Expression of Fibroblast Growth Factor 4 in a Rat Model of Polydactyly of the Thumb Induced by Cytarabine.

BACKGROUND The aim of this study was to assess the expression and mechanisms of fibroblast growth factor 4 in polydactyly of the thumb induced by cytarabine. MATERIAL AND METHODS Rats were intraperitoneally injected with cytarabine at different gestation periods (12.5 days, 13.5 days, and 14.5 days) to establish a polydactyly of the thumb model. Then, the expression of FGF4 in polydactyly was studied by whole-mount in situ hybridization. We used hematoxylin & eosin stain and cartilage stain to investigate the development of the skeleton and tissues in the embryo. Pictures were taken to determine the general shape of the deformity, then X-rays were taken to detect bone distortion of the rats born with a congenital malformation. RESULTS In the experimental group (11.5 days, 12.5 days, 13.5 days, and 14.5 days), whole-mount in situ hybridization showed that the FGF4 expression at the tip of the embryonic limb bud was significantly increased compared with the control group and FGF4 was distributed in a wider range and lasted longer than in the control group (P<0.01). HE staining and cartilage staining showed that there was an extra metacarpal bone and a phalanx in the rats with polydactyly of the thumb (P<0.01). Images of the deformed limbs showed polydactyly and syndactyly of the thumb in the rats. Further X-ray examination revealed 1 extra metacarpal bone and 1 extra phalanx. CONCLUSIONS Cytarabine can induce polydactyly and syndactyly of the thumb in rats. In this process, cytarabine can induce the expression of FGF4 on the tip of the embryonic limb bud, which further leads to abnormal development of the embryonic limb bud and eventually causes a congenital deformity.

Biosynthesis and characterization of zinc oxide nanoparticles from Artemisia annua and investigate their effect on proliferation, osteogenic differentiation and mineralization in human osteoblast-like MG-63 Cells.

The Biocompatibility and stability of nanoparticles using plants have been widely investigated due to its applications in the biomedical industry. Currently, there is a growing interest in nanoparticles in bone remodelling. Artemisia annua is an herbal plant commonly used in the treatment of various ailments. This study investigated the zinc oxide nanoparticles (ZnO NPs) using the green synthesis technique from A. annua and the effects of A. annua ZnO-NPs on osteoblast differentiation and inhibition of osteoclast formation. The formulated ZnO-NPs from A. annua were characterized by using various spectroscopic and microscopic methods Fourier transform-infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD), and UV-Visible spectroscopy. The disc diffusion method was adopted to test the antimicrobial efficacy of ZnO-NPs. The viability of MG-63 cells were assayed by MTT test and Osteogenic-related assays like Real-time PCR and Mineralization assay were adopted to determine the effects of A. annua ZnO-NPs on the multiplication and differentiation of human osteoblast-like MG-63 cells. The characterization of A. annua ZnO-NPs revealed the crystalline nature with high zinc content and the presence of bioactive compounds from A. annua extract. The synthesized A. annua ZnO-NPs indicate significant antimicrobial potential. Besides, A. annua ZnO-NPs enhanced the proliferation, differentiation, and mineralization without causing significant cytotoxic impact on MG-63 cells. These effects indicate that A. annua ZnO-NPs can both stimulate bone formation via the differentiation of MG-63 cells. Hence, it was concluded that A. annua ZnO-NPs can be a promising agent for the treatment of bone deformities and bone-related diseases, however further research also required to explore the clear mechanism of A. annua ZnO-NPs in the formation and differentiation of MG-63 cells.

Salvianolic acid B (Sal B) alleviates the decreased activity induced by prednisolone acetate on osteoblasts by up-regulation of bone formation and differentiation genes.

Glucocorticoids (GCs) are widely used to treat a variety of autoimmune diseases, but long-term use can lead to osteoporosis. To elucidate the mechanism of osteoporosis caused by glucocorticoids and to find effective protective drugs/foods, osteoblasts treated by prednisolone acetate were studied and salvianolic acid B (Sal B) was added to osteoblasts. The results showed that Sal B increased the activity of ALP and stimulated the expression of ALP that had been suppressed by prednisolone acetate. To further study the mechanisms of the protective effect of Sal B on osteoblasts treated with prednisolone acetate, the effects of gene expression involved with bone formation and differentiation were studied. The results show that the mRNA and protein expression of Runx2, Osx, OCN, IGF-I, Col-I and HO-I was up-regulated by Sal B. In conclusion, by stimulating the osteoblast activity and the expression of genes related to bone formation and differentiation, Sal B had a protective effect on osteoblasts that had been treated with prednisolone acetate.

Anti-Inflammatory Effects of Licochalcone A on IL-1ß-Stimulated Human Osteoarthritis Chondrocytes.

