A study on the expression of FGF-21 and NF-κB pathway in the tissues of atherosclerotic mice.

OBJECTIVE: To study the relationship between the expressions of fibroblast growth factor (FGF)-21 and NF-κB signal transduction pathway in the tissues of atherosclerotic mice. MATERIALS AND METHODS: A total of 40 apoE-/- male mice at 8 weeks were selected and randomly divided into 4 groups. 10 mice in group A were normally fed with diet. 10 mice in group B were fed with high-fat diet. 10 mice in group C were fed with high-fat diet + pravastatin. 10 mice in group D were fed with high-fat diet + subcutaneous injection of exogenous recombinant FGF-21 protein. Another 10 C57BL/6J mice at 8 weeks were normally fed with diet (group E). They were killed after 12 weeks to collect retinal venous blood. ELISA method was applied to detect the levels of serum FGF-21, NF-κB, monocyte chemo attractant protein (MCP-1), matrix metalloproteinase (MMP)-9 and TNF-α. Immunohistochemical staining and RT-PCR method were applied to detect the expression of FGF-21 in aortic arch and liver tissues. RT-PCR method and Western blot method were applied to detect the expression of NF-κB, MCP-1, MMP-9 and TNF-α in aortic arch and liver tissues. RESULTS: The levels of serum FGF-21, NF-κB, MCP-1, MMP-9 and TNF-α in group B were higher than those of group A and group E, and those of group C and group D were lower than those of group B (except FGF-21 in group D). The differences had statistical significance (p<0.05). The positive staining rates of FGF-21 in endothelial cells of aortic arch and liver tissues in group B were higher than those group A and group E, and those of group C and group D were lower than those of group B. The differences had statistical significance (p<0.05). The expression levels of FGF-21mRNA, NF-κB, MCP-1, MMP-9, TNF-αmRNA and protein in endothelial cells of aortic arch and liver tissues in group B were higher than those group A and group E, and those of group C and group D were lower than those of group B. The differences had statistical significance (p<0.05). CONCLUSIONS: FGF-21 may participate in the occurrence of atherosclerosis (AS), which is related to the activation of the NF-κB pathway. Lipid-lowering therapy can inhibit the activation of FGF-21 and NF-κB. Exogenous FGF-21 can also lower the activation of NF-κB and interpose in atherosclerosis process.

Increased level of MSU crystal-bound protein apolipoprotein A-I in acute gouty arthritis.

BACKGROUND: Gout is a common form of inflammatory arthritis that is triggered by the crystallization of monosodium urate (MSU). We investigated the potential proteins that relate to the pathogenesis or the spontaneous resolution of acute gouty arthritis. METHOD: We screened for differentially expressed proteins in the plasma of patients with acute gouty arthritis using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) identification. We confirmed these findings in a population study of 209 subjects, and further determined the protein profile of the synovial fluid (SF) from 24 gouty patients during acute attack by liquid chromatography coupled with tandem MS (LC/MS/MS). RESULTS: The highly expressed apolipoprotein A-I (apoA-I) was identified in the plasma of acute gouty patients compared with healthy controls. Moreover, we detected high levels of SF apoA-I in 83.3% of acute gouty patients during attack. From the population study, apoA-I was increasingly associated with normouricaemia, hyperuricaemia, and acute gouty arthritis (ptrend < 0.001), and plasma uric acid (UA) and apoA-I were positively correlated (p = 0.0156). We used a human liver cell model and found that UA enhanced the hepatic apoA-I mRNA expression level (ptrend < 0.01) and apoA-I secretion level (ptrend = 0.002) in a dose-dependent manner. An elevated MSU concentration caused the endogenous apoA-I to deplete gradually. CONCLUSIONS: Based on the role of apoA-I in anti-inflammation, our observational data in acute gout support the hypothesis that apoA-I expression can be induced under the condition of a high concentration of UA and its elevated level may be implicated in the spontaneous resolution of acute gouty arthritis.

