Identification of potential candidate proteins for reprogramming spinal cord-derived astrocytes into neurons: a proteomic analysis.

Our previous study has confirmed that astrocytes overexpressing neurogenic differentiation factor 1 (NEUROD1) in the spinal cord can be reprogrammed into neurons under in vivo conditions. However, whether they can also be reprogrammed into neurons under in vitro conditions remains unclear, and the mechanisms of programmed conversion from astrocytes to neurons have not yet been clarified. In the present study, we prepared reactive astrocytes from newborn rat spinal cord astrocytes using the scratch method and infected them with lentivirus carrying NEUROD1. The results showed that NEUROD1 overexpression reprogrammed the cultured reactive astrocytes into neurons in vitro with an efficiency of 13.4%. Using proteomic and bioinformatic analyses, 1952 proteins were identified, of which 92 were differentially expressed. Among these proteins, 11 were identified as candidate proteins in the process of reprogramming based on their biological functions and fold-changes in the bioinformatic analysis. Furthermore, western blot assay revealed that casein kinase II subunit alpha (CSNK2A2) and pinin (PNN) expression in NEUROD1-overexpressing reactive astrocytes was significantly increased, suggesting that NEUROD1 can directly reprogram spinal cord-derived reactive astrocytes into neurons in vitro, and that the NEUROD1-CSNK2A2-PNN pathway is involved in this process. This study was approved by the Animal Ethics Committee of Fujian Medical University, China (approval No. 2016-05) on April 18, 2016.

Social support and the self-rated health of older people: A comparative study in Tainan Taiwan and Fuzhou Fujian province.

The lack of social support in elderly populations incurs real societal costs and can lead to their poor health. The aim of this study is to investigate the self-rated health (SRH) and social support among older people as well as its associated factors.We conducted a cross-sectional study among 312 urban community-dwelling elderly aged 65 to 90 years in Tainan Taiwan and Fuzhou Fujian Province from March 2012 to October 2012. A Spearson correlation test, independent t test, a Pearson χ test, a linear regression analysis, and a multiple-level model were performed to analyze the results.The participants identified children as the most important source of objective and subjective support, followed by spouse and relatives. Tainan's elderly received more daily life assistance and emotional support, showed stronger awareness of the need to seek help, and maintained a higher frequency of social interactions compared with the elderly in Fuzhou. The mean objective support, subjective support, and support utilization scores as well as the overall social support among Tainan's elderly were significantly high compared with the scores among Fuzhou's elderly. Further, Tainan's elderly rated better SRH than Fuzhou's elderly. Correlation analysis showed that social support was significantly correlated with city, age, living conditions, marital status, and SRH. Multiple linear regression analysis, with social support as a dependent variable, retained the following independent predictors in the final regression model: city (4.792, 95% confidence interval [CI]: 3.068-6.516, P = 0.000), age (-0.805, 95% CI: -1.394 to -0.135, P = 0.013), marital status (-1.260, 95% CI: -1.891 to -0.629, P = 0.000), living conditions (4.069, 95% CI: 3.022-5.116, P = 0.000), and SRH -1.941, 95% CI: -3.194 to -0.688, P = 0.003). The multiple-level model showed that city would impact older people's social support (χ = 5.103, P < 0.001). Marital status (-2.133, 95% CI: -2.768 to -1.499, P = 0.000), education (1.697, 95% CI: 0.589-2.805 P = 0.003), living conditions (4.20, 95% CI: 1.762-6.638, P = 0.000), and SRH (-3.144, 95% CI: -4.502 to -1.727, P = 0.000) were the associated factors. Thus, city, age, marital status, education, living conditions, and SRH might be the associated factors for social support among older people.This study presents some feasible implications for social support improvement in China and in other nations worldwide.

[Effect of C5a on expression of thrombomodulin in endothelial cells in vitro].

