Objective To investigate the expression of p16 gene in malignant mesothelioma,and its relationship with histological types,clinical stages and prognosis.Methods Immunohistochemical staining was used to detect the expression of p16 gene in the specimens of 80 cases of malignant mesothelioma.Results The positive expression of p16 gene was found in 35% malignant mesothelioma specimens,and negative in 65%(P

Objective To study the stability of anticoagulant peptide Hirulog-S and its lyophilized product, and to provide data on the storage conditions and clinical applications.Methods RP-HPLC was used to determine the content and the related substances of Hirulog-S and its lyophilized powder with influence factor test, accelerated test and long-term storage test.Results Light, temperature and humidity had no significant effect on the stability of Hirulog-S and its lyophilized powder in the influence factor test.The content and related substances of Hirulog-S and its lyophilized powder did not significantly change in the accelerated test ( 40℃, RH75%) and 24-month long-term storage test at room temperature and 4℃.Conclusion Hirulog-S and its lyophilized product are very stable, even after being stored at room temperature for two years.

A review of studies on the influence of impurities on protein crystallization is given.The possible sources of impurities and its effect on the protein crystallization are presented.The effects of impurities on protein crystallization,including nucleation,macroscopic morphologies,microscopic surface morphologies,growth rates,kinetics,quality,and repartitioning of impurities are reviewed.

Objective To compare the growth characteristics,proliferation and osteo/odontogenic differentiation capability of stem cells from human dental pulp (dental pulp stem cells,DPSCs) and apical papilla (stem cells from apical papilla,SCAP) in vitro.Methods Human dental pulp and apical papilla tissues were separated from impacted third molars of young healthy donors at the age of root development and digested by collagenase type Ⅰ and dispase type Ⅱ to derive primitive DPSCs and SCAP.Cells were then induced for osteo/odontongenic differentiation by medium containing β-glycerophosphate,dexamethasone and KH2PO4.Flow cytometry was utilized to test the expression of specific markers of stem cells,including CD24,CD34,CD45,CD90,CD105,CD146,STRO-1 and OCT-4.AR-S was used to display the mineralization structure and RT-PCR was applied to analyze the expression of bone sialoprotein (BSP),osteocalcin (OCN) and dentine sialophosphoprotein (DSPP).Results Both DPSCs and SCAP were positive for CD90,CD105,CD146,STRO-1 and OCT-4,in percentages varying according to cell type,without expression of CD34 or CD45.Only SCAP expressed CD24 positively.Both cells formed organized mineralization structure after 2 weeks of induction in time-dependent manner,with more mineralization by SCAP and expressed differentiation markers,including BSP,OCN and DSPP.Conclusion Human DPSCs and SCAP possess the characteristics of MSCs and could be differentiated into odontonblast-like cells in vitro.Both cells are approachable stem cell sources for dental tissue engineering.

Objective To comparatively study the characteristics of 3 kinds of culture substrates of human odontogenic induced pluripotent stem cells(iPSCs).Methods The human odontogenic iPSCs were cultured by 3 kinds of substrates:mouse embryonic fibroblasts(MEF),matrigel and recombinant human vitronectin(VTN-N).The iPSCs growth situation was compared among three groups.Results The preparation time of these 3 kinds of substrates was 14,3,1 hlespectively,and,the difference was statistically significant (P＜0.05).The iPSCs reprogramming time was (30± 1.6),(26 ± 2.1),(27 ± 1.4) d,lespectively,wht that in the MEF group significantly higer than in other two groups (P＜0.05).The reprogramming efficiencies were 0.3 ％ ± 0.03 ％,0.56 ％ ± 0.08 ％,0.7 ％ ± 0.02 ％ respectively (P＜ 0.05).Three kinds of substrate could better support iPSCs growth and make them to maintain un-differentiation status.Conclusion with no heterologous animal components,and the adrantaga of simple pleparation,oonfrollable standard and shorter gramming time is easy to prepare,the standard is controllable and the reprogramming time is shorter,which is an ideal substrate for supporting iPSCs growth.

BACKGROUNDp120 catenin (p120ctn) is an adheren junction protein that regulates barrier function, but its role has not been explored in alveolar edema induced by ventilation. We measured stretch-induced cell gap formation in MLE 12 cells due to the loss of p120. We hypothesized that alveolar permeability was increased by high lung inflation associated with alveolar epithelia cell tight junctions being destroyed, which resulted from the loss of p120.

METHODSCultured MLE12 cells were subjected to being stretched or un-stretched (control) and some cells were pretreated with pp2 (c-src inhibitor). After the end of stretching for 0, 1, 2, and 4 hours, the cells were lysed, and p120 expression and c-src activation was determined by Western blotting analysis. In vivo, SD rats were taken to different tidal volumes (Vt 7 ml/kg or 40 ml/kg, PEEP = 0, respiratory rate 30-40 betas/min) for 0, 1, 2, and 4 hour and some were pretreated with pp2, and alveolar edema was calculated.

RESULTSIt was found that p120 expression was reduced and c-src activation increased in a time-dependent and strain-dependent manner due to cyclic-stretch of the alveolar epithelial cells. These changes could be reversed by inhibition of c-src. We obtained similar changes in rats when they were subjected to large tidal volumes and the alveolar edema increased more than in rats in the low Vt group. Pretreated the rats with inhibition of c-src had less pulmonary edema induced by the high tidal volume ventilation.

CONCLUSIONSCyclic stretch MLE 12 cells induced the loss of p120 and may be the same reason by high tidal volume ventilation in rats can aggravate alveolar edema. Maintenance of p120 expression may be a novel therapeutic strategy for the prevention and treatment of ventilation induced lung injury (VILI).

