Identification of key molecules in miRNA-mRNA regulatory network associated with high-grade serous ovarian cancer recurrence using bioinformatic analysis / 南方医科大学学报

OBJECTIVE@#To investigate the correlation of the potential functional microRNA (miRNA)-mRNA regulatory network with recurrence of high-grade serous ovarian carcinoma (HGSOC) and its biological significance.@*METHODS@#This study was performed based on the data of 354 patients with HGSOC from the Cancer Genome Atlas database. In these patients, HGSOC was divided into different subtypes based on the pathways identified by GO analysis, and the correlations of the subtypes with HGSOC recurrence and differentially expressed miRNAs and mRNAs were assessed. Two relapse-related datasets were identified using the Gene Set Enrichment (GSE) database, from which the differentially expressed miRNAs were identified by intersection with the TCGA data. The target genes of these miRNAs were predicted using miRWalk 2.0 database, and these common differentially expressed miRNAs and mRNAs were used to construct the key miRNA-mRNA network associated with HGSOC recurrence. The expression of miR-506-3p and SNAI2 in two ovarian cancer cell lines was detected using RT-qPCR and Western blotting, and their targeted binding was verified using a double luciferase assay. The effect of miR-506-3p expression modulation on ovarian cancer cell migration was detected using scratch assay and Transwell assay.@*RESULTS@#We screened 303 GO terms of HGSOC-related pathways and identified two HGSOC subtypes (C1 and C2). The subtype C1 was associated with a significantly higher recurrence rate than C2. The differentially expressed genes between C1 and C2 subtypes were mainly enriched in epithelial-mesenchymal transition (EMT). Five miRNAs were identified as potential regulators of EMT, and a total of 41 target genes were found to be involved in the differential expressions of EMT pathway between C1 and C2 subtypes. The key miRNA-mRNA network associated with HGSOC recurrence was constructed based on these 5 miRNAs and 41 mRNAs. MiR-506-3p was confirmed to bind to SNAI2, and up-regulation of miR-506-3p significantly inhibited SNAI2 expression and reduced migration and invasion of SKOV3 and CAOV3 cells (P < 0.05), while miR-506-3p knockdown produced the opposite effects (P < 0.05).@*CONCLUSION@#MiR-506-3p and SNAI2 are the key molecules associated with HGSOC recurrence. MiR-506-3p may affect EMT of ovarian cancer cells by regulating cell migration and invasion via SNAI2, and its expression level has predictive value for HGSOC recurrence.

Study on pharmacoeconomic evaluation of vericiguat in the treatment of heart failure with reduced ejection fraction / 中国药房

OBJECTIVE To evaluate the cost-effectiveness of vericiguat combined with standard treatment in the treatment of heart failure with reduced ejection fraction （HFrEF）. METHODS Based on the results of the VICTORIA trial and related literature， a three-state （including stable state of heart failure， hospitalized state of heart failure and death state） Markov model was constructed. The cycle length was 1 month， the time horizon was 20 years， the discount rate was 5%， and one time China’s per capita gross domestic product （GDP） in 2021 was the willing-to-pay （WTP） threshold. Cost-utility analysis was performed to evaluate the cost-effectiveness of vericiguat combined with standard treatment in the treatment of HFrEF. The output indicators included quality-adjusted life year （QALY） and incremental cost-effectiveness ratio （ICER）. The robustness of the results of the basic analysis was verified by one-way sensitivity analysis and probability sensitivity analysis. RESULTS The ICER of vericiguat combined with the standard treatment plan compared to the standard treatment plan alone was 444 341.95 yuan/QALY， which was more than WTP of this study （80 976 yuan/QALY）. One-way sensitivity analyses showed that the probability of cardiovascular death in both groups was the main influencing parameter for the robustness of the model， but they had little influence on the results of the basic analysis. The probabilistic sensitivity analysis displayed that under the WTP threshold of this study， the possibility of vericiguat combined with the standard treatment plan being more cost-effective was 2.6%. CONCLUSIONS Compared with the standard treatment plan， vericiguat combined with the standard treatment plan is not cost-effective in patients with HFrEF.

