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Objective: Survivin is one of the apoptosis inhibitor proteins and is rarely expressed in adult normal tissues. However, survivin expression has been detected in various tumors. In this study, we evaluated the usefulness of urinary survivin/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) ratio as a marker for bladder tumor., Patients and Methods: Urine samples were obtained from 72 patients with bladder tumor, 36 with urinary tract inflammation as controls. Survivin and GAPDH mRNA expression was measured by quantitative real-time PCR assay in urine cells. The GAPDH housekeeping gene was used for normalization of survivin expression. We also analyzed survivin protein levels using urine samples and recombinant protein by western blotting., Results: High expression of survivin was confirmed on the protein level using urine samples of bladder tumor by western blotting. Survivin/GAPDH mRNA ratios of bladder tumor quantified by real-time PCR was significantly higher than those of controls (p=0.001). In pathological stage of bladder tumor, survivin/ GAPDH mRNA ratio of pTis was significantly high compared with pTa and pT1 (p < 0.001, p=0.001, respectively). Grade3 tumors expressed high level of survivin/GAPDH mRNA ratio compared with Grade 1 and Grade 2 tumors (p=0.03). The sensitivity, the specificity and AUC(area under the curve) of survivin/ GAPDH mRNA ratio was 83.3%, 86.1% and 0.898, respectively., Conclusion: Measuring survivin/GAPDH mRNA ratio in urine is non-invasive and high sensitive examination. Therefore, survivin/GAPDH mRNA ratio is useful marker for the detection of bladder tumor, especially to detect carcinoma in situ.

Although liver transplantation (LT) is frequently associated with thrombocytopenia, thrombopoiesis following LT remains to be evaluated. We analyzed platelet counts (PLT), immature Platelet Fraction (IPF), and thrombopoietin (TPO) in 8 patients with LT. PLT and IPF were measured using Sysmex XE-2100. TPO was measured with Human TPO ELISA Kit. In 7 of 8 patients with LT, IPF increased prior to the elevation of platelet counts. In 5 of 7 patients with increased IPF, TPO levels increased simultaneously with IPF, suggesting that IPF may reflect the production of platelets in patients with LT. IPF and TPO might be useful to monitor the platelet production in patients following LT.

The diagnosis of disseminate intravascular coagulation (DIC) was first based on the detection of microthrombi, but it currently involves analysis of hemostatic abnormalities by various laboratory tests and further studies are needed to simplify diagnosis of DIC. There are three diagnostic criteria for DIC developed by the International Society of Thrombosis Haemostasis (ISTH), the Japanese Association for Acute Medicine (JAAM) and the Japanese Ministry Health and Welfare. These diagnostic criteria consist of the scoring system by the same global coagulation tests. From ROC analysis, the ability for diagnosis of DIC is similar among three diagnostic criteria. ISTH diagnostic criteria has high specificity and low sensitivity, but JAAM criteria has low specificity and high sensitivity. Several molecular hemostatic markers such as thrombin antithrombin complex, D-dimer, soluble fibrine (SF) and plasmin-plasmin inhibitor complex have to be adopted for further improvement of diagnostic criteria for DIC. Finally, prospective study for new diagnostic criteria for DIC should be carried out in infectious diseases by molecular hemostatic markers.

Although opportunistic infection including fungal infection is often associated with living donor liver transplantation followed by immunosuppressive therapy, antifungal agents are empirically given to most patients without a definitive diagnosis of fungal infection. Indeed, there is no diagnostic test available, that shows both high sensitivity and specificity for fungal infection. In this study, we developed a polymerase chain reaction(PCR)-based system for rapid diagnosis of fungal infection. This system consisted of two PCR steps: the first PCR for most species of fungi and 4 kinds of nested PCR for Aspergillus or Penicillium (ASP/PEN), Candida glabrata (C. glab), other Candida species including Candida albicans, Candida parapsilosis, Candida tropicalis, and Candida guiliermondii (CAN), and a broad spectrum of fungi (Broad). The newly developed PCR-based system was applied to 28 recipients of with living donor liver transplantation to determine its clinical usefulness in early and differential diagnosis of fungal infection. A total of 514 blood samples from 28 patients ware analyzed. Nested PCR assays were positive in 118 samples from 19 patients: 4 patients (30 samples) with ASP/PEN, 5 patients (29 samples) with C. glab, 12 patients (61 samples) with CAN. All of these samples were positive with nested PCR for Broad as well. Even in samples that were negative on blood culture or ELISA for Aspergillus antigen, nested PCR assays were able to amplify the fungal DNA. Furthermore, all samples positive for Aspergillus antigen tested positive on nested PCR. In conclusion, the PCR-based system we developed was thought to be useful for early diagnosis of fungal infection.

Fibrin/fibrinogen degradation products(FDP) have been measured using serum samples which were specially prepared for the FDP test because of the usage of anti-human fibrinogen antibody for the assay. Since diagnostic criteria for DIC were established by the study group on thrombosis and hemostasis which is supported by the Japanese Ministry of Health and Welfare(JMHW), serum FDP assay have been used as standard methods to diagnose DIC in Japan. Recently, a reagent using an anti-human FDP monoclonal antibody was developed and this has enabled the use of plasma samples for FDP measurement. The comparability, especially of the DIC score, of a new assay, Latex test BL-2 P-FDP, using plasma samples with a conventional assay for serum was investigated. Two sets of DIC scores based on data from the two tests were compared and the correlation was high with 97.5% of the patients being diagnosed with the same DIC status. In four disease groups--DIC, thrombosis, leukemia and solid cancer--high comparability between the two tests was also shown and no significant difference was observed in the correlation coefficient and the slope coefficient between serum and plasma samples. To conclude, it is suggested that "Latex test BL-2 P-FDP" is applicable to the diagnostic criteria for DIC from JMHW without any difficulty.

The differences between reagents of prothrombin time (PT), fibrinogen and fibrin and fibrinogen degradation products(FDP) were examined in patients with disseminated intravascular coagulation (DIC) and without DIC. The sensitivity of the PT ratio for DIC is lowered by the PT reagent with a high international sensitivity index, and the difference between PT reagents was marked. The sensitivity of PT-international normalized ratio (INR) for DIC was higher than that of the PT ratio and the difference between reagents in PT-INR was low. Though the difference between reagents for fibrinogen is slight, the usefulness in diagnosing DIC is also slight. Though the sensitivity of FDP for DIC was good, the difference between FDP reagents was marked. Therefore, standardization of PT and FDP seems to be necessary. Concordance of overt-DIC diagnostic criteria by the International Society of Thrombosis and Haemostasis (ISTH) and DIC diagnostic criteria of Japanese Ministry of Health and Welfare (JMHW) was about 70%, and overt-DIC diagnostic criteria of ISTH seemed to diagnose the typical type of DIC diagnosed by JMHW criteria. Finally, the diagnostic criteria of non-overt DIC are expected to become increasingly important.