OBJECTIVE: To investigate the effect of C5a on the expression of thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs cultured in vitro were stimulated with C5a for 8, 12, 16, 20 hours in a concentration of 200 microg/L, and also with different concentrations of 100, 200, 300 microg/L for 12 hours respectively. The levels of both mRNA and protein expression of TM was detected by real-time polymerase chain reaction (PCR) and Western blotting respectively, and the dose and time dependent effects of C5a on the expression of TM were evaluated. RESULTS: C5a down-regulated the TM expression at both protein and mRNA level. The down-regulation was time-dependent [(93.11+/- 1.57) x10(-2) , (71.05+/-3.39)x10(-2), (65.48+/-4.28)x10(-2), (62.69+/-4.03)x10(-2)at protein level and (301.71+/-80.40)x10(-6), (38.29+/-20.24)x10(-6), (8.82+/-2.66)x10(-6), (7.05+/-0.80)x10(-6) at mRNA level, all P<0.05] and dose-dependent [(113.25+/-3.97)x10(-2), (80.18+/-2.56)x10(-2), (73.22+/- 4.36)x10(-2) at protein level and (401.77+/-20.46)x10(-6), (31.12+/-3.51)x10(-6), (18.19+/-1.46)x10(-6) at mRNA level , all P<0.05]. When concentration of C5a at 300 microg/L was used to stimulate HUVECs for longer than 12 hours, the lowering of TM at protein level was slowed down obviously. And concentration of C5a at 200 microg/L was used to stimulate HUVECs for 12 hours, the lowering of TM at mRNA level was slowed down obviously. CONCLUSION: C5a can depress the gene expression of TM, and then affect the protein's translation. By this means, C5a can lead to hypercoagulability and inflammatory injuries in sepsis.

[Forty-three tracheal stricture with Ni-Ti shape memory alloy stent].

OBJECTIVE: To evaluate the application of Ni-Ti shape memory alloy stent for treating tracheal stricture, including its indications and complications. METHODS: Forty-three patients clinical data were analyzed, among them, 15 suffered from tumor; 7 trauma; 4 granulation; 1 Wegener's granuloma, 2 rhinoscleroma, 1 without clear diagnosis; 7 were relapsing polychondritis; 1 were respiratory amyloidosis; 4 were tracheobronchial tuberculosis, 5 were tracheobronchomalacia. The stricture degree: Level I: 15 patients, Level II:24 patients, Level III:4 patients. The location of stricture in 5 patients extended to the subglottis area. All patients had tracheal stricture and accepted the treatment with Ni-Ti shape memory alloy stent. RESULTS: Forty-two patients breathed freely after the operation, 1 patient died after surgery. The tracheostoma in 18 patients was closed 34 days after operation. Among the remaining 8 patients with tracheostoma:3 patients left hospital with tracheal cannula; 1 patient died,2 patients were found stent shifted and no evident breath improving; 2 patients were found granulation and had secondary operation with laser. CONCLUSION: Ni-Ti shape memory alloy stent can expand the tracheal stricture quickly and thus improve the breath effectively, which is minimally invasive and simple. It can be used as a method in the treatment of tracheal stricture.

[Complications of tracheoesophageal puncture for Blom-Singer voice restoration after total laryngectomy].

OBJECTIVE: To investigate causes and treatment of complications of tracheoesophageal puncture for Blom-Singer voice restoration after total laryngectomy. METHODS: From 1986 to 2004, one hundred and fifty one cases with Blom-Singer technique after total laryngectomy for voice restoration were retrospectively analyzed. RESULTS: All cases were followed up from 6 months to 15 years. Among 151 cases, 138 cases got successful phonation and the total success rate of voice restoration was 91.4%. Of the 151 cases, 15 cases occurred complications. The rate of complications was 9.9%. The common complications included fistula granulations, infection, and leakage. Twelve cases got successful treatment accordingly, and the other 3 cases failed in phonation. CONCLUSIONS: The procedure of tracheoesophageal puncture for voice restoration is relatively simple and has low complications. this method had high phonation success and good phonation quality, which is one of the best way to make laryngectomee to speak.

[Pharyagoesophageal sphinoter myotomy for voice rehabilitation after total laryngectomy].

