Scale-Up Synthesis and In Vivo Anti-Tumor Activity of Curcumin Diethyl Disuccinate, an Ester Prodrug of Curcumin, in HepG2-Xenograft Mice.

Previously, we synthesized curcumin and a succinate ester prodrug of curcumin namely curcumin diethyl disuccinate (CurDD) in the lab scale, which yielded hundred milligrams to few grams of the compounds. CurDD was found to be more stable in a phosphate buffer pH 7.4 and exhibited better cytotoxicity against Caco-2 cells than curcumin. Here, the one-pot syntheses of curcumin and CurDD were scaled up to afford multigram quantities of both compounds for preclinical studies using a 10-L chemical reactor. The key steps for the synthesis of curcumin were the formation of boron-acetylacetone complex and the decomplexation of boron-curcumin complex. The synthesis of CurDD could be achieved via a one-step esterification between curcumin and succinic acid monoethyl ester chloride using 4-(N,N-dimethylamino)pyridine as a catalyst. The synthesized curcumin and CurDD were then investigated and compared for an anti-tumor activity in HepG2-xenograft mice. CurDD could reduce the tumor growth in HepG2-xenograft mice better than curcumin. CurDD also exerted the stronger inhibition on VEGF secretion, COX-2 and Bcl-2 expression and induced higher Bax expression in comparison with curcumin. The results suggest that CurDD is a promising prodrug of curcumin and has a potential to be further developed as a therapeutic agent or an adjuvant for the treatment of hepatocellular carcinoma.

High dose oral vitamin C and mesenchymal stem cells aid wound healing in a diabetic mouse model.

OBJECTIVE: This study sought to determine the effects of oral vitamin C (VitC) and mesenchymal stem cells (MSCs) on wound healing in diabetic nude mice. METHOD: Bilateral, full-skin thickness wounds were created as an in vivo wound model in BALB/C diabetic nude mice. The mice were separated into five groups: control (CON); diabetes mellitus (DM, from a streptozotocin injection); DM treated with MSCs (DM+MSCs); DM treated with VitC (DM+VitC), and DM treated with MSCs and VitC (DM+MSCs+VitC). After wounding, daily oral-feeding of high dose VitC (1.5g/l) was administered, and a single dose of MSCs (1x106 cells) was given topically using matrix gel application to the wounded area. RESULTS: At day seven, the lowest rate of wound healing, in terms of percentage of wound closure, appeared in the DM group, as compared with the CON and all other treatment groups (mean percentage of wound closure and standard deviation), CON=75.94±7.09%; DM=55.65±9.59%; DM+MSCs=78.57±6.46%; DM+VitC=77.52±3.31%; and DM+MSCs+VitC=84.61±2.87%, p≤0.05. At day 14 post-wounding, the combination of oral high dose VitC and MSCs accelerated wound healing (91.44±3.19%, p≤0.05). In addition, the highest capillary density in DM+MSCs+VitC was obtained at 14 days post-wounding (29.49±7.30%, p≤0.05). CONCLUSION: The findings of this study highlight the possibility of using oral high dose VitC in adjunct to MSCs to increase angiogenesis and accelerate diabetic wound healing in an animal model. This novel therapeutic approach should be studied further to test if it could be a useful adjunct of existing therapies to prevent infection and amputation in patients with diabetes.

Combinational Treatment Effect of Tetrahydrocurcumin and Celecoxib on Cervical Cancer Cell-Induced Tumor Growth and Tumor Angiogenesis in Nude Mice.

Background: Tetrahydrocurcumin (THC) demonstrated an anti-cancer and anti-angiogenic effects in cervical cancer. Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, have also shown anticancer effect. However, the combinational treatment effect of THC and celecoxib on tumor growth and tumor angiogenesis, especially, using cervical cancer (CaSki)-implanted nude mice has yet not been reported. Objective: To evaluate the combinational treatment effect of THC and celecoxib on tumor progression and tumor angiogenesis in cervical cancer (CaSki)-implanted nude mice. Material and Method: CaSki cells were inoculated in mice to establish subcutaneous tumors. One month after inoculation, vehicle, THC100 mg/kg, Celecoxib100 mg/kg, or THC50 + Celecoxib50 mg/kg was orally administered every day for 28 consecutive days. The tumor volume was measured every 3-4 days. The microvascular density (MVD) was evaluated using the CD31 expression. VEGF, COX-2, and EGFR expression were also detected by immunohistochemistry. Results: THC, celecoxib, and the combination treatments statistically retarded the tumor volume by 70.40, 65.11 and 77.04%, respectively. The MVD was significantly increased in CaSki + vehicle group, but THC, celecoxib, and the combination treatments markedly attenuated the MVD. VEGF, COX-2, and EGFR were up-regulated in CaSki + vehicle group; however, they were attenuated by THC, celecoxib, and the combination treatments. Conclusion: The combinational treatment effect of THC and celecoxib causing inhibition of tumor growth and tumor angiogenesis via down-regulation of VEGF, COX-2 and EGFR expression. However, this combined treatment did not show the synergistic effect on inhibiting the tumor growth and tumor angiogenesis in cervical cancer (CaSki)-implanted nude mice model.

