Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults (version 2023) / 中华创伤杂志

The acute combination fractures of the atlas and axis in adults have a higher rate of neurological injury and early death compared with atlas or axial fractures alone. Currently, the diagnosis and treatment choices of acute combination fractures of the atlas and axis in adults are controversial because of the lack of standards for implementation. Non-operative treatments have a high incidence of bone nonunion and complications, while surgeries may easily lead to the injury of the vertebral artery, spinal cord and nerve root. At present, there are no evidence-based Chinese guidelines for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults. To provide orthopedic surgeons with the most up-to-date and effective information in treating acute combination fractures of the atlas and axis in adults, the Spinal Trauma Group of Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field of spinal trauma to develop the Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults ( version 2023) by referring to the "Management of acute combination fractures of the atlas and axis in adults" published by American Association of Neurological Surgeons (AANS)/Congress of Neurological Surgeons (CNS) in 2013 and the relevant Chinese and English literatures. Ten recommendations were made concerning the radiological diagnosis, stability judgment, treatment rules, treatment options and complications based on medical evidence, aiming to provide a reference for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults.

Clinical guideline for diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture (version 2023) / 中华创伤杂志

Ankylosing spondylitis (AS) combined with spinal fractures with thoracic and lumbar fracture as the most common type shows characteristics of unstable fracture, high incidence of nerve injury, high mortality and high disability rate. The diagnosis may be missed because it is mostly caused by low-energy injury, when spinal rigidity and osteoporosis have a great impact on the accuracy of imaging examination. At the same time, the treatment choices are controversial, with no relevant specifications. Non-operative treatments can easily lead to bone nonunion, pseudoarthrosis and delayed nerve injury, while surgeries may be failed due to internal fixation failure. At present, there are no evidence-based guidelines for the diagnosis and treatment of AS combined with thoracic and lumbar fracture. In this context, the Spinal Trauma Academic Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate the Clinical guideline for the diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture ( version 2023) by following the principles of evidence-based medicine and systematically review related literatures. Ten recommendations on the diagnosis, imaging evaluation, classification and treatment of AS combined with thoracic and lumbar fracture were put forward, aiming to standardize the clinical diagnosis and treatment of such disorder.

IL6 Receptor Facilitates Adipogenesis Differentiation of Human Mesenchymal Stem Cells through Activating P38 Pathway

Background and Objectives@#Mesenchymal stem cells (MSCs) have the multipotent capacity to differentiate into multiple tissue lineages as well as to self-renew, which is the main origin of adipocytes. IL6/IL6R pathway exerts a significant role in tissue regeneration and cell differentiation. Whereas, the underlying mechanism between IL6/IL6R pathway and MSCs adipogenesis differentiation remains elusive. @*Methods@#MSCs from healthy donors were cultured in adipogenesis differentiation medium for 0∼14 days, during which their adipogenesis differentiation degree was evaluated by Oil Red O staining. The expression of IL6R was detected in MSCs during adipogenesis differentiation. Knockdown and overexpression of IL6R were respectively performed using siRNA and lentivirus to investigate its effect on MSCs adipogenesis differentiation. The adipogenesis marker genes expression and MAPK pathway activation were detected by Western blotting. The role of P38 pathway in the adipogenesis differentiation of MSCs was determined using the specific inhibitor SB203580. @*Results@#The expression of IL6 and IL6R increased during adipogenesis differentiation in MSCs, which were positively correlated with Oil Red O quantification result. Knockdown and overexpression experiments demonstrated a positive correlation between the expressions of IL6R and MSCs adipogenesis differentiation, accompanied by same trend of P38 phosphorylation. Besides, the specific P38 inhibitor SB203580 markedly inhibited the adipogenesis differentiation potential of MSCs. @*Conclusions@#This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.

Clinical guideline for surgical treatment of symptomatic chronic osteoporotic vertebral fractures / 中华创伤杂志

According to the pathological characteristics of symptomatic chronic thoracic and lumbar osteoporotic vertebral fracture (SCOVF), the different clinical treatment methods are selected, including vertebral augmentation, anterior-posterior fixation and fusion, posterior decompression fixation and fusion, and posterior correction osteotomy. However, there is still a lack of a unified understanding on how to choose appropriate treatment method for SCOVF. In order to reflect the new treatment concept and the evidence-based medicine progress of SCOVF in a timely manner and standardize its treatment, the clinical guideline for surgical treatment of SCOVF is formulated in compliance with the principle of scientificity, practicability and advancement and based on the level of evidence-based medicine.

