Objective To investigate the effects of extracellular signal regulated kinase ( ERK)signaling pathway on cell cycle of airway smooth muscle cells( ASMCs) in asthmatic rats. Methods Thirty Wistar rats were randomly assigned to a control group and an asthma group( 15 rats in each group) . Asthma model was established by ovalbumim sensitization and challenge. ASMC were isolated and cultured in vitro. The ASMCs from the asthmatic rats were treated with ERK activator epidermal growth factor ( EGF)and inhibitor PD98059, respectively. The expressions of cyclin D1 and CDK2 in ASMCs were detected by immunocytochemical staining. The expressions of ERK1 /2 and p-ERK1 /2 protein were observed by western blotting for measurement of ERK activation rate. Results Compared with the control group[ 54. 17 ±6. 11,61. 04 ±4. 09, ( 49. 91 ±3. 26) % , respectively] , the expressions of cyclin D1 protein and CDK2 protein,and the rate of ERK activation of ASMCs from the asthmatic rats significantly increased[ 76. 15 ±4. 88,92. 30 ±7. 95, ( 82. 37 ±5. 78) % , respectively] ( P lt; 0. 05) . Furthermore, compared with those before treatment, the expression of cyclin D1 and CDK2, and the rate of ERK activation of ASMCs significantly decreased after treatment with PD98059 [ 58. 78 ±4. 60, 69. 15 ±5. 83, ( 54. 01 ±4. 12) % , respectively]( P lt; 0. 05) , and significantly increased after treatment with EGF[ 119. 28 ±8. 14, 134. 77 ±9. 26, ( 91. 57 ±5. 32) %, respectively] ( P lt;0. 05) . Conclusion ERK1/ 2 participates in proliferation regulation of ASMCs in asthma by enhancing the expressions of cyclin D1 and CDK2, which promotes quiescent cells into S phase.
【Abstract】 Objective To investigate the biocharacteristics of c-kit+ human amniotic fluid-derived mesenchymal stemcells (HAFMSCs) and its capacity to differentiate into cardiomyocytes in vitro. Methods Fifty samples of human amnioticfluid were obtained from amniocenteses or voluntary termination of pregnancy and were expanded in vitro. c-kit+ HAFMSCs were sorted by flow cytometry and were recultured in the same media. c-kit+ HAFMSCs were amplified and identified, then exposed to osteogenic , adi pogenic, and myogenic media. The flow cytometry, and immunocytochemistry were used for identifying the cell phenotype, von Kossa staining for osteogenic differentiation, oil red O staining for adi pogenic differentiation, and real-time fluorescent quantitative PCR for the expressions of NKx2.5, Tbx5, GATA-4, and α-MHC genes. Results After the selection procedure, the percentage of c-kit+ HAFMSCs was 3.07% ± 1.03% of the total adherent cells. The cells expressed MSCs markers (CD29, CD44, CD73, CD90, CD105), and did not express hematopoietic stem cells markers (CD34, CD45). The cells were positive for human leukocyte antigen (HLA)-ABC, and negative for HLA-DR. They also expressed Oct-4, which characterized the undifferentiated stem cell state. The growth curves of c-kit+ HAMFSCs at passages 5, 10, and 15 were similar. Li pid droplet was observed by oil red O staining and calcium deposition by von Kossa staining in the cells at 21 days after adi pogenic and osteogenic induction. The myocardium special gene expressions of Tbx5, Nkx2.5, GATA-4, and α-MHC were significantly ber after myogenic induction than those before myogenic induction (P lt; 0.05). Conclusion Selected c-kit+ HAFMSCs by flow cytometry is a group of pure MSCs, which has potential to differentiate into cardiomyocytes and can be used as seeding cells for myocardium regeneration treatment.
