Ligustroflavone mediates the resistance of non-small cell lung cancer to osimertinib by arresting G1/S phase / 药学学报

Natural products are an important source for the development of antitumor lead compounds, but the pharmacological effects and regulatory mechanisms of natural products in osimertinib resistance in non-small cell lung cancer (NSCLC) are not well understood. The natural product ligustroflavone was used as the research object to analyze its efficacy in osimertinib-resistant NSCLC cells by cell proliferation assay and cell cycle detection. The potential targets of ligustroflavone in osimertinib-resistant NSCLC cells were screened by public databases and bioinformatics, molecular docking and microscale thermophoresis were used to identify the interaction between privet and target molecules. Western blot was used to detect the effect of privet on the target molecules and their downstream pathways. Ligustroflavone reduced the proliferation of osimertinib-resistant NSCLC cells, and could arrest the cell cycle. Cyclin-dependent kinase 6 (CDK6) was the potential target of ligustroflavone in osimertinib-resistant NSCLC cells. Ligustroflavone inhibited the activation of CDK6-Rb axis. Together, ligustroflavone could regulate osimertinib resistance in NSCLC cells by binding cell cyclin-related molecules. This study provides a theoretical basis for the targeted drug resistance of NSCLC with natural products, and also provides a new idea for the development of clinical drug combination.

Diagnostic efficacy of indirect haemagglutination assay for detection of Schistosoma japonicum infections among boatmen and fishermen in the Dongting Lake region / 中国血吸虫病防治杂志

OBJECTIVE@#To evaluate the diagnostic efficacy of indirect haemagglutination assay (IHA) for detection of Schistosoma japonicum infections among boatmen and fishermen in Dongting Lake region, so as to provide insights into improving the schistosomiasis surveillance program among boatmen and fishermen.@*METHODS@#The boatmen and fishermen were detected for S. japonicum infections using IHA and Kato-Katz technique or miracidium hatching test nylon gauze simultaneously at schistosomiasis testing sites in the anchor sites for boatmen and fishermen in the Dongting Lake region during the period from 2014 to 2016, and using IHA for serological screening followed by parasitological testing of seropositives during the period from 2017 to 2019. The sensitivity and specificity of IHA were evaluated for detection of S. japonicum infections among boatmen and fishermen, with the 2014-2016 parasitological testing results as a gold standard. In addition, the seroprevalence of S. japonicum infections was compared among boatmen and fishermen with different characteristics and among years.@*RESULTS@#A total of 306 schistosomiasis testing sites were assigned for boatmen and fishermen, and a total of 143 360 person-time boatmen and fishermen were tested for S. japonicum infections in the Dongting Lake region from 2014 to 2019. The sensitivity and specificity of IHA were 69.9%, 97.3% and 96.1% (χ2 = 74.6, P < 0.05), and 70.9%, 74.5% and 71.9% for detection of S. japonicum infections from 2014 to 2016 (χ2 = 29.4, P < 0.05), respectively. The seroprevalence of S. japonicum infections reduced from 30.3% in 2014 to 1.8% in 2019 among boatmen and fishermen, appearing an overall tendency towards a decline (Z = 1 552.4, P < 0.05). In addition, male, individuals at ages of 45 to 60 years, full-time boatmen and fishermen were more likely to be seropositive for S. japonicum infections (all P values < 0.05).@*CONCLUSIONS@#The seroprevalence of S. japonicum infections appeared a tendency towards a decline among boatmen and fishermen in the Dongting Lake region year by year from 2014 to 2019. IHA presented a high efficacy for screening of S. japonicum infections among boatmen and fishermen in the Dongting Lake region.

Recent progress on dissolution and absorption of amorphous solid dispersions / 药学学报

The amorphous solid dispersion is one of the most effective formulation approaches to enhance the oral bioavailability of poorly water-soluble drugs. However, the amorphous drugs tend to crystallize during storage or dissolution due to inadequate formulations, preparation techniques, storage and dissolution conditions, thus negating their advantages. Meanwhile, it is often difficult to establish in vitro-in vivo correlation for amorphous solid dispersions owing to the difference between dissolution media and physiological environments and between the apparent concentration and membrane transport flux, the dynamic process of the in vivo absorption, which put great challenges to the development of amorphous solid dispersion products. This review covers the recent progress on the mechanistic study of the in vitro dissolution and in vivo absorption of amorphous solid dispersions, aiming to provide guidance for the formulation development of poorly soluble drugs.

