Objective To evaluate the effect of various sampling errors on ELISA test results. Methods Standard sample volume,standard sample volume reducing 1,2,3,4 μL or adding 1, 2,3 μL were respectively pipetted into the wells of a microplate,follwed by routine operation of ELISA test. Then the influence of various sampling errors was analyzed on ELISA test results of HBsAg, HCV and TP. Results S/CO value was increased with the increase of sample volume. The statistical difference of mean S/CO value of HBsAg and TP was only found between sample volume adding 3 μL group and control group(P<0.05). For HCV result, there were significant differences between standard sample volume adding 2,3μL or reducing 3,4μL groups and control group(P<0.05), while no obvious differences were found in the other groups(P>0.05). The difference of mean positive rate between ex-perimental groups and control group showed an increasing tendency with the reduction of sample vol-ume,and significant differences in HBsAg, HCV and TP results were also found between sample vol-ume increase groups and reduction groups(P<0.05). Conclusion Various sampling errors influence ELISA test results to different degrees,and the extent increases with the reduction of standard sample volume.

Adult ; Device Removal ; methods ; Ductus Arteriosus, Patent ; Heart Septal Defects, Atrial ; therapy ; Humans ; Male

Objective To study the diagnostic value of fractional exhaled nitric oxide(FeNO)in non-typical bronchial asthma.Methods Ninety-five patients with unknown-cause respiratory symptoms including wheezing,cough and breathlessness were enrolled.FeNO was measured by nitric oxide analyzer.The clinical symptoms and bronchial bronchodilator test were defined as golden standard for asthma diagnosis.The diagnostic value of FeNO was assessed and the optimal operating point of FeNO test was determined by means of the receiver operating characteristic(ROC)curve.Results Among 95 patients,44 cases were diagnosed as asthma,51 cases were diagnosed as non-asthma.The level of FeNO of asthma patients were higher than that of non-asthma patients[(55.2±14.0)nmol/L vs.(18.9±5.2)nmol/L,P＜0.01].A non-linear correlation of FeNO with FEV1％ was revealed in the cases with asthma(r=-0.162,P＞0.05).Area under ROC curve was 0.858.The optimal diagnostic cut off point was 36 nmol/L which was capable of differentiating asthma and non-asthma with sensitivity of 80.2％,specificity of 79.5％,positive predictive value of 85.4％,negative predictive value of 83.3％ and accuracy of 85.9％.Conclusions FeNO test may be helpful in the diagnosis of non-typical asthma with high sensitivity and specificity,which can also improve the diagnostic effectiveness and avoid misdiagnosis,missed diagnosis when combined with lung function test.

Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

Objective To investigate the influence of total progressively motile sperm count(TPMSC) after treatment on clinical outcomes of intrauterine insemination(IUI) with the husband′s sperm in ovulation-promoting cycles.Methods The clinical data in 4179 cases undergoing IUI with the husband′s sperm in ovulation-promoting cycles were retrospectively analyzed.The correlation between clinical pregnancy rate and TPMSC was analyzed.Results Among all the clinical data,TPMSC was to 100×106 in occasional live sperm.TPMSC<0.15×106 was in 15 cases,1 case had pregnancy (live sperm was occasionally seen on IUI day after sperm processing).Ten cases of TPMSC >60×106 had no pregnancy.A total of 4 154 cases of TPMSC (0.15-60.00)×106 were analyzed.The female age,duration of infertility,number of follicles and endometrial thickness(EDM) had no statistical differences among various groups.The clinical pregnancy rate was 13.5%(576/4 154),the group with the highest clinical pregnancy rate was (5.00-<10.00)×106.But there was no statistically significant difference in clinical pregnancy rate among groups(P=0.133).Conclusion Performing IUI in PMSC (0.15-60.00)×106 after processing can get preferable pregnancy rates.

Objective To investigate the effect of dexmedetomidine on the intubation reaction during intratracheal intubation in the patients in the emergency department .Methods Fifty adult patients needing emergent tracheal intubation in the emergency de-partment of this hospital were divided into the dexmedetomidine group and the middazolam group ,25 cases in each group .The dexmedetomidine group was given dexmedetomidine(1 μg/kg) before tracheal intubation and the midazolam group was given mid-azolam(0 .1 mg/kg) before tracheal intubation .Results BP and HR during tracheal intubation ,at 1 ,3 min after tracheal intubation in the dexmedetomidine group were significantly decreased compared with the midazolam group (P<0 .05);SpO2 at 1 min before tracheal intubation and during tracheal intubation in the dexmedetomidine group had no statistical difference compared with before drug administration(P>0 .05);SpO2 at 1 min before tracheal intubation and during tracheal intubation in the middazolam group was decreased ,which showed the statistical difference compared with before drug administration (P<0 .05) .Conclusion Dexme-detomidine is more effective than midazolam in alleviating cardiovascular responses during intratracheal intubation ,moreover has no influence on the patient′s respiratory function .

Animal models with kidney disease are generally divided into two types. One belongs to the models which imitate human kidney disease by the artificial operations, such as anti-glomerular basement membrane antibody nephritis, Heymann nephritis, anti-Thyl. 1 antibody nephritis, BSA nephritis and puromycin nephropathy. The other one pertains to the models which make themselves kidney disease, and appear the pathological characteristics naturally as like as human, such as HIGA mice with IgA nephropathy and NZB/WF1 and MRL/1pr mice with lupus nephritis. In addition,the transgenic animal models with kidney disease can also be established by the modern molecular biologic techniques including gene knockout and siRNA transfection. As for the studies related with kidney disease in pharmacodynamics and pharmacology of Chinese herbal medicine (CHM), it is important to understand deeply the features of each animal model with kidney disease, and select accurately the proper models according to the different experimental objectives, and then, build the special models provided with the combination of disease with syndrome in traditional Chinese medicine (TCM). Therefore,it is the developmental direction for the further study to establish animal models with kidney disease, which should possess the characteristics of syndrome in TCM.
Animals ; Diabetic Nephropathies ; etiology ; Disease Models, Animal ; Humans ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Streptozocin

Aged ; Female ; Fibroma ; pathology ; Fibroma, Ossifying ; pathology ; Gingival Neoplasms ; pathology ; Humans

In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.