The preventive effect and mechanism of Dayuanyin in hypoxic pulmonary hypertension through NF-κB signaling pathway / 药学学报

Dayuanyin (DYY) has been shown to reduce lung inflammation in both coronavirus disease 2019 (COVID-19) and lung injury. This experiment was designed to investigate the efficacy and mechanism of action of DYY against hypoxic pulmonary hypertension (HPH) and to evaluate the effect of DYY on the protection of lung function. Animal welfare and experimental procedures are approved and in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Science. Male C57/BL6J mice were randomly divided into 4 groups: control group, model group, DYY group (800 mg·kg-1), and positive control sildenafil group (100 mg·kg-1). The animals were given control solvents or drugs by gavage three days in advance. On day 4, the animals in the model group, DYY group and sildenafil group were kept in a hypoxic chamber containing 10% ± 0.5% oxygen, and the animals in the control group were kept in a normal environment, and the control solvent or drugs continued to be given continuously for 14 days. The right ventricular systolic pressure, right ventricular hypertrophy index, organ indices and other metrics were measured in the experimental endpoints. Meantime, the expression levels of the inflammatory factors in mice lung tissues were measured. The potential therapeutic targets of DYY on pulmonary hypertension were predicted using network pharmacology, the expression of nuclear factor kappa B (NF-κB) signaling pathway-related proteins were measured by Western blot assay. It was found that DYY significantly reduced the right ventricular systolic pressure, attenuated lung injury and decreased the expression of inflammatory factors in mice. It can also inhibit hypoxia-induced activation of NF-κB signaling pathway. DYY has a protective effect on lung function, as demonstrated by DYY has good efficacy in HPH, and preventive administration can slow down the disease progression, and its mechanism may be related to inhibit the activation of NF-κB and signal transducer and activator of transcription 3 (STAT3) by DYY.

A Network Pharmacology-Based Study on Antidepressant Effect of Salicornia europaea L. Extract with Experimental Support in Chronic Unpredictable Mild Stress Model Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the pharmacodynamic material basis, mechanism of actions and targeted diseases of Salicornia europaea L. (SE) based on the network pharmacology method, and to verify the antidepressant-like effect of the SE extract by pharmacological experiments.@*METHODS@#Retrieval tools including Chinese medicine (CM), PubMed, PharmMapper, MAS 3.0 and Cytoscape were used to search the components of SE, predict its targets and related therapeutic diseases, and construct the "Component-Target-Pathway" network of SE for central nervous system (CNS) diseases. Further, protein-protein interaction (PPI) network, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) function annotation of depression-related targets were analyzed to predict the antidepressant mechanism of SE. Chronic unpredictable mild stress (CUMS) model was used to construct a mouse model with depression-like symptoms. And the animals were randomly divided into 6 groups (n=10) including the normal group (nonstressed mice administered with distilled water), the CUMS group (CUMS mice administered with distilled water), the venlafaxine group (CUMS mice administered with venlafaxine 9.38 mg/kg), SE high-, medium-, and low-dose groups (CUMS mice administered with SE 1.8, 1.35 and 0.9 g/kg, respectively). Then some relevant indicators were determined for experimental verification by the forced swim test (FST), the tail suspension test (TST) and open-field test (OFT). Dopamine (DA) concentration in hippocampus and cerebral cortex, IL-2 and corticosterone (CORT) levels in blood, and nuclear factor E2 related factor 2 (Nrf2), kelch-like epichlorohydrin related protein 1 (Keap1), NAD(P) H dehydrogenase [quinone] 1 (NQO1) and heme oxygenase-1 (HO-1) levels in mice were measured by enzyme linked immunosorbent assay (ELISA) and Western blot respectively to explore the possible mechanisms.@*RESULTS@#The "target-disease" network diagram predicted by network pharmacology, showed that the potential target of SE involves a variety of CNS diseases, among which depression accounts for the majority. The experimental results showed that SE (1.8, 1.35 g/kg) significantly decreased the immobility period, compared with the CUMS group in FST and TST in mice after 3-week treatment, while SE exhibited no significant effect on exploratory behavior in OFT in mice. Compared with CUMS group, the SE group (0.9 g/kg) showed significant differences (P<0.05) in DA levels in the hippocampus and cerebral cortex. In addition, compared with CUMS control group, SE (1.8 g/kg) group showed a significant effect on decreasing the activities of CORT (P<0.05), and serum IL-2 level with no statistical significance. Finally, Western blot results showed that compared with the model group, Nrf2, Keap1, NQO1 and HO-1 protein expressions in SE group (1.8 g/kg) were up-regulated (all P<0.01).@*CONCLUSION@#The SE extract may have an antidepressant effect, which appeared to regulate Nrf2-ARE pathway and increased levels of DA and CORT in the hippocampus and cortex.

