Objective Genome structural equation modeling(GSEM)was used to study the shared genetic structure between Alzheimer's disease(AD)and eight cardiovascular diseases(CVD).Methods In this study,based on AD and CVD data from large consortia and genome-wide association studies,we used chained unbalanced score regression to assess the genetic correlation between the diseases;GSEM was introduced to cross-validate with exploratory factor analysis at odd chromosomes and validation factor analysis at even chromosomes,and pathway analysis was combined to establish three-factor,cofactor,and bifactorial models,so as to reveal the complex genetic relationships between the diseases.Additionally,a cross-validation was performed by swapping the positions of odd and even chromosomes.Results The nine selected diseases can be explained by a three-factor model consisting of three independent factors;a cofactor model explains the relationship between a cofactor and each disease;and an extended two-factor model(a combination of the three-factor model and the cofactor model)performs best in fitting the study data.Conclusion GSEM can be used to explore the shared genetic structure between AD and CVD to discover their interactions,laying a methodological foundation for understanding the genetic relationship between complex diseases.

Objective The Bayesian joint model was applied to predict conversion from mild cognitive impairment(MCI)to Alzheimer's disease(AD),and to compare time effect in longitudinal outcomes,and functional forms for the longitudinal outcomes that are included in the survival model.Methods The longitudinal sub-model used generalized linear mixed models to model the trajectory of longitudinal neuropsychological tests changes pattern,and the survival sub-model adopted a Cox proportional risk model.The longitudinal sub-model utilized two forms of effects,t and t2,and the link structures of longitudinal sub-model and survival sub-model adopted three functional forms(updated value,slope,and area).The data were divided into training and validation sets according to 7∶3,the model parameters were estimated based on Bayesian algorithm,and the dynamic AUC was used to evaluate model prediction performance.Results There were 374(54.20％)of 690 patients with MCI progressed to AD during the follow-up period.In training set,the results of Bayesian joint model with a time effect of t and area effect of three longitudinal neuropsychological tests were optimal,with a mean dynamic AUC of 0.8336.The APOEε4 gene and a low functional activities questionary were risk factors for MCI conversion to AD.The dynamic AUCs of the joint model with the t time effects and area effect for longitudinal data were above 0.75 in the validation sets.Conclusion The application of multivariate Bayesian joint model to MCI risk prediction may provide a theoretical basis for individualized interventions for cognitive impairment and inform statistical methods for survival modeling of chronic diseases.

Objective Genome structural equation modeling(GSEM)was used to study the shared genetic structure between Alzheimer's disease(AD)and eight cardiovascular diseases(CVD).Methods In this study,based on AD and CVD data from large consortia and genome-wide association studies,we used chained unbalanced score regression to assess the genetic correlation between the diseases;GSEM was introduced to cross-validate with exploratory factor analysis at odd chromosomes and validation factor analysis at even chromosomes,and pathway analysis was combined to establish three-factor,cofactor,and bifactorial models,so as to reveal the complex genetic relationships between the diseases.Additionally,a cross-validation was performed by swapping the positions of odd and even chromosomes.Results The nine selected diseases can be explained by a three-factor model consisting of three independent factors;a cofactor model explains the relationship between a cofactor and each disease;and an extended two-factor model(a combination of the three-factor model and the cofactor model)performs best in fitting the study data.Conclusion GSEM can be used to explore the shared genetic structure between AD and CVD to discover their interactions,laying a methodological foundation for understanding the genetic relationship between complex diseases.

