Objective: To explore whether the long-term and frequent use of citrate anticoagulants negatively affects the bone metabolism balance of female frequent plateletpheresis donors, so as to better protect their health. Methods: A total of 65 female plateletpheresis donors and 55 female whole-blood donors from Guangzhou Blood Center (May to December 2024) were enrolled as experimental and control groups respectively, stratified into age subgroups (18-39 years and 40-60 years). Serum levels of 25-hydroxyvitamin D [25(OH)D], procollagen type I N-terminal propeptide (PINP), osteocalcin (OC), and type I collagen carboxy-terminal telopeptide (CTX) were measured. Differences in bone metabolism markers between experimental and control groups across age subgroups were compared. ANOVA was used to analyze dose-response relationships between donation age, annual apheresis donation frequency, and biochemical indicators. Results: In the 40-60 age subgroup, 25(OH)D levels were significantly lower in the experimental group (P<0.05), exhibiting a linear increase with age and a linear decrease with annual donation frequency. No significant differences in CTX or PINP levels were observed between experimental and control groups in either age subgroup. Conclusion: High-frequency plateletpheresis donation does not disrupt bone metabolic balance in female donors. However, it is associated with reduced vitamin D levels in female donors aged >40 years, potentially increasing the risk of osteoporosis. Vitamin D supplementation is recommended for high-frequency female plateletpheresis donors in this age group.

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

Objective To establish a human luminal breast cancer stem cell(BCSC)model and investigate the inhibitory effects of astragaloside Ⅳ(AS-Ⅳ)on BCSC growth.Methods MCF-7 breast cancer cells were cultured in stem cell-specific medium to induce BCSC formation.The BCSCs were then divided into a blank control group and an AS-Ⅳ treatment group,both groups were given PBS or AS-Ⅳ treatment.Morphological changes were observed after intervention.The therapeutic efficacy of AS-Ⅳ was evaluated using 3D spheroid formation and cell viability assays.Transcriptomic profiling and gene expression analysis were performed to elucidate the underlying mechanisms.Results Compared with the MCF7 breast cancer cells,MCF7 breast cancer stem cell mammospheres exhibited accelerated growth(P＜0.01)and significantly increased expression of the stemness marker ALDH1A1(P＜0.01).Further comparison with the blank control group revealed that astragaloside Ⅳ(AS-Ⅳ)treatment significantly inhibited MCF7 breast cancer stem cell proliferation(P＜0.001)and slowed mammosphere growth(P＜0.01).Transcriptomic analysis demonstrated that differentially expressed genes(DEGs)induced by stem cell modeling and AS-Ⅳ intervention were enriched in the cellular senescence signaling pathway.AS-Ⅳ intervention substantially increased the number of SA-β-gal-positive cells(P＜0.01).RT-PCR analysis confirmed that AS-Ⅳsignificantly upregulated mRNA expression of IL-1α(P＜0.01),P21(P＜0.001),and P53(P＜0.05)in MCF7 breast cancer stem cells.Conclusion Astragaloside Ⅳ suppresses the growth of human luminal breast cancer stem cells by inducing cellular senescence.

