N-Butanol Extract of Modified You-Gui-Yin Attenuates Osteoclastogenesis and Ameliorates Osteoporosis by Inhibiting RANKL-Mediated NF-κB Signaling.

Postmenopausal Osteoporosis (PMOP) is the most prevalent primary osteoporosis, attributable to an imbalance in osteoblast and osteoclast activity. Modified You-Gui-Yin (MYGY), a traditional Chinese herbal formula, is able to effectively treat PMOP, while the critical components and pharmacological mechanisms of MYGY are still unclear. In this study, we aimed to investigate the therapeutic effects and underlying mechanisms of N-butanol extract of MYGY (MYGY-Nb) in ovariectomized (OVX)-induced osteoporosis mice. Histological staining and micro-computed tomography (µCT) analysis showed that MYGY-Nb was more effective in the suppression of OVX-induced bone loss than MYGY original formula. Subsequently, liquid chromatography and mass spectrometry analysis identified 16 critical compounds of MYGY-Nb and some of them are reported to affect osteoclast functions. Furthermore, in vivo and in vitro experiments demonstrated that MYGY-Nb significantly attenuated osteoclastogenesis by down-regulating RANKL-mediated NF-κB signaling. In conclusion, our study indicated that MYGY-Nb suppresses NF-κB signaling and osteoclast formation to mitigate bone loss in PMOP, implying that MYGY-Nb and its compounds are potential candidates for development of anti-PMOP drugs.

A Hyperglycemic Microenvironment Inhibits Tendon-to-Bone Healing through the let-7b-5p/CFTR Pathway.

BACKGROUND: Tendon-to-bone healing is a difficult process in treatment of rotator cuff tear (RCT). In addition, diabetes is an important risk factor for poor tendon-to-bone healing. Therefore, we investigated the specific mechanisms through which diabetes affects tendon-to-bone healing by regulating the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). METHODS: Tendon-derived stem cells (TDSCs) were extracted from rats after which their proliferative capacities were evaluated by the MTT assay. The expression levels of CFTR and tendon-related markers were determined by qRT-PCR. Then, bioinformatics analyses and dual luciferase reporter gene assays were used to identify miRNAs with the ability to bind CFTR mRNA. Finally, CFTR was overexpressed in TDSCs to validate the specific mechanisms through which the high glucose microenvironment inhibits tendon-to-bone healing. RESULTS: The high glucose microenvironment downregulated mRNA expression levels of tendon-related markers and CFTR in TDSCs cultured with different glucose concentrations. Additionally, bioinformatics analyses revealed that let-7b-5p may be regulated by the high glucose microenvironment and can regulate CFTR levels. Moreover, a dual luciferase reporter gene assay was used to confirm that let-7b-5p targets and binds CFTR mRNA. Additional experiments also confirmed that overexpressed CFTR effectively reversed the negative effects of the hyperglycaemic microenvironment and upregulation of let-7b-5p on TDSC proliferation and differentiation. These findings imply that the hyperglycemic microenvironment inhibits CFTR transcription and, consequently, proliferation and differentiation of TDSCs in vitro by upregulating let-7b-5p. CONCLUSIONS: A hyperglycemic microenvironment inhibits TDSC proliferation in vitro via the let-7b-5p/CFTR pathway, and this is a potential mechanism in diabetes-induced poor tendon-to-bone healing.

Urine Metabolomics Profiling of Lumbar Disc Herniation and its Traditional Chinese Medicine Subtypes in Patients Through Gas Chromatography Coupled With Mass Spectrometry.

Lumbar disc herniation (LDH) possesses complex pathogenesis, which has not been well elucidated yet. To date, specific or early diagnosis of LDH remains unavailable, resulting in missed opportunity for effective treatment. According to Traditional Chinese medicine (TCM) theory, LDH can be divided into two subtypes (reality syndrome and deficiency syndrome). The purpose of this study was to analyze the metabolic disorders of LDH and its TCM subtypes and screen out potential biomarkers for LDH diagnosis. Gas chromatography coupled with mass spectrometry (GC-MS) was applied to test the urine samples from 66 participants (30 healthy volunteers, 18 LDH patients with deficiency syndrome and 18 patients with reality syndrome). PCA analysis showed a distinct separation tendency between the healthy subjects and LDH patients but no obvious separation between the different syndromes (reality syndrome and deficiency syndrome) of LDH patients. As a result, 23 metabolites were identified significantly altered in the LDH patients, as compared with the healthy subjects. The altered metabolites belong to amino acid metabolism, nucleic acid metabolism, carbohydrate metabolism, and vitamin metabolism, which are related to osteoporosis and inflammation. Our results indicate metabolic disorders of LDH and thereby propose a group of metabolic biomarkers for potential application in early diagnosis of LDH in clinic, which provide a reasonable explanation for the pathogenesis of LDH.

