Objective To investigate the effects of R-spondin 2 (Rspo2) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and bone mineral content in ovariectomized mice. Methods BMSCs were extracted from the bone marrow of the long bones of 7 4-week-old female C57BL/6 mice using whole bone marrow culture and passaged. After the cell phenotype was identified by flow cytometry, the 3rd generation cells were co-cultured with 10, 20, 40, 80, and 100 nmol/L Rspo2. Then, the cell activity and proliferative capacity were determined by cell counting kit 8 (CCK-8), and the intervention concentration of Rspo2 was screened for the subsequent experiments. The osteogenic differentiation ability of BMSCs was detected by alkaline phosphatase (ALP) staining, and the mRNA levels of osteogenesis-related genes [RUNX family transcription factor 2 (Runx2), collagen type Ⅰ alpha 1 (Col1), osteocalcin (OCN)] were detected by real-time fluorescence quantitative PCR (RT-qPCR). In addition, 18 10-week-old female C57BL/6 mice were randomly divided into sham operation group (sham group), ovariectomy group (OVX group), and OVX+Rspo2-intervention group (OVX+Rspo2 group), with 6 mice in each group. The sham group only underwent bilateral back incision and suturing, while the other two groups established osteoporosis mouse models by bilateral ovarian castration. Then, the mice were given a weekly intraperitoneal Rspo2 (1 mg/kg) treatment in OVX+Rspo2 group and saline at the same dosage in sham group and OVX group. After 12 weeks of treatment, the body mass and uterus mass of the mice were weighed in the 3 groups to assess whether the OVX model was successfully prepared; the tibia bones were stained with HE and immunohistochemistry staining to observe the changes in tibial bone mass and the expression level of Runx2 protein in the bone tissues. Blood was collected to detect the expressions of bone metabolism markers [ALP, OCN, type Ⅰ procollagen amino-terminal peptide (PINP)] and bone resorption marker [β-collagen degradation product (β-CTX)] by ELISA assay. Micro-CT was used to detect the bone microstructure changes in the tibia, and three-dimensional histomorphometric analyses were performed to analyze the trabeculae thickness (Tb.Th), trabeculae number (Tb.N), trabeculae separation (Tb.Sp), and bone volume fraction (BV/TV). Results CCK-8 assay showed that Rspo2 concentrations below 80 nmol/L were not cytotoxic (P>0.05), and the cell viability of 20 nmol/L Rspo2 group was significantly higher than that of the control group (P<0.05). Based on the above results, 10, 20, and 40 nmol/L Rspo2 were selected for subsequent experiments. ALP staining showed that the positive cell area of each concentration of Rspo2 group was significantly larger than that of the control group (P<0.05), with the highest showed in the 20 nmol/L Rspo2 group. The expression levels of the osteogenesis-related genes (Runx2, Col1, OCN) significantly increased, and the differences were significant between Rspo2 groups and control group (P<0.05) except for Runx2 in the 40 nmol/L Rspo2 group. In animal experiments, all groups of mice survived until the completion of the experiment, and the results of the body mass and uterus mass after 12 weeks of treatment showed that the OVX model was successfully prepared. Histological and immunohistochemical staining showed that the sparseness and connectivity of bone trabecula and the expression of Runx2 in the OVX group were lower than those in the sham group, whereas they were reversed in the OVX+Rspo2 group after treatment with Rspo2, and the differences were significant (P<0.05). ELISA assay showed that compared with the sham group, the serum bone metabolism markers in OVX group had an increase in ALP and a decrease in PINP (P<0.05). After Rspo2 intervention, PINP expression significantly reversed and increased, with significant differences compared to the sham group and OVX group (P<0.05). The bone resorption marker (β-CTX) was significantly higher in the OVX group than in the sham group (P<0.05), and it was significantly decreased in the OVX+Rspo2 group when compared with the OVX group (P<0.05). Compared with the sham group, Tb.Th, Tb.N, and BV/TV significantly decreased in the OVX group, while Tb.Sp significantly increased (P<0.05); after Rspo2 intervention, all of the above indexes significantly improved in the OVX+Rspo2 group (P<0.05) except Tb.Th. Conclusion Rspo2 promotes differentiation of BMSCs to osteoblasts, ameliorates osteoporosis due to estrogen deficiency, and promotes bone formation in mice.
