Objective:To explore the genetic diversity of Mycobacterium tuberculosis (MTB) in different types of tuberculosis and its association with clinical features, providing evidence for precise diagnosis and treatment of tuberculosis. Methods:This cross-sectional study included 103 cases of tuberculosis (38 with simple pulmonary tuberculosis, 43 with tuberculous pleurisy, and 22 with pulmonary combined with extrapulmonary tuberculosis) from Shenzhen Third People′s Hospital from 2015 to 2018. Paired bacterial strains from lung and pleural effusion/extrapulmonary sites were collected. Whole-genome sequencing (WGS) was used for drug resistance prediction, and genetic diversity (π value) was calculated as well as differential genes screening. Statistical analysis included paired t-tests and χ2 tests to compare clinical, bacteriological and genetic diversity features among groups.Results:The simple pulmonary tuberculosis group exhibited significantly higher rates of retreatment (71.7%, 27/38), cavitation (70.4%, 19/27), and multidrug-resistant or rifampicin-resistant (MDR/RR) (60.5%, 23/38) compared to the tuberculous pleurisy group (retreatment 11.9%, 5/42; cavitation 11.9%, 5/42; MDR/RR 16.3%, 7/43) and extrapulmonary tuberculosis group (retreatment 9.1%, 2/22; cavitation 18.2%, 4/22; MDR/RR 13.6%, 3/22) ( P<0.05). The overall π values of the MTB strain genomes in lung [(5.94±3.93)×10 ?5], pleural effusion[(6.22±3.51)×10 ?5], and extrapulmonary tissues [(5.83±3.54)×10 ?5] showed no significant differences ( H=0.10, P=0.94). Differential gene diversity analysis revealed that π value alternating genes related to respiration and intermediate metabolism were prominently high [tuberculous pleurisy 32.4% (11/34) and extrapulmonary tuberculosis groups 31.4% (32/102)], while cell wall-associated genes dominated in the simple pulmonary tuberculosis group (42.9%, 6/14). Drug resistance profiles and mutation spectra were identical across isolates from different sites within the same patient. Conclusion:WGS revealed the MTB diversity among different types of tuberculosis. Difference between pulmonary and extrapulmonary environments may impel the adaptive alternations of the bacterial strains to maintain survival with higher overall genome stability. Drug resistance testing of lung-derived isolates may provide references on extrapulmonary tuberculosis treatment.

Objective:To explore the genetic diversity of Mycobacterium tuberculosis (MTB) in different types of tuberculosis and its association with clinical features, providing evidence for precise diagnosis and treatment of tuberculosis. Methods:This cross-sectional study included 103 cases of tuberculosis (38 with simple pulmonary tuberculosis, 43 with tuberculous pleurisy, and 22 with pulmonary combined with extrapulmonary tuberculosis) from Shenzhen Third People′s Hospital from 2015 to 2018. Paired bacterial strains from lung and pleural effusion/extrapulmonary sites were collected. Whole-genome sequencing (WGS) was used for drug resistance prediction, and genetic diversity (π value) was calculated as well as differential genes screening. Statistical analysis included paired t-tests and χ2 tests to compare clinical, bacteriological and genetic diversity features among groups.Results:The simple pulmonary tuberculosis group exhibited significantly higher rates of retreatment (71.7%, 27/38), cavitation (70.4%, 19/27), and multidrug-resistant or rifampicin-resistant (MDR/RR) (60.5%, 23/38) compared to the tuberculous pleurisy group (retreatment 11.9%, 5/42; cavitation 11.9%, 5/42; MDR/RR 16.3%, 7/43) and extrapulmonary tuberculosis group (retreatment 9.1%, 2/22; cavitation 18.2%, 4/22; MDR/RR 13.6%, 3/22) ( P<0.05). The overall π values of the MTB strain genomes in lung [(5.94±3.93)×10 ?5], pleural effusion[(6.22±3.51)×10 ?5], and extrapulmonary tissues [(5.83±3.54)×10 ?5] showed no significant differences ( H=0.10, P=0.94). Differential gene diversity analysis revealed that π value alternating genes related to respiration and intermediate metabolism were prominently high [tuberculous pleurisy 32.4% (11/34) and extrapulmonary tuberculosis groups 31.4% (32/102)], while cell wall-associated genes dominated in the simple pulmonary tuberculosis group (42.9%, 6/14). Drug resistance profiles and mutation spectra were identical across isolates from different sites within the same patient. Conclusion:WGS revealed the MTB diversity among different types of tuberculosis. Difference between pulmonary and extrapulmonary environments may impel the adaptive alternations of the bacterial strains to maintain survival with higher overall genome stability. Drug resistance testing of lung-derived isolates may provide references on extrapulmonary tuberculosis treatment.