Effect of CXCR4 on the Treatment Response and Prognosis of Carfilzomib in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of CXCR4 on the treatment response and prognosis of Carfilzomib (CFZ) in multiple myeloma.@*METHODS@#Dataset GSE69078 based on microarray data from two CFZ-resistant MM cell lines and their corresponding parental cell lines (KMS11-KMS11/CFZ and KMS34-KMS34/CFZ) were downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes (DEGs) were identified, and Protein-protein interaction (PPI) network was established to identify the key genes involved in CFZ resistance acquisition. Finally, the prognostic roles of the CFZ risistance key genes in MM using MMRF-CoMMpass data study was verified.@*RESULTS@#44 up-regulated and 46 down-regulated DEGs were identified. Top 10 hub genes (CCND1, CXCR4, HGF, PECAM1, ID1, HEY1, TCF4, HIST1H4J, HIST1H2BD and HIST1H2BH) were identified via Protein-protein interaction (PPI) network analysis. The CoMMpass data showed that high CXCR4 expression showed correlation to relative higher relapse and progress rates and the overall survival was significant decreased in high CXCR4 patients (P=0.013).@*CONCLUSION@#CXCR4 perhaps plays a crucial role in CFZ acquired resistance, which might help identifying potential CFZ-sensitive patients before treatment and providing a new therapeutic target in CFZ-resistant MM.

Q-markers of Yuquan Capsules based on serum pharmacochemistry of Chinese medicine / 中国中药杂志

This study analyzed the quality markers(Q-markers) of Yuquan Capsules(YQC) based on serum pharmacochemistry of Chinese medicine and detected the components and metabolites of YQC absorbed into the blood by UPLC-Q-TOF-MS and UNIFI systems. As a result, 32 components of YQC were detected, including 17 prototype components and 15 metabolized components. Among them, 12 prototype components(ginsenoside Rh_2, genistein, formononetin, puerarin, daidzein, schizandrin A, schizandrin B, schizandrin C, schizandrol A, schizandrol B, gomisin D, and ononin) and 12 metabolized components(ginsenoside Rg_1, ginsenoside Rg_2, ginsenoside Rg_3, ginsenoside Ro, 3'-methoxypuerarin, daidzin, astragaloside Ⅱ, astragaloside Ⅳ, glycyrrhizic acid, liquiritigenin, isoliquiritin, and verbascoside) showed inhibitory effects and pharmacological activities against diabetes, and these 24 blood-entering components against diabetes were identified as Q-markers of YQC.

Epidemiological and etiological characteristics of Vibrio parahaemolyticus strains causing foodborne disease outbreaks in Guangdong Province from 2017 to 2020 / 中华预防医学杂志

Objective: To study the epidemiological and pathogenic characteristics of Vibrio parahaemolyticus isolated from outbreaks cases in Guangdong Province, 2017-2020. Methods: Epidemiological characteristics of 87 outbreak events caused by Vibrio parahaemolyticus were analyzed. Strains were serotyped, and then analyzed by pulsed-field gel electrophoresis (PFGE). Results: The food-borne disease outbreak caused by Vibrio parahaemolyticus was found in 16 cities. 44.8% (39/87) and 37.9% (33/87) of the outbreaks occurred in hotels, restaurants and school canteens, respectively. Improper food processing and storage (40.2%, 35/87) and cross contamination caused by indiscriminate raw and cooked food (25.3%, 22/87) were the main causes of food-borne disease outbreaks of Vibrio parahaemolyticus. The main serotypes of patient derived strains were O3:K6 (87.5%) and O4:KUT (22.5%). The similarity value between O3:K6 type isolates was 65.5%-100.0%, and the PFGE pattern similarity value of O4:KUT type isolates was 66.5%-100.0%. Conclusion: Outbreaks caused by Vibrio parahaemolyticus are widely distributed in Guangdong province. It is necessary to strengthen the publicity and education on the correct handling of food in hotels, restaurants, schools, and unit canteens. O3:K6 and O4:KUT serotypes are the main serotypes of the outbreak. There is genetic diversity among the epidemic strains.

The predictive value of the distance between the placenta and the internal os of the cervix in second trimester to placenta previa / 中国医师杂志

Objective:To explore the predictive value of the distance between the placenta and the internal os of the cervix (IOD) in second trimester to placenta previa.Methods:476 pregnant women with placenta previa diagnosed by systematic ultrasound in the Affiliated Hospital of North Sichuan Medical College from May 2016 to June 2020 were analyzed retrospectively. The ultrasonic parameters such as IOD, cervical length and placental main attachment position were measured, and the clinical characteristics and pregnancy outcome were recorded. Logistic regression analysis was used to analyze the influencing factors of placenta previa from mild pregnancy to late pregnancy. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of IOD value for placenta previa.Results:197 cases of placenta previa were diagnosed in this study. Multivariate regression analysis showed that the number of previous pregnancies, IOD and history of cesarean section were the related factors of placenta previa from mid pregnancy to late pregnancy ( P<0.05). The risk of placenta previa in pregnant women ≥3 pregnancies was 1.826 times that in pregnant women with less than 3 pregnancies. The risk of placenta previa when the lower edge of placenta covers and crosses the internal orifice of cervix (IOD<0 mm) was 11.494 times that of IOD=0 mm and 22.222 times that of IOD>0 mm<20 mm (low placenta). The risk of placenta previa in pregnant women with a history of cesarean section was 1.908 times that of pregnant women without a history of cesarean section. When the cutoff value of IDO was 20 mm, all pregnant women with placenta previa could be screened out in the group with cesarean section history and the area under the curve (AUC) was 0.840 (95% CI: 0.783-0.896, P<0.05); When the cutoff value of IOD was 13.5 mm, all pregnant women with placenta previa could be screened in the group without cesarean section history, and the AUC was 0.814 (95% CI: 0.759-0.869, P<0.05). Conclusions:The second trimester IOD has a good predictive value for placenta previa.