Identification of the specific microRNAs and competitive endogenous RNA mechanisms in osteoporosis.

OBJECTIVE: Osteoporosis and osteoarthritis are metabolic skeletal disorders. This study aimed to identify specific networks of competitive endogenous RNA (ceRNA) in osteoporosis that differ from those in osteoarthritis. METHODS: The dataset GSE74209 was downloaded from the Gene Expression Omnibus, and differentially expressed microRNAs (DEmiRNAs) in osteoporotic samples and osteoarthritic samples were identified. After predicting target genes and linked long noncoding (lnc)RNAs, ceRNA networks of DEmiRNAs were constructed. The nodes that overlapped between ceRNA networks and the Comparative Toxicogenomics Database were selected as key candidates. RESULTS: Fifteen DEmiRNAs (including 2 downregulated and 13 upregulated miRNAs) were identified in osteoporotic samples versus osteoarthritic samples; these targeted 161 genes and linked to 60 lncRNAs. The ceRNA network consisted of 6 DEmiRNAs, 63 target genes, and 53 lncRNAs. After searching the Comparative Toxicogenomics Database and mining the literature, 2 lncRNAs (MALAT1 and NEAT1), 2 DEmiRNAs (hsa-miR-32-3p, downregulated; and hsa-miR-22-3p, upregulated) and 6 genes (SP1, PTEN, ESR1, ERBB3, CSF1R, and CDK6) that relate to cell death, growth, and differentiation were identified as key candidates separating osteoporosis from osteoarthritis. CONCLUSIONS: Two miRNA-ceRNA networks (including NEAT1/MALAT1-hsa-miR-32-3p-SP1/FZD6 and NEAT1/MALAT1-hsa-miR-22-3p-PTEN/ESR1/ERBB3/CSF1R/CDK6) might have crucial and specific roles in osteoporosis.

Reprogramming rat embryonic fibroblasts into induced pluripotent stem cells using transposon vectors and their chondrogenic differentiation in vitro.

It has been known that the successful reprogramming of differentiated human somatic cells into a pluripotent state would allow for the creation of cartilage cells. However, current virus­mediated strategies to form induced pluripotent stem cells (iPSCs) are limited in their clinical application due to exogenous gene modification. In the present study, the piggyBac transposon system carrying corresponding genes (Oct3/4, Klf4, Sox2 and c­Myc) was employed to reprogram rat embryonic fibroblasts (REFs) into iPSCs, and the transposon­carried genes were successfully removed by a transposase system. Furthermore, the differentiation capacity of the iPSCs into chondrocytes was investigated in vitro. A typical chondrocyte marker, collagen­II, was expressed following culture. In conclusion, rat iPSCs without genetic modification were obtained from REFs using the piggyBac transposon system and the transposase mPB system, and these cells possessed the capacity of chondrocyte differentiation, suggesting that this method may be an effective approach for the treatment of cartilage disorders in the future.

[Treatment of non-traumatic femoral head avascular necrosis by perfusion of bone marrow stromal stem cells through optional artery].

OBJECTIVE: To study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis. METHODS: From January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis. RESULTS: The patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening. CONCLUSION: Perfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.

Plasma metabonomic profiling of lumbar disc herniation and its traditional Chinese medicine subtypes in patients by using gas chromatography coupled with mass spectrometry.

Lumbar disc herniation (LDH) is a commonly occurring disease, threatening human health and life quality. Lack of a gold standard of diagnosis has hindered the efficiency and efficacy of clinical therapy against LDH. Traditional Chinese medicine (TCM) has provided an experience-based but subjective diagnosis system for LDH, demanding objective evidence and explanation. In this study, we adopted a metabonomics approach using gas chromatography-mass spectrometry (GC-MS) to profile metabolic characteristics of LDH and its TCM subtypes. Plasma samples of 41 LDH patients and 25 healthy controls were collected. LDH patients were classified into two main subtypes, the reality syndrome and deficiency syndrome, according to TCM theory. By using multivariate statistical analysis and metabolism network analysis, we found diverse perturbations of metabolites in amino acid metabolism and carbohydrate metabolism, in which the amino acids (glutamic acid, aspartic acid, glycine, etc.) were up-regulated and a key carbohydrate metabolite (glucose 1-phosphate) was down-regulated. Few differences were found between the two TCM subtypes. Our findings reveal the metabolic disorders of LDH for the first time and demonstrate the feasibility of the metabonomics approach for LDH research but not for its TCM subtypes.

[Case-control study on therapeutic effects between modified Weaver-Dunn surgery and clavicular hook plate fixation in the treatment of acromioclavicular joint dislocation].

