OBJECTIVE To provide a scientific basis for the quality evaluation and clinical application of Qingwen hufei granules. METHODS Fourteen batches of Qingwen hufei granules were used as samples to establish high-performance liquid chromatography （HPLC） fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 Edition）. The chromatographic peaks were identified and the similarity was evaluated. Cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to conduct chemical pattern recognition analysis on the 14 batches of samples. Meanwhile， the contents of neochlorogenic acid （NGA）， chlorogenic acid （CHA）， cryptochlorogenic acid （CGA）， forsythoside A （FTA）， 3，5-O-dicaffeoylquinic acid （3，5-O- DA）， 4，5-O-dicaffeoylquinic acid （4，5-O-DA）， and angoroside C （AGC） in the samples were determined by HPLC. RESULTS The methodological investigation results of both the fingerprint and the content determination complied with the relevant requirements. Fourteen common peaks were indicated in the HPLC fingerprints of the 14 batches of samples， and 7 of them were identified [NGA （peak 2）， CHA （peak 3）， CGA （peak 5）， FTA （peak 11）， 3，5-O-DA （peak 12）， 4，5-O-DA （peak 13）， and AGC （peak 14）]； the similarity of each sample was greater than 0.94. The results of CA and PCA showed that the samples could be classified into 3 categories； the results of OPLS-DA indicated that peak 4 （unknown）， peak 11 （FTA）， peak 8 （unknown）， peak 9 （unknown）， and peak 1 （unknown） were the differential components. The content ranges of NGA， CHA， CGA， 3，5-O-DA， FTA， 4，5-O-DA and AGC in the 14 batches of samples were 0.210 4-0.458 7， 0.269 1-0.506 3， 0.228 1-0.461 1， 0.443 9-1.044 6， 0.066 7-0.155 7， 0.062 8-0.143 8， and 0.057 4-0.105 7 mg/g， respectively. CONCLUSIONS The HPLC fingerprint and multi-component content determination methods established in this study are efficient and reliable， and can be used for the quality evaluation of Qingwen hufei granules.

Cyclin-dependent kinase 9 (CDK9) is a member of the transcription CDK subfamily and plays a role in transcriptional regulation. Selective CDK9 degraders possess potent clinical advantages over reversible CDK9 inhibitors. Herein, we report the first ATG101-recruiting selective CDK9 degrader, AZ-9, based on the hydrophobic tag kinesin degradation technology. AZ-9 showed significant degradation effects and selectivity toward other homologous cell cycle CDKs in vitro and in vivo, which could also affect downstream related phenotypes. Mechanism research revealed that AZ-9 recruits ATG101 to initiate the autophagy-lysosome pathway, and forms autophagosomes through the recruitment of LC3, which then fuses with lysosomes to degrade CDK9 and the partner protein Cyclin T1. These dates validated the existence of non-proteasomal degradation pathway of hydrophobic driven protein degradation strategy for the first time, which might provide research ideas for chemical induction intervention on other types of pathogenic proteins.

Disrupted bone morphogenetic protein type 2 receptor (BMPR2) signaling in endothelial cells drives pulmonary arterial hypertension (PAH). However, targeted recovery of this signaling pathway by lipid nanoparticles (LNPs) has not been explored as a therapy. Here, we employed Design of Experiments to optimize the delivery efficiency of LNPs targeting pulmonary endothelial cells developed by our laboratory, resulting in a remarkable 35-fold increase in a simplified three-component formulation without helper lipids. Administration of BMPR2 mRNA LNPs effectively reversed established PAH in two experimental rat models (monocrotaline or SU5416-hypoxia) by reversing pulmonary vascular remodeling. Specifically, BMPR2 mRNA LNPs replenished the expression of BMPR2 protein and subsequently activated downstream pathways, as confirmed by elevated levels of p-SMAD1/5/9 and ID1 proteins. The relief of pulmonary arterial occlusion was demonstrated by thinned pulmonary arterial media and decreased proportion of full muscularized vessels. Alleviation of right ventricular hypertrophy was indicated by declined Fulton index, the cross-sectional area of right ventricular cardiomyocytes as well as collagen deposition. Effective recovery of right ventricular function was evidenced by increased pulmonary artery flow acceleration time/pulmonary artery flow ejection time ratio. These findings underscore the potential of restoring BMPR2 signaling through pulmonary endothelial cell-specific LNPs for treating PAH.

Objective: To explore the impact of sleep quality, experience of childhood maltreatment, and their interaction on depressive symptoms among middle school students, so as to provide the reference for early intervention of depressive symptoms among middle school students. Methods: From September to December 2023, a questionnaire survey was conducted among 1 231 students from two secondary schools in Harbin, Heilongjiang Province by a convenient sampling method. The survey included general demographic information, Childhood Trauma Questionnaire Short Form, Pittsburgh Sleep Quality Index and Short Version of Center for Epidemiological Studies Depression Scale. The Chi square test was used to analyze the differences in depressive symptom, sleep quality and childhood maltreatment among students with different demographic characteristics. Correlation analysis was conducted using Logistic regression, and interaction analysis was performed by both additive and multiplicative interaction models. Results: The detection rate of depressive symptoms among middle school students was 22.7%, and the rate for high school students (35.2%) was significantly higher than that for middle school students (17.0%) ( χ 2=50.35, P <0.01). The detection rates of depressive symptoms among middle school students with a history of childhood maltreatment and poor sleep quality were 45.8% and 44.0%, respectively. Multivariate Logistic regression analysis showed that compared to students without a history of childhood maltreatment, students with a history of childhood maltreatment had a higher risk of depressive symptoms ( OR =4.49，95% CI =3.31~ 6.09 , P <0.01);students with poor sleep quality had a higher risk of depressive symptoms than students with good sleep quality ( OR = 5.99，95% CI =4.37~8.22, P <0.01).The interaction results showed that the presence of childhood maltreatment and poor sleep quality had an additive interaction on the occurrence of depression in middle school students. Compared with students without childhood maltreatment and having good sleep quality, students with childhood maltreatment and poor sleep quality had a 22.49 times higher risk of developing depression ( OR =22.49，95% CI =14.22~35.59, P <0.01). Conclusion Depressive symptoms among middle school students are associated with childhood maltreatment and poor sleep quality, and there is an additive interaction between childhood maltreatment and poor sleep quality on the impact of depressive symptoms.

Objective To investigate the potential categories of nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity,and analyze the relationship between various latent categories and dietary motivation,so as to provide references for clinical working staff to develop relevant dietary intervention strategies.Methods A convenient sampling method was used to enroll 270 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity from the endocrine department of a tertiary general hospital in Zhenjiang City,Jiangsu province,using basic information questionnaire,Nutrition Literacy Measurement Scale for Chinese adults,and the Regulation of Eating Behaviors Scale to collect data,using latent profile analysis to extract nutrition literacy potential categories,with general data and potential categories as independent variables,dietary motivation as the dependent variable for univariate analysis and multiple linear regression analysis.Results A total of 262 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity completed the survey,and their nutrition literacy could be divided into 3 potential categories:low nutrition literacy group(16.41％),transformation dilemma group(54.96％),and middle and high nutrition literacy group(28.63％).The results of multiple linear regression analysis showed that residency style,potential categories of nutrition literacy and presence of diabetic complications were the influencing factors of dietary motivation in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity(P＜0.05).Conclusion There was a group heterogeneity in nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity.Clinical working staff should provide targeted interventions based on the patient's potential categories of nutrition literacy to stimulate ongoing motivation to maintain a healthy diet.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