Elevated TRAF4 expression impaired LPS-induced autophagy in mesenchymal stem cells from ankylosing spondylitis patients

Ankylosing spondylitis (AS) is a type of autoimmune disease that predominantly affects the spine and sacroiliac joints. However, the pathogenesis of AS remains unclear. Some evidence indicates that infection with bacteria, especially Gram-negative bacteria, may have an important role in the onset and progression of AS. Recently, many studies have demonstrated that mesenchymal stem cells (MSCs) dysfunction may contribute to the pathogenesis of many rheumatic diseases. We previously demonstrated that MSCs from AS patients exhibited markedly enhanced osteogenic differentiation capacity in vitro under non-inflammatory conditions. However, the properties of MSCs from AS patients in an inflammatory environment have never been explored. Lipopolysaccharide (LPS), a proinflammatory substance derived from the outer membrane of Gram-negative bacteria, can alter the status and function of MSCs. However, whether MSCs from AS patients exhibit abnormal responses to LPS stimulation has not been reported. Autophagy is a lysosome-mediated catabolic process that participates in many physiological and pathological processes. The link between autophagy and AS remains largely unknown. The level of autophagy in ASMSCs after LPS stimulation remains to be addressed. In this study, we demonstrated that although the basal level of autophagy did not differ between MSCs from healthy donors (HDMSCs) and ASMSCs, LPS-induced autophagy was weaker in ASMSCs than in HDMSCs. Specifically, increased TRAF4 expression in ASMSCs impaired LPS-induced autophagy, potentially by inhibiting the phosphorylation of Beclin-1. These data may provide further insight into ASMSC dysfunction and the precise mechanism underlying the pathogenesis of AS.

The perioperative risk factors of postoperative complications after posterior lumbar fusion operation: a retrospective analysis of 654 cases / 中华骨科杂志

Objective To analyze the perioperative risk factors of postoperative complications after posterior lumbar fusion operation.Methods The clinical data of 654 patients with posterior lumbar fusion during 2010 and 2014 were retrospectively analyzed.Using x2 test and one-way ANOVA,the predicted risk factors were screened for further Logistic regression.Results The total complication rate was 11.6％ among all 654 patients.The major complications included cardiac infarction,deep infection,sepsis,neurological impairment,and secondary operation.And the minor complications included wound dehiscence,urinary tract infection,pulmonary infection,gastrointestinal bleeding,CSF leakage and others.According to x2 test and one-way ANOVA,renal function insufficiency,preoperative neurological injury,ASA higher than Ⅲ level,intraoperative blood loss,long operation length,and usage of autogenous bone were screened as risk factors of complications.Renal function insufficiency,preoperative neurological injury,intraoperative blood loss,and long operation length were screened as risk factors of minor complications.And male,renal function insufficiency,preoperative neurological injury,intervertebral fusion,and posteriolateral fusion were screened as risk factors of major complications.However,according to Logistic regression,the independent risk factor of complications were preoperative neurological injury and long operation length;independent risk factors of minor complications were renal function insufficiency,preoperative neurological injury and long operation length;and independent risk factor of major complications was preoperative neurological injury.Conclusion Preoperative neurological injury,renal function insufficiency and long operation length are proved to be the risk factors of postoperative complication in lumbar fusion surgery.

Adjacent segment disease and degeneration after lumbar spine surgery / 中华骨科杂志

Lumbar spine fusion surgeries have adverse effects on adjacent segments and lead to adjacent segment disease and degeneration which increase the possibility of revision surgery and affect the final outcome.If new degeneration or aggravation of the primary degeneration in the adjacent segment only occurs in the images but without corresponding clinical symptoms,it would be called adjacent segment degeneration;if the corresponding clinical symptoms co-exist with degeneration in images,it would be called adjacent segment disease.Generally,the stress concentration on adjacent intervertebral discs and facet joints caused by spine fusion is the main pathogeny of adjacent segment degeneration and disease.The damage of normal spinal anatomy structure in surgery and the natural spine degeneration process are also important pathogenic factors.The occurrence of adjacent segment degeneration and disease after lumbar surgery is related to age,body weight,postoperative spinal-pelvic sagittal balance and the preoperative degeneration of adjacent segment.Surgical programs including numbers of the fusion segments,surgical approach and whether to adopt non-fusion technology have great effect on the occurrence of adjacent segment degeneration and disease.In the planning of surgeries,necessary measures and methods should be taken to prevent adjacent segment disease and degeneration according to the different patients.Non-fusion technology,minimally invasive spine surgery technique and Topping-off technique can help reduce the occurrence of adjacent segment disease and degenerative.If the patients are combined with high risk factors of adjacent segment degeneration and disease,more attention should be paid and appropriate and individualized therapies should be chosen.

