Bioaffinity monolithic chromatography is a chromatographic technology developed by combining the latest advances in biology, hylology and chemical analysis. Based on the basic principle of biological affinity, monolithic materials were used to prepare stationary phase, which can simulate intermolecular interactions in vivo to screen and purify active substances. This technology can specifically customize analysis methods for different target active substances and has been widely used in multiple research fields in pharmacy, chemistry and biology. This article focuses on the preparation methods of monolithic columns, systematically reviews the selection for chromatographic columns, as well as the strategies for affinity ligand immobilization such as covalent binding of biomolecules and biological labels. It also summarizes the relevant applications and development directions of bioaffinity monolithic columns in the preparation of active proteins and drug/biological analysis.

Platycodonis Radix is a traditional Chinese medicine as homology of medicine and food, and it has the functions of clearing pharynx, moistening throat, regulating immune system, assisting in lowering blood sugar and blood lipid. Platycodonis Radix contains saponins, flavonoids, phenols, polyacetylenes, polysaccharides and other chemical components. Modern pharmacological studies have demonstrated that Platycodonis Radix has anti-tumor, anti-oxidation, anti-obesity, anti-diabetes and other biological activities. In addition, Platycodonis Radix has high value in medicinal and healthcare, so it is of great significance to strengthen the development of Platycodonis Radix healthy products. This paper summarizes the research progress of Platycodonis Radix from three aspects as chemical composition, pharmacological action and modern application, which can provide some valuable references for the comprehensive utilization and development of Platycodonis Radix.

OBJECTIVE To optimize the processing technology of salt Anemarrhenae Rhizoma, and to clarify the correlation between the chemical composition content and color value of salt Anemarrhenae Rhizoma. METHODS Mangiferin, neomangiferin, isomangiferin, timosaponin AⅡ, timosaponin AⅢ, timosaponin BⅡ and timosaponin BⅢ were used as evaluation indexes, and the amount of salt water, stir-frying time and stir-frying temperature were used as influencing factors. Design-Expert 12.0 software, Box-Behnken response surface method combined with weighting method were used to optimize the processing technology of salt Anemarrhena Rhizoma. The chromaticity values of 17 samples with different process parameters under response surface method were determined, and Pearson correlation analysis was performed on their components and chromaticity values. RESULTS The optimum processing technology was as follows:50 mL of 80 mg·mL^-1 salt water was added to each 200 g of Anemarrhenae Rhizoma decoction pieces, the stir-frying time was 15 min, and the stir-frying temperature was 140 ℃. The results of chromaticity study showed that the content of mangiferin, neomangiferin, isomangiferin and timosaponin BⅡ was negatively correlated with the chromaticity value L*, and positively correlated with the chromaticity values a* and b*, indicating that the smaller the L* value, the larger the a* value, the larger the b* value, the closer the salt to the dark yellow, the higher the content of mangiferin, neomangiferin, isomangiferin and timosaponin BⅡ. CONCLUSION The optimized processing technology is stable and feasible. The analysis method for the determination of 7 components in salt Anemarrhenae Rhizoma is accurate and reliable. The chemical composition of salt Anemarrhenae Rhizoma has a significant correlation with the color value. The quality of salt Anemarrhenae Rhizoma can be preliminarily evaluated by the appearance color, which provides a theoretical basis for its multi-dimensional quality evaluation.

OBJECTIVE To identify the chemical constituents in flower of Fritillaria unibracteata P. K. Hsiao et K. C. Hsia (F-F, Chinese: Anzibeimu Hua) by employing ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS) for the optimal utilization of Fritillaria cirrhosa D. Don (Chinese: Chuanbeimu) resources. METHODS The separation was conducted on Thermo Hypersil GOLD aQ column (2.1 mm×150 mm, 1.9 μm) eluted with methanol-0.1% formic acid in a gradient mode at a flow rate of 0.3 mL·min^-1, the column temperature was maintained at 35 ℃, and positive and negative electrospray ionization(ESI) was adopted for determine the chromatographic effluents. RESULTS Through comparison with reference standards, precise molecular weight determination from primary mass spectrometry, and analysis of fragmentation patterns from secondary mass spectrometry, combined with existing literature, 71 chemical constituents were identified in F-F. These identified compounds encompass 24 alkaloids, 12 flavonoids, 9 amino acids, and additional substances, including the notable detection of peimine and peiminine within F-F. CONCLUSION This study employs UHPLC-Q-TOF-MS technology to conduct an exhaustive analysis of the chemical composition of F-F. From the results, it can be seen that the F-F possess potential medicinal value. This study provides scientific basis for the rational development and sustainable utilization of its resources.

OBJECTIVE To evaluate the difference of pharmacokinetics and tissue distribution of folate-modified cantharidin/baicalin liposomes(FA-Can&Bai@Lips) in normal and tumor-bearing rats. METHODS HepG2 cell suspension was inoculated into the right armpit of rat to induce tumor bearing animal models. After tail vein injection of FA-Can&Bai@Lips, plasma and tissues(heart, liver, spleen, lung and kidney) were collected at different time points. The concentrations of cantharidin and baicalin in plasma and tissues were determined by ultra high performance liquid chromatography-mass spectrometry(UPLC-MS/MS). The pharmacokinetic parameters were calculated by Phoenix WinNonlin software, and the pharmacokinetic and tissue distribution differences of FA-Can&Bai@Lips in normal and tumor-bearing rats were analyzed and compared. RESULTS The UPLC-MS/MS method for simultaneous determination of cantharidin and baicalin has a good linear relationship. The specificity, accuracy and precision, extraction recovery rate and matrix effect, residual effect and stability meet the requirements of biological sample detection. The pharmacokinetic parameters showed that the area under the curve(AUC_0-t,AUC_0-∞), the mean residence time(MRT_0-t, MRT_0-∞) of cantharidin and baicalin in plasma of tumor-bearing rats were significantly higher than those in normal rats(P<0.05), which increased by 55.68%, 72.50%, 43.10%, 45.95% and 15.10%, 42.54%, 9.09%, 10.53% respectively. The half-life(t_1/2) of cantharidin was slightly shortened in tumor-bearing rats with no significant difference, while the t_1/2 of baicalin was significantly prolonged(P<0.05). The apparent volume of distribution(Vd) and clearance(CL) of cantharidin and BA were significantly decreased. The results of tissue distribution showed that the concentrations of cantharidin and BA in the liver of tumor-bearing rats were significantly higher than those of normal rats, and the other tissues were significantly lower(P<0.01). CONCLUSION The UPLC-MS/MS method is rapid, simple, specific and sensitive, and can be used for the determination of cantharidin and baicalin in plasma and tissues of rats. It is confirmed that the tumor-bearing status can significantly affect the pharmacokinetics and tissue distribution of FA-Can&Bai@Lips in vivo.

