OBJECTIVE To develop the methods for biological activity assay and immunological characteristics analysis of anti-SARS-CoV-2 neutralizing antibodies. METHODS The binding affinity of 9MW3311 Fab and S1-RBD were analyzed by biolayer interferometry. Enzyme linked immunosorbent assay (ELSA) and fluorescence activated cell sorter (FACS) were used to evaluate the relative binding activity to S protein and blocking activity to angiotensin converting enzyme 2(ACE2) of 9MW3311 antigenbinding fragments (Fab). In vitro cytological activity of neutralizing antibody was evaluated by pseudovirus system. The binding affinities of neutralizing antibody Fc with Fc receptor (Fcγ) and Fc receptor neonatal (FcRn) receptor were determined by surface plasmon resonance (SPR). The binding activity of Fc and complement component 1 (C1q) receptor was determined by ELISA. The antibody-dependent cell-mediated cytotoxicity(ADCC) and complement dependent cytotoxicity(CDC) of neutralizing antibodies were determined by peripheral blood mononuclear cell (PBMC) method. The antibody-dependent enhancement (ADE) effect was evaluated using pseudovirus system. RESULTS The affinity constants (KD) of 9MW3311 and reference to S1-RBD were 1.15×10^-9, 1.01×10^-9, 1.15×10^-9 and 9.45×10^-10, respectively. ELISA and FACS showed that the binding activities of neutralizing antibodies were 101%, 96%, 100% and 98%, 113%, 108%, respectively. ELISA and FACS showed that the blocking activities of neutralizing antibodies against ACE2 were 100%, 95%, 91% and 94%, 101%, 94%, respectively. The neutralizing activities of the three batches of neutralizing antibodies against pseudovirus were 91%, 93% and 108%, respectively. The three batches of 9MW3311 and reference showed the same affinity constants (KD) with Fcγ and FcRn. 9MW3311 showed no ADCC and CDC activity. L234A/L235A (LALA) mutant of 9MW3311 could effectively avoid ADE effect. CONCLUSION The methods for analysis of the biological activity and immunological characteristics of anti-SARS-CoV-2 neutralizing antibodies are preliminarily established and can be used for routine quality control and release.

OBJECTIVE To study the components of Rhizoma Anemarrhenae-Cortex Phellodendri Chinesis (RA-PC) herb-pair absorbed into blood in normal rabbits. METHODS Rabbits were fed with the decoction of RA-PC herb pair (4.5 g·kg^-1) once a day for 7 days, then blood samples were taken. High-performance liquid chromatography-quadrupole-electrostatic field orbital trap (UPLC-Q-Exactive Orbitrap-MS) technique was applied for the analysis, and fragments were split in both positive and negative ion modes. The compositions into blood were determined according to retention time, accurate relative molecular mass, and secondary mass spectra. RESULTS A total of 54 components were identified in the drug-containing serum of rabbits, including 46 prototype components, which were alkaloids, flavonoids, saponins, glycosides, phenolic acids, phenylpropanoids, lactones, and phenolic derivatives. The nine metabolites were derived from berberine, jatrorrhizine, berberrubine, and mangiferin, respectively. CONCLUSION RA-PC contains 46 chemical components that can enter blood of normal rabbits, and may also contains four chemical components which are metabolized into nine metabolites.

OBJECTIVE To establish a nano sustained release drug delivery system polysuccinimide(PSI) and hydroxyapatite(HAP) with a hard core-soft coating structure and study the delivery of anti-tumor drug regorafenib. METHODS HAP and PSI were synthesize in the article. The optimal formulation and preparation process of the regorafenib-PSI-HAP was determined by single factor analysis and the box-behnken design(BBD) response surface method. The encapsulation efficiency, drug loading captivity, drug release in vitro, particle size and distribution, Zeta potential and SEM images of regorafenib-PSI-HAP were examined. RESULTS The UV spectrophotometry results showed that the maximum absorption wave of the regorafenib was at 264 nm, the stand curve was A=94.461ρ+0.0304, r²=0.999 8. Meanwhile, between the 2-20 μg·mL^-1 of the regorafenib solutions possessed a good linear relationship at this wave. The optimal formulation prepared using MEM detected by BBD was that the concentration of regorafenib was 7.28 mg·mL^-1, the mass ratio of HAP to regorafenib was 0.63, and the mass ratio of PSI to HAP was 3.93. The in vitro drug release test showed that PSI-HAP exhibited pH-sensitive drug release, the regorafenib-PSI-HAP can be released totally above pH 7 solution. The PSD was around 300 nm, and Zeta potential was between -10 and -17 mV which was similar to the human cell membrane. CONCLUSION The proposed drug delivery system can be integrated into non-agglomerated spherical nanoparticles of uniform size through a facile preparation process. The particles are easy to sterilize, and large-scale production may be achieved easily. The drug release of PSI-HAP is pH sensitive and stable. Both PSI and nano HAP possess the effect of anti-tumor cell proliferation which is very suitable for the delivery of anti-tumor drugs such as regorafenib in vivo.

