A new staging system for multiple myeloma (MM) has utilized serum concentrations of beta 2-microglobulin (S beta2 M) and albumin as important prognostic factors for survival. Since S beta2 M is an indicator of glomerular filtration rate, we compared the prognostic values of S beta2 M and 24-hr urinary creatinine clearance (Ccr) in patients with MM. We retrospectively reviewed the records of 170 MM patients from January 1996 to November 2003 whose 24-hr urinary Ccr was available at the time of diagnosis. We found that pretreatment S beta2 M was inversely related to Ccr (Spearman's correlation coefficient=-0.787). In univariate analysis, the hazard ratio (HR) of death was 1.043 (p<0.001) for S beta2 M and 0.985 (p<0.001) for Ccr. Multivariate analysis showed that S beta2 M (HR 1.030, p=0.010) and Ccr (HR 0.993, p=0.059) were significant prognostic factors in patients' survival. In conclusion, 24-hr urinary Ccr may be utilized for staging of patients with MM.
beta 2-Microglobulin/*blood ; Survival Analysis ; Retrospective Studies ; Prognosis ; Neoplasm Staging/methods ; Multivariate Analysis ; Multiple Myeloma/drug therapy/metabolism/*pathology ; Creatinine/*urine

BACKGROUND: An increased level of beta2-microglobulin (beta2M) is seen in diseases such as lymphoproliferative diseases, renal diseases, solid tumors, liver diseases, certain viral infection, or chronic inflammatory diseases, etc. In this study, we evaluated a quantitative beta2M assay for precision and linearity using an automated turbidimetric immunoassay (TIA) by Hitachi 7600-110 (Hitachi High Technologies Co., Japan). The TIA of beta2M was compared with a chemiluminescent immunoassay (CLIA) by Immulite 2000 (Diagnostic Products Corporation, USA). METHODS: Two TIAs, the Hitachi 7600-110 with Roche reagent (Roche-TIA) and the Hitachi 7600- 110 with HBI reagent (HBI-TIA), were evaluated for within-run precision, within-day precision, betweendays precision, and linearity. With 68 serum samples, two TIAs were compared with Immulite 2000 using DPC reagent (DPC-CLIA). These data were analyzed by Passing-Bablok analysis and Bland- Altman analysis. RESULTS: The coefficients of variation (CVs) of within-run precision, within-day precision, and between- days precision were less than 7% in all groups. The linearity tests of the two TIAs were maintained well (Roche-TIA: R2=0.9952; HBI-TIA: R2=0.9946). The comparison study indicated good correlations (Roche-TIA/DPC-CLIA: r=0.9738, y=0.9625x-0.0375; HBI-TIA/DPC-CLIA: r=0.9725, y= 1.1000x-0.3100). In Bland-Altman analysis, less than 2SD differences were observed between the two groups. CONCLUSIONS: Both Roche-TIA and HBI-TIA showed a good precision and excellent correlations with DPC-CLIA; therefore, TIA could be used in the routine laboratory to determine a quantitative analysis of beta2M.
Aged ; Chemiluminescent Measurements ; Enzyme Multiplied Immunoassay Technique ; Female ; Humans ; Immunoassay/*methods ; Male ; Middle Aged ; Nephelometry and Turbidimetry ; Reagent Kits, Diagnostic ; Regression Analysis ; Sensitivity and Specificity ; beta 2-Microglobulin/*analysis

BACKGROUND/AIMS: Serum ferritin is a marker of acute phase reactions and iron storage. In addition, hematologic malignancies are associated with elevated serum ferritin levels. Other studies have suggested that ferritin is a surrogate for advanced disease and has an impact on relapse, because elevated serum ferritin predicts overall survival (OS) and relapse-free survival following autologous stem cell transplantation for lymphomas. METHODS: We studied 89 consecutive patients with newly diagnosed multiple myeloma to determine the value of serum ferritin in comparison with known prognostic factors. RESULTS: The OS in the elevated serum ferritin group (> or =300 ng/mL) was shorter than that in the normal serum ferritin group (<300 ng/mL, p<0.001) after a median follow-up of 25 months. In univariate analysis, elevated ferritin was correlated with poor survival in the patients (relative risk [RR], 2.588; 95% confidence interval [CI], 1.536 to 4.358; p<0.001). Furthermore, multivariate analysis showed that elevated serum ferritin was an independent predictor of mortality in patients with multiple myeloma (RR, 2.594; 95% CI, 1.403 to 4.797; p=0.002). CONCLUSIONS: The serum ferritin can a prognostic parameter of survival as well as disease activity in patients with multiple myeloma.
Adult ; Aged ; Aged, 80 and over ; C-Reactive Protein/analysis ; Female ; Ferritins/*blood ; Humans ; Male ; Middle Aged ; Multiple Myeloma/*blood ; Prognosis ; Proportional Hazards Models ; beta 2-Microglobulin/blood

OBJECTIVETo use the data of occupational epidemiology to estimate the benchmark dose (BMD) of renal dysfunction induced by lead.

