Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus(HBV). Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northem blot, and strong signals could be seen from day 7 to day 14 post-in-fection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably repli-cate and express in HBV-infected tupaia hepatocytes.

The 68 Center for Drug Evaluation recommend approved innovative drugs between 2019 and 2021 evaluation situation was summarized in this article,to analyzes the innovative drugs research and development trend and the influence of the drug evaluation and approval system reform.Meanwhile,after sorting out the clinical trials inspection report,to analyze the clinical trials on-site inspection of innovative drugs,and summarized the common defects,hope to provide reference and suggestions for subsequent innovative drug application and high-quality drug clinical trials.

Objective To investigate the value of non-contrast CT(NCCT)-based radiomics for identifying acute unilateral middle cerebral artery occlusion(MCAO)with negative hyperdense artery sign(HAS).Methods All 80 patients with acute unilateral MCAO confirmed by angiography(MR angiography[MRA]or CT angiography[CTA]or DSA)and presenting with negative NCCT presentation for HAS were enrolled from January 2015 to June 2023 in the Emergency Department of Stroke Center of Affiliated Hospital of Yangzhou university.On the NCCT images,the occluded segment of the middle cerebral artery on the affected side of each case and the corresponding segment of the vessel on the normal side were used as the regions of interest,and a total of 108 radiomic features were extracted.The least absolute shrinkage and selection operator(LASSO)was used to screen the key features,construct and calculate the radiomics score,and four imaging histology models,support vector machine(SVM),light gradient boosting machine(LightGBM),GradientBoosting and adaptive boosting(AdaBoost),were built respectively to predict MCAO.Predictive performance was evaluated by the area under the receiver operating characteristic curves,and comparisons between the modeled receiver operating characteristic curves were made using the Delong test.Finally,the value of the application of radiological modeling was assessed by clinical decision curve analysis(DCA).Results The NCCT images based on 160 vessels were finally screened for 6 key features,including skewness,energy,gray level size zone matrix(GLSZM)-gray uneven,GLSZM-low gray area emphasis,GLSZM-size area non-uniform standardization,GLSZM-area entropy.The area under the curve(AUC)of the SVM-test was 0.688(95％CI 0.497-0.878)with an accuracy of 0.688;the AUC of the LightGBM-test was 0.787(95％CI 0.620-0.955)with an accuracy of 0.781;the AUC of the GradientBoosting-test was 0.654(95％CI 0.457-0.852)with an accuracy of 0.688;the AUC of the AdaBoost-test was 0.707(95％CI 0.515-0.899)with an accuracy of 0.750.The Delong test showed a statistically significant difference between LightGBM-test and GradientBoosting-test(P=0.040),and no statistically significant difference in performance between the remaining models(all P＞0.05).DCA showed that the LightGBM-test performed better.Conclusion NCCT-based radiomics has good diagnostic efficacy for identifying acute unilateral MCAO with negative HAS,and this conclusion needs to be further verified by multi-center and large sample studies.

Objective To assess the clinical efficacy of butylphthalide sequential therapy(BST)as an adjuvant therapy for delayed onset encephalopathy caused by acute carbon monoxide poisoning(DEACMP).Methods DEACMP patients who visited the department of neurology at Yan'an People's Hospital from January 2018 to June 2023 were selected as the research subjects.Using a random number table method,DEACMP patients were randomly divided into hyperbaric oxygen therapy(HBOT)group,glucocorticoid(GC)group(GC combined with,HBOT),and BST group(HBOT,GC combined with BST),all of which were treated continuously for 30 days.The incidence of adverse reactions and clinical efficacy during the treatment among three groups of DEACMP were observed and compared.Inflammation indicators,neurological damage indicators,cognitive function,neurological function,and daily living ability before and after treatment among three groups were compared.Results A total of 120 patients were included in the study,with 40 patients in each group.During the treatment period,no serious adverse reactions were observed in any of the three groups of patients,and no patient terminated treatment due to adverse reactions.The total effective rate of BST group was significantly higher than those of GC group and HBOT group,while the total effective rate of GC group was higher than that of HBOT group(P<0.05).Before treatment,there was no statistically significant difference(P>0.05)in the scores of inflammation indicators,neurological damage indicators,cognitive function,neurological function,and daily living function among the three groups.After treatment,the serum LP-PLA2,SIRI,TNF-α,S100-β,and the NSE levels decreased compared to before treatment(P<0.05),and the indicators of the BST group was lower than those of the HBOT group and GC group,while the indicators of the GC group was lower than those of the HBOT group(P<0.05).After treatment,the MMSE,MoCA,and BI scores of the three groups increased compared to before treatment(P<0.05),but the NIHSS score decreased(P<0.05).In addition,the MMSE,MoCA,and BI scores of the BST group were higher than those of the HBOT and GC groups(P<0.05),while the NIHSS scores were lower than those of the HBOT and GC groups(P<0.05).The MMSE,MoCA,and BI scores in the GC group were higher than those in the HBOT group,while the NIHSS scores were lower than those in the HBOT group(P<0.05).Conclusion Compared with HBOT treatment and HBOT combined with GC treatment,BST adjuvant therapy can improve the clinical efficacy,inflammatory status,cognitive function,and daily living ability of DECAMP patients.