Physiologically based pharmacokinetic modeling of the inhibitory effect of dapagliflozin on intestinal and renal SGLT / 药学学报

This study establishes and optimizes the physiologically based pharmacokinetics (PBPK) model for dapagliflozin, predicts the drug distribution into relevant tissues, and calculates the inhibitory effect on the sodium-glucose cotransporters (SGLTs) in the intestine and renal proximal tubule. Based on literature data, a PBPK model for oral administration in healthy adults was established and the predicted blood concentration-time curve characteristics, the main pharmacokinetic parameters (PK), and drug excretion in urine were compared with the published data. To verify and optimize the model and verify the accuracy of the tissue distribution and concentration predictions, a pharmacodynamics model (PD) was established. Urine glucose excretion (UGE) was simulated at the corresponding times. The characteristics of the drug-time curve predicted by the model are similar to those of the measured curve, and the ratio of the main PK parameters to the measured values is within a two-fold range; the accuracy of the established PBPK model is good. The maximal inhibition obtained with 10 mg of dapagliflozin on the duodenum and jejunum segment sodium-glucose co-transporter 1 (SGLT1s) was 1.6%-4.7%, and the inhibition rate of the sodium-glucose co-transporter 2 (SGLT2s) in the proximal tubule of the kidney was as high as 99.9%. At a dose of 10 mg, dapagliflozin delayed intestinal glucose absorption while occupying most of the sites (99.9%) of the renal sodium-glucose cotransporter 2 and inhibiting its glucose reabsorption. This physiological-pharmacokinetic model for dapagliflozin in healthy adults can provide meaningful guidance for exploring pharmacological mechanisms and potential toxicity of gliflozin by simulating drug distribution in different tissues.

Exploring the miRNA-mRNA regulatory network in schizophrenia based on GEO database / 四川精神卫生

ObjectiveTo provide a new idea for exploring the molecular genetic approach to the pathogenesis of schizophrenia via construction of microRNA-messenger RNA （miRNA-mRNA） regulatory network in schizophrenia. MethodsThe microarray datasets of GSE54578 miRNA expression profiles in peripheral blood and GSE145554 mRNA expression in the anterior cingulate in postmortem brain of schizophrenic subjects were downloaded from Gene Expression Omnibus （GEO） database since July 2021. The GEO2R was used to identify the differentially expressed miRNAs and mRNAs， screen the miRNA with target differentially expressed mRNA， and predict their potential upstream transcription factors. The overlapping genes from the mRNA targeted by the differentially expressed miRNA and the mRNA differentially expressed in GSE145554 dataset were collected. Then the biological features of hub genes were analyzed via Gene Ontology （GO） analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis， and the protein-protein interaction （PPI） network and miRNA-mRNA regulatory network of hub genes were constructed. ResultsA total of 8 up-regulated differentially expressed miRNAs with targeted mRNA were screened out in GSE54578 datasets regarding schizophrenia， which involved in the regulation of 10 transcription factors， 247 down-regulated differentially expressed mRNAs were screened out in GSE145554 datasets， and 17 overlapping mRNAs were obtained. GO analysis showed that the target mRNAs were mainly involved in astrocyte differentiation and development. KEGG pathway enrichment analysis showed that the target mRNAs were mainly involved in Rap1 and Ras signaling pathways. PPI network analysis showed that the mRNAs （KRAS and CD28） might be key genes in schizophrenia. ConclusionThe integrated bioinformatics analysis based on GEO database can identify potential susceptibility genes in schizophrenia， and it also contributes to the construction of miRNA-mRNA regulatory network in schizophrenia.

