Objectives:To observe the mitochondrial morphology of normal and triple-negative breast cancer cells, extract mitochondria from normal cells, and investigate the effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of triple-negative breast cancer cells.Methods:The morphology of mitochondria was observed by transmission electron microscope. Mitochondria were extracted by mitochondrial extraction kit, mitochondrial protein was identified by western blot, and mitochondrial activity was detected by mitochondrial membrane potential detection kit. MitoTracker Green or MitoTracker Deep Red fluorescent probes were used to label the mitochondria of living cells, and the degree of mitochondria entering LTT cells was observed by confocal laser microscopy at 12, 24, and 96 hours. The effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of breast cancer cells were examined by CCK8, colony formation assay, flow cytometry, and sphere formation assay after 24 hours of mitochondrial transplantation.Results:The mitochondria of normal cells were rod-shaped or elongated, while the mitochondria of triple-negative breast cancer cells were swollen and vacuolated. Western blot results showed that cytochrome c oxidase subunit I (MT-CO1) protein encoded by mitochondria was present in the isolated mitochondria. The content of heat shock protein 60 (HSP60) was higher in mitochondria than that in cytoplasm. The result of the multi-mode microplate reader showed that the content of mitochondrial J-aggregates/monomer was 1.67±0.06, which was significantly higher than 0.35±0.04 of the control group ( P＜0.001). Exogenous mitochondria were observed in LTT cells at 12, 24, and 96 hours after mitochondrial transplantation. The results of the CCK8 experiment showed that OD450 of LTT cells was 0.27±0.13 after 48 hours transplantation, which was lower than 0.62±0.36 of the control group ( P=0.023). The OD450 of MDA-MB-468 cells was 0.30±0.03, which was lower than 0.65±0.10 of the control group ( P=0.004). After 120 hours of mitochondrial transplantation, OD450 in both groups was still significantly lower than that in the control group (P＜0.01). The number of clones formed by mitochondrial transplantation of LTT cells was 21.33±7.31, which was lower than 35.22±13.59 of the control group ( P=0.016). Flow cytometry showed that the early apoptosis rate of LTT cells was (30.07±2.15)% after 24 hours of mitochondrial transplantation, which was higher than 2.07±1.58 of the control group ( P＜0.001). The proportion of early apoptosis in MDA-MB-468 cells was 24.47%±5.22%, which was higher than (7.83±2.06)% in the control group ( P=0.007). In addition, the number of mitochondria transplanted LTT cells into the cell sphere was 46.25±5.40, which was significantly lower than 62.58±6.43 of the control group ( P＜0.001). Conclusion:Normal mitochondria can enter triple-negative breast cancer cells by co-culture, inhibit the proliferation and stemness of triple-negative breast cancer cells, and promote the apoptosis of triple-negative breast cancer cells.

BACKGROUND:Red light irradiation and silver ion dressing are mostly used to treat chronic difficult healing wounds clinically,but the optimal irradiation time of red light irradiation and silver ion dressing for chronic non-healing wounds,and the combination of different silver ion dressings have not been determined. OBJECTIVE:To investigate the optimal irradiation time and dressing combination of red light and silver ion dressing in the therapy of chronic non-healing wounds. METHODS:The chronic non-healing wound model was made by applying Staphylococcus aureus on the whole skin defect and subcutaneous hydrocortisone injection in SD rats.72 rat models were randomly divided into 4 groups with 18 rats in each group by random number table method.The rats were treated on the basis of standard dressing change and the following therapy:A1B1 group(red irradiation 20 minutes + lipid hydrocolloidal silver sulfate dressing),A1B2 group(red light irradiation 20 minutes + calcium alginate fiber dressing),A2B1 group(red light irradiation 30 minutes + lipid hydrocolloidal silver sulfate dressing),and A2B2 group(red light irradiation 30 minutes + calcium alginate fiber dressing);change dressing,irradiate once,and change dressing every 24 hours.After 14 days of continuous treatment,wound healing rate,bacterial colony number,inflammatory response,histomorphology and angiogenesis were detected in each group. RESULTS AND CONCLUSION:(1)With the extension of treatment time,the wound healing rate of rats in the four groups was increased,and the wound healing rate of rats in the A2B2 group at 3,7,and 14 days after treatment was higher than that in the other three groups(P<0.05).(2)The wound bacterial culture results on day 7 after treatment demonstrated that the number of bacterial colonies in the A2B2 group was lower than that in the other three groups(P<0.05).Western blot assay exhibited that with the extension of treatment time,the protein expressions of tumor necrosis factor α and interleukin-6 in wound tissue of rats in the four groups were decreased,while the protein expressions of interleukin-10 were increased.The protein expressions of tumor necrosis factor α and interleukin-6 in the A2B2 group were lower than those in the other three groups(P<0.05).The protein expression of interleukin-10 in the A2B2 group was higher than that of the other three groups(P<0.05).(3)The wound hematoxylin-eosin staining on day 14 after treatment demonstrated that a large number of collagen fibers in the A2B2 group were parallel distributed and the most closely connected,which was significantly better than the other three groups.(4)The results of immunofluorescence staining indicated that the fluorescence intensity expression of CD31 in the A2B2 group was higher than that in the A1B1,A1B2 and A2B1 groups(P<0.05).q-PCR detection at 3,7,and 14 days after treatment exhibited that the mRNA expressions of vascular endothelial growth factor a and vascular endothelial growth factor receptor 2 in the A2B2 group were higher than those in the other three groups(P<0.05).Western blot assay at 3,7 and 14 days after treatment revealed that the protein expressions of vascular endothelial growth factor a and vascular endothelial growth factor receptor 2 in the A2B2 group were higher than those in the other three groups(P<0.05).(5)These findings confirm that 30 minutes of red light irradiation combined with silver alginate fiber dressing has better results in treatment of chronic non-healing wounds.

