Objective This paper aims at establishing a inductively coupled plasma mass spectrometry ( ICP-MS) method for quantification and evaluation of iodine in human urine and serum in routine clinical laboratory .Methods This study was methodology validation research on iodine evaluation using ICP-MS.Ammonia, isopropanol and ultrapure water were mixed at certain ratio to dilute samples in the ratio of 1:10, and then the diluted samples were analyzed by ICP -MS.Re was used as the internal standard.And linearity, lower limit of detection, recovery, precision, accuracy, carryover and stability was evaluated thoroughly .Results of iodine of pregnant women who required iodine tests were retrospectively analyzed to evaluate the status of iodine .Results The method only needs 30s for analysis of one sample .It was sensitive with a lower limit detection of 0.87μg/L, the correlation coefficient was higher than 0.999 9 in ten measurements.The recovery in both serum and urine was approximately 100% (95.3% -109.9%). Based on the NIST standard reference material 3668 comparison, the bias was less than 4%( -0.9% -3.9%).The inter-coefficient variation (CV) for serum iodine and urine iodine was 1.2%-3.0%, 2. 0%-2.9%, respectively;and total CV for serum iodine and urine iodine were 3.0%-3.8%, 4.1%-4.9%, respectively.The mean carryover of this method was 0.03% and iodine was stable for at least one month at -20℃ and 4℃.The urine and serum iodine for pregnant women was (154.8 ±89.7) μg/L (mean ±SD),(75.8 ±21.4) μg/L, respectively.The correlation between urine and serum iodine was 0.21. Conclusion Establishe a rapid and simple ICP -MS method for urine and serum iodine measurement with high accurate and precise in routine clinical laboratory .

OBJECTIVETo develop monoclonal antibodies against the catalytic subunit of human telomerase hTERT for its expression detection of human tumors.

METHODSA dominant epitope in hTERT (peptide hTERT(9))was automatically synthesized based on Fmoc method, and was used to immunize BALB/c mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, and the characterization of which were performed by Western blotting and immunohistochemical staining.

RESULTSAntigenic peptide hTERT(9) was synthesized and confirmed by MALDI-TOF-MS and HPLC analysis. Three hybridoma cell lines secreting anti-hTERT(9) antibodies designated as H4, G8 and A11 were established after primary screening and consequent three rounds of limited dilution. Both of H4 and G8 were IgM, while A11 was IgG1 in isotyping. The competitive assay showed that the antibodies were hTERT(9) specific, and the affinity of G8 was stronger than that of H4 and A11 assayed by affinity ranking. However, in Western blotting, both of H4 and G8 stained an about 123 000 protein band with HeLa and 293 cell extracts but not with normal 2BS cells. Besides, positive staining presented in the nucleus of HeLa, while 2BS was non-reactive immunohistochemically. The sections from paraffin-embedded blocks of 127 cases of human cancer, 40 of precancerous and 19 of benign tumors were in situ stained by G8 antibody, the results showed that the human cancer tissues were 80.31% (102/127) positive in specific nuclear reaction, on the contrary, only a minority of precancerous lesions present weak positive (17.5%, 7/40), and negative in benign tumors (0/19).

CONCLUSIONSThe monoclonal antibodies developed against synthetic peptide were hTERT-specific and could recognize both the native and the denatured form. Thus their use in immunoblotting or immunohistochemistry for detecting the telomerase hTERT expression of cancer cell and tissues was promising.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Binding, Competitive ; Blotting, Western ; methods ; Catalytic Domain ; DNA-Binding Proteins ; Female ; HeLa Cells ; Humans ; Immunohistochemistry ; methods ; Mice ; Mice, Inbred BALB C ; Neoplasms ; enzymology ; pathology ; Telomerase ; chemical synthesis ; immunology

