Role of ATP6V1H gene in bone metabolism / 中国骨伤

Osteoporosis is one of the common clinical orthopedic diseases, which can lead to a variety of complications. There are many pathogenic factors in this disease. The latest research found that ATP6V1H is a new gene leading to the occurrence of osteoporosis, and it is likely to become a new target for the future drug treatment of osteoporosis.This paper introduces the biological structure and characteristics of H subunit, summed up the human body caused by loss of ATP6V1H and animal models such as zebrafish, mice bone loss and osteoporosis symptom such as related research reports of the loss, from osteoclast, osteoblast and marrow stromal cell level and the connection between the various subunits further expounds the H subunit regulate bone dynamic balance of mechanism, to explore ATP6V1H in bone developmentand bone related diseases has laid a solid foundation, also provide new ideas for clinical treatment of osteoporosis.

Research status of correlation between type IX collagen gene and musculoskeletal diseases / 中国骨伤

As an important component of articular cartilage, type IX collagen plays an important role in regulating homeostasis of bone and cartilage. The mutation or deletion of gene could disequilibrate homeostasis leads to deformity of corresponding bone and joint, and finally causes multiple epiphyseal dysplasia. Moreover, anatomic variation also leads to biomechanics change of corresponding functional unit, combine with micro-environment change caused by change of genes, osteoarthritis and disc degeneration were occurred under the function of stress over and over again. In addition, lack of type IX collagen could effect repair of articular cartilage, intervertebral disc tissue injury. However, musculoskeletal diseases related with type IX collagen has so much not limited this, the reports about it is less for lack of evidence, and need further work to study. Clear relationship of type IX collagen and its disease could provide an effective diagnostic method, and develop a new pathway for follow-up treatment.

Application status of 3D printing patient-specific instrumentation in total knee arthroplasty / 中国骨伤

Recently, the joint replacement has gradually become a common operation of orthopedics. It's also the hot spot in current investigations. The increasing number of reports about the 3D printing technology such as patient-specific instrumentation(PSI) is applied to hip and knee replacement. It plays an important role in supporting operation especially for complex cases. However, it is still at the primary stage. This article reviews the relevant literature, analyzes and collates the research status of the application of 3D printing PSI in total knee arthroplasty, and summarizes its effectiveness in the application of knee arthroplasty from the aspects of individual preoperative planning, intraoperative operation and postoperative functional recovery, especially complex knee surgery such as obesity and knee deformity. The necessity of clinical application is discussed, and the existing problems and controversies in the current clinical application are summarized in order to provide references for the future research direction and clinical application.

Essence of "Shen (Kidney) Controlling Bones": Conceptual Analysis Based on Hypothalamic-Pituitary-Adrenal-Osteo-Related Cells Axis / 中国结合医学杂志

As a traditional concept of Chinese medicine (CM), the theory of "Shen (Kidney) controlling bones" has been gradually proven. And in modern allopathic medicine, the multiple mechanisms of bone growth, development and regeneration align with the theory. Shen deficiency as a pathological condition has a negative effect on the skeleton of body, specifically the disorder of bone homeostasis. Present studies indicate that Shen deficiency shares a common disorder characterized by dysfunction of hypothalamic-pituitary-adrenal (HPA) axis. HPA axis may be an important regulator of bone diseases with abnormal homeostasis. Therefore, we posit the existence of hypothalamic-pituitary-adrenal-osteo-related cells axis: cells that comprise bone tissue (osteo-related cells) are targets under the regulation of HPA axis in disorder of bone homeostasis. Chinese herbs for nourishing Shen have potential in the development of treatments for disorder of bone homeostasis.

