Objective To investigate the relationship between disease courses and severity and monocyte subsets distribution and surface CD31 intensity in patients of hemorrhagic fever with renal syndrome (HFRS). Methods Peripheral blood samples from 29 HFRS patients and 13 normal controls were collected. The dynamic changes of classical monocyte subsets (CD14⁺⁺CD16^-), intermediated monocyte subsets (CD14⁺⁺CD16⁺) and non-classical monocyte subsets (CD14⁺CD16⁺⁺) and the mean fluorescent intensity (MFI) of CD31 on monocyte subsets were detected by multiple-immunofluorescent staining and flow cytometry. Results In acute phase of HFRS, the ratio of classical monocyte subsets to total monocytes was dramatically decreased compared to convalescent phase and normal control. It was still much lower in convalescent phase compared to normal controls. The ratio of classical monocyte subsets to total monocytes were decreased in HFRS patients compared to that in normal control, whereas there was no difference between severe/critical groups and mild/moderate groups. On the contrary, the ratio of intermediate monocyte subsets to total monocytes in acute phase of HFRS was significantly increased compared to convalescent phase and normal control. The ratio of intermediate monocyte subsets to total monocytes were increased in HFRS patients compared to that in normal control, whereas no difference was found between severe/critical groups and mild/moderate groups. Phases or severity groups had no difference in ratio of non-classical monocyte subsets to total monocytes. Additionally, the ratio of classical monocyte subsets had a tendency to decline and that of intermediate monocyte subsets showed an increase both to total monocytes between the acute and convalescent phases in 11 HFRS patients with paired-samples. Moreover, in acute phase of HFRS, the mean fluorescent intensity (MFI) of CD31 on three monocyte subsets all decreased, specifically classical monocyte subsets showed the highest MFI of CD31 while the normal control reported the highest MFI of CD31 in non-classical monocyte subsets. In convalescent phase, the MFI of CD31 on classical and intermediated monocyte subsets were both lower than that of normal control, while MFI of CD31 was still significantly lower than normal control on non-classical monocyte subsets. Finally, MFI of CD31 on classical and intermediated monocyte subsets in severe/critical group were both lower than those in mild/moderate group, showing no statistical difference in MFI of CD31 on non-classical monocyte subset across groups of different disease severity. Conclusion The ratio of classical and intermediated monocyte subsets to total monocytes are correlated with the course of HFRS, and so are the surface intensity of CD31 on these monocyte subsets with the disease course and severity. The surface intensity of CD31 on non-classical monocyte subsets, however, is correlated only with the course of the disease. Together, the underlying mechanisms for the observed changes in monocyte subsets in HFRS patients should be further investigated.
Humans ; Monocytes ; Lipopolysaccharide Receptors ; Hemorrhagic Fever with Renal Syndrome ; Receptors, IgG ; Disease Progression

OBJECTIVETo study the clinical and laboratory characteristics of juvenile myelomonocytic leukemia (JMML).

METHODSThe clinical characteristics and laboratory results were retrospectively analyzed in 10 children with newly diagnosed JMML. They were compared with those of 28 children with myelodysplastic syndrome (MDS) and 44 children with chronic myeloid leukemia (CML).

RESULTSCompared with the children with CML or MDS, the children with JMML had significantly higher rates of skin rashes, ecchymosis, and lymphadenectasis, a significantly lower serum cholinesterase (ChE) level, and a significantly higher fetal hemoglobin level (P<0.05). The white blood cell count of children with JMML was significantly higher than that of children with MDS, but significantly lower than that of children with CML (P<0.05). In addition, the myeloid/erythroid ratio and rate of dyshaematopoiesis were significantly lower in children with JMML than those in children with CML or MDS. The children with JMML had a significantly higher expression of mature monocyte marker CD14 than those with CML or MDS (P<0.05). The levels of myeloid markers CD33, CD11b, CD13, and CD15 in children with JMML were significantly higher than those in children with MDS, but significantly lower than those in children with CML (P<0.05). The levels of CD2 and CD7 in children with JMML were higher than those in children with CML, but lower than those in children with MDS (P<0.05).

CONCLUSIONSSkin rashes, ecchymosis, lymphadenectasis, and ChE reduction are more common in children with JMML than in those with CML or MDS, while dyshaematopoiesis is less common. In addition, CD14 level increases significantly in children with JMML.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukemia, Myelomonocytic, Juvenile ; genetics ; immunology ; Lipopolysaccharide Receptors ; analysis ; Male

OBJECTIVETo establish a transgenic cell line with stable expression of CD14.