Immunohistochemical detection of GluA1 subunit of AMPA receptor in the rat nucleus accumbens following cocaine exposure

α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors are differentially regulated in the nucleus accumbens (NAcc) of the brain after cocaine exposure. However, these results are supported only by biochemical and electrophysiological methods, but have not been validated with immunohistochemistry. To overcome the restriction of antigen loss on the postsynaptic target molecules that occurs during perfusion-fixation, we adopted an immersion-fixation method that enabled us to immunohistochemically quantify the expression levels of the AMPA receptor GluA1 subunit in the NAcc. Interestingly, compared to saline exposure, cocaine significantly increased the immunofluorescence intensity of GluA1 in two sub-regions, the core and the shell, of the NAcc on withdrawal day 21 following cocaine exposure, which led to locomotor sensitization. Increases in GluA1 intensity were observed in both the extra-post synaptic density (PSD) and PSD areas in the two sub-regions of the NAcc. These results clearly indicate that AMPA receptor plasticity, as exemplified by GluA1, in the NAcc can be visually detected by immunohistochemistry and confocal imaging. These results expand our understanding of the molecular changes occurring in neuronal synapses by adding a new form of analysis to conventional biochemical and electrophysiological methods.

Immunohistochemical detection of GluA1 subunit of AMPA receptor in the rat nucleus accumbens following cocaine exposure

α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors are differentially regulated in the nucleus accumbens (NAcc) of the brain after cocaine exposure. However, these results are supported only by biochemical and electrophysiological methods, but have not been validated with immunohistochemistry. To overcome the restriction of antigen loss on the postsynaptic target molecules that occurs during perfusion-fixation, we adopted an immersion-fixation method that enabled us to immunohistochemically quantify the expression levels of the AMPA receptor GluA1 subunit in the NAcc. Interestingly, compared to saline exposure, cocaine significantly increased the immunofluorescence intensity of GluA1 in two sub-regions, the core and the shell, of the NAcc on withdrawal day 21 following cocaine exposure, which led to locomotor sensitization. Increases in GluA1 intensity were observed in both the extra-post synaptic density (PSD) and PSD areas in the two sub-regions of the NAcc. These results clearly indicate that AMPA receptor plasticity, as exemplified by GluA1, in the NAcc can be visually detected by immunohistochemistry and confocal imaging. These results expand our understanding of the molecular changes occurring in neuronal synapses by adding a new form of analysis to conventional biochemical and electrophysiological methods.

Pharmaceutical cocrystals: A review of preparations, physicochemical properties and applications

Pharmaceutical cocrystals are multicomponent systems in which at least one component is an active pharmaceutical ingredient and the others are pharmaceutically acceptable ingredients. Cocrystallization of a drug substance with a coformer is a promising and emerging approach to improve the performance of pharmaceuticals, such as solubility, dissolution profile, pharmacokinetics and stability. This review article presents a comprehensive overview of pharmaceutical cocrystals, including preparation methods, physicochemical properties, and applications. Furthermore, some examples of drug cocrystals are highlighted to illustrate the effect of crystal structures on the various aspects of active pharmaceutical ingredients, such as physical stability, chemical stability, mechanical properties, optical properties, bioavailability, sustained release and therapeutic effect. This review will provide guidance for more efficient design and manufacture of pharmaceutical cocrystals with desired physicochemical properties and applications.

