Objective To observe the effect of bcl-2 antisense oligodexyonucleotide (ASODN) by phosphorothiote-modification on the expression of bcl-2 mRNA and protein in human melanoma A375 cells line. Methods With the phosphorothiote-modification and liposome-encapsulation of ASODN, A375 cells were divided into ASODN group, nonsense oligodexyonucleotide (SODN) group and control group. Bcl-2 mRNA and protein were detected by RT-PCR and immunohistochemistry, respectively. Results The expression of bcl-2 protein was remarkably decreased in ASODN group than that in SODN group and control group (53.14 ±4.26 vs 138.22 ± 8.45, 53.14 ± 4.26 vs 141.08 ± 7.83, both P ＜ 0.01 ). The level of bcl-2 mRNA was significantly lower than that in SODN group and control group (0.38 ± 0. 11 vs 0.96 ±0.13, 0. 38 ± 0.11 vs 0. 97 ± 0. 14, both P ＜ 0. 01 ). Conclusion Bcl-2 antisense oligodexyonucleotide could down-regulate the bcl-2 level and block its protein expression.

Objective To investigate expressions of STAT3 and phosphorylated STAT3(p-STAT3)in cutaneous malignant melanoma(CMM)lesions, and to explore their significance. Methods The expressions of STAT3 and p-STAT3 were detected with an immunohistochemical method using polymer enzyme-labeled secondary antibodies in skin specimens from 22 CMM lesions and 23 pigmented nevus lesions. The effects of STAT3 and p-STAT3 expressions on lymph node metastasis of CMM were evaluated. Results The expression rate of STAT3 was significantly higher in CMM lesions than in pigmented nevus lesions (95.5% vs. 56.5%, χ2 = 9.23, P < 0.05), so was that of p-STAT3 (90.9% vs. 43.5%, χ2 =11.38, P < 0.05). STAT3 expression in CMM lesions had no obvious effects on lymph node metastasis of CMM (P > 0.05), while the elevated expression of p-STAT3 in CMM lesions was a predictive factor for lymph node metastasis in CMM (OR = 1.88, 95% CI: 1.05 - 3.38, P < 0.05). Conclusion Increased expression of STAT3 and p-STAT3 in CMM may play important roles in the pathogenesis of CMM, and elevated p-STAT3 expression may contribute to invasion and metastasis of CMM.

Objective To investigate the effects of all -trans retinoic acid (ATRA),acitretin and tazarotene on apoptosis and active caspase-3 of human melanoma cells A375 and to discuss the relevant significance.Methods The effects of retinoids on apoptosis and active caspase-3 of A375 cells were examined in vitro.Early apoptosis analysis by double staining with annexin V-FITC and PI,and active caspase 3 analysis by the staining of FITC conjugated monoclonal rabbit anti active caspase-3 antibody were measured by flow cytometer.Results ATRA and acitretin induced apoptosis of A375 cells (both P＜0.05),but tazarotene was not effective (P＞0.05).ATRA and acitretin elevated the cell population with detectable active caspase-3 (both P＜0.05),but tazarotene was not effective either (P＞0.05).Acitretin played a most prominent role among the retinoids. Early apoptosis ratio was significantly and positively correlated with active caspase-3 percentage in both ATRA and acitretin (P＜0.05).Conclusions Acitretin and ATRA induce apoptosis of A375 cells in which caspase is critical.Retinoids are prospectively applied as an assisting alternative for treating melanoma.

Objective To explore the mechanism and inhibition of botulinum toxin type A (BTXA) on hypertrohic scar fibroblasts.Methods The cells were treated by 0 (control),0.2,0.4,0.8 U/ml BTXA for 48 h.Cell viability was detected by MTT assay.Cell apoptosis was detected by Hoechst staining.Cell cycle was detected by flow cytometry.The level of cell cycle related protein D1 (Cyclin D1),proliferation nuclear antigen (PCNA) and activation of phosphatidylinositol 3 kinase (PI3K) / protein kinase B (AKT) signaling pathway were assayed by western blot.Results Compared with control group(0.75±0.07),0.2,0.4,0.8 U/mL BTXA(0.59 ± 0.06,0.43 ± 0.04,0.34± 0.03) inhibited hypertrohic scar fibroblasts cell viability,increased cell apoptotic rate[control group(2.38±0.24)％;BTXA(15.79±1.54)％,(27.32±2.69)％,(38.46±3.90)％],down-regulated the expression of Cyclin D1(control group 1.57±0.18;BTXA 0.93±0.07,0.42±0.04,0.35±0.03) and PCNA(control group 1.46±0.16;BTXA 0.50±0.05,0.59±0.05,0.37±0.03),inhibited the expression of PI3K(control group 0.98±0.06;BTXA 0.49±0.04,0.50±0.04,0.39±0.03) and the phosphorylation of AKT(control group 1.38±0.08;BTXA 0.97±0.06,0.60±0.04,0.29± 0.02),made cell cycle arrested in G1 phase,The difference was statistically significant (P＜0.05).Conclusion These results suggested BTXA inhibit proliferation via blocking the activation of PI3K/AKT signal pathway and down-stream related cell cycle related protein.

