OBJECTIVE To systematically evaluate the quality of the Heat-clearing and symptom-relieving formula and screen potential marker components that influence the quality of the formula. METHODS The contents of 11 components （calycosin-7- O - β -D-glucoside， ononin， hyperoside， isoquercitrin， baicalin， baicalein， cryptotanshinone， tanshinone Ⅱ A ， tanshinone Ⅰ， senkyunolide A， ferulic acid） in the Heat-clearing and symptom-relieving formula were determined by high-performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）. Using the contents of the aforementioned components as variables， cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were conducted using OriginPro 2024 software and SIMCA 14.1 software； marker components affecting the quality of the Heat-clearing and symptom-relieving formula were then screened based on the criteria of variable importance in the projection （VIP） value＞1 and P ＜0.05. The comprehensive evaluation of 20 batches of samples was carried out using the entropy weight-technique for order preference by similarity to ideal solution（TOPSIS） and grey correlation analysis （GCA） methods. RESULTS The contents of the above 11 components were 7.993-72.866， 4.542-31.228， 727.666-1 901.884， 496.846-1 293.279， 1 995.501-6 779.150， 54.500-241.280， 150.302-304.339， 79.698-189.206， 257.118-682.418， 5.498-21.687， 7.524-26.935 μg/g. CA， PCA and OPLS-DA results showed that 20 batches of samples were grouped into 2 categories. Q1， Q3， Q4， Q7-Q9， Q12， Q15， Q16 were grouped into one category， and the rest were grouped into another category； VIP values of ferulic acid， tanshinone Ⅱ A ， baicalin， cryptotanshinone， calycosin-7- O - β -D-glucoside and ononin were all greater than 1 （ P ＜0.05）. Both the entropy weight-TOPSIS and GCA methods showed that the samples ranked in the top 11 according to the euclidean distance and relative correlation degree were Q2， Q5， Q6， Q10， Q11， Q13， Q14， Q17-Q20. CONCLUSIONS The established HPLC-MS/MS method is rapid， accurate and highly sens itive. Combined with chemical pattern recognition analysis， entropy weight-TOPSIS and GCA methods， this method can be used to evaluate the quality of the Heat-clearing and symptom-relieving formula. Ferulic acid， tanshinone Ⅱ A ， baicalin， cryptotanshinone， calycosin-7- O - β -D-glucoside and ononin may be the marker components that affect the quality of this formula. The overall quality of 11 batches of the Heat-clearing and symptom-relieving formula， including Q17， is relatively superior.

Objective : To investigate the effects of microRNA-29a(miR-29a) in mediating the regulation of dihydroartemisinin(DHA) on the immune checkpoint molecule B7H3 in lung adenocarcinoma(LUAD). Methods: The expression level and prognostic significance of B7H3 in LUAD were analyzed by public database. Small interfering RNA(siRNA) was used to knock down B7H3 in LUAD cell lines A549 and HCC827, and cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were treated with gradient concentrations of DHA(0, 5, 10, 25, 50, 100 μmol/L) for 48 h, and the half maximal inhibitory concentrate(IC50) was calculated. A549 and HCC827 cells were treated with IC50concentration of DHA for 1, 2 and 3 days, and the cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were transfected with miR-29a inhibitor. After DHA treatment, the expression level of miR-29a was detected by RT-qPCR, and the expression level of B7H3 was detected by Western blot. Results : B7H3 was overexpressed in LUAD and associated with poor prognosis. After knocking down of B7H3, the proliferation ability of A549 and HCC827 cells significantly decreased(allP<0.001). DHA inhibited the proliferation of A549 and HCC827 cells in both dose-and time-dependent manners, with IC50values of 30.16 μmol/L and 7.50 μmol/L, respectively. DHA up-regulated the expression of miR-29a in A549 and HCC827 cells(P<0.001,P<0.01), and down-regulated the expression of B7H3 in both cell lines(P<0.01,P<0.001). After transfection of miR-29a inhibitor into A549 and HCC827 cells, the expression of B7H3 was up-regulated, and the down-regulation of B7H3 by DHA was partially reversed. Conclusion miR-29a mediates the molecular regulation of DHA on B7H3 in LUAD.

