Objective To explore the correlation between aspirin resistance and COX-1, P2Y1, GPIa and GPIIIa gene polymorphism. Methods 35 case with aspirin resistant were selected and were given aspirin enteric coated tablets for oral treatment, 14 days.After the determination COX-1, P2Y1 and GPIa and GpIIIa gene polymorphism of each patient were analysis.Results The genotype AA of COX1 (A-842G) locus and CC of COX1 (C50T) locus was higher than other genotype (P<0.05);AG of P2Y1 (A1622G) locus genotype and CC of P2Y1 (C893T) locus genotype was higher than other genotype (P<0.05);CT of GPIa (C807T) locus genotype and GA of GPIa (G873A) locus genotype was higher than other genotype (P<0.05);PLA1/A1 of GPⅢa(T1565C)locus genotype was higher than other genotype (P<0.05).Conclusion P2Y1 (C893T), GPIa (C807T) allele is associated with aspirin resistance,With COX-1 (A-842G, C50T), GPIa (G873A), GP III A (T1565C), P2Y1 (A1622G) allele is more likely to induce the occurrence of aspirin resistance.

Objective To clone the coding dense granules protein 8(GRA8) from Toxoplasma gondii RH isolate for the potential use in the development of DNA vaccination. Methods Amplifing gene fragment coding GRA8 from the genomic DNA of Toxoplasma gondii RH isolate by means of ploymerase chain reaction (PCR), the gene is inserted into cloning vector pUC-19 digesting with restrictive enzymes and linking react ions. The positive colon is screed on LB plates containing ampicilline and IPTG, Xgal identified by blue-white and restrictive enzyme digestion. The inserted GR A8 gene was recombined with pcDNA3.1(+) eukaryotic expression vector by digestion with restrictive enzyme and linking reactions. The positive coloe is screene d o n LB plates containing ampicillin and identified by restrictive enzymes and link ing reactions. Results The GRA8 gene with about 804 base is specifically amplified from genomic DNA of Toxoplasma GRA8/RH and pcDNA3.1(+)-GRA8 recombinant is successfull y constructed. The sequencing results showed that GRA8 gene of isolate RH and R H from genebank shares quite high homology. Conclusion The gene encoding GRA8 is amplified from genomic DNA of Toxoplasma gene isolate GRA8/RH and pcDNA3.1 (+)-GRA8 recombinant is successfully constructed.

To compare the effects of four different intensive insulin therapies on blood glucose control and vascular endothelial function in newly-diagnosed type 2 diabetes.Patients were randomly divided to accept pre-meal insulin aspart 30 or pre-meal insulin aspart and glargine at bedtime or pre-meal Novolin-R and NPH at bedtime or continuous subcutaneous insulin aspart infusion.Capillary blood glucose determination and continuous glucose monitoring system were carried out,therapeutic time and total insulin dosage were recorded.Ultrasound was used to evaluate the vascular endothelial function.Glucose level,incidence of low glucose,potency ratio of the four groups were similar( P＞0.05 ) ; FMD and NMD were not significantly improved ( P =0.718,P =0.065 ).The short-term efficacy and safety of the four groups are similar.The short-term intensive insulin therapy has no obvious effect on vascular endothelial function.

Objective To investigate the status of the non-psychiatrist' s diagnostic and therapeutic ability of depression/anxiety disorders in 57 General Hospitals.Methods 1 152 non psychiatric clinicians in 57 general hospitals were surveyed.A custom-made questionnaire included the training of mental health-related knowledge which the general hospital physicians received and typical anxiety/depression case analysis.Results Among 1 521 non-psychiatric clinicians,596 (51.74％) clinicians participated the training of psychiatry,562 (48.78％) participated the training of medical psychology and 230(20.97％) clinicians participated the training of Healthy Psychology.In professional setting,59 (5.12％) clinicians participated the training of psychotherapy,255 (22.14％) clinicians had attended related academic symposiums.80(6.94％) clinicians believed that they understand the clinical display of anxiety/depression disorders,52 (4.51％) clinicians expressed the understanding of diagnostic criteria of anxiety/depression disorders and its treatments,while 44(3.82％) clinicians only possessed the knowledge of anxiety/depression disorder treatment.In the typical case analysis,it revealed that 794 (68.89％) clinicians made accurate diagnosis,458(57.68％)clinicians made a choice of medical treatment,764(96.22％) clinicians chose psychotherapy,29 (3.65％) clinicians applied physical therapy,while 438 (55.16％) clinicians combined drug therapy with one or more other therapeutic methods.Binary logistic regression analysis showed that the age(P=-0.093,Exp(B)=0.911)and work experience(P=-0.002,Exp(B) =1.080)significantly contribute to the diagnostic accuracy of non-psychiatrists.Conclusion The non-therapeutic psychiatric clinicians in general hospitals have certain basic knowledge of depression/anxiety disorders with lower level of diagnosis and treatment diagnosis and treatment.And there is bigger difference among different hospitals.

