OBJECTIVETo investigate the characteristics of PCNA expression in hypertrophic scars and chronic ulcers and to discuss its relation to their formation.

METHODSThe expressive quantity and sites of PCNA were detected with the immunohistochemical SP method.

RESULTSPCNA 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.

CONCLUSIONSThe 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.

Adult ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Female ; Humans ; Male ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Skin Ulcer ; metabolism ; pathology

Purpose　The aim is to sulfonize Hunai polysaccharide fr om p leurotus tuber-rigium(Fr.)Sing. and to evaluate the antioxidative activities of the sulfated po lysaccharide (S-HNP).Methods　S-HNP was prepared by the reacti on of Hunai polysaccharide with chlorosulfonic acid-Pyridine. The antioxidative activities o f S-HNP were evaluated as follows: (1) the inhibition effects on Fe2+- Vc inducing the injury of rat liver mitochondria in vitro, (2) the protectiv e ef fect on CuSO4 -Phen-Vc-H2O2 inducing the damage of DNA, (3) the scaven ging effect on O*-2. Results　 S-HNP could protect mitochondria from lipid peroxidation induced by Fe2+-Vc, i ncluding the inhibitions of the increase of TBARS content, the swelling of mitoc hondria and the decrease of membrane fluidity, and protect DNA from the damage induced by CuSO4-Phen-Vc-H2O2， and scavenge O*-2 generated in the sel f-oxidation of pyrogallic acid. Conclusion　S-HNP exhibi ted marked antioxidative activities.

OBJECTIVE: To explore the expression of mRNA and its protein in burned rats and their effects on burn wound healing. METHODS: A partial-thickness burn of 30% total body surface ar ea was created on the back of 40 Wistar rats. In situ hybridization and immunohi stochemical methods were used to evaluate the location and the amount of the c-fos mRNA and its protein in normal skin and the burned skin, respectively, at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d after burn. RESULTS: Under a light microscope, both the expression of c-fo s mRNA and its protein could be found in the normal skin, but their induction le vels were much higher in the burned skin. The level of fos protein expression reached peak at 3 h after burn while that of c-fos mRNA reached peak at 6 h aft er burn. CONCLUSIONS: The expression of c-fos can be induced by burns. And the peak level expression of c-fos mRNA comes later than that of c-fos p rotein. It indicates that the action of fos protein is induced by post-translat ional modification of pre-existing fos molecules.

To investigate the effects of miRNA-10b(miR-10b)on breast cancer proliferation and apoptosis in MCF-7 and MDA-MB-231, the miR-10b mimics used to increase the endogenous expression of miR-10b, and miR-10b inhibitor used to decrease the endogenous expression of miR-10b, were stably transfected into MCF-7 and MDA-MB-231 breast cancer cells. The expression level of miR-10b was determined by real-time PCR. The effects of miR-10b on proliferation were evaluated by MTT assay, while cell cycle assay and the apoptosis rate were measured by flow cytometry. Bioinformatic software was used to predict the potential targets of miR-10b, and 3′UTR luciferase reporter and qRT-PCR assay were used to verify a direct target of miR-10b. The expression levels of caspase-3 and p21 protein were measured by Western blot. Results confirmed that over-expression of miR-10b could promote the proliferation of breast cancer cells and inhibit the apoptosis by up-regulating the endogenous miR-10b, while the miR-10b inhibitor could restrain the proliferation of breast cancer cells and increase the apoptosis by reducing the endogenous miR-10b. In conclusion, miR-10b could negatively regulate the expression of caspase-3 and p21 by targeting TP53INP1, hence highlighting its potential as an oncogene in breast cancer cells.

OBJECTIVETo investigate the expression sequence and distribution characteristics of the protooncogenes c-fos, c-myc and endogenous basic fibroblast growth factor (bFGF) genes in burned tissues, and to explore the possible effects of changes in these genes' functions on wound healing.

METHODSPartial-thickness burns of 30% TBSA were established on backs of Wistar rats. In situ hybridization and histological methods were used to detect expression of c-fos, c-myc and bFGF genes in normal and burned tissue at 3 h, 6 h, 1 d, 3 d, 7 d and 14 d postburn.

RESULTSAlthough expression of c-fos and c-myc genes and bFGF gene could be found in normal skin, the expression of all three were markedly induced by burn wounds and the expression models in sequence and distribution were quite different. Expression of c-fos gene increased and peaked at 6 h. Signals were mainly localized in both nuclei of dermal fibroblasts and monocytes. The expression of bFGF gene increased at 6 h and peaked at 1 d postburn, and was distributed in the cytoplasm of fibroblasts. C-myc gene peaked 3 d postburn and was also distributed in the cytoplasm of fibroblasts.

CONCLUSIONSThese results indicated that thermal injury could induce the expression of c-fos, c-myc and bFGF at gene level, showing phasic control and regional distribution. The phasic expression of these genes suggests that there is an interaction between protooncogenes and bFGF, which may play an important role in wound healing. The different expressions of c-fos and c-myc play an inducing role in regulating bFGF, and in turn affect wound healing.

