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 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 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 investigate the effect of methylprednisolone on the expression of glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) in Müller cells of rats’ retinae injured by laser. Methods Forty SD rats were randomly divided into two groups and inflicted with laser photocoagulation.The rats in treatment group were given methylprednisolone by intraperitoneal injection with a dose of 30 mg/kg for 3 days.At the 3rd,7th,14th,and 28th day after photocoagulation respectively, the eyes were enucleated,fixed and cut into sections.Immunohistochemical examination was used to detect the expression of PCNA and GFAP. Results After photocoagulation the Müller cells expressed PCNA both in the treatment and control group,and the expression of PCNA decreased sharply after 3 days. The expression of PCNA in treatment group was less than that in control group. After photocoagulation the Müller cells also expressed GFAP and the expression of GFAP lasted for at least 28 days ,and the expression of GFAP expression in the treatment group was less than that in the control group. Conclusion Methylprednisolone can reduce the expression of GFAP and PCNA in Müller cells of rats’ retinae injured by laser. (Chin J Ocul Fundus Dis, 2002, 18: 299-301)
Objective To investigate the effect of the injectable osteoinductive material with fibrin sealant(FS) as a carrier compounded with bone morphogenetic protein (BMP) on the proliferation and differentiation of marrow stromal cells (MSCs) towards osteoblasts and to provide the experimental foundation for the clinical application. Methods MSCs were extracted and cultured from bone marrow of the 3-day-old rabbit, and the third generation culturedMSCs were studied. The experiment included the experimental group(FS,including 1 μg/ml rhBMP-2), FS control group（FS）and blank control group (no material).The proliferation rate, the adhesive rate, the expression of the collagen Ⅰ and alkaline phosphatase, cell growth condition in the material and the ultrastructure of MSCs were investigated by electron microscopy, histochemistry and cell culture. Results The proliferation rate and the adhesive rate of MSCs in experimental group was significantly higher than those in blank control group ,but lower than those in FS control group （P＜0.05）. The expression level of thecollagen Ⅰ and alkaline phosphatase in the experimental group was significantlyhigher than those in all control groups（Marrow stromal cells Fibrin sealant Bone morphogenetic protein Cell culture Rabbits0.05）. Scanning electron microscope showed that the surface of material was rough and had many pores and that celland material mixed. Transmission electron microscope showed that MSCs of the experimental group were mostly of the phenotype of osteoblasts with relatively lowproliferation activity and high differentiation degree toward osteoblasts and with plenty of extracellular matrix and collagen fibers. MSCs of FS control group had low differentiation degree toward osteoblasts with few extracellular matrix and collagen fibers and high proliferation activity. MSCs of blank control group had low differentiation degree toward osteoblasts with few extracellularmatrix and collagen fibers, and low proliferation activity. Conclusion The injectable osteoinductive material with fibrin sealant as a carrier compounded with BMP could significantly accelerate the differentiation of MSCs towards osteoblasts. But it could not significantly accelerate the proliferation activity of MSCs.
ObjectiveTo systematically review efficacy application of fibrin glue (FG) after thyroidectomy.MethodsPubMed, EMbase, The Cochrane Library, ClinicalTrials.gov, CBM, CNKI, WanFang Data and VIP databases were searched to collect randomized controlled trials (RCTs) regarding the use of FG after thyroidectomy from inception to October 29th, 2019. Two reviewers independently screened literature, extracted data, and assessed the risk of bias of included studies. Then, meta-analysis was performed by using RevMan 5.3 software.ResultsA total of 15 RCTs involving 2 406 patients were included. The results of meta-analysis showed that compared with non-FG group, the use of FG could reduce postoperative drainage amount at the initial 24 hours (MD=?17.98, 95%CI ?28.35 to ?7.60, P=0.000 7), total amount of wound drainage (MD=?40.92, 95%CI ?46.25 to ?35.59, P<0.000 01), and postoperative discomfort (RR=0.48, 95%CI 0.35 to 0.66, P<0.000 01), as well as shorten drainage time (MD=?9.99, 95%CI ?15.74 to ?4.23, P=0.000 7) and stitches removal time (MD=?1.49, 95%CI ?2.1 to ?0.87, P<0.000 01). However, there was no statistically significant difference concerning postoperative short-term complications such as swelling (RR=0.78, 95%CI 0.48 to 1.28, P=0.32), recurrent laryngeal nerve injury (RR=0.83, 95%CI 0.21 to 3.29, P=0.79) and wound infection (RR=0.28, 95%CI 0.07 to 1.21, P=0.09) between two groups.ConclusionsThe current evidence shows that FG can reduce postoperative drainage amount and shorten postoperative recovery time in thyroidectomy. Due to the limited quality and quantity of included studies, more high quality studies are required to verify above conclusions.