AIM:To investigate atrial natriuretic peptide(ANP) circadian in the encapsulated human ANP(hANP) cDNA transfected cells,to alter the ANP circadian by artificial control to achieve the objective of effectively treat hypertension or congestive heart failure(CHF). METHODS:ANP cDNA was transfected into Chinese hamster ovary(CHO) cells,which were encapsulated in polycarprolactone(PCL) tubes.The longterm survival of transfected CHO cells and the levels of ANP secreted were detected.Circadian rhythm of ANP secreted by encapsulated transfected cells was also studied,which was regulated by melatonin. RESULTS:During culturing,the ANP level secreted by transfected CHO cells in 2 mL of culture medium within 24 hours could reach 210.3 ng/L in a 20 mm-long and 2 mm-diameter PCL tube.The section of ANP displayed a circadian variation:higher in daytime,but lower at night.The acrophase of circadian rhythm was 4:15 but could be shifted to 7:55 after melatonin management. CONCLUSION:ANP cDNA transfected CHO cells that encapsulated into PCL tube can secret ANP,which might be suitable for the future implantation into human body.Our research provides a new approach in the treatment of hypertension and CHF by ANP.

Aim To investigate the effects of fluoxe-tine on the changes of of protein levels of GLT-1 in pre-frontal cortex in rat depression model, and to further explore the molecular mechanism of antidepressant ac-tion of fluoxetine. Methods Sixty male SD rats were randomly assigned into three groups: control group, chronic unpredictable stress ( CUS) group, and CUS+fluoxetine group. The rats of CUS group and CUS+flu-oxetine group were subjected to CUS for 2 sessions per day for 35 days. Then, the rats of the CUS+fluoxetine group were given fluoxetine for 28 days. Behavioral changes were assessed by the sucrose preference and open field tests. The GLT-1 protein levels in the pre-frontal cortex were detected by immunohistochemistry and Western blot analysis at the end of the fluoxetine treatment. Results ( 1 ) Compared with the control group,sucrose preference, total traveling distance, ve-locity and frequencies of rearing were reduced in the CUS group ( P < 0. 01 ) . These behavioral changes could be reversed after 28 day fluoxetine treatment. (2 ) Immunohistochemistry assay indicated weak im-munoreactivity for GLT-1 in the prefrontal cortex of CUS group ( versus the control rats: P <0. 01 ); the immunoreactivity for GLT-1 of the fluoxetine-treated rats was significantly up-regulated compared with the CUS group rats ( P<0. 01 ) . ( 3 ) Western blot analy-sis indicated significant reductions of GLT-1 in the pre-frontal cortex of CUS group ( versus the control rats:P<0. 01 ) , and chronic fluoxetine treatment reversed the CUS-induced decrease in GLT-1 levels ( P <0. 01 ) . Conclusions Chronic unpredictable stress ( CUS ) could down-regulate the GLT-1 protein levels in the prefrontal cortex, which is reversed by fluoxe-tine. These results further support the notion that en-hanced expression of the GLT-1 protein could be mo-lecular mechanism of fluoxetine antidepressant effect.