【Abstract】 Objective To isolate and culture human amniotic fluid-derived mesenchymal stem cells (HAFMSCs),to investigate a better cryopreservation protocol of HAFMSCs and to observe the biocharacteristics and the multi-potential of HAFMSCs after cryopreservation for the further fundamental researches and cl inical appl ications. Methods HAFMSCswere isolated from the amniotic fluid of pregnant women during the second trimester by the improved two-step method.HAFMSCs were cryopreserved with different cryopreservation protocols (containing different contents of FBS and DMSO atcryoprotectant) in l iquid nitrogen for 12 weeks. The biocharacteristics of the HAFMSCs after cryopreservation were analyzed. The growth characteristics were observed by MTT method and the growth curves were drawn. The surface antigens of HAFMSCs were detected using flow cytometry, including CD29, CD34, CD44, CD45, CD73, and CD90. The adi pogenic and osteogenic differentiation abil ities of HAFMSCs were observed. The mRNA levels of Oct-4 and Nanog of the HAFMSCs were compared between before and after cryopreservations. Results At 12 weeks after cryopreservation, different protocols had different effects on the cell viabil ity; the better formula of cryoprotectant was 50% DMEM, 40% FBS, and 10% DMSO. After cryopreservation, the cells proliferated rapidly and the growth curves showed “S” shape, which was the same as the cells before cryopreservation. Phenotype showed that HAFMSCs were positive for the surface markers CD29, CD44, CD73, and CD90, and negative for CD34 and CD45. After 21 days of adi pogenic differentiation, the l ipid droplets were observed by oil red O staining. After 21 days of osteogenic differentiation, the calcium mineralizations were verified by von Kossa staining. There was no significant difference (P gt; 0.05) in the mRNA levels of Oct-4 and Nanog between before and after cryopreservations. Conclusion HAFMSCs have rapid proliferation and multi-potential in vitro. The cells have high viabil ities and no changes of the biocharacteristics and differentiation potential ities after cryopreservation for 12 weeks. Cryoprotectant containing 50% DMEM, 40% FBS, and 10% DMSO is a better cryopreservation protocol.
ObjectiveTo improve the knowledge of primary tracheobronchial lymphoma.MethodsTwo patients with primary tracheobronchial lymphoma admitted to First Affiliated Hospital Guangxi Medical University in 2013 and in 2016 were analyzed retrospectively, and related literatures were reviewed. Chinese National Knowledge Infrastructure, Wanfang database and VIP database were searched by using " trachea lymphoma” or " bronchus lymphoma” as keywords. Meanwhile, databases including PubMed, Ovid Medline and Embase database were retrieved with " Trachea” or " Bronchus” AND " Lymphoma” as keywords. Seventy-two cases of primary tracheobronchial lymphoma were reported, the clinical feature, imaging feature, pathological feature, treatment and prognosis of 72 cases were analyzed.ResultsThe two patients were both females. The chief complaints included cough and dyspnea. The tissue biopsy under bronchoscope was the main diagnostic method. On histopathology, one case was diagnosed grade B cell lymphoma, another was diagnosed mantle cell lymphomas. After chemotherapy, the symptoms were relieved and never relapsed in the follow-up period until October 2016. Literature review found 72 patients with primary tracheobronchial lymphoma with women being the majority (47 cases, 65.28%). The patients aged from16 to 82 years with a median age of 51 years, and nine cases (12.5%) of the patients aged from 16 to 30 years. The specific clinical symptoms were cough and dyspnea. Pulmonary function prompted obstructive dysfunction of pulmonary ventilation. The chest CT scan showed consolidation shadow in trachea and bronchus with or without pulmonary atelectasis. Neoplasms could be found by bronchoscopy. Mucosa-associated lymphoid tissue (MALT) lymphoma was the most common pathological characteristic (36.11%). Mantle cell lymphomas has not been reported currently.ConclusionsPrimary tracheobronchial lymphoma is one of the rare airway tumors. Its clinical features, imaging examinations and bronchoscopic characteristics are not specific, which can be easily confused with lung cancer. It also can cause life-threatening airway obstruction. The bronchoscopy play an important role in diagnosis of primary tracheobronchial lymphoma, while the diagnosis is confirmed by histopathological examination. The main pathology type is MALT lymphoma. The prognosis is good.