Objectives:To observe the mitochondrial morphology of normal and triple-negative breast cancer cells, extract mitochondria from normal cells, and investigate the effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of triple-negative breast cancer cells.Methods:The morphology of mitochondria was observed by transmission electron microscope. Mitochondria were extracted by mitochondrial extraction kit, mitochondrial protein was identified by western blot, and mitochondrial activity was detected by mitochondrial membrane potential detection kit. MitoTracker Green or MitoTracker Deep Red fluorescent probes were used to label the mitochondria of living cells, and the degree of mitochondria entering LTT cells was observed by confocal laser microscopy at 12, 24, and 96 hours. The effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of breast cancer cells were examined by CCK8, colony formation assay, flow cytometry, and sphere formation assay after 24 hours of mitochondrial transplantation.Results:The mitochondria of normal cells were rod-shaped or elongated, while the mitochondria of triple-negative breast cancer cells were swollen and vacuolated. Western blot results showed that cytochrome c oxidase subunit I (MT-CO1) protein encoded by mitochondria was present in the isolated mitochondria. The content of heat shock protein 60 (HSP60) was higher in mitochondria than that in cytoplasm. The result of the multi-mode microplate reader showed that the content of mitochondrial J-aggregates/monomer was 1.67±0.06, which was significantly higher than 0.35±0.04 of the control group ( P＜0.001). Exogenous mitochondria were observed in LTT cells at 12, 24, and 96 hours after mitochondrial transplantation. The results of the CCK8 experiment showed that OD450 of LTT cells was 0.27±0.13 after 48 hours transplantation, which was lower than 0.62±0.36 of the control group ( P=0.023). The OD450 of MDA-MB-468 cells was 0.30±0.03, which was lower than 0.65±0.10 of the control group ( P=0.004). After 120 hours of mitochondrial transplantation, OD450 in both groups was still significantly lower than that in the control group (P＜0.01). The number of clones formed by mitochondrial transplantation of LTT cells was 21.33±7.31, which was lower than 35.22±13.59 of the control group ( P=0.016). Flow cytometry showed that the early apoptosis rate of LTT cells was (30.07±2.15)% after 24 hours of mitochondrial transplantation, which was higher than 2.07±1.58 of the control group ( P＜0.001). The proportion of early apoptosis in MDA-MB-468 cells was 24.47%±5.22%, which was higher than (7.83±2.06)% in the control group ( P=0.007). In addition, the number of mitochondria transplanted LTT cells into the cell sphere was 46.25±5.40, which was significantly lower than 62.58±6.43 of the control group ( P＜0.001). Conclusion:Normal mitochondria can enter triple-negative breast cancer cells by co-culture, inhibit the proliferation and stemness of triple-negative breast cancer cells, and promote the apoptosis of triple-negative breast cancer cells.

