慢性阻塞性肺部疾病(COPD)是全球性高发病率、高死亡率以及高卫生保健费用的重要疾病之一。2001年COPD是发达国家第5位的致死原因，占总死亡数的3．8％；在发展中国家则为第6位致死原因，占总死亡数的4．9％[1]。随着全球人口的老龄化，COPD负担将逐年增加。因此，在未来数年内我们必须共同面对挑战，实施有成本效益的防治策略，以遏制这一疾病及其耗费。
支气管哮喘是由嗜酸粒细胞、 肥大细胞和T淋巴细胞等多种细胞和细胞组分参与的气道慢性炎症性疾患，主要病理特点为上皮层大量的嗜酸粒细胞浸润及以上皮下纤维化、平滑肌增生、胶原蛋白沉积为主的气道重塑。哮喘的发病机制较为复杂，而炎症因子表达异常在哮喘的发病中发挥主要作用[1-3]。哮喘是一种全球范围内的常见病、多发病，我国约有1000万以上哮喘患者，而目前对于哮喘尚缺乏有效的根治方法。 间充质干细胞（mesenchymal stem cells，MSC）是具有强大的增殖能力和多向分化潜能的成体干细胞，同时具有免疫调节作用，它能通过免疫调节作用改善多种免疫相关性疾病的病情，而既往MSC在呼吸系统疾病中的研究主要集中在急性肺损伤，在哮喘当中的研究甚少。对于哮喘这一类以炎症因子表达异常为主的变态反应性疾病，MSC是否可以用于哮喘的治疗，值得我们进一步探讨。
ObjectiveTo study immunodepression effect of bone marrow-derived mesenchymal stem cell (BMSC) on acute asthmatic airway inflammation by galectin-1 (gal-1) in vivo.MethodsEighty-five female BALB/c mice were equally randomized into normal control group, asthmatic group, BMSC treatment group, gal-1 treatment group and BMSC and gal-1 inhibitor group. Ovalbumin (OVA) was used to establish acute asthmatic model. Total cell number and differential cell analysis in each group in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, hematoxylin-eosin and periodic-acid Schiff staining was used to compare airway inflammation among five groups. Measurement of cytokines, including interleukin (IL) -4, IL-5 and gal-1 in BALF and OVA specific IgE (OVA-IgE) in serum were evaluated by enzyme linked immunosorbent assay. Moreover, dendritic cell (DC) in lung tissue was sorted by immunomagnetic beads and its MAPK signal pathway was analyzed by western blotting among five groups.ResultsAccumulation of inflammation cells, particularly eosinophils around airway and in BALF was evident in asthmatic mouse model, meanwhile hyperplasia of Goblet cell was also obvious in asthmatic group. BMSC engraftment or gal-1 infusion significantly reduced airway inflammation and hyperplasia of Goblet cell and the number of inflammation cells in BALF, especially eosinophils attenuated dramatically. However, there was no effect on airway inflammation and hyperplasia of Goblet Cell by simultaneous infusion BMSC engraftment and gal-1 inhibitor. Compared to normal control group, the level of IL-4, IL-5 in BALF and OVA-IgE in serum was increased remarkably in asthmatic group, but the level of gal-1 reduced obviously. Moreover, infusion of BMSC or gal-1 could mitigate the level of IL-4, IL-5 in BALF and OVA-IgE in serum and increase the level of gal-1 in asthmatic mouse. However, infusion with both BMSC and gal-1 inhibitor exerted no effect on cytokine and OVA-IgE in asthmatic mouse. DC was sorted by immunomagnetic beads and western blotting was used to detect the expression of MAPK signal pathway among five groups. The expression of ERK phosphorylation in asthmatic group was much lower than that in normal control group. On the contrary, the expression of p38 phosphorylation was much higher than that in normal control group. BMSC engraftment or gal-1 infusion significantly activated the ERK pathway and inhibited the p38 MARP pathway on asthmatic mouse DC. Nevertheless, the expression of ERK phosphorylation and p38 phosphorylation for group with BMSC and gal-1 inhibitor infusion was between the level of asthmatic group and normal control group.ConclusionsBMSC infusion alleviates airway inflammation in asthmatic mouse, especially weakens eosinophils infiltration, and the underlying mechanism might be protective effect of gal-1 secreted by BMSC which plays a role in lung tissue DC and regulates the DC expression of MAPK signal pathway.
