Objective To investigate the molecular mechanism of Brusatol(BRU)regulating apoptosis of Hela cells through TLR9-MyD88 signaling pathway.Methods Hela cells preserved in our laboratory were treated with Brucea javanica at different concentrations(0,5.0,10.0,20.0,40.0,80.0 nmol·L-1).Hela cells were divided into Control group(normal cultured Hela cells),BRU-L group(treated with 10.0 nmol·L-1 Brucea javanica)and BRU group-H(treated with 20.0 nmol·L-1 Brucea javanica)Cells were treated with nmol·L-1 of Brucea javanica),BRU+pcDNA-NC group(transfected with pcDNA-NC+20.0 nmol·L-1 of Brucea javanica),BRU-H+pcDNA-TLR9 group(transfected with pcDNA-TLR9+20.0 nmol·L-1 of Brucea javanica).Cell proliferation was detected by CCK-8 and EdU methods.Apoptosis was detected by TUNNEL staining and flow cytometry.The protein expression levels of TLR9,MyD88,Bax,Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were detected by Western blot.Cell apoptosis and mitochondrial membrane potential detection kit(JC-1)were detected by flow cytometry,and the contents of adenosine triphosphate(ATP)and reactive oxygen species(ROS)were detected by ELISA.Results Compared with 0 nmol·L-1 group,the survival rate and IC50 value of 10 nmol·L-1 group were significantly decreased(P<0.05).After stimulation of BRU with different concentrations,the proliferation ability of cells was significantly decreased in a dose-dependent manner(P<0.05).Compared with control group,the 5-ethynyl-2'-deoxyuracil(EDU)positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein of cells in BRU-L,BRU-H and pcDNA-NC groups were significantly decreased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly increased(P<0.05).In control group,BRU-L,BRU-H group/BRU-H+pcDNA-NC,there was a continuous decreasing trend(P<0.05).Compared with the BRU-H+pcDNA-NC group,the EDU positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein in the BRU-H+pcDNA-TLR9 group were significantly increased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly decreased(P<0.05).Compared with Control group,JC-1 level and ATP content in BRU-L,BRU-H and BRU-H+pcDNA-NC groups were significantly decreased,while ROC content and mitotracker positive cell level were significantly increased(P<0.05).Compared with BRU-L group,JC-1 level and ATP content in BRU-H group and BRU-H+pcDNA-NC group were further decreased,while ROC content and mitotracker positive cell level were further increased(P<0.05).Compared with the BRU-H+pcDNA-NC group,the levels of JC-1 and ATP in the BRU-H+pcDNA-TLR9 group were increased,while the levels of ROC,mitotracker staining positive cells were decreased(P<0.05).Conclusion Brucea javanica can produce Hela cell proliferation by regulating TLR9-MyD88 signaling pathway.

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

Objective::This study aimed to estimate the proportion of non-obstructive coronary artery disease (CAD) patients in large percutaneous coronary intervention (PCI) centers in China.Methods::The study was conducted at 6 large PCI centers in China from January 1, 2013 to December 31, 2015. Demographic and clinical data were collected from medical records, prescription records, and laboratory reports of patients with symptoms of angina who underwent coronary angiography (CAG).Results::A total of 1713 patients were consecutively screened, 1600 of whom were included in the study. CAG showed that 300 patients had non-obstructive CAD while 1300 had obstructive CAD. Among the 300 patients with non-obstructive CAD, 203 displayed mild coronary stenosis (20%-49%) and 91 had normal coronary status (ie, <20% stenosis). Of the 1300 patients with obstructive CAD, 61.6% (801/1300) had typical symptoms of angina, compared with 49.3% (148/300) for patients with non-obstructive CAD. In addition, there were more women than men in the non-obstructive CAD group, whereas the reverse was observed among obstructive CAD patients.Conclusion::The prevalence of non-obstructive CAD in the Chinese population with coronary heart disease was estimated to be approximately 20%. Additionally, typical angina symptoms were correlated with obstructive CAD, whereas female gender was identified as a risk factor for non-obstructive CAD.

