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Normal wound healing is a well-orchestrated process of inflammatory response, cell proliferation, and tissue remodeling. However, this orderly and precise process is impaired in chronic wounds. A series of complicated pathogenic factors, including enhanced inflammatory response, poor cell proliferation, restrained angiogenesis, restricted collagen deposition, and infection, contribute to the failure of healing of chronic wound. The application of platelet-rich plasma (PRP) has been explored as a treatment for chronic wounds as it can balance wound microenvironment for promoting wound healing. PRP can modulate the inflammatory mediators, growth factors, and cytokines, etc. to correct abnormal biological events and disorderly molecular environment of cell migration and proliferation, and thus promote wound healing appropriately. Yet, the mechanism of PRP in promoting healing of chronic wound is still not elaborated, and the clinical application of PRP needs to be standardized as soon as possible.

Objective: To study the effect of Shengji Huayu Recipe (SHR)on the expression of MMP-3 and TIMP-1 in the skin ulcer tissue of diabetic rats., Methods: The skin ulcer model was established in diabetic mice. Different compatibility proportions of SHR [the ratio of Shengji Recipe (SJR) to Huayu Recipe (HYR) = 2:1, 1:1, and 1:2, respectively] were used to intervene. The expression of MMP-3 protein in the skin ulcer of diabetic rats was detected by Western blot method,and TIMP-1 protein was detected by immunohistochemical assay., Results: At each time point, there was no statistical difference in the blood glucose level among groups (P > 0.05). But all of them increased significantly,when compared with those of the normal wound group (P < 0.01). As for the difference between after would area treatment and before would area treatment, better effect was obtained in the SHR No. 3 group and the normal ulcer group than in the diabetic ulcer model group (P < 0.05). Results of Western blot showed that the MMP-3 protein expression was higher in the SHR No. 2 group than in the SHR No.3 group (P < 0.05). Immunohistochemical results showed that TIMP-1 protein expression was lower in the SHR No. 2 group than in the SHR No. 3 group and the diabetic ulcer model group (P < 0.05). TIMP-1 protein expression was higherin the SHR No. 3 group than in the SHR No. 2 group (P < 0.01)., Conclusion: Using SHR No.3 was conducive to the promotion of wound healing in early wound repair stage, and using SHR No. 2 might be conducive to inhibiting the formation of pathological scar.

Objective: To investigate the dynamic changes of matrix metalloproteinase-1,-9 (MMP-1, MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1 ) in the refractory diabetic dermal ulcers treated with autologous platelet-rich gel (APG)., Methods: 86 patients with nonhealing diabetic dermal ulcers were randomly assigned to two groups treated with standard procedures and APG (standard care plus topic application of autologous platelet-rich gel). The granulation tissues were collected at d0, d3, d6, d9, and d15 in patients in the APG group and at d0, d6, d15 in patients in the standard care group. The areas of ulcers were measured. The protein levels of MMP-1, MMP-9 and TIMP-1 in the tissues were determined with ELISA. The ratio of MMP-9/TIMP-1 and its relationship with the areas of ulcers were examined., Results: The areas of ulcers of patients in the APG group decreased significantely (vs. do, P < 0.05). The concentrations of MMP-1 in the granulation tissues of patients treated with APG fluctuated and reached the lowest level at d15 (vs. d6, P < 0.05). The concentrations of MMP-9 in the patients treated with AGP decreased from d3 to d15, but without statistical significance compared with d0 (P > 0.05). The concentrations of TIMP-1 in the patients treated with AGP increased from d3 and reached the peak at d6 (P < 0.05). The ratio of MMP-9/TIMP-1 at both d6 and d15 decreased significantly compared with d0 (P < 0.05) in the patients treated with AGP. The areas of ulcers in the patients with standard care decreased significantly at d15 (vs. d6, P < 0.05). The concentrations of MMP-1 reached the peak at d6 (P < 0.05) and then decreased in the patients with standard care but was still higher than the patients treated with APG (P < 0.05). The concentrations of MMP-9 decreased significantly at d15 compared with d0 in the patients treated with standard care (P < 0.05), but the change of TIMP-1 was not significant. The ratio of MMP-9/TIMP-1 in the patients with standard care decreased at d15 compared with the d0 (P < 0.05). The ratio of MMP-9/TIMP-1 was positively correlated with the areas of ulcers (r = 0.353, P < 0.05)., Conclusion: Topical application of APG might redress the proteolytic imbalance of refractory diabetic dermal ulcers by decreasing the concentration of MMPs and increasing that of TIMPs in granulation tissues. The ratio of MMP-9/TIMP-1 is a predictor of poor healing of refractory diabetic dermal ulcers.

