Background: Diabetic pain patients have increased pain at night. Exosomes can relieve neuropathic pain. This study aimed to investigate the efficacy of exosome administration at different time points in relieving diabetic neuropathic pain (DNP) in rats. Methods: M2 macrophages from bone marrow were induced in mice and exosomes were extracted. A diabetic rat model was induced using streptozotocin, with the mechanical withdrawal threshold (MWT) of the rats beingmeasured at ≤ 80% of the basal value after 14 days, indicating successful construction of the DNP rat model.Exosomes were administered on three consecutive days at ZT0 (zeitgeber time) and ZT12. Parameters including blood glucose levels, body weight, MWT, and thermal withdrawal latency (TWL) were assessed in the rats. The lumbar spinal cord of rats was examined on days 21 and 28 to measure inflammatory factors and observe the expression of M1 and M2 microglia. Furthermore, microglia were exposed to lipopolysaccharide (LPS) and LPS + exosomes in a controlled in vitro setting to assess alterations in microglia phenotype involving the NF-kB p65 andIKBα inflammatory signaling pathways. Results: The findings revealed that administration of exosomes during the rat resting period at ZT12 resulted in increased MWT and TWL, as well as a shift in microglia polarization towards the M2 phenotype. In vitro analysis indicated that exosomes influenced microglia polarization and suppressed the phosphorylation of NF-kB p65 andIKBα.  Conclusions Temporal therapy with exosomes effectively reduces pain in DNP rats by polarizing microglia andaffecting NF-kB p65 and IKBα signaling pathways.

Background: Diabetic pain patients have increased pain at night. Exosomes can relieve neuropathic pain. This study aimed to investigate the efficacy of exosome administration at different time points in relieving diabetic neuropathic pain (DNP) in rats. Methods: M2 macrophages from bone marrow were induced in mice and exosomes were extracted. A diabetic rat model was induced using streptozotocin, with the mechanical withdrawal threshold (MWT) of the rats beingmeasured at ≤ 80% of the basal value after 14 days, indicating successful construction of the DNP rat model.Exosomes were administered on three consecutive days at ZT0 (zeitgeber time) and ZT12. Parameters including blood glucose levels, body weight, MWT, and thermal withdrawal latency (TWL) were assessed in the rats. The lumbar spinal cord of rats was examined on days 21 and 28 to measure inflammatory factors and observe the expression of M1 and M2 microglia. Furthermore, microglia were exposed to lipopolysaccharide (LPS) and LPS + exosomes in a controlled in vitro setting to assess alterations in microglia phenotype involving the NF-kB p65 andIKBα inflammatory signaling pathways. Results: The findings revealed that administration of exosomes during the rat resting period at ZT12 resulted in increased MWT and TWL, as well as a shift in microglia polarization towards the M2 phenotype. In vitro analysis indicated that exosomes influenced microglia polarization and suppressed the phosphorylation of NF-kB p65 andIKBα.  Conclusions Temporal therapy with exosomes effectively reduces pain in DNP rats by polarizing microglia andaffecting NF-kB p65 and IKBα signaling pathways.

Background: Diabetic pain patients have increased pain at night. Exosomes can relieve neuropathic pain. This study aimed to investigate the efficacy of exosome administration at different time points in relieving diabetic neuropathic pain (DNP) in rats. Methods: M2 macrophages from bone marrow were induced in mice and exosomes were extracted. A diabetic rat model was induced using streptozotocin, with the mechanical withdrawal threshold (MWT) of the rats beingmeasured at ≤ 80% of the basal value after 14 days, indicating successful construction of the DNP rat model.Exosomes were administered on three consecutive days at ZT0 (zeitgeber time) and ZT12. Parameters including blood glucose levels, body weight, MWT, and thermal withdrawal latency (TWL) were assessed in the rats. The lumbar spinal cord of rats was examined on days 21 and 28 to measure inflammatory factors and observe the expression of M1 and M2 microglia. Furthermore, microglia were exposed to lipopolysaccharide (LPS) and LPS + exosomes in a controlled in vitro setting to assess alterations in microglia phenotype involving the NF-kB p65 andIKBα inflammatory signaling pathways. Results: The findings revealed that administration of exosomes during the rat resting period at ZT12 resulted in increased MWT and TWL, as well as a shift in microglia polarization towards the M2 phenotype. In vitro analysis indicated that exosomes influenced microglia polarization and suppressed the phosphorylation of NF-kB p65 andIKBα.  Conclusions Temporal therapy with exosomes effectively reduces pain in DNP rats by polarizing microglia andaffecting NF-kB p65 and IKBα signaling pathways.

