OBJECTIVE To express recombinant protein PBP-ApoA1 by fusion of P-selectin banding peptide (PBP) and apolipoprotein (ApoA1) by Escherichia coli, and PBP-ApoA1 was applied to further prepare a recombinant high-density lipoprotein (HDL) loading with curcumin (Cur), named PA-rHDL-Cur, for the effective treatment of atherosclerosis (AS) by targeting to activated platelets. METHODS The soluble expression of PBP-ApoA1 was achieved using a co-expression strategy with glutathione S-transferase (GST) tag. The purified PBP-ApoA1, phospholipid and cholesterol were encapsulated with Cur to prepare PA-rHDL-Cur by thin-film hydration method. The physicochemical properties of PA-rHDL-Cur were characterized by particle size analyzer and UV spectrophotometer, while the release stability was evaluated using dialysis method. Cell viability and cellular uptake efficiency of PA-rHDL-Cur were assessed in vitro. Platelet adhesion experiments were conducted to confirm the targeting ability of PA-rHDL towards activated platelets. Furthermore, the antioxidant activity, cholesterol efflux effect, and reduction in oxidized high-density lipoprotein uptake capacity of RAW264.7 macrophages treated with PA-rHDL-Cur were investigated. RESULTS The yield of PBP-ApoA1 obtained by shake flask fermentation and purification was 1.3 g·L^-1. The resulting PA-rHDL-Cur exhibited uniform particle size with an average diameter of (165.3±29.6) nm and the Zeta potential of (-2.19±1.28) mV. The biocompatibility of this drug delivey system was satisfactory. In vitro cell experiments demonstrated that PA-rHDL-Cur effectively targeted atherosclerotic lesions, releasing curcumin to reduce oxidative stress within foam cells at the lesion site, significantly enhancing the bioavailability of Cur. Additionally, the presence of ApoA1 in PA-rHDL facilitated cholesterol efflux, thereby delaying the progression of atherosclerosis. CONCLUSION This design of biomimetic recombinant high-density lipoprotein nano-drug delivery system provides a new approach and theoretical basis for the development of novel nanocarriers against atherosclerosis.

OBJECTIVE To evaluate the safety and biodistribution characteristics of mixed activated killer (MAK) immune cells derived from the blood of tumor patients in immunodeficient mice. METHODS Immunodeficient NOG mice were injected with MAK cells 9 times via the tail vein. The mice were observed continuously for 28 d after the last administration. The clinical symptoms, body weight, food intake, hematological and serum biochemical indexes, cytokine indexes and histopathological changes in mice were investigated. In the meanwhile, the distribution characteristics of MAK cells in peripheral blood and various tissues and organs were investigated. RESULTS MAK cells had no significant effect on clinical symptoms, the injection site, body weight and food intake in mice, and could increase the level of human interferon gamma in serum of mice. MAK cells could elevate lymphocytes%, monocytes%, white blood cells and decrease basophils% in peripheral blood. Mix cell aggregates were observed in most organs. Massive expansion of MAK cells was observed in most tissues of mice after administration of MAK cells for 28 d. CONCLUSION MAK cells could proliferate and induce immune responses in immunodeficient NOG mice without obvious toxic reactions. Given the above, the data of this study can provide a reference for the non-clinical safety evaluation of related products.

OBJECTIVE To explore the effects of different data preprocessing algorithms and their combinations, different band selection algorithms and different classification methods on the performance of the model by using near infrared spectroscopy to classify the genuineness of Scutellaria baicalensis Georgi in Hebei province. METHODS A total of 138 samples of Scutellaria baicalensis Georgi were collected, and the spectral acquisition of different Scutellaria baicalensis Georgi samples was carried out by using 12 500-4 000 cm^-1 band. Firstly, the single performance and combined performance of different spectral preprocessing methods are compared. Secondly, the performance of competitive adaptive reweighted sampling (CARS), uninformative variable elimination (UVE), successive projections algorithm (SPA) and principal component analysis (PCA) in infrared spectral band selection and feature extraction are compared. Finally, the performance differences of partial least squares discriminant analysis (PLS-DA), support vector (SVM), artificial neural network (ANN), random forest (RF), traditional one-dimensional convolutional neural network (CNN) and stacked autoencoder (SAE) in establishing the attribute classification model of traditional Chinese medicine were compared. RESULTS The best preprocessing algorithm is to use mean centralization (MC) and multiple scattering correction (MSC), and the overall accuracy rate can reach 92.9 %. The optimal band selection algorithm is based on the 25-dimensional variables selected by PCA, which can increase the total accuracy by 10.7 %. The best classification algorithm is a one-dimensional CNN model established after MSC processing and PCA dimensionality reduction, which can achieve 100 % accuracy of geo-authentic binary classification. CONCLUSIONS The study of genuineness classification algorithm by infrared spectroscopy provides a fast and non-destructive detection method and reliable data analysis method for the genuineness classification of Scutellaria baicalensis, and provides a new method reference for the traceability of Chinese medicinal materials.

