Objective: To understand the serum vitamin D and vitamin K 1 levels of children in the Qingdao area, so as to provide scientific grounds for appropriate vitamin supplementation. Methods: A total of 4 469 children aged 0-14 years old, who attended the children s healthcare outpatient clinics of a tertiary hospital in Qingdao, were enrolled in the current study between January 2023 and July 2024. The levels of vitamin D and vitamin K 1 were measured by liquid chromatography tandem mass spectrometry. The inter group differences were analyzed using Chi square test, Wilcoxon rank sum test, and Kruskal-Wallis H test. The correlation analysis of vitamin D and vitamin K 1 levels with age was performed using the Spearman correlation. Results: The serum vitamin D level among children was 28.72(22.67, 36.26)ng/mL. The vitamin D deficiency and insufficiency rates were 2.10% and 14.59 %, respectively. The serum 25-(OH)D 2 level was 0.29(0.14, 0.53)ng/mL, the serum 25-(OH)D 3 level was 27.99( 21.78 , 35.57)ng/mL and the serum vitamin K 1 level was 0.54(0.29, 1.04)ng/mL. The vitamin K 1 deficiency rate was 13.76%. Among different age stages, the serum vitamin D level was highest in infancy [37.45(30.39, 43.87)ng/mL] and lowest in school age children [22.39(18.00, 26.97)ng/mL]; the level of vitamin K 1 was highest in preschool children [0.79(0.41, 1.51) ng/mL] and lowest in school age children[0.45 (0.26, 0.76) ng/mL]; the serum vitamin D deficiency and insufficiency rates were highest in school age children (5.03% and 30.81%); the vitamin K 1 deficiency rate was highest in infancy (21.53%) ( H/χ 2=1 698.31, 253.70 , 137.85 , 583.79, 89.30, all P <0.05). Among different seasons, the serum vitamin D and vitamin K 1 levels were lowest in the winter [26.74(18.37, 35.86) and 0.50 (0.27, 0.94)ng/mL; H =50.71, 7.86]; the vitamin D deficiency and insufficiency rates were highest in the winter (5.41% and 24.80%; χ 2=59.93, 83.35) (all P <0.05). The serum vitamin D level had a moderate negative correlation with age ( r =-0.62), and there was a low positive correlation between the serum vitamin D and vitamin K 1 levels in infancy and early childhood ( r =0.21, 0.26) (all P <0.05). Conclusions The serum vitamin D and vitamin K 1 levels are lowest in school age children and in the winter, and the serum vitamin K 1 deficiency rate is highest in infancy. There is a need to focus on critical periods of infancy and school age, and strengthen interventions during the high risk winter season. The nutritional status of vitamin D and vitamin K 1 in children should be enhanced.

Structural optimization of lead compounds is a crucial step in drug discovery. One optimization strategy is to modify the molecular structure of a scaffold to improve both its biological activities and absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties. One of the deep molecular generative model approaches preserves the scaffold while generating drug-like molecules, thereby accelerating the molecular optimization process. Deep molecular diffusion generative models simulate a gradual process that creates novel, chemically feasible molecules from noise. However, the existing models lack direct interatomic constraint features and struggle with capturing long-range dependencies in macromolecules, leading to challenges in modifying the scaffold-based molecular structures, and creates limitations in the stability and diversity of the generated molecules. To address these challenges, we propose a deep molecular diffusion generative model, the three-dimensional (3D) equivariant diffusion-driven molecular generation (3D-EDiffMG) model. The dual strong and weak atomic interaction force-based long-range dependency capturing equivariant encoder (dual-SWLEE) is introduced to encode both the bonding and non-bonding information based on strong and weak atomic interactions. Additionally, a gate multilayer perceptron (gMLP) block with tiny attention is incorporated to explicitly model complex long-sequence feature interactions and long-range dependencies. The experimental results show that 3D-EDiffMG effectively generates unique, novel, stable, and diverse drug-like molecules, highlighting its potential for lead optimization and accelerating drug discovery.

