Objective To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. Methods B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Results Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ² = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. Conclusion The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system.

Objective To examine the association between epidemiological characteristics of hemorrhagic fever with renal syndrome (HFRS) and meteorological factors in Shenzhen City during the period from 2012 to 2019. Methods Average atmospheric pressure, average air temperature, average relative humidity, precipitation, wind speed, and sunshine duration were captured from Meteorological Bureau of Shenzhen City each month from 2012 to 2019. The average monthly rodent densities in Shenzhen City from 2012 to 2019 were acquired from the Vector Surveillance Management System of Guangdong Provincial Center for Disease Control and Prevention, and the monthly HFRS incidence was retrieved from Shenzhen Municipal Disease Surveillance System from 2012 to 2019. The correlation between meteorological factors and the monthly incidence of HFRS was examined us ing Spearman’s rank correlation in Shenzhen City, and the temporal trends in monthly HFRS incidence and the degrees of freedom for the rodent density were determined in Shenzhen City with a generalized additive model. The optimal lag time was identified using excess risk (ER) and its 95% confidence interval (CI), and univariate and multivariate models were fitted to evaluate the impact of meteorological factors on HFRS incidence in Shenzhen City. Results The median number of incident HFRS cases was 3.00 (interquartile range, 3.25) in Shenzhen City from 2012 to 2019, with an average air temperature of (23.44 ± 4.91) °C, average relative humidity of (76.05 ± 7.61)%, median precipitation of 4.10 (interquartile range, 6.83) mm, average wind speed of (1.97 ± 0.26) m/s, average sunshine duration of (5.17 ± 1.64) h, and median monthly rodent density of 1.74% (interquartile range, 2.52%). Spearman’s rank correlation analysis showed that the average air temperature positively correlated with average relative humidity (r_s = 0.420, P < 0.05), precipitation (r_s = 0.658, P < 0.05) and sunshine duration (r_s = 0.633, P < 0.05), and the atmospheric pressure negatively correlated with average air temperature (r_s = −0.925, P < 0.05), relative humidity (r_s = −0.614, P < 0.05), precipitation (r_s = −0.789, P < 0.05) and sunshine duration (r_s = −0.437, P < 0.05), while the average relative humidity correlated positively with precipitation (r_s = 0.724, P < 0.05) and negatively with sunshine duration (r_s = −0.218, P < 0.05). Univariate modeling analysis showed that the ERs and their 95% CI were 0.639% (0.540%, 0.737%) for average atmospheric pressure, −7.157% (−8.113%, −6.190%) for average air temperature, −3.603% (−4.219%, −2.985%) for average relative humidity, −5.889% (−7.085%, −4.669%) forprecipitation,21.881% (−5.149%, 56.612%) for average wind speed, and −13.877% (−16.641%, −11.022%) for sunshine duation (all P values < 0.05). Multivariate modeling analysis showed that in the ensemble model combining average atmospheric pressure and precipitation, the highest ER (6.686%) was caused by increased average atmospheric pressure, and the highest absolute ER values for average air temperature (6.615%), average relative humidity (3.107%) and precipitation (5.386%) were seen after adjustment only for sunshine duration (all P values < 0.05), while the highest absolute ER for sunshine duration (11.875%) was found after adjustment for precipitation (P < 0.05). Conclusions An increase in average air temperature, relative humidity, precipitation and sunshine duration resulted in a reduced incidence rate of HFRS in Shenzhen City from 2012 to 2019, and an increase in average atmospheric pressure increased the incidence of HFRS. Meteorological factors are important determinants affecting HFRS incidence in Shenzhen City.

