Objective: To investigate the incidence, characteristics, and influencing factors of resolution discrepancies within the minipool (MP) testing model across Chinese blood station laboratories in 2024. Methods: A nationwide, multicenter, cross-sectional study was conducted, including 334 blood station laboratories that reported nucleic acid reactive data among enzyme immunoassay non-reactive samples. Of these, 296 laboratories adopted the pool resolution model, with a total of 12 536 273 samples tested. Systematic analysis was performed on resolution data, focusing on the MP-NAT reactivity rate, the pool resolution concordance rate, and the resolution discrepancy rate. Subgroup analyses were conducted based on reagent types, viral targets, and Ct values. Potential causes were further explored through laboratory surveys and re-examination of raw amplification curves. Results: In 2024, the national average MP-NAT reactivity rate was 0.15%. The overall pool resolution concordance rate was 57.86%, which showed a gradual decline as Ct values increased across all reagents. The national average resolution discrepancy rate was 0.081‱(102/12 536 273), with 17.91%(53/296) of laboratories reporting at least one discrepancy. Nine reagent types were associated with these events, exhibiting reagent-specific patterns. For Reagent A2, the predominant discrepancy was HBV reactive pools resolving as HIV (36.36%); for Reagent D1, HBV pools frequently resolved as HCV (38.89%); and for Reagent E, the most common pattern was HIV pools resolving as HBV (48.00%). These resolution discrepancies were strongly associated with high Ct values: the median pool Ct for HBV exceeded 38, while those for HCV and HIV both exceeded 40. Investigations across 16 laboratories revealed that most discrepant samples exhibited “tailing” amplification curves, with some cases linked to cross-contamination or reagent batch-specific issues. Conclusion: While the incidence of resolution discrepancies in the MP-NAT model remains low in China, variations exist across different reagents and laboratories. These discrepancies are closely associated with low viral load, reagent performance, and laboratory operational practices.

Background: Bone-target agents (BTAs), including denosumab (DMAb), are one of the bone metastasis treatments that should continue indefinitely. However, BTAs may be interrupted in some cases. In osteoporosis, DMAb withdrawal causes a rebound effect characterized by an increased bone turnover with spine fractures and hypercalcemia; evidence of the DMAb withdrawal effect in oncology is lacking. Methods: This study aimed to identify the DMAb withdrawal effect amongst lung cancer patients treated with DMAb for bone metastases between January 2020 and December 2021. Patients who discontinued DMAb were included. Encounter notes, radiological and laboratory findings were comprehensively reviewed. Results: Thirty patients were included with a median follow-up of 21 months (interquartile range [IQR], 10-30) after DMAb discontinuation. Bisphosphonates were administered before starting DMAb in 7 patients (23.3%) and after DMAb withdrawal in 4 cases (13.3%). Three cases of DMAb withdrawal-related hypercalcemia and 3 cases of spine fractures following DMAb cessation were identified in 5 patients (16.7%), all of them were females and the median age was 65 years old (IQR, 65-70). No statistical difference in DMAb duration or number of injections was found in patients developing DMAb withdrawal-related spine fractures or hypercalcemia compared with others (binary logistic regression, p=0.688 and p=0.938, respectively). Conclusions Patients with bony-metastatic lung cancer, especially post-menopausal women, are at risk of fractures and calcium abnormalities after DMAb discontinuation, suggesting that DMAb withdrawal effect may also be present in the oncological setting. A close follow-up and careful monitoring during and after discontinuation of DMAb is necessary.

Background: Bone-target agents (BTAs), including denosumab (DMAb), are one of the bone metastasis treatments that should continue indefinitely. However, BTAs may be interrupted in some cases. In osteoporosis, DMAb withdrawal causes a rebound effect characterized by an increased bone turnover with spine fractures and hypercalcemia; evidence of the DMAb withdrawal effect in oncology is lacking. Methods: This study aimed to identify the DMAb withdrawal effect amongst lung cancer patients treated with DMAb for bone metastases between January 2020 and December 2021. Patients who discontinued DMAb were included. Encounter notes, radiological and laboratory findings were comprehensively reviewed. Results: Thirty patients were included with a median follow-up of 21 months (interquartile range [IQR], 10-30) after DMAb discontinuation. Bisphosphonates were administered before starting DMAb in 7 patients (23.3%) and after DMAb withdrawal in 4 cases (13.3%). Three cases of DMAb withdrawal-related hypercalcemia and 3 cases of spine fractures following DMAb cessation were identified in 5 patients (16.7%), all of them were females and the median age was 65 years old (IQR, 65-70). No statistical difference in DMAb duration or number of injections was found in patients developing DMAb withdrawal-related spine fractures or hypercalcemia compared with others (binary logistic regression, p=0.688 and p=0.938, respectively). Conclusions Patients with bony-metastatic lung cancer, especially post-menopausal women, are at risk of fractures and calcium abnormalities after DMAb discontinuation, suggesting that DMAb withdrawal effect may also be present in the oncological setting. A close follow-up and careful monitoring during and after discontinuation of DMAb is necessary.

