Isaridin E Protects against Sepsis by Inhibiting Von Willebrand Factor-Induced Endothelial Hyperpermeability and Platelet-Endothelium Interaction.

Endothelial hyperpermeability is pivotal in sepsis-associated multi-organ dysfunction. Increased von Willebrand factor (vWF) plasma levels, stemming from activated platelets and endothelium injury during sepsis, can bind to integrin αvß3, exacerbating endothelial permeability. Hence, targeting this pathway presents a potential therapeutic avenue for sepsis. Recently, we identified isaridin E (ISE), a marine-derived fungal cyclohexadepsipeptide, as a promising antiplatelet and antithrombotic agent with a low bleeding risk. ISE's influence on septic mortality and sepsis-induced lung injury in a mouse model of sepsis, induced by caecal ligation and puncture, is investigated in this study. ISE dose-dependently improved survival rates, mitigating lung injury, thrombocytopenia, pulmonary endothelial permeability, and vascular inflammation in the mouse model. ISE markedly curtailed vWF release from activated platelets in septic mice by suppressing vesicle-associated membrane protein 8 and soluble N-ethylmaleide-sensitive factor attachment protein 23 overexpression. Moreover, ISE inhibited healthy human platelet adhesion to cultured lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs), thereby significantly decreasing vWF secretion and endothelial hyperpermeability. Using cilengitide, a selective integrin αvß3 inhibitor, it was found that ISE can improve endothelial hyperpermeability by inhibiting vWF binding to αvß3. Activation of the integrin αvß3-FAK/Src pathway likely underlies vWF-induced endothelial dysfunction in sepsis. In conclusion, ISE protects against sepsis by inhibiting endothelial hyperpermeability and platelet-endothelium interactions.

[Research progress in separation surgery combined with stereotactic radiotherapy of spinal metastases].

With the continuous improvement of cancer treatment, the survival of patients with spinal metastases has been significantly prolonged. Currently, the treatment of spinal metastases presents a trend of multi-mode. Clinical surgical methods include vertebral tumor resecting spinal canal decompression and internal fixation surgery, separation surgery, minimally invasive surgery and percutaneous ablation technology, etc. Radiotherapy techniques include traditional external radiation therapy, stereotactic radiotherapy and brachytherapy, etc. The risk of vertebral tumor resecting spinal canal decompression and internal fixation surgery, and the incidence of intraoperative and postoperative complications is high. The extension of postoperative recovery period may lead to delay of follow-up radiotherapy and other medical treatment, which has a serious impact on patients' survival and treatment confidence. However, the precision of traditional external radiation therapy is not high, and the limitation of tolerance of spinal cord makes it difficult to achieve the goal of controlling insensitive tumor. With the development of radiotherapy and surgical technology, stereotactic radiotherapy with higher accuracy and separation surgery with smaller surgical strike have become the focus of many clinical experts at present. This article reviews the progress of Hybrid treatment of separation surgery combined with stereotactic radiotherapy.

[Progress of decompression surgery for spinal metastases].

The spine is the most common site of bone metastases from malignant tumors, with metastatic epidural spinal cord compression occurring in about 10% of patients with spinal metastases. Palliative radiotherapy and simple laminectomy and decompression have been the main treatments for metastatic spinal cord compression. The former is ineffective and delayed for radiation-insensitive tumors, and the latter often impairs spinal stability. With the continuous improvement of surgical techniques and instrumentation in recent years, the treatment model of spinal metastases has changed a lot. Decompression surgery underwent open decompression, separation surgery, minimally invasive surgery and laser interintermal thermal ablation decompression. However, no matter what kind of surgical plan is adopted, it should be assessed precisely according to the specific situation of the patient to minimize the risk of surgery as far as possible to ensure the smooth follow-up radiotherapy. This paper reviews the research progress of decompression for spinal metastases.

Increased intracellular Cl- concentration mediates neutrophil extracellular traps formation in atherosclerotic cardiovascular diseases.

