ObjectiveTo compare the differences between the ultrafiltration method and the dextran gel filtration method during the purification of Tfn-modified PCL micelles by using purification efficiency and micelle purity as indicators. MethodsCoumarin-6 （C6） was used as a fluorescent probe and was loaded into HOOC-PEG-PCL to form PCL micelles by the film-dispersion method. Tfn was then conjugated to the surface of PCL micelles via an amidation reaction， resulting in two types of micelles： Tfn/PCL_H and Tfn/PCL_L. The pharmaceutical properties of the two types of micelles were characterized. The micelles were then purified through ultrafiltration method and dextran gel method respectively， and the efficiency of the two methods， along with the purity of the final micelles， was compared. The density of Tfn on the surface of PCL micelles was also calculated. ResultsThe hydrated diameter of PCL micelles was approximately 73 nm， and the C6 loading efficiency was around 0.046%. The size increased to 134 nm and 158 nm for Tfn/PCL_L and Tfn/PCL_H， respectively. The micelle population was monodisperse. The purification results showed that， for the ultrafiltration method， after two and one rounds of purification， the Tfn/C6 ratio stabilized at 23.6 and 3.4 for Tfn/PCL_H and Tfn/PCL_L， respectively. For the dextran gel filtration method， the Tfn/C6 ratio reached 23.7 for the Tfn/PCL_H group after two rounds of purification. However， for the Tfn/PCL_L group， the Tfn/C6 ratio increased during four rounds of dextran gel purification， and a significant difference （P = 0.042 4） was observed between the first and last filtrations. The density of Tfn in the final micelles were calculated. For the ultrafiltration method， the Tfn density of Tfn/PCL_H and Tfn/PCL_L were 94.9% and 13.8%， respectively. For the dextran gel filtration method， the density of the two micelles were 95.6% and 14.4%， respectively. For Tfn/PCL_L group， the density results revealing a statistically significant difference （P=0.000 2）. ConclusionThe purification efficiency of the two methods is comparable. However， the purity of the final micelles shows a significant difference， with the dextran gel filtration method resulting in higher purity， particularly for the Tfn/PCL_L micelles.

ObjectiveAbnormal lipids in neurons can cause the accumulation of α-synuclein（α-syn）. This study aimed to explore the mechanism of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis extract （ASH） in treating Parkinson's disease （PD） mice using lipidomics combined with network pharmacology. MethodsMice were divided into the blank group， model group and ASH （45.5 mg·kg^-1） group. Motor ability was evaluated by pole climbing time and autonomous activity count； The oxidative stress indicators were detected by enzyme-linked immunosorbent assay （ELISA）. Lipid biomarkers in brain tissues were screened and identified by ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and metabolic pathway analysis was conducted. The key targets of ASH for PD treatment were explored using network pharmacology. The Kyoto Encyclopedia of Genes and Genomes （KEGG） database was used for pathway enrichment analysis， and the "compound-reaction-enzyme-gene" network was constructed using the MetScape plugin. The protein expression levels of glutathione S-transferase P1 （GSTP1）， glutathione S-transferase Mu 2 （GSTM2）， prostaglandin peroxide synthase 1 （PTGS1）， prostaglandin peroxide synthase 2 （PTGS2）， and prostaglandin E synthase （PTGES） were validated by Western blot. ResultsCompared with the blank group， the model group showed significantly prolonged pole climbing time and reduced autonomous activity count （P<0.01）. Compared with the model group， the ASH group demonstrated significantly faster pole climbing and increased autonomous activity count （P<0.01）. The model group exhibited significantly decreased superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） levels， and increased malondialdehyde （MDA） level in brain tissues compared with the blank group （P<0.01）. The ASH group showed increased SOD and GSH-Px levels and decreased MDA level compared with the model group （P<0.05， P<0.01）. Lipidomics analysis identified 10 differential metabolites and 8 differential metabolic pathways. Network pharmacological analysis revealed 213 intersection targets between ASH components and PD， with KEGG enrichment involving the sphingolipid signaling pathway， lipid arteriosclerosis， phosphoinositide 3-kinase/protein kinase B（PI3K/Akt） signaling pathway， mitogen-activated protein kinase（MAPK） signaling pathway， and hypoxia inducible factor-1（HIF-1） signaling pathway. Integrated lipidomics and network pharmacology analysis highlighted the central role of the arachidonic acid metabolic pathway. The Western blot results showed that ASH effectively up-regulated GSTP1， GSTM2， and PTGS1 protein expression， and down-regulated PTGS2 and PTGES protein expression. ConclusionASH can ameliorate behavioral deficits， exert antioxidant effects， regulate lipid differential metabolites and the arachidonic acid metabolic pathway， thereby exerting therapeutic effects in PD model mice.