Defective Mycobacterium tuberculosis antigen presentation by monocytes from tuberculosis patients.

OBJECTIVE: To determine whether there were any defects in the presentation of the Mycobacterium tuberculosis antigen by monocytes from patients with tuberculosis (TB) and the role of vitamin D in the defence against M. tuberculosis. DESIGN: A prospective study aimed at analysing the presentation of the M. tuberculosis antigen by monocytes and the response to vitamin D treatment in three groups of participants: 1) those with active TB, 2) those with healed TB and 3) those with frequent TB contact. RESULTS: The antigen presentation ability of monocytes of persons in the frequent contact group was significantly higher than that of the active TB and healed TB groups. There was no difference between patients with active and healed TB. In addition, 1,25(OH)2D3 increased the presentation of mycobacterial antigens by monocytes from participants with frequent TB contact, but not those with active or healed TB. CONCLUSION: Patients with active and healed TB exhibit defective M. tuberculosis presentation in monocytes. The administration of vitamin D did not correct this defect in monocytes from participants with active or healed TB, but could increase antigen presentation by monocytes in participants with frequent TB contact.

Global DNA methylation, DNMT1, and MBD2 in patients with systemic lupus erythematosus.

To investigate the associations of DNA methylation levels and mRNA expressions of DNA cytosine-5-methyltransferase 1 (DNMT1) and methyl CpG-binding domain 2 (MBD2) with systemic lupus erythematosus (SLE), 108 patients with SLE and 97 healthy controls were enrolled in this study. DNA and total RNA were extracted from the peripheral blood mononuclear cells of the SLE patients and the controls. The global methylation levels of DNA were measured in 63 patients with SLE and 68 healthy controls by the ELISA method. DNMT1 and MBD2 mRNA were also detected in 108 SLE patients and 97 controls using the quantitative real-time polymerase chain reaction method. The global methylation level of DNA was significantly decreased in the SLE patients in comparison with that in the controls (p < 0.001, 95% CI = 0.1573-0.5052). The patients with SLE have higher expressions of DNMT1 and MBD2 mRNA than the controls (p < 0.001, 95% CI = -0.0049 - -0.0019 and p = 0.001, 95% CI = -0.0119 - -0.0029, respectively). We also found that there were no significant differences in the methylation level and the expression of DNMT1 and MBD2 mRNA between the active and the inactive SLE patients. A positive correlation was also found between DNMT1 and MBD2 mRNA expressions in the SLE patients (p < 0.001). This study demonstrated that the patients with SLE had a significantly lower level of DNA methylation than the controls. The expression of both DNMT1 and MBD2 mRNA was significantly increased in the SLE patients compared with the controls. This study also showed a positive correlation between DNMT1 and MBD2 mRNA levels in the patients with SLE.

IkappaBalpha promoter polymorphisms in patients with rheumatoid arthritis.

To investigate the role of inhibitor of kappaBalpha promoter polymorphisms in the pathogenesis of rheumatoid arthritis (RA), 140 patients with RA and 115 healthy controls were enrolled in this study. The IkappaBalpha promoter polymorphisms were determined using the polymerase chain reaction/restriction fragment length polymorphisms method. In comparison with IkappaBalpha-826 C/C, the genotype frequency of IkappaBalpha-826 C/T was significantly higher in the patients with RA than that of the controls (P = 0.009, OR = 2.0, 95% CI = 1.2-3.4). The allele frequency of IkappaBalpha-826 T was also significantly increased in patients with RA when compared with that of the controls (P = 0.027, OR = 1.6, 95% CI = 1.1-2.4). In comparison with IkappaBalpha-550 A/A, the genotype frequency of IkappaBalpha-550 A/T was significantly decreased in patients with RA when compared with that of the controls (P = 0.02, OR = 0.2, 95% CI = 0.06-0.8). The allele frequency of IkappaBalpha-550 A was significantly increased in patients with RA (P = 0.007, OR = 5.1, 95% = 1.4-18.2). This study also revealed that the IkappaBalpha-826 T -550 A -519 C haplotype was significantly increased in patients with RA in comparison to that of controls (P = 0.01, OR = 1.8, 95% CI = 1.1-2.8). The IkappaBalpha-826 T and -550 A alleles are associated with susceptibility to RA. Moreover, the IkappaBalpha-826 T -550 A -519 C haplotype is associated with susceptibility to RA in Taiwan.

