1.Baicalein mitigates ferroptosis of neurons after subarachnoid hemorrhage
Ting ZHU ; Tingting YUE ; Yue CUI ; Yue LU ; Wei LI ; Chunhua HANG
Chinese Journal of Tissue Engineering Research 2025;29(1):52-57
BACKGROUND:Ferroptosis is a mode of programmed cell death distinct from apoptosis,necrosis,and other novel cellular deaths,which occurs mainly due to accumulated lipid peroxidation.Ferroptosis has been shown to be involved in the pathological process following subarachnoid hemorrhage.Baicalein,serving as an adept sequestered of iron,evinces its prowess by quelling lipid peroxidative cascades.Nonetheless,the enigma lingers as to whether baicalein possesses the capacity to ameliorate neuronal ferroptosis,elicited in the wake of early brain injury after subarachnoid hemorrhage. OBJECTIVE:To investigate the effect and mechanism of baicalein on neuronal ferroptosis after subarachnoid hemorrhage. METHODS:Primary neuronal cells were extracted from C57BL/6L fetal mice at 16-17 days of gestation.Hemoglobin was used to stimulate primary neuronal cells to simulate an in vitro subarachnoid hemorrhage model.The viability of primary neuronal cells treated with baicalein at concentrations of 5,15,25,50,and 100 μmol/L for 24 hours was detected by CCK-8 assay to determine the optimal concentration of baicalein.Primary neuronal cells were divided into control group,hemoglobin group,and hemoglobin+baicalein group.The levels of reactive oxygen species and malondialdehyde in cells were detected by kits.The mRNA expressions of ferroptosis-related markers PTGS2,SLC7A11,and glutathione peroxidase 4 were detected by RT-PCR.The primary neuronal cells were further divided into control group,SLC7A11 inhibitor Erastin group,hemoglobin group,hemoglobin+baicalein group,and hemoglobin+baicalein+Erastin group.The expression of the ferroptosis related markers SLC7A11 and glutathione peroxidase 4 was detected by western blot assay. RESULTS AND CONCLUSION:(1)Baicalein(25 μmol/L)was selected as the following experimental concentration.(2)Compared with the hemoglobin group,the level of malondialdehyde and the level of reactive oxygen species were significantly decreased(P<0.05)in the hemoglobin+baicalein group.(3)Compared with the hemoglobin group,the mRNA expression of PTGS2 significantly decreased,and the mRNA expression of SLC7A11 and glutathione peroxidase 4 significantly increased(P<0.000 1)in the hemoglobin+baicalein group.(4)SLC7A11 inhibitor Erastin could reverse the baicalin-improved ferroptosis effect to a certain extent(P<0.05).(5)The results showed that baicalein could alleviate the ferroptosis of neuronal cells after subarachnoid hemorrhage through the SLC7A11/GPX4 pathway.
2.Culture,identification,and differentiation of rabbit pituitary stem cells
Tingting YUE ; Ting ZHU ; Jiannan MAO ; Wei LI ; Chunhua HANG
Chinese Journal of Tissue Engineering Research 2024;28(25):3942-3946
BACKGROUND:The pituitary gland is an important endocrine organ in the body.Certain diseases can cause damage to the pituitary gland,such as pituitary adenoma and abnormal hormone secretion.Pituitary stem cells,due to their self-renewal and multi-directional differentiation potential,are expected to become a new therapeutic approach for repairing damaged pituitary glands. OBJECTIVE:To isolate and culture pituitary stem cells using the suspension cell ball culture method and identify their proliferation and differentiation ability. METHODS:Pituitary stem cells were isolated and cultured from the pituitary gland of newborn New Zealand white rabbits using the suspension cell ball culture method,and their morphological characteristics were observed.Immunofluorescence cytochemistry was used to detect the expression of pituitary stem cell markers SOX2 and Nestin.EdU labeling method was utilized to detect the proliferative ability of pituitary stem cells.After adherent and induced differentiation,the hormone levels in the culture medium were detected by ELISA. RESULTS AND CONCLUSION:Pituitary stem cell spheres could be successfully isolated by the suspension cell ball culture method,with strong proliferative ability.Positive expression of stem cell-specific markers SOX2 and Nestin was found in the cultured cells.After induction and differentiation,adrenocorticotropic hormone,thyroid hormone,growth hormone,luteinizing hormone,follicle-stimulating hormone,and prolactin levels significantly increased in the medium(P<0.001),with strong differentiation ability.
