Glycans, as carriers of energy in the human body and important bioinformatic molecules, control the transmission of biological information through specific binding with proteins and other biomolecules. Taking advantage of this property, they have been used as targeted delivery ligands for designing and constructing glycosylation-modified targeted delivery systems, which provide new ideas for exploration and treatment in various fields such as antiviral, inhibition of pathogenic bacteria, and treatment of cancer. Compared with traditional drug therapy, glycosylation-modified targeted drug delivery systems have multiple advantages, such as high therapeutic efficiency, low drug usage, and lower toxic side effects, and have been a hot spot in the international frontiers of drug research and development in recent years. The authors summarize the current status of domestic and international research on glycosylated modified targeted delivery, and elaborate the research progress in the fields of anti-cancer drugs, antibacterial drugs, antiviral drugs, gene delivery, vaccine delivery, etc., which provides a reference to keep abreast of domestic and international research developments, and provides information support for the research and development of glycosylated modified material targeted delivery systems.

OBJECTIVE To explore the in vivo pharmacokinetics and tissue distribution of zinc in different valence forms of arsenic in young rats after arsenic exposure in young rats, thus to provide scientific data to support the risk control and protection of arsenic exposure in young children. METHODS Twenty-four 3-week-old SD rats were randomly divided into three groups: a single-dose arsenic trioxide (ATO) group, a zinc pre-administration group, and an ATO+zinc co-treatment group. The rats were treated accordingly to their group assignments. High-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP/MS) was used to measure the arsenic species and concentrations in plasma at different time points, as well as the arsenic accumulation in various organs. Pharmacokinetic parameters for different arsenic species were analyzed using DAS 2.0 software. RESULTS The pharmacokinetics of arsenic in plasma showed that 15 min after administration, the maximum concentration of trivalent arsenic (As^Ⅲ) in plasma was highest in the ATO single-dose group, followed by the zinc pre-administration group, and then the ATO+zinc co-treatment group. The zinc pre-administration group reached lower levels 2 h after administration, earlier than the ATO group. Parameters such as ρ_max, t_max, and t_1/2 indicated that zinc supplementation accelerated the oxidation of As^Ⅲ to pentavalent arsenic (As^Ⅴ). The increase in the primary methylation index (SMI), the proportion of monomethylarsonic acid (MMA%), and the proportion of dimethylarsinic acid (DMA%), along with a significant decrease in the proportion of inorganic arsenic (iAs%), suggested enhanced methylation capacity. Tissue distribution measurements showed that zinc supplementation reduced the total arsenic (TAs) concentration in the brain, lungs, kidneys, testes, liver, and spleen. Notably, the brain, lungs, kidneys, and testes showed significantly lower TAs concentrations compared with the ATO single-dose group (P<0.05). CONCLUSION Zinc can effectively reduce the bioaccumulation of arsenic metabolites in the body by increasing the metabolic rates of As^Ⅲ and As^Ⅴ in plasma and organs, enhancing the methylation efficiency to achieve the protection against arsenic toxicity.

OBJECTIVE To investigate the immunogenicity and immune-protective efficiency of influenza A mRNA vaccines based on mRNA vaccine technology platform and provide new research strategies for the development of multivalent seasonal influenza mRNA vaccines. METHODS mRNA-H1N1 encapsulated by lipid nanoparticles(LNP) was transfected into HEK 293T cells, and the expression of LNP/mRNA-H1N1 was analyzed by Western blot and immunofluorescence microscopy in vitro, respectively. Furthermore, LNP/mRNA H1N1 was administered via intramuscular injection to SPF grade BALB/c mice, and the HA specific serum IgG titers induced by LNP/mRNA H1N1 were detected by ELISA. The secretion level of cytokine IFN-γ induced by LNP/mRNA-H1N1 was detected by ELISpot, HI titers induced by LNP/mRNA-H1N1 was measured by hemagglutination inhibition (HI) assay. Finally, the immune-protective efficacy LNP/mRNA-H1N1 was detected by the homologous H1N1 virus challenge experiment. RESULTS The specific expression of antigen protein HA could be detected by Western blot and fluorescence microscope, its molecular weight was approximately 75 000. Furthermore, LNP/mRNA-NH1N1 could induce significant humoral immune response, cellular immune response, and HI titers in BALB/c mice after prime and boost immunization. More importantly, LNP/mRNA-NH1N1 could provide 100% immune protection efficiency against H1N1 virus challenge. CONCLUSION LNP/mRNA-NH1N1 is an effective influenza mRNA candidate vaccine, laying the foundation for future preclinical safety evaluation trials in large animals and providing research ideas for the development of mRNA vaccines for other subtypes of influenza or other infectious disease vaccines.