OBJECTIVE To explore the effect of low molecular weight heparin sodium on regulatory T cell(Treg) differentiation in rats with unexplained recurrent pregnancy loss(URPL) by regulating the miR-33a-3p, sphingosine-1-phosphate receptor 1(S1PR1) expression. METHODS Sixty female rats were randomly divided into blank control group(Control), negative control group(NC), URPL group, low molecular weight heparin sodium group(LMWH), and fingolimod(FTY720) group, with 12 rats in each group. On the 8th and 12st day of pregnancy, subcutaneous injection of anticardiolipin antibodies(ACA)-IgG into multiple parts of the back was used to induce rat URPL model. LMWH group and FTY720 group were administered on the basis of URPL group. On the 0th and 15th day of pregnancy, LMWH group rats were subcutaneously injected with low molecular weight heparin sodium(420 IU·kg^-1), while FTY720 group rats were injected with FTY720(100 μg·kg^-1) via tail vein, control group, NC group and URPL group rats were injected with an equal amount of physiological saline via the tail vein. After treatment, weigh the embryo and calculate the embryo absorption rate. HE staining was used to observe the pathology of placental tissue. Enzyme linked immunosorbent assay(ELISA) was used to detect interleukin(IL)-10 and transforming growth factor-β1(TGF-β1) levels in serum. Real time quantitative polymerase chain reaction(RT-qPCR) and Western blot were used to detect miR-33a-3p, S1PR1, forkheadbox protein 3(FOXP3), cytotoxic T lymphocyte associated protein 4(CTLA-4), and glucocorticoid induced tumor necrosis factor receptor(GITR) mRNA and protein levels in placental tissue. Flow cytometry was used to detect the number of Treg cells in placental tissue. RESULTS The placental cells of rats in Control group and NC group were arranged neatly, and structure was clear. Compared with Control group, There was a large amount of inflammatory cell infiltration, cell proliferation, and edema in placental tissue in URPL group, embryo quality was reduced, embryo absorption rate was increased, IL-10 and TGF-β1 levels in serum were decreased, miR-33a-3p, FOXP3, CTLA-4, GITR mRNA and protein levels in placental tissue were decreased, S1PR1 mRNA and protein levels was increased, the number of Treg cells was decreased(P<0.05). Compared with URPL group, the pathological damage to the placental tissue in LMWH group and FTY720 group were significantly reduced, with a small amount of inflammatory cell infiltration and edema visible, embryo quality was increased, embryo absorption rate was decreased, IL-10 and TGF-β1 levels in serum were increased, miR-33a-3p, FOXP3, CTLA-4, GITR mRNA and protein levels in placental tissue were increased, S1PR1 mRNA and protein levels was decreased, the number of Treg cells was increased(P<0.05). There was no statistically significant difference in the above indicators between LMWH group and FTY720 group(P>0.05). CONCLUSION Low molecular weight heparin sodium maybe promote Treg cells differentiation in URPL rats by regulating the miR-33a-3p, S1PR1 expression.

OBJECTIVE To synthesize and prepare nitric oxide donor(NODonor)-silicon phthalocyanine(SiPc) conjugated prodrug self-assembled nanoparticles(NO-SiPc-NO@NPs) and preliminarily evaluatetheir formulating properties and pharmacodynamics. METHODS Furoxan NO donor-phthalocyanine silicon photosensitizer couplings were synthesized by chemical reactions. The NO-SiPc-NO@NPs were prepared using a nanoprecipitation method,and the effects of different concentrations of NO-SiPc-NO, rotational speed, the volume ratio of the organic phase to the aqueous phase, and the content of the stabilizer DSPE-PEG_2K on particle sizes, polydispersity index(PDI), and Zeta potential of NO-SiPc-NO@NPs were investigated to obtain the better prescriptions. The release of NO and reactive oxygen species(ROS) yields as well as the photostability of NO-SiPc-NO were detected by the Griess method and the chemical probe method, respectively. On this basis, the storage stability and in vitro release of NO-SiPc-NO@NPs in phosphate buffer salt solutions of different pH were investigated. Finally, the photodynamic effect of nanoparticles was detected by CCK-8 method, and the effect of self-assembled nanoparticles on intracellular NO was observed by fluorescent probe. RESULTS The ¹H-NMR results showed that NO-SiPc-NO was successfully synthesized. The optimal preparation process conditions were:NO-SiPc-NO concentration of 2.5 mg·mL^-1, rotational speed of 1 000 r·min^-1, organic phase to aqueous phase volume ratio of 1∶3 and stabilizer content of 40%. The prepared self-assembled nanoparticles were(111.467±3.365) nm, (0.123±0.035) and (-11.433±0.850) mV in particle size, PDI and Zeta potential, respectively. The nanoparticles in transmission electron microscopy were spherical or spheroidal in shape, with a more intact morphology and homogeneous distribution. The results of NO and ROS release showed that the nanoparticles could release NO and ROS in solution with good photostability. NO-SiPc-NO@NPs were stable under both conditions, and there was no significant change in particle size, potential, PDI, encapsulation rate and drug loading. The results of in vitro drug release experiments showed that NO-SiPc-NO@NPs had a slow release and that their release followed a one-level kinetic model. CCK-8 experiments showed that all nanoparticle groups showed dose-dependent cytotoxicity, and the light-exposed NO-SiPc-NO@NPs had a stronger photodynamic effect on MCF-7 cells. In NO assay experiments, NO-SiPc-NO@NPs can produce large amounts of NO intracellularly. CONCLUSION NO-SiPc-NO@NPs are successfully prepared and the prepared self-assembled nanoparticles have good photodynamic activity and realize effective delivery of NO, which lays a theoretical foundation for the synergistic treatment of gas therapy and photodynamic therapy.

