Objective To investigate the mechanism of morin-induced autophagy in non-small cell lung cancer A549 cells based on mTOR/STAT3 signaling axis.Methods A549 cells were divided into blank group and 30,60,90,120 and 150 μg·mL-1 of morin groups.After 24,48 and 72 hours of culture,the cell proliferation activity was detected by CCK-8 method,and the cell inhibition rate was calculated.A549 cells were divided into blank group and 30,90,150 μg·mL-1 morin groups.After 14 days of culture,the cell proliferation was detected by colony formation assay.After 24 hours of culture,the cell proliferation ability was detected by BeyoClickTM EdU-488.Apoptosis was detected by flow cytometry;acridine orange staining was used to detect cell autophagy;the formation of autophagosomes was observed by transmission electron microscopy.Western Blot was used to detect the expression levels of apoptosis,autophagy and mTOR/STAT3 signaling axis-related proteins in cells.A549 cells were divided into blank group,blank group + chloroquine(10 μg·mL-1)group,morin(30,150 μg·mL-1)group,morin(30,150 μg·mL-1)+ chloroquine(10 μg·mL-1)group.After 48 hours of intervention,the cell activity was detected by CCK-8 method,and the cell survival rate was calculated.Results Compared with the blank group,the inhibition rate of A549 cells in 60,90,120,150 μ g·mL-1 of morin group was significantly increased after 24 hours of intervention(P＜0.05,P＜0.001).The inhibition rates of A549 cells in 30,60,90,120 and 150 μg·mL-1 of morin groups were significantly increased after 48 and 72 hours of intervention(P＜0.001).The number of A549 cell colonies and the number of green fluorescent proliferation positive cells in the 30,90,150 μg·mL-1 of morin groups were significantly decreased(P＜0.01,P＜0.001),the apoptosis rate was significantly increased(P＜0.01,P＜0.001),and the protein expression level of cleaved-PARP was significantly increased(P＜0.001).The protein expression levels of p-P38/P38 MAPK in A549 cells of 90 and 150 μg·mL-1 of morin groups were significantly increased(P＜0.01,P＜0.001).Different degrees of orange fluorescence appeared in A549 cells of 30,90 and 150 μg·mL-1 of morin groups,and the orange fluorescence of 90 and 150 μg·mL-1 of morin groups was significant.Autophagosomes and autolysosomes appeared in the cytoplasm of A549 cells in 150 μg·mL-1 of morin group,respectively.The protein expression of LC3-Ⅱ in A549 cells of 150 μg·mL-1 of morin group was significantly up-regulated(P＜0.05).The protein expression of Atg16L1-Ⅱ in A549 cells of 90,150 μg·mL-1 of morin group was significantly up-regulated(P＜0.001),and the protein expressions of p-mTOR/mTOR and p-STAT3/STAT3 were significantly down-regulated(P＜0.001).Compared with the morin(150 μg·mL-1)group,the survival rate of A549 cells in the morin(150 μg·mL-1)+chloroquine(10 μg·mL-1)group was significantly increased(P＜0.05).Conclusion Morin can promote the apoptosis of A549 cells and induce autophagy in A549 cells,and the mechanism may be related to mTOR/STAT3 axis.

Objective To explore the risk factors for intracardiac thrombosis in dilated cardiomyopathy(DCM)pa-tients and to construct,validate,and evaluate a nomogram prediction model based on these factors.Methods 88 patients diagnosed with DCM and complicated with intracardiac thrombus,and 544 patients without intracardiac thrombus were included.The participants were randomly divided into training and validation sets at a ratio of 7∶3.Using both univariate and multivariate Logistic regression analyses,independent risk factors for intracardiac thrombosis in DCM patients were identified.A nomogram prediction model was constructed using R software.The model's validity and performance were assessed using the receiver operating characteristic(ROC)curve,the Hos-mer-Lemeshow goodness-of-fit test,calibration curve,and decision curve.Results The binary Logistic regression analysis showed that age,atrial fibrillation,left ventricular end-diastolic diameter(LVEDD),brain natriuretic peptide(BNP),and β-blockers were independently associated with intracardiac thrombosis in DCM patients.Based on these five factors,a nomogram was constructed and validated.The area under the ROC curve for the training set was 0.823(95%CI:0.760～0.887)and0.803(95%CI:0.705～0.901)for the validation set,in-dicating a good discriminative ability.The Hosmer-Lemeshow test results for the calibration curve were(χ2=6.679,P=0.572)for the training set and(χ2=2.588,P=0.958)for the validation set,indicating a good fit between predicted and observed outcomes.The decision curve showed a high net clinical benefit in the threshold range of 0.05～0.92.Conclusion Based on age,atrial fibrillation,LVEDD,BNP,and β-blockers,the nomo-gram prediction model exhibits good discriminative and calibration abilities,and high clinical benefit.It can effec-tively guide clinicians in early intervention of risk factors,reducing the risk of intracardiac thrombosis in DCM pa-tients.

