[Cloning and expression analysis of tocopherol cyclase gene in Carthamus tinctorius].

The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named CtTC . Bioinformatics analysis showed theopen reading frame （ORF）of CtTC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62.9 kDa and theoretically isoelectric point was 5.01.Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide. The subcellular localization showed that the CtTC protein was located in the chloroplast. The expression of CtTC gene in safflower seeds at different development stages was determined by quantitative real-time PCR, it was found that the highest expression level of CtTC gene was detected in 50 DAF.Quantitative RT-PCR analysis suggested that expression of CtTC is induced and strengthened by drought stresses. This research provided a candidate gene for metabolic engineering of vitamin E and resisting stress.

[Serum levels of 25-hydroxyvitamin D in children with attention deficit hyperactivity disorder].

OBJECTIVE: To examine serum 25-hydroxyvitamin D levels in children with attention deficit hyperactivity disorder (ADHD) and to explore the relationship between vitamin D level and ADHD. METHODS: Ninety-seven children with ADHD who were diagnosed according to DSM-V were selected as the ADHD group, including 46 cases of ADHD-I, 10 cases of ADHD-HI, and 41 cases of ADHD-C. Ninety-seven healthy children served as the control group. Serum levels of 25-hydroxyvitamin D were measured using electrochemiluminescence immunoassay. RESULTS: Mean serum levels of 25-hydroxyvitamin D in the ADHD group (17±7 ng/mL) were significantly lower than in the control group (23±8 ng/mL; P<0.01). The serum levels of 25-hydroxyvitamin D in the three subtypes groups of ADHD (ADHD-I, ADHD-HI, and ADHD-C) were all lower than in the control group (P<0.05). The rates of vitamin D insufficiency, deficiency or normal in the ADHD group were different from the control group (P<0.01). The distributions of vitamin D levels in the three subtypes groups of ADHD were all different from the control group (P<0.05). CONCLUSIONS: Serum levels of 25-hydroxyvitamin D in children with ADHD are lower than in healthy children, suggesting vitamin D level might be related to ADHD.

Upregulation of SCC-S2 in immune cells and tumor tissues of papillary thyroid carcinoma.

Studies have shown that SCC-S2 can be detected in cancer cells, but its relation with thyroid cancer remains uncertain. In the current study, we investigated SCC-S2 expression in thyroid cancer from the immune cell perspective and tumor tissue perspective. Levels of SCC-S2 in CD4+ T cells, CD8+ T cells, monocytes, natural killer (NK) T cells, tumor tissues, and adjacent noncancerous thyroid tissues were tested by real-time reverse transcription PCR and Western blot. Results revealed that mRNA and protein levels of SCC-S2 were significantly increased in peripheral CD4+ (mRNA, 1.90-fold; protein, 1.55-fold) and CD8+ T cells (mRNA, 2.37-fold; protein, 1.72-fold) but not monocytes and NKT cells in patients than in healthy donors. Further elevated mRNA level but not protein expression was observed in tumor-infiltrating CD4+ T cells, whereas both mRNA level and protein expression were further increased in tumor-infiltrating CD8+ T cells. Also, mRNA and protein levels of SCC-S2 in thyroid tissues were significantly elevated than those in adjacent noncancerous thyroid tissues. Moreover, patients with cervical lymph node metastasis presented clearly higher mRNA and protein expression of SCC-S2 compared to those without cervical lymph node metastasis (p < 0.05). These results suggest that SCC-S2 may play roles in affecting both immune cells and tumor cells in the thyroid and may indicate a novel pathway for understanding the pathogenesis of the disease.

[Establishment of the Chang liver cell line stably overexpressing human UCP2 gene and its effect on mitochondrial membrane potential and reactive oxygen species].