Animals ; Burns ; metabolism ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation ; Genes, fos ; Genes, myc ; In Situ Hybridization ; Male ; Rats ; Rats, Wistar ; Time Factors

OBJECTIVE To investigate the effect of Banxia Xiexin decoction(BXD) on colitis associated cancer(CAC) mice and its related mechanism. METHODS Seventy-five C57BL/6 mice were randomly divided into normal group, model group, Banxia Xiexin decoction low-dose group, high-dose group and mesalazine group. Except for the normal group, the mice in the other groups were intraperitoneally injected with azoxymethane combined with oral dextran sodium sulfate to establish the CAC model. BXD and mesalazine were given respectively for intervention. The general conditions of all mice were observed and recorded, and the changes of body weight, disease activity index, colon length and tumor number were monitored. HE staining was utilized to observe the pathological changes of colon tissue. The expression levels of PCNA, NF-κB P65 and IκB-α were detected by immunohistochemistry. The mRNA levels of IL-17A, N-cadherin, E-cadherin and Bcl-2 were detected by qRT-PCR. Macrophage infiltration was measured using immunostaining analysis. Western blotting was applied to analyze the expression of NF-κB, E-cadherin and N-cadherin proteins in colon tissues of each group. RESULTS There was no significant tumor occurrence in the normal group, while the body weight of the model group mice was significantly reduced and the number of colon tumors increased. The colon length, number of tumors, and degree of inflammatory cell infiltration in the BXD group were significantly improved compared to the model group. Immunohistochemical results showed that the expression of PCNA, NF-κB P65 and IκB-α protein in colon tissue of model group was remarkably increased (P<0.01). Immunofluorescence results showed that the number of F4/80, CD80 and CD206 positive macrophages in the colon tissue of the model group increased (P<0.05 or P<0.01). The results of RT-PCR demonstrated that the levels of IL-17A, N-cadherin and Bcl-2 mRNA in the colon tissue of the model group were significantly increased (P<0.01), while the level of E-cadherin mRNA was fundamentally decreased (P<0.01). Western blotting results displayed that the expression levels of NF-κB and N-cadherin protein in colon tissue of model group were up-regulated (P<0.01), while E-cadherin was significantly down-regulated (P<0.01). Compared with the model group, the changes of the above indexes in the BXD and mesalazine groups were ameliorated, with statistical differences (P<0.05 or P<0.01), and the changes in the BXD high-dose group were more significant. CONCLUSION BXD exhibits strong anti-inflammatory and anti-tumor benefits in CAC mice, inhibiting macrophage activation in colon tissue and promoting M2 polarization, while reducing the expression of tumor associated proteins PCNA and Bcl-2, and block the progression of EMT related proteins (E-cadherin and N-cadherin). The mechanism may connect to suppressing NF-κB P65 and IκB-α activation to regulate the NF-κB signaling pathway.

OBJECTIVE To investigate the effect of Banxia Xiexin decoction(BXD) on colitis associated cancer(CAC) mice and its related mechanism. METHODS Seventy-five C57BL/6 mice were randomly divided into normal group, model group, Banxia Xiexin decoction low-dose group, high-dose group and mesalazine group. Except for the normal group, the mice in the other groups were intraperitoneally injected with azoxymethane combined with oral dextran sodium sulfate to establish the CAC model. BXD and mesalazine were given respectively for intervention. The general conditions of all mice were observed and recorded, and the changes of body weight, disease activity index, colon length and tumor number were monitored. HE staining was utilized to observe the pathological changes of colon tissue. The expression levels of PCNA, NF-κB P65 and IκB-α were detected by immunohistochemistry. The mRNA levels of IL-17A, N-cadherin, E-cadherin and Bcl-2 were detected by qRT-PCR. Macrophage infiltration was measured using immunostaining analysis. Western blotting was applied to analyze the expression of NF-κB, E-cadherin and N-cadherin proteins in colon tissues of each group. RESULTS There was no significant tumor occurrence in the normal group, while the body weight of the model group mice was significantly reduced and the number of colon tumors increased. The colon length, number of tumors, and degree of inflammatory cell infiltration in the BXD group were significantly improved compared to the model group. Immunohistochemical results showed that the expression of PCNA, NF-κB P65 and IκB-α protein in colon tissue of model group was remarkably increased (P<0.01). Immunofluorescence results showed that the number of F4/80, CD80 and CD206 positive macrophages in the colon tissue of the model group increased (P<0.05 or P<0.01). The results of RT-PCR demonstrated that the levels of IL-17A, N-cadherin and Bcl-2 mRNA in the colon tissue of the model group were significantly increased (P<0.01), while the level of E-cadherin mRNA was fundamentally decreased (P<0.01). Western blotting results displayed that the expression levels of NF-κB and N-cadherin protein in colon tissue of model group were up-regulated (P<0.01), while E-cadherin was significantly down-regulated (P<0.01). Compared with the model group, the changes of the above indexes in the BXD and mesalazine groups were ameliorated, with statistical differences (P<0.05 or P<0.01), and the changes in the BXD high-dose group were more significant. CONCLUSION BXD exhibits strong anti-inflammatory and anti-tumor benefits in CAC mice, inhibiting macrophage activation in colon tissue and promoting M2 polarization, while reducing the expression of tumor associated proteins PCNA and Bcl-2, and block the progression of EMT related proteins (E-cadherin and N-cadherin). The mechanism may connect to suppressing NF-κB P65 and IκB-α activation to regulate the NF-κB signaling pathway.