Objective:To investigate the clinical efficacy of non-drug interventions in hospice and palliative care(HPC)for improving cancer-related fatigue(CRF)in elderly individuals and its impact on quality of life.Methods:This study presents findings from a single-center randomized controlled trial conducted at Zhejiang Hospital, focusing on 40 elderly patients experiencing cancer-related fatigue(CRF)between February 2022 and February 2023.The participants were randomly assigned into a control group and an intervention group, each consisting of 20 individuals, using the random number table method.Both groups received routine comprehensive treatment, with the intervention group additionally receiving hospice and palliative care(HPC)non-drug intervention.Following 6 weeks of continuous treatment, the study compared the clinical efficacy, changes in CRF, and quality of life before and after treatment between the two groups.Results:Comparing the baseline data of the two groups of patients, the difference was not statistically significant(all P>0.05).After 6 weeks of treatment, patients in the intervention group reported lower levels of current fatigue, general fatigue, worst fatigue in the past 24 hours, and impact of fatigue on various aspects of their lives compared to the control group(all P<0.01).The clinical remission rate of cancer-related fatigue(CRF)in the intervention group was 60%, significantly higher than the 5% in the control group( P<0.01).Additionally, the intervention group showed improvement in overall quality of life and emotional function with decreased symptom areas scores(fatigue, nausea and vomiting, shortness of breath, sleep disorders, and loss of appetite)( P<0.01 for quality of life and emotional function, P<0.05 for symptom areas). Conclusions:Non-pharmacological interventions within the context of hospice and palliative care have been shown to alleviate cancer-related fatigue in elderly cancer patients, ultimately improving their quality of life.These interventions have demonstrated positive effects on various aspects such as overall quality of life, functional status, fatigue, nausea and vomiting, shortness of breath, sleep disorders, and decreased appetite.Furthermore, these interventions are considered safe and effective in the treatment of elderly cancer patients experiencing fatigue.

[摘 要] 目的：探索抗HSP90单克隆抗体28C10通过靶向肿瘤干细胞促进顺铂（cisplatin，DDP）对人胃癌细胞PAMC82恶性生物学行为的抑制效果及其可能的作用机制。方法: 28C10单独或与DDP联合处理人胃癌细胞PAMC82，采用不同实验方法检测该细胞的无血清成球能力、迁移和侵袭能力与克隆形成能力，CCK-8法检测28C10对PAMC82细胞恶性生物学行为和协同DDP抗癌能力的影响。采用细胞免疫荧光及流式细胞术检测PAMC82细胞中HSP90及eHSP90（extracellular HSP90）的表达、定位、eHSP90+亚群比例，以及28C10处理后对ALDH+、CD44+、eHSP90+细胞亚群的影响。采用WB实验检测28C10作用后PAMC82细胞中HSP90、干性相关蛋白以及PI3K/AKT/mTOR信号通路蛋白表达的变化。结果：胃癌细胞PAMC82膜表面表达eHSP90，具有2%~3%的eHSP90+细胞亚群，且eHSP90+细胞多为与ALDH+或CD44+共阳性细胞。28C10处理能显著抑制PAMC82细胞的成球、克隆形成、增殖、耐药、迁移及侵袭能力，而且和DDP联用的效果更明显（P<0.05或P<0.01）。流式细胞术分析发现28C10处理显著抑制PAMC82细胞的eHSP90+、ALDH+和CD44+亚群数量（均P<0.01）。免疫荧光实验发现28C10作用后eHSP90发生内吞，WB实验结果显示eHSP90、CD44、ALDH和干性相关蛋白OCT4、SOX2表达量均降低（P<0.05或P<0.01）。结论：抗HSP90单克隆抗体28C10可靶向胃癌PAMC82细胞的ALDH+、CD44+肿瘤干细胞相关亚群、内化eHSP90且降低细胞总HSP90的水平、抑制PI3K/AKT/mTOR信号通路，从而有效地抑制PAMC82细胞的干性、耐药和其他恶性生物学行为，协同DDP显著提高抗癌效果。

Objective:To investigate the effects of propofol and sevoflurane on post-traumatic stress disorder (PTSD) after emergency surgery in trauma patients.Methods:A total of 160 trauma patients undergoing emergency surgery under general anesthesia were randomly divided into the propofol group and the sevoflurane group. The perioperative clinical data of the two groups were collected. The incidence of PTSD was evaluated by PCL-5 score one month after the operation in the two groups. The relevance of the injury time and PCL-5 score was assessed by Spearman correlation analysis. Logistic regression analysis was used to analyze the risk factors of PTSD.Results:The incidence of PTSD in the propofol group was significantly higher than that in the sevoflurane group at postoperative 1 month (24.0% vs 10.8%, P=0.034). The injury time was negatively correlated with PCL-5 score in the propofol group ( r=0.229, P<0.01). There was no correlation between the injury time and the PCL-5 score in the sevoflurane group ( r=0.001, P=0.804). Logistic regression analysis showed that the use of propofol was an independent risk factor for PTSD ( P=0.004). Conclusions:Sevoflurane anesthesia is more effective than propofol anesthesia in reducing the occurrence of PTSD in emergency surgery for trauma patients.