To establish the Chang liver cell line stably overexpressing human uncoupling protein 2 (UCP2) and observe the effect of UCP2 on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The Chang liver cell line was transfected with recombinant plasmid containing full-length human UCP2 cDNA (pcDNA3.1-hUCP2) or pcDNA3.1 empty vector. The stable cell line was established by antibiotic screening with Zeocin. UCP2 expression was detected by Western blotting and immunocytochemistry. The UCP2 overexpressing cells were pretreated with genipin at various doses (25, 50 and 100 munol/L). MMP and intracellular ROS were detected by fluorescence spectrophotometry. The total normalized protein content in UCP2 overexpressing cells was 1.6-fold higher than that in unmanipulated normal cells. The fluorescence intensities of Rhodamine123 and DCFH-DA in UCP2 overexpressing Chang liver cells (11.11+/-2.76 and 4.97+/-0.62, respectively) were significantly lower than those in unmanipulated normal cells (15.56+/-2.55, P less than 0.01 and 6.14+/-1.25, P less than 0.05, respectively) and in cells transfected with empty vector (16.11+/-2.93, P less than 0.01 and 6.23+/-1.13, P less than 0.05, respectively). Treatment of UCP2 overexpressing cells with 25, 50 and 100 munol/L genipin caused a dose-dependent increase in fluorescence intensities of Rhodamine123 (14.89+/-2.89, 17.89+/-2.93 and 24.00+/-2.55, respectively, all P less than 0.01) and DCFH-DA (9.16+/-0.78, 10.84+/-1.09 and 11.83+/-1.25, respectively, all P less than 0.01). The Chang liver cell line stably overexpressing UCP2 was established successfully. Using this cell system, UCP2 was found to play a role in mitochondrial function by regulating MMP and ROS.

Association analyses of MAOA in Chinese Han subjects with attention-deficit/hyperactivity disorder: family-based association test, case-control study, and quantitative traits of impulsivity.

Monoamine oxidase A (MAOA) plays a critical role in the metabolism of monoamine neurotransmitters including serotonin (5-HT), norepinephrine (NE), and dopamine (DA). Genetic studies have found an association between MAOA and attention-deficit/hyperactivity disorder (ADHD), especially impulsivity. However, there has been inconsistency among studies which may be due to the complexity and heterogeneity of ADHD, including its sexual dimorphism and the presence of several subtypes. We conducted transmission disequilibrium tests (TDTs) in 1,253 trios and found no association between five single nucleotide polymorphisms (SNPs) of MAOA with ADHD in general or in the predominantly inattentive (ADHD-I) or combined types (ADHD-C), but with the predominantly hyperactive/impulsivity type (ADHD-HI). The association with MAOA was restricted to males, especially males with ADHD-HI. Logistic regression analyses of data from 1,824 cases and 957 controls did not indicate any association. We used analysis of covariance to analyze the association between MAOA genotype with the "inhibit" factor of the Behavior Rating Inventory of Executive Function (BRIEF) in 640 probands and performance on the Stroop test in 810 probands. Probands homozygous for risk alleles found in the TDT test had higher "inhibit" scores on the BRIEF scale which represents more severe impulsivity; this results also was restricted to males. No association was found with Stroop test performance. In conclusion, our results provide some evidence that MAOA may be associated with the ADHD-HI subtype and support the association between MAOA and impulsivity, which may be a potential endophenotype of ADHD. However, the results were strongly influenced by gender.

Preparation, Characterization and Pharmacokinetic Study of N-Terminal PEGylated D-Form Antimicrobial Peptide OM19r-8.

Recently, new cationic antibacterial peptide OM19R has been designed with low minimum inhibitory concentration (MIC) values against some gram-negative bacteria, such as Escherichia coli, Salmonella, and Shigella. However, this hybrid peptide, like most antibacterial peptides, has low enzyme stability and short half-life, which, in turn, increases the drug's cost. In this study, an antibacterial peptide (OM19r-8) was obtained containing some D-Arg amino acids. The new preparations were carried out through the replacement of l-Arginine by d-Arginine and the addition of PEG chains. Firstly, eight OM19r series of antibacterial peptides were obtained by designing D-Arg. Then, a polyethylene glycol-modified product mPEG5-butyrALD-OM19r-8 (mPEG5-OM19r-8) was isolated and purified by reverse-phase high-performance liquid chromatography (RT-HPLC). The enzyme stability test showed that the resistance of antibacterial peptide OM19r-8 to protease degradation increased by 4-32-fold. Moreover, the Time-kill studies showed that the germicidal kinetics curves of mPEG5-OM19r-8 and OM19r-8 to Escherichia coli had a similar trend, thus suggesting that PEG modification has an acceptable effect on the activity of the original peptide. Furthermore, the elimination of half-life (28.09 ± 2.81min) of mPEG5-OM19r-8, and the area under the drug concentration-time curve (2686.48 ± 651.36min∗ug/ml) was significantly prolonged. The current study demonstrates an example that optimizes the AMP by utilizing L-to-D amino acid replacement and including PEG chains. These results provide useful data for the clinical application of the mPEG5-OM19r-8.