Bone marrow mesenchymal stem cells derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages / 中国组织工程研究

BACKGROUND:Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is stil unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE: To explore the immunoregulation capability of bone marrow mesenchymal stem cels from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cels from healthy donors on ankylosing spondylitis. METHODS: Bone marrow mesenchymal stem cels were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cels were used in subsequent experiments. A human monocytic cel line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cels and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6 (TSG-6) proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the mRNA level of cytokines secreted by bone marrow mesenchymal stem cels and macrophages. RESULTS AND CONCLUSION:The typical mesenchymal stem cel surface markers were expressed in both bone marrow mesenchymal stem cels from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and mRNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cels from patients with ankylosing spondylitis were obviously higher than those from healthy donors (P < 0.05). TSG-6 secreted by bone marrow mesenchymal stem cels from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cels from healthy donors in both RNA transcriptional and protein levels (P < 0.05). Our study demonstrates that bone marrow mesenchymal stem cels from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting the tumor necrosis factor-α secretion from macrophages, which reveals a mechanism of immune disorder in ankylosing spondylitis. The therapeutic mechanism of bone marrow mesenchymal stem cels from healthy donors may work by secreting enough TSG-6 to inhibit the activation of macrophages in patients with ankylosing spondylitis, and thereby to decrease the secretion of tumor necrosis factor-α. Cite this article:Sun SH, Wang P, Su CY, Xie ZY, Li YX, Li D, Wang S, Su HJ, Wu XH, Deng W, Wu YF, Shen HY. Bone marrow mesenchymal stem cels derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):13-19.

Total en-bloc spondylectomy for recurrence spinal tumor / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical outcomes of total en-bloc spondylectomy (TES) in recurrence spinal tumor.</p><p><b>METHODS</b>The study was a retrospective study of recurrence spinal tumor from January 2010 to October 2013. A total of 6 patients with recurrent spinal tumor underwent TES procedures, with 5 cases located in thoracic spine and 1 case located in L1. There were 3 male and 3 female patients, with a mean age of 33.2 years. Pathological diagnosis included giant cell tumor of bone in 3 cases, breast cancer, lung cancer and nasopharyngeal carcinoma with 1 case in each. The operation time, bleeding loss, resected segments, cutting edge, spinal cord function and complications was evaluated.</p><p><b>RESULTS</b>Single segment resected in 1 case, 2 segments resected in 2 cases and 3 segments resected in 3 cases. The average operation time was 8.9 hours (7.5 to 12.0 hours). The average blood loss was 3 116 ml (2 500 to 4 500 ml). The average follow-up period was 23.2 months (12 to 47 months) without recurrence. There was no spinal cord injury during operation. The neurologic function was significantly improved in 2 cases (American Spinal Injury Association (ASIA) grade C to grade D), unchanged in 1 cases (ASIA grade B) and no deteriorated case in 3 cases (ASIA grade E). There was no perioperative deaths case. Complications included 2 cases pleural rupture, 1 case dural tear and 1 case massive haemothorax. No peri-operation death case.</p><p><b>CONCLUSION</b>Some of the recurrent spinal tumors are still suitable for en-bloc resection and TES procedure with the extent of its applicability under strict control.</p>

The perioperative risk of deep vein thrombosis and individualized anti-coagulation treatment in patients with hepatic cirrhosis undergoing total hip arthroplasty: a pilot study / 中华骨科杂志