Objective To study the protective effect of Irisin in doxorubicin(Dox)induced-Cardiomyopathy and its possible mechanism.Methods AC 16 cells were used to construct Dox injury model and divided into control group(AC 16 cells were cultured with complete medium),Irisin group(AC16 cells were treated with 10 ng·L-1 Irisin for 24 h),Dox group(AC 16 cells were treated with 4 μmol·L-1 Dox for 24 h),Dox+Irisin group(AC 16 cells were pretreated with 10 ng·L-1 Irisin for 2 h,and then treated with 4 pmol·L-1 Dox for 24 h).Cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)and lactate dehydrogenase(LDH)were used to detect the proliferation,apoptosis and mortality of AC 16 cells.Western blot was used to detect the expression levels of nuclear factor-κB(NF-κB)signaling pathway and apoptotic factors B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-9 protein.Mito-Tracker Red CMXRos probe was used to detect mitochondrial membrane potential.Results In the contrl group,Irisin group,Dox group,Dox+Irisin group,the rate of apoptosis were(0.97±0.09)％,0,(42.80±6.70)％,(11.74±1.79)％;the expression of Bax protein were 0.85±0.01,0.36±0.02,1.15±0.07,0.37±0.11;the expression of caspase-9 protein were 0.52±0.02,0.59±0.03,1.11±0.02,0.67±0.08;the expression of Bcl-2 protein were 1.01±0.04,1.05±0.25,0.43±0.02 and 0.99±0.30;the probability of mitochondrial damage were(0.02±0.01)％,(0.5±0.15)％,(38.6±2.39)％,(1.58±0.54)％.The difference of the above indexes between the contrl group and the Dox group were statistically significant(all P＜0.05);the difference between Dox group and Dox+Irisin group were statisically significant(all P＜0.05).Conclusion Irisin could reduce the expression level of Bax,caspase-9,p-NF-κB,and p-mTOR caused by Dox,increase the expression level of Bcl-2,ameliorate the myocardial damage caused by Dox,and reduce cardiotoxicity.

BACKGROUNDVanishing white matter disease (VWM), a human autosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B (eIF2B). eIF2B is responsible for the initiation of protein synthesis by its guanine nucleotide exchange factor (GEF) activity. Mutations of eIF2B impair GEF activity at different degree. Previous studies implied improperly activated unfolded protein response (UPR) and endoplasmic reticulum stress (ERS) participated in the pathogenesis of VWM. Autophagy relieves endoplasmic reticulum load by eliminating the unfolded protein. It is still unknown the effects of genotypes on the pathogenesis. In this work, UPR and autophagy flux were analyzed with different mutational types.

METHODSERS tolerance, reflected by apoptosis and cell viability, was detected in human oligodendrocyte cell line transfected with the wild type, or different mutations of p. Arg113His, p. Arg269FNx01 or p. Ser610-Asp613del in eIF2Bε. A representative UPR-PERK component of activating transcription factor 4 (ATF4) was measured under the basal condition and ERS induction. Autophagy was analyzed the flux in the presence of lysosomal inhibitors.