Neutrophil extracellular traps (NETs) play crucial roles in atherosclerotic cardiovascular diseases such as acute coronary syndrome (ACS). Our preliminary study shows that oxidized low-density lipoprotein (oxLDL)-induced NET formation is accompanied by an elevated intracellular Cl- concentration ([Cl-]i) and reduced cystic fibrosis transmembrane conductance regulator (CFTR) expression in freshly isolated human blood neutrophils. Herein we investigated whether and how [Cl-]i regulated NET formation in vitro and in vivo. We showed that neutrophil [Cl-]i and NET levels were increased in global CFTR null (Cftr-/-) mice in the resting state, which was mimicked by intravenous injection of the CFTR inhibitor, CFTRinh-172, in wild-type mice. OxLDL-induced NET formation was aggravated by defective CFTR function. Clamping [Cl-]i at high levels directly triggered NET formation. Furthermore, we demonstrated that increased [Cl-]i by CFTRinh-172 or CFTR knockout increased the phosphorylation of serum- and glucocorticoid-inducible protein kinase 1 (SGK1) and generation of intracellular reactive oxygen species in neutrophils, and promoted oxLDL-induced NET formation and pro-inflammatory cytokine production. Consistently, peripheral blood samples obtained from atherosclerotic ApoE-/- mice or stable angina (SA) and ST-elevation ACS (STE-ACS) patients exhibited increased neutrophil [Cl-]i and SGK1 activity, decreased CFTR expression, and elevated NET levels. VX-661, a CFTR corrector, reduced the NET formation in the peripheral blood sample obtained from oxLDL-injected mice, ApoE-/- atherosclerotic mice or patients with STE-ACS by lowering neutrophil [Cl-]i. These results demonstrate that elevated neutrophil [Cl-]i during the development of atherosclerosis and ACS contributes to increased NET formation via Cl--sensitive SGK1 signaling, suggesting that defective CFTR function might be a novel therapeutic target for atherosclerotic cardiovascular diseases.

Platelet CFTR inhibition enhances arterial thrombosis via increasing intracellular Cl- concentration and activation of SGK1 signaling pathway.

Platelet hyperactivity is essential for thrombus formation in coronary artery diseases (CAD). Dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) in patients with cystic fibrosis elevates intracellular Cl- levels ([Cl-]i) and enhanced platelet hyperactivity. In this study, we explored whether alteration of [Cl-]i has a pathological role in regulating platelet hyperactivity and arterial thrombosis formation. CFTR expression was significantly decreased, while [Cl-]i was increased in platelets from CAD patients. In a FeCl3-induced mouse mesenteric arteriole thrombosis model, platelet-specific Cftr-knockout and/or pre-administration of ion channel inhibitor CFTRinh-172 increased platelet [Cl-]i, which accelerated thrombus formation, enhanced platelet aggregation and ATP release, and increased P2Y12 and PAR4 expression in platelets. Conversely, Cftr-overexpressing platelets resulted in subnormal [Cl-]i, thereby decreasing thrombosis formation. Our results showed that clamping [Cl-]i at high levels or Cftr deficiency-induced [Cl-]i increasement dramatically augmented phosphorylation (Ser422) of serum and glucocorticoid-regulated kinase (SGK1), subsequently upregulated P2Y12 and PAR4 expression via NF-κB signaling. Constitutively active mutant S422D SGK1 markedly increased P2Y12 and PAR4 expression. The specific SGK1 inhibitor GSK-650394 decreased platelet aggregation in wildtype and platelet-specific Cftr knockout mice, and platelet SGK1 phosphorylation was observed in line with increased [Cl-]i and decreased CFTR expression in CAD patients. Co-transfection of S422D SGK1 and adenovirus-induced CFTR overexpression in MEG-01 cells restored platelet activation signaling cascade. Our results suggest that [Cl-]i is a novel positive regulator of platelet activation and arterial thrombus formation via the activation of a [Cl-]i-sensitive SGK1 signaling pathway. Therefore, [Cl-]i in platelets is a novel potential biomarker for platelet hyperactivity, and CFTR may be a potential therapeutic target for platelet activation in CAD.

Antiplatelet and Antithrombotic Effects of Isaridin E Isolated from the Marine-Derived Fungus via Downregulating the PI3K/Akt Signaling Pathway.