Objective: Scutellarin in Shuganning injection has potential allergenicity and is prone to causing anaphylactoid reactions. There is an urgent need to establish a simple and rapid chromatographic quantitative method to achieve the limit control of scutellarin in Shuganning injection.
Methods: For scutellarin, HPLC method was used in the quantitative analysis using a Shim-pack GIS C₁₈ (2.1 mm×150 mm, 5 μm) column. The mobile phase was a blend of 0.1% trifluoroacetic acid in water and acetonitrile, and gradient elution progress was applied for the whole process. The flow rate was 0.3 mL·min^-1 and the column temperature was maintained at 37 ℃. The detecting wavelength was 320 nm.
Results: The linear regression equation was y=51 456x-27 352, the correlation coefficient r=0.999 8, namely the calibration curve was linear in the range of 1.02-102.00 μg·mL^-1; the average recovery was 103.76%; the detection and quantification limit for determining scutellarin was 0.16 μg·mL^-1 and 0.57 μg·mL^-1, respectively; the injection sample was stable at room temperature for 48 h.
Conclusion: This method is easy to operate, reproducible and has high accuracy, thus can be used for quality control of Shuganning injection.

Objective To establish an effective model for distinguishing glandular prodromal lesions(PGL)mixed with ground-glass nodules(mGGN)from minimally invasive adenocarcinoma(MIA)on CT based on artificial intelligence.Methods A retrospective analysis was conducted on the clinical and CT image data of 180 patients with lung adenocarcinoma confirmed by surgical pathology and with CT manifestations of mGGN in the First Affiliated Hospital of Wenzhou Medical University from January 2017 to June 2023,inclu-ding 66 patients with PGL and 114 patients with MIA.Patients were divided into the training set(n=144)and the test set(n=36)in an 8∶2 ratio using a completely random method.The quantitative parameters and radiomics features of the lesions in CT images were automatically extracted using artificial intelligence soft-ware(United Imaging Research Platform uRP).By incorporating the most obvious correlation features of omics through dimensionality reduction,five machine learning classifiers were established,including logistic regression(LR),support vector machine(SVM),Random forest(RF),Gaussian process(GP),and Decision Tree(DT).The classifier with the training set highest area under the curve(AUC)was selected as the best radiomics model,and output the result as radiomics score(Rad-score).The clinical information,CT morpho-logical characteristics and quantitative data of the two groups were included in the multivariate logistic regres-sion analysis to screen the independent influencing factors for effectively differentiating PGL and MIA,and a clinical model was established.Finally,a comprehensive prediction model was constructed based on Rad-score and clinical risk factors.The diagnostic performance of the three models was evaluated by using the AUC,sen-sitivity,specificity and accuracy of receiver operating characteristic(ROC)curve.Results Eleven radiomics features for distinguishing PGL from MIA were obtained through LASSO dimensionality reduction.Among the five machine learning classifiers,GP has the best diagnostic performance,with AUC of 0.865 in the train-ing set and 0.762 in the test set,respectively.Univariate and multivariate logistic regression analyses were used for clinical feature screening.The clinical model was constructed by using the average CT value,average long and short diameter,and solid partial long diameter of mGGN,and the AUCs of the training set and the test set were 0.870 and 0.794,respectively.The comprehensive prediction model demonstrated superior diag-nostic performance,with AUC,sensitivity,specificity,and accuracy in the training set being 0.948,81.1%,91.2%and 87.5%respectively,while 0.883,76.9%,91.3%and 86.1%respectively in the test set.Conclu-sion The comprehensive prediction model established based on the quantitative and omics feature analysis of pulmonary nodules by artificial intelligence can well distinguish mGGN mixed with PGL from MIA on CT,and can be used to guide clinical treatment decisions.

