Objective:To explore the applicability of modified comprehensive interventions in the treatment of non-severe dry eye syndrome in military pilots.Methods:A total of 88 military pilots with non-severe dry eye syndrome admitted to the Special Service Department of the 988th Hospital of the Joint Logistic Support Force between December 2021 and December 2023 were divided into an intervention group and a control group using the random number table method, with 44 cases in each. The intervention group received modified comprehensive interventions, while the control group underwent conventional treatment. The Ocular Surface Disease Index (OSDI), break-up time, tear meniscus height, changes in meibomian gland function, and levels of satisfaction of military pilots were compared between the 2 groups. The correlations between the OSDI, break-up time, tear meniscus height and levels of satisfaction were analyzed.Results:Before treatment, there was no significant difference in the OSDI between the 2 groups ( P>0.05). After 4 weeks of treatment, the changes in the OSDI of military pilots were smaller in the intervention group than in the control group ( t=3.21, P=0.002). After 2 and 4 weeks of treatment, the break-up time (both P<0.001) and tear meniscus height ( P<0.001, =0.012) of pilots in the intervention group exceeded those of the control group. In both groups, the break-up time (all P<0.001) and tear meniscus height (all P<0.001) kept increasing after treatment. After 4 weeks of treatment, there were significant differences in the distribution of meibomian gland function between the 2 groups ( Z=-2.55, -2.41, -2.29, P=0.011, 0.016, 0.022). Clinical care, procedure flow, and health education scored higher in the intervention group than in the control group during the survey on levels of satisfaction with the treatment ( t=6.55, 6.77, 3.63, all P≤0.001). The OSDI was negatively correlated with clinical care, procedure flow and health education ( r=-0.286, -0.275, -0.363, P=0.007, 0.010, 0.001) while the break-up time was positively correlated with clinical care and procedure flow ( r=0.248, 0.278, P=0.020, 0.009). Conclusions:The implementation of modified comprehensive intervention measures for dry eye syndrome in military pilots can effectively improve clinical symptoms and leave military pilots more satisfied.

Objective To explore the role of nucleolin 6(NOL6)in the occurrence and development of hypertension and its mechanism of regulating ribosome biogenesis.Methods Differentially expressed genes were screened based on the GEO database(chip GSE212338),and intersection analysis was conducted in combination with genes related to ribosome generation to obtain genes related to ribosome biogenesis in hypertension.The rats were divided into control group and model group(L-NAME group).The hypertensive rat model was induced by N-nitro-L-arginine methyl ester(L-NAME),and the thickness and pathological changes of the aortic wall in each group were observed by HE staining.The expression of ribosomal RNA(rRNA)in rat aortic tissues was detected by qPCR to reflect ribosome biogenesis,and the protein expression of NOL6 was detected by Western blotting.Human umbilical vein endothelial cells(HUVECs)were cultured and grouped for treatment(control group,L-NAME group,AngⅡ group,AngⅡ+si-NC group,AngⅡ+si-NOL6 group,and AngⅡ+CX-5461 group).The generation of neocRNA in HUVEC was detected by EU.The protein and mRNA expressions of NOL6 in HUVEC were detected by Western blotting and qPCR,respectively.Western blotting was used to detect the protein expressions of endothelial nitric oxide synthase(eNOS)and p-eNOS.Results By combining the differential expression analysis of the GEO hypertension dataset GSE212338 and the ribosome biogenesis gene set,six core genes with significantly altered expression in hypertension and related to ribosome biogenesis were identified.The difference in NOL6 was the most significant.Compared with the control group,the aortic wall thickness of rats in the L-NAME group increased significantly.Ribosomal RNA expression was significantly upregulated;the protein and mRNA expressions of NOL6 were significantly upregulated,too.Compared with the control group,the generation of neoRNA in the cells of the L-NAME group increased significantly;the levels of NOL6 protein and mRNA,ribosomal RNA and neoRNA in the Ang Ⅱ group were significantly increased compared with the control group but significantly decreased compared with the Ang Ⅱ+si-NC group.Compared with the Ang Ⅱ+si-NOL6 group,the protein and mRNA expressions of NOL6 in the AngⅡ+si-NC group and the AngⅡ+CX-5461 group cells were significantly increased.Compared with the AngⅡ+si-NC group,the levels of ribosomal RNA and neoRNA in the AngⅡ+si-NOL6 group and the AngⅡ+CX-5461 group were significantly decreased;the protein expressions of eNOS and p-eNOS were significantly increased.Conclusion NOL6 is associated with abnormal ribosome biogenesis in hypertension.NOL6 can affect the expression of eNOS by regulating ribosome biogenesis,thereby regulating the occurrence and development of hypertension.