Isaridin E, a cyclodepsipeptide isolated from the marine-derived fungus Amphichorda felina (syn. Beauveria felina) SYSU-MS7908, has been demonstrated to possess anti-inflammatory and insecticidal activities. Here, we first found that isaridin E concentration-dependently inhibited ADP-induced platelet aggregation, activation, and secretion in vitro, but did not affect collagen- or thrombin-induced platelet aggregation. Furthermore, isaridin E dose-dependently reduced thrombosis formation in an FeCl3-induced mouse carotid model without increasing the bleeding time. Mechanistically, isaridin E significantly decreased the ADP-mediated phosphorylation of PI3K and Akt. In conclusion, these results suggest that isaridin E exerts potent antithrombotic effects in vivo without increasing the risk of bleeding, which may be due to its important role in inhibiting ADP-induced platelet activation, secretion and aggregation via the PI3K/Akt pathways.

[Therapy progress of spinal cord compression by metastatic spinal tumor].

Metastatic epidural compression of the spinal cord is a significant source of morbidity in patients with systemic cancer. With improvment of oncotheray, survival period in the patients is improving and metastatic cord compression is en- countered increasingly often. Surgical management performed for early circumferential decompression for the spinal cord com- pression with spine instability, and spine reconstruction performed. Patients with radiosensitive tumours without spine instabili- ty, radiotherapy is an effective therapy. Spinal stereotactic radiosurgery and minimally invasive techniques, such as vertebro- plasty and kyphoplasty, percutaneous pedicle screw fixation, radiofrequency ablation are promising options for treatment of cer- tain selected patients with spinal metastases.

[Efficacies of bone-marrow-derived mononuclear cells with a hydroxylapatite composite in the treatment of osteonecrosis of the femoral head].

OBJECTIVE: To evaluate the efficacies of core decompression and implantation of concentrated autologous bone marrow containing mononuclear cells (BMMCs) with porous hydroxylapatite composite in the treatment of osteonecrosis of the femoral head. METHODS: A total of 35 patients with 57 osteonecrosis hips with ARCO stage I, stage II and stage IIIA disease were treated by BMMCs with a porous hydroxylapatite composite. The mean age at surgery was 39.4 (26-58) years and the mean period of follow-up 28 (12-40) months. In the control group, cell-free porous hydroxylapatite composite was implanted into 17 patients (27 hips) with osteonecrosis of the femoral head and the outcomes were compared. RESULTS: At the last follow-up, postoperative Harris hip scores significantly increased in both groups (P < 0.0001). The magnitude of increase was significantly greater in the BMMCs group compared with the control group (28.3% ± 0.9% vs 18.4% ± 1.7%, P < 0.01). Postoperative visual analog scale (VAS) scores significantly decreased in both groups (P < 0.01). The magnitude of decrease was significantly greater in the BMMCs group compared with the control group (-70.2% ± 2.1% vs -51.7% ± 2.9%, P < 0.001). The clinical success rate was significantly higher in the BMMCs group compared with the control group (75.4% vs 37.0%, P < 0.01). The radiological success rates were similar between the BMMCs and control groups (59.6% vs 40.7%, P = 0.1046). CONCLUSION: The combined regimen of core decompression and implantation of concentrated autologous BMMCs with porous hydroxylapatite composite appears to confer benefits in the treatment of in stages I-IIIA osteonecrosis of the femoral head.

[Single factor analysis of motor dysfunction and imaging and clinical features in metastatic epidural spinal cord compression].