Aim To explore the differences in metabolic indicators among patients with intracranial atherosclerosis of different severity levels and their predictive value for cerebral artery stenosis and occlusion.Methods A total of 310 patients with suspected intracranial atherosclerosis who were treated in our hospital from February 2022 to February 2024 were selected,and they were divided into the normal group(n=155)and the occlusion group(n=155)based on whether cerebral artery stenosis and occlusion occurred.Patients in the occlusion group were divided into grade 1 group(n=40),grade 2 group(n=78)and grade 3 group(n=37)according to the grade of intracranial atherosclerosis.The clinical data and serum calcium and phosphorus metabolism indicator levels of patients with different grades of athero-sclerosis were compared.The generalized additive model(GAM)was used to analyze the relationship between the levels of serum calcium and phosphorus metabolism indicators and the grade of atherosclerosis.The clinical data of the occlusion group and the normal group were compared.Multivariate Logistic regression analysis was conducted to analyze the factors affecting cerebral artery stenosis and occlusion.The dose-response relationship between the levels of serum calcium and phosphorus metabolism indicators and cerebral artery stenosis and occlusion was analyzed.The differences in cerebral ar-tery stenosis and occlusion under different grades of atherosclerosis and different levels of serum calcium and phosphorus metabolism indicators were compared.The generalized linear model was used to analyze the influence of the severity of in-tracranial atherosclerosis on the association between cerebral artery stenosis and occlusion and the levels of serum calcium and phosphorus metabolism indicators.Results With the increase of atherosclerosis grading level,the levels of fasting blood glucose(FBG),high sensitivity C-reactive protein(hs-CRP),apolipoprotein B(ApoB),total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC),blood phosphorus,calcium-phosphorus product,and intact parathyroid hormone(iPTH)gradually increased,while the levels of apolipoprotein A(ApoA)and high density lip-oprotein cholesterol(HDLC)gradually decreased(P＜0.05).The results of GAM analysis showed that blood phosphor-us,calcium-phosphorus product and iPTH had a positive effect on atherosclerosis grading.Compared with normal group,the levels of FBG,hs-CRP,ApoB,TC,TG,LDLC,blood phosphorus,calcium-phosphorus product and iPTH were signif-icantly higher in occlusion group,and the levels of ApoA and HDLC were significantly lower(P＜0.05).Compared with the normal group,there were significantly fewer patients with atherosclerosis grade 0 in the occlusion group(P＜0.05).ApoA≤1.02 g/L,ApoB＞1.09 g/L,TC＞5.31 mmol/L,TG＞2.53 mmol/L,LDLC＞3.12 mmol/L,HDLC ≤ 1.26 mmol/L,blood phosphorus＞2.17 mmol/L,calcium-phosphorus product＞4.53(mmol/L)2,iPTH＞327.49 ng/L and atherosclerosis grade ≥1 were the risk factors for cerebral artery stenosis and occlusion(P＜0.05).The correlation intensity of blood phosphorus,calcium-phosphorus product,iPTH and cerebral artery stenosis and occlusion showed a non-linear dose-re-sponse relationship(P＜0.001).With the increase of atherosclerosis grading,the positive correlation between cerebral artery stenosis and blood phosphorus,calcium-phosphorus product and iPTH gradually increased.Conclusion There were significant differences in the levels of metabolic indicators among patients with intracranial atherosclerosis of different severity levels,and they had predictive value for cerebral artery stenosis and occlusion.