Subculture, cryopreservation and recovery of osteoclasts / 中国骨伤

<p><b>OBJECTIVE</b>To explore the feasibility of passage, cryopreservation, and recovery of osteoclasts in order to develop new techniques facilitating osteoclast research.</p><p><b>METHODS</b>Passage of osteoclasts: adult male SD rat(SPF grade, weight of 250 g) was sacrificed and the abdominal aorta was exposed for blood draw. Monocytes isolated from peripheral circulation was treated with RANKL and M-CSF for 2 weeks. After formation of osteoclasts, they were trypsinized with pipetting, centrifuged, re-suspended with α-MEM containing RANKL and M-CSF, and cultured in 6 well-plates and 35 mm culture dishes. Freezing of osteoclasts: trypsinized osteoclasts were centrifuged and resuspended with DMSO, FBS, α-MEM (1:2:7), and were stored in liquid nitrogen(-196 °C). Recovery of osteoclasts: frozen osteoclasts were taken out of liquid nitrogen tank and thawed quickly at 37 °C in water bath. After wash with PBS, the cells were resuspended with α-MEM containing RANKL and M-CSF, and were cultured in 6 well dishes and 35 mm culture dishes. Meanwhile, cells were checked with inverted phase contrast microscope and observed in the live cell station for real time imaging. TRAP staining was performed 3 days after plating.</p><p><b>RESULTS</b>Trypsinization together with pipetting and shaking can detach the adherent osteoclasts, and the resuspended cells can be used for passage and storage in liquid nitrogen. The passaged cells became fully attached to the culture dishes in 2 hours, and the multinucleated feature could be clearly seen. The osteoclasts recovered from liquid nitrogen could completely spread out for 2 to 3 hours so that the multinucleated cells were clearly seen. These cells were still TRAP positive.</p><p><b>CONCLUSIONS</b>Although osteoclasts strongly adhere to the bottom of culture dishes, a large majority of the osteoclasts can be detached after appropriate digestion with trypsin, pipetting and shaking. These cells can be used for passage and cryopreservation. After recovering from liquid nitrogen, these cells still preserve the viability and the feature of osteoclasts. The results provide a new and powerful tool for future study of osteoclast biology.</p>

Diagnosis and treatment of esophagustype cervical spondylosis / 中国骨伤

Esophagustype cervical spondylosis is easily misdiagnosed or missed diagnosed for its lower incidence. The pathological basis manily concerned with compression of osteophyte for esophageal and tracheal wall and local inflammatory reaction. The diagnosis of disease is generally not difficult. Especially dynamic esophagogram could not only show degree of cervical degeneration and osteophyte shape, but also could clearly observe location and degree of osteophytosis indenting esophageal and tracheal. Recently, the treatment of esophagustype cervical spondylosis has been the focus of attention. It is generally believed that conservative treatment could alleviate disease but could not cure it. More and more scholars tend to treat disease by surgical resection of osteophyte to relieve compression for esophagus, and clinical symptoms could obviously improved. However, long-term follow up is lack, and further follow-up effect is lack of powerful support, anddifferent treatment methods and surgical procedures are lack of comparative research.

Progress on the early diagnosis of knee osteoarthritis / 中国骨伤

Knee osteoarthritis is one of the common type of arthropathy, the clinical stage of the typical patients belongs to the middle-late stage, so it urges to improve the early diagnosis. At present, magnetic resonance imaging is most used in clinical diagnosis of knee osteoarthritis, and with the development of different MRI sequences, the sequences of early articular cartilage lesions are used in clinic. In the early diagnosis of knee osteoarthritis, the simple and practical methods such as ultrasonography is becoming a trend, and the specific biomarkers of early knee osteoarthritis have become the hot research. This overview article outlined the methods of early diagnosis from the ultrashort echo time MRI, ultrasonography and biomarkers.

Clinical experience on postoperative balance of hemostasis and antithrombus for patients with hemophilic arthritis after arthroplasty / 中国骨伤

