OBJECTIVETo search a method for showing dust granules in phagocytes.

METHODSThe autopsy samples of 10 cases who died of non-pulmonary diseases(group A), 6 cases with silicosis(group B) and 5 cases of dead fetus(group C) were collected. The samples of lung, spleen and lymphnodes were stained by methods of (1)HE staining; (2)Warthin-Starry staining(W-S staining) and modified W-S staining; (3) CD68 (as a first antibody) immunohistochemical staining. The ultrastructure and chemical component of the dust granules in the pulmonary phagocytes(dust cells) of group A(3 cases) and group B(3 cases) were observed and analyzed by transmission electron microscope(TEM) and analytic electron microscope(AEM).

RESULTSIn group A and B smaller or minute dust granules which could not be shown by HE staining method were clearly shown by W-S staining and modified W-S staining methods. There were no positive granules found in group C. The CD68 marker of the dust cells was positive. The size, shape, density and chemical elements of the dust granules of group A were different from those of group B under TEM and AEM, excluding bacteria and other intracellular contents.

CONCLUSIONThe dust granules in the phagocytes could be shown by W-S staining method, its staining effect for minute dust granules is superior to general HE staining. However, the determination of chemical elements of dusts must depend on AEM.

Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Dust ; Humans ; Microscopy, Electron ; Phagocytes ; ultrastructure ; Silicosis ; pathology ; Staining and Labeling ; methods

OBJECTIVETo explore the clinicopathologic features, immunophenotype, differential diagnosis and gene mutation status of the Erdheim-Chester disease (ECD).

METHODSClinical and pathologic findings of 3 ECD cases were examined by gross, microscopic, immunohistochemical methods and BRAF V600E mutation. Related literatures were reviewed.

RESULTSTwo male patients and one female patient presented clinically with multiple skin nodules, bone pain and bony lesions by imaging study. Microscopically, the lesions were composed of spindle-shaped fibroblasts, foamy histiocytes and scattered Touton-type giant cells embedded in reactive fibrous tissue. Lymphocytes, plasma cells, and multinucleated giant cells were also found. Immunohistochemically, all histiocytes were positive for CD68, none of which expressed CD1a, although 2 cases focally expressed weak S-100 stain. In 2 cases,BRAF V600E mutation was detected.

CONCLUSIONSECD is a rare disease of xanthogranulomatous histiocytosis.Its diagnosis relies on pathological and immunohistochemical findings, but correlation with clinical information, especially radiographic findings should be performed.No effective treatment of the disease is currently available.

Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Diagnosis, Differential ; Erdheim-Chester Disease ; genetics ; immunology ; pathology ; Female ; Humans ; Male ; Mutation ; S100 Proteins ; analysis ; Treatment Outcome

To characterize cellular responses during hepatic regeneration, we examined 13 explant livers and 5 liver allografts by immunohistochemistry for cytokeratin 7, HepPar1, CD68, alpha-smooth muscle actin (alpha-SMA) and proliferating cell nuclear antigen as well as reticulin and Masson-trichrome staining. Within a week after liver damage, elongated CD68-positive cells were detected along the border of necrotic area. The number of alpha-SMA-positive cells was slightly increased along the sinusoids. Ductular proliferation or fibrosis was negligible. After one or two weeks, the size and number of CD68-positive cells were markedly increased. alpha-SMA-positive cells increased in number within lobules and portal tracts. Ductular proliferation occurred predominantly at the limiting plate or along the border of necrotic areas. After one month, necrotic parenchyma was replaced by many ductules, CD68-positive cells, alpha-SMA-positive cells. Nodules of regenerating hepatocytes and irregular fibrosis were diffusely present. Other nonparenchymal cells were not significantly changed. These observations indicate that chronological interaction between nonparenchymal and parenchymal cells occur during the course of human hepatic regeneration and suggest extensive porto-periportal fibrosis more than a few months after the onset of fulminant hepatitis is a major indicator of chronic functional impairment necessitating liver transplantation.
Actins/analysis ; Antigens, CD/analysis ; Antigens, Differentiation, Myelomonocytic/analysis ; Human ; Immunohistochemistry ; Keratin/analysis ; Liver/*cytology ; *Liver Regeneration ; Proliferating Cell Nuclear Antigen/analysis

OBJECTIVETo study the clinical significance of CD163 in the diagnosis and the evaluation of severity and prognosis of childhood hemophagocytic lymphohistiocytosis (HLH).

