Epidural fibrosis is the main cause of failed back surgery syndrome. To investigate the role of miR-146 in the diagnosis and development of epidural fibrosis. Lumbar disc tissues were collected from 72 lumbar disc herniation patients (45 developed epidural fibrosis and 27 did not). The expression of miR-146 in collected tissues and isolated epidural fibroblasts was detected by RT-qPCR. The relative levels of pro-inflammatory cytokines were analyzed by ELISA. The effect of miR-146 on the proliferation of fibroblasts was evaluated by MTT assay. miR-146 was significantly upregulated in epidural fibrosis patients compared with control patients. The expression of miR-146 was closely associated with the location, lower limb symptom and duration of disease of epidural fibrosis patients, and was positively correlated with the relative levels of pro-inflammatory cytokines. Moreover, miR-146 could discriminate epidural fibrosis patients from control patients. In isolated epidural fibroblasts, the overexpression of miR-146 dramatically enhanced its proliferation and the inflammatory response. miR-146 serves as a diagnostic biomarker for the early detection of epidural fibrosis. The upregulation of miR-146 enhanced the fibroblasts proliferation and inflammatory response in epidural fibrosis. This study provides a novel potential therapeutic target for epidural fibrosis.

IntroductionSkin scar is a common cutaneous complication, the outcome of which is unpleasant. Several microRNAs (miRs) participate in the process of skin scar formation. We aimed to explore the role of miR-27a-3p in NIH/3T3 mouse fibroblasts as well as the downstream protein and signaling cascades.Material and methodsmiR-27a-3p was aberrantly expressed in NIH/3T3 cells, followed by measurements of cell viability, migration and expressions of proteins related to proliferation and migration. Perlecan expression in cells aberrantly expressing miR-27a-3p was examined by Western blot analysis. Reporter gene assay was conducted to assess the relationship between miR-27a-3p and perlecan. Then, whether miR-27a-3p affected NIH/3T3 cells through regulating perlecan was ascertained. The effects of aberrantly expressed miR-27a-3p and perlecan on expression levels of VEGF, bFGF and key kinases in the MAPK/ERK and the PI3K/AKT pathways were detected.ResultsCell viability and migration were enhanced and protein expression levels of Cyclin D1, MMP-2 and MMP-9 were up-regulated by miR-27a-3p overexpression in NIH/3T3 cells. Then, we found that perlecan was positively correlated with miR-27a-3p expression, and its knockdown abrogated the effects of miR-27a-3p overexpression on NIH/3T3 cells. Finally, we found that the expression levels of VEGF and bFGF as well as phosphorylated levels of MAPK, ERK, PI3K and AKT were increased by miR-27a-3p overexpression, and those increases were reversed by perlecan knockdown.ConclusionsmiR-27a-3p promotes proliferation and migration of NIH/3T3 cells through up-regulating perlecan expression. Meanwhile, miR-27a-3p up-regulates expression levels of VEGF and bFGF, and activates MAPK/ERK and PI3K/AKT pathways through up-regulating perlecan expression.

Blumea balsamifera (L.) DC., a medicinal plant with high economic value in the Asteraceae family, is widely distributed in China and Southeast Asia. However, studies on the population structure or phylogenetic relationships with other related species are rare owing to the lack of genome information. In this study, through high-throughput sequencing, we found that the chloroplast genome of B. balsamifera was 151,170 bp in length, with a pair of inverted repeat regions (IRa and IRb) comprising 24,982 bp, a large single-copy (LSC) region comprising 82,740 bp, and a small single-copy (SSC) region comprising 18,466 bp. A total of 130 genes were identified in the chloroplast genome of B. balsamifera, including 85 protein-coding, 37 transfer RNA, and 8 ribosomal RNA genes; furthermore, sequence analysis identified 53 simple sequence repeats. Whole chloroplast genome comparison indicated that the inverted regions (IR) were more conserved than large single-copy and SSC regions. Phylogenetic analysis showed that B. balsamifera is closely related to Pluchea indica. Conclusively, the chloroplast genome of B. balsamifera was helpful for species identification and analysis of the genetic diversity and evolution in the genus Blumea and family Asteraceae.