Objective To investigate the perioperative risk of deep vein thrombosis (DVT) in patients with hepatic cirrhosis that underwent total hip arthroplasty (THA), and to evaluate the safety and feasibility of individualized anti-coagulation treatment.Methods There were 25 patients complicating hepatic cirrhosis that underwent THA (from Jan.to Dec.2014), including 17 males and 8 females, aged 57.9t9.2 years.The primary causes of THA were avascular necrosis of the femoral head (eighteen cases) and osteoarthritis of the hip (seven cases).Low molecular weight heparin (LMWH) was applied for anti-coagulation treatment.Parameters of hepatic function and coagulation function of THA cases (randomized thirty cases, from Jan.2008 to Dec.2008) without hepatic cirrhosis were used as reference for monitoring.For the cases of massive blood loss or upper gastrointestinal hemorrhage, a LMWH administration pause and an administration of fresh frozen plasma and clotting factors were performed in order to maintain a hemorrage/coagulation balance.The clinical outcome of the hip joint was evaluated and complications were treated.A subsequent follow-up was also carried out after perioperative period.Results All cases received successful surgeries and followed up.The follow-up duration was 34± 15.7 months.The preoperative Harris hip score was 32.4± 10.2 points, while the most recent follow-up score was 82.9±6.1 points, which was statistically significant.Dislocation, periprosthetic fracture and periprosthetic infection were absent.All cases received individualized anti-coagulation treatments during peripoerative period.A hemorrage/coagulation balance was achieved.The dynamic parameter curves did not present excessive deviation from reference.One case encountered intermuscular hematoma of the lower limbs 48 hours postoperatively, which was solved by a LMWH pause and administration of fresh frozen plasma and clotting factors.One case suffered upper gastrointestinal hemorrhage five days postoperatively, which was controlled by a LMWH pause and the administration of somatostatin and proton pump inhibitor.Jaundic got worse in one case three days postoperatively but got relieved after treatment.Overt blood loss was 686t141.8 ml.Perioperative death, hepatic failure, hepatic encephalopath, hepatorenal syndrome were absent.No DVT was observed.Conclusion There are risks of DVT in patients of hepatic cirrhosis.Individualized anti-coagulation treatment is needed during perioperative period of THA.

Allogeneic blood transfusion alleviates hip joint pain induced by ankylosing spondylitis / 中国组织工程研究

BACKGROUND:Pain is the main clinical manifestation for ankylosing spondylitis. At present, nonsteroid anti-inflammatory drugs are oral y taken, but the effects are limited and toxic and side effects are more. Thus, there is no effective scheme for treatment of pain induced by ankylosing spondylitis. OBJECTIVE:To investigate the correlation between postoperative joint pain al eviation and al ogeneic blood transfusion, and the mechanisms. METHODS:We retrospectively analyzed clinical data of 88 ankylosing spondylitis patients combined with kyphosis who received only one section of osteotomy surgery merging hip joint pain. We compared the visual analog scale score of hip joint and detected the variation of leucocytes, lymphocytes and immunoglobulin concentrations before and after the operation in the groups of fresh al ogeneic whole blood transfusion, autologous whole blood transfusion, and mixed transfusion of al ogeneic and autologous whole blood. Flow cytometry was used to analyze the number and ratio of peripheral blood Th17 cells and Treg cells which were both highly associated with autoimmune diseases. RESULTS AND CONCLUSION:The symptom of hip arthralgia obviously improved in both groups transfused by fresh al ogeneic whole blood or al ogeneic-autologous mixed whole blood. However, no obvious variation was detected in leucocytes, lymphocytes and immunoglobin concentration. However, flow cytometry results indicated that Th17/Treg proportion associated with autoimmune diseases was increased remarkably in peripheral blood of ankylosing spondylitis patients. Results suggested that al ogeneic whole blood transfusion can al eviate patients’ joint pain by correcting the imbalance of Th17/Treg which may improve their immune state.

Modified surgical techniques in total en bloc spondylectomy for thoracic and lumbar tumors with a single posterior approach / 中华骨科杂志