METHODSNinety-four children were classified into Epstein-Barr virus (EBV)-positive (n=55) and EBV-negative groups (n=39; control group). The EBV-positive group was subgrouped into infectious mononucleosis (IM; n=47) and HLH (n=8). Serum levels of soluble CD163 were measured using ELISA. Expression of CD163 on mononuclear cells was detected by flow cytometry.

RESULTSThe serum levels of soluble CD163 were>10 000 ng/mL in all eight HLH patients (>30 000 ng/mL in 3 cases). The mean serum levels of soluble CD163 in the HLH group were significantly higher than in the control and IM groups (P<0.05). The serum levels of soluble CD163 in EBV-positive children were positively correlated with EBV-DNA copies and serum levels of ferritin and LDH, but were negatively correlated with white blood cell count, neutrophil count, hemoglobin and platelet count. The follow-up after treatment for three HLH patients showed that serum levels of soluble CD163 were significantly reduced, but the soluble CD163 levels rebounded in one patient who was complicated by fungal pneumonia infection.

CONCLUSIONSThe levels of serum soluble CD163 may be related to the severity in children with HLH. The EBV-positive children with soluble CD163 levels >10 000 ng/mL should be considered the possibility of HLH.

Adolescent ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; immunology ; Female ; Humans ; Infant ; Male ; Receptors, Cell Surface ; analysis

Acute monocytic leukemia is a distinct subtype of acute myeloid leukemia (AML) with characteristic biology and clinical features. This study was designed to compare the immunophenotypical features and clinical manifestations of the patients with AML-M(5a) to that of patients with AML-M(5b), and to identify differences between M(5a) and M(5b) and to explore their relations. A total of 58 cases of de novo adult patients with AML M(5) were investigated. Immunofluorescence analysis by flow cytometry was performed to determine the immunophenotype of the leukemic cells in all cases. Meanwhile, clinical data of these cases were studied retrospectively. The results showed that the immunophenotypes of monocytic leukemic cells in patients with AML M(5) were heterogeneous, and CD68 and CD11b were expressed higher in patients with AML M(5a), compared with that in patients with AML M(5b) (P < 0.01). The significant differences in sex, extramedullary infiltration, WBC counts of peripheral blood, complete remission rate and disease-free survival (DFS > 300 days) between the patients with AML M(5a) and M(5b) did not exist (P > 0.05). It is concluded that the special individual immunophenotype features can be detected in patients with either of AML M(5a) or M(5b), and that expressions of CD68 and CD11b were much higher in M(5a). It seems that the complete remission rate and disease-free survival of patients with M(5a) and M(5b) are not different from that of currently available therapy.
Adolescent ; Adult ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; CD11b Antigen ; analysis ; Female ; Fluorescent Antibody Technique ; Humans ; Immunophenotyping ; Leukemia, Monocytic, Acute ; classification ; immunology ; Male ; Middle Aged ; Prognosis

The object of this study was to explore the effects of Panax notoginosides (PNS) on proliferation and differentiation of human CD34(+) stem/progenitor cells. CD34(+) cells were isolated from human bone marrow by using immune beads of Dynal M- 450 system. The cells were exposed to PNS at different concentrations in both liquid and semi-solid culture for 14 days. The cells were marked with monoclonal antibodies and analyzed by flow cytometry after culture. The CFU-Mix colony formation from CD34(+) cells was assayed. The results showed that: (1) The yield of CD34(+) cells after being selected by immune beads were (1.03 +/- 0.74)% out of bone marrow nuclear cells with purity of 86% - 93%. (2) PNS (10 - 25 mg/L) stimulated the proliferation of CD34(+) cells, and raised the colony numbers of CFU-Mix obviously in vitro. PNS 25 mg/L was the optimal concentration to promote proliferation of CD34(+) cells, the increasing rate of CFU-Mix colony was (34.7 +/- 16.0)%. (3) The differentiation of CD34(+) cells was induced by exposure to PNS (25, 50 and 100 mg/L) in liquid culture for 14 days. The percentages of CD33(+) and CD15(+) cells were increased after PNS exposure, which were significantly higher than those of control (P < 0.01), however CD71(+) and G-A(+) cells were no obviously difference after PNS treatment. In conclusion, Panax notoginosides not only promote the proliferation of CD34(+) cells, but also induce the differentiation committed to granulocytes.
Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Ginsenosides ; pharmacology ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Lewis X Antigen ; analysis ; Sialic Acid Binding Ig-like Lectin 3

The phenotypes of the bone marrow cells in various subtypes of myelodysplastic syndromes (MDS) and its clinical implication were explored. The antigen expression of a panel of antigens expressed in marrow cells from 30 patients with subtypes of MDS was assayed by alkaline phosphatase anti-alkaline phosphatase method. The results showed that the expression of myeloid antigens appeared abnormality, CD13 and CD33, found on granulocyte and macrophage precursors, increased, and CD15 decreased. There were no significant changes for monocytic antigen CD14 and lymphoid antigens CD7 and CD10. CD34 was increased in RAEB/RAEB-t and was not increased in RA/RAS patients. CD71, expressed by erythroblast and proliferative cells, was higher in all subtypes of MDS than that in control group. It is suggested that the bone marrow cells from MDS patients showed abnormality of more than two series of immunophenotypes, detection of immunophenotype in MDS cells might be contributed to the diagnosis and predicting prognosis.
Adult ; Aged ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Antigens, CD7 ; analysis ; Antigens, Differentiation, B-Lymphocyte ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Bone Marrow Cells ; immunology ; CD13 Antigens ; analysis ; Female ; Humans ; Immunophenotyping ; Lewis X Antigen ; analysis ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology ; Neprilysin ; analysis ; Receptors, Transferrin ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVEGene therapy of leukemia is a new and effective method. It is known to all that the pathogenesis and development of leukemia are related to a variety of genes. Survivin is a member of inhibitors of apoptotic proteins (IAP). Its cDNA was cloned from target cell protease receptor-1 (EPR-1). It is expressed in common tumors, but there is no expression in normal and mature tissues. High expression of survivin was detected in leukemic cells. The present study was conducted to examine the role of survivin in the differentiation of leukemic cells by using antisense-oligonucleotides.

METHODSHuman leukemic cell K562 was used as the model for the study. K562 cells were divided into 4 groups randomly: antisense oligonucleuotide (ASON) group, nonsense oligonucleotide (NSON) group, lipofectin group and control group. There were 5 samples in each group, and the experiment was repeated for three times. ASON was designed with the reference to targeting survivin mRNA. K562 cells were cultured in RPMI1640 contained fetal cattle serum at a concentration of about 10 percent. Cell transfection was induced by lipofectin. Forty-eight hours after thansfection, the morphology and ultrastructure were observed. Twenty-four hours and 48 hours after thansfection, the function of K562 cells was detected by benzidine staining, POX staining and NBT staining, respectively. The mean fluorescence intensity of CD33 was determined by flow cytometry. The method of immunohistochemistry was used to examine the protein level of survivin.

RESULTSAfter thansfection with ASON, the size of K562 cells was reduced, but the cytoplasm was increased. The metarubricyte, segment granulocyte, apoptotic cells could be found. Morphologically and ultrastructurally, erythroid and myelocytic differentiation was observed. The positive level of benzidine staining in ASON group (11.90 +/- 2.30 at 24 h and 18.20 +/- 2.93 at 48 h) was higher than that of NSON group, lipofectin group and control group, respectively. The positive level of POX staining in ASON group (17.40 +/- 3.54 at 24 h and 29.40 +/- 3.70 at 48 h) was also higher than that of any other groups. The positive level of NBT staining in ASON group (7.50 +/- 2.26 at 24 h and 12.10 +/- 2.63 at 48 h) was significantly higher than that of NSON group, lipofectin group and control group, respectively (P < 0.01). In ASON group, the mean fluorescence intensity of CD33 (21.43 +/- 1.61 at 24 h and 14.86 +/- 1.20 at 48 h) was significantly lower than that of any other groups (P < 0.01). After thansfection for 24 h, the protein level of survivin in ASON group was decreased significantly compared to that of control group. There was no difference in survivin protein level amongst ASON group, NSON group and lipofectin group at 24 h (P > 0.05). Forty-eight hours after thansfection, the protein level of survivin was decreased significantly.

CONCLUSIONSASON targeting survivin can induce K562 to erythroid and myelocytic differentiation. Survivin is related to differentiation of K562 cells, and it can be a target of gene therapy for leukemia.

Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Cell Differentiation ; Humans ; Inhibitor of Apoptosis Proteins ; K562 Cells ; Microtubule-Associated Proteins ; analysis ; antagonists & inhibitors ; genetics ; physiology ; Oligonucleotides, Antisense ; genetics ; Sialic Acid Binding Ig-like Lectin 3 ; Transfection

OBJECTIVEMorphologic findings of pulmonary Langerhans' cell histiocytosis were analyzed in order to delineate diagnostic features.

METHODSH&E staining and immunohistochemical studies were performed on 7 cases of pulmonary Langerhans' cell histiocytosis.

RESULTSInfiltration by Langerhan's cells was obvious in all 7 cases. Inflammatory cell infiltrates, interstitial fibrosis and focal necrosis may also be seen. The cells expressed S-100 (7/7), CD68 (3/7), and CD1a (5/5).

CONCLUSIONSIn case there is radiologic suspicion of Langerhans' cell histiocytosis, pulmonary biopsy is strongly advised for a definitive diagnosis. S-100 and CD1a immunostaining is also helpful in this respect.

Adolescent ; Adult ; Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Child ; Diagnosis, Differential ; Female ; Histiocytosis, Langerhans-Cell ; diagnosis ; metabolism ; pathology ; Humans ; Langerhans Cells ; chemistry ; Lung ; pathology ; Male ; S100 Proteins ; analysis

OBJECTIVEIt has been proposed that aberrant immunity of local bowel mucosa may cause ulcerative colitis (UC) and the tumor necrosis factor-alpha (TNF-alpha) and nuclear factor-kappa B (NF-kappa B) may play a role in the development of this disease. To investigate the role of TNF-alpha and NF-kappa B in childhood UC, the expression of TNF-alpha and NF-kappa B in the bowel mucosa and their relationship were studied.

METHODSUsing anti-CD68, anti-TNF-alpha and anti-NF-kappa Bp65 antibodies, the cytokine immunoreactivities in the bowel mucosa of 39 cases of childhood UC (active UC: n = 21, non-active UC: n = 18) were detected by immunohistochemistry. The control specimens of normal bowel mucosa were collected from 7 cases with colorectal polyp or abdominal pain by sigmoidoscopy.

RESULTSThe numbers (median: interquartile range) of CD68(+) cells, TNF-alpha(+) cells and NF-kappa Bp65(+) cells were 44.0 (31.5 - 48.2), 42.7 (19.5 - 65.0) and 50.7 (30.0 - 58.0) in the active UC mucosa, and were 9.2 (7.9 - 16.6), 5.5 (2.5 - 9.1) and 4.2 (3.0 - 8.4) in non-active UC mucosa, and 5.3 (4.3 - 8.7), 3.0 (0.0 - 6.3) and 3.3 (0.0 - 4.0) in the control mucosa, respectively. The levels of CD68, TNF-alpha and NF-kappa Bp65 expressions in the active UC were significantly higher than those in the non-active UC (P < 0.001) and the controls (P < 0.001). The expression level of CD68 in non-active UC was much higher than that in the controls (P = 0.008). Using the correlation analysis, a positive correlation between TNF-alpha and NF-kappa B activation was found (r = 0.885, P < 0.001).

CONCLUSIONSMacrophages TNF-alpha and NF-kappa B may play an important role in the pathophysiologic mechanism of childhood active UC. The activation of NF-kappa B may be associated with the release of TNF-alpha.

Adolescent ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Child ; Child, Preschool ; Colitis, Ulcerative ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Infant ; Male ; NF-kappa B ; analysis ; Tumor Necrosis Factor-alpha ; analysis