To investigate the pathological changes in gunshot wound track in limbs of dogs produced under a hot and humid environment, 8 dogs were randomly divided into two groups: hot and humid environment group (HHE) and normal environment group(NE). Gross examination, optical microscopic examination (OM), electron microscopic examination (EM) were performed, and ATP, ADP, AMP content, energy charge (EC) in the gunshot wounds were observed at 4, 8, 12, 24h after wounding. The results indicated that 6～8 h after the injury, the wound tracks and discolored area of tissue in HHE group were more distinguished and enlarged than those in NE group, and the swelling of wounded limbs was obvious. Some of the wounds began to be odoriferous. The signs of infection began to be obvious. While the wounds of NE group did not give off obvious odor of decay until 12～24h after the injury. Under OM and EM, necrosis and damage of muscle fibers in the wound tracts were more obvious in the HHE group than that in NE group, showing a tendency of deterioration. In contrast the changes in the injured tissues in NE group showed a tendency of improving. ATP content of the injured tissues in the contusion region of HHE group was lower than that of NE group at 8h, ATP content of tissues in the shock region of HHE group was decreased continuously and was lower than that of NE group at 12h. ADP content of tissues in the contusion area of HHE group was lower than that of NE group at 8h and also in shock area at 12h. AMP content of tissues in the contusion area of HHE group was higher than that of NE group at 12h, whereas EC of tissues in the contusion area of HHE group was lower than that of NE group at 8h and 12h. Histopathological changes in tissues in the gunshot wound were serious and deteriorated gradually in hot and humid environment. At the same time the energy storage of muscle tissue in the gunshot wound track was lower and decreased more remarkably in this environment. Therefore early and complete debridement of gunshot wounds inflicted in hot and humid environment should be emphasized.

Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.

Objective To study the molecular mechanism of glutamate receptor-6 (GluR6) in the pathogenesis of epilepsy. Methods Seizure model of SD rats was induced by intraperitoneal injection of kainate (KA). Immunoprecipitation and immunoblotting were performed to examine the interactions of GluR6 and MLK3 with PSD95 at various time points after KA injection. The effect of Tat-GluR6-9c on the MLK3 phospharylation induced by kainate was observed with immunoblotting and immunohistochemistry. Results The assembly of GluR6 and MLK3 with PSD95 was induced after KA hippocampal CA3 region, and bagan to decrease one day later. Pretreatment after KA injection in CA3 region (P<0.05). Conclusion KA induces the assembly of the GluR6-PSD95-MLK3 signaling module and subsequently activates MLK3, which ultimately results in brain injury.

AIM: To explore the mutual effect of co-culture of wild-type astrocytes (ASC) and motor neurons VSC4.1 (VSC) on the respective ability to produce reactive oxygen species (ROS). METHODS: The inhibition rates of cell growth in ASC and VSC in co-culture or independent culture was detected after exposed to excitatory stimulus by MTT method. Real-time observation of ROS production by ASC and VSC labeled with Hoechst 33342 was detected by confocal microscopy under the conditions of co-culture or independent culture. RESULTS: Higher concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in ASC alone, while lower concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in VSC only. Real-time observation by confocal microscopy showed that ROS production by VSC under the condition of co-culture, which showed a notable increase at 15 min, was significant less than that in independent culture, which peaked at 5 min and was gradually decreased. ROS production by ASC in co-culture began to increase significantly at 10 min. CONCLUSION: Compared to independent culture, ASC reduces the resting ROS production by co-cultured with VSC, while ASC prolongs the duration of ROS production by VSC after exposed to excitatory stimulus.

Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice.Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma.When the volume of implanted tumor reached 100-150 mm3,murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18,IL-18 gene-expressing adenovirus named Ad-IL-18,phosphate buffer saline(PBS),respectively.The tumor size was measured at an interval of 4 days for 9 weeks.Hematoxylin and eosin (HE)staining was performed to observe the morphological changes of tumor cells.The protein expression of IL-18 and E1A.microvessel density in tumor tissue,and apoptosis of tumor xenografts were detected by immuno fluorescence assay,immunohistochemistry and in situ end labeling technique (TUNEL).respectively.Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor.Forty-four days after the treatment,the mean tumor volume was 1039.378±29.67 mm3 in ZD55-IL-18-treated mice.significantly smaller than that in Ad-IL-18 treated mice(2900.46±62.65 mm3)and PBS-treated mice(3980.24±63.78 mm3).HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice.Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs.AD55-IL-18-treated mice(83.4%±3.2%vs 24.4%±2.1%.P＜0.01).Moreover,immunofluorescence assay revealed the presence of E1A protein in tumor tissue.A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice(P＜0.01).The apoptosis rate in tumor cells from high to low was 86.28%±3.25%in ZD55-IL-18-treated mice,43.67%±3.46%in Ad-IL-18-treated mice,and 10.73%±2.48%in PBS-treated mice;there was a significant difference between the three groups(all P＜0.05).Conclusion The oncolytic adenovirus expressing human IL-18 gene,ZD55-IL-18,has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.

Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Objective:to investigate the clinical feature and dynamic changes of the cervical dural sac and spinal cord during neck flexion in Hirayama disease(juvenile muscular atrophy of distal upper extremity).Methods:Clinical data were taken and MRI in neutral neck position and a fully flexed neck position were performed on 27 cases of Hirayama disease.Results:(1)All patients were consistent with the diagnostic criteria of Hirayama disease who had asymmetric muscular atrophy and weakness of the hand and forearm.All patients were young males and right handed of whom 77.8% had initial symptoms before they were 19 years old.More patients(20 cases,74%)had muscular atrophy in the right hand than in the left at onset.The duration after disease onset was from 2-72 months[(26.48?15.57)months].(2)In neutral neck position by MIR examination,16 patients showed abnormal cervical curvature,14 showed atrophy of the lower cervical cord and 2 patients had intramedullary abnormal high signal.(3)In a fully flexed position of the neck,all patients showed forward displacement and flattening of the lower cervical cord,and a crescent-shaped high signal area behind the cord.(4)The crescent-shaped area was enhanced on T1-weighed imaging and disappeared after the patient returned to a neural position in one case.Conclusion:Hirayama disease occurs mainly in young males.There are obviously dynamic changes of the cervical cord during neck flexion in Hirayama disease by MRI examination,which can help the doctor make diagnosis in the early stage.

Objective To investigate the effect of acetyl-L-carnitine (ALC) preconditioning on the PC12 cell apoptosis induced by oxygen-glucose deprivation.Methods PC12 cells were seeded in 96-well plates and randomly divided into 5 groups ( n =6 each):control group (group C),cell injury group (group Ⅰ) and preconditioning with different concentrations of ALC groups (groups A1-3 ).In group C,the cells were incubated with DMEM liquid culture medium containing glucose 0.5 g/L for 3 h.In groups Ⅰ and A1-3 the cells were incubated with DMEM liquid culture medium containing sodium hydrosulfite (Na2S2O4) 3 mmol/L and glucose 0.5 g/L for 3 h,and in addition the cells were pre-incubated with ALC 0.2,0.4 and 0.6 mmol/L for 24 h in groups A1-3 respectively.Cell viability was evaluated by MTF assay,while the apoptosis in cells was detected using TUNEL.The activities of ATPase and SOD and MDA content were also detected.Results Oxygen-glucose deprivation significantly increased the number of apoptotic cells and the content of MDA,and decreased the cell viability and activities of SOD and ATPase in group Ⅰ compared with group C ( P ＜ 0.05).Preconditioning with ALC significantly increased the cell viability and the activities of SOD and ATPaes,and decreased the number of apoptotic cells and the content of MDA in groups A1-3 compared with group Ⅰ ( P ＜ 0.05).Conclusion ALC preconditioning can attenuate PC12 cell injury induced by oxygen-glucose deprivation through inhibition of apoptosis in cells.

OBJECTIVE: To study the methods and techniques of free flap transfer bridged by posterior tibial vascular flap in treating large soft tissue defects in low limbs without usable recipient blood vessels. METHODS: Based on morphological observation and measurement of arterial pressure and blood flow, an antegrade and a retrograde vascular bridge flaps were designed using the healthy posterior tibial vessels to serve as vascular pedicles to carry two free flaps for transplantation. RESULTS: Eight cases of patient with one or two large soft tissue defects in the leg region were treated by the method. All the bridge flaps and free flaps survived well, and the defects were repaired completely. CONCLUSIONS: The results showed that it is an ideal method for using the posterior tibial vessels from the healthy limb to form vascular pedicles in repairing large soft tissue defects in patients without a usable recipient blood vessel.

Objective: To observe morphological change and diversity of periodontium and alveolar bone after tooth transplantation into artificial tooth socket. Methods: 6 dogs were divided randomly into 2 groups: 2 dogs were used as the controls and 4 used for the experiment. In the control group 4 teeth were autotransplanted into the inherent sockets. In the experiment group 4 teeth were autotransplanted into the artificial sockets. The dogs were sacrificed at the 16th week after operation. The healing condition of periodontal tissue and the remodeling of alveolar bone were examined. Results: None of the transplanted teeth in both groups was loosen or dropped. Mircro-CT examination showed that cancellous bone and bone trabecula around the transplanted teeth lined tightly,no significant difference of bone trabecula thickness was observed between the 2 groups. Hard tissue slice examination revealed that parodontium of both groups grew and adhered to the teeth,and the quantity of new-born bone between the top of alveolar ridge and the neck of transplanted teeth was fundamentally the same in the 2 groups. Conclusion: Autotransplantation of teeth into the artificial socket is similar to that into inherent socket.