Objective To investigate the operation key points, instrument improvement and shortterm effects in total en bloc spondylectomy (TES) via a single posterior approach for thoracic and lumbar tumors. Methods A series of modified instruments have been designed for the TES, including threadwire saw (T-saw) with a diameter of 0.81 mm, director and clamping for the saw, L shape and furcation osteotomes.The corpectomy of original TES which was defined as "one step dissection" from anteriorly to posteriorly, was modified into "two step dissection" which means that corpectomy was performed with saw cutting anteriorposteriorly and the L shape cutting posterior-anteriorly. In the cases with difficulty in pediculotomy using a T-saw, furcation osteotome was used for pediculotomy. Ten patients with thoracic or lumbar tumors were treated with the modified TES. There were 1 case of bone giant cell tumor, 1 case of bone neurilemmoma and 8 cases of metastatic tumors. All patients suffered moderate-severe pain and neurological deficit. Results The average follow-up period was 8.1(3.3-18.1) months. The average operating time was 7.8 h(6.0-10.3 h),and average blood loss was 2100 ml (1200-3500 ml). No disruption of dural mater, the leakage of cerebrospinal fluid, iatrogenic spinal cord injury and major vessel damage occurred. Two patients who underwent pleura disruption happened during the operation were treated with intrathoracic drain remedy. Among 7 cases with thoracic tumors, significant improvement in neurological function were achieved in 5 patients with the improvement of one grade in ASIA classification, while no change was found in 2 cases. In 3 cases with lumbar tumor, lumbar nerve root pain relieved and the muscle strength had recovered to grade 4 at least postoperatively. Conclusion Significant improvement has been achieved in the maneuverability and safety of the modified surgical techniques in TES with a single posterior approach for thoracic and lumbar tumors.

Outcome and prognosis of avascular necrosis after talus fracture / 中华创伤杂志

Objective To study the incidence and prognosis of avascular necrosis after talus fracture. Methods A retrospective survey was performed in 12 patients ( 13 feet) with talus fractures admitted into hospital from July 2004 to November 2009 to analyze necrosis rate, ankle function recovery and disability rate. According to Hawkin' s classification system, there were two patients with type Ⅰ feet, four with type Ⅱ feet, five with type Ⅲ feet and two with type Ⅳ feet. Results All patients were followed up for average period of 19.6 months (range 11-52 months). Avascular necrosis was detected in eight feet, of which one foot was treated with ankle fusion, one with subtalar arthrodesis and one with bone implantation. The other five feet had good ankle and subtalar function, with no collapse or osteoarthritis. According to Maryland foot score, the result was excellent in eight patients, good in two, fair in one and failure in two, with excellence rate of 77%. Conclusion The incidence of avascular necrosis after talus fracture is related to the location and energy of trauma. However, the function prognosis of the talus shows no correlation with necrosis.

Osteogenic Activity of MSC Infected by Recombinant Adenovirus Vector Ad-LMP-1 / 中山大学学报(医学科学版)

[Objective]This study was designed to construct a recombinant adenovirus vector contains LMP-1 gene,and investigate the osteoinductive activity of MSC which were transfected recombinated adenoviral vector carrying LMP-1 gene.[Methods]Total RNA was extracted from mt osteoblast and the LMP-1 gene was acquired by RT-PCR,the LMP-1 gene and entry vector pENTR/D-TOPO were used to create the entry clone with the directional TOPO clone technology,then the entry clone and the expression vector were used to create the expression clone throush the LR recombination reaction.The adenovirus expression clone was linearized by PacI and transfected to the 293A cell line to harvest a high titer.Ad-LMP-1 was infected into the 3rd passage MSC,the expression of LMP-1 was detected by Western blot.The osteogenic activity of MSC was evaluated by the expression of collagen Ⅰ,ALP,osteocalcin and the formation of bone nodule.[Result]The LMP-1 gene was successfully acquired and confirmed,the entry clone and the expression clone were both verified by enzymes digestion,and the expression clone was further confirmed by sequenced.The expression of LMP-1 was detected successfully in MSC.The increasing expression of collagen Ⅰ,osteocalcin.ALP and bone nodule were observed by comparing to the control group.[Conclusion]Gateway technology not only make construction of the pAd-LMP-1 recombination adenovirus vector simple and fast,but also get a high transfection efficacy in MSC.LMP-1 gene can induce the osteoblast differention of MSCs,and improve its osteogenic activity.The adenovirus vector is reliable to be used in further gene therapy research.

Delayed transplantation of olfactory ensheathing cells for thoracic cord injury in adult rats / 中国组织工程研究