Objective: To study the effect of repeated administration of Zhuhong ointment on renal antioxidant capability of ulcerous skin in rats, in order to further discuss the mechanism of mercury contained in Zhuhong ointment on the antioxidant capability of kidney in skin ulcer rats., Method: Eighty SD rats were randomly divided into eight groups: Zhuhong ointment A, B, C, D, E (1.219, 0.609, 0.305, 0.152, 0.76 g x kg(-1)) groups, the vaseline group, the ulcer model group and the impairment control group. The levels of NAG and RBP of toxicity for early kidney tubular injury and T-AOC, SOD, GSH-PX and GSH in kidney were determined after consecutive administration for 14 days., Result: Compared with ulcer model group, the levels of RBP in groups A, B, C and D increased, while the levels of NAG increased only in the group A. The level of T-AOC increased in groups A, B and C. The level of T-SOD increased in the group E, while it dropped down greatly in the group A. The level of GSH-PX increased in groups A, B and C. The content of GSH increased in every dose groups., Conclusion: Antioxidant capacity in rats can be increased in a reasonable dose of Zhuhong ointment, but some antioxidant activity can be notably inhibited by with the increase of dose.

Objective: To investigate the effects of Chinese herbs for replenishing qi (Yiqi) and dissolving stasis (Huayu) on substance P expression in granulation tissue of skin ulcers in rats with diabetes mellitus., Methods: One hundred male Sprague-Dawley rats with back full-thickness skin lesion were randomly divided into five groups: ulcer control group, diabetic ulcer group, Yiqi Huayu herb group, Yiqi herb group, and Huayu herb group. Except ulcer control group, the other rats were also injected streptozotocin to induce diabetes mellitus. Rats in each group were given normal saline or herbal drugs for 15 d and the wound healing rate of the rats was observed on days 8 and 15, respectively. After treatment, the expression of substance P in granulation tissues of the rats was tested with immunohistochemical method and the immunohistochemical index was calculated., Results: On the 8th and 15th days after treatment, the wound healing rate of the ulcer control group was higher than that of the diabetic ulcer group (P<0.01); the wound healing rates of all the treated groups were higher than that of the diabetic ulcer group (P<0.05); the wound healing rate of the Yiqi Huayu herb group was higher than those of the Yiqi herb group and the Huayu herb group (P<0.05). Compared with the diabetic ulcer group, the expression of substance P of the ulcer control group was increased (P<0.05); the expressions of substance P of the Yiqi Huayu herb group and the Huayu herb group were higher than that of the diabetic ulcer group (P<0.05)., Conclusion: Yiqi Huayu herbs can promote wound healing in rats with diabetes mellitus by up-regulating the expression of substance P.

