Inhibitory effect of LZJ541, a novel small molecule inhibitor of STAT3, on the proliferation of hepatocellular carcinoma cells / 药学学报

Signal transducer and activator of transcription 3 (STAT3) is an important regulatory factor of cell proliferation and metastasis, involved in the occurrence and development of a variety of malignant tumors, and it is one of the hot spots in the research of targeted anti-tumor drugs. Our group screened a novel benzobis (imidazole) structure small molecule compound LZJ541 through the screening model of Janus kinase (JAK)/STAT3 pathway inhibitors, which has definite STAT3 inhibitory activity. We examined the effect of LZJ541 on the proliferation of HepG2 and PC-3 cells by MTT assay in vitro, detected the effect of LZJ541 on the expression of STAT3-related proteins in HepG2 cells by Western blot, and measured the effect of LZJ541 on the apoptosis and cell cycle arrest of HepG2 cells via flow cytometry. The results indicated that LZJ541 significantly inhibited the activation of STAT3 signaling pathway and restrained the proliferation of HepG2 cells. Its half maximal inhibitory concentration (IC50) was 13.8 μmol·L-1, which was much lower than that of PC-3 cells (with low STAT3 expression, IC50: 41.99 μmol·L-1), LZJ541 can also inhibit the phosphorylation of STAT3 in HepG2 cells, thereby inducing apoptosis and cycle arrest and then exerting anti-tumor effects. In conclusion, LZJ541 has a certain anti-tumor effect in vitro, which provides an experimental basis for the development of new STAT3-targeted anti-tumor drugs around this kind of compounds.

Validation of compound C19 as a glutaminase C inhibitor / 药学学报

The mitochondrial enzyme glutaminase C (GAC) is highly expressed in a variety of cancer cells, resulting in increased glutamine metabolism and cancer development. Therefore, GAC has become a potential target for anti-tumor drug development. However, current GAC inhibitors shared similar structural characteristics, few new scaffolds were reported. By conducting a prokaryotic Escherichia coli expression system, human GAC protein of high-purity was obtained through lysozyme digestion combined with ultrasound dissociation, and cobalt magnetic beads purification, Moreover, we performed studies to validate interaction between small molecules and GAC protein through thermal shift assay, drug affinity responsive target stability assay, protein crosslinking and GAC enzyme activity detection. Meanwhile, a comprehensive small molecule-protein interaction confirmation and systematic pharmacodynamic study in vitro were carried out on compound C19, which was a reported GAC inhibitor screened from the Enamine database. Results showed that C19 directly bind to GAC protein, disturbed GAC tetramers formation, and inhibited its enzyme catalytic activity. By interfering GAC function, C19 dose-dependently suppressed GAC-mediated glutamine metabolism, reduced glutamate in cancer cells, and thus alleviated A549 and NCI-H1299 non-small cell lung cancer cell growth. Together, C19 was identified as a lead compound, providing a new strategy for the structural design of drugs targeting GAC.

Lipid metabolism and hematological malignancies / 药学学报

With the deepening of research in recent years, tumor metabolic reprogramming has gradually become the focus of research, and targeting tumor cell metabolism has also become a new means of tumor therapy. The metabolic process affects almost all the physiological processes of the organism, and lipid metabolism is an important part of the metabolic process. Studies have shown that changes in lipid uptake, storage and fatty acid synthesis and decomposition have occurred in a variety of tumors. Abnormal lipid metabolism will promote the rapid proliferation of tumors. Abnormal expression of a variety of key metabolic enzymes in the process of lipid metabolism is the key to tumor progression. The purpose of this paper is to explain the metabolic regulation of lipid metabolism and related metabolic enzymes in hematological tumors, and to provide ideas for the treatment of hematological tumors.

Predictive value of preoperative neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio in surgical site infection following radical resection for rectal cancer / 解放军医学杂志