Broad-Spectrum Activity of Volatile Organic Compounds from Three Yeast-like Fungi of the Galactomyces Genus Against Diverse Plant Pathogens

The application of antagonistic fungi for plant protection has attracted considerable interest because they may potentially replace the use of chemical pesticides. Antipathogenic activities confirmed in volatile organic compounds (VOCs) from microorganisms have potential to serve as biocontrol agents against pre- and post-harvest diseases. In the present study, we investigated Galactomyces fungi isolated from rotten leaves and the rhizosphere of cherry tomato (Lycopersicon esculentum var. cerasiforme). VOCs produced by Galactomyces fungi negatively affected the growth of phytopathogenic fungi and the survival of nematodes. Mycelial growths of all nine examined phytopathogenic fungi were inhibited on agar plate, although the inhibition was more intense in Athelia rolfsii JYC2163 and Cladosporium cladosporioides JYC2144 and relatively moderate in Fusarium sp. JYC2145. VOCs also efficiently suppressed the spore germination and mycelial growth of A. rolfsii JYC2163 on tomatoes. The soil nematode Caenorhabditis elegans exhibited higher mortality in 24 h in the presence of VOCs. These results suggest the broad-spectrum activity of Galactomyces fungi against various plant pathogens and the potential to use VOCs from Galactomyces as biocontrol agents.

Potentiating CD8

Metabolic regulation has been proven to play a critical role in T cell antitumor immunity. However, cholesterol metabolism as a key component of this regulation remains largely unexplored. Herein, we found that the low-density lipoprotein receptor (LDLR), which has been previously identified as a transporter for cholesterol, plays a pivotal role in regulating CD8

Investigation of oral health knowledge, attitude, behavior of 12-15 years old children in Chongqing / 华西口腔医学杂志

OBJECTIVE@#To investigate the status of oral health knowledge, attitude, behavior of 12-15 years old children and provide a theoretical basis of prevention.@*METHODS@#Multi-stage stratified sampling method was used to extract four middle school students from Chongqing districts and counties (2 in the main urban area and 2 suburbs), and their oral health knowledge, attitudes and behaviors were investigated through questionnaires. All data were entered using Epidata and statistical analysis was performed using SPSS 21.0 software.@*RESULTS@#A total of 3 902 valid questionnaires were collected. The proportion of people who had good brushing habits was 39.7% (1 548), the average oral health knowledge accuracy rate was 58.9%, and the average oral health positive attitude was 88.6%. The number of middle school students who attended the dental experience was 54.5% (2 127), and that of the school who received oral health education was 17.5% (681). There were gender and regional differences in brushing habits.@*CONCLUSIONS@#The knowledge and behavior of oral health among 12-15-year-old middle school students in Chongqing need to be improved. Oral health education for middle school students should be strengthened, especially in rural and suburban areas.

Effect of the local application of stem cells on repairing facial nerve defects: a systematic review / 华西口腔医学杂志

OBJECTIVE@#To systematically evaluate the repairing effect of stem cells on facial nerve defects.@*METHODS@#Articles regarding the regenerating effect of stem cells on facial nerves in animals were collected from the databases of Pubmed, Cochrane Library, Web of Science, Embase, Scopus, and CBM. Two professionals independently completed the article screening, data extraction, and bias risk assessment. RevMan 5.3 and random-effects models were used for the statistical analysis, and the results were presented in the form of mean differences (MD) with a 95%CI. The results of functional evaluation (vibrissae movement, facial paralysis) and histological evaluation (density of myelinated fibers, diameter of fibers, thickness of myelin sheath, G ratio) of facial nerve were Meta-analyzed.@*RESULTS@#A total of 4 614 articles were retrieved from the 6 databases, and 15 of these articles were included in the Meta-analysis. For vibrissae movement and facial paralysis, the stem cell group scored significantly higher than the non-stem cell group (P<0.05). The density of myelinated fibers and thickness of the myelin sheath in the stem cell group were higher than those in the non-stem cell group (P<0.05). The G ratio in the stem cell group was smaller than that in the non-stem cell group (P=0.001). There was no significant difference in fiber diameter (P=0.08).@*CONCLUSIONS@#Stem cells have potential in promoting facial nerve regeneration.

