The Value of the Measurement of Urinary Hyaluronic Acid Levels for the Diagnosis of Bladder Cancer

Objective To investigate the value of the measurement of urinary hyaluronic acid (HA) levels for the diagnosis of bladder cancer and the possibility of replacing ELISA-like assay with radioimmunoassay to detect the levels of urinary HA. Methods Using the ELISA-like assay and radioimmunoassay at the same time to measure the HA levels in the urine specimens from 49 bladder cancer patients, 12 benign bladder tumor patients, 30 other genitourinary disease patients and 20 normal controls. Results There is not much difference between the consequences of the urinary HA levels whether we used the ELISA-like assay or radioimmunoassay to detect every specimen (P>0.05). When we used the results with radioimmunoassay for analysis, we found the levels of urinary HA of bladder cancer patients were 2–4 times than those of the benign bladder tumor patients, other genitourinary disease patients or normal individuals (P<0.01); With 137.5 ngHA/mg protein (113.6±23.9 ng/mg) as a minimum cutoff limit, this assay had a good sensitivity (91.8%) and specificity (91.9%) for the diagnosis of bladder cancer. Its difference in sensitivity meant a lot when compared with urine cytology (48.9%,P<0.01). Conclusion The urinary HA assay is a simple, convenient, noninvasive credible and cheap method with satisfactory sensitivity and specificity for the diagnosis of bladder carcinoma; radioimmunoassay is also a good means to measure the urinary HA levels. Key words Bladder carcinoma - Hyaluronic acid - Urine

梅毒螺旋体明胶凝集试验定量检测在神经梅毒诊疗中的临床意义研究

目的探究梅毒螺旋体明胶凝集试验定量检测(TPPA)在神经梅毒诊疗中的临床意义。方法回顾性分析我院收治的神经梅毒患者160例,分别给予酶免免疫吸附法检测(ELISA)、TPPA,探讨TPPA在神经梅毒诊疗中的应用价值。结果TPPA检验在灵敏度、准确度及特异度(97.42%、96.88%、80.00%)上与ELISA检验相比(64.29%、65.63%、67.11%)显著较高(P<0.05)。结论TPPA检验的敏感度、特异度、准确的较高,适用于梅毒的临床诊断,值得应用与推广。

Observation of Insulin Exocytosis by a Pancreatic β Cell Line with Total Internal Reflection Fluorescence Microscopy

INSULIN secretion was traditionally measured with biochemical and immunological methods such as enzyme linked immunosorbant assay and radioimmunoassay. However, these methods can only tell the amount of insulin secreted; they give no information about the secretion process or mechanism of exocytosis. In recent years, an imaging technique known as total internal reflection fluorescence （TIRF） microscopy has been employed to study insulin secretion.

Incidence of toxoplasmosis in patients with cirrhosis

AIM:It is known that toxoplasmosis rarely leads to various liver pathologies,most common of which is granulomatose hepatitis in patients having normal immune systems.Patients who have cirrhosis of the liver are subject to a variety of cellular as well as humoral immunity disorders.Therefore,it may be considered that toxoplasmosis can cause more frequent and more severe diseases in patients with cirrhosis and is capable of changing the course of the disease.The aim of this study was to investigate the frequency of toxoplasmosis in patients with cirrhosis. METHODS:Serum samples were taken from 108 patients with cirrhosis under observation in the Hepatology Polyclinic of the Gastroenterology Clinic,and a control group made up of 50 healthy blood donors.IFAT and ELISA methods were used to investigate the IgG and IgM antibodies,which had developed from these sera. RESULTS:Toxoplasma IgG and IgN antibody positivity was found in 74 (68.5%) of the 108 cirrhotic patients and 24 (48%) of the 50 people in the control group.The difference between them was significant (P<0.05). CONCLUSION:In conclusion,it was found that the toxopiasma sero-prevalence in the cirrhotic patients in this study was higher.Cirrhotic patients are likely to form a toxoplasma risk group.More detailed studies are needed on this subject.

Inhibition of high-mobility group box 1 expression by siRNA in rat hepatic stellate cells

AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.

False Human Immunodeficiency Virus Test Results Associated with Rheumatoid Factors in Rheumatoid Arthritis

Objective To investigate if immunological factors associated with rheumatoid arthritis(RA) affect the result of human immunodeficiency virus(HIV) screening by electrochemiluminescence immunoassay(ECLIA) and enzyme-linked immunosorbent assay(ELISA). Methods 100 RA cases were enrolled from January 2012 to February 2013 into this study. HIV screening was conducted with ECLIA detecting both HIV-1 p24 antigen, HIV-1 and HIV-2 antibodies, with ELISA and colloidal gold method detecting HIV-1 and HIV-2 antibodies. The samples producing positive results were submitted to the Center for Disease Control for confirmation using Western blotting method. The antibody titers of rheumatoid factors(RF) including RF-IgG, RF-IgM, RF-IgA, and CCP-IgG were analyzed by ELISA. Results The HIV positive-rate determined by ECLIA was significantly higher than that by ELISA and colloidal gold method(P<0.01). The false-positive rate of HIV screening was associated with antibody titers of RF-IgG, RF-IgM, RF-IgA, and CCP-IgG in RA(P<0.01). Conclusion Immunological factors, including RF and anti-CCP antibody, may influence the screening of HIV by ECLIA, producing false-positive result.

Interleukin-2 and Interleukin-6 in Recurrent Genital Herpes Patients

Objective:To study the cellular immunity status of patientswith recurrent genital herpes. Methods: Serum levels of interlukin-2 and its solublereceptor and interlukin-6 were measured by ELISA in 34patients with recurrent genital herpes. Results: Serum levels of IL-2 and IL-6 were significantlydecreased in patients compared to healthy controls (P<0.01),and the level of sIL-2R was significantly increased in patientswith recurrent genital herpes (P<0.01). There were nosignificant differences in all variables amongst patientsregarding relapse stage and remission stage (P>0.05). Conclusion: There was a cellular immune deficiency inpatients with recurrent genital herpes.

High throughput monoclonal antibody generation by immunizing multiple antigens

Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.