Attenuated and delayed niacin skin flushing in schizophrenia and affective disorders: A potential clinical auxiliary diagnostic marker.

AIM: Schizophrenia and affective disorders all show high heterogeneity in clinical manifestations. A lack of objective biomarkers has long been a challenge in the clinical diagnosis of these diseases. In this study, we aimed to investigate the performance of niacin skin flushing in schizophrenia and affective disorders and determine its clinical potential as an auxiliary diagnostic marker. METHODS: In this case-control study, niacin skin-flushing tests were conducted in 613 patients (including 307 schizophrenia patients, 179 bipolar disorder patients, and 127 unipolar depression patients) and 148 healthy controls (HCs) with a modified method. Differences in niacin skin-flushing responses were compared with adjustment for gender, BMI, age, nicotine dependence, alcohol consumption and educational status. A diagnostic model was established based on a bivariate cut-off. RESULTS: Schizophrenia and affective disorders showed similar performance of niacin bluntness, characterized by attenuated flushing extent and reduced flushing rate. An innovative bivariate cut-off was established according to these two features, by which we could identify -patients with either schizophrenia or affective disorders from HCs with a sensitivity of 55.28%, a specificity of 83.56% and a positive predictive value of 93.66%. CONCLUSIONS: The niacin-induced skin flushing was prevalently blunted in patients with schizophrenia or affective disorders, indicating a promising potential as an auxiliary diagnostic marker in risk prediction and clinical management of these disorders. Additionally, the niacin-blunted subgroup implies a common biological basis in the investigated disorders, which provokes new thoughts in elucidating the pathological mechanisms.

Gene-gene interaction between COMT and MAOA potentially predicts the intelligence of attention-deficit hyperactivity disorder boys in China.

The catechol-O-methyltransferase (COMT) gene contains a functional polymorphism (Val158Met) affecting the activity of the enzyme, and the monoamine oxidase A (MAOA) gene contains a VNTR polymorphism (MAOA-uVNTR) that affects the transcription of the gene. COMT and MAOA each contribute to the enzymatic degradation of dopamine and noradrenaline. Prefrontal cortical (PFC) function, which plays an important role in individual cognitive abilities, including intelligence, is modulated by dopamine. Since our previous association studies between attention deficit hyperactivity disorder (ADHD) and these two functional polymorphisms consistently showed the low activity alleles were preferentially transmitted to inattentive ADHD boys, the goal of the present study was to test the hypothesis that the interaction between COMT Val158Met and MAOA-uVNTR may affect the intelligence in a clinical sample of Chinese male ADHD subjects (n = 264). We found that the COMT x MAOA interaction significantly predicted full scale (FSIQ) and performance (PIQ) IQ scores (P = 0.039, 0.011); the MAOA-uVNTR significantly predicted FSIQ, PIQ and verbal IQ (VIQ) (P = 0.009, 0.019, 0.038); COMT Val158Met independently had no effect on any of the IQ scores. Only the COMT x MAOA interaction for PIQ remained significant after a Bonferroni correction. Among all combined genotypes, the valval-3R genotype predicted higher intelligence, (average 106.7 +/- 1.6, 95% C.I. 103.7-109.8 for FSIQ), and the valval-4R predicted lower intelligence (average 98.0 +/- 2.3, 95% C.I. 93.5-102.6 for FSIQ). These results suggest that there is an inverted U-shaped relationship between intelligence and dopaminergic activity in our sample. Our finding that gene-gene interaction between COMT and MAOA predicts the intelligence of ADHD boys in China is intriguing but requires replication in other samples.