The central aim of this experiment was to compare the articulatory and acoustic characteristics of students with normal hearing (NH) and school aged children with hearing loss (HL), and to explore the articulatory-acoustic relations during the nasal finals. Fourteen HL and 10 control group were enrolled in this study, and the data of 4 HL students were removed because of their high pronunciation error rate. Data were collected using an electromagnetic articulography. The acoustic data and kinematics data of nasal finals were extracted by the phonetics and data processing software, and all data were analyzed by t test and correlation analysis. The paper shows that, the difference was statistically significant (P<0.05 orP<0.01) in different vowels under the first two formant frequencies (F1, F2), the tongue position and the articulatory-acoustic relations between HL and NH group. The HL group’s vertical movement data-F1 relations in /en/ and /eng/ are same as NH group. The conclusion of this study about participants with HL can provide support for speech healing training at increasing pronunciation accuracy in HL participants.
In this paper, we propose a multi-scale mel domain feature map extraction algorithm to solve the problem that the speech recognition rate of dysarthria is difficult to improve. We used the empirical mode decomposition method to decompose speech signals and extracted Fbank features and their first-order differences for each of the three effective components to construct a new feature map, which could capture details in the frequency domain. Secondly, due to the problems of effective feature loss and high computational complexity in the training process of single channel neural network, we proposed a speech recognition network model in this paper. Finally, training and decoding were performed on the public UA-Speech dataset. The experimental results showed that the accuracy of the speech recognition model of this method reached 92.77%. Therefore, the algorithm proposed in this paper can effectively improve the speech recognition rate of dysarthria.
Objective To investigate the short-term effectiveness of transverse antecubital incision in the treatment of failed closed reduction of Gartland type Ⅲ supracondylar humeral fractures (SHFs) in children. Methods Between July 2020 and April 2022, 20 children with Gartland type Ⅲ SHFs who failed in closed reduction were treated with internal and external condylar crossing Kirschner wire fixation through transverse antecubital incision. There were 9 boys and 11 girls with an average age of 3.1 years (range, 1.1-6.0 years). The causes of injuries were fall in 12 cases and fall from height in 8 cases. The time from admission to operation ranged from 7 to 18 hours, with an average of 12.4 hours. The healing of the incision and the occurrence of complications such as nerve injury and cubitus varus were observed after operation; the elbow flexion and extension range of motion after removing the gypsum, after removing the Kirschner wire, and at last follow-up were recorded and compared, as well as the elbow flexion and extension and forearm rotation range of motion at last follow-up between healthy and affected sides; the Baumann angle was measured on the X-ray film, and the fracture healing was observed. At last follow-up, the effectiveness was evaluated according to the Flynn elbow function evaluation criteria. ResultsAll incisions healed by first intention, and there was no skin necrosis, scar contracture, ulnar nerve injury, and cubitus varus. Postoperative pain occurred in the radial-dorsal thumb in 2 cases. The gypsum was removed and elbow flexion and extension exercises were started at 2-4 weeks (mean, 2.7 weeks) after operation, and the Kirschner wire was removed at 4-5 weeks (mean, 4.3 weeks). All the 20 patients were followed up 6-16 months, with an average of 12.4 months. The fracture healing time was 4-5 weeks, with an average of 4.5 weeks, and there was no complication such as delayed healing and myositis ossificans. The flexion and extension range of motion of the elbow joint gradually improved after operation, and there were significant differences between the time after removing the gypsum, after removing the Kirschner wire, and at last follow-up (P<0.017). There was no significant difference in the flexion and extension of the elbow joint and the forearm rotation range of motion between the healthy and affected sides at last follow-up (P>0.05). There was no significant difference in Baumann angle between the time of immediate after operation, after removing the Kirschner wire, and at last follow-up (P>0.05). According to Flynn elbow function evaluation standard, 16 cases were excellent and 4 cases were good, the excellent and good rate was 100%. Conclusion The treatment of Gartland type Ⅲ SHFs in children with failed closed reduction by internal and external condylar crossing Kirschner wire fixation through transverse antecubital incision has the advantages of complete soft tissue hinge behind the fracture for easy reduction and wire fixation, small incision, less complications, fast fracture healing, early functional recovery, reliable reduction and fixation, and can obtain satisfactory results.