This article systematically summarizes the clinical experience of Professor Yang Jun,a nationally renowned traditional Chinese medicine(TCM)physician,in applying refined direct moxibustion(applying a moxibustion pen made by Chinese medical extract)to treat functional dyspepsia(FD)of the stuffiness-fullness type.Based on decades of clinical practice,Professor Yang innovatively established a moxibustion therapy system for FD,which centers on TCM syndrome differentiation and treatment.The system emphasizes the refined identification of epigastric stuffiness and fullness syndrome,particularly focusing on the relative significance of abdominal distension and poor appetite.Its therapeutic features lie in establishing the principle of"prioritizing mind regulation while holistically harmonizing body and spirit",combined with personalized moxibustion dosage control and a unique refined direct moxibustion technique.By optimizing the configuration of each step in moxibustion therapy,the system maximizes therapeutic efficacy,providing novel theoretical foundations and clinical strategies for moxibustion treatment of stuffiness-fullness type of FD.

This article introduces Professor Yang Jun's clinical experience in treating bronchial asthma using warming needle moxibustion via the governor vessel-unblocking and conception vessel-regulating method.Professor Yang posits that asthma pathogenesis-whether triggered by internal imbalances or external pathogens-ultimately stems from yin-yang disharmony leading to rebellious lung qi and impaired diffusion/descending functions.Thus,restoring dynamic yin-yang balance constitutes the core therapeutic principle.As the governor and conception vessels govern the body's yin-yang regulation,Professor Yang's decades of clinical practice substantiate that"harmonizing these vessels determines life's vitality".His protocol combines warming needle moxibustion with press needles to activate governor-conception vessel functions,achieving five therapeutic effects:(1)yin-yang harmonization,(2)qi movement regulation,(3)meridian unblocking,(4)visceral stabilization,and(5)pathogen elimination,demonstrating remarkable efficacy.

Chimeric antigen receptor-modified T(CAR-T)cell therapy,as a new type of cellular immunotherapy,has shown good clinical efficacy in the treatment of malignant hematological tumors,especially B-cell acute lympho-blastic leukemia.However,there are problems such as antigen loss and immune evasion in single-target selection,so multi-target therapy strategies are gradually gaining attention.Multi-target CAR-T can effectively avoid antigen escape caused by a single target by targeting multiple tumor-associated antigens at the same time,reduce the risk of recurrence,and is expected to improve the therapeutic effect.This paper primarily discusses the structural types of multi-target CAR-T cell therapy and its clinical trial applications in the treatment of B-cell acute lymphoblastic leu-kemia(B-ALL),aiming to provide future references for the treatment of B-ALL.

Objective:To analyze the drug resistance characteristics and influencing factors in human immunodeficiency virus (HIV)-1 treated patients in Henan Province.Methods:HIV-1 treated patients who had received anti-retroviral therapy (ART) for more than six months and had a viral load >200 copies/mL in the Zhengzhou Sixth People′s Hospital from January to December 2024 were enrolled. Plasma samples were collected. Partial pol region gene sequences and integrase gene sequences of HIV-1 were amplified by reverse transcription nested polymerase chain reaction. The REGA HIV-1 subtype analysis tool was used to determine the subtypes of HIV-1 isolates, and the HIV drug resistance database of Stanford University in the United States was used to analyze the genetic drug resistance mutations and antiviral drug susceptibility. The multivariate logistic regression analysis was used to analyze the factors related to drug resistance. Results:Among 933 HIV-1 treated patients with ART failure, 825 samples were successfully amplified, with the amplification success rate of 88.42%. The overall drug resistance rate was 70.06%(578/825), among which the drug resistance rates of nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside reverse transcriptase inhibitor (NNRTI), protease inhibitor (PI), and integrase inhibitor (INSTI) were 55.15%(455/825), 64.36%(531/825), 5.70%(47/825), and 2.31%(19/821), respectively. The most common drug resistance mutations included M184I/V (47.88%(395/825)), K103N/S (38.18%(315/825)), and K70E/G/N/Q/R/S/T/del (16.61%(137/825)). Multivariate analysis showed that the baseline CD4 + T cell count <200 cells/μL (adjusted odds ratio ( OR)=2.239, 95% confidence interval ( CI)1.011 to 4.960), an initial 2NRTI+ NNRTI-based treatment regimen (adjusted OR=44.332, 95% CI 5.191 to 378.593), initial 2NRTI+ PI/r (r means ritonavir)-based regimen (adjusted OR=14.391, 95% CI 1.304 to 158.805) and a change in the ART regimen (adjusted OR=5.941, 95% CI 2.373 to 14.878) were independent risk factors for drug resistance (all P<0.05). Conclusions:The drug resistance rate after virological failure in HIV-1 treated patients in Henan Province is relatively high, which is mainly characterized by NNRTI resistance. The baseline immune status and the choice of the initial treatment regimen are important factors affecting the occurrence of drug resistance. The treatment monitoring and drug resistance monitoring should be strengthened.

