OBJECTIVE: To explore the expression of CD40/CD40L in multiple myeloma(MM) patients and its influence on prognosis. METHODS: Thirty patients with MM treated in Cangzhou People's Hospital from May 2016 to June 2017 were selected and divided into MM group, then 30 healthy people with a physical examination in our hospital at the same time were selected as the normal group. The serum CD40/CD40L levels of the patients in the two groups was detected by flow cytometry, and its correlation with the lymphocyte population, pathological grade and prognostic significance of MM patients was anaysis. RESULTS: The expression of CD40 in serum of the patients in MM group was significantly higher than those in normal group (P<0.05). The expression of CD40L in serum of the patients in MM group showed no significant difference as compared with those in normal group (P>0.05). The levels of CD40 and CD40L in the patients before and after chemotherapy showed no difference(P>0.05). The levels of Ts and NK cells in the patients of MM group were lower than those in normal group (P<0.05). The proportion of total B lymphocytes, Th and Th/Ts cells between the two groups showed no significant difference (P>0.05). The CD40 level was correlated with the serum total B lymphocyte level of the patients in MM group (r=0.877, P=0.005). There was a correlation with CD40L and Th cells in the serum of MM patients (r=-0.783, P=0.035). The expression of serum CD40 in the patients at phase III-IV was higher than those of the patients at phase I-II, the levels of serum CD40L in MM patients at different periods showed no significant difference(P>0.05). The survival rate of MM patients with high CD40 expression was lower than that of MM patients with low CD40 expression (χ CONCLUSION The increasing of CD40 level in MM patients is related to the pathological grade of the patients. Chemotherapy can reduce the level of CD40. The increasing of CD40 is an important factor for the poor prognosis of MM patients. CD40L level is not meaningful for MM treatment and prognosis.
B-Lymphocytes ; CD40 Antigens ; CD40 Ligand ; Humans ; Lymphocyte Subsets ; Prognosis

OBJECTIVETo investigate the association of SNP of CD40 gene and its serum levels with ischemic stroke (IS).

METHODSA total of 202 IS patients from a hospital of Baise city were enrolled in case group from May 2013 to November 2014. At the same time, 109 healthy people who had physical check-ups in the outpatient department at the same hospital were enrolled in the control group. All participants were from Guangxi Zhuang Autonomous Region and unrelated to each other. 3 ml venous blood were collected on the premise of informed consent. The single nucleotide polymorphisms of CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A were analyzed using a Snapshot SNP genotyping assays, and the serum levels of CD40 were tested by ELISA. t-test was used to compare the serum levels of CD40 between the case and control group, and the genotypes at different locuses in case group; χ(2) test was used to compare the distribution differences of the CD40 gene locuses in different genotypes and allele between the case group and the control group; alleles was established as independent variables, the occurrence of the IS as dependent variable, and expressed relative risk with OR (95%CI) value.

RESULTSIn the case group, the frequency of CC, CT and TT genotypes in CD40 gene rs1883832 C/T were 21.78% (44/202), 49.51% (100/202) and 28.71% (58/202), respectively, and 33.17% (66/199), 48.74% (97/199), 18.09% (36/199) in the control group, respectively, the differences between the two groups was significant (χ(2)=9.57, P=0.008). The CD40 serum levels were (62.7 ± 24.5) pg/ml in the case group, which was higher than that in the control group (45.3 ± 17.2) pg/ml (t=8.97, P<0.001). The serum levels of TT and CT genotypes in CD40 gene were (65.9 ± 26.3) and (64.3 ± 25.9) pg/ml, respectively, and the differences were significant when comparing with CC genotype (t equaled 5.34 and 5.03, respectively, P<0.001). The risk of developing IS was 1.56 times higher in 1883832 T allele carriers than that in rs1883832 C allele carriers (OR=1.56, 95% CI: 1.18-2.06); Combined genotype analysis displayed that CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A polymorphisms showed strong linkage disequilibrium, the case group TCCA haplotype was tested to be associated with a significantly increased risk of IS as compared with that in the control group(OR=2.49; 95%CI: 1.13-5.48).

CONCLUSIONCD40 gene rs1883832 C/T polymorphism and its TCCA haplotype were possibly associated with ischemic stroke, and the susceptibility gene for ischemic stroke may be rs1883832 T allele.

Alleles ; CD40 Antigens ; blood ; genetics ; Case-Control Studies ; Cell Differentiation ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Polymorphism, Single Nucleotide ; Stroke ; blood ; genetics

OBJECTIVETo investigate the therapeutic effects of Qingre Quyu Granule (QQG) on the patients with severe carotid stenosis, and to explore the mechanism of it.

METHODSNinety-six patients with severe carotid stenosis were enrolled in the study and were classified into a QQG group (n=48) and a control group (n=48) randomly using consecutively numbered envelopes. The patients in the QQG group were given QQG and Western medicine, those in the control group were given Western medicine merely, the course of treatment was 16 weeks. All patients went through endarterectomy after treatment. Plaques were subjected to the analysis of CD3, CD68, soluble intercellular adhesion molecule 1 (ICAM-1), matrix metalloprotease-9 (MMP-9), CD40L, tenascin-C, and collagen content lipid content by immunohistochemistry or polarized light analysis.

RESULTSBy the end of experiment, the expressions of CD3, CD68, ICAM-1, MMP9, CD40L and tenascin-C on the plaques were statistically significant lower in the QQG group compared with the control group(P<0.01). The lipid content of the plaque was also significantly lower in the QQG group compared with the control group (P<0.01). The interstitial collagen in the tissue sections of the plaques was also significantly higher in the QQG group in comparison with the control group (P<0.01).

CONCLUSIONQQG could stabilize carotid artery plaques through inhibiting pro-inflammation factors and restraining the tenascin-C and MMP9 pathway.

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; CD3 Complex ; metabolism ; CD40 Ligand ; metabolism ; Carotid Arteries ; metabolism ; pathology ; Carotid Stenosis ; blood ; complications ; drug therapy ; Collagen ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Immunohistochemistry ; Inflammation ; complications ; pathology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipids ; blood ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Plaque, Atherosclerotic ; blood ; complications ; drug therapy ; Tenascin ; metabolism

OBJECTIVETo investigate the role of glycogen synthase kinase 3β (GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs).

METHODSMature DCs (mDCs) induced by LPS were examined for GSK-3β phosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3β in maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR; the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3β and RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3β gene.

RESULTSLPS exposure significantly lowered GSK-3β activity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3β obviously down-regulated the expression of RelB.

CONCLUSIONSGSK-3β is a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.

Animals ; B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cell Differentiation ; Cells, Cultured ; Culture Media ; chemistry ; Dendritic Cells ; enzymology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Indoles ; chemistry ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Lentivirus ; Lymphocyte Culture Test, Mixed ; Maleimides ; chemistry ; Mice ; Myeloid Cells ; enzymology ; Phosphorylation ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Signal Transduction

OBJECTIVETo study whether co-stimulatory molecule CD40 of alveolar macrophage (AM) participated in the occurrence and development of silicosis, and to explore the intervention of Yiqi Huoxue Decoction (YHD) in the fibrosis of silicosis patients.

METHODSTotally 46 silicosis inpatients and outpatients were recruited and randomly assigned to the Western treatment group (A) and the Chinese medicine (CM) treatment group (B), 23 in each group. Patients in Group A received routine symptomatic treatment such as anti-inflammation, phlegm resolving, anti-spasm, and asthma relief, and so on. Patients in Group B additionally took YHD, one dose daily for 14 successive days. Besides, another 18 patients with chronic cough and sense of laryngeal foreign bodies were recruited as the normal control group, who had no obvious lesion confirmed by bronchofi6roscope and clinical diagnosis of the lung. They were treated by symptomatic supporting treatment. The alveolar lavage fluid was collected from all patients and isolated, and AM cells were cultured. The level of CD40 mRNA was detected by RT-PCR. The expression of CD40 protein was detected by Western blot.

RESULTSCompared with the normal control group, expression levels of CD40 mRNA and CD40 protein significantly increased in Group A (P < 0.01). Compared with Group A, expression levels of CD40 mRNA and CD40 protein significantly decreased in Group B (P < 0.01).

CONCLUSIONSHighly expressed co-stimulatory molecule CD40 of AM might participate in pulmonary fibrosis. YHD could hinder its roles, inhibit the progression of fibrosis, thereby playing an interventional role of treatment.