Objective: To observe the effects of mild-warm moxibustion on dynamic blood flow, microvessel count (MVC)and vascular endothelial growth factor (VEGF) expression in the wound tissue of the chronic skin ulcer in rats, so as to reveal its underlying mechanism in promoting wound recovery., Methods: A total of 104 male SD rats with skin injury were randomly divided into control group (n=8), model group (n=32), TDP (far-infrared heating device) group (n=32) and moxibustion group (n=32). Chronic refractory raw surface wound model was established by muscular injection of Hydrocortisone Sodium Succinate. For rats of the TDP and moxibustion groups, TDP irridiation and mild-warm moxibustion were applied to the raw surface, bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) for 15 min, once daily for 3, 7 and 14 days respectively. The healing rate and the healing time of raw surface of the wound were observed. The blood flow of the raw surface of the wound tissue was measured by laser Doppler flowmeter and the MVC in granulation tissue of chronic skin ulcer was counted under light microscope. VEGF expression was detected by immunohistochemistry., Results: In comparison with the control group, the healing rate of the wound raw surface was significantly lower and the healing time was prolonged in the model group (P < 0.01). Compared with the model group, the healing rates on day 3, 7, 10 and 14 were significantly higher and the healing time was strikingly faster in both TDP and moxibustion groups (P < 0.01, P < 0.05), and the effects of the moxibustion group in increasing the healing rate and shortening the healing time were significantly better than those of TDP group (P < 0.01). In comparison with the model group, the blood flow volume, MVC and VEGF expression levels on day 3 and 7 were upregulated significantly in both TDP and moxibustion groups (P < 0.01, P < 0.05); while the blood flow volume, MVC and VEGF expression level in the moxibustion group and the blood flow volume and VEGF expression level in the TDP group downregulated considerably on day 14 (P < 0.01). No significant difference was found between the TDP and moxibustion groups in the MVC on day 14 after the treatment (P > 0.05)., Conclusion: Mild-warm moxibustion can promote wound healing, which is closely with its effects in increasing blood flow and MVC, and upregulating VEGF expression in the wound granulation tissue of the chronic skin ulcer.

Background and Objective: Aloe has preventive effects on some chemotherapy-induced extravasation injuries. This study was to investigate the effect and mechanism of aloe gel on doxorubicin-induced extravasation injury., Methods: Sprague-Dawley (SD) rats were used to establish the extravasation injury model induced by doxorubicin. Thirty SD rats were randomly divided into three groups: control group, aloe gel group (1 g/L) and 50% magnesium sulfate group. The area of extravasation was measured and the degree of injury was observed. The injured tissues were resected from two randomly selected rats in each group on the 1st, 4th, 7th, 11th, and 18th day after treatments. Pathological morphology of the resected tissues was observed under an optical microscope after hematoxylin and eosin (HE) staining. The exosmosis skin and subcutaneous tissues of rats were resected five days after treatments. Then the wounds were interruptedly sutured. When sutures were removed on the 7th day after operation, the condition of primary wound healing and the healing time were recorded. Expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) in the exosmosis skin and subcutaneous tissues were detected by immunohistochemistry., Results: The area and the degree of extravasation injury were smaller and less severe in the aloe gel and magnesium sulfate groups than in the control group (P<0.01). The rates of primary wound healing were significantly higher in the aloe gel (60.0%) and magnesium sulfate (66.7%) groups than in the control group (20.0%); while the healing time was significantly shorter in the aloe gel (9.6+/-1.64 d) and magnesium sulfate (9.33+/-1.40 d) groups than in the control group (12.13+/-2.06 d) (both P<0.01). Moreover, the expression levels of VEGF and EGFR were higher in the aloe gel group than in the control group., Conclusion: The preventive and therapeutic effects of aloe gel on doxorubicin-induced extravasation injury are satisfactory, which may be in relation to the up-regulation of VEGF and EGFR.

Objective: To explore the effects of reinforcing qi and nourishing blood recipes on vascular endothelial growth factor (VEGF) and the microvessel count (MVC)in granulation tissue in rats with chronic skin ulcers., Methods: A total of 90 male SD rats with back full-thickness skin lesion were used in this study. Except for the normal control group, the rats were injected with hydrocortisone for inducing chronic skin ulcers, and were randomly divided into untreated group, Sijunzi Decoction group, Siwu Decoction group and Bazhen Decoction group. The time of wound healing was observed; immunohistochemical method and image analytical method were used to test the expressions of VEGF and MVC in granulation tissue of dermal chronic ulcers in the rats., Results: In the untreated group, the time of wound healing was significantly longer than that in the normal group (P<0.01). The time of wound healing was obviously shortened in the three treated groups as compared with the untreated group (P<0.01, P<0.05). In the Sijunzi Decoction group, the time of wound healing was obviously shorter than those in Siwu Decoction group and Bazhen Decoction group (P<0.05, P<0.01). In the untreated group, the expression of VEGF and MVC were significantly lower than those in the normal control group (P<0.01). The expression of VEGF and MVC were obviously increased in the three treated groups as compared with those in the untreated group (P<0.01, P<0.05). In the Sijunzi Decoction group, the expression of VEGF and MVC were obviously lower than those in Siwu Decoction group and Bazhen Decoction group (P<0.05, P<0.01)., Conclusion: Reinforcing qi and nourishing blood recipes can promote wound healing in rats through up-regulating the expression of VEGF in granulation tissue of dermal chronic ulcers, and inducing angiogenesis.

