OBJECTIVE To preparet aurocholic acid modified PLGA nanospheres for oral delivery of semaglutide. METHODS The nanospheres were prepared by dobule emulsion solvent evaporation technique, and the preparation process was optimized by single-factor experiments; the nanospheres were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and laser particle size measurement; pharmacokinetic experiments were performed using SD rats; pharmacodynamic experiments were performed using db/db mice. RESULTS The FT-IR showed that taurocholic acid was successfully modified to the surface of the nanospheres. The particle size of the nanospheres was (185.9±3.31) nm, the ζ-potential was (-32.53±0.95) mV, and the drug loading and encapsulation rates were (11.15±0.07)% and (85. 51±0.01)%. The nanospheres showed good sustained release in vitro, with a cumulative release rate of 84. 96% within 192 h. Pharmacokinetic experiments were performed in SD rats, and the results showed that the bioavailability of nanospheres was 2.5%, and the slow release of semaglutide could be achieved within 192 h. The efficacy of nanospheres was verified in db/db mice, and the results showed that after gavage administration of nanospheres, the blood glucose of diabetic mice decreased rapidly and remained stable for about 3 d. CONCLUSION The oral delivery of semaglutide nanospheres prepared in this study has high drug loading and encapsulation efficiency, which can effectively control the blood glucose of diabetic mice within 3 d and improve the bioavailability.

OBJECTIVE To synthesize scutellarein derivatives and study their solubility and bioactivity. METHODS Based on the lead compound scutellarein, 12 compounds were designed and synthesized using functional group substitution, parent nucleus ring opening, and electron rearrangement principles. The structures of the compounds were characterized through ¹H-NMR, ¹³C-NMR and MS, their solubility was tested, and their transient receptor potential vanilloid 3 (TRPV3) inhibitory activities were evaluated through calcium flow detection. An animal model of psoriasis induced by imiquimod (IMQ) was established and the three most active compounds were selected for anti-psoriasis evaluation. RESULTS All compounds had higher solubility than scutellarein. In the calcium flow experiment, compounds I-01, I-04, and I-06 showed stronger TRPV3 inhibition than scutellarein, with I-06 having the strongest activity. Animal experiments on psoriasis showed that I-01, I-04, and I-06 could significantly improve the pathological appearance and inflammatory response of psoriasis mice, and I-06 performed the best. CONCLUSION This type of modified scutellarein has potential therapeutic value for psoriasis and provides a new approach for targeting TRPV3 to treat various skin diseases.

OBJECTIVE To analyze the dynamic changes of cell types and protein expression during the occurrence and development of ulcerative colitis. METHODS Single-cell sequencing technology was used to analyze the intestinal tissues of ulcerative colitis model mice and normal mice. After quality control, a total of 58 714 cells were used to construct a colitis dataset for downstream analysis. Then, the changes of epithelial compartments in the process of colitis occurrence and development were analyzed by means of dimensionality reduction clustering, GO analysis and Monocle quasi-time analysis. RESULTS Compared with the normal samples, the interaction between cell subsets in colitis samples changed significantly. The analysis revealed that most epithelial cell subtypes were destroyed and reduced in number during the colitis phase. The Wnt pathway was inhibited, which is important for maintaining stem cell function. The Ppia-Bsg ligand receptor pair was highly expressed in normal samples. CONCLUSION The depletion of Ppia secreted by stromal cells in the inflammatory state leads to stem cell senescence, which ultimately disrupts the homeostasis of epithelial cells and aggravates colitis.

It is important to select a suitable in vitro experimental model to study the pathology and pharmacological mechanisms of inflammatory bowel disease (IBD), thereby developing advanced therapeutic drugs. Immortalized cell lines, as a classic in vitro model, offer several advantages in the research and drug evaluation of IBD, such as high efficiency, low cost, simple operation and intuitive experimental results. However, these models cannot recapitulate the multicellular composition and intercellular interaction of intestinal tissue in vivo and may lose genetic characteristics after multiple passages in vitro. The development of organoids and organs-on-a-chip has promoted the technological innovation of in vitro models by significantly improving our capability to simulate the architecture and function of IBD. The enhanced reproducibility of the organoid model to the microenvironment of the source tissue also confers improved predictive capability for patient treatment response. This paper reviews the current research status of these models by discussing their characteristics, advantages, disadvantages, and their applications in evaluation of IBD therapeutic drugs and development of advanced therapeutic drugs, as well as in the exploration of pharmacological mechanisms.

