Objective To evaluate the sensitivity,repeatability,anti-degradation ability,in vivo temporal stability,and tissue specificity of the DNA methylation sequencing panel constructed in our previous study for height inference,so as to provide a reference for forensic application.Methods Sensitivity:different initial template quantities(50 ng,40 ng,30 ng,20 ng)were set for sequencing.Repeatability:DNA from the same sample was sequenced three times.Anti-degradation ability:whole blood was used to make blood stains,and DNA was extracted and sequenced at 0,3,6 and 9 months,respectively.In vivo temporal stability:the blood was collected at 0,3,6,and 9 months for sequencing.Tissue specificities:published data and findings were used to analyze the tissue specificities of CpGs in the panel.Results The sensitivity test showed that the initial template quantities of 20 ng detected all the CpG sites and still obtained accurate prediction results.The results of the three repeated predictions of the same sample are stable,and the differences are mainly due to the randomness of the DNN model,indicating good detection repeatability.A complete methylation profile was obtained for the blood stains left at room temperature for 9 months,and the predicted results showed a small range of fluctuations.The three samples were predicted to fluctuate within a range of 1.5 cm or less over nine months.Tissue-specific analyses showed a high correlation between blood and saliva,but can not apply to other tissues.Conclusion The DNA methylation detection system we developed in our previous study has good sensitivity,repeatability,anti degradation ability,in vivo time stability,as well as strong tissue specificity,making it suitable for height inference of blood samples.This supports the feasibility of using targeted DNA methylation analysis on whole blood samples to infer height in the field of forensic science.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

Objective This study aimed to investigate the tumorigenic properties of MMTV-PyMT breast cancer transgenic mice at different ages(in weeks)and the changes in the composition of immune cells in the tumor microenvironment.Methods Eight groups of 4,6,8,10,12,14,16 and 18 weeks of age MMTV-PyMT female mice(FVB mice as the background)and one group of 8 weeks of FVB female mice were prepared for routine blood testing,the pathological changes of the mammary gland and lung metastases were observed by histopathological sections,and the immune cells in blood,spleen,and tumor were analyzed by flow cytometry.Results MMTV-PyMT mice showed adenular ductal lesions at 4～6 weeks of age;the ductal portion expanded to the growth boundary at 8～9 weeks of age,and then gradually broke through the glandular boundary to form early breast cancer at 8～12 weeks of age,and advanced breast cancer at 10～14 weeks of age.At 12 weeks of age,metastases were visible in the lungs of some mice,and at 14 weeks of age,the number of metastases in the lungs increased significantly.As the age of the mice increased,the number of white blood cells,neutrophils,and platelets in their blood increased gradually,while the lymphocytes and erythrocytes showed a gradual downward trend.Flow cytometry showed that with the increase in age,the proportion of T cells in the spleen and tumor gradually decreased,the MDSCs in the blood,spleen,and tumor gradually increased,and the NK cells in the tumor also gradually increased.Conclusions This study analyzed routine blood tests,pathology,and immune cells in the tissues of MMTV-PyMT mouse models of different weeks of age,providing a novel perspective on the dynamic alterations of the tumor immune microenvironment during the malignant progression of breast cancer.

Objective By using ultrasonography to calculate the ratio of optic nerve sheath diameter(ONSD)at 3 mm behind eyeball to the eyeball transverse diameter(ETD),based on which to evaluate the effect of dexmedetomidine on intracranial pressure(ICP)in patients with intracranial aneurysm after receiving interventional embolization under general anesthesia.Methods A total of 40 patients with intracranial aneurysm,who were scheduled to receive interventional embolization under general anesthesia at the Affiliated Lihuili Hospital of Ningbo University of China from May 2023 to November 2023,were selected for this study.By using random number table method,the patients were divided into control group(group C)and dexmedetomidine group(group D)with 20 patients in each group.Standardized anesthesia strategy was adopted in both groups.For patients of group D,a loading dose of dexmedetomidine was pumped at a velocity of 1 μg/(kg·h)for 10 min before the surgery,the pumping was continued at 0.5 μg/(kg·h)velocity during the operation,and the pumping stopped half an hour before the end of the operation.For patients of group C,the same anesthesia strategy was used,while no any special treatment was given.At the different time points,including before awakening(T0),immediately after extubation(T1),and 5 min(T2),10 min(T3),15 min(T4)after extubation,ONSD at 3 mm behind eyeball and ETD were measured by transorbital ultrasonography,and ONSD/ETD ratio was calculated to evaluate ICP.The mean arterial pressure(MAP),heart rate(HR),blood oxygen saturation(SPO2),severity of cough,and extubation time were recorded at the time to remove the catheter.Results Compared with the data obtained at T0,the ONSD/ETD ratios obtained at T1 and T2 were increased in both groups(P＜0.05),while the ONSD/ETD ratios obtained at other time points were not significantly different from the ONSD/ETD ratio obtained at T0(P＞0.05).Compared with Group C,in Group D the ONSD/ETD ratios obtained at T1 and T2 were smaller,the differences were statistically significant(P＜0.05).The incidence of moderate to severe cough in Group D was lower than that in Group C(P＜0.05).Compared with Group C,in Group D the HR and MAP determined at the time of removing catheter were lower(P＜0.05).The extubation time in Group D was longer than that in group C,the difference was statistically significant(P＜0.05).Conclusion ONSD/ETD ratio calculated by ultrasono-graphy can objectively reflect the changes of ICP during the extubation period.Dexmedetomidine can reduce the elevation degree of ICP through effectively inhibiting cough reflex and circulatory fluctuation during tracheal extubation.However,dexmedetomidine may increase the incidence of adverse events such as bradycardia,delayed extubation,etc.

