Objective　To observe the effect of mycophenolate mofetil (MMF),a new type of immunosuppressant,on the immune function in patiens with systemic lupus erythematosus (SLE).Methods　The changes of serum lL-6,IL-8,TNF-α,sIL-2,ANA,A-dsDNA and subsets of lymphocytes were observed in patients with SLE before and after treatment with MMF and CTX by ELISA,indirect immunofluorescent assay and flow cytometry,and the effect of MMF and CTX on the functions of hematopoiesis,liver and kidney were also observed.Results　Three months after treatment with MMF,the serum levels of IL-6,IL-8,TNF-α,sIL-2,ANA,and A-dsDNA in patients with SLE were significantly decreased,CD3＋,CD4＋ and CD4＋CD45RA＋ cells were significantly increased,and CD8＋,CD4＋CD45RO＋,CD8＋CD45RA＋,CD8＋CD45RO＋ cells were significantly decreased.No remarkable injury was observed on the functions of hematopoiesis, liver and kidney after treatment with MMF.Conclusion　MMF can inhibit T and B lymphocytes selectvely and is a new type of immunosuppresant to treat SLE with less toxicity and side-effect.

Objective　To further investigate the balance (relative presence) of Th1 and Th2 subsets at the sites of rheumatoid inflammation,and to understand how about the expression of IL-18 in patients with rheumatoid arthritis (RA),and what the relationship exists between expression of IL-18 and rate of Th1/Th2 in RA,and between IL-18 level and activity of the disease.Method　Expression of IFN-γ,IL-4,and IL-18 mRNA in peripheral blood mononuclear cells (PBMC) from 16 patients with RA and 15 healthy subjects was detected by semi-quantitative RT-PCR.Results　① PBMC from patients with RA was found to contain greater levels of IL-18 mRNA than that from healthy subjects (P＜0.01).② IL-18 mRNA levels were correlated with IFN-γ mRNA (relative coefficients:r＝0.836,P＜0.05).③ IL-18 mRNA levels were associated with the disease activity as assessed by levels of erythrocyte sedimentation rate (r＝0.753,P＜0.05).Conclusion　IL-18 is among a matrix of inflammatory cytokines produced abundantly in patients with RA and is associated with induction of IFN-γ and activity of the disease.

Prognosis and treatment of primary Sj?gren′s syndrome with renal tubular acidosis

Objective　To better understand the clinical characters of juvenile onset spondyloarthropa-thies (JSpA).Methods　The clinical and laboratory data of 190 in-patients with JSpA were analyzed and the diagnosis,classification and differentiation of juvenile onset arthritis were discussed.Results　Among these 190 patients,163 were male,with a male to female ratio 6∶1.Of them 92.1% had the disease after the age of 8.Peak of age at onset was 12 to 15 years;157(82.6%) patients had peripheral arthritis and only 23(12.1%) patients felt low back pain at onset.During the disease course, peripheral arthritis was found in 187(98.4%) patients and the history of low back pain or buttock pain was recorded in 123(64.7%).The interval between peripheral arthritis and low back pain was from 0 to 20 years,with an average of (3.2±4.5)years.Extra-articular features including enthesitis in 67(35.3%)patients,dactylitis in 20(10.5%),iritis in 9(4.7%) were observed.HLA-B27 was positive in 87.9%(160/182) patients.Sacroiliitis on X-ray was observed in 76.0%(136/179) patients,and 106(55.8%) patients were diagnosed juvenile ankylosing spondylitis (JAS) according to 1984 New York modified criteria.The average disease course in JAS was (6.3±6.2) years,longer than that in JSpA (P＜0.01).Conclusion　The concept of JSpA is helpful to early diagnosis and treatment of juvenile onset arthritis.The JSpA are characterized by asymmetric lower limb predominant oligoarthritis,a wide spectrum of extra-articular features,presence of HLA-B27 and familial history of SpA or psoriasis.It will take an average of 6.3 years for JSpA patients to fulfill the diagnostic criteria of adult AS.

Objective　To study on effects of mVEGF and anti-VEGF antibody during cultured HUVEC proliferation in vitro.Methods　Endothelial cell proliferation was assayed using human umbilical vein endothelial cells stimulated with mVEGF and with CIA joint extracts and was used　　3　　H-TaR incorporation.Results　The anti-VEGF neutralizing antibody can inhibit the proliferation of HUVEC stimulated with mVEGF and with CIA joint extracts,whose suppression percents were 72.2% and 69.9%,respectively.Conclusion　mVEGF specifically promotes the growth of vascular endothelial cells.During early stage of CIA development,expression of VEGF in the joint increases and VEGF is expressed biologically active and can be inhibited by anti-VEGF neutralizing antibody.

Objective　To measure the levels of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in synovial fluid and plasma of patients with RA and investigate its clinical significance.Methods　By using ELISA sandwich method,the levels of uPA and uPAR in plasma from 46 patients with RA and 8 with osteoarthritis (OA) were measured.Those in synovial fluid (SF) from 14 patients with RA,and plasma from 12 healthy subjects were measured as controls.Results　The concentration of uPA and uPAR in SF from patients was significant higher than that in their plasma (P＜0.001,P＜0.01),and the concentration of uPA and uPAR in plasma of RA was also much higher than that in plasma of OA patients (P＜0.05,P＜0.000 1) and healthy subjects (P＜0.000 1,P＜0.001).There was no significance between plasma levels of RF＋ and RF－ in RA patients.The concentration of uPA and uPAR of plasma in RA correlated positively with CRP,RF and the number of swelling joints.Conclusion　The levels of uPA and uPAR in SF and plasma are useful parameters for monitoring disease activity of RA.These findings suggest that the uPA and uPAR genes may play an important role via proteolytic damage of the extracellular matrix during the development of RA.

