BACKGROUND: Osteoblasts have become a kind of important seed cells in bone tissue engineering. However, it is difficult to harvest osteoblasts, and the purity and calcification ability of osteoblasts isolated by different methods are inconsistent. OBJECTIVE: To compare the purity and calcification ability of osteoblasts induced from mouse bone marrow mesenchymal stem cells, MC3T3 cell lines, and cultured primarily from the neonatal mouse cranium. METHODS: Mouse bone marrow mesenchymal stem cells were isolated by differential adhesion method, and after passaing, passage 3 cells were cultured in osteogenic induction medium for 21 days. MC3T3 cell lines were cultured in osteogenic induction media 1 and 2 for 21 days. Osteoblasts were cultured primarily from neonatal mouse cranium by type Ⅰ coll agenase digestion method. Calcium nodules of osteoblasts obtained by three methods were observed by Alizarin red staining to detect osteogenic activity of cells. RESULTS AND CONCLUSION: (1) There were average 16.3 calcium nodules per low-power field after osteogenic induction of bone marrow mesenchymal stem cells. (2) There were sparsely distributed calcium nodules in MC3T3 cells after induction with osteogenic induction medium 1, accounting for 1.7 calcium nodules per low-power field, while there were dense calcium nodules in MC3T3 cells after induction with osteogenic induction medium 2, accounting for 44.6 calcium nodules per low-power field. There was a significant difference in the calcium nodule formation ability between the two groups (P < 0.01). (3) After primary culture, there was only 0.6 calcium nodule per low-power field. (4) Except for the insignificant difference between osteogenic induction medium 1 and primary culture groups, there were significant differences in pair-wise comparison of any other two groups. Except the insignificant difference between group I of MC3T3 inducing conditional media and primary culture osteoblasts, there were significant differences in the osteogenic ability between groups (P < 0.01). In conclusion, it is a better method to culture MC3T3 cells in osteogenic induction medium 2 containing dexamethasone, because many uncontrol able factors are involved in the isolation and culture of bone marrow mesenchymal stem cells.

Objective To evaluate the biological safety of 125I-filled carbon nanotubes covered with metallic esophageal stent with regard to the normal esophagus before clinical application.Methods 125I-filled carbon nanotubes covered with metallic esophageal stent was prepared.Eighteen of New Zealand rabbits were randomly divided into three groups with 6 rabbits in each group.Three groups of stents,non-radioactive,low radio-activity ( 3.7 - 5.6 MBq),and high activity ( 11.1 - 13.0 MBq ) were placed in the midpiece of esophagus of rabbits.Esophagus opacification and three-diamensions DSA were performed at 0.5 h,7,14 and 30 d after insertion of the stents,respectively.The rabbits were killed at 30d after insertion of the stents,and histologic examinations of the esophageal walls were performed.Results In non-radioactive and low activity groups,1 of 6 rabbits died of wound infection at 1 and 3 d after surgery due to pulmonary infection,respectively.All specimens were obtained from 16 rabbits.Microscopically,in all rabbits of low activity and high activity groups,there were membrana mucosa necrotic and swell and breakage of the muscle fiber in esophageal submucosa and muscularis,submucosal inflammation,which were more severe in high activity group.In low activity group,one esophagus ectal membrane was involved,however,esophageal perforation did not develop.In high activity group,3 of 6rabbits esophageal perforation had developed,in which one esophagus mediastinum fistula developed,without inflammation.In non-radioactive group,it was almost normal in mucosa layer,a small amount of inflammatory cells were found in submucosal layer,and part of muscle fibers was fractured and no pathological changes of necrosis was found.Conclusions Radioactive 125I carbon nanotubes covered metallic stent with low activity(3.7 -5.6 MBq) can be used as intraluminal palliative brachytherapy,which is safe and effective.

Objective To explore the incidence rate and the risk factors of deep vein thrombosis (DVT) in patients with traumatic fractures so as to provide references for prevention of DVT.Methods All of 534 Patients with fresh four extremities or pelvic fracture between January 2010 and December 2013 were involved in this study.The incidence of DVT under 5 risk factors including general state, injury type, fracture condition, operation and laboratory examination were analyzed.Each patient underwent three Doppler ultrasound exams in actions as the epidemiology diagnostic criterion for DVT.Results The total incidence rate of DVT in 534 patients was 11.99％.The univariate analysis showed that male patients with age≥60 years, BMI≥25 kg/m2, history of smoking, lack of exercises, history of diabetes, hypertension and coronary artery disease had higher incidence rate of DVT.In different injury types, the fall injury caused the highest incidence rate of DVT (45.71％).There were different DVT rates for different fracture sites, with the highest incidence rate of DVT for femur shaft fracture (20.69％).The incidence rate of DVT was 50.00％ for fractures of more than three parts, 15.29％ for fractures of two parts and only 3.98％ for sole part.The incidence rate of DVT for comminuted fractures was higher than others.The operation duration, massive transfusion during operation and general anesthesia were related with the increase of incidence of DVT.Positive ACA and enhancement of D-dimer, Fib and CRP were related with the increase of incidence of DVT.Conclusion The incidence of DVT in patients with traumatic fractures approaches a considerable level.It has relationships with age≥60 years, BMI≥25 kg/m2, history of smoking, fall injury, fracture of femoral shaft and hip, more than three parts of fractures, comminuted fractures, operation duration≥2 hours, largely blood transfused, massive transfusion during operation, general anesthesia, positive ACA, enhancement of D-dimer, Fib and CRP.The surgeons should recognize the importance to prevent DVT and PE in the traumatic patients.

