Fc region of a monoclonal antibody usually contains two N-glycosylation, which have a profound influence on its structure and function. Here, we review the relationship between various glycoforms and their impact on structure and function of monoclonal antibodies, along with the technologies for glycosylation analysis. In addition, some related Chemistry, Manufacturing and Controls (CMC) regulatory considerations are also discussed, such as specification improvement, biosimilarity assessment as well as comparability of pre- and post-process changes in glycosylation.

Fifteen 9-substituted palmatine (1) derivatives were synthesized and evaluated for their anti-Helicobacter pylori (Hp) activities in vitro. Structure-activity relationship studies revealed that introducing appropriate substituted secondary amino group at position 9 of lead 1 might be beneficial for potency. Among them, compound 5a showed the most potential activity against metronidazole (Met) resistant Hp isolates with minimal inhibitory concentrations (MICs) of 4 μg·mL^-1, much better than that of lead 1. Compound 5a displayed satisfactory safety profile in acute toxicity assay. Molecular docking suggested that 5a might act on Hp urease. The results provided key scientific evidence for the development of 1 derivatives into a new class of anti-Hp component.

Intestinal Peyer's patches (PPs) are important sites to induce mucosal immunity response. As a kind of specialized epithelial cells in PPs, microfold cells (M cells) have a unique ability to take up antigens and can promote systemic immune response by transferring antigens to dendritic cells (DCs) in PPs. Selecting a suitable drug delivery vehicle and modifying the vehicle with a specific ligand can target drugs and bioactive substances to M cells to exert a therapeutic effect on intestinal immune-related diseases. This paper reviews the relevant literatures in the past 20 years, and summarizes and analyzes the ligands, types of vehicles, material properties that target M cells and main factors affecting the uptake of M cells, in order to provide construction ideas and experimental methods that can be worthy of reference for the study of PPs M cells-targeting drug delivery strategies.

Cyclophosphamide (CPA) is the first-line chemotherapy for many tumors, but its overdose will lead to hepatotoxicity. This study aims to investigate whether the combined administration of oxymatrine (OMT) with CPA will aggravate the hepatotoxicity induced by CPA and its engaged mechanism. The expression of hepatic Cyp2b10 mRNA and CYP2B10 protein was detected by qPCR and Western blot in mice at different times after OMT (100 mg·kg^-1) administration. Mice were given with different doses of OMT (intragastric administration, ig) every day. At the same time, CPA (200 mg·kg^-1) was also intraperitoneally injected into mice every other day. After 10 days, serum alanine/aspartate aminotransferase (ALT/AST) activity, the mortality of mice and hepatic mRNA expression of Cyp2b10 were detected. Furthermore, the correlation among ALT/AST activity, the mortality and Cyp2b10 mRNA expression was analyzed. All animals were received humane care according to the institutional animal care guidelines approved by the Experimental Animal Ethical Committee of Shanghai University of Traditional Chinese Medicine. The results showed that OMT itself enhanced hepatic mRNA and protein expression of Cyp2b10 (P < 0.05), and increased liver enzymatic activity of CYP2B10 in mice (P < 0.05). In mice treated with CPA plus OMT, OMT obviously enhanced the mortality of mice induced by CPA (from 33.3% to 58.3%). The results of serum biochemical analysis and hepatic mRNA expression of Cyp2b10 showed that OMT further enhanced the increased serum ALT/AST activity and hepatic Cyp2b10 mRNA expression in mice (P < 0.05). There was a good correlation between serum ALT/AST activity and mortality or hepatic Cyp2b10 mRNA expression. These results showed that OMT could enhance hepatic Cyp2b10 mRNA expression and increase liver CYP2B10 enzymatic activity, and then promoted the metabolism of CPA, and thus aggravated CPA-induced hepatotoxicity in mice.

In this study, we used molecular docking technology and validation experiments in vitro to screen the active ingredients of Erzhi pill for treating osteoporosis. Firstly, the compound in Erzhi pill was docked with ten osteoporosis-related targets by molecular docking technology, and the five active compounds, salidroside, specnuezhenide, tyrosol, quercetin, and wedelolactone, were initially selected. Then, MC3T3-E1 osteoblasts were cultured in α-Mem medium containing different concentrations of compounds, and the cell proliferation rate and mineralized nodules were tested. The verification results showed that the proliferation of MC3T3-E1 cells with salidroside, specnuezhenide and quercetin were more obvious, and salidroside has a better effect on the mineralization of MC3T3-E1 cells than those of specnuezhenide and tyrosol. The molecular docking technology coupled with validation experiment in vitro can be used for the screening of anti-osteoporotic active ingredients of Erzhi pill, and it can also provide a method for the study of effective ingredients of traditional Chinese medicine.