[Abstract] Objective: To explore the effect of anti-ENO1 (enolase 1) antibody and metformin (MET) treatment on the proliferation, migration, invasion and stemness of cetuximab (CTX) -resistant non-small cell lung cancer (NSCLC) cells through targeting cancer stem cells and the possible mechanism. Methods: 10 mmol/L MET combined with 40 μg/ml anti-ENO1 antibody was used to treat CTX(35 µg/ml)-resistant NSCLC A549 cells for 4 d, and the effects of combined treatment on A549 cells were detected with proliferation experiment, colony formation assay, migration and invasion experiments and methylcellulose ball formation experiment. In the meanwhile, FCM was used to detect the effects of CTX, MET and anti-ENO1 antibody single-drug treatment as well as the three-drug combination treatment on ALDH+ and CD44+ lung cancer stem cell subsets. Results: CTX combined with MET and anti-ENO1 antibody treatment significantly inhibited the proliferation, migration, invasion and self-renewal capacity of A549 cells. FCM analysis found that MET could significantly inhibit ALDH+ stem cell subpopulations, while anti-ENO1 antibody could significantly inhibit CD44+ stem cell subpopulations, and the three-drug combination treatment could simultaneously suppress ALDH+ and CD44+ stem cell subpopulations. Conclusion: MET and anti-ENO1 antibody respectively target ALDH+ and CD44+ cancer stem cell subsets, and the combined treatment of MET and anti-ENO1 antibody can effectively reverse the resistance of A549 cells to CTX, and thereby more effectively inhibiting stemness, proliferation, metastasis of A549 cells and tumor recurrence.

Objective:To observe the optical coherence tomography angiography (OCTA) image characteristics of polypoid choroidal vascular disease (PCV) after intravitreal injection of anti-vascular endothelial growth factor drugs, and to discuss its significance in the diagnosis and follow-up of PCV.Methods:A retrospective case study. From August 2018 to January 2020, 22 eyes of 22 patients with PCV diagnosed in the ophthalmological examination of Affiliated Hospital of Weifang Medical University were included in the study. Among them, there were 10 males with 10 eyes and 12 females with 12 eyes; the average age was 67.75±9.53 years. Best corrected visual acuity (BCVA), OCTA, and indocyanine green angiography (ICGA) were performed. All the affected eyes were injected vitreously with 10 mg/ml Conbercept 0.05 ml (including Conbercept 0.5 mg) once a month for 3 consecutive months.Tthe macular area of 3 mm×3 mm and 6 mm×6 mm with an OCTA instrument was scanned, and the foveal retinal thickness (CRT) was measured, the area of abnormal branch blood vessels (BVN). pigment epithelial detachment before and 12 months after treatment (PED) height, foveal choroid thickness (SFCT) were performed. The diagnosis rate of PCV by OCTA was observed, as well as the changes of various indicators of BCVA and OCTA. Before and after treatment, BCVA and CRT were compared by paired t test; BVN area, PED height, and SFCT were compared by variance analysis. The changes in imaging characteristics of OCTA before and after treatment were analyzed. Results:Among the 22 eyes, 8 eyes were BVN; 5 eyes were polypoid lesions (polyps); 5 eyes were BVN combined with polyps; 3 eyes were not found with BVN and polyps; 1 eye with small vascular network structure, this eye was ICGA Appears as strong nodular fluorescence (polyps). The detection rate of PCV by OCTA was 86.36% (19/22). Twelve months after treatment, BVN was significantly reduced or disappeared in 16 eyes (72.72%, 16/22); polyps disappeared in 17 eyes (77.27%, 17/22). Compared with before treatment, 12 months after treatment, BCVA increased ( t=3.071), CRT decreased ( t=2.440), the difference was statistically significant ( P<0.05); the average BVN area, PED height, and SFCT decreased. The difference in average BVN area and PED height was statistically significant ( F=2.805, 3.916; P<0.05), and the difference in SFCT was not statistically significant ( F=0.047, P>0.05). Conclusions:The detection rate of PCV by OCTA is 86.36%. After PCV anti-vascular endothelial growth factor drug treatment, BVN area decrease and polyps subside. OCTA is an effective means for PCV diagnosis and follow-up after anti-VEGF drug treatment.

