Effect of pomegranate peel polyphenol gel on cutaneous wound healing in alloxan-induced diabetic rats.

BACKGROUND: Pomegranate (punica granatum) belongs to the family Punicaceae, and its peel has been used as a traditional Chinese medicine because of its efficacy in restraining intestine, promoting hemostasis, and killing parasites. Pomegranate peel has been reported to possess wound-healing properties which are mainly attributed to its polyphenol extracts. The purpose of this study was to investigate the effect of pomegranate peel polyphenols (PPP) gel on cutaneous wound healing in diabetic rats. METHODS: Alloxan-induced diabetic rats were given incisional wounds on each side of the mid-back and then treated daily with PPP gel (polyphenol mass fraction = 30%) post-wounding. Rats were sacrificed on days 4, 7, 14, and 21 post-wounding to assess the rates of wound closure, histological characteristics; and to detect the contents of hydroxyproline, production of nitric oxide (NO), and activities of NO synthase (NOS), as well as the expressions of transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) in wound tissue. RESULTS: Wound closure was significantly shortened when PPP gel was applied to the wounds of diabetic rats. Histological examination showed the ability of PPP gel to increase fibroblast infiltration, collagen regeneration, vascularization, and epithelialization in the wound area of diabetic rats. In addition, PPP gel-treated diabetic rats showed increased contents of hydroxyproline, production of NO, and activities of NOS and increased expressions of TGF-ß1, VEGF, and EGF in wound tissues. CONCLUSION: PPP gel may be a beneficial method for treating wound disorders associated with diabetes.

[Study on the antimutagenicity of Cordyceps taii polysaccharide].

OBJECTIVE: To study the antimutagenicity of Cordyceps taii polysaccharide (CDPl). METHODS: Ames test and micronucleus form assay of polychromatic erythrocytes in mice bone marrow were used to evaluate the antimutagenic effect of CDPI with different concentrations. RESULTS: CDP1 could inhibit the reverse mutation on Salmonella typhimuriun strains TA97, TA98, TA100 and TA102 without S9 in different degree, especially remarkable in the strains T97 ,T98 and T102. The maxium inhibition rates of strains TA97, TA98, TA100 and T102 were 68.5%, 79.5%,19. 8% and 81.3%, respectively. In addition, CDPI could significantly reduce micronucleus frequencies induced by cyclophosphamide, and the rate of maxium inhibition was 70.3%. CONCLUSION: Cordyceps taii polysaccharide possesses significantly antimutagenic properties.

[Effect of decreased GSH on sensitivity of breast cancer cells to ADM].

OBJECTIVE: Glutathione(GSH) maintains an optimum cellular redox potential. Elevated levels of GSH render some types of cancer cells resistant against anti-cancer drugs. The aim of this study was to determine the effect of a thiol-depleting agent, diethylmaleate (DEM), on the sensitivity of human breast cancer cells to ADM. METHODS: The ADM-resistant human breast cancer MCF-7/ADM cell lines and ADM-sensitive MCF-7/S cell lines were treated by thiol-depleting agent DEM for 3 h respectively. The changes of sensitivity to ADM were then measured by MTT assay. The intracellular GSH contents were examined by fluorescent-spectrophotometry and the correlation between the changes of sensitivity to ADM and the intracellular GSH content was analyzed. RESULTS: Treatment of MCF-7/ADM and MCF-7/S cells by 0.1 micromol/L DEM for 3 h decreased 37.4% and 29.7% of the intracellular GSH content respectively (P < 0.01). ADM also decreased intracellular GSH content in a ADM-concentration-dependent manner. The combined use of DEM and ADM depleted the intracellular GSH content in both cells significantly more than the sum of single use of ADM and DEM alone. The sensitivity of both cells to ADM increased with the decline of intracellular GSH content. CONCLUSION: The depletion effect of DEM on the intracellular GSH could be enhanced by ADM and such depletion may be involved in the changes of the sensitivity of MCF/7 cells to ADM.

[The abnormality of FcgammaRIIB1-mediated signaling and the hyperactivity of B cells from patients with systemic lupus erythematosus].