BACKGROUND: Spinal cord can regenerate after injury in certain microenvironment. Olfactory ensheathing cells (OECs)have the characteristics of astrocytes and Schwann cells and can accelerate the spinal cord axonal regeneration.OBJECTIVE: To make injured thoracic cord rat models and observe the effect of OECs on injured spinal cord axonal regeneration.DESIGN: Observational experiment.SETTING: Second Affiliated Hospital of Sun Yat-Sen University.MATERIALS: The experiment was performed at the Second Affiliated Hospital of Sun Yat-Sen University from January 2001 to November 2002.Totally 20 adult SD male rats with the body mass of (380±20) g were provided by Experimental Animal Center of Sun Yat-Sen University (number of institution license SYXK2004-0020). There were DMEM culture solution with low glucose (L-DMEM, GibcoBRL), fetal calf serum (FCS) (Hyclone), myelin basic protein (MBP) (Sigma) and nerve growth factor receptor antibody (Sigma). They were divided into cell transplantation group and control group by the method of random digits table with 10 in each group.METHODS: The adult SD rats were anaesthetized and decapitated to obtain the whole olfactory bulb and then isolate olfactory nerve with a sterile operation. Thoracic cord injury models were established by modified Allen method. 10μL OECs suspension (2.5×1010 L-1) was injected into injured spinal cord of the cell transplantation group, whereas DMEM/F12 (1:1) culture solution of the same dose was injected in the control group. The influence of OECs on spinal cord axonal regeneration was observed by histological and immunohistochemical method 6 weeks after transplantation.MAIN OUTCOME MEASURES: ①OECs were identified by nerve growth factor receptor antibody staining. ②Repair of myelin sheath was observed by MBP staining. ③Nerve axonal regeneration was observed by argentaffin staining. RESULTS: Two rats in the cell transplantation group and 3 rats in the control group died, so totally 15 rats were involved in the result analysis. ①In the cell transplantation group,injured spinal meninges were integrated,but spinal cord became thin as compared with the normal spinal cord. By hematoxylin-eosin (HE) staining, multipolar cells appeared in injured region and the cells were fused excessively with myeloid tissues. It proved that survival OECs were integrated well within the host. New axons were clustered in bundles and infiltrated by small round lymphocytes. By argentaffin staining, regenerated axons grew in tissues of injured region, which mostly accompanied with fascicular-arranged multipolar cells. In the control group, spinal cord became thin markedly and spinal meninges were integrated. No new axon appeared in the injured spinal cord by HE staining. No regenerated axon appeared by argentaffin staining, either. ②In the cell transplantation group, most multipolar cells were clustered in bundles. A mass of positive granules of nerve growth factor receptor antibody appeared in cytoplasm, which further verified that OECs still survived and integrated well within the host 6 weeks after transplantation. Linear MBP positive fibers appeared in the injured region by MBP staining,which indicated that myelin-like substance appeared and drew closely in both ends of injured region. At the same time,MBP positive substance also appeared in the multipolar cells, which illustrated that transplanted OECs could induce the occurrence of myelin-like substance. No regenerated axon was found in the control group.CONCLUSION: Delayed transplantation of OECs can survive and induce the occurrence of myelin-like substance in injured spinal cord of adult rats.

Human bone marrow-derived mesenchymal stem cells differentiate into chondrocytes in vitro / 中国组织工程研究