Molecular mechanisms of curcumin on diabetes-induced endothelial dysfunctions: Txnip, ICAM-1, and NOX2 expressions.

We aim to investigate the effects of curcumin on preventing diabetes-induced vascular inflammation in association with its actions on Txnip, ICAM-1, and NOX2 enzyme expressions. Male Wistar rats were divided into four groups: control (CON), diabetic (DM; streptozotocin (STZ), i.v. 55 mg/kg BW), control-treated with curcumin (CONCUR; 300 mg/kg BW), and diabetes treated with curcumin (DMCUR; 300 mg/kg BW). 12th week after STZ injection, iris blood perfusion, leukocyte adhesion, Txnip, p47phox, and malondialdehyde (MDA) levels were determined by using laser Doppler, intravital fluorescent confocal microscopy, Western Blot analysis, and TBAR assay, respectively. The iris blood perfusion of DM and DMCUR was decreased significantly compared to CON and CONCUR (P < 0.001). Plasma glucose and HbA1c of DM and DMCUR were increased significantly compared to CON and CONCUR (P < 0.001). Leukocyte adhesion, ICAM-1, p47phox expression, and MDA levels in DM were increased significantly compared to CON, CONCUR, and DMCUR (P < 0.05). Txnip expression in DM and DMCUR was significantly higher than CON and CONCUR (P < 0.05). From Pearson's analysis, the correlation between the plasma MDA level and the endothelial functions was significant. It suggested that curcumin could ameliorate diabetic vascular inflammation by decreasing ROS overproduction, reducing leukocyte-endothelium interaction, and inhibiting ICAM-1 and NOX2 expression.

Acanthus ebracteatus Vahl. ethanol extract enhancement of the efficacy of the collagen scaffold in wound closure: a study in a full-thickness-wound mouse model.

Acanthus ebracteatus Vahl. is a Thai herb that is effective in wound healing. We sought to quantitatively determine whether or not the combined application of Acanthus ebracteatus Vahl. and a collagen scaffold will increase wound closure and angiogenesis. Balb/c mice (body weight: 22-25 g) were anesthetized with sodium thiopental. The dorsal skin incision measuring 1.5 × 1.5 cm was made and then deepened using scissors to produce a full-thickness incision down to the level of the panniculus carnosus. The size of the wound was approximately 10% of the total body surface area. The collagen sheet was implanted onto the wound. Animals were divided into 4 major groups as follows: wound with normal saline (W-NSS), wound treated with 0.3 g/kg BW of Acanthus ebracteatus Vahl. extract (W-AE (0.3 g/kg.bw)), wound implanted with collagen scaffold (W-Coll), and wound implanted with collagen scaffold and treated with 0.3 g/kg BW of Acanthus ebracteatus Vahl. (W-Coll-AE combination). On day 14, the W-Coll-AE group showed decreased wound areas and increased capillary vascularity (CV) when compared to the other 3 groups, W-NSS, W-AE0.3, and W-Coll. In the present study, the combination of AE0.3 with collagen showed the best effect on skin angiogenesis and promoted wound closure with less neutrophil infiltration.

Correlations between Endothelial Functions and ROS Detection in Diabetic Microvascular Wall: Early and Late Ascorbic Acid Supplementation.

