OBJECTIVETo explore the effects of oxymatrine on serum levels of Th1/Th2 cytokines in HBsAg transgenic mice.

METHODSHBsAg transgenic mice were divided into oxymatrine group and control group. Each mouse was injected with either oxymatrine 200 mg/kg 0.2 ml or 0.9% NaCl 0.2 ml intraperitoneally once a day for 30 days. Serum IFN-gamma, IL-2 and IL-4, IL-10 were quantitated before and after different treatment.

RESULTSThere was no significant difference on the levels of IFN-gamma and IL-4 before and after treatment in control group. While in oxymatrine group, the levels of IFN-gamma before and after treatment were (3.108+/-3.172) pg/ml and (11.059+/-6.971) pg/ml; those of IL-4 were (29.045+/-13.235) pg/ml and (13.024+/-9.002) pg/ml (both P less than 0.001). After treatment, the levels of IL-2 in control and oxymatrine group were (1.070+/-0.447) pg/ml and (5.537+/-2.887) pg/ml (P less than 0.000 1); and those of IL-10 were (97.226+/-73.306) pg/ml and (33.607+/-23.154) pg/ml (P less than 0.01).

CONCLUSIONAfter injection of oxymatrine to HBsAg transgenic mice, the serum concentration of Th1 cytokines increased while the Th2 cytokines decreased. This can help us understand more better on the mechanisms of anti-HBV effect of oxymatrine.

Alkaloids ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; Cytokines ; blood ; Female ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Quinolizines

OBJECTIVETo study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat.

METHODSThirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test.

RESULTSWound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05].

CONCLUSIONSMacrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.

Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, Myelomonocytic ; genetics ; metabolism ; Collagen ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukin-13 ; Interleukin-4 ; Macrophages ; Male ; Phenotype ; Rats ; Skin ; injuries ; Tumor Necrosis Factor-alpha ; blood ; Wound Healing ; genetics

Cytokines such as interleukin 10 (IL10) may play an important role in the process of inflammation. The aim of this study was to analyze the association between IL10, IL10RA and IL10RB single nucleotide polymorphisms (SNPs), and benign prostate hyperplasia (BPH) in Korean population. All patients with BPH were divided into two groups according to international porostate symptom score (IPSS), prostate specific antigen (PSA) level, Qmax, and prostate volume. We selected two IL10 SNPs (rs1518111 and rs1554286), three IL10RA SNPs (rs2256111, rs4252243, and rs2228054), and two IL10RB SNPs (rs999788 and rs2834167). Genotypes of seven SNPs were determined through direct sequencing. The G/G genotype of IL10RB polymorphism (rs2834167) was associated with a high PSA level compared with the A/G + A/A genotypes (P = 0.009). Of IL10 SNP, the A/A genotype of rs1518111 and T/T genotype of rs1554286 were associated with small prostate volume, respectively (P = 0.011, P = 0.014). Moreover, the T/T genotype of IL10RB polymorphism (rs999788) was associated with high prostatic volume compared with the T/C + C/C genotypes (P = 0.033). The linkage disequilibrium (LD) blocks were formed in IL10 and IL10RA. However, haplotypes in the LD block were not associated with BPH. It is concluded that there is a strong association between the IL10 and IL10RB SNPs, and BPH in Korean population.
Aged ; *Genetic Predisposition to Disease ; Genotype ; Humans ; Inflammation ; Interleukin-10/*genetics ; Interleukin-10 Receptor alpha Subunit/*genetics ; Interleukin-10 Receptor beta Subunit/*genetics ; Linkage Disequilibrium ; Male ; Middle Aged ; *Polymorphism, Single Nucleotide ; Prostate-Specific Antigen/blood/genetics ; Prostatic Hyperplasia/*genetics ; Republic of Korea ; Sequence Analysis, DNA

OBJECTIVETo investigate the effect of IL-10 and the methylation of its promoter in acute on chronic liver failure (ACLF).

