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 screen and study the effects of small activating RNA (saRNA) enhancing the expression of ABCG2 gene on renal and intestinal cell excretion of uric acid (UA) to explore a new type of therapeutic mechanism drugs to promote UA excretion. METHODS saRNAs targeting mice and human ABCG2 genes were designed and screened by real-time quantitative reverse transcription PCR(RT-qPCR) and Western blot in mice TCMK-1 and human HK-2 cells. The effect of the selected saRNA on the excretion of UA was examined in TCMK-1 and HK-2 cells. The mice saRNA was injected into the tail vein of hyperuricemia (HUA) model mice. The levels of serum uric acid (SUA), urinary uric acid (UUA), intestinal uric acid, blood urea nitrogen (BUN) and serum creatinine (SCr), and the activity of liver xanthine oxidase (XOD) in the mice were detected to verify the ability of selected saRNA to promote UA excretion. The pathological changes of kidney and small intestine were analyzed through H&E. The expression of ABCG2 in kidney and small intestine was analyzed by RT-qPCR, Western blot and immunohistochemistry. RESULTS Four saRNAs targeting ABCG2 were selected, 2 for mice and 2 for human, named saRNA-1/2 and hsaRNA-1/2 respectively, which increased the expression of ABCG2 in TCMK-1 and HK-2 cells (P<0.05), and promoted the extracellular excretion of UA (P<0.05). In HUA model mice, saRNA-1/2 increased the levels of UUA and intestinal UA (P<0.05), reduced the levels of SUA, BUN, and SCr (P<0.05), decreased the damage of HUA to kidney and small intestine, and enhanced the expression of ABCG2 in kidney and small intestine, but did not show significant effect on the activity of liver XOD (P>0.05). CONCLUSION The selected saRNAs could enhance the expression of ABCG2 and promote the excretion of UA in cells and mice, reducing SUA level.

OBJECTIVE To prepare berberinehydrochloride resincomplex for good taste-masking effectof lyophilized oraldisintegration tablets using berberine hydrochloride as a model drug and using Kyron-T114 as the drug-loaded resin. METHODS The drug-resin composites were prepared by using the static method. Then drawn the adsorption isotherm and adsorption kinetic curves to study the adsorption mechanism of the resin. Afterwards, the characteristic of the berberine hydrochloride resin complex was studied by using the SEM, DSC, XRD and FTIR. Finally, prepared the oral disintegration tablets of berberine hydrochloride and its resin complex by using the lyophilized extrusion technology. Next, studied the in vitro release experiments and evaluated the effect of masking taste. RESULTS According to the research, a berberine hydrochloride-resin complex with the drug loaded of 43.04% was prepared. And the analysis showed that the adsorption of the drug on the resin was the multilayered chemical adsorption. The dissolution of berberine hydrochloride's powder is mainly affected by the solubility of berberine hydrochloride. Moreover, the dissolution mechanism is mainly based on Fick diffusion. The dissolution of berberine hydrochloride's powder is mainly affected by the pH of the solution, along with the concentration and the type of the ion in the solution. In artificial gastric juice, the drug dissociates rapidly from the resin. So the film diffusion was the rate-limiting step for dissolution. On the contrary, the drug dissociated slowly in the water, artificial saliva, artificial intestinal fluids including pH 4.5 and pH 6.8. So the particle diffusion was the rate-limiting step for dissolution. Besides, the dissolution of lyophilized oral disintegration tablets of drug-resin complexes in artificial saliva was significantly lower than that of berberine hydrochloride oral disintegration tablets. Lastly, the results from sensory evaluation and electronic tongue detection proved that the lyophilized oral disintegration tablets of berberine hydrochloride-resin compound has good taste masking effect. CONCLUSION Berberine hydrochloride resin complex lyophilized mouth collapse has good taste masking effect. Moreover, the drug-resin complex prepared by Kyron-T114, a weak cation exchange resin, is beneficial to improve the dissolution of poorly soluble drugs in artificial gastric juice, thereby improving their oral bioavailability. Thus, it can provide a new idea for the taste masking of traditional Chinese medicine.

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.

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 and analyze the medical information of anti-hypertensive drugs instructions in children and provide reference for clinical rational drug. METHODS A total of 473 instructions of anti-hypertensive drugs were collected. The determine standards for the dosage and medical information guidance of anti-hypertensive drugs in children were established according to Provisions on the Administration of Drug Instructions and Labels, and then the labeling of medical information in children was analyzed. RESULTS The dosage of anti-hypertensive drugs in children was labeled 55.18%, vaguely labeled 5.50% and unlabeled 39.32%, and then the medical guidance information of anti-hypertensive drugs in children was labeled 31.50%, vaguely labeled 28.54% and unlabeled 39.96%. CONCLUSION The information labeling rate of anti-hypertensive drugs in children is low, the medical information is insufficient, and there is still a certain risk of medication in children.