目的 对比分析颈后路减压并侧块螺钉固定治疗多节段重度脊髓型颈椎病的疗效.方法 对2009年1月至2012年12月因多节段重度脊髓型颈椎病入院的病例共93例.按照入院顺序将患者划分为三组,分别为A组、B组和C组,A组患者27例,接受单开门椎管扩大成形术(颈椎后路);B组患者32例,接受全椎板切除减压侧块螺钉内固定术(颈椎后路);C组患者34例,接受单开门椎管扩大成形侧块螺钉内固定术(颈椎后路).对比患者手术前和手术后的ROM(颈椎活动度)、颈椎曲度、JOA评分以及并发症的情况.结果 术后3个月,三组患者的JOA改善未见统计学差异,P>0.05;末次随访时,B组和C组患者的评分改善情况由于A组,组间存在统计学差异,P<0.05.对比颈椎曲度,B组和C组间差异始终无统计学意义,P>0.05;A组颈椎曲度丢失率高于B、C两组(P<0.05).对比三组患者的颈椎活动度,组间数据无统计学差异,P>0.05.对比三组患者的轴性症状发生情况,B组和C组均低于A组(P<0.05).C5神经根麻痹三组差异无统计学意义(P>0.05).结论 在选择后路手术时,可优先考虑减压并侧块螺钉固定,并根据患者的实际情况和病情进行科学选择,保证患者的预后.

Osteosarcoma (OS) has a high incidence, malignity, and frequency of recurrence and metastasis. In this study, we aimed to explore the potential anti-cancer effects of Astragalus polysaccharides (APS) on human OS MG63 cells as well as underlying mechanisms. Viability of MG63 cells was assessed by CCK-8 assay to determine the adequate concentration of APS. Then, effects of APS on MG63 cell proliferation, cell cycle distribution, apoptosis, and migration and invasion were analyzed by BrdU incorporation, PI staining, flow cytometry, and transwell assays, respectively. The expression levels of proteins involved in these physiological processes were assessed by western blot analysis. Afterwards, miR-133a level in APS-treated cells was determined by qRT-PCR, and whether APS affected MG63 cells through regulation of miR-133a was determined. Finally, the activation of c-Jun N-terminal protein kinase (JNK) pathway was detected. We found that APS treatment suppressed the viability, proliferation, migration, and invasion of MG63 cells, as well as induced cell apoptosis. Moreover, APS enhanced the expression of miR-133a in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced MG63 cell proliferation, migration and invasion inhibition, as well as cell apoptosis. Furthermore, APS inactivated JNK pathway in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced inactivation of JNK pathway in MG63 cells. To conclude, APS repressed proliferation, migration, and invasion while induced apoptosis of OS MG63 cells by up-regulating miR-133a and then inactivating JNK pathway.

卵黄囊瘤是一种少见的起源于生殖细胞的生殖细胞瘤,常见于儿童及青少年,多发于卵巢、睾丸及骶尾部,主要沿身体中线分布,发生于其他部位少见,颅内及椎管内者极为罕见。我们收治1例颅内及椎管内髓外卵黄囊瘤患者,报道如下。患者女性,28岁。因"腰腿痛1月余,加重1周"于2016年11月28日收入我院。患者既往体健,曾于1个月前因腰痛伴右下肢疼痛,来我院就诊,初步检查后考虑"肌筋膜炎",给予对症治疗,效果较差,伴随双下肢进行性肌

PURPOSE:Acute paraplegia due to thoracic intervertebral disc protrusion and calcification is rare. The purpose of this study was to report two cases with acute paraplegia due to a calcified thoracic disc prolapse, and discuss its clinical diagnosis and surgical treatment with literature reviews.METHODS:These two cases were verified by patient history, physical examination, laboratory examination, CT and MRI studies, and pathological findings.RESULTS:CT scan revealed disc calcification and protrusion at the T11-12 level in case 1 and at the T10-11 level in case 2, respectively. MRI images revealed severe spinal cord compression with a hyperintense central core and surrounding hypointense area in two cases, which were directly connected to the calcified intervertebral nucleus pulposus. Pathological examination revealed calcium deposition. Patients underwent discectomy followed by interbody fusion, and satisfactory therapeutic outcomes were obtained.CONCLUSIONS:We suggest that decompression surgery should be carried out as early as possible for patients with early spinal myelopathy or paraplegia caused by a calcified protruded disc.