Objective To explore the surgical procedure and effectiveness of total spondylectomy and spine reconstruction through posterior approach to treat thoracolumbar vertebra tumor. Methods Between June 2004 and July 2008, 14 cases of thoracolumbar vertebra tumor underwent one-stage total spondylectomy through posterior approach and spine reconstruction with posterior pedicle screw system and bone graft. There were 11 males and 3 females with a mean age of 47.2 years (range, 36-60 years). The disease duration was 3-15 months. Affected segments included T3 in 1 case, T4 in 3 cases, T8 in 3 cases, T9 in 2 cases, T10 in 3 cases, T12 in 1 case, and L1 in 1 case. The postoperative pathological results were 3 cases of bony giant cell tumor, 1 case of osteoblastoma, 2 cases of osteosarcoma, and 8 cases of metastatic tumor. According to Tomita et al. grading system, there were 1 case of type II, 5 cases of type III, 3 cases of type IV, and 5 cases of type V. According to Frankel classification of preoperative spinal cord function, 3 cases were rated as grade B, 4 as grade C, 5 as grade D, and 2 as grade E. Results Wound heal ing by first intention was obtained in all cases, and no blood vessel and nerve injury occurred. Fourteen patients were followed up 11-64 months (mean, 32.5 months). The local pain was rel ieved significantly. At 6-8 months after operation, the X-ray films and CT showed that bone graft fusion at Bridwell I grade was achieved. At 10 months, the postoperative spinal cord function was improved from grade B to grade D in 2 cases, from grade C to grade D in 1 case, and the other 9 cases reached grade E. The patients had normal walking function. Two patients died of l iver metastasis and brain metastasis at 11 and 15 months postoperatively, respectively; 1 patient with osteoscarcoma died of lung metastasis at 16 months; and 1 case of osteoscarcoma developed local recurrence at 8 months postoperatively. Internal fixation was rel iable without loosening and breakage and the spine was stable. Conclusion Total spondylectomy and spine reconstruction through posterior approach is an effective method with advantages of relative minimal injury, radical tumor excision, low local recurrence, and adequate spinal cord decompression.
ObjectiveTo investigate the feasibility of one-stage total spondylectomy by anterior and posterior approaches for treating solitary plasmacytoma (SP) of cervical spine.MethodsBetween June 2000 and March 2015, the clinical data, diagnosis, and treatment of 12 patients with SP of cervical spine were retrospectively analyzed. There were 8 males and 4 females, with a mean age of 46 years (range, 28-74 years). The mean disease duration was 8.4 months (range, 3-15 months). There were 2 cases in C2 segment, 4 cases in C3, 2 cases in C4, 1 recurrent case in C5, C6, and 3 cases in C7. According to WBB (Weinstein-Boriani-Biagini) surgical staging system, the tumor mainly occupied the vertebral body areas and expanded outside to 4 or 9 radiating zone in 8 cases, expanded both outsides to 4 or 9 radiating zones in 4 cases; 7 cases of them affected A to D layers, other 5 cases affected B to D layers. Preoperative Frankel system showed grade B in 1 case, grade C in 6 cases, and grade D in 5 cases. One vertebra segment was involved in 11 cases, and 2 vertebra segments were involved in 1 case. The unilateral vertebral artery was involved in 4 cases and bilateral vertebral artery in 1 case. One-stage total spondylectomy by anterior and posterior approaches for SP of cervical spine was applied in all patients, which all received adjuvant radiotherapy postoperatively. The operation time, intraoperative blood loss, complications, Frankel system, and visual analogue scale (VAS) score were recorded and analyzed.ResultsThe operation time was 6.8-9.3 hours (mean, 7.2 hours), and the intraoperative blood loss was 1 100-2 600 mL (mean, 1 600 mL). Esophageal leakage occurred in 1 patient with C5, C6 recurrent SP at 1 week after operation and 1 case presented with fat liquefaction of wound, who were cured by symptomatic treatment. Other patients showed no neurological deficit, major vascular injury (especially vertebral artery), phrenic nerve injury, superior laryngeal nerve injury, laryngeal nerve injury, respiratory failure, or other complications. All the 12 patients were followed up 27-98 months (mean, 58 months). The symptoms of spinal cord compression disappeared or improved after operation. At last follow-up, the nerve function was recovered to Frankel grade E in all patients; the VAS score decrease to 1.1±0.7, showing significant difference when compared with preoperative value (6.7±2.7) (t=2.485, P=0.014). Two patients had local recurrence at 29 months and 37 months after operation respectively, which were treated with adjuvant chemotherapy. One of them finally progressed to multiple myeloma and died of multiple organ failure after 43 months, the other one survived with residual tumor. One case presented with internal fixator loosening and breaking at 4 years after operation, who was performed revision surgery. The other patients had no tumor recurrence or malignant change during the follow-up, no complication such as internal fixator loosening or breaking occurred.ConclusionFor patients with SP of cervical spine, surgical intervention is an acceptable treatment option. One-stage total spondylectomy by anterior and posterior approaches can decline the local recurrence and relieve the symptoms of spinal cord compression, so as to improve patients’ life quality. However, patients with progression to multiple myeloma should be treated with individualized therapeutic regimen, and the prognosis may be poor.
