OBJECTIVE To ensure the scientificalness and rationality of proficiency testing and measurement audit projects in the field of authenticity identification of Chinese material medica and decoction pieces, and to provide new ideas for high quality development in this field. METHODS The key technical points were analyzed in the proficiency testing and measurement audit projects of authenticity identification of Chinese material medica and decoction pieces. RESULTS Proficiency testing and measurement audit projects of authenticity identification of Chinese material medica and decoction pieces are insufficient, which cannot meet the needs of market supervision. CONCLUSION Proficiency testing providers should seriously summarize and think about the key technical points in the proficiency testing and measurement audit projects of authenticity identification of Chinese material medica and decoction pieces. By improving capabilities and service, actively organizing proficiency testing projects, and establishing measurement audit sample bank, proficiency testing providers can constantly improve the quality of the proficiency testing and measurement audit projects in the field of authenticity identification of Chinese material medica and decoction pieces, and the shortcomings of external quality control in this field will be complemented which can promote the high quality development of traditional Chinese medicine(TCM) and TCM industry.

OBJECTIVE To investigate adverse event signals associated with ofatumumab and rituximab in the treatment of multiple sclerosis(MS), provide more evidences for the clinical safe medication based on the U.S. food and drug administration's adverse event reporting system(FAERS) data. METHODS Postmarking adverse event reports of ofatumumab and rituximab were extracted to form an analysis dataset. Disproportionality analysis combined with bayesian confidence propagation neural nework (BCPNN) was conducted for adverse event signal monitoring. Adverse events meeting threshold conditions were selected and summarized for analysis. RESULTS The ofatumumab group collected a total of 72 310 adverse event reports, of which 325 showed positive signals, covering 24 system organ classes (SOCs), involving “infections and infestations” “nervous system disorders” and “musculoskeletal and connective tissue disorders”. The rituximab group collected 23 203 adverse event reports, with 311 showing positive signals, covering 25 SOC, involving “injury, poisoning and procedural complications” “infections and infestations” and “nervous system disorders”. Except for “gastrointestinal disorders” and “eye disorders”, both drugs exhibited significant differences in each SOC. CONCLUSION There are substantial differences in the risk of adverse events between ofatumumab and rituximab in the treatment of MS. Therefore, medication monitoring is required during the treatment. Due to the limitations of real-world studies for quantitative analysis, the above conclusions need to be validated through large-scale cohort studies.

Hippophae rhamnoides L. is a high-quality medicinal and edible plant with rich nutritional values and a wide range of biological activities. It has been used to improve hyperglycemia, hyperlipidemia, liver injury and prevent cardiovascular diseases. Sea buckthorn polysaccharides (SBPs) are one of the most important bioactive components of Hippophae rhamnoides L.. The preperation methods of SBPs include hot water extraction, ultrasonic-assisted extraction, microwave-assisted extraction, and flash extraction. Different preperation methods lead to different configurations and biological activities of SBPs. Furthermore, these biological activities are related to the chemical structure of SBPs, including the relative molecular weight, monosaccharide composition, glycosidic bond and three-dimensional structure. Modern pharmacological studies have demonstrated that SBPs possess various activities, including hepatoprotective, anti-inflammatory, immunomodulatory, anti-tumor, antioxidant activity, as well as the regulation of blood sugar and lipid metabolism disorder, which will exhibit excellent development value in functional food and medicament. However, few studies on the structure-function relationship of SBPs have been reported. In this paper, the preperation methods, structure characterization, pharmacological activities and the utilization of SBPs were systematically reviewed, the structure-activity relationship and application prospect were discussed, which will provide a theoretical basis for a further research and development and utilization of SBPs.

