This study aims to synthesize fluorinated hyperbranched poly(amido amine)s for the delivery of influenza DNA vaccine. Hyperbranched poly(amido amine)s (HP) were synthesized by using Michael-type polyaddition and then fluorinated to obtain fluorinated polymers (F-HP). The target gene was amplified by designing specific primers to construct the eukaryotic expression plasmid of influenza viral PR8 nucleoprotein (NP) gene as DNA vaccines. Then the polyplexes (F-HP/NP) were prepared by the electrostatic interactions between polymers and plasmid. The results suggested that the molecular weight of HP was 59.7 kDa and polydispersity index (PDI) was 2.67. The fluorine content in F-HP was found to be 20% (w/w). Transmission electron microscopy (TEM) revealed that the polyplexes had spherical shapes with sizes around 100 nm (w/w 5-10) and decreased with increasing w/w ratios. F-HP polyplexes showed improved cell uptake, lysosomal escape and elevated expression levels of NP in vitro than polyplexes based on HP. Finally, the in-vivo immunization by F-HP/NP polyplexes triggered improved CD8⁺ T cell responses. This study suggests that fluorinated hyperbranched poly (amido amine)s represent one of the effective carriers for DNA vaccine delivery. The animal experiments were approved by the Experimental Animal Ethics Committee of Yangzhou University.

In this study the blending process of Qingyan tablets was simplified and simulated, and near infrared spectroscopy was used to monitor the blending process of several raw materials in different particle size systems to explore the influence of particle size on the blending end-point. Five blended batches with different particle sizes were designed in this experiment and the near infrared spectra of the blended samples were collected. Partial least squares regression (PLSR) models of the contents of Platycodonis Radix, Fructus Chebulae, borax, Hanshuishi and microcrystalline cellulose were developed. A quantitative model was applied to determine the blending end-points of three separate batches of blends with different particle sizes. The moving block of standard deviation (MBSD) was used as a qualitative method for determination of the blending end-point. The results show that the smaller the particle size of the materials, the shorter the time to reach the blending end-points, and with more accurate model predictions. In addition, although the MBSD method is convenient and fast without modeling, the results of blending end-point determination were not as accurate as PLSR method. This approach allowed us to determine the blending end-point of Qingyan tablets.

Heart failure is a serious public health problem with tens of millions of people suffering from its poor prognosis. Epidemiological studies indicate that its morbidity is rising year by year and its mortality has dramatically increased in recent years, with a trend towards younger people. Although great progress has been achieved in the development of medicine in recent years, finding effective agents against heart failure remains an unconquered area in medicine development. A large number of natural medicines and their bioactive compounds, possessing mild and low toxicity as well as multiple target comprehensive effects, have been implicated in a wide range of pharmacological properties in recovering heart function, reducing energy barriers, and improving the life quality of patients. In recent years, they have been widely applied in clinical treatment for heart failure. Hence, summarizing and elucidating the convincing mechanisms for these natural medicines and their bioactive compounds would provide therapeutic targets and benefits for the treatment against heart failure.

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.

Coronavirus disease 2019 (COVID-19), caused by the novel coronavirus (SARS-CoV-2) is treated in accordance with symptoms, which is feasible and effective. However, current therapeutic drugs are ineffective against this virus. The development of targeted therapeutic drugs that are based on key proteins in SARS-CoV-2 replication and pathogenesis will provide a more effective means for clinical treatment. In addition, because SARS-CoV-2 is an RNA virus, which typically mutate readily, new drug development against COVID-19 will be a long-term and arduous task. New approaches to drug discovery for COVID-19 treatment using molecular simulation and machine learning algorithms, and based on the key proteins in the process of SARS-CoV-2 adsorption, entry into the host cell and viral replication are discussed herein, and we briefly introduce related work in our laboratory that can provide strategies to promote the discovery of drugs with different mechanisms of action.

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.

Chloroquine is a quinine derivative which is synthesized by German scholars in 1934. In addition to its anti-malaria, treatment of systemic lupus erythematosus and immunomodulatory effects, chloroquine is also found valuable in broad-spectrum antiviral treatment. Clinical trials have confirmed that chloroquine has a good effect on acquired immunodeficiency syndrome. In 2019, there were many patients infected with novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). Preliminary clinical trials showed that chloroquine had obvious curative effect on patients with SARS-CoV-2. We summarize the effects of chloroquine to different viruses, explain its mechanism, and compare the results of its experiments in vitro and in vivo. The antiviral effect of chloroquine in vivo and in vitro are not consistent, which may be related to the model of animal, dosage and distribution of chloroquine in vivo, and the design of clinical research.

