Objective To explore the role of myofibroblasts expressing α-smooth muscle actin (a-SMA) in hyperplastic scar formation.Methods Expression of α-SMA in normal skin and scar was examined by means of immunohistochemistry.The contents of myofibroblasts were measured and com-pared with that of hydroxyproline through cell culture.The contents of precollagen (pc β) , transforming growth factor β (TGF-β) and fibronectin in fibroblasts and myofibroblasts were assayed by ELISA.Results Vascular smooth muscle cells rather than other hypodermal cells expressed α-SMA in normal skin (P<0.01).The expression ofa-SMA was markedly increased up to 96.89% in hyperplastic scar tissues, with statistical difference compared with normal skin (P < 0.01).The contents of hydroxyproline (P <0.01) , pc (P <0.05) , TGF-β (P <0.01) and fibronectin (P <0.05) in myofibroblasts were significantly higher than those in fibroblasts.Conclusion Myofibroblasts play a key role in formation of hypertrophic scar.

With the development of modern medicine,to see a doctor is no longer a simple process of diagnosis or taking drugs.Human have deeply recognized medical science and health from a simple biomedical model to a social,psychological and biomedical model.On the one hand,people seek help from medical science.On the other hand,people fear that malpractice could occur.It has been in a love-hate mentality for people to seek medical and legal relief.In the new era,physician-patient relationship needs to be recognized again from medical staff,patients and legal angles.The combination of medicine and law is the requirement of development of the times.No matter how strong the law is,and no matter how much we hope the legislation is from strength to strength,moderation is still the virtues of the law.Medical perspective is still required in the examination of medical activity.

Objective To investigate the expression changes of paxillin and focal adhesion kinase(FAK) in scar tissues at different periods.Methods The expressions of paxillin and FAK of scar tissue samples from 30 babies at age of 3-12 months were detected by means of immunohistochemistry.Results The expressions of paxillin and FAK in the scar tissues at the 3th and 6th months were increased more significantly than that in normal skin tissues.Whereas,there was no significant difference upon the expressions of paxillin and FAK in the scar tissues at the 12th week compared with normal skin tissues.Conclusion Focal adhesion varies with change of composition,structures and mechanical properties of extracellular matrix(ECM) and with different intracellular framework.Different signal transmissions of ECM,integrin and focal adhesion compound may result in different changes of cellular function.

Objective:To investigate the receptor pathways of glycated basic fibroblast growth factor (bFGF) on proliferation and vascularization of human dermal microvascular endothelial cells (HDMECs).Methods:The experimental research method was used. Glycated bFGF stimulating solution was prepared with glucose and bFGF. Cells of the third to sixth passages were subcultured from the HDMEC primary cell line. The cells were divided into normal control group, glycated bFGF alone group, small interfering RNAs (siRNA)-advanced glycation end product receptor (RAGE) alone group, and siRNA-RAGE+glycated bFGF group, and seeded into 96-well plate with 6 wells in each group, and seeded into 6-well plate with 3 wells in each group. Cells in siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group were transfected with siRNA-RAGE and then were added into HDMEC culture medium for routine culture. After two days, the original HDMEC culture medium was discarded, and cells in siRNA-RAGE alone group were routinely cultured in HDMEC culture medium, cells in siRNA-RAGE+glycated bFGF group were routinely cultured in glycated bFGF stimulating solution. Cells in normal control group were routinely cultured in HDMEC culture medium, and cells in glycated bFGF alone group were routinely cultured in glycated bFGF stimulating solution. After transfection with siRNA-RAGE, cells were seeded into 48-well plate and divided into siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group, with 4 wells in each group. Another cells were directly seeded into 48-well plate without transfection and divided into normal control group and glycated bFGF alone group, with 4 wells in each group. Cells in normal control group and siRNA-RAGE group were routinely cultured in HDMEC culture medium. Cells in siRNA-RAGE alone group and siRNA-RAGE+glycated bFGF group were routinely cultured in glycated bFGF stimulating solution. Cells were divided into normal control group, glycated bFGF alone group, siRNA-fibroblast growth factor receptor (FGFR) alone group, and siRNA-FGFR+glycated bFGF group and seeded into 96-, 6- and 48-well plates. respectively. Cell density, sample number, and corresponding treatment were the same as before. Only siRNA-RAGE was replaced by siRNA-FGFR. Cells were divided into siRNA-positive control group, siRNA-negative control group, siRNA-RAGE group, and siRNA-FGFR group and transfected with siRNA-glyceraldehyde-3-phosphate dehydrogenase positive control, siRNA-negative control, siRNA-RAGE, and siRNA-FGFR for 4 to 6 hours, and then were added into HDMEC culture medium for routine culture. The transfection effect of siRNA was identified by reverse transcription PCR. Cell counting kit 8 method was used to determine the proliferation of cells after 2 days of culture (sample number was 6), flow cytometry was used to detect the apoptosis of cells after 2 days of culture (sample number was 3), tube forming test was used to detect the angiogenesis of cells cultured after 6 hours of culture (sample number was 4). Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results:At the 200 bp band, there were no complementary DNA (cDNA) bands in siRNA-positive control group, siRNA-RAGE group, and siRNA-FGFR group, but cDNA bands were detected in siRNA-negative control group, indicating the success of siRNA transfection. After 2 days of culture, the absorbance value of cells in glycated bFGF alone group was significantly lower than that of normal control group ( t=2.359, P<0.05); absorbance value of cells in siRNA-RAGE+glycated bFGF group was significantly higher than that of glycated bFGF alone group ( t=3.858, P<0.01), which was similar to that of siRNA-RAGE alone group ( t=2.148, P>0.05). The absorbance value of cells in siRNA-FGFR+glycated bFGF group was similar to that of glycated bFGF alone group ( t=0.805, P>0.05), but significantly lower than that of siRNA-FGFR alone group ( t=4.201, P<0.01). After 2 days of culture, the apoptotic rate of cells in glycated bFGF alone group was significantly higher than that of normal control group ( t=2.416, P<0.05). The apoptotic rate of cells in siRNA-RAGE+glycated bFGF group was significantly lower than the rates in glycated bFGF alone group and siRNA-RAGE alone group ( t=3.861, 2.724, P<0.05 or P<0.01). There were no significant differences in apoptosis rate of cells among normal control group, glycated bFGF alone group, siRNA-FGFR alone group, and siRNA-FGFR+glycated bFGF group ( F=2.218, P>0.05). After 6 hours of culture, the number of tubules of cells in normal control group (636±5) was significantly more than that of glycated bFGF alone group [580±8, t=10.825, P<0.01], and the number of tubules of cells in siRNA-RAGE+glycated bFGF group (647±10) was significantly more than those of glycated bFGF alone group and siRNA-RAGE alone group (628±4, t=13.040, 3.641, P<0.01). After 6 hours of culture, the number of tubules of cells in siRNA-FGFR+glycated bFGF group (619±5) was significantly more than that of glycated bFGF alone group ( t=9.000, P<0.01), but less than that of siRNA-FGFR alone group (632±3, t=2.814, P<0.05). Conclusions:Glycated bFGF affects the angiogenesis of HDMEC through RAGE pathway, which may be one of the reasons for the impaired wound healing of diabetic skin.