<p><b>OBJECTIVE</b>To observe the clinical significance of postoperative personalized antithrombotic therapy for patients with hemophilic arthritis (HA) patients after arthroplasty.</p><p><b>METHODS</b>From September 2005 to October 2013, 11 cases of arthroplasty for hemophilic arthritis in hip and knee total operation 14 times,including 1 case of double knees (calculated as one operation), operation in left knees 6 times, operation in right knees 5 times, 2 in hip. All the patients were male and the age ranged from 23 to 57 years old,with an average of (36.1 ± 11.0) years old; the average weight was (64.1 ± 8.9) kg. All the patients were preoperatively diagnosed and classified as hemophilic arthritis with the radiological images and laboratory tests. According to the function of joints, the risk of postoperative venous thromboembolism (VTE), and dynamic observation of Factor VIII:C (FVIII:C) activity, patients were treated with personalized antithrombus by adjusting the dosage of recombinant human coagulation factor VIII (Kogenate FS). All the patients were orderly divided into postoperatively distal joints moving group and none-moving group to observe the coagulation function.</p><p><b>RESULTS</b>The enrolled patients had no postoperative complication of VTE and pulmonary embolism (PE). The APTT and D-2 were different between two groups in the postoperative early stage. Length of hospital day was shorter in the moving group than none-moving group.</p><p><b>CONCLUSION</b>Because of the self-coagulation disorder, patients with HA tended to bleed. However it doesn't mean that there is no risk of postoperative thrombosis. Therefore,it's important to determine how to control the balance between postoperative antithrombus, hemostasis,and coagulation factor replacement therapy after arthroplasty for HA. Postoperative moving has proved helpful for HA, especially in reducing the risk of hemostasis and shortening the time in hospital.</p>

Clinical diagnosis and treatment of chronic osteomyelitis / 中国骨伤

Chronic osteomyelitis is one of the most common disorder in clinic. In recent years due to diabetes, peripheral vascular disease and trauma induced disease increased, the prevalence rate increased. With the development of magnetic resonance imaging and CT imaging technology, it greatly improved the accuracy of clinical diagnosis of chronic osteomyclitis and ability to describe the infection characteristics, and provide a reliable basis for clinical treatment. The current research on chronic osteomyelitis mainly concentrated on the aspects of imaging applications and ways of using antibiotic optimization control inflammation, defect restoration and reconstruction of blood supply and treatment. But the best time to the antibiotic therapy and the use of program is still uncertain, for after debridement, bone grafting time and defect repair function of fast recovery still need further research.

Effects of serum of Bushen Huoxue prescription (Chinese characters) on classic Wnt/β-catenin signaling pathways of osteoblasts / 中国骨伤

<p><b>OBJECTIVE</b>To observe effects of serum Bushen Huoxue prescription(Chinese characters) on classic Wnt/β-catenin signaling pathways of osteoblasts, and explore mechanism of Bushen Huoxue prescription (Chinese characters) for preventing osteoporosis.</p><p><b>METHODS</b>Twenty health female rats were randomly divided into two groups, including Bushen Huoxue (Chinese characters) group and saline group,10 in each group. Bushen Huoxue (Chinese characters) group and Saline group were gavaged Bushen Huoxue and saline every day for 1 week. Bushenhuoxue containing serum and saline containing serum were got according to methods of serum preparation of drug-containing. The osteoblasts was cultured with neonatal rat skull according to Enzyme Consumer Law, and was identified by Wright-Giemsa staining (R-J) and alkaline phosphatase staining (ALP). The third generation of osteoblasts was divided into three groups, including saline group, normal group,Bushen Huoxue (Chinese characters) group. Each group were added to 15% appropriate medium. ALP activity of osteoblasts and osteoblasts proliferation rate were tested, mineralized nodules were observed, the expression of β-catenin, Runx2, Osx mRNA of osteoblasts were tested by RT-PCR.</p><p><b>RESULTS</b>There was blue granules in cytoplasm, cell nucleus was flint with 1 to 3 nucleoli showed by R-J staining, morphology of osteoblasts were cultured. ALP staining showed cytoplasm with purple granules, the results showed that the cultured cell was osteoblasts. The content of ALP in Bushen Huoxue (Chinese characters)group was (6.272±0.131) ,appreciation rate was (0.81? 0.172), and could significantly improve differentiation and proliferation activity of osteoblasts compared with Saline group (P< 0.01). There were four different size orange nodules, the Maximun nodule was 1.0 x 1.0 cm in Bushen Huoxue (Chinese characters) group after Alizarin red staining, the results showed Bushen Huoxue (Chinese characters)group could obviously improve mineralization of osteoblasts. The expression of mRNA of β-catenin, Runx2 and Osx in Bushen Huoxue (Chinese characters)group were (1.782±0.944), (1.935±0.994) and (1.610±0.811) by RT-PCR,it was significantly increased compared with saline group (P<0.01), but there was no difference between Bushen Huoxue (Chinese characters)group and normal group (P>0.05).</p><p><b>CONCLUSION</b>Bushen Huoxue (Chinese characters)group could obviously promote differentiation, proliferation and mineralization of osteoblasts through activation of Wnt, β-catenin signaling pathway. It suggested that the mechanism of action of Bushen Huoxue (O'f f Il.t)particle clould prevent osteoporosis through the activation of Wnt, β-catenin signaling pathway.</p>