Objective　To probe into roles of hCKLF-1 in the development of systemic lupus erythematosus in BXSB mice in vivo.Methods　The recombinant eukaryotic expression plasmid for hCKLF-1 (pCDI-hCKLF-1) was transferred and expressed in lupus-prone BXSB mice with the technique of muscle-mediated transgene by electric pulse;the levels of serum IgM and IgG anti-DNA antibodies as well as serum BUN and urine protein were detected.Results　hCKLF-1 expression in vivo did not make any effects on the levels of IgM and IgG anti-DNA antibodies in female and male BXSB mice,but markedly enabled abnormal elevation of urine protein in male BXSB mice in short time,suggesting its ability for accelerating glomerulonephritis.Conclusion　hCKLF-1 may be one of inflammation mediators,playing an important role in the lupus glomerulonephritis of male BXSB mice,and its target cells may be monocyte and macrophage.

Objective To immunize BALB/c mice with type 3 musearinic acetylcholine receptor polypeptide (M3RP) and to evaluate the role of M3RP in SS. Methods Four-week-old BALB/e mice were immunized with M3R polypeptide 213-228 (M3RP) on days 0, 14, 35, 56, and were re-immuniged on days 65, 84, 105, and one mouse was killed every 2 to 3 weeks. The mice of the control group were immunized with submaxillary gland homogenate, GST, and PBS. The animals were analyzed for the presence of anti-SSA,anti-SSB, RF, ANA, anti-a-fodrin and anti-M3RP in sera by immunofluorescenee or ELISA. The cytokines of IFN-γ IL-2 and IL-10 were measured with ELISA. Salivary glands were examined by H＆E staining and immunohistochemical analysis. Volume of water drinking by each groups were calculated. Results BALB/e mice immunized with M3RP and submaxillary gland homogenate developed an immune response directed against M3RP, α-fodrin and ANA, but no antibodies against SSA, SSB and RF were found. Furthermore, lym-phocytic infiltrates in the salivary glands of immunized animals were observed 50 days after first immunization of M3RP and submaxillary gland homogenate. The serum IFN-α in mice of M3RP, submaxillary gland ho-mogenate, GST and PBS was (62±6), (89±5), (30±5) and (19±6) pg/ml respectively, and IL-2 was (12.6±1.6), (19.8±0.4), (3.9±0.9), and (4.9±1.1) pg/ml respectively (P<0.05). No difference was found in the level of serum IL-10 among the four group. Expression of α-fodrin was found only in submaxillary gland in M3RP and submaxillary gland homogenate groups of mice, but not in PBS and GST controls when studied by immunohistochemical analysis. Conclusion These results suggest that BALB/c mice immunized with M3RP are reminiscent of human SS, and M3RP as an autoantigen participates the development of SS.

Objective To observe homing of mesenchymal stem cells (MSCs) in immune organs and inflammatory joints in collagen induced arthritis(CIA) rats. MethodsRats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture cell walls, and the phenotypes were assessed by flow cytometry. MSCs were labeled by PKH-26 and Brdu. Sixty-four rats were randomly divided into normal group and CIA group. Every 8 rats were sacrificed at 3, 11, 30, 42 days after transplantation of MSCs. At the end of the experiment, the specimens of thymus gland, spleen, ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. Confocal laser scanning microscope and immunohistochemical method were used to observe migration and distribution of MSCs in different organs. Independent samples group t test with SPSS 12.0 software package was used for statistical analysis. ResultsIt was found that allogenic MSCs could stay in spleen, thymus gland and joints of CIA rats for a relatively long period (42days). Forty-two days after transplantation of MSCs, the average grey scale values of spleen and thymus gland in CIA group(37.5±8.8, 29.9±5.9 respectively) were significantly higher than the normal group(16.0±2.3,13.2±4.3 respectively), the average grey scale values of ankle joints in CIA group 78±8 was significantly lower than the normal group 93±14(P<0.05). ConclusionIt has been found that MSCs can stay in the injured tissue and organs preferentially.

Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs) on the peripheral T-lymphocytes of lupus nephritis in vitro. Methods MSCs were isolated and expanded from human bone marrow cells. The purity of MSCs was identified by flow cytometry (FCM). The MSCs (4×104, 1×104, 2×103) were added into wells containing peripheral blood lymphocytes (2×105) from lupus nephritis in the presence of phytohemagglutinin [PHA). Samples were divided into the following groups: group A:T-lymphocytes alone; group B: MSCsl with T-lymphocytes(MSCsl:T=1:5); group C: MSCs2 with T-lymphocytes (MSCs2:T=1:20); group D: MSCs3 with T-lymphocytes (MSCs3:T=1:100). The proliferation of T-lymphocytes was assessed by MTT. FCM was used to analyze the apoptosis of T-lymphocytes and surface markers of CD28 and CD152. The gene expression of interferon γ (IFN-γ), interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1) were detected by real-time RT-PCR. Results In vitro, with the presence of MSCs, peripheral blood T-lymphocytes from lupus nephritis were statistically significantly decr-eased in their proliferative activities , apoptosis and CD28 expression in a dose-dependent manner. No inhibitory effects on CD152 expression of T-lymphocytes had been observed . MSCs promoted the gene expression of TGF-β1 and inhibited the gene expression of IL-10, IFN-γ. Conclusion MSCs can inhibit the proliferative activities, apoptosis and CD28 expression of peripheral blood T-lymphocytes of lupus nephritis, increase gene expression of TGF-β1 and lower the gene expression of IL-10, IFN-γ, which may play an important role in it's immunosuppressive effects on lupus nephritis.