OBJECTIVE To establish the fingerprint and content determination method of Impatientis Pritzelii Rhizoma to improve the quality control method of Tujia ethnomedicinal herb Impatientis Pritzelii Rhizoma. METHODS The fingerprint was established by UPLC with ACQUITY UPLC BEH C₁₈ column. Phlorizin content assay was performed by HPLC with Eclipse XDB-C₁₈ column. The mobile phases for both methods were composed of acetonitrile and 0.1% formic acid with different gradient elution procedures. The detection wavelength was set at 283 nm. The comparative study was carried out between Impatientis Pritzelii Rhizoma and its adulterants (such as the roots and rhizomes of Impatiens pinfanensis, I. uliginosa, I. siculifer and I. noli-tangere). RESULTS Nine main chromatographic peaks in the fingerprint could be served as the characteristic peaks of Impatientis Pritzelii Rhizoma, four of which were corresponding to catechin, epicatechin, scopolamine and phlorizin, respectively. The established fingerprint could reflect the distribution of the chemical components of Impatientis Pritzelii Rhizoma, which was distinguished from four adulterants. Phlorizin was found and determined in Impatientis Pritzelii Rhizoma for the first time. The contents in 16 batches of Impatientis Pritzelii Rhizoma ranged from 0.007% to 0.085%, while no phloridin was found in the adulterants. CONCLUSION The established methods for the fingerprint and phlorizin content determination of Impatientis Pritzelii Rhizoma are specific, accurate and effective, which could be used for its quality control.

OBJECTIVE To explore the mechanism of Atractylodes macrocephala polysaccharide (AMP) in improving chronic unpredictable mild stress (CUMS) depression in mice by integrating metabolomics technology and gut microbiota analysis. METHODS CUMS depression mouse model was established. Low, medium and high dose AMP treatment were given at 0.062 5, 0.125, 0.250 g·kg^-1, respectively, fluoxetine was given at the dosage of 0.015 g·kg^-1, control group and model group were given 0.9% saline solution, and all groups were given at 5 mL·kg^-1·d^-1 of drug. The content of 5-hydroxytryptamine(5-HT) in the brain of mice was determined by enzyme-linked immunosorbent assay (ELISA), and the CUMS depression model was tested by combining sucrose preference test (SPT) results. The 16S rDNA amplicon sequencing technology was used to analyze the gut microbiota in the feces of mice in each group. LC-MS technology was used to perform non-targeted metabolomics determination of mouse serum. RESULTS Compared with the control group, the sucrose preference rate of CUMS model mice was significantly lower, and the content of 5-HT in the brain of mice was significantly reduced (P<0.01), indicating that the CUMS depression model was successfully established. After AMP treatment, the sucrose preference rate and 5-HT content of mice in each group increased (P<0.01), the gut microbiota of CUMS mice had a regulatory effect, serum metabolites were significantly changed, and 58 metabolites were significantly adjusted. Spearman correlation analysis showed that the changes in gut microbiota were significantly associated with the changes in metabolite levels. CONCLUSION AMP exerts intervention effects on CUMS depression model mice by regulating the stability of gut microbiota, upregulating the F-B ratio, and thereby regulating metabolic pathways.

