Arginine Vasopressin-Aquaporin-2 Pathway-Mediated Dehydration Effects of Electroacupuncture in Guinea Pig Model of AVP-Induced Eendolymphatic Hydrops.

OBJECTIVE: To investigate the effects of electroacupuncture (EA) on endolymphatic hydrops (EH) and the regulation of arginine vasopressin (AVP)-aquaporin-2 (AQP2) pathway in guinea pigs. METHODS: EH was induced in male guinea pigs by an intraperitoneal injection of AVP. For the treatment, EA was delivered to Baihui (GV 20) and Tinggong (SI 19) acupoints, once per day for 10 consecutive days. In histomorphological studies, cochlear hydrops degree was evaluated by hematoxylin-eosin (HE) staining, and then the ratio of scala media (SM) area to SM + scala vestibuli (SV) area (R value) was calculated. In mechanical studies, a comparison of plasma AVP (p-AVP) concentrations, cyclic adenosine monophosphate (cAMP) levels, vasopressin type 2 receptor (V2R) and AQP2 mRNA expressions in the cochlea were compared among groups. RESULTS: EA significantly reduced cochlear hydrops in guinea pigs (P=0.001). EA significantly attenuated the AVPinduced up-regulation of p-AVP concentrations (P=0.006), cochlear cAMP levels (P=0.003) and AQP2 mRNA expression (P=0.016), and up-regulated the expression of V2R mRNA (P=0.004) in the cochlea. CONCLUSIONS: The dehydrating effect of EA might be associated with its inhibition of AVP-AQP2 pathway activation.

[Analgesic Effect and Mechanism of Electroacupuncture on Rats with Chronic Inflammatory Pain].

OBJECTIVE: To observe analgesic effect of electroacupuncture ( EA) on rats with chronic inflammatory pain and its regulatory mechanism on ispilateral dorsal root ganglion (DRG) and spinal dorsal horn (SDH) Mas-related G protein-coupled C receptor (MrgprC). METHODS: Totally 40 healthy male SD rats were divided into 4 groups according to random number table, i.e., the normal (N) group, the model (M) group, the acupuncture (Acu) group, the EA group, 10 rats in each group. The model of chronic inflammatory pain was established by subcutaneous injecting 0. 1 mL complete Freund's adjuvant (CFA) into right hind paw. Paw withdrawal thresholds (PWTs) were measured before modeling, at day 1, 3, 5, 7, and after CFA injection, respectively. Expression levels of MrgprC in ispilateral DRG and SDH were detected by Western blot. The content of bovine adrenal medulla 22 (BAM22) in SDH was detected by immunohistochemical assay. RESULTS: Compared with N group at each time point, PWTs significantly decreased in M group (P <0. 01). Compared with M group, PWTs significantly increased at day 5 of EA and after EA in EA group (P < 0.05, P < 0.01). Compared with Acu group at each time point, post-EA PWTs significantly increased in the EA group (P < 0.05). Compared with N group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in M group (P < 0.01). Compared with M group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in the EA group (P < 0.05). CONCLUSION: EA had favorable analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism might be possibly associated with up-regulating MrgprC expression in ispilateral DRG and BAM22 content in ispilateral SDH.

[Study on rules of acupoints selection for vertigo in ancient acupuncture].