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

【Objective】 To explore the effectiveness of using donor plasma reinfusion to flush the leukoreduction system (LRS) chamber during the final reinfusion phase with the Trima Accel automated blood collection system in preventing the reduction of CD4+ and CD8+ T lymphocytes. 【Methods】 A longitudinal and cross-sectional study was designed. CD4+ count<200 cells/μL and CD8+ count<125 cells/μL were considered as the criteria for deficiency. Eighteen first-time platelet donors were followed up. The lymphocyte count was measured at 0, 3-6 and 7-14 times of blood donation in the last 300 days. 170 healthy blood donors who have not donated blood were selected as the control group. According to the cut-off point(October 2021), 88 blood donors who mainly used automatic blood collection system to donate platelet apheresis in the last 365 days(median blood donation times ≥17.5)were divided into three groups(A, B and C)and blood samples were obtained. The time for Groups A, B and C started donating platelet apheresis were as follows: Group A: before October 2019, Group B: from October 2019 to September 2021, Group C: after October 2021. Blood samples were analyzed to obtain blood counts including CD4 + and CD8 + T lymphocytes. Blood samples were analyzed to obtain blood cell counts including CD4+ and CD8+ T lymphocytes. Through a comparative analysis, this study aimed to determine if there are any statistical differences in the detection indices between the follow-up groups with varying frequencies of blood donation, the control group, and groups A, B, and C. This approach was employed to infer the efficacy of donor plasma reinfusion in flushing the leukoreduction system (LRS) chamber for preventing the decline of CD4+ and CD8+ T lymphocytes. 【Results】 Eighteen first-time blood donors who were converted to regular platelet apheresis donors did not show a decrease of CD4 + and CD8 + T lymphocytes in the 5 th and 11 th blood donation (median number of blood donation), and there was no significant difference between the above indexes and those in the 0 th blood donation. Among the previous frequent blood donors, the CD4+ and CD8+ T lymphocyte counts in Group B and Group C are both higher than the standard value, showing no statistical difference from the control group. Among regular blood donors, the CD4+ and CD8+ T lymphocyte counts in groups B and C were higher than the criteria values, and had no statistical difference compared to the control group.The CD4+ T lymphocyte count in Group A was normal, with only one donor in Group A having a CD8+ T lymphocyte count below 125 cells/μL. This donor has donated 281 times of platelet apheresis, and the group he belongs to has started blood donation 2-21 years(median of 5 years) before the adjustment of reinfusion mode. The CD4+ and CD8+ T lymphocyte counts in Group A showed significant differences compared to the control group, with median counts (Group A/Control Group) of 359/521 and 257/372, respectively, P<0.001. In Group A, 0%(0/35) had a CD4+ count below 200 cells/μL, and 2.85%(1/35) of donors had a CD8+ count below 125 cells/μL, which was far lower than the proportion of CD4+ and CD8+ T cell deficiency found in regular apheresis donors by John M. Gansner and Mahboubeh Rahmani. The study showed that the adjustment of the plasma reinfusion mode did not further reduce the T lymphocyte counts in blood donors, but instead further restored the T lymphocyte counts in regular blood donors. This indicated that after the adjustment of plasma reinfusion mode, blood donors might not have lost CD4+ and CD8+ T lymphocytes during blood donation, or only lost a small amount, and can recover even if they donate platelet apheresis frequently. 【Conclusion】 Trima Accel automated blood collection system has a good effect on preventing CD4 + and CD8 + T lymphocytes from being reduced by flushing the LRS chamber with donor plasma.

Objective: To observe the effects of apocynum venetum leaves extract(AVLE) on anxiety-like behavior caused by chronic restraint stress(CRS). Methods: Male C57BL/6 mice were randomly divided into 4 groups, 5 per group: control group(equivalent physiological saline 0.2 ml/10 g), CRS model group(equivalent physiological saline 0.2 ml/10 g), AVLE treatment group(AVLE 120 mg/kg) and positive control group(paroxetine hydrochloride 3 mg/kg). The anxiety mouse model of chronic restraint stress was established, and the anxiety level of mice treated with apocynum venetum leaf extract was evaluated by open field test and elevated cross maze test. Immunofluorescence staining was used to detect the changes of serotonin receptor in hypothalamic paraventricular nucleus. Chemogenetics combined with adeno-associated virus was used to detect the effect of hypothalamic oxytocin neurons on the anxiety regulation of AVLE. Results: Compared with the model group, apocynum venetum leaves extract could increase the total distance of open field exercise, the activity time, the time of mice entering the central area(P<0.05), and the activity distance and time of mouse elevated maze(P<0.05). The expression of serotonin 1A receptor in the paraventricular nucleus of hypothalamus was enhanced after treatment with AVLE, and inhibition of serotonin 1A receptor and oxytocin neurons in hypothalamic paraventricular nucleus could decrease activity level in mice(P<0.05). Conclusion AVLE can improve the anxiety state and activity ability of CRS mice, and its effect is related to serotonin receptor and oxytocin neurons in hypothalamic paraventricular nucleus.

Objective:To observe and analyze the surface electromyography activity of quadriceps femoris of knee osteoar-thritis(KOA)patients. Method:The surface electromyography of vastus lateralis,rectus femoris,and vastus medialis were recorded and analyzed in 30 KOA patients and 30 normal people during isokinetic knee flexion or extension at veloci-ties of 60 °/s,90 °/s or 180 °/s.We also record clinical indicators(muscle thickness and degree of knee pain)of the KOA group and evaluate the correlation between electromyographic data and clinical indicators. Result:Compared with the healthy control group,the KOA group had significantly lower knee extensor mo-ment during isokinetic knee flexion or extension at velocities of 60°/s(P＜0.001),90°/s(P＜0.01),or 180°/s(P＜0.01).Statistics showed that there were significant differences in the median frequency difference rate of vastus lateralis(P＜0.05)and root mean square value of rectus femoris(P＜0.01)between the healthy control group and the KOA group under 60°/s angular velocities.There was a significant difference in the root mean square value of rectus femoris(P＜0.05)between the two groups at velocities of 90°/s.Statistics revealed that the median frequency difference rate(P＜0.05)and root mean square value(P＜0.05)of rectus femoris had changed significantly at the velocities of 180°/s.The results of both multiple regression and random forest algo-rithm showed that knee extensor moment was the strongest predictor between electromyographic characteristics and clinical indicators. Conclusion:The strength of quadriceps femoris in KOA patients was decreased.Thus,the patient may need more muscles to complete the same action,which was easy to cause muscle fatigue.The knee extensor moment was closely related to the quadriceps femoris thickness and the degree of knee pain.It offers important advantag-es for the diagnosis and assessing the severity,which may provide the way for the future study of KOA.

Cancer has become a major life-threatening disease globally.In recent years,precision therapy for cancer has gradually emerged,and significant success has been achieved with targeted therapies,immune checkpoint inhibitors,and the newly emerging antibody-drug conjugates(ADC).The development of ADC has been rapid,with up to 15 ADC drugs currently approved worldwide.However,the hepatotoxicity of ADC has gained increasing attention,with reports of fatal hepatotoxic events.In this article,we briefly reviewed the ADC approved globally,summarized the liver-related adverse events and hepatotoxic manifestations reported in current clinical trials of ADC,and compiled research on possible hepatotoxicity mechanisms.Our aim is to assist clinicians in understanding and managing the hepatotoxicity characteristics of ADC,enabling timely identification of patients at risk of hepatotoxicity and implementing effective measures to manage these risks,thus reducing and preventing serious adverse events.

Objective:To develop a health behavior guidance program for stroke rehabilitation patients based on the adaptive leadership framework (ALF) .Methods:Relevant stroke health behavior guidelines were systematically retrieved, screened, included, and recommendations extracted from domestic and international websites. The initial draft of the health behavior guidance program for stroke rehabilitation patients was developed in light of the ALF. From January to March 2022, the draft was refined through two rounds of Delphi expert consultation, resulting in the finalized health behavior guidance program.Results:A total of 10 guidelines were included, and 73 guideline recommendations were extracted. These recommendations were categorized into six dimensions and 25 specific items. In both rounds of expert consultation, 15 questionnaires were distributed and all were returned, yielding an effective recovery rate of 100.00%, with an expert authority coefficient greater than 0.8. Based on expert feedback and group discussions, the final program included eight dimensions and 31 items, adding sleep management and regular health consultations to the original six dimensions.Conclusions:The health behavior guidance program for stroke rehabilitation patients, based on the ALF, introduces a new perspective on nursing by reasonably dividing adaptive and technical problems. This approach aligns with clinical practice in China, providing scientific and practical value for promoting health behaviors and offering a new perspective for health education in stroke patients.

Cancer has become a major life-threatening disease globally.In recent years,precision therapy for cancer has gradually emerged,and significant success has been achieved with targeted therapies,immune checkpoint inhibitors,and the newly emerging antibody-drug conjugates(ADC).The development of ADC has been rapid,with up to 15 ADC drugs currently approved worldwide.However,the hepatotoxicity of ADC has gained increasing attention,with reports of fatal hepatotoxic events.In this article,we briefly reviewed the ADC approved globally,summarized the liver-related adverse events and hepatotoxic manifestations reported in current clinical trials of ADC,and compiled research on possible hepatotoxicity mechanisms.Our aim is to assist clinicians in understanding and managing the hepatotoxicity characteristics of ADC,enabling timely identification of patients at risk of hepatotoxicity and implementing effective measures to manage these risks,thus reducing and preventing serious adverse events.