METHODSBlood lead was considered as an exposure biomarker, while urinary total protein (TP), urinary beta(2)-microglobulin (beta(2)-MG) and urinary N-Acetyl-beta-D-glucosaminidase (NAG) were considered as effect biomarkers reflecting the damage of renal function. The dichotomized (binary) data was used as effect endpoints. The BMD and BMD lower limit (BMDL) of blood lead were estimated at the 10% benchmark response using BMDS version 1.3.1.

RESULTSThere was an increased prevalence of hyper-TP-uria, hyper-beta(2)-MG-uria and hyper-NAG-uria with an increasing blood lead concentration. There was obviously dose-response relationship between blood lead and TP, beta(2)-MG and NAG, respectively. The BMD and BMDL of blood lead affecting renal function were estimated to be 323.6 - 754.3 microg/L and 274.2 - 541.5 microg/L. The BMDL of blood lead was ranged from low to high as NAG, TP and beta(2)-MG. The urinary NAG activity might be served as a sensitive biomarker in detecting early renal dysfunction.

CONCLUSIONIt should be feasible to use the BMD approach to set up the reference dose (RfD) and reference concentration (RfC). BMD approach might provide a new and better way for setting up the RfD/RfC.

Acetylglucosaminidase ; urine ; China ; epidemiology ; Clinical Chemistry Tests ; methods ; standards ; Humans ; Lead ; blood ; Lead Poisoning ; blood ; epidemiology ; urine ; Occupational Exposure ; analysis ; Prevalence ; Proteinuria ; urine ; beta 2-Microglobulin ; urine

OBJECTIVEThis review focuses on the current knowledge on the implication and significance of beta 2 microglobulin (&beta;2M), a conservative immune molecule in vertebrate.

DATA SOURCESThe data used in this review were obtained from PubMed up to October 2015. Terms of &beta;2M, immune response, and infection were used in the search.

STUDY SELECTIONSArticles related to &beta;2M were retrieved and reviewed. Articles focusing on the characteristic and function of &beta;2M were selected. The exclusion criteria of articles were that the studies on &beta;2M-related molecules.

RESULTS&beta;2M is critical for the immune surveillance and modulation in vertebrate animals. The dysregulation of &beta;2M is associated with multiple diseases, including endogenous and infectious diseases. &beta;2M could directly participate in the development of cancer cells, and the level of &beta;2M is deemed as a prognostic marker for several malignancies. It also involves in forming major histocompatibility complex (MHC class I or MHC I) or like heterodimers, covering from antigen presentation to immune homeostasis.

CONCLUSIONSBased on the characteristic of &beta;2M, it or its signaling pathway has been targeted as biomedical or therapeutic tools. Moreover, &beta;2M is highly conserved among different species, and overall structures are virtually identical, implying the versatility of &beta;2M on applications.

Antigens, CD1 ; physiology ; Hemochromatosis Protein ; analysis ; Histocompatibility Antigens Class I ; physiology ; Humans ; Receptors, Fc ; physiology ; beta 2-Microglobulin ; blood ; chemistry ; deficiency ; physiology

This study was aimed to construct the soluble HLA-A*0201-PR1 complex for preparation of HLA-A*0201-PR1 tetramer. The recombinant HLA-A*0201-BSP (BirA substrate peptide) fusion protein as heavy chain and beta(2)-microglobulin (beta(2) m) as light chain were expressed highly as insoluble aggregates in Escherichia coli and then purified with gel filtration, and the final purity reached above 90%. The two subunits were refolded to form an HLA-A*0201-peptide complex by dilution method in the presence of an antigenic peptide PR1, a HLA-A2-restricted peptide from proteinase 3 (aa 169 - 177, VLQELNVTV). Refolded HLA-A*0201-PR1 complex was biotinylated using a BirA enzyme and purified by anion exchange chromatography on a Q-Sepharose (fast flow) column. The extent of reconstitution of the HLA-A*0201-PR1 complex was analyzed by HPLC gel filtration. The refolded and biotinylated products were detected by Western blot and ELISA with monoclonal antibody BB7.2 that recognized the natural conformations of HLA-A2 and streptavidin. The results showed that the refolded complex was composed of HLA-A*0201-BSP aggregate, HLA-A*0201-PR1 complex and beta(2) m, and reconstitution yields of 18% with PR1 was obtained. Refolded HLA-A*0201-PR1 complex could be confirmed by practical immunological method and biotinylated efficiently. It is concluded that the refolding and biotinylation of HLA-A*0201-PR1 complex is successfully obtained. This work provides the basis for the preparation of HLA-A*0201-PR1 tetramer.
DNA Primers ; genetics ; Escherichia coli ; genetics ; HLA-A Antigens ; analysis ; biosynthesis ; genetics ; HLA-A2 Antigen ; Humans ; Oligopeptides ; genetics ; metabolism ; Protein Binding ; Protein Folding ; Recombinant Fusion Proteins ; analysis ; biosynthesis ; beta 2-Microglobulin ; biosynthesis ; chemistry ; genetics