A method was developed for the rapid and systematic identification of alkaloids in Chelidonium majus L. by HPLC-Q-TOF/MS. The separation was performed on an XCharge C18 column (5 μm, 4.6 mm×250 mm) with acetonitrile-0.1% formic acid in water by gradient elution. The flow rate was 0.7 mL·min^-1. The primary mass spectrometer molecular ion and the secondary mass spectrometry fragment ion were determined. The mass spectrometric cleavage of alkaloids was determined and the structures were used to systematically identify the alkaloids in Chelidonium majus L. To verify the accuracy of the qualitative results, three alkaloids were purified and identified by NMR. Twenty-one alkaloids were identified from Chelidonium majus L., including one aporphine-type alkaloid, three protopine-type alkaloids, 11 benzophenanthrine-type alkaloids and six protoberberine-type alkaloids. (S)-N-methylstylopine (8) was first reported in Chelidonium majus L. and dihydrocoptisine (11) and norchelidonine (12) were identified for the first time in Chelidonium majus L. using this technique. The chemical structures of the purified compounds are consistent with the qualitative results of the mass spectrometric analysis. The method is fast and accurate and can provide a basis for the identification and extraction of the chemical constituents of Chelidonium majus L.

The benzaimides chidamide and entinostat are inhibitors of histone deacetylase and have been approved for clinic use. As drug resistance readily occurs in cancer chemotherapy, the characteristics of these drugs were studied in doxorubicin-sensitive and resistant human breast cancer MCF-7 cells. Using a CCK-8 assay for measuring cell proliferation, doxorubicin-resistant cells showed some resistance to chidamide and entinostat, with greater resistance to chidamide. Potentiation of cis-diamine-dichloroplatinum action by entinostat was observed in resistant cells. The accumulation of rhodomine 123, an indirect indicator of ATP-binding cassette B1 (ABCB1)-mediated resistance, was not affected by incubation with chidamide or entinostat, suggesting that neither drug is a substrate for ABCB1. However, ABCB1 expression was significantly increased in resistance cells incubated with a fixed concentration of entinostat. Slowing of the cell cycle at G1 phase and slightly increased cell numbers at G2/M phase was detected by flow cytometry when the cell lines were treated with chidamide or entinostat. Both drugs could induce spherical morphological changes and cleavage of PARP1, an indicater of apoptosis in doxorubicin-sensitive MCF-7 cells, whereas no apoptotic features were observed in resistant cells. These findings show that there is some resistance to chidamide and entinostat in doxorubicin-resistant MCF-7 cells and that this resistance may further oppose apoptosis.

An LC-MS/MS method was developed for the simultaneous determination prednisone acetate, prednisone and active metabolite prednisolone in dog plasma, and applied to a bioavailability and pharmacokinetics study of oral dose of prednisone acetate (2.0 mg·kg^-1) and prednisone (1.8 mg·kg^-1) given to Beagle dogs in a randomized, two-way crossover study. This experiment scheme was approved by the Experimental Animal Ethics Committee of Shanghai Institute of Medicine, Chinese Academy of Sciences. Dexamethason was used as internal standard. After extraction from the plasma by protein precipitation, the analytes and internal standard were separated on an HSS T₃ (50 mm×2.1 mm, 1.8 μm) column using a gradient elution procedure. The mobile phase consisted of methanol and 5 mmol·L^-1 ammonium acetate aqueous solution (0.1% formic acid). Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 401.2→295.2 for prednisone acetate, m/z 359.2→313.2 for prednisone, m/z 361.2→325.1 for prednisolone, m/z 393.2→373.0 for dexamethason. After prednisone acetate was administered, the C_max of prednisone was (25.1 ±3.61) ng·mL^-1 and AUC_0-t was (115 ±27.2) h·ng·mL^-1, while the C_max of prednisolone was (207 ±38.5) ng·mL^-1 and AUC_0-t was (760 ±218) h·ng·mL^-1. After prednisone was administered, the C_max of prednisone was (67.9 ±22.6) ng·mL^-1 and AUC_0-t was (160 ±19.3) h·ng·mL^-1, while the C_max of prednisolone was (582 ±81.4) ng·mL^-1 and AUC_0-t was (1 310 ±140) h·ng·mL^-1. The relative bioavailability of prednisone acetate to prednisone was only 57.1%.