Objective:To investigate the disparities in breast cancer burden and changing epidemiological trends between China and the United States(US)from 1990 to 2021 and identify the key driving factors.Methods:The Global Burden of Disease database(1990-2021)was utilized to analyze breast cancer incidence,mortality,and disability adjusted life years(DALYs)in China and the US;we also assessed the as-sociations with risk factors(e.g.,obesity and smoking)through regression models and spatiotemporal analysis.Results:China's breast can-cer age-standardized incidence rate(ASIR)surged from 9.08/100,000 in 1990 to 19.36/100,000 in 2021,whereas the US ASIR declined from 44.37/100,000 to 37.49/100,000.Regarding the age-standardized mortality rate(ASMR),China saw a marginal reduction from 4.70/100,000 to 4.40/100,000,whereas the US experienced a significant decline from 13.03/100,000 to 9.28/100,000,narrowing the mortality gap from 2.8-fold to 2.1-fold.Key risk factors contributing to mortality and DALYs included smoking,exposure to secondhand smoke,low physical activity,high red meat and alcohol intake,elevated fasting blood glucose levels,and high body mass index.The rising cancer incidence in China is associated with the westernization of lifestyles and changes in fertility patterns,whereas the United States has achieved a dual de-cline in both incidence and mortality rates through widespread screening and control of hormone replacement therapy.Conclusions:Both the incidence and mortality rates of breast cancer in China rank among the highest globally,with incidence rates still increasing,indicating severe issues in prevention and control.In contrast,although the incidence rate in the US is higher than that in China,it shows a declining trend.To alleviate the growing burden of breast cancer,China must further enhance the intensity and coverage of breast cancer screening.

Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45％.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3％(GyVg1)and 100％(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.

To establish a double fluorescent RT-LAMP that can distinguish between chicken astro-virus(CAstV)and avian nephritis virus(ANV),two sets of specific primers and probes labeled with fluorescent groups were designed by comparing the conserved sequences of CAstV ORF1b gene and ANV ORF1b gene downloaded from GenBank.The CAstV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The ANV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The reaction system of the method was optimized,and specificity,repeatability,interference,sensitivity,and clinical sample detection were performed.The results were compared with RT-qPCR and RT-PCR to verify the ef-fectiveness of the method.The experimental results showed that the established double fluorescence RT-LAMP method could complete the reaction within 60 min,and the optimal reac-tion temperature was 65 ℃.This method only detects ANV and CAstV,with no cross-reactivity to other common avian pathogens.The minimum detection limit of CAstV was 1.4 ×10 2 copies/μL,and the minimum detection limit of ANV was 1.5×10 2 copies/μL.The interference and reproduc-ibility tests were good.Compared with RT-qPCR,the coincidence rates of CAstV and ANV were 97.97％and 98.85％,respectively.In conclusion,the double fluorescence RT-LAMP method estab-lished in this study has the characteristics of rapidity,accuracy,good specificity,high sensitivity and good reproducibility,which is suitable for clinical diagnosis and provides technical support for the epidemiological investigation of CAstV and ANV.

Objective:To investigate the disparities in breast cancer burden and changing epidemiological trends between China and the United States(US)from 1990 to 2021 and identify the key driving factors.Methods:The Global Burden of Disease database(1990-2021)was utilized to analyze breast cancer incidence,mortality,and disability adjusted life years(DALYs)in China and the US;we also assessed the as-sociations with risk factors(e.g.,obesity and smoking)through regression models and spatiotemporal analysis.Results:China's breast can-cer age-standardized incidence rate(ASIR)surged from 9.08/100,000 in 1990 to 19.36/100,000 in 2021,whereas the US ASIR declined from 44.37/100,000 to 37.49/100,000.Regarding the age-standardized mortality rate(ASMR),China saw a marginal reduction from 4.70/100,000 to 4.40/100,000,whereas the US experienced a significant decline from 13.03/100,000 to 9.28/100,000,narrowing the mortality gap from 2.8-fold to 2.1-fold.Key risk factors contributing to mortality and DALYs included smoking,exposure to secondhand smoke,low physical activity,high red meat and alcohol intake,elevated fasting blood glucose levels,and high body mass index.The rising cancer incidence in China is associated with the westernization of lifestyles and changes in fertility patterns,whereas the United States has achieved a dual de-cline in both incidence and mortality rates through widespread screening and control of hormone replacement therapy.Conclusions:Both the incidence and mortality rates of breast cancer in China rank among the highest globally,with incidence rates still increasing,indicating severe issues in prevention and control.In contrast,although the incidence rate in the US is higher than that in China,it shows a declining trend.To alleviate the growing burden of breast cancer,China must further enhance the intensity and coverage of breast cancer screening.

Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45％.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3％(GyVg1)and 100％(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.

To establish a double fluorescent RT-LAMP that can distinguish between chicken astro-virus(CAstV)and avian nephritis virus(ANV),two sets of specific primers and probes labeled with fluorescent groups were designed by comparing the conserved sequences of CAstV ORF1b gene and ANV ORF1b gene downloaded from GenBank.The CAstV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The ANV probe was labeled with CY3 fluorophore at the 3'end and the quencher BHQ2 at the 5'end.The reaction system of the method was optimized,and specificity,repeatability,interference,sensitivity,and clinical sample detection were performed.The results were compared with RT-qPCR and RT-PCR to verify the ef-fectiveness of the method.The experimental results showed that the established double fluorescence RT-LAMP method could complete the reaction within 60 min,and the optimal reac-tion temperature was 65 ℃.This method only detects ANV and CAstV,with no cross-reactivity to other common avian pathogens.The minimum detection limit of CAstV was 1.4 ×10 2 copies/μL,and the minimum detection limit of ANV was 1.5×10 2 copies/μL.The interference and reproduc-ibility tests were good.Compared with RT-qPCR,the coincidence rates of CAstV and ANV were 97.97％and 98.85％,respectively.In conclusion,the double fluorescence RT-LAMP method estab-lished in this study has the characteristics of rapidity,accuracy,good specificity,high sensitivity and good reproducibility,which is suitable for clinical diagnosis and provides technical support for the epidemiological investigation of CAstV and ANV.

The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P＜0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P＞0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.

Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.

Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.

Background: Chicken anemia virus (CAV) causes chicken infectious anemia, which results in immunosuppression; the virus has spread widely in chicken flocks in China. Objectives: The aim of this study was to understand recent CAV genetic evolution in chicken flocks in Guangxi Province, southern China. Methods: In total, 350 liver samples were collected from eight commercial broiler chicken farms in Guangxi Province in southern China from 2018 to 2020. CAV was detected by conventional PCR, and twenty CAV complete genomes were amplified and used for the phylogenetic analysis and recombination analysis. Results: The overall CAV-positive rate was 17.1%. The genetic analysis revealed that 84 CAVs were distributed in groups A, B, C (subgroups C1-C3) and D. In total, 30 of 47 Chinese CAV sequences from 2005-2020 belong to subgroup C3, including 15 CAVs from this study. There were some specific mutation sites among the intergenotypes in the VP1 protein. The amino acids at position 394Q in the VP1 protein of 20 CAV strains were consistent with the characteristics of a highly pathogenic strain. GX1904B was a putative recombinant. Conclusions Subgroup C3 was the dominant genotype in Guangxi Province from 2018–2020.The 20 CAV strains in this study might be virulent according to the amino acid residue analysis. These data help improve our understanding of the epidemiological trends of CAV in southern China.