Celastrol inhibits PAK1 kinase and inhibits the proliferation of pancreatic cancer cells / 药学学报

The p21-activated kinase 1 (PAK1) is a member of the P21-activated protein kinase family that plays an important role in the proliferation and on cogenesis of pancreatic cancer. PAK1 is an important target for the treatment of pancreatic cancer. At present, akinase inhibitor targeting PAK1 is still in the preclinical research stage. Therefore, screening for new PAK1 kinase inhibitors is of great significance. In this study the natural compound celastrol was found to have a significant inhibitory effect on PAK1, with an IC50 value of 3.614 μmol·L-1. Molecular docking results showed that celastrol had good binding to PAK1. An MTT assay indicated that celastrol inhibited the proliferation of pancreatic cancer cells BxPC-3 and PANC-1. Mechanistic studies revealed that the inhibition of pancreatic cancer cells by celastrol was reversed by PAK1 siRNA. Celastrol inhibited PAK1 and the subsequent activation of downstream signaling pathways, thereby activating apoptosis signaling pathways and triggering apoptosis in pancreatic cancer cells. These findings suggested that celastrol induced apoptosis in pancreatic cancer cells by suppressing the PAK1 kinase signaling pathway and has potential value for the treatment of pancreatic cancer.

Analysis of a pedigree affected with congenital dysfibrinogenemia due to heterozygous Gln195Arg mutation of fibrinogen γ chain gene / 中华检验医学杂志

Objective:To analyze the phenotype and genotype of a Chinese pedigree with congenital dysfibrinogenemia and investigate the molecular mechanism of the disease.Methods:Pedigree analysis. Peripheral blood samples were collected from 7 members of the pedigree and routine coagulation tests were conducted. The activity of fibrinogen was measured using Clauss method, and fibrinogen antigen was measured by immunoturbidimetry. All the exons and exon-intron boundaries of FGA, FGB and FGG genes were amplified using PCR, which was followed by direct sequencing. Electrophoretic and immunological analysis of fibrinogen, fibrinogen clottability measurement, fibrin polymerization measurement and scanning electron microscopy were used to investigate the pathogenesis of this disease. Results:The proband showed normal activated partial thromboplastin time (APTT) , prolonged prothrombin time(PT), thrombin time (TT),and reptilase time (RT).The antigen level of fibrinogen in the proband (1.6 g/L) decreased slightly, while the activity level of fibrinogen (0.7 g/L) decreased significantly. His father and grandmother showed normal APTT and PT, prolonged TT and RT. The antigen levels of fibrinogen in both of them were normal (2.0 g/L and 2.2 g/L, respectively), while the activity levels of fibrinogen were low (1.0 g/L and 1.1 g/L, respectively). The results of other members from the pedigree were all within the normal range. Genetic analysis revealed a heterozygous A>G mutation at nucleotide 4774 in exon 6 of FGG gene in the proband, which was predicated to be a novel Gln195Arg mutation. The mutation was also found in his father and grandmother.Western blot results showed that no abnormal bands of plasma fibrinogen were found in the proband, his father and grandmother. The fibrinogen clottability in the proband was 49.3%, while that in the heathy control was 98.9%. Both thrombin-induced fibrin polymerization and reptilase-induced fibrin polymerization were significantly impaired in the proband, compared to that in the heathy control. Scanning electron microscopy revealed that compared with the heathy control, the average fiber diameters of the fibrin clot in the proband increased significantly ( P<0.001), while the density of fibers decreased and the arrangement of fibers was sparse. Conclusions:The heterozygous Arg19Gly mutation, which probably damages functions of fibrinogen, should be responsible for the congenital dysfibrinogenemia in this pedigree. This mutation has not been reported.

Ezrin-radixin-moesin proteins are regulated by Akt-GSK3β signaling in the rat nucleus accumbens core

The ezrin-radixin-moesin (ERM) proteins are a family of membrane-associated proteins known to play roles in cell-shape determination as well as in signaling pathways. We have previously shown that amphetamine decreases phosphorylation levels of these proteins in the nucleus accumbens (NAcc), an important neuronal substrate mediating rewarding effects of drugs of abuse. In the present study, we further examined what molecular pathways may be involved in this process. By direct microinjection of LY294002, a PI3 kinase inhibitor, or of S9 peptide, a proposed GSK3β activator, into the NAcc core, we found that phosphorylation levels of ERM as well as of GSK3β in this site are simultaneously decreased. These results indicate that ERM proteins are under the regulation of Akt-GSK3β signaling pathway in the NAcc core. The present findings have a significant implication to a novel signal pathway possibly leading to structural plasticity in relation with drug addiction.