3.Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults (version 2023)
Fan FAN ; Junfeng FENG ; Xin CHEN ; Kaiwei HAN ; Xianjian HUANG ; Chuntao LI ; Ziyuan LIU ; Chunlong ZHONG ; Ligang CHEN ; Wenjin CHEN ; Bin DONG ; Jixin DUAN ; Wenhua FANG ; Guang FENG ; Guoyi GAO ; Liang GAO ; Chunhua HANG ; Lijin HE ; Lijun HOU ; Qibing HUANG ; Jiyao JIANG ; Rongcai JIANG ; Shengyong LAN ; Lihong LI ; Jinfang LIU ; Zhixiong LIU ; Zhengxiang LUO ; Rongjun QIAN ; Binghui QIU ; Hongtao QU ; Guangzhi SHI ; Kai SHU ; Haiying SUN ; Xiaoou SUN ; Ning WANG ; Qinghua WANG ; Yuhai WANG ; Junji WEI ; Xiangpin WEI ; Lixin XU ; Chaohua YANG ; Hua YANG ; Likun YANG ; Xiaofeng YANG ; Renhe YU ; Yongming ZHANG ; Weiping ZHAO
Chinese Journal of Trauma 2023;39(9):769-779
Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.
4.Dehydroepiandrosterone attenuates microglial activation and exerts neuroprotective effect after subarachnoid hemorrhage
Tao TAO ; Guangjie LIU ; Yan ZHOU ; Han WANG ; Wei LI ; Qingrong ZHANG ; Chunhua HANG
International Journal of Cerebrovascular Diseases 2020;28(2):105-112
Objective:To investigate the regulatory effect of dehydroepiandrosterone (DHEA) on the microglial activation after subarachnoid hemorrhage (SAH) in vivo and in vitro.Methods:C57BL/6 mice were used for in vivo experiments. A SAH model was induced by intravascular puncture. They were randomly divided into solvent group, model group, and DHEA pretreatment group. TUNEL staining was used to detect neuronal apoptosis level at 24 h after modeling. Iba-1/CD86 fluorescence double staining was used to detect the activation of microglia. Quantitative fluorescent polymerase chain reaction and Western blot analysis were used to detect the expression of inflammatory factors, including interleukin (IL) -1β, IL-6, tumor necrosis factor (TNF) -α, and inducible nitric oxide synthase (iNOS). The primary cultured microglia was used for in vitro experiments and it was simulated SAH by hemoglobin stimulation. They were randomly divided into control group, model group, and DHEA pretreatment group. Iba-1/CD86 fluorescence double staining was used to detect the microglial activation, and fluorescence quantitative polymerase chain reaction and Western blot analysis were used to detect the expression of inflammatory factors.Results:In vivo model experiments showed that DHEA significantly reduced neuronal apoptosis ( P<0.01) and microglial activation ( P<0.01) after SAH modeling, and IL-6 expression level significantly decreased ( P<0.01), while IL-1β, TNF-α and iNOS showed a downward trend, but there were no statistical differences. In vitro model experiments showed that DHEA could significantly inhibit microglial activation ( P<0.001) and the expression levels of various inflammatory factors ( P<0.001). Conclusions:DHEA pretreatment can reduce neuronal apoptosis and microglia activation after SAH, and it has neuroprotective effect.