Objective To analyze the risk factors for gestational diabetes mellitus(GDM)in late pregnancy and explore the correlation between inflammatory stress status and pregnancy outcomes in GDM pregnant women.Methods A total of 87 pregnant women with GDM in late pregnancy were se-lected as GDM group,and another 87 pregnant women without GDM during the same period were se-lected as non-GDM group.Univariate analysis was conducted to compare the clinical data between the GDM group and the non-GDM group,and binary Logistic regression analysis was used to explore the risk factors for GDM.Spearman correlation analysis was employed to investigate the correlation between inflammatory stress status and pregnancy outcomes in GDM pregnant women.Results Univariate a-nalysis showed that compared with the non-GDM group,the GDM group had a higher proportion of women who were older,had a weight gain of ≥15 kg during pregnancy,a history of adverse pregnancy outcomes,a family history of diabetes,gestational hypertension,anemia during pregnancy,vaginal candidiasis infection,a preference for sweet foods,and a lower proportion of women engaging in daily exercise for≥1 h(P＜0.05).Binary Logistic regression analysis revealed that older age,weight gain of≥15 kg during pregnancy,history of adverse pregnancy outcomes,family history of diabetes,gesta-tional hypertension,anemia during pregnancy and preference for sweet foods were independent risk factors for GDM in late pregnancy(P＜0.05),while daily exercise for ≥1 h was an independent protective factor(P＜0.05).Compared with pregnant women with adverse pregnancy outcomes,women with favorable pregnancy outcomes had higher levels of C-reactive protein(CRP),lymphocytes(LYM),neutrophils(NEUT),procalcitonin(PCT)and white blood cells(WBC)(P＜0.05).Spearman correlation analysis showed that the levels of inflammatory stress indicators such as CRP,LYM,NEUT,PCT and WBC were positively correlated with the risk of adverse pregnancy outcomes in GDM pregnant women(P＜0.05).Conclusion The incidence of GDM in late pregnancy is asso-ciated with maternal age,weight gain during pregnancy,history of adverse pregnancy outcomes,family history of diabetes,gestational hypertension,anemia during pregnancy,preference for sweet foods and daily exercise time.Moreover,the inflammatory stress status of GDM patients in late preg-nancy is closely related to adverse pregnancy outcomes.

Structural optimization of lead compounds is a crucial step in drug discovery.One optimization strategy is to modify the molecular structure of a scaffold to improve both its biological activities and absorption,distribution,metabolism,excretion,and toxicity(ADMET)properties.One of the deep molecular generative model approaches preserves the scaffold while generating drug-like molecules,thereby accelerating the molecular optimization process.Deep molecular diffusion generative models simulate a gradual process that creates novel,chemically feasible molecules from noise.However,the existing models lack direct interatomic constraint features and struggle with capturing long-range dependencies in macromolecules,leading to challenges in modifying the scaffold-based molecular structures,and creates limitations in the stability and diversity of the generated molecules.To address these challenges,we propose a deep molecular diffusion generative model,the three-dimensional(3D)equivariant diffusion-driven molecular generation(3D-EDiffMG)model.The dual strong and weak atomic interaction force-based long-range dependency capturing equivariant encoder(dual-SWLEE)is introduced to encode both the bonding and non-bonding information based on strong and weak atomic interactions.Addi-tionally,a gate multilayer perceptron(gMLP)block with tiny attention is incorporated to explicitly model complex long-sequence feature interactions and long-range dependencies.The experimental results show that 3D-EDiffMG effectively generates unique,novel,stable,and diverse drug-like molecules,highlighting its potential for lead optimization and accelerating drug discovery.

