OBJECTIVETo establish a cell line for stable expression of human beta-defensin 3 (hBD3).

METHODSFull length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofectamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated.

RESULTSCOS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity.

CONCLUSIONWe successfully established a hBD3-expressing cell line.

Animals ; COS Cells ; Cercopithecus aethiops ; Escherichia coli ; drug effects ; Genetic Vectors ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Staphylococcus aureus ; drug effects ; Transfection ; beta-Defensins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo establish a long-term in vitro culture of the fibroblasts obtained from burn wounds.

METHODSSkin samples were harvested from normal volunteers and the deep partial thickness burn wound in burn patients on the 5th, 10th, 21st, 28th and 35th postburn days (PBDs). The non-dermal tissue was removed from the samples and primed by chlorhexidine solution in concentration of 2.5 g/L. The skin sample was then digested by trypsin-EDTA in concentration of 1.25 g/L and was centrifuged before the cells were harvested and cultured. When the cells grew nearly to form sheet, multiple passage culture, freezing storage and revivification were carried out with routine methods. The cell morphology was continuously observed during the culture. And the cell doubling time was calculated.

RESULTSThe wound-origin fibroblasts exhibited higher purity and better activity. The cellular growth features and gross morphology kept stable during primary and secondary culture, and during freezing storage and after revivification. The cells kept their activity above 80% of their original after many times of revivification.

CONCLUSIONThe establishment of the in vitro culture of fibroblasts from burn wounds might be useful in the exploration of the pathogenesis and therapeutic measures of scars.

Burns ; metabolism ; pathology ; Cell Culture Techniques ; methods ; Cell Division ; Cell Survival ; Cells, Cultured ; Cryopreservation ; Factor VIII ; analysis ; Fibroblasts ; chemistry ; cytology ; Humans ; Immunohistochemistry ; Time Factors

OBJECTIVETo investigate the clinical usage of percutanious retrograde acetabular anterior horn screw fixation.

METHODSFrom June 2006 to November 2011,27 patients with anterior horn accetabular fractures were treated with percutaneous retrograde screw. There were 20 males and 7 females with an average age of 35 years (ranged,20 to 61). According to Tile classification,type A was in 5 cases,type B was in 18 cases and type C was in 4 cases. The intra-operative blood loss volume, fractured reduction and screw location were observed.

RESULTSPostoperative X-ray and CT scan showed good reduction. According to the standard of Matta, 10 cases got excellent results, 15 good and 2 poor. All fractures healed and no complications such as neurovascular injuries and femoral head necrosis were found.

CONCLUSIONPercutanious retrograde acetabular anterior horn screw technique have advantages of little trauma, less blood loss volume, reliable fixation, which can be effectively used in clinic.

Acetabulum ; diagnostic imaging ; injuries ; surgery ; Adult ; Bone Screws ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Radiography

To explore new measures for functional reconstruction of multiple severe deformities as a result of extensive deep burn (total burn surface area > or = 90% TBSA, including deep burn > or = 70%TBSA) in late stage. Twelve severe burn patients with above-mentioned deformities were hospitalized in our ward during 1960--2005, the scars resulted from burns were distributed from head to foot with 173 deformities, including 27 scar ulcers. All patients lacked of self-care ability, among them some could not stand. Due to inadequate skin source, deformities were corrected by skin from matured scars expanded with subcutaneous balloon at late postburn stage. Following our former clinical experience, anatomic investigation and experimental research, we chose the following methods to correct deformities and restore functions: application of split-thickness scar skin after expansion (88 wounds); use of scar skin flap/scar-Achilles tendon flaps (59 wounds); combination of thin split-thickness skin grafts from scar and allogeneic acellular dermal matrix (composite skin, 40 wounds). All grafts survived, the appearance and function were improved obviously without complications. Follow-up 1-40 years, all patients could take care themselves with satisfactory function and appearance, and among them 8 patients returned to work (one had worked for 40 years), 2 patients married and had children. The above-mentioned measures are safe, reliable and effective for functional reconstruction of deformities.
Adult ; Burns ; complications ; surgery ; Cicatrix ; etiology ; surgery ; Contracture ; etiology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Recovery of Function ; Skin Transplantation ; Skin, Artificial ; Surgical Flaps ; Wound Healing

OBJECTIVETo investigate the optimal operation method for the management of various chronic wounds in legs and feet.