Objective: To study the influence of maggot secretion on expression of bFGF and connective tissue growth factor (CTGF) in ulcer tissue of diabetes mellitus (DM) rats and its antibacterial function., Methods: There were 40 3-month-old SD male rats (weighing 300-350 g) which were randomly divided into 2 groups: control group and experimental maggot secretion group. The model of ulcer wound of DM rats was made. The ulcer wound of DM rats in maggot secretion group spread maggot secretions, but no secretion on ulcer wound was found in control group. The morphological and tissue changes of ulcer wound were observed at different times, and the conditions of bacterial infection on ulcer wound in the two groups were checked. Tissue slices were prepared on 7, 14 and 21 days, respectively; immunohistological detection of bFGF and CTFG in ulcer wound of the two groups was done; and the cell number of positive expression of bFGF and CTFG was counted., Results: It was found that the healing of ulcer was dominant in experimental group; the wound was clean; the tissue regenerated and no Staphylococcus aureus infection was seen. Bad healing was obtained in control group; tissue necrosis was found and the rate of Staphylococcus aureus infection was 60%. Positive expression cell number of bFGF in ulcer wound was detected on 7 and 14 days after operation with 23.76 +/- 3.34 and 52.76 +/- 4.84 in experimental group, and 18.88 +/- 2.16 and 46.04 +/- 4.00 in control group. Positive expression cell number of CTGF in ulcer wound was detected on 7 and 14 days after operation with 18.76 +/- 3.24 and 46.52 +/- 4.07 in experimental group, and 12.52 +/- 3.03 and 40.52 +/- 3.96 in control group. There was significant difference between positive expressions of bFGF and CTFG in the two groups (P < 0.05)., Conclusion: The maggot secretion can elevate the expressions of bFGF and CTFG in ulcers, promote healing and prevent bacterial infection.

Objective: To study the effects of resolving stagnation and promoting granulation therapy on expressions of Bax and Bcl-2 in granulation tissue of diabetic rats during wound healing., Methods: Seventy-two male SD diabetic rats with full-thickness skin lesion were randomly divided into 3 groups: SJHYR 1-treated group, SJHYR 2-treated group and normal saline (NS) control group. SJHYR 1 was prepared with Shengji Recipe (SJR, a compound traditional Chinese herbal medicine for promoting granulation) and Huayu Recipe (HYR, a compound traditional Chinese herbal medicine for resolving stagnation) at a ratio of 1:2, while SJHYR 2 was prepared with SJR and HYR at a ratio of 1:1. Immunohistochemical method was used to assess Bax and Bcl-2 protein levels in granulation tissue., Results: SJHYR 1 could accelerate wound healing as compared with SJHYR 2 and NS (P<0.05). On the third day in experiment, Bax and Bcl-2 proteins were not found in any groups, but on the seventh and eleventh day in experiment, Bax and Bcl-2 proteins in SJHYR 1-treated group were much higher than those in the other two groups (P<0.05)., Conclusion: SJR and HYR in different ratios may all have a role in regulating Bax and Bcl-2 expression in granulation tissue of diabetic rats during wound healing.