Research on regulation mechanism of osteoclast differentiation / 中国骨伤

Osteoclasts are multinucleated giant cell, which derived from mononuclear myeloid hematopoietic stem cells with the function of bone absorption. Osteoclasts plays a key role in bone metabolism, therefore the body is very strict to regulation of osteoclastogenesis. Mobilization and differentiation of osteoclast maturation is a complex and sophisticated multi-level regulatory processes. In the relevant regulatory mechanisms, OPG/RANKL/RANK system plays a pivotal role in the process of osteoclast differentiation and maturation. Recent studies revealed that immune cells and osteoclasts were closely connect with each other in the field of bone metabolism, also provide a new therapeutic target for the treatment of bone diseases. The apoptosis of osteoclasts in bone metabolism have been payed more attention,while its mechanism is still not clear, which need further research.

Research advance on intervertebral disc degeneration and cell death / 中国骨伤

Intervertebral disc degeneration is considered as a primary cause of clinical low back pain, however the molecular mechanism is not clear yet. Recently, researches on the molecular basis of intervertebral disc degeneration have become a hotspot. The special structure and biomechanics properties of the disc contribute to its propensity toward degeneration. Intervertebral disc degeneration is associated with the changes of the cytological behavior,including the increase in cell death and the degradation of extracellular matrix. However, the mechanism of cell death including cell apoptosis and autophagy in intervertebral disc degeneration remains unclear. Further study on the molecular mechanism of intervertebral disc degeneration is the foundation of improving and treating the intervertebral disc degeneration in the future. Although some progresses are made in the aspect of biological study, the biological environment of intervertebral disc itself is still a challenge for the development of biological treatment. This article is to review the latest advance on the biological characteristics of normal intervertebral disc and the cell death in the process of the intervertebral disc degeneration.

Research on application of determination of MMP-13 in osteoarthritis / 中国骨伤

Osteoarthritis (OA) is a complex chronic progressive disease attacked by biological and mechanical factors and a result from the anabolic and catabolic imbalance in chondrocyte, subchondral bone and extracellular matrix(ECM). Etiology and pathological of OA are not yet entirely clear. The degradation and destruction of collagen II caused by matrix metalloproteinase -13 (MMP-13) is considered the core factor in the occurrence and development of OA. The research of MMP-13 inhibitor provide ideas and methods for the treatment of OA. In this article,the role and determination of MMP-13 in OA and the development prospect of MMP-13 inhibitor in the treatment of OA research progress were reviewed.

Extracapsular arthroscopic excision of popliteal cysts through anterior combined with posterior approach / 中国骨伤

<p><b>OBJECTIVE</b>To study the clinical effect of arthroscopic excision of popliteal cysts through a combined anterior and posterior approach.</p><p><b>METHODS</b>From January 2010 to December 2012,20 patients with popliteal cysts were treated with arthroscopic excision. There were 14 males and 6 females, with an average age of 49.5 years old, ranging from 45 to 65 years old. The lump has been found for 4 to 18 months,with a mean time of 12 months. Their mean sagittal diameter was 4.5 cm (ranged from 3 to 7 cm). There were 12 popliteal cysts in the left and 8 popliteal cysts in the right. The main clinical manifestation included lump at popliteal fossa, swelling and pain at knee joint and some extent of dysfunction. All diagnoses were determined according to MRI, which clearly showed the communication of cyst and articular cavity. The cyst was removed under arthroscopy, through the posterior approach and then the intra-articular lesion was treated via the anterior approach. According to Rauschning and Lindgren classification, 2 patients were grade I, 6 patients were grade II, and 12 patients were grade III. The guidelines of Rauschning and Lindgren were used to evaluate the clinical effects.</p><p><b>RESULTS</b>No complications such as the injury of blood vessel and nerve, or incision infection occurred in all patients. All the patients were followed up, and the duration ranged from 8 to 24 months, with a mean time of 16 months. According to the criteria of Rauschning and Lindgren, there were 14 cases of grade 0, and 6 cases of grade I after operation, which was improved obviously compared with that pre-operation. No cyst reoccurred and the knee joint pain was relieved.</p><p><b>CONCLUSION</b>Treatment of popliteal cysts with arthroscopic excision through a combined anterior and posterior approach is effective to remove the cyst sac and treat intra-articular diseases simultaneously, resulting in the decrease recurrence rate of cyst.</p>

