Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective:To explore the effect of behavioral training of creative art therapy(CAT) on interpersonal communication in teenagers.Methods:Eighty teenagers were randomly divided into study group( n=40) and control group( n=40).The study group was received the behavioral training of CAT (12 times, each activity lasts for 120 min), and the control group did not take part in the behavioral training of CAT.The two groups were measured with self-esteem scale(SES), shyness scale(SS) and social anxiety scale for children(SASC) before and after behavior training.SPSS 17.0 software was used for statistical analysis, and t-test variance analysis and Mann-Whitney U test were used to compare the differences between groups or within groups. Results:Before and after behavioral training, changes in the SES total score (7.00(5.00, 9.75) vs 1.00(0, 2.00)), the SS total score (7.00(6.00, 12.75) vs 2.00(1.00, 3.00)), the SASC total score (5.00(2.00, 8.75) vs 0(0, 1.00)) and two factor scores (3.00(0.25, 5.00) vs 1.00(0, 1.00), 2.00(1.00, 4.00) vs 0(0, 1.00))in the study group were obviously higher than those in the control group( Z=-7.347, -7.338, -5.841, -4.122 and -5.393, all P<0.05).The score differences of the SASC total score ((7.19±3.82) vs (3.21±3.04)) and two factor scores ((4.00±2.23) vs (1.68±2.05), 3.00 (1.00, 5.00) vs 2.00 (0, 2.00)) of girls were significantly higher than those of boys in the study group ( t=-3.611, -3.398, Z=-2.111, P=0.001, 0.002, 0.035). Conclusion:Behavioral training of CAT can improve teenagers' self-esteem, relieve their shyness and social anxiety, as well as promote their interpersonal communication.

Objective:To investigate the effect of art behavior intervention of yoga training on patients with insomnia comorbid depressive disorder.Methods:Totally 66 patients with insomnia comorbid depressive disorder were randomly divided into the study group( n=36) and the control group( n=30). All patients in two groups were given pharmacotherapy, while the patients in the study group were received art behavior intervention of yoga training for 20 times.Using self-made general information questionnaire to collect the patients' demographic and clinical characteristics.Polysomnography(PSG) was performed to measure objective sleep parameters.The Pittsburgh sleep quality index(PSQI), Hamilton depression scale(HAMD) and Hamilton anxiety scale(HAMA) were used to measure subjective sleep quality, depressive symptoms and anxiety symptoms at baseline and at the end of the interventions. Results:The score differences of pre-and post-intervention in the two group of PSQI total score((11.00±2.72) vs(9.00±2.22)), habitual sleep efficiency((2.02±0.81) vs (1.53±0.77)), daytime dysfunction((1.75±0.90) vs(0.96±0.85)), HAMD total score((12.19±6.25) vs(8.03±5.67)) and HAMA total score(13.00(8.25, 16.75) vs 7.00(5.00, 9.25))in the study group were obviously higher than those in the control group( t=3.220, 2.516, 3.595, 2.806, Z=-3.616, all P<0.05). Conclusion:Art behavior intervention of yoga training can improve sleep quality, depression and anxiety symptoms in patients with insomnia comorbid depressive disorder, which will be applied and popularized in clinical practice.