BACKGROUNDAcute renal failure (ARF) after liver transplantation is associated with high mortality and morbidity. Early therapeutic or preventive intervention is hampered by the lack of early effective prognostic factors. Recent studies indicated that serum levels of cystatin C and beta2-microglobulin (beta2 MG) as well as urinary beta2 MG and N-acetyl-beta-D-glucosaminidase (NAG) would increase in patients with early and mild renal impairment. In this study, these factors were detected during the different stages in patients who accepted orthotopic liver transplantation (OLT), and their feasibilities to predict early ARF after OLT were also analyzed.

METHODSSixty patients with normal blood urea nitrogen (BUN) and serum creatinine (SCr) who received modified piggyback liver transplantation without veno-venous bypass were prospectively studied. Blood samples were drawn from patients for the determination of serum beta2 MG (n = 60), SCr (n = 60) and serum Cystatin C (n = 39) at following 5 intervals: before operation (T0), 20 minutes before anhepatic phase (T1), 25 minutes in anhepatic (T2), 60 minutes after reperfusion (T3) and at the end of operation (T4). Urinary beta2 MG (n = 60) and NAG (n = 60) were also examined at following 3 intervals: before operation (T0), 60 minutes after reperfusion (T3) and at the end of operation (T4). According to the Rimola A criteria of ARF in 24 hours after operation, all the patients were divided into two groups: ARF group and non-ARF group. The data were statistically analyzed to evaluate the feasibiliy of regarding these factors as prognostic factors for early ARF after liver transplantation in patients with normal SCr and BUN before operation.

RESULTSTen of sixty cases showed ARF (16.7%). The Logistic regression analysis showed that the levels of serum and urinary beta2 MG as well as serum cystatin C before operation were correlated with early ARF after liver transplantation (P < 0.05), while only serum levels of cystatin C and Cr at the end of operation correlated with early ARF (P < 0.05, P < 0.01) after liver transplantation. The serum beta2 MG, Cystatin C, SCr and urinary beta2 MG levels in ARF group were much more higher than that in non-ARF group (P < 0.05, P < 0.01). There were significant differences between the correct and false predictive positive ratios of serum cystatin C, serum and urinary beta2 MG levels before operation (P < 0.05, P < 0.01), while only SCr showed significant difference between these groups at the end of operation (P < 0.01).

CONCLUSIONSThe results revealed that there was potential renal damage among those patients who demonstrated normal SCr and BUN before operation, and that liver transplantation could aggravate this damage and causing ARF. Here we provided the prognostic values of serum Cystatin C, beta2 MG, urinary beta2 MG and NAG in patients with early acute renal failure after liver transplantation.

Acetylglucosaminidase ; urine ; Acute Kidney Injury ; blood ; diagnosis ; urine ; Adult ; Blood Urea Nitrogen ; Cystatin C ; blood ; Female ; Humans ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; blood ; diagnosis ; urine ; Predictive Value of Tests ; Prognosis ; beta 2-Microglobulin ; analysis ; blood ; urine

OBJECTIVETo observe the effect of long-term external use of Goupi Gao on renal function and lead accumulation in rats.

METHODRats were externally administered with Goupi Gao at different doses (7, 3.5 and 1.75 g x kg(-1)) for 90 d. At 45 days and 90 days after administration, the renal indicator, levels of blood urea nitrogen (BU) and creatinine (Cr) in serum, beta2-microglobulin (beta2-MG) and N-acetyl-beta-D-glucosaminidase (NAG) in urine were determined. Lead content in kidneys was detected by atomic absorption spectrometry.

RESULTA 90-day administration with Goupi Gao significantly enhanced the renal indicator, levels of NAG in urine and lead content in renal, when compared with the normal rats. However, the levels of BUN and beta2-MG as well as renal pathology in Goupi Gao treated rats were not obviously changed.

