| The First Part: Research on levels of interleukin-4, interferon-gamma, and Leukotrienes in bronchoalveolar lavage fluid from children with refractory mycoplasma pneumoniae pneumoniaObjects:To detect levels of interleukin-4, interferon-gamma, and Leukotrienes in bronchoalveolar lavage fluid(BALF) from children with refractory Mycoplasma pneumoniae pneumonia(RMPP) and explore the relationship between children RMPP and atopic status.Methods:1. Patients with RMPP (n=42) in paediatrics department of Pepple's Hospital of Hunan Province during September 2008 to March 2010 were retrospectively enrolled. The specific antibody of MP was detected by partical agglutinate method. It was taken as the diagnostic criteria of MP infection that the titer of specific antibody of MP in the second serum increasing exceed 4 times than that in the first serum. All the patients were given flexible fiberoptic bronchoscopy and their BALF were collected. We divided all the patients into two group according to whether they have eczema, allergic rhinitis, urticaria and family history of anaphylactic disease or not, atopic group(n=11) and non-atopic group(n=31). Meanwhile, re-examinated cases(n=12) who were given the operation of removing airway foreign bodies 4 weeks before were taken as control group and their paired serum, BALF were collected.2. Fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect 16S rRNA gene of MP in BALF collected by fiberoptic bronchoscopy from children with RMPP and control group.3. The differential cell count in BALF were analyzed.4. The levels of interleukin-4(IL-4), interferon-gamma(INF-γ), leukotrienes-B4(LTB4) and cysteinyl leukotriene(CysLT) in BALF were measured by enzyme linked immunosrbent assay(ELISA).5. At the same time we also detected the levels of protin in BALF.Results:1.16S rRNA gene of MP detected by FQ-PCR showed that BALF of 42 patients with RMPP were all positive. BALF were collected earlier than that titer of specific antibody of MP increased exceed 4 times.2. Compared with the control group, the total cells in BALF from children with RMPP increased significantly(P<0.05),and the percentage of neutrophil increased higher significantly(P<0.05). 3. Compared with the control group, the IL-4 and INF-y in BALF from children with RMPP increased significantly(P<0.05), the ratio of INF-γ/IL-4 decreased not significantly(P>0.05), and the IL-4 and INF-γin BALF of atopic group increased significantly(P<0.05), the ratio of INF-γ/IL-4 decreased not significantly(P>0.05), and the INF-y in BALF of non-atopic group increased significantly(P<0.05), the IL-4 and ratio of INF-γ/IL-4 was not significantly changed(P>0.05). Compared with the non-atopic group, the IL-4 and INF-y in BALF of atopic group increased not significantly(P>0.05), the ratio of INF-γ/IL-4 also decreased not significantly(P>0.05).4. Compared with the control group, the LTB4 in BALF from children with RMPP increased significantly(P<0.05), the CysLTs increased not significantly(P>0.05), and the LTB4 in BALF of atopic group increased significantly(P<0.05), the CysLTs increased not significantly(P>0.05), and the LTB4 in BALF of non-atopic group increased significantly(P<0.05), the CysLTs was not significantly changed(P>0.05). Compared with the non-atopic group, the LTB4 and CysLTs in BALF of atopic group increased significantly(P<0.05).5. The percentage of neutrophil in BALF of the patients with RMPP were positively correlated with the level of LTB4(y=0.742, P<0.01).Conclusion: 1. The method of FQ-PCR to detect MP 16S rRNA gene in BALF may be a reliable detection method in the early diaognosis of pathogen in patients with MPP.2. The total cells, especially the percentage of neutrophil in BALF from children with RMPP increased significantly,it is probably caused by the increasing of LTB4. And in atopic children the level of LTB4 are higher than non-atopic children.3. The IL-4 and INF-y in BALF from children with RMPP increased significantly, but the ratio of INF-γ/IL-4 have no difference, it supports that RMPP results in an increased proinflammatory, but there are no evidence to support a TH2-like cytokine dominating host response in RMPP.4. The CysLTs in BALF from children with RMPP have no difference, but in atopic children CysLTs have increased significantly, it suggest CysLTs may play a significant role in hypersensitiveness status, but whether CysLTs will increasing in RMPP or not is still uncertain. The Second Part: Long-term follow-up on lung function of the children who recovered from refractory mycoplasma pneumoniae pneumoniaObjects:We completed clinical questionnaire through telephone and carried out lung function test for the patients who recovered from RMPP 9-12 month later, to approach the prognosis of RMPP and explore whether there are some long-term effects on the lung function of children or not.Methods:1. The patients enrolled in the first part are all included. After they recovered from RMPP 9～12month later(token as RMPP group), we completed clinical questionnaire through telephone(n=35) and carried out lung function test(n=29) for them. We divided all the patients into two groups according to whether they have eczema, allergic rhinitis, urticaria and family history of anaphylactic disease, atopic group (n=10) and non-atopic group (telephone follow-up:n=25, lung function test:n=19). Meanwhile,healthy cases(n=12) who had no history of MP or CP infection, or wheeze were taken as control group of lung function test. To accepted lung function test, the children of control group must had no illness history in the recently 2 weeks.2. We completed clinical questionnaire by telephone after they recovered from RMPP 9-12 month later.3. We carried out lung function test for the patients who recovered from RMPP 9～12 month later. Patients younger than 4 years old accepted tidal pulmonary function test, those older than 6 years old accepted routine tests of ventilator function, and those between 4 and 6 years old accepted IOS mode test.Results:1. X-ray or CT showed that the focus of infection disappeared completely in all children after discharged 1-2 month later.8(21.74 %) had a history of cough after recovery,3 of them(8.70%) are Chronic cough.5(13.04%) had a history of wheeziness are diagnosed as asthma.1 (4.35%) had allergic purpura, and he was healed already.2. Compared with non-atopic group, the frequency of wheezinnness (asthma) in atopic group was significantly increased(P<0.05); occurrence of others have no difference (P>0.05).3. Lung function test showed:in RMPP group,9 accepted tidal pulmonary function test,2 was abnormal because of airway obstruction.11 accepted IOS mode test,1 was abnormal because of airway obstruction.9 accepted routine tests of ventilator function, all were OK. In control group, everyone was OK. There are no difference between the two groups (P>0.05).4. In lung function follow-up, there are no difference between non-atopic group and atopic group (P>0.05), and there are also no difference among different age range (P>0.05).Conclusion:1. RMPP is very severe, but an earliy and active management can still lead to a perfect result.2. We can not suggest that the RMPP will have a long-term effects on lung function of children. |
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