METHODSFifty-one chronic wounds were evaluated according to infection, inflammatory response, and distribution in different areas of the leg and foot. Preoperative treatment was given accordingly, then transposition of skin flap, skin grafting, or amputation was performed. The healing rate after single session operation and average hospitalization were statistically analyzed.

RESULTSThe wound healing rate after single session operation was 86. 3% , the average hospital stay was (17. 8 +/- 2. 1) days, and the appearance and function of the leg and foot after operation was satisfactory.

CONCLUSIONThe appropriate preoperative treatment and operation method conforming to the wound location and evaluation are of vital importance in the management of chronic wounds in the leg and foot. Operation is one of the most effective ways to repair chronic wounds in the leg and foot, and it can shorten the wound healing process and restore the function.

Adult ; Chronic Disease ; Foot Ulcer ; pathology ; surgery ; Humans ; Leg Ulcer ; pathology ; surgery ; Longevity ; Male ; Surgical Flaps ; Wound Healing

OBJECTIVETo investigate the change in neuropeptide substance P (SP) in the burn wound skin in scalded rats.

METHODSThe SP distribution density in the rat skin samples from the burn wound, peri-wound area and remote area of scalded rats were determined by immunohistochemistry technique. The mRNA expression of SP was observed by in situ hybridization.

RESULTSThe distribution density of SP in the skin samples from the above three areas decreased obviously since 15 mins postburn, dropped to the lowest level during 6 - 12 postburn hou (PBH), and then recovered thereafter. Comparatively, the recovery in peri-wound area occurred earlier. In addition, there was emigration of macrophage-like large cells with positive SP immunoreactivity from local blood vessels in the dermal layer of the burn wound and peri-wound areas. The large cells were closely related to the local SP-containing nerve fibres at 12 PBHs. Intensified staining of the cells was identified at 24 PBHs with the disintegration of the cells into pieces with release of large amounts of positive SP immunoreactive granules, followed by disappearance of the cells at 48 PBHs. It was indicated by in situ hybridization that the preprotachykinin A (PPTA) mRNA encoding and synthesizing SP was expressed in various cells at 6 PBHs.

CONCLUSIONSP might be synthesized and released from cutaneous nerve ending or from local inflammatory cells after skin burn injury.

Animals ; Burns ; metabolism ; pathology ; In Situ Hybridization ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; Substance P ; metabolism ; Wound Healing ; physiology

OBJECTIVETo investigate the feasibility of in vitro proliferation of rat Leydig cells by modifying the cell culture system.

METHODSLeydig cells were isolated from three-week-old rats by a procedure combining collagenase dispersion, stainless steel mesh infiltration and differential adhesion. The isolated cells were cultured in DMEM/F12 and modified media for stem cell proliferation, and the proliferation of the cultured cells was evaluated by cell counting and MTP test. The expression of 3beta-HSD in the cultured cells was detected by immunohistochemistry and flow cytometry, and testosterone productivity in the isolated Leydig cells with or without hCG stimulation was determined at 2 hours and 4 days after cell isolation.

RESULTSThe Leydig cells cultured in the modified media proliferated actively, with a doubling time of (2.26 +/- .31) days, as compared with (16.32 +/- 2.14) days for those cultured in the traditional media (P <0.05). The 3beta-HSD positive rate in the cultured cells was (554.3 +/- 7.1)% after 2 hours and (93.6 +/- 4.6)% after 4 days of culture. All the proliferated cells exhibited testosterone productivity, and their testosterone secretion was significantly upregulated by hCG stimulation (P <0.05).

CONCLUSIONLeydig cells isolated by differential adhesion proliferate actively in the modified culture media.

Animals ; Cell Count ; Cell Culture Techniques ; Cell Proliferation ; Cells, Cultured ; Leydig Cells ; cytology ; Male ; Rats ; Rats, Wistar ; Testosterone ; secretion

OBJECTIVETo study the risk factors of acute renal insufficiency (ARI) following coronary artery bypass grafting (CABG).

METHODSThe clinic data of 2242 patients undertaking CABG between July 1997 and July 2006 were retrospectively analyzed, and ARI following CABG was included.