OBJECTIVE: To study therapeutic effects between hook plate fixation and modified Weaver-Dunn surgery for the treatment of acromioclavicular joint dislocation. METHODS: Forty patients with fresh acromioclavicular joint dislocations of type III according to Rockwood classification were reviewed. All the patients were divided into two groups: hook plate fixation group and modified surgery group. There were 20 patients in hook plate fixation group, including 13 males and 7 females, with an average age of (37.45 +/- 14.29) years old; 12 patients had injuries in the left and 8 patients had injuries in the right; preoperative Constant-Murley score was 40.75 +/- 10.40. And there were 20 patients in modified surgery group,including 11 males and 9 females, with an average age of (41.65 +/- 14.83) years old; 11 patients had injuries in the left and 9 patients had injuries in the right; preoperative Constant-Murley score was 42.75 +/- 8.18. The Lazzcano standard, Constant-Murley score and imaging changes were used to evaluate shoulder joint function before and after surgery. RESULTS: All the patients were followed up,and the duration ranged from 7 to 32 months,with an average of 24 months. According to Lazzcano evaluation, 16 patients got an excellent result,3 good and 1 poor in modified surgery group with no re-dislocation, and 1 patient had pain more than middle degree; while in hook plate fixation group, 9 patients got an excellent result, 7 good and 4 poor, 1 patient had re-dislocation, and 3 patients got pain more than middle degree. The therapeutic effects of modified surgery group were better than those of hook plate fixation group. Constant-Murley scores:preoperative 42.75 +/- 8.18 vs 93.40 +/- 4.04 at the latest follow-up in modified surgery group; preoperative 40.75 +/- 10.40 vs postoperative 88.40 +/- 4.81 and 92.05 +/- 4.49 at the latest follow-up in hook plate fixation group. The postoperative scores all improved compared to preoperative scores in two groups. And there was no statistical difference of scores at the latest follow-up between two groups. CONCLUSION: The surgery of allograft tendon transplantation combined with anchor fixation to strengthen coracoclavicular ligament, as well as part transposition of acromiocoracoid ligament and resection at the distal part of clavicle may got safety fixation and less postoperative complications compared with hook plate internal fixation.

[Short-term curative effects of Tantalum rod treatment in early avascular necrosis].

OBJECTIVE: To explore the recent clinical curative effect of Tantalum rod in treating the early avascular necrosis. METHODS: From January 2008 to November 2008, the 25 patients (39 hips) with early avascular necrosis accepted tantalum rod placement and included 9 males (11 hips) and 16 females (28 hips) with an average age of 37 years old ranging from 18 to 74 years old. Four patients (6 hips) caused by Alcoholic, 6 patients (8 hips) by hormone, 2 cases (2 hips) by traumatic, 13 cases (23 hips) by idiopathic. Steinberg preoperative stage involved 7 hips in period I, 24 hips in period II, 8 hips in period III. Curative effect analysis included preoperative and postoperative Harris score, radiographic changes and hip replacement for follow-up to accept the end of the femoral head survival rate. RESULTS: All patients were followed up for 6 to 47 months (averaged 37.4 months). All 12 hips imaging appeard progress,including tantalum rod exit in 1 hip, hip hemiarthroplasty collapse in 3 hips, the area increased to avascular necrosis in 8 hips. Six hips accepted total hip replacement, including imaging progress in 5 hips (41.7%, 5/12), no imaging progress in 1 hip (3.7%,1/27). All hips' Kaplan-Meier survival curves showed 6-month survival rate was (97.4 +/- 2.5)% after tantalum stick insertion, 1-year survival rate was (94.7 +/- 3.6), and 2-year survival rate was (88.6 +/- 5.4)%, 3-year survival rate was (72.5 +/- 11.2). CONCLUSION: It is effective for treatment of avascular necrosis of femoral head in Steinberg I and II by Tantalum rod, and it can effectively relieve femoral head replacement time.

[Progress on tantalum rod implanting for the treatment of femur head necrosis].

Incorrect treatment for femur head necrosis can cause collapse of femoral head and tresult in severe harm for the patients (especially for the patient with middle-aged and young). The structure and mechanics characteristics of tantalum rod is similar to bone tissue, it higher strength and can adapt the internal environment of organism, so it has a large potency in treating femur head necrosis. Treatment of early femur head necrosis with tantalum rod implanting had alreadly widey applied at home and abroad, the method has the advantages of simple operation, little risk, less complication and beseems the patient with stage I - II of ARCO. But reasons that the difficult diagnosis of early femur head necrosis, localized effect of tantalum rod, different experience of medical worker,caused the contentions about effect of tantalum rod implanting. With development of science, tantalum rod implanting combined with correlative biotechnology should raise the effect in treating femur head necrosis.