Objective To identify the factors related to prognosis of diabetic foot ulcer (DFU).Methods A total of 186 patients with type Ⅱ DFU from a single diabetic foot center was included in this prospective study.Follow-up of the final outcome (healing,major amputation or death) was made in 6 months.Influence of patient demographics and clinical data on outcome was detected using multivariate Logistic regression analysis.Results Follow-up was performed in 172 patients,of whom 147 were cured (55 cases cured after minor amputations),3 underwent major amputations,6 died,and 16 were not yet cured at the final follow-up.In multivariate Logistic regression analysis,the outcome was independently correlated with ischemia (P ＜0.01),infection (P ＜ 0.05),ulcer number (P ＜ 0.01) and peripheral neuropathy (P ＜ 0.05) ; the risk of poor outcome increased with ischemia [odds ratio (OR) =10.8],infection (OR =211.4),ulcer number (OR =39.5),and peripheral neuropathy (OR =181.1).Conclusion Prognosis of DFU is associated with ischemia,infection,ulcer number,and peripheral neuropathy.

Objective To assess the deep inferior epigastric perforator (DIEP) vessels in patients with breast reconstruction flaps by contrast-enhanced ultrasound (CEUS) and three-dimensional ultrasound (3DUS) reconstruction techniques.Methods Conventional ultrasound,CEUS and 3DUS were used to evaluate DIEP vessels of breast reconstruction flaps in 43 patients before surgical procedures.The identification,localization and spatial relationship of DIEP vessels were analyzed with conventional ultrasound,CEUS and 3DUS methods.The findings of CEUS and 3DUS were compared to conventional ultrasound and surgical outcome.Results Compared to CDFI,40 cases (93％) were observed more clearly with CEUS,and were showed more accurately than conventional ultrasound.41 cases (95％) could be displayed wonderfully in 3D ultrasound.Perforators which were detected by ultrasound were confirmed in the surgery and the transferred flaps survived completely.Conclusions Perforators can be displayed more clearly and located more accurately by CEUS and 3DUS.CEUS and 3DUS could play a very important role in the preoperative navigation of the DIEP than conventional ultrasound.