An 85-year-old male patient received immunotherapy with toripalimab (240 mg by intravenous infusion on day 1, 21 days as one treatment cycle). After one week of the first medication, the patient experienced fatigue, poor appetite, weight loss, gradually developed mild ptosis of the right eyelid, shortness of breath and palpitations after activity, which were progressively aggravated. The patient also developed unclear speech, choking on water, and difficulty breathing. Laboratory tests showed alanine aminotransferase (ALT) 93 U/L, aspartate aminotransferase (AST) 171 U/L, creatine kinase (CK) 2 982 U/L, creatine kinase isoenzyme (CK-MB) 110 U/L, lactate dehydrogenase (LDH) 603 U/L, and high-sensitivity troponin T (hs-cTnT) 243 ng/L. All indicators of thyroid function were abnormal. Based on results of the laboratory tests, neurophysiological examination, and electrocardiogram examination, combined with the patient′s clinical symptoms, immune-mediated hepatitis of grade 2, immune-mediated myocarditis of grade 3, myasthenia gravis of grade 3, immune-mediated myositis of grade 2, and thyroid dysfunction of grade 1 were diagnosed, which was considered to be induced by toripalimab. Toripalimab was stopped. After treatments with glucocorticoids, liver-protective drugs, and symptomatic treatments, above-mentioned indicators showed a downward trend. After 27 days of treatments, the patient′s clinical symptoms were improved significantly. Laboratory tests showed ALT 123 U/L, AST 68 U/L, CK 116 U/L, CK-MB 42 U/L, LDH 305 U/L. The thyroid function indicators were all normal. After 57 days of treatments, above symptoms in the patient basically disappeared, and laboratory indicators tended to be normal.

An 85-year-old male patient received immunotherapy with toripalimab (240 mg by intravenous infusion on day 1, 21 days as one treatment cycle). After one week of the first medication, the patient experienced fatigue, poor appetite, weight loss, gradually developed mild ptosis of the right eyelid, shortness of breath and palpitations after activity, which were progressively aggravated. The patient also developed unclear speech, choking on water, and difficulty breathing. Laboratory tests showed alanine aminotransferase (ALT) 93 U/L, aspartate aminotransferase (AST) 171 U/L, creatine kinase (CK) 2 982 U/L, creatine kinase isoenzyme (CK-MB) 110 U/L, lactate dehydrogenase (LDH) 603 U/L, and high-sensitivity troponin T (hs-cTnT) 243 ng/L. All indicators of thyroid function were abnormal. Based on results of the laboratory tests, neurophysiological examination, and electrocardiogram examination, combined with the patient′s clinical symptoms, immune-mediated hepatitis of grade 2, immune-mediated myocarditis of grade 3, myasthenia gravis of grade 3, immune-mediated myositis of grade 2, and thyroid dysfunction of grade 1 were diagnosed, which was considered to be induced by toripalimab. Toripalimab was stopped. After treatments with glucocorticoids, liver-protective drugs, and symptomatic treatments, above-mentioned indicators showed a downward trend. After 27 days of treatments, the patient′s clinical symptoms were improved significantly. Laboratory tests showed ALT 123 U/L, AST 68 U/L, CK 116 U/L, CK-MB 42 U/L, LDH 305 U/L. The thyroid function indicators were all normal. After 57 days of treatments, above symptoms in the patient basically disappeared, and laboratory indicators tended to be normal.

Objective To investigate the potential categories of nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity,and analyze the relationship between various latent categories and dietary motivation,so as to provide references for clinical working staff to develop relevant dietary intervention strategies.Methods A convenient sampling method was used to enroll 270 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity from the endocrine department of a tertiary general hospital in Zhenjiang City,Jiangsu province,using basic information questionnaire,Nutrition Literacy Measurement Scale for Chinese adults,and the Regulation of Eating Behaviors Scale to collect data,using latent profile analysis to extract nutrition literacy potential categories,with general data and potential categories as independent variables,dietary motivation as the dependent variable for univariate analysis and multiple linear regression analysis.Results A total of 262 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity completed the survey,and their nutrition literacy could be divided into 3 potential categories:low nutrition literacy group(16.41％),transformation dilemma group(54.96％),and middle and high nutrition literacy group(28.63％).The results of multiple linear regression analysis showed that residency style,potential categories of nutrition literacy and presence of diabetic complications were the influencing factors of dietary motivation in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity(P＜0.05).Conclusion There was a group heterogeneity in nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity.Clinical working staff should provide targeted interventions based on the patient's potential categories of nutrition literacy to stimulate ongoing motivation to maintain a healthy diet.

Objective:To evaluate the predictive value of serum 25-hydroxyvitamin D (25(OH)D) level for the clinical response and imaging response to anal fistula in patients with perianal fistulizing Crohn′s disease (PFCD) treated with ustekinumab (UST).Methods:From October 1, 2021 to June 30, 2023, 80 patients with active PFCD who received UST treatment at the Second Affiliated Hospital of Wenzhou Medical University were retrospectively collected. Harvey-Bradshaw index (HBI) was applied to evaluate the clinical activity of PFCD patients. Perianal disease activity index (PDAI) were used to evaluate the clinical outcomes of anal fistula and pelvic magnetic resonance imaging (MRI) were used to evaluate the imaging outcomes of anal fistula. Serum 25(OH)D levels were examined at week 0, 8, 16, and 24 after UST treatment. Binary logistic regression models were performed to analyze the relationship between the baseline serum 25(OH)D level and the clinical pathological characteristics. And the correlation between the serum 25(OH)D level and the clinical response to anal fistula at week 8 after UST treatment was analyzed. The relationship between clinical response and imaging response to anal fistula at week 24 was also analyzed. R software was employed to draw nomograms and calculate the C-index. Independent sample t test and chi-square test were used for statistical comparison. Results:Multifactorial binary logistic regression analysis showed that the baseline level of serum 25(OH)D was independently correlated with the baseline HBI and baseline PDAI in PFCD patients ( OR=1.45, 95% confidence interval (95% CI) 1.08 to 1.95, P=0.014; OR=1.39, 95% CI 1.01 to 1.92, P=0.042). At week 8 after UST treatment, the serum 25(OH)D level of patients with clinical response to fistula was higher than that of patients without clinical response ((21.77±6.17) μg/L vs. (16.72±6.39) μg/L), while the baseline PDAI was lower than that of patients without response (6.88±2.15 vs. 8.06±2.14), and the proportions of patients with previous failure of biologic therapy and with complex anal fistula were also lower than those of patients without response (42.4%, 14/33 vs. 66.0%, 31/47; 57.6%, 19/33 vs. 78.7%, 37/47), and the differences were statistically significant ( t=3.53 and 2.43, χ2=4.36 and 4.13; P=0.002, 0.022, 0.039 and 0.045). At week 24 after UST treatment, the serum level of 25(OH)D in patients with imaging response was higher than that in patients without response ((22.48±5.81) μg/L vs. (16.66±6.34) μg/L), and the proportion of patients with previous failure of biologic therapy and the proportion of patients with complex anal fistula was lower than that in patients without response (40.0%, 20/50 vs. 12/15; 60.0%, 30/50 vs. 14/15), and all the differences were statistically significant ( t=3.33, χ2=7.39 and 5.86; P=0.004, 0.011 and 0.038). Multifactorial binary logistic regression model analysis showed that the average serum 25(OH)D level and previous failure of biological therapy were 2 independent factors of clinical response to anal fistula at week 8 after UST treatment ( OR=1.11, 95% CI 1.02 to 1.21, P=0.012; OR=0.34, 95% CI 0.12 to 0.97, P=0.043), which were also 2 independent factors of clinical response to anal fistula ( OR=1.14, 95% CI 1.05 to 1.24, P=0.002; OR=0.30, 95% CI 0.11 to 0.89, P=0.029) and imaging response to anal fistula ( OR=1.20, 95% CI 1.05 to 1.36, P=0.006; OR=0.11, 95% CI 0.02 to 0.58, P=0.009) at week 24 after UST treatment. The nomograms showed the C-indexes of the clinical response to anal fistula at week 8 and week 24 after UST treatment were 0.78 (95% CI 0.68 to 0.89) and 0.76 (95% CI 0.64 to 0.87), respectively. The C-index of imaging response at week 24 after UST treatment was 0.85 (95% CI 0.76 to 0.95). Conclusions:In PFCD patients treated with UST, serum 25(OH)D levels and previous failure of biological therapy may independently affect the clinical response to anal fistula at week 8 and 24 after UST treatment, as well as the imaging response to anal fistula at week 24 after UST treatment.