Objective::This study aimed to estimate the proportion of non-obstructive coronary artery disease (CAD) patients in large percutaneous coronary intervention (PCI) centers in China.Methods::The study was conducted at 6 large PCI centers in China from January 1, 2013 to December 31, 2015. Demographic and clinical data were collected from medical records, prescription records, and laboratory reports of patients with symptoms of angina who underwent coronary angiography (CAG).Results::A total of 1713 patients were consecutively screened, 1600 of whom were included in the study. CAG showed that 300 patients had non-obstructive CAD while 1300 had obstructive CAD. Among the 300 patients with non-obstructive CAD, 203 displayed mild coronary stenosis (20%-49%) and 91 had normal coronary status (ie, <20% stenosis). Of the 1300 patients with obstructive CAD, 61.6% (801/1300) had typical symptoms of angina, compared with 49.3% (148/300) for patients with non-obstructive CAD. In addition, there were more women than men in the non-obstructive CAD group, whereas the reverse was observed among obstructive CAD patients.Conclusion::The prevalence of non-obstructive CAD in the Chinese population with coronary heart disease was estimated to be approximately 20%. Additionally, typical angina symptoms were correlated with obstructive CAD, whereas female gender was identified as a risk factor for non-obstructive CAD.

Objective: To analyle prevalence and trend of substance abuse behaviors among adolescents in Guangdong province and to provide theoretical evidence for effective interventions. Methods: Data regarding smoking, drinking, drugs abuse from Adolescents Health Risk Behavior Survey in 2007, 2010, 2013 and 2016 were collected and comparatively analyzed. Results: The incidences of tried smoking(27.37%), current smoking(8.32%), passive smoking(45.52%), current drinking(27.97%) and drug using(0.75%) in 2016 were lower than that in 2007(32.40%, 10.81%, 52.69%, 31.97%, 1.35%), and the average annual decrease rate were 1.89%, 2.95%, 1.64%, 1.50%, 6.75%, respectively(χ2=152.73, 92.50, 313.92, 46.48, 46.64, P<0.01). During the past decate, the incidences of current smoking in rural adolescents(12.02%, 10.64%, 9.99%, 10.10%) were significantly higher than that in urban adolescents(8.88%, 8.52%, 8.17%, 6.56%)(χ2=54.46, 26.62, 25.23, 100.43, P<0.01); the incidences of current drinking in urban adolescents(33.57%, 30.37%, 35.76%, 30.46%) were significantly higher than that in rural adolescents(30.97%, 28.48%, 28.47%, 25.32%)(χ2=16.71, 10.13, 153.47, 81.76, P<0.01); the incidence of current drinking was the highest among college students in each monitoring year(47.71%, 41.76%, 42.36%, 37.77%)(χ2=583.37, 681.25, 610.04, 729.02, P<0.01); the incidence of current smoking was the highest among vocational school students in each monitoring year(16.36%, 17.00%, 14.90%, 16.71%)(χ2=256.45, 515.61, 339.63, 742.52, P<0.01); In addition to the incidence of drug abuse was the highest among college students in 2010(2.02%), vocational school students were the highest in other years(χ2=66.92, 34.38, 23.08, 20.55, P<0.01). Conclusion Smoking and drug abuse among adolescents in Guangdong showed a declining trend by years, as well as the prevalence of drinking behavior. Strengthened intervention should be targeted on smoking behavior among rural students, drinking behavior among urban students, and smoking behavior and drug abuse among vocational school students.

