Clinical significance of elevated serum interleukin 15 in patients with active lupus nephritis

AIM: To detect interleukin 15 (IL-15) levels in peripheral blood from patients with active lupus nephritis and investigate its clinical significance. METHODS: IL-15 level was determined by enzyme linked immunosorbent assay (ELISA). The peripheral blood mononuclear cells (PBMCs) were isolated with grads density abaxiality. The inhibitory effects of dexamethasone on production of IL-15, IgG and anti-dsDNA antibody in cultured PBMCs from LN patients were also investigated. RESULTS: (1) Serum IL-15 level in LN patients was significantly higher than that in normal controls (P<0.01). Serum IL-15 level in active LN patients was significantly higher than that in remised patients (P<0.05). (2) Serum IL-15 level was positively correlated with SLEDAI, anti-dsDNA antibody and 24 h urine protein excretion in active LN patients. (3) Serum IL-15 level was significantly reduced in LN patients treated with combination of cyclophosphamide (CTX) and steroid for 12 weeks. (4) Secretion of IL-15, IgG and dsDNA antibody in cultured PBMCs from active LN patients was significantly higher than that in normal control group, and IL-15 level in supernatant of cultured PBMCs from active LN patients was positively correlated with IgG and dsDNA antibody. Dexamethasone inhibited the secretion of IL-15, IgG and dsDNA antibody in cultured PBMCs from LN patients. CONCLUSION: Serum IL-15 level in patients with active lupus nephritis is significantly elevated, suggesting that IL-15 may be involved in the pathophysiological process of LN. IL-15 may be used as an index to assess the activity of LN.     

Correlation between Toll-like receptor 4 on peripheral blood monocytes and serum IL-6 in patients with chronic severe hepatitis B

AIM:To study the change of Toll like receptor 4(TLR4) on peripheral blood monocytes (PBMCs) and its role in the pathogenesis of chronic severe hepatitis B.METHODS:The expression of TLR4 on CD14+ PBMCs was determined by flow cytometry in 30 healthy control,31 patients with chronic hepatitis B and 30 patients with chronic severe hepatitis B. The level of serum interleukin-6 (IL-6) was detected by ELISA. RESULTS:The expressions of TLR4 on PBMCs and serum IL-6 in the groups of healthy control,patients with chronic hepatitis B and patients with chronic severe hepatitis B were 2.3±1.1,3.7±2.3,6.9±4.1 mean fluorescence intensity (MFI) and （11.5±7.2) ng/L,(40.8±31.2) ng/L,(77.6±33.3） ng/L. The TLR4 value in the group of patients with chronic severe hepatitis B was significant higher than that in the group of healthy control and the group of patients with chronic hepatitis B (P<0.05). However,there was no significant difference between the group of patients with chronic hepatitis B and the group of healthy control (P>0.05). Serum IL-6 increased gradually and significantly between the group of healthy control and the groups of patients with chronic hepatitis B and patients with chronic severe hepatitis B. There was a significant positive correlation between the expression of TLR4 and the content of serum IL-6 in the group of chronic severe hepatitis B. CONCLUSION:TLR4 may play a role in the pathogenesis of chronic severe hepatitis B.     

Preliminary study of Sonic Hedgehog signaling pathway in rheumatoid arthritis

AIM: To investigate the expression of Sonic Hedgehog (Shh) signaling pathway-associated factors in peripheral blood mononuclear cells (PBMCs) and synovial tissues of rheumatoid arthritis (RA). METHODS: The mRNA expression levels of Shh, Ptch1 and Gli1 in PBMCs of 35 RA patients, and 35 age-and sex-matched healthy controls were analyzed by real-time PCR. The expression of Shh, Ptch1 and Gli1 in synovial tissues was detected by immunohistochemisty assay in 10 RA patients and 5 patients with traumatic or meniscal injury (no arthritis) as control group. All patients accorded with the American College of Rheumatology (ACR) 1987 revised classification criteria for determining RA, and the score of DAS28 was ≥3.2. RESULTS: The results of real-time PCR showed that the expression of Shh and Gli1 mRNA in RA patients was higher than that in the controls (Shh and Gli1 in RA were 1.36±1.48 and 1.15±0.68, while Shh and Gli1 in control group were 0.47±0.25 and 0.49±0.05, respectively). The mRNA expression of Ptch1 between the 2 groups had no significant difference. Similarly, the results of immunohistochemistry assay showed that the positive staining rates of Shh and Gli1 in RA group were higher than those in control group. However, no difference of Ptch1 positive staining rate between the 2 groups was observed (P>0.05). CONCLUSION: The positive expression of Shh and Gli1 indicates the activation of Shh signaling pathway in the RA patients.     