Objective:To analyze the efficacy and prognostic factors of fractionated stereotactic radiotherapy (FSRT) for patients with breast cancer brain metastases (BCBM).Methods:Medical records and follow-up data of BCBM patients who underwent FSRT in Cancer Hospital Chinese Academy of Medical Sciences, Shenzhen Center and Shenzhen People's Hospital from August 2019 to May 2023 were collected. The R Studio platform of the R version 4.2.1 statistical software was applied to analyze patients' baseline characteristics, 1- and 2-year local brain control (LBC), overall survival (OS) and distant brain control (DBC) and corresponding median failure-free survival, draw survival curve using Kaplan-Meier method. Prognostic factors were screened by univariate analysis and multivariate analysis (Cox regression).Results:Cumulatively, 52 patients (163 metastases in total) had a median survival follow-up of 22.1 months, 83% were<60 years old. Molecular typing: 13 cases (25%) were positive for human epidermal growth factor receptor 2 (HER2+) / hormone receptor negative (HR-), 2 cases (4%) were luminal A, 26 cases (50%) were luminal B, and 11 cases (21%) were triple negative. The median number of brain metastases was 2 (range: 1 - 17). Follow-up outcomes: the median OS was 34.0 months, with 1- and 2-year OS rates of 85.6% and 65.4%, respectively; the median LBC was 20.6 months, with 1- and 2-year LBC rates of 79.2% and 45.2%, respectively; and the median DBC was 10.3 months, with 1- and 2-year DBC rates of 46.7% and 28.9%, respectively. During follow-up, 13 patients underwent salvage local therapy (10 FSRT); 5 developed radiation necrosis (1 symptomatic). Prognostic factor analysis: absence of extracranial organ metastases (compared with ≥3) was a protective factor for OS, P<0.05. For LBC, fewer (1 - 2) extracranial organ metastases (compared with ≥3), and single brain metastasis (compared with ≥2) were favorable prognostic factors , while N 3 staging upon initial diagnosis was a poor prognostic factor (all P<0.05). For DBC, brain metastasis after surgery was a good prognostic factor, while complicated with lung metastasis and asymptomatic brain metastasis at the first diagnosis were poor prognostic factors (all P<0.05). Conclusions:FSRT yields relatively good LBC and poor DBC for BCBM patients. A certain percentage of patients require salvage FSRT during follow-up, but OS is maintained acceptable and the radiation necrosis is tolerable. Among the prognostic factors, the absence of extracranial metastatic organs is a good prognostic factor for OS; patients with single brain metastasis, fewer extracranial metastatic organs, and non-N 3 staging upon initial diagnosis can obtain better LBC after FSRT.

Objective:To investigate whether brown adipose tissue-derived exosomes(BAT-exos) could ameliorate endothelial cell injury by activating Pink1-Parkin pathway-mediated mitophagy.Methods:Endothelial cell injury was induced with angiotensin Ⅱ(Ang Ⅱ) to establish a cellular injury model. Exosomes were isolated from both brown adipose tissue and white adipose tissue and characterized by transmission electron microscopy(TEM), nanoparticle tracking analysis(NTA), fluorescence labeling, and Western blot. Cell viability was assessed using the CCK-8 assay, and apoptosis rates were determined by flow cytometry. Levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-8 were measured by ELISA. Mitochondrial autophagy was assessed by immunofluorescence colocalization, and protein expression levels of Pink1, Parkin, and LC3 Ⅱ/I were determined by Western blot.Results:Ang Ⅱ induced endothelial cell apoptosis, activated inflammatory responses, and suppressed mitophagy, as evidenced by decreased expression of mitophagy-related proteins. Following the successful characterization of BAT-exos, we found that BAT-exos activated mitophagy and alleviated endothelial cell injury, whereas white adipose tissue-derived exosomes(WAT-exos) inhibited mitophagy and exacerbated injury. Mechanistically, BAT-exos targeted the Pink1-Parkin signaling pathway to activate mitophagy.Conclusion:BAT-exos markedly improve endothelial cell injury by activating mitophagy through the Pink1-Parkin pathway, providing new insights and potential therapeutic targets for cardiovascular diseases.

