In Hamstring Muscles of Patients With Knee Osteoarthritis an Increased Ultrasound Shear Modulus Indicates a Permanently Elevated Muscle Tonus.

BACKGROUND: Some patients with knee osteoarthritis (KOA) show pain, stiffness and limited flexion and extension at the back of the knee, leading to dysfunction and affecting life. This may be related to changes in the biomechanical properties of skeletal muscles. Shear wave elastography (SWE) can detect these changes by measuring muscle shear modulus. AIMS: To investigate hamstring muscle shear modulus of healthy people and patients was studied using SWE method, and the correlation analysis between the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score of patients' subjective feeling and shear modulus of objective quantification was conducted. METHODS: The hamstring shear modulus was measured by SWE in 50 patients and 50 healthy individuals. Pearson correlation coefficient was used to evaluate the correlation between hamstring stiffness and shear modulus in patients. RESULTS: The hamstring shear modulus were significantly higher in the KOA group [the semimembranosus (SM) 15.23 ± 7.23, the semitendinosus (ST) 15.94 ± 5.40, the biceps femoris long tendinitis (BFL) 14.21 ± 6.55] than in the control group (the SM 10.95 ± 2.41, the ST 11.25 ± 2.23, the BFL 9.98 ± 2.81) (p = 0.000, p = 0.000, p = 0.001). The hamstring shear modulus in the KOA group was moderately positively correlated with pain, shear modulus, and physical function score. CONCLUSION: Preliminary results show that the shear modulus of the hamstring of KOA patients is higher than that of healthy people, the WOMAC score and the shear modulus of patients are moderately correlated. These preliminary results show that ultrasonic shear wave elastography measurement of shear modulus may be enough to sensitive, can detect these effects, more targeted in order to assist the doctor's diagnosis and treatment.

[Poking reduction cannulated screw for the treatment of Sandersâ ¡ calcaneal fracture].

OBJECTIVE: To evaluate clinical effect of poking reduction cannulated screw based on Pingle orthopedic muscle-bone interoperability balance theory in treating Sanders Ⅱ calcaneal fracture. METHODS: From October 2014 to December 2017, 28 patients with Sanders Ⅱ calcaneal fracture were treated with poking reduction cannulated screw guided by Pingle orthopedic muscle-bone interoperability balance theory, including 20 males and 8 females, aged from 24 to 55 years old with an average of (37.2±3.9) years old. Calcaneal width, Bhler angle, and Gissane angle were measured before and after operation, and Maryland Score before and 6 months after operation were compared. RESULTS: All patients were followed up from 12 to 16 months with an average of (13.7±1.3) months. All fractures healed normally, and healing time ranged from 9 to 12 weeks with an average of (10.2±1.3) weeks. No postoperative wound infection, cortical necrosis, or osteomyelitis occurred. The width of the calcaneus decreased from (34.15±2.58) mm before surgery to (30.49±2.37) mm after surgery, Bhler angle increased from (14.16±3.27)° before operation to (31.95±3.07)°after operation, Gissane angle decreased from (128.45±9.04)° before operation to (120.83±8.15)° after operation. Maryland Score was 15.68±4.73 before operation, and was improved to 88.32±2.65 at 6 months after operation;19 patients got excellent result, 6 good, 2 fair and 1 poor. CONCLUSION: Poking reduction cannulated screw based on Pingle orthopedic muscle-bone interoperability balance theory in treating Sanders Ⅱ calcaneal fracture has certain clinical effects, high acceptation of patient, and without special demand for soft tissue around fracture. But it should avoid choosing severe comminuted Sanders Ⅲand Ⅳcalcaneal fracture.

High-Throughput Metabolomics Method for Discovering Metabolic Biomarkers and Pathways to Reveal Effects and Molecular Mechanism of Ethanol Extract From Epimedium Against Osteoporosis.

Metabolomics is an effective strategy to explore the molecular mechanism of herbal medicine. Epimedium, a traditional Chinese herb from the Epimedium brevicornu Maxim., has a therapeutic effect on osteoporosis (OP), however the molecular mechanism of the anti-OP effect is uncle\ar. Therefore, we investigated the pharmacological effect and action mechanism of ethanol extract of epimedium (Ext-epi) onOP rat model. The serum of OP rats was analyzed utilized UPLC-Q-TOF/MS metabolomics, and the potential biomarkers were screened and identified using multivariate data analysis systems and network databases. To further appraise the influence of Ext-epi on biological markers and metabolic pathways, and reveal the potential mechanism of Ext-epi on OP treatment. The results showed that 46 potential biomarkers were screened out and after intervention with Ext-epi extracts solution, 16 potential biomarkers were significantly recalled. Further pathway experiments showed that key pathway analysis include sarachidonic acid metabolism, glycerolphospholipid metabolism as potential targets which is related with the efficacy of Ext-epi protect against OP. These results explain the correlation between metabolites and molecular mechanisms, which is of great significance for understanding the intervention of Ext-epi on OP. In short, based on UPLC-Q-TOF/MS metabolomics may provide effective strategies for understanding the pathogenesis of diseases and evaluating the intervention effect of natural products.

