Role of Galectin-3 in intervertebral disc degeneration: an experimental study.

BACKGROUND: This study investigated the role of Galectin-3 in the degeneration of intervertebral disc cartilage. METHODS: The patients who underwent lumbar spine surgery due to degenerative disc disease were recruited and divided into Modic I, Modic II, and Modic III; groups. HE staining was used to detect the pathological changes in endplates. The changes of Galectin-3, MMP3, Aggrecan, CCL3, and Col II were detected by immunohistochemistry, RT-PCR, and Western blot. MTT and flow cytometry were used to detect cartilage endplate cell proliferation, cell cycle, and apoptosis. RESULTS: With the progression of degeneration (from Modic I to III), the chondrocytes and density of the cartilage endplate of the intervertebral disc decreased, and the collagen arrangement of the cartilage endplate of the intervertebral disc was broken and calcified. Meanwhile, the expressions of Aggrecan, Col II, Galectin-3, Aggrecan, and CCL3 gradually decreased. After treatment with Galectin-3 inhibitor GB1107, the proliferation of rat cartilage end plate cells was significantly reduced (P < 0.05). GB1107 (25 µmol/L) also significantly promoted the apoptosis of cartilage endplate cells (P < 0.05). Moreover, the percentage of cartilage endplate cells in the G1 phase was significantly higher, while that in the G2 and S phases was significantly lower (P < 0.05). Additionally, the mRNA and protein expression levels of MMP3, CCL3, and Aggrecan in rat cartilage end plate cells were lower than those in the control group. CONCLUSIONS: Galectin-3 decreases with the progression of the cartilage endplate degeneration of the intervertebral disc. Galectin-3 may affect intervertebral disc degeneration by regulating the degradation of the extracellular matrix.

K63 Ubiquitination of P21 Can Facilitate Pellino-1 in the Context of Chronic Obstructive Pulmonary Disease and Lung Cellular Senescence.

Chronic obstructive pulmonary diseases (COPD) is a kind of age-related, airflow-obstruction disease mostly caused by cigarette smoke. However, the relationship between COPD and lung cellular senescence is still not fully understood. Here, we found silencing Pellino-1 could inhibit the protein level of P21. Then, through constructing cell lines expressed ubiquitin-HA, we found that the E3 ubiquitin ligase Pellino-1 could bind to senescence marker p21 and modify p21 by K63-site ubiquitination by co-IP assays. Furthermore, we found that p21-mediated lung cellular senescence could be inhibited by silencing Pellino-1 in a D-galactose senescence mice model. Moreover, by constructing a COPD mouse model with shPellino-1 adenovirus, we found that silencing Pellino-1 could inhibit COPD and inflammation via reduction of SASPs regulated by p21. Taken together, our study findings elucidated that silencing E3 ligase Pellino-1 exhibits therapeutic potential for treatment to attenuate the progression of lung cellular senescence and COPD.

Saquayamycin B1 Suppresses Proliferation, Invasion, and Migration by Inhibiting PI3K/AKT Signaling Pathway in Human Colorectal Cancer Cells.

Moromycin B (Mor B), saquayamycin B1 (Saq B1), saquayamycin B (Saq B), and landomycin N (Lan N), four angucyclines produced by the marine-derived actinomycete Streptomyces sp., are a class of polyketone compounds containing benzanthracene. Here, the structure-activity relationship of these four compounds was analyzed in human colorectal cancer (CRC) cells. Saq B1, which showed the strongest cytotoxicity with an IC50 of 0.18-0.84 µM for CRC cells in MTT assays, was employed to test underlying mechanisms of action in SW480 and SW620 cells (two invasive CRC cell lines). Our results showed that Saq B1 inhibited CRC cell proliferation in a dose- and time-dependent manner. Notably, lower cytotoxicity was measured in normal human hepatocyte cells (QSG-7701). Furthermore, we observed proapoptosis, antimigration, and anti-invasion activities of Saq B1 in CRC cells. At the same time, the protein and mRNA expression of important markers related to the epithelial-mesenchymal transition (EMT) and apoptosis changed, including N-cadherin, E-cadherin, and Bcl-2, in Saq B1-treated CRC cells. Surprisingly, the PI3K/AKT signaling pathway was shown to be involved in Saq B1-induced apoptosis, and in inhibiting invasion and migration. Computer docking models also suggested that Saq B1 might bind to PI3Kα. Collectively, these results indicate that Saq B1 effectively inhibited growth and decreased the motor ability of CRC cells by regulating the PI3K/AKT signaling pathway, which provides more possibilities for the development of drugs in the treatment of CRC.