AIM:To explore the mechanism of Salvia miltiorrhiza(Danshen)-derived exosome-like nanoparti-cles(DDN)in attenuating oxidative damage in endothelial cells through the activation of the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/Akt)/endothelial nitric oxide synthase(eNOS)signaling pathway.METHODS:The DDN were characterized by transmission electron microscopy and dynamic light scattering.Fluorescence microscopy and flow cytometry were used to evaluate the uptake of DDN by human umbilical vein endothelial cells(HUVECs).The viability,migration and invasion of HUVECs were assessed using CCK8 assay,wound-healing assay and Transwell assay,respec-tively.The HUVECs were induced by angiotensin II(Ang II)for oxidative stress and intervened with DDN or LY294002(a PI3K inhibitor).The levels of reactive oxygen species were determined by flow cytometry,and intracellular nitric oxide(NO)content was measured using a biochemical assay kit.Additionally,the protein levels of NADPH oxidase 4(NOX4),NOX2,endothelial nitric oxide syntnase(eNOS),p-eNOS,Akt and p-Akt were examined by Western blot.RESULTS:(1)Transmission electron microscopy and dynamic light scattering analysis revealed that DDN had good bio-compatibility and stability.(2)According to fluorescence images and flow cytometry results,DDN were strongly taken up by HUVECs.(3)Compared with control group,DDN significantly promoted the viability,migration and invasion of HUVECs,showing a dose-dependent effect.(4)Compared with control group,DDN remarkably increased intracellular NO levels,thereby enhancing endothelial cell vasodilation via activating the PI3K/Akt/eNOS signaling pathway.(5)The PI3K/Akt/eNOS pathway played a critical role in mitigating oxidative stress and improving cellular function in response to DDN treat-ment.CONCLUSION:The DDN mediate PI3K/Akt/eNOS signaling pathway activation to significantly alleviate Ang II-induced oxidative damage in endothelial cells,suggesting a potential vascular protective effect of DDN.

AIM:To explore the mechanism of Salvia miltiorrhiza(Danshen)-derived exosome-like nanoparti-cles(DDN)in attenuating oxidative damage in endothelial cells through the activation of the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/Akt)/endothelial nitric oxide synthase(eNOS)signaling pathway.METHODS:The DDN were characterized by transmission electron microscopy and dynamic light scattering.Fluorescence microscopy and flow cytometry were used to evaluate the uptake of DDN by human umbilical vein endothelial cells(HUVECs).The viability,migration and invasion of HUVECs were assessed using CCK8 assay,wound-healing assay and Transwell assay,respec-tively.The HUVECs were induced by angiotensin II(Ang II)for oxidative stress and intervened with DDN or LY294002(a PI3K inhibitor).The levels of reactive oxygen species were determined by flow cytometry,and intracellular nitric oxide(NO)content was measured using a biochemical assay kit.Additionally,the protein levels of NADPH oxidase 4(NOX4),NOX2,endothelial nitric oxide syntnase(eNOS),p-eNOS,Akt and p-Akt were examined by Western blot.RESULTS:(1)Transmission electron microscopy and dynamic light scattering analysis revealed that DDN had good bio-compatibility and stability.(2)According to fluorescence images and flow cytometry results,DDN were strongly taken up by HUVECs.(3)Compared with control group,DDN significantly promoted the viability,migration and invasion of HUVECs,showing a dose-dependent effect.(4)Compared with control group,DDN remarkably increased intracellular NO levels,thereby enhancing endothelial cell vasodilation via activating the PI3K/Akt/eNOS signaling pathway.(5)The PI3K/Akt/eNOS pathway played a critical role in mitigating oxidative stress and improving cellular function in response to DDN treat-ment.CONCLUSION:The DDN mediate PI3K/Akt/eNOS signaling pathway activation to significantly alleviate Ang II-induced oxidative damage in endothelial cells,suggesting a potential vascular protective effect of DDN.

