Objective The aim of this study was to analyze the value of quantitative parameters of dynamic contrast en-hanced magnetic resonance imaging(DCE-MRI)combined with the detection of non-SMC condensing Ⅰ complex subunit H(NCAPH)in the diagnosis of early breast cancer.Methods Ninety-six patients with breast nodules who were treated in the depart-ment of Breast Surgery at Longgang District Maternity&Child Healthcare Hospital in Shenzhen from March 2020 to March 2022 were selected as the study objects.DCE-MRI examination was performed on all patients,and transport constant(Ktrans)and rate constant(Kep)were recorded.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of serum NCAPH mRNA.Based on the results of pathological examination as the gold standard,the patients with breast nodules diag-nosed pathologically as the benign group and the patients with breast cancer as the breast cancer group,the differences of DCE-MRI quantitative parameters Ktrans,Kep and serum NCAPH mRNA between the benign group and the breast cancer group were compared.The accuracy,sensitivity and specificity of serum NCAPH mRNA and their combination in the diagnosis of early breast cancer were different.Kappa test was used to compare the consistency with the pathological results.Results The results of pathological examina-tion confirmed that there were 31 benign nodules and 65 breast cancer in 96 patients with breast nodules.Ktrans,Kep and NCAPH mRNA in the breast cancer group were significantly higher than those in the benign group(P<0.05);The AUC of Ktrans,Kep and NCAPH in the diagnosis of early breast cancer was 0.944,which was significantly higher than that of Ktrans and Kep alone,with the sensitivity and specificity of 96.92% and 77.42% ,respectively;Ktrans and Kep combined with NCAPH detected 7 false positives and 2 false negatives,with a Kappa value of 0.776(P<0.05),which was consistent with the pathological results;The sensitivity of DCE-MRI quantitative parameters combined with NCAPH in the diagnosis of early breast cancer was significantly higher than that of DCE-MRI quantitative parameters and serum NCAPH alone(P<0.05).Conclusion The expression of Ktrans,Kep and serum NCAPH mRNA in breast cancer patients with DCE-MRI quantitative parameters is high.Ktrans,Kep combined with serum NCAPH detection has certain clinical values in the diagnosis of early breast cancer.

Objective:To investigate the role of Gasdermin E (GSDME) -mediated keratinocyte pyroptosis induced by Cutibacterium acnes ( C.acnes) in the pathogenesis of acne. Methods:The human immortalized keratinocyte HaCaT cells were stimulated with heat-inactived C.acnes for 15 minutes to 24 hours, and Western blot analysis was performed to determine the expression of cleaved GSDME (GSDME-NT) in HaCaT cells at different time points. Skin tissue samples were collected from 5 acne patients and 4 healthy controls, who visited the Hospital for Skin Diseases, Chinese Academy of Medical Sciences from January 2 to December 1, 2024; additionally, 3 samples of acne cyst contents and 3 samples of normal follicle contents were collected. Immunohistochemical study and Western blot analysis were conducted to determine GSDME-NT expression in the epidermis. Enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of interleukin (IL) -1β and tumor necrosis factor (TNF) -α in acne cyst or normal follicle contents. GSDME-knockdown HaCaT cells were constructed by transfection with lentivirus carrying GSDME-shRNA, and HaCaT cells transfected with lentivirus carrying the nonsense sequence control (NC) served as controls; ELISA was performed to detect the levels of IL-1β and TNF-α in GSDME-knockdown HaCaT cells after C. acnes stimulation ( C. acnes + GSDME knockdown group) , as well as in the phosphate-buffered saline (PBS) + NC group, C. acnes + NC group, and PBS + GSDME knockdown group. Western blot analysis was conducted to determine the GSDME-NT expression in HaCaT cells pretreated with or without retinol after C. acnes stimulation. Results:The cleavage of GSDME in HaCaT cells began at 1 hour after in vitro C. acnes stimulation, and GSDME-NT could be detected at this time. Compared with the control epidermis, the proportion of GSDME-NT-positive HaCaT cells (9.34% ± 2.92% vs. 3.05% ± 1.14%, t = -3.47, P = 0.026) and GSDME-NT protein expression levels ( t = -3.51, P = 0.025) significantly increased in the lesional epidermis of acne patients. The levels of IL-1β and TNF-α were significantly higher in the acne cyst contents than in the normal follicle contents (IL-1β: 1 337.24 [1 182.32, 2 230.61] pg/ml vs. 0.00 [0.00, 108.21] pg/ml, Z = 1.99, P = 0.046; TNF-α: 811.31 [438.26, 817.73] pg/ml vs. 46.67 [12.41, 53.21] pg/ml, Z = 1.96, P = 0.049) . ELISA showed that the levels of IL-1β and TNF-α were significantly higher in the C. acnes + NC group (12.12 ± 3.07 pg/ml, 26.06 ± 1.57 pg/ml, respectively) than in the PBS + NC group (3.73 ± 2.24 pg/ml, 10.14 ± 0.79 pg/ml, P = 0.003, < 0.001, respectively) ; compared with the C. acnes + NC group, the levels of IL-1β and TNF-α significantly decreased in the C. acnes + GSDME knockdown group (3.38 ± 0.93 pg/ml, 12.67 ± 2.10 pg/ml, P = 0.003, < 0.001, respectively) . The GSDME-NT expression was significantly lower in the retinol + C. acnes group than in the C. acnes group ( P = 0.029) . Conclusion:C. acnes may induce GSDME-mediated pyroptosis in keratinocytes, thereby promoting the release of inflammatory factors and aggravating the inflammatory response in acne, while retinol may be able to inhibit this process.