VIPR1 promoter methylation promotes transcription factor AP-2α binding to inhibit VIPR1 expression and promote hepatocellular carcinoma cell growth in vitro / 南方医科大学学报

OBJECTIVE@#To explore the transcriptional regulation mechanism and biological function of low expression of vasoactive intestinal peptide receptor 1 (VIPR1) in hepatocellular carcinoma (HCC).@*METHODS@#We constructed plasmids carrying wild-type VIPR1 promoter or two mutant VIPR1 promoter sequences for transfection of the HCC cell lines Hep3B and Huh7, and examined the effect of AP-2α expression on VIPR1 promoter activity using dual-luciferase reporter assay. Pyrosequencing was performed to detect the changes in VIPR1 promoter methylation level in HCC cells treated with a DNA methyltransferase inhibitor (DAC). Chromatin immunoprecipitation was used to evaluate the binding ability of AP-2α to VIPR1 promoter. Western blotting was used to assess the effect of AP-2α knockdown on VIPR1 expression and examine the differential expression of VIPR1 in the two cell lines. The effects of VIPR1 overexpression and knockdown on the proliferation, cell cycle and apoptosis of HCC cells were analyzed using CCK8 assay and flow cytometry. We also observed the growth of HCC xenograft with lentivirus-mediated over-expression of VIPR1 in nude mice.@*RESULTS@#Compared with the wild-type VIPR1 promoter group, co-transfection with the vector carrying two promoter mutations and the AP-2α-over-expressing plasmid obviously restored the luciferase activity in HCC cells (P < 0.05). DAC treatment of the cells significantly decreased the methylation level of VIPR1 promoter and inhibited the binding of AP-2α to VIPR1 promoter (P < 0.01). The HCC cells with AP-2α knockdown showed increased VIPR1 expression, which was lower in Huh7 cells than in Hep3B cells. VIPR1 overexpression in HCC cells caused significant cell cycle arrest in G2/M phase (P < 0.01), promoted cell apoptosis (P < 0.001), and inhibited cell proliferation (P < 0.001), while VIPR1 knockdown produced the opposite effects. In the tumor-bearing nude mice, VIPR1 overexpression in the HCC cells significantly suppressed the increase of tumor volume (P < 0.001) and weight (P < 0.05).@*CONCLUSION@#VIPR1 promoter methylation in HCC promotes the binding of AP-2α and inhibits VIPR1 expression, while VIPR1 overexpression causes cell cycle arrest, promotes cell apoptosis, and inhibits cell proliferation and tumor growth.

Mechanism of Cervi Cornu Pantotrichum in Treatment of Osteoarthritis Based on Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Cervi Cornu Pantotrichumin in the treatment of osteoarthritis by network pharmacology. MethodThe active ingredients and the corresponding targets of Cervi Cornu Pantotrichumin were screened out by a Bioinformatics Analysis Tool of Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）. The targets related to osteoarthritis were obtained through GeneCards and Online Mendelian Inheritance in Man （OMIM）. The targets corresponding to the active ingredients and those related to osteoarthritis were intersected to reveal the common targets， and STRING was adopted to build a protein-protein interaction （PPI） network. DAVID was used for gene ontology （GO） annotation and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment on the anti-osteoarthritis targets of Cervi Cornu Pantotrichumin， and R x64 3.6.3 was employed to produce the advanced bubble charts of GO terms and KEGG pathways. Cytoscape 3.7.2 was used to establish the “Chinese medicinal herb-active ingredient-target-signaling pathway” network. In vitro experiments were performed to detect the viability of RAW 264.7 cells exposed to oxidative stress and the tumor necrosis factor （TNF）-α level in RAW 264.7 cells with inflammation under the treatment by Cervi Cornu Pantotrichumin. ResultA total of 20 active ingredients of Cervi Cornu Pantotrichum were obtained， of which ceramide， 6'-O-β-D-glucosylgentiopicroside， cerebroside， oleuropein， sphingomyelin， and cholesterol ferulate did not meet the screening conditions. Therefore， a total of 14 active ingredients were finally screened out， and 303 and 3 093 targets of active ingredients and osteoarthritis were respectively obtained. The two target sets were taken to intersect， which revealed 92 common targets. GO annotation and KEGG pathway enrichment showed that the targets were mainly involved in redox process， positive regulation of RNA polymerase Ⅱ promoter transcription， inflammatory response， protein synthesis， osteoclast differentiation， TNF signaling pathway， signaling pathways in cancer， mammalian target of rapamycin （mTOR） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and cyclic adenosine monophosphate （cAMP） signaling pathway. The results of in vitro experiments showed that a certain concentration of protein in Cervi Cornu Pantotrichum significantly increased the viability of RAW 264.7 cells exposed to H2O2-induced oxidative damage （P<0.05， P<0.01） and reduced the level of TNF-α in the RAW 264.7 cells experiencing lipopolysaccharide （LPS）-induced inflammation （P<0.05）. ConclusionBased on the network pharmacology method， the mechanism of the multi-component， multi-target and multi-pathway treatment of OA by antler antler was explained， and the anti-inflammatory and antioxidant activities of antler antler were confirmed， which provided theoretical guidance and scientific basis for further research on the treatment of OA by antler antler.

