The Podocyte Count Detected by an Improved Immunocytochemical Method has Higher Diagnostic Efficiency than Enzyme-Linked Immunosorbent Assay and Serum Cystatin C to Evaluate the Early Stage Damage of Glomerular.

BACKGROUND: In renal diseases, earlier injury to glomerular may lead to the abscission of podocyte. The number of podocyte in urine may reflect the severity of glomerular damage. Podocalyxin (PCX) was considered as a podocyte marker. Many methods were used to detect podocyte. Applications of these methods were limited by tricky, expensive, and low accuracy. Here, we improved an immunocytochemical method to count the number of podocyte in urine. METHODS: In this study, we counted the numbers of podocyte in urine by our improved method and detected the PCX levels in urine by enzyme-linked immunosorbent assay (ELISA) in glomerulopathy patients and healthy controls. The serum levels of cystatin C (CysC), blood urea nitrogen (BUN), creatinine (CR), uric acid (URIC), and ß2-Microglobulin (ß2-MG) in all subjects were detected. Correlation analysis and diagnostic efficiencies comparisons among immuocytochemical method, ELISA, and the biochemistry index were also performed. RESULTS: The podocyte counts in patients were significantly higher than controls. Podocytes counts positively correlated with PCX concentrations and the serum CysC. The podocyte count had higher diagnostic efficiency than PCX concentrations detected by ELISA and serum CysC. CONCLUSIONS: The podocyte count detected by our improved method had higher diagnostic efficiency than ELISA and serum CysC.

The Usage of Separating Gel Vacuum Tube with Different IgG Concentration.

BACKGROUND: Observing serum IgG concentration on the distribution of serum, blood cells, and separation gel after centrifugation in different separation gel vacuum tubes. METHODS: 3 mL venous blood was collected in each of two separation gel procoagulant vacuum tubes: BD Vacutainer SST II(3.5ml, 75×13 mm) and BD Vacutainer SST(5ml, 100×13 mm). After complete solidifaction, both tubes were centrifuged at 2000g for 10 minutes. The distribution of serum, blood cells, and separation gel in the vacuum tube was observed. The immunoglobulin concentration was detected using the special protein analyzer Siemens BNII. RESULTS: 1. In the group of BD Vacutainer SST II where the IgG concentration exceeded 50g/L but less than 122g/L: The serum was located below the separation gel and was distributed in three layers: separation gel, serum, and blood cells. 2. In the group of BD Vacutainer SST where the IgG concentration exceeded 50g/L but less than 122g/L: The serum was located above the separating gel, and was distributed in three layers: serum, separation gel, and blood cells. 3. Increases in IgA and IgM serum concentration did not cause the separation gel inversion. CONCLUSIONS: Increases in serum IgG were positively correlated with the concentration of total protein. The rising of serum IgG caused the floating of separation gel after centrifugation. The BD Vacutainer SST was more suitable for clinical blood sample collection.

Interleukin-18 synergism with interleukin-2 in cytotoxicity and NKG2D expression of human natural killer cells.

Natural killer (NK) cells play an important role in anti-tumor immunity. Interleukin (IL)-18 is an immunoregulatory cytokine that induces potent NK cell-dependent anti-tumor responses when administrated with other cytokines. In this study, we explored the effects of combining IL-18 and IL-2 on NK cytotoxicity as well as expression levels of the NK cell receptor NKG2D in vitro. Freshly isolated PBMCs were incubated for 48 h with IL-18 and IL-2, then CD107a expression on CD3-CD56+ NK cells was determined by three-colour flow cytometry to evaluate the cytotoxicity of NK cells against human erythroleukemia K562 cells and human colon carcinoma HT29 cells. Flow cytometric analysis was also employed to determine NKG2D expression on NK cells. The combined use of IL-18 and IL-2 significantly increased CD107a expression on NK cells compared with using IL-18 or IL-2 alone, suggesting that the combination of these two cytokines exerted synergistic enhancement of NK cytotoxicity. IL-18 also enhanced NKG2D expression on NK cells when administered with IL-2. In addition, blockade of NKG2D signaling with NKG2D-blocking antibody attenuated the up-regulatory effect of combining IL-18 and IL-2 on NK cytolysis. Our data revealed that IL-18 synergized with IL-2 to dramatically enhance the cytolytic activity of human NK cells in a NKG2D-dependent manner. The results appear encouraging for the use of combined IL-18 and IL-2 in tumor immunotherapy.

Association of estrogen receptor alpha gene polymorphisms with cytokine genes expression in systemic lupus erythematosus.

AIM: To analyze the association of estrogen receptor alpha (OR alpha) gene polymorphisms with cytokine genes expression in patients with systemic lupus erythematosus (SLE) and controls. METHODS: Genomic DNA was extracted and polymorphisms of XbaI, Ukrainian (XX, Xx, or xx genotype) and PvuII (PP, Pp, or pp) in intron 1 of OR alpha gene were detected by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) method. The messenger RNA (mRNA) levels of interleukin (IL)-10, IL-4, interferon (IFN)-gamma, and IL-2 were assessed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: In patients with SLE with PpXx genotype, IL-10 and IL-4 mRNA expression was higher (P < 0.001 and P = 0.013, respectively), while in patients with SLE with Ppxx genotype IFN-gamma and IL-2 mRNA expression was lower than in controls (P < 0.001). There was no significant difference in mRNA expression of 4 cytokines among controls with various genotypes. CONCLUSION: OR alpha gene polymorphism may be associated with the expression of IL-10, IL-4, IL-2, and IFN-gamma in patients with SLE.