Objective: To investigate the value of the trigeminocervical response (TCR) for revealing bulbar involvement in patients with spinal and bulbar muscular atrophy (SBMA). Methods: Thirty patients with SBMA and 30 healthy male controls were included in this study. In all of the normal controls, stimulation of the infraorbital nerve on one side produced bilateral short latency waves consisting of a positive/negative wave, p19/n31, the mean latency of which was measured. The mean square root of the ratio between the amplitude of p19/n31 and the mean rectifi ed surface electromyography (EMG) activity preceding the stimulus, the A value, was estimated. The parameters of the TCR were compared between the two groups. Results: Among the patients with SBMA, 21 (70.0%) had delayed latencies of p19/n31 (P < 0.01) and all (100%) had reduced A values (P < 0.01) relative to the normal controls. Conclusions: All parameters of the TCR were signifi cantly different between the patients with SBMA and the normal controls. T

Objective To evoke cerebral potentials by stimulating nociceptive fibers with contact heat evoked potentials stimulator (CHEPS)and estimate the nerve conduction velocities of peripheral nerve fibers mediating these responses.Methods Subjects were set in supine position.A heat-foil technology with a rapid rising speed at 70 ℃/s was used to elicit pain and contact heat evoked potentials(CHEP).Contact heat was delivered via one circular thermode (diameter 27 mm,area 573 mm~2).Thermal stimuli were sent at two intensity levels (49.5 ℃ and 54.5 ℃) to three body sites:thenar eminence,the dorsum of hand and proximal volar forarm.Contact heat evoked potentials were recorded from Cz and Pz.A systemic effect between stimulus intensities and pain rating were observed,the main components of this evoked potential were observed.Nerve conduction velocity was calculated from latency difference of CHEP and center to center distance of distal and proximal stimulus arrays.Results The pain intensity rating was 3.2?0.3 and 4.4?0.5 when thenar eminence was stimulated at the temperature of 49.5 ℃ and 54.5 ℃ respectively;the rating was 6.3?0.8 and 7.2?0.5 when the dorsum of hand and proximal volar forarm were stimulated at the temperature of 54.5 ℃ respectively.Three components,Cz/N550,Cz/P750 and Pz/P1000,were found in the evoked potentials.Nerve conduction velocities of the fibers were (12.9?7.5) and (1.7?0.4) m/s respectively,which were corresponding to those of A8 fiber and C fiber.Conclusions CHEPs can be elicited reliably and stably.Velocities of peripheral nerve fibers demonstrate that A8 fiber and C fiber mediate the response.

Objective To establish the method of contact heat evoked potential(CHEP)and to explore the value of this evoked potential in pain testing of patients with cerebral infarction.Methods A total of 100 healthy volunteers and 30 patients were examined.The healthy volunteers were divided into 3 groups according to the length of their arms:(Group A:56.0～65.0 cm ;Group B :65.5～74.0 cm ;Group C :74.5～83.0 cm).A recently de- veloped heat-foil technique with a rapid temperature rising rate at 70℃/s was used to elicit pain and contact heat e- voked potentials.Contact heat was delivered via one circular thermode(diameter 27 mm,area 573 mm~2)and set at two intensity levels(49.5℃and 54.5℃)to three body sites:the thenar eminence,the dorsum of hand and proximal volar forearm.The subjects were asked to rate the pain with numerical rating scale after each stimulus and CHEP was recorded from Cz and Pz.The association between stimulus intensities and pain rating was explored,the main compo- nents of the evuked potential were watched.CHEP,sensory conduction velocity(SCV)and somatosensory evoked potentials(SEP)were performed in patients with hemi-anesthesia caused by cerebral infarction.Results The pain intensity ratings were 3.2?0.3 and 4.4?0.5 at thenar eminence,5.0?0.7 and 6.3?0.8 at the dorsum of hand and 5.3?0.6 and 7.2?0.5 at the proximal volar forearm when the temperature of 49.5℃and 54.5℃was applied, respectively;Three components,Cz/N550,Cz/P750 and Pz/P1000,were identified in the evoked potentials.Cz/ N550 and Cz/P750 appeared when the dorsum of hand and proximal volar forearm were stimulated.In contrast,Pz/ P1000 could be identified when nociceptors of thenar eminence and proximal volar fbrearm were excited.In the pa- tients with cerebral infarction,CHEP disappeared or became abnormal on one side,while SCV and SEP were normal on that side.Conclusion It was suggested that CHEP could be elicited reliably in the controls.CHEP is helpful in the assessment of analgesia in patients with cerebral infarction.

Objective To observe the changes of plasma IL-8 levels and their association with the severity of and the passage of time after inhalation injury. Methods The plasma was separated from blood samples of 27 patients 12 h, 1, 2, 4 and7 d after inhalation injury. The IL-8 level in the plasma was determined by ELISA. Result IL-8 levels were significantly increased on day 1 and kept a high level for a period over 7 d. Conclusion Inhalation injury can induce the production of IL-8, and IL-8 level is positively correlated with the by severity of inhalation injury.

Objective To observe the changes of plasma IL-8 levels and their association with the severity of and the passage of time after inhalation injury. Methods The plasma was separated from blood samples of 27 patients 12 h, 1, 2, 4 and7 d after inhalation injury. The IL-8 level in the plasma was determined by ELISA. Result IL-8 levels were significantly increased on day 1 and kept a high level for a period over 7 d. Conclusion Inhalation injury can induce the production of IL-8, and IL-8 level is positively correlated with the by severity of inhalation injury.

Child ; Female ; Humans ; Lymphoma, Extranodal NK-T-Cell

Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0％,32.9％,39.0％ in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 ％,26.2 ％ ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.