AIM:To investigate the modulatory role of E3 ubiquitin-protein ligase ITCH in Alzheimer disease(AD)-like pathology through the thioredoxin-interacting protein(TXNIP)-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway using both in vivo and in vitro experimental models.METHODS:(1)Ten 5×FAD(AD model)mice and 10 wild-type(WT)mice at 2-,4-and 6-month-old were randomly allocated into AD and WT groups.Amyloid β-protein(Aβ)plaque burden in the brain was detected by thioflavin-S and immunofluorescence staining,with the latter method additionally applied to assess TXNIP protein expression.The protein levels of ITCH and TXNIP were determined by Western blot,while their interaction was verified by co-immunoprecipitation.(2)Mouse mi-croglia BV2 cells stimulated by lipopolysaccharide(LPS)were used to construct neuroinflammation model,and were di-vided into control(CON)group and LPS+ATP treatment group.The BV2 cells stimulated by Aβ were used to construct AD inflammation model.According to the different treatment time,they were divided into CON,and 12,24 and 48 h treatment groups.Western blot was used to evaluate the expression of ITCH,TXNIP,and NLRP3 inflammasome compo-nents(NLRP3 and caspase-1)as well as the downstream IL-1β.Adenovirus-mediated ITCH overexpression(OE-ITCH)in Aβ-stimulated BV2 cells comprised three experimental groups:negative control group,Aβ oligomer stimulation group,and OE-ITCH group,with subsequent immunoblotting of inflammatory mediators.RESULTS:The deposition of Aβ plaques in the cortex and hippocampus of 5×FAD transgenic mice exhibited an age-dependent progression(P<0.01).Compared with WT mice,the levels of TXNIP protein increased synchronously,and the levels of ubiquitin ligase ITCH was significantly down-regulated(P<0.05).Co-immunoprecipitation confirmed the interaction between ITCH and TXNIP proteins in the brain of 2-and 4-month-old 5×FAD mice,which exhibited marked attenuation by 4 months of age.In BV2 microglial models,Aβ/LPS stimulation provoked significant ITCH suppression,concurrently up-regulating TXNIP,core NLRP3 inflammasome components(NLRP3 and caspase-1),and downstream IL-1β(P<0.05).Overexpression of ITCH significantly inhibited Aβ-induced activation of TXNIP and NLRP3 and therelated inflammatory factors in BV2 cells.CONCLUSION:The results of in vitro and in vivo experiments showed that ITCH protein exerts effects against AD-like pathology by inhibiting the expression of TXNIP-NLRP3 signaling pathway.

Objective:To analyze the epidemiological characteristics and trend of acute hemorrhagic conjunctivitis (AHC) in China from 2015 to 2022, and to provide evidence for the adjustment of prevention and control strategies and measures for AHC.Methods:The case data of AHC reported by national notifiable disease information system from 2015 to 2022 were collected, and descriptive analysis method were used to analyze the population distribution characteristics, temporal epidemiological trends and spatial clusters of AHC in China.Results:From 2015 to 2022, the incidence of AHC in China ranged from 1.85/100 000 to 2.97/100 000, with a fluctuating downward trend. The average annual percent change (AAPC) was -4.91 (95% CI: -7.74 to -2.00, P<0.05), with an annual percent change (APC) of 2.73 (95% CI: -2.34 to 8.06, P=0.189) for 2015—2019 and an APC of -14.23 (95% CI: -21.78 to -5.94, P<0.05). The age-specific incidence rate was highest in children aged 0-4 years (fluctuating between 4.69 and 5.67/100 000 from 2015 to 2019; It decreased significantly during 2020—2022, fluctuating between 1.93 and 2.72 per 100 000).The proportion of cases in children at 0-4 years of age showed a fluctuating downward trend from 8.68% in 2015 to 3.76% in 2020, with an increase in 2021—2022 to 5.74%. After 2020, the proportion of the population aged 60 years and above has increased, reaching 33.59% in 2022. Cases were mainly farmers, with a fluctuating upward trend of around 50% per year, with the highest percentage of 60.96% in 2020. The peak seasonal incidence of AHC was obvious from May to September from 2015 to 2019, but it was not obvious in 2020—2022. The cases were mainly distributed in Guangxi, Hainan and other provinces in the southwestern part of China. The high incidence counties were concentrated in Leye County of Guangxi, Maojian District of Hubei, Fuchuan Yao Autonomous County of Guangxi, Funing County of Yunnan, and Pulan County of Tibet every year. Conclusions:The overall epidemic rate of AHC in China showed a fluctuating downward trend from 2015 to 2022, with a pronounced decline observed between 2020 and 2022, potentially linked to non-pharmaceutical interventions during the COVID-19 pandemic. Increased attention needs to be given to farmers and people above 60 years of age to reduce the risk of morbidity. Moreover, prevention and control efforts should be strengthened in high-risk areas of southwestern China, and comprehensive measures should be implemented in counties with high incidence, including enhanced health education campaigns and improved allocation of sanitary facilities, to reduce the risk of AHC infection. This study is the first to highlight the potential impact of public health policies on AHC epidemiology, thereby offering a scientific foundation for population- and region-specific precision prevention strategies, particularly guiding the refinement of control measures in high-burden areas.

Thermal ablation (TA) is a widely applied minimally invasive treatment for benign thyroid nodules and low-risk papillary thyroid microcarcinoma. Compared to conventional surgery, TA offers advantages such as minimal trauma, rapid recovery, and no scarring. However, this procedure may lead to various complications, including intraoperative pain, nerve injury, hemorrhage, tracheal injury, skin burns, vasovagal reactions, nodule rupture, and thyroid dysfunction. Although TA demonstrates excellent safety and efficacy, further standardization of procedural protocols is necessary to minimize the incidence of complications.

Objective:To evaluate the predictive role of ultrasound, genetic testing, and clinical features in the malignancy risk of Bethesda Ⅲ thyroid nodules, and to explore strategies for optimizing treatment decisions.Methods:This retrospective study included 227 Bethesda Ⅲ thyroid nodules from patients who underwent surgical treatment at the Thyroid Surgery Department of Nanjing Drum Tower Hospital between Jan. 2020 and Dec. 2023. All patients underwent ultrasound evaluation and fine-needle aspiration. For nodules diagnosed as ultrasound, genetic testing, and clinical features were analyzed using univariate and multivariate regression to assess their association with malignancy.Results:Among the 227 nodules, 214 were malignant, resulting in a malignancy rate of 94.2%. The malignancy rate of thyroid nodules was 94.2%. In univariate analysis, age ( P=0.016), BRAF V600E gene mutation ( P<0.001), nodule size ( P=0.002), and TIRADS ( P<0.001) were significantly associated with malignancy in Bethesda Ⅲ thyroid nodules. Multivariate analysis confirmed that age ( OR=0.939, P=0.049) and BRAF V600E gene mutation ( OR=24.641, P<0.001) were significantly associated with thyroid nodule nature and served as independent predictive factors for malignancy. Conclusions:Genetic testing is an important method for predicting the malignancy of Bethesda Ⅲ thyroid nodules, and ultrasound also has high clinical value in assessing the malignancy risk of nodules. While some clinical features are highly correlated with nodule characteristics, they may not be practical in clinical application. For nodules classified as TIRADS 3 through ultrasound evaluation and negative for BRAF mutations, continued observation may be considered, whereas TIRADS 5 nodules or nodules with BRAF mutations should be prioritized for surgical treatment.

Thermal ablation (TA) is a widely applied minimally invasive treatment for benign thyroid nodules and low-risk papillary thyroid microcarcinoma. Compared to conventional surgery, TA offers advantages such as minimal trauma, rapid recovery, and no scarring. However, this procedure may lead to various complications, including intraoperative pain, nerve injury, hemorrhage, tracheal injury, skin burns, vasovagal reactions, nodule rupture, and thyroid dysfunction. Although TA demonstrates excellent safety and efficacy, further standardization of procedural protocols is necessary to minimize the incidence of complications.

Objective:To evaluate the predictive role of ultrasound, genetic testing, and clinical features in the malignancy risk of Bethesda Ⅲ thyroid nodules, and to explore strategies for optimizing treatment decisions.Methods:This retrospective study included 227 Bethesda Ⅲ thyroid nodules from patients who underwent surgical treatment at the Thyroid Surgery Department of Nanjing Drum Tower Hospital between Jan. 2020 and Dec. 2023. All patients underwent ultrasound evaluation and fine-needle aspiration. For nodules diagnosed as ultrasound, genetic testing, and clinical features were analyzed using univariate and multivariate regression to assess their association with malignancy.Results:Among the 227 nodules, 214 were malignant, resulting in a malignancy rate of 94.2%. The malignancy rate of thyroid nodules was 94.2%. In univariate analysis, age ( P=0.016), BRAF V600E gene mutation ( P<0.001), nodule size ( P=0.002), and TIRADS ( P<0.001) were significantly associated with malignancy in Bethesda Ⅲ thyroid nodules. Multivariate analysis confirmed that age ( OR=0.939, P=0.049) and BRAF V600E gene mutation ( OR=24.641, P<0.001) were significantly associated with thyroid nodule nature and served as independent predictive factors for malignancy. Conclusions:Genetic testing is an important method for predicting the malignancy of Bethesda Ⅲ thyroid nodules, and ultrasound also has high clinical value in assessing the malignancy risk of nodules. While some clinical features are highly correlated with nodule characteristics, they may not be practical in clinical application. For nodules classified as TIRADS 3 through ultrasound evaluation and negative for BRAF mutations, continued observation may be considered, whereas TIRADS 5 nodules or nodules with BRAF mutations should be prioritized for surgical treatment.