Effect of Fufang Huangbai Fluid Paint on Virulence and Biofilm of Methicillin-resistant Staphylococcus aureus / 中国实验方剂学杂志

ObjectiveTo study the virulence and biofilm inhibition effect of Fufang Huangbai Fluid Paint （FFHBFP） on methicillin-resistant Staphylococcus aureus （MRSA）， and to explore the antibacterial effect of FFHBFP on MRSA， which provides a theoretical basis and reference for clinical medication. MethodFirstly， the microdilution method and time–growth curve were used to determine the minimum inhibitory concentration （MIC） of FFHBFP and vancomycin （VAN） against MRSA and the effect on bacterial growth. The effects of FFHBFP and VAN on the inhibition of MRSA virulence factor lipase and restoration of hydrogen peroxide （H2O2） sensitivity were detected under sub-minimum inhibitory concentration （sub-MIC）. The inhibitory effect of FFHBFP and VAN on MRSA biofilm formation and maturation was detected by the microplate method. The morphological changes of mature biofilms before and after administration were observed under a scanning electron microscope （SEM）. Real-time polymerase chain reaction （Real-time PCR） was utilized to detect the effect of 50.600 g·L-1 concentration of FFHBFP on the expression of MRSA virulence gene crtM and biofilm-forming genes fnbA and icaA. Finally， molecular docking technology was used to predict the mechanism of potential antibacterial active ingredients of FFHBFP in inhibiting the virulence and biofilm of MRSA. ResultThe MIC of VAN was 2 mg·L-1， and VAN below 1 mg·L-1 exerted no effect on MRSA growth. The MIC of FFHBFP was not determined， while the 101.200-202.400 g·L-1 original solution inhibited MRSA growth. Compared with the blank group and the VAN group， sub-MIC （25.300-50.600 g·L-1 original solution） inhibited lipase and recovered MRSA sensitivity to H2O2 （P<0.01）. The results of the microplate method showed that FFHBFP （25.300-202.400 g·L-1 original solution） inhibited biofilm formation and maturation （P<0.05， P<0.01）. The SEM exhibited that FFHBFP made the structure of biofilm loose and the size of the bacteria varied. FFHBFP at 50.600 g·L-1 concentration can inhibit the expression of related virulence genes and biofilm-forming genes （P<0.05， P<0.01）， and molecular docking results also showed that the main antibacterial active ingredients in FFHBFP have good binding ability to the target. ConclusionFFHBFP that cannot directly kill MRSA exerts clinical efficacy by impairing virulence expression， biofilm formation， and other pathogenic properties.

The Effect and Mechanism of Novel Telomerase Inhibitor Nilo 22 on Leukemia Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the cytotoxic effect and its mechanism of the micromolecule compound on the leukemia cells.@*METHODS@#The cytotoxic effects of 28 Nilotinib derivatives on K562, KA, KG, HA and 32D cell lines were detected by MTT assays, and the compound Nilo 22 was screen out. Cell apoptosis and cell cycle on leukemia cells were detected by flow cytometry. The effect of compound screened out on leukemogenesis potential of MLL-AF9 leukemia mice GFP@*RESULTS@#Nilo 22 serves as the most outstanding candidate out of 28 Nilotinib derivatives, which impairs leukemia cell lines, but spares normal hematopoietic cell line. Comparing with Nilotinib, Nilo 22 could induce the apoptosis of GFP@*CONCLUSION@#Nilo 22 shows a significant cytotoxic effect on mice and human leukemia cells, especially for drug resistance cells. Nilo 22 is a promising anti-leukemia agent to solve the common clinical problems of drug resistance and relapse of leukemia.

Establishment of A Mixed Chimeric Mouse Model of Allogeneic Bone Marrow Transplantation and Its Influencing Factors / 中国实验血液学杂志