Elevated levels of serum IL-6, ICAM-1 and P-selectin in stable survivors with liver transplantation

AIM: To study the profile of serum IL-6， ICAM-1 and P-selectin in stable survivors with clinical liver transplantation (LTx). METHODS: Flow cytometric analysis was used to determine the phenotype of T cell subsets in peripheral blood mononuclear cells （PBMCs） from stable survivors with liver transplantation (n=22), and healthy volunteers (n=12). Serum levels of the pro-inflammatory cytokines, tumor necrosis factor (TNF-α) and interleukin (IL-6), intercellular adhesion molecules (ICAM-1) and P-selectin in stable survivors with liver transplantation and healthy volunteers were assessed by enzyme-linked immunoabsordent assay (ELISA). Recently performed 6 cases of liver transplantation were also dynamically observed in this study. RESULTS: Percentage of CD4+ T cells， CD8+ T cells and CD3+ T cells, as well as ratio of CD4 to CD8 were no difference between two groups (P>0.05). However, a significant higher percentage of CD3+CD25+ T cells was found in stable liver transplantation group as compared to healthy group (P<0.05). Significantly increased concentrations of IL-6, ICAM-1 and P-selectin were found in stable liver transplantation group as compared to healthy group (P<0.05). A high TNF-α level was detected in stable liver transplantation group while no significant difference was found as compared to healthy volunteers group (P>0.05). There was not found no regular change of serum cytokines (IL-6, TNF-α) and adhesion molecules (ICAM-1, P-selectin) in 6 liver transplanted patients during post-operation from day 1 to day 30, indicating that was associated with the different status of patients before or after transplantation. CONCLUSIONS: Our data suggesting that increased levels of ICAM-1 and P-selectin, appears to participate in the processing of immunoregulation to transplanted livers, whereas elevated concentrations of IL-6 appear to be involved in the repair of the injury induced by TNF-α in allo-transplanted livers.     

Proportion of CD14+CD16+ monocytes in peripheral blood from patients with type 2 diabetic mellitus and effect of LPS and IL-15 on expression of STAT5 in monocytes

AIM: To characterize the proportion of CD14⁺CD16⁺ monocytes in peripheral blood from type 2 diabetes (T2DM) patients and to observe the response of CD14⁺CD16⁺ monocytes to lipopolysaccharide (LPS) and interleukin-15 (IL-15) for further exploring the potential mechanism of inflammatory immune response in the pathogenesis of T2DM. METHODS: Twenty-eight patients with T2DM and 20 healthy volunteers were enrolled in the study. The peripheral blood was collected for determining the percentage of CD14⁺CD16⁺ monocytes by flow cytometry. The peripheral blood mononuclear cells (PBMC) were isolated and subject to stimulation with LPS and IL-15 for 4 h. The protein expression of STAT5 was detected by Western blotting and the phosphorylated (p)-STAT5 was determined by Western blotting and immunofluorescence. Serum levels of 25-hydroxyvitamin D₃ and IL-6, and the concentrations of IL-6 and monocyte chemoattractant protein-1(MCP-1) in the culture supernatants were assessed by ELISA. Serum level of C-reactive protein (CRP) was measured by immunoturbidimetry. RESULTS: There were positive correlations between the quantity of CD14⁺CD16⁺ monocytes and serum levels of CRP and IL-6 (r=0.394, P<0.05 and r=0.741, P<0.01), while serum 25 (OH) D₃ was negatively correlated with the quantity of CD14⁺CD16⁺ monocytes (r=-0.409, P<0.01), serum CRP(r=-0.479,P<0.01) and serum IL-6 (r=-0.774,P <0.01). After stimulated with LPS and IL-15, PBMC showed significant up-regulation of p-STAT5 protein expression, and significant increases in the supernatant levels of IL-6 and MCP-1 were observed (P<0.05). The expression of p-STAT5 existed in the nucleus.CONCLUSION: These findings suggest that the functional disturbance in monocytes occurs in T2DM, which may be related to insufficiency of vitamin D₃. The aberrant activation of STAT5 signaling pathway underlies the functional abnormalities of the monocytes in T2DM.     