Objective To explore the role of nucleolin 6(NOL6)in the occurrence and development of hypertension and its mechanism of regulating ribosome biogenesis.Methods Differentially expressed genes were screened based on the GEO database(chip GSE212338),and intersection analysis was conducted in combination with genes related to ribosome generation to obtain genes related to ribosome biogenesis in hypertension.The rats were divided into control group and model group(L-NAME group).The hypertensive rat model was induced by N-nitro-L-arginine methyl ester(L-NAME),and the thickness and pathological changes of the aortic wall in each group were observed by HE staining.The expression of ribosomal RNA(rRNA)in rat aortic tissues was detected by qPCR to reflect ribosome biogenesis,and the protein expression of NOL6 was detected by Western blotting.Human umbilical vein endothelial cells(HUVECs)were cultured and grouped for treatment(control group,L-NAME group,AngⅡ group,AngⅡ+si-NC group,AngⅡ+si-NOL6 group,and AngⅡ+CX-5461 group).The generation of neocRNA in HUVEC was detected by EU.The protein and mRNA expressions of NOL6 in HUVEC were detected by Western blotting and qPCR,respectively.Western blotting was used to detect the protein expressions of endothelial nitric oxide synthase(eNOS)and p-eNOS.Results By combining the differential expression analysis of the GEO hypertension dataset GSE212338 and the ribosome biogenesis gene set,six core genes with significantly altered expression in hypertension and related to ribosome biogenesis were identified.The difference in NOL6 was the most significant.Compared with the control group,the aortic wall thickness of rats in the L-NAME group increased significantly.Ribosomal RNA expression was significantly upregulated;the protein and mRNA expressions of NOL6 were significantly upregulated,too.Compared with the control group,the generation of neoRNA in the cells of the L-NAME group increased significantly;the levels of NOL6 protein and mRNA,ribosomal RNA and neoRNA in the Ang Ⅱ group were significantly increased compared with the control group but significantly decreased compared with the Ang Ⅱ+si-NC group.Compared with the Ang Ⅱ+si-NOL6 group,the protein and mRNA expressions of NOL6 in the AngⅡ+si-NC group and the AngⅡ+CX-5461 group cells were significantly increased.Compared with the AngⅡ+si-NC group,the levels of ribosomal RNA and neoRNA in the AngⅡ+si-NOL6 group and the AngⅡ+CX-5461 group were significantly decreased;the protein expressions of eNOS and p-eNOS were significantly increased.Conclusion NOL6 is associated with abnormal ribosome biogenesis in hypertension.NOL6 can affect the expression of eNOS by regulating ribosome biogenesis,thereby regulating the occurrence and development of hypertension.

Objective:To investigate whether brown adipose tissue-derived exosomes(BAT-exos) could ameliorate endothelial cell injury by activating Pink1-Parkin pathway-mediated mitophagy.Methods:Endothelial cell injury was induced with angiotensin Ⅱ(Ang Ⅱ) to establish a cellular injury model. Exosomes were isolated from both brown adipose tissue and white adipose tissue and characterized by transmission electron microscopy(TEM), nanoparticle tracking analysis(NTA), fluorescence labeling, and Western blot. Cell viability was assessed using the CCK-8 assay, and apoptosis rates were determined by flow cytometry. Levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-8 were measured by ELISA. Mitochondrial autophagy was assessed by immunofluorescence colocalization, and protein expression levels of Pink1, Parkin, and LC3 Ⅱ/I were determined by Western blot.Results:Ang Ⅱ induced endothelial cell apoptosis, activated inflammatory responses, and suppressed mitophagy, as evidenced by decreased expression of mitophagy-related proteins. Following the successful characterization of BAT-exos, we found that BAT-exos activated mitophagy and alleviated endothelial cell injury, whereas white adipose tissue-derived exosomes(WAT-exos) inhibited mitophagy and exacerbated injury. Mechanistically, BAT-exos targeted the Pink1-Parkin signaling pathway to activate mitophagy.Conclusion:BAT-exos markedly improve endothelial cell injury by activating mitophagy through the Pink1-Parkin pathway, providing new insights and potential therapeutic targets for cardiovascular diseases.