METHODSPatients were divided into three groups: 25 with ACLF, 25 with CHB, 10 healthy controls. Respectively detect the serum level of IL-10 via ELISA, and the methylation of IL-10 promoter via MSP, to analyze the difference among the three groups.

RESULTSBoth the ACLF group and the CHB group have significant increase in serum level of IL-10 compared with the control group (P < 0.05); the ACLF group's level is higher than the CHB group, however without statistical significance (P > 0.05). The serum level of IL-10 in ACLF group has no significant relativity with ALT and HBV-DNA( r = -0.022, r = 0.033, respectively; P > 0.05); has positive relativity with TBIL and MELD ( r = 0.566, r = 0.443, respectively; P < 0.05); and negative relativity with PTA (r = -0.581, P < 0.05). The distribution of the methylation of IL-10 promoter in ACLF group is significantly different from the other two.

CONCLUSIONThe serum level of IL-10 in hepatitis patients is significantly higher and increases with the degree of liver failure. The promoter methylation may be important in the gene inactivation.

Adolescent ; Adult ; Chronic Disease ; DNA Methylation ; Female ; Humans ; Interleukin-10 ; blood ; genetics ; metabolism ; Liver Failure, Acute ; blood ; genetics ; metabolism ; Male ; Methylation ; Middle Aged ; Promoter Regions, Genetic ; Young Adult

OBJECTIVETo investigate the relationship of interleukin-10 gene (IL-10) polymorphism and the serum IL-10 level with restenosis after percutaneous coronary intervention (PCI) in Tianjin Chinese Han population and study the effect of IL-10 gene polymorphism on serum IL-10 level.

METHODSFour hundred and thirty-seven patients who successfully underwent PCI with a follow-up angiography were divided into a restenosis group (n = 166) and non-restenosis group (n = 271). The IL-10 gene promoter polymorphism at position -592 was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Meanwhile their serum IL-10 level before and 24 h after PCI was determined by enzyme-linked immunosorbent assay (ELISA).

RESULTS(1) There was no significant difference in frequencies of -592 genotypes and alleles between the two groups (P > 0.05); (2) The 24 h post-PCI IL-10 serum level of restenosis group was significantly lower than that of the non-restenosis group [(82.67 ± 35.02) ng/L vs. (95.08 ± 32.26) ng/L, P < 0.05]; (3) The serum level of the A allele carriers (AA+AC) was significantly lower than that of the CC carriers [(86.13 ± 34.77) ng/L vs. (102.50 ± 27.52) ng/L, P < 0.05]; (4) In the restenosis group, the 24 h post-PCI serum level of IL-10 in the A allele carriers was also significantly lower than that in those without the A allele [(78.51 ± 34.09) ng/L vs. (102.19 ± 33.66) ng/L, P < 0.05]; (5) Logistic regression analysis revealed positive correlations between acute coronary syndrome patients, pre-PCI degree of stenosis, length of target stenosis lesion and restenosis (OR = 5.90, 1.86, 2.83 respectively); and there were negative correlations between 24 h post-PCI serum level of IL-10, the stent diameter, the diameter of reference vessel before stent implantation and restenosis(OR = 0.99, 0.70, 0.46 respectively).

CONCLUSION(1) The IL-10 gene -592 C/A polymorphism was not associated with restenosis in the Tianjin Chinese Han population; (2) IL-10 is an early post-PCI inflammatory cytokine, 24 h post-PCI serum IL-10 level was an independent predictive factor for restenosis, the IL-10 A allele carriers may have increased incidence of in-stent restenosis (ISR) by reducing the serum IL-10 levels.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; Asian Continental Ancestry Group ; genetics ; Coronary Restenosis ; genetics ; Female ; Genotype ; Humans ; Interleukin-10 ; blood ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Stents

OBJECTIVETo study the relationship between polymorphisms of interleukin 10 (IL10QX) promoter and serum levels of lipoprotein in the healthy Chinese Han population.

