Objective To verify the detection performance of glycated hemoglobin (HbA1c)by using the BIO-RAD D-10 high performance liquid chromatography method,and to evaluate whether the detection performance can satisfy the requirements of the laboratory and clinical diagnosis and treatment.Methods The method of CLSI EP1 5A2 was used to verify the accuracy and preci-sion.The linear correlation and reference range were verified respectively by linear regression analysis and the method of the WS/T 402 2012.Results The calibration value of HbA1c calibrator level 1 was 5.3% and the validation limit of measured results was 1. 63%-8.78%.The calibration value of HbA1c calibrator Level 2 was 10.0%,and the validation limit of measured results was 4.76%-13.64%.Both calibration values were included in the 95% confidence range of measured values,so the accuracy verifica-tion passed.Laboratory variation coefficient (CV)for HbA1c with concentration of 5.24% was 1.01% and for 9.84% was 0. 54%,both less than the precision that the manufacturer declared,so the precision verification passed.The equation of linear regres-sion analysis for HbA1c linearity calibrator measured value and target value wasY =1.008X +0.023,with correlation coefficient in-dex r 2 of 0.999,so the linearity was good.The measured results of 20 healthy adults specimens were in the reference interval(4.1%-6.2%)that the manufacturer provided.Conclusion Accuracy,precision,linear correlation and reference range of HbA1c meas-ured by BIO-RAD D-10 HPLC detection method were verified.It can provide the accurate and credible measurement results for clin-ical diagnosis and treatment,and can be used in the detection of clinical specimens.

Objective To explore the effect of dendrobium nobile polysaccharides on the expression of the WT 1 gene in my-eloid leukemia cells.Methods The CCK8 assay was used to detect the half maximal inhibitory concentration(IC50 )of dendrobium nobile polysaccharides in 3 kinds of leukemia cells;the each kind of leukemia cells were divided into the treatment group and the control group.The cells in the control group maintained the normal growth,while which in the treatment group were given the den-drobium nobile polysaccharides stimulation.The Hoechst33258 staining was used to detect the apoptosis situation of the cells in the two groups.The WT l gene expression level was detected by the real-time PCR and the protein expression levels of WT1,53 and BAX were detect the Western blot.Results Dendrobium nobile polysaccharides had the similar IC50 values in 3 kinds of myeloid leukemia cells,which were (110.71±6.49),(104±48.50),(96.66±5.10)mg/mL respectively,the difference among them had no statistical significance (P >0.05);the apoptosis rate of the treatment group was higher than that of the control group (P <0.05);the expression levels of WT1 gene and protein in the treatment group were decreased compared with the control group(P <0.05), while the expression of P53 and BAX protein was increased.Conclusion Dendrobium nobile polysaccharides can obviously decrease the expression level of WT1 protein,and has a certain killing effect on myeloid leukemia cells.

Objective The present study analyzed the performances of hearing impaired children and normal hearing children at age 3 on the open set monosyllables test to provide reference for rehabilitation .Methods 30 typi-cal children and hearing impaired children at age 3 were randomly selected and tested using the open -set monosylla-ble test in evaluation of auditory response to speech (EARS) .Results ① There were significantly differences be-tween different test items in both groups ,the decreasing order of the score was tone> vowel>consonant>word(P<0 .01) ,and the scores on vowels ,consonants and words of hearing -impaired children were respectively signifi-cantly lower than that in normal hearing children(P<0 .001) .②There were similarities and differences between the mistakes of typical children and hearing impaired children .Blade-palatals and laterals in both groups achieved the lowest accuracy .While the accuracies on bilabials ,labiodentals ,nasals and plosives were highest in typical children , the accuracy on velar was highest in hearing impaired children .And the accuracies of consonants and nasal vowels on each place and manner of articulation in hearing -impaired children were significantly lower .Conclusion The study showed that the performances on vowels ,consomants and words of hearing -impaired children aged 3 in the open set monosyllables test were poorer than that in normal hearing children .The main errors were between the same places or methods of articulation ,especially for Blade -palatals and laterals .It is possible related to the deficiency of lan-guage input with the hearing impairment and the age with HA/CI .