Esophageal Achalasia/surgery* ; Esophagoscopy ; Humans ; Muscles ; Myotomy ; Tissue Adhesions

Objective:To evaluate the long-term efficacy and safety of a novel self-help inflatable balloon to prevent esophageal stenosis after extensive endoscopic submucosal dissection (ESD).Methods:Patients with early esophageal cancer or precancerous lesions, undergoing ESD in the First Medical Center of Chinese PLA General Hospital from January 2018 to December 2019 were included in the prospective study, who had post-ESD mucosal defect greater than 5/6 of the esophageal circumference and 30-100 mm in length. The self-help inflatable balloon was used to prevent esophageal stenosis after ESD. Mucosal defect of ESD was divided into grade 1 (≥5/6 and less than the whole circumference) and grade 2 (the whole circumference). The incidence of stricture, the time from ESD to the occurrence of stricture, the total number of endoscopic balloon dilations (EBD) or radial incision and cuttings (RIC), and other adverse events were observed.Results:A total of 27 patients met the including criteria with follow-up time of 14-38 months, including 3 patients of grade 1 and 24 of grade 2. The ulcer longitudinal length was 73.7±18.4 mm. The time of wearing balloons was 92.0±20.0 days. The overall frequency of stricture was 18.5% (5/27), and the stricture incidence of patients of grade 2 resection was only 16.7% (4/27). The median time from balloon removal to stricture was 17 days. To treat the stricture, two patients received 3 EBD sessions, and three other patients received 2, 1 and 2 RIC sessions, respectively. No balloon was removed in advance, and none had a perforation or delayed bleeding.Conclusion:The self-help inflatable balloon shows high efficacy and safety in preventing esophageal stenosis in patients with mucosal defect greater than 5/6 of the esophageal circumference and less than 100 mm in length after extensive esophageal ESD.

Objective:To investigate the expression and cellular provenance of interleukin 17A (IL-17A) in non-eosinophilic chronic rhinosinusitis with nasal polyps (nECRSwNP), and to analyze the possible reasons for its different expression.Methods:Samples were collected from 14 patients with eosinophilic chronic rhinosinusitis with nasal polyps (ECRSwNP) and 28 patients with nECRSwNP, who underwent functional endoscopic sinus surgery in the Third Affiliated Hospital of Sun Yat-sen University from January 2017 to May 2018, including 33 males and 9 females, with the age ranging from 18 to 65 years old. Enzyme linked immune sorbent assay (ELISA) and flow cytometry were used to investigate the expression and cellular origins of IL-17A in the nasal tissue of ECRSwNP and nECRSwNP groups. Then the difference of quantity and differentiation ability of the major cells producing IL-17A between ECRSwNP and nECRSwNP groups were analyzed by flow cytometry. Finally, the expressions of IL-6, transforming growth factor-β(TGF-β), and IL-23, which were considered as the important factors in promoting Th17/Tc17 differentiation in CRSwNP and their correlation with IL-17A, were analyzed by ELISA. Statistical analysis was performed using IBM SPSS 20.Results:The IL-17A protein levels and IL-17A +lymphocyte percentages were higher in nECRSwNP group compared with that of the ECRSwNP group (158.56 (167.76) pg/ml ( M( QR)) vs. 9.42 (11.33) pg/ml, 10.21%±1.54% ( ± s) vs. 3.93%±0.80%, Z=2.95, t=3.62, all P<0.01). Tc17 cells (CD8 +T cells producing IL-17A) and Th17 cells (CD4 +T cells producing IL-17A) were major IL-17A producers in both ECRSwNP and nECRSwNP group. Further analysis revealed that there was no significant difference in quantity of CD8 +and CD4 +T cells between ECRSwNP and nECRSwNP group, but the differentiation ability about CD8 +and CD4 +T cells differentiating into Tc17 and Th17 in nECRSwNP group was stronger than that in ECRSwNP. The high expressions of IL-6 and TGF-β, which were considered as the important factors in promoting Th17/Tc17 differentiation were also found in nECRSwNP group compared with ECRSwNP (56.07 (234.25) pg/ml vs. 8.27 (12.51) pg/ml, (5.44±0.34) pg/ml vs. (4.17±0.22) pg/ml, Z=2.426, t=2.29, all P<0.05). But the difference in expression of IL-23 was not significant difference between the two groups. Moreover, the expression of IL-17A showed significantly positive correlation with IL-6 ( r=0.615, P=0.009). No positive correlation between IL-17A and TGF-β or IL-23 was observed. Conclusions:The expression of IL-17A in nasal mucosa of nECRSwNP patients is significantly higher than that of ECRSwNP, which is due to the increase of expression and differentiation of Tc17/Th17 cells. IL-17A shows positive correlation with IL-6 in CRSwNP, which is the important factor in promoting Th17/Tc17 differentiation.