OBJECTIVE To prepare risedronate sodium (RIS)-loaded dissolving microneedle (DMN) and evaluate its efficacy in preventing postmenopausal osteoporosis. METHODS The preparation process of RIS-DMN was optimized by Box-Behnken design of response surface methodology. The appearance, solubility, mechanical property, safety and transdermal effect of RIS-DMN were characterized by scanning electron microscope, intradermal dissolution test, puncture test, skin barrier recovery test and in vitro permeation test. The pharmacodynamic evaluation of RIS-DMN was performed in ovariectomized osteoporosis model rats. RESULTS The optimal formulation were determined to be 45% for solute (mixed with 1:0.86 PVP K30 and CS) and 55% for solvent. It was found that the RIS-DMN have good physical characteristics and properties, and showed great effects in regulating the level of Ca²⁺, P³⁺and alkaline phosphatase (ALP). Meanwhile, the RIS-DMN showed great effects in repairing bone microstructure and improving bone density in ovariectomized osteoporosis model rats, as similar as oral administration. CONCLUSION The RIS-DMN has stable quality, convenient use and precise efficacy, shows great potential in the treatment of postmenopausal osteoporosis.

OBJECTIVE To develop the limited standard and supporting detection methods of commonly used pesticides in Lycium barbarum L. (L. barbarum). METHODS In previous research, the “the Principle of Conversion of Limit Standards of Traditional Chinese Medicine in <GB 2763 national food safety standard-maximum residue limits for pesticides in food>” have been drafted. In this research, the L. barbarum was used as the research object to study the conversion scope, risk assessment, detection methods and conversion judgment principle. RESULTS A method for the determination of 13 pesticide residues in the L. barbarum was established by using acetonitrile extraction and HLB column purification. And the GC-MS/MS and LC-MS/MS technology were used to detect the pesticide residues. In addition, the maximum residue limit of 13 pesticides in L. barbarum was finally formulated by sample determination. CONCLUSION The specific implementation steps are provided for the conversion of GB 2763 standard to traditional Chinese medicine standard in this study, and it is of great significance for promoting the connection between planting and circulation supervision of L. barbarum.

OBJECTIVE To investigate the ameliorative effect of Guifu Dihuang Wan (GFDHW) on intestinal lipid absorption dysfunction in aging mice and its potential mechanisms. METHODS Twenty-six SPF male C57BL/6J mice aged 11 months were raised to 21 months and divided into an aging model group (Model group) and a GFDHW group (1.17 g·kg^-1). The GFDHW group received medication via feed administration for 3 months, with free access to water during the administration period, while the Model group was fed normal feed and had free access to water. Weekly changes in body weight were recorded for each group of mice, and specimens were collected at 24 months of age. Nine SPF male C57BL/6J mice aged 8 months were used as the youth control group (Con group) and were acclimatized for 1 week before specimen collection. Prior to specimen collection, all groups of mice were fasted for 24 hours, and 0.2 mL of olive oil was administered by gavage to each group of mice at the end of the fasting period, followed by gavage of 0.2 mL of semi-solid nutrient solution 30 minutes before specimen collection. Small intestine propulsion tests were conducted to assess the small intestine motility of each group of mice. Biochemical reagent kits were used to measure the serum levels of triglycerides (TG), total cholesterol (TC), and low-density lipoprotein (LDL) in each group of mice. Transmission electron microscopy was used to observe changes in the number of lipid droplets in intestinal epithelial cells of each group. The calcium cobalt method was used for alkaline phosphatase staining to detect intestinal alkaline phosphatase (IAP) activity in each group of intestines. Hematoxylin-eosin (HE) staining was used to observe the morphology of intestinal epithelia in each group. Immunohistochemical staining was performed to detect the expression of Olfm4, Lgr5, PCNA, Notch1, Hes1 and Villin in the small intestines of each group of mice. Western blot analysis was conducted to determine the expression levels of lipid absorption-related molecular proteins in the small intestines of each group of mice. RESULTS Compared with the Con group mice, the Model group mice exhibited faster weight loss, decreased small intestine motility, increased serum TG, TC, and LDL levels, and increased number of lipid droplets in intestinal epithelial cells. The GFDHW group mice showed improved small intestine motility and decreased serum TG, TC, and LDL levels. HE staining revealed disordered arrangement and fracture of small intestinal villi, shortened villus length, decreased crypt depth, and reduced crypt numbers in Model group mice, whereas the GFDHW group mice exhibited orderly and continuous arrangement of small intestinal villi, increased villus length, higher crypt depth, and increased crypt numbers. The immunohistochemical results showed that compared to the Con group mice, the expression of Olfm4, PCNA, Notch1, Hes1, and Villin proteins in the small intestine of the Model group mice decreased, while the levels of these proteins increased in the GFDHW treated group mice. Western blot analysis revealed that compared to the Con group mice, the expression of CD36, FATP4, SR-BI, ACAT2, FABP1 and MTTP proteins in the small intestine of the Model group mice decreased, whereas the levels of these proteins increased in the GFDHW treated group mice. CONCLUSION GFDHW improves lipid absorption function in aging mice by upregulating Notch1 expression, promoting differentiation of intestinal stem cells into absorptive cells, and increasing the expression levels of related molecular proteins in the lipid absorption pathway.

