Bronchial asthma is the most common chronic disease in children.In recent years,the inci-dence of bronchial asthma is rising and seriously harmed to the public health,has been gained comprehensive attention in community and scholars.With the in-depth study of the pathogenesis of asthma,the immunologi-cal dysfunction caused by disordered Th1 /Th2 cell play a major role in the pathogenesis of asthma,and im-munotherapy for its pathogenesis has also made great progress,which is expected to become the safer,more effective and more specific treatment.Currently immunomodulator for the treatment of asthma can be divided into four categories according to the source:microbial agents,biological agents,herbal or plant origin and syn-thetic agents.Microbial agents which include BCG,Pan Fu Shu,biologics including Th2 receptor antagonist, Th1 cytokines,anti-IgE antibody,immunoglobulin,thymosin,synthetic agents include pidotimod,transfer fac-tor,traditional Chinese medicine or plant.The main source of Gin kgo biloba extract,Huaiqihuang etc.We make a summary on the research progress and immunomodulator in the treatment of asthma in this review.

nal abnormalities of pediatric heart diseases.

Objective To investigate the effect of bFGF and MSCs autografting on expression of PCNA and Ⅷ-R Ag of porcine skin defect wound. Methods Six male minipigs were used to produce 48 full-thickness skin wounds, each wound area about 2.54 cm~ 2 on both sides of the back (8 wounds per pig). These 48 wounds were randomly treated with bFGF, MSCs, bFGF+MSCs, and normal saline (12 wounds for each treatment). The tissue from the wounds was harvested on day 3, 7, 14 after surgery, then used for HE staining and immunohistochemical staining for proliferating cell nuclear antigen (PCNA) and factor Ⅷ related antigen (Ⅷ-R Ag). Results Granulation tissue was presented in all groups on day 3 and 7 after wound, most abundant in the wounds treated with bFGF+MSCs, as well as blood capillary formation. PCNA expression increased after skin injury, reached the peak on day 7, most in the granulation tissue, and lasted to day 14, then began to decrease. On day 3, 7, 14 after skin injury, PCNA expression was the most highest in the wounds treated with bFGF+MSCs. Ⅷ-R Ag-positive cells in the wounds treated with bFGF+MSCs were significantly more than that of the other three treatments (P

Objective To investigate the influencing conditions of human epidermal stem cells(ESCs) differentiating into gland-like cells and to identify the induced tubular structure.Methods The ESCs were seeded onto compound polysaccharide shell dermal matrix and collagen gel,adding different concentrations of epidermal growth factor(EGF) and culturing by vertical shaking in vitro,the three dimensional culture and induced directional differentiation were performed.The means of HE staining,immunofluorescence and confocal laser scanning microscope were adopted to observe the conditions,morphology and phenotype change of ESCs directionally differentiating into glandular epithelium-like cells.Results 15-20 ng/mL EGF could induce ESCs in tissue-engineering dermis to grow into dermis and appear the gland-like structure.The HE staining in this structure showed its profile as a single layer with lacune in the middle compartment,eosinophilic cytoplasm,lightly stained.Under CLSM,by CK19 staining,the luminal structure in the middle of the cell mass was observed,while CK18 and CEA were expressed in this structure.Conclusion Under the induction of particular concentration of EGF,the in vitro cultured human ESCs seeded on the tissues engineering dermis could form into the tubular structure,which is similar to the in vivo sweat gland secretory cells in morphology and histology.

Objective To investigate whether epithelial-mesenchymal transition (EMT)is involved in the anti-invasion and anti-metastasis effects of curcumin on MDA-MB-2 3 1 breast cancer cell line and further analyze the underlying mechanisms.Methods MTT method was used to detect the anti-proliferative effect of curcumin on MDA-MB-2 3 1 cell line induced by lipopolysaccharide (LPS).The morphological changes were determined by optical and transmission electron microscopy,respectively.The expressions of Vimentin,E-cadherin,NF-κB and Snail were analyzed using RT-PCR and Western blotting.Results Curcumin could inhibit the occurrence of LPS-induced EMT of MDA-MB-2 3 1 breast cancer cell line, decrease the expression of LPS-induced EMT marker Vimentin and increase the expression of E-cadherin, resulting in the inhibition of in vitro cell motility and invasiveness.These actions were mediated by inactivating NF-κB-Snail signaling pathways.Conclusion Our data provide a new perspective of the anti-invasion mechanism of curcumin,indicating that the effect is in part due to its ability to inhibit the EMT process.

Objective To investigate the effects of peptide-binding domain (PBD) of heat shock protein (HSP) 72 on epithelial to mesenchymal transition (EMT) in rat renal tubular epithelial cells.Methods The expressions of wild-type HSP72,mutant of HSP72 lacking peptide binding domain (HSP72-△PBD) and HSP72-PBD were induced by plasmid transfection.NRK-52E ceils were stimulated by TGF-β1 for 48 h.The expressions of α-smooth muscle actin (α-SMA),E-cadherin,HSP72 and Smad3/p-Smad3 were detected by Western blot and immunofluorescence.Results After NRK-52E cells were stimulated by TGF-β 1 (10 μg/L) for 48 h,the expression of α-SMA was increased and the protein level of E-cadherin was decreased.Western blotting and immunofluorescence showed that over-expression of both HSP72 and PBD inhibited TGF-β1-induced up-regulation of protein α-SMA expression,down-regulation of protein E-cadherin.However,overexpression of HSP72-△PBD did not change the protein level of E-cadherin and α-SMA.In addition,over-expression of HSP72 and PBD significantly inhibited the phosphorylation of Smad3.Conclusion Inhibition of Smad3 activation and EMT by HSP72 is associated with the function of PBD.