Objective Analysis and adscription of volatiles from Guizhi Tang and study on its improvement of the learning and memory in dementia mice induced by scopolamine.Methods The volatile oil from Guizhi Tang(GZT),Guizhi and Shengjiang was extracted using steam distillation method and was analyzed by GC-MS. Morris water maze and step-down test were carried out for obtain the difference of the learning and memory improvement in 40 ICR mice from randomized groups, such as the control group, the model group, the donepezil group (2 mg/kg), the low dose of volatile oil of GZT (5 mg/kg), and the high dose of volatile oil of GZT (20 mg/kg), and ACh, AchE, BchE and chE in serum were detected by ELISA. Results Among 38 identyfied volatile ingredients from GZT, 18(44% in weight) was from Guizhi, and 9 was from Shengjiang. Compared with the model group, the low and high dose of GZT volatile oil significantly increased swimming distance ratio in destination quadrant (26.74% ± 16.42%vs.9.42% ± 8.50%, P<0.05); goal quadrant time scale (43.51% ± 25.12%vs. 14.50% ± 12.23%,P<0.05)) increased significantly than the model group ; the number of errors in the experiment platform (1.63 ± 1.19vs. 0.25 ± 0.46, P<0.05) obviously increased than model group ; platform test in the made errors times (0.57 ± 0.98vs. 4.43 ± 2.4, P<0.05) significantly reduced. The GZT total volatile oil groups significantly reduced cognitive obstacles small rat serum in the cholinester enzyme (chE) (140.90 ± 3.27, 144.79 ± 6.71vs. 134.49 ± 3.36,P<0.05); acetylcholinesterase (AchE) (3.30 ± 1.31, 3.94 ± 0.78 vs.8.52 ± 3.39,P<0.05); butyrylcholinesterase (BchE) (3.22 ± 0.45, 3.66 ± 0.53vs. 7.99 ± 0.79,P<0.05); and acetylcholine (Ach) (4.10 ± 0.38, 3.03 ± 0.25vs.1.72 ± 0.50, P<0.05) significantly increased.Conclusions The GZT volatile oil mainly from Guizhi and Shengjiang can improve the learning and memory ability in dementia mice induced by scopolamine via a cholinergic mechnism.

To investigate the effects of ATRA, acitretin and tazarotene on the growth and apoptosis of human tongue squamous cell carcinoma cell line Tca8113. The effect of retinoids on growth of Tca8113 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis with double staining with Annexin V-FITC and PI, and active caspase-3 analysis with the staining of FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. Streptavidin-biotin complex (SABC) immunocytochemical assays were employed for the detections of Bax/Bcl-2 proteins expressions. Our results showed that the retinoids inhibited growth of Tca8113 cells in a dose- and time-dependent manner with maximal inhibition 24 h after treatment of 10-5 mol/L. 10-5 mol/L retinoids altered cell cycle distribution of Tca8113 cells, revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. 10-5 mol/L retinoids significantly induced apoptosis of Tca8113 cells (all P<0.05), elevated the cells population with detectable active caspase-3 (P<0.05 for all), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (all P<0.05). Acitretin played a most prominent role among the retinoids. It is concluded that the inhibition of cell cycle progress of Tca8113 cells by ATRA, acitretin and tazarotene is one of the possible mechanisms for proliferation arrest of Tca8113 cells elicited by the retinoids. The retinoids mediate apoptosis in Tca8113 cells that may be caspase-dependent through mitochondria pathway. High concentration retinoids inhibit growth of Tca8113 cells in vitro by interfering with proliferation and inducing apoptosis of cells. Acitretin may be an alternative medicine for the prevention and treatment of tongue squamous cell carcinoma.

Objective:To investigate clinical manifestations, pathological features and diagnosis of eczematoid clear cell acanthoma of the nipple/areola.Methods:The clinical manifestations, histopathological features, special staining results and immunohistochemical features of a case of eczemtoid clear cell acanthoma of the nipple/areola firstly reported in China were analyzed, and compared with those of similar cases in foreign literature.Results:The female patient presented with recurrent pruritic rashes on the left nipple and areola for over 2 years. Skin examination showed hypertrophic skin on the left nipple and areola, and scattered erythema, hypopigmented macules and hyperpigmented macules on the areola, which were covered with a few crusts and scales. Histopathological examination of the skin lesions showed focal epidermal crusts and scales, focal parakeratosis, extended and fused rete ridges, thickened spinous layer, focal spongiosis, clear cell clumps in the spinous cell layer, telangiectasia in the superficial dermis, with infiltration of a few eosinophils and neutrophils. Periodic acid-Schiff staining showed positive results, and immunohistochemical staining revealed positive reaction for epithelial membrane antigen. Topical treatment with triamcinolone acetonide and econazole cream was effective, and topical application of 3% boric acid solution could alleviate exudation. During the 6-month follow-up, the patient experienced intermittent recurrence twice, and responded well to the above treatment.Conclusions:Eczematoid clear cell acanthoma of the nipple/areola has unique clinical and pathological features, revealing that it′s a new subtype of clear cell acanthoma. Pathological examination is the gold standard for its diagnosis.