OBJECTIVE: To explore the relationship of motor dysfunction of the lower extremities with the imaging appearances and clinical features of metastatic epidural spinal cord compression (MESCCs). METHODS: From July 2006 to December 2007, 26 successive patients with metastases of the thoracic, lumbar and the cervical spine were treated in our department. Forty-three main involved vertebra in all 26 patients were evaluated by magnetic resonance imaging and computed tomography, and were scored according motor dysfunction in this study. Fourteen patients (25 vertebrae) had motor dysfunction. RESULTS: Among 26 patients, 12 cases with visceral metastasis,in which had motor dysfunction in 10 cases; 14 cases without visceral metastasis, in which had motor dysfunction in 4 cases; comparison between two groups, P=0.0079. Among vertebral presence of continuity of 43 main involved vertebrae, 16 vertebrae had motor dysfunction;among vertebral absence of continuity, motor dysfunction occurred in 9 vertebrae, comparison between two groups, P=0.1034. Among vertebral presence of lamina involvement of 43 main involved vertebrae, 11 vertebrae had motor dysfunction; among vertebral absence of lamina involvement, motor dysfunction occurred in 14 vertebrae, comparison between two groups, P=0.020 5. Among vertebral presence of protruding of vertebral posterior wall of 43 main involved vertebrae, 12 vertebrae had motor dysfunction; among vertebral absence of protruding of vertebral posterior wall, 13 vertebrae had motor dysfunction, comparison between two groups, P=0.0334. Among vertebral presence of involvement epidural space of 43 main involved vertebrae, 11 vertebrae had motor dysfunction; among vertebral absence of involvement epidural space, 14 vertebrae had motor dysfunction, comparison between two groups, P=0.003 6. Such factors as age, gender, whether or not received regular chem before admission, back pain degree of metastasis, received regular chem before admission, therapeutic efficacy of primary tumor, number of bony metastases outside spine, number of the main involved vertebrae, level of vertebral metastases location, level of continuous involved vertebrae, vertebral-body involvement, fracture of anterior column, fracture of posterior wall, and pedicle involvement had no effects on incidence of motor dysfunction due to MESCC (P>0.05). CONCLUSION: MESCC with visceral metastases, lamina involvement, presence of outstanding buttocks sign of posterior wall,involvement epidural space tended to cause symptomatic MESCC. Incidence of continuity of main involved vertebrae occurred more frequently in the CUTS compared with other levels of spine.

[Relationship of motor deficits and imaging features in metastatic epidural spinal cord compression].

OBJECTIVE: To explore the relationship of motor deficits of the lower extremities with the imaging features of malignant spinal cord compression (MESCCs). METHODS: From July 2006 through December 2008, 56 successive MESCC patients were treated at our department. All were evaluated by magnetic resonance imaging and computed tomography and were scored according to motor deficits Frankel grading on admission. Imaging assessment factors of main involved vertebrae were level of vertebral metastatic location, epidural space involvement, vertebral body involvement, lamina involvement, posterior protrusion of posterior wall, pedicle involvement, continuity of main involved vertebrae, fracture of anterior column, fracture of posterior wall, location in upper thoracic spine and/or cervicothoracic junction. RESULTS: Occurrence was the same between paralytic state of MESCCs and epidural space involvement of imaging features. Multiple regression equation showed that paralytic state had a linear regression relationship with imaging factors of lamina involvement (X1), posterior protrusion of posterior wall (X2), location in upper thoracic spine and/or cervicothoracic junction (X7) of main involved vertebrae. The optimal regression equation of paralytic state (Y) and imaging feature (X) was Y = -0.009 +0.639X, + 0.149X, +0.282X. Lamina involvement of main involved vertebrae has a greatest influence upon paralytic state of MESCC patients. CONCLUSIONS: Imaging factors of lamina involvement, posterior protrusion of posterior wall, location in upper thoracic spine and/or cervicothoracic junction of main involved vertebrae can predict the paralytic state of MESCC patients. MESCC with lamina involvement is more easily encroached on epidural space.

[Intraarterial neoadjuvant chemotherapy for extremity osteosarcoma].

OBJECTIVE: To summarize the experiences in intraarterial neoadjuvant chemotherapy for extremity osteosarcoma. METHODS: Between January 2002 and December 2007, 111 patients with stage IIB extremity osteosarcoma received preoperative intraarterial chemotherapy after placing chemotherapy pump subcutaneously, en bloc resection and postoperative adjuvant chemotherapy. There were 63 males and 48 females with an average age of 18 years old (range: 14 - 39). The time from symptom onset to hospitalization varied from several days to 6 months. The induction chemotherapy regimen included epirubicin [50 - 70 mg/m(2) by 4-hour intraarterial infusion/day for 3 days] and epirubicin plus adriamycin [100 - 120 mg/m(2) by 2-hour intraarterial infusion/day for 3 days] repetitively every 2 - 3 weeks. Among which 24 cases only received 2 cycles of induction chemotherapy was assigned into the nonstandard chemotherapy group and 87 cases receiving 3 - 6 cycles of induction chemotherapy the standard chemotherapy group. The number of preoperative chemotherapeutical cycles of standard chemotherapy group depended on the clinical and radiographic evaluation of chemotherapy efficacy. RESULTS: The median follow-up time was 28 (8 - 48) months. The rate of extremity preservation surgery was 89.53% (77/86) in the standard chemotherapy group and 37.5% (9/24) in the nonstandard chemotherapy group. Kaplan-Meier survival analysis showed that the 3-year overall survival rate and disease-free survival rate of all 111 cases were 68.3% and 65.9% respectively. There were significant differences in overall survival rate (38.9%, 80.0%, P = 0.000), disease-free survival rate (30.1%, 79.5%, P = 0.000), distant metastatic rate (66.67%, 16.09%, P = 0.0000) and local recurrence rate (58.33%, 13.79%, P = 0.0000) between two groups. CONCLUSION: Standard intraarterial neo-adjuvant chemotherapy is more effective than nonstandard intraarterial induction chemotherapy to treat stage IIB extremity osteosarcoma.