Objective To compare the epidermal shape built by different passages of keratinocytes and its ability of proliferation and differentiation in three-dimensional conditions.Methods Different passages of keratinocytes were used to construct tissue-engineered skin.The morphology of the tissue-engineered skin was observed with HE and PAS staining,while CK1/CK10,CK5/CK14,Ki67 were detected by immunohistochemical assays.Results All the tissue-engineered skin had a significant dermoepidermal structure.The stratification of 1st and 2nd passage skins were better,and 2nd passage epidermis was thicker than that in other passages (P＜0.05).Dermoepidermal structure in collagen type Ⅳ group binded more tightly,but collagen type Ⅳ had little effect on the thickness of the epidermis (P＞0.05).In collagen type Ⅳ group PAS stain was negative,indicating type Ⅳ collagen was unable to promote the reconstruction of BM in vitro.The Ki-67 proliferation index of the 2nd keratinocyte was similar to the normal skin,the remaining passages keratinocyte proliferation gradually decreased (P＜0.05) ; the 1st and 2nd passage skins expressed CK1/CK10 and CK5/CK14.Conclusions Keratinocytes before the 3rd passage have a better ability in the proliferation and differentiation,and so they are more suitable as seed cells for tissue-engineered skin.

Objective To investigate the role of protein kinase C (PKC) in induction of vascular endothelial growth factor (VEGF) secretion by isoflurane in primary cultured rat cardiomyocytes. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into 6 groups ( n = 6 each): control group (group C), 3 different concentration isoflurane groups (group Ⅰ1-3 ), PKC inhibitor calphostin C group (group P), and PKC inhibitor + isoflurane group (group PI). The cells were exposed to 0.7%, 1.4% and 2.1% isoflurane for6 h in group Ⅰ1-3 respectivly. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L in group P. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L, then the cells were exposed to 1.4% isoflurane for 6 h in group PI. VEGF concentrations and expression of PKC isoforms were determined by ELISA and Western blot respectively. Results Compared with group C, the VEGF concentration was significantly increased in group Ⅰ2 and Ⅰ3, and PKCε expression was down-regulated in the cytoplasm while upregulated in the cytomembrane in group Ⅰ2 ( P ＜ 0.01 ), but no significant change was found in the parameters mentioned above in group Ⅰ2 ( P ＞ 0.05). PKCα, PKCδ and PKCζ expression was significantly higher in the cytoplasm than in the cytomembrane in group C and Ⅰ2. VEGF concentrations were gradually increased with the increase in isoflurane concentrations ( P ＜ 0.05). VEGF concentrations were significantly lower in group PI than in Ⅰ2 ( P ＜0.05) .Conclusion Isoflurane induces VEGF secretion in primary cultured rat cardiomyocytes through translocation of PKCε from the cytoplasm to the cytomembrane, suggesting that it is a mechanism of the cardioprotective effects of isoflurane.

Objective To investigate the clinical outcomes of surgical excision combined with recom binant interferon aipha-2b in the treatment of acral malignant melanoma (MM). Methods Fifteen patients with acral MM admitted to the department since 2004 were recruited into this study. The tumors varied from 1.8 mm to 3.9 mm in invasion depth. Thin tumors with an invasion depth of 1.8 - 2.0 mm were excised with a margin of lcm beyond the tumors, and those with an invasion depth of 2.0 - 3.9 mm were excised with a margin of 2 cm beyond the tumors. After excision, 4 cases of minor excision were sutured directly, 10 cases of large excision were repaired with adjacent skin grafts and flaps, 1 patient with the involvement of the first metatarsophalangeal joint underwent toe amputation followed by the repair of planta wound surface with the remaining skin on the dorsa of toes. Patients received intramuscular recombinant interferon alpha-2b for 3 months (3 million units daily for the first 3 days and 6 million units for the remaining days) following operation. Results There were 6 cases of MM in situ and 9 cases of invasive MM in this study. All the skin grafts and flaps survived. Within the 3-year follow up, relapse was observed only in 1 patient with invasive MM. Recovery was achieved in the functions of feet in all patients. Conclusion The excision of tumors with a margin determined by tumor thickness plus intramuscular interferon alpha-2h may improve the survival of patients with cutaneous MM in planta pedis with avoidance of amputation.

Objective To study histologic characteristics of bilayer skin substitute reconstructed by cells from human hair follicle and whether the skin appendage can be induced based on the bilayer skin substitute.Methods After composite chitosan bilayer skin substitute was reconstructed with dermal papilla cells and outer root sheath cells or dermal sheath cells and outer root sheath cells,its histologic characteristics was investigated in vitro and after transplanted onto SD albino rats.Results Composite chitosan bilayer skin substitute reconstructed by cells from hair follicle had closely arranged epithelium cells and outstanding cornification;Epithelial cords linked with epidermis could be seen in dermis.However,there was no certain hair follicle-like structure formation either in vitro or in vivo.Conclusion Hair follicle cells are good source for skin substitute reconstruction,but it can not induce skin appendage formation through skin substitute by now.