CD40 Antigens ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Fibrosis ; Humans ; Lung ; Male ; Pulmonary Fibrosis ; RNA, Messenger ; metabolism ; Silicosis ; drug therapy ; metabolism

OBJECTIVETo study the frequency distribution of single nucleotide polymorphisms (SNPs) in two genes associated with incomplete Kawasaki disease (KD) (rs1569723 in CD40 gene and rs2736340 in BLK gene), and to investigate its association with the genetic susceptibility and clinical phenotypes of incomplete KD.

METHODSA total of 184 children with incomplete KD and 203 normal children were recruited to carry out a case-control study. The genotypes of SNPs in CD40 gene and BLK gene were determined using polymerase chain reaction-restriction fragment length polymorphism. The frequency distribution of genotypes was compared between the KD and control groups. The association between gene polymorphisms and clinical features of incomplete KD was analyzed.

RESULTSThere were no significant differences in genotype (AA, AC, CC) and allele frequencies in CD40 SNP rs1569723 between the KD and control groups. There were significant differences in the frequency distribution of three genotypes (TT, CT, CC) in BLK SNP rs2736340 between the KD and control groups (P=0.031), and the KD group had a significantly higher frequency of T allele than the control group (P=0.007). There were significant differences in the incidence of conjunctival hyperaemia among the patients with different genotypes (rs1569723 in CD40 gene) (P=0.036). The SNP rs2736340 in BLK gene was associated with the extremity changes in KD patients (P=0.017).

CONCLUSIONSThe SNP rs2736340 in BLK gene is associated with the susceptibility to incomplete KD, and the SNP rs1569723 in CD40 gene and SNP rs2736340 in BLK gene are associated with some of clinical phenotypes of incomplete KD.

CD40 Antigens ; genetics ; Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymorphism, Single Nucleotide

BACKGROUND: Anti-Gal is a major antibody induced in non-human primates (NHPs) after xenotransplantation. To understand the mechanism of graft rejection, we investigated the association between anti-Gal responses and graft failure in NHP recipients of porcine islet transplantation (PITx). METHODS: Intraportal PITx was performed in 35 diabetic NHPs, and graft function was monitored. Early graft failure (EGF) was defined as loss of graft function within a month after PITx. Seven, 19, nine NHPs received immunosuppression (IS) without CD40 pathway blockade (Group I), with anti-CD154 (Group II), and with anti-CD40 (Group III), respectively. The anti-Gal levels on day 0 and day 7 of PITx were measured by ELISA. RESULTS: The frequency of EGF was significantly lower in Group II (26.3%) than in Group I (100%, P=0.0012) and Group III (77.8%, P=0.0166). While levels of anti-Gal IgG in Group I and anti-Gal IgM in Group III increased on day 7 compared with day 0 (P=0.0156 and 0.0273), there was no increase in either on day 7 in Group II. The ratio of anti-Gal IgM or IgG level on day 7 to that on day 0 (Ratio7/0) was significantly higher in recipients with EGF than without EGF (P=0.0009 and 0.0027). ROC curve analysis of anti-Gal IgM Ratio7/0 revealed an area under the curve of 0.789 (P=0.0003). CONCLUSIONS: IS with anti-CD154 suppressed anti-Gal responses and prevented EGF in PITx. Anti-Gal IgM Ratio7/0, being associated with EGF, is a predictive marker for EGF.
Animals ; Antibodies/blood/immunology ; Antigens, CD40/immunology ; Area Under Curve ; CD40 Ligand/immunology ; Disaccharides/*immunology ; Epidermal Growth Factor/blood ; Graft Rejection/*immunology ; Immunoglobulin G/blood ; Immunoglobulin M/*blood ; Immunosuppressive Agents/therapeutic use ; *Islets of Langerhans Transplantation ; Macaca mulatta ; ROC Curve ; Swine ; Transplantation, Heterologous

