Aim To investigate the effect of curcumin on radiosensitivity of radioresistant nasopharyngeal carcinoma cell line CNE-2R and its mechanism.Methods The concentration of curcumin was screened by MTT assay.Dose-survival curves were obtained according to the colony forming test for L-Q matching and multitarget-single hitting matching,while SF2 and the correlation parameters of radiation biology were calculated.The changes of cell cycle in CNE-2R cells caused by curcumin were also tested by flow cytometry(FCM).The differential expression of genes related to cell cycle and DNA damage repair were detected by RT-qPCR.Results CNE-2R cells could not be inhibited by 10 μmol·L-1 curcumin.Dealt with 10 μmol·L-1 curcumin for 24 h,the value of α/β increased to 1 596 from 6.56;the value of SF2 decreased to 0.361 Gy from 1.93 Gy;the value of N decreased to 1.06 from 1.60;the value of D0 decreased to 2.12 from 3.27;the value of Dq decreased to 0.12 from 1.53.FCM showed that the cells in G2 phase had a significant increase and the cells in S phase had a significant decrease after dealt with 10 μmol·L-1 curcumin for 24 h.The expression of CDK4 was significantly up-regulated and GADD45g,BRCA1 were significantly down-regulated.Conclusion Curcumin radiosensitizes nasopharyngeal carcinoma cell line CNE-2R by changing cell cycle and affecting DNA damage repair through regulating the expression of CDK4,GADD45 g and BRCA1.

OBJECTIVETo investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer (PCa).

METHODSThe promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS-T carrier to verify it.

RESULTSThere were 5 polymorphisms. TAAA repeat times: 4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and ≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [OR=1.74, 95% CI=1.06-2.87 (for type 11TAAA); OR=5.63, 95% CI=1.85-17.19 (for type ≥12TAAA)]. In the 186 PCa patients, there was 62.4% allele of PCA3 gene with AG/CA mutation found in the promoter 18-19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer.

CONCLUSIONSShort tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Antigens, Neoplasm ; genetics ; metabolism ; Base Sequence ; Genes, Neoplasm ; physiology ; Genotype ; Humans ; Leukocytes, Mononuclear ; Male ; Microsatellite Repeats ; Mutation ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk

Amphetamine-type stimulants ( ATS ) , a group of new-type synthetic drugs mainly in psychological dependence, are abused more and more severely in recent years. MicroRNAs ( MiRNAs ) are an important class of endogenous non-coding small RNAs that mediate posttranscriptional negatively regulation of gene expression by targeting specific mRNA sequences to in-hibit the translation of mRNAs or degrade the expression of mR-NAs. ATS can induce the changes in the expression of miRNAs in addiction-related brain regions which directly involve in the regulation of ATS-induced addictive behaviors. Therefore, to study the regulatory role of miRNAs in ATS-induced addiction has important implications for dependent mechanisms of new-type drugs and the discovery of the new targets of drug actions.

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Objective To compare the effects of propofol,nidazolarm versus etomidate combined with sufentanil for anesthesia induction on intraocular pressure.Methods Forty-five ASA Ⅰ or Ⅱ patients,aged 20-40 yr,scheduled for surgery under general surgery,were randomly divided into 3 groups(n =15 each):propofol group(group P); midazolam group(group M)and etomidate group(group E).Anesthesia was induced with iv injection of propofol 2 mg/kg,midazolam 0.2 mg/kg,and etomidate 0.3 mg/kg in P,M and E groups respectively,and then with iv injection of sufentanil 0.2 μg/kg and cisatracurium 0.2 mg/kg in all the groups.The patients were then tracheal intubated.Intraocular pressure(IOP)and MAP were recorded at 1 m in before induction of anesthesia (T0),before intubation(T1),and at 0,1 and 2 min after intubation(T2-4).Results Compared with group P,the incidence of intraocujar hypotension was significantly decreased in group M(P ＜ 0.01).Compared with group E,the incidence of intraocular hypertension was significantly decreased in P and M groups(P ＜ 0.05),The correlation coefficient between MAP and IOP was 0.831,0.889 or 0.806 in group P,M or E respectively(P ＜0.05),and there was no significant difference in the correlation coefficient among the three groups(P ＞ 0.05).Conclusion Midazolam combined with sufentanil for anesthesia induction exerts less influence on lOP and the degree of MAP fluctuations is a major factor contributing to the change in IOP.

ObjectiveTo investigate the relationship of lymphotoxin β receptor (LTβR) and classical nuclear factor-κB (NF-κB) activation pathway in the pathogenesis and progress of cystitis and bladder cancer.MethodsThe LTβR and P65 mRNA expression were detected by Real-time quantitative PCR in 108 cases of fresh bladder tissue specimens (75 cases of bladder cancer,10 cases of inflammation and 23 normal bladder mucosa cases grouped by the tissue classification ),and protein expression were analyzed by immunohistochemistry assay in 118 cases of paraffin-embedded biopsy specimens (73 cases of bladder cancer,30 cases of cysitis and 15 normal bladder mucosa cases).The correlation analysis between the expressions of LTβR and P65 with clinical pathological data was then performed.Differences between LTβR and P65 mRNA and protein expression level were compared in different groups of bladder tissues using Kruskal-Wallis H test and the Chi-square test.Results( 1 )The mRNA expressions of LTβR and NF-κB/P65were higher in bladder cancer than those in normal group ( LTβR:29.4 ( 14.2 - 46.7 ) × 10 - 3/1.2 ( 0.3 -7.0) ×10-3,Z=-5.508; P65:9.7 (2.7 -21.1) ×10-3/1.0(0.8 ～1.8) ×10-3,Z=-5.030,P＜0.05 ).There were significantly differences between bladder cancer with different histological grades ( LTβR:18.2(2.1-31.3) × 10-3/ 28.4(16.6-36.2) × 10-3/47.9(34.3 -70.5) ×10-3,x2K-W=20.378;P65:4.9(1.3 - 12.0) × 10-3/7.4(3.0-21.9) × 10-3/17.0(10.0 ～28.3)× 10-3 ,x2K-W2 =15.219,P all ＜0.05) and lymph node metastasis (LTβR:27.2(9.7-40.1) ×10-3/39.4(26.7 -52.6) ×10-3,Z=-2.552; P65:7.4(2.3-15.6) ×10-3/13.4(6.7-23.3) ×10-3,Z=-2.026,P＜0.05).(2)The positive rates of LTβR and phosphorylated P65 ( p-P65 ) protein in cancer were higber than those of normal group (LTβR:69.8％/13.3％,x2 =16.600 ; p-P65:56.2％/6.7％,x2 =12.220,P ＜ 0.05 ).Upregulation of LTβR and p-P65 were associated with the histological grade (LTβR:56.3％/70.0％/90.4％,x2 =7.055; p-P65:40.6％ /60.0％/76.2％,x2 =6.679,P ＜0.05) and with lymph node metastasis (LTβR:58.3％/92.0％,x2 =8.849; p-P65:52.1％/64.0％,x2 =5.088,P ＜0.05).(3)There was a positive correlation between LTβR and P65 expression ( mRNA:r =0.654,P ＜ 0.05,protein:r =0.399,P ＜ 0.05 )in the bladder cancer and cystitis (r =0.521,P＜0.05).ConclusionsThe activation of LTβR and P65 was associated with progression and metastasis of bladder cancer.The activation of classical NF-κB pathway by LTβR may be achieved by P65.

Objective To establish a methamphetamine-dependent model in zebrabfish.Methods On the basis of conditioned place preference (CPP) in drug-dependent experiment,place preference box for zebrafishes was designed.According to the natural characteristics of zebrafishes,their preference side and non-preferred side were determined.After intraperitoneal injection of methamphetamine,zebrafishes were placed in non-preferred side(drug box).After intraperitoneal injection of normal saline,zebrafishes were placed in preference side (non-drug box).CPP training was five days and then the methamphetamine-induced place preference in zebrafish were observed.Results After five days training,the staying time of zebrafishes of control group in drug box was not significantly lengthened to compared with before training(.( 287.5 ± 80.18 ) s,(276.3 ± 85.04) s),P＞ 0.05 ).The staying time of zebrafishes of model group in drug box was markedly extended after training.In comparison with before training or control group,the significant differences were observed ( (465.5 ± 113.49 ) s,( 247.9 ±95.62)s,(276.3 ±85.04)s,P＜0.01).Conclusion Methamphetamine can induce conditioned place preference in zebrafishes.The CPP model in zebrafish established in this study can be used as a new animal model in drug dependence.

Objective To study changes of AMPA receptors expression in nucleus accumbens and hypothalamus of methamphetamine dependent rats,and the therapeutical effect of rhynchophylline.Methods SPF male rata were randomly divided into normal control group,model group of methamphetamine,low dose of rhynchophylline group and high dose of rhynchophylline group(n=8 in each group).Experiment of conditioned place preference(CPP)was used to build the model of methamphetamine dependent rata.Western blotting was used to examine the changes of GluR2/3 subunits expression.The time of staying in drug-paired compartment of rats was used independent-samples t test to gather statistics,and the photodensity of proteinum strap was used One-Way ANOVA to gather statistics.Results Compare with rats in normal control group(the time of staying in drug-paired compartment of rats was(383.00±38.20)s),the rats produced CPP after treated with methamphetamine(the time of staying in drug-paired compartment of rats was(536.20±57.49)s),and low(30mg/kg) and high (60 ms/kg)dose of rhynchophylline(the time of staying in drug-paired compartment of rats were(299.80±15.96)s and(189.40±59.02)s)both could eliminate CPP effect.Compare with rats in normal control group (the ratio of value of average gray scale were(0.54±0.04)INT·mm~2 and (0.70±0.04)INT·mm~2),GluR2/3 subunits expression in nucleus aecumbens increased significantly in model group(the ratio of value of average gray seale was(0.89±0.03)INT·mm~2)and low dose of rhynchophylline group(the ratio of value of average gray seale was (0.93±0.03)INT·mm~2,P<0.01),which decreased significantly in hypothalamus(the ratio of value of average gray scale were (0.53±0.03)INT·mm~2 and (0.52±0.02)INT·mm~2,P<0.01).But GluR2/3 subunits expression in nucleus accumbens and hypothalamus of rats in high dose of rhynchophylline group(the ratio of value of average gray scale were (0.57±0.06)INT·mm~2 and (0.65±0.01)INT·mm~2) just liked the expression of normal control group(P>0.05).Conclusion GluR2/3 subunits expression of methamphetamine-induced CPP rats increased in nucleus accumbens but decreased in hypothalamus.High dose of rhynchophylline can reverse such changes and rebound the expression to normal level.

Objective To investigate the changes of monoamine neurotransmitters of the brain in physical dependence induced by morphine in rats.Methods A physical dependent rat model was established with morphine in a gradually in creasing doses and the withdrawal syndrome was scored after naloxone precipitation.The conditioned place preference(CPP)in rats induced by morphine was used to investigate psychic dependence in rats.Contents of norepinephrine(NE0,dopamine(DA)and serotonin(5-HT)in hypothalamus of rats were assayed with a fluorescent method. Results (1)In naloxone-precipitated withdrawal test of morphine-dependent model rats,after morphine had been withdrawn,morphine-abstinent rats presented marked withdrawal symptoms and signs,their withdrawal scores were significantly increased,and the levels of NE and 5-HT in the rat brain were obviously enhanced,but the content of DA was reduced.(2)In CPP test,morphine caused a marked place preference in rats and the levels of DA and 5-HT in the rat brain were obviously enhanced,but the content of NE was reduced.Conclusiion Morphine dependence development and withdrawal are closely connected with monoamine neurotransmitters in CNS.In the physical dependent model induced by morphine in rats,the rising of NE and 5-HT in the rat brain were significant,but in psychic dependent model induced by morphine in rats,the levels of DA in the rat brain were enhanced p-redominately.