OBJECTIVETo establish a convenient and accurate method of DNA molecular marker for the identification of corium stomachium galli.

METHODCytb mtDNA sequences of Gallus gallus domestica and three other species of poultry were downloaded from Genbank. Species-specific PCR primers were designed according to the differential DNA fragments of the cytb genes. PCR tests were performed with the DNAs extracted from G. gallus domestica, three other poultry species and domestic mammal animals.

RESULTThe specific primers of G. gallus domestica could only amplify the cytb mtDNA of G. gallus domestica.

CONCLUSIONThe primers are specific to G. gallus domestica mtDNA and can used to discern Corium stomachium from the false medicine.

Animals ; Chickens ; classification ; genetics ; DNA, Mitochondrial ; genetics ; Liver ; metabolism ; Polymerase Chain Reaction

Objective:To explore the effect of distal veins on the survival of a dorsal four-territory perforator flap in rats.Methods:A total of 32 SD rats were randomly divided into the control group and the experimental group, with 16 rats in each group. The multi-territory perforator flap including the bilateral iliolumbar and bilateral posterior intercostal angiosomes was cut from the back of each rat, with the size of 6 cm×7 cm. The right iliolumbar artery and vein were preserved in the control group, while the right iliolumbar artery and the right posterior intercostal vein were preserved in the experimental group. In both groups, incisions were made between the right iliolumbar angiosome and the right posterior intercostal angiosome. Finally, the flap was sutured back to their orthotopic site. At 6 hours and 1, 3, 5, 7 days after surgery, the blood perfusion at the bilateral iliolumbar and the left posterior intercostal vascular territories were measured. On the seventh day after surgery, the percentage of the survived area of the flaps were evaluated, arteriography was performed to observe the dilation of arteries within the flap, the intraluminal diameter of the choke artery in the choke 2 area was measured using hematoxylin and eosin staining, and the relative expression of endothelial nitric oxide synthase (eNOS) was detected by Western blotting. SPSS 28.0 was used for statistical analysis, and measurement data were presented as Mean±SD. Independent sample t-test was used to compare data across two groups. P<0.05 was considered statistically significant. Results:(1) At 6 hours and 1, 3, 5, 7 days after surgery, the experimental group displayed higher blood perfusion than the control group at the bilateral iliolumbar and the left posterior intercostal vascular territories (all P<0.01). (2) On the 7th day after surgery, the artery dilation of the experimental group was more obvious than that of the control group; the percentage of the survived flap area in the experimental group was higher than that in the control group [(87.6±3.2)% vs. (65.3±3.0)%, P<0.01]; the intraluminal diameter of the choke artery was greater in the experimental group than that in the control group[(49.3±3.1) μm vs. (35.1±2.3) μm, P<0.01]; and the relative expression of eNOS in the experimental group was higher than that in the control group (0.87±0.07 vs. 0.50±0.05, P<0.01). Conclusion:The distal vein (right posterior intercostal vein) of dorsal four-territory perforator flap of SD rats directly guided the pedicle artery blood supply to promote the expression of eNOS, dilated the arteries in each zone of the flap, increased the blood supply to the distal artery of the flap, and ultimately enhanced the flap survival area.

Objective:To compare the effects of leukocyte-poor platelet-rich plasma (Lp-PRP) and leukocyte-rich platelet-rich plasma (Lr-PRP) on dorsal wound healing in rats.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into Lp-PRP group, Lr-PRP group and control group, each containing twelve rats. Venous blood was drawn and the Lp-PRP and Lr-PRP were prepared separately using a centrifugal method. Circular full-thickness skin defect wounds (15 mm in diameter) were created on the backs of the rats in the three groups. The wounds were then treated with 100 μl Lp-PRP, Lr-PRP and saline, respectively. At 7 and 14 days post-operation, the wounds were photographed, and Image J software was used to calculate the wound area rate (postoperative wound area/wound area at modeling time × 100%). At 14 days post-operation, the total neo-epithelium length and collagen deposition rate of the wounds were evaluated using HE and Masson staining, respectively. At 7 days post-operation, the relative expression of vascular endothelial growth factor (VEGF) in the wounds was detected by Western blotting, and the number of CD31 positive microvessels in the wounds was examined by immunohistochemistry. Statistical analysis was performed using SPSS 28.0. One-way analysis of variance (ANOVA) was used to compare the three groups, and Tukey’s test was used for pairwise comparisons. A significance level of P<0.05 was considered statistically significant. Results:Blood analysis revealed that the platelet concentrations in the prepared Lp-PRP and Lr-PRP were 4.1 times and 4.5 times that of whole blood, respectively ( P<0.01), with no significant difference between the two PRPs ( P>0.05). The leukocyte concentration in Lp-PRP was undetectable, while in Lr-PRP, it was 3.5 times that of whole blood ( P<0.01). The wound area rate at 7 and 14 days post-operation, the total neo-epithelium length and collagen deposition rate at 14 days post-operation, as well as the relative expression of VEGF and the number of CD31-positive microvessels at 7 days post-operation in the Lp-PRP and Lr-PRP groups were superior to those in the control group (all P<0.01). However, there was no significant difference between the two PRP groups (all P>0.05). Conclusion:Both Lp-PRP and Lr-PRP promote dorsal wound healing in rats by enhancing re-epithelialization, collagen deposition, and angiogenesis. The impacts of Lp-PRP and Lr-PRP on promoting wound healing are comparable and not influenced by the presence of leukocytes in PRPs.

