[Effects of plumbagin on proliferation and apoptosis of human hepatocellular carcinoma cells based on CKB/p53 signaling pathway].

To investigate the effects of plumbagin on the proliferation and apoptosis of human hepatoma Huh-7 cells and its mechanism based on the creatine kinase B(CKB)/p53 signaling pathway. Huh-7 cells were treated with plumbagin from 1 to 12 µmol·L~(-1) for cell counting kit-8(CCK-8) assay, and 1, 3, and 6 µmol·L~(-1) were determined as low, medium, and high concentrations of plumbagin for subsequent experiments. CKB gene was knocked out in Huh-7 cells by clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated proteins(Cas)-9 gene editing technology. CKB overexpression lentivirus was transfected into Huh-7 cells to up-regulate the expression of CKB. Cell proliferation and apoptosis were detected by plate cloning assay and flow cytometry. The mRNA expression of CKB was detected by quantitative real-time PCR(qRT-PCR). CKB, p53, mouse double minute 2 homolog(MDM2), B-cell lymphoma 2(Bcl-2), Bcl-2 associated X protein(Bax), and caspase-3 protein were detected by Western blot(WB). The results showed that plumbagin significantly inhibited the proliferation of Huh-7 cells and induced cell apoptosis. Compared with the control group, the apoptosis level was significantly increased in the plumbagin group, while the apoptosis level was significantly decreased in the plumbagin combined with the apoptosis inhibitor group. Plumbagin significantly down-regulated the protein expression levels of CKB, Bcl-2, and MDM2 and up-regulated the protein expression levels of p53, Bax, and caspase-3. Knockdown of the CKB gene decreased the proliferative ability of Huh-7 cells, increased the apoptotic rate, decreased the expression levels of Bcl-2 and MDM2 proteins, and increased the expression levels of p53, Bax, and caspase-3 proteins. After up-regulation of CKB expression, the proliferation ability of Huh-7 cells was enhanced, and the protein expression levels of Bcl-2 and MDM2 were elevated. The protein expression levels of p53, Bax, and caspase-3 were decreased. In addition, plumbagin reversed the effect of overexpression of CKB on the proliferation and apoptosis of Huh-7 cells. In conclusion, plumbagin significantly inhibited the proliferative ability of Huh-7 cells, and the mechanism may be related to the inhibition of CKB expression, activation of the p53 signaling pathway, and regulation of the expression of mitochondrial-associated apoptotic proteins, ultimately inducing cell apoptosis.

The characteristic and prognostic role of blood inflammatory markers in patients with Huntington's disease from China.

Objectives: This study aims to elucidate the role of peripheral inflammation in Huntington's disease (HD) by examining the correlation of peripheral inflammatory markers with clinical manifestations and disease prognosis. Methods: This investigation involved 92 HD patients and 92 matched healthy controls (HCs). We quantified various peripheral inflammatory markers and calculated their derived metrics including neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), and systemic immune-inflammation index (SII). Clinical assessments spanning cognitive, motor, and disease severity were administered. Comparative analysis of inflammatory markers and clinical correlations between HD and controls was performed. Kaplan-Meier survival analysis and Cox regression model were used to assess the effect of inflammatory markers on survival. Results: The study revealed that HD patients had significantly reduced lymphocyte counts, and LMR. Conversely, NLR, PLR, and SII were elevated compared to HCs. Lymphocyte levels inversely correlated with the age of onset and monocyte levels inversely correlated with the UHDRS-total functional capacity (TFC) scores. After adjusting for age, sex, and CAG repeat length, lymphocyte count, NLR, PLR, and SII were significantly correlated with the progression rate of TFC scores. Elevated levels of white blood cells and monocytes were associated with an increased risk of disability and mortality in the HD cohort. Conclusion: Our findings indicate that HD patients display a distinct peripheral inflammatory profile with increased NLR, PLR, and SII levels compared to HCs. The peripheral inflammation appears to be linked with accelerated disease progression and decreased survival in HD.

The protocol for the Prospective Urban Rural Epidemiology China Action on Spine and Hip status study.