Objective To study the effect of age-related white matter changes (ARWMC) on first symptomatic ischemic stroke events in the oldsters. Methods For the prospective study, a total of 368 eligible oldsters were enrolled in the study from January 2010 to August 2012. The degrees of ARWMC were assessed by ARWMC scale;according to the scores, they were divided into non ARWMC group, mild-moderate ARWMC group and severe ARWMC group. The patients were followed up once every 3 months. The clinical endpoint events and time (first symptomatic ischemic stroke, myocardial infarction and all-cause death) were recorded. Analyses of variance and Chi-square test were used to compare the differences of clinical data among the 3 groups. COX regression was used to assess the risk differences of first symptomatic ischemic stroke in the oldsters of three groups. Results After an average of follow-up for 48.7 months, 50 participants (13.6％) had first symptomatic ischemic stroke;25 (25.8％) were categorized as the severe ARWMC group, 22 (10.9％) were as the mild-medium group, and 3 (4.4％) were as the non ARWMC group. Among the three groups, the differences in age, history of hypertension, systolic blood pressure, incidence of clinical endpoint events and first symptomatic ischemic stroke, and follow-up time of endpoint events were statistically significant (P<0.05); patients from the severe ARWMC group were the oldest, and had the longest history of hypertension, the highest systolic blood pressure, the highest incidence of clinical end events and first symptomatic ischemic stroke, and the shortest follow-up period for clinical end events. COX regression analysis showed that the risk of first symptomatic ischemic stroke in the severe ARWMC group was about 8 times higher than that in the non ARWMC group (hazard ratio=9.012, 95％CI: 2.310-35.154, P=0.002). Conclusion In oldsters, severe ARWMC often accompany hypertension history and poor blood pressure controll, and it is an independent and serious risk factor for long-term first symptomatic ischemic stroke.