OBJECTIVE To study the in vitro transdermal characteristics of Liuwei Dihuang gel and evaluate the safety of skin medication. METHODS The improved vertical Franz diffusion cell method was used for in vitro transdermal experimental and the skin of rat was used as the permeation barrier. The cumulative permeation amount and rate of morroniside, loganin, and paeonol in the receiving medium were determined by HPLC, to investigate the in vitro transdermal characteristics of Liuwei Dihuang gel. The skin irritation was observed after single or multiple application of Liuwei Dihuang gel on intact and damaged skin of guinea pig. RESULTS The cumulative permeation amount of morroniside, loganin and paeonol within 12 h were 246.56, 298.47 and 369.89 μg·cm^-2, and the cumulative permeation rate were 19.67%, 21.55% and 39.18%, respectively. Liuwei Dihuang gel is not irritating to both intact and damaged skin of guinea pigs. CONCLUSION The in vitro transdermal performance of Liuwei Dihuang gel is good. The in vitro transdermal process conforms to the Higuchi kinetic equation. It has no irritation effect on the skin of guinea pigs. It is safe and reliable for skin external use and has good linical application potential.

OBJECTIVE To establish an efficient method for quantification of adalimumab in human plasma based on UPLC-Q Exactive-Orbitrap MS platform combined with immuno-affinity enrichment strategy. METHODS Candidate surrogate peptides were screened by full MS/ddMS² and the selective surrogate peptide was quantitatively analyzed by parallel reaction monitoring. Immunoglobulins and therapeutic antibodies in human plasma were extracted by magnetic beads coupled with protein A. The proteins were denatured at high temperature and digested by trypsin. Stable-isotope labeled adalimumab was used as internal standard. RESULTS The peptide GLEWVSAITWNSGHIDYADSVEGR in variable region of adalimumab heavy chain was selected as signature peptide, showing specificity and selectivity. Adalimumab demonstrated good correlation within the range of 1-32 μg·mL^-1. Precision, accuracy and total error all met the verification requirements. Thirty-one clinical samples had measurable adalimumab concentrations by the established method and ELISA method, yielding a good correlation. The mean of difference between the two methods was 0.5 μg·mL^-1. CONCLUSION The universal immuno-affinity mass spectrometry method established in the study is suitable for quantification of therapeutic antibodies, and can accurately and precisely determine adalimumab concentration, which provides a strategy for clinical monitoring.

OBJECTIVE To establish the HPLC fingerprint and multi-component quantitative determination method of Glehniae Radix, and study the dispersion in quality of Glehniae Radix of different regions and residual root bark ratio by chemical pattern recognition. METHODS HPLC method was used to establish the fingerprint and determine the contents of seven active components. The chromatographic column was ShimNex CS C₁₈(4.6 mm×250 mm, 5 μm), the mobile phase was methanol-0.4% phosphoric acid water, gradient elution was conducted at a volume flow rate of 1.0 mL·min^-1, the column temperature was maintained at 30 ℃, and the detection wavelength was set at 204 and 248 nm. Fingerprint similarity was used to evaluate different regions of Glehniae Radix. Cluster analysis, principal component analysis and discriminant analysis were used to analyze the quality and the differential components of Glehniae Radix of different residual root bark ratio. RESULTS A total of 18 common peaks were found in the fingerprints of 60 batches of Glehniae Radix fingerprints from different regions, and seven peaks were identified, including facarindiol, panaxynol, xanthotoxin, bergapten, imperatorin, isocorallol and isoimperatorin. The similarity of 60 batches of samples was 0.814-0.996. The multi-component contents of the 190 batches of Glehniae Radix were divided into two groups by cluster analysis, and the samples with residual root bark ratio ≤1/3 and >1/3 were significantly distinguished. The results of PCA and HCA were consistent. The different quality markers were panaxynol and facalindiol. CONCLUSION The established HPLC fingerprint and multi-component content determination method are simple, stable, and reliable, which can provide a basis for the quality control, overall evaluation and standardized processing methods of Glehniae Radix.