Methods and evaluations on the sterioid-induced osteoporosis mice model with the type of Kidney-Yin deficiency / 中国骨伤

<p><b>OBJECTIVE</b>To establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.</p><p><b>METHODS</b>Totally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.</p><p><b>RESULTS</b>(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.</p><p><b>CONCLUSION</b>the model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.</p>

Different biological characteristics between nucleus pulposus and annulus fibrosus cells in rabbits / 中国骨伤

<p><b>OBJECTIVE</b>To compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model.</p><p><b>METHODS</b>Five New Zealand white rabbits (2 to 3 kg, either gender) were isolated nucleus pulposus and annulus fibrosus under sterilized condition, then cultured in nutrient solution with 15% FBS and DMEM/F12 (1:1) by enzyme digestion combined with tissue block method. When 90% cells fused, subcultring were performed. Cell morphology were observed by inverted phase contrast microscope, cell viability were detected by trypan blue staining, histological were observed by a toluidine blue and HE staining, cell proliferation were tested by MTT method, then the cell morphology, viability, proliferation between nucleus pulposus and annulus fibrosus were compared.</p><p><b>RESULTS</b>There were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later, and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day.</p><p><b>CONCLUSION</b>The cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus,local biomechnical changes, furether caused structure change and function loss of annulus fibrosus.</p>

Research on osteoporosis model rats with kidney deficiency syndrome / 中国骨伤

<p><b>OBJECTIVE</b>To observe general conditions, physiological condition, viscera wet weight and kidney asthenia serological index in osteoporosis model rats established by bilateral ovarian resection, and provide basis for treating osteoporosis by invigorating kidney.</p><p><b>METHODS</b>Twenty-four female SD rats were randomly divided into 3 groups as model group, sham group and normal group. After anesthesia,model group were treated by bilateral ovarian resection, sham group by sham operation, and normal group without any processing. Pathological tissue and bone mineral density (BMD) of femur was performed 3 months postoperatively. The general conditions and physiological conditions were processed after successfully modeling. Electronic balance was used to collect the weight of kidney, drenal, pituitary, and uterus. Estrogen (E2), Adrenocorticotropic hormone (ACTH), Three typical thyroid original acid (T3), Four typical thyroid original acid (T4) were detected by ELISA.</p><p><b>RESULTS</b>Osteoporosis model appeared with syndromes of kidney deficiency, such as body hair without burnish, chills, cluster, sluggishness, eyes dark, eye movement slow, slow to respond to the delay, decreased appetite, activities slow, body fat, and drowsiness, defecate slightly thin andclear abundant urine. Mobility, anal temperature and food-intake were lower than normal and sham group; the level of ACTH, E2, T3, T4 lower than sham and normal group; the weight of kidney, drenal, pituitary, and uterus also lower than other two groups.</p><p><b>CONCLUSION</b>Osteoporotic model rats, established by bilateral ovarian resection, have manifestation of organic and functional performance of kidney deficiency. Therefore, osteoporotic model rats have the pathogenesis of kidney asthenia and clinical manifestations of kidney deficiency.</p>