Objective To study the effect of mechanical tension on collagen arrangement and illustrate the relationship between tissue architecture and tension properties. Methods Cell morphologies, orientation and collagen arrangement of fibroblasts cultured in three different types of collagen gels with variation of mechanical tension were observed by phase contrast photomicrographs, light microscopy and transmission electron microscopy. Expression and distribution of a-smooth muscle actin (α-SMA) were investigated by immunohistochemistry. Results Phase contrast photomicrographs, light microscopy and transmission electron microscopy showed high level of tension distributed anisotropically in the monolayer gels and the anchored collagen gels, with bipolar shape of the fibroblasts, obvious polarity, arrangement of exogenous collagen fibres parallel to the long axis of the fibroblasts, especially prominent in monolayer gels. Low level of tension distributed isotropically was observed in floating collagen gels, with stellate morphology and arrangement of exogenous collagen fibres in a reticular array. Immunofluorescence showed that fibroblasts expressed high level of α-SMA protein distributed along the long axis of fibroblasts in the monolayer gels and the anchored collagen gels, especially in former ones. In contrast, few expression of α-SMA protein was found in floating collagen gels. Cell morphologies and orientation, expression and distribution of α-SMA as well as collagen arrangement of fibroblasts in the monolayer gels and the anchored collagen gels were similar to those in granulation tissue, whereas floating collagen gels resembled normal dermis or remodelled tissues. Conclusions Tissue architecture or morphology of the dermis are corresponding to tension proporties. Different tissue architectures are closely correlated with particular tension proporties.

Objective To detect the histological characteristics of collagen nodules from hypertrophic scars (HS) and investigate the origin of collagen nodules.Methods The scar tissues were collected from patients with plastic operation.Morphological characteristics of collagen nodules were observed by light microscopy of HE-stained sections; expressions of type Ⅰ /Ⅲ collagens were observed by polarized light microscopy of sirius red-stained sections; expression and distribution of myofibroblasts (MFb)-specific protein (α-smooth muscle actin,α-SMA) were observed by immunostaining in order to observe level of local tissue tension.Results Collagen nodules varied in shape,not only sphereshaped,and in size.Moreover,abundant fibroblasts (Fb) with large and light-stained nuclei were seen in the nodules compared to non-nodule area,indicating that the cells located in the modules were active.Some collagen nodules were composed largely of collagen type Ⅲ (green),but some mainly contained collagen type Ⅰ (red or yellow),indicating the difference in the time of nodule formation.α-SMA was expressed mainly in the deep dermis equivalent to the level of reticular layer of the new scar tissues (2 months after injury) ; α-SMA was expressed mainly in the nodules of the old scar tissues (2-10 years after injury),but almost not in non-nodule areas except for a strongly positive staining in the vessels.Moreover,α-SMA presented a heterogeneous distribution in the collagen nodules,with stronger expression in the epidermal end than in the subcutaneous tissue end and stronger expression in the superficial dermis than that in the deeper part.It was suggested that there existed massive amount of MFb and high tension in the nodules arid that the tension distribution was not uniform in or between the nodules.Conclusions Collagen nodules are of varying shape and size and collagen types are associated with the time of nodule formation.Moreover,Formation of the collagen nodules may be closely related to the distribution and evolution of the local tissue tension.

AIM: To explore the histochemical changes o f diabetic skin and the pathogenesis of impaired wound healing in diabetes. METHODS: 54 male Sprague-Dawley (SD) rats weighing 200-220 g wer e randomized into control and STZ-induced diabetic groups. The shaved skin speci mens from the back of rats were collected in 4, 8 and 12 weeks post STZ-inductio n, respectively. Hematoxylin-eosin dye was used for histological examination. Me a nwhile, the skin glucose contents were measured by Beckman's autoanalyzer. Skin AGEs concentrations were assessed by detecting total fluorescence in tissue coll agen and immunohistochemistry assay. RESULTS: The skin thickness in diabetic animals was decreased, w ith the features of multilayer epithelium structure disappeared in epidermis and collagen fibers atrophied, swollen and degenerated in dermis; The inflammatory responses in the dermis of diabetic animals were increased obviously. The result s also revealed that skin glucose contents in diabetic rats [(2.64?1.03)mg/ g skin] were 2-3 times higher than those in the controls [(0.74?0.33)mg/g s kin] (P

Objective To identify the cross-reactive antigens shared by Mycobacteria smegmatis(MS) and Mycobacteria tuberculosis(MTB) and to analyze their antigenicity.Methods Bacterial antigens were extracted from strains of MS and MTB by ultrasonication.Western blot assay was performed to analyze common antigens that reacted with both of the antiserum samples against MS and MTB.The extracted bacterial antigens were mixed with incomplete Freund′s adjuvant and then were injected into muscles of mice.Cytokines secreted by murine spleen lymphocytes following stimulation with various antigens of MS and MTB were determined by ELISPOT and flow cytometry on the 7th day.IgG levels in serum samples were detected by ELISA 7 days after injection.Results There were cross-reactive antigens shared by MS and MTB.Potent humoral immune responses and cellular immunity against both MS and MTB could be induced by those cross-reactive antigens after sensitization the mice by either MS or MTB antigens.Cytokines of IL-2 and IFN-γ in CD4+ and CD8+T cells of mice stimulated with MS or MTB antigens were significantly increased as compared with those of non-sensitization group and those of Brucella antigens stimulation group.ConclusionCross-reactive antigens shared by MS and MTS can effectively promote specific immune reactions to the infection of MTB, which provides a scientific basis for the development of tuberculosis vaccines.