Objective To observe the effect of piperine (PA) on experimental gallstone formation in the gallbladder of C57BL/6 mice. Methods 3 dietary groups of C57BL/6 with 10 mice each group were allocated as control (normal mice chow),lithogenic (1% cholesterol diet) and PA (1% cholesterol diet + PA 30 mg/kg body weight) group respectively for 4 weeks. The expression of Scp2 gene in liver tissue was measured by RT-PCR and the bile lipid contents was measured chemically,the cholesterol saturation index (CSI) was calculated by Carey’s method. Results Cholesterol crystals and stones were found in 10/10 and 9/10 gallbladders respectively in lithogenic group,but not found in control and PA groups. Comparing with the lithogenic group,the expression of Scp2 gene in liver tissue and CSI in the gallbladder bile were significantly decreased in PA group. Conclusions PA inhibit the experimental gallstone formation induced by high cholesterol feeding in C57BL/6 mice,with simultaneous decreasing of both the Scp2 gene expression in the liver tissue and the CSI value in the gallbladder bile. The further study of the preventive effect of gallstong formation of PA,whether or not related to the above results,is strongly suggested.

Objective To investigate the relationship between genetic polymorphisms of Nogo gene and nasopharyngeal carcinoma in Zhuang ethnic group in Guangxi Province.Methods Nogo gene polymorphisms were analyzed between patients with nasopharyngeal carcinoma and healthy people as a control in Zhuang ethnic group in Guangxi province using PCR-SBE.Their genotype and allele frequency distribution were compared between case and control group.The frequencies of haplotypes were analyzed with SHEsis software between these two sites in different groups.Results There were no differences between the patients and controls in the genotype or allele frequencies of Nogo gene rs 17046518 site (P ＞ 0.05).But the frequency distribution of T allele was significantly different (P =0.003) in the rs12464595 site.The result of haplotypes analysis showed that GA haplotype and CA haplotype were significantly different between cases and control (P =0.045,P =0.002).Conclusion The CG haplotype,CA haplotype and T allele of Nogo gene rs12464595 site increase the risk of nasopharyngeal carcinoma among Zhuang ethnic group in Guangxi.

Objective:To investigate the association between LPHN3 andattention-deficit/hyperactivity disor-der(ADHD)in Chinese Han children.Methods:Based on the Diagnostic and Statistical Manual of Mental Disor-ders,Fourth Edition (DSM-IV)diagnosis criteria,921 normal controls and 1052 ADHD children were included in the study.The Clinical Diagnostic Interview Scale (CDIS )was used to assess symptoms and ADHD sub-types.ADHD was divided into three subtypes,namely ADHD inattentive type (ADHD-I),ADHD hyperactive-im-pulsive type (ADHD-HI),and ADHD combined type (ADHD-C).The ADHD rating-scale was used to assess ADHD symptoms.Including inattentive symptoms,hyperactive symptoms,impulsive symptoms and ADHD total symptoms.Three single nucleotide polymorphisms (SNPs)of LPHN3 were genotyped.Case-control studies were conducted to investigate the association of each SNP with the ADHD and the subgroups using chi-square test.Results:rs11131347 was associated with ADHD P <0.05,OR =0.86(0.76 -0.98)],but the difference didn't survive significance after corrections,The frequency of minor allele C in cases and control was0.409 vs.0.445.For different genders,rs11131347 was significantly associated with ADHD boys [P <0.05,OR =0.82(0.71 -0.96)], The frequency of minor allele C in cases and control was0.402 vs.0.449.For different subtypes,rs11131347 was associated with ADHD-C [P <0.05,OR =0.85(0.74 -0.98)]and ADHD-C boys[P <0.05,OR =0.82(0.70 -0.97)],but none of the difference survived significance after corrections,The frequencies of minor allele C in cases and control were respectively 0.407 vs.0.445 and 0.401 vs.0.449.Genotypes distribution analysis indicated that rs11131347 was associated with ADHD in general (recessive model,P <0.05),ADHD boys (additive model,P <0.05;dominant model,P <0.05;recessive model,P <0.05),ADHD-C(recessive model,P <0.05)and ADHD-C boys (dominant model,P <0.05 ),however,none of the difference survived significance after correc-tions.rs11131347 was nominal associated with impulsive scores(P <0.05).Conclusion:These findings suggest that the polymorphism of LPHN3 is probably involved in the pathological mechanisms of ADHD and its core symptoms of impulsivity.

AIM: To investigate the effect of caveolin - 1 and phosphorylation of ERK1/2 on 17β - estradiol ( E_2 ) induced inhibition of vascular smooth muscle cells ( VSMCs ). METHODS: The proliferation in cultured VSMCs was determined by using [~3H ] - thymidine incorporation. The expressions of caveolin - 1, MKP -1 and ERK1/2 phosphorylation were measured by Western blotting. The expression of caveolin - 1 mRNA was measured by RT - PCR. RESULTS: Exposed to fetal calf serum ( FCS) for 24 h, the increase in proliferation of VSMCs was detected by [~3H] -thymidine incorporation. Pretreatment with various concentrations of E_2 for 24 h inhibited VSMC proliferation induced by FCS. The results of Western blotting and RT - PCR showed that pretreated with 17β - estradiol for 24 h reserved the decrease in caveolin - 1 induced by FCS. Western blotting results further proved that the expression of MKP - 1 was significantly increased and the expression of ERK1/2 phosphorylation was decreased after incubated with 17β - estradiol. CONCLUSION: 17β -estradiol increases caveolin - 1 and MKP - 1 expressions, and decreases ERK1/2 phosphorylation, leading to the inhibition of VSMC proliferation.

