Objective To discuss some characteristics of aggressive behavior of junior middle school students addicted to violence online game.Methods From Aug.to Dec.at 2009,6000 volunteers have been investigated with the self-made violence online game survey scale and the Chen's were screen out by CIAS,and in which 71 violence game addicts,and 66 entertainment game addicts.They were divided into two groups matched as age and sex(male 55 and female 11 in every group),average age was 14.36±0.81 years old.66 no-play game students served as control group.The mean value and standard deviation of aggressive behaviors in each group were compared with one way ANOVA with SPSS12.0 version.Results There were significant different inter groups in the behavioral scores expressed in physiological attack((21.59±4.36),(18.10±6.10,(14.92±5.61)),spoken language attack((21.04±3.98),(18.36±6.26),(15.66±5.23)),angry mood((21.37±4.43),(18.72±5.90),(16.56±5.70)),hostility cognition((21.69±5.77),(20.06±7.74),(16.39±6.85))and the total score((85.56±13.71),(75.28±22.36),(63.54±20.32))(F1-4=25.07,17.35,3.25,10.40,21.81);Moat severity attack behaviors were observed in the violence online game addictive group,but less done in other two groups(t1-4=3.48,2.68,2.65,10.27,P<0.01;t 1-5=6.66,5.37,4.48,5.30,22.01,P<0.01);and secondary was found in Entertainment online game addicts(t1-5:3.18,2.69,2.16,3.66,11.74,P<0.01).Conclusion Higher attack behavior scores are found in the violence online game in junior middle students,and entertainment online game also induce attack behavior,but is not so notable.

AIM: To discuss the relationship between prostate specific membrane antigen(PSMA) and prostate cancer and to seek a target for diagnosis and therapy of prostate cancer.METHODS: A pair of primers was designed according to the published PSMA mRNA sequence.Total RNA was extracted from prostate cancer tissues and was reversely transcribed into cDNA,which was used as a template for PCR to amplify the PSMA gene.The recombinant was sequenced and the result was analyzed by BLAST.The PSMA5 gene specific primers were designed to identify its expression in different cells and prostate tissues.RESULTS: A new alternatively spliced variant of PSMA named PSMA5 was discovered when sequencing the recombinant.PSMA5 showed well pathological tissue-specificity,and its expression rate in prostate cancer,benign prostatic hyperplasia of prostate,and normal prostate tissue were 92.6%,78.8% and 10.0%,respectively.It expressed specifically in Pca cell line LNCaP,not in cell lines of PC3,bladder carcinoma,renal carcinoma,or hepatoma.CONCLUSION: A new alternative spliced variant of PSMA named PSMA5 was discovered,which was well correlated with prostate cancer and benign prostatic hyperplasia.This finding may give a new clue to the evolution of prostate cancer and may provide a target for the diagnosis and therapy of prostate cancer.

AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.

AIM:To study the effects and mechanisms of ethanol on chloride channels in poorly differentiated nasopharyngeal carcinoma CNE-2Z cells.METHODS:The effect of ethanol on the cell growth was analyzed by MTT as-say.The technique of whole-cell patch-clamp was used to detect the chloride current .The characteristics of the chloride current were analyzed by using the chloride channel blockers .The siRNA technique was used to analyze the molecular basis of the ethanol-sensitive chloride channels .RESULTS: Under isotonic conditions , the background current was weak and stable.Ethanol at concentrations of 0.17~170 mmol/L activated a chloride current in a concentration-dependent manner (an inverted U-shape), with a maximum effect at the concentration of 17 mmol/L.The currents showed obviously outward rectification and were susceptible to extracellular hypertonicity and the chloride channel blocker , 5-nitro-2-(3-phenylpropyl-amino) benzoic acid ( NPPB) .ClC-3 siRNA obviously decreased the currents activated by ethanol .CONCLUSION:Ex-tracellular ethanol induces chloride currents through activating the ClC-3 chloride channels .

OBJECTIVETo construct four expression vectors carrying enhanced green fluorescent protein (EGFP) gene under the control of different HBV promoters, and to detect and analyze their expressions in hepatoma cell lines.

METHODSFour HBV promoters were amplified using PCR, and they were inserted into the T-vector and identified using restriction enzymes and sequencing, then cloned into the expression vector pEGFP-1. The four recombinant plasmids were transfected into human hepatoma cell line HepG2 by lipofectamine2000, and the positive cell clones were detected using fluorescence microscopy.

RESULTSAll target fragments were separately obtained and successfully cloned into the expression vector. The expressions of EGFP under the control of the four promoters were detected. The expressions of EGFP controlled by different promoters had some differences.

CONCLUSIONSReporter gene EGFP under the control of four HBV promoters can be specifically expressed in hepatoma cell line HepG2, and different promoters give different results; this may provide another option in gene therapy of liver diseases.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Eukaryotic Cells ; metabolism ; Genetic Therapy ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HeLa Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; Promoter Regions, Genetic ; genetics ; Transfection

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

Objective To discuss family environment and parent's education pattern of violent online games addictive disorder in junior middle school students. Methods From August to December at 2008,6000 students were randomly drown from 8 junior middle schools in Dalian. All subjects were investigated with the self-made internet game survey form and the Chen internet addiction scale (CLAS). 149(2.5% ) internet game addicts were screen out from 6000 students, and then were divided into the violence and entertainment game addicted group (66 students in per group, and the male 55, female 11). No-game group 66 students were matched according tothe sex and the age. All subjects were evaluated with Egma Minnen av Bardndosnauppforstran ( EMBU) and Moss's Family Environment Scale of Chinese version. Results Lower mother's understand and warm were observed in violence internet online game addictive student group than other two groups( (46. 37 ± 13. 93,49. 80 ± 7.13,53. 16 ± 14. 89) , F = 4.88, P<0.01) ,and the mother understand and warm were remarkably lower in the violence online game addictive group than the non-game addicted group(P<0.01) ,and mothers rejection and the denial are obviously higher in entertainment online game addictive group than the non-game group(P = 0. 038). In family environment survey results suggested there were significant different during the 3 groups (F1-3 = 13. 99, 12.24,10.56; P<0.05)in emotion expression(4. 74 ± 1. 32,6. 06 ± 1. 97,5. 80 ± 1. 11) ,and conflict (5. 57 ± 1.85,5.28 ± 1.41,3.63 ± 1.50) .independence (3.96 ± 1. 87,4.56 ± 1.84,4.63 ± 1. 85). In the violence online game addictive group, emotion expression was lower than other two groups (P<0. 01) ,the conflict score was higher than other two groups (P<0.01). The controlling was lower than other two groups ( all P<0. 01) ; family function and total score are lower in online game addictive groups than other groups (P<0. 01, P<0. 05). Conclusion -Lower emotion expression,higher conflict,lacks of mother's love and warm,and lower controlling are found in violence online game addictive junior middle school students.

Objective To discuss the personality characteristic and personal social attributes of junior middle school students of online violent games addictive disorder (VOGAD).Methods From August to December at 2008,6000 students were randomly drown from 8 junior middle schools in Dalian ,then they were investigated with the self-made internet game survey form and the Chen internet addiction scale (CLAS).149( 2.5% ) internet game addicts were screen out from 6000 students, and were divided into the violence and entertainment game addicted group ( 66 students in per group,and the male is 55, female is 11 ).Average age was ( 14.36 ± 0.81 ) years old.No-play game group 66 students were mach according to the sex and the age.All objects were evaluated with Eysenck personality questionnaire (EPQ) and Minnesota Multiphasic Personality Inventory Abbreviation Form (MMPI-AF).Results There were significant different during the 3 groups in psychoticism (50.60 ± 5.10;50.40 ± 6.30; 46.81 ± 4.62 ), neuroticism ( 51.74 ± 5.70; 49.03 ± 6.85; 49.25 ± 6.09 ) , manifest anxiety (55.29 ±9.58;45.11 ±8.31; 43.24 ±8.32) ,dependence (58.90 ±9.09; 55.97 ±8.63; 54.05 ± 11.39) ,and social responsibility ( 37.47 ± 12.41; 43.25 ± 11.24 ;46.34 ± 14.37 ) (F1-5 = 10.32,3.83,36.17,4.11,8.23; P 1-5 ＜ 0.05 ).Except of the social responsibility (P＞ 0.05 ) ,above results all suggested higher scores in the VOGAD group than other two groups (P1 ＜ 0.05 ).Conclusion The results suggest higher scores in the psychoticism, neuroticism, dependency, manifest anxiety in VOGAD group, and lower Re in online violence game addict students.

Objective To determine the protective role of keratinocyte growth factor 2 (KGF-2) on dextran sodium sulfate (DSS)-induced murine colitis and investigate the mechanism of its effect on intestinal mucosal barrier.Methods A total of 36 male C57BL/c mice were divided into 4 groups:normal control group,DSS model group,model + KGF-2 (5 mg/kg) group and model + KGF-2 (10 mg/kg)group.Groups except the control were added 3.5 ％ DSS in drinking water.Disease activity index (DAD,weight change,colon length loss and histological score were detected to evaluate the protective effect of KGF-2 on colitis.Serum fluorescein isothiocyanate-dextran (FITC-D) permeability were assayed by multiscan spectrum.In order to explore the protective role of KGF-2 on murine intestinal mucosal barrier,ZO-1 and occludin protein concentration were detected by immunohistochemical staining and Western blot.Meanwhile,cytokines including TNF-α,IL-6,IL-10,TGF-1β,IFN-γ and IL-1β in colonictissue were detected by ELISA.Results Compared with DSS-induced colitis model group,10 mg/kg KGF-2 significantly reduced DAI (P =0.021 1),weight loss (P =0.017 6),colon length loss (4.956 ±0.2583 vs.6.289±0.215 7,P =0.001 1),histologicalscore (12.17±1.222 vs.7.000±0.6325,P =0.001 1),and FITC-D permeability (168.5 ± 27.01 vs.14.62 ± 1.812,P =0.004 7) and reversed the downregulation of tight junction (TJ) proteins (ZO-1:0.158 6 ± 0.010 51 vs.0.387 9 ± 0.028 64,P＜0.000 1;occluding:0.300 5 ± 0.026 56 vs.0.445 0 ± 0.056 62,P =0.043 4).Significant decrease of TNF-α (68.93 ± 3.379 vs.40.41 ± 1.576,P＜0.000 1) and increase of IL-6 (3 755 ± 309.8 vs.5 640 ± 418.0,P =0.004 7) and IL-10 (304.0 ± 21.47 vs.521.2 ± 49.40,P =0.002 4) levels were noted in the 10 mg/kg KGF-2 treated mice.The effect of the KGF-2 was dose-dependent.Conclusions KGF-2 could ameliorate DSS-induced colitis and it may be associated with the decrease of the damage of mucosal barrier structure and funcntion by preventing ZO-1 and occludin from downregulating.The protective effect of KGF-2 on intestinal barrier function may also be exerted by inhibition of TNF-α and stimulation of IL-6 and IL-10 secretion.

It is a common understanding that turbidity and precipitation of traditional Chinese medicine are easy to occur in the process of decocting. At present, our research group found that the cause of "multi-phase of traditional Chinese medicine decoction" mainly came from the interaction between the effective components of traditional Chinese medicine, especially the interaction of acid and base components. For example, the Liquorice and Rhizoma chinensis was a supramolecular system formed by a large number of active components in the decoction (>30%), and could stably exist in the decoction system. In this study, the supramolecular part was extracted, and the morphology of the supramolecular part was characterized by scanning electron microscopy and dynamic light scattering. It was observed that the supramolecular particles were uniform in size and regular in shape. The main components of supramolecular sites were identified by liquid mass spectrometry (LC-MSⁿ). The results of UV and IR spectra showed that the chemical components of Liquorice and Rhizoma chinensis in the co-decocting process collided with each other, and weak bonds were formed between the functional groups of the molecules, which then induced the aggregation to form supramolecules. Thereafter, Through the diarrhea model of mice, sensory evaluation and antibacterial activity evaluation found that Liquorice and Rhizoma chinensis decocted together enhanced the antibacterial activity of Rhizoma, and compatibility "reconcile" Rhizoma "big bitter cold" property compared with single decoction group and interval administration group. All animal experiments were approved by the Animal Ethics Committee of Beijing University of Chinese Medicine, and the relevant regulations of Beijing University of Chinese Medicine on experimental animals were strictly followed. In this study, supramolecular chemistry method was used to preliminarily discuss the scientific connotation of "increasing efficiency and decreasing toxicity" of Liquorice and Rhizoma chinensis combined decoction from three perspectives of "property, efficacy and taste", and provide new ideas for the basic research of "reconcile" compatibility of Liquorice.