OBJECTIVE To investigate the feasibility of performing identification of Citri Sarcodactylis Fructus (CSF) and Citri Fructus (CF) based on high performance liquid chromatography (HPLC) fingerprint coupled with multivariate data analysis. METHODS HPLC method was applied to establish the fingerprints of CSF and CF. Similarity evaluation of 12 batches of CSF samples and 10 batches of CF samples was performed by Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2004A edition). Hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used for data analysis. RESULTS HPLC fingerprints of CSF and CF were established, respectively. The fingerprint similarities of CSF samples, as well as the fingerprint similarities of CF samples, were all greater than 0.9. The similarities between the fingerprints of CSF samples and the control fingerprint of CF, as well as the similarities between the fingerprints of CF samples and the control fingerprint of CSF, were all less than 0.9, which indicated significant differences in chemical composition between CSF and CF. The total 22 samples were divided into two groups by HCA, PCA and OPLS-DA, which was consistent with the two varieties. Three differential markers were screened by OPLS-DA and two of them were identified as 5, 7-dimethoxycoumarin and hesperidin by reference substances. The statistical analysis indicated that the ratio of absolute peak area (APA) of hesperidin and 5, 7-dimethoxycoumarin can distinguish CSF and CF from each other simply and accurately. CONCLUSION HPLC fingerprint coupled with multivariate data analysis can be used to identify CSF and CF.

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 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 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.

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 investigate the efficacy and safety of different preparations of intravenous immunoglobulin (IVIG) for Kawasaki disease treatment in children. METHODS A retrospective analysis of medical records of pediatric Kawasaki disease patients at our institution in recent three years was conducted. Patients were divided into preparation A group and preparation B group based on the type of IVIG preparations used. Both groups received IVIG (2 g·kg^-1) in conjunction with aspirin therapy following Kawasaki disease diagnosis. Changes in body temperature, white blood cell count, platelet count, C-reactive protein levels, IVIG cost, length of hospital stay, and adverse events were compared. RESULTS According to the drug labeling, there are certain differences between preparation A and preparation B. Preparation B exhibited a significantly shorter duration of fever resolution compared with preparation A (P<0.005), and at the first temperature test after infusion, the body temperature of preparation B group was significantly lower than that of preparation A group (P<0.05). Furthermore, preparation B demonstrated a more significant reduction in white blood cell count following treatment (P=0.010). In terms of hospital stay duration and IVIG monotherapy costs, preparation B had significantly shorter hospital stay (P<0.05) and lower per-unit IVIG expense (P<0.001). No significant differences were observed between the two groups in terms of adverse events, including rashes, infusion interruptions due to high fever, liver injury, and coronary dilation. CONCLUSION There is significant difference in efficacy between different preparations of intravenous immunoglobulin in the treatment of Kawasaki disease in children, but the safety is similar.

Thrombocytopenia is a disorder characterized by a decreased platelet count in the peripheral blood, which can be caused by various congenital and acquired diseases. Currently, thrombopoiesis-promoting drugs used for thrombocytopenia primarily include recombinant human interleukin-11 (rhIL-11), recombinant human thrombopoietin (rhTPO), and thrombopoietin receptor agonist (TPO-RA). Hetrombopag is an innovative oral non-peptide TPO-RA that has been approved for treating chronic adult primary immune thrombocytopenia (ITP) with inadequate response to glucocorticoids and immunoglobulin, as well as severe aplastic anemia (SAA) with poor response to immunosuppressive therapy in China. Clinical studies have demonstrated that hetrombopag is safe and well-tolerated in ITP and SAA patients. Currently, there is an ongoing phase Ⅲ clinical trial investigating the efficacy of hetrombopag in managing chemotherapy-induced thrombocytopenia (CIT). This article provides a review on the research progress of hetrombopag in treating thrombocytopenia, aiming to offer valuable insights for clinical practice.

Quercetin, a natural polyphenol, is one of the most abundant flavonoids found in many plant foods. Quercetin is a natural antioxidant, which can protect cells from damage caused by free radicals. Quercetin is a lipophilic compound that can pass through the cell membrane and regulate many intracellular and extracellular signal pathways related to disease progression and chemoprevention. Quercetin has a wide range of pharmacological activities, anti-inflammatory, antibacterial, anti-cancer and prevention of cardiovascular and cerebrovascular diseases, but its low solubility and bioavailability limit its clinical application. In order to improve these drawbacks, people have carried out structural modifications on quercetin, such as modifying hydroxyl groups to generate esters or ethers, modifying carbonyl groups to generate hydrazone or carbonyl oxygen substituted products, 8-substituted products, and forming complexes with metal ions. Quercetin derivatives with good solubility, high bioavailability, improved activity and enhanced anti-cancer activity were obtained by optimizing the structure of quercetin. The research progress of quercetin derivatives in recent years is reviewed to provide reference for the further development of quercetin derivatives.

OBJECTIVE To find new arylaminoquinazoline-based antitumor target compounds with high efficacy and low toxicity. METHODS A series of N-phenyl-4-(trifluoromethyl)quinazoline-2-amine derivatives were synthesized starting from 2-nitrobenzaldehyde or 6-nitroveratraldehyde by a combination of nucleophilic addition, oxidation, reduction, cyclization, chlorination, and coupling reactions. The structures of the target compounds were characterized using NMR and MS. Their antitumor activities in vitro were then evaluated against four human cancer cell lines (PC3, LNCaP, K562, and HeLa) using MTT assay. The target prediction of active compound 8b was performed. RESULTS Compounds 8b and 8c exhibited promising anti-proliferative properties, particularly compound 8b, which exhibited excellent antitumor activity against PC3, LNCaP, and K562 with IC₅₀ values of 5.51, 4.51 and 8.49 μmol·L^-1, respectively. Molecular docking experiment demonstrated that PIM-1 is the potential target of 8b. CONCLUSION The piperazine ring containing compound 8b exhibits the strongest antitumor activity, which is worthy of further study.

OBJECTIVE To design and synthesize a series of bifunctional ruthenium complexes containing hydroxamic acid as HDAC6 selective inhibitors by conjugating aromatic hydroxamic acid with bipyridine ruthenium (Ⅱ) complexes, and investigate the in vitro antitumor activity. METHODS Three ruthenium complexes were synthesized with aromatic ring as ‘Linker’ and hydroxamic acid as zinc binding group(ZBG), and their structures were characterized by ¹H-NMR, ¹³C-NMR and HRMS spectrometry. The HDAC inhibitory activity was evaluated by fluorescence analysis. The in vitro antitumor activities against A549, MDA-MB-231, MCF-7, HepG-2 and LO2 cell lines were evaluated by MTT assay. The binding of compounds to the active site of HDAC6 protein was studied by molecular docking. RESULTS All compounds showed selective HDAC6 inhibitory effect, in vitro antitumor activity and tumor-targeting activity, among which representative compound 3 exhibited comparable cytotoxic activity to cisplatin and much higher tumor-targeting activity than cisplatin. CONCLUSION The introduction of a wider “Cap” (surface recognition unit) in the pharmacophore model can improve the specific recognition of the compound against HDAC6, which proved that the design of bifunctional aromatic hydroxamic acid and bipyridine ruthenium complexes is rational.