Objective To clone the telomere-associated protein T-STAR and study the relationship between T-STAR and the telomerase catalyzed subunit hTERT in mammalian cells. Method The expression vector pGBKT7-hTERT was constructed and acted as a decoy in cDNA library screened by yeast two-hybrid technology. Recombinant vectors pVP16-T-STAR and pM-hTERT were constructed and co-transfected with report gene CAT into SGC-7901 cells with liposome. pM-53+pVP16-T and pM3-VP16 were introduced as positive controls, pM-53+pVP16-CP as negative control, and pM-hTERT+pVP16, pM+pVP16-T-STAR and pM+pVP16 as crossing negative controls. The expression of CAT was assayed by ELISA. Results The eukaryotic expression vector pGBKT7-hTERT was successfully constructed. One positive clone achieved by cDNA library screening was sequenced and compared with the isogenous sequences in GenBank by Blast software via Internet. As a result, T-STAR, a recorded cDNA sequence was obtained. The recombinant vectors of pVP16-T-STAR and pM-hTERT were constructed successfully and co-transformed into gastric cancer cells SGC-7901. The OD value of reported gene CAT was 0.258, which was significantly higher than that of the negative and crossing negative controls (0.002-0.015). It revealed that T-STAR interacted with hTERT in the mammalian cells. Conclusions T-STAR interacts with hTERT in the gastric cancer cells. T-STAR may be a new member of telomere-associated protein, and it participates in the regulation of telomerase through hTERT.

Objective:To study the effects of Naoling tablet on Aristolochia acid A from Aristolochia fangchi,preliminary explore the deintoxication of Aristolochia fangchi.Methods:Absorption value of Aristolochhia aicd A was determined by HPLC after Aristolochi fangchi decocted with Naoling tablet,the change of content of Aristolochia acid A was observed .Results:the content of Aristolochia acid A from Aristolochia fangchi was markedly decreased after Naoling tablet was added.Conclusion:Naoling tablet can markedly decreased the content of Aristolochhia aicd A from Aristolochia fangchi.

Objective To study the effects of Ziyin Bushen Pill on Aristolochia acid A in Aristolochia fangchi,and preliminary study on reducing poisonous effects of Ziyin Bushen Pill to Aristolochia fangchi.Methods Absorption of Aristolochhia acid A in the mingled decoction of Aristolochia fangchi and Ziyin Bushen Pill by HPLC,and the change of Aristolochia acid A was observed.Results Aristolochia acid A content of Aristolochia fangchi was markedly decreased after Ziyin Bushen Pill was added.Conclusion Ziyin Bushen Pill can markedly decreased Aristolochhia acid A content of Aristolochia fangchi.

Objective To analyze the characteristics,risk and induced factors of intussusception in Peutz-Jeghers syndrome (PJS).Methods From March 2nd 2005 to May 25th 2013,a total of 130 patients with PJS were selected.The clinical data of patients were collected,which included gender,age,the diagnostic age of PJS,family history,the diagnostic age of intussusception,clinical features,location,treatment and the maximum diameter of polyps which caused intussusception.Kaplan-Meier was performed to analyze the cumulative risk of intussusception.Results Among 130 patients with PJS,90 cases had intussusception.The age when first time diagnosed was from four to 33,median age was 16.The cumulative risks of intussusception at 10,15,20,25 and 30 years were 12.308％ (16/130),31.538％(41/130),51.538％(67/130),64.615％(84/130) and 66.923％(87/130).There was no significant difference in the cumulative risk of intussusception between male and female; with family history and no family history (both P＞0.05).A total of 131 intussusception happened in 90 patients,of which diagnosed by surgery,imaging examination and history reviewer was 125,four and two times,respectively.The initial symptom of 111 times of intussusception was acute abdomen and 15 were abdominal pain and vomiting.The left five intussusception was found by regular:examination.One hundred and fifteen intussusception was in small intestine and 16 in colon.There was 127,three and one time treated with surgery,conservative treatment,endoscopic therapy (dual airbags intestinal endoscopic polypectomy),respectively.The maximum diameter of polyps which caused intussusception was from 15 to 70 mm,average 40 mm.Conclusions Intussusception of patients with PJS is at young age and with a high cumulative risk.Intussusception is generally caused by diameter over 15 mm polyps.

AIM To observe the effect of intra- venous injection of erythromycin (EM) on interdigestive migrating motor complex (MMC) and postprandial gastrointestinal contraction in conscious dogs, and to study its possible mechanism. METHODS Gastrointestinal contractile activity was recorded using low compliance capillary water per fusion manometric system. EM was administered intravenously during phase I and after meal, and blood samples were drawn for measuring plasma motilin concentra- tions. RESULTS ①Plasma motilin levels showed cyclical fluctuations in different phases of MMC, and plasma motilin reached peak during phaseⅢ and lowest during phase Ｉ. ②EM induced phase Ⅲ -like contractions in the antrum and duodenum, which was not accompanied by a peak in plasma motilin level. The optimum dose of EM for resulting in a premature phaseⅢ with the same characteristics as the spontaneously occurring phaseⅢ was established to be 0. 5 mg.kg-1. The dose of 10 mg.kg-1 EM induced gas- trointestinal continuous contractions and duodeno-gas-tric retrograde peristalsis which was associated with retching and vomiting. ③Atropine obviously inhibited EM-induced phase Ⅲ activity in the antrum and duodenum. GEM powerfully enhanced postprandial gastrointestinal contractile activity. CONCLUSIONS The results suggests that EM is a potent prokinetic agent. The mechanism is not related to the release of motilin, but may be mediated partially by cholinergic pathway.

This study investigated the tight junction (TJ) protein expression of the intestinal mucosa in a rat tail-suspension model under simulated weightlessness. Twenty-four Wistar rats were randomly divided into three groups: CON group (n=8), control; SUS-14 d group (n=8), tail-suspension for 14 days; SUS-21 d group (n=8), tail-suspension for 21 days. Occludin and Zonula Occluden-1 (ZO-1) expression levels were determined by immunohistochemical analysis and mRNA fluorescent quantitative PCR. Plasma levels of diamine oxidase (DAO) and d-lactate were determined using enzymatic spectrophotometry. Immunohistochemical results for occludin and ZO-1 showed disruption of the TJs in the intestinal mucosa in SUS-14 d and SUS-21 d groups. The expression levels of occludin and ZO-1 in SUS-21 d group were lower than those in SUS-14 d group, and significantly lower than those in CON group (Occldin: 0.86±0.02 vs 1.01±0.03 vs 1.63±0.03 and ZO-1: 0.82±0.01 vs 1.00±0.02 vs 1.55±0.01, P<0.01). Moreover, the levels of plasma DAO and d-lactate in SUS-21 d group were higher than those in SUS-14 d group, and significantly higher than those in CON group (DAO: 27.58±0.49 vs 20.74±0.49 vs 12.94±0.21 and d-lactate: 37.86±0.74 vs 28.26±1.01 vs 17.76±0.91, P<0.01). There were significant negative correlations between occludin or ZO-1 expression levels and DAO (r (2)=0.9014, r (2)=0.9355, P<0.01) or d-lactate levels (r (2)=0.8989, r (2)=0.9331, P<0.01). Occludin and Zo-1 were reduced in intestinal mucosa both in mRNA and protein levels in the rat tail-suspension model. The significant negative correlations between expression levels of TJs and plasma levels of DAO or d-lactate support the hypothesis that intestinal permeability is increased due to a decrease in TJ protein expression during tail-suspension from 14 days to 21 days.

AIM　To observe the effect of intravenous injection of erythromycin (EM) on interdigestive migrating motor complex (MMC) and postprandial gastrointestinal contraction in conscious dogs , and to study its possible mechanism. METHODS　Gastrointestinal contractile activity was recorded using low compliance capillary water perfusion manometric system. EM was administered intravenously during phaseⅠ and after meal, and blood samples were drawn for measuring plasma motilin concentrations. RESULTS　①Plasma motilin levels showed cyclical fluctuations in different phases of MMC, and plasma motilin reached peak during phaseⅢ and lowest during phaseⅠ.②EM induced phaseⅢ-like contractions in the antrum and duodenum, which was not accompanied by a peak in plasma motilin level. The optimum dose of EM for resulting in a premature phaseⅢ with the same characteristics as the spontaneously occurring phaseⅢ was established to be 0.5 mg*kg-1. The dose of 10 mg*kg-1 EM induced gastrointestinal continuous contractions and duodeno-gastric retrograde peristalsis which was associated with retching and vomiting. ③Atropine obviously inhibited EM-induced phaseⅢ activity in the antrum and duodenum. ④EM powerfully enhanced postprandial gastrointestinal contractile activity. CONCLUSIONS The results suggests that EM is a potent prokinetic agent. The mechanism is not related to the release of motilin, but may be mediated partially by cholinergic pathway.

