The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification.Here, the mutant (CM113R)7 αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd2+ ions.Different reaction pathways, intermediates, and products could be recognized.Cd2+-GSH reaction was highly dependent on the solution pH.The Cd(GSH)2 was formed at pH 7.4, while the Cd(GSH)2 and Cd2(GSH)2 were formed at pH 9.0.Cd2(GSH)2 was formed by two possible pathways: (1) A Cd2+ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)2, and then the second Cd2+ ion quickly incorporated with the deprotonated amino group of Cd(GSH)2 to form Cd2(GSH)2;(2) Two Cd2+ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd2(GSH)1, and the second GSH molecule quickly bounded to Cd2+ ions to form Cd2(GSH)2.This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.

ObjectiveTo evaluate the reliability and validity of the of Disability Assessment Schedule Ⅱ of World Health Organization(WHO-DASⅡ) applied in assessment of the function of disabled athletes.Methods56 disabled people,26 wheelchair basketball athletes and 30 patients,were sampled and completed WHO-DASⅡ questionnaire.The test and re-test had been administrated in 10 days.Barthel Index(BI) and Sports Classification Test had been implemented for the functioning assessment.ResultsThe test-retest reliability for WHO-DASⅡ in six dimensions were: 0.978,0.807,0.938,0.877,0.998,0.935,0.986 and all were in very significant level(P<0.01).There were negative correlation between WHO-DASⅡ scores and BI in total and in every dimension,the correlation coefficients were:-0.77,-0.17,-0.71,-0.82,-0.33,-0.73,-0.61.The correlation coefficients between WHO-DASⅡ total and six dimensional scores and the grade of sports classification were:-0.741,-0.378,-0.806,(-0.541),-0.293,-0.510,-0.677.ConclusionWHO-DASⅡ can be used in assessing the function of disabled athletes.

Hela is the cell line of adenocarcinoma of the uterine cervix, and human papillomavirus (HPV) 18 shows positive. We delivered siRNA with target specifically to HPV18 E7 mRNA into nude mice Hela tumor xenografts by nanopatch to inhibit the HPV gene expression, and further to study the superiority, the best action time and concentration of siRNA of using nanopatch to transfer siRNA in vivo. We designed siRNA that target specifically to HPV18 E7 mRNA (siE7) and checked the effect of siE7 in vitro. Tumor xenografts were transfected with siE7 and GenEscort III by nanopatch. Expression of HPV18 E7 mRNA and protein were detected 0 hours, 24 hours, 48 hours, 72 hours after transfection with PT-PCR and Western blot, and the best action time was analyzed using nanopatch to thansfect siRNA in vivo. We transfected GenEscort III and siE7 of Different concentration into tumor xenografts respectively by nanopatch and intraperitoneal injection. Expression of HPV18 E7 mRNA and protein was detected 72 hours after transfection by PT-PCR and Western blot, to analyze the best action concentration of siRNA and the superiority of using nanopatch to thansfect siRNA in vivo. The results proved that SiE7was efficient to inhibit expression of HPV18 E7 mRNA and to advance Hela apoptosis in vitro. SiE7 transfected by nanopatch into xenografts could inhibit effectively expression of HPV18 E7 mRNA and protein. The best action time and concentration of siRNA of using nanopatch to thansfect siRNA in vivo are 72 hour post-transfection and 2 micromol/L siE7. To compare intraperitoneal injection in delivering siRNA in vivo, the effect of nanopatch is very predominant. It can be well concluded that Nanopatch can effectively transfer siRNA in vivo, which can effectively inhibit the HPV gene expression.
Animals ; Apoptosis ; DNA-Binding Proteins ; genetics ; Female ; Gene Expression Regulation, Viral ; HeLa Cells ; Humans ; Mice ; Mice, Nude ; Nanostructures ; administration & dosage ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; RNA, Small Interfering ; administration & dosage ; Transfection ; Uterine Cervical Neoplasms ; therapy ; Xenograft Model Antitumor Assays

Objective To explore the higher dangerous factors,the early clinical performances and its contents of neonatal pulmonary hemorrhage(NPH).Methods The clinical performances,chest radiograms and autoptical pathological materials of 66 cases of newborns who died of NPH at our neonatal department during 1993 to 2003 were reviewed and analyzed.Results The higher dangerous factors of NPH were premature delivery/low birth weight,serious diseases lead to hypoxia and severe infections.The early clinical performances of NPH were the suddenly aggravation of dyspnea and the increasing of moist sounds.The early X-ray performances were lower penetrance of lung fields extensively and well-distributly with path clouds,the intercostals space usually increased.According to the autoptical(patho)-logy,this X-ray perfomance indicated the edema of the pulmonary with small amount of hemorrhage.Conclusion The patients with the higher dangerous factors and the early clinical performances of NPH,must be diagnosed and interfered it as early as possible to reduce the mortality of NPH.