Objective: To investigate the effect of cough assist on sputum excretion and the outcome of withdrawal of mechanical ventilation after mechanically ventilated chronic obstructive pulmonary disease (COPD) patients with cough weakness. Methods: From January 2017 to December 2018, 74 patients with cough and weakness COPD after extubation of mechanical ventilation in the Department of Respiratory Critical Care Medicine of Hunan Provincial People's Hospital were divided into control group(n=37) and observation group (n=37) according to the random number table method. The patients in the control group were treated routinely after weaning and extubation, and the observation group was treated with cough assist after withdrawal of mechanical ventilation and extubation on the basis of the control group. The differences in drainage effect, blood gas index, reintubation rate and early prognosis index between the two groups were compared. Results: After the intervention treatment, the first active sputum excretion and the total sputum volume on the first day were (5.6±3.4) ml and (33.1±5.2) ml in the observationgroup, and (4.2 ±2.0) ml and (29.1±7.4) ml in the control group, the difference was statistically significant (t=-2.10, 2.875, P<0.05). The number of significant cases of respiratory sound improvement in the observation group and the control group was 21 and 14 cases, respectively, and the difference between the two groups was statistically significant (Z=-1.974, P < 0.05). The oxygen partial pressure (PaO₂) and oxygenation index (PaO₂/FiO₂) carbon dioxide partial pressure (PaCO₂) values of the observation group were (80.0±8.4), (345.9±19.2), (46.7±6.6)mmHg, and (74.8±9.1), (310.7±21.9), (50.9±7.1)mmHg in the control group. The difference was statistically significant (t=-2.504,-2.710, 2.579, all P<0.05). The reintubation rate, noninvasive ventilation time, and hospitalization days after the initial extubation in the observation group were as follows: 5.6%(2/36), (64.1±18.9)h, (6.0±1.7)d, and 22.2%(8/36), (76.7±15.3)h, (7.2±2.8)d in the control group. The difference was statistically significant (χ²=4.181, t=2.528, 2.438, all P<0.05). The non-invasive ventilation rate within 72h in the observation group and the control group were 63.9% (23/36) and 75.0% (27/36), the difference was not statistically significant (χ²=0.222, P>0.05). Conclusions The application of cough assist in RICU patients with cough weakness after extubation by mechanical ventilation can improve the expectoration efficiency of cough, improve oxygenation, reduce carbon dioxide retention, reduce the rate of re-intubation, shorten the time of noninvasive ventilation and hospitalization after extubation, and improve the curative effect.

Objective To investigate the cost-effectiveness of cefoperazone sulbactam sodium in treating nosocomial infection (lower respiratory tract infection).Methods 80 cases of nosocomial infection in Jincheng Second People's Hospital from January 2014 to January 2017 were treated as the subjects: the observation group was treated with cefoperazone sulbactam sodium and the control group was treated with cefodizime sodium.The data of two groups of patients were recorded and the data were analyzed statistically.The cost-effectiveness of cefoperazone sulbactam sodium in hospital infection was discussed.Results There was no significant difference in the clinical curative effect between the two groups.The cost of observation group (cefoperazone sulbactam sodium) was lower than that of the control group (cefodizime sodium), the difference was statistically significant (P< 0.05).Conclusion Patients with nosocomial infection choose to use cefoperazone sulbactam sodium as the treatment method, which has exact clinical efficacy, high cost-effectiveness.It is worthy of clinical wide application.

Objective To evaluate the role and mechanism of ifenprodil, which is the selective antagonist of N-methyl-D-aspartic acid subtype receptor NR2B, in soflurane-induced cognitive dysfunction in neonatal rats.Methods Twenty-eight 7-day-old Sprague Dawley rats, weighing 15-18 g, were randomly divided into 4 groups (n=7 each): control group (group C), ifenprodil group (group I), sevoflurane group (group S) and ifenprodil+sevoflurane group (group IS).Normal saline 0.2 ml was injected intraperitoneally in group C.Specific NR2B receptor antagonist ifenprodil 5 mg/kg was injected intraperitoneally at the corresponding time points in group I.Normal saline 0.2 ml was injected intraperitoneally and 2.0% sevoflurane was inhaled for 4 h in group S.Ifenprodil 5 mg/kg was injected intraperitoneally 2 h before sevoflurance inhalation, and 2.0% sevoflurance was inhaled for 4 h in group IS.The rats were then sacrificed 3 weeks after administration, their brains were immediately removed and hippocampal slices were prepared for electrophyisological experiments.The value of population spike amplitude (PSA) and long-term potentiation (LTP) were measured every 10 minutes.Induced LTP was recorded.Results Compared with group C, the values of PSA and rates of induced LTP were significantly decreased in group S (P＜0.01).The values of PSA and rates of induced LTP were significantly increased in group IS than those in group S (P＜0.01).Conclusion NR2B receptor is involved in sevoflurance-induced cognitive dysfunction in the neonatal rats.Pretreatment with ifenprodil 5.0 mg/kg can improve the neurotoxicity and protect the brain.

