By selecting stable and detectable drug prototypes or metabolites in sewage samples, near real-time detection of disease conditions can be achieved. This study selected oseltamivir carboxylate, the primary metabolite of first-line antiviral oseltamivir, as a biomarker. Based on the concentration of oseltamivir carboxylate in wastewater, the consumption and usage rate of oseltamivir were calculated by reverse engineering. Quarterly sampling was conducted at 46 urban sewage treatment plants in representative 5 cities in the middle and lower reaches of the Yangtze River, from November 2022 to December 2023. The concentration range of oseltamivir acid in sewage samples is 1.270−1 279 ng/L, the daily mass load of oseltamivir per 1 000 inhabitants in the surveyed cities ranged from 9.560 to 544.7 mg/d, and the average utilization rate is 0.06‰−3.63‰. The research results indicate that in March 2023, Wuxi City experienced a spring influenza peak, while Bengbu, Tongling, Suzhou, and Changzhou City experienced a small summer influenza peak in May. In November and December 2023, Wuxi, Changzhou, and Bengbu City experienced a winter influenza peak, the results are consistent with the official statistics of the National Center for Disease Control and Prevention and the National Influenza Center, which reflect the influenza epidemic situation in southern cities. The integration of this methodology with clinical diagnostic rates could provide near real-time data support for future influenza prevention and control strategies.

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

ObjectiveTo evaluate the methodological and reporting quality of clinical practice guidelines for Chinese patent medicine （CPM） with internationally recognized tools the appraisal of guidelines for research and evaluation （AGEREE） Ⅱ and reporting items for practice guidelines in healthcare （RIGHT）， thereby providing refe-rence for the clinical application and future development of CPM guidelines. MethodsDatabases including CNKI， VIP， Wanfang and Sinomed were searched for CPM guidelines， as well as medlive.cn， websites of China Association of Chinese Medicine and Chinese Medical Association， and reference lists of the included papers. The quality of the guidelines was evaluated using the AGREE Ⅱand RIGHT tools， and consistency tests were performed using Interclass Correlation Coefficient， and descriptive analysis and chi-square test were used to analyze the reporting rate for each domain and the average score for each item. ResultsFinally， 140 CPM guidelines were included， of which 51 were disease-oriented and 89 were drug-oriented， all of which were issued by China. For 51 disease-oriented CPM guidelines， the highest average score of all six AGREE Ⅱ domains was 73.32% for clarity， and the lowest was 26.80% for application； for 89 drug-oriented CPM guidelines， the highest average score was 55.62% for scope and purpose， and the lowest was 31.32% for rigour of development. In terms of the seven domains of the RIGHT checklist， the highest reporting rate was 68.26% for background， and lowest was 27.45% for other areas regarding the disease-oriented CPM guidelines； the highest reporting rate was 61.31% for background， and the lowest was 4.49% for other areas regarding drug-oriented CPM guidelines. The average reporting rate was higher for disease-oriented than drug-oriented CPM guidelines in three domains of AGREE Ⅱ （rigour of development， clarity of presentation， editorial independence）， as well as four domains of RIGHT checklist （basic information， evidence， funding and declaration and management of interests， and other areas）. ConclusionThe overall methodology and reporting quality of the current CPM guidelines still need to be improved. It is recommended that future guideline development teams should strictly refer to the AGREE Ⅱ and RIGHT checklist， and take into account of the characteristics of CPM guidelines and relevant methodo-logical suggestions in the development and reporting of CPM guidelines， thereby guiding the clinical use of CPM in a better way.

Objective:The toll pathway plays an essential role in the inhibition of dengue virus (DENV) replication in Aedes aegypti. Accordingly, the objective of this study was to examine the impact of microRNA on the Toll pathway regulatory genes and its influence on DENV replication in Ae. aegypti. Methods:RNAhybrid software prediction and dual-luciferase reporter gene assay were employed to study the binding relationship between dps-miR-2526-3p and Rel1 A gene. The real-time fluorescence quantitative PCR (qPCR) was used in the quantification of nuclear and cytoplasmic dps-miR-2526-3p in Wolbachia-carrying (W + ) and Wolbachia-free (W -) Ae. aegypti cells via nucleocytoplasmic separation. The miRNA up-regulation assay or miRNA silencing assay combined with virus infection assay were conducted to ascertain the impact of dps-miR-2526-3p on Rel1 A gene expression and DENV replication. Results:The result of the prediction and dual-luciferase reporter gene assay consistently demonstrated the direct binding relationship between dps-miR-2526-3p and the Rel1 A gene. The qPCR result proved the presence of dps-miR-2526-3p in the nucleus and cytoplasm of Ae. aegypti cell. Notably, the expression levels of dps-miR-2526-3p and Rel1 A gene in W + cells were significantly higher than those in W -cells. The miRNA function assay indicated that dps-miR-2526-3p could positively regulate Rel1 A gene expression. The result of viral infection assay showed that dps-miR-2526-3p exhibited a notable inhibitory effect on DENV replication. Conclusions:Ae. aegypti utilizes dps-miR-2526-3p to activate the Toll pathway via Rel1 A gene, thereby inhibiting the replication of DENV. The novel molecule with anti-DENV properties was found in this study to offer a promising approach for the development of vector control strategies aimed at prevention and control of DENV transmission.

Objective:The toll pathway plays an essential role in the inhibition of dengue virus (DENV) replication in Aedes aegypti. Accordingly, the objective of this study was to examine the impact of microRNA on the Toll pathway regulatory genes and its influence on DENV replication in Ae. aegypti. Methods:RNAhybrid software prediction and dual-luciferase reporter gene assay were employed to study the binding relationship between dps-miR-2526-3p and Rel1 A gene. The real-time fluorescence quantitative PCR (qPCR) was used in the quantification of nuclear and cytoplasmic dps-miR-2526-3p in Wolbachia-carrying (W + ) and Wolbachia-free (W -) Ae. aegypti cells via nucleocytoplasmic separation. The miRNA up-regulation assay or miRNA silencing assay combined with virus infection assay were conducted to ascertain the impact of dps-miR-2526-3p on Rel1 A gene expression and DENV replication. Results:The result of the prediction and dual-luciferase reporter gene assay consistently demonstrated the direct binding relationship between dps-miR-2526-3p and the Rel1 A gene. The qPCR result proved the presence of dps-miR-2526-3p in the nucleus and cytoplasm of Ae. aegypti cell. Notably, the expression levels of dps-miR-2526-3p and Rel1 A gene in W + cells were significantly higher than those in W -cells. The miRNA function assay indicated that dps-miR-2526-3p could positively regulate Rel1 A gene expression. The result of viral infection assay showed that dps-miR-2526-3p exhibited a notable inhibitory effect on DENV replication. Conclusions:Ae. aegypti utilizes dps-miR-2526-3p to activate the Toll pathway via Rel1 A gene, thereby inhibiting the replication of DENV. The novel molecule with anti-DENV properties was found in this study to offer a promising approach for the development of vector control strategies aimed at prevention and control of DENV transmission.