Objective To investigate the role of vascular endothelial cell growth factor (VEGF), nm23 and microvessel density (MVD) in the pathogenesis of malignant melanoma(MM). Methods The specimens from forty-seven cases of malignant melanoma and 10 cases of melanocytic nevus were examined for VEGF, nm23 and CD34 by immunohistochemistry. The density of microvessels was calculated. Results Positive VEGF expression rate and MVD in malignant melanoma were significantly higher than those in melanocytic nevus(both P

Objective To investigate the relationship of vitamin D with the intestinal development and study the expression of vitamin D receptor (VDR) in the duodenum of C57BL/6 mice at different developmental stages.Methods Quantitative PCR (qPCR),histology using H&E staining and immunofluorescence staining,and Western blotting (WB) were performed to elucidate the expression of VDR in mice intestine at different growth and developmental stages.Results The peak of VDR mRNA expression reached on 21 d.The pathological result showed that VDR mainly distributed in the cytoplasm of epithelial cells and smooth muscle cells in the mouse duodenum.WB result indicated that there was no nuclear translocation of VDR protein in the mouse duodenum.Conclusions This study demonstrates the regularity of expression of VDR in the mouse duodenum during its development,and contributes to understanding the function of VDR in the intestines.

Objective To investigate the expression and clinical significance of intergrin linked kinase (ILK) in pancreatic carcinoma.Methods ILK protein was detected by immunohistochemistry and Western blotting in 60 cases of pancreatic carcinoma and 32 cases of normal pancreatic tissue,and the relationship with the clinicopathological characteristics were analyzed.Results Immunohistochemistry showed ILK was expressed in cytoplasm and membrane of pancreatic carcinoma cells and the positive rate was 65% (39/60),which was significantly higher than 18.75% (6/32) of normal pancreatic tissue(P ＜0.05 ).Western blotting showed the expression of ILK in pancreatic carcinoma tissue was 303933±195116,which was significantly higher than 144613±30074 of normal pancreatic tissue(P＜0.05 ).In pancreatic carcinoma,the expression of ILK was correlated with clinical stage and lymph metastasis(P＜0.05 ),but not correlated with tumor cell differentiation(P＞0.05 ).Conclusions ILK protein was highly expressed in pancreatic carcinoma tissue and it was correlated with the degree of malignancy.

Objective To investigate the role of serum specific IgE(sIgE)antibodies,autoantibody and contact allergens in the pathogenesis of urticaria and the possible correlation among them.Methods The serum sIgE antibodies,autoantibody and contact allergens were tested by IVT kit,autologous serum test-ing and patch testing in145cases of urticaria.Results72.4%(84/116)of patients were positive with spe-cific IgE antibodies,among them13.8%(16/116)were strongly positive.37.7%(20/53)patients were pos-itive with autologous serum testing.In twenty-six patients the above two antibodies were tested.Specific IgE antibodies were found to be strongly positive in4(33.3%)of12patients with negative autologous testing,while in none of14patients with positive autologous testing.Patch testing was performed in21patients with chronic urticaria,90.5%(19/21)patients were positive to15of20allergens tested.Conclusion The pathogenesis of urticaria is complicated,which may include allergy to food,aeroallergens,contact allergens,as well as autoimmunity.Different allergic reactions may be present in the same individual.Contact allergens may be the factor responsible for contact urticaria.

Objective To produce human papillomavirus type 6(HPV-6)virus-like particles with Escherichia coli expression system and study its immunogenicity.Methods HPV-6 L1 gene was inserted into pmkaryotic expression vector pTO-T7 and then expressed in Escherichia coli ER2566.The HPV-6 L1 protein was purified by ammonium sulfate precipitation,ion-exchange chromatography,and hydrophobic interaction chromatography.Then the purified HPV-6 L1 self-assembled into virus-like particle after removing 1,4dithiothreitol(DTr).The morphology of the virus-like particles was investigated with dynamic light scatter and transmission electron microscopy,and the immunogenicity was determined with in vitro pseudownons neutralization as8ay.Results HPV-6 L1 was expressed in soluble form in Escherichia coli.Following the removal of DTT,purified HPV-6 L1 protein could assemble into virus-like particles as 25 am in the radius.And the animal immunization test showed HPV-6 virus-like particles can elite hish titer neutralizing antibodies.Conclusion The bacterially expressed HPV-6 L1 VLP is highly immunogenieity and easy to produce.And it can be good candidate of HPV-6 vaccine.

Objective To investigate the interference of thyroglobulin antibodies ( TgAb ) on the measurement of thyroglobulin ( Tg) by 2 chemiluminescence immunoassays ( CLIAs) .Methods Data of 199 315 individuals with determined TgAb and Tg , including physical checkup subjects , differentiated thyroid carcinoma ( DTC) patients and patients with other diseases , were retrospectively collected in Peking Union Medical College Hospital from November 2012 to April 2015.The correlation between serum Tg level and serum TgAb concentration was analyzed and the positive rate of TgAb in physical checkup subjects was calculated.Furthermore, 290 serum samples with different TgAb concentration were applied in the recovery test by adding in confirmed serum Tg .The correlation between the recovery of confirmed serum Tg and TgAb concentrations was evaluated using Pearson correlation analysis .Results The serum Tg was all decreased with the elevated TgAb concentration in each group of subjects .The positive rate of TgAb was 10.84%(8 416/77 634) in physical checkup subjects .It was higher in females than in males and was increased with age.Recovery test showed that the average recoveries of confirmed serum Tg in TgAb-negative serum were 107.28%(86.30%-117.60%) and 107.94% (85.60%-124.10%) respectively in Roche and Beckman systems.But in TgAb-positive serum samples , the average recoveries in Roche and Beckman systems were 88.59% (35.85% -141.53%) and 95.77% (36.48% -131.78%) respectively, and 12.63%(24/190) and 13.68%(26/190) samples displayed a recovery less than 80%.The recovery rate of confirmed serum Tg showed a significantly negative correlation with elevated TgAb concentration , with r=-0.239 (P=0.001) in Roche and r=-0.251 (P<0.001) in Beckman.Conclusions TgAb-positive serum, especially with high concentration of TgAb , significantly interfered the measurement of Tg .Thus, serum TgAb should be determined together with serum Tg to explore whether there was an interference .To avoid misdiagnosis and inappropriate therapy , clinician should be informed once serum TgAb displayed positive.