Objective:To investigate the feasibility of a new clinical grading scale for assessing the degree of laxity of upper eyelid skin.Methods:From May 2022 to October 2023, the patients who underwent upper eyelid skin laxity plastic surgery in the Department of Plastic Surgery and Burns-Department of Medical Cosmetology, Northern Jiangsu People’s Hospital Affiliated to Yangzhou University, were prospectively enrolled. The degree of upper eyelid skin laxity was graded independently by three plastic surgeons who did not participate in the operation according to the grading scale and using facial photographs of the patients.The clinical grading scale categorizes the degree of skin laxity into 4 grades ranging from 0 to 3 degrees, by analyzing the position of the lower edge of the upper eyelid skin relative to the upper eyelid edge and the pupil, and the higher the grade, the more serious the degree of skin relaxation. If bilateral upper eyelid skin laxity were presented, the grade of the side with more severe laxity was taken as the grade of the upper eyelid skin laxity of the patient. The Kendall coefficient (range -1-1) of the three doctors’ grading result before and after operation was calculated respectively. The closer the value was to 1, the better the consistency of the three doctors, and the better the stability of the grading scale. The degree of upper eyelid skin laxity before and after operation was expressed as M ( Q1, Q3), and the paired Wilcoxon signed rank test was used for comparison. Results:A total of 50 female patients aged 32 to 67 years old (mean 45.3 years) who underwent bilateral upper eyelid skin laxity plastic surgery were enrolled. The Kendall coefficients of the grading result of the upper eyelid skin laxity of 50 patients by three doctors before and after operation were 0.975 and 0.882, respectively ( P<0.01). It suggested high consistency among evaluators for the same patient. 50 patients were graded as 2(1, 2) degree and 0(0, 0) degree preoperatively and postoperatively, respectively, indicating that the degree of upper eyelid skin laxity was significantly improved postoperatively. The result of Wilcoxon signed rank test showed that there were significant differences in preoperative and postoperative grading result of the three doctors ( P<0.01). Conclusion:The grading scale of upper eyelid skin laxity is objective and easy to use. Its evaluation result are stable and repeatable, and it can effectively analyze the changes of the degree of upper eyelid skin relaxation before and after surgery.

This study was aimed at exploring the interaction between the nucleoprotein(NP)of influenza A virus(IAV)and TRIM25.The physicochemical properties and protein structure of IAV NP protein were analyzed through bioinformatics methods.The interaction between IAV NP and TRIM25 proteins was simulated with molecular docking techniques,and the in-teraction sites were predicted.With the cDNA of the A/Puerto Rico/8/1934(H1N1)PR8 strain as the template,the NP pro-tein was cloned into the eukaryotic expression vector pCMV-C-Flag through PCR amplification,the eukaryotic expression re-combinant plasmid pCMV-Flag-NP was constructed,and the expression was further verified.The protein expression levels of pCMV-Flag-NP and pCMV-HA-TRIM25 were detected at various time periods.The interaction between NP protein and TRIM25 protein was verified by co-immunoprecipitation.The co-localization of NP protein and TRIM25 protein in cells was ob-served with laser confocal microscopy.Bioinformatics analysis revealed that the NP protein consists of 498 amino acids and 20 amino acids,and is an unstable hydrophilic protein.The NP protein has multiple phosphorylation sites,as well as N-glycosyla-tion and O-glycosylation sites,but no transmembrane domain or signal peptide domain.Additionally,the NP protein's second-ary structure consists of a high proportion of alpha-helices and random coils.The molecular docking prediction results indicated that IAV NP interacts with TRIM25 protein and has multiple potential interaction sites,including the 233rd alanine,234th ala-nine,236th lysine,and 440th alanine of the NP protein.After successfully constructing and expressing the IAV NP protein,we verified the interaction between IAV NP and TRIM25 protein by immunoprecipitation and laser confocal microscopy obser-vations.Our results together suggested that the structure of the IAV NP protein is closely related to its function,and its im-portance to the virus is clear.In addition,the interaction between IAV NP and TRIM25 protein may be associated with TRIM25's anti-influenza virus mechanism.Further in-depth research may provide new ideas for anti-influenza virus strategies.