Animals ; Blotting, Western ; Catenins ; physiology ; Cells, Cultured ; Mice ; Pulmonary Alveoli ; pathology ; Pulmonary Edema ; pathology ; Rats ; Rats, Sprague-Dawley ; Tidal Volume ; Ventilator-Induced Lung Injury ; pathology

ObjectiveTo investigate the relationship between the imaging manifestations and postoperative efficacy scores through measuring postoperative ankle mortise morphometry of Rüedi-Allg(o)wer Ⅲ pilon fracture.MethodsForty-seven cases of Rüedi-Allg(o)wer Ⅲ pilon fracture patients,including 20 males and 27 females with a mean of 43 years (range,21-65 years) were treated and followed up.Indexes including the width,height,depth,coronal angle and sagittal angle of ankle mortise were measured at the last follow-up.According to the Mazur score at the last follow-up,operated ankle functions were evaluated and divide into three groups (excellent group,good group and poor group.By comparing the five indexes of 47 cases and 3 groups between injury ankle and healthy ankle,we analyzed the correlation of the ankle mortise change and the ankle function.ResultsAll patients were followed up 18-24 months(mean,21 months).The results showed there were significant differences about all the measuring indexes except the height of ankle mortise between the injury ankle and healthy ankle of 47 follow-up cases.About the difference between both sides in 3 groups,the results showed there was a negative correlation between all indexes except the height of ankle mortise and the Mazur score.No correlation was found between the height of ankle mortise and the Mazur score.Recovering the ankle mortise's width,depth,coronal angle as well as sagittal anglein the surgery of severe pilon fractures has a significant effect on ankle function.ConclusionRecovery of the ankle mortise's width,depth,coronal angle as welld as sagittal angle during the surgery of severe pilon fractures has a significant effect on ankle function,so the anatomy of the ankle mortise should be recovered as much as possible.

BACKGROUND: Quantitative pharmaco-electroencephalography (QPEEG) can reflect cerebral cortical function, which can be certainly affected by general anesthetics. Anesthesia depth has good correlation with the anesthetic dosage, so if we can find out the areas of brain and band of QPEEG which is relative to the anesthetic dosage, the band may be taken as the index to reflect the depth of anesthesia. OBJECTIVE: To observe the effects of propofol on the alpha2-band (α2- band) of QPEEG in rabbits. DESIGN: A randomized control animal experiment. SETTING: Jiangsu Provincial Key Laboratory of Anesthesiology, Xuzhou Medical College. MATERIALS: The experiment was carried out in the animal laboratory of Xuzhou Medical College from October 2004 to August 2005. Thirtysix healthy adult rabbits were randomly divided into propofol 2.5, 5 and 10 mg/kg groups with 12 rabbits in each, including 6 were used to observe the change of percentage of each band power of QPEEG, and the other 6 were used to observe the latency and duration for the disappearance of righting reflex in the rabbits. METHODS: The experiment was performed between 14:00-17:00 every day. Rabbits in the three groups were treated with intravenous injection of propofol of 2.5, 5 and 10 mg/kg respectively within 30 seconds. ① The conscious rabbits were fixed onto the platform in a prone osition, and the QPEEG was recorded with the method of power spectrum analysis before administration and at 20, 30, 40, 50, 60, 70, 80, 90, 100 and 110 s and 2, 5, 10, 15, 20 and 30 minutes after administration respectively. The sampling time for each time point was 5 s. ② The latency and duration for the disappearance of righting reflex in the rabbits were recorded. RESULTS: ll the 36 rabbits were involved in the analysis of esults. ① After the intravenous injection of propofol, the righting reflexes all disappeared within 1 minute. The greater the dosage, the shorter the latency and the longer the duration r=0.79, P ＜ 0.01). ② Compared with before administration, propofol of 2.5 mg/kg had no obvious influence on the percentage of α2-band power (P ＞ 0.05); The percentages of α2-band power in the brain areas were increased after administration in the propofol 5 mg/kg group (P ＜ 0.05); Except that there were no significant differences in the left and right parietal regions between the propofol 10 mg/kg group and the propofol 5 mg/kg group, the percentages of α2-band power in the other brain areas in the propofol 10 mg/kg group were decreased as compared with those before administration and those in the other two groups (P ＜ 0.05), and the changes above were more obvious in the frontal and temporal regions.CONCLUSION: The influence of propofol on the percentage of α2-band power of QPEEG is biphasic, it is suggested that α2-band would be an index to reflect the anesthesia depth of propofol.

Objective Clutching internal fixtor(CIF)loose and the fixed part weakly heal up are often found in orthopedic clinic.In the present paper,biomechanics methods were used to try to explain and analyze these issues,providing a helpful suggestion for the application of CIF in clinic.Method Commerical finite element models(FEM)Program ANSYS was applied to set up the finite element models of orthopedic CIF and bone tissue to analyze and evaluate the biomechanical performances of the NiTi shape memory alloy CIF.Results There is an interaction force between embracing force of CIF and resistant force of bone tissue during the orthopedic clinical treatment.The embracing force along two semi-circular arms of CIF is increasing from the open position and reached the maximum value at the open symmetry position where the deformation of the bone occurred.Conclusion It is the key to choose the force loading position during the practical treatment,as the concentration force is the main force when there is an interactive force between the bone and the CIF.

AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein , and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell ( HLEC) membrane and the rabbit cornea.METHODS:The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR.The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identi-fied by Western blotting.PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC).The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested .RESULTS:The recombinant PTD-HSP27 plasmid was success-fully cloned and effectively expressed .The correctness of the recombinant protein PTD-HSP27 was demonstrated .Fluores-cence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs .Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor .CONCLUSION:The recombined gene PTD-HSPB1 was constructed by o-verlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatog-raphy column.Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cor-nea .