Serotonergically dependent antihyperalgesic and antiallodynic effects of isoliquiritin in a mouse model of neuropathic pain.

Chronic neuropathic pain poses a significant health problem worldwide, for which effective treatment is lacking. The current work aimed to investigate the potential analgesic effect of isoliquiritin, a flavonoid from Glycyrrhiza uralensis, against neuropathic pain and elucidate mechanisms. Male C57BL/6J mice were subjected to chronic constriction injury (CCI) by loose ligation of their sciatic nerves. Following CCI surgery, the neuropathic mice developed pain-like behaviors, as shown by thermal (heat) hyperalgesia in the Hargreaves test and tactile allodynia in the von Frey test. Repetitive treatment of CCI mice with isoliquiritin (p.o., twice per day for two weeks) ameliorated behavioral hyperalgesia to thermal (heat) stimuli and allodynia to tactile stimuli in a dose-dependent fashion (5, 15 and 45 mg/kg). The isoliquiritin-triggered analgesia seems serotonergically dependent, since its antihyperalgesic and antiallodynic actions were totally abolished by chemical depletion of spinal serotonin by p-chlorophenylalanine, whereas potentiated by 5-HTP (a precursor of 5-HT). Consistently, isoliquiritin-treated neuropathic mice showed escalated levels of spinal monoamines especially 5-HT, with depressed monoamine oxidase activity. Moreover, isoliquiritin-evoked antihyperalgesia and antiallodynia were preferentially counteracted by the 5-HT1A receptor antagonist WAY-100635 delivered systematically or spinally. Of notable benefit, isoliquiritin was able to correct co-morbid behavioral symptoms of depression and anxiety evoked by neuropathic pain. Collectively, these findings demonstrate, for the first time, the therapeutic efficacy of isoliquiritin on neuropathic hypersensitivity, and this effect is dependent on the spinal serotonergic system and 5-HT1A receptors.

Artesunate Decreases ß-Catenin Expression, Cell Proliferation and Apoptosis Resistance in the MG-63 Human Osteosarcoma Cell Line.

BACKGROUND: This study aims to determine the effects of artesunate on proliferation, apoptosis and ß-catenin expression in the human osteosarcoma cell line MG-63. METHODS: MG-63 cells in the logarithmic growth phase were collected and cultured with different concentrations of artesunate (12.5 µg/mL, 25 µg/mL and 50 µg/mL) for 24 h, 48 h and 72 h. The total number of MG-63 cells and the morphological changes were observed under an inverted microscope. The MTT assay was adopted to test the inhibition rate (IR) of cell growth. The apoptosis rate was detected using annexin V/propidium iodide (PI) staining. Cell cycle distribution was identified by flow cytometry (FCM), and the expression levels of ß-catenin, cyclins and cyclin dependent kinases (CDKs) were measured using Western blotting. RESULTS: The results of the MTT assay indicated that artesunate could remarkably inhibit MG-63 cell proliferation compared with the rates in the untreated control group (0 µg/mL artesunate), and the inhibitory effect was dose-dependent. The apoptosis rate of MG-63 cells was elevated as the concentration of artesunate increased, and all the rates were significantly higher than that in the control group. Additionally, as the artesunate concentration increased, the proportion of MG-63 cells in G0/G1 phase gradually declined whereas that of cells in the G2/M and S phases increased. Western blotting confirmed that a higher concentration of artesunate reduced the expression levels of ß-catenin, cyclin A, cyclin D1 and CDK1 and increased the expression levels of cyclin B1; however, artesunate had no impact on CDK2 expression in MG-63 cells. CONCLUSION: These results demonstrated that artesunate can inhibit ß-catenin expression and cell proliferation as well as promote cell apoptosis in MG-63 cells, which indicates that artesunate may serve as a promising drug in the clinical treatment of osteosarcoma.

GIT1 gene deletion delays chondrocyte differentiation and healing of tibial plateau fracture through suppressing proliferation and apoptosis of chondrocyte.