The Prospective Urban Rural Epidemiology (PURE) China Action on Spine and Hip status (CASH) study focused on the prevalence of osteoporosis and spinal fracture in China. The aim of the PURE CASH study is to determine the prevalence of osteoporosis and spinal fracture, and explore the potential relationship between spinal fracture and bone mineral density (BMD). This study is a prospective large-scale population study with a community-based sampling and recruitment strategy. The aim is to determine the prevalence of osteoporosis and vertebral fracture in this population, to evaluate the association between vertebral fractures and BMD values, and to assess the prediction power of BMD for incident fractures. Participants in the PURE CASH study are all from the PURE study in China, recruited from 12 centers in 7 Chinese provinces. The inclusion criteria are that participants should be aged more than 40 years and able to give informed consent. Exclusion criteria are pregnant women, individuals with metal implants in the lumbar spine, use of medications or the existence of any disease or condition known to have a major influence on BMD, and inability to give informed consent. A total of 3,457 participants undergo a quantitative computed tomography (QCT) scan of the upper abdomen. The scanning parameters are as follows: 120 kVp at all centers, mAs between 75 and 200, FOV 40 cm×40 cm. The BMD values of L1 to L3 are measured, and the average BMD calculated. The American College of Radiology QCT criteria for the diagnosis of osteoporosis is applied to determine the presence of osteoporosis. The scout view images of T4-L4 vertebrae are reviewed by two experienced radiologists for semi-quantification of vertebral fractures according to Genant's method.

Anterior and Posterior Instrumentation with Different Debridement and Grafting Procedures for Multi-Level Contiguous Thoracic Spinal Tuberculosis.

OBJECTIVE: To evaluate the clinical outcomes of anterior and posterior instrumentation with different debridement and graft fusion methods for multi-level contiguous thoracic spinal tuberculosis. METHODS: We retrospectively evaluated 81 patients with multi-level contiguous thoracic spinal tuberculosis who underwent anterior or posterior instrumentation combined with different methods of debridement, decompression, and graft fusion from January 2002 to December 2012. All patients were divided into an anterior instrumentation group and a posterior instrumentation group. In the anterior instrumentation group, there were 39 patients who underwent transthoracic debridement. In the posterior instrumentation group, there were 34 patients who underwent trans-costotransverse decompression and strut grafting with posterior instrumentation, and another 8 patients underwent combined anterior debridement and strut grafting with posterior instrumentation in a single-stage or two-stage procedure. The kyphotic angles were calculated from lateral spinal X-rays using the modified Konstam method. The symptoms and signs of tuberculosis, fusion level, fusion time of the bone graft, average kyphosis angle, average correction, average loss of correction, and clinical complications were recorded. The average follow-up period was 37 months (range, 17-72 months). RESULTS: The cohort consisted of 47 males and 34 females with an average age of 38 years. The mean durations of the operations were 3.5 ± 0.4 h in the anterior group and 4.0 ± 0.3 h in the posterior group ( P < 0.05). The mean blood loss volumes during surgery were 450 ± 42 and 560 ± 51 mL for the anterior group and the posterior group, respectively ( P < 0.01). The kyphotic deformities were corrected from 32.1° ± 10.3° to 10.2° ± 2.1° in the anterior group and from 33.8° ± 11.7° to 12.6° ± 2.7° in the posterior group ( P < 0.01). The neurologic statuses of the 23 patients with preoperative neurologic deficits improved in each group. Fusion was confirmed radiographically at 5.4 ± 1.2 months (range, 4-12 months) in the anterior group and 5.6 ± 1.4 months (range, 4-13 months) in the posterior group ( P > 0.05). Postoperative relapses were noted in 1 and 3 patients in the anterior and the posterior group, respectively. CONCLUSION: Posterior instrumentation was more effective than anterior instrumentation in the correction of kyphosis and the maintenance of the correction. However, postoperative sinus formation was more frequent in patients who underwent a single-stage posterior procedure.

[Case-control studies of two kinds of method for the treatment of lumbar tuberculosis with psoas abscess].