Objective: To explore the effects of Yiqi Huayu Recipe, a compound traditional Chinese herbal medicine for replenishing qi and resolving stagnation, on transforming growth factor-beta1(TGF-beta1) of skin ulcers in rats with diabetes., Methods: Sixty rats were randomly divided into six groups, Yiqi Huayu Recipe-treated group, Yiqi (replenishing qi) Recipe-treated group, Huayu (resolving stagnation) Recipe-treated group, basic fibroblast growth factor (bFGF) treated group, untreated group and normal control group. Diabetes was induced by peritoneal injection of streptozotocin and skin ulcers were made by surgery method in rats except for the normal control group. Then the rats were administered with different drugs respectively, and the expression of TGF-beta1 in granulation tissue of the skin ulcers was detected with the methods of Western blotting, image analysis and immunohistochemistry., Results: The level of TGF-beta1 expression in the untreated group was lower than that in the normal control group (P<0.01); and the level of TGF-beta1 expression in the drug-treated groups was higher than that in the untreated group (P<0.01); and the TGF-beta1 expression in the Yiqi Huayu Recipe-treated group was higher than that in the Yiqi Recipe-treated group, Huayu Recipe-treated group and bFGF-treated group (P<0.05)., Conclusion: The replenishing qi and resolving stagnation therapy can control the secretion of TGF-beta1 of the wound in the process of wound healing in the levels of gene and molecule.

Objective: To explore the effects of Chinese herbs for replenishing qi and resolving stagnation on hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in granulation tissue of skin ulcers in rats with syndrome of blood stasis and qi deficiency., Methods: Diabetic rats with back full-thickness skin lesion and syndrome of blood stasis and qi deficiency were divided in to five groups: untreated group, basic fibroblast growth factor (bFGF)-treated group, Yiqi Huayu Recipe (a recipe for replenishing qi and resolving stagnation)-treated group, Yiqi Recipe (a recipe for replenishing qi)-treated group and Huayu Recipe (a recipe for resolving stagnation)-treated group, and another eight normal rats served as normal control group. Immunohistochemical method and image analysis were used to test the expressions of HIF-1alpha and VEGF in granulation tissue of skin ulcers in rats with diabetes., Results: In the untreated group, the expression of HIF-1alpha was significantly increased and the expression of VEGF was significantly decreased as compared with those in the normal control group (P<0.01). The expression of HIF-1alpha was obviously lower (P<0.01) and the expression of VEGF was significantly higher in the four drug treated groups as compared with those in the untreated group. In the Yiqi Huayu Recipe-treated group, the expression of HIF-1alpha was obviously lower than those in the bFGF-treated, Yiqi Recipe-treated and Huayu Recipe-treated groups (P<0.05 or P<0.01), and the expression of VEGF was significantly higher than that in the bFGF-treated group (P<0.01)., Conclusion: Chinese herbs for replenishing qi and resolving stagnation can promote the wound healing in rats through reducing the expression of HIF-1alpha, accelerating the expression of VEGF in granulation tissue of skin ulcers in rats with diabetes and ameliorating the status of ischemia and hypoxia.

Objective: To investigate the dynamic expression of platelet-derived growth factor-A (PDGF-A) and its receptor alpha (PDGFR-alpha) in different acute radiation-induced skin ulcers, and to explore the underlying mechanism involved in retarded healing of the ulcer., Methods: The model of acute radiation-induced skin ulcers in rats was replicated with 50 Gy 60Co gamma rays to the skin (radiation group, R, n = 55), rats with full - thickness skin excision wounds as control group (T, n = 55), and 5 normal rats to serve as normal control (NC) group. The expression of PDGF-A and PDGFR-alpha protein and PDGF-A mRNA was respectively assessed by means of histochemistry and in situ RT-PCR., Results: No PDGF-A expression was identified in the rat skin in NC group. The expression of PDGF-A and PDGFR were reduced in R group during inflammatory responsive and granulation formation periods (14 - 28 days after radiation, the IA value of PDGF-A varied from 14.0 +/- 1.2 to 20.3 +/- 1.2 compared with that in T group in which the IA value of PDGF-A at the same period (3 - 9 days after injury) varied from 20.0 +/- 1.6 to 28.3 +/- 1.0, and reduced gradually during scar formation period (55 days after radiation)., Conclusion: The reduction of PDGF-A and PDGFR-expression may be partially involved in the mechanism of retarded healing of acute radiation-induced skin ulcers.

