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Objective To evaluate the inhibitory effect of chidamide combined with curcumin on the proliferation of cutaneous T-cell lymphoma (CTCL) cell line Hut78,as well as their promotive effect on its apoptosis,and to explore their therapeutic mechanisms in CTCL.Methods Some Hut78 cells were treated with different concentrations of chidamide (0.3,0.6,1.2,2.4 μmol/L) and 10 μ mol/L curcumin alone or the combination of 1.2 μ mol/L chidamide and 10 μ mol/L curcumin for 24,48 and 72 hours separately.MTS assay was conducted to estimate cell viability at each time point.After selection of chidamide concentrations,some Hut78 cells were treated with chidamide (0.6 and 1.2 μmol/L)and curcumin (10 μmol/L) alone or in combination (1.2 μmol/L chidamide and 10 μmol/L curcumin) for 24 hours,then,flow cytometry was performed to detect cell apoptosis and analyze cell cycle,real-time (RT)-PCR and Western-blot analysis were conducted to quantify the mRNA and protein expressions of apoptosis-associated genes Fas,caspase 8,nuclear factor (NF)-κB p65 as well as cell cycle-associated genes P21,CDK2 and cyclin E respectively.Statistical analysis was carried out by repeated-measures analysis of variance,one-way analysis of variance and the least significant difference (LSD)-t test.Results Chidamide could significantly inhibit the proliferation of Hut78 cells in a dosedependent and time-dependent manner (F =266.558,564.966,respectively,both P ＜ 0.001).After 48-and 72-hour culture,the combination of 1.2 μmol/L chidamide and 10 μmol/L curcumin showed significantly stronger inhibitory effect on cell proliferation compared with 1.2 μmol/L chidamide or 10 μmol/L curcumin alone (all P ＜ 0.001).As flow cytometry showed,the percentage of apoptotic cells was significantly higher in the combined treatment group than in the 0.6-,1.2-μmol/L chidamide groups and 10-μmol/L curcumin group (all P ＜ 0.001).Compared with the 0.6-μmol/L chidamide group and 10-μmol/L curcumin group,the combined treatment group and 1.2-μmol/L ehidamide group both showed significantly increased proportion of cells at G0/G1 phase and mRNA expressions of Fas,caspase 8 and P21,but decreased proportion of cells at S phase or G2/M phase and mRNA expressions of NF-κB p65,CDK2 and cyclin E (P ＜0.05 for proportion of cells at different phases,P ＜ 0.001 for mRNA expressions of different genes).Furthermore,the mRNA expression of Fas was significantly higher in the combined treatment group than in the 1.2-μmol/L chidamide group (P ＜ 0.001),while no significant differences were observed in the mRNA expressions of caspase 8,P21,NF-κB p65,CDK2 and cyclin E or the proportion of cells at any phase between the combined treatment group and 1.2-μmol/L chidamide group (all P ＞ 0.05).Western-blot analysis showed that protein expressions of Fas,caspase 8 and P21 significantly increased,but those of NF-κB p65,CDK2 and cyclin E significantly decreased in the combined treatment group compared with the 0.6-,1.2-μmol/L chidamide groups and blank control group receiving no treatment,which were in accordance with the above changes in mRNA expressions of these genes.Conclusion Chidamide can inhibit the growth of the CTCL cell line Hut78 by directly decelerating cell proliferation and inducing cell apoptosis,and the combibation with curcumin can markedly enhance the inhibitory effect of chidamide on the growth of Hut78 cells.