[Serum omega-3 polyunsaturated fatty acid and insulin resistance in type 2 diabetes mellitus and non-alcoholic fatty liver disease].

OBJECTIVE: To investigate the relationship between serum omega-3 polyunsaturated fatty acid (omega-3PUFA) and insulin resistance (IR) in patients with type 2 diabetes mellitus and non-alcoholic fatty liver disease (NAFLD). METHODS: This trial involved 51 patients of type 2 diabetes mellitus with NAFLD (G4 group), 50 patients of type 2 diabetes alone (G3 group), 45 patients of NAFLD alone (G2 group) and 42 healthy control subjects (G1 group). Serum omega-3PUFA profile was analyzed with capillary gas chromatography. Insulin resistance was assessed by homeostasis model assessment (HOMA-IR). ALT, AST, gamma-glutamyltransferase (GGT) and serum lipids were measured. RESULTS: The levels of HOMA-IR were higher in G4 group than those in G3, G2 and G1 group (4.90 + or - 2.54 vs 2.38 + or - 1.23, 2.20 + or - 1.15, 1.13 + or - 0.42; P < 0.05). The level of ALT, AST, GGT, TC, TG, LDL-C were higher in G4 group than those in G3, G2 and G1 group (P < 0.05). The level of omega-3PUFA was significantly lower in G4 group than those in G3, G2 and G1 group (5.68 + or - 2.02 vs 7.17 + or - 2.38, 6.97 + or - 2.32, 10.08 + or - 2.76; P < 0.05). omega-3PUFA concentration was negatively correlated with HOMA-IR, TC, TG and LDL-C (r = -0.491, -0.376, -0.462, -0.408, P < 0.05). CONCLUSIONS: Serum omega-3PUFA is significantly decreased in patients with type 2 diabetes mellitus and NAFLD. Serum omega-3PUFA is negatively correlated with insulin resistance. omega-3PUFA plays a very important role in the development of diabetes mellitus and NAFLD.

Identification and characterization of human ARIP2 and its relation to breast cancer.

Activin, a member of the TGF-beta superfamily, inhibits the proliferation of breast cancer cells. Activin interacts with its type I and type II receptors to induce phosphorylation of intracellular signaling molecules known as Smads. Previous studies showed that mouse ARIP2 can reduce activin signaling by interacting with activin type II receptors (ActRIIs); however, the activity of ARIP2 in breast cancer is still unclear. In this study, we used RT-PCR to obtain a human homologue of mouse ARIP2, human activin receptor-interacting protein 2 (hARIP2). Like murine ARIP2, hARIP2 has a PDZ domain in its NH2-terminal region and can interact specifically with ActRIIs. Overexpression of hARIP2 reduced activin-induced transcriptional activity and enhanced cell proliferation and colony formation in human breast adenocarcinoma MCF-7 cells and MDA-MB-231 cells. However, down-regulation of hARIP2 expression by RNAi enhanced activin-induced transcriptional activity and reduced cell proliferation and colony formation. Immunohistochemistry revealed that hARIP2 was expressed more frequently and much more intensely in malignant breast tissues such as simple carcinoma, invasive ductal carcinoma and mucinous adenocarcinoma than in benign hyperplasia or fibroadenoma cases. These results suggest that hARIP2 is a putative growth-promoting factor involved in breast tumorigenesis and tumor development.

Attention Deficit Hyperactivity Disorder comorbid oppositional defiant disorder and its predominately inattentive type: evidence for an association with COMT but not MAOA in a Chinese sample.