Pharyngeal Electrical Stimulation for Post-stroke Dysphagia: A Meta-analysis / 中国康复理论与实践

Objective:To evaluate the effect of pharyngeal electrical stimulation (PES) on post-stroke dysphagia. Methods:Randomized controlled trial (RCT) about pharyngeal electrical stimulation for dysphagia after stroke were searched in Coehrane Library, Embase, EBSCO, PubMed, Web of Science, CBM, VIP, CNKI and Wanfang Data until June, 2020. The literature quality was evaluated, and the data were analyzed with RevMan 5.3. Results:Five RCTs were returned, including 325 patients. PES was more effective in improvement of Dysphagia Severity Rating Scale scores (SMD = -0.27, 95%CI -0.53 to -0.01, P = 0.04) and decannulation rate (RR = 4.69, 95%CI 2.02 to 10.87, P < 0.001); however, there was no significant difference in Functional Oral Intake Scale scores (SMD = 0.24, 95%CI -0.32 to 0.79, P = 0.40), Penetration-Aspiration Scale scores (MD = -0.18, 95%CI -0.74 to 0.39, P = 0.54) and length of stay (SMD = -0.16, 95%CI -0.42 to 0.11, P = 0.25) between PES and control. Conclusion:Pharyngeal electrical stimulation can improve the swallowing function and enhance decannulation rate for post-stroke dysphagia, while it is uncertain for functional oral intake, risk of aspiration and length of stay.

Effects of PRMT5 expression on cell proliferation of ovarian cancer cell HO8910 / 医学研究生学报

Objective Protein arginine methyltransferase 5 (PRMT5),a member of the histone arginine methylation transferase family,is involved in a wide range of biological regulation through ei-ther epigenetic or posttranslational methylation modifications. The pur-pose of the present study was to investigate the effects of PRMT5 on cell proliferation of ovarian cancer cell HO8910. Methods Cell lines HO8910 with PRMT5 overexpression were obtained by transi-ent transfection,which were divided into three groups in the experiment: blank control group (wild-type cell line HO8910),negative control group (HO8910 cells were transfected with pCMV-myc plasmid),and experimental group (HO8910 cells were transfected with pCMV-myc-PRMT5 plasmid). Western blot was used to detect the expression of myc protein,and qRT-PCR was used to detect the ex-pression of PRMT5 mRNA. Cell lines HO8910 with inducible stable knockdown of PRMT5 were established by shRNA interference method,which were divided into four groups: pLKO control group (infected by empty vector lentivirus),pLKO+Dox (100ng/mL) group,shPRMT5 group (infected by PRMT5shRNA lentivirus) and shPRMT5+Dox (100 ng/mL) group. Western blot and qRT-PCR were used to detect the expression of PRMT5 protein and mRNA levels. Dox-induced PRMT5 knockdown was detected by increasing Dox concentration,which includes four groups,Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group,Dox 100ng/mL group,and each group was treated for 48 hours. Western blot and qRT-PCR were used to detect the PRMT5 protein and mRNA expression. Colony formation assay,EdU assay,and CCK-8 assay were used to test cells proliferation. The experiment was conducted in two large groups each with two subgroups: PRMT5 knockdown group (Dox-group,Dox+ group),PRMT5 overexpression group (pCMV-myc group,pCMV-myc-PRMT5 group). Western blot was used to detect the effects of PRMT5 expression on proliferation-related proteins. The experiment was conducted in two large groups,PRMT5 knockdown group with four subgroups : Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group and Dox 100ng/mL group,and PRMT5 overexpression group with two subgroups (pCMV-myc group and pCMV-myc-PRMT5 group). Results Western blot results showed that the expression of myc was detected in the experimental group in which HO8910 cells were transfected with pCMV-myc-PRMT5,and the expression of PRMT5 mRNA was significantly higher in the experimental group than those in the blank control group and the negative control group (P<0.001) . Western blot and qRT-PCR showed that PRMT5 protein (0.32±0.25) and mRNA expression levels in shPRMT5+Dox group were significantly lower than those of shPRMT5 group (0.89±0.18) (P<0.05). Western blot and qRT-PCR confirmed that PRMT5 protein (0.21±0.24) and mRNA expres-sion in Dox 10ng/mL group and Dox 100ng/mL group (0.08±0.15) were significantly downregulated compared to Dox 0ng/mL group (1.11±0.15) (P<0.05). Colony formation experiments,EdU experiments,and CCK-8 experiments confirmed that the proliferative ca-pacity of cells in Dox+group was lower than that of Dox-group in PRMT5 knockdown group(P<0.05); while in PRMT5 overexpression group,the proliferative capacity of pCMV-myc-PRMT5 group was significantly higher than that of the pCMV-myc group (P<0.05). Western blot results showed that the protein expression of Cyclin D1 was significantly lower in Dox 100 ng/mL group (0.17±0.06) than that in Dox 0 ng/mL group (1.18±0.18) (P<0.05) and the expression of P21 was significantly increased in PRMT5 knockdown group (P<0.05). In the PRMT5 overexpression group,the protein expression of Cyclin D1 in pCMV-myc-PRMT5 group (3.48± 0.22) was higher than that in pCMV-myc group (0.88±0.15) (P<0.05),while the protein expression of P21 (0.08±0.17) were significantly lower than that of pCMV-myc group (4.12±0.10) (P<0.05). Conclusion PRMT5 plays an important role in the proliferation of ovarian cancer cells. Down-regulation of PRMT5 can inhibit cell proliferation and up-regulation of PRMT5 can pro-mote cell proliferation.