Objective : To establish an enzyme-linked immunosorbent assay (ELISA) for exosome B7-H3 in plas- ma , and to explore the clinical significance and diagnostic value of exosome B7-H3 in plasma from non-small cell lung cancer (NSCLC) . Methods : The plasma of 70 NSCLC patients (NSCLC group) and 36 healthy controls (HC group) were collected . Exosomes and microvesicles in plasma were separated by ultra-fast centrifuge method , and the expression levels of B7-H3 in plasma exosomes in NSCLC groups and HC groups were compared by Western blot method . In NSCLC group , the expression levels of B7-H3 in plasma exosomes and microvesicles in NSCLC group were compared . A simple and feasible ELISA method was established to detect the expression level of exosome B7 - H3 in plasma by means of polyethylene glycol (PEG) precipitation and its clinical significance was analyzed . Lo- gistic regression model was established to predict plasma-derived exosome B7-H3 as a risk factor , and receiver op- erating characteristic curve (ROC) was used to investigate the diagnostic value of exosome B7-H3 in NSCLC . Results: For exosomes and microvesicles in plasma which were extracted by ultracentrifugation , Western blot results showed that the expression level of B7-H3 in plasma exosomes of NSCLC group was higher than that of HC group (P = 0. 032) , and the expression level of B7-H3 in plasma exosomes was higher than that of microvesicles of NSCLC group (P = 0. 012) . The expression level of exosome B7-H3 in plasma extracted by PEG precipitation was also higher in NSCLC group than that in HC group (P = 0. 024) . The expression level of exosome B7-H3 in plasma of NSCLC patients was not related to gender , age , smoking or pathological type , but was related to T stage (P = 0. 002) , N stage (P < 0. 001) , M stage (P = 0. 010) and AJCC stage (P < 0. 001) . Multivariate Logistic regres- sion analysis identified exosome B7-H3 in plasma as a risk factor for NSCLC . ROC analysis showed that the sensi- tivity of exosome B7-H3 in plasma for the diagnosis of NSCLC (0. 843) was higher than that of carcinoembryonic antigen (CEA) (0. 743) , whereas the specificity (0. 722) was lower than that of CEA (0. 833) . Combined de- tection of exosome B7-H3 and CEA (AUC = 0. 928 , 95% CI:0. 877 - 0. 979) had a higher diagnostic performance for NSCLC . Conclusion B7-H3 in plasma exosomes is related to the cancer staging of NSCLC , and the combined detection of exosome B7-H3 and CEA in plasma is conducive to the laboratory diagnosis of NSCLC .