Objective To investigate the nursing of sedation and analgesia using low-dose fentanyl with midazolam during intra-arterial treatment with mechanical thrombectomy by means of stent for patients with anterior-circulation acute ischemic stroke.Methods All of 61 patients who underwent intra-arterial treatment with anterior-circulation acute ischemic stroke were retrospectively collected.They were divided into two groups according to the different program of individual conscious sedation:the control group(30 cases)used midazolam for intravenous maintain sedation,experimental group(31 cases)added the low-dose fentanyl on the basis of the control group.The thrombectomy was performed by the same group of interventional radiologists,while the effect of sedative,vital signs,respiratory curve and SpO2(blood oxygen saturation),assessment of Ramsay's sedation score,restlessness and other sedation adverse reactions were respectively observed in both groups.Nursing intervention was performed aiming at different states of patients during operation.Results All 61 patients were implemented with good medical care with no intracranial hemorrhage or intraoperative death.In the control group,there were 4 patients with Ramsay 1 point of sedation,resulted motion artifacts in digital subtraction angiography images in 3 of them,1 patient with slight reduction of SpO2,and vomiting in 4 patients.1 patient with decreased mean arterial pressure.In the experimental group,there was no Ramsay 1 point of sedative patient.The SpO2 was decreased in 5 patients.Respiratory inhibition occurred in 3 patients and the mean arterial pressure decreased slightly in the other 3 patients.Conclusions Application of low-dose fentanyl and midazolam is safe and reliable in patients who underwent intra-arterial treatment with anterior-circulation acute ischemic stroke.While closely observation to the changes of breath frequency curve and SpO2 in operation are very essential to actively prevent the occurrence of respiratory depression.

Objective:The aim of the current study is to understand the sleep disorders in children with autism and related factors.Methods:A case-control study method was conducted in this study which included 198 children autism who met the criteria of the Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-Ⅳ) and 233 cases of gender-and age-matched normal children.Their general demographic and sleep data were collected and the children with autism were assessed with Autism Behavior Checklist,Child Behavior Checklist and Autism Spectrum Screening Questionnaire.Correlation analysis and regression analysis were performed to compare sleep quality,sleep problems and the related factors of the two groups.Results:The scores of difficulty falling asleep,sleep restlessness,taking hold or shaking,restless sleep,interrupted sleep,nightmares,sleepwalking,night terrors,sleep snoring,nocturnal enuresis,sleep apnea,mouth breathing,sweating and sleep scores were higher in the autism group compared to the normal control group before falling asleep (P ＜ 0.05),while the proportion of daytime sleep time in the autism group was lower than the normal control group (P ＜0.05).With sleep status as dependent variable,linear regression analysis was conducted with children's birth weight,walking month age,speaking month age,father age,and autism spectrum as independent variables.The multiple linear regression analysis showed the scores of,sensory behaviors,body and object use behaviors,and self-help behaviors in Autism Behavior Checklist for autistic children were positively associated with their sleep (β =0.21,0.21,0.29,P ＜ 0.05).However,paternal age at birth in the autism group had negative associatin with the sleep of autistic children (β =-0.16,P ＜0.05).In the autism group,their total scores of Autism Behavior Checklist and the scores of language and social behavior in Autism Behavior Checklist were not associated with their sleep status score (P ＞ 0.05).Conclusion:It suggests that children with autism may have more sleep problems,and their sleep problems are related to autism symptoms including the scores of,sensory behaviors,body and object use behaviors,and self-help behaviors in Autism Behavior Checklist for autistic and their paternal age at birth.

Objective]In order to study the impact of adenoid hypertrophy(AH)with allergic rhinitis(AR)in the otitis media with effusion(OME)in children,as well as to discuss the risk factors on the inducement of OME in patients with AH.[Methods]The clinical materials were collected and analyzed from 205 children with AH who admitted for surgical treatment between 2013 and 2015, including medical history and signs,acoustic immittance measurement,allergy screen as well as blood routine,and to evaluate the situation of OME and AR in patients with AH. All the data were analyzed by SPSS 20.0.[Results]Among the 205 AH children,66 cases(32.20%)were accompanying with OME. The AH patients aged 3-5 years had the highest incidence of OME,which decreased with age(P=0.018). It is significant on the prevalence of the OME in patients with AH between the two groups with and without AR (P = 0.010). In the logistic model investigating the risk factors of inducing OME among patients with AH,the third-degree AH, Type-three and the accompanying with AR were significant. The third-degree and third-type AH children were 2.729 and 6.390-folds higher than others respectively (P = 0.047,P = 0.001). The incidence of OME appeared to be 1.212-folds higher among patients with AR(P = 0.010).[Conclusion]Mechanical obstruction and AR played a superimposed role in the inducement of OME among patients with AH,which was effected by multiple factors. We should pay attention to the“lateral respiratory”allergic diseases on eustachian tube and middle ear.

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae ; Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; Transfection