Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

Objective:To compare the clinical efficacy of Du's heat-reinforcing method and Western medication in treating diarrhea-predominant irritable bowel syndrome (IBS-D).Methods:Sixty-five IBS-D patients were randomized into two groups by the random number table.Thirty-three cases in the treatment group were intervened by acupuncture with Du's heat-reinforcing method,while thirty-two cases in the control group were given oral administration of pinaverium bromide tablets.The intervention lasted 4 weeks for both groups.The IBS symptom severity score (IBS-SSS),IBS-quality of life (IBS-QOL) and hospital anxiety and depression scale (HAD) were adopted to evaluate the therapeutic efficacy.Results:The two groups each had two dropout cases.The clinical recovery rate was 38.7％ and the total effective rate was 90.3％ in the treatment group versus 13.3％ and 66.7％ in the control group.The treatment group was superior to the control group in both clinical recovery rate and total effective rate (both P＜0.05).After 1-week and 4-week treatment,respectively,the IBS-SSS scores were lower compared with the baseline in both groups,and the intra-group differences were statistically significant (both P＜0.05);the score in the treatment group was lower than that in the control group,and the between-group difference was statistically significant (both P＜0.05).After 4-week treatment,the component scores of IBS-QOL showed improvements in both groups,and the treatment group was superior to the control group in the improvements of dysphoria,interference with activity,health worry and food avoidance (all P＜0.05).The anxiety and depression scales of HAD (HAD-a,HAD-d) in the treatment group and the HAD-a in the control group obtained significant improvements (all P＜0.05);the scores of HAD-a and HAD-d in the treatment group were significantly better than those in the control group (both P＜0.05).Conclusion:Acupuncture with Du's heat-reinforcing method can effectively ease the symptoms of IBS-D,improve the quality of life and the state of anxiety and depression.It can produce a more significant efficacy than oral administration of pinaverium bromide tablets.

AIM: To study the pharmacodynamic effect of Chanfuan Oral Solution. METHODS: Acute blood stagnation caused by adrenaline, ankle swelling, ear edema and writhing caused by acetic acid were taken for experiment models. RESULTS: Chanfuan Oral Solution could significantly reduce blood viscosity. Results also showed that it had inhibitory effect on rat's ankle swelling, ear edema and reduction in ratio of writhing of mice. CONCLUSION: These results suggest Chanfuan Oral Solution is an effective drug for reducing blood viscosity, anti inflammation and analgesia.

Acute Disease ; Adenoviridae Infections ; epidemiology ; Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Respiratory Tract Infections ; epidemiology ; virology

Adolescent ; Adult ; Anemia, Aplastic ; blood ; Case-Control Studies ; Female ; Humans ; Isoenzymes ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Protein Kinase C ; metabolism ; Protein Kinase C-theta ; T-Lymphocytes, Cytotoxic ; cytology ; Th1 Cells ; cytology ; Young Adult

Molecular target-based cancer therapy is playing a more and more important role in cancer therapy because of its high specificity, good tolerance and so on. There are different kinds of molecular targeted drugs such as monoclonal antibodies and small molecular kinase inhibitors, and more than 50 drugs have been approved since 1997. When the first monoclonal antibody, rituximab, was on the market. The development of molecular target-based cancer therapeutics has become the main approach. Based on this, we summarized the drugs approved by FDA and introduced their mechanism of actions and clinical applications. In order to incorporate most molecular targeted drugs and describe clearly various characteristics, we divided them into four categories: drugs related to EGFR, drugs related to antiangiogenesis, drugs related to specific antigen and other targeted drugs. The purpose of this review is to provide a current status of this field and discover the main problems in the molecular targeted therapy.
Angiogenesis Inhibitors ; Antibodies, Monoclonal ; Drug Delivery Systems ; Humans ; Molecular Targeted Therapy ; Neoplasms ; drug therapy ; Protein Kinase Inhibitors ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors

Objective To investigate the effect of NaAsO2 on the binding levels of methyl CpG binding protein 2(MeCP2),DNA methyltransferase 1 (DNMT1) and histone deacetylase 1(HDAC1) to the hypermethylation promoter region of MGMT gene in HaCaT cells,in order to provide a basis to deepen the interpretation of the role of arsenic poisoning mechanism.Methods HaCaT cells were treated repeatedly and interval with different concentrations of NaAsO2(3.13,6.25,12.50,25.00 μnol/L,respectively) for 72 h.Untreated HaCaT was used as blank control group and human epidermal squamous carcinoma cell line(A431 cells) as positive control group.The binding levels to the two transcription regulatory regions(ChIP1,ChIP2) and to the coding region(ChIP3) of MGMT 8ene were detected by chromatin immuno-precipitation combined with quantitative PCR.Results The differences of binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each group were significant (F=7.387,84.634,78.442 and 19.263,69.649,26.546,all P ＜ 0.05).The binding levels of MeCP2,DNMT1 and HDAC1 to ChIP1 and ChIP2 in each NaAsO2 exposed group[3.13 μmol/L NaAsO2 exposed group:(136.00 ±16.97)％,(145.00 ± 2.83)％,(88.50 ± 19.09)％ and (106.50 ± 37.48)％,(112.34 ± 8.73)％,(59.71 ± 8.49)％;6.25 μmol/L NaAsO2 exposed group:(130.00 ± 42.43)％,(154.50 ± 4.95)％,(101.00 ± 1.27)％ and (88.50 ±3.54)％,(134.32 ± 2.82)％,(102.75 ± 19.91)％ ; 12.50 μmol/L NaAsO2 exposed group:(141.50 ± 23.33)％,(161.50 ± 7.78)％,(125.00 ± 11.31)％ and (119.50 ± 24.75)％,(171.59 ± 3.54)％,(167.61 ± 10.61)％; 25.00μmol/L NaAsO2 exposed group:(134.50 ± 43.13)％,(472.50+ 50.20)％,(383.50 ± 30.41)％ and (180.09 ±12.73)％,(348.50 ± 27.58)％,(158.45 ± 12.02)％] were higher than that in the blank control group[(51.50 ±9.19)％,(82.00 ± 12.73)％,(25.03 ± 2.91)％ and (37.02 ± 4.24)％,(91.56 ± 26.16)％,(19.09 ± 2.90)％,all P ＜ 0.05].The differences of binding levels of MeCP2 to ChIP3 in each group were not significant(F =1.670,P ＞0.05),but the differences of binding levels of DNMT1 and HDAC1 to ChIP3 were significant (F =4.404,9.863,all P ＜ 0.05),and only the binding levels in the 25.00 μmol/L NaAsO2 exposed group [(615.85 ± 29.63)％,(306.09 ± 59.40)％] were higher than that in the blank control group[(99.70 ± 12.02)％,(92.45 ± 48.79)％,all P ＜ 0.05].Conclusions MeCP2 can bind to the methylated MGMT gene transcriptional regulatory regions which are induced by arsenic and leads to histone deacetylation by the recruitment of DNMT1 and HDAC1 and,meanwhile,DNMT1 can bind to the coding region of MGMT gene to recruit HDAC1 in a methyl DNA binding protein(MBD) independence manner and media MGMT gene silencing through the chromatin remodeling way,which might be the early molecular events of arsenic poisoning.

Child ; China ; Humans ; Paramyxoviridae Infections ; physiopathology ; Respiratory Tract Infections ; physiopathology ; virology

Adult ; Aortic Rupture ; pathology ; Death, Sudden, Cardiac ; etiology ; Heart Ventricles ; pathology ; Humans ; Male

Objective To investigate the mRNA,protein expression and tyrosine phosphorylation of insulin receptor substrate.1(IRS-1)in endometrial carcinoma.Methods Sixty-three endometrial carcinoma(EC)patients,21 endometrial atypical hyperplasia(AHE)patients and 22 normal control(NE) entered this smdy.Their clinical information were colleeted Fasting serum C-peptide concentration was measured.Expression of IRS-1 in endometrium was examined by RT-PCR and western blol Immunoprecipitation was used to measure the tyrosine phosphorylation of IRS-1.Results C-peptide 0.007].There were no significant differences in IRS-l mRNA and protein expression among the three grouDs.Tyrosine phosphorylation of IRS-1 in EC group[(62±36)%]was higher than that in AHE and NE groups[(53 4-34)%and(35 4-33)%;P=0.048,0.002].IRS-1 activation in AHE group was also higher than nornlal control(P=0.045).IRs-1 activation in endometrioid carcinoma[(69 4-33)%]was higher than that in other histological types[(34±31)%;t=2.300,P=0.025].IRs-l tyrosine phosphorylation was significantly higher in patients with advanced stage,high grade,deep myometrial invasion and pelvic lymph node metastasis.IRS-l activation in endometrium was positively correlated with fasting sertlm C-peptide concentration(r=0.491,P=0.001).Conclusions There is excessive activation of IRS-1 in endometrial carcinoma and atypical hyperplasia.Activation of IRS-I in endometrial carcinoma is related with poor clinical-pathologic fleatures and may be a prognostic predictor for this tumor.Over-activationof IRS-1 may be an intermediate event linking the hyperinsulinemia and endometrial carcinoma.