Use of complement regulators,CD35, CD46, CD55, and CD59, on leukocytes as markers for diagnosis of viral and bacterial infections

Several complement regulatory proteins exist on self-cells to prevent damage by the serum complement system. In the present study, we aimed to perform quantitative analysis of membrane-bound complement regulators, CR1 (CD35), MCP (CD46), DAF (CD55), and MIRL (CD59), on peripheral blood neutrophils, monocytes, and lymphocytes from healthy controls (n = 36) and febrile patients diagnosed with either bacterial (n = 21) or viral (n = 26) infections. Our results show that: (a) increased CD35 and CD55 levels on neutrophils and monocytes present potent markers of bacterial infection, (b) increased expression of CD46 on monocytes is an indicator of viral infection, and (c) increased CD59 expression on neutrophils and monocytes is a general infection marker. Additionally, CD19-positive B-lymphocytes represent practically the only lymphocyte population capable of expressing CD35. We further developed two novel clinical flow cytometric markers (indices), specifically, clinical mononucleosis (CM)-INDEX (incorporating CD35, CD55, and CD59 expression on lymphocytes) and clinical bacterial infection (CBI)-INDEX (incorporating CD35 and CD55 expression on neutrophils and lymphocytes), for the effective detection of viral mononucleosis and bacterial infection, respectively. In summary, bacterial and viral infections induce different expression patterns of membrane-bound complement regulators in human leukocytes, which may be effectively exploited in clinical differential diagnosis.     

TT病毒核酸的检测

目的初步测定献血员、慢性乙型肝炎、慢性丙型肝炎及慢性非甲～戊型，非庚型肝炎患者中ＴＴ病毒的感染情况，分析不同ＴＴ病毒感染者血清中ＴＴ病毒开放读码框架２区部分基因序列。方法以套式聚合酶链反应测定ＴＴ病毒核酸，取献血员（ＴＸ２）、慢性乙型肝炎（ＴＸ３）、慢性丙型肝炎（ＴＸ４）及慢性非甲～戊型，非庚型肝炎患者（ＴＸ１）各１例ＴＴ病毒ＰＣＲ阳性产物，以双脱氧核苷酸链终止法测定核苷酸序列。结果检测２０例献血员、２９例慢性乙型肝炎、３１例慢性丙型肝炎及３２份慢性非甲～戊型，非庚型肝炎标本，ＴＴ病毒核酸阳性者分别占５．０％（１／２０）、６．９％（２／２９）、２９．０％（９／３１）和３４．４％（１１／３２）。与Ｎ２２克隆相比，ＴＸ１、ＴＸ２、ＴＸ３与ＴＸ４在核苷酸水平的同源性分别为９５．４１％、９８．４７％、９７．４５％和９７．９６％。结论本组献血员、慢性乙型肝炎、慢性丙型肝炎及慢性非甲～戊型，非庚型肝炎患者中均存在ＴＴ病毒感染者。从本组不同ＴＴ病毒感染者血清中分离出４株ＴＴ病毒序列，其开放读码框架２区部分基因序列高度保守     

Stem cell transplantation in poor-risk chronic lymphocytic leukemia: assessment of post-transplant minimal residual disease using four- and six-color flow cytometry and allele-specific RQ-PCR

A total of 178 bone marrow samples were taken for minimal residual disease (MRD) analysis after 34 stem cell transplantations for poor-risk chronic lymphocytic leukemia, and 86 of them were analyzed in parallel by flow cytometry and allele-specific oligonucleotide-PCR (ASO-PCR). ASO primer was successfully designed for all patients whose frozen diagnosis samples were available. Flow cytometry and ASO-PCR were concordant, i.e. both either positive or both negative, in 78% of the analyses. Flow cytometry did not detect MRD in any of the samples that were PCR-negative cases. In contrast, ASO-PCR detected MRD in samples that were negative for MRD by flow cytometry in 22% of the analyses. In one patient, the immunophenotype but not the IgV_H gene sequence had changed during a course of the disease, and MRD could not be followed by flow cytometry. In the remaining cases, the discrepancy was due to a higher sensitivity of ASO-PCR. Autologous stem cell transplantation resulted in clinical complete response in 87% (20/23) of the patients. By flow cytometry, 35% (8/23) of autotransplanted patients became MRD-negative, but only 12.5% (2/16) PCR-negative (sensitivity of ASO-PCR <0.001 and <0.01, respectively). All allotransplanted patients achieved or maintained hematological CR, and five out of nine patients (56%) became PCR-negative (sensitivity of PCR between <0.001 and <0.003), two of them having non-myeloablative conditioning. None of the patients who became PCR-negative after allogeneic transplantation have relapsed.     