METHODSPCR restriction fragment length polymorphism assay was used to detect the distribution of genotypes of IL10 -592,-819,-1082 in 200 healthy Chinese Han subjects. Serum levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and very low-density lipoprotein (VLDL) in all subjects were measured to analyze the relationship with the polymorphisms of IL10 promoter.

RESULTSComparing with AA genotype, the group with GA genotype at IL10 promoter -1082 position had a significant elevation of serum HDL-C level [(1.514+/-0.501) mmol/L vs. (1.261+/-0.346) mmol/L, t=-2.225, P=0.028] and a lower serum TG level[(1.701+/-1.836) mmol/L vs. (0.981+/-0.314) mmol/L,Z=-2.096,P=0.036]. The TG, TC, HDL-C, LDL-C and VLDL levels did not show any statistically significant differences among different genotypes (CC, AA, CA) of the IL10 -592, as well as the genotypes (TT, TA, AA) ofIL10 -819 (P>0.05).

CONCLUSIONThe results suggest that in the Chinese Han population, the polymorphism at position -1082 in the promoter region of IL10 gene may be associated with the serum HDL-C level and TG level.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Genotype ; Humans ; Interleukin-10 ; genetics ; Lipoproteins ; blood ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics ; Promoter Regions, Genetic ; genetics ; Triglycerides ; blood ; Young Adult

BACKGROUNDWhile the incidence of paroxysmal nocturnal hemoglobinuria (PNH) is relatively high in Northern China, the exact mechanism of the disease remains unknown. Immunoregulatory cytokine polymorphisms can directly regulate the expression levels of cytokines, which play a crucial role in many diseases. The purpose of this study was to study cytokine gene single nucleotide polymorphisms (SNPs) and the correlated cytokine expression levels in relationship to the PNH pathogenesis.

METHODSPeripheral blood samples were collected from 30 PNH patients and 40 healthy donors; all of the samples were collected from the Han people of Northern China. Eight SNP loci in five cytokine genes, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β), interleukin-6 (IL-6), and IL-10, and aplastic anemia (AA) were assessed. TNF-a, TGF-b, IFN-g, IL-6, and IL-10 were analyzed by sequence-specific primer polymerase chain reaction (PCR-SSP). The plasma protein levels of TNF-a, TGF-b, and IFN-g were assessed by an ELISA.

RESULTSThe PNH patients had a lower frequency of the TC/GG genotype of the TGF-b gene (P < 0.01) and a higher frequency of the C allele in the TGF-b gene (+10) compared to the controls (P < 0.05). The predominant genotype of the +874 locus of the IFN-g gene was TA in the PNH patients, while that in the predominant genotype was AA in the control group and was statistically significant (P < 0.001). The frequency of the T allele in the IFN-g gene was dramatically higher in the PNH patients than in the controls (P < 0.05). The PNH patients had a reduced frequency of the GC and CC genotypes, as well as the C allele at locus -174 of the IL-6 gene compared to the controls (P < 0.01). In addition, the plasma concentrations of TNF-a, TGF-b, and IFN-g were significantly higher in the PNH group compared to the control group (P < 0.01).

CONCLUSIONSExpression levels of the TNF-a, TGF-b, and IFN-g cytokines play an important role in PNH. The GC and CC genotypes, as well as the C allele of the IL-6 gene may protect the Han people of Northern China against PNH. Additionally, the TC/GG genotype of the TGF-b gene may be the protective allele. In contrast, the TA genotype and the T allele for the IFN-g gene, as well as the C allele of TGF-b may be susceptible to PNH. However, SNPs in the TNF-a and IL-10 genes did not correlate with PNH development. Alternatively, the increased plasma concentrations of TNF-a, TGF-b, and IFN-g in PNH patients may also be related to PNH development.