OBJECTIVE To establish a high resolution mass spectrometry method for characterization of recombinant human follicle-stimulating hormone disulfide bonds. METHODS For pre-treatment, follicle-stimulating hormone(FSH) was diluted to 0.5 mg·mL^-1 with diluent, subtilin, trypsin, and PNGase F were added into the stock solution respectively, and 1 μL 10% formic acid was finally added to stop the reaction. For LC-MS analysis, ACQUITY UPLC Peptide BEH C₁₈(2.1 mm×100 mm, 1.7 μm) column was used, the mobile phase was 0.1% formic acid/aqueous solution(A)-0.1% formic acid/acetonitrile solution(B), and gradient elution was performed. The mass spectrum acquisition mode was ddMS2, the ion source was ESI⁺, and the scanning range was m/z 250-2 000. RESULTS The 6-p-disulfide bond of β-subunit was successfully identified, with the values β as followes: C3=β:C51. β:C17=β:C66; β:C20=β:C104; β:C28=β:C82; β:C32=β:C84; β:C87=β:C94. The 5-p-disulfide bond of α-subunit was successfully identified, α:C7=α:C31; α:C10=α:C60; α:C28=α:C82; α:C32=α:C84; α:C59=α:C87. CONCLUSION The localization method of disulfide bond of recombinant human follicle stimulating hormone(hFSH) is established, which provides a new idea for the quality control of disulfide bond connection.

OBJECTIVE To establish and implement the drug delivery service mode as part of internet healthcare in hospitals, and evaluate its practical effectiveness,in order to grand out the last mile of the internet patient service and provide valuable experience and reference for better serving patients. METHODS Based on the requirements of relevant national laws and regulations and the actual situation of our hospital, the drug delivery process was designed, and the distributable drug list was established. The drug delivery service was carried out by making full use of information technology and logistics system through comprehensive and detailed system construction and process design. Drug orders issued until April 30,2024 were collected and the economic and social benefits generated by the delivery of drug were analyzed. RESULTS A safe and controllable drug delivery service mode was established as part of the internet healthcare with the characteristics of hospitals. By April 30, 2024, 94 370 drug orders had been distributed, covering 31 provinces and cities in China. Taking the closest patients from Hebei province as an example, each patient can save 660 yuan on transportation and accommodation expenses per visit, patients in this province have saved about a total of 15 million yuan in expenses. CONCLUSION The development of internet-based diagnosis and treatment effectively solves the problem of patient follow-up visits, and drug delivery is a key link in the process of internet healthcare. Sending drugs to the hands of patients not only saves the patient's economic cost and time cost, but also ensures the continuity of patient treatment during epidemics such as COVID-19 and H1N1. With the expansion of the scope of online medical insurance settlement, this model will provide efficient and convenient services to more patients.

OBJECTIVE To explore investigator-initiated trial(IIT) approval management with a focus on risk prevention and control in the absence of guidelines for IIT approval management. METHODS Through a combination of literature review and practical experiences, this study analyzed the challenges encountered in the approval process and explored its potential risks, outlined the approval process following the promulgation of the management regulations, summarized its key aspects, and assessed the significance of the approval process with a focus on risk prevention and control.RESULTS There exist inherent risks in areas such as research design, participant protection, research team dynamics, and research funding. The authors advocate for the implementation of compliance review, peer expert evaluation, and a hierarchical approval review by expert technical committee, emphasizing scrutiny of scientific validity, feasibility, and funding adequacy. CONCLUSION The proactive identification and prevention of potential risks are highlighted as crucial benefits of the approval management. Centralized management of IIT projects, development of data management platforms, and diversification of funding channels to enhance risk mitigation should be further explored.