In order to observe the effects of different facing directions of the germinal layer of periosteum on the cartilage regeneration, the human fibrin adhesive agent was used to adhere autogenous periosteum to repair the articular cartilage defect of rabbits. Twentyfour rabbits with 48 knee joints were divided randomly into two groups. A 0.6cm×1.2cm articular cartilage defect was created on the femoral trochlea until there was bleeding from the subchondral bone. A piece of periosteum, sized 0.75cm×1.5cm, was removed from the medial aspect of upper tibia. The periosteum was adhered to the defect by human fibrin adhesive agent. In Group 1 the germinal layer faced the subchondral bone and in Group 2 the germinal layer faced the joint cavity. The cartilage regeneration in both groups was observed by naked eyes and light microscope in 2nd and 6th weeks and by electron microscope after Safronin Ostained in 12th and 20th weeks. The results showed that before the 6th week, the cartilage regeneration was faster in Group 2 than that in Group 1. After that there was no significant difference in regeneration between the two groups. This suggested that the facing direction of the germinal layer was not a critical factor on cartilage regeneration. It was also found that the strength of the adhesive agent was not enough. The regenerated cartilage was proved to be hyaline cartilage.
【Abstract】ObjectiveTo investigate the changes and significances of the activity of blood coagulation and fibrinolysis in hepatectomy patients accompanied with chronic hepatic disease. MethodsThirtyfive patients who were accompanied with cirrhosis undertook surgery in the second affiliated hospital of Chongqing Medicall University from year 2003 to 2004 were divided into two groups: the first group of 18 cases received hepatectomy and the second group received nonhepatectomy surgical treatment. The (prothrombin time PT), (activated partial thromboplatin time APTT), (thrombin time TT), and the content of (Fibrinogen Fbg) and (Ddimer DD) in the blood drawn from peripheral veins were quantitatively measured by a fullyautomatic chromogenic and immunological assay machine (ACLFutura 9000,USA) at the phases of before operation, right after operation and 24hour after operation, respectively. ResultsAPTT in hepatectomy group increased significantly (P＜0.01) and were much higher than the nonhepatectomy group at corresponding phases (P＜0.01). PT in hepatectomy group increased even more significantly compared with that of preoperation and right after the operation (P＜0.01). The differences of TT at varying phases in hepatectomy group were of no significance (Pgt;0.05). There was also no significant difference of PT, APTT, and TT in nonhepatectomy group at varying phases. ConclusionThe function of blood coagulation is relatively poor and the secondary activity of fibrinolysis is overactivated in hepatectomy patients accompanied with chronic hepatic disease, which indicates a high risk of hemorrhage.
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.
The morbidity of coronary heart disease (CHD) is high, and the prognosis is unfavorable. Fibrinogen is both coagulation and inflammation factor, which has important influence on the occurrence and development of CHD. Previous studies reported that fibrinogen had relevance with traditional risk factors of CHD such as hypertension, diabetes and subclinical diseases such as left ventricular hypertrophy. The incidence of CHD increases with the fibrinogen level increasing. The fibrinogen level is higher in patients with CHD than that in healthy people. The coronary stenosis degree is heavier and the lesion is wider in patients with hyperfibrinogenemia. But the effects of fibrinogen on the secondary prevention of CHD is controversial. This paper summarized research progress based on the new understanding to fibrinogen on CHD recently.