Significance of NF-кB in immunopathogenesis of Graves disease

AIM:To investigate the activity of NF-кB in peripheral blood mononuclear cells (PBMCs) from patients with Graves disease (GD) and the significance in immunopathogenesis of GD.METHODS:Peripheral blood was collected from 22 untreated GD, 20 treated GD with tapazole more than 1 year, and 25 healthy volunteers.PBMCs were isolated from the blood by histopaque-1077 density-gradient centrifugation.The activity of NF-кB in PBMCs was analyzed using gel electrophoretic mobility shift assay (EMSA).The contents of IL-1β, IL-6 and TNF-α were tested by radioimmunoassay.RESULTS:The activity of NF-кB in PBMCs of untreated GD group was increased remarkably, compared with that in the treated group and control (P<0.05).The contents of IL-1β, IL-6 and TNF-α in untreated group were significantly higher than those in treated GD and control group (P<0.05).A positive correlation between NF-кB activity and IL-6 level in untreated GD group and treated GD group was observed.CONCLUSION:The activity of NF-кB in PBMCs with GD patients is increased significantly, which might play an important role in the immunopathogenesis of GD.     

Alteration of serum IL-6, IL-8, TNF-α and sICAM-1 in patients with pre-menstruation recurrent aphthous ulceration

AIM:To investigate the serum levels of interleeukin-6(IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha(TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1)in female patients with pre-menstruation recurrent aphthous ulceration(RAU).METHODS:Serum levels of IL-6, IL-8, TNF-α and sICAM-1 in 21 pre-menstruation RAU patients were examined using ELISA technique, and compared to 10 healthy individuals and 22 the female RAU patients unrelated to menstrual cycle.RESULTS:The serum levels of IL-6, IL-8, TNF-α in patients with pre-menstruation RAU were not only significantly higher than that in the normal control group(P＜0.01), but also higher than that in the RAU patients without pre-menstruous recurrence (P＜0.01).The level of serum TNF-α in the RAU patients without pre-menstruous recurrence was slightly higher than that in the normal control group(P＜0.05).The level of sICAM-1 had not changed.CONCLUSION:The serum levels of IL-6, IL-8 and TNF-α increase in pre-menstruation RAU patients, which might play a role in local lesion of RAU.     

Relationship between estrogen and C-type natriuretic peptide-induced pubertal linear growth