Background: Diabetic pain patients have increased pain at night. Exosomes can relieve neuropathic pain. This study aimed to investigate the efficacy of exosome administration at different time points in relieving diabetic neuropathic pain (DNP) in rats. Methods: M2 macrophages from bone marrow were induced in mice and exosomes were extracted. A diabetic rat model was induced using streptozotocin, with the mechanical withdrawal threshold (MWT) of the rats beingmeasured at ≤ 80% of the basal value after 14 days, indicating successful construction of the DNP rat model.Exosomes were administered on three consecutive days at ZT0 (zeitgeber time) and ZT12. Parameters including blood glucose levels, body weight, MWT, and thermal withdrawal latency (TWL) were assessed in the rats. The lumbar spinal cord of rats was examined on days 21 and 28 to measure inflammatory factors and observe the expression of M1 and M2 microglia. Furthermore, microglia were exposed to lipopolysaccharide (LPS) and LPS + exosomes in a controlled in vitro setting to assess alterations in microglia phenotype involving the NF-kB p65 andIKBα inflammatory signaling pathways. Results: The findings revealed that administration of exosomes during the rat resting period at ZT12 resulted in increased MWT and TWL, as well as a shift in microglia polarization towards the M2 phenotype. In vitro analysis indicated that exosomes influenced microglia polarization and suppressed the phosphorylation of NF-kB p65 andIKBα.  Conclusions Temporal therapy with exosomes effectively reduces pain in DNP rats by polarizing microglia andaffecting NF-kB p65 and IKBα signaling pathways.

Background: Diabetic pain patients have increased pain at night. Exosomes can relieve neuropathic pain. This study aimed to investigate the efficacy of exosome administration at different time points in relieving diabetic neuropathic pain (DNP) in rats. Methods: M2 macrophages from bone marrow were induced in mice and exosomes were extracted. A diabetic rat model was induced using streptozotocin, with the mechanical withdrawal threshold (MWT) of the rats beingmeasured at ≤ 80% of the basal value after 14 days, indicating successful construction of the DNP rat model.Exosomes were administered on three consecutive days at ZT0 (zeitgeber time) and ZT12. Parameters including blood glucose levels, body weight, MWT, and thermal withdrawal latency (TWL) were assessed in the rats. The lumbar spinal cord of rats was examined on days 21 and 28 to measure inflammatory factors and observe the expression of M1 and M2 microglia. Furthermore, microglia were exposed to lipopolysaccharide (LPS) and LPS + exosomes in a controlled in vitro setting to assess alterations in microglia phenotype involving the NF-kB p65 andIKBα inflammatory signaling pathways. Results: The findings revealed that administration of exosomes during the rat resting period at ZT12 resulted in increased MWT and TWL, as well as a shift in microglia polarization towards the M2 phenotype. In vitro analysis indicated that exosomes influenced microglia polarization and suppressed the phosphorylation of NF-kB p65 andIKBα.  Conclusions Temporal therapy with exosomes effectively reduces pain in DNP rats by polarizing microglia andaffecting NF-kB p65 and IKBα signaling pathways.