Objective To identify the risk factors of patients with frequent acute exacerbations of chro-nic obstructive pulmonary disease(AECOPD)and construct a prediction model based on the clinical data,provi-ding a theoretical basis for the clinical prevention and treatment.Methods A total of 25 638 COPD patients ad-mitted to the Department of Respiratory and Critical Care Medicine,the Third People's Hospital of Chengdu from January 1,2013 to May 1,2023 were selected.Among them,11 315 patients were included according to the inclusion and exclusion criteria,and their clinical characteristics were analyzed.Multivariate Logistic regression was carried out to identify the risk factors for frequent AECOPD.A nomogram model was utilized to quantify the risk of acute exacerbation,and the performance of the prediction model was assessed based on the area under the receiver operating characteristic(ROC)curve.Results In the patients with frequent AECOPD,male percentage(P＜0.001),age(P＜0.001),urban residence(P＜0.001),smoking(P＜0.001),length of stay(P＜0.001),total cost(P＜0.001),antibiotic cost(P＜0.001),diabetes(P=0.003),respiratory failure(P＜0.001),heart disease(P＜0.001),application of systemic glucocorticoids(P＜0.001),white blood cell count(P＜0.001),neutrophil percentage(P＜0.001),C-reactive protein(P＜0.001),total cholesterol(P＜0.001),and brain natriuretic peptide(BNP)(P＜0.001)were all higher than those in the patients with infre-quent AECOPD.Multivariate Logistic regression analysis revealed that age,urban residence,smoking,diabe-tes,heart disease,Pseudomonas aeruginosa infection,application of systemic glucocorticoids,antibiotics,re-spiratory failure,and elevated white blood cell count,total cholesterol,and BNP were independent risk factors for hospitalization due to frequent AECOPD.A nomogram model of hospitalization due to frequent AECOPD was constructed according to risk factors.The ROC curve was established to evaluate the performance of the model,which showed the area under the ROC curve of 0.899(95％CI=0.892-0.905),the sensitivity of 85.30％,and the specificity of 79.80％.Conclusions Frequent AECOPD is associated with smoking,heart disease,ap-plication of systemic glucocorticoids,Pseudomonas aeruginosa infection,age,low body mass index,and elevat-ed BNP.Predicting the risks of hospitalization due to frequent AECOPD by the established model can provide the-oretical support for the treatment and risk factor management of the patients.

Objective To study the inhibitory effect of Huidu Yinhua powder from the Orthodox Manual of External Medicine on methicillin-resistant Staphylococcus aureus(MRSA),virulence factor α-hemolysin(Hla)activity,and biofilm formation,and to explore the optimal ratios of Huidu Yinhua powder and provide experimental support for its use.Methods The inhibitory effects of Huidu Yinhua powder and the herbs in the formula on USA300 were analyzed by the minimum inhibitory concentration(MIC),minimum bactericidal concentration(MBC),and disk diffusion assay(K-B method).Hemolysis,neutralization,oligomerization,and Western blot assays were used to verify in which form the drug inhibits the activity of virulence factor α-hemolysin(Hla).A biofilm assay was performed to evaluate the inhibitory effect of Huidu Yinhua powder on biofilm.Orthogonal experiments were performed to explore the optimal ratio of Huidu Yinhua powder.Results Huidu Yinhua powder inhibited the MRSA strain with a MIC90 of 64 mg/mL and an MBC of 256 mg/mL with antibacterial circle diameter of(7.50±0.50)mm.Huidu Yinhua powder inhibited Hla activity by inhibiting Hla secretion.The minimum effective concentration(MEC)was 16 mg/mL,and the MEC of biofilm was 8 mg/mL.In Huidu Yinhua powder,honeysuckle and astragalus only affected the hemolytic activity of MRSA and biofilm formation without inhibiting bacterial growth.The hemolytic activity and biofilm of MEC were both 32 mg/mL.Glycyrrhiza had a strong bacterial inhibitory capacity with a MIC90 of 8 mg/mL and biofilm MEC of 1 mg/mL without showing inhibitory hemolytic activity at subinhibitory concentrations.The orthogonal experiment showed that,at a ratio of honeysuckle,astragalus,and glycyrrhiza in Huidu Yinhua powder of 1∶2∶4,the MIC90 was 16 mg/mL,MEC of hemolytic activity was 8 mg/mL and that of biofilm was 4 mg/mL,both of which were the lowest among the nine groups.Conclusions Huidu Yinhua powder affects the hemolytic activity and biofilm formation of MRSA at subinhibitory concentrations with the optimal ratio of honeysuckle,astragalus,and glycyrrhiza being 1∶2∶4.