Objective The Bayesian joint model was applied to predict conversion from mild cognitive impairment(MCI)to Alzheimer's disease(AD),and to compare time effect in longitudinal outcomes,and functional forms for the longitudinal outcomes that are included in the survival model.Methods The longitudinal sub-model used generalized linear mixed models to model the trajectory of longitudinal neuropsychological tests changes pattern,and the survival sub-model adopted a Cox proportional risk model.The longitudinal sub-model utilized two forms of effects,t and t2,and the link structures of longitudinal sub-model and survival sub-model adopted three functional forms(updated value,slope,and area).The data were divided into training and validation sets according to 7∶3,the model parameters were estimated based on Bayesian algorithm,and the dynamic AUC was used to evaluate model prediction performance.Results There were 374(54.20％)of 690 patients with MCI progressed to AD during the follow-up period.In training set,the results of Bayesian joint model with a time effect of t and area effect of three longitudinal neuropsychological tests were optimal,with a mean dynamic AUC of 0.8336.The APOEε4 gene and a low functional activities questionary were risk factors for MCI conversion to AD.The dynamic AUCs of the joint model with the t time effects and area effect for longitudinal data were above 0.75 in the validation sets.Conclusion The application of multivariate Bayesian joint model to MCI risk prediction may provide a theoretical basis for individualized interventions for cognitive impairment and inform statistical methods for survival modeling of chronic diseases.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective:To investigate the effect of tofacitinib,a pan-Janus kinase(JAK)inhibitor,on transforming growth factor-beta 1(TGF-β1)-induced fibroblast to myofibroblast transition(FMT)and to explore its mechanism.To provide a theoretical basis for the clinical treatment of connective tissue disease-related interstitial lung disease(CTD-ILD).Methods:(1)Human fetal lung fibroblast 1(HFL-1)were cultured in vitro,and 6 groups were established:DMSO blank control group,TGF-β1 in-duction group,and TGF-β1 with different concentrations of tofacitinib(0.5,1.0,2.0,5.0 μmol/L)drug intervention experimental groups.CCK-8 was used to measure the cell viability,and wound-healing assay was performed to measure cell migration ability.After 48 h of combined treatment,quantitative real-time PCR(RT-PCR)and Western blotting were used to detect the gene and protein expression levels of α-smooth muscle actin(α-SMA),fibronectin(FN),and collagen type Ⅰ(COL1).(2)RT-PCR and enzyme-linked immunosorbnent assay(ELISA)were used to detect the interleukin-6(IL-6)gene and protein expression changes,respectively.(3)DMSO carrier controls,1.0 μmol/L and 5.0 μmol/L tofacitinib were added to the cell culture media of different groups for pre-incubation for 30 min,and then TGF-β1 was added to treat for 1 h,6 h and 24 h.The phosphorylation levels of Smad2/3 and signal transducer and activator of transcription 3(STAT3)protein were detected by Western blotting.Results:(1)Tofacitinib inhibited the viability and migration ability of HFL-1 cells after TGF-β1 induction.(2)The expression of α-SMA,COL1A1 and FN1 genes of HFL-1 in the TGF-β1-induced groups was signifi-cantly up-regulated compared with the blank control group(P＜0.05).Compared with the TGF-β1 in-duction group,α-SMA expression in the 5.0 μmol/L tofacitinib intervention group was significantly inhi-bited(P＜0.05).Compared with the TGF-β1-induced group,FN1 gene was significantly inhibited in each intervention group at a concentration of 0.5-5.0 μmol/L(P＜0.05).Compared with the TGF-β1-induced group,the COL1A1 gene expression in each intervention group did not change significantly.(3)Western blotting results showed that the protein levels of α-SMA and FN1 in the TGF-β1-induced group were significantly higher than those in the control group(P＜0.05),and there was no significant difference in the expression of COL1A1.Compared with the TGF-β1-induced group,the α-SMA protein level in the intervention groups with different concentrations decreased.And the differences between the TGF-β1-induced group and 2.0 μmol/L or 5.0 μmol/L intervention groups were statistically significant(P＜0.05).Compared with the TGF-β1-induced group,the FN1 protein levels in the intervention groups with different concentrations showed a downward trend,but the difference was not statistically sig-nificant.There was no difference in COL1A1 protein expression between the intervention groups com-pared with the TGF-β1-induced group.(4)After TGF-β1 acted on HFL-1 cells for 48 h,the gene ex-pression of the IL-6 was up-regulated and IL-6 in culture supernatant was increased,the intervention with tofacitinib partly inhibited the TGF-β1-induced IL-6 gene expression and IL-6 in culture supernatant.TGF-β1 induced the increase of Smad2/3 protein phosphorylation in HFL-1 cells for 1 h and 6 h,STAT3 protein phosphorylation increased at 1 h,6 h and 24 h,the pre-intervention with tofacitinib inhibited the TGF-β1-induced Smad2/3 phosphorylation at 6 h and inhibited TGF-β1-induced STAT3 phosphorylation at 1 h,6 h and 24 h.Conclusion:Tofacitinib can inhibit the transformation of HFL-1 cells into myofi-broblasts induced by TGF-β1,and the mechanism may be through inhibiting the classic Smad2/3 path-way as well as the phosphorylation of STAT3 induced by TGF-β1,thereby protecting the disease progres-sion of pulmonary fibrosis.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Photothermal therapy has been intensively investigated for treating cancer in recent years. However, the long-term therapeutic outcome remains unsatisfying due to the frequently occurred metastasis and recurrence. To address this challenge, immunotherapy has been combined with photothermal therapy to activate anti-tumor immunity and relieve the immunosuppressive microenvironment within tumor sites. Here, we engineered silica-based core‒shell nanoparticles (JQ-1@PSNs-R), in which silica cores were coated with the photothermal agent polydopamine, and a bromodomain-containing protein 4 (BRD4) inhibitor JQ-1 was loaded in the polydopamine layer to combine photothermal and immune therapy for tumor elimination. Importantly, to improve the therapeutic effect, we increased the surface roughness of the nanoparticles by hydrofluoric acid (HF) etching during the fabrication process, and found that the internalization of JQ-1@PSNs-R was significantly improved, leading to a strengthened photothermal killing effect as well as the increased intracellular delivery of JQ-1. In the animal studies, the multifunctional nanoparticles with rough surfaces effectively eradicated melanoma via photothermal therapy, successfully activated tumor-specific immune responses against residual tumor cells, and further prevented tumor metastasis and recurrence. Our results indicated that JQ-1@PSNs-R could serve as an innovative and effective strategy for combined cancer therapy.