5.Risk factor analysis of delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage
Jingpeng LIU ; Zhennan YE ; Xiangsheng ZHANG ; Lingyun WU ; Zihuan ZHANG ; Qiang CHEN ; Wei WU ; Lei MAO ; Xin ZHANG ; Chunhua HANG
Chinese Journal of Cerebrovascular Diseases 2017;14(1):10-14
Objective To investigate the clinical risk factors of delayed cerebral ischemia (DCI) in patients with aneurysmal subarachnoid hemorrhage (aSAH).Methods From January 2015 to April 2016,106 consecutive patients with aSAH treated with endovascular embolization at the Department of Neurosurgery,Nanjing General Hospital of Nanjing Military Command were enrolled retrospectively.The patients were divided into either a DCI group (n =34) or a non-DCI group (n =72) according to whether the occurrence of DCI.The general information of patients were collected,including sex,age,Hunt-Hess grade,modified Fisher grade,world federation of neurosurgical societies (WFNS) grade,acute cerebral edema,early (bleeding 1-3 d) hypoalbuminemia,and hypohemoglobinemia.Univariate and multivariate Logistic regression analyses were used to analyze the risk factors for occurrence of DCI.Results The incidence of DCI was 32.1% (34/106).The Hunt-Hess grade ≥ Ⅲ,modified Fisher grade ≥ Ⅲ,WFNS grade ≥ Ⅳ,acute cerebral edema,the incidences of hypoalbuminemia and hypohemoglobinemia in the DCI group were higher than those in the non-DCI group.There were significant differences between the 2 groups (all P < 0.05).There were no significant differences in the ratios of sex,age ≥ 55 years,hypertension,diabetes and hyponatremia between the 2 groups (all P > 0.05).In univariate analysis,the Hunt-Hess grade ≥ Ⅲ grade,modified Fisher grade ≥ Ⅲ,WFNS grade ≥ Ⅳ on admission,and hypoalbuminemia were used as the independent variables for multivariate analysis,the results showed that WFNS grade ≥ Ⅳ (OR,8.02;95 % CI 2.41-26.70),modified Fisher grades ≥ Ⅲ (OR,4.44;95% CI 1.38-14.32),and hypoalbuminemia at day 1-3 (OR,5.42;95% CI 1.40-20.76) were the independent risk factors for occurring DCI in patients with aSAH (all P < 0.05),and the Hunt-Hess grade ≥ Ⅲ was not the risk factor for occurring DCI in patients with aSAH (OR,1.86;95% CI 0.39-8.88,P > 0.05).Conclusion The patients with hypoalbuminemia,WFNS grade≥Ⅳ on adrnission,and modified Fisher grade ≥ Ⅲ may were the independent risk factors DCI after aSAH,and the clinical diagnosis and treatment should attach great importance to.
6.Clinical analysis and treatment of nasal complications after transsphenoidal pituitary adenomasectomy
You CHENG ; Fei XUE ; Tianyou WANG ; Junfeng JI ; Wei CHEN ; Zhiyi WANG ; Li XU ; Chunhua HANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2017;24(9):475-478
OBJECTIVE To exlpore the prevention and therapy methods of nasal complications after transsphenoidal pituitary adenomasectomy.METHODS 129 cases of pituitary adenomas underwent transsphenoidal surgery.The postoperative nasal complications were observed and disposed.RESULTS A total of 26 cases(20.1%) of nasal complications appeared in all 129 cases of pituitary adenomas underwent transsphenoidal surgery,including nasal hemorrhage 6 cases(4.8%),eerebrospinal fluid rhinorrhea 9 cases(6.9%),sphenoid sinusitis 3 cases(2.3%),atrophy rhinitis 2 cases(1.6%),olfactory dysfunction 2 cases(1.6%),nasal septum perforation 1 case(0.8%),and nasal adhesion 3 cases(2.3%).They were all cured after nasal endoscopic regular cavity clean,nasal hormone,nasal irrigation.CONCLUSION The nasal complications after transsphenoidal pituitary adenomas ectomy surgery should be timely disposed,which can effectively improve the patients' clinical symptoms.