Objective To investigate the clinical therapeutic effects on malignant spinal tumors treated by percutaneous vertebroplasty(PVP) under the guidance of the digital subtraction angiography(DSA). Methods A retrospective analysis was performed in 196 patients (99 males and 97 females, aged 23-85 years, averaged 60.4 years) with malignant spinal tumors, who underwent the PVP treatment combined with standard chemotherapy and other comprehensive treatment from January 2002 to January 2005. The malignant spinal tumors had their origins as follows: lung cancer (66 cases), breast cancer (55 cases), liver cancer (19 cases), colon cancer (15 cases), stomach cancer (9 cases), prostate cancer (12 cases), multiple myeloma (16 cases), and malignant lymphoma of the spine (4 cases).The metastatic tumors involved the cervical vertebra (32 cases), thoracic vertebra (93 cases), lumbar vertebra (71 cases), and spinal column, including 1 vertebral segment (135 cases), 2 segments (50 cases), and more than 3 segments (11 cases). During the follow-up survey, changes in the visual analogue pain scale(VAS) and changes in the X-ray measurements of the average anterior height, midline height, and posterior height of the diseased vertebra were observed. Results The follow-up for 6 months to 3 years revealed that the percutaneous vertebroplasty on279 vertebral segments had a success with an operational success rate of 100%. Bone cement was injected into the lesions 1-9 ml per segment of the spine. The postoperative X-ray and CT evaluations revealed that spinal stabilization was obtained in all the patients. After operation, 193（98.5%）patients had an obvious decrease or disappearance of the pain in the lower back, and only 3 (1.5%) patients had no obvious improvement in the pain. There was a significant statist-ical difference in the VAS scores between before operation and after operation (Plt;0.05). There were also significant statistical differences in the average anterior height of the diseased vertebra between before operation and after operation（15.71±2.80 mm vs 16.61±3.01 mm）, in the midline height（13.65±2.93 mm vs 14.52±2.72 mm）, and in the posterior height（23.67±2.81 mm vs 23.70±3.13 mm,Plt;0.05). The patients with lung or liver cancer had a mean survival time of 9 monthsafter PVP; the patients with breast cancer, stomach cancer, prostate cancer, lymphoma, or other metastatic tumors had a mean survival time of 18 months. The patients with multiple myeloma had a mean survival time of 27 months. The differences were statistically different (Plt;0.01). Conclusion PVP under the guidance of the DSA is an easier operation witha small wound and few complications. It can effectively alleviate the patient’s pain due to metastatic spinal tumor, stabilize the spine, improve the patient’s quality of life, and reduce the incidence of paraplegia.
