of 2107 elderly patients, aged ≥60 years, who were diagnosed with DILI in The Fifth Medical Center of Chinese PLA General Hospital from January 2009 to December 2019, and they were divided into groups according to age. Related clinical data were analyzed, including age, sex, clinical features, prognosis, and regional distribution. The Chi-square test was used for comparison of categorical data between groups. ResultsAmong the 2107 patients with DILI, there were 802 male patients and 1305 female patients, with a male/female ratio of 1∶1.63. Cholestasis type was the most common clinical type and was observed in 1439 patients (68.3%). There was the highest number of patients in the 60-64 years group (942 patients, 44.7%), among whom 618(65.6%) were female, 589(62.5%) had cholestasis type, 471(50.0%) had chronic DILI, 421(44.7%) had drug-induced liver cirrhosis, and 25(2.7%) had drug-induced liver failure. There were 187 patients in the 75-79 years group, among whom 110 (58.8%) patients were male, 137(73.3%) had cholestasis type, 114(60.9%) had liver cirrhosis, 4(2.1%) had drug-induced liver failure. The results showed that chronic DILI was more common in the 60-64 years group, and liver cirrhosis was more common in the 75-79 years group. As for prognosis, in the 60-64 years group, 27 patients (2.9%) were cured, 885 (93.9%) were improved, 30(32%) had no response or died; in the 65-69 years group, 16 (2.8%) were cured, 528 (92.0%) were improved, and 30(5.2%) had no response or died; in the 70-74 years group, 9(2.8%) were cured, 305(94.1%) were improved, and 10 (3.6%) had no response or died. The results showed that there was no significant difference in mortality rate between the different age groups (P＞0.05). The proportion of elderly DILI patients among hospitalized DILI patients increased from 15.90% in 2009 to 22.05% in 2013 and 27.51% in 2019, with a 1.73-fold increase in 11 years. As for regional distribution, the patients in North China accounted for the highest proportion of 47.08% (the patients from Hebei, Shanxi, and Inner Mongolia accounted for 24.92%, 10.96%, and 10.25%, respectively), followed by those in Northeast China who accounted for 17.85%. The patients in Beijing accounted for 11.53%. ConclusionThe proportion of elderly DILI patients among hospitalized DILI patients tends to increase in these years. Cholestasis type is the most common clinical type, and most of the patients with this clinical type progress to chronic DILI and drug-induced liver cirrhosis. Early diagnosis, early intervention, and standardized treatment of elderly DILI should be taken seriously.