Delivering water-soluble drugs via carriers often causes problems such as low loading and rapid releasing, so it is an urgent need to construct a high-load sustained-release drug delivery system for the clinical application of water-soluble drugs. Two-dimensional layered nanomaterials exhibit great potential in drug delivery due to their high specific surface area. In this study, bulk graphitic carbon nitride (b-g-C₃N₄) was obtained by calcination of urea. Graphitic carbon nitride nanosheets (g-C₃N₄-NS) were made from an alkali chemical-ultrasonic-assisted stripping process. Scanning electron microscopy, transmission electron microscopy and atomic force microscopy were adopted to observe the morphological characteristics of g-C₃N₄-NS, while the structural characteristics of g-C₃N₄-NS were analyzed by X-ray diffractometer and Fourier transform infrared spectroscopy. Ultraviolet spectrometry and fluorescence spectrometry were used to investigate the optical properties of g-C₃N₄-NS, and scanning electron microscopy and X-ray diffractometer were employed to investigate the stability of g-C₃N₄-NS. Polyethyleneimine (PEI) was applied in the study to functionally modify g-C₃N₄-NS, and salvianolic acid B (Sal B) was used as a water-soluble drug model to investigate the loading capacity and drug releasing behavior of g-C₃N₄-NS. The results showed that g-C₃N₄-NS had a sheet structure, and it is easy to self-assemble in layers in the ionic environment to create flocculating settling. PEI modification can lead to the switching in the surface charge of g-C₃N₄-NS and significantly improve its stability. The results of cytotoxicity test and zebrafish embryo toxicity test showed that the toxicity was low when the concentration of PEI-g-C₃N₄-NS was less than 800 μg·mL^-1. The large specific surface area and surface charge of PEI-g-C₃N₄-NS allow the maximum load factor over Sal B to reach 327.4%. In addition, PEI-g-C₃N₄-NS can continuously release drugs slowly, with a cumulative release rate of 79.2% in seven straight days. The release process conforms to the Higuchi equation. In summary, g-C₃N₄-NS modified by PEI exhibits good biocompatibility and high stability, and shows great potential in high-load and sustained-release applications of water-soluble drugs.

Degradation of dexamethasone (DXM) is inevitable in the release test of dexamethasone implants (DI). In the release test conducted with flow-through cell method, the measured release curves of DI started to fall when cumulative release reached 70%-80%. Studies have shown that DI demonstrates a zero-order release rate of drug within every sampling interval, and a zero-order rate degradation in water (containing 0.05 mg·mL^-1 benzalkonium chloride). Hence, this study establishes a double zero-order model (DZOM) to calculate the release during sampling intervals with the formula R_i = [R_im-R_(i-1)m×(C_in/C_i0)]×2/(1+C_in/C_i0). At each sampling interval, we measure the initial and final drug contents in the release medium, and the concentrations of the active pharmaceutical ingredient (API) in the release medium obtained at the same condition of release test, to calculate the total released DXM from the implants including the degraded drug. This paper has also analyzed the reasons for the fluctuations in the drug release curve and the errors in the DZOM and provided solutions. Experimental results show that the DZOM has effectively solved the problems encountered in the normal release method (NRM). The DZOM can be a potential solution to drug degradation problems in the release tests of long-acting injections.

Asparagus cochinchinensis is a commonly used traditional Chinese medicine with steroidal saponins as its main active ingredients. Due to the structural similarity and size of the steroidal saponins, these compounds cannot always be effectively separated by a combination of normal phase silica gel column chromatography and reversed phase ODS column chromatography. In this experiment, chromatographic columns with different separation mechanisms were systematically screened, and it was found that a chiral chromatographic cellulose column could effectively separate these components. This column was used to separate 3 mixtures to obtain 6 single compounds (1-6). Structural identification showed that the singular structural difference between these poorly separated components resides in a terminal glycosyl group (xylose or rhamnose) in the C-3 glycosyl chain, and compounds 4 and 6 are two new steroidal saponins. Since the structures of compounds are often unknown during the isolation and purification of natural products, chiral columns are rarely used. This study suggests that chiral chromatographic columns are a valuable option for natural products that are difficult to separate by conventional means.

In recent years, cancer immunotherapy has become an important field of basic and applied researches of cancer immunology. Cancer immunotherapy mainly includes cancer vaccine, oncolytic virus therapy, chimeric antigen receptor T cells (CAR-T cells), immune checkpoint blocks, monoclonal antibodies, and other strategies. Among them, monoclonal antibody-based cancer immunotherapy has the fastest development. In the past 20 years, monoclonal antibody has become one of the drugs with remarkable curative effect and novel type for human malignant tumors, especially the monoclonal antibody targeting immune checkpoints, play an important role in immunotherapy. In this review, we will summarize the current situation of monoclonal antibody-based cancer immunotherapy, potential immune modulatory mechanism, antibody targeting molecules and its immunotherapeutic agents, and explore the trend of monoclonal antibody-based agents in cancer immunotherapy.

This study aimed to establish a method for identification of the prescription components of Pule'an Tablet based on DNA barcoding technology. Sixteen samples have been collected from 8 different companies, and their DNA were purified using Plant and Animal Genomic Kits. The amplification rates of ITS2 were both 100%, and the amplification rates of COI were 43.75% and 56.25% for these samples' DNA purified using plant and animal kits, respectively. For ITS2, 12 samples obtained high-quality sequencing traces and then were identified as containing Brassica campestris. The other 4 samples showed crucial SNP peaks in the sequencing traces, which were assigned to be B. campestris and B. nigra on the basis of cloning and sequencing experiments. For the PCR products of 9 samples from COI universal primers, two samples were directly sequenced and identified as aphids, and 7 other samples were subjected to cloning and sequencing experiments. Finally, we obtained 82 clone sequences and found that Apis mellifera was detected only in 5 of the remaining 7 samples, and pathogens or pests were detected in all these 7 samples. To solve the failure of bee source detection caused by exogenous contaminations based on COI universal primers, we designed two new COI primer pairs of Apis genus. The amplification rates of both primer pairs were 43.75%, and they were identified as A. mellifera. A total of 9 bee source-free Pule'an Tablet samples from 3 different batches were produced by the same company, and each batch contained 3 replicates. Thus, we speculated that raw rape pollen materials for these 9 samples was not collected by bees. This study proposes an identification method for the prescription components of Pule'an Tablets based on ITS2 and COI sequences, which will provide scientific basis and technical guidance for quality control and market regulation of Pule'an Tablets.