Killer cell immunoglobulin-like receptor gene's repertoire in rheumatoid arthritis.

OBJECTIVE: To investigate the role of the killer cell immunoglobulin-like receptor (KIR) gene's repertoire in the pathogenesis of rheumatoid arthritis (RA) in Taiwan. METHODS: KIR genotypes were determined in 122 patients with RA and 96 healthy controls by the sequence-specific primer polymerase chain reaction (SSP-PCR) method. Human leucocyte antigen (HLA)-C genotyping was also performed simultaneously in 72 patients and 66 controls by the SSP-PCR method. RESULTS: The total carriage frequency of KIR 2DS4 regardless of corresponding HLA-Cw4 was significantly increased in RA patients compared with controls [p<0.001, odds ratio (OR) = 1.9, 95% confidence interval (CI) = 1.1-3.4, Pc<0.01]. The total carriage frequency of KIR 2DL1 regardless of corresponding HLA-C also tended to be increased in RA patients (p<0.02, OR = 2.1, 95% CI = 1.2-3.9, Pc = not significant). The frequency of KIR 2DS4 with corresponding HLA-Cw4 was increased in RA patients in comparison with controls (p = 0.02, OR = 3.2, 95% CI = 1.1-9.4). Moreover, the association of RA with KIR 2DS4 depended on the presence of the corresponding HLA-Cw4. CONCLUSIONS: KIR 2DS4 may be a risk factor for susceptibility to RA in Taiwan.

Community-based study in Taiwan aborigines concerning renal dysfunction in gout patients.

OBJECTIVE: To reveal the factors associated with renal dysfunction among gout patients in Taiwan aborigines. METHODS: Social demographic data, alcohol consumption data, anthropometric measurements, blood samples, and 24-h urine samples were collected from 128 aboriginals (101 men, 27 women) suffering from gout. RESULTS: The men displayed higher mean creatinine clearance (Ccr) values than women. Twenty-two post-menopausal women had significantly lower Ccr values compared to the five pre-menopausal women [probability (p)<0.001]. The males displayed higher 24-h urinary creatinine value than females (8.60+/-5.39 versus 5.58+/-2.14 mmol/L; p<0.05), and showed a significantly higher positive relationship between 24-h urinary creatinine and uric acid excretion [correlation coefficient (r)=0.7304; p<0.001], whereas the females did not (r=0.1144; p=0.5691). Overall, those who were older members of the Tsou tribe, or had excreted less uric acid from urine in 24 h tended to suffer renal dysfunction. CONCLUSIONS: Gout patients displayed diversity in renal function. An exogenous source of creatinine in men was more likely than in women.

Cytochrome p450 1Al gene polymorphisms in patients with psoriatic arthritis.

OBJECTIVE: To investigate the associations of cytochrome p450 1A1 (CYP1A1) gene polymorphisms with susceptibility to psoriatic arthritis in Taiwan. METHODS: CYP1A1 gene polymorphisms were determined in 52 patients with psoriatic arthritis and in 90 healthy controls by polymerase chain reaction/restriction fragment length polymorphism. RESULTS: The genotype frequency of CYP1A1 4889A/G was significantly increased in patients with psoriatic arthritis in comparison with healthy controls. The phenotype frequency of CYP1A1 4889G was also significantly increased in patients with psoriatic arthritis. The genotype frequency of CYP1A1 4887C/A was significantly higher in patients with psoriatic arthritis than in controls. The allele and phenotype frequencies of 4887A were also significantly increased in patients with psoriatic arthritis. We also found that the association of CYP1A1 4887A with psoriatic arthritis was independent of 4889G but a synergistic effect was present between CYP1A1 4887A and 4889G. The CYP1A1 4889A/G and 4887C/A polymorphisms were not associated with the manifestations and severity of psoriatic arthritis. CONCLUSION: CYP1A1 4887A and 4889G may be precipitating factors for susceptibility to psoriatic arthritis in Taiwan. An additive effect was found between CYP1A1 4887A and 4889G.

