ObjectiveTo investigate the effect of graphene oxide (GO)-carboxymethyl chitosan (CMC) hydrogel loaded with interleukin 4 (IL-4) and bone morphogenetic protein 2 (BMP-2) on macrophages M2 type differentiation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).MethodsGO solution was mixed with CMC, then the phosphate buffered saline (PBS), IL-4, BMP-2, or IL-4+BMP-2 were added to prepare different GO-CMC hydrogel scaffolds with or without different cytokines under crosslinking agents. The characteristics of pure GO-CMC hydrogel were characterized by gross observation, scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FTIR), and the CMC hydrogel was used as control. The sustained release of GO-CMC hydrogels with different cytokines was also tested. Macrophages were isolated and cultured from female Sprague Dawley rats aged 4-5 weeks, and then cultured with GO-CMC hydrogels with and without different cytokines, respectively. CD206 immunofluorescence staining was used to detect the differentiation of macrophages after 24 hours. The 3rd generation of rats BMSCs were cultured with GO-CMC hydrogels with and without different cytokines respectively for osteogenic induction. The early osteogenesis was observed by alkaline phosphatase (ALP) staining after 10 days, and the late osteogenesis was observed by alizarin red staining after 21 days.ResultsGenerally, GO-CMC hydrogel was brown and translucent. SEM showed that the pore diameter and wall thickness of GO-CMC hydrogel were similar to that of CMC hydrogel, but the inner wall roughness increased. FTIR test showed that CMC polymerized to form hydrogel. In vitro, the sustained release experiments showed that the properties of GO-CMC hydrogels loaded with different cytokines were similar. CD206 immunofluorescence detection showed that GO-CMC hydrogels could induce macrophages differentiation into M2-type. ALP and alizarin red staining showed that GO-CMC hydrogels could induce BMSCs osteogenic differentiation, in which GO-CMC hydrogel loaded with IL-4+BMP-2 showed the most significant effect (P<0.05).ConclusionThe GO-CMC hydrogel loaded with IL-4 and BMP-2 can induce macrophages differentiation into M2-type and enhance the ability of BMSCs with osteogenic differentiation in vitro, which provide a new strategy for bone defect repair and immune regulation.
Objective To analyze the clinical data of patients with Tropheryma whipplei pneumonia, and summarize the clinical characteristics, diagnosis, and treatment methods of Tropheryma whipplei pneumonia. Methods The data of Tropheryma whipplei pneumonia patients from three hospitals in Hunan Province between January 1, 2021 and October 1, 2022 were retrospectively collected. The clinical symptoms, laboratory examination, metagenomics next-generation sequencing (mNGS), CT imaging features, diagnosis and treatments of the included patients were analyzed. Results A total of 4 patients were included. Among them, there were 2 males and 2 females. The main manifestations were cough, expectoration, fever, and shortness of breath. There were 2 cases of diffuse ground glass opacity in both lungs, 1 case of pulmonary nodule, 1 case of pulmonary cavity, 1 case of pleural disease, 2 cases of pulmonary exudative lesions, and 1 case of mediastinal lymphadenectasis. The mNGS results showed that Tropheryma whipplei was detected in all 4 patients, and the median number of serial number (lower quartile, upper quartile) was 1 528 (1 480, 1 576). After anti infection treatment, 3 cases were treated effectively, and 1 case had poor treatment effect. Conclusions mNGS is an effective method to diagnose Tropheryma whipplei pneumonia. The measurement of serum lactate dehydrogenase level is helpful to evaluate the disease and determine the prognosis. Piperacillin tazobactam, meropenem and doxycycline are effective for this disease, while moxifloxacin and trimethoprim / sulfamethoxazole are not recommended because they may be naturally resistant. Without active etiological treatment, the disease may persist in migration and lead to extrapulmonary involvement.