Objective To investigate the effects of different duration of fluoxetine and escitalopram administration on depressive-like behavior and hippocampal BDNF expression in adult rats.Methods Ninety-six SD rats were randomly divided into twelve groups: (1)M 1 group: normal control for one week, (2)M2 group: CUMS +saline for one week, (3)M3 group: CUMS+fluoxetine for one week, (4) M4 group: CUMS+escitalopram for one week, (5) M5 group : normal control for two weeks, (6) M6 group : CUMS+ saline for two weeks, (7) M7 group : CUMS+fluoxetine for two weeks, (8)M8 group: CUMS+escitalopram for two weeks, (9)M9 group: normal control for three weeks,(10) M10 group: CUMS+saline for three weeks, (11)M11 group: CUMS+fluoxetine for three weeks, (12) M8 group: CUMS+escitalopram for three weeks.After CUMS procedures,rats in M2 group,M6 group and M10 group were injected with saline, M3 group, M7 group and M11 group were injected with fluoxetine, and rats in M4 group,M8 group,M12 group were injected with escitalopram.After one week of intervention,the openfield test and 1％ sucrose preference test were performed to evaluate depression-like behaviors in rats of M1 group, M2 group,M3 group and M4 group.After behavior test,rats were sacrificed and the hippocampi were isolated.The expression of BDNFmRNA was detected by using real-time quantitative PCR.After two weeks of intervention, rats in M5 group,M6 group, M7 group and M8 group underwent the same behavioral.After three weeks of intervention, rats in M9 group, M10 group, M11 group and M12 group underwent the same behavioral test.Results In the open-field test,total distance travelled in 10 minutes was significant difference among the following groups: M1 group ((3925.70±322.32) cm) vs M3 group ((1841.85±786.33) cm) ,M6 group ((1820.31±296.00) cm) vs M8 group ((4002.72± 1447.19) cm), M10 ((1961.66±919.16) cm) group vs M11 group ((3741.72± 1064.46)cm) ,M10 group ((1961.66±919.16) cm) vs M12 group ((4280.43±1187.05) cm).In the 1％ sucrose preference test,the difference of sucrose preference consumption was statistically significant (P＜0.05) among the following groups: M2 group ((56.23±7.49)％) vs M4 group ((70.55±4.96)％), M6 group ((60.22±8.81)％) vs M8 group ((75.08±4.15)％) ,M10 group ((60.26±7.20)％) vs M11 group ((73.88±7.73)％) ,M10 group ((60.26 ± ±7.20)％) vs M12 group ((73.52±7.58)％).The expression level of BDNF was significant difference among these groups: M2 group (0.66±0.14) vs M4 group (1.15±0.20) ,M10 group (0.90±0.15) vs M11 group (1.22± 0.09) ,M10 group (0.90±0.15) vs M12 group (1.48±0.20).Conclusion Both of fluoxetine and escitalopram can improve depression-like behaviors in rats and significantly increase the expression of the hippocampal BDNFmRNA.Compared with fluoxetine,escitalopram has a shorter onset time in the treatment of depression.It may be related with a rapid increase of the expression of BDNF mRNA.

Objective To investigate the characteristics of the event related potential(ERP) P300 and analyze brain network connections in patients with first-episode depressions.Methods P300 auditory oddball task were administrated on twenty-nine patients and twenty-five healthy controls.The P300 amplitude and latency of two groups were compared,and the brain network connectivity of the two groups were analyzed using Granger's Causality analysis.Results The P300 amplitude in depression group were significantly different from those in control group (C3 of the central regions(15.77±7.35) μV vs (20.90±7.82)μV;C4 of the central regions(16.98±7.21) μV vs (22.11±7.50) μV;P3 of the parietal regions(15.65±6.92) μV vs (19.49±5.73) μV;P4 of the parietal(16.35± 6.46) μV vs P4(19.72±5.18) μV;P=0.009,P=0.007,P=0.017,P=0.024 respectively).However,the P300 latency had no significant difference comparing to the controls(P＞0.05).The results also showed that patients had more connections in the brain network.Conclusion As an effective evaluation index,ERP P300 can play an important role in clinical diagnosis of depression.Patients suffering from depression have significant cognition function deficit.

Objective:To construct a short hairpin RNA(shRNA) eukaryotic expression vector of the survivin gene,and investigate its inhibitory effect on the survivin expression in human cervical cancer cells.Methods:We designed and synthesized 2 pairs of survivin-specific small interfering RNA primers(s1 and s2),cloned them into the eukaryotic expression vector pSilencer 2.1-U6 neo by DNA recombined techniques after annealing connection reaction,and transfected them respectively into human cervical cancer cell line HeLa using LipofectAMINE2000 after identificationby restrict endonuclease digestion and DNA sequencing.Then we selected the positive clones by G418 and detected the expression of survivin mRNA by semi-quantitative RT-PCR and that of the survivin protein by Western blot.Results:The survivin shRNA eukaryotic expression vectors pSilencer2.1-s1 and pSilencer2.1-s2 were successfully constructed.Positive clones were obtained by screening with G418 for 24 days,the survivin expression in HeLa cells decreased in different degrees after transfected with pSilencer2.1-s1 and pSilencer2.1-s2,and the latter showed a better interference efficiency.Conclusion:The shRNA eukaryotic expression vector of the survivin gene we constructed,with its improved interfering efficiency,has paved the way for further research on its role in regulating the biological behavior of cervical cancer cells.

The glutamate transporter EAAT 2 ( rodent nomencla-ture GLT-1:glutamate transporter 1), which is a predominantly astroglial glutamate transporter in the hippocampus and the pre-frontal cortex , is responsible for the majority of extracellular glu-tamate uptake .The glutamate transporter EAAT 2 can decrease the high levels of glutamate in the synaptic cleft , avoiding gluta-matergic excitotoxicity to damage the glial cells and neurons . Currently, the transporter EAAT2 has become a research hotspot of depression .This article aims to summarize roles of glutamate transporter EAAT2 in the occurrence and treatment of depres-sion.