The mechanism of active ingredients in Salicornia europaea in treatment of diabetes based on pharmacological network / 中成药

AIM To develop a pharmacological network screening method in predicting the potential target,active ingredients and pathway of Salicornia europaea L.for the treatment of diabetes,and to uncover its underlying multi-component,multi-target,multi-pathway mechanism.METHODS Information about fifteen kinds of bioactive chemical constituents of Salicornia europaea L.acquired from a large amount of literature were used to predict the targets according to PharmMapper Server,and such a prediction was also subjected to the screening of the antidiabetes drug targets approved by FDA in the DrugBank database.The relevant information of potential target and pathway was obtained by MAS 3.0 biomolecule function software.Cytoscape software was used to construct the Salicornia europaea L.ingredients-targets-pathways network.RESULTS Fifteen major active ingredients of Salicornia europaea L.affecting in a total of 86 pathways (VEGF signaling pathway,Fc epsilon RI signaling pathway,T cell receptor signaling pathway,etc),including the 30 particular diabetes-related pathways of MAP2K1,MAPK,GSK3B,AKT,etc.,fully demonstrated the multi-component,multi-target,multi-pathway mechanism of Salicornia europaea L.in the treatment of diabetes and its complications,through regulating immune,lipid metabolism,inflammation,apoptosis and other processes.CONCLUSION Given the new understanding in analyzing the scientific connotation of anti-diabetes effect,and the complex system of Salicornia europaea L.,this paper highlights the direction for the next step in the validation experiment of its target and mechanism.

Key problems and methods of cost analysis based on individual cost of patients / 中国卫生政策研究

By summarizing considerations and suggested solutions of domestic and foreign researchers,the gen-eral analysis principles of cost-effectiveness analysis based on patient level data are systematically discussed,as well as the problems that need to be paid attention in the analysis,including the analysis of data set,solution of missing/censored data,consideration of whether discount should be conducted,the use of uncertainty analysis, the prepara-tion of statistical analysis plan,the analysis of intermediate follow-up point data,handling the protocol-driven costs, etc. The paper has provided reference and discussion points for the pharmaceutical economists or readers who may have interest.