Objective: To investigate the role of murine double minute 2(MDM2) in dihydroartemisinin′s(DHA) inhibition of lung adenocarcinoma cell proliferation and migration. Methods: CCK8 assay was used to detect the inhibitory effect of gradient concentrations of DHA(0, 5, 10, 25, 50 and 100 μmol/L) and time gradients(0, 24, 48, and 72 h) on the proliferation of lung adenocarcinoma A549 and PC9 cells, and the half maximal inhibitory concentrate(IC50) were calculated respectively. Colony formation and scratch assays were used to detect the inhibitory effects of DHA on colony formation and migration of A549 and PC9 cells. Western blot was used to detect the inhibitory effects of DHA on MDM2 expression and epithelial-mesenchymal transition(EMT)-related proteins E-cadherin and N-cadherin. The promoting effects of MDM2 on proliferation, migration and EMT of lung adenocarcinoma cells were verified by small interfering RNA-mediated knockdown of MDM2(si-MDM2). The reversal effects of MDM2 overexpression on DHA′s inhibition on the proliferation and migration of A549 and PC9 cells were observed. Results: DHA inhibited the proliferation of A549 and PC9 cells in a dose⁃ and time⁃dependent manner,with IC50 values of 30. 57 and 78. 61 μmol/L , respectively. Compared with the Control group , A549 and PC9 cells had significantly decreased colony formation (both P < 0. 01) and migration (both P < 0. 01) upon treatment with DHA. Moreover, DHA significantly inhibited the protein expression levels of MDM2 and N ⁃cadherin in A549 and PC9 cells , and upregulated the expression of E ⁃cadherin protein (both P < 0. 05) . Compared with si⁃Control ,si⁃MDM2 significantly inhibited the protein levels of MDM2 and N ⁃cadherin in A549 and PC9 cells , and upregulat⁃(both P < 0. 01) of both cells. After overexpression of MDM2 in A549 and PC9 cells , the proliferation and migra⁃ tion ability were significantly enhanced (both P < 0. 05) , and the inhibitory effects of DHA were partially reversed by MDM2 overexpression (both P < 0. 05) . Conclusion DHA effectively inhibits the proliferation and migration of lung adenocarcinoma cells , and its mechanism is associated with the suppression of MDM2.

Objective To investigate the regulatory effect of artemisinin derivative dihydroartemisinin(DHA)on anti-tumor immune function of CD8+T cells induced by non-small cell lung cancer(NSCLC)cells.Methods NSCLC A549 cells were divided into DMSO control group and DHA treatment group.A549 cells were treated with DMSO and DHA at different concentrations(25,50 and 100 μmol/L),and the optimal concentration of DHA was selected to treat A549 cells for 0,24,48 and 72 h according to half maximal inhibitory concentrate(IC50).CCK-8 method and colony formation test were used to detect the effect of DHA on the proliferation and colony formation ability of A549 cells.Peripheral blood mononuclear cells(PBMCs)of healthy individuals were isolated by density gradient centrifugation.After monocytes were removed by adhesion method,A549 cells pretreated with mitomycin C were co-cultured with PBMCs at 10:1 ratio.After 2 weeks,flow cytometry was used to detect the proportion of CD8+T cells and the expression levels of perforin and granzyme B.Results Compared with the control group,the proliferation inhibition rates of A549 cells increased after treatment with 25,50 and 100 μmol/L DHA for 24 h(P<0.01).The IC50 of DHA on A549 cells was46.26 μmol/L.According to IC50 concentration analysis,the inhibi-tion rates of A549 cells treated with 50 μmol/L DHA for 0,24,48 and 72h were 1.53%,53.50%,63.84%and 69.91%,and the cells inhibition rates of A548 cells increased compared with the previous observation time point,namely 0,24 and 48 h(P<0.01).The colony formation assay showed that the colony formation number of A549 cells in DHA treated group decreased compared with the control group(P<0.01).Flow cytometry results showed that compared with the control group,the proportion of CD8+T cells induced by A549 cells in the co-culture system and the proportion of CD8+T cells expressing perforin and granzyme B were higher in DHA pretreatment group(P<0.01).Conclusion DHA inhibits the growth of NSCLC cells and promotes anti-tumor immune response of CD8+T cells induced by NSCLC cells.

Objective:To analyze the current status and deficiencies of family bed service policies in China, for references for promoting the construction of China′s home health service system.Methods:Key words such as " family bed" and " home health services" were used to search for relevant policies(from January 1, 1984 to May 31, 2023)in Peking University Treasure Database, the State Council′s policy document repository, and official websites of health administrative departments at all levels. NVivo 11.0 software was utilized for a three-level coding process to establish a policy text analysis framework and to identify deficiencies in the construction of policies.Results:A total of 63 policy documents were included, comprising 53 provincial and municipal documents, which were mainly concentrated in economically developed provinces; After three-level coding, 72 third level nodes, 21 second level nodes, and 8 first level nodes(service objects, service providers, service methods, service content, service fees, subsidy policies, hospital bed configuration, and standardized management) were obtained. Among them, the responsibilities of service providers needed to be further clarified, the technical and innovative nature of service content was still insufficient, the charging standards and medical insurance reimbursement policies needed to be improved, the support for subsidy policies was limited, and the use of intelligent devices in bed configuration needed to be strengthened.Conclusions:China′s family bed service policy focused on eight dimensions, covering a comprehensive range of content, but there were still areas that need to be refined and improved. This study suggested that relevant departments should further clarify the responsibilities of service providers, deepen the construction of service connotations, moderately increase government support, promote the intelligent construction of services, and achieve multi-party collaboration to jointly promote the sustainable development of family bed services in China.

