BACKGROUND: The low survival rate of neuron cells is one of the main mechanisms of stem cell allograft, which might lead to the failure of allograft. Nuclear factor-KB (NF-kB) is one of main transcription factors for cell signaling transduction and participates in call proliferation and differentiation.OBJECTIVE: To study the differentiation of muscle-derived stem cells (MDSCs) into neuron-like cells induced by ciliary neurotrophic factor (CNTF) and Compound Salvia Miltiorrhiza Injection in vitro, and the expression of NF- kB.DESIGN, TIME AND SETTING: The experiment at cell molecular level was performed at the Oral Science Laboratory of the Second Affiliated Hospital of Liaoning Medical College from March to May 2008.MATERIALS: A total of 10 Sprague Dawley neonatal rats aged 7 days were supplied by Experimental Animal Center, Liaoning Medical University. CNTF (Sigma, USA) and Compound Salvia Miltiorrhiza Injection (Chiatai Qingchunbao Pharmacantical Co.,Ltd., China) were used in this study.METHODS: Rat MDSCs were harvested in vitro, pudfied by differential adherence and enzyme digestion, and incubated in 6-well plate. Samples in the induction group were incubated in DMEM containing CNTF for 24 hours. The medium was changed.Subsequently, samples were dnsed three times, and then incubated in serum-free DMEM supplemented with Compound Salvia Miltiorrhiza Injection for 5 hours. Samples in the control group were treated with serum-free DMEM.MAIN OUTCOME MEASURES: Neurofilament protein and NF-KB inhibitor protein expression were detected using reverse transcription-polymerase chain reaction (RT-PCR) and Westem blotting.RESULTS: No neurofilament protein expression was found in MDSCs before induction, and neurofilament protein-positive MDSCs were detected following induction. Results of gel electrophoresis and Westam blot showed that no significant differences in NF-kB inhibitor protein expression were determined in the control group, and NF-kB inhibitor protein expression was significantly decreased in the induction group after induction.CONCLUSION: CNTF and Compound Salvia Miltiorrhiza Injection could inhibit the activation of NF-kB and induce the differentiation of MDSCs into neuron-like cells.Zeng XY, Wang W, Sun L, Zhang L, Zeng LD.Differentiation of muscle-derived stem cells into neuron-like cells induced by ciliary neurotrophic factor and Compound Salvia Miltiorrhiza Injection in vitro.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu.2009;13(27): 5336-5340. [http://www.crter.cn http://en.zglckf.com]

Objective To discuss clinical outcomes of extraperitoneal pelvic packing (EPP) plus external fixation for hemodynamically unstable pelvic fractures. Methods We used EPP plus external fixation to control 15 cases of hemodynamically unstable pelvic fractures from further damage. They were 6 males and 9 females. Their average age was (44. 0 ± 1.2) (38 to 56) years old. Their injury was caused by a traffic accident (13 patients) and falling from a great height (2 patients). According to the Tile classification,there were 3 cases of type B1, 2 cases of type B2-2, 5 cases of type C1, 3 cases of type C2, and 2 cases of type C3. All the 15 patients were hemodynamically unstable and had a hypovolemic shock. Results There was a statistically significant increase in systolic blood pressure, hematocrit and hemoglobin measured immediately after EPP( P < 0. 05) . However, the units of RBC transfusion significantly decreased( P <0.05) and there were no significant difference in pulse rate( P > 0. 05) . The average operation time was (32.0 ± 2.6) min(30 to 40 min). Five patients died (72. 0 ± 4.2) h after operation ( 1 to 7 d). There was no significant difference between the survivors and the non-survivors in systolic blood pressure, pulse rate,hemoglobin, ISS score, hematocrit and blood transfusion( P > 0. 05). But there was significant difference in age and time between injury and EPP ( P < 0. 05). Conclusions In emergency treatment of patients with the hemodynamically unstable pelvic fracture, especially those combined with hypovolemic shock, EPP plus external fixation can effectively control the massive venous (presacral venous plexus) and bone hemorrhage.The blood pressure and pulse rate can be restored by fluid replacement and blood transfusion. Consequently,this method may significantly improve the hemodynamical parameters and decrease the mortality.