Purpose To explore the relationship between the mutations of epidermal growth factor receptor ( EGFR) and KRAS genes and clinicopathological characteristics in patients with non-small cell lung cancers (NSCLC). Methods Clinical samples from 431 NSCLC patients were obtained for EGFR and KRAS gene analysis. PCR based direct DNA sequencing was used to investigate mutations in exon 18-21 of EGFR gene and codon 12 and 13 of exon 2 of KRAS gene. Results The overall EGFR mutation rate of primary NSCLC was 53. 6% (231/431) in this study cohort and eight cases showed double EGFR mutations. Mutation rates in female and male were 65. 2% (122/187) and 46. 9% (98/209), respectively. The mutation rate was higher in patients with non-smokers and adeno-carcinoma and adenosquamous carcinoma subtypes than in their counterparts (P<0. 05), with the percentage of 57. 2% (124/216), 60. 3% (199/330), 42. 9% (6/14), respectively. In squamous cell carcinomas and other subtypes, EGFR mutation rates were 11. 6% (5/43) and 11. 1% (1/9), respectively. The EGFR mutation types included exon 18 point mutations (4. 0%, 9/227), exon 19 deletion mutations (4. 5%, 101/227), exon 20 insert or point mutations (9. 7%, 22/227) and exon 21 point mutations (41. 4%, 94/227). Activating mutations of KRAS gene were detected in 7. 8%(31/396) of NSCLC. Twenty-eight patients showed codon 12 mutations ( G>T, G>A, G>C) , and three patients had codon 13 mutations ( G>A, G>T) . Most of these mutations were G to T transversion (64. 5%, 20/31). Conclusion Polymerase chain reaction-direct sequencing is a reliable and effective method for the detection of the EGFR and KRAS gene mutation in NSCLC patients. The mutation rate of EGFR is higher in Chinese patients, especial-ly in non-smoking female patients with adenocarcinoma.

Brain Concussion ; complications ; Hearing ; Hearing Disorders ; prevention & control ; Humans ; Thioctic Acid ; therapeutic use

AIM:To establish a HPLC fingerprint of Stellera chamaejasme L. from Inner Mongolia Region. METHODS: The RP-HPLC method was used with Akzonobel Kromasil C_ 18 (250 mm?4.6 mm, 5 ?m) the acetonitrile-0.5% phosphoric acid (gradient elution) was used as mobile phase, analytic time was 60 min, and detective wavelength was at 297 nm, the column temperature of 15℃ were adopted. RESULTS: The HPLC fingerprint of Stellera chamaejasme L. set up showed that 14 peaks were co-possessing in different sources. The results of method validation met technical standard of fingerprint, the similarities of Stellera chamaejasme L. were 0.9 to 1.0. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of Stellera chamaejasme L. from Inner Mongolia Region.

Objective to evaluate the impacts of 6 standard comparible tables of biparietal diameter(BPD)-gestational age to the second trimester Down syndrome screening and the individual risk rate for pregnant woman. Methods A total of 25 346 pregnant woman with gestational age be-tween 12 to 20 weeks were recruited and analyzed in the study,including 32 Down syndrome. For each pregnant woman,we calculated the gestation-al age by six different BPD-gestational age tables,and the prenatal screening risk value of gestational age. then ROC curve,detection rate and posi-tive rate were used to evaluate the effect of different control tables on the screening results. In addition,individual risk rate was used to determine the influence of gestational age deviation on pregnant woman. Results According to the ROC,we found that our self-designed BPD table had the best AUC(0.972),while the table designed by a hospital in Shanghai had the least AUC(0.923). the difference was significant by statistic test(P =0.045). With the same detection rate of 75%,we found our self-designed table had the least positive rate with only 2.3%,and the cutoff was 1∶280, while the table of Chinese Obstetrics and Gynecology had the highest ratio(4.8%),and the cutoff was 1∶345. Our results suggest that once gesta-tional age is estimated to be higher than actual age,the risk will be higher and the positive rate will increase. However,if gestational age is estimated to be smaller,the risk will be smaller and the negative rate will increase. Conclusion the impacts of different tables on Down syndrome screening are different. Our self-designed BPD table is the most effective to estimate gestational age,and the control table of Hongkong is ranked No.2. the wrong estimation of gestational age will largely affect the individual risk of pregnant woman.

Objective To study a feasible technology plan for information communication between doctors and patients.Methods Based on three-layer B/W/S and Windows2003 Server,MS SQL Server was used for database administration.The Apache Tomcat 5.5 was adopted as Web server and Web services were realized by using JSP(Java Server Page).Results Health care information was collected and integrated in medical information platform of clinical department which was connected into Internet directly.The information platform established communication way between doctors and patients and it had functions for chief complaint,diagnosis,reservation and revisit.Conclusion Communication between doctors and patients are realized.

Based on target cycling amplification and 3 ,4 ,9 ,10-perylenetetracarboxylic dianhydride derivative functionalized singal probe, an ultrasensitive electrochemiluminescence ( ECL) sensor was designed for the detection of lead ions. The hairpin substrate DNA was immobilized on the electrode through molecular self-assembly. In the presence of Pb2+and DNAzyme, the substrate was cleaved with single strand DNA fragments left on the electrode surface. Meanwhile, the target and DNAzyme was released for another cleaving circularly. As a result, the single strand DNA fragments hybridized with the assist hairpin probe H1, which leaded to the fabrication of H2 labeled with the 3 , 4 , 9 , 10-perylenetetracarboxylic dianhydride derivative functionalized hollow gold nanoparticles. With the increasing concentration of Pb2+, much more signal probe was been captured and the ECL signal of the biosensor in peroxydisulfate ( S2 O2-8 ) solution would increase. An ECL assay demonstrates that the sensor has a good linear response to Pb2+ concentration in the range of 1í10-12 mol/L-1í10-6 mol/L, with a detection limit of 1í10-12 mol/L. The fabricated sensor shows good selectivity toward Pb2+against other common metal ions.