Natural Marine Products: Anti-Colorectal Cancer In Vitro and In Vivo.

Colorectal cancer, a malignant tumor with high mortality, has a poor prognosis due to drug resistance and toxicity in clinical surgery and chemotherapy. Thus, finding safer and more efficient drugs for clinical trials is vital and urgent. Natural marine compounds, with rich resources and original chemical structures, are applied widely in anticancer treatments. We provide a systematic overview of recently reported marine compounds such as alkaloids, peptides, terpenoids, polysaccharides, and carotenoids from in vitro, in vivo, and clinical studies. The in vitro studies summarized the marine origins and pharmacological mechanisms, including anti-proliferation, anti-angiogenesis, anti-migration, anti-invasion, the acceleration of cycle arrest, and the promotion of tumor apoptosis, of various compounds. The in vivo studies outlined the antitumor effects of marine compounds on colorectal cancer model mice and evaluated their efficacy in terms of tumor inhibition, hepatotoxicity, and nephrotoxicity. The clinical studies summarized the major chemical classifications and targets of action of the clinical drugs that have entered clinical approval and completed approval for marine anticancer. In summary, we present the current situation regarding the application of natural anti-colorectal cancer marine compounds and prospects for their clinical application.

Actinomycin V Induces Apoptosis Associated with Mitochondrial and PI3K/AKT Pathways in Human CRC Cells.

Actinomycin (Act) V, an analogue of Act D, presented stronger antitumor activity and less hepatorenal toxicity than Act D in our previous studies, which is worthy of further investigation. We hereby report that Act V induces apoptosis via mitochondrial and PI3K/AKT pathways in colorectal cancer (CRC) cells. Act V-induced apoptosis was characterized by mitochondrial dysfunction, with loss of mitochondria membrane potential (MMP) and cytochrome c release, which then activated cleaved caspase-9, cleaved caspase-3, and cleaved PARP, revealing that it was related to the mitochondrial pathway, and the apoptotic trendency can be reversed by caspase inhibitor Z-VAD-FMK. Furthermore, we proved that Act V significantly inhibited PI3K/AKT signalling in HCT-116 cells using cell experiments in vitro, and it also presented a potential targeted PI3Kα inhibition using computer docking models. Further elucidation revealed that it exhibited a 28-fold greater potency than the PI3K inhibitor LY294002 on PI3K inhibition efficacy. Taken together, Act V, as a superior potential replacement of Act D, is a potential candidate for inhibiting the PI3K/AKT pathway and is worthy of more pre-clinical studies in the therapy of CRC.

Resistomycin Induced Apoptosis and Cycle Arrest in Human Hepatocellular Carcinoma Cells by Activating p38 MAPK Pathway In Vitro and In Vivo.

Resistomycin, a quinone-related natural antibiotic, has shown strong inhibitory activity against human hepatocellular carcinoma (HCC) in vitro. Here, we investigated the role of p38 MAPK in the pro-apoptotic and G2/M phase arrest action of HCC HepG2 cells upon treatment with resistomycin in vitro and in vivo. Our results showed that resistomycin dose- and time-dependently reduced the viability of HepG2 cells and also showed lower cytotoxicity in normal human kidney cells (293T) and hepatocyte cells (HL-7702). Resistomycin treatment induced apoptosis and cell cycle arrest in HepG2 cells, accompanied by changes in the expression of related proteins, including Bax, Cyclin B1, etc. Surprisingly, resistomycin-mediated apoptotic cell death and cell cycle arrest were impeded by SB203580 (an inhibitor of p38 catalytic activity), suggesting that p38 MAPK signaling may play an important role that impedes eventual cell death. In this connection, data in vitro and in vivo demonstrated that resistomycin increased the phosphorylation of p38 and MAPKAPK-2 in HepG2 cells. Furthermore, we provided evidence that p38 signaling is involved in resistomycin-induced p38 MAPK pathway effects in HCC, using computer docking models. Our study indicated that resistomycin activates the p38 MAPK signaling pathway by which the growth of HepG2 cells is suppressed for apoptosis and G2/M phase arrest in vitro and in vivo, and it is a promising therapeutic leading compound for drug development in HCC treatment.

