Purpose:To determine the efficacy of a low-molecular-weight heparin (fraxipparine) administered in the infusion fluid to prevent postoperative fibrin formation in a rabbit lensectomy and vitrectomy model.
Methods:Fourteen adult pigmented rabbits were randomly as signed into two groups.Standard fragmatome lensectomies and core vitrectomies were performed prospectively in a masked fashion on control eyes with balanced salt infusion and on experimental eyes treated with fraxiparine/ml 6U in the infusate.Intraoperative bleeding was graded in a masked fashion by the surgeon. The amounts of firbrin and hemorrhage were graded in a masked fashion on postoperative days 1 through 7.
Results:The mean grde of fibrin in the eyes treated with fraxiparine was lower than that in the control eyes (Plt;0.01) on postoperative days 1 through 3 respectively.Also,the average days to clear the fibrin in the eyes treated with fraxiparine was shorter than that in the control eyes (P＝0.001).No statistically significant differences in the degree of intraoperative or postoperative hemorrhage were noted between the two groups.
Conclusion:Low-molecular weight heparin(fraxiparine) is an effetive inhibit or of postoperative fibrin formation in a rabbit model and is not associated with is not associated with an increased risk of intraoperative or postopertive bleeding at the tested dose.
(Chin J Ocul Fundus Dis,1998,14:35-37)
Objective To investigate the effect of local delayed releasing vascular endothelial growth factor (VEGF) on accelerating healing of intestinal anastomotic stoma. Methods An intra-abdominal infection modal of rabbit was established by artificial appendix perforation, and excision and anastomosis of terminal ileum were subsequently performed after 12 h. The animals were divided into four groups (32 for each group) with different reagents on anastomotic surface: control group, fibrin glue group (FG group), VEGF group, and VEGF＋FG group. The incidence of stomal leak, anastomosic bursting pressure, hydroxyproline content, and expression of VEGF in cured stoma tissue were measured respectively on day 3, 5, 7 and 14 after operation. Results The total incidence rate of leakage was lower in FG group and VEGF group than that in control group, but there was no statistical significance （Pgt;0.05）. The incidence rate was significantly lower in FG＋VEGF group than that in control group （Plt;0.05）. On day 14 postoperatively, the bursting pressure of anastomotic stoma, hydroxyproline content, and positive cell expression rate of VEGF protein (except VEGF group) were significantly increased in FG＋VEGF group than those in other three groups （Plt;0.05, Plt;0.01）. Conclusion Local delayed release of VEGF by fibrin glue can improve the healing of intestinal anastomotic stoma and reduce the incidence of stomal leak.
Objective To study the relationship between the levels of plasma fibrinogen, serum bilirubin, uric acid and mild stenosis of coronary artery. Methods Patients with suspected myocardial ischemia who underwent coronary angiography in our hospital were divided into the coronary artery mild stenosis group and the normal control group according to the result of coronary arteriongraphy between April 2007 to May 2009. Logistic regression was used to identify the risk factor of mild stenosis of coronary artery. Results Two-hundred and seventy-nine patients involving 191 patients with mild stenosis of coronary artery and 88 patients with normal coronary artery were included. The factors of gender, age, history of hypertension or diabetes mellitus, smoking history, systolic blood pressure, levels of plasma fibrinogen and serum creatinin were significantly different between the two groups. Multivariate logistic regression models found that the factors of age (OR=1.084, 95%CI 1.040 to 1.129, Plt;0.001), hypertension (OR=3.025, 95%CI 1.462 to 6.261, P=0.003), diabetes mellitus (OR=2.519, 95%CI 1.066 to 5.951, P=0.035), smoking history (OR=5.412, 95%CI 2.186 to 13.401, Plt;0.001), plasma fibrinogen (OR=1.748, 95%CI 1.059 to 2.885, P=0.029), serum bilirubin (OR=0.599, 95%CI 0.418 to 0.858, P=0.005), and high-density lipoprotein (HDL) cholesterol (OR=0.219, 95%CI 0.049 to 0.985, P=0.048) were independently associated with mild stenosis of coronary artery. By contrast, the level of serum uric acid was not associated with mild stenosis of coronary artery. Conclusion Except for traditional risk factors, levels of fibrinogen and bilirubin are independent risk factors of mild stenosis of coronary artery.