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.

Chemotherapy is one of the commonly used methods for tumor treatment, but the occurrence of multidrug resistance (MDR) in tumors has added great difficulties for tumor treatment, causing serious harm to patients' physiology and psychology. At present, there is a relative lack of research and treatment on tumor resistance in clinical practice, and finding reversal agents and strategies for tumor resistance is an urgent problem to be solved. This article reviews the mechanism of tumor multidrug resistance and its reversal methods and research status of Chinese medicine as reverse agent, aiming to provide reference for future research.

OBJECTIVE To evaluate the microbial contamination levels of 14 kinds of mold-prone TCM decoction pieces and the diversity of fungi on the surface. METHODS The total aerobic microbial count (TAMC), total yeasts and molds count (TYMC), heat-resistant microbial count (HRMC), and three types of control bacteria of mold-prone TCM decoction pieces were analyzed by referring the Chinese Pharmacopoeia 2020 Edition Volume Ⅳ 1108 microbiological limit test method. Furthermore, the fungi on the surface were cultivated using the plate method, and the single strain was isolated by the top purification method, which was further identified by colony morphology and molecular identification. RESULTS The results showed that TAMC of sample ranged 250-2.45×10⁶ cfu·g^-1, TYMC ranged 5-6.6×10⁵ cfu·g^-1, and HRMC ranged 0-30 cfu·g^-1. The remaining samples of cholesterol-resistant progestin-negative bacteria did not reach 1×10⁴ cfu·g^-1, with the exception of one batch of Polygalae Radix and one batch of Glycyrrhizae Radix et Rhizoma. Although only one batch of Citri Reticulatae Pericarpium tested Escherichia coli and no Salmonella was detected in all samples, significant conditional pathogenic bacteria such as Enterobacter cloacae, Klebsiella pneumoniae, and Cronobacter sakazakii were found in the control bacterial examination items. ITS sequence sequencing comparison identified 106 strains in total, grouped into 16 genera, with Aspergillus, Penicillium, and Cladosporium accounting for 31%, 18%, and 17%, respectively, of the major bacteria. Among the genus Aspergillus, 11 strains of A. niger, 7 strains of A. flavus, 2 strains of A. fumigatus, and 1 strain of A. miscellaneous were isolated as pathogenic fungi. CONCLUSION It can provide early risk warning for its potential fungi toxins contamination through research on microbial pollution and surface fungi diversity.

OBJECTIVE To investigate the effects of carbohydrate components of fungus S7 (Tulasnella sp.) on seed germination of Dendrobium officinale. METHODS The polysaccharides of S7 mycelia (MP) and of S7 fermentation broth (FP) were prepared by water extraction and alcohol precipitation. MP and FP were hydrolyzed by trifluoroacetic acid to obtain hydrolysates, MPH and FPH. The monosaccharide composition of MP and FP were determined by PMP-HPLC. MP, FP, MPH, FPH, and the unique and mixed monosaccharide of MP were added to water agar (WA) and 1/2 MS media respectively to study the effects of carbohydrate on the seed germination of Dendrobium officinale. RESULTS The germination rate of WA medium group (WACK1) was (88.65±4.71)%, and seeds germinated to stage 3 (protocorm). Compared with WACK1, adding MP and MPH had no significant effect on germination rate (P>0.05), and seeds could germinate to stage 4 (having one leaf), while adding FPH could significantly reduce germination rate (P<0.05). MP was composed of glucose, galactose, glucuronic acid, xylose, mannose, fucose, ribose and arabinose in a molar ratio of 15.05∶1.00∶0.70∶0.35∶0.30∶0.29∶0.13∶0.13. FP was composed of arabinose, galactose, xylose and glucuronic acid in a molar ration of 1.22∶1.00∶0.71∶0.47. Fucose was the unique monosaccharide component of MP. The germination rate and stage 5 (having two leaves) germination rate of 1/2 MS medium group(MSCK) was (98.10±0.46)% and (50.97±4.33)%, respectively. Compared with MSCK, adding 0.20-20 mg·L^-1 fucose had no significant effect on germination rate (P>0.05), while significantly increased stage 5 germination rate (P<0.05), which was 1.36-1.47 times that of MSCK. CONCLUSION MP and MPH promote the development of protocorm of D.officinale. Fucose is an active component in MPH that could promote the further differentiation and development of seeds after germination.