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), has been reported to have anti-inflammatory activity. In this study, we evaluated the anti-inflammatory effects of Lico A on IL-1ß-stimulated human osteoarthritis chondrocytes and investigated the possible mechanism. Results demonstrated that Lico A treatment significantly inhibited PGE2 and NO production induced by IL-1ß. IL-1ß-induced iNOS and COX-2 expression were also inhibited by Lico A. Lico A inhibited MMP1, MMP3, and MMP13 production in IL-1ß-stimulated chondrocytes. Lico A also inhibited IL-1ß-induced phosphorylation of NF-κB p65 and IκBα. Meanwhile, Lico A was found to upregulate the expression of Nrf2 and HO-1. However, Nrf2 siRNA reversed the anti-inflammatory effects of Lico A. In conclusion, our results suggested that Lico A showed anti-inflammatory effects in IL-1ß-stimulated chondrocytes by activating Nrf2 signaling pathway.

Thymoquinone Inhibits IL-1ß-Induced Inflammation in Human Osteoarthritis Chondrocytes by Suppressing NF-κB and MAPKs Signaling Pathway.

Thymoquinone, an active ingredient isolated from Nigella sativa, has been reported to have anti-inflammatory effects. However, the anti-inflammatory effect of thymoquinone on IL-1ß-stimulated osteoarthritis chondrocytes remains unclear. In this study, we designed to investigate the anti-inflammatory effects and elucidated the underlying mechanism of thymoquinone on IL-1ß-stimulated human osteoarthritis chondrocytes. The effects of thymoquinone on inflammatory mediators COX-2, iNOS, NO, PGE2, as well as MMP-1, MMP3, MMP13 production were detected. The results demonstrated that thymoquinone concentration-dependently inhibited IL-1ß-induced COX-2, iNOS, NO, and PGE2 production. Thymoquinone also suppressed IL-1ß-induced MMP-1, MMP3, and MMP13 production. We found that thymoquinone significantly inhibited IL-1ß-induced NF-κB activation and IκBα degradation. In addition, thymoquinone was found to suppress IL-1ß-induced mitogen-activated protein kinases (MAPKs) activation. In conclusion, thymoquinone inhibited IL-1ß-induced inflammatory mediator production by inhibition of NF-κB and MAPKs signaling pathways in osteoarthritis chondrocytes. Thymoquinone may be a potential agent in the treatment of osteoarthritis.

Developmental abnormalities in rat embryos leading to tibial ray deficiencies induced by busulfan.

BACKGROUND: Little is known about the developmental changes associated with tibial ray deficiencies. The aim of this study was to detect cell death, proliferation, and gene expression that result in tibial ray deficiencies. METHODS: We induced tibial ray deficiencies in rat embryos using a teratogenic agent (busulfan) and observed the developmental changes in 1126 hindlimbs. We performed Nile blue staining, whole mount in situ hybridization for fibroblast growth factor 8 (Fgf8), bone morphogenetic protein 4 (Bmp4) and Sonic hedgehog (Shh), terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) and assessment of cell proliferation by 5-bromo-2'-deoxy-uridine (BrdU)/anti-BrdU immunohistochemistry. RESULTS: In situ hybridization showed reductions in Fgf8 and Bmp4 expression. Histological examination showed a delay of mesenchymal condensation, increased mesenchymal cell death, decreased mesenchymal cell proliferation, and a reduction in the number of mesenchymal cells. These abnormalities may cause hypoplasia of the limb. Bmp4 expression was markedly reduced in the anterior mesenchyme. Shh was expressed in the posterior mesenchyme. We suggest that the posterior skeletal elements may be fully formed owing to Shh expression, but the anterior skeletal elements may be underdeveloped owing to an intense reduction of Bmp4 expression in the anterior mesenchyme, causing hypoplasia of the tibial ray. CONCLUSIONS: The combined effects of increased cell death, decreased cell proliferation, reduction of Fgf8 expression, and intense reduction of Bmp4 expression in the anterior mesenchyme may play an important role in the development of tibial ray deficiency induced by busulfan.

[Reform of the pedicled abdominal flap and its clinical application].

OBJECTIVE: To investigate the closing method of wound after removal of the traditional pedicled abdominal flap. METHODS: According to the design, the pedicled abdominal flaps were cut and lifted, and then the incision were extended from both sides on base of the flap to anterior superior iliac spine, respectively. After separating on superficial fascia, two flaps were obtained. The wound of donor site was closed completely by these two pedicled flaps. Twelve patients with skin defects on hands or forearms were treated using the reformed method of traditional pedicled abdominal flap. RESULTS: All of the 12 reformed pedicled abdominal flaps survived, and only one had local necrosis on the distal part of the abdominal flap, about 1.5 cm x 2.0 cm. CONCLUSION: This new design could provide a good method to close the abdominal wound after removal of pedicled abdominal flap.