The correlation between endothelial function and reactive oxygen species detecting from diabetic microvascular wall and the antioxidant effect of ascorbic acid (AA) during early and late phases of diabetic induction were determined. Male Spraque-Dawley rats were divided into four groups: control, diabetes rats (DM, using iv.injection of 55 mg/kg BW streptozotocin, (STZ)), and two groups of DM rats treated with AA (1 g/L, (STZ)) starting on day 2 (DM + AAday2) and week 6th (DM + AA6wk). On 12th week after STZ injection, the findings showed that in DM group, Ach (10(-5) M)-induced vasodilatation was decreased, while the number of leukocyte adhesion was increased significantly (P < 0.01). Interestingly, these abnormalities induced by DM could be protected or improved in both AA-treated groups, DM + AAday2 and DM + AA6wk. By using dihydrorhodamine 123, our findings also indicated that the existing of ROS productions on diabetic arteriolar and venular walls were different significantly (ROS(arteriole) = 165.89 ± 24.59 and ROS(venule) = 172.26 ± 34.70) (P < 0.05). Moreover by using BH4 inhibitor to induce increase in arteriolar ROS, the results also confirmed that AA could improve endothelial function with closed correlation to its potential to reduce vascular ROS content.

Effects of two modes of exercise on physical fitness and endothelial function in the elderly: exercise with a flexible stick versus Tai Chi.

OBJECTIVE: Determine the effects of exercise with flexible stick training on physical fitness and endothelial function and compare it with Tai Chi training. MATERIAL AND METHOD: Thirty older women volunteered for the present study and were divided into EF group (EF: n=16; 70.3 + 2.5 yr) and TC group (TC: n=14; 69.5 +/- 4.5 yr). Both training groups performed training assigned protocol that consisted of 70% of maximal heart rate, 40 minutes per day, four days per week for 12 weeks. Health related physical fitness and biochemical data were assessed in all participants. Post-Occlusive Reactive Hyperemia (PORH) was used to monitor endothelial function by using a Laser-Doppler fluxmeter. RESULT: The health related physical fitness was significantly higher in the EF group (p < 0.05). Plasma malondialdehyde and von Willebrand factor, an indicator of free radical damage and endothelial dysfunction, respectively as well as cholesterol level were significantly lower (p < 0.05) in the EF group. The peak Laser-Doppler flux (LDF)/baseline LDE and recovery time were significantly improved after 12 weeks of EF training (p < 0.05). This was not observed after 12 weeks of TC training. CONCLUSION: EF, a Thai novel exercise that combined endurance and strength training was a more effective exercise modality than TC for improving physical fitness and endothelial function. It improved reactive oxygen species in the elderly.

Curcumin supplementation could improve diabetes-induced endothelial dysfunction associated with decreased vascular superoxide production and PKC inhibition.

BACKGROUND: Curcumin, an Asian spice and food-coloring agent, is known for its anti-oxidant properties. We propose that curcumin can improve diabetes-induced endothelial dysfunction through superoxide reduction. METHODS: Diabetes (DM) was induced in rats by streptozotocin (STZ). Daily curcumin oral feeding was started six weeks after the STZ injection. Twelve weeks after STZ injection, mesenteric arteriolar responses were recorded in real time using intravital fluorescence videomicroscopy. Superoxide and vascular protein kinase C (PKC-ßII) were examined by hydroethidine and immunofluorescence, respectively. RESULTS: The dilatory response to acetylcholine (ACh) significantly decreased in DM arterioles as compared to control arterioles. There was no difference among groups when sodium nitroprusside (SNP) was used. ACh responses were significantly improved by both low and high doses (30 and 300 mg/kg, respectively) of curcumin supplementation. An oxygen radical-sensitive fluorescent probe, hydroethidine, was used to detect intracellular superoxide anion (O2●-) production. O2●- production was markedly increased in DM arterioles, but it was significantly reduced by supplementation of either low or high doses of curcumin. In addition, with a high dose of curcumin, diabetes-induced vascular PKC-ßII expression was diminished. CONCLUSION: Therefore, it is suggested that curcumin supplementation could improve diabetes-induced endothelial dysfunction significantly in relation to its potential to decrease superoxide production and PKC inhibition.

Curcumin suppresses gastric NF-kappaB activation and macromolecular leakage in Helicobacter pylori-infected rats.