Objective: To investigate the mediating effects of resilience and emotion regulation on family cumulative risk and positive coping style among primary and middle school students, so as to provide insights into promoting psychological health among them. Methods: The primary and middle school students from three counties (districts) of Baise City, Guangxi Zhuang Autonomous Region were selected through the stratified cluster random sampling method. Resilience, emotion regulation, family cumulative risk and positive coping style were investigated using the Resilience Scale for Chinese adolescent, Emotion Regulation Questionnaire, Chinese family Assessment Instrument, and Simple Coping Strategies Questionnaire, respectively. Mediating effects of resilience and emotion regulation on family cumulative risk and positive coping style were examined using Process macro program. Results: Totally 1 076 questionnaires were allocated, and 1 006 valid questionnaires were recovered, with an effective rate of 93.49%. There were 515 boys, accounting for 51.19%. The median age was 13 (interquartile range, 1) years. The mean scores of resilience, emotion regulation and positive coping style were (3.36±0.63), (2.26±0.56) and (1.85±0.39) points, respectively. The median family cumulative risk index was 0 (interquartile range, 0). The results of mediating effects showed that family cumulative risk had a direct negative effect on positive coping style (β=-0.037, 95%CI: -0.059 to -0.014, P=0.011), and also had an indirect negative effect on positive coping style through psychological resilience (β=-0.029, 95%CI: -0.041 to -0.017, P=0.006) as a mediator or through psychological resilience and emotion regulation (β=-0.002, 95%CI: -0.004 to -0.001, P=0.001) as a chain mediator. The mediating effect of resilience contributed 44% to the total effect, while the chain mediating effect of resilience and emotion regulation contributed 3%. Conclusion Resilience and emotional regulation serve as negative mediators between family cumulative risk and positive coping style among middle and primary school students.

ObjectiveTo explore the therapeutic effect and mechanism of the Danhe granules on hypercholesterolemia rats by observing the changes in the efficacy indicators and the levels of proteins related to the cholesterol metabolism pathway in the rats under the intervention of Danhe granules. MethodSD rats were randomly assigned to either the blank group or the model group based on their body weight. The blank group had normal chow diets， while the model group was fed high-fat diets for seven weeks. One week after the establishment of the model， the content of the serum total cholesterol （TC） in the model rats was detected. According to the TC value， the model group was further randomly divided into a control group， pravastatin sodium tablet group（4.02 mg·kg^-1）， Xuezhikang capsule group（0.12 g·kg^-1）， high-dose， middle-dose， and low-dose groups of Danhe granules（4.536， 2.268， 1.134 g·kg^-1）. After grouping the model groups， each treatment group received continuous oral gavage for six weeks， with weekly measurements of body weight and food intake （the difference between feed intake and feed surplus）. Six weeks later， the levels of serum TC， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were measured. The liver pathology and lipid droplet distribution were evaluated by hematoxylin-eosin （HE） staining and oil red O staining， with scoring and calculation conducted. Rat liver tissue was collected， and western blot and immunohistochemistry （IHC） were used to detect the expression levels of cholesterol metabolism-related proteins namely phosphorylated adenosine 5'-monophosphate （AMP）-activated protein kinase （p-AMPK）， AMPK， 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGCR）， low-density lipoprotein receptor （LDLR）， cholesterol 7α-hydroxylase （CYP7A1）， Acyl-coenzyme A： cholesterol acyltransferase 2 （ACAT2）， and apolipoprotein B （ApoB） in hypercholesterolemia rats. ResultCompared with the blank group， the model group showed a significantly higher level of serum TC （P<0.01）. The TG level had no significant change， and the HDL-C level was significantly decreased （P<0.05）. The liver index， steatosis score， total score of pathological state， and the positive area ratio of oil red O staining were significantly increased （P<0.01）， and the protein expression levels of p-AMPK， p-AMPK/AMPK， LDLR， and CYP7A1 were significantly decreased （P<0.05， P<0.01）， while the protein expression levels of AMPK， HMGCR， and ACAT2 were significantly increased （P<0.05， P<0.01）. Compared with the model group， the TC level in each dose group of Danhe granules was significantly decreased （P<0.05）， and the positive area ratio of oil red O staining in the pravastatin sodium tablet group and medium-dose group of Danhe granules was significantly decreased （P<0.05）. In each administration group， the protein expression levels of p-AMPK and p-AMPK/AMPK were significantly increased （P<0.05， P<0.01）， and the levels of HMGCR and ACAT2 were significantly decreased （P<0.01）. The ApoB level showed a downward trend. The CYP7A1 level in the pravastatin sodium tablet group and each dose group of Danhe granules was significantly increased （P<0.05， P<0.01）， and the LDLR level in the pravastatin sodium tablet group， Xuezhikang capsule group， and high-dose and medium-dose groups of Danhe granules was significantly increased （P<0.05， P<0.01）. ConclusionDanhe granules can reduce serum TC levels and improve hepatic steatosis. It may activate AMPK， down-regulate the expression of HMGCR， and inhibit cholesterol synthesis. It can also up-regulate the expression of LDLR and CYP7A1， promote cholesterol uptake and excretion， down-regulate the expression of ACAT2 and ApoB， reduce cholesterol absorption and assembly of LDL and other lipoproteins， and thus play a role in the treatment of hypercholesterolemia.

Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

Objective To optimize the screening methods for antiallergic activity of two Chinese medicines in vitro and compare the antiallergic activity of five medicine pairs in vitro.Methods The degranulation assay of RBL-2H3 cells was optimized,and the activity of the five drugs on inhibiting mast cell degranulation was compared by toluidine blue staining and the release of β-HEX and histamine(HIS).The hyaluronidase inhibition test was optimized to compare the hyaluronidase inhibition effect of five Chinese medicine pairs.Results In the experiment of degranulation of RBL-2H3 cells,5 medicine pairs could inhibit β-HEX release from model cells to different degrees(P<0.05),including Schizonepetae Herba-Saposhnicovia divaricata,Ephedrae Herba-Asarum,Saposhnicovia divaricata-Radix Angelicae dahuricae,Rhizoma Chuanxiong-Asarum.The β-HEX release rate in the supernatant of degranulated model cells was significantly increased(P<0.05)after coculture with Flos Magnoliae-Fructus Xanthii.All the five medicine pairs could reduce HIS release in the supernatant of degranulated model cells.In the hyaluronase inhibition rate test,the hyaluronase inhibition rate of each medicine pair was Rhizoma Chuanxion-Asarum>Saposhnicovia divaricate-Radix Angelicae dahuricae>Schizonepetae Herba-Saposhnicovia divaricate>Ephedrae Herba-Asarum,among which Rhizoma Chuanxiong-Asarum had the strongest anti-allergic activity.Conclusion The trend of antiallergic activity of different drugs obtained by the two methods is consistent in vitro,indicating that the two methods can be used for screening antiallergic activity in vitro.

Objective:To investigate the relationship between carotid atherosclerosis(CAS)and subclinical left ventricular(LV)dysfunction in type 2 diabetes mellitus patients with preserved LV ejection fraction(LVEF).Methods:A total of 120 patients with type 2 diabetes mellitus who had LVEF≥50% were selected in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University from June 2021 to October 2021. The global longitudinal strain(GLS)was obtained by two-dimensional speckle tracking echocardiography(STE)to assess subclinical LV systolic function. The mitral ratio of peak early to late diastolic filling velocity(E/A), and mitral velocity to early diastolic velocity of the mitral annulus(E/E′)ratio were obtained by pulsed tissue Doppler echocardiography to assess LV diastolic function. Acrroding to bilateral carotid ultrasound examination, the subjects were divided into normal carotid arteries group( n=46) and CAS group( n=74). Demographics and biochemical parameters were compared between two groups. Binary logistic regression and Pearson correlation analysis were used to evaluate the relationship between CAS and subclinical LV dysfunction. Results:The CAS group had a higher proportion of men, older age, and a longer duration of diabetes than the normal carotid arteries group(all P<0.05). There was no difference in LVEF and GLS between the two groups [normal carotid arteries group vs CAS group, LVEF: (60.72±4.73)% vs(60.07±4.28)%; GLS: (18.24±3.72)% vs(17.81±3.47)%, respectively; both P>0.05]. However, compared with normal carotid arteries group, E/A ratio was decreased and E/E′ ratio was significantly increased in CAS group(both P<0.01). Pearson correlation analysis showed that GLS was not correlated with carotid plaque thickness and carotid intima-media thickness(CIMT; both P>0.05). By contrast, E/E′ ratio was positively correlated with carotid plaque thickness and CIMT(both P<0.05). Binary logistic regression analysis showed that GLS and E/E′ ratio were not associated with CAS( both P>0.05). However, decreased E/A ratio was significantly associated with the existence of CAS( OR=0.09, 95% CI 0.01-0.67, P=0.018). Conclusions:In type 2 diabetes mellitus patients without overt heart failure and with preserved LVEF, the occurrence of CAS is not associated with subclinical LV systolic impairment assessed by GLS, but is significantly associated with LV diastolic dysfunction, and is independent of traditional cardiovascular risk factors.