Objective To observe the effects of Crumbs (Crb) proteins on different types of zebrafish photoreceptors. Methods The retinal cell population dynamics of adult wild-type zebrafish and Tg (RH2-2:Crb2b-sf-EX/RH2-2:GFP)pt108b transgenic zebrafish (called pt108b zebrafish for short) were evaluated by monitoring the densities of three categories of retinal photoreceptors (rod cells, UV cone cells and RGB cone cells) in different retinal regions, which were visualized by Feulgen nuclear staining histology technique. Results The wild-type zebrafish retinal photoreceptor cell densities are generally higher in the central region than the peripheral regions. Compared with wild-type zebrafish, pt108b zebrafish had much less RGB cone cells at the top of outer nuclear layer, and no RGB cone cells at the central and intermediate regions of retina. While pt108b zebrafish had normal density of UV cone cells at the top of rods and the bottom of outer limiting membrane, they had much higher density of rods. Conclusions Crb proteins may affect the zebrafish retinal cell densities of different photoreceptor types.
【Abstract】 Objective To investigate the clinical significances of the thoracic pedicle classification determined by inner cortical width of pedicle in posterior vertebral column resection (PVCR) with free hand technique for the treatment of rigid and severe spinal deformities. Methods Between October 2004 and July 2010, 56 patients with rigid and severe spinal deformities underwent PVCR. A total of 1 098 screws were inserted into thoracic pedicles at T2-12. The inner cortical width of the thoracic pedicle was measured and divided into 4 groups: group 1 (0-1.0 mm), group 2 (1.1-2.0 mm), group 3 (2.1-3.0 mm), and group 4 (gt; 3.1 mm). The success rate of screw-insertion into the thoracic pedicles was analyzed statistically. A new 3 groups was divided according to the statistical results and the success rate of screw-insertion into the thoracic pedicles was analyzed statistically again. And statistical analysis was performed between different types of thoracic pedicles classification for pedicle morphological method by Lenke. Results There were significant differences in the success rate of screw-insertion between the other groups (P lt; 0.008) except between group 3 and group 4 (χ2=2.540，P=0.111). The success rates of screw-insertion were 35.05% in group 1, 65.34% in group 2, and 88.32% in group 3, showing significant differences among 3 groups (P lt; 0.017). According to Lenke classification, the success rates of screw-insertion were 82.31% in type A, 83.40% in type B, 80.00% in type C, and 30.28% in type D, showing no significant differences (P gt; 0.008) among types A, B, and C except between type D and other 3 types (P lt; 0.008). In the present study, regarding the distribution of different types of thoracic pedicles, types I, II a, and II b thoracic pedicles accounted for 17.67%, 16.03%, and 66.30% of the total thoracic pedicles, respectively. The type I, II a, and II b thoracicpedicles at the concave side accounted for 24.59%, 21.13%, and 54.28%, and at the convex side accounted for 10.75%, 10.93%, and 78.32%, respectively. Conclusion A quantification classification standard of thoracic pedicles is presented according to the inner cortical width of the pedicle on CT imaging: type I thoracic pedicle, an absent channel with an inner cortical width of 0-1.0 mm; type II thoracic pedicle, a channel, including type IIa thoracic pedicle with an inner cortical width of 1.1-2.0 mm, and type IIb thoracic pedicle with an inner cortical width more than 2.1 mm. The thoracic pedicle classification method has high prediction accuracy of screw-insertion when PVCR is performed.
Lumbar spondylolisthesis is a common condition in spinal surgery, which is often characterized by lower back and leg pain and numbness. There are various treatment methods for this condition, and different treatment plans should be adopted according to different situations. Traditional open surgery methods are relatively traumatic and have longer recovery times, while minimally invasive spine techniques have advantages such as smaller incisions, less bleeding, higher fusion rates, and faster recovery. This review summarizes the relevant literature on the application of minimally invasive techniques in the treatment of lumbar spondylolisthesis in recent years, analyzes and compares the advantages and disadvantages of different approaches and endoscopic techniques, as well as reduction, decompression, and fusion effects. The aim is to provide reference for surgeons in selecting surgical procedures for the treatment of lumbar spondylolisthesis.
