Objective To compare the effects of heparin versus urokinase injection intrapleurally in the management of pleural thickening and adhesion due to tuberculous exudative pleurisy. Methods Sixty patients with tuberculous pleurisy were allocated into three groups randomly. Sodium heparin ( heparin group) , urokinase ( urokinase group) , and 0. 9% saline ( control group) were intrapleurally injected respectively. The concentrations of fibrinogen and D-dimer in pleural effusion were measured before and after the injection. The duration of absorption and the total drainage volume of pleural effusion were recorded. The pleural thickness and adhesion were observed two months after the injection. Results In 72 hours after the intrapleural injection, the concentration of fibrinogen( g/L) in the pleural effusion was significantly increased in the heparin group( 1. 13 ±0. 44 vs 0. 34 ±0. 19, P lt; 0. 001) , and significantly decreased in the urokinase group( 0. 25 ±0. 16 vs 0. 38 ±0. 15, P lt; 0. 05) when compared with baseline. Concentrations of D-dimer in the pleural effusions were significantly higher than those at baseline in both the heparin group and the urokinase group( 57. 0 ±17. 6 vs 40. 0 ±15. 4, P lt; 0. 05; 74. 5 ±16. 4 vs 43. 8 ±14. 9, P lt; 0. 001) . There were no significant differences in the absorption duration of pleural effusion among the three groups( P gt;0. 05) . The total drainage volume of pleural effusion was higher in the heparin group and the urokinase group compared to the control group( P lt;0. 01) . And the total volume of pleural effusion was significantly higher in the heparin group and the urokinase group than that in the control group( 2863 mL and 2465 mL vs 1828 mL,P lt;0. 01) . Two months after the intervention, the pleura were thinner[ ( 1. 37 ±0. 82) mm and ( 1. 33 ±0. 85) mmvs ( 3. 06 ±1. 20) mm, P lt; 0. 01] and the incidence of pleural adhesion was significantly lower[ 15% and 20% vs 50% , P lt; 0. 05] in the heparin and the urokinase groups than those in the control group.Conclusion Intrapleural heparin has similar effects with urokinase for prevention pleural thickness andadhesion in tuberculous pleurisy with good availability and safety.
Objective To investigate whether the sleep-induced hypoxemia ( SIH) at different time and different level have different effects on pulmonary emphysema and coagulation systemfunction in the rats with pulmonary emphysema. Methods Thirty Wistar rats were randomly divided into three groups( n = 10 in each group) . All rats were exposed to cigarette smoke twice a day ( 30 min each time) . From29th day on, the rats in Group A ( pulmonary emphysema with short SIH) were also exposed to mixed gas of 12. 5% oxygen for 1. 5 hours during sleeping time every day ( the expose time was divided into 4 periods, 22. 5 min each) . The rats in Group B ( pulmonary emphysema with mild SIH) were also exposed to mixed gas of 15% oxygen for three hours during sleeping time every day( the expose time was divided into 4 periods, 45 min each) . The rats in Group C( pulmonary emphysema with standard SIH) were also exposed to mixed gas of 12. 5% oxygen for three hours during sleeping time every day( the expose time was divided into 4 periods,45 min each) . After continuous exposure for 56 days, the rats were sacrificed. Semi-quantitative image analytic method was employed for histopathological analysis including pathological score of lungs, mean linear intercept ( MLI) and mean alveolus number( MAN) . ATⅢ, FIB, vWF, FⅧ were measured. Results All animals in three groups manifested the histopathological features of emphysema. Pathological scores of lungs and MLI of every group were significantly different from each other( F = 21. 907, F = 18. 415, all P lt; 0. 05) , Group A [ ( 61. 90 ±4. 25) % , ( 92. 45 ±1. 78) μm] and Group B[ ( 64. 60 ±3. 95) % , ( 92. 80 ±3. 65) μm] were significantly lower than Group C[ ( 73. 30 ±3. 86) % , ( 99. 32 ±2. 81) μm, q= 8. 96, q =6. 84, q = 12. 64, q =9. 65, all P lt; 0. 05] . Levels of FIB were significantly different among three groups ( F = 20. 592, P lt; 0. 05) while FIB in Group A[ ( 189. 98 ±5. 29) mg/ dL] and Group B[ ( 182. 70 ±2. 78) mg /dL] were significantly lower than that in Group C[ ( 198. 40 ±7. 37) mg/ dL, q = 4. 86, q= 9. 07, all P lt; 0. 05] , and FIB in Group A was significantly higher than that in Group B( q = 4. 20, P lt; 0. 05) . Levels of FⅧ were significantly different from each other( F = 33. 652, P lt;0. 05) while FⅧ in Group A[ ( 232. 26 ±4. 17) % ]and Group B[ ( 242. 53 ±14. 50) % ] were significantly lower than that in Group C[ ( 303. 25 ±32. 93) % ,q= 10. 73, q = 9. 18, all P lt; 0. 05] . Conclusions Pulmonary emphysema and hypercoagulable states increases with time and severity of SIH in rats with pulmonary emphysema. The elevated activity of blood coagulation factor may be a critical role in the hypercoagulable states.