Development of liquiritigenin-phospholipid complex with the enhanced oral bioavailability / 中国天然药物

In the present study, liquiritigenin-phospholipid complex (LPC) was developed and evaluated to increase the oral bioavailability of liquiritigenin. A single-factor test methodology was applied to optimize the formulation and process for preparing LPC. The effects of solvent, drug concentration, reaction time, temperature and drug-to-phospholipid ratio on encapsulation efficiency were investigated. LPCs were characterized by UV-visible spectroscopy, differential scanning calorimetry (DSC), fourier transform infrared spectroscopy (FTIR), and powder X-ray diffractometry (PXRD). The apparent solubility and n-octanol/water partition coefficient were tested. The pharmacokinetic characteristics and bioavailability of the LPC were investigated after oral administration in rats in comparison with liquiritigenin alone. An LPC was successfully prepared. The optimum level of various parameters for liquiritigenin-phospholipid complex was obtained at the drug concentration of 8 mg·mL

Dose selection of chloroquine phosphate for treatment of COVID-19 based on a physiologically based pharmacokinetic model

Chloroquine (CQ) phosphate has been suggested to be clinically effective in the treatment of coronavirus disease 2019 (COVID-19). To develop a physiologically-based pharmacokinetic (PBPK) model for predicting tissue distribution of CQ and apply it to optimize dosage regimens, a PBPK model, with parameterization of drug distribution extrapolated from animal data, was developed to predict human tissue distribution of CQ. The physiological characteristics of time-dependent accumulation was mimicked through an active transport mechanism. Several dosing regimens were proposed based on PBPK simulation combined with known clinical exposure-response relationships. The model was also validated by clinical data from Chinese patients with COVID-19. The novel PBPK model allows in-depth description of the pharmacokinetics of CQ in several key organs (lung, heart, liver, and kidney), and was applied to design dosing strategies in patients with acute COVID-19 (Day 1: 750 mg BID, Days 2-5: 500 mg BID, CQ phosphate), patients with moderate COVID-19 (Day 1: 750 mg and 500 mg, Days 2-3: 500 mg BID, Days 4-5: 250 mg BID, CQ phosphate), and other vulnerable populations (.., renal and hepatic impairment and elderly patients, Days 1-5: 250 mg BID, CQ phosphate). A PBPK model of CQ was successfully developed to optimize dosage regimens for patients with COVID-19.

Approaches used for assessment of the burden of advanced schistosomiasis japonica: a comparative study / 中国血吸虫病防治杂志

Objective To compare the approaches used for the assessment of disability adjust life years (DALYs) for advanced schistosomiasis japonica, so as to provide scientific evidence for accurate assessment of the burden of advanced schistosomiasis japonica. Methods The patients with advanced schistosomiasis japonica receiving treatment and assistance programs in Hunan Province in 2017 were enrolled, and the years lived with disability (YLD) for the patients with advanced schistosomiasis japonica was calculated using the common global burden of disease (GBD) estimation method, the modified GBD method with addition of common syndromes of advanced schistosomiasis japonica, and the quality of life assessment method. Results The YLDs of patients with advanced schistosomiasis japonica, the mean YLDs per capita, and the percentages of YLD were 673.94, 728.77 person-years and 1 761.99 person-years; 0.181, 0.196 person-years and 0.474 person-years; and 10.61, 11.48 person-years per 100 thousand persons and 27.75 person-years per 100 thousand persons with the common GBD method, modified GBD method and the quality of life method, respectively. The YLDs of the patients with advanced schistosomiasis japonica in Hunan Province estimated with the modified GBD method and the quality of life method were 8.14% and 2.61 times higher than that with the common GBD method. Of the major symptoms included in the calculation, the 5 symptoms with the greatest contribution to the burden of advanced schistosomiasis japonica included ascites, moderate anemia, severe anemia, diarrhea and hematochezia. Conclusion The quality of life method may more comprehensively assess the YLDs in patients with advanced schistosomiasis japonica than the common and modified GBD methods.