Objective: To investigate the expression and location of epidermal growth factor (EGF) and its receptor (EGFR) in dermal chronic ulcers and normal skin in order to explore their influence on ulcer formation., Methods: The expression intensity and distribution of EGF and EGFR were detected with pathological method and immunohistochemistry method in 8 cases of dermal ulcers, 8 cases of edge of ulcer and 8 cases of normal skin., Results: The Positive signals of EGF could be found in epidermal cells, endothelial cells and some fibroblasts; EGFR was principally located in the cytoplasm and cellular membrane of these cells mentioned above in normal skin. From normal skin, edge of ulcer to ulcerative tissues, the protein contents of EGF and EGFR were decreased progressively. In ulcerative tissues, EGF was mostly distributed in monocytes and macrophages while EGFR was chiefly sited in monocytes. When compared with normal skins, the protein expression of EGF and EGFR was notably reduced in ulcerative tissues (both P<0.01). The positive cellular ratios of two proteins were reduced to (7.1+/-5.2) % and (8.8+/-5.5) % of those in normal skin respectively (all P<0.01)., Conclusion: The formation of dermal chronic ulcers is closely associated with the reduction of EGF and EGFR protein expression which may lead to binding obstruction between EGF and its receptor.

Objective: To investigate the characteristics of PCNA expression in hypertrophic scars and chronic ulcers and to discuss its relation to their formation., Methods: The expressive quantity and sites of PCNA were detected with the immunohistochemical SP method., Results: PCNA was expressed in all samples. The expressive quantity in hypertrophic scars was higher than chronic ulcers(P < 0.01). The expressive sites of all samples were in the nucleus of fibroblasts and capillary endothelial cells., Conclusions: The expressive quantity of PCNA was more in hypertrophic scars and less n chronic ulcers. The quantitative difference of expression between hypertrophic scars and chronic ulcers may be correlated to their formation.

Objective: To localize the distribution of basic fibroblast growth factor (bFGF) and transforming growth factor-beta(TGF-beta) in tissues from dermal chronic ulcer and hypertrophic scar and to explore their effects on tissue repair., Methods: Twenty-one cases were detected to localize the distribution of bFGF and TGF-beta, among them, there were 8 cases with dermal chronic ulcers, 8 cases with hypertrophic scars, and 5 cases of normal skin., Results: Positive signal of bFGF and TGF-beta could be found in normal skin, mainly in the keratinocytes. In dermal chronic ulcers, positive signal of bFGF and TGF-beta could be found in granulation tissues. bFGF was localized mainly in fibroblasts cells and endothelial cells and TGF-beta mainly in inflammatory cells. In hypertrophic scar, the localization and signal density of bFGF was similar with those in granulation tissues, but the staining of TGF-beta was negative., Conclusion: The different distribution of bFGF and TGF-beta in dermal chronic ulcer and hypertrophic scar may be the reason of different results of tissue repair. The pathogenesis of wound healing delay in a condition of high concentration of growth factors may come from the binding disorder of growth factors and their receptors. bFGF may be involved in all process of formation of hypertrophic scar, but TGF-beta may only play roles in the early stage.

Objective: To determine the characteristics and regularity of fibronectin mRNA expression in diabetic ulcers, and to investigate the relationship between the changes of fibronectin mRNA expression and pathogenesis of diabetic ulcer., Methods: Biopsies were removed from the margins of diabetic foot ulcers, included surrounding skin as experimental group, and the biopsies from normal skin of the same patients as control group. The mRNA expression of fibronectin was measured by quantitative RT-PCR technique., Results: The mRNA expression of fibronectin could be detected in both normal skin and diabetic foot ulcers, but the level of expression in diabetic ulcers was lower than that of normal skin., Conclusion: The level of mRNA expression of fibronectin in diabetic ulcers is decreased, which suggest that the down-regulation of transcription may be one of the mechanisms of chronic impaired ulcers.