Combined use of intravenous and topical tranexamic acid efficiently reduces blood loss in patients aged over 60 operated with a 2-level lumbar fusion.

PURPOSE: The current study was conducted to assess the efficacy and safety of the intravenous (IV) administration combined with topical administration of tranexamic acid (TXA)in patients (aged over 60) scheduled for a 2-level lumbar fusion surgery. METHODS: Two hundred eighty patients scheduled for a 2-level lumbar fusion surgery were randomized into four groups, including an IV group, a local group, a combined group, and a control group. Patients in the combined group, in the IV group, in the topical group, and in the control group were administrated with 15 mg/kg of IV-TXA + 2 g TXA in local, 15 mg/kg IV-TXA, 2 g TXA in local, and 100 ml IV, respectively. The results of total blood loss (TBL), maximum hemoglobin drop, the transfusion rate, and the number of allogeneic blood units were compared. Deep venous thrombosis (DVT) and pulmonary embolism (PE) events were monitored and recorded. RESULTS: The TBL was 635.49 ± 143.60, 892.62 ± 166.85, 901.11 ± 186.25, and 1225.11 ± 186.25 mL for the combined group, the IV group, the topical group, and the control group, respectively (p = 0.015, p = 0.001, respectively). The average maximum hemoglobin drop in the four above groups was 2.18 ± 0.24, 2.80 ± 0.37, 2.40 ± 0.64, and 3.40 ± 1.32 g/dL, respectively. No PE event was reported during the follow-up. Although asymptomatic DVT events were reported by 1, 2, and 2 patients in the combined group, topical group, and control group, respectively, there is no intergroup difference. CONCLUSIONS: The combined use of TXA effectively reduced the total blood loss and blood transfusion rate in patients aged over 60 scheduled for a 2-level lumbar fusion, without increasing the incidence of DVT and PE formation.

[Using of transforaminal lumbar interbody fusion in lumbar revision surgery].

OBJECTIVE: To analyze the effectiveness of transforaminal lumbar interbody fusion (TLIF) for failed back surgery syndrome (FBSS). METHODS: Between October 2003 and December 2007, 36 patients with FBSS were treated with TLIF. There were 19 males and 17 females with an average age of 52.6 years (range, 46-68 years) and an average disease duration of 1.6 years (range, 3 months-15 years). Of 36 patients, reoperation was performed in 25, 10 received 3 operations, and 1 had 5 operations. A total of 50 segments were involved in fusion, including L4,5 in 12 cases, L5, S1 in 10 cases, L3, 4 and L4, 5 double segments in 8 cases, and L4, 5 and L5, S1 double segments in 6 cases. According to X-ray films, CT, and MRI examination, 12 patients were diagnosed as having lumbar instability secondary to total laminectomy, 18 as having recurrence of lumbar disc protrusion, and 6 as having recurrence of lumbar spondylolisthesis. RESULTS: Dural rupture occurred in 1 case and was repaired by suturing without cerebrospinal fluid leakage was observed; 1 had deep incision infection of Staphylococcus; and 1 had transient single irritation sign because of hematoma formation and was cured after symptomatic treatment. The other incisions healed by first intention. No patients had permanent nerve injury or deterioration. Thirty-three cases were followed up 18-72 months (mean, 35.2 months). At 12 months, all the operated segments reached interbody fusion, and no breakage of screw or Cage dislocation occurred. Japanese Orthopaedic Association (JOA) scores showed significant difference (t = 2.45, P = 0.01) between before operation (14.2 +/- 4.1) and 18 months after operation (23.9 +/- 2.6). The rate of clinical improvement was 90.9% (23 cases of excellent, 7 cases of good, 3 cases of acceptable). CONCLUSION: The TLIF simplifies the manipulation of lumbar revision surgery and decreases the operation risk and the operative complications for the treatment of FBSS.

[Anterior cervical discectomy and fusion in the treatment of Hangman's fracture].