【Abstract】 Objective To investigate the effect of allogeneic bone marrow-derived mesenchymal stem cells ( BMSCs) transplantation on the airway inflammation and airway remodeling in chronic asthmatic mice. Methods Forty female BALB/c mice were equally randomized into four groups, ie. a normal control group, a BMSCs control group, an asthma model group, and a BMSCs transplantation group. BMSCs were generated from male donor mice, then the mice in the asthma model group and the BMSCs transplantation group were sensitized and challenged with OVA to establish chronic asthmatic mice model. Hematoxylin and eosin staining and Alcian blue-periodic acid-Schiff staining were used to analyze the effects on airway inflammation and airway remodeling after BMSC engraftment. The number of CD4 + CD25 + regulatory T cells in spleen was detected by flow cytometry. Results In lungs of the asthmamodel group, there were intensive inflammatory cells infiltration around airway and blood vessels, goblet cell proliferation, epithelial desquamation, patchy airway occlusion by hyperviscous mucus, and hypertrophy of airway smooth muscle.Airway inflammation and airway remodeling were significantly relieved in the BMSCs transplantation group.There was no obvious inflammatory cells infiltration in the airway and airway remodeling both in the normal control group and the BMSCs control group. The number of CD4 + CD25 + regulatory T cells in spleensignificantly decreased in the asthma model group compared with the two control groups ( P lt; 0. 05) , and significantly increased in the BMSCs transplantation group compared with the asthma model group ( P lt;0. 05) . There was no significant difference in the number of CD4 + CD25 + regulatory T cells in spleen betweenthe control groups and the BMSCs transplantation group. Conclusion BMSCs engraftment can up-regulate CD4 + CD25 + regulatory T cells and relieve airway inflammation and airway remodeling in asthmatic mice.
ObjectiveTo establish a simple and stable model of benign tracheal stenosis in SD rats by nylon brush scraping induced mechanical injury, and to observe the pathological changes of tracheal tissue at different time points after modeling.MethodsTwenty SD rats were divided into sham operation group (10 rats) and stenosis model group (10 rats) by random number method. Symptoms and survival conditions were observed, tracheal tissues were obtained, granulation tissue proliferation was observed, and stenosis indexes were measured and compared. Another fifteen rats were sacrificed at different time points (days 0, 2, 4, 6, and 8) after modeling. Tracheal tissues were obtained, HE staining and Masson staining were performed to observe pathological changes with time.ResultsThe survival rate of the sham operation group was 100% on the 8th day after operation, and the survival rate was 0% on the 8th day after operation in the stenosis model group. The difference in survival condition between the two groups was statistically significant (P=0.000 1) by Log-rank test. The stenosis index in the sham operation group was (6.12±1.78)%, and in the stenosis model group was (60.28±12.56)%. The difference in the stenosis between the two groups was statistically significant (P<0.000 01). HE staining results showed that the tracheal lumen was unobstructed and no granulation tissue hyperplasia or stenosis was found in the sham operation group. The epithelial mucosa was intact and smooth, and the cilia structure was clearly visible. It was a pseudo-stratified ciliated columnar epithelium, which was consistent with the characteristics of normal airway mucosa. While in stenosis model group, the lumen was significantly narrowed, and the stenosis was mainly caused by granulation tissue hyperplasia. No epithelial structure was observed, or epithelial structure was extremely abnormal. Masson staining showed that the fibroblasts in the injured site increased first and then decreased, and the collagenous fiber (blue) in the injured site gradually increased with time.ConclusionsA model of benign tracheal stenosis in rats can be successfully established by nylon brush scraping induced mechanical injury. The modeling method is simple, controllable and reproducible. The model can be widely used in the investigation of pathogenic mechanism for benign airway stenosis and efficacy exploration of new treatment.
ObjectiveTo explore the efficacy of community-acquired pneumonia (CAP) by tracheoscopy intervention altimeter and analyze and compare its financial burden.MethodsRetrospective analysis of 419 hospitalized patients with CAP was carried in respiratory medicine department of four hospitals from July 1, 2017 to August 31, 2018 (Changhai Hospital, Shanghai First People’s Hospital, Baoshan Branch of Shanghai First People’s Hospital, and Baoshan Integrated Traditional Chinese and Western Medicine Hospital). According to the time of tracheoscopy intervention treatment, they were divided into 3 groups: 127 patients treated with tracheoscopy intervention during the initial treatment period (within 72 h after obtaining imaging diagnosis) were included in an early intervention group, 158 patients treated with tracheoscopy intervention 72 h after obtaining imaging diagnosis were included in a medium-term intervention group, and 134 patients treated without tracheoscopy intervention were included in a non-intervention group. The total efficiency of treatment, improvement of clinical symptoms, imaging absorption, serum inflammation index level, sputum culture positive rate, change rate, efficiency after drug change, hospital stay and hospitalization cost were compared among three groups.ResultsThe total efficiency of treatment in the early intervention group was higher than that of the medium-term intervention group and the non-intervention group, with statistically significant difference (P<0.05), and the time of normality of body temperature, the time of disappearance of strong sputum and cough in the early intervention group, the absorption time of chest X-rays were shorter than that of the medium-term intervention group and the non-intervention group, and the difference was statistically significant (P<0.05); peripheral blood hemoglobin, serum calcitonin and hypersensitive C reactive protein levels were lower than those in the medium-term intervention group and the non-intervention group, with statistically significant differences (P<0.05), and the sputum-positive and drug-change rates in the early intervention group and the medium-term intervention group were higher than those in the non-intervention group, and the difference was statistically significant (P<0.05); the duration of hospital stay in the early intervention group was shorter than that of the medium-term intervention group and the non-intervention group, and the cost of hospitalization was less than that of the medium-term intervention group and the non-intervention group, and the difference was statistically significant (P<0.05).ConclusionTracheoscopy intervention treatment in the initial period of CAP not only significantly improves the efficacy, but also significantly reduces treatment costs and length of hospitalization, hence it is worth clinical promotion.
