Natural drugs have been used in clinic for thousands of years in China, with complete and systematic theories, and are the treasure of the Chinese nation. Among them, flavonoids, polysaccharides, iridoid glycosides, saponins and other natural drugs have been proved to have curative effects on many diseases. Generally, the bioavailability of natural drugs absorbed in the intestine after oral administration is relatively low. As one of the ecosystems regulating the host's adaptation to the environment, the gut microbiota has a two-way effect on the host's internal and external environment, which is usually closely related to the efficacy of natural drugs. In order to understand the mechanism of the effect of drugs that are difficult to absorb, the exploration of targeting the gut microbiota has become a new strategy in recent years. The metabolism of natural drugs is often achieved through the enzymes produced by the gut microbiota, which is closely related to its efficacy and safety. The research of metabolic transformation by the gut microbiota is still limited. So in this review, we mainly discussed the characteristics of major metabolic enzymes derived from gut microbiota and the research progress in the metabolism and transformation of natural drugs, aiming to enrich the information on the metabolism mechanism of natural drugs under the action of gut microbiota and provide evidence for proving that gut microbiota is part of the regulation axis of other organs. It also provides some scientific reference for deepening the understanding and cognition of the metabolism and transformation of natural drugs under the action of intestinal bacteria.

Matricaria chamomilla L. is a herbaceous plant belonging to Asteraceae family. It is frequently used as medicinal herb in the Uygur medicines and is mainly produced in Xinjiang Uygur autonomous region, Europe and other places. Its chemical constituents mainly include flavonoids, phenolic acids, essential oil, coumarins and other compounds. This herb has anti-inflammatory, analgesic, antibacterial, antioxidant, anti-tumor, hypoglycemic, hypolipidemic, hypotensive and pharmacological effects on gastrointestinal and nervous system. Based on the overview of the chemical constituents and pharmacological effects of chamomile, this paper conducts a predictive analysis of its quality markers. It is speculated that flavonoids, phenolic acids and sesquiterpenoids can be used as quality markers of chamomile, providing scientific reference for establishing its quality standards.

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 an effective identification method for Jiawa and easily mixed species by using DNA barcoding ITS2 sequence. METHODS The total DNA of 72 Jiawa and its similar species samples were extracted by using the plant genomic DNA Extraction kit,PCR amplification and bidirectional sequencing were carried out for the ITS2 fragments, another 30 items of the genus were downloaded from GenBank. MEGA6.0 software was applied to analyze 102 ITS2 sequences, calculate the intraspecific and interspecific K2-P distance, construct neighber-jioning (N-J) phylogenetic tree, and predict the secondary structure of ITS2. RESULTS K2-P genetic distance analysis showed that, in addition to Pleurospermum franchetianum Hemsl. and Pleurospermum wrightianum de Boiss., Pleurospermum hookeri var. thomsonii C.B. Clarke., Sphallerocarpus gracilis (Bess.) K.-Pol., Vicatia thibetica de Boiss., Carum carvi L., Heracleum millefolium Diels., Ligusticum daucoides (Franch.) Franch. and Anthriscus sylvestris subsp. Nemorosa have obvious barcode spacing. N-J tree cluster analysis showed that each species could be clustered into one branch with a bootstrap support rate of >50%, and the monophyleticity was good. The secondary structure analysis of ITS2 showed that the number, size and position of stem-loops in the four helical regions of Jiawa and its miscible were different. CONCLUSION The ITS2 sequence can be used to identify Jiawa and its easily mixed species, which is beneficial to the rational development and utilization of Jiawa resources.

OBJECTIVE To research and evaluate risk of 18 polycyclic aromatic hydrocarbons(PAHs) in seed-fruit herbs based on a SPE-isotope dilution GC-MS/MS method for the determination of the residues. METHODS Using isotope as internal standard, the sample was extracted by n-hexane and purified by molecular imprinting solid phase extraction. GC-MS /MS method was used for the assay. The chromatographic column was DB-17ms(0.25 mm×30 m, 0.25 μm) with temperature programming and MRM detection. The preliminary risk assessment of polycyclic aromatic hydrocarbons (PAHs) in seeds and fruits of traditional Chinese medicine (TCM) was carried out by using toxicity equivalent factor method. RESULTS The calibration curves for the 18 kinds of typical PAHs were linear in the range of 2-50 ng·mL^-1. The average recovery rate was in the range of 77.25%-112.32%, with RSD 2.74%-15.89% (n=3). The LOQs were 0.2-1 μg·kg^-1 According to the risk assessment results, some varieties may have potential cancer risk. CONCLUSION This method is specific, sensitive, and can be used for residue detection of 18 typical kinds of PAHs in seed-fruit herbs. The residual amount of PAHs in some seeds and fruits of traditional Chinese medicine may have a potential cancer risk, which need attention.

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.