Objective To investigate the predictive value of preoperative neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio in surgical site infection (SSI) following radical resection for rectal cancer. Methods Retrospective analysis of clinical data was performed about 298 patients undergoing radical resection of rectal cancer in the Chinese PLA General Hospital from January 2015 to February 2018. According to whether SSI occurred 30 days after surgery, patients were divided into SSI group (n=20) and control group (n=278). Gender, age, preoperative neoadjuvant chemoradiation, surgical procedure, T stage, and preoperative neutrophil count, lymphocyte count, platelet count, neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), albumin, and hemoglobin level were compared between the two groups. Subgroup analysis was performed in the SSI group according to laparotomy and minimally invasive surgery. Gender, age, preoperative neoadjuvant chemoradiation (CRT), T stage, and preoperative neutrophil count, lymphocyte count, platelet count, NLR, PLR, albumin, and hemoglobin level were compared between two the subgroups. The predictive value of preoperative NLR and preoperative PLR in surgical site infection following radical resection for rectal cancer were analyzed by receiver operating characteristic (ROC) curve. Results The ratio of male to female in the SSI group is higher than control group (19/1 vs. 178/100, P=0.005). There was no significant difference in other demographic and clinical data such as age, non-restorative operation ratio, minimally invasive surgery ratio, neoadjuvant CRT ratio, T stage, preoperative albumin and preoperative hemoglobin between the two groups (P>0.05). There was no significant difference in preoperative neutrophils count [(3.96±1.03)×109/L vs. (3.62±1.28)×109/L, P=0.245], lymphocytes count [(1.47±0.45)×109/L vs. (1.71±0.64)×109/L, P=0.103] and platelets count [(249.10±57.42)×109/L vs. (230.21±68.53)×109/L, P=0.231] between the two groups. Preoperative NLR in the SSI group was significantly higher than that in the control group (2.77±0.52 vs. 2.39±1.23, P=0.010). Preoperative PLR in the SSI group was higher than that in the control group (184.46±69.54 vs. 152.93±73.82, P=0.065), but the difference was not statistically significant. In the SSI group, the age of the laparotomy subgroup was significantly lower than that of the minimally invasive surgery subgroup (49.20±5.54 vs. 61.87±10.24, P=0.018). There was no significant difference in gender, non-restorative operation ratio, neoadjuvant CRT ratio, T stage, preoperative neutrophil count, lymphocyte count, platelet count, NLR, PLR, albumin, and hemoglobin level between two the subgroups (P>0.05). ROC curve analysis showed that AUC of preoperative NLR in prediction of SSI following radical resection for rectal cancer was 0.711(95%CI 0.643-0.779). When the optimal cut off point was 2.13, its sensitivity and specificity was 95.0% and 51.4%, respectively. AUC of preoperative PLR in prediction of SSI following radical resection for rectal cancer was 0.665(95% CI 0.553-0.777). When the optimal cut off point was 150.69, its sensitivity and specificity was 75.0% and 59.7%. Conclusion Preoperative NLR and PLR have predictive value for SSI following radical resection for rectal cancer.

The optimization and application of cell-based screening model for IL-17-mediated signaling pathway / 药学学报

We explore and verify the optimized condition for HEK-Blue IL-17 screening model, and screen the compounds that inhibits IL-17-medited signaling pathway. HEK-Blue IL-17 cells (5×104 cells per well) were seeded into the 96 plates followed by different concentrations of IL-17A or IL-17F alone, or in combination with tested compounds for 16 h. Then, the supernatant medium was incubated with QUANTI-Blue for 1 or 3 h to detect the OD value at λ655nm. The secreted alkaline phosphatase (SEAP) production was an index of IL-17-mediated signaling activation in HEK-Blue IL-17 cells. We found that both IL-17A and IL-17F can significantly activate the IL-17 signaling pathway in HEK-Blue IL-17 cells. The available dosage of IL-17A and IL-17F were 10 and 100 ng·mL-1, respectively. The reaction time of SEAP and QUANTI-Blue was 1 h. In this model, arctigenin and epigallocatechin gallate (EGCG) could inhibit the IL-17A and IL-17F-mediated signaling pathway. This established and optimized screening model of HEK-Blue IL-17 cells was suitable for screening inhibitors of IL-17-mediated signaling pathway.