Objective:To investigate the clinical phenotype and genetic characteristics of developmental epileptic encephalopathy 18 (DEE18) caused by SZT2 gene variants. Methods:Clinical data of 2 children with SZT2 related DEE18 who visited the Department of Pediatric Neurology, Linyi People′s Hospital in March 2020 and July 2023 were collected. The whole exome sequencing (WES) and Sanger sequencing were applied to verify the child and their parents. SWISS-MODEL software was used to perform protein 3D modeling for the selected SZT2 gene variants. Results:Both of the 2 cases showed severe global developmental delay, epileptic seizures, autism, megacephaly, facial deformity, hypotonia, corpus callosum malformation, persistent cavum septum pellucidum, and slow background activity and focal discharge in video electroencephalography. Case 1 was easy to startle and thin in stature; case 2 had immune deficiency and clustered seizures. WES results showed that case 1 carried a compound heterozygous variant of c.5811G>A (p.W1937X) (paternal) and c.9269delG (p.S3090Ifs *94) (maternal), while case 2 carried a compound heterozygous variant of c.6302A>C(p.H2101P) (paternal) and c.7584dupA (p.E2529Rfs *20) (maternal), the parents of both patients with normal clinical phenotypes. The 4 mutations mentioned above were novel variations that had not yet been reported domestically or internationally. According to the American College of Medical Genetics and Genomics variant classification criteria and guidelines, the p.S3090Ifs *94 variant was interpreted as pathogenic; p.W1937X variant was interpreted as pathogenic; p.E2529Rfs *20 variant was interpreted as likely pathogenic; p.H2101P variant was interpreted as uncertain significance. 3D modeling showed that the variant of p.H2101P resulted in a significant change in the hydrogen bond around the 2 101st amino acid encoded, leading to a decrease in protein stability. The other 3 variants led to early truncation of peptide chain and obvious changes in protein structure. Conclusions:DEE18 caused by SZT2 gene mutation is mainly an autosome recessive genetic disease, and its clinical manifestations include global developmental delay, epileptic seizures, autism, craniofacial malformation, hypotonia, epileptic discharge, corpus callosum malformation, persistent cavum septum pellucidum, shock, small and thin stature, and immune deficiency. Four novel variants related to the SZT2 gene may be the genetic etiology of DEE18 patients in this study.

Objective:To investigate the clinical phenotype and genetic characteristics of infantile epileptic spasm syndrome caused by BRWD3 gene mutation. Methods:Clinical data of a child with BRWD3 related infantile epileptic spasm syndrome who was admitted to Department of Pediatric Neurology of Linyi People′s Hospital on August 2, 2019 were collected and followed up, whole exome sequencing technology and Sanger sequencing were applied to verify the child and his parents, and the pathogenicity of mutation site was analyzed. The studies till June 2023 were searched with keywords of " BRWD3" in both English and Chinese databases of China National Knowledge Infrastructure, Wanfang, Online Mendelian Inheritance in Man, and PubMed. The clinical phenotype and genetic characteristics of patients with BRWD3 related epilepsy were summarized. Results:The patient was a 4 years and 4 months old boy, with a clinical phenotype including severe global development delay, focal seizures (the onset age was 4 months), epileptic spasm (the onset age was 6 months), autism, megacephaly, high forehead as well as hypsarrhythmia. The whole exome sequencing results showed a de novo and frameshift variation c.4318_4319del(p.Q1441Efs*20)(NM_153252) in the BRWD3 gene, and the variation was interpreted as pathogenic (PVS1+PS2+PM2) according to the American College of Medical Genetics and Genomics variant classification criteria and guidelines. A total of 7 English literature articles were retrieved reporting 16 cases of BRWD3 gene related epilepsy in children (including 1 case of infantile epileptic spasm syndrome), and there has been no report in China yet. Totally there were 17 cases of BRWD3 gene related epilepsy including this case. All the cases showed X chromosome dominant inheritance, of whom 15 cases showed minor variations, including 7 missense variations, 3 frameshift variations, 3 splicing variations, 2 nonsense variations, and the remaining 2 cases showed large segment deletions. A total of 15 different variants were found. The phenotypes of the 17 patients mainly included epileptic seizures (17/17), intellectual disability (10/17), motor development disorder (7/17), speech impairment (9/17), megacephaly (8/17), facial malformation (8/17), autism (4/17) and hypotonia (4/17). The common seizure types were found to be focal seizures, occasionally epileptic spasm seizures and tonic seizures. Conclusions:BRWD3 gene variation related epilepsy is an X chromosome dominant genetic disease with a wide clinical phenotype spectrum. BRWD3 gene mutation c.4318_4319del(p.Q1441Efs *20) could cause infantile epileptic spasm syndrome, manifested as severe global developmental delay, epileptic spasm, focal seizures, autism, craniofacial malformation and hypsarrhythmia. This research enriches BRWD3 gene mutation spectrum.