OBJECTIVE@#To establish a mouse mixed chimerism (MC) model of nonmyeloablative allogeneic bone marrow transplantation(allo-BMT) and explore its affecting factors.@*METHODS@#The MC model was established by nonmyeloablative allo-BMT followed by high-dose post-transplant cyclophosphamide (PTCY). 123 mice in the experiments was retrospectively analyzed, and the factors related with the chimerism were explored with the univariate and multivariate logistic regression analysis. A multivariate linear regression was performed by R project to obtain a mathematical model for predicting the chimeric level with relevant affecting factors.@*RESULTS@#The model presented mixed chimerism on day 14 after transplantation, and was characterized by a donor lymphocyte infusion (DLI) which significantly promoted donor engraftment on day 15, but transfplantation of PBS in control group was failed. Among 123 mice, 47 (38.21%) mice were MC, while 76 (61.79%) mice were non-MC in 123 mice, respectively; univariate analysis showed that the baseline body weight of mice (P=0.001, 17.84±1.19 g vs 18.50±0.94 g), total body irradiation(TBI，P=0.048) and the using of cyclophosphamide (P=0.16) were affected the chimeric state of mice, while the number of infusing cells and the time of detection showed no significant effects. Multivariate regression analysis showed that under certain conditions, the body weight of mice on day 0 was an independent factor affecting chimeric levels (OR=0.493, 95% CI 0.307-0.791, P=0.003). Through R project multiple linear regression, the math model was achieved, which was chimerism=6.09-12×weight(g)+80.03×TBI(Gy)-4.4×cell-counts (× 10@*CONCLUSION@#The experiment presents a method for establishing a mixed chimeric mice model after non-myeloablative bone marrow transplantation and constructs a mathematical model with relevant factors affected chimerism status.

Current research of microinflammation on retinal degenerative diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#Retinal degenerative diseases are kinds of leading cause of blindness characterized by photoreceptor apoptosis and progressive neuronal degeneration. A large body of research has shown the evidence of inflammation reaction in such diseases. Retinal neuroinflammation may be a main factor resulting in apoptosis of photoreceptors and neurodegeneration of retina. In addition, the inflammatory response is not only detected in the retina, but also in aqueous humor, vitreous and even in blood, which forms a persistently very low degree, and chronic inflammatory environment. In this review, we focus on the development of microinflammatory response and its predictor for disease outcomes.

Study on antidepressant effect of total alkaloids of Fibraurea recisa / 中国中药杂志

This study aimed to investigate the antidepressant effects of total alkaloids of Fibraurea recisa in HT22 cells damaged by corticosterone (CORT) in vitro and in a mouse model of chronic unpredictable mild stress (CUMS) as well as the underlying mechanisms.In cellular experiments,the viability of CORT-damaged HT22 cells was detected using cell counting kit-8 (CCK-8),and the cell apoptosis was detected by Hoechst 33258 staining.In animal experiments,C57BL/6N mice were randomly divided into the control group,model group,low (100 mg·kg~(-1)),medium (200 mg·kg~(-1)) and high (400 mg·kg~(-1))-dose of total alkaloids of F.recisa groups,and positive control group.After 21 days of CUMS exposure,their depressive behaviors were observed in behavioral and Morris water maze tests.The serum levels of 5-hydroxytryptamine (5-HT),dopamine (DA),and norepinephrine (NE) were assessed by ELISA.The expression levels of apoptosis-related proteins Bcl-2,Bax and cleaved caspase-3 in HT22 cells and mouse hippocampus were detected by Western blot.The results suggested that total alkaloids of F.recisa alleviated the damage of HT22 cells induced by CORT in a dose-dependent manner.The Hoechst 33258 staining uncovered that total alkaloids of F.recisa better reduced the blue spots and inhibited cell apoptosis.The results of animal experiments showed that total alkaloids of F.recisa significantly improved the depression-like behaviors of mice and increased the serum levels of 5-HT,DA and NE as compared with those in the model group.The Western blot assays revealed a significant up-regulation of Bcl-2 protein expression,but an obvious reduction in Bax and cleaved caspase-3protein expression in the total alkaloids of F.recisa group.In conclusion,total alkaloids of F.recisa inhibited depression possibly by regulating the apoptosis-related protein expression or elevating the monoamine neurotransmitter levels in the brain.

Significance of Targeted Sequencing Assay for Patients with Suspected Myeloid Malignancies / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical significance of the targeted next-generation sequencing assay for patients with suspected myeloid malignancies.@*METHODS@#A total of 39 hematopenia patients with suspected myeloid malignamies in Department of Hematology of The Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University from January 2018 to April 2019 were treated, 20 hot spot genes of myelodysplastic syndrome (MDS) were detected.@*RESULTS@#Regarding the diagnostic type, there were 7 cases of idiopathic cytopenia of undetermined significance (ICUS), 8 cases of clonal cytopenias of undetermined significance (CCUS) and 24 cases of myeloid myeloid malignancies which included 18 cases of MDS, 4 cases of myelodysplastic/myeloproliferative neoplasms (MDS/MPN) and 2 cases of acute myeloid leukemia. Positive mutation was detected in 70.8% (17/24) of myeloid malignancy patients , and 72.7% (16/22) in MDS and MDS/MPN patients. The main mutation types were ASXL1, TET2 and RUNX1. Compared with gene negative group, there were no significant differences in sex, age (＜60 years old or ≥60 years old), proportion of bone marrow blast cells (＜5% or≥5%) and cytogenetics (good, medium and poor) (P＞0.05). Furthermore, all 8 CCUS patients showed positive mutation, and the incidence of double or multiple mutation in CCUS group was significantly lower than that of the MDS and MDS/MPN group (37.5% vs 54.5%) (P=0.002). The mutation types between the two groups were similar, and there was no significant difference in variant allele frequency (P＞0.05).@*CONCLUSION@#Our results suggest that there are high rates of double or multiple mutations in myeloid malignancies, especially in patients with MDS and MDS/MPN. Targeted sequencing assay can improve the diagnosis of myeloid malignancies, and guide clinical treatment.

Survival prognosis and influencing factors of patients with primary liver cancer: a report of 3 106 cases / 中华消化外科杂志

Objective:To investigate the survival prognosis of patients with primary liver cancer and its influencing factors.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 3 106 patients with primary liver cancer who had health insurance for special illness in the Chongqing Malignant Tumor Treatment System from January 2000 to August 2018 were collected. There were 2 559 males and 547 females, aged (60±13)years, with a range from 19 to 95 years. Observation indicators: (1) demographic characteristics; (2) clinical treatment and pathological examination; (3) follow-up and survival; (4) analysis of prognostic factors. Follow-up using telephone interview, outpatient or inpatient reexamination was preformed to detect survival of patients. Follow-up was done once every 3 months within the first year and once a year thereafter up to December 2018. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represent as M (range). Count data were expressed as absolute numbers or percentages. Survival analysis was done after excluding missing data of follow-up. The survival rate was calculated and survival curve was drawn by Kaplan-Meier method. The prognostic factors were analyzed after excluding missing data of follow-up, pathological type, and TNM staging. The log-rank test was used for univariate analysis, and COX proportional hazard model was used for multivariate analysis. Results:(1) Demographic characteristics: of the 3 106 patients with primary liver cancer, the number of males and females (gender), cases with age < 30 years, from 30 to 44 years, from 45 to 59 years, from 60 to 74 years, ≥75 years, cases of Han nationality or other ethnic groups, cases being married or other status (marital status), cases with occupation as enterprise unit staff and (or) workers, public institution personnel and (or) civil servants, freelancers and (or) self-employed entrepreneurs, unemployed, company staff, and other professionals were 2 559, 547, 35, 362, 1 131, 1 163, 415, 3 053, 53, 2 896, 210, 880, 342, 130, 101, 124, and 1 529, respectively. (2) Clinical treatment and pathological examination: of the 3 106 patients with primary liver cancer, cases with hospitalization time < 10 days, from 10 to 19 days, from 20 to 29 days, ≥30 days, cases without surgery or with surgery, cases with hepatocellular carcinoma, cholangiocarcinoma, hybrid type and other pathological types, cases of stage Ⅰ, Ⅱ, Ⅲ, Ⅳ of TNM staging were respectively 771, 1 312, 661, 362, 915, 2 191, 836, 63, 24, 29, 28, 90, 624. There were 2 183 out of 3 106 patients without pathological data and 2 335 without TNM staging data. (3) Follow-up and survival: of the 3 106 patients with primary liver cancer, 2 561 were followed up for 3.0-96.0 months, with a median follow-up time of 27.6 months. The 2 561 patients had survived for 1.0-96.0 months, with a median survival time of 24.7 months. The 1-, 3-, 5-year survival rates were 63.2%, 42.3%, 29.5%, respectively. (4) Analysis of prognostic factors: results of univariate analysis showed that age, marital status, occupation, hospitalization time, surgical treatment, pathological types, and TNM staging were related factors for prognosis of patients ( χ2=31.820, 6.752, 39.100, 120.889, 226.700, 10.452, 48.602, P<0.05). Results of multivariate analysis showed that being married, hospitalization time no less than 30 days, surgical treatment were independent protective factors for prognosis ( hazard ratio=1.463, 0.572, 0.575, 95% confidence interval: 1.044-2.049, 0.413-0.793, 0.438-0.755, P<0.05), stage Ⅲ and Ⅳ of TNM staging were independent risk factors for prognosis of patients ( hazard ratio=3.941, 5.036, 95% confidence interval: 1.687-9.211, 2.237-11.335, P<0.05). Conclusions:Patients with primary liver cancer have poor prognosis. Being married, hospitalization time no less than 30 days, and surgical treatment are independent protective factors for prognosis, stage Ⅲ and Ⅳ of TNM staging are independent risk factors for prognosis.