OBJECTIVE To explore an evaluation method of sterile operation in pharmacy intravenous admixture services(PIVAS). OBJECTIVE The US intravenous quality assurance(IVQA) media fill test kit was used to verify the aseptic operation of the mixed dispensing personnel in our hospital, and the feasibility of applying the media fill test in the PIVAS was preliminarily explored. RESULTS The media fill test was easy to operate without harsh culture conditions and testing equipment. The test results were different under two different mixing conditions, which can verify the operation of personnel. CONCLUSION The media fill test is applicable to the quality control of personnel in sterile dispensing operations.

OBJECTIVE To verify the accuracy of the “water content-conductivity fitting curve” model for determining the critical point of microemulsion O/W phase, and to further use this model to optimize the microemulsion of tanshinone extract. OBJECTIVE The “water content-conductivity fitting curve” model was used to determine the phase critical point of microemulsion into O/W type. According to the proportion of microemulsion oil phase, total surfactant and water phase, the pseudo ternary was drawn using Origin 2021 software. The phase diagram was used to screen out the proportions of the larger milk-forming areas; the average particle size and PDI of the microemulsion were used as the inspection indicators, and the star point design-response surface method was used to optimize the microemulsion; high-speed centrifugation and standing at 4 ℃ and 25 ℃ for 60 days, to examine the stability of the microemulsion. RESULTS The optimized microemulsion formulation was(18.11% medicated oil phase, 61.21% Tween 80, 20.68% 1,2-propanediol, the water critical point is 64.29%), K_m was 2.96, preparation temperature was 27.4 ℃, under different experimental conditions, the stability of the microemulsion was good. CONCLUSION The O/W phase critical point determined by the “water content-conductivity fitting curve” model has high accuracy, and the optimized microemulsion has good stability, which achieves the expected research goals.

OBJECTIVE To establish a method for the determination of 32 elements in COVID-19 vaccine. OBJECTIVE The 32 elements were determined by inductively coupled plasma optical emission spectrometer(ICP-OES) through semi-quantitative screening and quantitative detection, and the specificity, linearity, limit of quantification, detection limit and recovery rate of the ICP-OES method were investigated to evaluate the feasibility of the method for the detection of trace elements in vaccines. RESULTS The linear relationship of each element was good within the detection mass concentration range, and the recoveries of element added were 94.06%-115.50%. When applied to the detection of the samples at each accelerated placement point, except for the trace contents of B and Si elements, which did not increase significantly after long-term placement, the contents of other elements were far less than the prescribed limits. CONCLUSION The ICP-OES method is established. And the method has good specificity, reliable results, simple and rapid, and is suitable for the determination of 32 elements in COVID-19 vaccine.

OBJECTIVE To screen one Monascus purpureus stain that has high yield of lovastatin and provide potential strains for the development of red yeast rice(RYR) industry. OBJECTIVE Twelve RYR samples were collected, and Monascus sp. was isolated from these samples using traditional isolation technology and identified by morphology and ITS sequencing. All M. purpureus strains were screened and their total lovastatin contents [including lactone lovastatin(C₂₄H₃₆O₅) and acid lovastatin(C₂₄H₃₈O₆)] were determined by high performance liquid chromatography(HPLC). Soft SPSS 25.0 was used for variance analysis. RESULTS A total of 35 Monascus sp. strains were isolated and 23 of them were identified at specie level, including 11 M. purpureus strains, 10 M. ruber strains and 2 M. aurantiacus strains. There was significant difference(P<0.05) in lovastatin production among 11 M. purpureus strains. Among them, SM-1 isolate produced the highest amount of total lovastatin, with the value reaching (4.652±0.001) mg·g^-1, followed by M4-1, F1-3, and F5-3 strains with the value reaching (2.903±0.033) and (3.108±0.009) mg·g^-1, respectively. M4-5 isolate produced the lowest amount of lovastatin with the value of (2.712±0.014) mg·g^-1. CONCLUSION Fungal strain is chaotic in the fermentation of RYR on the market. A high-yielding lovastatin strain SM-1 is screened and can be used a potential strain for RYR production.

