Efficacy of rituximab in maintenance therapy for antineutrophil cytoplasmic antibody-associated vas-culitis / 中华风湿病学杂志

Objective:To evaluate the efficacy and safety of rituximab(RTX) as remission-mainten-ance therapy in antineutrophil cytoplasmic antibody(ANCA) associated vasculitis(AAV).Methods:Patients with AAV, including granulomatosis with polyangiitis (GPA) and microscopic polyangiitis (MPA), treated with rituximab (RTX) in Peking Union Medical College Hospital during September 2005 to June 2021 were included into this study. Clinical data, relapse rate, time of first relapse and adverse events were collected and analyzed. The cumulative relapse rate was calculated by Kaplan-Meier, t test, and Man-Whithey U test and chi-square were used to compare differences between two groups. Results:① Thirty-nine AAV patients were enrolled, including 36 GPA and 3 MPA. During the 20(3, 104) months follow-up, 59.0%(23/39) patients had suffered relapses. The time for first relapse was 11(3, 42) months after remission. ② There were no difference in the relapse rate [60.0%(18/30) vs 55.6%(5/9), χ2=0.06, P=1.000), the time of first relapse [15(3, 42) vs 10(9, 30), Z=0.45, P=0.678], CD19 + B [23.5 (5, 148) cell/μl vs 3(2, 15) cell/μl, Z=0.57, P=0.605] and serum IgG [7.09(5.13, 13.90) g/L vs 9.72(5.32, 12.0) g/L, Z=0.36, P=0.770] between standard dose and low-dose groups. The rate of major relapse-free was significantly less in patients treated with standard dose than patients with reduced dose of RTX {87.1%[95% CI(73.4%, 100.8R%)] vs 64.3%[95% CI(23.1%, 105.4%)], χ2=7.59, P=0.006}. ③ There were no difference in relapse rate [50.0%(3/6) vs 60.6%(20/33), χ2=0.24, P=0.674], time of first relapse [23(6, 25) vs 11(3, 42), Z=0.05, P=0.982], CD19 + B[35(15, 50) cell/μl vs 10(0, 148) cell/μl, Z=0.95, P=0.382] and serum IgG[6.70(5.91, 7.49) g/L vs 7.69(3.78, 13.90) g/L, Z=0.48, P=0.700] between the fixed interval dosage and the on-demand dosage groups. There was no difference in the rate of major relapse-free between the two groups (100% vs 77.8%, χ2=1.79, P=0.181). ④ The incidence of infusion reaction was 5.1%(2/39) and infection was 20.5%(8/39). Serum IgG level was 4.37(3.78, 13.4) g/L at infection. There was no difference in safety between the standard and low-dose groups or between fixed interval and on-demand dosage groups ( P>0.05). Conclusion:There is no significant difference in relapse rate bet-ween the standard RTX dose and low-dose RTX induction therapy group, but the major relapse rate is sign-ificantly reduced in the standard dose RTX therapy. The relapse rate of fixed intervals dosage group is similar to that of on-demand dosage group. The safety profile of the standard dose and low-dose induction therapy groups or fixed intervals and on-demand dosage groups is similiar.

Effects of fibrobl ast growth factor 4 on the proliferation of fibroblast-like synoviocytes in rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression of fibroblast growth factor 4 (FGF4) in serum of active rheumatoid arthritis (RA) and its role in RA synoviocyte proliferation. Methods The serum level of FGF4 were detected by protein arrays in 20 patients with RA, and 20 age and gender matched healthy controls. FLSs were isolated from RA synovium,and were co-cultured with recombinant human FGF4 (rhFGF4). Cell proliferation was quantified by Cell Counting Kit-8 assay and cell cycle distribution was evaluated by flow-cytometry. The protein levels of cyclin D1, phospho-Akt (p-Akt) and phospho-p38 (p-p38) were measured by western blot. Results The serum expression of FGF4 in RA group was higher than that in control group (P=0.041). After being treated with different concentrations of rhFGF4 (12.5, 25, 50, and 100 ng/ml), RA-FLS showed significant increase in cell proliferation, with different rates of [(121 ±8)％], [(126 ±12)％], [(129 ± 12)％], a nd [(134 ±14)％] respectively, comparing with that of the controls [(100 ±0)％, (P12.5=0.049, P25=0.009, P50=0.004, P100=0.001).]. Among them, the percentage of G2/M+S phase cells were [(12.6±3.6)％], [(15.3±4.5)％], [(17.1±5.1)％], [(19.6±4.1)％] respectively, and except the lowest rhFGF4 concentration treatment group of 12.5 ng/ml, G2/M+S phase cells in other groups was significantly increased compared with the controls [(5.4±2.4)％] (P12.5=0.159, P25=0.042, P50=0.018, P100=0.005). And the protein expression of cyclin D1 was up-regulated after being treated with 50 ng/ml and 100 ng/ml rhFGF4 (P50=0.035, P100=0.027). FGF4 transiently increased the expression of p-Akt and p-p38 protein at the concentration of 50 ng/ml. Comparisons of data between groups were performed by independent sample Student's t-test. Statistical significant differences among groups were tested by one-way analysis of variance (ANOVA) or the Kruskal-Wallis test. The Dunnett's t-test was used for multiple comparisons. A P-value of <0.05 was considered statistically significant. Conclusion Our results suggest that FGF4 is highly expressed in the serum of active RA patients. FGF4 may promote the proliferation of RA-FLS via modulating PI3K/Akt and p38-MAPK signaling pathways, which subsequently contributs to synovial hyperplasia.

Human IgG inhibits IgA1 protease-dependent adherence of Haemophilus influenza strains to human lung epithelial cells

    Keywords: Cellular adherence, iga gene; IgA1 protease subtypes, IgG inhibition, Neisseria meningitides, typableand nontypable Haemophilus influenzae (THi and NTHi)Background: IgA1 protease may enhance the bacterial infection in human beings. However, the molecular mechanism of bacterial adherence to eukaryotic cells is unclear.Objective: Reveal the mechanisms of IgA1 protease-dependent and non-protease bacterial adherence toeukaryotic cells.Method: Type I and type II IgA1 proteases from iga genes (GenBank DQ683355 for NTHi465, DQ683356 for NTHi500 and DQ683357 for Nm430) were cloned, expressed, and purified. Cellular assays for adherence of IgA1 protease-producing and -non-producing and typable and nontypable strains of H. influenzae to human lung carcinoma cells (A549) were carried out in the presence of human antibodies.Results: Adherence of protease-producing strains and non-producing strains to human epithelial cells was significantly dependent on the enzyme activity. In addition, human IgG was an inhibitor to IgA1 proteasedependent adherence of H. influenzae strains to human cells. However, IgA1 antibodies were irrelevant to IgA1 protease-dependent adherence.Conclusion: IgA1 protease was required for adherence of pathogenic bacteria to human epithelial cells in IgA1 protease-producing bacteria, and human IgG inhibits the adherence, but not for IgA1 protease non-producing bacteria.

Infection of Schistosomiasis japanicum is likely to enhance proliferation and migration of human breast cancer cells：mechanism of action of differential expression of MMP2 and MMP9

Objective: To study whether the infection of Schistosomiasis japanicum (S. japanicum) is related to enhanced proliferation and migration of cancer cells, and the molecular mechanism pertains to cancer cell metastasis in human host. Methods: The gene of S. japanicum glutathione transferase (sjGST) cloned from S. japanicum was expressed, purified and applied in a series of assays to explore the effect of sjGST on proliferation and migration of MDA-MB-435S, and the expression of MMP2 and MMP9. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S was also carried out. Results: Results showed that sjGST enhanced proliferation and migration in human breast cancer cell MDA-MB-435S signifycantly at 50-200 nM, but did not enhance them in human lung cancer cell A549. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S and A549 showed that GST was readily bound to the breast cancer cells, but showed almost no binding to human lung cancer cells. The assays for gelatinase activity showed that both MMP2 and MMP9 activities were increased significantly in the presence of sjGST (50-200 nM) in MDA-MB-435S, but they were not significant in A549. Conclusions: Our current results show strongly that S. japanicum GST binds to MDA-MB-435S probably via its receptor, and enhances proliferation and migration of the cancer cells by up-regulatory expression of MMP2 and MMP9.

Infection of Schistosomiasis japanicum is likely to enhance proliferation and migration of human breast cancer cells: mechanism of action of differential expression of MMP2 and MMP9

<p><b>OBJECTIVE</b>To study whether the infection of Schistosomiasis japanicum (S. japanicum) is related to enhanced proliferation and migration of cancer cells, and the molecular mechanism pertains to cancer cell metastasis in human host.</p><p><b>METHODS</b>The gene of S. japanicum glutathione transferase (sjGST) cloned from S. japanicum was expressed, purified and applied in a series of assays to explore the effect of sjGST on proliferation and migration of MDA-MB-435S, and the expression of MMP2 and MMP9. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S was also carried out.</p><p><b>RESULTS</b>Results showed that sjGST enhanced proliferation and migration in human breast cancer cell MDA-MB-435S signifycantly at 50-200 nM, but did not enhance them in human lung cancer cell A549. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S and A549 showed that GST was readily bound to the breast cancer cells, but showed almost no binding to human lung cancer cells. The assays for gelatinase activity showed that both MMP2 and MMP9 activities were increased significantly in the presence of sjGST (50-200 nM) in MDA-MB-435S, but they were not significant in A549.</p><p><b>CONCLUSIONS</b>Our current results show strongly that S. japanicum GST binds to MDA-MB-435S probably via its receptor, and enhances proliferation and migration of the cancer cells by up-regulatory expression of MMP2 and MMP9.</p>