Construction and in Vitro Study of Glioma-Targeted Cell Membrane Biomimetic Delivery System / 中国药学杂志

OBJECTIVE: To construct a biomimetic delivery system (U251/MSN-DOX), and assess its application of glioma targeted therapy. METHODS: U251 cell membrane was coated on the surface of mesoporous silica nanoparticles(MSN) by co-extrusion to prepare cell membrane biomimetic nanoparticles U251/MSN-DOX. The particle size, potential and morphology were characterized. The physical characteristics, loading content (LC) and encapsulation efficiency (EE) of these nanoparticles were determined. Their toxicity of normal cells was investigated. Their cellular uptake of different formulations in U251 was studied by flow cytometry and fluorescence confocal microscope. Additionally, we assessed the transmembrane transport efficiency of nanoparticles via in vitro BBB. RESULTS: The cell membrane-coated nanoparticles U251/MSN were spherical, and a distinct "core-shell" structure could be observed. The particle size was (135.70±3.85) nm, the LC was (18.57±2.17)%, and the EE was (64.99±2.52)%. The cell experiment showed that U251/MSN had low cytotoxicity and U251/MSN-DOX exhibited stronger cellular uptake ability and BBB transporting efficiency. CONCLUSION: The glioma cell membrane can be coated on the surface of MSN to construct biomimetic nanoparticles U251/MSN. The biomimetic nanoparticles not only are capable of targeting the homologous tumor cells, but also show the enhanced ability to penetrate BBB, which indicate potential applications in the field of tumor targeted drug delivery especially in brain tumor.

Role of pyroptosis in bilirubin-induced microglial injury / 中国当代儿科杂志

OBJECTIVE@#To study whether pyroptosis is involved in the bilirubin-induced injury of primary cultured rat cortical microglial cells.@*METHODS@#Primary cultured rat cortical microglial cells were randomly administered with 30 μmol/L bilirubin (bilirubin group), 30 μmol/L bilirubin following 30 μmol/L VX-765 pretreatment (VX-765+bilirubin group), or an equal volume of dimethyl sulfoxide (control group). Modified MTT assay was used to measure the viability of microglial cells. Western blot was used to measure the expression of the pyroptosis-related proteins Caspase-1 and gasdermin D (GSDMD). Lactate dehydrogenase (LDH)-release assay was used to evaluate the cytotoxicity of microglial cells. EtBr/EthD2 with different molecular weights (394 Da/1 293 Da) was used to measure the size of plasma membrane pores. ELISA was used to measure the level of the inflammatory factor interleukin-1β (IL-1β) in culture supernatant.@*RESULTS@#After bilirubin stimulation, the viability of microglial cells decreased and LDH release increased, both in a time-dependent manner. Compared with the control group, the bilirubin group had a significantly higher positive rate of small-molecule EtBr passing through the cell membrane (P0.05). The expression of activated Caspase-1 significantly increased at 0.5 hour after bilirubin stimulation (P<0.05), and that of activated GSDMD significantly increased at 6 hours after bilirubin stimulation (P<0.05). The release of IL-1β significantly increased at 6 hours after bilirubin stimulation and reached the peak at 24 hours (P<0.001). Compared with the bilirubin group, the VX-765+bilirubin group had a significant increase in cell viability (P<0.05) and significant reductions in the expression of activated GSDMD, the pass rate of EtBr, and the release of LDH and IL-1β (P<0.05).@*CONCLUSIONS@#Pyroptosis is involved in bilirubin-induced injury of primary cultured microglial cells.

Establishment of a New Evaluation Method for Identification of Drug Resistance of Staphylococcus aureus Based on Rapid Identification System of Microorganisms / 中国实验方剂学杂志

Objective::To explore the feasibility of the rapid identification system(MALDI-Biotyper System) of microorganisms for rapid identification of Pseudomonas aeruginosa and clinical isolation of Staphylococcus aureus. Method::Identification quality control and clinical isolation were conducted for drug resistance of S. aureus by microbial rapid identification system and broth dilution method. The scores of microbial rapid identification system were compared with the MIC value of broth dilution method. The drug resistance of P. aeruginosa was simultaneously identified to determine the accuracy and applicability of the rapid identification system of microorganisms. Result::The scores of the microbial rapid identification system showed that the score of sensitive quality control strain S. aureus was higher than 2.000, and the that of resistant strain of methicillin-resistant S. aureus(methicillin-resistant S. aureus, MRSA)was between 1.700 and 2.000.The score of clinically isolated S. aureus was between 1.700 and 2.000, which suggested the drug resistance and was consistent with the MIC value of the broth dilution method. At the same time, the systemic identification value of the P. aeruginosa, which is independent of the quality control sensitive strain, was greater than 2.000, showing sensitivity and it was a sensitive strain itself, which was consistent with the results. Conclusion::The microbial rapid identification system scoring method can be used for the rapid identification of the drug resistance of S. aureus and P. aeruginosa.