7.Expression and prognostic value of ubiquitin carboxyl hydrolase L1 in the cerebrospinal fluid of patients with subarachnoid hemorrhage
Journal of Medical Postgraduates 2016;29(5):504-507
[Abstract ] Objective Ubiquitin carboxyl hydrolase L1(UCH-L1) is a neuron cytoplasm protein and increased UCH-L1 lev-el in cerebrospinal fluid( CSF) indicates brain injury.The study aimed to indentify UCH-L1 level in CSF of patients with subarachnoid hemorrhage(SAH) and assess the prognosis value of UCH-L1. Methods Retrospective analysis was made on 40 aneurismal SAH patients hospitalized in our department from August 2014 to February 2015.In the experiment group, CSF was taken by lumbar punc-ture in day1-day3, day5-day7, and day8-day10 after subarachnoid hemorrhage.In the control group, a single CSF sample was collect-ed during spinal anesthesia before surgery in 10 patients without neurologic disease.ELISA was applied to determine the concentration of UCH-L1 in CSF followed by comparison.Analysis was also made on relationship of the changes of UCH-L1 and modified rankins scale( MRS) 6 months later. Results CSF UCH-L1 concentrations in all SAH patients were significantly increased in day1-day3 ( P<0.05) after SAH, peaked in day5-day7 (P<0.01) and elevated un-til days 8-10 ( P<0.01 ) .MRS at 6 months showed a significant correlation with CSF levels of UCH-L1 in day5-day7 ( correlation co-efficient CC=0.691, P=0.001) and day8-day10 (CC=0.583,P=0.03), but not in days1-day3 (CC=0.364,P=0.08).SAH pa-tients with a good outcome at 6 months ( MRS<4) had much lower UCH-L1 levels in day5-day7 ( P=0.001) and day8-day10 ( P=0.041) than those with a poor outcome (MRS≥4).Higher UCH-L1 level at day5-day7 was a predictive marker of poor outcome in bi-nary logistic regression analysis (P=0.003); Conclusion The study suggests that increased CSF UCH-L1 level in day5-day7 after SAH is correlated with higher MRS and predicts an adverse clinical outcome 6 months after SAH.
8.Endovascular embolization and prognosis of middle cerebral artery aneurysms
Lili WEN ; Xin ZHANG ; Qingrong ZHANG ; Qi WU ; Xiangsheng ZHANG ; Chunhua HANG ; Jixin SHI ; Handong WANG
Chinese Journal of Cerebrovascular Diseases 2016;(2):57-61
Objective To investigate the effectiveness and safety of endovascular embolization for the treatment of middle cerebral artery aneurysms. Methods From March 2007 to May 2015,the clinical data of 170 patients with 173 middle cerebral artery aneurysms treated with endovascular embolization at the Department of Neurosurgery,Nanjing Jinling Hospital were analyzed retrospectively,including 120 (69.4%)ruptured aneurysms and 53 (30. 6%)unruptured aneurysms. The incidence of complications and prognosis were compared between the 2 groups. Results (1)A total of 170 patients who could perform endovascular embolization after preoperative evaluation successfully completed the interventional procedure. None of the patients died. (2)18 patients (10. 6%)had intraoperative and postoperative complications,including 4 (7. 5%,4/53)in the unruptured aneurysm group and 14 (11. 7%,14/120)in the ruptured aneurysm group. There was no significant difference in the incidence of complications between the two groups. (3)101 patients were followed up for 3-48 months. No new nervous system symptoms and reruptured aneurysm were observed. One patient had recurrence in the unruptured aneurysm group and 4 had recurrence in the ruptured aneurysm group. Conclusion Endovascular treatment of middle cerebral artery aneurysms is a safe,effective,and feasible technique.