OBJECTIVE To build a text-convolutional neural network(Text-CNN)-based prediction model for anti-Staphylococcus aureus(S.aureus) activity and identify anti-S.aureus hits by virtual screening. OBJECTIVE A dataset containing 26327 compounds annotated with S.aureus activity data was collected and curated from the ChEMBL database. Ten pairs of training and test sets were generated by random partition for 10 times and then 10 models were built using the Text-CNN algorithm. The best-performing model was determined by model evaluation and further studied by Y-randomization test and applicability domain analysis. Following that, the best-performing model was used to virtually screen the in-house chemical library, by which the potential antibacterial agents were determined. The micro-broth dilution method was used to test anti-S.aureus activity of the potential hits. RESULTS The machine-learning model(named Text-CNN3) performed well in classification. Evaluated on the test set, its Mathews correlation coefficient was 0.573 and the area under the ROC curve was 0.881. With this model for virtual screening as well as antibacterial screening, compounds Y5 and Y7 were identified as antibacterial compounds, with minimum inhibitory concentrations(MIC) of 8 and 4 μg·mL^-1, respectively. CONCLUSION The Text-CNN3 model in this study is effective to identify anti-S.aureus compounds, while the antibacterial hits Y5 and Y7 are worthy of further study.

OBJECTIVE To design and synthesize N-substituted methylene-3-(4-(pyridin-3-yl)pyrimidin-2-yl)amino)benzohydrazide derivatives, and evaluate their anticancer activities. OBJECTIVE Using 3-aminobenzoic acid ethyl ester as the starting reagent, a series of target compounds 4a-4p were synthesized in a 4-step reaction, and all of the target compounds were structurally characterized by ¹H-NMR, ¹³C-NMR and HRMS. MTT assay was used to detect the inhibitory effect of synthetic compounds on breast cancer cells. RESULTS It's showed that the target compounds had certain inhibitory activity on the growth of breast cancer cells, among which compound 4e was the best one, showing obvious inhibitory effect on breast cancer cells. The IC₅₀ value of breast cancer cell line MCF-7 was (2.30±0.34)μmol·L^-1, and the toxicity of 4e to normal mammary epithelial cells was low(IC₅₀>100 μmol·L^-1). Thus, 4e is better than the positive drug cisplatin. In addition, 4e could significantly up-regulate the protein expression level of Nur77 and induce the S-phase arrest of MCF-7. CONCLUSION This series of compounds have good anti-cancer activity and are of interest for further study.

OBJECTIVE To analyze the quality differences of Cyathulae Radix from different manufacturers on the market based on national drug sampling and testing in 2022, then analyze the problems and put forward suggestions to provide basis for the quality supervision and standard improvement of Cyathulae Radix. OBJECTIVE Inspection was conducted according to corresponding standards, inspection reports were issued, and then quality evaluation and analysis were conducted. At the same time, problems were found and suggestions were made based on targeted exploratory research on quality problems, standard problems and other related problems found in the inspection. RESULTS A total of 334 batches of Cyathulae Radix samples were taken. According to the standard inspection, two batches of samples were found to be substandard in character, and Cyathulae capitatae Radix was used as Cyathulae Radix. At the same time, it was found that the content determination methods in the standard were too complex, and the specified limit for content determination was too low. In the exploratory study, the unique component achybidensaponin I was detected in 27 batches of Cyathulae Radix and 7 batches exceeded the specified limit, which was mixed with Achyranthes bidentata. Plant growth regulator (paclobutrazol) was detected in 270 batches of Cyathulae capitatae Radix. The detection of heavy metals and harmful elements cadmium(Cd) in 114 batches exceeded the limit. Finally, the quality standard of Cyathulae Radix was improved, the supplementary test method was established, and the risk warning was put forward. CONCLUSION Through the national sampling inspection, the relevant problems in the prepared slices and quality standards of Cyathulae Radix are found, suggesting that the quality standards should be improved and the quality supervision of Cyathulae Radix should be strengthened.