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OBJECTIVE@#To determine whether monotropein has an anticancer effect and explore its potential mechanisms against colorectal cancer (CRC) through network pharmacology and molecular docking combined with experimental verification.@*METHODS@#Network pharmacology and molecular docking were used to predict potential targets of monotropein against CRC. Cell counting kit assay, plate monoclonal assay and microscopic observation were used to investigate the antiproliferative effects of monotropein on CRC cells HCT116, HT29 and LoVo. Flow cytometry and scratch assay were used to analyze apoptosis and cell cycle, as well as cell migration, respectively in HCT116, HT29, and LoVo cells. Western blotting was used to detect the expression of proteins related to apoptosis, cell cycle, and cell migration, and the expression of proteins key to the Akt pathway.@*RESULTS@#The Gene Ontology and Reactome enrichment analyses indicated that the anticancer potential of monotropein against CRC might be involved in multiple cancer-related signaling pathways. Among these pathways, RAC-beta serine/threonine-protein kinase (Akt1, Akt2), cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-9 (MMP9), epidermal growth factor receptor (EGFR), cell division control protein 42 homolog (CDC42) were shown as the potential anticancer targets of monotropein against CRC. Molecular docking suggested that monotropein may interact with the 6 targets (Akt1, Akt2, CDK6, MMP9, EGFR, CDC42). Subsequently, cell activity of HCT116, HT29 and LoVo cell lines were significantly suppressed by monotropein (P<0.05). Furthermore, our research revealed that monotropein induced cell apoptosis by inhibiting Bcl-2 and increasing Bax, induced G1-S cycle arrest in colorectal cancer by decreasing the expressions of CyclinD1, CDK4 and CDK6, inhibited cell migration by suppressing the expressions of CDC42 and MMP9 (P<0.05), and might play an anticancer role through Akt signaling pathway.@*CONCLUSION@#Monotropein exerts its antitumor effects primarily by arresting the cell cycle, causing cell apoptosis, and inhibiting cell migration. This indicates a high potential for developing novel medication for treating CRC.
Subject(s)
Humans , Proto-Oncogene Proteins c-akt/metabolism , Cell Proliferation , Matrix Metalloproteinase 9 , Molecular Docking Simulation , Cell Cycle , ErbB Receptors , Apoptosis , Colorectal Neoplasms/pathology , Cell Line, TumorABSTRACT
Objective@#This study aimed to identify internal ribosome entry sites (IRESs) in the open reading frame (ORF) of the Coxsackievirus B3 (CVB3) genome.@*Methods@#The sequences of P1, P2, or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin (ATG) to the end of the P1, P2, or P3 gene were inserted into the pEGFP-N1 vector. After transfection, possible IRES-dependent green fluorescent protein (GFP)-fused proteins were detected by anti-GFP western blotting. The sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors, in which the potential IRESs were determined according to the Fluc/Rluc activity ratio. Expression of Fluc and Rluc mRNA of the bicistronic vector was detected by RT-qPCR.@*Results@#After transfection of full length or truncated sequences of the P1, P2, or P3 plasmids, six GFP-fused protein bands in P1, six bands in P2 and nine bands in P3 were detected through western blotting. Two IRESs in VP2 (1461-1646 nt) and VP1 (2784-2983 nt) of P1; one IRES in 2C (4119-4564 nt) of P2; and two IRESs in 3C (5634-5834 nt) and 3D (6870-7087 nt) of P3 were identified according to Fluc/Rluc activity ratio. The cryptic promoter was also excluded by RT-qPCR.@*Conclusion@#Five IRESs are present in the CVB3 coding region.
