Object ive To explore a method to recons t ruct eccr ine sweat gland- l ike s t ructure in vitro. Methods Isolated from the normal axillary full-thickness skin donated by volunteers sweat gland epithel ial cells were cultured in vitro and were observed under inverted phase contrast microscope. These cells at the density of 2 × 105/cm2 were inoculated underneath the Matrigel (group A), on the top of the Matrigel (group B) and in the Matrigel (group C), respectively,for three-dimensional culture. The formation of eccrine sweat gland-l ike structure was observed by confocal laser scanning microscope, HE staining and immunohistochemistry staining. Results Primary epithel ial cells in the secretory portion of sweat gland were attached and spindle-shaped 24 hours after inoculation, and were under polyclonal grain-l ike growth 2-3 days thereafter. Cobblestone-l ike appearances of these cells were evident 14 days after inoculation and the confluent cells were flat and polygonal with relatively big round cell nucleus. Morphologically, subcultured cells at passage 1 were similar to the primary cells; cells at passage 2 were irregular and most of them had long pseudopodium; cells at passage 3 were star-shaped and big and had fusion with adjacent cells. For group A, tubular structure was formed 11 days after three-dimensional culture. For group B, stretched and filamentous-shaped cytoplasm was observed 8 hours after three-dimensional culture, with the formation of lumen or half-lumen structure, but no significant prol iferation was evident. For group C, cell division and prol iferation occurred 2-3 days after three-dimensional culture; the prol iferated cells were closely arranged into tubular structure with obvious lacunae in the middle, which gradually developed into irregular ball-shaped structure with the increase of neonatal cells. The laser scanning confocal microscope observation showed the formation of spherical structure in group C, with tubular structure in the center of cell mass; HE staining testified the spherical structure in group C was tubular structure. The immunohistochemistry staining demonstrated keratin 18 and carcinoembryonic antigen were positively expressed in group C, which was similar to the tubular structure of secretory portion of sweat gland. Conclusion The sweat gland epithel ial cells can be induced to form eccrine sweat gland-l ike structure through three-dimensional culture in Matrigel.
Objective To observe the clinical effect of repetitive transcranial magnetic stimulation (rTMS) combined with balloon dilatation and routine swallowing training on dysphagia caused by cricopharyngeal dysfunction (CPD) after stroke. Methods Patients with dysphagia after stroke who were hospitalized at Hubei Provincial Hospital of Integrated Chinese and Western Medicine between January 2022 and February 2023 were selected. The patients were divided into the trial group and the control group by random number table method. The control group received balloon dilatation and routine swallowing training, and the trial group received rTMS based on the treatment plan of the control group. All patients were treated for 3 weeks. The videofluoroscopic swallowing study (VFSS), Penetration Aspiration Scale (PAS), and Functional Oral Intake Scale (FOIS) were used at pre-therapy and 3 weeks after treatment to assess the improvement of swallowing function. Results A total of 49 patients were included, including 25 in the trial group and 24 in the control group. There was no statistically significant difference in age, gender, course of disease, stroke type, and swallowing function before treatment between the two groups of patients (P>0.05). After 3 weeks of treatment, the VFSS dysphagia scores (Z=−4.465, −4.327, P<0.001) of the trial group and the control group were higher than those before treatment, and the trial group was better than that in the control group (t=2.099, P=0.041). The PAS scores (Z=−4.179, −3.729, P<0.001) and FOIS scores (Z=−4.476, −4.419, P<0.001) of the trial and control groups were improved after treatment, and the improvement of the PAS score (t=−2.088, P=0.042) and FOIS score (Z=−2.134, P=0.033) in the trial group were more significant (P<0.05). No serious adverse reactions were observed in patients during the study process. Conclusion The rTMS combined with balloon dilatation and routine swallowing training can significantly improve the swallowing function of patients with dysphagia caused by CPD after stroke, and further improve its clinical efficacy, worthy of clinical application.