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Objective To estimate the value of detection of Merkel cell carcinoma polyomavirus (MCPyV)in the diagnosis of Merkel cell carcinoma (MCC).Methods Two cases of MCC were studied using light microscopy and immunohistochemistry.PCR was performed to detect DNA sequences encoding MCPyV large T antigen(LT)and viral protein 1 (VP1) in paraffin-embedded tissue specimens from the two patients with MCC,five patients with T cell lymphoma,two normal human controls,as well as in two T cell lymphoma cell lines MAC 1 and MAC2.DNA sequencing was also carried out.Results Both of the patients with MCC were male.The patient 1 presented with a mass in the right anterior shin for more than one year,and the patient 2 had a mass in the left knee for more than six months.Skin examination revealed densely distributed pink nodules in the right anterior shin,with confluence into an indurated plaque which measured 10 cm × 8 cm with superficial erosion,exudates and crusts and was surrounded by multiple irregularly sized erythematous nodules with limited mobility in the patient 1,as well as a royal blue,hard,poorly marginated nodular mass measuring 5 cm × 4 cm in the left medial knee with limited mobility in the patient 2.Pathological manifestations were similar in the two patients.Tumor cells were uniform with large hyperchromatic nuclei,eosinophilic and sparse cytoplasm,and fine chromatin.Mitotic figures were easily seen.Immunohistochemistry revealed that the tumor cells stained positive for pan-cytokeratin,synuclein (Syn),neuron-specific enolase (NSE),chromogranin (CgA),CK20,and Ki-67 (＞ or =60％),but negative for S100 protein,HMB45,CD34,thyroid transcription factor 1 (TTT-1),CK7 and leukocyte common antigen (LCA).MCPyV DNA was detected in both MCC specimens,but absent in the other skin specimens or T cell lymphoma cell lines.Conclusions MCC has distinctive clinical and pathological appearance.Immunohistochemistry and detection of MCPyV DNA sequences using PCR may be beneficial to the definitive diagnosis of MCC.
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Objective To explore the expression and significance of Annexin Ⅱ protein in ovarian adenocar-einoma.Methods The expressions of Annexin Ⅱ protein were detected in 48 cases of ovarian adenocarcinoma and 10 cases of ovarian adenoma by SP immunohistochemistry.Results The positive rates of expression of Annexin Ⅱprotein in ovarian adenoma were 50.00% (5/10), and were 81.25% (39/48) in ovarian adenocareinoma (χ2 =4.414, P <0.05), in well and moderately-differentiated, poorly differentiated ovarian serous adenocarcinoma were 93.33% (14/15) and 78.95% (15/19) respectively (χ2 = 1.383 ,P0.05 ), and on the Ⅰ and Ⅱ stage, Ⅲ and Ⅳ stage of ovarian serous adenocarcinoma were 60.00% (6/10) and 95.83% ( 23/24 ) respectively (χ2=7.226, P <0.05).The positive rate in the group of ovarian serous adenocarcinoma with lymph node metastasis was 90.00% (27/30), while 50.00% (2/4) in the group without lymph node metastasis (χ2=4.502, P<0.05).Conclusion The positive expression of Annexin Ⅱ protein may be correlated with the carcinogenesis, development and lymph node metastasis of ovarian serous adenocarcinoma.
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Objective:To probe the different influence of pericardial devascularization by preserving vagus trunk(VTPPD) and pericardial devascularization (PD) on portal hypertensive gastropathy (PHG).Methods:77 patients with portal hypertension were divided into VTPPD and PD group,the VTPPD group included 36 cases,and PD group included 41 cases.Varices of esophagus and fundus of stomach and PHG were observed by gastroscopy before and 3 weeks after operation in all cases,and compared postoperative incidence of PHG in the 2 groups.Results:In all cases,Varices of esophagus and fundus of stomach disappeared or relieved obviously.The incidence of PHG in VTPPD group before operation was 55.6%(20/36),and that after operation was 69.4%(25/36),the former was not higher statistically(P=0.224);the incidence of PHG in PD group before operation was 61.0%(25/41),and that after operation was 87.8%(36/41),the former was not higher than the latter statistically(P=0.005);and the postoperative incidence of PHG in PD group was higher significantly than that in VTPPD group (P=0.048).There were 8(22.2%,8/36)patients whose degree of PHG aggravated in VTPPD group,and there were 19(46.3%,19/41)patients whose degree of PHG aggravated in PD group,the rate of the former was significantly lower than that of the latter(P=0.027).Conclusion:Comparing with the classic portoazygous devascularization,VTPPD can reduce the incidence and the degree of PHG.