Approaches used for assessment of the burden of advanced schistosomiasis japonica: a comparative study / 中国血吸虫病防治杂志

Objective To compare the approaches used for the assessment of disability adjust life years (DALYs) for advanced schistosomiasis japonica, so as to provide scientific evidence for accurate assessment of the burden of advanced schistosomiasis japonica. Methods The patients with advanced schistosomiasis japonica receiving treatment and assistance programs in Hunan Province in 2017 were enrolled, and the years lived with disability (YLD) for the patients with advanced schistosomiasis japonica was calculated using the common global burden of disease (GBD) estimation method, the modified GBD method with addition of common syndromes of advanced schistosomiasis japonica, and the quality of life assessment method. Results The YLDs of patients with advanced schistosomiasis japonica, the mean YLDs per capita, and the percentages of YLD were 673.94, 728.77 person-years and 1 761.99 person-years; 0.181, 0.196 person-years and 0.474 person-years; and 10.61, 11.48 person-years per 100 thousand persons and 27.75 person-years per 100 thousand persons with the common GBD method, modified GBD method and the quality of life method, respectively. The YLDs of the patients with advanced schistosomiasis japonica in Hunan Province estimated with the modified GBD method and the quality of life method were 8.14% and 2.61 times higher than that with the common GBD method. Of the major symptoms included in the calculation, the 5 symptoms with the greatest contribution to the burden of advanced schistosomiasis japonica included ascites, moderate anemia, severe anemia, diarrhea and hematochezia. Conclusion The quality of life method may more comprehensively assess the YLDs in patients with advanced schistosomiasis japonica than the common and modified GBD methods.

Scoping review of active surveillance systems for vaccine safety world-wide / 中华预防医学杂志

Objective@#To identify post-marketing active surveillance systems for vaccine safety around the world and understand their features and mechanisms, in order to provide guidance for vaccine administration activities in China.@*Methods@#Following the steps of scoping review, literature about active surveillance system for vaccine safety and published by 30 June 2018 were identified by searching electronic databases, including PubMed, Scopus, and Cochrane Library. Grey literature were also sought by exploring relevant websites. Identified literature were screened according to eligibility criteria, and informative data from included literature were then charted. Framework Synthesis and Thematic Analysis were performed to integrate the charted data.@*Results@#97 pieces of literature were included for review, and 11 active surveillance systems for vaccine safety were identified, mostly located in developed countries. These systems were constructed by 3 types of organizations: administration departments, academic or research institutions, and health care providers. Their data sources included immunization registries, electronic medical records, claims data, case reports of adverse events following immunization electronic questionnaires, and epidemiologic study data. According to their operation procedures, these systems were grouped into 4 modes of active surveillance: Data Linkage, Investigator Network, Automatic Follow-up System, Studies Consortium.@*Conclusion@#Practice of active surveillance for vaccine safety greatly varies across countries, with different conditions and advantages. It is suggested that developing countries should choose suitable mode of active surveillance considering their local situations.

Develop the active surveillance system for vaccine safety in China / 中华预防医学杂志

Post-marketing surveillance of vaccine safety is an important measure to detect adverse events following immunization and therefore reduce the harms to public health. The conventional method for safety surveillance is a passive way through spontaneous reporting, which suffer from under-reporting and incomplete. While active surveillance, a newly proposed surveillance method in developed countries, is capable to make up the deficiencies of passive surveillance. The surveillance system of vaccine safety in China is currently using passive surveillance, and facing many problems and challenges. This arouses a need to promote development of an active surveillance system for vaccine safety in China, learning from the experience world-wide. This commentary aims to throw out suggestions for establishing the active surveillance system, according to the specific situation in China and based on a scoping review of literature.

Advances in coamorphous drug delivery systems

In recent years, the coamorphous drug delivery system has been established as a promising formulation approach for delivering poorly water-soluble drugs. The coamorphous solid is a single-phase system containing an active pharmaceutical ingredient (API) and other low molecular weight molecules that might be pharmacologically relevant APIs or excipients. These formulations exhibit considerable advantages over neat crystalline or amorphous material, including improved physical stability, dissolution profiles, and potentially enhanced therapeutic efficacy. This review provides a comprehensive overview of coamorphous drug delivery systems from the perspectives of preparation, physicochemical characteristics, physical stability, and performance. Furthermore, the challenges and strategies in developing robust coamorphous drug products of high quality and performance are briefly discussed.