9.Modified nasal septum root mucoperiosteum incision of pituitary adenomas via endonasal transsphenoidal approach
You CHENG ; Tianyou WANG ; Fei XUE ; Junfeng JI ; Li XU ; Mei ZHOU ; Chunhua HANG
Chinese Journal of Neuromedicine 2016;15(2):199-202
Objective To explore the modified nasal septum root mucoperiosteum incision of pituitary adenomas via endonasal transsphenoidal approach,and reduce the incision-related postoperative complications as rhinorrhagia,septumperforation and dysosmia.Methods Sixteen patients with pituitary adenomas,admitted to and underwent modified nasal septum root mucoperiosteum incision of pituitary adenomas via endonasal transsphenoidal approach in our hospital from January 2013 and June 2013,were chosen in our study.The postoperative nasal complications were observed.The clinical data of them were analyzed retrospectively.Results Of all the 16 patients,total or subtotal removal rate was 93.8%.Rhinorrhagia or septumperforation did not appear.Dysosmia was not appeared in 15 patients with normal olfactory function;one patient had preoperative olfactory dysfunction,without getting worsen three months after surgery.Conclusion The reformative nasoseptal root mucoperiosteum incision of transsphenoidal surgery for pituitary adenomas can maintain the integrality of sphenopalatine artery and nasoseptal olfactory mucosa,and decrease the incidence of postoperative nasal complications,such as rhinorrhagia,septumperforation and dysosmia.
10.Applicative value of three-dimensional DSA and MRI or CT fusion technology in the treatment of intracranial arteriovenous malformations
Xiangsheng ZHANG ; Xin ZHANG ; Qingrong ZHANG ; Qi WU ; Lili WEN ; Lei MAO ; Wei WU ; Chunhua HANG
Chinese Journal of Cerebrovascular Diseases 2015;(9):449-455
Objective Toinvestigatetheapplicativevalueof3D-DSAandheadMRIorCTfusion technology for guiding the individualized treatment of intracranial arteriovenous malformation (AVM ). Methods Twenty-onepatientswithAVMdiagnosedwithDSAattheDepartmentofNeurosurgery,Nanjing General Hospital of Nanjing Military Command from January 2015 to May 2015 were analyzed retrospectively. All patients performed DSA,MRI,and CT scan respectively before procedure,and they also performed 3 D-DSA and MRI or CT fusion. Of the 21 patients,15 performed MRI and 3D-DSA fusion,6 performed CT and 3D-DSA fusion. According to the image fusion results of the patients,the individualized treatment regimens were further developed,including microsurgical resection,endovascular embolization,and stereotactic radiotherapy (alone or combined treatment). The patients were followed up and observed for 2 to 6 months after procedure.Results Fromthe3D-DSAwithheadMRIorCTfusionimagesofthepatientsbeforetheprocedure not only could observe the vascular architecture of AVM,the relationship between the niduses and the surrounding nerve structures,but also could precisely locate the positions of AVM with small aneurysms or tiny AVMs. According to the results of image fusion,17 patients with AVM were treated with microsurgical resection,2 were treated with interventional embolization and stereotactic radiotherapy,and 2 were treated with stereotactic radiotherapy only. Of the 17 patients with AVM underwent microsurgical resection, none experienced intracranial rebleeding during the follow-up period. The last Glasgow outcome scale (GOS)score was 5 in 13 cases,and 4 in 4 cases. One patient with AVM underwent combined stereotactic radiotherapy had intracranial rebleeding during the follow-up period,and their last GOS score was 4. The other remaining 3 patients did not have new neurological deficits or rebleeding during the follow-up period,and theirGOSscorewas5.Conclusions 3D-DSA,headMRI,andCTfusiontechnologyarenovel, the operative method is simple,and the fusion image is accurate. They can effectively develop the individualized treatment regimens for patients with AVM.

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