Objective To determine if the levels of high-sensitivity C-reactive protein ( hs-CRP)and fibrinogen ( Fbg) can predict the risk of acute exacerbation of chronic obstructive pulmonary disease ( COPD) . Methods hs-CRP was measured by latex-enhanced immunoturbidimetric assay and Fbg was assessed by Von Clauss method. The number of exacerbations was recorded during a 6-month follow-up period. Results Fifty patients with stable COPD were enrolled in the study, of whom48 patients completed the trial and two patients dropped out. During the follow-up, 16 patients had once or more acute exacerbations while other 32 patients had no acute exacerbation. The patients were stratified into two groups ( A-exacerbation, B-no exacerbation) . At the baseline, the patients of the group A had lower FEV1 than thegroup B [ ( 1. 1 ±0. 4) L vs. ( 1. 4 ±0. 5) L, P lt;0. 05] . And the group A had higher hs-CRP and Fbg than the group B [ hs-CRP: ( 4. 6 ±3. 3) mg/L vs. 4. 3 mg/L( IQR 5. 5 mg/L) , P lt;0. 05] ; Fbg: ( 3. 8 ±0. 7) g/L vs. ( 3. 1 ±0. 5) g/L, P lt;0. 05] . Nine of 16 patients with a higher level of hs-CRP( hs-CRP gt;3 mg/L) had acute exacerbations. Seven of other 32 patients with normal hs-CRP level had acute exacerbations. The difference in the acute exacerbations rate between the two groups was significant ( 56. 25% vs. 21. 88% , P lt;0. 05) . All four patients with a higher level of Fbg( Fbg gt;4 g/L) had acute exacerbations. Twelve of 44 patients with normal Fbg level ( Fbg≤4 g/L) had acute exacerbations. The patients with Fbg more than 4 g/L had a higher rate of acute exacerbations( 100% vs. 27. 27%, P lt;0. 05) . After adjusting by age, bodymass index ( BMI) , FEV1 , tobacco consumption and other chronic diseases, the risk of acute exacerbation in individuals with baseline hs-CRP gt;3 mg/L was 9. 33 times higher than those with baseline hs-CRP≤3 mg/L ( 95% CI 1. 870-46. 573) . Conclusion Higher level of hs-CRP is associated with the high risk of exacerbation in patients with COPD.
Objective To investigate the division, prol iferation and differentiation abil ities of nestin+/GFAP+cell after spinal cord injury and to identify whether it has the characteristic of neural stem cells (NSCs). Methods Twelvemale SD rats, aged 8 weeks and weighing 200-250 g, were randomized into 2 groups (n=6 per group): model group inwhich the spinal cord injury model was establ ished by aneurysm cl ip compression method, and control group in which no processing was conducted. At 5 days after model ing, T8 spinal cord segment of rats in each group were obtained and the gray and the white substance of spinal cord outside the ependymal region around central tube were isolated to prepare single cellsuspension. Serum-free NSCs culture medium was adopted to culture and serum NSCs culture medium was appl ied to induce differentiation. Immunohistochemistry detection and flow cytometry were appl ied to observe and analyze the type of cells and their capabil ity of division, prol iferation and differentiation. Results At 3-7 days after injury, the model group witnessed a plenty of nestin+/GFAP+ cells in the single cell suspension, while the control group witnessed few. Cell count of the model and the control group was 5.15 ± 0.71 and 1.12 ± 0.38, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Concerning cell cycle, the proportion of S-phase cell and prol iferation index of the model group (15.49% ± 3.04%, 15.88% ± 2.56%) were obviously higher than those of the control group (5.84% ± 0.28%, 6.47% ± 0.61%), indicating there were significant differences between two groups (P lt; 0.01). In the model group, primary cells gradually formed threedimensional cell clone spheres, which were small in size, smooth in margin, protruding in center and positive for nestin immunofluorescence staining, and large amounts of cell clone spheres were harvested after multi ple passages. While in the control group, no obvious cell clone spheres was observed in the primary and passage culture of single cell suspension. At 5 days after induced differentiation of cloned spheres in the model group, immunofluorescence staining showed there were a number of galactocerebroside (GaLC) -nestin+ cells; at 5-7 days, there were abundance of β-tubul in III-nestin+ and GFAP-nestin+ cells; and at 5-14 days, GaLC+ ol igodendrocyte, β-tubul in II+ neuron and GalC+ cell body and protruding were observed. Conclusion Nestin+/GFAP+ cells obtained by isolating the gray and the white substance of spinal cord outside the ependymal region around central tube after compressive spinal cord injury in adult rat has the abil ity of self-renewal and the potential of multi-polarization and may be a renewable source of NSCs in the central nervous system.