The fundus lesions caused by high myopia (HM) often lead to irreversible visual impairment or even blindness. However, the pathogenesis of HM and its fundus lesions is still unclear, the intraocular fluid detection technology of micro samples has brought new prospects for the early diagnosis, monitoring and intervention of the fundus lesions. The molecules associated with HM are various and functionally diverse, intermolecular interactions are staggered and the specific mechanism is complex. With the development of intraocular fluid detection technology, while gradually revealing the role of each molecule in the pathogenesis of HM, it is expected to successfully assist clinical work in the future, providing outpost markers for the progress of myopia and targets for early intervention, or providing a new therapy choice for HM fundus lesions at the molecular level targeting pathogenesis, which is expected to provide more accurate and effective treatment for HM patients in the future.
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 determine the influence and significance of combinative assessment of 64 multi-slice spiral computer tomography (MSCT) with serum amyloid A protein (SAA) or fibrinogen (FIB) on the selection of operative procedures of rectal cancer under the multidisciplinary team. Methods Prospectively enrolled 240 patients diagnosed definitely as rectal cancer at West China Hospital of Sichuan University from February to June 2009 were randomly assigned into two groups. In one group named MSCT+SAA group, both MSCT and SAA combinative assessment were made for the preoperative evaluation. In another group named MSCT+FIB group, both MSCT and FIB combinative assessment were made for preoperative evaluation. Furthermore, the preoperative staging and predicted operation procedures were compared with postoperative pathologic staging and practical operation procedures, respectively, and the relationship between the choice of operation procedures and clinicopathologic factors was analyzed. Results According to the criteria, 234 patients were actually included into MSCT+SAA group (n=118) and MSCT+FIB group (n=116). The baseline characteristics of two groups were statistically similar (Pgt;0.05). For MSCT+SAA group, the accuracies of preoperative staging T, N, M and TNM were 72.9%, 83.1%, 100% and 80.1%, respectively. For MSCT+FIB group, the accuracies of preoperative staging T, N, M and TNM were 68.1%, 75.0%, 100% and 74.1%, respectively, and there was not a statistically significant difference (Pgt;0.05). There was also not a statistically significant difference of the accuracy of prediction to operative procedures in two groups (99.6% vs. 96.6%, Pgt;0.05). The preoperative T staging (P＜0.001), N staging (P＜0.001), TNM staging (P＜0.001), serum level of SAA (P＜0.001), serum level of FIB (Plt;0.001) and distance of tumor to the dentate line (P＜0.05) were associated to the operative procedures. Conclusions Combinative assessment of MSCT and FIB could improve the accuracy of preoperative staging and operative procedures prediction, however, it may be not superior to MSCT plus SAA.
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.
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.
ObjectiveTo explore the function of intercellular adhesion A (icaA), fibrinogen binding protein (fbe), and accumulation-associated protein (aap) genes in formation of Staphylococcus epidermidis-Candida albicans mixed species biofilms.
MethodsThe experiment was divided into 3 groups:single culture of Staphylococcus epidermidis ATCC35984 (S. epidermidis group) or Candida albicans ATCC10231 (C. albicans group), and co-culture of two strains (mixed group) to build in vitro biofilm model. Biofilm mass was detected by crystal violet semi-quantitative adherence assay at 2, 4, 6, 8, 12, 24, 48, and 72 hours after incubation. XTT assay was performed to determine the growth kinetics in the same time. Scanning electron microscopy (SEM) was used to observe the ultrastructure of the biofilms after 24 and 72 hours of incubation. The expressions of icaA, fbe, and aap genes were analyzed by real-time fluorescent quantitative PCR.
