Wastewater-based epidemiology has emerged as a transformative surveillance tool for estimating substance consumption and monitoring disease prevalence, particularly during the COVID-19 pandemic. It enables the population-level monitoring of illicit drug use, pathogen prevalence, and environmental pollutant exposure. In this perspective, we summarize the key challenges specific to the Chinese context: (1) Sampling inconsistencies, necessitating standardized 24-hour composite protocols with high-frequency autosamplers (≤ 15 min/event) to improve the representativeness of samples; (2) Biomarker validation, requiring rigorous assessment of excretion profiles and in-sewer stability; (3) Analytical method disparities, demanding inter-laboratory proficiency testing and the development of automated pretreatment instruments; (4) Catchment population dynamics, reducing estimation uncertainties through mobile phone data, flow-based models, or hydrochemical parameters; and (5) Ethical and data management concerns, including privacy risks for small communities, mitigated through data de-identification and tiered reporting platforms. To address these challenges, we propose an integrated framework that features adaptive sampling networks, multi-scale wastewater sample banks, biomarker databases with multidimensional metadata, and intelligent data dashboards. In summary, wastewater-based epidemiology offers unparalleled scalability for equitable health surveillance and can improve the health of the entire population by providing timely and objective information to guide the development of targeted policies.
China/epidemiology* ; Humans ; Wastewater/analysis* ; COVID-19/epidemiology* ; Public Health ; Wastewater-Based Epidemiological Monitoring ; SARS-CoV-2

Objective:To analyze the clinical data of patients with severe fever with thrombocytopenia syndrome（SFTS）in Linyi city of Shandong province from 2023 to 2024，analyze the related factors affecting the prognosis，improve the understanding of the disease and reduce its mortality..Methods:The data of 36 SFTS patients diagnosed and admitted to Linyi People's Hospital from May 2023 to August 2024 were retrospectively analyzed. According to the clinical outcomes of the patients，they were divided into a survival group（n=30）and a death group（n=6）. The clinical data and laboratory test results of the two groups were analyzed to evaluate the risk factors related to prognosis.Results:The median age in the death group was 68.33 years old，and that in the survival group was 63.96 years old，with no statistically significant difference. All patients had fever，22 had fatigue and poor appetite，and 21 had muscle aches and other systemic symptoms. some were prone to general symptoms such as fatigue，poor appetite，and muscle soreness. All patients in the death group had neurological symptoms such as headache and consciousness disorders. The levels of serum potassium，CRP，PCT，IL-6，ALT，AST，CK，CK-MB，LDH，α-HBDH，APTT，D-D，and viral load in the death group were higher than those in the survival group，with statistically significant differences（ t=-3.344， P=0.002； Z=-2.195， P=0.028； Z=-3.648， P=0.000； Z=-3.641， P=0.000； Z=-2.241， P=0.025； Z=-2.288， P=0.022； Z=-2.427， P=0.015； Z=-2.007， P=0.045； Z=-3.127， P=0.002； Z=-2.404， P=0.016； Z=-2.755， P=0.006； Z=-3.081， P=0.002； P<0.05）. The platelet count in the death group was lower than that in the survival group，and the difference was statistically significant（ Z=-3.292， P=0.001， P<0.05）. Multivariate Logistic regression analysis showed that patients with increased AST，CK，IL-6，APTT，D-dimer and decreased platelet count had an increased risk of death. Fungal infections occurred in 15 cases，including 5 cases of Candida albicans，3 cases of Candida parapsilosis，and 7 cases of Aspergillus. All patients in the death group had fungal infections，all of whom had Aspergillus infections. Conclusion:SFTS patients often have fever，fatigue，and muscle soreness，and critically ill patients are prone to neurological symptoms. Patients with elevated AST，CK，IL-6，APTT，D-D，viral load，and decreased platelet count in the course of the disease often indicate poor prognosis and should be closely monitored. In addition，critically ill patients are prone to fungal infection.