Objective To compare the effect of two different methods of cell seeding on spatial distribution and gene expression of hBMSCs in biocoral scaffold in vitro cultures. Methods The composite of hBMSCs and biocoral scaffold was prepared by traditional seeding (group A) and fibrin glue seeding (group B). The seeding efficiency was measured after 30 minutes of incubation in group B and after 3 hours in group A. At 2, 7, 14 and 21 days after culture, the samples were harvestedand the serial longitudinal sections were cut for each embedded composite. The sections were stained with DAPI and were measured using fluorescence microscope with apotome under serial optical sections. The cell number in every 10 × objective field was automatically measured by AxioVision image analysis software and levels (from seeding surface to bottom L1-L5) or columns (from centre to margin) for comparing cell distribution were set up. The specific osteogenic genes [osteonectin (ON), core binding factor α1 (Cbfα1), osteocalcin (OC)] expression was measured by RT-PCR. Results The seeding efficiency was significantly higher in group B (88.32% ± 4.2%) than in group A (66.51% ± 12.33%, P lt; 0.01). At 2 days after culture, the cell number from L1 to L4 decreased gradully in two groups (P lt; 0.05); in the cell number of different columns, there was no significant difference in group A (Pgt; 0.05) whereas significant difference in group B (P lt; 0.05); there was no significant difference in gene expression between two groups (P gt; 0.05). At 7 days after culture, the cell number was less than that at 2 days in group A and there was significant difference among levels (P lt; 0.05). The cell number and osteogenic gene expression increased sharply and there appeared uniform cell distribution in group B (P gt; 0.05). The gene expression of ON and Cbfα1 in group B was higher than that in group A (Plt; 0.05). At 14 days after culture, the cell number in levels or columns in group A decreased sharply and was less than that at 7 days (P lt; 0.05); whereas the cell number was similar to that at 7 days in group B (P gt; 0.05). The OC gene expression reached the highest level in group B at 14 days. The gene expression was higher in group B than in group A (P lt; 0.05). At 21 days after culture, there was significant difference in the cell number among levels and in the gene expression between group A and group B (P lt; 0.05); there was no significant difference in the cell number among columns in two groups (Pgt; 0.05). In addition, the cell number of most levels and columns in group B was more than that in group A at 7, 14 and 21 days after culture (P lt; 0.05). Conclusion More uniform cell distribution with rapid prol iferation and osteogenic differentiation is available in different levels or columns of scaffold by fibrin glue seeding than by traditional seeding.
Objective To analyze MC3T3E1 cell morphology, prol iferation, and osteogenic differentiation in fibrin gel (FG) so as to lay a fundament for use of FG in tissue engneering. Methods MC3T3E1 cells were incubated in three concentrations (20, 10 and 5 mg/mL)of FG as the experimental groups (groups A, B and C) and in the common medium culture as the control group (group D). The cell morphology and distribution in FG were observed by inverted phase contrast microscope and confocal laser scanning microscope at different time. The cell prol iferation was assessed by fluorospectrophotometer. The alkal ine phosphatase (ALP) activity was detected by automatic biochemistry analyses and von Kossa staining was used to analyze calcium salts mineralization. RT-PCR was used to analyze the ALP and bone sialoprotein (BSP)mRNA expression at 14 and 21 days. Results In groups A, B and C, the MC3T3E1 cells had long processes which connected each other and formed network; but fusiform or cube cells were observed in group D at 21 days. The fluorescence intensity was increased gradually with time, was the highest at 14 days and the lowest at 28 days in group D; it was highest in groups A, B and C at 28 days, there were statistically significant differences when compared with group D (P lt; 0.05). The ALP activity was increased gradually with time, and it was the highest at 28 days in group D and at 21 days in groups A and B, there were significant differences (P lt; 0.05), no statistically significant differences compared with group D at other time points (P gt; 0.05). The mineral ization nodus were seen at 21 and 28 days in group A, but no mineral ization nodus was seen in group D at 28 days. The RT-PCR results showed the mRNA expressions of ALP and BSP were enhanced in group A when compared with group D (P lt; 0.05). Conclusion The osteogenic differentiation was most obvious and cell prol iferation was most active after 21 days of incubation in FG.
【Abstract】 Objective To broaden the cl inical uses of fibrin-based biomaterials and to develop further study incell biology and to comprehensively understand and master related knowledge with regard to the present development status of fibrin. Methods Many relevant domestic and international papers were reviewed to make a summary. Results Recognization was obtained from four aspects, which were structure and function of fibrinogen, cl inical use of fibrin, fibrin scaffold for tissue engineering, and compounding biomaterials of fibrin. It showed that every aspect had great research extension and practical appl ication. Conclusion Besides a surgical hemostat and sealant, fibrin has great potentials in playing roles of tissue engineering scaffold, drug del ivery vehicle, and compounding material.