Objective To assess the interference by calcium dobesilatein 7 peroxidase-baseduric acid assays and to determine its clinical significance.Methods In the in vitro experiments, uric acid in pooled serum with final concentrations of calcium dobesilate additions (0, 2, 4, 8, 16, 32, and 64μg/ml) were measured by 7 peroxidase-based assays.Percent Bias (%) was calculated relative to the drug-free specimen.In the in vivo experiments, changes in serum uric acid and calcium dobesilate concentrations were observed before and after calcium dobesilate administration ( baseline, 0 h, 1 h, 2 h, 3 h, 4 h, 6 h ) involunteers.The interference in different assays was assessed compared with LC-IDMS/MS method. Calcium dobesilate levels in 40 specimens from those taking calcium dobesilate were measured by HPLC method.Of the 40 specimens, 10 were selected to analyse the levels of uric acid by both peroxidase and UV measurement method to assess the impact in clinical status.Results In the in vitro study, concentrations of uric acid measured by 7 peroxidase-based assays were reduced by -6.3%to -21.2%compared with drug-free serum, when theconcentration of calcium dobesilate was16μg/ml.In the in vivo study, comparedto UA levels at 0 h, the biasesof serum uric acid determined by peroxidase method after calcium dobesilate administration(1 h, 2 h, 3 h, 4 h, 6 h) were of -3.33%, -6.79%, -7.49%, -6.07%, -4.09%, respectively.The observed uric acid concentrations for 8 participants measured by enzymatic assays were inhibited by -3.75% to -6.89% at 0 hour and by -16.9% to-22.22% at 2 hours relative to the concentrations measured by the LC-IDMS/MS method. Conclusions Calcium dobesilate produced a clinically significant negative interference with uric acid in all peroxidase-based uric acid assays,which may result in false evaluation of uric acid level in clinical status.Significant differences in the degree of interference were observed among the assays.

Chimeric antigen receptor T (CAR-T) cell therapy is an emerging immunotherapy that has allowed for major breakthroughs in the treatment of hematological neoplasms. However, little progress has been made in the treatment of solid tumors, primarily due to the difficulty in homing to tumor tissues by CAR-T cells during treatment. The complex tumor microenvironment and the barrier function of tumor tissues prevent CAR-T cells from contacting tumor cells, thereby preventing them from exerting their antitumor ac-tivity. This review article summarizes not only the progress made in the study of homing disorders of CAR-T cells in the treatment of solid tumors but also the current methods to overcome these disorders.

Gastric cancer (GC) is one of the most resistant malignancy to several treatment strategies. In recent years, the morbidity and mortality of GC has stayed high. Surgical resection remains the main treatment option for GC at present. Although chemotherapy, radiotherapy, immunotherapy, and traditional Chinese medicine are used as adjuvant therapies after surgery, the prognosis and five-year survival rate are still low. Currently, there is no effective method for early diagnosis of GC and thus most patients are diagnosed only when the advanced symptoms appear. Exosomes contain and transfer DNA, RNA, proteins, lipids, and other biological macromole-cules. Several studies have found that exosomes are involved in tumor processes including cell proliferation and metastasis. In particu-lar, the abundant biological macromolecules present in the exosomes reflect the progress of tumor development thereby enabling them as non-invasive diagnostic markers. This provides a new idea for diagnosing GC in early stages. In this review, the contribution of exosomes to GC metastasis and the applications of exosomes in early GC diagnosis are briefly summarized.

Objective To validate the performance of six enzymatic glycated albumin reagents and evaluate their clinical application.Methods The performance of six enzymatic glycated albumin reagents(labled as A,B,C,D,E,F) from Beijing Jiuqiang Co, Beijing Lideman Co,Ningbomeikang Co, Beijing Haomai Co, Sichuan Maike Co and Asahi Kasei Co were assessed on Olympus AU5800 automatic biochemistry analyzer.According to the standard of CLSI,the precision,interference and linear correlation of these reagents were assessed.To assess the accuracy of GA% ,we used GA standard material whose value had been assigned using ID-LC/MS method provided by ReCCS.To do the method comparison and determine the consistency of assay, 50 fresh serum samples of T2DM outpatient and 80 fresh serum samples of apparently healthy people in Jan 2016 were tested using six kits.According to the EP28-A3C protocol, the reference range for GA%was validated in 122 apparently healthy individuals undertaking medical examination from January to February 2016 in PUMC.Results The precision,and the ability of anti-interference of the six reagents were good.The accuracy percentage deviation of six reagents was-19.3%-9.2%.The correlation coefficient of domestic reagents A to E and imported reagents F in the determination of GA% was 0.966-0.999, the average absolute bias was 7.0%-10.4%.The coincidence rate of A-E and F in determining abnormal GA% was between 88.5% and 96.9%.The coincidence rate was increased after switching to the reference range for preliminary clinical evaluation.Conclusion Six GA enzymatic kits used in automatic biochemical analyzer have high precision and strong anti-interference ability.Accuracy still needs to be improved.