Objective To compare and analyze the biomechanical strengths of 4 different fixations for the posterolateral shearing tibial plateau fracture. Methods The posterolateral shearing fracture models were created in 40 synthetic tibia (SYNBONE, right) and randomly assigned into 4 groups. Group A was instrumented with 2 lag screws, group B with anteromedial LC-DCP, group C with lateral locking plate,and group D with posterolateral buttress plate. Vertical displacements of the posterolateral fragments and failure loads were measured under axial loads from 500 N to 1500 N. Results The vertical displacements in groups A, B, C and D were respectively(0. 459 ±0. 045) mm, (0. 365 ±0. 035) mm, (0. 264 ±0. 025)mm and (0. 128 ± 0. 018) mm under axial loads of 500 N, (1. 058 ± 0. 091 ) mm, (0. 882 ± 0. 053 ) mm,(0.551±0.053) mm and (0.440±0.068) mm under 1000 N, and (1.559±0.097) mm, (1.466±0. 079) mm, (1. 291 ±0. 077) mm and (0. 832 ±0. 130) mm under 1500 N. The differences between the 4 groups were statistically significant ( P ＜ 0. 05) . The loads to failure in groups A, B, C and D were respectively (1870 ± 156) N, (2520 ± 186) N, (2816 ± 190) N and (3465 ±210) N. The differences between the 4 groups were statistically significant ( P ＜ 0. 05 ). Conclusion The posterolateral buttress plate may be the biomechanically best fixation method for the posterolateral shearing tibial plateau fracture.

Objective The treatment of limb bone fracture in combination with main vessel injury wasanalyzed retrospectively.Methods Twenty ccsese were respectively conducted by interlocking intramedullary pin fixation,plate and screw fixation,plate and screw fixation orextemal brace fixation according to the injury conditions of the fractures.Then,vascahr injaries were flexibly dwelt with direct repair,end-to-end anastomosis or blood vessel grafting.Results The-limb save rate in this study is 70%.and the reaoons which csllsed the amputation included:Long ischemia time and serious tissue damage.Conclusion Fractures of the extremities with major vascular injuries should be diagnosed promptly to be conducted properly.In this way,better outcome would be obtained.

Animals ; Heme Oxygenase-1 ; biosynthesis ; Kidney ; drug effects ; metabolism ; Male ; Mercuric Chloride ; toxicity ; Oxidative Stress ; drug effects ; Random Allocation ; Rats ; Rats, Wistar

Adult ; Aluminum Silicates ; toxicity ; Humans ; Middle Aged ; Mining ; Occupational Exposure ; Prevalence ; Silicosis ; epidemiology

Objective T Iymphocytes were considered to be activated and involved in the ischemia-reperfusion injury during lung transplantation.Carbon dioxide pneumoperitoneum was shown to have inhibitory activity on the immune system.This study was designed to_investigate the effects of the effects of the therapeutic hypercapnia on the T Iymphocytes of rats in which ischemia-reperfusion injury occurred during lung transplantation.Melhods Sixteen Wismr rats weighed 300 to 400 g were randomized into control group(8 rats) or therapeutic group (8 ras)after transplantaion.Animals in both grotups were Oven inluded nitrogen(50%)and oxygen N2+(50%) at baseline. Animats in the control groap were given irked nitrogen (50%)and oxygen(50%)throughout the experiment ,and that in the thera-peutic group were given mixed gas which was composed of nitroged(40%),oxygen(60%)and carbon dioxide in appropriate concentra-tion to keep arterial partial pressure of carbon dioxide (PaCO2)at 80-100 mm Hg and FiO2 at 50%after reperfusion.All of the ani-mals were observed for 90 minutes after reperfusion.Mean arterial pressure(MAP) and arterila partial pressure of oxygen(PaO2) were recorded at baseline and every 15 minutes during the period of reperfusion.The expression of CD3,CD4 and CD28 in the peripheral blood was,examined,and the concentrations of Ifn-у,IL-2,IL-4 and IL-1O in the homogenate were measured after the experiment. Histological analysis of samples from transplanted lungs was performed.Resykts After reoerfysion,MAP and PaO2 in the therapeutic group were higher signitleantly than that in the group(P