BACKGROUND: Although tibial plateau fracture is an uncommon injury, its regulation is challenging and there are some influencing factors, including the effects of severe bone displacement, depression and cancellous bone cartilage, and inevitable cartilage damage. And GIT1 plays an important role in bone mass and 78 osteoblast cell migration. METHODS: The study used 72 C57/BL6 mice. A tibial plateau fracture model was established by using mice with the same number of GIT1 gene deletions (the experimental group) and their wild-type littermates (the control group). Joint and bone callus recovery were evaluated by X-ray and CT thin layer scans. Micro CT assay and histomorphometry were conducted in order to evaluate the volume of newly formed blood vessels. Type II collagen expression in tibial tissues after tibial plateau fracture were detected by immunohistochemistry after 7, 14 and 21 days. The number of proliferating cell nuclear antigen (PCNA) positive cells after tibial plateau fracture was tested by immunohistochemistry after 14 and 21 days. The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was conducted after 14 and 21 days in order to test chondrocyte apoptosis in tibial tissues after tibial plateau fracture. RESULTS: The GIT1 gene deletion group mice spent less time on the rotating rod than the control group mice (P < 0.05). Compared with the control group, postoperative recovery was retarded, because GIT1 gene deletion slowed down neovascularization after tibial plateau fracture (P < 0.05). Compared with the control group, mouse type II collagen expression significantly decreased in the GIT1 gene deletion group, and the proportion of PCNA positive cells significantly decreased (P < 0.05). The TUNEL results indicate that GIT1 gene deletion led to reduced chondrocyte apoptosis. CONCLUSION: GIT1 gene deletion can inhibit chondrocyte proliferation and apoptosis during the recovery of tibial plateau fracture, so as to delay chondrocyte differentiation and tibial plateau fracture healing.

Bridging external fixation versus non-bridging external fixation for unstable distal radius fractures: A systematic review and meta-analysis.

OBJECTIVE: A systematic review and meta-analysis was conducted to compare the relative effectiveness of bridging external fixation and non-bridging external fixation for distal radius fractures treatment. METHOD: Relevant literature were comprehensively searched using the PubMed, Springer Link, Karger Medical and Scientific Publishers, Chinese Biomedical Database (CBM) and Chinese National Knowledge Infrastructure (CNKI) databases without any language restrictions. STATA Version 12.0 software and Comprehensive Meta-analysis 2.0 were applied. RESULTS: A total of 905 patients with distal radius fracture from six eligible cohort studies were selected for statistical analysis. Our meta-analysis results indicate that the non-bridging cases had a higher risk of pin track infection, rupture of the extensor pollicis longus and nerve injury than the bridging cases. Subgroup analysis stratified by country indicated non-bridging patients showed evidence of an increased risk of pin track infection and higher risk of rupture of the extensor pollicis longus compared with the patients treated with bridging external fixation in the UK population. The follow-up results showed flexion degree of patients treated with non-bridging external fixation was slightly better than that of patients treated with bridging external fixation (P < 0.05). CONCLUSION: There is evidence in our systematic review and meta-analysis to support that bridging external fixation can reduce the incidence of pin tract infections and nerve injury compared to non-bridging external fixation, but have no significant difference in other complications and the recovery of wrist joint function. Bridging external fixation could therefore be a better choice in patients with distal radius fractures.

DACH1 Expresison in Osteosarcoma and Its Relationship with Proliferation and Angiogenesis.