Objective To investigate the effects of Leptin on the base of anthropometry,densitometry,and biochemistry and reveal the role of Leptin in bone metabolism in postmenopausal women with osteoporosis (OP).Methods From January 2008 to January 2014,data of 82 postmenopausal females with or without OP were analyzed.They were randomized into two groups:OP (n=40) and conditional control group (CON) (n=42).The expression of Leptin receptors in osteoblasts was observed by immunohistochemistry.1 × 10 ng/ml,1 × 102 ng/ml,1 × 103 ng/ml,1× 104 ng/ml of Leptin were added respectively.Then the proliferation,differentiation and mineralization of osteoblasts were measure at 24 h,48 h and 96 h.The gene expressions of bone marrow mesenchymal stem cells (BMSCs) derived osteoblasts and adipocytes treated with Leptin were examined by quantitative analysis of Real time-PCR.Results The expression of Leptin receptors in osteoblasts was observed on plasma membranes and cytoplasm in the normal group and osteoporosis group.In osteoporosis group,different concentrations of Leptin enhanced cell proliferation,cell differentiation and cell mineralization.The growth rate of MTT and the concentration of ALP in the serum of were significantly increased with the effects of 1×10 ng/ml,1×102 ng/ml and 1×103 ng/ml of Leptin,and the action of 1×102 ng/ml Leptin was the most powerful with time dependence.Within 96 hours,the growth rate of osteoblasts increased gradually,and the concentration of ALP increased gradually in 3 weeks.The expression of RANKL/OPG in bone marrow BMSCs derived osteoblasts treated with Leptin were significantly increased.Conclusion Leptin receptor is present in osteoblasts,and Leptin affects biological behaviors of osteoblasts through receptors.The direct effect of Leptin may relate the expression of RANKL/OPG.Then,the balance between bone resorption and bone formation was broken and finally osteoporosis occurred.Lepin may promote osteogenesis and inhibit bone resorption in the differentiation of BMSCs.

BACKGROUND:Cervical Open-door laminoplasty with unilateral mass screw fixation and laminectomy with bilateral mass screw fixation are both procedures that treat cervical spondylotic myelopathy by expanding the space available for the spinal cord. Effectiveness and safety of two operative methods remain unclear. OBJECTIVE:To investigate the biocompatibility of implant and host between open-door laminoplasty with unilateral mass screw fixation and laminectomy with bilateral mass screw fixation to treat multi-segment cervical spondylotic myelopathy. METHODS: Data of 117 patients with multi-segment cervical spondylotic myelopathy (≥ 3 segments) were retrospectively analyzed. Sixty-five cases underwent open-door laminoplasty with unilateral mass screw fixation (laminoplasty group). Fifty-two cases underwent laminectomy with bilateral mass screw fixation (laminectomy group). Clinical outcomes were evaluated by the Japanese Orthopaedic Association scoring system and by recovery rate. Cervical curvature index and cervical range of motion were assessed by X-ray films in both groups. RESULTS AND CONCLUSION:The average folow-up time was 28 months (range 12-59 months) in both groups. No C5 nerve root palsy occurred in the two groups. Japanese Orthopaedic Association scores were greater during final folow-up than pre-fixation in both groups (P < 0.01). No significant difference in Japanese Orthopaedic Association score, recovery rate, and final folow-up cervical curvature index was detected in both groups (P > 0.05). Cervical range of motion was lower during final folow-up than pre-fixation in both groups (P < 0.01). Results confirmed that open-door laminoplasty with unilateral mass screw fixation and laminectomy with bilateral mass screw fixation have similar effectiveness in the improvement of neurological function, relieving pain and reducing complications, but wil greatly reduce the cervical range of motion.