ObjectiveTo introduce implantation methods of different types of congenital heart disease occluder for the treatment of bronchopleural fistula (BPF) and its preliminary efficacy.MethodsThree patients who diagnosed with BPF and treated by congenital heart disease occluder were reviewed. The clinical data was analyzed after comprehensively reviewing of relevant literature.ResultsAll the three patients were treated with postoperative BPF and empyema. The diameter of the fistula ranged from 3 to 8 mm. We used occlusive devices for congenital heart diseases such as atrial septum (ASD), ventricular septum (VSD) defect or patent ductus arteriosus (PDA), respectively. After treatment, all three patients were cured of BPF and empyema caused by BPF in a short time, and the thoracic drainage tube was successfully removed. During the follow-up period from 7 to 25 months, no significant long-term complications were observed.ConclusionThe use of ASD, VSD and PDA occluder for the treatment of BPF with a fistula more than 3 mm is effective and safe.
Objective To invesitgate the clinical characteristics, radiology, diagnosis and treatment of pulmonary cryptococcosis ( PC) . Methods The patients with PC diagnosed form January 2000 to January 2009 from three hospitals of Shanghai and Nanjing were retrospectively analyzed. Results A total of 34 patients were diagnosed, with 24 males and 10 females, and an average age of ( 40. 6 ±13. 5) years old ( ranged from 3 to 72 years) . Twelve patients had underlying diseases and 28 patients had symptoms. The main symptoms were pyrexia ( 20 cases) , cough and expectoration ( 22 cases) , chest pain ( 8 cases) , chest tightness ( 5 cases) , and hemoptysis ( 4 cases) . Seven cases were diagnosed as systemic pulmonary cryptococcosis, in which 3 cases were complicated with cryptococcal meningitis ( CM) , and 3 cases with CM and cryptococcal septicemia, and 1 cases with third dorsal vertebra Busse-Buschke disease. Radiologic manifestations showed multiformand nonspecific lesion such as nodus or nodules in 17 cases, pneumonia in 10 cases, mixed appearance in 6 cases, and diffused military nodes in 1 case. The diagnosis was confirmed by pathological study in 27 cases, including 12 cases by thoracotomy, 10 cases by percutaneous lung biopsy, 1 cases by thoracic vertebra biopsy, and 4 cases by bronchoscope. Ten cases were confirmed with culture positive or smear positive. Six cases were treated by surgery alone, 21 cases by antimycotic drug therapy alone, and 6 cases by drug therapy after surgery. One case quitted after the diagnosis. The duration of treatment varied from 2 weeks to 2 years. One case died in the hospital, 25 cases recovered after discharge, and 8 cases were lost to follow-up. Conclusions PC is likely to be misdiagnosed due to atypical clinical and image manifestations. The diagnosis is always comfirmed on the pathological and microbiological study.
ObjectiveTo establish a method of air-liquid interface culture and ciliary beat frequency measurement of mouse tracheal-bronchial epithelial cells to simulate the physiological function of airway epithelium.MethodsBALB/c mouse tracheal-bronchial epithelial cells were obtained by digestion with 1 mg/mL protease in cold temperature overnight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. After removing fibroblasts by differential velocity adhesion method, the cells were cultured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different culture media.ResultsCell numbers obtained by cold protease overnight digestion for 12 h, 14 h and 16 h were (1.78±0.33)×105, (1.93±0.26)×105 and (2.01±0.28)×105, respectively. Cell viability by trypan blue staining were (96.86±0.25)%, (94.73±1.63)% and (86.87±5.95)%, respectively. Cells were 100% confluent in Transwell chamber after 1-week proliferation, and the ciliary beat frequency was observed under microscope after 2 - 3 weeks of air-liquid interface culture. The cilia structure was confirmed by hematoxylin-eosin staining, electron microscopy and immunofluorescence. Ciliary beat frequency of the cells obtained by this method was consistent with that of mouse trachea in vivo, which further demonstrated its capacity in simulating the physiological function of airway epithelium. ConclusionsThe separation and air-liquid interface culture system as well as the ciliary beat frequency measurement method established in this experiment is simple, stable, efficient and reliable, which establishes a substantial foundation for exploring the pathogenesis and treatment mechanism of airway diseases. It can also provide reference for the culture of epithelium in the airway of other species and/or other organs.