[Analysis of lumbar disc degeneration using three-dimensional nonlinear finite element method].

OBJECTIVE: To explore the effects of intervertebral disc degeneration on the biomechanical behavior of the lumbar motion segment. METHODS: A three-dimensional nonlinear finite element model of L4-L5 segment was established using CAD technique based on CT images. A normal disc model and three degenerative disc models were established by changing the parameters such as disc material properties and disc height. The effects of disc degeneration on the biomechanical properties, including stiffness, nucleus pressure, maximum von Mises stress in the annulus, and force of posterior structure were studied under two moment loads (flexion and extension) and for three different direct forces (compression, and anterior and posterior shear forces), and the stress distribution of vertebral body and end- plate was also analyzed. RESULTS: The stiffness of the lightly degenerative disc model was decreased compared with that of the normal disc, while it was increased in the moderately and severely degenerative disc models when compared with the normal disc. The force of posterior structure in the lightly degenerative disc model was increased while the values of force of posterior structure in the moderately and severely degenerative disc models were decreased gradually. The maximum von Mises stress in the annulus increased, and the nucleus pressure decreased as the disc degeneration progressed. Moreover, the stress of intervertebral disc, vertebral body, and end-plate was distributed more peripherally. CONCLUSION: Light degeneration of intervertebral disc leads to instability of lumbar spine, while the stability restores with further degeneration of disc. There is a negative correlation between the force of posterior structure and the load on the intervertebral disc. With the disc degeneration progressing, the intervertebral disc load pattern changes, the stress of intervertebral disc, vertebral body, and end-plate is concentrated peripherally.

[Diagnosis and treatment of odontoid fracture combined with lower cervical spinal injury].

OBJECTIVE: To investigate the mechanism, clinical features and treatment of odontoid fracture combined with lower cervical spinal injury. METHODS: From January 1999 to December 2004, 57 cases of type II or shallow type III odontoid fractures were studied retrospectively. Six cases were found combined with lower cervical injury, the mean age was 54 years, and 4 of the 6 cases were complicated with cervical spondylarthrosis or ankylosing spondylitis. For the lower cervical injury, fracture-dislocation was found in 2 cases, the disruption of disc and ligament was found in 4 cases among which 2 cases were suffered from incomplete spinal cord injury; Both were caused by lower cervical spinal injury. All of the 6 cases were performed with surgery in odontoid fracture and lower cervical spinal injury simultaneously; Lower cervical spinal injuries were stabilized firstly in 2 cases, which responsible for neurological involvement; For the other 4 cases without neurological involvement, stabilization was performed in odontoid fracture firstly in 2 cases, due to inability to achieve reduction of odontoid fracture preoperatively, however, for the another 2 cases with anatomic reduction of the odontoid fracture preoperatively, lower cervical injuries were stabilized firstly. RESULTS: After an average follow-up of 10 months, all cases were obtained solid fusion both in odontoid fracture and lower cervical spinal injury, and without the complications associated with operation and prolonged bed rest. Two cases with neurological defect improved 1 scale in Frankel score. CONCLUSIONS: The incidence of odontoid fracture combined with lower cervical spinal injury is about 10.5% of the odontoid fracture, and it is vulnerable in the elderly patient with cervical spondylarthrosis. MRI should be used routinely for accurate diagnosis. Surgical stabilization is the choice of treatment due to facilitating early rehabilitation and reducing the complications. The surgical schedule is planned according to the fact of neurological involvement and the extent of stability between the odontoid fracture and lower cervical spinal injury.