Objective:To investigate epidemiological characteristics and macrolide-resistance of hospitalized children with Mycoplasma pneumoniae (MP) infections in Beijing from 2016 to 2019, so as to provide basis for the prevention and treatment of pediatric Mycoplasma pneumoniae pneumonia (MPP).Methods:The clinical data were analyzed retrospectively from 8 691 children hospitalized with community acquired pneumonia in Beijing Children′s Hospital between January 2016 and September 2019.MP RNA was detected by simultaneous amplification and testing (SAT), and macrolide resistance of MP was examined by MP and macrolide-resistant isolate diagnostic kit (PCR with fluorescence probes). Chi- square test was used for categorical analysis. Results:Among 8 691 cases detected by SAT, the overall detection rate of MP was 28.10% (2 442/8 691 cases). The detection rates of MP from 2016 to 2019 were 26.23%, 31.36%, 27.84 % and 26.57%, respectively.The detection rate of MP in 2017 was significantly higher than that in other years ( χ2=16.11, P<0.05). The detection rate of MP in females was 29.65%(1 107/3 733 cases), which was evidently higher than that in males 26.93%(1 335/4 958 cases) ( χ2=7.85, P<0.05). The positive rates of MP in summer[32.21% (726/2 254 cases)] and autumn[39.76%(852/2 143 cases)] were significantly higher than those in spring[17.00% (327/1 924 cases)] and winter[22.66%(537/2 370 cases)] ( χ2=315.15, P<0.001). The percentages of MP were 35.06%(732/2 088 cases) in preschoolers and 37.71%(1 160/3 076 cases) in school-age children, which were significantly higher than 11.20%(232/2 072 cases) in infants and 22.01% (318/1 445 cases) in toddlers ( χ2=509.89, P<0.001). Macrolide resistance detection was conducted in 1 524 patients by fluorescent PCR.Among them, 1 386 patients were positive for drug resistance, and the positive rate was 90.94%.The prevalence of macrolide-resistant MP from 2016 to 2019 were 88.19%, 90.93%, 90.56% and 92.90%, respectively.Macrolide-resistant rates were not related with gender, age and season. Conclusions:MP can be detected in all seasons, but most prevalently in summer and autumn.Girls are more prone to MP infections than boys.The detection rate of MP increases with age, and the positive rate is higher in preschoolers and school-age children.During the 4-year study period, the drug resistant rate of MP remain high.

Objective To investigate the current status of self-perceived burden and mindfulness of chronic obstructive pulmonary disease (COPD) patients and to explore the effect of mindfulness on patients' self-perceived burden. Methods From October 2016 to April 2017, a total of 276 patients with COPD were selected from a ClassⅢ Grade A hospital in Harbin by convenience sampling. The general information questionnaire, Five Facet Mindfulness Questionnaire (FFMQ), Self-Perceived Burden Scale (SPBS) were used to investigate the patients. Pearson correlation and multiple linear regression were used to analyze the effect of mindfulness on self-perceived burden. Results The score of self-perceived burden of COPD patients was (33.34±6.54) with a medium level. The score of mindfulness was (129.87±25.78). Pearson correlation showed the scores of dimensions and the total score of mindfulness except for the dimension of observation among COPD patients were negatively correlated with the self-perceived burden (P<0.05). Multiple linear regression revealed that three dimensions, mindfulness description, mindfulness action and no judging, entered the regression equation which were the influencing factors of self-perceived burden (P<0.05). Conclusions COPD patients commonly have self-perceived burden with different degrees which influenced by mindfulness. Health care providers could improve patients' mindfulness by reasonable intervention so as to reduce their self-perceived burden.