METHODSTotal RNA extracted from peripheral blood mononuclear cells was treated with RNAase-free DNAase, the human CD14 gene was cloned and sequenced through the RT-PCR, T-A clone techniques and ABI PRISM377 machine. Eukaryotic expression vector pcDNA3.1(+)/CD14 was constructed by cleaving with double restriction endonucleases EcoR I/Xba I and ligating with T4 ligase. The human cervical cancer cell line Hela was transfected with the positive recombinant plasmid pcDNA3.1(+)/CD14 using superfect transfection reagent. Positive clones were selected by G418 at a concentration of 0.5 μg/μl and the expression of human CD14 on the transfected Hela cells was confirmed by quantitative PCR and immunofluorescent assay.

RESULTSThere was significantly difference om expression of CD14 mRNA between the blank pcDNA3.1(+) transfected cells and pcDNA3.1(+)/CD14 transfected cells (P<0.01). The fluorescence was significantly stronger on the stable cell line Hela-CD14 than that on the transiently transfected Hela cells,and no visible fluorescence was observed in blank vector transfected cells.

CONCLUSIONThe transfectant cell line Hela-CD14 with stable expression of human CD14 has been successfully established, which can be used to study human CD14 molecular and CD14-associated monocyte/macrophage cell diseases.

Female ; Gene Expression ; Genetic Vectors ; HeLa Cells ; Humans ; Lipopolysaccharide Receptors ; genetics ; metabolism ; Plasmids ; genetics ; Transfection

OBJECTIVETo observe the effect of salvianolic acid B (SAB), an extract from Radix Salviae miltiorrhizae, on expression of leucocyte differentiation antigen 14 (CD14) in the liver tissue of experimental rats with carbon tetrachloride (CCl4)-induced liver fibrosis.

METHODSThirty SD rats were randomly divided into three groups, the model group, the treated group, and the control group. The pathological fibrosis changes in liver of rats were observed. Meantime, their liver function was detected by automatic biochemical analyzer. Serum content of endotoxin was assayed by matrix staining, and plasma content of tumor necrosis factor-alpha (TNF-alpha) was detected by radioimmunoassay. mRNA and protein expressions of CD14 in the liver tissue were measured using reverse transcriptional-polymerase chain reaction and immunohistochemistry respectively.

RESULTSAll the laboratory parameters, including liver function, degree of liver fibrosis, serum endotoxin levels, plasma TNF-alpha contents, and CD14 mRNA and protein expressions in the model group were higher than those in the control group (all P<0.01). All the aforesaid indices were lowered more in the treated group than in the model group (all P<0.01).

CONCLUSIONSSAB could antagonize the CCl4, induced liver fibrosis in rats. Its mechanism of action was possibly correlated with its effects on down-regulating hepatic CD14 expression and blocking the endotoxin signal transduction pathway.

Animals ; Benzofurans ; pharmacology ; Lipopolysaccharide Receptors ; metabolism ; Liver ; drug effects ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

This study aimed to examine the number of circulating Toll-like receptor 4 (TLR4) + CD14+ monocytes in patients with different stages of chronic kidney disease (CKD), their responses to lipopolysaccharide (LPS), and to explore the potential association of the number of TLR4+CD14+ monocytes with clinical laboratory measures. The numbers of TLR4+CD14+, LPS-stimulated TNF-α+CD14+ and interleukin (IL)-6+CD14+ monocytes were determined by flow cytometry in 9 patients with stage 3 CKD, 11 with stage 4 CKD, 16 with stage 5 CKD, and 19 healthy controls (HCs). Their laboratory tests were performed by routine methods and the potential association among these measures was analyzed by Pearson's correlation analysis. The numbers of CD14+, CD14+TLR4+, LPSstimulated TNF-α+CD14+ and IL-6+CD14+ monocytes in patients with CKD were significantly less than those of HCs (all P<0.05), and were negatively associated with patient disease severity. The number of CD14+TLR4+ monocytes was positively correlated with estimated glomerular filtration rate (eGFR, P<0.001) and the levels of hematocrit (P<0.01), but negatively correlated with the levels of blood urine nitrogen, serum creatinine, and C-reactive protein (P<0.001 for all), in the CKD patients. Our data indicate that significant reduction in the number of TLR4+ monocytes and their impaired responses to LPS may be associated with the progression of CKD in Chinese patients.
Cytokines ; metabolism ; Humans ; Kidney Failure, Chronic ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; pharmacology ; Monocytes ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVE: To investigate the role of myeloid-derived suppressor cells (MDSC) in the prognosis of patients with diffuse large B cell lymphoma (DLBCL). METHODS: The peripheral blood of 52 DLBCL patients and 30 healthy volunteers was collected. The CD14HLA-DR was used as the immune marker for MDSC. The role of MDSC in the prognosis of DLBCL patients was analyzed by combination with the related clinicopathological data. RESULTS: The proportion of MDSC in peripheral blood of newly diagnosed DLBCL patients increased significantly (P＜0.01). The expression of MDSC in DLBCL patients was related with clinical staging, lactate dehydrogenase (LDH) level and IPI score (P＜0.01). There was no significant correlation with sex, age, and B symptoms. Univariate analysis showed that the clinical stage, serum LDH level, IPI score and MDSC level were the adverse factors affecting the overall survival (OS). Multivariate analysis showed that IPI score and MDSC level were independent risk factors for OS in DLBCL patients. CONCLUSION MDSC can be used as an important index to evaluate the prognosis of DLBCL patients, contributing to evaluate the immune and tumor microenvironment of DLBCL patients.
Biomarkers ; HLA-DR Antigens ; Humans ; Lipopolysaccharide Receptors ; Lymphoma, Large B-Cell, Diffuse ; Myeloid-Derived Suppressor Cells ; Prognosis ; Tumor Microenvironment

OBJECTIVE: To investigate the clinical significance of tissue factor (TF) and vascular endothelial growth factor (VEGF) expression on peripheral blood CD14 positive monocytes in patients with diffuse large B cell lymphoma (DLBCL). METHODS: The expressions of TF and VEGF on peripheral CD14 monocytes in 41 patients with DLBCL (DLBCL group) before chemotherapy and after 4 chemotherapeutic courses, and in 20 healthy subjects (control group) were detected by flow cytometry respectively, meanwhile, the relationship of the expression of TF and VEGF with international prognostic indexes (IPI) and short-term effects were analysed. RESULTS: The expression levels of TF and VEGF on peripheral CD14 monocytes in DLBCL group were significantly higher than those in control group (P<0.01), and a positive correlation was found between the two groups (r=0.755, P<0.01). The expression of TF and VEGF on CD14 monocytes in patients with prognostic risk factors significantly increased as compared with those in patients without prognostic risk factors (P<0.05), but there were no significant differences of TF and VEGF expressions on CD14 monocytes in DLBCL group with different sex, age, subtypes (P＞0.05). As compared with patients without prognostic risk factors, the expression levels of TF and VEGF on CD14 monocytes of patients with prognostic risk factors significantly increased (P<0.05). The expression of TF and VEGF on CD14 monocytes in DLBCL group showed an increasing tendency along with the increase of IPI index (P<0.01). The expression levels of TF and VEGF on CD14 monocytes in remission group before chemotherapy were lower than those in non-remission group (P<0.01); after chemotherapy, the expression levels of TF and VEGF on CD14 monocytes in remission group were lower than those before chemotherapy (P<0.05), while the TF and VEGF expression levels in non-remission group were no singnificauly different from TF and VEGF levels before chemtherapy (P>0.05), the survival of patients in group with low expression of TF and VEGF was superior to that in group with high expression of TF and VEGF (P<0.05). CONCLUSION The paripheral blood CD14 monocytes in DLBCL patients highly express the TF and VEGF, which relate with IPI, therapeutic efficacy and survival, thus the TF and VEGF expression levels are of reference significance for evaluating the therapeutic efficacy and prognosis of patients.
Antineoplastic Combined Chemotherapy Protocols ; Humans ; Lipopolysaccharide Receptors ; Lymphoma, Large B-Cell, Diffuse ; Monocytes ; Prognosis ; Thromboplastin ; Vascular Endothelial Growth Factor A

Objective: To investigate the changes of intestinal wall barrier function and its correlation with infection occurrence in patients with cirrhotic portal hypertension. Methods: 263 patients with cirrhotic portal hypertension were split into: the clinically evident portal hypertension (CEPH) combined with infection group (n = 74); CEPH group (n = 104); and Non-CEPH group (n = 85). Among them, 20 CEPH patients and 12 non-CEPH patients in non-infection status were subjected to sigmoidoscopy. Immunohistochemical staining was used to detect the expression of trigger receptor-1 (TREM-1), CD68, CD14, the inducible nitric oxide synthase molecule, and Escherichia coli (E.coli) in the medullary cells of the colon mucosa. An enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of soluble myeloid cell trigger receptor-1 (sTREM-1), soluble leukocyte differentiation antigen-14 subtype (sCD14-ST) and intestinal wall permeability index enteric fatty acid binding protein (I-FABP). Fisher's exact probability method, one-way ANOVA, Kruskal-Wallis-H test, Bonferroni method, and Spearman correlation analysis were used for statistical analysis. Results: The serum sTREM-1 and I-FABP levels were higher in CEPH patients than those of non-CEPH patients in the non-infectious state (P < 0.05), but the difference in blood sCD14-ST levels was not statistically significant (P > 0.05). Serum levels of sTREM-1, sCD14-ST, and I-FABP in infected patients were higher than those in patients without a concurrent infection (P < 0.05). Serum sCD14-ST levels were positively correlated with serum sTREM-1, C-reactive protein (CRP), and procalcitonin (PCT), and sTREM-1 levels were also positively correlated with CRP and PCT (r > 0.5, P < 0.001). The rates of CD68, inducible nitric oxide synthase, CD14-positive cells, and E.coli-positive glands were higher in the intestinal mucosa of the CEPH group than those of the control group (P < 0.05). Spearman's correlation analysis showed that the rate of E.coli-positive glands in CEPH patients was positively correlated with the expression of molecular markers CD68 and CD14 in the lamina propria macrophages. Conclusion: Patients with cirrhotic portal hypertension have increased intestinal permeability and inflammatory cells, accompanied by bacterial translocation. Serum sCD14-ST and sTREM-1 can be used as indicators to predict and evaluate the occurrence of infection in patients with cirrhotic portal hypertension.
Humans ; Nitric Oxide Synthase Type II ; Lipopolysaccharide Receptors ; Prospective Studies ; Biomarkers ; C-Reactive Protein/analysis* ; Liver Cirrhosis/complications* ; Hypertension, Portal

OBJECTIVEThe expression of hepatic lipopolysaccharide (LPS) receptors in a rat nonalcoholic steatohepatitis (NASH) model was studied in order to explore the pathogenesis of NASH.

METHODSForty-five male SD rats were fed with a high fat diet. These rats were sacrificed after high fat feeding at 8, 12, 16, 24 weeks. Hepatic expressions of CD14 were observed by immunohistochemistry and expressions of TLR4 were detected by RT-PCR. Hepatic expressions and serum levels of TNFa were measured by RT-PCR and ELISA. Some rats fed with normal rat food served as controls.

RESULTSAt the 8th week fatty livers appeared, and hepatic expressions of CD14 (25.9+/-1.9) and TLR4mRNA (1.75+/-0.81) were upregulated compared to those in the control group (25.9+/-1.9 vs 12.4+/-0.7, 1.75+/-0.81 vs 0.98+/-0.33, P < 0.01, t > 2.756 and P < 0.05, t > 2.045). The hepatic expressions of the two kinds of receptors increased with the appearance of NASH at week 12 (61.8+/-1.9 and 1.88+/-0.72, P < 0.01, t > 2.756 and P < 0.05, t > 2.045), They reached to their peaks at week 16 (71.5+/-1.3 and 5.64+/-0.87, both P < 0.01 and t > 2.756), and decreased slightly at week 24 (67.7+/-6.6 and 4.98+/-0.72, both P < 0.01 and t > 2.756). Hepatic expressions and serum levels of TNFa also increased starting at week 8, and remained at that high level from week 8 to week 24.

CONCLUSIONThe hepatic expressions of CD14 and TLR4 were up-regulated gradually in the established rat NASH model. It may be one of the factors responsible for the increase of hepatic sensitivity to LPS injury of the NASH rats and may play an important role in the pathogenesis of NASH.

Animals ; Fatty Liver ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Liver ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

OBJECTIVESTo study the expression of CD34, CD31, CD14, CD10 and factor VIII on blood island of yolk sac (YS), PAS/aorta-germen-mesonephros (AGM) region and hepatic hematopoietic foci.

METHODSThirty-two cases of 3rd-12th weeks human embryo were obtained by drug abortion. Paraffin embedded sections with H.E staining and immunohistochemistry reaction (SABC) were performed.

RESULTSYS blood island of 3rd-4th weeks of gestation was consisted of two types of cells. One was vascular endothelial cells located outside and the other hematopoietic cells inside the blood island. Both the two types of cells were CD10, CD14, CD31 and factor VIII positive. Hematopoietic cells were CD34 negative, and vascular endothelial cells were CD34 positive. On 32nd days of gestation, the hematopoietic cells migrated out of YS. On 4th week of gestation, CD34, CD14, CD10, CD31 and factor VIII positive cells appeared in the aorta, mesonephros and hepatic hematopoietic foci. By the 7th week, the number of positive hematopoietic cells reached the peak. In 11th-12th weeks, most cells in these regions were matured red blood cells and were negative for all the antibodies mentioned above excepting for CD34. During 4th-12th weeks, all endothelial cells in embryo were CD34 positive.

CONCLUSIONSThe hematopoietic cells and endothelial cells of YS blood island co-expressed CD10, CD14, CD31 and factor VIII. Endothelial cells were CD34 positive but hematopoietic cells were negative in YS blood island. The hematopoietic cells of aorta, mesonephros and hepatic hematopoietic foci expressed CD34, CD10, CD14 and factor VIII from 4th week to 7th week. Anti-CD34 antibody could label endothelial cells of every kinds vessels of embryo from 3rd to 12th weeks.

Antigens, CD34 ; metabolism ; Factor VIII ; metabolism ; Fetus ; metabolism ; Humans ; In Vitro Techniques ; Lipopolysaccharide Receptors ; metabolism ; Liver ; metabolism ; Neprilysin ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Yolk Sac ; metabolism