Objective To explore the role of human papillomavirus (HPV) in squamous cell carcinoma of the larynx.Methods A cohort of 152 laryngeal squamous cell carcinoma (LSCC) patients,40 premalignant lesions of the larynx,and 40 vocal fold polyp patients was evaluated for the expression of HPV16 E7 protein by immunohistochemistry (IHC).E7 protein expression was correlated with clinicopathological features and outcomes.Results The expression of E7 protein in vocal fold polyp was negative,and it's positive expression in precancerous lesions and laryngeaI carcinomas was 2.5％,and 32.9％,respectively.There were no other significant clinical or demographic differences between pl6-positive and-negative cases.There was no difference in overall survival (OS) between E7-positive and-negative patients with 5-year survival of 79％ and 66％ for p16 positive and negative patients,respectively (P =0.581).There was no statistically significant difference in failure-free survival (FFS) with 5-year FFS of 57.8％ and 42.3％ for p16 positive and negative patients,respectively (P =0.579).Conclusions E7 expression is infrequent in laryngeal carcinomas.There are no statistically significant correlations between E7 IHC status and OS or FFS.

BACKGROUND: Central oxytocin (OT) may be a neurotransmitter or neuromodulator and play an important role in learning and memory, sexual behaviour, pain modulation and opiate tolerance and dependence. To research the interactions between oxytocinergic and opioidergic system in hippocampus has some significance.OBJECTIVE: To investigate the effects of OT administered intracerebroventricularly on evoked discharge of left dorsal hippocampal CA1 neurons in rats and the possible interactions between oxytocinergic and opioidergic system.DESIGN: A randomised controlled study.SETTING: Department of Physiology of Guangdong Medical College; Department of Physiology and Pathology of Medical College of Wuhan University.MATERIALS: The study was conducted in the Physiology Department of Medical College of Wuhan University from September 2002 to September 2003. A total of 36 male Sprague-Dawley rats were selected and randomly divided into six groups: control (NS), OT groups (0.2 mg/L, 2 mg/L and 20 mg/L), [d (CH2)5-OVT]+OT (2 mg/L), naloxone+OT (2 mg/L), with 6 rats in each group.METHODS: Single-unit recording was performed with glass microelectrode. The glass microelectrode was inserted by a micromanipulator into hippocampal CA1. The electrical activity was amplified by a microelectrode amplifier and then recorded by the biological experimental system,monitored at the same time with oscilloscope. When recording the neural discharge, electrical stimulation of the sciatic nerves was performed once 5minutes through a double stainless electrode. 5 μL oxytocin in dosage of 0.2, 2 and 20 mg/L were injected slowly into lateral ventricle via microlitre syringe. [d(CH2)5-OVT]+OT (2 mg/L) group: 2.5 μL [d(CH2)5-OVT](80 mg/L) was injected into lateral ventricle and then 2.5 μL oxytocin (2 mg/L). Naloxone+OT (2 mg/L) group: 2.5 μL naloxone (400 mg/L) was injected into lateral ventricle and then 2.5 μL oxytocin (2 mg/L). According to frequency of discharge, effect of oxytocin at various dosages on discharge induced by neurons in hippocampal CA1 area and [d (CH2)5-OVT]and naloxone on oxytocin was assayed. MAIN OUTCOME MEASURE: Changes of discharge frequency after stimulation.RESULTS: Data of totally 36 rats were entered the final analysis. ① OT (0.2 mg/L, 2 mg/L and 20 mg/L) administered by intracerebroventricularly could decrease the evoked discharge of hippocampal CA1 neurons in a dose-dependent manner. ② The inhibitory effects of OT (2 mg/L) could be blocked by pretreated intracerebroventricularly injection of [d (CH2)5-OVT](80 mg/L, 2.5 μL). ③ Intracerebroventricular injection of naloxone (400 mg/L, 2.5 μL) could attenuate the effects of OT (2 mg/L) significantly.CONCLUSION: OT can inhibit the electrical activities of hippocampal CA1 neurons to external electrical signal through activating the oxytocin receptor. Moreover, central opioid receptor is involving in the inhibitory effects of OT.