OBJECTIVE Astragaloside Ⅳ (AS-Ⅳ) is one of the main active ingredients of astragaloside, which is widely used in treating cardiovascular diseases. The zinc ion (Zn²⁺) chelating agent N, N, N', N'-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN) can induce excessive mitochondrial autophagy and lead to myocardial cell injury. This study investigated whether AS-Ⅳ could inhibit mitochondrial autophagy mediated by fun14 domain containing 1 (FUNDC1) induced by zinc deficiency, thereby reducing the damage of H9c2 cardiomyocytes, and exploring the potential intracellular signal transduction mechanism. METHODS H9c2 cardiomyocytes were cultured using routine methods, and a zinc deficiency model was induced by TPEN. The cells were then randomly assigned to the control group, TPEN group, TPEN+AS-Ⅳ group, and AS-Ⅳ group. Cell viability was assessed using the CCK8 method; cytotoxicity was measured with lactate dehydrogenase (LDH) kits. A zinc ion detection kit was utilized to quantify intracellular zinc content. The changes in mitochondrial Zn²⁺, lysosome and mitochondrial membrane potential were detected by fluorescence microscopy and fluorescence enzyme labeling. The expression levels of autophagy-related proteins microtubule-associated protein 1A/1B light chain 3 (LC3 Ⅱ/Ⅰ), FUNDC1, sequestosome-1 (P62/SQSTM1), and translocase of the outer membrane 20 (TOM20) were detected by Western blot. Immunofluorescence analysis was performed for LC3 and FUNDC1; FUNDC1 siRNA transfection was conducted. RESULTS Compared with the control group, treatment with 10 μmol·L^-1 TPEN for 4 h significantly decreased cell viability, increased LDH release, and reduced intracellular zinc content. The red fluorescence intensity of mitochondrial Rhodzin-3 and tetramethylrhodamine ethyl ester perchlorate significantly decreased, while the red fluorescence intensity of lysosomal Lyso Tracker significantly increased. The protein expressions of LC3 and FUNDC1 were markedly elevated, whereas the protein expressions of P62 and TOM20 decreased. The expression of LC3 Ⅱ/Ⅰ and FUNDC1's green fluorescent protein is significantly increased. Pretreatment with 50 μmol·L^-1 AS-Ⅳ for 2 h significantly suppressed the aforementioned processes. Silencing FUNDC1 with siRNA further enhanced AS-Ⅳ's inhibitory effect on autophagy, with statistically significant differences (P<0.05). CONCLUSION By increasing Zn²⁺ in H9c2 cells, AS-Ⅳ can inhibit TPEN-induced mitochondrial hyperautophagy mediated by FUNDC1 and inhibit the opening of mPTP, thus playing a myocardial protective role.

OBJECTIVE To develop and validate an HPLC analytical method for finerenone, based on the concept of analytical quality by design (AQbD). METHODS LC-MS was used to identify the impurity fractions of finerenone API, and then the HPLC chromatographic conditions of finerenone were screened and optimized by analytical factorial design (2^5-1), which examined several factors, such as the acetonitrile proportion at the beginning of the elution gradient, the acetonitrile proportion at the end of the elution gradient, the elution time, the column temperature and the concentration of phosphoric acid aqueous solution, to evaluate the relationship between the critical method attributes (CMAs) and the critical method parameters (CMPs), and to generate the method operable design region (MODR), and finally perform methodological validation. RESULTS The effects of each CMPs on CMAs were analyzed, and the optimized chromatographic conditions from MODR were column temperature 45 ℃, 0.06% phosphoric acid water, gradient elution starting with 5% acetonitrile and ending with 45% acetonitrile, running for 16 min. The established finerenone analysis method had good precision, stability, linearity and sample recovery results. CONCLUSION The chromatographic conditions established based on the AQbD concept can achieve a good separation of finerenone from impurities, accurately evaluate the content of finerenone, and effectively control the quality of the drug.

OBJECTIVE To develop a new strategy for the value assignment of melting point reference standards based on optimized statistical methods, in order to enhance the accuracy and reliability of the assigned values. METHODS The development process included several key steps: structural confirmation, purity analysis, homogeneity evaluation, collaborative calibration of melting point, data preprocessing, factorial analysis, difference testing, equivalence testing, and robust statistical assignment. Using JMP software for statistical analysis, the process first involved removing outliers and conducted normality tests and homogeneity of variance tests on the melting point data. Subsequently, factorial analysis was employed to identify the main factors affecting the melting point. Each factor was then individually verified through difference testing. Equivalence testing was also conducted to determine whether the differences were within an acceptable range. Finally, based on the statistical results, the method and scope of robust statistical assignment were determined to value the melting point reference standards. RESULTS The reference materials and heating rates were identified as the main factors affecting the melting point. For each of the four model drugs, the calibration results of the same reference material were excluded because these results showed significant differences from the others and were not equivalent. The results from the remaining two reference materials and the two measurement methods were combined for statistical analysis, while the results from the two different heating rates were analyzed independently. CONCLUSION The accuracy of value assignment for melting point reference standards is improved through standardized experimental operations and optimized statistical methods, and the development process is systematically organized for the first time.