Adult ; Aged ; Alleles ; Anemia, Aplastic ; genetics ; Asian Continental Ancestry Group ; China ; Cytokines ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hemoglobinuria, Paroxysmal ; blood ; genetics ; Humans ; Interferon-gamma ; blood ; genetics ; Interleukin-10 ; blood ; genetics ; Interleukin-6 ; blood ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Transforming Growth Factor beta ; genetics ; Tumor Necrosis Factor-alpha ; blood ; genetics ; Young Adult

OBJECTIVETo investigated the association between interleukin-10(IL-10)-1082/-819/-592 promoter polymorphism, plasma IL-10 levels and risk of vocal leukoplakia.

METHODSA case-control study of 61 patients with vocal cords leukoplakia and 119 healthy subjects were performed. Genotypes for the IL-10 gene (IL-10 -1082 G/A, -819 C/T and -592 C/A) were determined using pyrosequencing and plasma IL-10 levels were analyzed by ELISA.

RESULTSSmoking and alcohol consumption increased the risk of vocal leukoplakia (OR = 4.73, 95%CI:2.4-9.1;OR = 5.70, 95%CI:2.9-11.2, P < 0.01) . Patients with vocal cord leukoplakia had significantly higher frequencies of AC at -592 and -819 (OR = 1.93, 95%CI:0.97-3.81, P = 0.05) and AG at -1082(OR = 2.14, 95%CI:0.87-5.27, P = 0.092) than control. Vocal leukoplakia patients had higher concentration of plasma IL-10 (21.6 ± 0.5) pg/ml (X(-) ± s) than controls (19.0 ± 1.1)pg/ml(t = 2.08, P = 0.043) and the haplotype containing the G allele was associated with higher plasma IL-10 concentrations (24.3 ± 5.7) pg/ml compared with the ATA haplotype(19.9 ± 4.7) pg/ml (t = -2.64, P = 0.008).

CONCLUSIONSIL-10-1082/-819/-592 promoter polymorphism and high concentration of plasma IL-10 are associated with vocal cord leukoplakia, and the concentration of plasma IL-10 is associated with the genotypes of IL-10 promoter polymorphism.

Adult ; Aged ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Humans ; Interleukin-10 ; blood ; genetics ; Leukoplakia ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Vocal Cords ; pathology

OBJECTIVETo investigate the effect of IL-10 on the inducibility of human beta-defensin 2 (hBD-2) in human peripheral blood cells.

METHODSPeripheral blood samples were collected from 22 healthy individuals and co-cultured with 0 ng/ml lipopolysaccharide (LPS), 100 ng/ml LPS, 10 ng/ml IL-10, or 100 ng/ml LPS plus 10 ng/ml IL-10 at 37 degree for 6 h. Total RNA was extracted from peripheral blood cells and the mRNA level of hBD-2 was detected by relative quantitative real-time PCR..

RESULTNo detectable level of hBD-2 mRNA was found in normal peripheral blood cells with stimulation of 0 ng/ml LPS or 10 ng/ml IL-10. The mRNA level of hBD-2 was increased to 166.9 +/- 35.14 after 100 ng/ml LPS challenge, while the mRNA level of hBD-2 was decreased to 30.40 +/- 9.18 after the co-stimulation of 100 ng/ml LPS plus 10 ng/ml IL-10; which was significantly lower than that with LPS alone (P<0.05).

CONCLUSIONThe expression of hBD-2 gene can be induced by LPS.IL-10 inhibits the inducible expression of hBD-2.

Blood Cells ; cytology ; drug effects ; metabolism ; Cells, Cultured ; Humans ; Interleukin-10 ; pharmacology ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; beta-Defensins ; genetics

Adult ; Aged ; Arteriosclerosis ; epidemiology ; prevention & control ; Female ; Genotype ; Humans ; Incidence ; Inflammation ; epidemiology ; prevention & control ; Interleukin-10 ; blood ; genetics ; Kidney Failure, Chronic ; complications ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic