Objective:To explore the clinical effects of a single stage reconstruction and revascularization using a free Flow-through chimeric anterolateral thigh perforator (ALTP) flap in Gustillo III C limb injuries.Methods:From January, 2010 to December, 2017, 17 patients with Gustillo III C injury of extremities were repaired with Flow-through chimeric ALTP in emergency surgeries. The patients were 16 males and 1 female with mean age of 32.4 (19-55) years. The size of wounds ranged from 16 cm×8 cm-45 cm×30 cm. The injured arteries were Flow-through anastomosed with the descending branch of the lateral circumflex femoral artery to regain blood flow. The deep dead space was filled with vastus lateralis flap, and the skin flap and fascia flap were used for superficial covering. The donor site was closed directly in 6 patients, simultaneous skin grafts were applied in 11 patients.Results:The followed-up time ranged 5 to 60 (average 21.8) months, and conducted by clinic visits and by telephone or WeChat interviews. Fifteen flaps survived, and 2 flaps failed with limb amputated. Six patients were repaired with skin and fascia flaps and 11 patients received flaps comprising the skin, fascia and vastus lateralis muscle. Four flaps healed in stage I. Partial necrosis were observed in 11 flaps. Of which, 1 healed by change of dressing and 10 healed by skin grafting. No complications occurred in donor site in all patients.Conclusion:The Flow-through chimeric ALTP flap can be used in stage I reconstruction of the blood supply of limbs and meanwhile achieve the 3-D repair of defects. It is one of the reliable methods in the reconstruction of Gustillo III C injuries in the extremities.

Objective:To study the clinical effect and hemorheological changes of hyperbaric oxygen (HBO) combined with Tuina therapy on patients with cervical spondylosis of vertebral artery type (CSA).Methods:One hundred and ten patients with CSA, admitted to the Affiliated Hospital of Guilin Medical University from January 2018 to January 2019, were randomly divided into observation group and control group according to the random number table, with 55 cases in each group. The control group was treated with Tuina therapy of traditional Chinese medicine, while the observation group was treated with HBO based on the treatment of the control group. The clinical effect, hemorheological changes, cervical vertebral movements, the levels of neuropeptide Y (NPY) and urotensin Ⅱ(UⅡ) in plasma were tested and recorded before and after treatment.Results:After treatment, the total effective rate of the observation group (94.55%) was significantly higher than that of the control group (81.82%) ( P<0.05). The peak systolic blood flow velocity and the mean blood flow velocity of vertebrobasilar artery in the observation group after treatment were significantly increased as compared with the control group ( P<0.05). The angles to anterior, posterior, left, and right of cervical vertebral movement of the observation group after treatment were significantly larger than those of the control group ( P<0.05). The levels NPY and UⅡ in plasma of both groups after treatment were significantly lower than those before treatment ( P<0.01), and the levels NPY and UⅡ in the observation group were significantly lower than those in the control group after treatment ( P<0.05). Conclusion:HBO combined with Tuina therapy can improve the curative effect on CSA, by improving cervical vertebral movement and hemorheological characteristics, and lowering the levels of NPY and UⅡ in plasma in CSA patients.

Objective:To study the clinical effect and hemorheological changes of hyperbaric oxygen (HBO) combined with Tuina therapy on patients with cervical spondylosis of vertebral artery type (CSA).Methods:One hundred and ten patients with CSA, admitted to the Affiliated Hospital of Guilin Medical University from January 2018 to January 2019, were randomly divided into observation group and control group according to the random number table, with 55 cases in each group. The control group was treated with Tuina therapy of traditional Chinese medicine, while the observation group was treated with HBO based on the treatment of the control group. The clinical effect, hemorheological changes, cervical vertebral movements, the levels of neuropeptide Y (NPY) and urotensin Ⅱ(UⅡ) in plasma were tested and recorded before and after treatment.Results:After treatment, the total effective rate of the observation group (94.55%) was significantly higher than that of the control group (81.82%) ( P<0.05). The peak systolic blood flow velocity and the mean blood flow velocity of vertebrobasilar artery in the observation group after treatment were significantly increased as compared with the control group ( P<0.05). The angles to anterior, posterior, left, and right of cervical vertebral movement of the observation group after treatment were significantly larger than those of the control group ( P<0.05). The levels NPY and UⅡ in plasma of both groups after treatment were significantly lower than those before treatment ( P<0.01), and the levels NPY and UⅡ in the observation group were significantly lower than those in the control group after treatment ( P<0.05). Conclusion:HBO combined with Tuina therapy can improve the curative effect on CSA, by improving cervical vertebral movement and hemorheological characteristics, and lowering the levels of NPY and UⅡ in plasma in CSA patients.

Objective:To investigate whether celastrol can increase the cytotoxic activity of γδ T cells against osteosarcoma (OS) cells line HOS through TRAIL way. Methods:The death receptors 4/5 (DR4/5) protein levels in the OS cell lines HOS was in-vestigated by Western blot analysis. Zoledronate ( ZOL) was used to induceγδT cells from PBMCs of healthy volunteers.γδT cell cy-totoxicity against HOS cells was investigated by a standard lactate dehydrogenase release assay ( LDH ) . Results:Celastrol increased DR4/5 protein expression in HOS ( P<0. 05 ) .γδ T cells from PBMCs of healthy volunteers showed cytotoxicity against HOS ( P<0. 05). Following co-culture with HOS pre-treated with celastrol (1 μmol/L) for 24 h,γδ T cells showed significantly higher cytotoxicity against HOS (P<0. 05). The induction of DR4/5 molecules increased lysis of HOS by γδ T cells which was abolished by addition of a blocking TRAIL antibody. Conclusion:Celastrol can enhance γδ T cells'cytotoxic activity against OS cells line HOS through TRAIL way.

BACKGROUND:There are less studies on static magnetic field, and its biological effects are stil unclear. OBJECTIVE:To investigate the static magnetic field effect on the secretion of nerve growth factor from Schwann cel s. METHODS:Schwann cel s were randomly assigned into three groups:0.05 mT sciatic magnetic field group, 0.1 mT sciatic magnetic field group and control group. At 2 days of inoculation, the cel s were exposed to the sciatic magnetic field, 12 hours per day. Cel s in the control group were not exposed to the sciatic magnetic field. After 6 days of exposure, RT-PCR method was used to detect nerve growth factor mRNA expression in Schwann cel s, and ELISA was adopted to detect the level of nerve growth factor secreted from Schwann cel s. RESULTS AND CONCLUSION:The mRNA expression and secreted level of nerve growth factors in the culture supernatant were significantly higher in the two sciatic magnetic field groups than the control group (P<0.05), especial y in the 0.1 mT group. However, there was no difference between the 0.05 mT and 0.1 mT groups (P>0.05). These findings indicate that the static magnetic field with certain intensity can promote the secretion of nerve growth factors from Schwann cel s.

Objective To investigate the cytotoxic activities of γδT cells against methotrexate (MTX)-resistant osteosarcoma (OS) cells.Methods The MTX-resistant U2OS cell line (U2OS/MTX) was established by in vitro exposing the parental cells to MTX at stepwise-increasing concentrations.Periph-eral blood mononuclear cells ( PBMCs) were isolated from healthy subjects and stimulated with zoledronate ( ZOL) in combination with IL-2 to induce the proliferation ofγδT cells.The cytotoxicity ofγδT cells against U2OS/MTX cells was analyzed by using a standard lactate dehydrogenase release assay (LDH).Flow cy-tometry analysis and ELISA were performed to detect the expression of CD69 and IFN-γby γδT cells, re-spectively.Results The γδT cells derived from healthy subjects showed cytotoxicity against the U2OS/MTX cells.Moreover, stronger cytotoxic activities of γδT cells were detected after pretreatment of U2OS/MTX cells with ZOL (1 μmol/L) for 24 hours (P<0.01).Stimulation with U2OS or U2OS/MTX cells in-creased the expression of CD69 onγδT cells and enhanced the secretion of IFN-γbyγδT cells (P<0.05). Increased expression of CD69 and enhanced secretion of IFN-γwere induced in γδT cells in response to stimulation with ZOL-pretreated U2OS/MTX cells (P<0.01).Conclusion TheγδT cells showed cytotox-icity against U2OS/MTX cells.Pretreatment of U2OS/MTX cells with ZOL could enhance the cytotoxic ac-tivity of γδT cells.

Objective To investigate the effects of metal particles produced by metal prosthesis on oxidative stress levels in human mononuclear cells . Methods Mononuclear cells obtained from peripheral blood of 15 healthy volunteers .Coculture mononu-clear cells with iron particles , cobalt particles , chromium particles , titanium particles and physiological saline , respectively .Levels of reactive oxygen species ( ROS) , malondialdehyde ( MDA) , superoxide dismutase ( SOD) and glutathione peroxidase ( GSH) were as-sessed in mononuclear cells at 6 h, 12 h, 24 h, 36 h, 48 h, respectively . Results Levels of ROS and MDA were higher in the parti-cle groups than that in the control group at 6 h, 12 h, 24 h, 36 h, 48 h, respectively (P<0.05).Levels of ROS and MDA were higher in cobalt particle, chromium particle and iron particle groups than that in the titanium particle group (P<0.05).Levels of SOD and GSH were lower in particle groups than that in the control group (P<0.05).Levels of SOD and GSH were lower in cobalt particle , chromium particle and iron particle groups than that in the titanium particle group (P<0.05). Conclusion Oxidative stress levels increased and anti-oxidant levels decreased in mononuclear cells when cocultured with metal debris .Cobalt particles , chromium parti-cles and iron particles induced higher changes than that in titanium particles .Oxidative stress may play an important role in metal deb-ris-mediated osteolysis .

Objective To discuss parallel fibular osteoseptocutaneous flap apply in heel defect reconstruction Methods From February,2006 to December,2011,parallel fibular osteoseptocutaneous flap were used to repair calcaneus and skin defects in 4 cases.The causes included road accident in 2 cases and strangulation in 2 cases.The defect locations were at the back of the calcaneus (1/2 of calcaneus in 2 cases and 2/3 in 2 cases,respectively).Results Followed up 24-72 months,all cases achieved bone union in allograft and had no complications of absorption,infection and repulsion.The flaps survived completely in 4 cases ; the distal flaps necrosed partly in 1 case and healed by dressing.According to USA foot-malleolus scores of AOFAS,1 case was excellent,2 good,and 1 fair.Conclusion Parallel fibular osteoseptocutaneous flap reconstruction heel defect can recover patients visage and heel function,improve their life quality.

Objective:To investigate the effect of typeⅠIFN on the cytotoxic activity ofγδT cells from peripheral blood mono-nuclear cells ( PBMCs) of healthy donors and osteosarcoma patients stimulated by zoledronate ( Zol) and IL-2 against OS.Methods:PBMCs from healthy donors and osteosarcoma patients were stimulated with Zol+IL-2 or Zol+IL-2+typeⅠIFN,respectively.After 14 days of culture,ex vivo expandedγδT cells were assessed by flow cytometry.γδT cell cytotoxicity against target cells was analyzed using a standard lactate dehydrogenase release assay.IFN-γsecreted fromγδT cells was determined by ELISA kits.Results:γδT cells from PBMCs of healthy donors and patients with OS were selectively expanded by Zol+IL-2 or Zol+IL-2+typeⅠIFN in vitro,respectively, and showed cytotoxicity against OS.In addition,γδT cells pre-treated by TypeⅠIFN showed significantly higher cytotoxicity against OS cells.Conclusion:Type I IFN can enhance humanγδT cells’ cytotoxic activity against OS.

OBJECTIVETo investigate the cytotoxic effect of γδ T cells from osteosarcoma patients against interferon-γ (IFN-γ)-treated osteosarcoma cells in vitro.

METHODSHuman γδ T cells were amplified by zoledronate from peripheral blood cells of osteosarcoma patients. The expression of Fas on the osteosarcoma cells were measured by flow cytometry and quantitative real-time PCR analysis before and after IFN-γ treatment. The cytotoxicity of γδ T cells against osteosarcoma cells was evaluated with LDH assay.

RESULTSIFN-γ significantly enhanced the susceptibility of the osteosarcoma cell lines HOS and U2OS to the cytotoxicity of γDelta; T cells from osteosarcoma patients (P<0.01). IFN-γ obviously up-regulated the expression of Fas in HOS and U2OS cells (P<0.01). Anti-FasL mAb failed to inhibit the cytotoxicity of γδ T cells in untreated osteosarcoma targets (P>0.05), but significantly impaired γδ T cell cytotoxicity in IFN-γ pre-treated osteosarcoma targets (P<0.01).

CONCLUSIONIFN-γ can enhance the cytotoxic effect of human γδ T cells from osteosarcoma patients against osteosarcoma cells in vitro.

Bone Neoplasms ; metabolism ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; Humans ; Interferon-gamma ; pharmacology ; Osteosarcoma ; immunology ; metabolism ; Receptors, Antigen, T-Cell, gamma-delta ; immunology ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects ; immunology ; fas Receptor ; metabolism