Subject(s)
Internal Ribosome Entry Sites/genetics , Open Reading Frames , RNA, Messenger/geneticsABSTRACT
Objective: To explore the correlation between alanine aminotransferase (ALT) trajectories and new-onset metabolic fatty liver disease (MAFLD) to provide a scientific basis for the prevention and treatment of MAFLD. Methods: The study cohort was composed of 3 553 subjects who met the inclusion criteria in the cohort study of the Henan physical examination population. According to the ALT levels of the subjects' physical examination from 2017-2019, three different ALT trajectory groups were determined by R LCTMtools, namely low-stable group, medium-stable group, and high-stable group. The incidence of MAFLD during physical examination in 2020 was followed up, the cumulative incidence rate in each group was calculated by product-limit method, and Cox proportional hazards regression model analyzed the correlation between different ALT trajectories and new-onset MAFLD. Results: The incidence rate of MAFLD parallelly increased with the increase of ALT locus, which was 6.93%, 15.42%, and 19.05%, respectively, and the difference was statistically significant (P<0.001). After adjusting for multiple confounding factors, such as gender, waist circumference, blood pressure, BMI, fasting blood sugar, and blood lipid by Cox proportional hazards regression model, the risks of MAFLD in ALT medium-stable and the high-stable group were still 1.422 times (95%CI:1.115-1.813) and 1.483 times (95%CI:1.040-2.114) of low-stable ALT group (P<0.05). Conclusions: The risk of MAFLD parallelly increases with the increase of ALT level in the normal long-term range. it is necessary to carry out the intervention for MAFLD with long-term average high value to avoid the progress of MAFLD disease to achieve the early prevention on MAFLD.
Subject(s)
Humans , Alanine Transaminase , Body Mass Index , Cohort Studies , Non-alcoholic Fatty Liver Disease/epidemiology , Waist CircumferenceABSTRACT
【Objective】 To investigate the feasibility of using surface electromyography （SEMG） for the detection of abnormal muscle response （AMR） in patients with hemifacial spasm （HFS）. 【Methods】 We retrospectively reviewed the clinical data of HFS patients who underwent microvascular decompression （MVD） in our hospital between June 2019 and December 2020. Patients who received both surface electrode （preoperative） and needle electrode （intraoperative） detection of AMR were included. SEMG recorded from two stimulation-recording sites， namely， zygomatic-mentalis and mandibular marginal-orbicularis oculi， was selected for analyzing the characteristics of AMR. The positive rates of AMR detected by these two kinds of electrodes were comprehensively compared. 【Results】 Totally 77 patients were included in this study. When detected with surface electrodes， the positive rate， latency and amplitude of AMR recorded at zygomatic-mentalis oculi were 90.9% （70/77）， （10.87±1.86） ms and （202.8±47.4） μV， and at mandibular marginal-orbicularis oculi were 92.2% （71/77）， （10.41±1.83） ms and （211.1±54.1） μV， respectively. AMR was detected in 74 patients （96.1%） with surface electrodes. There was no significant difference in positive rate， latency and amplitude of AMR between these two stimulation-recording methods. When detected with needle electrodes， the positive rate of AMR recorded at zygomatic-mentalis oculi was 98.7% （76/77）， which was significantly higher than the rate 89.6% （69/77） recorded at mandibular marginal-orbicularis oculi （P=0.016）. The latency and amplitude of AMR recorded at zygomatic-mentalis were （10.63±1.39） ms and （83.5±27.2） μV， and at mandibular marginal-orbicularis oculi were （10.31±1.18） ms and （58.6±21.4） μV. There was no significant difference in latency between the two stimulation-recording methods， but the amplitude recorded at mandibular marginal-orbicularis oculi was significantly lower （P=0.041）. AMR was detected in 76 patients （98.7%） with needle electrodes. There was no significant difference in the detection rate of AMR between surface electrodes and needle electrodes （P=0.500）， the results were moderately consistent （Kappa=0.490， P<0.001）. 【Conclusion】 The detection efficiency of surface electrodes for AMR is similar to that of needle electrode. With its non-invasive characteristic， the surface electrode can be routinely used for electrophysiological evaluation of HFS.