Objective: To investigate the prevalence of Dengue virus (DENV) infection among voluntary blood donors in Xishuangbanna Dai Autonomous Prefecture, and to evaluate the necessity of implementing nucleic acid testing (NAT) for blood donors during the rainy season (May-October). Methods: Prior to initiating donor screening, the Xishuangbanna Central Blood Center conducted in-house validation of reagent performance and participated in external quality assessment (EQA) organized by the National Center for Clinical Laboratories (NCCL). During the surveillance period (August-October 2024), a total of 2 919 donor samples were screened using a 6-sample mini-pool NAT strategy. Daily internal quality controls were recorded. Samples that tested positive in pooled screening were deconvoluted and retested in duplicate; only those reactive in both replicate wells were sent to the NCCL for confirmatory testing. At NCCL, samples underwent re-testing using five domestic NAT reagents, as well as serological assays for NS1 antigen and DENV-specific IgG/IgM. Confirmed positive samples were further characterized by serotyping, envelope (E) gene sequencing, and phylogenetic analysis using the maximum likelihood method. Results: The DENV NAT reagent demonstrated consistent detection of 40 copies/mL controls in individual donor (ID)-NAT test (mean CT: 35.61±0.40). During the 63-day quality control monitoring, DENV detection remained stable (mean CT: 22.53±0.72). The center achieved full marks in EQA assessments for 2023 and 2024. Three reactive pools were identified in initial screening, and subsequent individual testing confirmed three DENV RNA-positive donors (sample numbers: 2401, 2402, and 2403). The confirmatory test results from NCCL were: all five NAT platforms consistently detected DENV RNA in the three samples; for serological tests, 2 samples (2402, 2403) were positive for NS1 antigen, while all three samples were negative for both IgG and IgM antibodies. DENV serotyping reagents identified DENV-2 in all cases, which were further confirmed as DENV-2 Genotype Ⅱ-Cosmopolitan by E gene sequencing. Phylogenetic analysis indicated that samples 2401 and 2402 clustered with Southeast Asian strains (Thailand/MZ636802.1, Laos/PQ775621.1), while sample 2403 closely matched a previously reported local Yunnan strain (PV544686.1). Conclusion: DENV-2 infection was detected among blood donors in Xishuangbanna during the rainy season, indicating concurrent risks of imported and local transmission. We recommend implementing pooled NAT screening for blood donors in high-risk areas during dengue epidemic seasons, along with strengthened laboratory quality control, to enhance blood safety.