RESULTSARI occurred in 219 patients, with an incidence of 9.8%. Univariate analysis revealed that advanced age, diabetes mellitus, preoperative chronic renal dysfunction, left main disease, low left ventricular erection faction, emergency operation, on-pump CABG, ascending aortic atherosclerosis, postoperative respiratory function insufficiency and low cardiac output syndrome were significantly related to ARI following CABG, and logistic multivariate regression analysis showed that presence of advanced age (P = 0.031), preoperatively chronic renal dysfunction (CrCl or= 150 micromol/L, P = 0.041), on-pump CABG (P < 0.001), postoperative respiratory function insufficiency (P = 0.013) and low cardiac output syndrome (P = 0.004) were independent risk factors of ARI.

CONCLUSIONSAdvanced age, preoperatively chronic renal dysfunction, on-pump CABG, postoperative respiratory function insufficiency and low cardiac output syndrome are the risk factors of ARI following CABG.

Acute Kidney Injury ; epidemiology ; etiology ; China ; epidemiology ; Coronary Artery Bypass ; adverse effects ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; etiology ; Retrospective Studies ; Risk Factors

OBJECTIVETo explore an ideal skin substitute with its appearance and texture similar to normal skin, to repair wounds with full-thickness skin defect.

METHODSComposite skin (CS) in question was composed of allo/xenogeneic acellular dermal matrix (ADM) and razor thin autoskin. One step skin grafting was employed in the experimental study and clinical trial. Razor thin autoskin alone was used as the control in the study. Changes in the antigenicity of ADM and the reformation of basement membrane (BM) structure at epidermis-dermis junction (EDJ) of ADMs were studied at designated time points after the grafting with biochemical and immunohistochemical methods. Fifty-three patients with full thickness skin defects due to various causes, including scar excision were grafted with CS, and survival rate and long-term result were observed.

RESULTSThe grafted CS survived satisfactory. The reformation of the basement membrane structure was clearly observed at the 28th post-graft week. The basement membrane cells grew with polarization in an undulating arrangement. There was reformation of dermal papillae and ridges. The antigenicity of allo-ADM was obviously lower than that of xeno-ADM. Sixty-five out of 70 pieces of CS grafting (92.9%) survived totally, two of them survived partially, and three failed due to infection. The longest follow-up period was 8 and a half years. The grafted CS appeared similar to the normal skin in regard to the texture and color, especially allo-ADM, and no evident rejection reaction was seen.

CONCLUSIONADM possessed very low antigenicity, thus serving a lasting framework after grafting. In addition, it could serve as a "dermal template" for the induction of tissue regeneration.

Animals ; Burns ; surgery ; Dermis ; transplantation ; Follow-Up Studies ; Graft Survival ; Humans ; Male ; Rabbits ; Skin Transplantation ; methods ; Swine ; Transplantation, Autologous ; Transplantation, Heterologous ; Transplantation, Homologous ; Treatment Outcome ; Wound Healing

OBJECTIVETo establish a rat model of scalding with controllable depth and area by high pressure steam.

METHODSHigh pressure steam apparatus consisting of an autoclave and a self-made scalding frame was employed in the study. The rats were inflicted with scalding with 0.12 Mpa (1 Mpa = 7500 mmHg) high pressure steam on the back through a hole of 2.6 cm in diameter for 3, 4, 5, 6, 7, 8, 9 and 10 seconds, with five wounds at each time point. The tissue samples were harvested at 24 post injury hour (PIH) for pathomorphological examination. The depth of scald was measured, and injury to the sweat gland and hair follicles injury, the hair growth in scalded area, and the wound healing condition were observed with Photoshop software.

RESULTSThere was positive correlation between the scalding depth and scalding time. The injury time for superficial and deep partial thickness burn and full thickness burn were 3, 5 and 7 seconds respectively. The wound healing time was similar even the scalding became more and more serious when injury time increased from 7 to 10 seconds.

CONCLUSIONThe scalding depth and area in this model could be controlled, and the degree of scald injury could be graded accurately with easy manipulation. The result showed that it was an ideal model of skin burn wound.

Animals ; Burns ; pathology ; Disease Models, Animal ; Male ; Pressure ; Rats ; Rats, Sprague-Dawley ; Steam