OBJECTIVE: To precise classify sacral meningeal cysts, effective guide minimally invasive neurosurgery and postoperative personalized rehabilitation by multiple dimensions radiographic reconstruction MRI. METHODS: From March to December 2021, based on the original 3D-fast imaging employing steadystate acquisition (FIESTA) scanning sequence, 92 patients with sacral meningeal cysts were pre-operatively evaluated by multiple dimensional reconstruction MRI. The shape of nerve root and the leakage of cyst were reconstructed according to the direction of nerve root or leakage track showed on original MRI scans. Sacral canal cysts were accurately classified as including nerve root and without nerve root, so as to accurately design the incision of skin and formulate corresponding open range of the posterior wall of the sacral canal. Under the microscope intraoperation, the shape of the nerve roots inside cysts or leakage track of the cysts without nerve roots were verified and explored. After the reinforcement and shaping operation, several reexaminations of multiple dimensional reconstruction MRI were performed to understand the deformation of the nerve root and hydrops in the operation cavity, so as to formulate a persona-lized rehabilitation plan for the patients. RESULTS: Among the 92 patients with sacral mengingeal cyst, 58 (63.0%) cysts with nerve root cyst, 29 (31.5%) cysts without nerve root cyst, and 5 (5.4%) cysts with mixed sacral canal cyst. In 58 patients with nerve root cysts, the accuracy of preoperative clinical classification on MRI image reached 96.6% (56/58) through confirmation by operating microscope. Only 2 cases of large single cyst with nerve root on the head of cyst were mistaken for without nerve root type. In 29 patients with sacral cyst without nerve root, the accuracy of preoperative image reached 100% through confirmation by operating microscope. The accuracy of judging the internal nerve root and leakage of 12 cases with recurrent sacral cyst was also 100%. Two cases of delayed postoperative hydrops were found one month after operation. After rehabilitation treatment by moxibustion and bathing, the hydrops disappeared 4-6 months after operation. CONCLUSION Multiple dimensional reconstruction MRI can precisely make clinical classification of sacral meningeal cysts before operation, guide minimally invasive neurosurgery effectively, and improve the rehabilitation effect.
Humans ; Magnetic Resonance Imaging/methods* ; Male ; Female ; Sacrum/surgery* ; Adult ; Middle Aged ; Imaging, Three-Dimensional/methods* ; Cysts/rehabilitation* ; Aged ; Adolescent ; Young Adult ; Spinal Nerve Roots/diagnostic imaging* ; Minimally Invasive Surgical Procedures ; Neurosurgical Procedures/methods*

BACKGROUND:Much of the current research on M2 macrophage-derived exosomes focuses on their effects on wound healing and osteoblast proliferation and differentiation,while few studies have focused on their role in regulating microglia phenotype.OBJECTIVE:To discuss the role and molecular mechanisms of M2 macrophage-derived exosomes in the phenotypic regulation of microglia.MERHODS:(1)Bone marrow primary macrophages were extracted and then stimulated with 50 ng/mL interleukin 4 for 24 hours to promote macrophage M2-type polarization.Flow cytometry and cellular immunofluorescence were used to identify the M2-type macrophage marker CD206.(2)M2 macrophage-derived exosomes were extracted and identified.(3)Microglia BV2 were randomly divided into three groups:control group,lipopolysaccharide group,and treatment group.No treatment was done in the control group.500 ng/mL lipopolysaccharide was added to the intervention for 24 hours in the lipopolysaccharide group.500 ng/mL lipopolysaccharide and 25 μg/mL M2 macrophage-derived exosomes were added simultaneously to the treatment group for 24 hours.ELISA was performed to detect the secretion of tumor necrosis factor α and interleukin 10 in the culture supernatant.qRT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase,arginase 1,interleukin 1β,and interleukin 10 in the cells.Western blot assay was performed to detect the protein expression of inducible nitric oxide synthase,arginase 1,and nuclear factor-κB signaling pathway related protein expression.RESULTS AND CONCLUSION:(1)ELISA results showed that the secretion of tumor necrosis factor α was significantly increased in the lipopolysaccharide group compared with the control group.The secretion of tumor necrosis factor α was reduced and the secretion of interleukin 10 was increased in the treatment group compared with the lipopolysaccharide group.(2)The qRT-PCR results showed that compared with the control group,the mRNA expression of interleukin 1β and inducible nitric oxide synthase increased in the lipopolysaccharide group.Compared with the lipopolysaccharide group,the mRNA expression of interleukin 1β and inducible nitric oxide synthase decreased,and the mRNA expression of interleukin 10 and arginase 1 increased in the treatment group.(3)Western blot assay results showed that the expression of inducible nitric oxide synthase protein was increased in the lipopolysaccharide group compared with the control group.The expression of inducible nitric oxide synthase protein was decreased and the expression of arginase 1 protein was elevated in the treatment group compared with the lipopolysaccharide group.(4)Compared with the control group,the expression of p65 and p-IκB-α proteins in the nuclear factor-κB signaling pathway was reduced in the lipopolysaccharide group,whereas the expression of p65 and p-IκB-α proteins was elevated in the treatment group compared with the lipopolysaccharide group.The results showed that M2-type macrophage-derived exosomes could significantly inhibit lipopolysaccharide-induced inflammatory responses in microglia,enhance the expression of the anti-inflammatory factor interleukin 10,suppress the expression of the pro-inflammatory factors tumor necrosis factor α and interleukin 1β,and promote microglial cell phenotypes polarized from the M1-type to the M2-type.The mechanism may be related to the inhibition of nuclear factor-κB signaling pathway activation by M2-type macrophage-derived exosomes.