Study on the effect of Youguiyin () on regulating gene expression profile of the rats with femoral head necrosis induced by steroid / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the regulation of Youguiyin (YGY, ) on the gene expression profile of the rat with steroid-induced femoral head necrosis (sFHN), for the sake of investigating its molecular mechanism of sFHN prevention and treatment.</p><p><b>METHODS</b>All the 30 rats were randomly divided into three groups, the normal control group (A), the model control group (B), and the YGY treated group (C), 10 in each group. After rats in Groups B and C were being made into FHN models with steroid injection, they received a daily intragastric administration of saline and YGY respectively in equal volume for a total of 6 weeks, while to the unmodeled normal rats in Group A, saline was administered instead. The rats were sacrificed at the terminal of administration; their mRNA from femoral head tissue was extracted and prepared to cDNA probe through inverse transcription for detecting gene expression profile by microarray, outcomes of which was passing fluorescence quantitative PCR verification, and the differential expressed genes were analyzed adopting gene ontology (GO) method.</p><p><b>RESULTS</b>Compared with Group A, the numbers of differential genes found in Groups B and C were 190 and 92, respectively, but the changing trend in the two groups was opposite, mainly manifested as down-regulating in Group B/Group A (GB/GA) and up-regulating in Group C/Group B (GC/GB). The analysis showed that these differential genes were mainly assigned to cell apoptosis, signal transduction, metabolism, cell proliferation and differentiation, cell cycle, blood coagulation, antioxidant activity, etc.</p><p><b>CONCLUSIONS</b>sFHN was regulated by various genes; the regulation of YGY on expressions of these genes and the intra/extra-cellular signaling processes was possibly the molecular mechanism of YGY for preventing/treating sFHN. This study gave an explanation to the effectiveness of Chinese medicine in preventing/treating FHN from aspects of gene expression and enriched the Chinese medicine theory of "Kidney (Shen) governing bones".</p>

Epimedium-derived flavonoids modulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats via Wnt/β-catenin signal pathway activation / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids (EFs) regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis.</p><p><b>METHODS</b>Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- (1 μg/mL), medium- (10 μg/mL) and high- (100 μg/mL) dose EFs were administrated correspondingly. Alkaline phosphatase (ALP) staining, ALP activity determination, oil red O staining and realtime polymerese chain reaction (RT-PCR) were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. Moreover, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 (DKK1) was used in the medium group. Enzymelinked immunosorbent assay (ELISA) and RT-PCR were used to determine mRNA levels of β-catenin, low density lipoprotein receptor-related protein 5 (LRP5) and T cell factor (TCF) protein, known as wnt/β-catenin signal pathway related factors.</p><p><b>RESULTS</b>EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 (Runx2), osteocalcin and collagen I, and decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 (PPARγ-2) and CCAAT enhancer-binding protein-α (C/EBPα) in a dose-dependent manner. While osteoblast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Also EFs up-regulated mRNA expression levels of β-catenin, LRP5 and TCF protein which could be blocked by DKK1.</p><p><b>CONCLUSION</b>EFs regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats by activating wnt/β-catenin signal pathway, which may be an important molecular mechanism of EFs on treating postmenopausal osteoporosis.</p>

Genomics study of steroid induced femoral head necrosis of rats / 中华外科杂志

<p><b>OBJECTIVES</b>To identify significantly differentially expression genes of steroid-induced femoral head necrosis (SINFH) of rats by gene chip, and to find out the potential factors and molecular mechanisms that oxidative stress originate or strengthen the SINFH.</p><p><b>METHODS</b>Twenty Wistar rats were divided into experimental group and control group randomly. E. coli endotoxin was given to all rats at a dose of 20 µg/kg body weight by daily i.p. for two times. Then methylprednisolone (40 mg/kg) or saline was daily injected into the left gluteus muscle of the rats in experimental group and control group respectively. Six weeks later, the mRNA was extracted from the femoral head of rats in every group, and the cDNA were obtained by inverse transcript, then carried out microarray detection. The quantitative RT-PCR was used to confirm the result of microarray, and the differentially expressed genes were analyzed for the functional annotation by gene ontology (GO).</p><p><b>RESULTS</b>Compared to the control group, 190 genes in the experimental group were differentially expressed, with 52 up-regulated and 138 down-regulated. Of these genes, 102 are known (have deposited in GeneBank), while 88 of them are unknown. The known genes can be divided into several families according to their biological functions, such as: oxidative stress, apoptosis, signal transduction, angiogenesis, extracellular matrix, lipid metabolism, and gene transcription related genes. The results of quantitative RT-PCR are consistent with gene-chip results.</p><p><b>CONCLUSIONS</b>The occurrence of SINFH is a complicated process affected by multiple factors and signaling pathways. Our findings indicate that many genes which are involved in different signaling pathways were differentially expressed between SINFH rats and normal rats.</p>