Tumor necrosis factor promoter polymorphisms in patients with rheumatoid arthritis in Taiwan.

OBJECTIVE: To investigate the association of tumor necrosis factor (TNF) promoter polymorphisms with rheumatoid arthritis (RA) in Taiwan. METHODS: TNF promoter polymorphisms at positions -238, -244, -308, -376, -857, and -863 were determined in 97 patients with RA and 97 healthy controls using the PCR-RFLP method. RESULTS: The phenotypic frequency of TNF-308A was significantly lower in patients with RA than in healthy controls. This finding can only be found in HLA-DR4 negative patients, not in DR4 positive RA patients and controls. The TNF promoter polymorphisms at positions -238, -244, -308, -376, -857, and -863 were not related to the clinical manifestations of RA patients. CONCLUSION: TNF-308A itself or a neighboring gene may be a protective factor for the development of RA in the HLA-DR4 negative population in Taiwan. TNF promoter polymorphisms were not associated with the clinical manifestations of patients with RA in Taiwan.

HLA-DQA1 genotyping in patients with rheumatoid arthritis in Taiwan.

To investigate the role of HLA-DQA1 genotypes and their interaction with HLA-DRB1 in the pathogenesis of rheumatoid arthritis (RA) in Taiwan, HLA-DQA1 was determined in 71 patients with RA and 108 healthy controls by SSP-PCR method. HLA-DRB1 and HLA-DQA1 were simultaneously detected in 55 RA patients and 101 healthy controls. PCR/SSOP method was used to determine the HLA-DRB1 genotypes, and the subtypes of HLA-DR4 were determined by cloning and sequencing. The phenotypic frequency of HLA-DQA1*0301 was significantly lower in RA than in controls, and, in contrast, the HLA-DQA1*0302 and DQA1*0303 were significantly higher in RA than in controls. The associations of DQA1*0301, *0302, and *0303 with RA were independent of DR4 and DRB1*0405. Moreover, the interactions between HLA-DR4 and HLA-DQA1*0302 or DQA1*0303 could enhance the development of RA. We also found that the prevalence of bone erosion and seropositivity of rheumatoid factor (RF) were significantly higher in HLA-DQA1*0303 positive RA patients than in healthy controls. HLA-DQA1*0302 and DQA1*0303 are the risk factors for susceptibility to RA, while HLA-DQA1*0301 is a protective factor. A synergistic effect for the susceptibility to RA can be found between HLA-DR4 and HLA-DQA1*0302 or DQA1*0303. We also found that the HLA-DQA1*0303 was related to bone erosion and seropositivity of RF in RA patients.

Familial Mediterranean fever in a Taiwanese patient.

Familial Mediterranean fever is a rare disease characterized by cyclic attacks of fever, serositis and strong family background. Here we report a 22-year-old man who suffered from recurrent fever accompanied by chest and abdominal pain for more than 10 years. The attack frequency was about once per 2-3 weeks. Although he consulted many clinics and even received appendectomy at the age of 15, no definite diagnosis was given. During the admission, many laboratory examinations failed to show any abnormality except mild leukocytosis and elevated C-reaction protein. Image studies including chest X ray and abdominal CT scan showed negative result but, interestingly, Gallium-67 scan showed a hot spot in right lower chest and right lower abdomen. After prophylaxis with colchicine 1.0 mg per day, he has enjoyed more than 2 years without the above symptoms.