AIM To prepare the sustained-release microspheres of ginsenosides.METHODS The sustained-release microspheres were prepared by SPG membrane emulsification technology with poly(lactic-co-glycolic acid)(PLGA)as a shell carrier.With PLGA concentration,feed rate and Span 60 concentration as influencing factors,comprehensive score for appearance,drug loading and encapsulation efficiency as an evaluation indice,the preparation process was optimized by response surface method.The morphology of sustained-release microspheres was observed,after which the particle size,drug loading and encapsulation efficiency were determined,and the in vitro drug release was investigated.RESULTS The optimal conditons were determined to be 45 s for agitation time of primary emulsion,74.68 mg/mL for PLGA concentration,11％for feed rate,and 4.18 mg/mL for Span 60 concentration,the comprehensive score was 74.98.The round sustained-release microspheres demonstrated the average particle size of 4.33 μm,drug loading of(8.24±0.13)％,and encapsulation efficiency of(74.94±1.17)％,respectively.At 336 h,ginsenosides Rg1,Rb1,Rb2 displayed the accumulative release rates of 84.12％,78.04％,65.88％,respectively.CONCLUSION This reasonable and feasible method can be used for the preparation of sustained-release microspheres of ginsenosides with good appearance and high drug loading,which can provide references for the preparation of other water-soluble drug microspheres and solution of microsphere collapse problem.

Objective To establish a highly sensitive and specific method for detecting human DNA based on real time quantitative PCR(qPCR)technique for the rapid detection of potential DNA con-tamination sources in DNA laboratories.Methods Primers and probes were designed with Primer Ex-pressTM software using the reference sequence of human 18S rRNA gene as a template,and the opti-mal prime-probe combination was screened by matrix method.The PCR products of the target se-quence of human 18S rRNA gene were used to construct the plasmid,and a plasmid standard was used to draw the standard curve of the qPCR system.According to the Minimum Information for Pub-lication of Quantitative Real-time PCR Experiments(MIQE)guidelines,the specificity,sensitivity,re-peatability and application effect of the qPCR system were evaluated.Results The sensitivity of the qPCR system established in this study was 5.3×10-5 ng/μL,which showed good specificity for human DNA samples.The correlation coefficient of the qPCR system was-0.999,and amplification efficiency was 100％.Both the intra-batch and inter-batch variation coefficients were less than 2％.Conclusion The established human DNA detection method based on qPCR technique has good specificity,high sen-sitivity,and robust stability.It can be used for rapid detection of DNA contamination and daily moni-toring of the accumulated human DNA in the laboratory environment.

Objective To explore the application of plasma 7 microRNA (miR7) testing in the differential diagnosis of very early-stage hepatocellular carcinoma (HCC). Methods This study is a multicenter real-world study. Patients with single hepatic lesion (maximum diameter≤2 cm) who underwent plasma miR7 testing at Zhongshan Hospital, Fudan University, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Anhui Provincial Hospital, and Peking University People’s Hospital between January 2019 and December 2024 were retrospectively enrolled. Patients were divided into very early-stage HCC group and non-HCC group, and the clinical pathological characteristics of the two groups were compared. The value of plasma miR7 levels, alpha-fetoprotein (AFP), and des-gamma-carboxy prothrombin (DCP) in the differential diagnosis of very early-stage HCC was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). In patients with both negative AFP and DCP (AFP＜20 ng/mL, DCP＜40 mAU/mL), the diagnostic value of plasma miR7 for very early-stage HCC was analyzed. Results A total of 64 528 patients from 4 hospitals underwent miR7 testing, and 1 682 were finally included, of which 1 073 were diagnosed with very early-stage HCC and 609 were diagnosed with non-HCC. The positive rate of miR7 in HCC patients was significantly higher than that in non-HCC patients (67.9% vs 24.3%, P＜0.001). ROC curves showed that the AUCs for miR7, AFP, and DCP in distinguishing HCC patients from the non-HCC individuals were 0.718, 0.682, and 0.642, respectively. The sensitivities were 67.85%, 43.71%, and 44.45%, and the specificities were 75.70%, 92.78%, and 83.91%, respectively. The pairwise comparison of AUCs showed that the diagnostic efficacy of plasma miR7 detection was significantly better than that of AFP or DCP (P＜0.05). Although its specificity was slightly lower than AFP and DCP, the sensitivity was significantly higher. Among patients negative for both AFP and DCP, miR7 maintained an AUC of 0.728 for diagnosing very early-stage HCC, with 67.82% sensitivity and 77.73% specificity. Conclusions Plasma miR7 testing is a potential molecular marker with high sensitivity and specificity for the differential diagnosis of small hepatic nodules. In patients with very early-stage HCC lacking effective molecular markers (negative for both AFP and DCP), miR7 can serve as a novel and effective molecular marker to assist diagnosis.