BACKGROUND:How to obtain sufficient autogeneic chondrocytes is a problem which must be answered as soon as possible in both the transplantation of chondrocytes and the development of cartilage engineering.Bone marrow-derived mesenchymal stem cells have the potential of multidirectional differentiation. Under different induced conditions, they can differentiate into multiple tissue cells, such as chondrocyte, osteoblasts , sarcoblast, nerve cells and so on. Insulin-like growth factor-I plays an important role in regulating the formation and development of limb and cartilage.OBJECTIVE:To observe the effect of the insulin-like growth factor-I and culture solution of chondrocyte on inducing bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes in vitro.DESIGN:Open experiment.MATERIALS:This experiment was carried out at the Medical Study Center, Second Hospital Affiliated to Sun Yat-sen University from March to November 2003.Human bone marrow-derived mesenchymal stem cells were harvested from 4 to 6-month old embryos, all of which were from pregnant women who needed to terminate of pregnancy by induction delivery with water bag for health.METHODS:Human embryonic mesenchymal stem cells were isolated with Percoll separating medium. Subsequently, the cells were amplified in vitro,and the expression of surface makers of bone marrow-derived mesenchymal stem cells, such as CD44, CD71, CD34 and CD45 were measured with flow cytometer to identify the cells in our experiment. 100 μg/L insulin-like growth factors and culture solution of chondrocytes were added in the culture medium of bone marrow-derived mesenchymal stem cells of the fourth generation. The morphological changes of induced cells were observed with an inverted microscope. The expression of type Ⅱ cartilage matrix was observed by collogen immunohistochemistry. The proteoglycan level in the cells was detected, too.MAIN OUTCOME MEASURES:Phenotype of bone marrow-derived mesenchymal stem cells was identified through detecting the expressions of CD34, CD44 and CD4.Type Ⅱ collagen immunohistochemistry and the change of cellular ability to secrete proteoglycan after induction were observed to determine whether bone marrow-derived mesenchymal stem cells can differentiate into chondrocytes.RESULTS: ① Being observed under an inverted microscope,the bone marrow-derived mesenchymal stem cells presented a morphology like fibroblasts when they were cultured in vitro. ②Identification of surface antigen of bone marrow-derived mesenchymal stem cells: These cells were detected to have good homogenicity with flow cytometer. The fourth generation of bone marrow-derived stem cells positively expressed CD44, negatively expressed CD34 and CD45, suggesting these cells had the characteristics of bone marrow-derived mesenchymal stem cells. ③Observation of the morphological change of chondrocytes induced by bone marrow-derived mesenchymal stem cells under optical microscope: chondrocyte condition culture solution and insulin-like growth factors-Ⅰ were added to co-culture.During the process of culture, bone marrow-derived mesenchymal stem cells were seen to have a shape of round gradually. Fifteen days later, some cells presented a shape of short-shuttle or polygon with short mutations,which were the shape characteristics of chondrocytes. ④Immunohistochemical staining of Type Ⅱ collagen: In the insulin-like growth factor group,72.5% cells had many brown granules in cytoplasm, which were weakly positive or strongly positive expression of Type Ⅱ collagen. In the control group, bone marrow-derived mesenchymal stem cells was negative expression of type Ⅱ collagen on the 15th day. ⑤ Measurement of proteoglycan level: After co-culture with insulin-like growth factor-Ⅰ and chondrocyte culture solution for 15 days, proteoglycan was higher in the cells of co-culture group [ (8.92±0.91) μg/L ] than in culture group [(2.56±0.26) μg/L,P ＜ 0.05], but lower than in the chondrocyte group[(13.69±1.51) μg/L, P＜ 0.05].CONCLUSION:Insulin-like growth factor-Ⅰ and chondrocyte culture solution can induce bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes.

Biological characteristics of olfactory ensheathing cells after transplantation into spine cord / 中国组织工程研究

BACKGROUND:Olfactory ensheathing cells (OECs) have been shown to possess the potential of repairing injured spinal cord, but their biological characteristics after transplantation in vivo are not well understood.OBJECTIVE: To investigate the migration of OECs after transplantation into the injured spinal cord of adult rats.DESIGN: Randomized and controlled experiment.SETTING: Department of Orthopedics, Guangdong Provincial People's Hospital; Experimental Animal Center of the North Campus of Sun Yat-sen UniversityMATERIALS: Totally 38 2-month-old male SD rats with body mass of (350 ±20) g were used in this study.METHODS: This experiment was conducted in the Experimental Animal Center, North Campus of Sun Yat-sen University between February 2004and May 2004. Two SD rats were used to extract the OECs, which were stained with Hoechst 33342. Totally 36 SD rats were subsequently randomized into 3 groups, namely rostral transplantation group, caudal transplantation group and control group with 12 rats in each group. The rats in the rostral and caudal transplantation groups subjected to operations to establish thoracic spinal cord injury model and OEC suspension was injected; in the control group, the rats were spared of thoracic spinal cord injury with only OEC suspension injection.MAIN OUTCOME MEASURES: Distribution of OECs in the spinal cord was observed under fluorescence microscope 1, 2, 4 and 6 weeks after operation, respectively.RESULTS: Of the rats in the 3 groups, 1 died in the rostral group, and 2in each of the caudal transplantation and control groups, leaving 29 rats for result analysis. The OECs in the rostral and caudal transplantation groups migrated longitudinally along the long axis of the spinal cord to a farthest distance of 8 mm and penetrating the scar tissues, but very few cells could reach the contralateral side. The OECs in the control group diffused locally without migration.CONCLUSION: OECs mainly migrate along the axons in white matter of the injured spinal cord, and their rostral and caudal migration does not differ in speed or amount. Only a small amout of OECs can across the transected gap of the spinal cord.