AIM:To investigate the modulatory role of E3 ubiquitin-protein ligase ITCH in Alzheimer disease(AD)-like pathology through the thioredoxin-interacting protein(TXNIP)-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway using both in vivo and in vitro experimental models.METHODS:(1)Ten 5×FAD(AD model)mice and 10 wild-type(WT)mice at 2-,4-and 6-month-old were randomly allocated into AD and WT groups.Amyloid β-protein(Aβ)plaque burden in the brain was detected by thioflavin-S and immunofluorescence staining,with the latter method additionally applied to assess TXNIP protein expression.The protein levels of ITCH and TXNIP were determined by Western blot,while their interaction was verified by co-immunoprecipitation.(2)Mouse mi-croglia BV2 cells stimulated by lipopolysaccharide(LPS)were used to construct neuroinflammation model,and were di-vided into control(CON)group and LPS+ATP treatment group.The BV2 cells stimulated by Aβ were used to construct AD inflammation model.According to the different treatment time,they were divided into CON,and 12,24 and 48 h treatment groups.Western blot was used to evaluate the expression of ITCH,TXNIP,and NLRP3 inflammasome compo-nents(NLRP3 and caspase-1)as well as the downstream IL-1β.Adenovirus-mediated ITCH overexpression(OE-ITCH)in Aβ-stimulated BV2 cells comprised three experimental groups:negative control group,Aβ oligomer stimulation group,and OE-ITCH group,with subsequent immunoblotting of inflammatory mediators.RESULTS:The deposition of Aβ plaques in the cortex and hippocampus of 5×FAD transgenic mice exhibited an age-dependent progression(P<0.01).Compared with WT mice,the levels of TXNIP protein increased synchronously,and the levels of ubiquitin ligase ITCH was significantly down-regulated(P<0.05).Co-immunoprecipitation confirmed the interaction between ITCH and TXNIP proteins in the brain of 2-and 4-month-old 5×FAD mice,which exhibited marked attenuation by 4 months of age.In BV2 microglial models,Aβ/LPS stimulation provoked significant ITCH suppression,concurrently up-regulating TXNIP,core NLRP3 inflammasome components(NLRP3 and caspase-1),and downstream IL-1β(P<0.05).Overexpression of ITCH significantly inhibited Aβ-induced activation of TXNIP and NLRP3 and therelated inflammatory factors in BV2 cells.CONCLUSION:The results of in vitro and in vivo experiments showed that ITCH protein exerts effects against AD-like pathology by inhibiting the expression of TXNIP-NLRP3 signaling pathway.

Objective:To analyze the epidemiological characteristics and trend of acute hemorrhagic conjunctivitis (AHC) in China from 2015 to 2022, and to provide evidence for the adjustment of prevention and control strategies and measures for AHC.Methods:The case data of AHC reported by national notifiable disease information system from 2015 to 2022 were collected, and descriptive analysis method were used to analyze the population distribution characteristics, temporal epidemiological trends and spatial clusters of AHC in China.Results:From 2015 to 2022, the incidence of AHC in China ranged from 1.85/100 000 to 2.97/100 000, with a fluctuating downward trend. The average annual percent change (AAPC) was -4.91 (95% CI: -7.74 to -2.00, P<0.05), with an annual percent change (APC) of 2.73 (95% CI: -2.34 to 8.06, P=0.189) for 2015—2019 and an APC of -14.23 (95% CI: -21.78 to -5.94, P<0.05). The age-specific incidence rate was highest in children aged 0-4 years (fluctuating between 4.69 and 5.67/100 000 from 2015 to 2019; It decreased significantly during 2020—2022, fluctuating between 1.93 and 2.72 per 100 000).The proportion of cases in children at 0-4 years of age showed a fluctuating downward trend from 8.68% in 2015 to 3.76% in 2020, with an increase in 2021—2022 to 5.74%. After 2020, the proportion of the population aged 60 years and above has increased, reaching 33.59% in 2022. Cases were mainly farmers, with a fluctuating upward trend of around 50% per year, with the highest percentage of 60.96% in 2020. The peak seasonal incidence of AHC was obvious from May to September from 2015 to 2019, but it was not obvious in 2020—2022. The cases were mainly distributed in Guangxi, Hainan and other provinces in the southwestern part of China. The high incidence counties were concentrated in Leye County of Guangxi, Maojian District of Hubei, Fuchuan Yao Autonomous County of Guangxi, Funing County of Yunnan, and Pulan County of Tibet every year. Conclusions:The overall epidemic rate of AHC in China showed a fluctuating downward trend from 2015 to 2022, with a pronounced decline observed between 2020 and 2022, potentially linked to non-pharmaceutical interventions during the COVID-19 pandemic. Increased attention needs to be given to farmers and people above 60 years of age to reduce the risk of morbidity. Moreover, prevention and control efforts should be strengthened in high-risk areas of southwestern China, and comprehensive measures should be implemented in counties with high incidence, including enhanced health education campaigns and improved allocation of sanitary facilities, to reduce the risk of AHC infection. This study is the first to highlight the potential impact of public health policies on AHC epidemiology, thereby offering a scientific foundation for population- and region-specific precision prevention strategies, particularly guiding the refinement of control measures in high-burden areas.