OBJECTIVE To study the accumulation pattern of effective components in Cistanche deserticola in different months, and to provide theoretical support for the harvesting, processing and quality control of Cistanche deserticola. METHODS Fifty-four batches of Cistanche deserticola samples were collected from April to December 2023, and a high performance liquid chromatography(HPLC) method was established for the simultaneous determination of six components in Cistanche deserticola, such as echinacoside, acteoside, tubuloside A, isoacteoside, poliumoside and geniposide, etc. The content of 54 batches of herbs were determined to analyze the trend of content changes. 54 batches of Cistanche deserticola herbs were identified and classified by chemometrics analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA). RESULTS The established HPLC method was able to achieve a good separation of six components in the samples. The contents of the echinacoside, acteoside, tubuloside A, and isoacteoside, decreased gradually with the blooming of the herbs in April-August and increased in September-December while the contents of poliumoside decreased in the blooming stage of Cistanche deserticola. The content of geniposide reached its highest in the full bloom period; the content of the cyclic enol ether terpene constituent geniposide was higher in November-December than in the other periods. From the HCA and PCA analyses, it was found that 54 batches of Cistanche deserticola samples could be clearly differentiated into four groups(elongation period, full bloom period, fruit splitting period, and dormant period), among which all of them were clustered together in one group during the spring harvesting period, and the main feature was that the content of echinacoside was the highest; the samples of Cistanche deserticola samples were clustered together in one group during the autumn harvesting period, and the main feature was the lowest content of acteoside; most of the Cistanche deserticola samples at the full bloom stage were clustered into one group; all the samples at the dormant stage were clustered into one group, and the main characteristic was the highest content of geniposide.The results showed that the contents of the constituents in different growth stages of the herbs of Cistanche deserticola were significantly different. From the results of OPLS-DA analysis, it can be concluded that echinacoside and acteoside are the two components that contribute more to the different growth periods, which are the key components to distinguish the different growth periods, and can be used as the index components for evaluating the quality of Cistanche deserticola herbs, so as to further differentiate the different growth periods. CONCLUSION The HPLC method established in this study has good accuracy, and combined with chemometric analysis, it can be used to distinguish the differences in the quality of the herbs of Cistanche deserticola in different months, and provide a reference for its harvesting and quality control.

OBJECTIVE To identify the composition of glyceryl mono-and distearate (GMD) from different sources using an LC-TOF-MS method and compare the composition and proportion of glycerides in samples. METHODS A Kinetex C₁₈ column (2.1 mm×100 mm, 2.6 μm) was used as analysis column. Methanol-acetonitrile-water(1∶1∶1, containing 5 mmol·L^-1 ammonium acetate) was used as mobile phase A, and isopropanol(containing 5 mmol·L^-1 ammonium acetate) was used as mobile phase B, gradient elution was performed at a flow rate of 0.3 mL·min^-1. The analysis was carried out in ESI positive and negative ion full scanning(TIC) mode and quantified by area normalization method. RESULTS The composition and proportion of 19 fatty acid glycerides in six different sources of GMD showed significant differences in the composition and proportion of glycerides. CONCLUSION This method can be used to determine the composition and proportion of glycerides from different sources, thus contributing to the revelation of the internal mechanism affecting the quality equivalence of GMD.

OBJECTIVE To explore the value of pharmaceutical care for patients with CKD3-5D by resident clinical pharmacists. METHODS Clinical pharmacists in the department of nephrology and three students formed a resident pharmacists team to develop standardized pharmaceutical care procedures(including admission medication reconciliation, doctor's order review, DRPs recognition, pharmaceutical care, medication education, discharge medication reconciliation, etc.) to provide pharmaceutical care for patients with 3-5D chronic kidney disease(CKD). The basic information, disease diagnosis and drug reorganization information of patients admitted to the department of nephrology of our hospital in 2024 were collected. The classification system based on the modified version of the European Foundation for Pharmaceutical Care Network(PCNE) was used to analyze the drug related problems(DRPs) of patients for assessment, intervention and statistical analysis. RESULTS A total of 141 patients were included in this study, and pharmacists found 54 DRPs, with an incidence of 38.3%. Among them, 36 cases(66.67%) were related to the effectiveness of treatment. Clinical pharmacists carried out 60 interventions, and the acceptance and complete implementation rate was 79.63%. The harmfulness grades of the 54 DRPs were as follows: 22 in grade E(40.74%), 16 in grade D(29.63%), 6 in grade C(11.11%), 6 in grade B(11.11%), and 4 in grade F(7.41%). CONCLUSION DRPs in patients with CKD3-5D are relatively common and harmful to some extent. Resident clinical pharmacists can identify and timely solve DRPs in patients with CKD3-5D through standardized pharmaceutical care, and assist physicians to ensure the safety, effectiveness and economy of drug use for patients.

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 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 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 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 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.

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 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.