Bloodletting acupuncture of the engorged vein around Bl-40 (Wei-Chung) for acute lumbar sprain.

Bloodletting acupuncture is one of the most classic methods of acupuncture therapy, and is still popularly used to treat acute lumbar sprain in the oriental world. However, most physicians in the western world are not familiar with bloodletting acupuncture, though they may know ordinary acupuncture well. Furthermore based on the literature reviewed, there have been few studies which have investigated the effect of bloodletting acupuncture upon acute lumbar sprain. In this study, we tried to determine if bloodletting acupuncture is effective for acute lumbar sprain. In total, twelve patients were enrolled for analysis. Five patients were treated with ordinary acupuncture upon the contralateral SI-3 (Hou-Hsi) point alone. Seven patients were first treated with bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 (Wei-Chung), and then followed by ordinary acupuncture upon the contralateral SI-3. It was demonstrated that bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 followed by ordinary acupuncture upon the contralateral SI-3 had more pain relief than ordinary acupuncture upon the contralateral SI-3 alone (83 +/- 23% vs. 44 +/- 28%) (P < 0.01). And bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 decreased pain by 56 +/- 23%, similar to that of ordinary acupuncture upon the contralateral SI-3 alone (44 +/- 28%). These findings suggest that bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 (Wei-Chung) has a substantial contribution for treatment of acute lumbar sprain.

Systemic lupus erythematosus complicated by recurrent spontaneous pneumothorax--a case report.

Despite frequent pleuro-pulmonary involvement, spontaneous pneumothorax is rare in patients with systemic lupus erythematosus (SLE). Here, we report a 17-year-old female patient with SLE, complicated by multiple organs involvement. She initially presented with interstitial pneumonitis and pulmonary hemorrhage, followed by spontaneous pneumothorax and CNS involvement. The patient was treated with immunosuppressive agents, including steroid and cyclophosphamide pulse therapies. Spontaneous pneumothorax happened to her once again at a different location during treatment. After aggressive therapy, the disease activity of SLE gradually diminished, and pneumothorax had not recurred during the further follow-up. Therefore, the pneumothorax may occur in SLE patients with high disease activity.

HLA-DMA and HLA-DMB genotyping in patients with systemic lupus erythematosus.

UNLABELLED: OBJECTIVE. To investigate the relation of HLA-DMA and DMB alleles to the development of systemic lupus erythematosus (SLE). METHODS: We studied HLA-DMA and DMB alleles in 108 patients with SLE and 138 healthy controls using the amplification created restriction site method. RESULTS: The phenotypic frequency of HLA-DMA*0104 was significantly higher in patients with SLE than in healthy controls. The HLA-DMA and DMB alleles were not related to clinical manifestations and various laboratory tests in patients with SLE. CONCLUSION: HLA-DMA*0104 may be associated with the susceptibility of SLE.

Identification of a new single nucleotide substitution on the hypoxanthine-guanine phosphoribosyltransferase gene (HPRT(Tsou) from a Taiwanese aboriginal family with severe gout.

OBJECTIVE: A new single nucleotide change of the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene on coding region has been identified from a Taiwanese aboriginal family with gout. The mutation was used to screen 27 members of the family, 22 Tsou, 70 Atayal, and 76 Bunun children, the elders for whom had been found to have a high prevalence of gout. METHODS: An entire peptide of HPRT coding region was directly sequenced from the cDNA of a patient with severe gout, and by using polymerase chain reaction and restriction fragment length polymorphism to screen the other participants. RESULTS: A new nucleotide change located at nucleotide 152 (G to A transition) was found that predicted an arginine to glutamine substitution at amino acid 51. This variant was named HPRT(Tsou), and was found in 3 women and 3 men among the patient's 7 siblings, 2 boys and 2 girls among the 8 children of the siblings, and one female Tsou (4.5%, 1/22) and one female Atayal (1.4%, 1/70). The serum uric acid concentration among male HPRT(Tsou) carriers in the patient's family was significantly higher than among those who had at least one HPRT gene that did not have HPRT(Tsou). CONCLUSION: We found that the HPRT(Tsou) gene variant is partially responsible for the hyperuricemia in an aboriginal population in Taiwan known for a high incidence of gout.