OBJECTIVE: To evaluate the efficacies and features of treating Hangman's fracture by anterior cervical discectomy and fusion (ACDF). METHODS: Since June 2005 to December 2008, 36 cases with Hangman's fracture were analyzed with their clinical data including history, symptoms, signs, radiological findings and treatments. According to the classification system designed by Levine and Edwards depending on the radiological manifestations of Hangman's fractures, they were subdivided into type I (n = 9) (conservative therapy), type II (n = 17) and type IIA (n = 10). Conservative therapy was offered to type I in 9 cases, while ACDF with cervical gear protection for 4 weeks after surgery was performed to type II in 17 cases and type IIA in 10 cases. A combination of operation time, days of hospitalization, complications, neurological improvement and fusion rate was assessed. RESULTS: An average follow-up of 15 months (range: 10 - 36) was achieved. No vertebral redisplacement and angulation deformity occurred. Axial pain was relieved in each case. The preoperative neurological deficits in all patients got improvements. No spinal cord injury, vertebral artery injury or cerebrospinal fluid leakage occurred. No complication related to internal fixator was found. Average operative time and hospitalization were similar to those of ACDF for lower cervical spine (98 minutes vs. 9 days). Fusion was achieved within 6 months in all cases. CONCLUSION: ACDF at C2-C3 may be an effective and safe way to treat Hangman's fracture.

[Modified transforaminal lumbar interbody fusion for the treatment of lumbar degenerative disease].

OBJECTIVE: To analyze the clinical effects of modified transforaminal lumbar interbody fusion (TLIF) for the treatment of lumbar degenerative disease. METHODS: From October 2003 to December 2006, 33 patients with lumbar degenerative disease (L3-S1) were treated by modified TLIF. There were 14 males and 19 females with an average age of 52.2 years (33 to 70 years). The median disease course was 1.8 years (4 months to 15 years). A total of 42 levels were fused, including 24 cases of single level and 9 cases of double levels. The results of preoperative diagnosis were lumbar degenerative spondylolisthesis with stenosis (8 cases), isthmic spondylolisthesis (5 cases), degenerative lumbar stenosis (16 cases), huge herniated disc with segmental instability (3 cases) and failed back surgery syndrome (1 case). During the modified TLIF procedure, total inferior facet process and inner half summit of superior facet process of TLIF side were resected to make the posterior wall of foramen opened partly. After the bone graft (3 to 5 mL) was placed into the interbody space, a single rectangle Cage was inserted obliquely from 30 degrees to 40 degrees toward the midline. Combined with pedicle screw instrumentation, TLIF was accomplished. Middle canal and opposite side nerve root decompression were performed simultaneously when necessary. RESULTS: Intraoperative dura mater rupture, postoperative cerebral spinal fluid leakage, deep wound infection and transient nerve root stimulation occurred in 1 case respectively, and were all recovered after treatment. No patients had permanent neurologic deficit or aggravation. All patients were followed up for 20 to 58 months (mean 27.2 months). At the follow-up after 1 year postoperatively, all the operated segments achieved fusion standard and no broken screw or Cage dislocation occurred. All 13 cases of spondylolisthesis were reduced thoroughly and maintained satisfactorily. Nineteen patients remained slight chronic back pain. There was significant difference (P < 0.05) in JOA score between preoperation (14.9 +/- 5.1) and postoperation (25.9 +/- 3.0). The rate of clinical improvement was 80.5% (excellent in 24 cases, good in 7 cases, and fair in 2 cases). CONCLUSION: The modified TLIF carries out the less invasive principles in opening operations, simplifies the manipulation and expands the indication of TLIF to some extent, and the clinical results for the treatment of lumbar degenerative disease is satisfactory.

[Design and experimental study of internal fixator for the reconstruction of lumbar isthmus].