BACKGROUND: There are three childhood disruptive behavior disorders (DBDs), attention deficit hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), and conduct disorder (CD). The most common comorbid disorder in ADHD is ODD. DSM-IV describes three ADHD subtypes: predominantly inattentive type (ADHD-IA), predominantly hyperactive-impulsive type (ADHD-HI), and combined type (ADHD-C). Prior work suggests that specific candidate genes are associated with specific subtypes of ADHD in China. Our previous association studies between ADHD and functional polymorphisms of COMT and MAOA, consistently showed the low transcriptional activity alleles were preferentially transmitted to ADHD-IA boys. Thus, the goal of the present study is to test the hypothesis that COMT Val158Met and MAOA-uVNTR jointly contribute to the ODD phenotype among Chinese ADHD boys. METHODS: 171 Chinese boys between 6 and 17.5 years old (mean = 10.3, SD = 2.6) with complete COMT val158met and MAOA-uVNTR genotyping information were studied. We used logistic regression with genotypes as independent variables and the binary phenotype as the dependent variable. We used p < 0.05 as the level of nominal statistical significance. Bonferroni correction procedures were used to adjust for multiple comparisons. RESULTS: Our results highlight the potential etiologic role of COMT in the ADHD with comorbid ODD and its predominately inattentive type in male Chinese subjects. ADHD with comorbid ODD was associated with homozygosity of the high-activity Val allele, while the predominantly inattentive ADHD subtype was associated with the low-activity Met allele. We found no evidence of association between the MAOA-uVNTR variant and ADHD with comorbid ODD or the ADHD-IA subtype. CONCLUSION: Our study of attention deficit hyperactivity disorder comorbid oppositional defiant disorder and its predominately inattentive type highlights the potential etiologic role of COMT for ADHD children in China. But we failed to observe an interaction between COMT and MAOA, which suggests that epistasis between COMT and MAOA genes does not influence the phenotype of ADHD-IA with comorbid ODD in a clinical sample of Chinese male subjects. To confirm our findings further studies with a larger number of subjects and healthy controls are needed.

[Association study of intelligence of attention deficit hyperactivity disorder children in China].

OBJECTIVE: To investigate the association of 5-HT(2A) receptor gene (HTR2A)-1438A/G, Catechol-O-methyltransferase (COMT) gene Val158Met, Monoamine oxidase A ( MAOA ) gene 30 bp-VNTR(MAOA-uVNTR)polymorphisms, and the educational attainment level of the parents with the intelligence of attention deficit hyperactivity disorder (ADHD) in China. METHODS: A total of 485 DSM-IV ADHD children of Chinese Han descent were included, both complete IQ evaluation, HTR2A-1438A/G, COMT gene Val158Met, and MAOA-uVNTR genotyping results were obtained. The quantitative traits of psychometric IQ were calculated by using the Chinese Wechsler Intelligence Scale for Children (C-WISC). The multifactor linear regression analysis was used to test the associated factors on intelligence. RESULTS: Analyses revealed that ADHD children with low enzymatic activity (3R for males, 3R3R for females) of MAOA-uVNTR performed better on Full Scale IQ (FIQ) than did patients with high enzymatic activity (4R for males, 3R4R/4R4R for females) [(102.6+/-12.4) vs (100.3+/-11.7), P=0.078]. The patients with high-enzymatic activity (ValVal) of COMT gene Val158Met performed significantly better on FIQ than did patients with mid-low enzymatic activity (ValMet and MetMet)[(103.5+/-13.6) vs (100.5+/-11.5), P=0.036]. ADHD children with GG genotype of HTR2A-1438A/G performed significantly better on some aspects of C-WISC test (Full Scale IQ and Verbal Scale IQ) than did children with GA and AA genotypes [FIQ :(106.9+/-10.7) vs (100.7+/-12.3) vs (101.7+/-12.9), P=0.003; VIQ: (110.1+/-10.6) vs (103.5+/-12.1) vs (105.1+/-13.2), P=0.001]. The educational attainment level of the parents was associated with all the aspects of C-WISC test (Full Scale IQ, Verbal Scale IQ, and Performance Scale IQ). The multiple linear regression analysis showed that the genotype of HTR2A-1438A/G had significant correlation with FIQ, VIQ and PIQ; while the educational attainment level of the mother had significant correlation with FIQ and VIQ. CONCLUSION: The HTR2A-1438A/G polymorphism and the educational attainment level of the mother were associated with the intelligence of ADHD children in China.