Objective:To investigate the current use of ulinastatin in the treatment of critically ill children by pediatricians in China.Methods:A anonymous questionnaire survey was conducted among 147 pediatric critical care physicians from 36 hospitals across 16 provinces,autonomous regions,and municipalities in China.The survey content consists of three parts: respondents' basic information, the application status of ulinastatin, and the clinical indicators referenced for evaluating the use of ulinastatin. Descriptive statistical analysis was performed on the collected data.Results:Among the 147 respondents,99.32%(146/147) were from tertiary hospitals；72.11%(106/147) worked in specialized ICUs,and 4.08%(6/147)in emergency medicine departments.A total of 68.03%(100/147) of the physicians reported using ulinastatin in clinical practice.The main diseases for which ulinastatin was used were pancreatitis(26.40%),sepsis and septic shock(23.76%),capillary leak syndrome(21.78%),acute respiratory distress syndrome(8.91%),and disseminated intravascular coagulation(6.27%).A total of 90.00% of physicians combined ulinastatin with other medications,including glucocorticoids(26.82%),albumin(23.51%),plasma(17.22%),and immunoglobulins(13.58%). Clinical indicators referenced during ulinastatin use included elevated interleukin(IL)-6(76.87%),tumor necrosis factor-α(44.22%),IL-8(31.97%),IL-1(19.73%),IL-18(10.20%),blood lactate(59.18%),decreased serum albumin levels(70.07%),increased pleural or peritoneal effusion(67.35%),skin and mucosal edema(65.31%),and elevated thrombomodulin among the four coagulation parameters(58.50%).Conclusion:Ulinastatin is mainly used for the treatment of critical illnesses such as pancreatitis and sepsis.Most physicians combine ulinastatin with other drugs,such as glucocorticoids and albumin.Clinical indicators commonly referenced when using ulinastatin include elevated IL-6,increased lactate,and increased pleural effusion,which suggest a high inflammatory state and endothelial damage.