RESULTSThe degree of ERS tolerance varied in different genotypes. The truncated or deletion mutant showed prominent apoptosis cell viability declination after ERS induction. The most seriously damaged GEF activity of p. Arg269FNx01 group underwent spontaneous apoptosis. The truncated or deletion mutant showed elevated ATF4 under basal as well as ERS condition. Decreased expression of LC3-I and LC3-II in the mutants reflected an impaired autophagy flux, which was more obvious in the truncated or deletion mutants after ERS induction.

CONCLUSIONSGEF activities in different genotypes could influence the cell ERS tolerance as well as compensatory pathways of UPR and autophagy. Oligodendrocytes with truncated or deletion mutants showed less tolerable to ERS.

Cell Line ; Endoplasmic Reticulum Stress ; genetics ; physiology ; Eukaryotic Initiation Factor-2B ; genetics ; Humans ; Mutation ; genetics ; Oligodendroglia ; metabolism ; Unfolded Protein Response ; genetics ; physiology

Objective:To understand the current state of ethical competence levels of nurses and analyse the factors that influence them, in order to inform the development of targeted training programmes.Methods:This study adopted a cross-sectional survey method and used convenience sampling to select 825 clinical nurses from Tianjin Chest Hospital as the survey objects from July to August, 2023, and a questionnaire survey was conducted using the General Information Questionnaire, the Ethical Competence Scale, Ethical Competence Support Scale, and Ethical Safety Scale.Results:A total of 818 valid questionnaires were retracted. Among the 818 nurses, 48 were males and 770 were females, the age was (33.19 ± 7.40) years. The total score of nurses ′ Ethical Competence, Ethical Competence Support and Ethical Safety were (118.08 ± 19.96), (215.07 ± 32.02), (48.93 ± 7.55) points, all of which were at a high level. The total score of nurses ′ ethical competence were positively correlated with ethical competence support and ethical safety ( r=0.856, 0.830, both P<0.01); multivariate linear regression analysis showed that the department, the level of ethical competence support, and the level of ethical safety were the influencing factors of the level of ethical competence ( t=5.19, 12.35, 3.88, all P<0.01). Conclusions:Nurses ′ ethical competence, ethical competence support, and ethical safety were at a high level, and the department, the level of ethical competence support, and the level of ethical safety were the factors influencing the level of ethical competence. Nursing managers can provide more ethical education and address ethical issues in multiprofessional discussions, strengthen organizational and personal support for nurses ′ ethical competence, improve nurses ′ ethical safety, and help them implement good ethical care.

Background and objective: Coronavirus Disease 2019 (COVID19) was first reported in Malaysia in March 2020. We describe here the clinical characteristics and computed tomography (CT) patterns in asymptomatic young patients who had laboratory-confirmed COVID-19. Methods: This is a retrospective observational study where 25 male in-patients with laboratory-confirmed COVID-19 in Hospital Canselor Tuanku Muhriz. Demographics, clinical data and CT images of these patients were reviewed by 2 senior radiologists. Results: In total there were 25 patients (all males; mean age [±SD], 21.64±2.40 years; range, 18-27 years). Patients with abnormal chest CT showed a relatively low normal absolute lymphocytes count (median: 2.2 x 109/L) and absolute monocyte count (median: 0.5 x 109/L). Lactate dehydrogenase was elevated in 5 (20%) of the patients. The procalcitonin level was normal while elevated levels of alanine aminotransferase, total bilirubin, platelet and C-reactive protein were common. Baseline chest CT showed abnormalities in 6 patients. The distribution of the lesions were; upper lobe 3 (12%) lower lobe 3 (12%) with peripheral distribution 4 (16%). Of the 25 patients included, 4 (16%) had ground glass opacification (GGO), 1 (4%) had a small peripheral subpleural nodule, and 1 (4%) had a dense solitary granuloma. Four patients had typical CT features of COVID-19. Conclusion: We found that the CT imaging showed peripheral GGO in our patients. They remained clinically stable with no deterioration of their respiratory symptoms suggesting stability in lung involvement. We postulate that rapid changes in CT imaging may not be present in young, asymptomatic, non-smoking COVID-19 patients. Thus the use of CT thorax for early diagnosis may be reserved for patients in the older age groups, and not in younger patients.