Objective:To analyze the clinical phenotypes and TSC1/TSC2 gene variations in 52 children with tuberous sclerosis complex. Methods:The clinical data of 59 children with tuberous sclerosis complex hospitalized in Linyi People′s Hospital between January 2017 and October 2022 were collected. The analysis of TSC1 and TSC2 gene variations on main family members was performed, and then bioinformatics analysis followed. The positive children were divided into TSC1 gene group and TSC2 gene group, and the difference of clinical characteristics between the two groups was analyzed. Results:Among 59 children, 52 cases were detected TSC1/ TSC2 gene variations (17 cases in the TSC1 gene group and 35 cases in the TSC2 gene group). Of the 52 children, 28 (53.8%) were male, 24 were female (46.2%); 17 (32.7%) were familial cases (10 with TSC1 gene variations and 7 with TSC2 gene variations), 35 (67.3%) were sporadic cases; 46 (88.5%) had hypomelanotic macules, 13 (25.0%) had facial angiofibromas, 5 (9.6%) had shagreen patches, 49 (94.2%) had subependymal nodules/calcifications, 47 (90.4%) had cortical nodules, 2 (3.8%) had subependymal giant cell astrocytomas, 39 (75.0%) had intellectual/developmental disabilities, 49 (94.2%) had epileptic seizures, 8 (15.4%) had cardiac rhabdomyomas, 9 (17.3%) had renal angiomyolipomas, and 4 (7.7%) had retinal hamartomas. Of the 52 children, 49 variations were detected, including 4 large fragment deletion/duplication variations, and 45 point variations; 41 pathogenic variations, 7 likely pathogenic variations, and 1 variation of uncertain significance. In this study, 16 point mutations and 1 large fragment duplication mutation which had not been reported at home and abroad, and 3 high-frequency mutation sites (p.Arg692 *, p.Arg228 *, and p.Arg1200Try) were found. There was a statistically significant difference in the proportion of familial cases [10/17 vs 7/35(20%), χ2=7.838, P=0.005], median onset age of epilepsy [38.0(0.5-134.0) months vs 8.0(0.1-63.0) months, Z=3.506 , P<0.001] and the incidence of developmental retardation/intellectual impairment [8/17 vs 31/35(88.6%), χadj2=8.423, P=0.004] between the TSC1 gene and TSC2 gene groups. Conclusions:Tuberous sclerosis compiex has widespread phenotypes, can affect every body system, especially the skin and nervous system. The pathogenic gene is TSC1/ TSC2. The TSC1 gene group has more familial cases. The TSC2 gene group has an earlier onset age of epilepsy and a higher incidence of developmental retardation/intellectual impairment. In this study, 16 novel point mutations, 1 novel large fragment duplication mutation, and 3 hotspot mutations were identified, expanding the gene variation spectrum of tuberous sclerosis complex.

OBJECTIVE To summarize and analyze the profile of the implementation of pharmaceutical service by community pharmacists for patients with chronic diseases in China. METHODS Literature was searched from CNKI， Wanfang database， PubMed （Medline）， Embase， and Scopus to collect studies about community pharmacists providing pharmaceutical services for patients with chronic diseases. The ways and contents of the implementation of pharmaceutical services for chronic diseases by community pharmacists were summarized descriptively. RESULTS A total of 75 studies were included， involving 49 trial studies and 26 cross-sectional studies. The study sites were mainly located in the developed regions of China， and the types of disease involved in the studies were mainly diabetes mellitus （n＝30） and hypertension （n＝28）； most studies used the following indexes to evaluate pharmaceutical services， such as changes in disease symptoms and related indicators（n＝35）， improvement of patient compliance（n＝34）， and the occurrence of adverse drug reactions （irrational drug use） （n＝25）. The pharmaceutical service provided by community pharmacists included medication education （84.0%）， monitoring and follow-up （64.0%）， and identifying and solving medication-related problems （58.7%）. Thirty-eight studies mentioned that pharmaceutical services were achieved through teamwork， 16 of which mentioned healthcare alliances. A few studies investigated stratified healthcare systems （n＝15） and internet-based pharmaceutical services （n＝10）. CONCLUSIONS In China， pharmaceutical services provided by community pharmacies for patients with chronic diseases are still mainly confined to economically developed areas， and the scope of services is limited to a few diseases and basic pharmaceutical practices. In the future， the implementation of precise pharmaceutical services for different diseases and patients’ disease status， the establishment of medical alliances， and the development of internet-based pharmaceutical services should become the focus of pharmaceutical services.