AIM: To investigate whether curcumin could attenuate nuclear factor (NF)-kappaB p65 expression and macromolecular leakage in the gastric mucosa of Helicobacter pylori (H. pylori)-infected rats. METHODS: Twenty-five male Sprague-Dawley rats were equally divided into five groups: control rats (Control), control rats supplemented with 600 mg/kg curcumin, H. pylori-infected rats (Hp), H. pylori-infected rats supplemented with 200 mg/kg curcumin (Hp + curI), and H. pylori-infected rats supplemented with 600 mg/kg curcumin (Hp + curII). In H. pylori-infected groups, rats were inoculated with H. pylori suspension twice a day at an interval of 4 h for 3 d. Two weeks later, 200 or 600 mg/kg curcumin was given once daily to curcumin-supplemented groups for 7 d. On the day of the experiment, macromolecular leakage in gastric mucosa was examined by intravital fluorescence microscopy. The stomach tissue was removed to examine NF-kappaB p65 expression in gastric epithelial cells by immunohistochemistry. RESULTS: The expression of NF-kappaB p65 in gastric epithelial cells and the macromolecular leakage from gastric mucosal microcirculation significantly increased in the Hp group compared with the Control group. The percentages of NF-kappaB p65 immunoreactive cells in Control and Hp groups were 10.72% +/- 2.10% vs 16.02% +/- 2.98%, P = 0.004, respectively. The percentages of macromolecular leakage in Control and Hp groups were 10.69% +/- 1.43% vs 15.41% +/- 2.83%, P = 0.001, respectively. Curcumin supplementation in Hp + curI and Hp + curII groups significantly decreased NF-kappaB p65 immunoreactive cells and macromolecular leakage compared with results in the Hp group. The percentages of NF-kappaB p65 immunoreactive cells in Hp + curI and Hp + curII groups were 11.79% +/- 2.13% (P = 0.017) and 11.42% +/- 1.68% (P = 0.010), respectively. The percentages of macromolecular leakage in Hp + curI and Hp + curII groups were 12.32% +/- 2.13% (P = 0.025) and 12.14% +/- 1.86% (P = 0.018), respectively. CONCLUSION: H. pylori-induced gastric inflammation in rats is associated with increased NF-kappaB activation and macromolecular leakage which can be reduced by curcumin supplementation.

Increased NO bioavailability in aging male rats by genistein and exercise training: using 4, 5-diaminofluorescein diacetate.

BACKGROUND: Several kinds of anti-oxidants have drawn a lot of intention for their benefits on vascular protection. In addition, it has been demonstrated that exercise training could improve endothelial function by up-regulating endothelial nitric oxide synthase (eNOS) protein. Therefore, the present study aims to investigate the effects of genistein, a potent phyto-antioxidant, and exercise training on age-induced endothelial dysfunction in relation to NO bioavailability using in situ NO-sensitive fluorescent dye detection. METHODS: Male Wistar rats (20-22-month old) were divided into four groups: aged rats treated with corn oil, (Aged+Veh, n = 5), aged rats treated with genistein (Aged+Gen, n = 5, (0.25 mg/kg BW/day, s.c.)), aged rats with and without exercise training (Aged+Ex, n = 5, swimming 40 min/day, 5 days/week for 8 weeks) (Aged+Without-Ex, n = 5). Cremaster arterioles (15-35 micrometer) were visualized by fluorescein isothiocyanate labeled dextran (5 microgram/ml). The vascular response to acetylcholine (Ach; 10(-5)M, 5 ml/5 min) was accessed after 1-min norepinephrine preconstriction (10 micro molar). To determine NO bioavailability, the Krebs-Ringer buffer with 4, 5-diaminofluorescein-diacetate (3 micro molar DAF-2DA), and 10 micro- molar Ach saturated with 95%N2 and 5%CO2 were used. Changes of DAF-2T-intensities along the cremaster arterioles were analyzed by the Image Pro-Plus Software (Media Cybernatics, Inc, USA). Liver malondialdehyde (MDA) level was measured by thiobarbituric acid reaction and used as an indicator for oxidative stress. RESULTS: The results showed that means arterial blood pressure for both Aged+Gen and Aged+Ex groups were significantly reduced when compared to the Aged groups, Aged+Veh and Aged+Without-Ex (P < 0.05). Among the treated groups, Ach-induced vasodilatation were significantly increased (P < 0.05) and was associated with increased NO-associated fluorescent intensities (P < 0.05). On the other hand, MDA levels were significantly reduced (P < 0.05) when Aged+Veh was compared to Aged+Without-Ex. CONCLUSION: These findings showed that genistein and exercise training could improve age-induced endothelial dysfunction and is related to the increased NO bioavailability.

Preventive effects of genistein on leukocyte adhesion in femur venules and on bone-loss induced in ovariectomized female rats.