AIM：To explore the association between estrogen and C-type natriuretic peptide (CNP)-induced endochondral ossification in pubertal girls, and to investigate the relationship between CNP signaling pathway and the gonadotropin-releasing hormone analogue (GnRHa)-associated linear growth reduction in girls with idiopathic central precocious puberty (ICPP). METHODS：Serum levels of estradiol (E2), N-terminal propeptide of CNP (NT-proCNP), insulin-like growth factor 1 (IGF-1) and N-terminal mid-fragment of osteocalcin (N-MID OC) were measured in 56 healthy girls at different pubertal stages, and in 13 girls with ICPP at the beginning, the end of the 6th and 12th months of GnRHa treatment. Height velocity (HV) was also calculated. RESULTS：(1) Serum NT-proCNP, IGF-1, E2 and N-MID OC levels in the 56 healthy girls increased at early puberty compared with prepubertal levels (P<0.05 or P<0.01). Serum NT-proCNP level remained high at middle puberty and peaked at late puberty (P<0.05). Serum IGF-1 and E2 levels continued to increase at middle puberty (P<0.01) and peaked at late puberty (P<0.01). Serum N-MID OC level peaked at middle puberty (P<0.05) and decreased at late puberty (P<0.05). (2) The HV and serum levels of NT-proCNP and N-MID OC in the 13 ICPP girls decreased at the end of the 6th month of GnRHa treatment compared with those at the beginning of GnRHa treatment (P<0.05), and remained low at the end of the 12th month of GnRHa treatment. Serum IGF-1 level at the end of the 6th and 12th months of GnRHa treatment remained as the same as that at the beginning. Serum E2 returned to the prepubertal level after GnRHa treatment (P<0.05). CONCLUSION：Serum NT-proCNP level increases across female pubertal process and peaks at late puberty, in parallel with serum E2 and IGF-1 levels. Elevated estrogen in female puberty may be associated with CNP-mediated adolescent growth spurt. Serum NT-proCNP level in ICPP girls decreases during GnRHa treatment, in parallel with linear growth velocity and bone formation. Linear growth reduction in girls with ICPP treated with GnRHa is partly due to decreased CNP-mediated long bone growth after estrogen inhibition.     

Studies on the role of ET-1 and CGRP in the pathophysiology of ankylosing spondylitis and rheumatoid arthritis

AIM: To clarify the role of endothelin-1 (ET-1) and calcitonin gene related peptide(CGRP) in the pathophysiology of ankylosing spondylitis and rheumatoid arthritis. METHOD: Plasma/synovial fluid ET-1 and CGRP were measured by radioimmunoassay in patients with ankylosing spondylitis (AS) or rheumatoid arthritis (RA) and healthy control. RESULTS: ET-1 level in plasma of patients with AS and RA were significantly higher than that of healthy controls (P<0.01). No difference was found in plasma CGRP level between AS or RA and healthy control (P>0.05). CGRP level in synovial fluid was significantly higher than that in plasma (P<0.01), but ET-1 level was significantly lower than in plasma (P<0.01). CONCLUSION: These results suggest that ET-1 and CGRP play a pathogenic role in AS and RA.     

Relationship between Th1/Th2 balance and CD28/CTLA-4 expression on peripheral blood T cells in patients with systemic lupus erythematosus

AIM: To investigate the pattern of Th1/Th2 balance in systemic lupus erythematosus(SLE) patients and the relationship between CD28/CTLA-4(cytotoxic T-lymphocyte antigen-4) molecule expression and Th1/Th2 balance.METHODS: Eighteen SLE patients met the ARA 1997 updated SLE criteria were selected in the study. According to Bombardier's SLEDAI criteria, all patients were classified into two groups: active group(12 cases) and static group(6 cases). Fourteen normal individuals, matched for age and sex of the patients, served as controls. The peripheral blood mononuclear cells(PBMCs) were isolated by density gradient centrifugation and cultured in RPMI-1640 culture medium. After treated with PMA(5 μg/L) and ionomycin(500 μg/L) for 72 h, the PBMCs were collected, the contents of IFN-γ and IL-10 in the supernatant of cultured PBMCs were detected using enzyme linked immunosorbent assay(ELISA). The expression of CD28 and CTLA-4 molecules on T cells were detected by flow cytometric technique with double staining by FITC or PE labeled monoclonal antibodies. RESULTS: The level of IL-10 was higher in the PBMCs of active and static SLE patients(351.29 ng/L±153.31 ng/L and 319.37 ng/L±153.39 ng/L) than that in controls(254.48 ng/L±120.69 ng/L), but the difference did not reach statistical significance(P>0.05). The level of IFN-γ was significantly lower in the PBMCs of active SLE patients(25.76 ng/L±16.09 ng/L) than that in controls(50.71 ng/L±27.92 ng/L, P＜0.05). The ratio of IL-10/IFN-γ was significantly higher in active SLE patients(18.74±13.77) than that in controls(6.66±4.95, P<0.05). Either before or after culture, the expression of CD28 molecule on CD3⁺and CD8⁺ T cells from all SLE patients was not remarkably different from that in the cells of controls. Before culture, the expression of CTLA-4 molecule on CD3⁺T cells of active SLE patients(0.79%+0.37%) was significantly lower than that in the cells of controls(1.31%+0.61%, P<0.05). After culture, the expression of CTLA-4 molecule on CD3⁺ T cells of SLE patients was still lower than that in the cells of normal controls without statistical significance(P＞0.05).The expression level of CD28 molecule on CD3⁺ or CD8⁺ T cells in active SLE patients and controls was not correlated with the levels of IFN-γ and IL-10 in the supernatants(P＞0.05). The level of CTLA-4 molecule expression on CD3⁺ T cells of active SLE patients was positively correlated with IFN-γ level(r=0.681, P<0.05), while was negatively correlated with IL-10 levels(r=-0.624,P＜0.05) and the ratio of IL-10/IFN-γ(r=-0.738, P<0.01). The level of CTLA-4 molecule expression on CD3⁺ or CD8⁺ T cells of controls showed no correlation with IFN-γ levels, while showed negative correlations with IL-10 level(r=-0.587, P<0.05; r=-0.563, P<0.05, respectively).CONCLUSION: There is a bias in the differentiation of Th0 cells towards Th2 in SLE patients. CTLA-4 probably plays an important role in this mechanism through suppressing the signal transmitted by CD28.     

Peripheral blood mononuclear cells activated by graded zirconia-hydroxyapatite composite in vitro

AIM: To evaluate the immunogenicity of a novel orthopedics materials (graded zirconia-hydroxyapatite composite) in vitro by using peripheral blood mononuclear cells (PBMCs) from healthy young people, and simple zirconia-hydroxyapatite composited material was used as control materials. METHODS: Proliferation of PBMCs cultured in different liquid after 5 days was measured by MTT methods. ELISA was used to detect TNF-α and IL-6 concentration in the supernatant of PBMCs cultured in the extracts after 24 hours. Flow cytometery was used to measure CD69 and CD25 in activated PBMCs cultured in the extracts of the two kinds of materials after 24 hours. RESULTS: The proliferation rate of the simple composite group was significantly lower than that in negative group (P<0.05), but there was no difference between the graded composited group and negative group. After 24 hours culture with LPS, the concentrations of TNF-α and IL-6 in the simple composited group were significantly higher than that in graded composited group, respectively (P<0.05). Cultured with PHA for 24 hours, the ratio of CD69 and CD25 positive PBMCs in the simple composited group was all significantly higher than that in graded composited material group (P<0.01). CONCLUSION: The number of PBMCs activated by the graded composited material is less than the simple composited material and the immunogenicity of the graded composited material is lower than the simple composited material.     

TLR7 agonist enhances anti-tumor activity of PBMCs from patient of renal cell carcinoma

AIM:To investigate the effect of Toll-like receptor 7(TLR7) agonist on the anti-tumor activity of peripheral blood mononuclear cells (PBMCs) in the patient with renal cell carcinoma.METHODS:Primary renal cancer cells from the postoperative specimens of the patient were co-cultured with peripheral blood mononuclear cells from the same patient stimulated by TLR7 agonist. The cytokine levels in culture medium were measured by ELISA, and the cell cycle distribution of the renal cell carcinoma cells was analyzed by flow cytometry. The anti-tumor activity of PBMCs was evaluated by[⁵¹Cr] release trial. The protein levels of Skp2 and its downstream pathway molecules were determined by Western blot. RESULTS:TLR7 agonist increased the expression of interferon-γ(IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) in the co-cultured medium, which significantly inhibited the proliferation index of the renal cell carcinoma cells. The cytotoxicity of PBMCs to renal cell carcinoma cells was markedly increased (P<0.05). The protein level of Skp2 in renal cell carcinoma cells was decreased significantly after stimulation, which was consistent with the change of cell proliferation index of renal cell carcinoma cells(P<0.05). The protein level of p27 was increased significantly (P<0.05), which was opposite to the change of Skp2. However, no significant difference in the expression of p21 and p53 was observed. CONCLUSION:TLR7 agonist effectively enhances the anti-tumor activity of PBMCs and results in the growth of renal cell carcinoma cells inhibiting. The mechanism may be relate to the cell cycle arrest by inhibiting the Skp2/p27 pathway.     

Effect of glucocorticoid-treated dendritic cells on asthmatic Th2 response

AIM: To investigate the effect of dexamethasone-treated dendritic cells (DCs) on Th2 cytokine production from autologous T cells in asthmatic patients and explore the mechanisms by studying the effect of dexamethasone on differentiation, maturation and function of DCs from patients with asthma. METHODS: Human peripheral blood monocyte-derived DCs generated from asthmatic patients and healthy subjects were cultured in the absence or presence of dexamethasone. The phenotypic characterization of DCs was analyzed by flow cytometry. The mature DCs were harvested, washed, and then cocultured in vitro with autologous T cells purified by a nylon cotton column. The DC-T coculture supernatants were collected after 72 h incubation and analyzed for levels of IL-5 and IFN-γ by ELISA. RESULTS: The concentrations of IL-5 in the culture supernatants of DC-T coculture were significantly up-regulated in patients with asthma compared with that in healthy controls ［(145.13±89.76) ng/L vs (50.28±22.37) ng/L, P<0.01］. The level of IFN-γ in the DC-T coculture supernatants tended to be decreased in asthmatic patients than that in healthy controls, although this difference did not achieve statistical significance ［(197.58±76.32) ng/L vs (220.46±65.34) ng/L, P>0.05)］. There were significantly decreased levels of IL-5 by autologous T cells primed by dexamethasone-treated mature DCs from asthmatic patients ［(45.39±19.61) ng/L vs (145.13±89.76) ng/L, P<0.01］, alterations not observed from healthy controls (P>0.05). IFN-γ production was decreased by autologous T cells primed by dexamethasone-treated mature DCs from both asthmatic patients and healthy controls ［asthma group: (40.21±22.89) ng/L vs (197.58±76.32) ng/L, P<0.01; healthy controls: (56.78±20.37) ng/L vs (220.46±65.34) ng/L, P<0.01］. Dexamethasone-treated DCs exhibited decreased expression of CD83 (P<0.01) and increased expression of CD14 (P<0.01) in both asthmatic patients and healthy controls. CONCLUSION: DCs of asthmatic patients induce a Th2-skewed cytokine production from autologous T cells. Dexamethasone-treated DCs inhibit the Th2 reactions, and this effect is probably mediated through the pathway that dexamethasone inhibits DCs maturation and skews the macrophage/DC balance towards the macrophage side and thus directs the development more towards the macrophage lineage.     

Change of intestinal flora and its relationship with IL-23/IL-17 axis in patients with ulcerative colitis