ResultsCrystal violet semi-quantitative adherence assay showed that the biofilms thickened at 12 hours in the S. epidermidis and mixed groups; after co-cultured for 72 hours the thickness of biofilm in mixed group was more than that in the S. epidermidis group, and there was significant difference between 2 groups at the other time (P<0.05) except at 72 hours (P>0.05). In C. albicans group, the biofilm started to grow at 12 hours of cultivation, but the thickness of the biofilm was significantly lower than that in the mixed group in all the time points (P<0.05). XTT assay showed that the overall growth speed in the mixed group was greater than that in the C. albicans group, and it was greater than that in the S. epidermidis group at 48 hours; there was no significant difference in the growth speed between the mixed groups and the S. epidermidis group in the other time points (P>0.05) except at 12 hours (P<0.05). The absorbance (A) value in the mixed group was lower than that in the S. epidermidis group at 2 and 4 hours, but no significant difference was shown (P>0.05); the A value of mixed group was significantly higher than that of the C. albicans group after 6 hours (P<0.05). SEM observation showed that mature biofilms with complex structure formed in all groups. The real-time fluorescent quantitative PCR showed the expressions of fbe, icaA, and aap genes in mixed group increased 1.93, 1.52, and 1.46 times respectively at 72 hours compared with the S. epidermidis group (P<0.05).
ConclusionMixed species biofilms have more complex structure and are thicker than single species biofilms of Staphylococcus epidermidis or Candida albicans, which is related to increased expressions of the icaA, fbe, and aap genes of Staphylococcus epidermidis.
Objective To establish the optimal morphological criteria combined with fibrinogen level for evaluation of lymph node metastasis in colorectal cancer. Methods A consecutive series of 690 patients who underwent curative surgery for colorectal cancer, were examined by abdominopelvic enhanced multi-slice spiral computed tomography (MSCT) scan. If regional lymph nodes appeared, the maximal long-axis diameter (MLAD), maximal short-axis diameter (MSAD), and axial ratio (MSAD/MLAD) were recorded. At each lymph node size cut-off value, the following were calculated: accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Moreover, preoperative plasma level of fibrinogen was retrospectively examined to identify metastatic or inflammatory lymph node combined with MSCT image. Both modalities, MSCT plus fibrinogen and MSCT alone, were compared based on the pathologic findings. Results The study population consisted of 100 patients with regional lymph nodes show. No significant difference was found between metastatic and inflammatory lymph nodes in imaging characteristics (Pgt;0.05). The best cut-off value of MSAD was 6 mm for lymph node metastasis with the sensitivity of 46.8%, specificity of 68.4%, accuracy of 55.0%, PPV of 70.7% and NPV of 44.1%. The best cut-off value of MLAD was 8 mm with the sensitivity of 43.5%, specificity of 63.2%, accuracy of 51.0%, PPV of 65.9% and NPV of 40.7%. Using hyperfibrinogenemia (FIB ≥3.5 g/L) to identify small metastatic lymph node, of which MSAD lt;6 mm or MLAD lt;8 mm, showed statistical diagnostic value (Kappa=0.256, P=0.047). Compared with MSAD (6 mm) alone, MSAD (6 mm) combined with hyperfibrinogenemia had a higher sensitivity (79.0% vs. 46.8%, Plt;0.001), but a similar accuracy (66.0% vs. 55.0%, Pgt;0.05) and a lower specificity (44.7% vs. 68.4%, P=0.037). MLAD (8 mm) combined with hyperfibrinogenemia led to a greater diagnostic value in sensitivity (80.6% vs. 43.5%, Plt;0.001) and accuracy (66.0% vs. 51.0%, P=0.031) than MLAD (8 mm) alone, with a no-significantly decreasing specificity (42.1% vs. 63.2%, Pgt;0.05). Conclusions This present study recommend MSAD ≥6 mm or MLAD ≥8 mm as the optimal criteria for preoperative N staging in colorectal cancer. Moreover, the sensitivity and even accuracy could be improved by combining hyperfibrinogenemia for lymph node metastasis identification.