目的 探讨人发角蛋白人工腱移植修复四肢肌腱、韧带缺损和周围神经损伤后功能重建的疗效. 方法 1998年4月至2010年3月对106例患者行人发角蛋白人工肌腱移植修复四肢肌腱、韧带缺损和周围神经损伤后功能重建术,男81例,女25例;年龄16 ～ 62岁,平均36岁.术后从切口愈合标准及分级、切口局部反应、患肢功能恢复时间分级、患肢肌力功能测定、关节功能恢复等方面评价疗效. 结果 术后所有患者随访2～11年.有8例患者切口出现皮下积液,换药7～42 d后愈合,其余98例切口均Ⅰ期愈合,愈合率达92.5％;切口局部均无疼痛、不适等感觉,无排斥反应;术后6～8周均可达到正常功能;94例患者肌力达3级以上,所有患者术后患肢肌力测定:优94例,良9例,可3例,优良率为97.2％;关节功能恢复测定:优92例,良10例,可4例,优良率为96.2％.结论 人发角蛋白人工腱材料对机体的刺激性小,多数患者切口可Ⅰ期愈合,无明显排斥反应,无粘连,术后患肢功能恢复快、效果好,是目前较为理想的肌腱移植替代材料.

目的 研究基底膜蛋白多糖对成骨细胞增殖及其分泌骨形态发生蛋白2(BMP-2)的影响.方法 体外培养大鼠成骨细胞,随机分为基底膜蛋白多糖抑制剂组(A组)、空白对照组(B组)、低氧状态下基底膜蛋白多糖抑制剂组(C组)、低氧状态下对照组(D组).各组分别进行相应处理,培养24、48、72、96 h.采用细胞增殖检测方法(CCK-8法)和ELISA法,检测并比较各组成骨细胞增殖和分泌BMP-2的变化.结果 培养24、48 h,A组与B组比较、C组与D组比较成骨细胞增殖活性均降低,差异有显著性(t=4.51～18.04,P＜0.05);培养72 h,各组差异无显著性(P＞0.05).培养96 h内,A组与B组比较、C组与D组比较成骨细胞分泌BMP-2降低,差异有显著性(t =6.61～30.50,P＜0.05).结论 在成骨细胞增殖早期基底膜蛋白多糖对成骨细胞增殖活性有显著影响,并对成骨细胞分泌BMP-2有促进作用.

Objective To evaluate the safety and feasibility of implanting occipital condyle screws in cadavers.Methods Screws of 3.5 mm in diameter and 20.0 mm in length were implanted into the ambilateral occipital condyles in 10 cadaveric specimens.CT scanning was performed after the procedure on the occipitocervical region for three-dimensional reconstruction of the region.In the three-dimensional occipitocervical models,relationships between the occipital condyle screw and its important surrounding tissues,like vertebral artery,spinal cord and hypoglossal canal,were measured.The length,upper and inside tilting angles of the screw,and distances between the screw and the hypoglossal canal and between the condylar entry point and the medial wall of the occipital condyle were also measured.Results Three-dimensional reconstruction models showed that all the occipital condyle screws were successfully implanted into the 20 occipital condyles in 10 cadaveric specimens.There was no injury to the spinal cord,nerve root,hypoglossal nerve or vertebral artery.At the left occipital bone,the average length,upper tilting angle and inside tilting angle of the condylar screws were respectively 20.96 ± 0.87 mm,31.00° ± 1.59° and 11.24° ± 0.71 °; the distances averaged 4.84 ±.0.52 mm between the condylar screw and the hypoglossal canal and 5.13 ±.0.79 mm between the condylar entry point and the medial wall of the occipital condyle.At the right occipital bone,the average length,upper tilting angle and inside tilting angle of the condylar screws were respectively 20.59±0.54 mm,30.85° ± 0.97° and 11.11 °±0.61°; the distances averaged 4.70±0.27 mm between the condylar screw and the hypoglossal canal and 5.04 ± 0.68 mm between the condylar entry point and the medial wall of the occipital condyle.Conclusions A surgeon who is familiar with the anatomy and skillful in operation can use occipital condyle screws in occipitocervical fusion without any injury to the important surrounding tissues.The screws,with definite parameters regarding entry point,course and length,can be used safely as an alternative or a modification of the conventional occipitocervical fusion in clinic.