OBJECTIVE To observe the inhibitory effect of ritonavir on human T-cell leukemia virus type-1 (HTLV-1) transmission and malignant proliferation of adult T-cell leukemia (ATL) cells, and explore its molecular mechanism. METHODS The proliferation and vitality of ritonavir on various leukemic cells were evaluated by CCK-8 and colony formation assay. The effects of ritonavir on HTLV-1 virus transmission were detected by flow cytometry, dual luciferase reporter gene technique, qPCR and Western blot. The effects of ritonavir on the cell cycle and apoptosis of ATL cells were examined through flow cytometry. RESULTS Ritonavir could inhibit the proliferation of four ATL cell lines and the clonal proliferation of HTLV-1 positive cell lines. The former exhibited a significant dose-effect relationship and had a more pronounced inhibitory effect on HTLV-1 positive cell lines (P<0.05). Additionally, the administration of ritonavir immediately after co-culture of HTLV-1 positive cell lines with JETWT35 cells resulted in a significant down-regulation of red fluorescent protein expression in JETWT35 and inhibited the transmission of HTLV-1 virus into recipient cells (P<0.01). Upon immediate addition of ritonavir to the co-culture system of HTLV-1 positive cell lines and Jurkat cells, there was a notable inhibition of HTLV-1-related gene Tax and other genes mRNA in recipient cells (P<0.01); however, no significant effect was observed when ritonavir was added 12 h after virus transmission. Morever, ritonavir demonstrated a does-dependent inhibition of gp46 expression on the cell membrane of the HTLV-1 positive cell line ATL-T, thereby suppressing the production of HTLV-1 virus (P<0.01). Ritonavir impeded cell progression into G₁ phase and facilitated apoptosis, with the apoptosis rate of HTLV-1 positive cell lines being significantly greater than that of HTLV-1 negative cell lines (P<0.05 or P<0.01). CONCLUSION Ritonavir exerts inhibitory effects on the production and transmission of HTLV-1 virus by diminishing the activity of WT-Luc virus promoter, suppressing the expression of HTLV-1-related virus genes (Tax, HBZ, Gag, Pol, and Env). Additionally, it inhibits the expression of the HTLV-1-positive membrane surface envelope protein subunit gp46. Futhermore, ritonavir induces apoptosis in ATL cells by arresting cell cycle in the G₁ phase, thereby effectively suppressing cell proliferation.

OBJECTIVE To establish a more sensitive, simple, accurate and stable method for determining the activity of recombinant human α-galactosidase A (rhα-GAL) based on the kinetic theory of enzyme reaction and study the assay conditions for the activity assay. METHODS The optimum conditions of the assay system were as follows: 50 mmol·L^-1 p-nitrophenyl-α-D-galactopyranoside was used as the substrate, the concentration of the enzyme was 1.67 μg·mL^-1, the reaction was accurately carried out in a water bath at 37 ℃ for 15 min, then the reaction was terminated by glycine buffer (pH 10.5), and the absorbance was measured at 400 nm using a microplate reader. RESULTS The method had good specificity. Rhα-GAL showed a good linear relationship with the enzymatic reaction rate in the range of 0.83-2.51 mg·mL^-1 (r=0.999 8). The recoveries of validation solutions at 50%, 80%, 100%, 125% and 150% concentrations were in the range of 94.2%-101.8% (n=18), and the CVs of the measured results were between 2.0% and 5.5% (n=18). The CV of 12 independent tests of the same sample was 2.21% (n=12). The effects of slight changes in water bath temperature, reaction time and substrate concentration in the reaction system on the results were investigated,confirming the good robustness of the method. The reconstituted sample showed good stability when stored at 2-8 ℃ for 48 h. p-Nitrophenol showed a good linear relationship with the absorbance in the range of 0.01-0.15 mmol·L^-1 (r=0.999 7). The recoveries of p-nitrophenol solution at five concentrations were in the range of 94.9%-105.1% (n=9),and the CVs were all below 2.0% (n=9). The activity of two rhα-GAL products was determined by this method. CONCLUSION A chromogenic substrate method was established to determine the activity of rhα-GAL and validated with good sensitivity, precision and accuracy, which can be used for the activity evaluation and quality control of the product.