CONCLUSIONConsecutive administration of Goupi Gao for 90 days can increase the renal indicator and levels of NAG in urine, enhance the accumulation of lead in renal, but with no effect on excretory function of kidneys and organic changes.

Acetylglucosaminidase ; urine ; Animals ; Blood Urea Nitrogen ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Female ; Kidney ; drug effects ; metabolism ; pathology ; Lead ; analysis ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry, Atomic ; beta 2-Microglobulin ; urine

OBJECTIVETo define the mechanism of acute hepatitis in non-human primates after liver directed gene therapy.

METHODSDifferences in immune response exhibited by 8 rhesus monkeys receiving adenovirus (Ad) or lipofectamine-mediated gene transfer by various routes, the time course, and the nature of the specific immune responses to both adenoviral vectors and transgene products were studied using HE staining (H&E) and immunohistochemical staining.

RESULTSThe monkeys developed mild to moderate acute hepatitis 1 to 3 weeks after intravenous or intrabiliary injection of first generation replication-defective adenoviruses carrying the Escherichia coli lacZ gene. This was accompanied by adenovirus-mediated T-cell proliferation and neutralizing antibodies to the adenovirus. Increased numbers of CD3(+), CD4(+) and CD8(+) T-lymphocytes were detected in the diseased livers, while B-lymphocytes were absent. Hepatocytes demonstrated increased expression of beta 2-microglobulins (beta 2-MG) and HLA-DR antigens in the plasma membranes. The development of acute hepatitis and the accompanying immune abnormalities were delayed in immunosuppressed monkeys until after the discontinuation of immunosuppressive therapy. The monkeys infused with Ad. CMVluc showed more significant and longer durations of hepatitis than the monkeys infused with adenoviruses carrying the lacZ gene. Lipofectamine-mediated gene transfer was inefficient. There was neither lacZ expression nor significant immune response in the liver of monkeys infused with lipofectamine via the portal vein or the common bile duct.

CONCLUSIONImmune response to the hepatocytes in liver directed gene therapy is MHC class I restricted and T-cell mediated. Both adenoviral vectors and foreign genes are related to the liver damage. Mild to moderate hepatic inflammation seen with the E-1 deleted vector is reversible. Immunosuppression regimens may prolong transgene expression and delay the development of acute adenoviral hepatitis.

Acute Disease ; Adenoviridae ; genetics ; Adenoviridae Infections ; genetics ; Animals ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; DNA, Recombinant ; administration & dosage ; genetics ; Gene Transfer Techniques ; HLA-DR Antigens ; analysis ; Hepatitis, Animal ; genetics ; virology ; Immunohistochemistry ; Liver ; chemistry ; metabolism ; pathology ; Macaca mulatta ; beta 2-Microglobulin ; analysis

Some circulating cancer cells in the blood play a central role in the metastatic process and may have a major influence on patient progress. Their numbers can be very small and techniques for their detection need to be both sensitive and specific. Polymerase chain reaction (PCR) has been successfully used to detect small numbers of tumor cells in cancer. We used a reverse transcriptase-polymerase chain reaction (RT-PCR) to detect circulating breast cancer cells in venous blood samples before operations and assessed cytokeratin-19 (CK-19) and cytokeratin-20 (CK-20) as target mRNA markers in the blood of healthy donors (n=6) and breast cancer patients (n=30) with American Joint Committee on Cancer stages 0 to IIIa. CK-19 mRNA was expressed in all blood samples of healthy donors and patients. But CK-20 was the only mRNA marker not detected in the blood from healthy donors. Seven of 30 (23%) venous blood isolates of breast cancer patients yielded a CK-20 mRNA with positive results. There was no correlating CK-20 mRNA expression with stage and axillary lymph node status. In conclusion, CK-19 showed no diagnostic value as a mRNA marker in the detection of circulating cancer cells by RT-PCR assay because this was expressed in the blood of healthy donors. CK-20 mRNA was an useful marker to detect circulating cancer cells in breast cancers.
Breast Neoplasms/pathology* ; Breast Neoplasms/genetics* ; DNA Primers ; Female ; Gene Expression Regulation, Neoplastic ; Genetic Markers ; Human ; Intermediate Filament Proteins/genetics ; Keratin/genetics ; Neoplasm Circulating Cells* ; RNA, Messenger/analysis ; RNA, Neoplasm/analysis* ; Reverse Transcriptase Polymerase Chain Reaction/methods* ; Sensitivity and Specificity ; beta 2-Microglobulin/genetics