Melatonin inhibits Sirt1-dependent NAMPT and NFAT5 signaling in chondrocytes to attenuate osteoarthritis.

Osteoarthritis (OA) is a degenerative joint disease mainly characterized by cartilage degradation. Interleukin-1ß (IL-1ß) contributes to OA pathogenesis by enhancing oxidative stress and inflammation. Melatonin reportedly elicits potent protection against OA. However, the role of melatonin and underlying mechanism in IL-1ß-stimulated chondrocytes remain largely unclear. In this study, we found that melatonin inhibited IL-1ß-induced toxicity and sirtuin 1 (Sirt1) enhancement in human chondrocytes. Melatonin reduced the IL-1ß-increased nicotinamide phosphoribosyltransferase (NAMPT) expression and the NAD+ level in chondrocytes in a Sirt1-dependent manner. In turn, the inhibitory effect of melatonin on Sirt1 was mediated by NAMPT. Moreover, melatonin suppressed IL-1ß-induced Sirt1-mediated matrix metalloproteinase (MMP)-3 and MMP-13 production. Melatonin also decreased the Sirt1-steered nuclear factor of activated T cells 5 (NFAT5) expression in IL-1ß-challenged chondrocytes. NFAT5 depletion mimicked the suppressive effects of melatonin on IL-1ß-elevated production of inflammatory mediators, including tumor necrosis factor-α (TNF-α), IL-1ß, prostaglandin E2 (PGE2), and nitric oxide (NO) in chondrocytes. TNF-α, IL-1ß, PGE2, or NO decrease caused the similar reduction of MMP-3 and MMP-13 by melatonin in IL-1ß-insulted chondrocytes. Highly consistent with in vitro findings, in vivo results demonstrated that melatonin repressed the expression of relevant genes in rat OA pathogenesis in anterior cruciate ligament transection model. Overall, these results indicate that melatonin effectively reduced IL-1ß-induced MMP production by inhibiting Sirt1-dependent NAMPT and NFAT5 signaling in chondrocytes, suggesting melatonin as a potential therapeutic alternative for chondroprotection of OA patients.

Matrine-induced autophagy counteracts cell apoptosis via the ERK signaling pathway in osteosarcoma cells.

The aim of the present study was to observe whether autophagy was induced by matrine, and to investigate the role of autophagy in the antitumor effects of matrine on human osteosarcoma MG-63 cells and its underlying mechanism. MG-63 cells were cultured in vitro in matrine at a concentration of 0.6, 0.8, 1.0 and 1.2 g/l for 0, 24, 48 and 72 h. A MTT assay was used to evaluate the proliferation inhibition of MG-63 cells by matrine, and Annexin V-fluorescein isothiocyanate/propidum iodide (PI) staining flow cytometry was used to analyze the apoptotic rate. Alterations in cell morphology was assessed by PI and Hoechst 33258 cell staining. Matrine-induced autophagy in MG-63 cells was confirmed by green fluorescent protein-microtubule-associated protein 1-light chain 3 (LC3) b transfection and fluorescence microscopy, and cell viability was investigated by MTT assay following inhibition of autophagy by chloroquine (CQ) pretreatment. The expression level of apoptosis-associated proteins B-cell lymphoma-2 (Bcl-2) and Bcl-2-like protein 4 (Bax), autophagy-associated LC3II protein, and the activation of extracellular signal-regulated kinase (ERK) was detected by western blotting. Cell proliferation was clearly inhibited by matrine in a dose- and time-dependent manner. Flow cytometry and Hoechst 33258/PI staining verified that matrine induced apoptosis in a time-dependent manner when cells were exposed to 1.1 g/l matrine; fluorescence microscopy demonstrated that green fluorescence puncta were enhanced with prolonged time of matrine incubation. Western blotting confirmed that the expression of pro-apoptosis-associated proteins Bax and LC3II, and phosphorylated-ERK were upregulated, and anti-apoptosis protein Bcl-2 was downregulated in a time-dependent manner following treatment with matrine. The cell viability of the matrine + CQ group was increased compared with the matrine group alone, which revealed that matrine treatment alone induced protective autophagy in MG-63 cells. In addiiton, expression of LC3II/LC3I decreased and the expression of BAX/Bcl-2 increased in the matrine + U0126 group compared with the matrine alone group. The present study demonstrated, to the best of our knowledge, for the first time that matrine induced protective autophagy via ERK activation in MG-63 cells, and matrine combined treatment with CQ or U0126 led to an increase in apoptosis in osteosarcoma cells.