Objective To investigate the effect of interference of perlecan expression on generation of type Ⅰ collagen in epidural scar cells.Methods Thirty SD male rats,weighing 250 grams and 2 months of age,were subjected to complete posterior lumbar laminectomy on L4-5.The epidural scar tissues were sampled at 20 days postoperatively for cellular culture by adherence.The third passage fibroblasts derived from the cellular culture were randomized into 3 groups (6 samples in each group):group A subjected to interference of perlecan expression by lentivirus mediated shRNA,group B subjected to interference of perlecan expression by lentivirus without shRNA mediation and group C,the blank control.Then hematoxylin-eosin staining (HE) was done to observe the formation of epidural scar and the activity of fibroblasts and the quantity of collagen synthesis.Western Blot and RT-PCR were performed to quantify the expression of perlecan and generation of type Ⅰ collagen after interference by lentivirus mediated shRNA.Comparisons were done between the 3 groups.Results The multiplicity of infection of 100 was the optimal when the cultured cells exhibited normal morphology and clear outline.RT-PCR showed the ratio of absorbance of the target bands of the type Ⅰ collagen RNA to that of the reference bands was 0.043 ± 0.003 in group A,0.473 ±0.011 in group B and 0.475 ± 0.013 in group C.Western Blot showed the ratio of absorbance of the target bands of the type Ⅰ collagen RNA to that of the reference bands was 0.190±0.005 in group A,2.116±0.012 in group B and 2.122±0.009 in group C.There were significant differences between group A and groups B and C regarding the above indexes (P ＜ 0.05).The generation of type Ⅰ collagen in group A was significantly lower than in groups B and C (P ＜ 0.05).Conclusion Since the interference of perlecan expression by lentivirus mediated shRNA can reduce the expression of type Ⅰ collagen in epidural scar fibroblasts,it can be used to inhibit epidural scar formation after laminectomy.

Objective To observe the effect of pelercan on proliferation of fibroblasts and its relationship with basic fibroblast growth factor (bFGF).Methods The rat model was established,and the scar connective tissue was collected and subjected to the primary culture of rat fibroblasts.Lentivirus was used to downregulate the expression of pelercan.Fluorescent quantitative polymerase chain reaction (FQ-PCR),Western blotting and methyl thiazol tetrazolium (MTT) assay were performed to detect the expression of perlecan and cell proliferation rate.Results After the lentivirus transfection,the perlecan mRNA expre ssion level in fibroblasts was reduced by (75.1 ± 1.1) ％ in perlecan-interfering group.In low-serum condition,the cell proliferation rate was decreased by (40.6 ± 3.1)％ in perlecan-interfering group.Treatment with 1 μg/L bFGF had no significant effect on the cell proliferation (P ＞ 0.05).Conclusion Infection of rat fibroblasts with lentivirus down-regulated the expression of perlecan and bFGF.Cell proliferation rate and sensitivity of rat fibroblasts to bFGF were both reduced when perlecan was downregulated.