LSD1 acetylation mediates TSA-induced apoptosis of ovarian cancer cells / 医学研究生学报

Objective Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide-dependent oxidase, which participates in many biological processes , such as cell proliferation and differentiation and gene activation and repression .The aim of this study was to investigate LSD1 acetylation by histone deacetylase inhib -itor trichostatin A ( TSA) and its effect on TSA-induced apoptosis of ovarian cancer cells . Methods LSD1 shRNA was synthesized and implanted into the pLKO-Tet-On lentiviral vector , which was transfected into HO8910 and SKOV3 ovarian cancer cell lines , and then the transfected cells were screened with 1.5μg/mL puromycin for one week until stable clones were established .The cells were treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), TCP (100μmol/L), or TSA+TCP.And in the experiment of RNA interfering the LSD1 expression, the cells were also treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), Dox (100 ng/mL), or TSA +Dox.The levels of LSD1 acetylation and its substrate histone H3 lysine 4 dimethylation (H3K4me2) were de-tected by immunoprecipitation (IP) and Western blot.The apoptosis of the cells was determined by Annexin Ⅴ/PI staining and flow cytometry, the transcription levels of the Bax and p21 genes detected by real-time quantitative PCR, and the H3K4me2levels in the promoter regions of Bax and p21 measured by chromatin immunoprecipitation ( ChIP ) .Results In comparison with the methanol control, the TSA group showed significantly increased levels of LSD 1 acetylation in the HO8910(1.00±0.29 vs 5.83±0.46, P<0.01) and SKOV3 cells ( 1.00±0.24 vs 5.07±0.35, P<0.01) as well as that of H3K4me2 ( P<0.01);the total apoptosis rates of HO 8910 and SKOV3 cells were remarkably increased in the TSA, TCP, and TSA+TCP groups (P<0.05), even more significantly in the TSA+TCP than in the TSA and TCP groups ( P<0.05) .The mRNA expressions of Bax and p21 in the HO8910 cells were markedly upregulated in the TSA, Dox, and TSA+Dox groups (P<0.05), even more significantly in the latter than in the former two groups (P<0.05).The TSA group exhibited a higher level of H 3K4me2 than the methanol control in the promoters of Bax(2 .92±0.26 vs 0.68±0.19, P<0.01) and p21 (3.07±0.29 vs 0.93±0.17, P<0.01). Conclusion TSA induces the LSD1 acetylation, while suppression of LSD1 expres-sion and activity may enhance the antitumor activity of TSA .

LSD1 acetylation mediates TSA-induced apoptosis of ovarian cancer cells / 医学研究生学报

Objective Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide-dependent oxidase, which participates in many biological processes , such as cell proliferation and differentiation and gene activation and repression .The aim of this study was to investigate LSD1 acetylation by histone deacetylase inhib -itor trichostatin A ( TSA) and its effect on TSA-induced apoptosis of ovarian cancer cells . Methods LSD1 shRNA was synthesized and implanted into the pLKO-Tet-On lentiviral vector , which was transfected into HO8910 and SKOV3 ovarian cancer cell lines , and then the transfected cells were screened with 1.5μg/mL puromycin for one week until stable clones were established .The cells were treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), TCP (100μmol/L), or TSA+TCP.And in the experiment of RNA interfering the LSD1 expression, the cells were also treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), Dox (100 ng/mL), or TSA +Dox.The levels of LSD1 acetylation and its substrate histone H3 lysine 4 dimethylation (H3K4me2) were de-tected by immunoprecipitation (IP) and Western blot.The apoptosis of the cells was determined by Annexin Ⅴ/PI staining and flow cytometry, the transcription levels of the Bax and p21 genes detected by real-time quantitative PCR, and the H3K4me2levels in the promoter regions of Bax and p21 measured by chromatin immunoprecipitation ( ChIP ) .Results In comparison with the methanol control, the TSA group showed significantly increased levels of LSD 1 acetylation in the HO8910(1.00±0.29 vs 5.83±0.46, P<0.01) and SKOV3 cells ( 1.00±0.24 vs 5.07±0.35, P<0.01) as well as that of H3K4me2 ( P<0.01);the total apoptosis rates of HO 8910 and SKOV3 cells were remarkably increased in the TSA, TCP, and TSA+TCP groups (P<0.05), even more significantly in the TSA+TCP than in the TSA and TCP groups ( P<0.05) .The mRNA expressions of Bax and p21 in the HO8910 cells were markedly upregulated in the TSA, Dox, and TSA+Dox groups (P<0.05), even more significantly in the latter than in the former two groups (P<0.05).The TSA group exhibited a higher level of H 3K4me2 than the methanol control in the promoters of Bax(2 .92±0.26 vs 0.68±0.19, P<0.01) and p21 (3.07±0.29 vs 0.93±0.17, P<0.01). Conclusion TSA induces the LSD1 acetylation, while suppression of LSD1 expres-sion and activity may enhance the antitumor activity of TSA .