BACKGROUND:Much of the current research on M2 macrophage-derived exosomes focuses on their effects on wound healing and osteoblast proliferation and differentiation,while few studies have focused on their role in regulating microglia phenotype.OBJECTIVE:To discuss the role and molecular mechanisms of M2 macrophage-derived exosomes in the phenotypic regulation of microglia.MERHODS:(1)Bone marrow primary macrophages were extracted and then stimulated with 50 ng/mL interleukin 4 for 24 hours to promote macrophage M2-type polarization.Flow cytometry and cellular immunofluorescence were used to identify the M2-type macrophage marker CD206.(2)M2 macrophage-derived exosomes were extracted and identified.(3)Microglia BV2 were randomly divided into three groups:control group,lipopolysaccharide group,and treatment group.No treatment was done in the control group.500 ng/mL lipopolysaccharide was added to the intervention for 24 hours in the lipopolysaccharide group.500 ng/mL lipopolysaccharide and 25 μg/mL M2 macrophage-derived exosomes were added simultaneously to the treatment group for 24 hours.ELISA was performed to detect the secretion of tumor necrosis factor α and interleukin 10 in the culture supernatant.qRT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase,arginase 1,interleukin 1β,and interleukin 10 in the cells.Western blot assay was performed to detect the protein expression of inducible nitric oxide synthase,arginase 1,and nuclear factor-κB signaling pathway related protein expression.RESULTS AND CONCLUSION:(1)ELISA results showed that the secretion of tumor necrosis factor α was significantly increased in the lipopolysaccharide group compared with the control group.The secretion of tumor necrosis factor α was reduced and the secretion of interleukin 10 was increased in the treatment group compared with the lipopolysaccharide group.(2)The qRT-PCR results showed that compared with the control group,the mRNA expression of interleukin 1β and inducible nitric oxide synthase increased in the lipopolysaccharide group.Compared with the lipopolysaccharide group,the mRNA expression of interleukin 1β and inducible nitric oxide synthase decreased,and the mRNA expression of interleukin 10 and arginase 1 increased in the treatment group.(3)Western blot assay results showed that the expression of inducible nitric oxide synthase protein was increased in the lipopolysaccharide group compared with the control group.The expression of inducible nitric oxide synthase protein was decreased and the expression of arginase 1 protein was elevated in the treatment group compared with the lipopolysaccharide group.(4)Compared with the control group,the expression of p65 and p-IκB-α proteins in the nuclear factor-κB signaling pathway was reduced in the lipopolysaccharide group,whereas the expression of p65 and p-IκB-α proteins was elevated in the treatment group compared with the lipopolysaccharide group.The results showed that M2-type macrophage-derived exosomes could significantly inhibit lipopolysaccharide-induced inflammatory responses in microglia,enhance the expression of the anti-inflammatory factor interleukin 10,suppress the expression of the pro-inflammatory factors tumor necrosis factor α and interleukin 1β,and promote microglial cell phenotypes polarized from the M1-type to the M2-type.The mechanism may be related to the inhibition of nuclear factor-κB signaling pathway activation by M2-type macrophage-derived exosomes.