[Effects of Genipin on the expression of uncoupling protein 1 in brown adipose and white adipose tissues in mice].

OBJECTIVE: To investigate the effects of genipin on promoting brown adipose tissue activation and white adipose tissue browning. METHODS: The male C57BL/6J mice were divided into three groups: normal control group, genipin group and cold-stimulus group.Genipin group were treated consecutively with genipin at a dose of 15 mg/kg once a day for 9 days, normal control group were treated with the saline.The mice with cold-stimulus were exposed to 4℃ environment for 5 days.Daily food amount and body weight were measured.Morphological changes were observed in the subscapular region, inguinal region and epididymis around the adipose tissue.The expression of uncoupling protein 1 (UCP1) was determined by real-time PCR and Western blot respectively. RESULTS: The wet weight of white fat in genipin-treated mice was decreased by 16% , and 28% in that of cold-stimulus mice, compared with the normal control group (P＜0.05).After treatments of genipin and cold-stimulus, the color of white adipose tissues was darker, and the size of lipid droplets in adipocytes was smaller, whereas the number was increased.Compared with the normal control group, UCP1 expression was increased obviously in fat tissues, including the subcutaneous and visceral white adipose tissues, and brown adipose tissue after treated with genipin and cold-stimulus (P＜0.05). CONCLUSION: Genipin promoted activation of brown adipose tissue and browning of white adipose tissue by upregulating UCP1 expression, which could contribute to the loss of body weight against obesity.

[Effect of siRNA inhibiting HPV16 E6 gene expression on proliferation and cell cycle of cervical cancer cell line CaSki].

OBJECTIVE: To observe whether human papillomavirus 16 (HPV16) E6-specific small interfering RNAs (siRNAs) can be employed to inhibit the growth of cervical cancer cell line, and to investigate the associated mechanism. METHODS: RNAi was performed using synthetic small interfering RNAs transferred into CaSki cell line by lipofectamine. The cell growth curves, live cell ratio and inhibition ratio of cells were measured by using cell counting. At various time points of post-transfection, the distributions of cell cycle, the expression levels of HPV16 E6, p53, p21 mRNA and proteins were detected by using flow cytometry (FCM) and real-time quantitative reverse transcription-polymerase chain reaction (real-time RT-PCR). RESULTS: The growth inhibition of E6 siRNA to CaSki cells was demonstrated after cells treated with E6 siRNA. No substantial G1 arrest was observed by FCM analysis. For 24 hours after cell transfection, the level of E6 mRNA was decreased by 20. 11 folds compared with control (P < 0.05). However, p53 and p21 mRNA levels appeared unaffected. 48 hours after cell transfection, the expression level of E6 protein was efficiently decreased, but the P53 and P21 protein levels increased in comparison. CONCLUSIONS: The inhibitory effect of HPV16 E6 siRNA to CaSki cell maybe due to specially and efficiently silence E6 mRNA expression, decrease the degradation of wild type P53 protein, and then recover the function activity of P53 protein.

[Inhibiting effect of HPV16 E6 small interfering RNA on the growth of human cervical carcinoma xenotransplanted in nude mice].

OBJECTIVE: To explore the inhibiting effect of human papillomavirus 16 E6-specific small interfering RNA (siRNA) on cervical cancer transplanted in nude mice. METHODS: CaSki cells transfected with HPV16 E6 siRNA were transplanted into nude mice. HPV16 E6 siRNA was injected into the tumor, and the control group was treated with NS. Tumor growth was monitored once every other day. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) was performed to detect apoptosis. The expression of E6 and p53 protein was assessed by immunohistochemistry. The contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured and the liver and kidney were examined by histopathology. RESULTS: Inhibition of tumor growth was demounstrated after treatment with HPV16 E6 siRNA. The volume and weight of tumors were significantly decreased in comparison with those of control group (P < 0.05). Apoptosis occurred more frequently in the experiment group than in the control. The expression of E6 and p53 was down-regulated. The contents of ALT and AST underwent no significant changes, and the histopathology of liver and kidney showed no abnormal changes. CONCLUSION: The growth ability of human cervical cancer xenograft tumors in nude mice can be inhibited by HPV16 E6-specific siRNA, with no toxic side effect on the liver. It may provide an useful method of gene-therapy against cervical cancer.