Developmental failure of phalanges in the absence of growth/differentiation factor 5.

Growth/differentiation factor 5 (GDF5) is a member of the bone morphogenetic protein (BMP) family, which has been implicated in several skeletogenic events including cartilage and bone formation. To study the role of GDF5, we analyzed digit development in brachypodism (bp) mice, which carry functional null mutations of the Gdf5 gene and exhibit a reduction in the length of digit bones and loss of the middle phalanges. In situ detection of apoptosis and whole-mount detection of cell death showed abnormal apoptosis in the developing phalanges of bp mice. In situ hybridization in bp mice showed overexpression of Gdf5 mRNA in the developing phalanges where apoptotic cells were increased. In addition, bp mice exhibited excessive apoptosis in the interdigital regions. The condensed mesenchymal cells were progressively decreased in the developing phalanges and failed to form cartilage models of the middle phalanges. These findings show that excessive apoptosis in the absence of GDF5 results in developmental failure of the phalanges. We conclude that GDF5 is essential for maintenance and growth of the developing phalanges.

[The effect of liposome-carried metallothionein on secondary venous ischemia-reperfusion injury in a rat flap].

OBJECTIVE: To investigate the effect of liposome-carried metallothionein (lipo-MT) on secondary ischemia-reperfusion injury in the rat island flap. METHODS: An abdominal island flap was created in the Wistar rat. The animals were divided into four groups: the sham group, the secondary ischemia-reperfusion group, the group treated with blank liposome and the group treated with lipo-MT. The malondialdehyde (MDA) content, the myeloperoxidase (MPO) activity was assayed immediately, at 30 minutes and 7 days after the secondary venous ischemia-reperfusion. The level of endothelin (ET) and lactic dehydrogenase (LDH) of the rat plasma was measured at 30 minutes after secondary venous ischemia-reperfusion. The content of MT of the flap was assayed by Cd-hemoglobin saturation method at 7 days after the operation. RESULTS: The treatment of lipo-MT significantly decreased the content of MDA, MPO of the flap, decreased the activity of ET, LDH of the rat palsma, increased the content of MT of the flap and improved the flap viability. CONCLUSION: Lipo-MT can improve flap survival by reducing ischemia-reperfusion injury.

[Transferring neurovascular rectus femoris muscle segment for treatment of facial paralysis].

OBJECTIVE: To investigate a new technique for functional treatment of chronic facial paralysis. METHODS: Based on anatomy of intramuscular neurovascular structure in the rectus femoris muscle, 7 consecutive patients with facial paralysis were treated by using a technique of microsurgically free-transferring neurovascular rectus femoris muscle segment to the face in one-stage. Follow-ups were 10 to 24 months. RESULTS: All of the 7 patients showed significantly improvement in the appearance of the oral commissure and oral competence. No complications occurred in the donor site. CONCLUSIONS: The above mentioned technique may have the advantages of preventing the intramuscular nerve and vessel from the surgical injury during splitting the muscle. It could also maintain the transferred muscular segment in a proper tension in the recipient site.

[Effect of superoxide dismutase on adhesion molecules expression in skeletal muscle ischemia/reperfusion injury in rats].

OBJECTIVE: To study the effect of SOD on the expression of CD11b/CD18 and ICAM-1 after skeletal muscle ischemia / reperfusion injury in rats. METHODS: Establish the rat model of hind limb ischemia / reperfusion. One hundred and two rats were divided into 5 groups: sham operation group (Group I n = 6), ischemia group (GroupII n = 6), ischemia / reperfusion injury group (Group III n = 30), normal saline treatment group (Group IV n = 30), superoxide dismutase treatment group (Group V n = 30). Flow cytometric analysis and immunohistochemistry were used to assess the expression of CD11b/CD18 on the leukocytes, ICAM-1 in skeletal muscle at the end of ischemia and 1h, 2h, 4h,8h,12h after reperfusion. The changes of MDA in the plasma,MPO in the skeletal muscle and the tissue morphology were studied too. RESULTS: In group III,the expression of CD11b/CD18, ICAM-1,MDA,MPO were increased evidently. The longer time of reperfusion,the more serious the skeletal muscle's injury was. In group V, the changes were ameliorated as compared with group III (P < 0.05), group IV has no difference with group III. CONCLUSION: These data indicate that the expression of CD11b/CD18 and ICAM-1 were significantly correlated with skeletal muscle ischemia/reperfusion injury. SOD can inhibit the expression of free radicals and the adhesion molecules,consequently skeletal muscle ischemia/reperfusion injury is prevented.