ObjectiveTo compare quantitative proteomic analysis of bromotetrandrine (W198) which was a Class Ⅰ new antitumor drug in China and tetrandrine (Tet) in K562 cell line using 18O-labeling method.MethodsTo illustrate its mechanism,a shotgun quantitative proteomic strategy employing 2D LC-MS-MS and trypsin catalyzed 18O-labeling quantification was carried out in this study.Compared to normal chronic leukemia cell line K562 and K562 induced by Tet,the proteomic changes of K562 induced by W198 were investigated.In order to validate the quantitation by the 18O-labeling,the analysis was done on an equivalent sample composed of the same amount of labeled and unlabeled proteins from normally cultured cells to act as a reference to the comparative sample.ResultsA threshold of ± 2-fold change for deciding whether a protein concentration was changed was settled for the following experiments.Comparing the 105 identified soluble proteins' expression levels of the apoptosis starting up K562 cells after W198 induction with the normally cultured cells,16 proteins were found with significantly altered expression levels after W198 treatment.Eight proteins were up-expressed including HMGB2,peroxiredoxin-2,and eIF4A-I,etc.Eight proteins were down-expressed including TCP-1,GRP94,GST-π,and SFGHs,etc.Compared to K562 induced by Tet,eight proteins of K562 were found with significantly altered expression levels after W198 treatment.Five proteins were up-expressed including HSP 90-β and 40S ribosomal protein S15a,etc.Three proteins were down-expressed including phosphoglycerate kinase 1,isoform 5 of interleukin enhancer-binding factor 3,etc.ConclusionThe 18O-labeling MS-MS-based method is ideal as a discovery tool,but it is not suitable for validation using a large number of samples.Other more effective methods,such as Western blotting should be used for further validation of candidate cancer proteins discovered from 18O-labeling samples.In total,105 soluble proteins were discovered,and 16 proteins were found with significantly altered expression levels after W198 treatment.These repressed or activated proteins are the potential drug targets of W198,which may provide novel targets for future development ofbiomarkers for cancer therapy.

[ABSTRACT]OBJECTIVETo evaluate the effectiveness of low temperature radiofrequency ablation on tongue cancer in early stage (<2 cm T1 stage) .METHODS High differentiated tongue squamous cell carcinoma(<2 cm T1 stage) were removed with radiofrequency ablation in 11 patients and with high-frequency electrotome in 20 patients from 2009 to 2014 in our hospital. All the patients underwent elective neck dissection(I,Ⅱ,Ⅲ regions). Intraoperative blood loss, VAS ratings of post-operative pain, post-operative bleeding rate and the recurrence rate of tongue cancer or lymph node metastasis were compared between the two groups.RESULTSIn the radiofrequency ablation group, the mean intraoperative blood loss was 13.82±7.40ml, the VAS ratings of post-operative pain were 3.8±1.3 (day 1), 2.5±0.7 (day 3) and 1.8±0.6(day 5), post-operation bleeding occurred in one case, and lymph node metastasis occurred in one case at 6 month after operation. In the high-frequency electrotome group, the mean intraoperative blood loss was 40.55±12.03ml, the VAS ratings of post-operative pain were 6.8±1.3(day 1), 4.4±1.1(day 3) and 2.3±0.7(day 5), post-operation bleeding occurred in one case, and lymph node metastasis occurred in 3 cases at 6 month to one year after operation. The intraoperative blood loss and post-operative pain in radiofrequency ablation group were significantly lower and less than that in the high-frequency electrotome group.CONCLUSIONRadiofrequency ablation is a promising method for early stage tongue cancer with less blood loss, invasiveness and complications.

【Objective】To observe the effect of Huangqi Sijunzi Decoction(HSD) combined with parenteral nutrition in promoting postoperative recovery of gastric cancer patients.【Methods】Eighty gastric cancer patients were randomized into two groups.On the 3rd day after operation,the two groups received parenteral nutrition,and the treatment group(group A) was given HSD additionally.The anal exsufflation time,incision infection rate,biochemical indexes and immune indexes of the two groups were observed.【Results】The anal exsufflation time was(2.40?0.33) days in the treatment group and(3.82?0.26) days in the control group,the difference being significant(P0.05).The increase of pre-albumin in the treatment group was obvious as compared with that in the control group(P0.05).The increase of immune indexes was not obvious in the control group(P