AIM: To evaluate the effect of FcgammaRIIB(1) (CD32) representative self-inhibitory adjustive mechanism of B cells on pathogenesis of systemic lupus erythematosus (SLE) by observing the expression characteristic and functional state of molecules on the surface of B cells from SLE patients. METHODS: The peripheral blood mononuclear cells (PBMC) were prepared by density gradient centrifugation, and the B cells were isolated from PBMC by magnetic activated cell sorting (MACS). The fluxes of intracytoplasmic calcium ([Ca(2+)](i)) of B cells activated by different activators were measured by fluorescence spectrophotometric method. The IgG production by B cells cultured with activators was assayed by ELISA. The expression levels of CD32, CD19, and IgM on the surface of B cells were measured by flow cytometry. RESULTS: (1)After B cells were stimulated with goat anti-human mu chain F(ab')(2) fragments and whole IgG respectively, the ratio of [Ca(2+)](i) response by F(ab')(2) fragments to whole IgG was significantly lower in SLE B cells compared to rheumatoid arthritis (RA)(P<0.05) or normal (P<0.01) B cells. (2)The ratio of total IgG production by B cells cultured with staphylococcal protein A (SPA) to SPA plus IgG anti-mu chain was significantly lower in SLE patients compared to RA patients or normal individuals (P<0.05). (3)There was no obvious difference in the expression of CD19, CD32, and IgM on the surface of B cells from SLE, RA patients and normal individuals (P>0.05). CONCLUSION: The inhibitory signaling abnormality of CD32 possibly contributes to the mechanism of hyperactivity of human SLE B cells.

Effect of ozone produced from antibody-catalyzed water oxidation on pathogenesis of atherosclerosis.

Recent studies have suggested that antibodies can catalyze the generation of unknown oxidants including hydrogen peroxide (H2O2) and ozone (O3) from singlet oxygen (1O2) and water. This study is aimed to detect the effect of antibody-catalyzed water oxidation on atherosclerosis. Our results showed that both H2O2 and O3 were produced in human leukemia THP-1 monocytes incubated with human immunoglobulin G and phorbol myristate acetate. In the THP-1 monocytes incubated with human immunoglobulin G, phorbol myristate acetate and low density lipoprotein, the intracellular total cholesterol, free cholesterol, cholesteryl ester and lipid peroxides clearly increased, and a larger number of foam cells were observed by oil red O staining. The accumulation of all intracellular lipids was significantly inhibited by vinylbenzoic acid, and only slightly affected by catalase. These findings suggested that the production of O3, rather than H2O2, might be involved in the pathogenesis of atherosclerosis through the antibody-catalyzed water oxidation pathway.

[Antibody-induced ozone production and its potential effects on pathogenesis of atherosclerosis].

In the present study, we measured the antibody-catalyzed 03 formation from THP-1 monocytes activated by phorbol myristate acetate (PMA) by indigo carmine bleaching reaction test, and the accumulation of cholesterol in THP-1 monocytes by fluorescence spectrophotometric method, and analyzed the cholesterol ozonation product 5,6-secosterol by high-performance liquid chromatography (HPLC), to explore the potential effect of antibody-catalyzed water oxidation on pathogenesis of atherosclerosis. It was showed that THP-1 monocytes incubated with human IgG and PMA evidently produced an oxidant with the chemical signature of 03 which could bleach indigo carmine, and be intensified or inhibited respectively by catalase and vinylbenzoic acid. In the THP-1 monocytes incubated with human IgG, PMA and LDL, the intracellular accumulated total cholesterol (TC), free cholesterol (FC), cholesteryl ester (CE) and the CE/TC increased evidently, and the cholesterol ozonation product 5,6-secosterol was also produced markly, all of that were inhibited by vinylbenzoic acid. These results demonstrated that the activated THP-1 monocytes possess the ability to produce O3 through antibody-catalyzed water-oxidation pathway, which could be a new mechanism concerned with atheriosclerosis.

[A novel fluorophotometric method for assaying free glutathione of both reduced and oxidized in plasma].

To investigate the measurement of free glutathione of both reduced or oxidized (i.e. GSH and GSSG) in plasma, and to evaluate the redox state of GGSH/GSSG in human plasma, both GSH and GSSG in plasma were measured by fluorophotometry based on the facts that one molar GSSG can be reduced to two molar GSH by dithiothreitol(DTT) under the condition of the pH being about 6.0, and GSH can provide both primary amine and thiol groups to react with the two carbonyl groups of O-phthaldehyde (OPA) to form a fluorescent ternary isoindole complex at pH 8.0. This method can at least measure 16 picomole GSH and 8 picomole GSSG respectively in the tube. The variation coefficient (CV) for intra-ssay and intera-ssay is about 4. 6% and 3.9% for GSH and 3.5% and 4.1% for GSSG respectively. The recovery of GSH and GSSG added to the plasma is (99.77% +/- 5.70)% and (99.28% +/- 4.73)% respectively. The concentration of GSH and GSSG in the plasma of young healthy volunteer is (16.5 +/- 2.4) nmol x mL(-1) and (1.7 +/- 0.35)nmol x mL(-1) respectively, without significant difference between male and female. This measurement method is simple with great sensitivity and selectivity for rapid measuring GSH and GSSG in human plasma simultaneously.