AIM:To investigate the change of intestinal flora distribution and its relationship with interleukin-23 (IL-23)/IL-17 axis in ulcerative colitis (UC) patients. METHODS:The fresh fecal samples from 20 patients with active UC and 20 healthy controls were collected. The distribution of the flora was analyzed by direct smear and traditional bacterial culture. The changes of bacteria were detected by real-time PCR. The hemoglobin, albumin, erythrocyte sedimentation, and C-reactive protein levels were tested routinely. Both normal and damaged mucosal tissues of UC patients were examined and obtained by colonoscopy, and further assessed by Mayo scoring, Baron grading and HE staining. The expression of IL-17 and IL-23 was observed by immunohistochemistry and Western blot. RESULTS:(1) The degree of flora imbalance in active UC patients was higher than that in the healthy controls (P<0.05). (2) The results of aerobic culture showed that the number of Escherichia coli in the UC patients was significantly lower than that in the normal controls (P<0.01), while Enterococcus was increased obviously (P<0.01). The results of anaerobic culture revealed that the numbers of Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients were significantly decreased (P<0.01). (3) Quantitative analysis of target bacteria showed that the relative quantification of Escherichia coli, Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients was significantly lower than that in the normal subjects, and the number of Enterococcus was significantly increased (P<0.01). (4) Compared with control group, no significant change of hemoglobin in the UC patients was ovserved, albumin was significantly decreased (P<0.05), but erythrocyte sedimentation and C-reactive protein levels were elevated obviously (P<0.01). (5) The Mayo score, Baron grade, and histopathological score were all increased (P<0.01). (6) High IL-17 and IL-23 expression levels were detected in the UC patients (P<0.01). (7) Correlation analysis showed that the average absorbance values of IL-17 and IL-23 expression were positively correlated with Baron grade (r=0.717, P=0.02; r=0.849, P=0.016) and pathological score (r=0.660, P=0.03; r=0.675, P=0.032). Meanwhile, the average absorbance value of IL-23 expression was negatively correlated with the number of Escherichia coli (r=-0.699, P=0.025), and positively correlated with Enterococcus (r=0.872, P=0.010). Furthermore, the average absorbance value of IL-17 expression was positively correlated with Enterococcus (r=0.764, P=0.046), and both of them were not correlated with other bacteria. CONCLUSION:Obvious flora imbalance exists in active UC patients, changed intestinal microflora is closely related with the degree of inflammation. IL-23/IL-17 axis, as a key factor in the development of UC, may be related to the changes of intestinal microflora. The interaction between intestinal microflora and IL-23/IL-17 axis plays an important role in the pathogenesis of UC.     

Preliminary study of IL-17 signal transduction pathway in peripheral blood mononuclear cells from lupus nephritis patients

AIM: To investigate the role of IL-17 and its signal conduction component-JNK activity in the pathogenesis of LN. METHODS: Peripheral blood mononuclear cells (PBMC) were separated and cultured from 15 cases of active lupus nephritis (LN) patients. IL-6 level was detected by ELISA, IL-6 mRNA was checked with RT-PCR, and JNK activity was measured by Western blot. RESULTS: At same IL-17 end concentrations, there was a much higher level of IL-6 in LN group than in control group (all P<0.05). IL-17 induced a significant elevation of IL-6 mRNA expression and JNK activity in PBMC from LN patients in a time- and dose-dependent manner, which could be blocked markedly by IL-17 monoclonal antibody, mIgG₂₈, and dexamethasone. Much higher IL-6 mRNA expression was observed in LN group than in control group under medium culture or IL-17-conditioned culture (all P<0.01). Under medium culture or IL-17-conditioned culture, there was much higher JNK activity of PBMC in active LN group than that in control group. There was a positive linear correlation between PBMC JNK activity and their SLEDAI or IL-6 mRNA expression level in LN patients (r1=0.638, P<0.01; r2=0.644, P<0.01). CONCLUSION: These results suggest that IL-17 may take part in the initiation and progression of LN through induction of IL-6 overexpression by PBMC,and JNK hyperactivity may be necessary in the IL-17 signal transduction.     

IL-33/ST2 axis in systemic lupus erythematosus in relation to chronic kidney injury and disease activity

AIM: To elucidate the association between chronic kidney injury and interleukin-33 (IL-33; an alarmin)/suppression of tumorigencity 2 (ST2) in patients with systemic lupus erythematosus (SLE).METHODS: Serum levels of IL-33 and soluble ST2 (sST2) were assessed by ELISA in 50 SLE patients and 30 healthy controls (HC).RESULTS: The levels of IL-33 and sST2, and IL-33/sST2 ratio were significantly higher in SLE patients than those in the HC. The IL-33 and sST2 levels were positively associated with SLE disease activity index (SLEDAI), erythrocyte sedimentation rate (ESR), proteinuria and triglyceride, but negatively associated with complement C3. IL-33/sST2 ratio was positively associated with SLEDAI and estimated glomerular filtration rate (eGFR). Independent explanatory variables associated with high IL-33/sST2 included chronic kidney disease (CKD) staging and albumin (R²=0.442), especially CKD staging.CONCLUSION: Elevated serum sST2 and IL-33 levels in SLE patients are correlated with disease activity and risk factors of kidney injury. IL-33/sST2 ratio may serve as a potential biomarker for chronic kidney injury in SLE patients.     