ObjectiveTo describe the neuropsychological development screening of 0‒2 years in Tongzhou from 2017 to 2021 so as to understand the status and trend of developmental delay （DD）. MethodsAnnual report data of 21 community health service centers in Tongzhou District from 2017 to 2021 were clustered， Chi square test was used to analyze the differences in positive rate and DD rate of children aged 0‒2 years with different ages and household registration， and Chi square trend test was used to analyze the linear trend of each age group and household registration. The Gesell test results in 762 children with developmental delay were analyzed， and Chi square test was used to compare the age distribution differences in gross motor， fine motor， language and personal-social behaviors. ResultsThe DD rate of children aged 0‒2 years in 2017‒2021 was 0.43％. A decreasing trend of DD rate in the 0‒ age group was observed （χ²=14.135， P<0.001）， while an increasing trend of DD rate in the 1‒ and <3 age groups was observed （χ²=5.375， P=0.020； χ²=5.558， P=0.018）. The DD rate of children aged 0‒2 years with Beijing household registration was higher （χ²=12.504， P<0.001）. The DD rate of gross motor was the highest in the 0‒ age group （64.60%）， the DD rate of language was the highest in the <3 age group （85.97%）， and a statistically significant difference of gross motor and language was separately found in the three age groups （χ²=183.061， P<0.001； χ²=78.450， P<0.001）. ConclusionAge and Beijing household registration are the influencing factors of DD for children aged 0‒2， and 0‒ years and <3 years are the critical periods for guidance and intervention to promote the development of gross motor and language abilities.

Uncovering conserved 3D protein-ligand binding patterns on the basis of functional groups(FGs)shared by a variety of small molecules can greatly expand our knowledge of protein-ligand interactions.Despite that conserved binding patterns for a few commonly used FGs have been reported in the literature,large-scale identification and evaluation of FG-based 3D binding motifs are still lacking.Here,we propose a computational method,Automatic FG-based Three-dimensional Motif Extractor(AFTME),for automatic mapping of 3D motifs to different FGs of a specific ligand.Applying our method to 233 naturally-occurring ligands,we define 481 FG-binding motifs that are highly conserved across different ligand-binding pockets.Systematic analysis further reveals four main classes of binding motifs corresponding to distinct sets of FGs.Combinations of FG-binding motifs facilitate the binding of proteins to a wide spectrum of ligands with various binding affinities.Finally,we show that our FG-motif map can be used to nominate FGs that potentially bind to specific drug targets,thus providing useful insights and guidance for rational design of small-molecule drugs.

Papillary thyroid carcinoma is one of the most common thyroid cancer subtypes. Although papillary thyroid carcinoma can be treated effectively with excision, microwave ablation, radioactive iodine and hormone replacement therapy, these ways are inefficient in reducing its morbidity, mortality and recurrence rates. Therefore, it is very important to seek the molecular mechanism of the occurrence and development of papillary thyroid carcinoma, to provide effectively early diagnosis, accurate treatment and better long-term prognosis for papillary thyroid carcinoma patients. This paper summarizes the molecular regulatory mechanisms of non-coding RNAs and their related signaling pathways in the occurrence and development of papillary thyroid carcinoma, in order to provide evidence for continued research of biomarkers for papillary thyroid carcinoma.

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.