OBJECTIVE To carry out screening of pesticide residues in Panax notoginseng for the pesticide residues commonly used in Panax notoginseng,understand the pesticide residues situation and carry out the related risk assenssment study. METHODS GC-MS/MS and LC-MS/MS methods were used to establish detection methods for commonly used pesticides in Panax notoginseng, as well as those regulated by laws and regulations; the methods were used to conduct a comprehensive screening and risk assessment of the collected Panax notoginseng samples; and the indexes of the transformed pesticides were confirmed in accordance with the pesticide residue limits of Panax notoginseng in the GB 2763 standard. RESULTS In this study, the residue determination methods for about 40 commonly used pesticides and pesticides regulated by regulations in Panax notoginseng were successfully established, and the corresponding limits were set according to the GB 2763 standard. Eight pesticides including tebuconazole, phenyl ether metronidazole and carbendazim were finally recognized as transformed pesticide indicators. CONCLUSION The pesticide residue detection method for Panax notoginseng established in this study is characterized by simple and rapid operation, high specificity and high sensitivity, and the related risk assessment and limit setting provide technical support for the improvement of the quality standard of Panax notoginseng.

OBJECTIVE To establish a quantitative nuclear magnetic resonance coupled with high performance liquid chromatography (qNMR-HPLC) technique for the rapid determination of cilostazol impurity Ⅰ correction factor. METHODS The mixture of cilostazol and cilostazol impurity Ⅰ was dissolved in deuterated dimethyl sulfoxide. A portion of the solution was determined by qNMR, while the other portion of the solution was diluted with water-acetonitrile (60:40) and analyzed by HPLC. The correction factor of cilostazol impurity Ⅰ was calculated with the response signals from qNMR and the peak areas from HPLC. The correction factor of cilostazol impurity Ⅰ was also determined by HPLC standard curve method. A mixed solution containing residual solvent was prepared to simulate the effect of solvents in determining correction factors. When the content of cilostazol impurity Ⅰ was inaccurately assigned due to residual solvent, difference between qNMR-HPLC method and standard curve method was compared. RESULTS When the contents of cilostazol and cilostazol impurity Ⅰ were assigned accurately, the correction factors for cilostazol impurity Ⅰ by qNMR-HPLC method and HPLC standard curve method were 1.74 and 1.76, respectively, which were basically consistent with the pharmacopoeial results. When cilostazol impurity Ⅰ contained residual solvents, and there was an error in the content assignment, the correction factor of the determination by the qNMR-HPLC technique was still 1.72, and the result was not affected by the accuracy of the content. While the correction factor of HPLC standard curve was 2.01, which was deviated from the actual results. CONCLUSION Compared with the HPLC standard curve method, the qNMR-HPLC coupling technique is independent of the accuracy of the content of the substance to be measured and the weighing volume, and does not require purification to prepare a high purity compound. qNMR-HPLC is a powerful tool in the determination of impurity correction factors.

OBJECTIVE To establish an analysis method of the charge heterogeneity of human urinary kininogenase (HUK) by using image capillary isoelectric focusing (iCIEF) and complete the methodological verification. METHODS The 35 μL 1% methyl cellulose, 10 μL 200 mmol·L^-1 IDA, 4 μL pharmalyte electrolyte, 48 mg urea, 0.5 μL pI marker with pI 6.15 and 7.05 were added in the sample solution. The focusing condition was pre-focusing voltage 1 500V, duration 1 min, focusing voltage 3 000 V, duration 6 min. RESULTS The optimized method has stable baseline, and the target protein was significantly different from the unrelated protein, The recovery rate of accuracy verification was within 90%-110%, and the linearity verification result had r² of 0.995 7, The RSD of each isomer pI in the repeatability verification was less than 0.2%. The limit of quantification was 0.013 mg·mL^-1. The concentration of urea durability, IDA durability and the electrolyte pharmalyte durability are good. Using this method, the charge isomers of HUK from different manufactures were analyzed effectively. CONCLUSION The developed iCIEF method has good specificity, precision, linearity and durability, and can solve the problem of charge heterogeneity evaluation of protein products, which is of great significance to the quality control of such products from the perspective of charge heterogeneity.