Objective To investigate the effect of therapeutic hypercapnia on the inflammatory response in the rat lung transplantation. Methods Male pathogen free Wistar rats weighing 300-400 g were used in this study. The animals were anesthetized with 3% pentobarbital sodium 30 mg/kg, tracheostomized and mechanically ventilated (V_T 10 ml/kg, RR 50 bpm, FiO_2 50%). Carotid artery and femoral vein were cannulated for BP monitoring, blood sampling and fluid and drug administration. Left lung transplantation was performed using modified cuff technique. Forty-eight animals in which lung transplantation was successfully performed were randomized into 2 groups ( n = 24 each) : model group (M) and hypercapnia group (H) . In group H, PaCO_2 was maintained at 80-100 mm Hg by inhalation of CO_2.Arterial blood samples were obtained before lung transplantation (To , baseline) and at 1, 2, 4 h (T_(1-3)) of reperfusion for determination of blood TNF-α, IL-1 and IL-8 concentrations. The animals were then killed and the transplanted lungs were removed for microscopic examination and calculation of wet/dry lung weight ratio. Results The MAP and PaO_2 were significantly higher in group H than in group M. The blood IL-8 and TNF-α concentrations were significantly lower at T_(1-3) in group H than in group M, but there was no significant difference in blood IL-1 concentration between the 2 groups. The elastase content in the lung tissue was significantly lower at T_2 and T_3 in group H than in group M. Microscopic examination showed that the alveolar hemorrhage, the infiltration of the lung by macrophages and neutrophils and lung edema were significantly less in group H than in group M. Conclusion Therapeutic hypercapnia can obviously inhibit the inflammatory response in the rat lung transplantation.

Objective To construct the recombinant plasmid of fusion protein containing respiratory syncytial virus (RSV) cytotoxicity T lymphocyte (CTL) epitope M2:81-95 and heat shock protein (HSP) 70L1, and to investigate its immunogenicity after prokaryotic expression. Methods HSP70L1 gone was cloned from SMMC7721 cells. The M2:81-95 gene fragment was amplified by polymerase chain reaction (PCR) method. Plasmid pET-HSP70L1-M2 : 81-95 (pET-HSP70L1-M2) was constructed, identified and transferred into E. coli BL21 (DE3). Expression of HSP70L1-M2 : 81-95(HSP70L1-M2) was induced by isopropy-β-D-thiogalaetosidc (IPTG). The expressed protein was purified by affinity chromatography and renatured by gradient dialysis. The BALB/c mice were immunized with this fusion protein. IgG antibodies and the subtypes were detected by enzyme-linked immunosorbent assay (ELISA), and CTL responses were determined by methyl thiazolyl tetrazolium (MTT). Results The recombinant plasmid pET-HSP70L1-M2 was successfully constructed. The fusion protein HSP70L1-M2 was expressed in E. coll. The purified protein induced strong RSV-and CTL epitope-specific CTL responses and high titer of protein specific lgG antibody 4.87±0.35. The subtypes were IgG1 (5.53±0.28) and lgG2a (4.40±0.21) and IgG1/ IgG2a ratio was balanced. The titers of lgG, IgG1 and IgG2a in PBS control group were 0.33±0.17, 0.51±0.21 and 0, respectively, which werc significantly lower than those in immunized group (t = 3.512, 3.681, 5.856; P<0.05). Conclusions The recombinant plasmid pET-HSP70L1-M2 is successfully constructed and the fusion protein is expressed and purified. HSP70L1-M2 induced strong RSV-and CTL epitope-specific CTL responses and mixed T helper cell (Th)1/Th2 response in BALB/c mice.

AIM: To investigate the differential expression profile between nasopharyngeal carcinoma cell line CNE1 and its steady EBV-LMP1-transfected cell line CNE1-LMP1, and to explore the regulatory effect of LMP1 on oncomiRs expression in CNE1 cell line. METHODS: A microRNA array that targets 132 of the most well studied oncomiRs was used to detect the expression profile of CNE1 and CNE1-LMP1. qRT-PCR assay were used to verify the expression data detected by microarray. RESULTS: Among the restricted 132 miRNAs, 30 were detectable. Among which, 30 were expressed in CNE1-LMP1, 19 in CNE1 and 11 were specifically expressed in CNE1-LMP1. Among the 19 shared miRNAs, the expression level of 6 miRNAs (hsa-miR-19b, hsa-miR-17-3p, hsa-miR-22, hsa-miR-149, hsa-miR-150 and hsa-miR-188) elevated over two folds in CNE1-LMP1. No decrease in miRNA expression more than two folds was observed. qRT-PCR confirmed the expression difference of these six miRNAs (P<0.01). Among the 11 specifically expressed miRNAs in CNE1-LMP1, hsa-miR-122a showed the highest expression level surpassing the internal control sample. CONCLUSION: Our data suggest that LMP1 may play an important role in regulating the expression of miRNAs in tumor, which may be another important pathway employed by LMP1 in the development of nasopharyngeal carcinoma.