Objective To observe the expression of apoptosis and invasion genes in different tissues of laryngeal lesions,and to detect the content of proliferation related protein and proliferation inhibition gene.Methods In our hospital,the clinical diagnosis of laryngeal carcinoma,precancerous lesions and vocal polyps in 88 cases of laryngeal carcinoma,surgical excision of lesion specimens cut from the amount of Pro apoptotic genes and promote invasion and proliferation related protein,gene the content of tissue proliferation inhibition gene detection.Results The laryngeal carcinoma group of Pro apoptotic genes nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) and large tumor suppressor gene 1 (LATS1) were all significantly lower than lesion group and polyp group (P ＜ 0.05).Laryngeal carcinoma invasion promoting gene tumour necrosis factor receptor associated factor 6 (TRAF6),and cellular FADD-like interleukin1βconverting enzyme inhibitory protein (c-FLIP) content were significantly higher than that of laryngeal pre cancerous lesions and vocal polyps (P ＜ 0.05).The proliferation related protein Cyclin-dependent kinase 6 (CDK6) and E2F1 in laryngeal carcinoma were significantly lower than that in the lesion and polyp group,and cyclin D1 (CCND1),Bmi-1,and Livin contents were significantly higher than that in the lesion group and the polyp group (P ＜ 0.05).The levels of differentiated embryo-chondrocyte expressed gene 1 (DEC-1),IκB kinase 16(IKK16),large tumor suppressor gene 1 (LAST-1),receptor-interacting proteins-1-(RIP-1),and c-myc promoter binding protein-1 (MBP-1) in the tissue of laryngeal carcinoma were significantly lower than those in the lesion group and polyp group (P ＜ 0.05).Conclusions The laryngeal carcinoma invasion promoting gene was higher than that in other tissues,inhibition of gene content was lower than that of other tissues proliferation related protein gene,apoptosis and proliferation,apoptosis and invasion of lesions in gene expression level,proliferation related protein and proliferation inhibition of gene content and laryngeal squamous cell carcinoma were related to laryngeal disease prevention significance of detection.

Objective To investigate the changes of the activity of blood coagulation and fibrinolysis in fracture patients,and to research the relations between the activity and thrombossis. Methods This reser ch chose 60 patients in the department of orthopaedic trauma in our hospital and 10 normal control.Each patient enderwent laboratory analysis on preoperation?post operation 24 hours?post operation 4 days.The molecule markers of coagulation and fibrinolysis TAT(thrombin/antithrombin Ⅲ complex)?F1+2(prothrombin fragment F1+2)?PAP(Plasmin/? 2-antiplasmin complex) were quantitative measured by enzyme immunoassay. Result Compare with normal control,F1+2 and TAT levels increase obviously(P0.05); FG levels increase obviously (P0.05) .Conclusion It has a high level of the activity of blood coagulation and has no obviously change of the activity of fibrinolysis after trauma injury,it indicates high coagulation activity is correlated with thrombossis.

AIM:To investigate the effect of caveolin-1 and phosphorylation of ERK1/2 on 17?-estradiol (E2) induced inhibition of vascular smooth muscle cells (VSMCs). METHODS:The proliferation in cultured VSMCs was determined by using [3H]-thymidine incorporation. The expressions of caveolin-1,MKP-1 and ERK1/2 phosphorylation were measured by Western blotting. The expression of caveolin-1 mRNA was measured by RT-PCR. RESULTS:Exposed to fetal calf serum (FCS) for 24 h,the increase in proliferation of VSMCs was detected by [3H]-thymidine incorporation. Pretreatment with various concentrations of E2 for 24 h inhibited VSMC proliferation induced by FCS. The results of Western blotting and RT-PCR showed that pretreated with 17?-estradiol for 24 h reserved the decrease in caveolin-1 induced by FCS. Western blotting results further proved that the expression of MKP-1 was significantly increased and the expression of ERK1/2 phosphorylation was decreased after incubated with 17?-estradiol. CONCLUSION:17?-estradiol increases caveolin-1 and MKP-1 expressions,and decreases ERK1/2 phosphorylation,leading to the inhibition of VSMC proliferation.

Objective To explore the impacts of catechol-O-Methyltransferase (COMT) gene polymorphisms on the outcomes of methylphenidate treatment for attention deficit hyperactivity disorder (ADHD) in children. Methods One hundred seventy-seven ADHD children of Chinese Han descent received open-labelled dose titration with methylpheni?date to achieve optimal response in 2~4 weeks. The behavior changes were evaluated by using ADHD diagnostic scale (parent version) before and after treatment. COMT gene rs4680 (Val158Met) and rs165599 were genotyped using fluores?cent real-time PCR. The genotype distribution and treatment outcomes including remission, response and non-response were analyzed. Results The treatment response differed significantly among patients with different genotypes of rs4680 (P<0.01). Thirty-eight (40.9%) patients with G/G genotype had remission and 19 (20.4%) had good response;Thirty-five (48.6%) patients with G/A genotype had remission and 16 (22.2%) had good response. In contrast, only one patients with A/A genotype had remission and none of them had good response. The changes of the ADHD diagnostic scale total scores, inattention scores and hyperactivity impulsive scores were statistically different among different genotypes of rs4680 (P<0.05). The hyperactivity impulsive scores were significantly higher in the G allele carrier than A allele carrier (P=0.04). The treatment response was not significantly different among different rs165599 genotypes (P>0.05). Conclu?sion COMT gene rs4680 (Val158Met) polymorphism is associated with methylphenidate response in a Han Chinese popu?lation. Patients with G allele is more likely to benefit from methylphenidate treatment in comparison with A/A genotype.