OBJECTIVE: To investigate the clinical applicability and value of internal fixator for the reconstruction of lumbar isthmus in the treatment of lumbar vertebral spondylolysis and to lay a foundation for its clinical application. METHODS: Six teen healthy goats weighing 22.65-31.22 kg were selected to establish the models of vertebral spondylolysis at L5, which thereafter were randomized into two groups (n=8): bone graft group in which 0.8-1.1 g fresh autogenous bone was transplanted into the isthmus spondylolysis area, and internal fixation with bone graft group in which internal fixator was installed before transplanting 0.8-1.1 g fresh autogenous bone into the isthmus spondylolysis area. All animals were killed 8 weeks after operation to receive imaging, topographic anatomy and histology detection. Meanwhile, biomechanics test was performed by using 5 donated vertebral body specimens (4 males and 1 female aged 35-51 years old). The left isthmus of Ls vertebra was transected to serve as lumbar vertebral spondylolysis model. A mini-displacement sensor was put at the transected ends of the isthmus. Then loading was conducted with a constant velocity of 2 mm/min by electronic omnipotent tester simulating the direction of fixation force of the internal fixator, and the deformation value of the transected ends was collected by a dynamic data collector and analyzer. The loading was continued until the vertebra specimens were damaged. The deformation of displacement sensor and the closure of transected ends of the lumbar isthmus were observed. RESULTS: All the goats behaved normally shortly after operation, and no nerve injury induced by operation and no wound infection occurred. Bilaterally oblique X-ray films of lumbar vertebra and topographic anatomy 8 weeks after operation showed the fusion rate of the internal fixation and bone graft group and the bone graft group was 100% and 62.5%, respectively, indicating there was a significant difference (P < 0.05). Histology observation showed 3 goats in the bone graft group presented empty bone trabecula, empty bone lacuna and the disappearance of osteocytes at the transected ends of lumbar isthmus; while in the internal fixation and bone graft group, the bone trabecula grew into cancellous structures with hematopoietic and fatty bone marrow tissue inside, and parts of the bone trabecula had various degrees of mosaic-like pattern. During the upload, the biomechanics test and data processing results showed when the external load was 40 N, the deformation of displacement sensor was identified and the gap between the transected ends of lumbar isthmus started to close; then with the increase of external load, the displacement sensor tended to ascend in a linearity manner; while when the external load was 212 N, the displacement sensor had no further deformation, the gap between the transected ends of lumbar isthmus was completely closed, and the pressor effect appeared. CONCLUSION: The internal fixator for the reconstruction of lumbar isthmus has mechanical effects of stabilizing and elevating pressure with a high fusion rate.

Thoracic ossification of ligamentum flavum caused by skeletal fluorosis.

Thoracic ossification of ligamentum flavum (OLF) caused by skeletal fluorosis is rare. Only six patients had been reported in the English literature. This study reports findings from the first clinical series of this disease. This was a retrospective study of patients with thoracic OLF due to skeletal fluorosis who underwent surgical management at the authors' hospital between 1993 and 2003. Diagnosis of skeletal fluorosis was made based on the epidemic history, clinical symptoms, radiographic findings, and urinalysis. En bloc laminectomy decompression of the involved thoracic levels was performed in all cases. Cervical open door decompression or lumbar laminectomy decompression was performed if relevant stenosis was present. Neurological status was evaluated preoperatively, at the third day postoperatively, and at the end point of follow-up using the Japanese Orthopaedic Association (JOA) scoring system of motor function of the lower extremities. A total of 23 cases were enrolled, 16 (69.6%) males and 7 (30.4%) females, age ranging from 42 to 72 years (mean 54.8 years). All patients came from a high-fluoride area, and 22 (95.7%) had dental fluorosis. Medical imaging showed OLF together with ossification of many ligaments and interosseous membranes, including interosseous membranes of the forearm (18/23 patients 78.3%), leg (14/23 patients 60.9%), and ribs (11/23 patients 47.8%). OLF was classified into five types based on MRI findings: localized (4/23 patients 17.4%), continued (12/23 patients 52.2%), skip (3/23 patients 13.0%), combining with anterior pressure (2/23 patients 8.7%), and combining with cervical and/or lumbar stenosis (2/23 patients, 8.7%). Urinalysis showed a markedly high urinary fluoride level in 14 of 23 patients (60.9%). Patients were followed up for an average duration of 4 years, 5 months. Paired t-test showed that the JOA score was slightly but nonsignificantly increased relative to preoperative measurement 3 days after surgery (P = 0.0829) and significantly increased at the end of follow-up (P = 0.0001). In conclusion, Fluorosis can cause ossification of thoracic ligamentum flavum, as well as other ligaments. Comparing with other OLF series, a larger number of spinal segments were involved. The diagnosis of skeletal fluorosis was made by the epidemic history, clinical symptom, imaging study findings, and urinalysis. En bloc laminectomy decompression was an effective method.