【摘 要】 目的 探討布魯桿菌病性脊椎炎有效的外科治療方法及臨床效果。 方法 2002年1月－2010年10月，對78例具有手術指征的胸腰椎布魯桿菌病性脊椎炎患者采用一期病灶清除聯合后路椎弓根內固定治療。男42例，女36例；年齡24～65歲，平均45歲。病程8～29個月，平均12個月。2個椎體受累70例，3個椎體受累8例。均合并不同程度神經功能損傷。術后隨訪行疼痛視覺模擬評分（VAS）、神經功能Frankel分級、臨床療效評價以及X線片、MRI影像學觀察。 結果 術后78例均獲隨訪，隨訪時間12～30個月，平均26個月。無竇道形成及復發。術前及術后1、3、6、12個月VAS評分分別為（9.2 ± 0.6）、（2.4 ± 0.3）、（1.0 ± 0.2）、（0.5 ± 0.4）及0分，術后各時間點均較術前顯著改善（P lt; 0.05）。術后各時間點神經功能Frankel分級均較術前顯著改善，差異均有統計學意義（P lt; 0.05），其中C、D級改善最為顯著。臨床療效評價：術后12個月內均無加重患者；隨時間推移，改善及無變化患者逐漸轉向痊愈，術后12個月改善率和痊愈率分別為100%和91.03%。影像學評價：術前及術后1、3、6、12個月影像學盲測評價指標評分分別為（0.17 ± 0.03）、（4.11 ± 0.09）、（4.68 ± 0.04）、（4.92 ± 0.08）及5分，術后各時間點均較術前顯著改善（P lt; 0.05）。 結論 一期病灶清除聯合后路椎弓根內固定治療胸腰椎布魯桿菌病性脊椎炎，在解除疼痛、穩定脊柱、恢復神經功能及早期康復方面均有明顯優勢。
ObjectiveTo explore an method and the effectiveness of surgical treatment of thoracic and lumbar tuberculosis complicated with severe kyphotic deformity (Cobb angle≥55°) and paraplegia. MethodsBetween January 2009 and January 2013, 13 cases of thoracic and lumbar tuberculosis complicated with severe kyphotic deformity and paraplegia were treated by one-stage posterior vertebral column resection (PVCR), debridement, bone grafting, and instrumentation fixation. Of 13 patients, 7 were male and 6 were female with an average age of 23.5 years (range, 14-49 years). The disease duration was 13-38 months (mean, 19 months). The Cobb angle of kyphosis was (65.23±7.95)°. The visual analogue scale score (VAS) was 7.38±0.31. In 13 patients with incomplete paraplegia, 1 case was classified as Frankel grade B, 7 cases as grade C, and 5 cases as grade D. The lesion involved 2 vertebrae bodies in 7 cases (T8, 9 in 1 case, T11, 12 in 2 cases, and T12, L1 in 4 cases), 3 vertebrae bodies in 4 cases (T10-12 in 2 cases, T9-11 in 1 case, and T11-L1 in 1 case), and 4 vertebrae bodies in 2 cases (T4-7 in 1 case and T6-9 in 1 case). Imaging examination showed paravertebral abscess in 10 cases. ResultsHealing of incision by first intention was obtained in all patients. The neurological injury and pulmonary infection occurred in 3 cases and 2 cases respectively, which were cured after symptomatic treatment. Thirteen patients were followed up 12-48 months (mean, 17 months). The erythrocyte sedimentation rate restored to normal level in all cases at 3-7 months after operation. All the patients achieved bony fusion at 10-20 months (mean, 14 months) after operation. No fixation loosening, displacement, or fracture occurred during follow-up. Common toxic symptom of tuberculosis disappeared, and there was no recurrence of local tuberculosis. The Cobb angle of kyphosis was corrected to (22.38±1.76)° at 1 week and (22.15±1.83)° at last follow-up, showing significant difference when compared with preoperative one (P < 0.05). There was no significant difference in Cobb angle of kyphosis between at 1 week after operation and at last follow-up (P > 0.05). The Frankel grading was grade B in 1 case, grade C in 10 cases, and grade D in 2 cases at 1 week after operation; and it was grade D in 1 case and grade E in 12 cases at last follow-up; significant differences were found between at pre-and post-operation (P < 0.05), and between at 1 week after operation and at last follow-up (P < 0.05). The VAS score was 4.08±0.76 at 1 week and 0.62±0.14 at last follow-up, showing significant differences between at pre-and post-operation (P < 0.05) and between at 1 week after operation and at last follow-up (P < 0.05). ConclusionOne-stage PVCR, debridement, bone grafting, and instrumentation fixation is proved to be successful in treating thoracic and lumbar tuberculosis complicated with severe kyphotic deformity and paraplegia.