OBJECTIVE: In those patients received total laryngectomy, Blom-Singer prosthesis speech appears to be the more effective form of rehabilitation. To reduce the effect of pharyngoesophageal sphincter on the Blom-Singer prosthesis voice restoration following total laryngectomy, the pharyagoesphageal sphincter myotomy was designed and carried out. METHODS: After total laryngectomy, the cricopharyngeus muscle stricture or spasm appears to inhibit the air flowing and prevent the speech. The pharyngoesophageal sphincter is now defined as the area including the cricopharyngeus with a portion of inferior constrictor, as well as some of the upper cervical esophagus. The pharyngoesophageal sphincter were incised intra- and post- operation of total laryngectomy, the dissection was approximately 5 cm in vertical length and 1 cm in width. RESULTS: Among the 33 patients who received pharyngoesophageal sphincter myotomy following total laryngectomy, voice restoration was achieved in 32 cases, the success rate was 97%. 25 patients had survived for 3 years, 18 patients survived for 5 years. CONCLUSION: Pharyagoesophageal sphincter myotomy can raise the success rate of rehabilitation of the voice by Blom-Singer prosthesis.

[Surgical management of metastatic disease in the conjunctive area between neck and thorax].

OBJECTIVE: To study the surgical management of metastatic disease in the conjunctive area between the neck and thorax and its efficacy. METHODS: Fourteen cases with metastatic node disease in the area between neck and thorax were collected and analysed. Eleven tumors were from the thyroid cancer, and the other three were from the hypopharyngeal cancer, esophagual cancer and malignant pheochromocytoma, respectively. The clavicle was displaced or resected, and the upper half of the manubrium might also be resected when necessary. The recurrent laryngeal nerve and phrenic nerve were exposed and protected. The metastatic disease was completely removed with the internal jugular and/or the brachiocephalic vein resected or spared, depending on the disease condition. RESULTS: In 10 cases with metastases from the thyroid, no local recurrence was found within the follow-up period from 2 to 5 years. In contrast, no patient with metastatic disease from hypopharyngeal or esophageal cancer survived more than 11 months. No serious complications were found in this group. CONCLUSIONS: The surgical treatment of node metastases in the conjunctive area between neck and thorax from the well-developed thyroid cancer has promising effect and is comparatively safe.

[Culture and identification of dendritic cells from the peripheral blood of patients with laryngeal squamous cell carcinoma in vitro].

OBJECTIVE: To culture dendritic cells (DC) from peripheral blood of patients with laryngeal carcinoma for therapeutic aid. METHODS: Adherent peripheral blood mononuclear cells from peripheral blood were cultured with 15 ng/ml rhGM-CSF and 7 ng/ml rhIL-4 for one or two weeks. The purity of DC was detected by immunocytochemistry method. The mixed leukocyte reactions stimulated by DC loaded with laryngeal carcinoma antigen were tested by measuring 3H-TdR uptake. RESULTS: A considerable number of suspended cells with spicular or dendritic appearance were observed after 1 week of culture, and their mitochondria were rich in cytoplasm. The positivity of DC was about 30%-60%. DC loaded with laryngeal antigen could induce proliferation of syngeneic T lymphocytes. CONCLUSION: A large number of DC with high purity can be cultured from peripheral blood of patients with laryngeal carcinoma in vitro. It may be used in further experimental studies for clinical applications.

[Analysis of clinical manifestations of pharyngeal and laryngeal amyloidosis].

OBJECTIVE: To study the clinical manifestations of pharyngeal and laryngeal amyloidosis. METHODS: Retrospective analysis of the clinical data of the inpatients with systemic and local amyloidosis. RESULTS: Of the 25 cases, sex ratio was 21:4, and average age was 48 years. Seventeen cases were systemic, of which 7 patients died in 2 years since diagnoses were made. One case was local bronchial amyloidosis. Seven cases were local primary pharyngeal and laryngeal amyloidosis and no affirmative death report was recorded since diagnoses were made. CONCLUSIONS: Amyloidosis occurs more likely in male and middle- and old-aged patients. Pharyngeal and laryngeal local primary amyloidosis have relatively better prognosis than that of systemic amyloidosis. Follow-up is important because local recurrence may be happened.