Influences of olfactory ensheathing cells transplantation on axonal regeneration in spinal cord of adult rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe whether olfactory ensheathing cells could be used to promote axonal regeneration in a spontaneously nonregenerating system.</p><p><b>METHODS</b>After laminectomy at the lower thoracic level, the spinal cords of adult rats were exposed and completely transected at T10. A suspension of ensheathing cells was injected into the lesion site in 12 adult rats, and control D/F-12 (1:1 mixture of DMEM and Ham's F-12) was injected in 12 adult rats. Six weeks and ten weeks after cell transplantation, the rats were evaluated by climbing test and motor evoked potentials (MEPs) monitoring. The samples were procured and studied with histologicl and immunohistochemical methods.</p><p><b>RESULTS</b>At the 6th week after cell transplantation, all the rats in both the transplanted and control groups were paraplegic and the MEPs could not be recorded. At the 10th week after cell transplantation, of 7 rats in the control group, 2 rats had muscles' contraction of the lower extremities, 2 rats had hips and/or knees' active movement; and 5 rats' MEPs could be recorded in the hind limbs in the transplanted group (n=7). None of the rats in the control group had functional improvement and no MEPs recorded (n=7). Numerous regenerating axons were observed through the transplantation and continued to regenerate into the denervated host tract. Cell labelling using anti-Myelin Basic Protein (MBP) and anti-Nerve Growth Factor Receptor (anti-NGFR) indicated that the regenerated axons were derived from the appropriate neuronal source and that donor cells migrated into the denervated host tract. But axonal degeneration existed and regenerating axons were not observed within the spinal cords of the adult rats with only D/F-12 injection.</p><p><b>CONCLUSIONS</b>The axonal regeneration in the transected adult rat spinal cord is possible after ensheathing cells transplantation.</p>

In situ detection and significance of hTERT in giant cell tumor of bone / 中国病理生理杂志

AIM: human telomerase reverse transcriptase (hTERT) is the catalytic subunit of telomerase, and hTERT mRNA expression contributes to telomerase activity. We therefore assessed the significance of hTERT expression in the biological behavior of giant cell tumor(GCT) of bone. METHODS: In situ hybridization was used to identify the expression of hTERT mRNA in 27 formalin-fixed paraffin-embedded clinical specimens from human GCT. RESULTS: 8 cases (8/27)displayed positive hTERT mRNA signal, positive hTERT mRNA signal located in the cytoplasms of mononuclear stromal cells and some multinuclear giant cells. CONCLUSION: The expression of telomerase in paraffin-embedded tissue can be evaluated efficiently and effectively by in situ hybridization, and the results suggest that there may be a neoplastic multinuclear giant cell type in human GCT.

The influence of transplantation of cryopreserved olfactory ensheathingc ells on spinal cord axon regeneration in adult rats / 中华骨科杂志

Objective To observe the effects of cryopreserved olfactory ensheathing cells(OECs )transplantation on spinal cord axonal regeneration and spinal func tional improvement.Methods Twenty-four rats were divided into experimental a nd control groups,with each group of 12rats.The spinal cord in-jury was es tablished by transecting the spinal cord with microsurgery scissors.OECs were p urified from SD rat olfactory bulb and cultured in DMEM and cryopreserved(-1 20℃)for 2weeks.OECs suspension[(1～1.4)?10 5 /ml ]was transplanted into transected spinal cord at T 10 level,while the DMEM solution was in ject-ed instead in the control group .At 6weeks and12weeks after transplantation,the rats were evaluated with cl imbing test and MEP monitoring.The samples of spinal cord were procured and st udied with histological and immunohistochemical measurement.Results At 6weeks after transplantation,all of the rats in both transplanted and control group were paraplegic,and MEPs could not be recorded.Morphol ogy of trans-plante d OECs was normal,and OECs were interfused with host well.Axons could regrow i nto gap tis sue be-tween the spinal cords.Both OECs and regrowed axons were im munoreactive for MBP.No re grown axons were found in the control group.At 12weeks after transplantation,2rats(2/7)had lower ex tremities mus-cle contraction,2rats(2/7)had hip and /or knee active movement,and MEP of 5rats(5/7)could be recorded in the calf in the transplantation group.Non e of the rates(7/7)in the control group had functional improvement,and n one of them had MEPs recorded.In the transplanted group,histological and imm unohis-tochemical methods showed the number of transplanted OECs reduced and s ome regrown axons had reached the end of transected spinal cord.However,no r egrown axons could be seen except scar forma tion in the control group.Concl usion Cryopreserved OECs could integrate with the host and promote re growing a x-ons across the transected spinal cord ends.