Preeclampsia (PE) is a severe gestational disorder characterized by hypertension and proteinuria, with a subset of cases exhibiting an immune-driven phenotype marked by placental overexpression of proinflammatory cytokines and chronic inflammatory damage, profoundly impacting fetal development. To elucidate the pathophysiology of this PE subtype, we established an inflammation-driven PE mouse model via lipopolysaccharide (LPS) intraperitoneal injection, systematically evaluating histopathological changes in maternal heart, liver, lung, kidney, and placenta, and integrating transcriptomic profiling to uncover molecular mechanisms. LPS administration robustly induced maternal hypertension and proteinuria, hallmarks of PE, without significantly altering organ or fetal weights. Histological analyses revealed pronounced inflammatory damage in the maternal lung, kidney, and placenta, with the lung exhibiting the most severe pathology, characterized by inflammatory cell infiltration, alveolar wall thickening, and interstitial edema-challenging the conventional focus on placental and renal primacy in PE. Placental labyrinth and junctional zones displayed extensive structural disruption and necrosis, indicating functional impairment. Transcriptomic analysis identified 27 inflammation-related genes consistently upregulated across tissues, with protein-protein interaction networks pinpointing Il1β, Il6, Ccl5, Ccl2, Cxcl10, Tlr2, and Icam1 as hub genes. Quantitative PCR validation confirmed Tlr2 as a central regulator, evidenced by significant upregulation of Tlr2 in lung, kidney, and placenta of LPS-induced PE mice, while Cxcl10 exhibited placenta-specific upregulation, suggesting a synergistic inflammatory axis in placental pathology. These findings highlight the lung as a critical, yet underappreciated, target in inflammation-driven PE, reframe the multi-organ inflammatory landscape of the disease, and nominate Tlr2 and Cxcl10 as potential diagnostic biomarkers and therapeutic targets, offering new avenues for precision intervention in PE.
Animals ; Female ; Pregnancy ; Mice ; Pre-Eclampsia/genetics* ; Inflammation ; Lipopolysaccharides/adverse effects* ; Disease Models, Animal ; Transcriptome ; Placenta/pathology* ; Phenotype

AIM To prepare the sustained-release microspheres of ginsenosides.METHODS The sustained-release microspheres were prepared by SPG membrane emulsification technology with poly(lactic-co-glycolic acid)(PLGA)as a shell carrier.With PLGA concentration,feed rate and Span 60 concentration as influencing factors,comprehensive score for appearance,drug loading and encapsulation efficiency as an evaluation indice,the preparation process was optimized by response surface method.The morphology of sustained-release microspheres was observed,after which the particle size,drug loading and encapsulation efficiency were determined,and the in vitro drug release was investigated.RESULTS The optimal conditons were determined to be 45 s for agitation time of primary emulsion,74.68 mg/mL for PLGA concentration,11％for feed rate,and 4.18 mg/mL for Span 60 concentration,the comprehensive score was 74.98.The round sustained-release microspheres demonstrated the average particle size of 4.33 μm,drug loading of(8.24±0.13)％,and encapsulation efficiency of(74.94±1.17)％,respectively.At 336 h,ginsenosides Rg1,Rb1,Rb2 displayed the accumulative release rates of 84.12％,78.04％,65.88％,respectively.CONCLUSION This reasonable and feasible method can be used for the preparation of sustained-release microspheres of ginsenosides with good appearance and high drug loading,which can provide references for the preparation of other water-soluble drug microspheres and solution of microsphere collapse problem.