Crosstalk between Beclin-1-dependent autophagy and caspase­dependent apoptosis induced by tanshinone IIA in human osteosarcoma MG-63 cells.

The aim of the present study was to ascertain whether or not autophagy is induced by tanshinone IIA (TanIIA), and to explore the crosstalk between autophagy and apoptosis in regards to the antitumor effects of TanIIA on MG-63 cells and the potential mechanism. MG-63 cells were cultured in vitro with various concentrations of TanIIA (0, 2.5, 5, 10 and 20 mg/l) for 0, 24, 48 and 72 h, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to evaluate the inhibition of the proliferation of osteosarcoma MG-63 cells by TanIIA or in the presence/absence of chloroquine (CQ). Autophagic vacuoles and characteristic autophagosomes were observed by transmission electron microscopy (TEM). TanIIA-induced autophagy in MG-63 cells was confirmed by GFP-LC3 punctate fluorescence. The expression levels of apoptosis-related proteins caspase-3, caspase-8, caspase-9 and cleaved-PARP and autophagy-related proteins LC3II/LC3I and Beclin-1 were detected by western blotting. FITC-Annexin V/propidium iodide (PI) staining, flow cytometry and Hoechst 33258 staining were used to analyze the apoptotic rate. Fluorescence intensity of reactive oxygen species (ROS) was examined under a fluorescence microscope using an analysis software system. Cell proliferation was obviously inhibited by TanIIA in a dose- and time-dependent manner. Generation of autophagy was triggered by TanIIA (0-20 mg/l) treatment, and in a Beclin-1-dependent manner. Compared with the control group, the apoptosis ratio following treatment with 2.5 mg/l TanIIA failed to achieve statistical significance. Expression of caspase-3, -8 and -9, and cleaved-PARP in the other groups was gradually enhanced in dose-dependent manner. Our analysis also suggested that the influence of autophagy on TanIIA cytotoxicity had a phase effect; with low­dose drugs and shorter treatment periods, autophagy functioned as a damage repair mechanism. In conrast, when the cells were treated with higher doses of TanIIA for longer treatment periods, autophagic cell death contributed to apoptosis. Furthermore, generation of ROS occurred in a dose-dependent manner and pretreatment with NAC, a selective ROS scavenger, blocked the coexistence of Beclin-1 autophagy and caspase-dependent apoptosis. In conclusion, our findings provide strong evidence that TanIIA may be a potential therapeutic drug against osteosarcoma. Moreover, its cytotoxity can be enhanced with ROS agonists.

[Treatment of type C3 distal radius fractures with AO 2.4 mm locking plate system after manipulative reduction].

OBJECTIVE: To summarize our experiences in the treatment of type C3 (AO/OTA) distal radius fractures fixed with AO 2.4 mm locking plates combined with percutaneous pinning after manipulative reduction. METHODS: From May 2009 to March 2012, 19 patients (2 cases of both sides) with type C3 (AO/OTA) distal radius fractures were treated with volar locking plates combined with percutaneous pinning for distal radius after manipulative reduction. Among the patients, the average age was (45.3 ± 17.4) years old (ranged, 31 to 66 years old). The fracture were complicated with ulnar styloid fracture in 14 wrists and 6 wrists had distal radioulnar joint instability. All the patients had closed fracture and the mean duration was (6.7 ± 3.5) days (4.5 to 9 days). The Henry approach was applied to expose the fracture site. Joint capsule and ligaments were retained for indirect reduction. After indirect reduction, the poking reduction technique was used to correct the residual compression, and congruence of distal ulnar radial joint was verified under fluorscopic guidance. Styloid process was first pinned percutaneously and then AO 2.4 mm volar locking plate was used to support rigid fixation. The fractures complicated with distal radioulnar joint instability and ulnar styloid fracture were treated with forearm plaster support in supination for 6 weeks. RESULTS: Nineteen patients (21 wrists) were followed up for an average duration of 10.5 months (ranged, 7 to 17 months). Radiographic bone union of distal radius was achieved in all cases, nonunion of the ulnar styloid occurred in 3 cases, and no distal radioulnar joint instability occurred. Tendon irritation was found in 2 cases and disappeared after the internal fixation was removed. The volar tilt, radial angle, radial length, incongruence of articular surface and distal radioulnar joint were observed at the follow-up. According to Batra and Gupta scoring system, 13 wrists were assessed to have a score of more than 80, 5 wrists 70 to 90, 3 wrists less than 70. Meanwhile, the subjective and objective evaluation was executed,range of motion of wrist, residual deformity and complications were observed. According to Sarmiento's modification of the system of Gartland and Werley, 17 wrists got an excellent result, 3 good and 1 fair. CONCLUSION: Type C3 (AO/OTA) distal radius fractures could be managed with manipulative reduction. Locking plate internal fixation combined with percutaneous pinning can offer enough support for early mobilization and rehabilitation, resulting in a better clinical outcome and satisfactory prognosis.