Objective To determine the expression of disabled homolog 2 interacting protein (DAB2IP) gene in the basal cell carcinoma (BCC) of the skin,and to investigate its clinical significance.Methods Clinical data were retrospectively analyzed in 105 outpatients and inpatients who received skin mass resection in Department of Dermatology,Guangdong Second Provincial General Hospital and Guangzhou Institute of Dermatology between January 2012 and November 2017.Totally,79 patients with pathologically diagnosed BCC of the skin served as patient group,and 26 patients with pathologically diagnosed skin tag but without other clinical manifestations served as control group.Immunohistochemical staining was performed to determine the expression of DAB2IP in the two groups,and correlations of the DAB2IP expression with the clinical phenotype and pathological features of BCC of the skin were analyzed.Statistical analysis was carried out with SPSS21.0 software by using chi-square test for the comparison of enumeration data.Results The protein expression of DAB2IP was observed in 11 (42.3％) of 26 patients in the control group,as well as in 74 (93.7％) of 79 patients in the patient group,and there was a significant difference in the positive rate of DAB2IP protein between the two groups (x2 =33.50,P ＜ 0.05).The expression of DAB2IP was uncorrelated with gender or age of patients with BCC of the skin,or with the tumor size (all P ＞ 0.05).The positive rate of DAB2IP protein significantly differed between the patients with superficial BCC (5/7) and those with invasive BCC (95.8％,69/72;x2 =6.47,P ＜ 0.05).Of the 79 patients with BCC of the skin,Ki-67 protein was detected in 31 (39.2％),and the cancer cells expressing Ki-67 protein also expressed DAB2IP protein.Conclusion The expression of DAB2IP increases in BCC of the skin,which may be associated with the occurrence and infiltration of BCC of the skin.

Objective To investigate the change of the error positivity(Pe)of event-related poten-tial(ERP)under Flanker task in adolescents with smoking addiction. Method The signals of ERP during the Flanker task in fifteen adolescents with smoking addiction were compared with that of 15 non-smoking ad-olescents. Results The Pe amplitude in adolescents with smoking addiction((3.575±2.299)μV)was sig-nificantly lower than that of non-smokers((5.865 ± 4.290)μV)(F=5.324,P=0.029),and the reaction time of the Flanker task in smoking addiction adolescent group((440.754± 36.051)ms)was significantly higher than that in the control group((422.850±20.194)ms)(F=0.620,P<0.05).The Pe amplitude in ado-lescents with smoking addiction was positively correlated with smoking index(pack-years)(r=0.599,P=0.018). Conclusion The adolescents with smoking addiction may exit worse ability of error monitoring dur-ing the Flanker task.

Objective To observe the changes of electroencephalogram (EEG) signal during the resting state in adolescents with smoking addiction.Methods Spectral analysis during the resting state was used to observe and analyze the EEG signals between adolescents with smoking addiction (n=21) and nosmoking control group (n=21) by using Brain Vision Analyzer software.Results Compared with the nosmoking control group,adolescents with smoking addiction showed increased alpha band signals in 3 brain regions (including left-anterior,right-anterior and midline-anterior regions,P＜0.05),and decreased delta band signals in 3 brain regions (including right-posterior,midline-anterior and midline-posterior regions,P＜0.05).Moreover,the signals in these brain regions were positively correlated with pack-years (alpha band:left-anterior(r=0.648,P=0.003),right-anterior (r=0.629,P=0.004),midline-anterior (r=0.456,P=0.043);delta band:right-posterior (r=0.602,P =0.005),midline-anterior (r =0.533,P=0.019),midline-posterior (r=0.692,P=0.001))in adolescents with smoking addiction.Conclusion The changes of EEG signals during the resting state in adolescents with smoking addiction may be associated with cumulative effects of smoking.

Objective To study the psychological status of patients having undergone eyeball enucleation,and put forward the corresponding strategy for psychological nursing.Method The symptom checklist-90(SCL-90)was used to investigate their psychological status after eyeball enucleation.Result The scores on interpersonal relationship sensitivity factor,depression factor, anxiety factor and terror factor(28?78±0?61),(40?76±0?65),(33?86±0?62)and(23?88±0?46),respectively.Conclusions Pertinent psychological nursing may help patients to eliminate the adverse emotional and psychological pressures,promoting them to develop a good state of mind,and return to the society better?

Objective To compare the characterization and myocardial differentiation capacity of amniotic fluid-derived mesenchymal stromal cells (AF MSCs) and umbilical cord Wharton's Jelly-derived mesenchymal stromal cells (WJ MSCs). Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton's Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes (GATA-4, c-TnT, α-actin, and Cx43) were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities of WJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility (MHC I) antigens (HLA-ABC), and were negative, or mildly positive, for MHC Class II (HLA-DR) antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, α-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.