A 72-year-old female patient was admitted to our hospital due to chronic cough, expectoration and dyspnea for 10 years,aggravated with intermittent fever for 1 month. She was diagnosed as acute exacerbation of chronic bronchitis,chronic pulmonary heart disease,cardiac function class IV,bilateral lower extremity venous thrombosis and thrombocytopenia. The application of antico-agulant drugs and coagulant drugs in the patient needed to be well weighed with the methods of the bleeding score system combined with clinical assessment of actual risk of bleeding. The interactions of drugs harmful to the patient should be considered and the prognosis of the patient should also be evaluated with careful clinical thought to reduce the patient's risk.

A simple and rapid immunochromatographic test strip incorporating a colloidal gold-labeled recombinant Nsp7 antigen probe was successfully developed for the detection of anti-porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in swine. Recombinant Nsp7 protein of PRRSV labeled with colloidal gold was dispensed on a conjugate pad for use as the detector. Staphylococcal protein A and purified porcine anti-Nsp7 antibodies were blotted on a nitrocellulose membrane to form test and control lines, respectively. A comparison of the strip with standard diagnostic tests, enzyme-linked immunosorbent assays and immunoperoxidase monolayer assay, was also performed. The immunochromatographic test strip was shown to be of high specificity and sensitivity. Furthermore, the strip assay is rapid and easy to perform with no requirement for professional-level skills or equipment. It is suggested that the immunochromatographic test strip can be used to quickly and accurately detect PRRSV antibody and to be suitable for diagnostic purposes in the field.
Antibodies* ; Collodion ; Colloids ; Diagnostic Tests, Routine ; Enzyme-Linked Immunosorbent Assay ; Gold Colloid ; Immunochromatography ; Membranes ; Porcine Reproductive and Respiratory Syndrome* ; Porcine respiratory and reproductive syndrome virus* ; Sensitivity and Specificity ; Staphylococcal Protein A ; Swine

Objective To investigate the effect of restrictive intravenous fluid on the complications and quality of life in the advanced hepatocellular carcinoma patients.Methods Clinical data of Three hundred and fifty-seven cases with advanced hepatocellular carcinoma from Mar 2010 to Mar 2016 was retrospectively analyzed.One hundred and sixty-eight cases were recruited in the restrictive intravenous fluid (RIF) group,and One hundred and eighty-nine cases were involved in the control group.The average volume of intravenous fluid of each day,plasma albumin concentration,splanchnocoel hydrops rate,phlebitis,incidence of vomiting,cancer related pain degree,anxiety degree were compared in the two groups.Results The average volume of intravenous fluid of each day in the RIF group [(720.29 ± 106.84) ml] were much lower than that in the control group [(1 820.36±342.12)ml] (P ＜0.05).The plasma albumin concentration in the RIF group [(35.65 ± 2.21)g/L] were higher than that in the control group [(32.25 ±2.32)g/L] (P ＜0.05).The rate of splanchnocoel hydrops,phlebitis,vomiting,bedsores,and hypstatic pneumonia in RIF group were 6.25％,4.69％,8.59％,3.9％,11.72％ and those in the control group were 13.97％,10.92％,17.47％,10.04％,and 24.45％,respectively (P ＜ 0.05).Moreover,the scores of cancer related pain and anxiety were much lower in the RIF group (5.21 ± 1.09,39.12 ± 5.54) than those in the control group(5.68 ± 1.18,41.56 ± 6.78) (P ＜ 0.01).Conclusions Restrictive intravenous fluid therapy can decrease the cancer associated complications and improved the quality of life in the advanced hepatocellular carcinoma patients.

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