Objective:To explore the effect of distal veins on the survival of a dorsal four-territory perforator flap in rats.Methods:A total of 32 SD rats were randomly divided into the control group and the experimental group, with 16 rats in each group. The multi-territory perforator flap including the bilateral iliolumbar and bilateral posterior intercostal angiosomes was cut from the back of each rat, with the size of 6 cm×7 cm. The right iliolumbar artery and vein were preserved in the control group, while the right iliolumbar artery and the right posterior intercostal vein were preserved in the experimental group. In both groups, incisions were made between the right iliolumbar angiosome and the right posterior intercostal angiosome. Finally, the flap was sutured back to their orthotopic site. At 6 hours and 1, 3, 5, 7 days after surgery, the blood perfusion at the bilateral iliolumbar and the left posterior intercostal vascular territories were measured. On the seventh day after surgery, the percentage of the survived area of the flaps were evaluated, arteriography was performed to observe the dilation of arteries within the flap, the intraluminal diameter of the choke artery in the choke 2 area was measured using hematoxylin and eosin staining, and the relative expression of endothelial nitric oxide synthase (eNOS) was detected by Western blotting. SPSS 28.0 was used for statistical analysis, and measurement data were presented as Mean±SD. Independent sample t-test was used to compare data across two groups. P<0.05 was considered statistically significant. Results:(1) At 6 hours and 1, 3, 5, 7 days after surgery, the experimental group displayed higher blood perfusion than the control group at the bilateral iliolumbar and the left posterior intercostal vascular territories (all P<0.01). (2) On the 7th day after surgery, the artery dilation of the experimental group was more obvious than that of the control group; the percentage of the survived flap area in the experimental group was higher than that in the control group [(87.6±3.2)% vs. (65.3±3.0)%, P<0.01]; the intraluminal diameter of the choke artery was greater in the experimental group than that in the control group[(49.3±3.1) μm vs. (35.1±2.3) μm, P<0.01]; and the relative expression of eNOS in the experimental group was higher than that in the control group (0.87±0.07 vs. 0.50±0.05, P<0.01). Conclusion:The distal vein (right posterior intercostal vein) of dorsal four-territory perforator flap of SD rats directly guided the pedicle artery blood supply to promote the expression of eNOS, dilated the arteries in each zone of the flap, increased the blood supply to the distal artery of the flap, and ultimately enhanced the flap survival area.

Objective:To compare the effects of leukocyte-poor platelet-rich plasma (Lp-PRP) and leukocyte-rich platelet-rich plasma (Lr-PRP) on dorsal wound healing in rats.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into Lp-PRP group, Lr-PRP group and control group, each containing twelve rats. Venous blood was drawn and the Lp-PRP and Lr-PRP were prepared separately using a centrifugal method. Circular full-thickness skin defect wounds (15 mm in diameter) were created on the backs of the rats in the three groups. The wounds were then treated with 100 μl Lp-PRP, Lr-PRP and saline, respectively. At 7 and 14 days post-operation, the wounds were photographed, and Image J software was used to calculate the wound area rate (postoperative wound area/wound area at modeling time × 100%). At 14 days post-operation, the total neo-epithelium length and collagen deposition rate of the wounds were evaluated using HE and Masson staining, respectively. At 7 days post-operation, the relative expression of vascular endothelial growth factor (VEGF) in the wounds was detected by Western blotting, and the number of CD31 positive microvessels in the wounds was examined by immunohistochemistry. Statistical analysis was performed using SPSS 28.0. One-way analysis of variance (ANOVA) was used to compare the three groups, and Tukey’s test was used for pairwise comparisons. A significance level of P<0.05 was considered statistically significant. Results:Blood analysis revealed that the platelet concentrations in the prepared Lp-PRP and Lr-PRP were 4.1 times and 4.5 times that of whole blood, respectively ( P<0.01), with no significant difference between the two PRPs ( P>0.05). The leukocyte concentration in Lp-PRP was undetectable, while in Lr-PRP, it was 3.5 times that of whole blood ( P<0.01). The wound area rate at 7 and 14 days post-operation, the total neo-epithelium length and collagen deposition rate at 14 days post-operation, as well as the relative expression of VEGF and the number of CD31-positive microvessels at 7 days post-operation in the Lp-PRP and Lr-PRP groups were superior to those in the control group (all P<0.01). However, there was no significant difference between the two PRP groups (all P>0.05). Conclusion:Both Lp-PRP and Lr-PRP promote dorsal wound healing in rats by enhancing re-epithelialization, collagen deposition, and angiogenesis. The impacts of Lp-PRP and Lr-PRP on promoting wound healing are comparable and not influenced by the presence of leukocytes in PRPs.