This study was aimed to examine effects of genistein on leukocyte adhesion in femur microcirculation in relation to bone-loss induced in ovariectomized female rats. Sixty-four female Wistar rats were divided into 4 groups: sham (daily treated with vehicle; DMSO, sc; 100 microl/day), ovariectomized rat treated with vehicle (OVX(veh)), 17beta-estradiol treated-ovariectomized rat (OVX(E2), 5 microg/kg/day, s.c.) and genistein treated-ovariectomized rat (0.25 mg/kg/day, s.c.; OVX(gen)). One and three weeks after the ovariectomy, blood flow perfusion (BF) in femur tissue was measured using laser Doppler flowmetry. Leukocyte adhesion in femur venules (15-30 microm in diameter) of each group was evaluated by intravital fluorescence videomicroscopy. The bone mineral content (BMC) was measured and expressed in terms of ratio of ash-to-dry matter weight. Serum osteocalcin and alkaline phosphatase levels were determined using chemiluminescence immunoassay. In both one and three week-OVX(veh), leukocyte adhesion increased significantly, compared to their age-matched sham groups, but it decreased significantly in OVX(gen), compared to OVX(veh) (p<0.05). In three week-OVX(gen), both BF and BMC increased significantly, but osteocalcin and alkaline phosphatase levels decreased, compared to those of three week-OVX(veh). In conclusion, genistein supplementation could effectively prevent bone-loss and microvascular endothelial dysfunction induced by estrogen deficiency.

Long-term effects of oral vitamin C supplementation on the endothelial dysfunction in the iris microvessels of diabetic rats.

The current study was aimed to investigate effects of long-term supplementation of vitamin C on the iris microcirculation in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in male Wistar-Furth rats by intravenous injection of STZ (55 mg/kg b.w.). The rats were divided into three groups: control rats (CON), STZ-induced diabetic rats (STZ), and STZ rats supplemented with vitamin C (STZ-vitC). For supplementation of vitamin C, ascorbic acid (1 g/l) was added into the drinking water. The experiments were performed at different periods (8, 12, 24 and 36 weeks) after injection of STZ. Blood glucose, tissue lipid peroxidation and plasma vitamin C were measured. To examine the endothelial function, leukocyte adhesion to the venular endothelium was evaluated in the iris post-capillaries by means of counting the number of leukocytes labeled with rhodamine 6G. Blood flow perfusion in the iris was monitored using a laser Doppler flow meter. In the STZ rats, hyperglycemia was induced with an increase in HbA(1c) and lipid peroxidation but with a decrease in the plasma vitamin C level which improved by vitamin C supplementation. The number of adherent leukocytes increased significantly, associated with reduction in the iris blood flow perfusion, at 8, 12, 24 and 36 weeks after injection of STZ. In the STZ-vitC rats, the iris blood flow perfusion was significantly increased in comparison with the STZ rats, while the leukocyte adhesion was decreased at 24 and 36 weeks. The statistical analysis shows that the leukocyte adhesion decreased with increase in the iris blood flow perfusion in STZ and STZ-vitC rats. In conclusion, vitamin supplementation suppressed leukocyte adhesion and thus endothelial dysfunction, associated with increase in iris blood flow perfusion in diabetes. The antioxidant vitamin C may be a therapeutic agent for preventing diabetic retinopathy.

Combined effects of curcumin and vitamin C to protect endothelial dysfunction in the iris tissue of STZ-induced diabetic rats.

This study was aimed to evaluate the combined effect of curcumin with vitamin C supplementation on hyperglycemic and dyslipidemia conditions and endothelial cell dysfunction induced in diabetic rats. Wistar Furth rats were used and divided into four groups: control (single injection of 0.9% sterile saline), STZ (streptozotocin, Sigma, 55 mg/kg.BW, i.v.), STZ-vitC (1 g/l ascorbic acid mixed in drinking water), STZ-cur (daily oral treatment of 300 mg/kg.BW curcumin; Cayman Chemical Co., USA), and STZ-cur+vitC (1 g/l ascorbic acid mixed in drinking water and oral treatment of 300 mg/kg.BW curcumin). On 8th week after STZ-injection, the microcirculation in the iris tissue was observed using intravital fluorescence videomicroscopy, and also leukocyte adhesion in the venule was examined for each group. Blood glucose (BG), lipid profiles, glycosylated hemoglobin (HbA1c) were measured in blood samples collected at the end of each experiment. The contents of liver malondialdehyde (MDA) were also quantified for each group. Feeding curcumin (STZ-cur) could decrease BG, HbA1c, dyslipidemia, and MDA significantly, compared to STZ. In cases of feedings curcumin with vitamin C, these results were more effective in all aspects, including leukocyte adhesion. In conclusion, curcumin might increase the effect of vitamin C in protecting the function of endothelial cells through its anti-oxidant with hypoglycemic and hypolipidemic actions.