Objective This study aimed to investigate the tumorigenic properties of MMTV-PyMT breast cancer transgenic mice at different ages(in weeks)and the changes in the composition of immune cells in the tumor microenvironment.Methods Eight groups of 4,6,8,10,12,14,16 and 18 weeks of age MMTV-PyMT female mice(FVB mice as the background)and one group of 8 weeks of FVB female mice were prepared for routine blood testing,the pathological changes of the mammary gland and lung metastases were observed by histopathological sections,and the immune cells in blood,spleen,and tumor were analyzed by flow cytometry.Results MMTV-PyMT mice showed adenular ductal lesions at 4～6 weeks of age;the ductal portion expanded to the growth boundary at 8～9 weeks of age,and then gradually broke through the glandular boundary to form early breast cancer at 8～12 weeks of age,and advanced breast cancer at 10～14 weeks of age.At 12 weeks of age,metastases were visible in the lungs of some mice,and at 14 weeks of age,the number of metastases in the lungs increased significantly.As the age of the mice increased,the number of white blood cells,neutrophils,and platelets in their blood increased gradually,while the lymphocytes and erythrocytes showed a gradual downward trend.Flow cytometry showed that with the increase in age,the proportion of T cells in the spleen and tumor gradually decreased,the MDSCs in the blood,spleen,and tumor gradually increased,and the NK cells in the tumor also gradually increased.Conclusions This study analyzed routine blood tests,pathology,and immune cells in the tissues of MMTV-PyMT mouse models of different weeks of age,providing a novel perspective on the dynamic alterations of the tumor immune microenvironment during the malignant progression of breast cancer.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

Objective To evaluate the sensitivity,repeatability,anti-degradation ability,in vivo temporal stability,and tissue specificity of the DNA methylation sequencing panel constructed in our previous study for height inference,so as to provide a reference for forensic application.Methods Sensitivity:different initial template quantities(50 ng,40 ng,30 ng,20 ng)were set for sequencing.Repeatability:DNA from the same sample was sequenced three times.Anti-degradation ability:whole blood was used to make blood stains,and DNA was extracted and sequenced at 0,3,6 and 9 months,respectively.In vivo temporal stability:the blood was collected at 0,3,6,and 9 months for sequencing.Tissue specificities:published data and findings were used to analyze the tissue specificities of CpGs in the panel.Results The sensitivity test showed that the initial template quantities of 20 ng detected all the CpG sites and still obtained accurate prediction results.The results of the three repeated predictions of the same sample are stable,and the differences are mainly due to the randomness of the DNN model,indicating good detection repeatability.A complete methylation profile was obtained for the blood stains left at room temperature for 9 months,and the predicted results showed a small range of fluctuations.The three samples were predicted to fluctuate within a range of 1.5 cm or less over nine months.Tissue-specific analyses showed a high correlation between blood and saliva,but can not apply to other tissues.Conclusion The DNA methylation detection system we developed in our previous study has good sensitivity,repeatability,anti degradation ability,in vivo time stability,as well as strong tissue specificity,making it suitable for height inference of blood samples.This supports the feasibility of using targeted DNA methylation analysis on whole blood samples to infer height in the field of forensic science.

Objective:To investigate the clinical, radiological, and pathological features of anaplastic gangliogliomas (AGGs) and to determine whether these tumors represent a distinct entity.Methods:Consecutive 667 cases of ganglioglioma (GG) diagnosed at the Xuanwu Hospital, Capital Medical University, Beijing, China between January 2015 and July 2023 were screened. Among these cases, 9 pathologically confirmed AGG cases were identified. Their clinical, radiological, treatment, and outcome data were analyzed retrospectively. Most of the tumor samples were subject to next-generation sequencing, while a subset of them were subject to DNA methylation profiling.Results:Among the 9 patients, there were five males and four females, with a median age of 8 years. Epileptic seizures (5/9) were the most frequently presented symptom. Radiological examinations showed three types of radiological manifestations: four cases showed abnormal MRI signals with no significant mass effects and mild enhancement; two cases demonstrated a mixed solid-cystic density lesion with peritumoral edema, which showed significant heterogeneous enhancement and obvious mass effects, and one case displayed cystic cavity formation with nodules on MRI, which showed evident enhancements. All cases exhibited mutations that were predicted to activate the MAP kinase signaling pathway, including seven with BRAF p.V600E mutation and two with NF1 mutation. Five AGGs with mutations involving the MAP kinase signaling pathway also had concurrent mutations, including three with CDKN2A homozygous deletion, one with a TERT promoter mutation, one with a H3F3A mutation, and one with a PTEN mutation.Conclusions:AGG exhibits a distinct spectrum of pathology, genetic mutations and clinical behaviors, differing from GG. Given these characteristics suggest that AGG may be a distinct tumor type, further expansion of the case series is needed. Therefore, a comprehensive integration of clinical, histological, and molecular analyses is required to correctly diagnose AGG. It will also help guide treatments and prognostication.

11β-Hydroxylase deficiency(11β-OHD) is a relatively rare type of autosomal recessive hereditary disease congenital adrenal cortical hyperplasia. This article reports the diagnosis and treatment of a case of 11β-OHD combined with subacute thyroiditis leading to severe hypokalemia treated in our hospital, along with a review of relevant literature.

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.