Objective To investigate the serum levels of semaphorin 3E(Sema3E)in patients with intracranial aneurysms,revealing the correlation between Sema3E and 1-month poor prognosis after interventional embolization.Methods This study was a prospective single-center cohort study,recruiting 102 consecutive patients with intracranial aneurysms who underwent interventional surgery from June 2020 to January 2022 in our hospital.Among them,11 patients were excluded.Clinical and radiological profiles were collected.Peripheral blood was collected after admission,and serum Sema3E levels were determined by enzyme-linked immunosorbent assay.All the aneurysms were treated with endovascular coil embolization or stent-assisted coil embolization.The primary outcome was evaluated with the Glasgow Outcome Scale(GOS)1 month after interventional therapy.The favorable outcome was defined as a GOS score of 4-5,and a poor outcome was defined as a GOS score of 1-3(severe disability,vegetative state,or death).Univariate and multivariate logistic regression analyses were used to identify potential prognostic factors after interventional therapy.Results The average age of 91 patients with intracranial aneurysm was 59.9±11.0 years old,including 70 cases(76.9%)with favorable prognosis and 21 cases(23.1%)with poor prognosis.The mean preoperative Glasgow Coma Scale(GCS)score of the poor prognosis group(9.4±4.5)was significantly lower than that of the favorable prognosis group(13.3±2.5;P<0.001).In the poor prognosis group,the Hunt-Hess grade(3.6±0.6 vs.2.0±1.3,P<0.001)and the serum Sema3E levels[(6.21±1.58)μg/L vs.(4.38±1.77)μg/L,P<0.001]were significantly higher than those in the favorable prognosis group.Logistic regression analysis showed the Hunt-Hess grade(OR =7.150,P =0.003),stent-assisted coil embolization(OR =15.777,P =0.010),and the serum Sema3E level(OR =1.756,P =0.027)were independent prognostic factors for intracranial aneurysms after interventional therapy.Conclusions The serum Sema3E level is closely correlated with the severity of intracranial aneurysms.The serum Sema3E level is a prognostic factor for interventional treatment,which can be used as a biomarker for predicting poor outcomes.

Objective:To investigate the alterations of cholinergic and monoamine neurotransmitters in cerebrospinal fluid in patients with postoperative delirium.Methods:Sixty-eight patients with normal preoperative cognitive functions were enrolled.They were diagnosed with spinal extramedullary intradural space-occupying lesion and underwent surgical resection.Among these patients, 18 developed postoperative delirium (delirium group), 46 had no postoperative delirium (control group), and the diagnosis of delirium was unclear in four cases (excluded). Cerebrospinal fluid (CSF) was collected before the surgical resection of lesions and on the third day postoperatively.The concentrations of acetylcholine (Ach), norepinephrine (NE), adrenaline (E), dopamine (DA) and serotonin (5-HT) were measured using the high-performance liquid chromatography/electrochemical method.Results:Prior to the surgical resection, there were no significant differences in the Ach, NE, E, DA or 5-HT baseline levels in the CSF between the delirium group and the control group (all P>0.05). After surgery, the Ach level in the delirium group ((0.63±0.26) μmol/L) was significantly lower than that in the control group ((0.77±0.19) μmol/L) ( P=0.032), and there were no significant differences in other neurotransmitter levels (all P>0.05). In the delirium group, the level of Ach in the CSF after surgery ((0.63±0.26) μmol/L) was significantly lower than the baseline level ((0.75±0.19) μmol/L) ( P=0.021). The postoperative NE level ((1.58±0.28) μmol/L) was significantly higher than the baseline level ((1.49±0.21) μmol/L) ( P=0.036). There was no significant difference in the adrenaline level ( P=0.497). The postoperative DA level ((0.86±0.18) μmol/L) was significantly higher than the baseline level ((0.82±0.15) μmol/L) ( P=0.045), and the postoperative 5-HT level ((2.94±0.28) μmol/L) was also significantly higher than the baseline level ((2.75±0.35) μmol/L) ( P=0.022). In the control group, only the postoperative 5-HT level ((2.90±0.31) μmol/L) was significantly higher than the baseline level ((2.76±0.26) μmol/L) ( P=0.016), while the postoperative levels of other neurotransmitters were not significantly changed when compared to the baseline levels (all P>0.05). Conclusion:The cholinergic neurotransmitter levels were reduced while the monoamine neurotransmitter levels were increased in the cerebrospinal fluid in patients with postoperative delirium, which suggests that cholinergic hypoactivity and monoaminergic hyperexcitability may be important pathophysiological processes in the occurrence and development of postoperative delirium.