The aim of this study was to investigate the expression of DACH1 in osteosarcoma as well as its relationship with cell proliferation and angiogenesis in the tumor. DACH1 expression was detected by immunohistochemical staining in the serial sections of the osteosarcoma. The microvessel density (MVD) was counted by CD34 immunohistochemical staining, and immunohistochemical staining of PCNA staining showed the cell proliferation. The impacts of DACH1 expression on tumor proliferation and angiogenesis were evaluated by statistics. The DACH1 had different expression patterns in different osteosarcoma. Conventional osteosarcoma showed stronger DACH1 staining (conventional vs. parosteal: P = 0.037; conventional vs. periosteal: P = 0.028) and more PCNA-positive tumor cells than parosteal and periosteal osteosarcoma (conventional vs. parosteal: P = 0.041; conventional vs. periosteal: P = 0.045), the difference was significant. In addition, conventional osteosarcoma showed more cytoplasmic staining of DACH1 than parosteal and periosteal (conventional vs. parosteal: P = 0.023; conventional vs. periosteal: P = 0.030). Parosteal and periosteal osteosarcoma showed no significant difference in DACH1 expression and cell proliferation index. On the other hand, DACH1 different expression patterns showed significantly different impacts on angiogenesis. In spite of the different subtypes of osteosarcoma, the MVD showed a significant difference in cytoplasmic and nuclear expression patterns of DACH1 (nuclear expression vs. cytoplasmic expression: 5.72 ± 1.19 vs. 9.65 ± 1.24, P = 0.042). Moreover, in the conventional osteosarcoma, the MVD also showed a significant difference in DACH1 cytoplasmic and nuclear staining (nuclear expression vs. cytoplasmic expression: 5.58 ± 0.71 vs. 13.65 ± 1.30, P = 0.019). However, the DACH1 expression intensity showed no significant different impacts on MVD of all kinds of osteosarcoma. DACH1 had different expression patterns and intensity. Cytoplasmic and nuclear expression of DACH1 might play different roles in cell proliferation and angiogenesis of osteosarcoma. Cytoplasmic DACH1 might promote cell proliferation and be associated with angiogenesis.

Changes of lipid peroxidation in carbon disulfide-treated rat nerve tissues and serum.

To further elucidate the mechanism and determine the biomarker of neuropathy induced by carbon disulfide (CS(2)), we performed a longitudinal observational study of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD) and total antioxidative capacity (T-AOC) in rat cerebral cortex, hippocampus, spinal cord and serum after 0, 2, 4, 8 and 12 weeks of CS(2) administration. CS(2) exposure was found to markedly increase ROS and MDA levels in cerebral cortex, hippocampus, spinal cord and serum of rats in both time and symptom-dependent manners. Although SOD activities slightly increased, there was a decrease in the GSH contents and GSH-Px, CAT activities in cerebral cortex, hippocampus, spinal cord and serum after 2, 4, 8 or 12 weeks' CS(2) intoxication and at gait score of 2, 3, or 4. The activities of T-AOC also decreased in all three nerve tissues and serum as time went on and symptom developed. Furthermore, significant correlations between LPO and gait abnormality were observed as symptom developed. Oxidation stress also resulted in Ca(2+) concentrations and calmodulin (CaM) levels increases in cerebral cortex, hippocampus and spinal cord. Thus, CS(2) intoxication was associated with elevation of lipid peroxidation (LPO) and reduction of antioxidant status, and the time and symptom-dependent changes of these indexes in rats' nerve tissues and serum suggested that ROS and concomitant LPO, at least in part, were involved in CS(2)-induced neuropathy.

Laparoscopic management of type I choledochal cyst in adults: cyst resection, assisted Roux-en-Y reconstruction and hepaticojejunostomy.

Due to improvement of instrumentation and surgeons' skills, the correction of congenital biliary tract anomalies has been performed by the laparoscopic approach. Because of the high rate of associated malignancy of the biliary system in middle-aged adults, treatment for choledochal cyst is necessary, especially in adult patients. We report on the laparoscopic excision and hepaticoduodenostomy of type I choledochal cysts in five adult patients. To facilitate the procedure, the creation of a Roux-en-Y reconstruction was performed with a minimal abdominal incision. All patients had an uneventful recovery with no major complications. Most were discharged on day 8 after the procedure. At a follow-up of two years, they were still asymptomatic, showing no cholangitis or anicteric. Laparoscopic management for choledochal cyst is an advantageous approach, so it is feasible and will probably become an accepted method in further clinical application.