Objective:To investigate the role of Gasdermin E (GSDME) -mediated keratinocyte pyroptosis induced by Cutibacterium acnes ( C.acnes) in the pathogenesis of acne. Methods:The human immortalized keratinocyte HaCaT cells were stimulated with heat-inactived C.acnes for 15 minutes to 24 hours, and Western blot analysis was performed to determine the expression of cleaved GSDME (GSDME-NT) in HaCaT cells at different time points. Skin tissue samples were collected from 5 acne patients and 4 healthy controls, who visited the Hospital for Skin Diseases, Chinese Academy of Medical Sciences from January 2 to December 1, 2024; additionally, 3 samples of acne cyst contents and 3 samples of normal follicle contents were collected. Immunohistochemical study and Western blot analysis were conducted to determine GSDME-NT expression in the epidermis. Enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of interleukin (IL) -1β and tumor necrosis factor (TNF) -α in acne cyst or normal follicle contents. GSDME-knockdown HaCaT cells were constructed by transfection with lentivirus carrying GSDME-shRNA, and HaCaT cells transfected with lentivirus carrying the nonsense sequence control (NC) served as controls; ELISA was performed to detect the levels of IL-1β and TNF-α in GSDME-knockdown HaCaT cells after C. acnes stimulation ( C. acnes + GSDME knockdown group) , as well as in the phosphate-buffered saline (PBS) + NC group, C. acnes + NC group, and PBS + GSDME knockdown group. Western blot analysis was conducted to determine the GSDME-NT expression in HaCaT cells pretreated with or without retinol after C. acnes stimulation. Results:The cleavage of GSDME in HaCaT cells began at 1 hour after in vitro C. acnes stimulation, and GSDME-NT could be detected at this time. Compared with the control epidermis, the proportion of GSDME-NT-positive HaCaT cells (9.34% ± 2.92% vs. 3.05% ± 1.14%, t = -3.47, P = 0.026) and GSDME-NT protein expression levels ( t = -3.51, P = 0.025) significantly increased in the lesional epidermis of acne patients. The levels of IL-1β and TNF-α were significantly higher in the acne cyst contents than in the normal follicle contents (IL-1β: 1 337.24 [1 182.32, 2 230.61] pg/ml vs. 0.00 [0.00, 108.21] pg/ml, Z = 1.99, P = 0.046; TNF-α: 811.31 [438.26, 817.73] pg/ml vs. 46.67 [12.41, 53.21] pg/ml, Z = 1.96, P = 0.049) . ELISA showed that the levels of IL-1β and TNF-α were significantly higher in the C. acnes + NC group (12.12 ± 3.07 pg/ml, 26.06 ± 1.57 pg/ml, respectively) than in the PBS + NC group (3.73 ± 2.24 pg/ml, 10.14 ± 0.79 pg/ml, P = 0.003, < 0.001, respectively) ; compared with the C. acnes + NC group, the levels of IL-1β and TNF-α significantly decreased in the C. acnes + GSDME knockdown group (3.38 ± 0.93 pg/ml, 12.67 ± 2.10 pg/ml, P = 0.003, < 0.001, respectively) . The GSDME-NT expression was significantly lower in the retinol + C. acnes group than in the C. acnes group ( P = 0.029) . Conclusion:C. acnes may induce GSDME-mediated pyroptosis in keratinocytes, thereby promoting the release of inflammatory factors and aggravating the inflammatory response in acne, while retinol may be able to inhibit this process.

Objective The aim of this study was to analyze the value of quantitative parameters of dynamic contrast en-hanced magnetic resonance imaging(DCE-MRI)combined with the detection of non-SMC condensing Ⅰ complex subunit H(NCAPH)in the diagnosis of early breast cancer.Methods Ninety-six patients with breast nodules who were treated in the depart-ment of Breast Surgery at Longgang District Maternity&Child Healthcare Hospital in Shenzhen from March 2020 to March 2022 were selected as the study objects.DCE-MRI examination was performed on all patients,and transport constant(Ktrans)and rate constant(Kep)were recorded.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of serum NCAPH mRNA.Based on the results of pathological examination as the gold standard,the patients with breast nodules diag-nosed pathologically as the benign group and the patients with breast cancer as the breast cancer group,the differences of DCE-MRI quantitative parameters Ktrans,Kep and serum NCAPH mRNA between the benign group and the breast cancer group were compared.The accuracy,sensitivity and specificity of serum NCAPH mRNA and their combination in the diagnosis of early breast cancer were different.Kappa test was used to compare the consistency with the pathological results.Results The results of pathological examina-tion confirmed that there were 31 benign nodules and 65 breast cancer in 96 patients with breast nodules.Ktrans,Kep and NCAPH mRNA in the breast cancer group were significantly higher than those in the benign group(P<0.05);The AUC of Ktrans,Kep and NCAPH in the diagnosis of early breast cancer was 0.944,which was significantly higher than that of Ktrans and Kep alone,with the sensitivity and specificity of 96.92% and 77.42% ,respectively;Ktrans and Kep combined with NCAPH detected 7 false positives and 2 false negatives,with a Kappa value of 0.776(P<0.05),which was consistent with the pathological results;The sensitivity of DCE-MRI quantitative parameters combined with NCAPH in the diagnosis of early breast cancer was significantly higher than that of DCE-MRI quantitative parameters and serum NCAPH alone(P<0.05).Conclusion The expression of Ktrans,Kep and serum NCAPH mRNA in breast cancer patients with DCE-MRI quantitative parameters is high.Ktrans,Kep combined with serum NCAPH detection has certain clinical values in the diagnosis of early breast cancer.