Objective To observe the effect of sevoflurane and target controlled infusion (TCI)propofol on the oculocardiac reflex (OCR) in patients with paediatrie strabismus surgery.Methods One hundred and thirty-eight patients ASA Ⅰ-Ⅱ undergoing strabismus surgery were randomly allocated to six greups(propofol groups:P_1,P_2,P_3 group; sevoflurane groups:S_1,S_2,S_3 group,23 cases for each) according to target bispectral index (BIS) of 60,50 and 40.In propofol groups continuous infusion of propofolremifentanil [0.2μg/(kg·min)] was adjusted towards target BIS value.The sevoflurane-remffentanil [0.2μg/(kg·min) ] concentration with 50% N2O/O2 was adjusted toward target BIS too.The incidence of OCR and the lowest heart rate(HR) and BIS were recorded.Results The incidence of OCR were 73.9%(17/23),39.1%(9/23),17.4%(4/23),56.5%(13/23),26.1%(6/23),8.7%(2/23)in P_1,P_2,P_3,S_1,S_2,S_3 soup,P_2 and P_3 group were lower than P_1 group,S_2 and S_3 soup were lower than S_1 group,P_3,S_3,S_2,P_2 group were lower than S_1 and P_1 soup,S_1 soup was lower than P_1 group,S_2 soup was lower than P_2 group,S_3 group was lower than P_3 group,there was significant difference(P < 0.05 ).The densities of TCI propofol in P_2 and P_3 soup were higher than that in P_1 group [ (4.3 ±0.2),(5.5±0.1 ) mg/L vs (3.2±0.1 ) mg/L ] (P <0.05),and the densities of sevoflurane in S_2 and S_3 group were higher than that in S_1 group [ ( 1.8±0.3 )%,(2.3±0.2 )% vs(1.3±0.2 )% ] (P<0.05 ).The end-tidal concentration was different with difference of BIS too.Conclusions OCR is relevant to the depth of anesthesia.BIS values of 40-50 seem adequate for the inhibition of OCR.The results suggest that BIS may be a valuable tool during propofol-remifentanil or sevoflurane-remifentanil anesthesia for strabismus surgery in children.The incidence of OCR is higher in propofol than in sevoflurane at the same BIS.

OBJECTIVETo correlate the clinical characteristics with mutations of the STK11 and FHIT genes in 16 patients with Peutz-Jeghers syndrome (PJS).

METHODSPotential mutations in the coding regions and flanking sequences of the STK11 and FHIT genes were detected with PCR and Sanger sequencing.

RESULTSOf the 16 patients with PJS, 8 had novel mutations in the coding region of the STK11 gene, 1 had a previously reported mutation. 1 carried a mutation in the exon 10 of the FHIT gene, which is a non-coding region. None of the mutations was detected in the immediate family members. None of the patients with STK11 gene mutations had mutation in the FHIT gene. The mutation rate of the STK11 gene among patients with PJS was 56.25%.

CONCLUSIONMutations of the STK11 gene are the major cause of PJS. Few such patients had mutations of the FHIT gene. Mutations of the FHIT gene may play a part in the pathogenesis of PJS.

Acid Anhydride Hydrolases ; genetics ; Adolescent ; Adult ; Base Sequence ; Child ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Neoplasm Proteins ; genetics ; Pedigree ; Peutz-Jeghers Syndrome ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Young Adult

This study investigated the tight junction (TJ) protein expression of the intestinal mucosa in a rat tail-suspension model under simulated weightlessness.Twenty-four Wistar rats were randomly divided into three groups:CON group (n=8),control; SUS-14 d group (n=8),tail-suspension for 14 days;SUS-21 d group (n=8),tail-suspension for 21 days.Occludin and Zonula Occluden-1 (ZO-1) expression levels were determined by immunohistochemical analysis and mRNA fluorescent quantitative PCR.Plasma levels of diamine oxidase (DAO) and d-lactate were determined using enzymatic spectrophotometry.Immunohistochemical results for occludin and ZO-1 showed disruption of the TJs in the intestinal mucosa in SUS-14 d and SUS-21 d groups.The expression levels of occludin and ZO-1 in SUS-21 d group were lower than those in SUS-14 d group,and significantly lower than those in CON group (Occldin:0.86±0.02 vs 1.01±0.03 vs 1.63±0.03 and ZO-1:0.82±0.01 vs 1.00±0.02 vs 1.55±0.01,P＜0.01).Moreover,the levels of plasma DAO and d-lactate in SUS-21 d group were higher than those in SUS-14 d group,and significantly higher than those in CON group (DAO:27.58±0.49 vs 20.74±0.49 vs 12.94±0.21 and d-lactate:37.86±0.74 vs 28.26±1.01 vs 17.76±0.91,P＜0.01).There were significant negative co rrelations between occludin or ZO-1 expression levels and DAO (r2=0.9014,r2=0.9355,P＜0.01) or d-lactate levels (r2=0.8989,r2=0.9331,P＜0.01).Occludin and Zo-1 were reduced in intestinal mucosa both in mRNA and protein levels in the rat tail-suspension model.The significant negative correlations between expression levels of TJs and plasma levels of DAO or d-lactate support the hypothesis that intestinal permeability is increased due to a decrease in TJ protein expression during tail-suspension from 14 days to 21 days.