Effect of Carfilzomib on Proliferation and Apoptosis of Mantle Cell Lymphoma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of Carfilzomib on mantle cell lymphoma (MCL), and to compare with effect of Bortezomib.@*METHODS@#The Jeko-1 cells and primary MCL cells were treated with Carfilzomib for 24, 48 and 72 h, then the inhibitory rate was detected using CCK-8. Lymphocytes derived from healthy volunteer were served as cell controls. Bortezomib and Cyclophosphamide (CTX) were served as medicinal controls. At the same time, the apoptosis of cells treated with different drugs was detected using flow cytometry.@*RESULTS@#The inhibitory effect of Carfilzomib on Jeko-1 cells and primary MCL cells was exhibited with time-dependent and concentration-dependent manners (P＜0.01, r=0.393, r=0.650, rJ=0.473, r=0.417), but the effect on lymphocytes derived from healthy volunteer only showed time-dependence (P＜0.01, r=0.928). Under the same concentration, Carfilzomib exhibited the proliferation Jeko-1 cells stronger than Bortezomib (P＜0.01), but the same inhibition on primary MCL cells was not significantly different from that on lymphocytes derived from healthy volunteer (P＞0.05). Under clinical recommended concentration, Carfilzomib had a stronger inhibitory effect on primary MCL cells than that of Bortezomib (P＜0.01). Cell apoptosis assay showed that under the same concentration the ability of Carfilzomib to induce cell apoptosis was significantly stronger than that of Bortezomib (P＜0.05).@*CONCLUSION@#Carfilzomib can inhibit the growth of MCL cells, its inhibitory rate on the MCL cells is higher than that of Bortezomib.

Adaption and Optimization of Universal Hairpin Transduction in Gene Diagnostics Based on Nucleic Acid Circuits / 分析化学

Isothermal nucleic acid amplifications, as powerful as polymerase chain reaction but functioning at a constant temperature, are considered to be very promising technique in achieving point-of-care gene diagnostics. However, until now, their practical applications are still seriously lagged by the bad reliability resulting from the problems such as false positive amplification and low signal amplitude. In this work, a universal transduction method in which any sequence ( including loop-mediated isothermal amplification products) could be transduced via a hairpin transducer into a catalyst of a well-engineered circuit (catalytic hairpin assembly, CHA) was established. Because CHA circuit could amplify tens to hundreds fold with especially high sequence specificity, it could provide both accuracy and high amplitude for sequence detection. And for a new targeting sequence, the only sequence needed to be changed was the hairpin transducer. Due to the importance of the transducer, we provided and verified a universal designing rule-set to guarantee the transducing efficiency ( signal to background ratio) of the transducer. Transducers designed following this rule set were then proved to be very efficient in detecting pathogen gene targets. As less as near single molecule ( 20 copies ) of pathogen genes could be detected with significant fluorescent and electrochemical signals.

Validation and application a cell-based screening model for IL-6/JAK/STAT3 inhibitor / 药学学报

Interleukin-6 (IL-6)/janus kinase (JAK)/signal transducers and activators of transcription 3 (STAT3) is a pivotal signaling pathway in the regulation of cell proliferation, survival, differentiation and T cell activation. Aberration of this pathway is involved in multiple autoimmunity diseases and cancers, therefore the pathway is considered as a hot target for drug development. In our study, we validated a cell-based model of IL-6/JAK/STAT3 and used it in screening of its inhibitors. HEK-Blue IL-6 cells of Invivogen Inc. were used to stably express IL-6 receptor and STAT3-induced secreted embryonic alkaline phosphatase (SEAP) report gene. After stimulation by IL-6, SEAP was secreted from cells and reacted with QUANTI-Blue. The product can be detected at 655 nm. The inhibitory effect of compounds on STAT3 signaling showed as IC50 was calculated by OD value. The results shown that IL-6 specifically activated the cells, which could be applied to screen the inhibitors for IL-6/JAK/STAT3 signaling pathway. The optimized screening conditions were described as below:50 000 cells/well, 1 ng·mL-1 IL-6 incubation for 20 h and reaction with QUANTI-Blue for 1 h. Based on this condition, we screened 14 natural products based on this cell model and arctigenin, cryptotanshinone and curcumin showed potential inhibitory activities on STAT3 signaling pathway with IC50 of 1.28, 2.96 and 6.61 μmol·L-1. Our study suggests that HEK-Blue IL-6 cells were suitable for screening inhibitors for the IL-6/JAK/STAT3 signaling pathway.

Research progress in the role of heterogeneous nuclear ribonucleoprotein in the central nervous system / 军事医学科学院院刊

Heterogeneous nuclear ribonulcleoprotein(hnRNP)plays a variety of roles in pre-mRNA splicing, nuclear export of mRNA and turnover. It is reported that hnRNP not only takes part in regulating the development of neuron and glial cells, but is closely related to various central nervous system diseases. This paper reviewed the structure, function and the role of hnRNP in the central nervous system in order to provide new insight into the molecule mechanism of nervous system diseases.