Objective:To investigate the current use of ulinastatin in the treatment of critically ill children by pediatricians in China.Methods:A anonymous questionnaire survey was conducted among 147 pediatric critical care physicians from 36 hospitals across 16 provinces,autonomous regions,and municipalities in China.The survey content consists of three parts: respondents' basic information, the application status of ulinastatin, and the clinical indicators referenced for evaluating the use of ulinastatin. Descriptive statistical analysis was performed on the collected data.Results:Among the 147 respondents,99.32%(146/147) were from tertiary hospitals；72.11%(106/147) worked in specialized ICUs,and 4.08%(6/147)in emergency medicine departments.A total of 68.03%(100/147) of the physicians reported using ulinastatin in clinical practice.The main diseases for which ulinastatin was used were pancreatitis(26.40%),sepsis and septic shock(23.76%),capillary leak syndrome(21.78%),acute respiratory distress syndrome(8.91%),and disseminated intravascular coagulation(6.27%).A total of 90.00% of physicians combined ulinastatin with other medications,including glucocorticoids(26.82%),albumin(23.51%),plasma(17.22%),and immunoglobulins(13.58%). Clinical indicators referenced during ulinastatin use included elevated interleukin(IL)-6(76.87%),tumor necrosis factor-α(44.22%),IL-8(31.97%),IL-1(19.73%),IL-18(10.20%),blood lactate(59.18%),decreased serum albumin levels(70.07%),increased pleural or peritoneal effusion(67.35%),skin and mucosal edema(65.31%),and elevated thrombomodulin among the four coagulation parameters(58.50%).Conclusion:Ulinastatin is mainly used for the treatment of critical illnesses such as pancreatitis and sepsis.Most physicians combine ulinastatin with other drugs,such as glucocorticoids and albumin.Clinical indicators commonly referenced when using ulinastatin include elevated IL-6,increased lactate,and increased pleural effusion,which suggest a high inflammatory state and endothelial damage.

Objective This project aimed to study the Miao medicine helleborus thibetanus franchon,including investigating its anti-inflammatory activity in collagen-induced arthritis CIA rats and its mechanism of VEGF/VEGFR2/P38 MAPK pathway regulation.Methods Sixty female Wistar rats were randomly divided into six groups:normal;model;positive drug;and low,medium,high dose groups,with 10 rats in each group.Bovine type Ⅱ collagen solution was injected into the tail of rats to construct the rheumatoid arthritis model,and the positive drug group was given MTX2.0 mg/(kg·d)by gavage once every other day.The three groups of helleborus thibetanus franchon low,medium,and high dose were gavaged with helleborus thibetanus franchon ethanol extract at 0.25,0.5 and 1 g/(kg·d)once a day.The normal and model groups were given an equivalent volume of NaCl solution,with continuous administration lasting for 28 days.During treatment,the general condition of the rats was observed,body weight changes recorded,and foot thickness measured.After treatment and euthanasia,the rats'hind limbs were removed for Micro-CT to detect bone destruction;hematoxylin and eosin staining for pathological investigattion of the synovial membrane;immunohistochemistry to observe neovascularization in the synovium;quantitative reverse-transcription PCR to detect mRNA levels of VEGF-A,VEGFR2,TNF-α in the synovial tissue;and Western Blot to detect the expression of VEGF,VEGFR2,p-P38,p-AKT.The analyses were used to explore the potential mechanisms of action of the Miao medicine helleborus thibetanus franchon in treating rheumatoid arthritis.Results Compared with the normal group,the model group showed significant weight loss(P＜0.01),increased foot swelling(P＜0.01),visible proliferative synovial tissue with inflammatory cell infiltration,erosive lesions on bone surfaces,increased neovascularization in the synovium,and significant bone destruction in Micro-CT,with reduced bone percentage,trabecular thickness,and bone density.The levels of VEGF-A,VEGFR2,TNF-α mRNA and VEGF-A,VEGFR2,p-P38,p-AKT proteins were significantly elevated(P＜0.01).Compared with the model group,the helleborus thibetanus franchon ethanol extract-treated groups showed improvements in these conditions in a dose-dependent manner,with the high-dose group receiving the best effect.There was a significant increase in the rats'body weight(P＜0.05);reduction in foot swelling(P＜0.05);amelioration of synovial and erosive bone lesions;reduction in neovascularization in the synovium;and significantly lower levels of VEGF-A,VEGFR2,and TNF-α mRNA,and VEGF-A,VEGFR2,p-P38,and p-AKT protein(P＜0.01).Conclusions The Miao medicine plant helleborus thibetanus franchon may alleviate joint inflammatory damage in CIA rats by modulating the VEGF/VEGFR2 signaling pathway,thereby exerting therapeutic effects on rheumatoid arthritis.