Objective:To retrieve, evaluate and summarize the clinical practice evidence of nursing care for critically ill patients with naso-intestinal tube feeding, so as provide a basis for correcting feeding intolerance and nutritional substandard.Methods:According to the "6S" evidence model, evidence on nursing care for critically ill patients with naso-intestinal tube feeding was retrieved in Registered Nurses' Association of Ontario, Cochrane Library, National Institute for Health and Clinical Excellence, Scottish Intercollegiate Guidelines Network, Guidelines International Network, Medive, PubMed, CINAHL, Embase, UpToDate, Joanna Briggs Institute Evidence-based Practice Database, China National Knowledge Infrastructure (CNKI) , WanFang Data, VIP database from January 31, 2015 to September 30, 2019. Evidence included guidelines, expert consensus, best practices, systematic reviews, evidence summary, and original research. Two researchers evaluated the quality of the included literature, extracted and summarized literature that met the quality standards.Results:A total of 24 articles were included, of which 2 guidelines were derived from the American Society of Critical Care Medicine and the American Society of Parenteral Enteral Nutrition, and the European Society of Parenteral Enteral Nutrition, 3 expert consensus, 1 systematic review, 4 evidence summaries, 10 randomized controlled studies, 3 quasi experiments, and 1 diagnostic test. This study summarized 24 evidences of clinical suitability, involving 10 aspects, namely, naso-intestinal tube placement indications, catheter material, placement position, placement method, confirmation method, feeding method, fixation, intolerance, catheter blockage and indwelling time limit.Conclusions:This study provides a management process and evidence-based evidence for critically ill patients with naso-intestinal tube feeding, which is conducive to promoting safe practice and in-depth research for the nursing staff.

Objective:To analyze the antibody level of phospholipase A2 receptor (PLA2R) in Chinese patients with primary membrane nephropathy (PMN) and its correlation with clinical indicators, and to explore more suitable cut-off value for Chinese patients.Methods:All hospitalized patients with renal biopsy at Peking University People's Hospital from January to August 2018 were retrospectively reviewed. According to the primary disease, patients were divided into PMN group (including patients with idiopathic membranous nephropathy and atypical membranous nephropathy of unknown cause) and control group (other pathological types, including secondary membranous nephropathy patients). Their clinical and pathological characteristics were analyzed, and the level of serum PLA2R antibodies was detected using the method of enzyme-linked immunosorbent assay (ELISA). Spearman correlation was used to analyze the correlation between PMN patients' blood anti-PLA2R antibody level and clinical indicators. The risk factors of PMN were analyzed by logistic regression model, and the optimal cut-off value of PMN was analyzed by ROC curve.Results:A total of 354 patients were included in this study, including 114 patients in PMN group and 240 patients in control group. The age of PMN group was (51.7±14.1) years old and the ratio of male to female was 2.2∶1. The median concentration of PLA2R antibody in PMN group was 16.87 RU/ml [inter-quartile range ( IQR) 1.88-57.26], which was significantly higher than that in control group (1.43 RU/ml, IQR 1.20-1.62, P<0.001). In PMN group, the concentration of anti-PLA2R antibody was correlated with the 24-hour urine protein ration ( r=0.278, P=0.003) and urine erythrocyte ( r=0.190, P=0.043), but not with serum albumin ( r=-0.149, P=0.114) and serum creatinine ( r=0.136, P=0.149). The ROC curve showed that the sensitivity of distinguishing PMN from other diseases was 69.3% (95% CI 60.3%-77.0%), the specificity was 92.9%(95% CI 89.0%-95.5%), and the area under the curve was 0.859(95% CI 0.813-0.905) when the cut-off value was set as 2.28 RU/ml, which was significantly better than the cut-off value of 20.00 RU/ml (the sensitivity/specificity was 46.5%/97.5%) and 14.00 RU/ml (the sensitivity/specificity was 53.5%/97.1%). Conclusions:PLA2R antibody is one of the main pathogenic antibodies of PMN. In China, it is recommended to lower its cut-off value to 2.28 RU/ml, which can improve the sensitivity of distinguishing PMN from other pathological types without reducing specificity.