OBJECTIVE To investigate the protective effect and possible mechanism of Chaihu Shugan Granules on acute liver injury in mice. METHODS Chaihu Shugan Granules was administered to mice at low, medium and high dosages (crude drug dose: 11.4, 22.8, 45.6 g·kg^-1) continuously for 7 d. Two hours after the last administration, the animal model was made with 0.2% tetrachloromethane (CCl₄) solution except the control group. The serum and liver tissues were collected after 12 h. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD), malondialdehyde (MDA), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and reactive oxygen species (ROS) in liver tissues were measured by ELISA. HE staining was conducted to reveal the histopathological changes in liver. Transcriptomics was used to obtain differentially expressed mRNA in liver tissues and enrich differentially expressed pathways, while metabolomics was used to obtain changes in liver endogenous metabolites and enriches pathways of differential metabolites using KEGG database. The expression and location of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38 MARK), kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in liver tissues were measured by immunohistochemistry and Western blot. RESULTS Compared with the model group, the serum levels of ALT, AST and MDA, IL-6, TNF-α and ROS in liver tissues of mice in each group of Chaihu Shugan Granules decreased significantly (P<0.05 or P<0.01), while the level of SOD in liver tissues increased significantly (P<0.01). The degree of necrosis and inflammatory infiltration in liver cells decreased significantly. Nqo1 gene and NAD(P)H: quinone oxidoreductase 1 (NQO1) expression were up-regulated while Ccl2 gene and monocyte chemotactic protein 1 (MCP-1) expression were down-regulated. Organic acids were significantly down-regulated and carbohydrate was significantly up-regulated, the expressions of JNK, p38 MARK and Keap1 in liver tissues were significantly decreased (P<0.05 or P<0.01), while the expressions of Nrf2 and HO-1 were significantly increased (P<0.01). CONCLUSION Chaihu Shugan Granules might have protective effect on CCl₄-induced acute liver injury in mice by activating the Keap1-Nrf2/HO-1 signal pathway.

OBJECTIVE To provide suggestions for the compliance construction of complex formulation development processes, and promote the rationality and standardization of complex formulation development processes. METHODS Sorted out the characteristics and difficulties of the complex formulation development process, combined with the GMP technical requirements and verification focus, analyzed the common non-compliance situations in the complex formulation development process, and proposed compliance suggestions. RESULTS The development of complex formulations is often limited to excipients, instruments, pharmaceutical equipment, etc., with high requirements for personnel quality. Innovative production processes are often adopted or new technologies are introduced into conventional production processes, and the preparation process is complex with multiple quality control parameters, often requiring commissioned production or inspection. Some holders of complex formulations are emerging high-tech enterprises that lack experience in full lifecycle quality management. Enterprises often experience deficiencies in key personnel training, auxiliary material use and changes, equipment validation, process changes, technology transfer, deviation management, commissioned research, and data reliability. CONCLUSION Based on the characteristics of the complex formulation product development process and the key points of registration verification, the applicant needs to strengthen quality management, ensure the scientific, reasonable, and standardized development process of complex formulations, and ensure the safety and quality controllability of complex formulations.

OBJECTIVE To establish a high-performance liquid chromatography method for simultaneous determination of linezolid and its main metabolites PNU-142300 and PNU-142586 in human plasma, and monitor the blood concentration of critically ill patients. METHODS The determination was performed on Diamonsil C₁₈ column (4.6 mm×250 mm, 5 μm) with chloramphenicol as internal standard, acidified acetonitrile as protein precipitator, and bisolvent mobile phase consisting of A acetonitrile, B citric acid (0.1 mol·L^-1)-sodium hydrogen phosphate (0.2 mol·L^-1) buffer solution (pH 3.0). The flow rate was 0.5 mL·min^-1, the detection wavelength was set at 254 nm, the column temperature was maintained at 30 ℃, and the sample size was 20 μL. RESULTS Linezolid had a good linear relationship in the concentration range of 0.5-40 μg·mL^-1 (r=0.999 9). The lower limit of quantitation was 0.5 μg·mL^-1, and the detection limit was 0.1 μg·mL^-1. PNU-142300 and PNU-142586 had good linear relationship in the concentration range of 0.5-20 μg·mL^-1 (r=0.999 8 and r=0.999 6), the lower limit of quantitation was 0.5 μg·mL^-1, and the detection limit was 0.2 μg·mL^-1. In 10 critically ill patients, plasma linezolid and PNU-142300 and PNU-142586 were 3.62, 1.88 and 2.30 times higher in patients with renal insufficiency than in patients with normal renal function, respectively. There was a strong correlation between exposure of linezolid and its metabolites and thrombocytopenia and anemia. CONCLUSION The method is simple, sensitive, and accurate, and can be used to monitor the blood concentration of linezolid and its metabolites PNU-142300 and PNU-142586 in critically ill patients and study the correlation of adverse reactions.