OBJECTVIE To provide safety basis for clinical application, and evaluate the toxicity CIK cells by repeated administration of CIK cells in severe immunodeficient NPG mouse model. METHODS The NPG mice were randomized into two groups: the main experimental group and the satellite group. Each group was divided into vehicle control group, low dose group (2×10⁶ cells per mouse) and high dose group (3×10⁷ cells per mouse). The CIK cells were given every two weeks for 6 times with 28-day recovery. During the experiment, the clinical symptoms, body weight and food intake were observed in the main experimental group. Blood samples were dissected and collected at the end of the administration period and recovery period, and the hematology, serum biochemical examination, cytokine detection, organ weighing and pathological examination were performed. The blood samples of satellite animals were collected to investigate the distribution of CIK cells before the first and last administration, 3 h, 1 d, 3 d, 10 d after the administration, and after the recovery period. RESULTS After repeated administration for 6 times at low dose, no obvious toxic reaction was observed in mice. A few CIK cells were detected in the blood at 1 day after the first dose and at some time points (3 h, day 10, day 29) after the last dose. High levels of CIK cells were detected in the blood of mice after repeated administration of 6 times at high dose. The levels of IFN-γ, TNF, IL-10, IL-2 and Il-10 in serum were increased. At the same time, significant graft-versus-host disease (GvHD) reaction was observed in mice, including the changes of clinical symptoms, body weight, food intake, hematology and serum biochemistry, mixed cell aggregation and GvHD-related lesions occurred in multiple organs of animals. CONCLUSION The data from the study indicates that no significant toxic reaction was observed at the dose of 2×10⁶ cells per mouse (the clinical dosage). GvHD associated with xenotransplantation was observed at a dose of 3×10⁷ cells per mouse, and no other related toxicity was observed. These data will facilitate CIK cells to enter into clinical trials.

OBJECTIVE To establish a callus culture system for dedifferentiation of Mahonia fortunei (Lindl.) in order to screen the key genes involved in the biosynthesis of jatrorrhizine. METHODS The contents of benzylisoquinoline alkaloids in callus at different dedifferentiation stages were determined by HPLC. Transcriptome was used to analyze the gene expression profiles of callus at different dedifferentiation processes and identify the differentially expressed genes. By analyzing the relationship between the variation of alkaloid content and the differentially expressed genes, the key genes involved in the biosynthesis of jatrorrhizine were screened. RESULTS The contents of columbamine, palmatine, jatrorrhizine and berberine increased in varying degrees during the dedifferentiation of Mahonia fortunei (Lindl.) leaves, among which jatrorrhizine increased significantly. By comparing and analyzing the transcripts of the leaves of Mahonia fortunei (Lindl.) and samples from different dedifferentiated stages, differential expression genes between different dedifferentiated stage samples were screened. Furthermore, through KEGG enrichment analysis, cluster analysis, 25 differentially expressed genes involved in jatrorrhizine biosynthesis pathway were annotated, among which NCS, 7OMT, CNMT, BBE, CAS, STOX may be potential key genes. CONCLUSION During the dedifferentiation period, the contents of columbamine, palmatine, jatrorrhizine and berberine all increase, among which the contents of jatrorrhizine increases the most significantly. Callus culture can be used as a method to produce benzyl isoquinoline alkaloids such as jatrorrhizine.

Age-related macular degeneration (AMD) is a degenerative retinal disease. AMD is divided into two major forms: dry (atrophic) AMD and wet (exudative) AMD. The most common treatment for wet AMD is intravitreal injection of anti-vascular endothelial growth factor drugs. However, the treatment can only relieve but not care, and there are some patients who aren’t adapted to this treatment of administration. The application of nanotechnology offers new strategies for improving drug delivery in AMD, where polymeric nanoparticles can provide sustained drug release, can be modified to target the lesion and increase drug target site deposition, can penetrate the ocular barrier and extend drug retention times. This paper reviews the current research advances in polymeric nanoparticles-based drug delivery systems for the treatment of AMD, providing a viable reference to the treatment of AMD.