OBJECTIVE: To compare two kinds of method for treating lumbar tuberculosis with psoas abscess, to provide reference for clinical reasonable select of therapy treatment. METHODS: From January 2010 to January 2013,42 patients with lumbar tuberculosis combined with psoas abscess with obvious surgical indications were enrolled, including 24 males and 18 females with an average age of (38.5 ± 10.2) years old ranging from 21 to 63 years old. All patients were followed up for 18 to 24 months with an average of 20.9 months. Twenty-two patients underwent posterior vertebral body lesions cleared, bone graft fusion and internal fixation and percutaneous puncture catheter drainage for treatment of psoas major abscess as group A, and twenty patients underwent one-stage extraperitoneal approach to remove abscess, posterior vertebral body lesions cleared, bone graft fusion and internal fixation as group B. The operative time, loss of blood, length of hospital stay, clinical cure rate and other clinical results for the two groups were analyzed and compared. RESULTS: The loss of blood was (452.3 ± 137.6) ml in group A and (603.5 ± 99.6) ml in group B, there was significant statistical difference (P < 0.05). The time of operation was (193.6 ± 91.2) min in group A and (230.5 ± 56.6) min in group B, there was significant statistical difference (P < 0.05). The time of operation and the loss of blood in group A were obviously less than which in group B. In group A 20 cases were cured and 2 cases relapsed, 19 cases were cured and 1 case relapsed in group B, there was no significant statistical differences between two groups regarding cure rate with chi-square test (χ² = 0.000, P = 1.000). All patients in two groups obtained good clinical curative effect. There were no significant statistical difference between two groups regarding for length of hospital stay with t-test (P > 0.05). CONCLUSION: Lumbar spinal tuberculosis with psoas abscess is not absolute indications for anterior open operation. Compared with the combined anterior and posterior surgical procedure, the percutaneous puncture catheter drainage combined with posterior debridement, interbody fusion and internal fixation can achieve the same clinical effect but less trauma for the patients.

The classification of recurrent spinal epidural hematoma: a review of the literature and a comparison with the cases.

Symptomatic postoperative spinal epidural hematoma (SEH) and spontaneous spinal epidural hematoma (SSEH) are both rare conditions, and recurrent SEH occurs even less frequently. Therefore, we describe a case of symptomatic postoperative SEH after surgical evacuation of SSEH, which was diagnosed using magnetic resonance imaging (MRI) and managed with negative pressure wound therapy (NPWT). The authors classified the reported recurrent SEHs into two types based on the cause of their previous hematoma, which can be classified as spontaneous or postoperative. The characteristics, diagnosis, managements, and results of recurrent SEHs were analyzed. The authors suggest that the postoperative SEH in the Type II will be treated with NPWT, and the new classification will be helpful for prognosis, diagnosis, and management of the recurrent SEHs.

[The cost effectiveness analysis of minimally invasive surgery and conservative treatment in elderly osteoporotic spinal fracture].

OBJECTIVE: To evaluate the cost effectiveness of conservative treatment, percutaneous vertebroplasty(PVP)and percutaneous kyphoplasty(PKP)for elderly osteoporotic vertebral compression fracture(OVCF). METHODS: The clinical data of 152 patients with osteoporotic vertebral compression fractures, collected in the orthopedic department of 309th Hospital of PLA from October 2013 to July 2014, was retrospectively analyzed. According to the therapeutic methods, the patients who met the inclusion criteria were divided into conservative treatment group (51 cases), percutaneous vertebroplasty group (50 cases) and percutaneous kyphoplasty group(51 cases). The average medical cost (C) in hospital period and 1 year after discharging, and the treatment effect (E) according to standard of "cure" (VAS score less than or equal to 2) or "improvement" (VAS score was 3 to 8) was recorded. Then the C/E value indicated the cost effectiveness in different standards. RESULTS: The average hospitalization days of the PVP and PKP group was 3 to 5 days with an average of(3.4±0.6) days. The conservative group was 12 to 15 days with an average of (14.0±0.6) days. During the hospitalization period, the cost effectiveness of the conservative group, PVP group and PKP group were RMB 1 253.88, 935.75, 983.99 yuan, respectively, according to the standard of "cure". The PVP group was superior to the PKP group and the latter was superior to the conservative group. If "improvement" was used as the standard of evaluation, the results were RMB 97.80, 449.16, 501.84 yuan, respectively, suggesting that the conservative group was better than the PVP group and the latter was better than the PKP group. After hospital discharge, the cost effectiveness of the conservative group, PVP group and PKP group were RMB 3 834.05, 1 878.41 and 1 916.11 yuan, respectively, according to the standard of "cure". The PVP group was superior to the PKP group and the latter exceeded the conservative group. CONCLUSIONS: The study showed that the PVP was the best choice at the evaluation criterion of "cure", while taking "improvement" as the evaluation criterion, the conservative treatment was the best one. Either way, the PVP was the best choice after hospital discharge.

The PI3K/AKT cell signaling pathway is involved in regulation of osteoporosis.

BACKGROUND: Osteoporosis is a systemic skeletal disease with the high incidence, serious complications, financial burden, and heavily decrease in living quality. METHODS: Proliferation of osteoblast was tested by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method, alkaline phosphatase (ALP) activity of osteoblasts was tested by ALP REAGENT, Calcium level was determined by a colorimetric assay, mRNA expression of phosphoinositide-3 kinase (PI3K), 3-phosphoinositide-dependent protein kinase 1 (PDK1), Akt, Caspase-3, Caspase-7, Caspase-9, osteocalcin (OCN), Osterix and Runx2 of osteoblasts was tested by RNA preparation and quantitative reverse transcription polymerase chain reaction (RT-PCR), and protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt was measured by Western Blot analysis. RESULTS: In osteoporosis model rats, it found that mRNA expression of PI3K, PDK1 and Akt showed no changes while protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt in bone tissue was decreased dramatically. To further characterize the molecular mechanisms that regulate osteoporosis, we examined the contribution of the PI3K/Akt cell signaling pathway in cultured osteoblasts. It suggested that, the blockade of PI3K activation by LY294002, a specific inhibitor of the PI3K/Akt signaling pathway in osteoblasts, heavily inhibited cell proliferation, ALP activity, calcium accumulation, and mRNA expression of OCN, Osterix and Runx2. However, mRNA expression of Caspase-3 and Caspase-9 was promoted accordingly. CONCLUSION: The in vivo and in vitro studies indicated that the PI3K/Akt cell signaling pathway is involved in the inhibition of osteoporosis through promoting osteoblast proliferation, differentiation and bone formation.

MicroRNA-146a reduces IL-1 dependent inflammatory responses in the intervertebral disc.

Because miR-146a expression in articular chondrocytes is associated with osteoarthritis (OA), we assessed whether miR-146a is linked to cartilage degeneration in the spine. Monolayer cultures of nucleus pulposus (NP) cells from the intervertebral discs (IVD) of bovine tails were transfected with a miR-146a mimic. To provoke inflammatory responses and catabolic extracellular matrix (ECM) degradation, cells were co-treated with interleukin-1 (IL-1). Transfection of miR-146a decreases IL-1 induced mRNA levels of inflammatory genes and catabolic proteases in NP cells based on quantitative real-time reverse transcriptase PCR (qRT-PCR) analysis. Similarly, miR146a suppresses IL-1 induced protein levels of matrix metalloproteinases and aggrecanases as revealed by immunoblotting. Disc segments from wild type (WT) and miR-146a knockout (KO) mice were cultured ex vivo in the presence or absence of IL-1 for 3days. Histological and immuno-histochemical (IHC) analyses of disc organ cultures revealed that IL-1 mediates changes in proteoglycan (PG) content and in-situ levels of catabolic proteins (MMP-13 and ADAMTS-5) in the nucleus pulposus of the disc. However, these IL-1 effects are more pronounced in miR-146a KO discs compared to WT discs. For example, absence of miR-146a increases the percentage of MMP-13 and ADAMTS-5 positive cells after treatment with IL-1. Thus, miR-146a appears to protect against IL-1 induced IVD degeneration and inflammation. Stimulation of endogenous miR-146a expression or exogenous delivery of miRNA-146a are viable therapeutic strategies that may decelerate disc degeneration and regain a normal homeostatic balance in extracellular matrix production and turn-over.

[Analysis of clinical characteristics of elderly patients with spinal tuberculosis and its clinical effects with conservative treatment].

OBJECTIVE: To analyze the clinical characteristics of elderly patients with spinal tuberculosis and explore its clinical effects with anti-TB drugs alone. METHODS: From January 2008 to July 2010, the data of 36 patients with spinal tuberculosis underwent conservative treatment of anti-TB drugs alone were analyzed. There were 19 males and 17 females with an average age of 73.5 years (ranged, 60 to 85). All patients were in the active phase with high ESR and CRP levels and were treated with 3HRZE/6-9HRE (course from 9 to 12 months). According to clinical symptoms, chemical examination, radiological image to adjust drug and depending on VAS score to evaluate pain. RESULTS: All the patients were followed up from 8 to 24 months with an average of 15 months. Tuberculose of 31 patients healed after chemotherapy from 9 to 12 months and ESR and CRP recovered normally. Levofloxacin and para-amino salicylic acid were used in 4 cases because of 4 cases occurred drug fast for RFP or INH, after 15 months, their obtained healing. Symptom of 1 case got worse during chemotherapy, and surgical treatment were performed, after 3 months, ESR and CRP recovered normally, X-ray and CT showed spinal osteosclerosis and fusion without significant kyphosis and internal fixation loosening. Cobb angle was respectively(17.6+/-2.3) degrees, (18.1+/-2.7) degrees before treatment and last follow-up (P>0.05). MRI showed abscess was absorbed and spinal inflammation subsidised. VAS score was respectively 6.5+/-1.7, 1.4+/-0.5 before treatment and last follow-up (P<0.05). Seven patients had complications relating with drug adverse reaction,after discontinuation and treated with clinical symptom,the patients recovered normally. CONCLUSION: Anti-TB drugs alone can obtain satisfactory effects in treating early senile spinal tuberculosis, but strict supervision and individual administration should not be disregardful.

Outcomes of different surgical procedures in the treatment of spinal tuberculosis in adults.

OBJECTIVE: To investigate surgical methods and outcomes in the treatment of spinal tuberculosis (TB) in adults. SUBJECTS AND METHODS: One hundred and eighty-one patients (average age 39 years) without multiple-level noncontiguous spinal TB were followed up for 22-72 months. The patients were divided into four groups according to surgical procedure on the basis of the position and extension of the foci: group A (74 cases): anterior radical debridement and strut grafting with instrumentation; group B (83 cases): posterior instrumentation and bone grafting with anterior radical debridement and strut grafting in a single- or two-stage procedure; group C (10 cases): extrapleural anterolateral decompression and strut grafting with posterior instrumentation in thoracic or thoracolumbar spine, and group D (27 cases): single-stage transforaminal decompression and posterior instrumentation and fusion. RESULTS: There was a significant decrease (p < 0.05) in mean preoperative (81%) Oswestry's Disability Index. Except for 24 patients with lumbosacral TB who were only instrumented posteriorly, kyphosis degrees were corrected by a mean of 11.5° in the anterior instrumentation group and 12.6° in the posterior instrumentation group (p < 0.01). The correction loss was 6.8° in the anterior instrumentation group and 6.1° in the posterior instrumentation group at the last follow-up (p < 0.01). CONCLUSION: The four surgical procedures obtained good results for correction and maintenance of the correction, clearance of the foci, decompression of the spinal cord and pain relief in the treatment of spinal TB in adults, providing that the operative indication is accurately identified. However, the posterior approach was superior to anterior instrumentation for correcting deformity and maintaining the correction.

[Outcomes of anterior versus posterior instrumentation under different surgical procedures in the treatment of thoracolumbar spinal tuberculosis in adults].

OBJECTIVE: To compare the outcomes of anterior verus posterior instrumentation under different surgical procedures in the surgical management of thoracolumbar spinal tuberculosis (TB). METHODS: Between January 2004 and December 2009, 241 adult patients with thoracolumbar spinal TB underwent radical debridement and strut grafting plus anterior or posterior instrumentation in single-stage or two-stages. The mean age was 39 years (range: 16 - 67). The mean follow-up period for 189 patients was 37 months (range: 22 - 72). Among them, 157 cases underwent > 3 weeks of chemotherapeutic regimen of isoniazid, rifampin, pyrazinamide and ethambutol and the remaining 32 were operated for neurological impairment after 6-18 h with the same chemotherapeutic regimen. Except for 8 patients with skip lesions undergoing hybrid anteroposterior instrumentation, anterior instrumentation was utilized in 74 patients (Group A) and posterior instrumentation in 107 patients (Group B). RESULTS: In both groups, local symptoms of all patients were relieved significantly 1-3 weeks postoperatively. And 10/14 cases (71%) in Group A and 14/19 cases (74%) in Group B with neurological deficits had excellent or good clinical outcomes (P > 0.05). The levels of erythrocyte sedimentation rates (ESR) returned from 43.6 mm/h and 42.4 mm/h preoperatively to normal at 8-12 weeks postoperatively. Kyphosis degrees were corrected by a mean of 11.5° in Group A and 12.6° in Group B (P < 0.01). The correction loss was 6.8° in Group A and 6.1° in Group B at the last follow-up (P < 0.01). Fusion rates of the grafting bone were 92.5% and 91.8% respectively at the final follow-up (P > 0.05). Severe complications did not occur. CONCLUSION: Either anterior or posterior instrumentation can obtain good results in correction and maintenance of deformity, clearance of foci, decompression of spinal cord and pain relief in the treatment of thoracolumbar spinal TB as long as the surgical indications are properly selected. Posterior instrumentation may be superior to anterior instrumentation in the correction and maintenance of deformity.

Outcomes of anterior and posterior instrumentation under different surgical procedures for treating thoracic and lumbar spinal tuberculosis in adults.

PURPOSE: There are few articles in the literature comparing outcomes between anterior and posterior instrumentation in the management of thoracic and lumbar spinal tuberculosis (TB). METHODS: Between January 2004 and December 2009, 217 adult patients, average age 39 (range 16-67) years with thoracic and lumbar spinal TB were treated by anterior radical debridement and fusion plus instrumentation, anterior radical debridement with fusion and posterior fusion with instrumentation, posterolateral debridement and fusion plus posterior instrumentation or transpedicular debridement and posterior fusion with instrumentation in a single- or two-stage procedure. We followed up 165 patients for 22-72 (mean 37) months. Of these, 138 underwent more than three weeks chemotherapy with isoniazid, rifampin, pyrazinamide and ethambutol, and the remaining 27 underwent operation for neurological impairment within six to 18 hours of the same chemotherapy regimen. In no case did relapse occur. Apart from eight patients with skip lesions treated by hybrid anterior and posterior instrumentation, anterior instrumentation was used in 74 patients (group A) and 83 patients (group B) were fixed posteriorly. RESULTS: In both groups, local symptoms were relieved significantly one to three weeks postoperatively; ten of 14 patients (71%) in group A and 14 of 19 (74%) in group B with neurological deficit had excellent or good clinical results (P > 0.05). Erythrocyte sedimentation rates (ESR) returned from 43.6 mm/h and 42.7 mm/h, respectively, preoperatively to normal levels eight to 12 weeks postoperatively. Kyphosis degree was corrected by a mean of 11.5° in group A and 12.6° in group B, respectively (P < 0.01). Correction loss was 6.8° in group A and 6.1° in group B at the last follow-up (P < 0.01). Fusion rates of the grafting bone were 92.5% and 91.8%, respectively, at final follow-up (P > 0.05). Severe complications did not occur. CONCLUSION: These results suggest that both anterior and posterior instrumentation attain good results for correction of the deformity and maintaining correction, foci clearance, spinal-cord decompression and pain relief in the treatment of thoracic and lumbar spinal TB providing that the operative indication is accurately identified. However, the posterior approach may be superior to anterior instrumentation to correct deformity and maintain that correction.

[Construction of epithelia membrane protein 1 gene-deficient in human fetal nucleus pulposus cell model by lentivirus -mediated RNA interference].

OBJECTIVE: To construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration. METHODS: The lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods. RESULTS: The lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01). CONCLUSION: Lentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.

[Surgical treatment of lower lumbar tuberculosis with different operative procedures].

OBJECTIVE: To evaluate the surgical indications and clinical effectiveness of different operative procedures in the treatment of lower lumbar tuberculosis. METHODS: From June 2001 to Oct 2008, 72 patients with lower lumbar tuberculosis were treated by different operative procedures. Including 38 males and 34 females, with an average age of 38.5 years old ranging from 16 to 70 years. The average duration of symptom was 6.8 months (ranging from 4 months to 2 years). A single vertebrae was involved in 10 patients,two contiguous vertebrae in 50 cases and three vertebrae in 12 cases. The average preoperative lordotic angle was 13.1 degree (ranging from -5.0 degrees to 34.0 degrees). Three different operative procedures included: (1) posterior debridement and posterolateral fusion and posterior instrumentation in 28 patients; (2) anterior radical debridement and anterior fusion and anterior instrumentation in 32 patients; (3) image-guided percutaneous drainage (PCD) of tuberculous abscesses in 12 patients. The selection of the procedure was made according to the degree of the lesions. The resolution of inflammatory process, bony fusion, correction of sagittal angles and JOA scores were used for evaluating the result of the surgery and the complications were analyzed. RESULTS: All patients were followed up from 1.5 to 8.0 years (means 3.6 years). PCD was an effective treatment in 11 out of the 12 patients, one required surgical debridement and fusion. Among them, 57 (95%, 57/60) patients were treated by open operation showed successful bony fusion. The complications maily included common iliac vein injury in 3 patients, dural tear in 2 patients, they were all cured by intro-or postoperative treatment. The average immediate post-operative lordotic angle was 27.3 degree (35.0 degrees to 16.0 degrees), the average lordotic angle was 25.6 degree (33.0 degrees to 15.0 degrees) at final follow-up. Preoperatively and at final follow-up, JOA scores were respectively (15.2 +/- 3.4), (25.6 +/- 2.4) (P<0.01). CONCLUSION: Different operative procedures should be selected to treat lower lumbar tuberculosis according to the degree of lesions. Aggressive surgical treatment was found helpful in the resolution of inflammatory process and correcting the loss of lordosis, preventing progression of kyphosis.

[Application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease].

OBJECTIVE: To explore the value of application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease. METHODS: Clinical datas of 13 patients with multi-segment lumbar degenerative disease (8 males and 5 females,ranging in age from 51 to 72 year with an average of 65.0) were retrospectively analyzed between April 2008 and May 2009. The involved area included L3-S1 in 7 cases, L2-S1 in 3 cases, L3-L5 in 1 cases, L4-S1 in 2 cases. All patients underwent decompression, dynamic stabilization with Bioflex system, according to the severity of degenerative disc with/without interbody fusion. The clinical effects were evaluated by VAS, ODI. ROM and fusion segments were also observed. RESULTS: The mean follow up period was 19.5 months (from 12 to 26 months). The mean operative time was 183.4 min (from 90 to 240 min) and the mean volume of blood loss was 610.2 ml (from 400 to 1 220 ml). The mean VAS score was 7.8 +/- 1.3 preoperatively, 2.3 +/- 0.9 postoperatively and 2.1 +/- 0.8 at the last follow up. The average ODI was (60.50 +/- 4.40)% preoperatively, (17.80 +/- 2.10)% postoperatively and (16.20 + 2.40)% at the last follow up. The VAS and ODI significant improved in postoperatively (P < 0.05), and there was no statistical difference between postoperative and last follow up (P > 0.05). ROM of whole lumbar and non-fused segment showed obviously decreased and adjacent segment showed insignificant increased. The fusion rate of interbody fusion level was 95.0% (19/20). CONCLUSION: The preliminary clinical results show the Bioflex system combined with intebody fusion is a safe and effective technique in treating multi-segment lumbar degenerative disease.

[Surgical treatment for the thoracolumbar spinal tuberculosis with paraplegia].

OBJECTIVES: To explore the long-term effects,operating opportunity,indication of anterior debridement and decompression and bone graft in treating thoracolumbar tuberculosis with paraplegia. METHODS: From March 2006 to September 2008, the data of 56 patients with spinal tuberculosis were retrospectively analyzed. They were 36 cases male and 20 cases female, the age in ranging from 19 to 78 years with the mean of 42.5 years; course of disease in ranging from 3 months to 4 years. The lesion level was in T11-L2, including one stage in 6 cases, two stages in 42 cases, three stages in 7 cases, four stages in 1 case. According to the Frankel grade, grade A in 5 cases, B in 25, C in 21, D in 5. Preoperative Cobb angle had 30 cases under 30 degrees, 24 cases in 30 degrees-60 degrees, 2 cases more than 60 degrees [averaged in (35.0 +/- 3.3) degrees]. The mean VAS was 7.3 +/- 2.1 preoperatively. All patients had chemotherapy for 2-3 weeks. The clinical effects were evaluated by Cobb angle, VAS, condition of Frankel grade and fusion segments. RESULTS: All patients were followed up from 12 to 38 months with an average of 24 months. All wound obtained primary healing. The Cobb angle was(19.0 +/- 5.5) degrees immediately after operation, and lost (1.8 +/- 0.7) degrees at the follow up. The mean VAS was 1.4 +/- 0.3 postoperatively and 1.3 +/- 0.4 at final follow up. The Cobb angle and VAS significant improved in postoperatively (P < 0.05), and there was no statically difference between after operation and last follow up(P > 0.05). At 1 year after operation, all patients obstained successful bony fusion. Among 51 cases incomplete paraplegia recovered partially or completely (over Frankel D) through chemotherapy and rehabilitation training for 1-1.5 years; 5 cases of complete paraplegia had partially recovered, 3 cases from Frankel A to C, 2 cases from Frankel A to B. CONCLUSION: Base on the chemotherapeutic effect, paraplegia reason and location, total body condition to select different operative opportunity can effectively relieve spinal compression, stabilize spinal column, correct kyphosis and promote recovery of paraplegia.