Objective To investigate the regulatory role of miR-26b-3p in proliferation,migration and invasion of glioma.Methods The expressions of miR-26b-3p and cAMP-responsive element binding protein 1(CREB1)in gliomas of different pathological grades were detected with RT-qPCR and Western blotting.Bioinformatic methods were used to analyze the target sequence of miRNA-26b-3p binding to CREB1,and dual luciferase gene reporter experiment was performed to explore the mechanism for targeted regulation of CREB1 by miR-26b-3p.Glioma U251 cells were treated with miR-26b-3p mimic or inhibitor,and the changes in CREB1 expression and cell proliferation,migration,invasion and apoptosis were determined with Western blotting,CCK-8 assay,wound healing assay,Transwell assay,and flow cytometry.Results The expression of miR-26b-3p decreased while CREB1 expression increased significantly as the pathological grade of gliomas increased(P<0.05).Dual luciferase gene reporter experiment confirmed that CREB1 was a downstream target of miR-26b-3p.Inhibition of miR-26b-3p significantly upregulated the expression of CERB1,suppressed apoptosis and promoted proliferation and invasion of glioma cells,and overexpression of miR-26b-3p produced the opposite effects(P<0.05).Conclusion MiR-26b-3p regulates CREB1 expression to modulate apoptosis,proliferation,migration and invasion of glioma cells,thereby participating in tumorigenesis and progression of glioma.

Objective To investigate the regulatory role of miR-26b-3p in proliferation,migration and invasion of glioma.Methods The expressions of miR-26b-3p and cAMP-responsive element binding protein 1(CREB1)in gliomas of different pathological grades were detected with RT-qPCR and Western blotting.Bioinformatic methods were used to analyze the target sequence of miRNA-26b-3p binding to CREB1,and dual luciferase gene reporter experiment was performed to explore the mechanism for targeted regulation of CREB1 by miR-26b-3p.Glioma U251 cells were treated with miR-26b-3p mimic or inhibitor,and the changes in CREB1 expression and cell proliferation,migration,invasion and apoptosis were determined with Western blotting,CCK-8 assay,wound healing assay,Transwell assay,and flow cytometry.Results The expression of miR-26b-3p decreased while CREB1 expression increased significantly as the pathological grade of gliomas increased(P<0.05).Dual luciferase gene reporter experiment confirmed that CREB1 was a downstream target of miR-26b-3p.Inhibition of miR-26b-3p significantly upregulated the expression of CERB1,suppressed apoptosis and promoted proliferation and invasion of glioma cells,and overexpression of miR-26b-3p produced the opposite effects(P<0.05).Conclusion MiR-26b-3p regulates CREB1 expression to modulate apoptosis,proliferation,migration and invasion of glioma cells,thereby participating in tumorigenesis and progression of glioma.

Lipophagy, the selective engulfment of lipid droplets (LDs) by autophagosomes for lysosomal degradation, is critical to lipid and energy homeostasis. Here we show that the lipid transfer protein ORP8 is located on LDs and mediates the encapsulation of LDs by autophagosomal membranes. This function of ORP8 is independent of its lipid transporter activity and is achieved through direct interaction with phagophore-anchored LC3/GABARAPs. Upon lipophagy induction, ORP8 has increased localization on LDs and is phosphorylated by AMPK, thereby enhancing its affinity for LC3/GABARAPs. Deletion of ORP8 or interruption of ORP8-LC3/GABARAP interaction results in accumulation of LDs and increased intracellular triglyceride. Overexpression of ORP8 alleviates LD and triglyceride deposition in the liver of ob/ob mice, and Osbpl8-/- mice exhibit liver lipid clearance defects. Our results suggest that ORP8 is a lipophagy receptor that plays a key role in cellular lipid metabolism.
Animals ; Mice ; Lipid Droplets ; Autophagy ; Autophagosomes ; Homeostasis ; Triglycerides