Clinical features and predictors of diphtheritic cardiomyopathy in Vietnamese children

BACKGROUND: Despite the availability of antitoxin and antibiotics, the mortality rate for diphtheria remains high, mostly because of cardiac complications. METHODS: During 1 year, 154 Vietnamese children with diphtheria admitted to a referral hospital were studied prospectively with clinical examination, including a simple pseudomembrane score, 12-lead and 24-hour electrocardiography, measurement of serum cardiac enzyme levels, and estimation of troponin T levels. RESULTS: Thirteen children had diphtheritic cardiomyopathy on admission, and 19 developed it subsequently. Twelve children (8%) died. The combination of pseudomembrane score of >2 and bull neck predicted the development of diphtheritic cardiomyopathy, with a positive predictive value of 83% and a negative predictive value of 93%. Administration of 24-hour electrocardiography on admission improved the ability to predict diphtheritic cardiomyopathy by 57%. Fatal outcome was best predicted by the combination of myocarditis on admission and a pseudomembrane score of >2. Of the cardiac enzyme levels measured, an elevated aspartate aminotransferase level was the best predictor. The presence of troponin T identified additional children with subclinical cardiac damage. CONCLUSIONS: The development of diphtheritic cardiomyopathy can be predicted by means of simple measures.     

Immunohistochemical localization of types I and III collagen and fibronectin in the dentine of carious human teeth

Reparative dentine formed as a response to caries was mostly type I collagen similar to that of normal dentine. The predentine related to reparative dentine reacted positively with antisera to types I and III collagen and to fibronectin. Normal odontoblasts and their processes reacted positively both with types I and III collagen antibodies. Fibronectin was related to odontoblasts and their processes pericellularly. Odontoblasts appeared not to lose totally their developmental ability to synthesize type III-like molecules after maturation. Pulp fibroblasts reacted positively both with types I and III antibodies as well as antifibronectin. The cell-free and cell-rich zones revealed a dense layer of fibres reacting positively with type III collagen and fibronectin antibodies. The width of these zones were reduced in relation to reparative dentine.     

A prospective randomized trial of the surgical blood order equation for ordering red cells for total hip arthroplasty patients

BACKGROUND: The majority of crossmatched blood is for surgical patients, and most of it is never transfused. An alternative system for ordering red cell (RBC) units, called the surgical blood order equation (SBOE), which incorporates specific patient variables for surgical patients, has been developed. STUDY DESIGN AND METHODS: A prospective double-blind randomized trial compared the SBOE with the maximal surgical blood order schedule (MSBOS) system for ordering allogeneic RBC units in 60 patients undergoing total hip arthroplasty. Autologous RBCs were available for none of the patients. RESULTS: There were no differences in patient demographic, surgical, or laboratory variables at any time. The median number (range) of allogeneic RBC units ordered was 2 (2-3) for the MSBOS and 0 (0-3) for the SBOE (p<0.0001). The SBOE ordered the correct number of RBC units for 58 percent of patients, while the MSBOS did so for 7 percent (p<0.0001). The SBOE had a lower crossmatch-to-transfusion ratio than the MSBOS (0.83 vs. 4.12). Costs were also lower with the SBOE. CONCLUSION: Incorporation of patient factors in the use of the SBOE system resulted in increased efficiency of blood-ordering practices for total hip arthroplasty.     

Ultrastructural characteristics of cells in human wound collected by Cellstic device

The differentiating human wound cells and intercellular matrix was studied by electron microscopy. The cells were collected with a special wound drain containing a piece of cellulose sponge. Although fibroblastic cells could be detected as early as 20 hr after operation, the main cell population consisted of inflammatory cells. The fibroblastic cells were classified into three types according to the stage of ultrastructural differentiation. Some cells of different types were in close contact by membrane apposition to each other and to the sponge material. However, no specialized junctions were seen. Organized collagen fibers were not seen during the observation period despite the well developed organelles for protein synthesis in the fibroblastic cells. The close contacts of normal cells with the sponge pore walls indicated good compatibility with human tissue and safe use for clinical experiments in routine operations.