Aim To explore the differences in metabolic indicators among patients with intracranial atherosclerosis of different severity levels and their predictive value for cerebral artery stenosis and occlusion.Methods A total of 310 patients with suspected intracranial atherosclerosis who were treated in our hospital from February 2022 to February 2024 were selected,and they were divided into the normal group(n=155)and the occlusion group(n=155)based on whether cerebral artery stenosis and occlusion occurred.Patients in the occlusion group were divided into grade 1 group(n=40),grade 2 group(n=78)and grade 3 group(n=37)according to the grade of intracranial atherosclerosis.The clinical data and serum calcium and phosphorus metabolism indicator levels of patients with different grades of athero-sclerosis were compared.The generalized additive model(GAM)was used to analyze the relationship between the levels of serum calcium and phosphorus metabolism indicators and the grade of atherosclerosis.The clinical data of the occlusion group and the normal group were compared.Multivariate Logistic regression analysis was conducted to analyze the factors affecting cerebral artery stenosis and occlusion.The dose-response relationship between the levels of serum calcium and phosphorus metabolism indicators and cerebral artery stenosis and occlusion was analyzed.The differences in cerebral ar-tery stenosis and occlusion under different grades of atherosclerosis and different levels of serum calcium and phosphorus metabolism indicators were compared.The generalized linear model was used to analyze the influence of the severity of in-tracranial atherosclerosis on the association between cerebral artery stenosis and occlusion and the levels of serum calcium and phosphorus metabolism indicators.Results With the increase of atherosclerosis grading level,the levels of fasting blood glucose(FBG),high sensitivity C-reactive protein(hs-CRP),apolipoprotein B(ApoB),total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC),blood phosphorus,calcium-phosphorus product,and intact parathyroid hormone(iPTH)gradually increased,while the levels of apolipoprotein A(ApoA)and high density lip-oprotein cholesterol(HDLC)gradually decreased(P＜0.05).The results of GAM analysis showed that blood phosphor-us,calcium-phosphorus product and iPTH had a positive effect on atherosclerosis grading.Compared with normal group,the levels of FBG,hs-CRP,ApoB,TC,TG,LDLC,blood phosphorus,calcium-phosphorus product and iPTH were signif-icantly higher in occlusion group,and the levels of ApoA and HDLC were significantly lower(P＜0.05).Compared with the normal group,there were significantly fewer patients with atherosclerosis grade 0 in the occlusion group(P＜0.05).ApoA≤1.02 g/L,ApoB＞1.09 g/L,TC＞5.31 mmol/L,TG＞2.53 mmol/L,LDLC＞3.12 mmol/L,HDLC ≤ 1.26 mmol/L,blood phosphorus＞2.17 mmol/L,calcium-phosphorus product＞4.53(mmol/L)2,iPTH＞327.49 ng/L and atherosclerosis grade ≥1 were the risk factors for cerebral artery stenosis and occlusion(P＜0.05).The correlation intensity of blood phosphorus,calcium-phosphorus product,iPTH and cerebral artery stenosis and occlusion showed a non-linear dose-re-sponse relationship(P＜0.001).With the increase of atherosclerosis grading,the positive correlation between cerebral artery stenosis and blood phosphorus,calcium-phosphorus product and iPTH gradually increased.Conclusion There were significant differences in the levels of metabolic indicators among patients with intracranial atherosclerosis of different severity levels,and they had predictive value for cerebral artery stenosis and occlusion.