Aim To explore the effect of naloxone on th e plasma level of ?-endorphin (?-EP) in rats with alcoholic fatty liver (AFL) and its possible mechanism. Meth ods Forty-eight Wistar male rats were divided randomly into four grou ps: Model group (n 12), low doses treatment group (n12), high doses tre atment group (n12) and normal control group (n12). By the end of the f ourth week, all the rats were weighted, narcosised, sacrificed, and got the whol e liver. The level of ?-EP in plasma and the level of superoxidized dismutase (SOD) and malondialdehyde (MDA) in plasma and liver tissue were measured.The liv er samples were analyzed for histopathology with microscope. Results Rat models of AFL were established successfully. The level of plasma SOD i ncreased while MDA and ?-EP decreased in low doses treatment group and high do ses treatment group markedly contrast with model group (P

Objective To investigate the methylation status of 14-3-3σ promoter in nasopharyngeal carcinoma cell lines and the influence of de-methylation treatment on 14-3-3σ expression. Methods Methylation status of 14-3-3σ gene promoter and 14-3-3σ mRNA expression were detected by methylation specific PCR (MSP) and RT-PCR in nasopharyngeal carcinoma cell lines CNE1, CNE2,5-8F,6-10B and immortalized nonneoplastic human nasopharyngeal epithelial cell line, NP69. Four nasopharyngeal carcinoma cell lines were treated with 5-asa-2' -deoxycytidine(5-aza-2dC) in different concentration for 72 h, then 14-3-3σ promoter meth-ylation status and m RNA expression were assessed, and western-blot was performed to detect the expression of 14-3-3σ protein. Results 14-3-3σ promoter methylation was detected by MSP in all of the four nasopharyn-geal carcinoma cell lines untreated by 5-aza-2dC whereas not in the treated ones or the immortalized human na-sopharyngeal epithelial cell line, NP69. Accordingly, 14-3-3σ mRNA expression was significantly discounted in untreated nasopharyngeal carcinoma cell lines as compared with NP69. 5-aza-2dC treatment dose-depend-ently reversed 14-3-3σ promoter methylation status and consequently upregulated the expression of 14-3-3σmRNA and protein in 4 nasopharyngeal carcinoma cell lines. High-differentiated CNE1 was more sensitive to 5-aza-2dC than lowly-differentiated CNE2, 5-8F and 6-10B. Conclusion Promoter methylation directly leads to decreased 14-3-3σ gene expression in nasopharyngeal carcinoma cell lines, and 14-3-3σ promoter de-methylation perhaps indicates a new target for nasopharyngeal carcinoma treatment.

Electric Stimulation ; Facial Nerve ; physiopathology ; Facial Paralysis ; therapy ; Humans

Objective:To sutdy the effects of the polysaccharide(S3) separated out in 60% alcohol aqueous from OCA on their immunomodulatory functions in mice.Methods:After screening the most effective component of S3 by phagocytic function of macrophage,further tests about the effects both on the humoral immunity and the cellular immunity of S3 in mice were studied.Results:S3 simulated the production of the antibody on the haemolysin test,selectively enhanced ConA induced murine proliferation of lymphocytes(P

Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.

Objective:To observe the expression of Ki-67,P53 and LAT1 in tissues of esophageal squamous cell carcinoma and precancerous lesions and to investigate its clinical significance.Methods: Immunohistochemical method was used to detect the expressions of Ki-67,P53 and LAT1 in tissues from 20 cases of normal esophageal mucosa,68 cases of precancerous lesions including 21 cases of mild atypical hyperplasia,22 cases of atypical hyperplasia,25 cases of severe atypical hyperplasia and cancer tissues from 34 esophageal cancer patients.The relationship between the expression of the three in esophageal carcinoma was analysed.Results: Re-spectively,the positive expression rate of Ki-67 in normal esophageal mucosa, mild atypical hyperplasia, moderate dysplasia, severe dysplasia and carcinoma was 0%( 0/20 ) , 23.8%( 5/21 ) , 40.9%( 9/22 ) , 76.0%( 19/25 ) , 82.4%( 28/34 ) and the positive expression rate of P53 was 0%(0/20),14.3%(3/21),31.8%(7/22),48.0%(12/25),67.6%(23/34) and the positive expression rate of LAT1 was 0%( 0/20 ) , 19.0%( 4/21 ) , 36.4%( 8/22 ) , 52.0%( 13/25 ) , 76.5%( 26/34 ).The rank correlation analysis showed that the positive expression of Ki-67,P53 and LAT1 were significantly correlated with histological grade(r=0.626,0.427, 0.586,P<0.01) and the expression of Ki-67,P53 was positively correlated with LAT1 in esophageal carcinoma tissues(r=0.428, 0.596,P<0.01).Conclusion:Abnormal expression of Ki-67,P53,LAT1 protein was significant related to carcinogenesis of esophageal cancer and the combined detection of the three has important clinical significance.

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.