Objective: To examine the impact of temperature on the incidence of hand-foot-mouth disease (HFMD), so as to provide a basis for research on the impact of climate on HFMD. Methods: HFMD incidence data in Huzhou City, Zhejiang Province from 2010 to 2022 were collected from the Infectious Disease Surveillance System of Chinese Disease Prevention and Control Information System. Concurrent meteorological data were obtained from the Huzhou Meteorological Bureau. A distributed lag non-linear model (DLNM) was employed to analyze the impact of temperature on HFMD incidence. Results: A total of 65 059 cases of HFMD were reported in Huzhou City from 2010 to 2022, with incidence peaks occurring from April to July and from October to November. The 5th, 25th, 50th, 75th, and 95th percentiles (P5, P25, P50, P75, and P95) of daily average temperature were 2.5, 9.4, 17.8, 24.7, and 30.6 ℃, respectively. The results from the DLNM analysis revealed a non-linear relationship between daily average temperature and HFMD incidence, which exhibited an irregular "M" shape. The incidence peaks were observed at 6.5 ℃ (RR=1.596, 95%CI: 1.154-2.207) and 25.7 ℃ (RR=3.257, 95%CI: 2.614-4.058), respectively. The maximum single-day effect was observed at a daily average temperature of -6.2 ℃ at lag 0 day (RR=1.250, 95%CI: 1.100-1.420), while the minimum single-day effect was observed at a daily average temperature of 34.6 ℃ at lag 0 day (RR=0.881, 95%CI: 0.826-0.940). The impact of daily average temperature at P5 on the incidence of HFMD showed RR values greater than 1 at lag 0, 3, 27, and 30 days, but less than 1 at lag 9 and 12 days, with no significant cumulative lag effect. In contrast, for the daily average temperature at P95, the RR values less than 1 at lag 15, 18, and 21 days, with significant cumulative lag effects identified. Conclusions The incidence of HFMD in Huzhou City peaks from April to July and again from October to November. There exists a nonlinear relationship and lag effect between temperature and HFMD incidence, with particular attention drawn to the impact of extreme temperatures on the disease's onset.

Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Diabetic cardiomyopathy (DCM) is a medical condition characterized by cardiac remodeling and dysfunction in individuals with diabetes mellitus. Sarcoplasmic reticulum (SR) and mitochondrial Ca²⁺ overload in cardiomyocytes have been recognized as biological hallmarks in DCM; however, the specific factors underlying these abnormalities remain largely unknown. In this study, we aimed to investigate the role of a cardiac-specific long noncoding RNA, D830005E20Rik (Trdn-as), in DCM. Our results revealed the remarkably upregulation of Trdn-as in the hearts of the DCM mice and cardiomyocytes treated with high glucose (HG). Knocking down Trdn-as in cardiac tissues significantly improved cardiac dysfunction and remodeling in the DCM mice. Conversely, Trdn-as overexpression resulted in cardiac damage resembling that observed in the DCM mice. At the cellular level, Trdn-as induced Ca²⁺ overload in the SR and mitochondria, leading to mitochondrial dysfunction. RNA-seq and bioinformatics analyses identified calsequestrin 2 (Casq2), a primary calcium-binding protein in the junctional SR, as a potential target of Trdn-as. Further investigations revealed that Trdn-as facilitated the recruitment of METTL14 to the Casq2 mRNA, thereby enhancing the m⁶A modification of Casq2. This modification increased the stability of Casq2 mRNA and subsequently led to increased protein expression. When Casq2 was knocked down, the promoting effects of Trdn-as on Ca²⁺ overload and mitochondrial damage were mitigated. These findings provide valuable insights into the pathogenesis of DCM and suggest Trdn-as as a potential therapeutic target for this condition.
Animals ; Diabetic Cardiomyopathies/pathology* ; RNA, Long Noncoding/genetics* ; Myocytes, Cardiac/metabolism* ; Mice ; Calsequestrin/genetics* ; Calcium/metabolism* ; Male ; Sarcoplasmic Reticulum/metabolism* ; Methyltransferases/metabolism* ; Mice, Inbred C57BL ; Mitochondria, Heart/metabolism* ; Disease Models, Animal ; Mitochondria/metabolism*

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 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 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 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.