Objective To investigate the changes of IL-8 in peripheral blood of patients with pustular psoriasis after acitretin treatment.Methods Dual antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to measure the IL-8 levels in the peripheral blood of 30 patients with pustular psoriasis and 30 patients with psoriasis vulgaris before and after treatment,and in 30 normal human controls.The relationship between the IL-8 level and disease severity was assessed for patients with pustular psoriasis.Results After acitretin treatment,the condition was improved greatly in patients with pustular psoriasis,together with a significant decrease in the level of IL-8 in the peripheral blood(57.07 ± 12.02 pg/ml vs.96.84 ± 14.68 pg/ml,P ＜ 0.05).Conclusion IL-8 may be involved in the therapeutic mechanism of acitretin in pustular psoriasis.

People wearing positive pressure biological protective suits might face some potential safety risks when work -ing in a high-risk biological contamination environment .These risks could be effectively prevented by simulation experi-ments.This paper elaborates on the progress in risk studys on positive pressure biological protective suits in terms of dynam -ic detection methods and risk assessment , and points out the problems to be solved and the direction of future development .

Objective To build an antibody microarray based on direct labeling strategy for microalbnrninuria measurement, and evaluate it's technical potentiality for clinical application. Methods Urine samples of diabetic patients were collected. Antibody microarrays were constructed by preparation of array support, array fabrication, then protein assay and data analysis were performed. Procedure conditions for each step especially the labeling of samples were optimized. The set-ups were evaluated in terms of sensitivity, specificity and reproducibility. Urinary albumin excretion in the samples was detected by fabricated protein array, which was compared to that detected with immunoturbidimetry. Results The signal intensity was best when protein quality ratio of pure albumin or urine sample against NHS-biotin was 2:1. A calibration curve with a correlation coefficient of 0.9995 was established. The lower limit of detection was 0.0617 mg/L. Interehip and intrachip variation studies conducted on patient urine demonstrated CVs as 6.78%-9.22% and 3.35%-7.59%, respectively. Compared with the immunoturbidimetry, the antibody microarray was able to detect the extremely lower grade albumin in urine samples. The correlation coefficient of the results obtained by the two methods was 0.9199 (P <0.01). Conclusion An antibody microarray based on direct labeling strategy for microalbuminuria measurement is successfully established, which is comparable to immunoturbidimctry in its accuracy and will have great potential for clinical use with its high throughput, sensitivity, specifity and reproducibility.

Objective To investigate the effects of two cholesterol-lowering probiotics, DM9054 (Lac-tobacillus Rhamnosus GG, LGG) in combination with 86066 (Lactobacillus plantarum WCFS1, LP), on the metabolism of bile acid via a rat model of non-alcoholic fatty liver disease (NAFLD) and the possible mecha-nism. Methods Twenty-one SD male rats were randomly divided into three groups including control group, NAFLD model group and probiotics intervention group. Rats in the control group received normal diet. The rat model of NAFLD was established by feeding rats with chronic high fat diet (45% of calories derived from fat di-et) for 20 weeks. Rats in the probiotics intervention group were given high fat diet together with cholesterol-low-ering probiotics through oral gavage. General indexes of each group including body weight and the levels of tri-glyceride, cholesterol and CK18-M30 in serums samples were detected. The expression of cholesterol 7-alpha hydroxy-lase (CYP7A1), fibroblast growth factor receptor 4 (FGFR4), farnesoid X receptor (FXR), fibroblast grwoth factor 15 (FGF15) and apical sodium-dependent bile acid transparter(ASBT) at mRNA level were de-tected by using real-time polymerase chain reaction (real-time PCR). Western blot assay was used to detect the protein expression of CYP7A1, FXR in liver tissues and ASBT in ileum tissues. The expression of FXR in liver and ileum tissues were analyzed by immunohistochemistry. Results Rats with NAFLD showed loss of body weight and decreased levels of the serological markers after treating with the probiotics (P<0. 05). Compared with the rats in model group, enhanced expression of CYP7A1 and inhibited expression of FXR in liver tissues, activated FXR-FGF15 pathway in ileum tissues as well as down-regulated expression of ASBT in ileum tissues were detected in rats receiving probiotics intervention (P<0. 05). No significant difference in the expression of FGFR4 at mRNA level was observed between NAFLD rats with or without probiotics intervention (P>0. 05). Conclusion Probiotics intervention might up-regulate the expression of CYP7A1 by suppressing the FXR path-way in liver tissues and inhibiting the expression of ASBT in ileum tissues. Treating NAFLD rats with cholester-ol-lowering probiotics could activate the FXR-FGF15 pathway in ileum tissues and enhance the metabolism of bile acid, which contributed to the alleviation of NAFLD.