Correlation of intestinal endotoxemia,histaminemia and cellular immune function in patients with hepatitis B

AIM: To investigate the correlation of intestinal endotoxemia (IETM), histaminemia and cellular immune function in the patients with hepatitis B. METHODS: Peripheral blood was collected from patients with chronic hepatitis B (n=80) and healthy individuals (n=18). According to plasma endotoxin concentration, total patients were divided into two groups: ET positive and ET negative. Serum IL-10, IL-12, IFN-γ, IL-2, IL-4 concentrations were detected. In addition, the serum histamine (HA), tryptase (TS) and AP50 levels were studied. RESULTS: Compared to control group, the concentrations of IL-4 and IL-10 were increased, but IL-12 and IFN-γ were decreased obviously in total patients (P<0.05). The levels of IL-4 and IL-10 in ET positive group were higher than that in ET negative group (P<0.05). The level of IL-12 and IFN-γ had no statistical difference between two groups. AP50, HA and TS levels were increased significantly in total patients compared with control group, and the levels of ET positive were higher than that in ET negative group (P<0.05). Furthermore, in total patients, ET was correlated with IL-4, IL-10, AP50 and HA, respectively. HA was negatively correlated with IL-12 and IFN-γ, and correlated with AP50 and TS. In ET positive group, ET was correlated with IL-4, IL-10, AP50 and HA, respectively, which did not exist in ET negative group. CONCLUSION: AP50 may be a sign of IETM. IETM may be disbenefit to hepatitis B through activating complement, which leads to histaminemia and poor cellular immune function.     

Ulinastatin protects rat pulmonary tissues from paraquat-induced acute injury

AIM: To investigate the protective effects of ulinastatin on the rats with paraquat-induced acute lung injury and its mechanisms. METHODS: The Wistar rats (n=108) were randomly divided into control group, paraquat group and ulinastatin group. The rats in paraquat group and ulinastatin group were given paraquat by gavage, while the rats in control group were given sterile saline by gavage. The rats in ulinastatin group were also given ulinastatin treatment. The serum levels of MDA, SOD, IL-6, IL-10 and TNF-α were measured after 1 d, 3 d, 7 d, 14 d, 21 d and 28 d. The expression levels of p38 MAPK, MMP-2 and TIMP-1 in the lung were also measured. RESULTS: The levels of SOD in 1 d, 3 d and 7 d in paraquat group and ulinastatin group were significantly lower than those in control group (P<0.01). The level of SOD in ulinastatin group was significantly higher than that in paraquat group (P<0.05). The levels of MDA, IL-6, IL-10 and TNF-α in 1 d, 3 d and 7 d in paraquat group and ulinastatin group increased compared with control group (P<0.01), and those in ulinastatin group were significantly lower than those in paraquat group (P<0.05). The levels of p38 MAPK and TIMP-1 in 1 d, 3 d, 7 d, 14 d, 21 d and 28 d in paraquat group and ulinastatin group were higher than those in control group (P<0.01), and those in ulinastatin group was significantly lower than those in paraquat group (P<0.05). The level of MMP-2 in 1 d, 3 d, 7 d, 14 d and 21 d in paraquat group and ulinastatin group increased compared with control group (P<0.01), and that in ulinastatin group was significantly lower than that in paraquat group (P<0.05).CONCLUSION: Ulinastatin protects the lung tissues of rats from paraquat-induced acute lung injury by inhibiting p38 MAPK signaling pathway and ameliorating inflammatory and oxidative responses.