[Comorbidity of attention deficit hyperactivity disorder in different age group].

OBJECTIVE: To study age distribution of attention deficit hyperactivity disorder (ADHD) comorbidities in a relatively large sample, and exam the hypothesis of bad prognosis for ADHD. METHODS: Using semi-structured clinical diagnosis interview scale, we investigated comorbidities of 1,002 ADHD children and adolescents from psychiatric out-patient clinic, and compared comorbidity frequency in 4 age groups. RESULTS: The comorbidity frequencies of disruptive behavior disorder (DBD), mood disorder, tics disorder and learning disorder (LD) in different age groups of ADHD patients differed significantly (P<0.01). DBD and mood disorder in the age group of 12-14 years (124 cases,51.5%;18 cases,7.5%, respectively) were more than those in the 6-8 years group (160 cases,41.7%;9 cases,2.3%, respectively; P<0.05). And the comorbidity frequencies of the 15-17 years group were significantly higher than those in other age groups (P<0.05). Tics disorder in the 12-14 years group (50 cases,20.8%)was more than those in the two age groups of less than 12 years group (for the 6-8 years group, 51 cases,13.4%;for the 9-11 years group , 42 cases,12.5%; P<0.05).The comorbidity frequency of learning disorder grew up with age. The comorbidity frequency of the 9-11 years group (106 cases,31.5%) was significantly higher than that of the 6-8 years group (80 cases,20.9%; P<0.01); and those of both age groups more than 12 years (for the 12-14 years group, 164 cases,68.0%;for the 15-17 years group, 28 cases,70.0%) were higher than those of the two age groups of less than 12 years (P<0.01). ADHD patients without any comorbidities in the two age groups more than 12 years (for the 12-14 years group, 29 cases,12.0%;for the 15-17 years group, 4 cases,10.0%)were less than those in the two age groups of less than 12 years (for the 6-8 years group, 126 cases,32.8%;for the 9-11 years group, 110 cases,32.6%; P<0.01). These characteristics maintained in boys. CONCLUSION: Many comorbid psychiatric disorders increase as the ADHD children become adolescents. The situation is severer in late adolescence. Therefore, ADHD children should accept appropriate treatment as early as possible, and adopt the second degree prevention actively.

[Association analysis of dopamine D4 receptor gene polymorphism and attention deficit hyperactivity disorder with/without disruptive behavior disorder].

OBJECTIVE: To examine the role of dopamine D4 receptor gene (DRD4) in attention deficit hyperactivity disorder (ADHD) with/without disruptive behavior disorder (DBD) by focusing on a-521C/T SNP within the promoter region of this gene. METHODS: A total of 401 DSM-IV ADHD children (including 284 trios) of Chinese Han descent were genotyped. Chi-square test and the transmission disequilibrium test (TDT) were used to test for associations in ADHD with and without DBD respectively. RESULTS: In the comparison of ADHD with (n=143) and without (n=258) DBD, the -521T allele ( chi(2) = 6.778, P= 0.009, OR= 1.485) and the TT genotype (chi(2) = 6.292,P=0.012,OR=1.729) showed higher frequency in children with ADHD and DBD simultaneously. For family based analysis, T allele of the -521C/T polymorphism was preferentially transmitted to ADHD children with comorbid DBD (n=100, chi(2) = 3.868,P=0.049), whereas no significant distortion was found in the transmission of the tested variant for ADHD without DBD (n=184, chi(2) = 0.223, P=0.637). CONCLUSION: Our findings suggest that the -521C/T SNP of DRD4 may contribute to the predisposition to ADHD with comorbid DBD. This study supports for the hypothesis that ADHD with comorbid DBD may be influenced by greater genetic effect compared to ADHD alone.

[Experimental study on inhibition of cervical carcinoma siha cell proliferation by siRNA expression vector].

OBJECTIVE: To observe the effect of HPV16E7 specific expression vector on cell proliferation in cervical carcinoma SiHa cells. METHODS: The HPV16E7 siRNA expression vector and empty expression vector were transfected into SiHa cells by liposome. The effects on E7 mRNA and E7 protein expression, cell cycle phase and cell growth rate were examined respectively by real-time RT-PCR, FCM and MTT assay. RESULTS: The HPV16E7 siRNA expression vector significantly inhibited the expression levels of E7 mRNA and E7 protein, the inhibition rates being 92.15% and 84.30% respectively. It also inhibited the transition from G, phase to S phase and the growth of SiHa cell line. CONCLUSION: HPV16E7 specific siRNA expression vector could specifically and efficiently inhibit the expression of E7 gene and hence it could regulate cell cycle and inhibit cell proliferation in cervical carcinoma SiHa cells. siRNA expression vector

Electroacupuncture suppresses expression of gastric ghrelin and hypothalamic NPY in chronic food restricted rats.

Electroacupuncture (EA) has been reported to reduce body weight in overweight subjects in clinical practice, as well as in rats and mice with diet-induced obesity. In the present study, this effect of EA was tested in lean rats subjected to long-term food restriction (FR, food was offered only 1 h/day). Two hertz EA administered once every other day produced a further reduction in body weight in FR rats. Exploration of the mechanism involved revealed significant downregulation of the orexigenic peptides: ghrelin in the stomach, and neuropeptide Y (NPY) but not Agouti-related peptide (AgRP) in the hypothalamus, which was in line with the reduction in food intake in rats receiving EA stimulation as compared with those receiving restraint only. Uncoupling protein 3 (UCP3), involved in accelerating energy expenditure, was not significantly altered. These results suggest that the EA-induced body weight reduction was due mainly to a decrease in food intake rather than an increase in energy expenditure. A reduction in the orexigenic peptides ghrelin and NPY may be involved in the underlying mechanism.

Salidroside alleviates oxidative stress in the liver with non- alcoholic steatohepatitis in rats.

BACKGROUND: Nonalcoholic steatohepatitis (NASH) is characterized by fat accumulation in the hepatocyte, inflammation, liver cell injury, and varying degrees of fibrosis, and can lead to oxidative stress in liver. Here, we investigated whether Salidroside, a natural phenolic antioxidant product, can protect rat from liver injury during NASH. METHODS: NASH model was established by feeding the male SD rats with high-fat and high-cholesterol diet for 14 weeks. Four groups of male SD rats including, normal diet control group, NASH model group, and Salidroside treatment group with150mg/kg and 300 mg/kg respectively, were studied. Salidroside was given by oral administration to NASH in rats from 9 weeks to 14 weeks. At the end of 14 weeks, liver and serum were harvested, and the liver injury, oxidative stress and histological features were evaluated. RESULTS: NASH rats exhibited significant increases in the following parameters as compared to normal diet control rats: fat droplets with foci of inflammatory cell infiltration in the liver. ALT, AST in serum and TG, TC in hepatocyte elevated. Oxidative responsive genes including CYP2E1 and Nox2 increased. Additionally, NASH model decreased antioxidant enzymes SOD, GSH, GPX, and CAT in the liver due to their rapid depletion after battling against oxidative stress. Compared to NASH model group, treatment rats with Salidroside effectively reduced lipid accumulation, inhibited liver injury in a does-dependent manner. Salidroside treatment restored antioxidant enzyme levels, inhibited expression of CYP2E1 and Nox2 mRNA in liver, which prevented the initial step of generating free radicals from NASH. CONCLUSION: The data presented here show that oral administration of Salidroside prevented liver injury in the NASH model, likely through exerting antioxidant actions to suppress oxidative stress and the free radical-generating CYP2E1 enzyme, Nox2 in liver.