Objective To investigate the effect of shenfu injection on immune function in severe trauma patients.Methods 60 severe trauma patients were divided into the control group (n =30)and shefu group (n =30) by random number table.Other 30 cases were chosen as the health control group at the same phase.All patients were received conventional treatments,however,patients of the shenfu group were additionally received the shenfu injection treatment in the early stage.The CD +3 ,CD +4 ,CD +8 cell,human leukocyte antigen (HLA -DR),interleukin -1(IL -1),interleukin -6(IL -6)were detected on 3rd and 7th day by double -antibody sandwich enzyme -linked immu-nosorbent assay (ELISA).Results Compared to the health control group,the IL -1,IL -6 in the control and shenfu group were significantly higher than the health control group(f=7.128,q =9.212,10.112,all P <0.05).The IL -1and IL -6 in the control and shenfu group were significantly increased on 3rd day (t =11.126,10.013,all P <0.05)and decreased on 7th day(t =17.121,14.213,all P <0.05).The IL -1 and IL -6 in shenfu group were sig-nificantly lower than that of the control group(χ2 =4.113,10.117,all P <0.05).The CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were significantly lower than the health control group(f=11.071, q =10.229,12.032,all P <0.05).On 3rd day,the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly increased(t =10.013,P <0.05).On 7th day,CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were both increased(t =11.126,15.932,all P <0.05).And the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly higher than the control group(χ2 =3.771,P <0.05).Conclusion Shenfu injection can regulate immune function in severe trauma and improve clinical treatment.

Objective To study the effect of oxymatrine (OMT) combined with radiotherapy on the growth of HO8910 cells of human ovarian cancer in vitro.Methods All human ovarian cancer samples were divided into four groups:control group,treated with culture solution; OMT group,treated with 4 mg/ml OMT; radiotherapy group,treated by 4 Gy radiotherapy; and combination group,firstly treated by 4Gy radiotherapy,and then cultured with 4 mg/ml OMT.Observed the changes of cell morphology by invert microscope at 10,24,48,and 72 hours respectively.Flow cytometry was adopted to detect cell apoptosis.Results ① invert microscope observation showed that compared with the control group,HO8910 cells demonstrated apoptosis of diminution in volume,thickening in cytoplasm,and gathering in nucleus in all the other three groups at 24 and 48 hours.② Flow cytometry showed that apoptosis rate of HO8910 cells and cell population in G1 phase increased in the combination group,which were significantly higher than the other three groups (P＜0.05).Conclusion Both OMT (4 mg/ml)and radiation (4Gy)can induce cell apoptosis,while the combination of them showed better therapeutic results.

Objective To screen the variation in NeuroD1 gene and to study its function in vitro and its clinical phenotypes and genetic characteristics in Chinese early-onset type 2 diabetic probands. Methods PCR-direct sequencing of NeuroD1 gene was performed in 85 early-onset type 2 diabetic probands, 95 late-onset type 2 diabetics with strong diabetic history and 87 non-diabetic control subjects. Distributions of the identified variation were calculated and compared among the three groups. Expression vectors with mouse NeuroD1 (mND1)cDNA wild type or mutant type and reporter vectors with human insulin promotor-linked luciferase were constructed. Then the above vectors were co-transfected into rat INS-1 cells. Relative luciferase activities were measured to compare transcriptional activities of insulin gene between WT and MT. Results S159P (T→C), a new mutation was identified in a proband, which was co-segregated with diabetes in 4 carriers from the paternal side. The functional study showed that the S159P mutant exhibited a 25% reduction in transcriptional activity of insulin gene as compared with the wild type. A45T (G→A), a common variation was identified. The AA + GA genotypic frequencies were markedly increased in early-onset type 2 diabetic probands as compared with late-onset type 2 diabetic probands and non-diabetic control subjects (P=0.006 and P=0.014, respectively). Conclusion The novel S159P mutation in the NeuroDl gene seems to contribute to the development of diabetes in the Chinese early-onset type 2 diabetic family. The A45T variation may increase susceptibility to or be in disequilibrium with early-onset type 2 diabetes mellitus in Chinese population. In addition, the A45T variation may affect the onset pattern of type 2 diabetes mellitns, such as early-onset but not late-onset type 2 diabetes mellitus.