Anti-inflammatory effects of Aloe vera on leukocyte-endothelium interaction in the gastric microcirculation of Helicobacter pylori-infected rats.

This research was aimed to investigate anti-inflammatory effects of Aloe vera on leukocyte-endothelium in the gastric microcirculation of Helicobacter pylori (H. pylori)-infected rats. Thirty-six male Sprague-Dawley rats were divided into 3 groups: control, H. pylori-infected, and A. vera-treated group (200 mg/kg b.w., twice daily). H. pylori-inoculation was induced in the rats by the administration of H. pylori solution. Intravital fluorescence videomicroscopy was used to examine leukocyte adhesion in postcapillary venules on the posterior surface of stomach area on different periods after administration of A. vera. Serum tumor necrosis factor-alpha (TNF-alpha) level was measured in blood collected at the end of experiment by using ELISA technique. The results showed that in H. pylori-infected group on day 8, the leukocyte adhesion was 13.40+/-1.00 cells/100 microm vessel length and the TNF-alpha was 76.76+/-23.18 pg/ml, which increased significantly (p < 0.05), compared with the control group (leukocyte adhesion(control) = 2.54+/-0.6 cells/100 microm vessel length and TNF-alpha(control) = 9.92+/-2.62 pg/ml). Treatment with A. vera reduced the leukocyte adhesion (5.5+/-0.5 cells/100 microm vessel length), and TNF-alpha (26.31+/-6.38 pg/ml) significantly (p < 0.05). In conclusion, H. pylori enhanced leukocyte-endothelium interaction in the posterior stomach area markedly. This enhancement in leukocyte-endothelium interaction could be improved by the treatment of A. vera, associated with reduction in TNF-alpha level.

Effects of Aloe vera and sucralfate on gastric microcirculatory changes, cytokine levels and gastric ulcer healing in rats.

AIM: To compare the effects of Aloe vera and sucralfate on gastric microcirculatory changes, cytokine levels and gastric ulcer healing. METHODS: Male Spraque-Dawley rats (n=48) were divided into four groups. Group1 served as control group, group 2 as gastric ulcer group without treatment, groups 3 and 4 as gastric ulcer treatment groups with sucralfate and Aloe vera. The rats from each group were divided into 2 subgroups for study of leukocyte adherence, TNF-alpha and IL-10 levels and gastric ulcer healing on days 1 and 8 after induction of gastric ulcer by 20% acetic acid. RESULTS: On day 1 after induction of gastric ulcer, the leukocyte adherence in postcapillary venule was significantly (P<0.05) increased in the ulcer groups when compared to the control group. The level of TNF-alpha was elevated and the level of IL-10 was reduced. In the ulcer groups treated with sucralfate and Aloe vera, leukocyte adherence was reduced in postcapillary venule. The level of IL-10 was elevated, but the level of TNF-alpha had no significant difference. On day 8, the leukocyte adherence in postcapillary venule and the level of TNF-alpha were still increased and the level of IL-10 was reduced in the ulcer group without treatment. The ulcer treated with sucralfate and Aloe vera had lower leukocyte adherence in postcapillary venule and TNF-alpha level. The level of IL-10 was still elevated compared to the ulcer group without treatment. Furthermore, histopathological examination of stomach on days 1 and 8 after induction of gastric ulcer showed that gastric tissue was damaged with inflammation. In the ulcer groups treated with sucralfate and Aloe vera on days 1 and 8, gastric inflammation was reduced, epithelial cell proliferation was enhanced and gastric glands became elongated. The ulcer sizes were also reduced compared to the ulcer group without treatment. CONCLUSION: Administration of 20% acetic acid can induce gastric inflammation, increase leukocyte adherence in postcapillary venule and TNF-alpha level and reduce IL-10 level. Aloe vera treatment can reduce leukocyte adherence and TNF-alpha level, elevate IL-10 level and promote gastric ulcer healing.

Asian traditional medicine (ATM): recent progress based on scientific evidences.

