ABSTRACT
Objective: To explore the effect of natural hirudin on proliferation of human microvascular endothelial cells (HMVECs) and its preliminary mechanism of promoting angiogenesis. Methods: Three-dimensional culture models of HMVECs were established in vitro and observed by inverted phase contrast microscopy after 24 hours of culturing. Then, the three-dimensional culture models of HMVECs were treated with different concentrations (1, 4, and 7 ATU/mL) of the natural hirudin, respectively, and Dulbecco's modified Eagle's medium containing 10% fetal bovine serum as control. The cell proliferations of 4 groups were detected by cell counting kit 8 (CCK-8) method at 24, 48, and 72 hours; the angiogenesis of 4 groups were observed by tube formation assay at 24 hours; the expressions of vascular endothelial growth factor (VEGF) and Notch1 of HMVECs in 4 groups were observed by immunofluorescence staining at 24 hours. Results: The observation of cells in three-dimensional culture models showed that HMVECs attached to Matrigel well, and the cells formed tube structure completely after 24 hours. The results of CCK-8 test showed that the absorbance ( A) value of 1 and 4 ATU/mL groups were higher than that of control group at each time point ( P0.05). The results of immunofluorescence staining showed that compared with control group, the Notch1 expression was higher in 1 and 4 ATU/mL groups and lower in 7 ATU/mL group; and there was significant difference between 4 and 7 ATU/mL groups and control group ( P<0.05). The VEGF expression was higher in 1, 4, and 7 ATU/mL groups than in control group, in 4 ATU/mL group than in 1 and 7 ATU/mL groups, showing significant differences ( P<0.05). Conclusion: Natural hirudin can promote angiogenesis at low and medium concentrations, but suppress angiogenesis at high concentrations. Its mechanism may be related to the VEGF-Notch signal pathway.
ABSTRACT
BACKGROUND: Whether the differences exist between adipose-derived stem cells isolated from different parts of rats when cultured in vitro has been poorly understood. OBJECTIVE: To compare the growth characteristics and adipogenic ability of adipose-derived stem cells isolated from different parts of rats. METHODS: Freshly isolated adipose-derived stem cells were obtained from 5 mL inguinal groove and greater omentum adipose tissue of F344 rats using type Ⅰ col agenase digestion method. Then, adipose-derived stem cells were counted and cultured in vitro. Morphological and growth characteristics of adipose-derived stem cells derived from the two sites were observed. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was utilized to examine the doubling time of adipose-derived stem cells from different parts. The passage 2 adipose-derived stem cells were induced adipogenical y. Fourteen days after being induced, the differentiated cells were stained with oil red O and the positive cells were counted. The adipogenic differentiation ability of adipose-derived stem cells from the different parts was assessed. RESULTS AND CONCLUSION: (1) The number of adipose-derived stem cells from the greater omentum fat tissue in the same group was (281±10)×107/L, which was significantly higher than that from the inguinal groove fat tissue [(85±10)×107/L] (P < 0.01). Adipose-derived stem cells from the greater omentum and inguinal groove fat tissue achieved the exponential growth period on days 5 and 6, respectively, and achieved the platform period on days 9 and 10, respectively. The corresponding doubling time was 50 hours and 60 hours, respectively. After being passaged, adipose-derived stem cells grew in fibroblast-like shape actively. The adipogenic differentiation rate of adipose-derived stem cells from the greater omentum fat tissue was higher than that from the inguinal groove fat tissue [(38.90±2.86)% vs. (35.30±3.29)%, P < 0.01]. This shows that the number and the adipogenic differentiation ability of adipose-derived stem cells from different parts of the same F344 rat are different.
ABSTRACT
In order to observe the nutrition state in the severe multiple trauma patients undergoing adjuvant recombinant human growth hormone (rhGH) nutritional support therapy, 45 patients with severe multiple traumas (ISS＞25) were randomly divided into 3 groups. All the 3 groups had been supplied with nitrogen and caloricity according to the need of patients for 16 days. The rhGH therapy started 48 h after surgery and lasted for 14 days in two rhGH-treated groups in which rhGH was 0.2 and 0.4 U/(kg·d) respectively, and the resting group served as control one. The levels of nitrogen balance, prealbumin and safety variables (blood sugar, Na+, TT3 and TT4) were observed and compared among the three groups. The levels of nitrogen balance on the postoperative day (POD) 3 and 5 in the rhGH-treated groups were - 1.28±3.19, 5.45±2.00 and -0.18±2.55, 6.11±1.60, respectively,which were significantly higher than those in the control group (-5.17±1.68 and -1.08±3.31, P＜0.01). The values of prealbumin on the POD 3 and 5 in the rhGH-treated groups were 180.19±27.15, 194.44±50.82 and 194.94±29.65, 194.11±16.17, respectively, which were significantly higher than those in the control group (117.42±19.10 and 135.63±28.31, P＜0.01). There was no significant difference between the rhGH 0.2 U/(kg·d) group and rhGH 0.4 U/(kg·d) group in both of the levels of nitrogen balance and prealbumin. It is concluded that the nutritional support therapy with adjuvant rhGH which starts 48 h after surgery improves the nutrition state of the patients with severe multiple trauma. It is safe for severe multiple trauma patients who accept rhGH at the dose of 0.2 and 0.4 U/(kg·d).
ABSTRACT
In order to observe the nutrition state in the severe multiple trauma patients undergoing adjuvant recombinant human growth hormone (rhGH) nutritional support therapy, 45 patients with severe multiple traumas (ISS>25) were randomly divided into 3 groups. All the 3 groups had been supplied with nitrogen and caloricity according to the need of patients for 16 days. The rhGH therapy started 48 h after surgery and lasted for 14 days in two rhGH-treated groups in which rhGH was 0.2 and 0.4 U/(kg . d) respectively, and the resting group served as control one. The levels of nitrogen balance, prealbumin and safety variables (blood sugar, Na+, TT3 and TT4) were observed and compared among the three groups. The levels of nitrogen balance on the postoperative day (POD) 3 and 5 in the rhGH-treated groups were -1.28+/-3.19, 5.45+/-2.00 and -0.18+/-2.55, 6.11+/-1.60, respectively, which were significantly higher than those in the control group (-5.17+/-1.68 and -1.08+/-3.31, P<0.01). The values of prealbumin on the POD 3 and 5 in the rhGH-treated groups were 180.19+/-27.15, 194.44+/-50.82 and 194.94+/-29.65, 194.11+/-16.17, respectively, which were significantly higher than those in the control group (117.42+/-19.10 and 135.63+/-28.31, P<0.01). There was no significant difference between the rhGH 0.2 U/(kg . d) group and rhGH 0.4 U/(kg . d) group in both of the levels of nitrogen balance and prealbumin. It is concluded that the nutritional support therapy with adjuvant rhGH which starts 48 h after surgery improves the nutrition state of the patients with severe multiple trauma. It is safe for severe multiple trauma patients who accept rhGH at the dose of 0.2 and 0.4 U/(kg . d).