Objective:To explore the applicability of modified comprehensive interventions in the treatment of non-severe dry eye syndrome in military pilots.Methods:A total of 88 military pilots with non-severe dry eye syndrome admitted to the Special Service Department of the 988th Hospital of the Joint Logistic Support Force between December 2021 and December 2023 were divided into an intervention group and a control group using the random number table method, with 44 cases in each. The intervention group received modified comprehensive interventions, while the control group underwent conventional treatment. The Ocular Surface Disease Index (OSDI), break-up time, tear meniscus height, changes in meibomian gland function, and levels of satisfaction of military pilots were compared between the 2 groups. The correlations between the OSDI, break-up time, tear meniscus height and levels of satisfaction were analyzed.Results:Before treatment, there was no significant difference in the OSDI between the 2 groups ( P>0.05). After 4 weeks of treatment, the changes in the OSDI of military pilots were smaller in the intervention group than in the control group ( t=3.21, P=0.002). After 2 and 4 weeks of treatment, the break-up time (both P<0.001) and tear meniscus height ( P<0.001, =0.012) of pilots in the intervention group exceeded those of the control group. In both groups, the break-up time (all P<0.001) and tear meniscus height (all P<0.001) kept increasing after treatment. After 4 weeks of treatment, there were significant differences in the distribution of meibomian gland function between the 2 groups ( Z=-2.55, -2.41, -2.29, P=0.011, 0.016, 0.022). Clinical care, procedure flow, and health education scored higher in the intervention group than in the control group during the survey on levels of satisfaction with the treatment ( t=6.55, 6.77, 3.63, all P≤0.001). The OSDI was negatively correlated with clinical care, procedure flow and health education ( r=-0.286, -0.275, -0.363, P=0.007, 0.010, 0.001) while the break-up time was positively correlated with clinical care and procedure flow ( r=0.248, 0.278, P=0.020, 0.009). Conclusions:The implementation of modified comprehensive intervention measures for dry eye syndrome in military pilots can effectively improve clinical symptoms and leave military pilots more satisfied.

Objective:To analyze the efficacy and prognostic factors of fractionated stereotactic radiotherapy (FSRT) for patients with breast cancer brain metastases (BCBM).Methods:Medical records and follow-up data of BCBM patients who underwent FSRT in Cancer Hospital Chinese Academy of Medical Sciences, Shenzhen Center and Shenzhen People's Hospital from August 2019 to May 2023 were collected. The R Studio platform of the R version 4.2.1 statistical software was applied to analyze patients' baseline characteristics, 1- and 2-year local brain control (LBC), overall survival (OS) and distant brain control (DBC) and corresponding median failure-free survival, draw survival curve using Kaplan-Meier method. Prognostic factors were screened by univariate analysis and multivariate analysis (Cox regression).Results:Cumulatively, 52 patients (163 metastases in total) had a median survival follow-up of 22.1 months, 83% were<60 years old. Molecular typing: 13 cases (25%) were positive for human epidermal growth factor receptor 2 (HER2+) / hormone receptor negative (HR-), 2 cases (4%) were luminal A, 26 cases (50%) were luminal B, and 11 cases (21%) were triple negative. The median number of brain metastases was 2 (range: 1 - 17). Follow-up outcomes: the median OS was 34.0 months, with 1- and 2-year OS rates of 85.6% and 65.4%, respectively; the median LBC was 20.6 months, with 1- and 2-year LBC rates of 79.2% and 45.2%, respectively; and the median DBC was 10.3 months, with 1- and 2-year DBC rates of 46.7% and 28.9%, respectively. During follow-up, 13 patients underwent salvage local therapy (10 FSRT); 5 developed radiation necrosis (1 symptomatic). Prognostic factor analysis: absence of extracranial organ metastases (compared with ≥3) was a protective factor for OS, P<0.05. For LBC, fewer (1 - 2) extracranial organ metastases (compared with ≥3), and single brain metastasis (compared with ≥2) were favorable prognostic factors , while N 3 staging upon initial diagnosis was a poor prognostic factor (all P<0.05). For DBC, brain metastasis after surgery was a good prognostic factor, while complicated with lung metastasis and asymptomatic brain metastasis at the first diagnosis were poor prognostic factors (all P<0.05). Conclusions:FSRT yields relatively good LBC and poor DBC for BCBM patients. A certain percentage of patients require salvage FSRT during follow-up, but OS is maintained acceptable and the radiation necrosis is tolerable. Among the prognostic factors, the absence of extracranial metastatic organs is a good prognostic factor for OS; patients with single brain metastasis, fewer extracranial metastatic organs, and non-N 3 staging upon initial diagnosis can obtain better LBC after FSRT.