Asian traditional medicine (ATM) (herbal medicine, acupuncture or moxibution) has gained some popularity among communities in Asia. Some therapies employed in ATM have been verified using modern techniques, but the significance of ATM has still remained unclear. This symposium was focused on experimental data obtained recently.

Genistein replacement therapy on endothelial dysfunction and bone loss in bilateral ovariectomized rats.

The objective of this study is to examine roles of genistein in postmenopausal induced-endothelial dysfunction and bone loss, the ovariectomized (OVX) rat model was used. The animals were divided into three groups of sham treated with vehicle (DMSO 100 microl/day; Sham(veh)), OVX treated with vehicle (OVX(veh)) and OVX treated with genistein (0.25 mg/kg/day; OVX(gen)). At 3 and 7 weeks after the surgery, endothelial dysfunction in mesenteric microcirculation of each group was determined by using intravital fluorescence microscopy and analyzed with digital image software. The parameters of bone mass density (BMD) and bone formation marker were represented by percentage of ash/dry matter and osteocalcin activity (using radioimmunoassay (RIA)), respectively. Mean arterial pressures (MAP) in OVX(veh) groups were significantly increased compared to their aged-matched sham groups (p<0.05). Interestingly, the treatment of genistein could significantly attenuate this abnormality (p<0.001). Besides, it could increase the vascular response to acetylcholine (Ach; 10(-6) M) significantly compared to OVX-rats (p<0.05). Moreover, BMD and osteocalcin activity were significantly increased in Ovx(gen) as well. Therefore, our findings suggested that genistein supplementation could effectively prevent endothelial dysfunction and bone loss in OVX-rat model.

Vitamin C supplementation could reverse diabetes-induced endothelial cell dysfunction in mesenteric microcirculation in STZ-rats.

This paper was aimed to study whether vitamin C supplementation reverses the diabetes-induced endothelial cell dysfunction occurred in streptozotocin (STZ)-rats or not. The animals were divided into four groups: control and diabetes rats (DM, using iv. injection of 50 mg/kg.bw STZ), and two DM rats treated with vitamin C (1 g/l) starting on day 2 (DM + VitC(day2)) and week 6th after STZ-injection (DM + VitC(6wks)). The mesenteric microcirculation was observed using fluorescence videomicroscopy. Based on the recorded videoimages, microvascular responses to acetylcholine (Ach; 10-5 M) and number densities of leukocyte adhesion in venules were evaluated using the Global Lab II image software. In DM group, blood glucose and glycosylated hemoglobin were significantly increased, while the body weight and plasma vitamin C levels were decreased significantly compared to their controls. Ach-induced vasodilation was decreased, while the number of leukocyte adhesion was increased significantly compared to their controls (p<0.01). These abnormalities induced by DM were prevented by supplementation of vitamin C in DM + VitC(day2) group. Six-weeks delayed treatment of vitamin C (DM + VitC(6wks)) demonstrated increase in the Ach-induced vasodilation with significant decrease in the leukocyte adhesion. It was indicated that vitamin C supplementation could reverse diabetes-induced endothelial cell dysfunction in mesenteric microcirculation.

Ganoderma lucidum suppresses endothelial cell cytotoxicity and proteinuria in persistent proteinuric focal segmental glomerulosclerosis (FSGS) nephrosis.

A persistent proteinuria is commonly observed in nephrotic patient with focal segmental glomerulosclerosis (FSGS) under treatment with prednisolone+/-cyclophosphamide or with vasodilators (ACEI+AII receptor antagonist, calcium channel blocker and antiplatelet agent). Fourteen such patients with persistent proteinuria were subject to be treated with Ganoderma lucidum. Initial study revealed an enhanced endothelial cell cytotoxicity induced by patient's serum, and an altered immunocirculatory balance with predominant proinflammatory cytokine TNF alpha activity in the presence of defective anti-inflammatory cytokine interleukin-10. Treatment with Ganoderma lucidum suppressed endothelial cell cytotoxicity, restored immunocirculatory balance and successfully suppressed proteinuria in all of these 14 patients.

Asian traditional medicine (ATM) based on in vivo microcirculation evidence.

Asian traditional medicine (ATM) (herbal medicine, acupuncture or moxibution) has gained some popularity among communities in Asia, but there are still few evidences to prove the effectiveness of such therapeutic measures. A symposium was held with aim at the effectiveness of Asian traditional therapies in views of in vivo microcirculation. This report is concerned with the symposium, including Asian activities for ATM.