Objective To compare clinical efficacy of clobetasol propionate ointment and vitamin A acid cream in the treatment of skin papule type amyloidosis.Methods 100 cases of skin papule type amyloidosis were randomly divided into the observation group and control group,each had 50 patients.The control group was treated with Vitamin A acid cream while the observation group used clobetasol propionate ointment for treatment.Skin lesion area,infiltration,skin color,skin itching score as well as the cure rate and efficiency were compared after 4 weeks of treatment.Results The two groups' symptoms was improved,symptom scores was decreased gradually after treatment than before,the observation group's symptom scores at 1,2,3,4 weeks were (9.35 ± 1.88),(6.54 ±2.16),(4.08 ±1.32),(2.04 ± 0.95) points which was significant better than (10.86 ± 2.08),(7.98 ± 2.57),(6.25 ± 1.44),(4.56 ± 1.18) points of the control group,the difference of two groups was statistically significant (t =6.22,6.71,7.30,7.41,all P ＜ 0.05) ;4 weeks after treatment,the observation group's efficience and cure rates were 94％ and 34％,significantly higher than 70％ and 22％ of the control group,the difference was statistically significant (x2 =9.040,8.391,all P ＜ 0.05).Conclusion Clobetasol propionate ointment has a exact effect in treatment of skinpapule type amyloidosis than vitamin A acid cream,which is worthy of clinical application.

Neutrophils play a critical role in ventilation-induced lung injury. This study was aimed to investigate the characteristics of neutrophils influx in lungs induced by high tidal volume ventilation. Anaesthetized rats were randomly divided into low tidal volume ventilation group (Vt: 7 mL/kg, LV group) or high tidal volume ventilation group (Vt: 42 mL/kg, HV group) (n=40 in each). Rats in each group were ventilated for 0, 60, 90, 120 and 240 min. The wet/dry lung weight ratio (W/D) was measured. The levels of macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α), and the activity of myeloperoxidase (MPO) were detected by enzyme-linked immunosorbent assay (ELISA). The number of neutrophils in bronchoalveolar lavage fluid (BALF) was counted after Wright's staining, and the percentage of netrophils in lung tissues calculated. Histopatholgical examination was used to observe the changes of lung tissues after different ventilations. The results showed that the W/D weight ratio was increased, and the levels of MIP-2 and TNF-α significantly enhanced in HV group at 90, 120 and 240 min. Neutrophils in BALF and the neutrophil percentage in lung tissues were also elevated at 120 and 240 min, which coincided with the enhanced activity of MPO in HV group. The lung injury was significantly related with the ventilation time and the infiltration of neutrophils in lungs in HV group. In conclusion, in ventilation-induced lung injury, neutrophil infiltration is present in a time-dependent manner and associated with the aggravated lung injury. Pulmonary structural damage may be the main reason for ventilation-induced lung injury.
Animals ; Lung ; physiopathology ; Lung Injury ; physiopathology ; Male ; Neutrophil Infiltration ; physiology ; Neutrophils ; physiology ; Rats ; Rats, Sprague-Dawley

Neutrophils play a critical role in ventilation-induced lung injury. This study was aimed to investigate the characteristics of neutrophils influx in lungs induced by high tidal volume ventilation. Anaesthetized rats were randomly divided into low tidal volume ventilation group (Vt: 7 mL/kg, LV group) or high tidal volume ventilation group (Vt: 42 mL/kg, HV group) (n=40 in each). Rats in each group were ventilated for 0, 60, 90, 120 and 240 min. The wet/dry lung weight ratio (W/D) was measured. The levels of macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α), and the activity of myeloperoxidase (MPO) were detected by enzyme-linked immunosorbent assay (ELISA). The number of neutrophils in bronchoalveolar lavage fluid (BALF) was counted after Wright's staining, and the percentage of netrophils in lung tissues calculated. Histopatholgical examination was used to observe the changes of lung tissues after different ventilations. The results showed that the W/D weight ratio was increased, and the levels of MIP-2 and TNF-α significantly enhanced in HV group at 90, 120 and 240 min. Neutrophils in BALF and the neutrophil percentage in lung tissues were also elevated at 120 and 240 min, which coincided with the enhanced activity of MPO in HV group. The lung injury was significantly related with the ventilation time and the infiltration of neutrophils in lungs in HV group. In conclusion, in ventilation-induced lung injury, neutrophil infiltration is present in a time-dependent manner and associated with the aggravated lung injury. Pulmonary structural damage may be the main reason for ventilation-induced lung injury.