PURPOSE: The aim of this study was to assess the expressions of CD44 and CD133 in colorectal cancer tissue by using immunohistochemical staining and to analyze the clinical significance of the expressions related to other clinicopathological data and survival results. METHODS: One hundred sixty-two patients with a biopsy-proven colorectal adenocarcinoma who were operated on between January 1998 and August 2004 were enrolled in this study. Immunohistochemical staining for CD44 and CD133 was performed on primary colorectal cancer tissue, metastatic lymph nodes, and synchronous and metachronous metastatic tumor tissues if available. RESULTS: CD44 expression was stronger in the primary tumor than in metastatic lymph nodes (P < 0.001), and CD133 expression tended to be stronger in primary tumor than in metastatic lymph nodes (P = 0.057). No significant correlation was found between the CD44 and the CD133 expressions. The cases with recurrence showed low expression of CD44 (P = 0.017). CD133 expression was lower in cases with elevated CA 19-9 serum levels (P = 0.028) and advanced T stage (P = 0.038). Multivariate analysis proved that low expression of CD44 was an independent prognosis factor for short disease-free survival (P = 0.028). CONCLUSION: Low CD44 expression was correlated with increased tumor recurrence and short disease-free survival, and low CD133 expression was associated with advanced tumor stage. We suggest that further studies be performed to evaluate whether the immunohistochemical method for determining the CD44 and the CD133 expressions is appropriate for exploring cancer stem-cell biology in patients with colorectal cancer.
Adenocarcinoma ; Antigens, CD40 ; Biology ; Colorectal Neoplasms* ; Disease-Free Survival ; Humans ; Lymph Nodes ; Multivariate Analysis ; Neoplastic Stem Cells* ; Prognosis ; Recurrence ; Stem Cells

The aim of the study was to investigate the anti-asthmatic effects of oxymatrine (OXY) and the possible underlying mechanisms. The mouse asthma model was established by ovalbumin (OVA) intraperitoneal injection. A total of fifty mice were randomly assigned to five groups: control, OVA, OVA + dexamethasone (Dex, 2 mg · kg(-1)), and OVA + OXY (40 mg · kg(-1)), and OVA + OXY (80 mg · kg(-1)), respectively. Histological studies were conducted by the hematoxylin and eosin (HE) staining, the levels of interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13, and IgE were evaluated by enzyme-linked immunosorbent assay (ELISA), and the protein level of CD40 was analyzed by Western blotting. OXY inhibited OVA-induced increases in eosinophil count; the levels of IL-4, IL-5, IgE, and IL-13 were recovered. It also substantially inhibited OVA-induced eosinophilia in lung tissues and the expression of CD40 protein. These findings suggest that OXY may effectively ameliorate the progression of asthma and could be explored as a possible therapy for patients with allergic asthma.
Alkaloids ; pharmacology ; Animals ; Anti-Asthmatic Agents ; pharmacology ; Anti-Inflammatory Agents ; pharmacology ; Asthma ; drug therapy ; Bronchoalveolar Lavage Fluid ; chemistry ; CD40 Antigens ; metabolism ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Female ; Immunoglobulin E ; metabolism ; Interleukins ; metabolism ; Irritants ; toxicity ; Mice, Inbred BALB C ; Ovalbumin ; toxicity ; Pulmonary Eosinophilia ; chemically induced ; drug therapy ; Quinolizines ; pharmacology ; Random Allocation ; Signal Transduction ; drug effects

OBJECTIVE: To investigate the expression of inflammatory molecule CD40 in the pallium and hippocampus of rats after status epilepticus (SE).
 METHODS: The expression of CD40 in the pallium, the different areas of hippocampus and the different cells from the lithium-pilocarpine epileptic rats at different time points were examined by immunohistochemistry and double-immunofluorescent labeling.
 RESULTS: After SE, CD40 expression was obviously inhibited, especially in hippocampus. CD40 was mainly expressed in the activated microglia. CD40 positive cells reached a peak at the 3rd day and returned to a slightly higher level at the 7th day after SE compared with the level before SE.
 CONCLUSION Elevation of CD40 expression in the activated microglia can promote inflammatory injury of rat's hippocampus, suggesting that CD40 induced-signal pathway is involved in inflammatory injury in the hippocampus after SE.
Animals ; CD40 Antigens ; metabolism ; Epilepsy ; Hippocampus ; metabolism ; Immunohistochemistry ; Lithium ; Microglia ; metabolism ; Pilocarpine ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus