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Objective To investigate the effects of calcitonin gene-related peptide ( CGRP) on the inflammatory response induced by lipopolysaccharide ( LPS) of Klebsiella pneumoniae. Methods Klebsiella pneumoniae was cultured in vitro to extracted LPS. Different concentrations of LPS were used to stimulate BEAS-2B cells. The activation of human βdefensin 2 (hBD-2) in these cells was detected by immunofluo-rescence assay before and after adding different concentrations of CGRP. MTT assay and flow cytometry were respectively used to detect the proliferation and apoptosis of BEAS-2B cells after LPS stimulation with and without CGRP treatment. Neutrophil granules released in the cells after CGRP treatment were detected using neutrophil gelatinase-related apolipoprotein ( NGAL) as the marker. Results Immunofluorescence assay re-sults showed that LPS at different concentrations could significantly increase the relative expression of hBD-2 in BEAS-2B cells, which was significantly inhibit by CGRP intervention. LPS at 50 ng/ml, 100 ng/ml and 200 ng/ml had no significant effect on the activity of BEAS-2B cells, while treatment with 400 ng/ml of LPS for 24 h could significantly reduce the activity and promote the apoptosis of BEAS-2B cells. In addition, re-markedly increased cell activity and suppressed cell apoptosis were induced when BEAS-2B cells were trea-ted with 10 nmol/L of CGRP in combination with LPS. LPS at different concentrations could induce the re-lease of neutrophil-specific granules, while the LPS-induce release could be significantly inhibited by 10 nmol/L of CGRP. Conclusions CGRP could inhibit the expression of hBD-2, promote cell proliferation and reduce the degranulation of neutrophils to down-regulate the inflammatory response induced by LPS of Klebsiella pneumoniae.
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Objective To investigate the value of serum thyroid hormone (Thyroid hormone, TH) levels on prognosis in patients with treatment-resistant depression ( Refractory depression, RD) .Methods 108 cases of RD patients collected in Hangzhou First People's hospital from March 2014 to February 2016 were divided into observation group and control group according to the method of random numbers, and each had 54 patients.Patients in the observation group were given antidepressant treatment program and the control group a placebo.Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA) score changes, and TSH, T3, T4, FT3, FT4 levels of two groups pre-and post-treatment were compared.Results After treatment, the HAMD and HAMA scores in observation group were (14.4 ±2.5) and (15.2 ±2.7) significantly lower than that in control group, which were (25.6 ±5.2) and (25.9 ±4.8),separately, the differences were significant(P<0.05);the TSH, T3, T4, FT3, FT4 levels in observation group post-treatment were (4.54 ±0.68) mIU/L, (1.21 ±0.56) nmol /L, (55.4 ±6.1) nmol/L, (3.16 ±0.42) pmol/L and (8.53 ±0.62) pmol/L, TSH decreased significantly, and T3, T4, FT3 , FT4 were significantly increased than pre-treatment(P<0.05); the indexes in control group were (5.16 ±0.62) mIU /L, (0.91 ±0.42) nmol/L, (51.9 ±3.2) nmol /L, (2.82 ±0.40) pmol/L and (7.76 ±0.64) pmol/L (P<0.05), the differences with observation group were significant ( P <0.05 ) .Conclusion The severity of the symptoms of patients is closely related with TH RD level, antidepressant therapy can improve TH, thereby improving the patient's symptoms, so it is important to pay attention to TH levels in the treatment process of RD.
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Objective To prepare a novel monoclonal antibody (mAb) that specifically against Mycobacterium tuberculosis culture filtrate protein 10 (CFP-10).Methods The BALB/c mice were immunized by a peptide with 14 amino acids (aa residues 53 to 66) of CFP-10,and then the splenocytes of mice were fused with myeloma cell line SP2/0.The resultant fused cells were subjected to screening culture,enzyme linked immunosorbent assay (ELISA) assay and subcloning by limited dilution to establish hybridoma cell lines of stable secreting anti-the peptide of CFP-10 antibody.The antibody was purified,and its isotypes were analyzed.Then,the antibody was further evaluated by Western blotting,immunoprecipitation and ELISA in 38 culture supernatant samples of Mycobacterium tuberculosis,20 culture supernatant samples of non-Mycobacterium tuberculosis,32 samples of tuberculous pleural effusion,24 samples of non-tuberculous pleural effusion,and 20 serum samples from healthy controls.Results The isotype of the mAb against the specific peptide of CFP-10 was an IgG1 with κ chain,and it was applicable for Western blotting and immunoprecipitation analysis.ELISA quantitative test showed that the sensitivity and specificity for diagnosis of Mycobacterium tuberculosis were 78.6％ (55/70) and 92.2％ (59/64),respectively.Conclusion The mAb generated against the specific peptide of CFP-10 is high in sensitivity and specificity,and it might be used in the early diagnosis of tuberculosis.
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Serum concentrations of TSH, total and free thyroxine, and total and free triiodothyronine in 592 healthy children aged 2-6 years were determined by means of ADVIA Centaur. The normal reference ranges were calculated. The reference ranges of TSH, T3, T4, FT3, and FT4 were 1.09-4.60 mIU/L, 1.17-1.95 μg/L,57.43-99.60 μg/L, 5.35-7.50 pmol/L, 13.96-19.70 pmol/L, respectively. There was no significant difference in the interval of thyroid hormones between males and females as well as among different age groups.
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Objective A recombinant adenoviral vector containing mIL-18BP and mIL-4 fusion gene(AdmIL-18BP/mIL-4) was constructed and used to investigate the role of mIL-18BP and mIL-4 in medula-ring the expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthetase (iNOS) and their inducing products PGE2, NO in murine collagen-induced arthritis. Methods Male DBA-1/BOM mice were used in this study. Mice with CIA were intra-articularly injected with 107 pfu/6μl of AdmIL-18BP/mIL-4.Intra-articular injections of AdLacZ or PBS were used as controls. The mRNA expression of COX-2, iNOS in synovial tissue was analyzed by semi-quantitative RT-PCR. Expression of COX-2 and iNOS protein was estimated by Western blot method. The production of PGE2 and NO in synovia was detected by competitive ELISA and enzyme reduction of nitrate. Results The expression of COX-2, iNOS mRNA in routine synovial tissue of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group (0.15 vs 0.42,P<0.01 ; 0.05 vs 0.77, P<0.01) and PBS group (0.15 vs 0.65, P<0.01; 0.05 vs 0.64, P<0.01 ). And the protein expression of COX-2, iNOS from AdmIL-18BP/mIL-4 treatment group was also obviously lower than that of AdLacZ group (0.08 vs 0.92, P<0.01; 0.11 vs 1.00, P<0.01) and PBS group (0.08 vs 0.77, P<0.01; 0.11 vs 0.84, P<0.01 ). The PGE2 and NO production in synovia of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group [(0.68x0.06) vs (2.58±0.21)ng/mL, P<0.01; (23.4+2.5) vs (60.0±11.3)μmol/L, P<0.01 ] and PBS group [(0.68±0.06) vs (2.57±0.20)ng/mL, P<0.01; (23.4+2.5) vs (60.3±13.4)μmol/L, P<0.01]. Conclusion These data indicat that local over-expre-ssion of mIL-18BP and mIL-4 can down-regulate COX-2, iNOS and their induced product PGE2, NO in CIA mice. The combination treatment with mIL-18BP and mIL-4 is a promising therapeutic target for RA.
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Objective To study the function of immunocytes in synovial fluid and the mechanisms of immune function disorder in patients with rheumatoid arthritis(RA).Methods The synovial fluid of 20 patients with RA and 10 healthy amputees because of traumatism in the affiliated hospital of Zhejiang university were collected in this syudy.Monoclonal antibodies against CD28,CD154(CD40L),CD80,CD86,CD40,CD69,CD25,HLA-DR,CD71 were used for flow cytometry and the expressions of costimulatory molecule CD28,CD154,CD80,CD86,CD40 on immunocytes and lymphocyte activation markers CD69,CD25,HLA-DR,CD71 in synovial fliud were studied in these patients.We also used enzyme linked immunosorbent assay(ELISA) to detect the cytokines level of Th1 cell including interleukine 2(IL-2),interferon-gamma(IFN-?) and those of Th2 cell including interleukine 6(IL-6),interleukine 10(IL-10) in the synovial fliud.Results Compared with control group,the expressions of CD80[(2.09?0.59)%vs(0.67?0.57)%,P