HLA-DMA and HLA-DMB genotyping in patients with rheumatic diseases.

To investigate the correlation of HLA-DMA and DMB alleles to some rheumatic diseases, HLA-DMA and DMB genes were detected in 11 patients with juvenile rheumatoid arthritis (JRA), 22 patients with psoriatic arthritis, 26 patients with Behcet's disease, 62 patients with ankylosing spondylitis (AS), and 138 unrelated healthy controls. There was no significant difference in phenotypic frequencies of HLA-DMA and DMB alleles between controls and patients with these rheumatic diseases. HLA-DMA and DMB genes are not related to the susceptibility of JRA, psoriatic arthritis, Behcet's disease, and AS.

Genetic analysis of interleukin-10 promoter region in patients with systemic lupus erythematosus in Taiwan.

Overproduction of interleukin-10 (IL-10) may play an important role in the development of systemic lupus erythematosus (SLE) or lupus nephritis. There is also a polymorphic dinucleotide repeat in the human IL-10 promoter region (IL-10PR). Our aim was to study whether or not the IL-10PR alleles contributed to the susceptibility to SLE or lupus nephritis. One hundred SLE patients and 103 healthy controls were studied for IL-10PR by PCR and electrophoretic analysis. The distribution of IL-10PR alleles, genotypes and the sum of both alleles (SBA) from different groups or subgroups were analyzed. SLE patients showed no difference in the distribution of IL-10PR alleles, genotypes and SBA, as compared to healthy controls. Lupus nephritis patients (N = 49) also showed no difference in IL-10PR alleles, genotypes and SBA, as compared to SLE patients without nephritis (N = 51). Of 49 lupus nephritis patients, ten developed end-stage renal disease (ESRD) and four of them were found to suffer from rapid progressive renal failure (RPRF). Patients with RPRF presented much smaller SBA than other ESRD patients (p = 0.005). Lupus nephritis patients carrying small SBA (< 18) suffered from a higher prevalence of RPRF than lupus nephritis patients without small SBA (50% V.S. 0%, p < 0.001, relative risk 82). Our data provide the first evidence of a strong association between IL-10PR and severe progression of lupus nephritis in human patients. In the future, a prospective genetic analysis of IL-10PR for patients with lupus nephritis is recommended. It might be helpful for physicians to identify the lupus nephritis subgroup with a high risk of developing RPRF early, because this might lead to a better therapy and prognosis for these patients.

T cell receptor gene V alpha and V beta usage in patients with rheumatoid arthritis in Taiwan.

To investigate the correlation of T cell receptor (TCR) genes to rheumatoid arthritis (RA) in Taiwan, synovial fluid and peripheral blood were examined simultaneously in 14 patients with RA, and peripheral blood only was examined in 15 healthy controls. RNA was extracted from T cell in synovial fluid and peripheral blood, and cDNA was synthesized using the reverse transcription method. Then, TCR-V alpha and V beta gene families were determined using the polymerase chain reaction and slot blot methods. The numbers of TCR-V alpha and V beta families, especially V beta, were decreased in rheumatoid synovial fluid compared with numbers found in the peripheral blood of the same RA patients. TCR-V beta 7 and V beta 8 could be found in the synovial fluid of all patients with positive HLA-DR4. However, there was no significant difference in the numbers of TCR-V alpha and V beta families in peripheral blood of RA patients and the peripheral blood of healthy controls. In conclusion, restricted heterogeneity of TCR-V gene, especially V beta, can be found in synovial fluid of patients with RA. TCR-V beta 7 and V beta 8 may be related to the pathogenesis of HLA-DR4 positive RA patients in Taiwan.