Objective: Epidural scar,also known as epidural fibrosis,means scar tissue or fibrosis formed in the surgery within the scope of the epidural,and it is body's trauma repair response.Scar adhesion and contraction will stretch the dura mater and nerve root,limit their activities,scar encysted nerve root may be abnormal stretched and squeezed,nerve fiber's axoplasmic transportation,arterial blood supply and venous return suffocated,nerve root and dorsal ganglion is very sensitive to mechanical pressure.Therefore,it will produce a series of symptoms,such as pain,numbness,lower extremity muscle strength is reduced.Recently,most of the research on the prevention of epidural scar is how to reduce the occurrence of postoperative scar adhesions(which leads to the occurrence of complications) through physical or chemical barrier after the laminectomy.But,the research on inhibit scar's main constituent in the process of scar information to reduce epidural scar formation after laminectomy is few.Inhibition epidural scar formation by reducing typeⅠ collagen production.Methods: 30 SD rats were randomly and evenly divided into six groups and full laminectomy was performed on the 4th,5th lumbar vertebra in each experiment.The epidural scar were sampled at the 1st,2nd,3rd,4th,5th and 6th weeks postoperatively,one group each week.Then hematoxylin-eosin staining(HE) was done to observe the formation of epidural scar and the activity of desmocyte and the quantity of collagen synthesis.Cell culture was done to observe the desmocyte morphology and activity.Western Blot and RT-PCR were made to quantify the expression of the perlecan and type I collagen after interference by lentivirus mediated shRNA interference.Results: According to hematoxylin-eosin staining(HE) and cell culture,scar desmocytes at the second and third weeks are the most suitable for the experiment.Western Blot and RT-PCR showed that type I collagen expression reduced compared to control group after the perlecan was interference by lentivirus mediated shRNA interference(RT-PCR F=509.331;q A,B=-43.371,P<0.01;q A,C=-46.133,P<0.01;Western-Blot F=337.578;q A,B=-112.433,P<0.01;q A,C=-89.227,P<0.01).Conclusion: Type I collagen expression reduced after the perlecan was interfered by lentivirus mediated shRNA,epidural scar can be inhibition by this way.Compared with the control group,through shRNA(which was mediated by lentivirus)interference perlecan(in fibroblast),the formation of collagen typeⅠhas statistically significant difference.Indicate that by inhibiting the expression of perlecan in hypertrophic scar fibroblast can effectively reduce the generation of type I collagen protein.Not only because type I collagen is the major constituents of scar,but also because the increase of type I collagen content leads to scarless healing converted to scar healing.This method is theoretically able to inhibit effectively andreduce epidural scar formation.Therefore,the theory of inhibiting formation of epidural scar by interfering perlecan expression is feasible,it also laid a theoretical foundation for further in vivo test to inhibition the formation of epidural scar after laminectomy.

Objective To research the proliferation of fibroblasts in epidural scar transfected by perlecan shRNA lentiviral particles.Methods Experimental models were established in 30 Wistar mice which underwent laminectomy.The epidural scar tissue was harvested after culture for 4 days.The fibroblasts in the epidural scar tissue were cultured by tissue explant adherence before they were randomly assigned into no transfection (control group),green fluorescent protein (GFP) lentiviral transfection (GFP group) and shRNA lentiviral transfection (shRNA group).After the fibroblasts were successfully transfected,their expressions of perlecan mRNA and protein and proliferation were detected by Western-blot,RT-PCR and MTT and compared as well among the 3 groups.Results Western-blot and RT-PCR showed the expressions of perlecan mRNA and protein were significantly lower in the shRNA group than in the control and GFP groups (P ＜ 0.05).MTT showed no significant difference among the 3 groups in the optical density at 0 hour (P ＞0.05) but significantly lower optical density in the shRNA group than in the control and GFP groups at 24 to 96 hours (P ＜ 0.05).Conclusion Growth of fibroblasts in epidural scar can be inhibited significantly by transfection with perlecan shRNA lentiviral particles.

BACKGROUND:Many types of steel plates are used for internal fixation of calcaneal fractures through extensive lateral approach. The fixation screw at the anterior calcaneal process must be placed into the dense compression trabeculae located directly under the calcaneocuboid articular surface to achieve a stable fixation. METHODS:The transverse diameter and inner tilt angle of the calcaneocuboid articular surface were measured and the inner structures near the calcaneocuboid articular surface were observed in forty adult calcaneus bone specimens to provide an anatomical basis for internal fixation of calcaneal fractures. RESULTS:The transverse diameter was 22.67 ± 2.14 mm and the inner tilt angle was 60.4 ± 7.1°. CONCLUSION:Screws should be implanted under the calcaneocuboid articular surface and the length and direction of the screw should be selected according to the transverse diameter of the calcaneal articular surface and the inner tilt angle, respectively.

目的比较手术、保守治疗颈椎间盘突出症的效果。方法 605例颈椎间盘突出症患者均合并头晕。其中302例(手术组)采用颈椎前路髓核摘除减压自体髂骨植骨加自锁钢板内固定治疗(手术组),303例(保守组)采用保守疗法治疗。比较两组疗效。结果随访2个月～2年,保守组治愈86例、显效80例、有效61例、无效76例,治愈率28.3%,总有效率74.9%;手术组治愈278例、显效例13、有效6例、无效5例,治愈率92.0%,总有效率98.3%。手术组治愈率和总有效率均明显高于保守组,P均<0.05。结论颈椎前路髓核摘除减压自体髂骨植骨加自锁钢板内固定术治疗颈椎间盘突出症的疗效优于保守疗法。

Objective To make a research on the expression of perlecan in the formation of epidural scar after posterior lumbar laminectomy in the rat.Methods Sixty Wistar rats were randomly and evenly divided into five groups:the experimental groups including A(the first week after surgery),B(the second week),C(the fourth week),D(the sixth week)and the control group.Laminectomy was performed on lumbar vertebra in each experimental group.The hematoxylin-eosin(HE)staining was used to observe the formation of epidural scar.Immunohistochemistry was performed to detect the perlecan located in the epidural scar.Western blot and RT-PCR were made to quantify the expression of the perlecan after surgery.Results According to HE,with prolonged,the scar tissue gradually formed.Immunohistochemistry demonstrated that the perlecan was expressed in the fibroblast.Western blot and RT-PCR showed the high level expression of perlecan in the experimental groups compared to the control group(P＜0.01).Furthermore,the expression of perlecan gradually increased with the prolongation of exposure time and had significantly statistical significance (P＜0.01)between the first three experimental groups.However,compared to the fourth week group,the expression of perlecan in the sixth week decreased obviously(P＜0.01).Conclusion Perlecan was expressed in the fibroblast of the epidural scar,and which plays an important role in the formation of epidural scar.

Objective To investigate the impact of stable expression of perlecan shRNA lentiviral particles on proliferation of NIH3T3 cells. Methods Mouse fibroblasts were cultured.Lentiviral particles-green fluorescent protein (LV-GFP) was used to transfect the cultured NIH3T3 cells with multiplicity of infection (MOI) of 10,30 and 50.The GFP expression was observed with fluorescence microscopy after transfection for one week to estimate the proper MOI and the time of GFP expression needed.The transfection efficiency of LV-GFP with the proper MOI by fluorescence-activated cell sorting was detected.The stably transfected cell lines were developed by puromycin screening for more than 2 weeks.The third generation HFF in good condition was randomly divided into 3 groups:GFP group,shRNA group and control group.RT-PCR,Western blot and MTT assays were used to detect the expressions of perlecan mRNA and protein and cell proliferation in the 3 groups. Results Perlecan mRNA and protein showed high expressions in the control and GFP groups but low expressions in the shRNA group,with significant differences respectively between the shRNA group and the other 2 groups ( P ＜ 0.05).There was no significant difference between the 3 groups in the optical density at the first 2 days ( P ＞ 0.05).On 3 to 6 days the cells in the control and GFP groups grew normally while the cells in the shRNA group proliferated in a weak manner.the transfected cells in the shRNA group showed a significantly reduced proliferation rate compared with the other 2 groups ( P ＜ 0.05 ). Conclusion The growth of NIH3T3 cells can be inhibited significantly by transfection with perlecan shRNA lentiviral particles.