OBJECTIVE To study the chemical constituents and anti-inflammatory activity of Periplocae Cortex. METHODS The chemical components in Periplocae Cortex were extracted and isolated by MCI resin column, silica gel column and gel column, and the structure of the monomeric compounds was identified by nuclear magnetic resonance spectroscopy, mass spectrometry, infrared spectroscopy, etc. The Griess method was used to test the lipopolysaccharide-induced model of inflammatory factor release from mouse monocyte macrophage RAW264.7 cells, and the anti-inflammatory activity of the compound was evaluated. RESULTS A total of 6 compounds were isolated from Periplocae Cortex, including Δ⁵-pregnene-3β,20(S)-diol-3-O-[2-O-acetyl-β-D-digital-opyranosyl-(1→4)-β-D-cymaropyranoside]-20-O-[β-D-glucopyranosyl-(1→2)-β-D-digita-lopyranoside] (1), Δ⁵-pregnene-3β,17α,20α-triol (2), periplocoside L (3), periplocoside N (4), periplocogenin (5), periplocoside M (6). Compound 1 is a new compound, named periplocoside G1. In vitro anti-inflammatory activity studies showed that 1, 5, and 6 showed antiinflammatory activities. CONCLUSION The isolated and identified compounds and their activities provide a basis for the rational development and utilization of medicinal resources.

OBJECTIVE To explore the feasibility of preparing samples for the quality control of tuberculosis interferon-gamma (IFN-γ) release assays (TB-IGRA) using in vitro expanded tuberculosis pleural fluid mononuclear cells (TB-PFMC). METHODS The cryopreserved TB-PFMC were stimulated and expanded in vitro using three expansion protocols based on CD3/CD28 antibody combined with IL-2, and the status of TB-PFMC was observed and the expansion fold was calculated. The amplification results of three different expansion protocols were compared to determine the optimal protocol. The enzyme-linked immunospot assay (ELISPOT) and enzyme-linked immunosorbent assay (ELISA)TB-IGRA kits were used to detect TB-PFMC before and after the expansion, and the changes of the results were analyzed. RESULTS Protocol 3 was determined to be optimal and used to amplify TB-PFMC from three different patients, and their expansion folds reached 3 535.00 %, 2 800.00 %, and 3 959.00%, respectively, on the 9th day. The results of two TB-IGRA kits after the expansion of TB-PFMC remained positive and no obvious non-specific reactions were observed. CONCLUSION This study proves that TB-PFMC can be successfully amplified by CD3/CD28 antibody combined with IL-2 and the optimal protocol was determined, and the expanded TB-PFMC is suitable for the preparation of reference materials and applied for the quality control of TB-IGRA products.

OBJECTIVE To develop and verify two methods for the detection of size distribution of residual Vero cell DNA in the bulk of vaccine. METHODS Based on capillary electrophoresis and microfluidic chip-based electrophoresis, a method for separation and detection of residual Vero cell DNA fragments in different size which have been purified and fluorescent-labelled was established. The developed method was verified for specificity, accuracy, repeatability, sensitivity, and used for detection of size distribution of residual Vero cell DNA fragments in the bulk of two different vaccines. RESULTS Thirteen DNA fragments of different size were separated at baseline level. The relative deviation in retention time between the control product with 200, 500, 1 000 bp DNA and the reference product were lower than 0.5%. The average relative deviations of retention time by 6 times detection of 13 DNA fragments were 0.72% and 0.11%, respectively. The LODs were 1.5 and 0.1 pg·μL^-1, respectively. The blank control showed no influence on the test. The result of the sample added with standard DNA fragments was in accordance with the theoretical expectation. No peak was detected in one kind of vaccine sample, and the other one showed two groups of peaks representing different size. CONCLUSION The developed method has good specificity and high sensitivity. It is convenient and rapid, which is suitable for the determination of size distribution of residual DNA fragments in the bulk of vaccine.

OBJECTIVE To identify Bambusae Caulis In Taenias and its adulterants using DNA barcoding,ensure the safety andefficacy of clinicalapplications and provide a referencefor the development and quality evaluation of Bambusae Caulis In Taenias-containing drugs. METHODS By using accurately original plants bamboo samples as a reference, and based on the screening of universal barcodes, variable regions or loci within the chloroplast genome were identified to design genus-specific DNA barcodes for the molecular identification of commercial Bambusae Caulis In Taenias. RESULTS ITS2 region effectively identified Phyllostachys edulis and allowed for the preliminary identification of other species at the genus level. Additionally, 17 genus-specific DNA barcodes were designed, enabling the precise identification of Phyllostachys nigra var. henonis, Bambusa beecheyana var. Pubescens and Bambusa textilis. Among 31 batches of commercial Bambusae Caulis In Taenias, only 1 batch was authenticated as Bambusa beecheyana var. Pubescens, while the others were all adulterants, including 19 batches were identified as Phyllostachys edulis, 8 as Bambusa textilis, and 3 as other species. Furthermore, several samples were found to be mixtures of different species. CONCLUSION The method established in this study provides an accurate tool for the identification of Bambusae Caulis In Taenias original plants, laying a foundation for quality control and market regulation.