OBJECTIVE To systematically evaluate the effectiveness and safety of safflower yellow for injection (SYI) in acute ischemic stroke (AIS). METHODS Computer searches were conducted on CNKI, Wanfang Database, VIP.com, Chinese Biomedical Literature Database, PubMed, Embase, Web of Science and Cochrane Library for randomized controlled trials (RCTs) of SYI combined with conventional Western medicine treatment as experimental group (EG) and conventional Western medicine treatment as control group (CG). The Cochrane risk of bias assessment tool was used to evaluate the methodological quality of included studies. Statistical analysis was performed using R4.3.1 software. RESULTS A total of 29 RCTs were included. Meta-analysis results showed that compared with the CG group, the total effective rate of the EG group was higher (P<0.000 1), the patients' NIHSS, Barthel, IL-8, IL-6, plasma viscosity, and hematocrit indexes were improved (P<0.05); while in terms of NO and FIB indexes, there was no statistically significant difference between the two groups (P>0.05). In terms of safety, the total incidence of adverse drug reactions in the EG group was lower than that in the CG group (P=0.001 8). The results of sensitivity analysis and publication bias analysis showed that except for the fact that the incidence of adverse reactions reversed to a point had no statistically significant difference between the two groups (P=0.173 4), the results obtained in the study were basically stable, and the possibility of publication bias was small. CONCLUSION Compared with conventional treatment, SYI combined with conventional treatment is generally more effective in terms of effectiveness and does not increase the risk of adverse reactions.

OBJECTIVE To evaluate the distribution and clearance of tumor infiltrating lymphocytes (TIL) in NOG mice. METHODS Eighty-four NOG mice were used, divided into control group and test group. The mice were given a single tail vein injection. After administration blood was collected and organs were taken at different time points, flow cytometry and real-time quantitative PCR (qPCR) were used to detect the distribution of cells in the blood and different organs. RESULTS After 1-2 d of administration, the number of CD3⁺, CD3⁺CD4⁺, and CD3⁺CD8⁺cells in the blood reached their peaks. At the same time, qPCR showed that TIL was mainly distributed in tissues such as lungs, blood, and liver, with less distribution in other tissues. On day 42 after administration, gene copies of most tissues were below the lower limit of quantitation. CONCLUSION Single tail vein injection of TIL in NOG mice is mainly distributed in the lungs, blood, and liver, and its duration in vivo does not exceed 42 d.

OBJECTIVE To describe the incidence of inappropriate medication prescription and adverse drug reactions between 2020 and 2021 in our hospital, providing new strategies to reduce medication errors in pediatrics and providing reference for pharmacovigilance. METHODS A graded response system based on hospital information system (HIS), medication instructions and clinical guideline was constructed to provide a scientific basis for clinical intervention and management by pharmacists. The frequency, types and distribution of inappropriate medication prescription were collected by retrieving electronic medical record. By frequency analysis, composition ratio analysis and other methods, the safety, rationality, and necessity of pre-prescription review system were evaluated. RESULTS The most common types of errors identified included inappropriate refills, incorrect dose/frequency, inappropriate drug-drug interaction, off-label medication, and quantity prescribed. After the pre-prescription review system was constructed, the amounts of inappropriate medication prescription (level 5-7) in outpatient and inpatient department were decreased by 63.4% and 64.7%, respectively (P<0.05). By stratified analysis, the amounts of inappropriate medication prescription (level 5-7) in neonatal intensive care unit and cardiac intensive care unit were declined markedly (P<0.05). In addition, the incidence of adverse drug reaction was declined notably, especially the general adverse drug reactions and severe adverse drug reactions, which were decreased by 22.7% and 60.0%, respectively (P<0.05). One year after the pre-prescription review system put into service, a retrospective review of inappropriate prescriptions (level 6-7) demonstrated a 71.6%-79.2% successful intervention rate by the system. CONCLUSION The application of pre-prescription review system improved the quality and efficiency of prescription review, and it also provide opportunities for pharmacists to ensure safe and optimal prescribing.

OBJECTIVE To established and optimized an analytical method for spiramycin related substances for daily testing of spiramycin related substances. METHODS A Waters Xbridge shield RP₁₈ (4.6 mm×250 mm, 3.5 μm) column was used for the experiment. The chromatographic conditions were as follows: water: 0.2 mol·L^-1 dipotassium hydrogen phosphate (adjust the pH value to 9.5 using a 1 mol·L^-1 KOH solution): acetonitrile: methanol, (10∶60∶28.5∶1.5) as mobile phase A, water: 0.2 mol·L^-1 dipotassium hydrogen phosphate (pH 9.5): acetonitrile: methanol, (10∶30∶57∶3) as mobile phase B, and gradient elution was performed. Detection wavelength 232 nm. RESULTS The established method can achieve good separation between the components of spiramycin and its related substances. The mass concentration of spiramycin Ⅰ is in the range of 0.7-1 200 μg·mL^-1, and its peak area shows a good linear relationship with concentration (r=0.999 3). The limit of detection (LOD) is 0.2 μg·mL^-1, and the limit of quantification (LOQ) is 0.7 μg·mL^-1. The RSD of the repeatability and precision test is less than 2.0%, and the test solution is stable at 5 ℃ for 24 h. CONCLUSION Compared with current analytical methods, this method has strong specificity, good sensitivity and stability, and effectively shortens the testing time and improves the testing efficiency. It can be used for the daily testing of related substances of spiramycin.