The characteristics of selection and matching acupoint and application rules of meridians in ancient acupuncture literature for vertigo were explored. The data were collected from literature regarding acupuncture for vertigo from the pre-Qin period to Qing dynasty and then database was established. Frequency statistics method and comparison of support degree were applied to analyze and explore application rules of acupoints and meridians, while association rules in data mining was used to extract combinations of acupoints. As a result, for treatment of vertigo, according to frequency of use and support degree, generally the most selected acupoints, in turn, were Fengchi (GB 20), Shangxing (GV 23), Yanggu (SI 5), Jiexi (ST 41), Zulinqi (GB 41) and Shenting (GV 24), etc.; the most methods for matching acupoint were combination of adjacent acupoints, combination of same-meridian acupoints, combination of the superior-inferior acupoints, combination of yang-meridian and yang-meridian acupoints and combination of child-mother meridians acupoints; the most selected meridians were bladder meridian of foot-taiyang, gallbladder meridian of foot-shaoyang and governor vessel. Compared between the ancient and modern literature, it was found out that the ancient and modern clinic has same points in selection of acupoint-meridian and matching acupoints methods. However, the use of Yanggu (SI 5), Jiexi (ST 41) and Feiyang (BL 58) as well as combination of child-mother meridians acupoints were less seen in modern clinic, which could provide new reference.

[Clinical application of low-molecular-weight heparin (Fraxiparine) in rescuing venous crisis of island skin flap].

OBJECTIVE: To evaluate the clinical efficacy of low molecular weight heparin (Fraxiparine) in rescuing venous crisis of island skin flap. METHODS: Of the 73 patients with venous crisis of island skin flap, 47 received subcutaneous injection of low-molecular-weight heparin (group I) and 26 were treated with phlebotomy, local compression and topical application of unfractionated heparin solution gauze (group II). RESULTS: The flap survival ratio was (88.46∓8.64)% in group I and (38.37∓6.53)% in group II (P<0.001). At 0, 2, and 4 h after injection of low-molecular-weight heparin, the activated partial thromboplastin time (APTT) was obviously delayed (24.28∓6.71, 41.35∓7.64 and 32.34∓6.35, respectively, P<0.01), FXa:C level was significantly decreased (152.4∓30.7, 65.8∓24.4 and 83.4∓18.4, respectively, P<0.01), while FIIa:C level underwent no obvious alterations (155.70∓31.61, 143.20∓24.75, and 143.4∓23.35, respectively, P=NS). CONCLUSION: Fraxiparine has good antithrombotic efficacy in rescuing venous crisis of island skin flap without adverse effect on systemic coagulation.

Normal skin and hypertrophic scar fibroblasts differentially regulate collagen and fibronectin expression as well as mitochondrial membrane potential in response to basic fibroblast growth factor

Basic fibroblast growth factor (bFGF) regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin and from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-inducedeffects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts from tissue biopsies of patients who underwent plastic surgery for repairing hypertrophic scars. The fibroblasts were then treated with different concentrations of bFGF (ranging from 0.1 to 1000 ng/mL). The growth of hypertrophic scar fibroblasts became slower with selective inhibition of type I collagen production after exposure to bFGF. However, type III collagen expression was affected in both normal and hypertrophic scar fibroblasts. Moreover, fibronectin expression in the normal fibroblasts was up-regulated after bFGF treatment. bFGF (1000 ng/mL) also induced mitochondrial depolarization in hypertrophic scar fibroblasts (P < 0.01). The cellular ATP level decreased in hypertrophic scar fibroblasts (P < 0.05), while it increased in the normal fibroblasts following treatment with bFGF (P < 0.01). These data suggest that bFGF has differential effects and mechanisms on fibroblasts of the normal skin and hypertrophic scars, indicating that bFGF may play a role in the early phase of skin wound healing and post-burn scar formation.

Normal skin and hypertrophic scar fibroblasts differentially regulate collagen and fibronectin expression as well as mitochondrial membrane potential in response to basic fibroblast growth factor.

Basic fibroblast growth factor (bFGF) regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin and from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-induced effects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts from tissue biopsies of patients who underwent plastic surgery for repairing hypertrophic scars. The fibroblasts were then treated with different concentrations of bFGF (ranging from 0.1 to 1000 ng/mL). The growth of hypertrophic scar fibroblasts became slower with selective inhibition of type I collagen production after exposure to bFGF. However, type III collagen expression was affected in both normal and hypertrophic scar fibroblasts. Moreover, fibronectin expression in the normal fibroblasts was up-regulated after bFGF treatment. bFGF (1000 ng/mL) also induced mitochondrial depolarization in hypertrophic scar fibroblasts (P < 0.01). The cellular ATP level decreased in hypertrophic scar fibroblasts (P < 0.05), while it increased in the normal fibroblasts following treatment with bFGF (P < 0.01). These data suggest that bFGF has differential effects and mechanisms on fibroblasts of the normal skin and hypertrophic scars, indicating that bFGF may play a role in the early phase of skin wound healing and post-burn scar formation.

[Establishment of a rat model of bone marrow mesenchymal stem cell transplantation for repairing full-thickness skin defect].

OBJECTIVE: To establish a rat model of full-thickness skin defect to receive bone marrow mesenchymal stem cell transplantation for wound repair. METHODS: A full-thickness skin defect measuring 4 cmx4 cm in 36 F344 rats, which were divided into 3 groups with the wound covered with alloskin graft, acellular dermal matrix, or petrolatum gauze. In vitro cultured BMSCs in the 5th passage were transplanted into the skin defect, and the time of wound dressing dissociation and number of transplanted Brdu-positive cells in the wound were observed 14 days later. RESULTS: The alloskin graft resulted in significantly longer time before dressing dissociation, with greater number of Brdu-positive cells in the wound than the other two wound dressings (P<0.001). The acellular dermal matrix showed better effect than petrolatum gauze in terms of the dressing dissociation time and the viable transplanted cell number in the wound. CONCLUSION: Alloskin graft can be ideal for covering the wound surface to protect the transplanted BMSCs in rats.

Basic fibroblast growth factor (bFGF) alleviates the scar of the rabbit ear model in wound healing.

Studies suggest a possible antiscarring effect of basic fibroblast growth factor (bFGF) during wound healing. However, little is known about the precise pathological mechanisms of bFGF. In particular, there is only limited information available about the mechanism of exogenous administration of bFGF to scar formation. To investigate the effect of bFGF on the hypertrophic scar in the rabbit ear model and to clarify the mechanisms of bFGF on treatment for scar in wound healing, the rabbit ear model of wound healing was created and treated topically with bFGF once daily for 3 months; then we examined the changes of macroscopic and histopathological characteristics of scars and the expression of collagen and collagenase-1 (matrix metalloproteinase-1). The results of macroscopic and histologic characteristics revealed a significant difference between scars treated with bFGF and control scars. The expression of collagen in the scars treated with bFGF was decreased, as compared with the scars treated with saline. Further study revealed that bFGF could remarkably enhance expression of matrix metalloproteinase-1. bFGF could improve the quality of wound healing and remarkably alleviate the scar in the rabbit ear model in wound healing, which suggests that bFGF exerted a net negative effecton scar formation in wound healing. The evidence should contribute to a better understanding of the biological activities of bFGF during hypertrophic scar formation.

[Establishing a parabiosis animal model of two-way paradigm].

OBJECTIVE: To establish a parabiosis model between allogenic conspecific adult mice to study two-way paradigm. METHODS: Fifty-four female Balb/c mice and 54 male C57BL/6 mice were paired and equally divided into 3 groups, namely group 1 with normal saline (NS) injection, group 2 with injections of spleen cells and cyclophosphamide (CP), and group 3 injected with spleen cells, CP, and cyclosporin A (CsA). The treatments were performed by injecting the spleen cells from one of the mice in a pair into the other via tail vein and vise versa, and two days after the operation, CP (150 mg/kg) was injected intraperitoneally. Intraperitoneal CsA (30 mg/kg daily) injection was given starting from 2 days before till 7 days after the operation. Twelve of the 18 pairs of parabiosis mice in each group were separated after 1 week, and part of the skin were transplanted to each other. The maintenance of parabiosis was observed in the other 6 pairs of parabiosis mice were observed. Mixed lymphocyte reaction (MLR) and delayed-type hypersensitivity (DTH) were observed and studied in the separated mice. RESULT: The duration of parabiosis maintenance and skin survival of the group 3 was significantly longer than those in the other two groups, and group 3 showed suppressed MLR and DTH. CONCLUSION: With the application of immunosuppressants, we have successfully established the two-way paradigm model in mice.

[Application of laminar air flow techniques in burn treatment].

OBJECTIVE: To evaluate the value of laminar flow in the treatment of burns. METHODS: The air in the laminar flow chamber and the wound tissues of the patients were sampled for bacterial detection. The number and stains of bacterial colony from different classes of laminar air flow chambers at different time points were inspected and compared. RESULTS: The bacterial number was 0 in the laminar flow chamber of 1000 grade, which was obviously different from that in the public area. The mortality was obviously decreased in the laminar air flow chamber with shorter treatment time and hospitalization. No wound infection occurred and the wounds healed smoothly in all these patients. CONCLUSION: The application of laminar air flow can be helpful for the treatment of severe burns.

[Vascular endothelial growth factor gene transfection and survival of the random skin flap in rats].

OBJECTIVE: To investigate the effects of vascular endothelial growth factor (VEGF) gene transfection on survival of the random skin flap in rats. METHOD: Thirty SD rats were randomized equally into 3 groups: pcDNA3-VEGF165, pcDNA3 and control groups, with the former two groups transfected via liposome with pcDNA3-VEGF165 and pcDNA3 respectively 48 h before and during the operation. Ischemic random skin flaps ( 1 cmx7 cm) were constructed from the rats. Seven days later, the amount of viable tissue within the flap was measured by planimetry. After the animals were killed, and specimens from the random skin flaps were harvested for immunohistologic evidence of VEGF protein expression and for HE staining to examine the microvascular growth. RESULTS: The results of tissue survival planimetry of the skin flap of pcDNA3-VEGF165, pcDNA3 and control groups were 48.46% +/-3.35%, 30.20%+/-2.16%, and 31.35% +/-1.99%, which were highest in the VEGF- transfected group (P<0.05), in which immunohistochemical staining revealed increased deposition of VEGF in comparison with the other control groups P<0.05 . The VEGF group had also higher average vessel number as compared with the vector and control group (107.72+/-9.42 vs 91.35+/-7.28 and 89.85+/-7.66, P<0.05), and smaller average vessel lumen diameter (25.76+/-3.23 microm vs 32.12+/-1.58 microm and 33.49+/-2.29 microm, P<0.05). CONCLUSION: pcDNA3-VEGF165 transfection may enhance the survival of the ischemic skin flaps and achieve VEGF expression in the flaps in rats.

[Change of protein catabolism and immunocompetence in severely burned patients after operations and the therapeutic effect of recombinant human growth hormone].

OBJECTIVE: To investigate the changes in protein catabolism and immunocompetence in severely burned patients after operations and observe the therapeutic effect of recombinant human growth hormone (rhGH) in these patients. METHODS: The changes in plasma protein concentration and immune function and the clinical results of 38 severely burned patients after three operations were retrospectively analyzed. These patients were assigned to GroupI(control, n=20) and GroupII (rhGH treatment, n=18), and matched for age, total body surface area (TBSA), surgical approaches and times, perioperative bleeding volume and nutrition support etc. The patients in groupII received intramuscular injection with 0.2-0.4 U/d x kg x b.w. rhGH after the first operation, till the healing of the wounds. RESULTS: The concentration of retina-binding protein, transferrin, prealbumin, albumin, and IgG, the number of CD4 and CD8 cells, as well as the ratio of CD4/CD8 cells prior to the operation were below the normal levels in the two groups of patients. All these indexes in GroupII recovered to a great extent after three operations and were significantly higher than those in GroupI(P<0.05). CONCLUSION: Nutritious state and immunocompetence of patients with severe burns are severely compromised after operations, and nutrition support and rhGH treatment may produce rapid improvement of the patients' conditions.