Objective:To explore the feasibility and clinical value of holographic image technology in intracranial tumor surgery.Methods:A total of 40 patients with intracranial tumors in Beijing Tiantan Hospital, Capital Medical University from November 2020 to March 2021 were randomly divided into the experimental group (20 cases) and the control group (20 cases) based on the random number table. Patients in the experimental group underwent craniotomy assisted by holographic technology, while patients in the control group underwent conventional craniotomy. The Karnofsky performance status scores of patients before the operation and 7 days after the operation were recorded, and the self-rating anxiety scale was applied to evaluate the anxiety of the patients' families.Results:The head CT 12 h after surgery showed no tumor cavity hyperdensity in all patients. The head magnetic resonance imaging (MRI) 72 h after surgery revealed 17 cases of total resection and 3 cases of subtotal resection in the experimental group; in the control group, there were 16 cases of total resection and 4 cases of subtotal resection. In the experimental group, 1 patient's left lower limb muscle strength was grade Ⅱ after the operation and recovered to grade Ⅴ-at discharge. In the control group, the left limb muscle strength of 2 patients was grade Ⅲ after the operation and recovered to grade Ⅴ at discharge. The patients with Karnofsky scores of 60, 70, 80, 90, 100 scores were found in 1 case, 5 cases, 8 cases, 6 cases, 0, respectively of the experimental group and 0, 4 cases, 9 cases, 6 cases, 1 case, respectively of the control group before the operation, and the difference was statistically significant ( P > 0.05); the patients with Karnofsky scores of 50, 60, 70, 80, 90, 100 scores were found in 1 case, 0, 0, 7 cases, 9 cases,3 cases, respectively of the experimental group and 0, 3 cases, 6 cases, 4 cases, 5 cases, 2 cases, respectively of the control group after the operation, and the difference was statistically significant ( P = 0.018). The difference of Karnofsky score before and after the operation in the experimental group was statistically significant ( P = 0.029), while there was no statistically significant difference in the Karnofsky score before and after the operation in the control group ( P = 0.241). There were 8 cases, 9 cases, 2 cases and 1 case of non-anxiety, mild anxiety, moderate anxiety, severe anxiety, respectively in the experimental group and 3 cases, 4 cases, 8 cases and 4 cases of non-anxiety, mild anxiety, moderate anxiety, severe anxiety, respectively in the control group before the operation; there were 9 cases, 9 cases, 2 cases and 0 case of non-anxiety, mild anxiety, moderate anxiety, severe anxiety, respectively in the experimental group and 2 cases, 5 cases, 9 cases and 4 cases of non-anxiety, mild anxiety, moderate anxiety, severe anxiety, respectively in the control group after the operation. The preoperative and postoperative anxiety scores of the experimental group were all lower than those of the control group, and the differences were statistically significant (preoperative P = 0.016,postoperative P = 0.002). Conclusions:Holographic technology can assist in formulating an accurate surgical plan before intracranial tumor surgery, intuitively display the anatomical relationship between the tumor and its surrounding important tissues during operation, reduce the surgical side injuries and decrease the anxiety of the patients' family.

Objective:To explore the mediating roles of family resilience and perceived social support between fear of progression and quality of life in patients with adult pulmonary hypertension, so as to provide guidance for grassroots medical staff to deeply understand and improve the long-term quality of life of adult pulmonary hypertension patients.Methods:Using a convenience sampling method, 219 adult pulmonary hypertension patients who attended Qilu Hospital, Shandong University from July 2021 to February 2022 were selected for a cross-sectional study using the World Health Organization Quality of Life Questionnaire Abbreviated Version (WHOQOL-BREF), Perceived Social Support Scale (PSSS), Shortened Chinese version of the Family Resilience Assessment Scale (FRAS-C) and Chinese version of Fear of Progression Questionnaire-Short Form (FoP-Q-SF) for investigation. A structural equation model was established to evaluate the mediating effect of family resilience and perceived social support on fear of progression and quality of life.Results:The scores of WHOQOL-BREF, PSSS, FRAS-C, and FoP-Q-SF were (45.85 ± 10.22), 66(56, 75), 94(89, 97) and 39(32, 46) points in adult pulmonary hypertension patients. The total scores of WHOQOL-BREF were negatively correlated with FoP-Q-SF ( r = -0.63, P<0.01), and positively correlated with FRAS-C and PSSS ( r=0.54, 0.46, both P<0.01). Family resilience played a partial mediating role between fear of progression and quality of life, accounting for 13.43% of the total effect. Family resilience and perceived social support played a chain mediating role between fear of progression and quality of life, accounting for 2.71% of the total effect. Conclusions:Family resilience-perceived social support play mediating roles between fear of progression and quality of life, healthcare workers can alleviate fear of progression and improve quality of life by promoting the levels of family resilience and perceived social support of adult pulmonary hypertension parents.

ObjectiveTo evaluate the effects of dehydrocostus lactone （DL） on the proliferation， apoptosis， and autophagy of human lung cancer cell A549 and to elucidate its related mechanism. MethodThe effect of DL with different concentrations （0， 5， 10， 15， 20， 25 μmol·L^-1） on the proliferation of human lung cancer A549 cells was investigated by cell counting kit-8 （CCK-8）， and its impact on the clonogenic ability of A549 cells was studied by cell clonogenic assay. The concentrations 10， 20 μmol·L^-1 were selected as DL low-dose group and high-dose group. Hoechst 33258 staining and western blot were used to observe the effect of DL on apoptosis of A549 cells. Autolysosomes were detected by acridine orange staining， and the expression level of microtubule-associated protein 1 light chain 3 （LC3） was determined by immunofluorescence and western blot. In addition， the effects of DL in combination with autophagy inhibitors bafilomycin A1 （BAF-A1） or 3-methyladenine （3-MA） on the autophagy of A549 cells was checked by CCK-8 assay. Finally， the role of DL in the regulation of A549 cell signaling pathway was explored by Western blot. ResultCompared with the conditions in the control group， the survival rate of A549 cells in the DL groups （10， 15， 20， 25 μmol·L^-1） was decreased （P<0.01）， and 5 μmol·L^-1 DL could inhibited the formation of A549 clone cells （P<0.01）， indicating that DL could inhibit the proliferation of human lung cancer A549 cells. The number of apoptotic cells was higher in both DL low-dose and high-dose groups than that in the control group， and the expression of apoptosis-related proteins poly （ADP ribose） polymerase （PARP） and B lymphocytoma-2 （Bcl-2）-associated X protein （Bax） were up-regulated （P<0.05， P<0.01）， while the expression of Bcl-2 was down-regulated （P<0.01） in DL high-dose group. The acridine orange staining showed that the orange fluorescence in the DL high-dose group was enhanced compared with that in the control group， indicating that DL could dramatically promote the formation of autolysosomes. Moreover， 20 μmol·L^-1 DL could increase the orange fluorescent particles of LC3 and up-regulated the expression level of LC3 Ⅱ （P<0.01）. After addition of autophagy inhibitors， the sensitivity of A549 cells to the effects of DL was attenuated （P<0.01）， which suggested that autophagy was involved in DL-induced A549 cell death. Compared with the control group， DL high-dose group had increased expression of autophagy-related protein 3 （Atg3） and autophagy-related protein 5 （Atg5） while reduced phosphorylation levels of protein kinase B （Akt）， mammalian target of rapamycin （mTOR） and signal transducer and activator of transcription 3 （STAT3） （P<0.05， P<0.01）. ConclusionDL could activate apoptosis and autophagy to inhibit the proliferation and clonogenic ability of A549 cells via suppressing Akt/mTOR/STAT3 signaling pathway.