Objective:To analyze the clinical data of patients with severe fever with thrombocytopenia syndrome（SFTS）in Linyi city of Shandong province from 2023 to 2024，analyze the related factors affecting the prognosis，improve the understanding of the disease and reduce its mortality..Methods:The data of 36 SFTS patients diagnosed and admitted to Linyi People's Hospital from May 2023 to August 2024 were retrospectively analyzed. According to the clinical outcomes of the patients，they were divided into a survival group（n=30）and a death group（n=6）. The clinical data and laboratory test results of the two groups were analyzed to evaluate the risk factors related to prognosis.Results:The median age in the death group was 68.33 years old，and that in the survival group was 63.96 years old，with no statistically significant difference. All patients had fever，22 had fatigue and poor appetite，and 21 had muscle aches and other systemic symptoms. some were prone to general symptoms such as fatigue，poor appetite，and muscle soreness. All patients in the death group had neurological symptoms such as headache and consciousness disorders. The levels of serum potassium，CRP，PCT，IL-6，ALT，AST，CK，CK-MB，LDH，α-HBDH，APTT，D-D，and viral load in the death group were higher than those in the survival group，with statistically significant differences（ t=-3.344， P=0.002； Z=-2.195， P=0.028； Z=-3.648， P=0.000； Z=-3.641， P=0.000； Z=-2.241， P=0.025； Z=-2.288， P=0.022； Z=-2.427， P=0.015； Z=-2.007， P=0.045； Z=-3.127， P=0.002； Z=-2.404， P=0.016； Z=-2.755， P=0.006； Z=-3.081， P=0.002； P<0.05）. The platelet count in the death group was lower than that in the survival group，and the difference was statistically significant（ Z=-3.292， P=0.001， P<0.05）. Multivariate Logistic regression analysis showed that patients with increased AST，CK，IL-6，APTT，D-dimer and decreased platelet count had an increased risk of death. Fungal infections occurred in 15 cases，including 5 cases of Candida albicans，3 cases of Candida parapsilosis，and 7 cases of Aspergillus. All patients in the death group had fungal infections，all of whom had Aspergillus infections. Conclusion:SFTS patients often have fever，fatigue，and muscle soreness，and critically ill patients are prone to neurological symptoms. Patients with elevated AST，CK，IL-6，APTT，D-D，viral load，and decreased platelet count in the course of the disease often indicate poor prognosis and should be closely monitored. In addition，critically ill patients are prone to fungal infection.

The ICR(Institute of Cancer Research)mouse infection model was constructed to study the pathogenicity of Sal-monella Telelkebir serotype,and the pathogenic identification of mouse isolates was carried out.Observe the bacterial excretion cycle,evaluate the pathogenicity of Salmonella serotype to mice,and calculate the LD50 by the changes in clinical characteris-tics,histopathology and tissue bacterial load of infected mice;by flight mass spectrometry,biochemical identification,serotype identification,molecular typing and other experiments,compared with human isolates;virulence gene analysis was carried out by PCR experiment and whole genome sequencing.The LD50 of Salmonella Telelkebir is 2.67 × 108 CFU/mL;curling and fluffing may occur 0.5 h after infection;autopsy of dead mice showed that the small intestine was severely congested,with more bubbles and fluid accumulation,cecal necrosis,liver apical degeneration and necrosis,necrotic foci on the surface of the kidney and spleen atrophy;the bacterial load of spleen,kidney,lung,liver and jejunum in mice reached its peak at 3 days after infection,while that of heart at 6 days;the bacterial excretion time of the high-dose group exceeded 100 days;The level of CD3 in tissues increased with increasing dose,with inflammatory cell infiltration,myocardial capillary dilation and hyperemia,large area of vacuoles,degeneration and necrosis of hepatocytes,obvious enlargement of splenic sinus,blurred zoning,thickening of glomerular basement membrane,partial exfoliation of ciliated epithelium,atrophy and exfoliation of jejunal villi;PCR and whole genome sequencing revealed Salmonella-related virulence genes such as cdtB,plt A and pltB.This study was the first to successfully establish the ICR mouse model of Salmonella Telelkebir,demonstrating that this serotype of Salmonella has some pathogenicity.