BACKGROUND: Accurate serum total thyroxine (TT4) measurement is important for thyroid disorder diagnosis and management. We compared the performance of six automated immunoassays with that of isotope-diluted liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) as the reference method. We also evaluated the correlation of thyroid stimulating hormone (TSH) with TT4 measured by ID-LC-MS/MS and immunoassays. METHODS: Serum was collected from 156 patients between October 2015 and January 2016. TT4 was measured by immunoassays from Abbott (Architect), Siemens (ADVIA Centaur XP), Roche (E601), Beckman-Coulter (Dxi800), Autobio (Autolumo A2000), and Mindray (CL-1000i), and by ID-LC-MS/MS. Results were analyzed using Passing-Bablok regression and Bland-Altman plots. Minimum requirements based on biological variation were as follows: a mean bias of ≤4.5% and total imprecision (CV) of ≤3.7%. RESULTS: All immunoassays showed a correlation >0.945 with ID-LC-MS/MS; however, the slope of the Passing-Bablok regression line varied from 0.886 (Mindray) to 1.23 (Siemens) and the intercept from −12.8 (Siemens) to 4.61 (Mindray). Only Autobio, Beckman-Coulter, and Roche included the value of one in the 95% confidence interval for slope. The mean bias ranged from −10.8% (Abbott) to 9.0% (Siemens), with the lowest value noted for Roche (3.5%) and the highest for Abbott (−10.8%). Only Abbott and Roche showed within-run and total CV ≤3.7%. CONCLUSIONS: Though all immunoassays correlated strongly with ID-LC-MS/MS, most did not meet the minimum clinical requirement. Laboratories and immunoassay manufacturers must be aware of these limitations.
Bias (Epidemiology) ; Diagnosis ; Humans ; Immunoassay ; Mass Spectrometry ; Methods ; Thyroid Gland ; Thyrotropin ; Thyroxine

Objective To investigate the changes of prevalence of hyperuricemia ( HUA) and its correlations with blood glucose and lipid in healthy adults receiving physical examination at Peking Union Medical College Hospital (PUMCH) from 2012 to 2017. Meth-ods An observational approach was adopted for the data analysis.The test results of uric acid (UA),fasting blood glucose (FBG),to-tal cholesterol (TC),triacylglycerol (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C), creatinine (Cr) and Urea of 399 089 cases (206 881 males and 192 208 females) at PUMCH from January 2012 to December 2017 were collected and statistically analyzed.Results The total prevalence of HUA was 17.4% in which the prevalence of males was signif-icantly higher than that of females (25.6% vs 8.5%,χ2＝20 234.850,P＜0.01).During the years of 2012 to 2017,the prevalence of HUA was 26.5%,24.7%,28.6%,23.9%,24.8% and 24.5% in males,and 13.8%,6.3%,7.9%,6.1%,6.2% and 6.8% in females for each year respectively.The prevalence of HUA in males aged 18 to 64 years old was significantly higher than that in the age-matched fe-males (all P＜0.05).However, the prevalence of HUA in males aged≥65 years old was similar to the age-matched females.There was no statistically significant difference of HUA prevalence between males and females aged ≥65 in 2013,2015,2016 and 2017 ( χ2＝1.792,0.017,1.440 and 0.205 respectively;all P＞0.05).The percentages of hyperlipidemia in both males and females of HUA group were higher than those of non-HUA group respectively (all P＜0.01).The percentage of hyperglycemia in males of non-HUA group was higher than that of HUA group,but the percentage of hyperglycemia in females of non-HUA group was lower than that of HUA group ( all P＜0.01).High levels of TC,TG and FBG were risk factors of HUA with increased OR values in increased concentrations of TC,TG and FBG,respectively.Conclusion During the recent 6 years, in healthy adults receiving physiced examination at PVMCH, the preva-lence of male HUA diagnosed was at overall high level,but the prevalence of female HUA was in decreasing and relatively stable trend. Hyperlipidemia and hyperglycemia should be the risk factors of HUA.

Objective To investigate the changes and related factors of maternal thyroid autoantibodies during early pregnancy. Methods Urinary iodine concentration( UIC) , serum thyroid stimulating hormone( TSH) , free thyroxine ( FT4 ) , thyroid-peroxidase antibody ( TPOAb ) , thyroglobulin antibody ( TgAb ) concentrations were determined in 7 190 women during early pregnancy in an iodine-sufficient region of China. Results The prevalence of TPOAb positivity and TgAb positivity were 8. 7% and 12. 0% respectively. The prevalence of overt hypothyroidism and subclinical hypothyroidism increased significantly in group of thyroid antibody positivity. The prevalence of TPOAb positivity and TgAb positivity presented a U-shaped curve, ranging from mild iodine deficiency to iodine excess, especially increased significantly in the group with UIC<100 μg/L. Conclusion Prevalence of thyroid antibodies positivity became higher during early pregnancy. The positive thyroid autoantibodies during pregnancy were significantly associated with maternal hypothyroidism. Both iodine excess and iodine deficiency are risk factors of positive thyroid antibodies.