[Purpose]To analyze the incidence and mortality of female breast cancer in Zhejiang cancer registration areas in 2021 and the trends from 2000 to 2021.[Methods]The data of cancer inci-dence,mortality and population data reported by cancer registration areas in Zhejiang Province from 2000 to 2021 were collected.The crude incidence/mortality rates,age-standardized rates ad-justed by Chinese standard population(ASIRC,ASMRC)and world standard population(ASIRW,ASMRW),cumulative rates(0～74 years old),truncated rates(35～64 years old),proportion of fe-male breast cancer and age-specific rate were calculated.The average annual percentage change(AAPC)of female breast cancer incidence and mortality rates in Zhejiang from 2000 to 2021 was calculated using Joinpoint software.[Results]In 2021,there were 7 262 new cases of female breast cancer in Zhejiang cancer registration areas,accounting for 12.70％of all new female can-cer cases.The crude incidence rate,ASIRC and ASIRW were 65.71/105,42.03/105 and 39.22/105,respectively.The cumulative incidence rate(0～74 years old)was 4.26％,and the truncated rate(35～64 years old)was 98.34/105.The ASIRC in urban and rural areas were 43.15/105 and 40.15/105,respectively.There were 992 deaths of female breast cancer,accounting for 6.97％of all female cancer deaths.The crude mortality rate,ASMRC and ASMRW were 8.98/105,4.62/105 and 4.47/105,respectively.The cumulative mortality rate(0～74 years old)was 0.48％,and the truncated rate(35～64 years old)was 8.89/105.The ASMRC in urban and rural areas were 4.79/105 and 4.34/105,respectively.The incidence rate of female breast cancer reached the peak at the age group of 60～64 years old,and the mortality rate reached the peak at the age group of 85 years old and above.From 2000 to 2021,the crude incidence rate,ASIRC and ASIRW of female breast cancer in Zhejiang cancer registration areas showed significant increasing trends(all P＜0.05).The crude mortality rate and ASMRW of female breast cancer also showed significant increasing trends(all P＜0.05).[Conclusion]The incidence rate of female breast cancer in Zhejiang Province was higher than the national average,while the mortality rate was lower than the national average in 2021.Both the incidence and mortality rates showed increasing trends from 2000 to 2021.Fur-thermore,distinct urban-rural disparities existed in both incidence and mortality characteristics.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective:To investigate the features of peripheral blood lymphocyte subsets in children with sepsis and evaluate the value of these cells in combination with multiple indicators in the diagnosis of pediatric sepsis.Methods:A retrospective study was conducted on 86 sepsis children and 83 children with local infection admitted to Hebei Children′s Hospital from October 2022 to October 2024. Baseline clinical data, peripheral blood lymphocyte subsets, and other laboratory indicators were compared between the two groups. The least absolute shrinkage and selection operator (LASSO) regression and multivariate logistic regression were used to evaluate the independent risk factors correlated with the initiation of sepsis. The receiver operating characteristic (ROC) curve was plotted to evaluate the value of each independent risk factor for diagnosing sepsis.Results:The thrombin time, the absolute counts of NK, CD3 + T, CD4 + T, and CD8 + T cells, the levels of IgG and IgM, and the counts of lymphocytes and platelets were lower in the sepsis children than in the children with local infection. However, the prothrombin time (PT), activated partial thromboplastin time, the levels of fibrinogen, direct bilirubin, and C-reactive protein (CRP) were higher in the sepsis group (all P<0.05). The variables screened by LASSO regression were analyzed by the multivariate logistic regression, and the results showed that PT, absolute CD4 + T cell count, and the levels of IgM and CRP were independent risk factors for sepsis. The ROC analysis indicated that the area under the ROC curve (AUC) for PT, absolute CD4 + T cell count, and IgM and CRP levels when used individually in diagnosing sepsis was 0.729, 0.593, 0.605, and 0.795, respectively. The AUC for the four indexes when used in combination for diagnosing sepsis reached 0.822, showing greater superiority over that of the single index. Conclusions:The combined measurement of PT, absolute CD4 + T cell count, and the levels of IgM and CRP can improve the diagnostic efficacy for sepsis in children. Early monitoring of these indexes facilitates the assessment of the condition in children suffering from sepsis.

Objective To compare the effects of single antiplatelet therapy(SAPT)versus dual antiplatelet therapy(DAPT)on bleeding and ischemic events in patients undergoing transcatheter aortic valve replacement(TAVR)without long-term anticoagulation indications.Methods This randomized controlled trial included 90 post-TAVR patients without anticoagulation indications,who were allocated to the SAPT group(n=46,aspirin 100 mg/d)or DAPT group(n=44,aspirin 100 mg/d+clopidogrel 75 mg/d for 3 months,followed by aspirin monotherapy).Maximum aggregation rates of platelets induced by arachidonic acid(MARAA)and adenosine diphosphate(MARADP)were measured 1,3,6,and 12 months postoperatively.Bleeding and ischemic events were recorded during the follow-up visits.Results The SAPT group exhibited significantly higher MARAA and MARADP scores at 1 and 3 months,and higher MARAA scores at 6 months compared to the DAPT group(P<0.05).At the 12-month follow-up,the SAPT group had a significantly lower inci-dence of bleeding events compared to the DAPT group(13.0%vs.31.8%,P=0.043).No statistically significant difference was observed in ischemic events between the groups(15.2%vs.11.4%,P=0.759).Conclusion For TAVR patients without anticoagulation indica-tions,SAPT significantly reduced the 1-year bleeding risk compared to DAPT,without increasing ischemic events.These findings support the safety and efficacy of SAPT after TAVR.