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Objective:To explore the effects of long non-coding RNA SATB2-AS1 in the proliferation and apoptosis of cervical cancer cells as an endogenous competitive RNA to regulate miR-373-5p/BTG3 axis.Methods:qRT-PCR was used to detect the expression of LncRNA SATB2-AS1, miR-373-5p and BTG3 in paracancerous tissue and cancerous tissue of patients with CC. The interaction between LncRNA SATB2/miR-373-5p/BTG3 was then predicted and verified. The expression of SATB2-AS1 and miR-373-5p in cells was intervened, subsequently the CC cells were divided into different groups. The proliferation activity of cells in each group was detected by MTT, and the apoptosis of cells in each group was detected by flow cytometry.Results:qRT-PCR showed that the expression of SATB2-AS1 and BTG3 in cancer tissue and CC cells was significantly decreased, while the expression of miR-373-5p was significantly increased in cancer tissue and CC cells compared with paracancerous tissue and normal cervical cells.The targeting relationship between SATB2-AS1 and miR-373-5p was confirmed. Compared with NC group, overexpression of SATB2-AS1 inhibited cell proliferation butinduced apoptosis. Overexpression of miR-373-5p promoted cell proliferation, inhibited apoptosis, but this effect was partially saved by SATB2-AS1.Conclusion:Up-regulation of LncRNA SATB2-AS1 expression regulated the miR-373-5p/BTG3 axis and participated in the progression of cervical cancer, subsequently inhibited cancer cell proliferation but induced apoptosis.
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Local focal adhesion kinase (FAK) is a non-receptor intracellular tyrosine kinase that plays an important role in tumor initiation, development, metastasis and invasion, and is considered to be an important target for the development of antineoplastic drugs. It has both kinase-dependent and non-kinase-dependent scaffolding functions. However, traditional small molecular inhibitors can only inhibit its kinase-dependent activity, so it is difficult to target the kinase-independent scaffolding function. Therefore, there is an urgent need for novel strategies to enhance FAK targeting to lay the foundation for determining the druggability and discovery of FAK inhibitors. Proteolysis targeting chimera (PROTAC) is a new drug development strategy that can recruit E3 ligase to specifically ubiquitinylate target proteins for degradation through the proteasome system. The unique mechanism of action of the PROTAC system could be used to target and degrade the FAK protein, thus eliminating the scaffolding function of FAK. In this review, FAK protein, the signaling pathway, and small molecule inhibitors are briefly described, and the latest research progress in targeting the degradation of FAK using PROTAC technology is summarized.
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Objective: To investigate the efficacy and predictors of autologous hematopoietic stem cell transplantation (auto-HSCT) in the treatment of T lymphoblastic lymphoma (T-LBL) . Methods: 41 patients with T-LBL who underwent auto-HSCT from April 2006 to July 2017 in the Department of Hematology, the First Affiliated Hospital of Soochow University and the Department of Lymphoma, Peking University Cancer Hospital were analyzed retrospectively. Results: ①Among 41 patients, there were 30 males and 11 females with median age of 24 (11-53) years old. According to the Ann Arbor staging, 33 (80.5%) patients were in stage Ⅲ/Ⅳ. 12 (29.3%) patients have mediastinal involvement, and 20 (48.8%) patients have bone marrow (BM) involvement. Before transplantation, there were 26 (63.4%) patients who achieved first complete remission (CR(1)) , the other 15 (36.6%) patients were in the non-CR(1) group, and there were 29 (70.7%) patients in the low-intermediate risk group (IPI<3 scores) , the other 12 (34.1%) patients were in the middle-high risk group (IPI≥3 scores) . ②The median follow-up was 29 (3-98) months. The 3-year overall survival (OS) and progression-free survival (PFS) for 41 patients were (64.3±8.2) % and (66.0±7.8) %, respectively. 3-year cumulative recurrence rate (CIR) was (30.7±7.4) %, and 3-year non-recurring mortality (NRM) was (4.8±4.6) %. ③The 3-year OS of the CR(1) group and the non-CR(1) group were (83.4±7.6) % and (38.9±12.9) % (P=0.010) , and the 3-year PFS of two groups were (83.8±7.4) % and (40.0±12.6) % (P=0.006) , respectively. The 3-year CIR of these two groups were (16.2±7.4) % and (53.3±12.9) % (P=0.015) , and the 3-year NRM were 0 and (14.3±13.2) % (P=0.157) , respectively. ④The 3-year OS of the IPI low-intermediate risk group and the high-intermediate risk group were (76.9±8.4) % and (35.7±15.2) % (P=0.014) and the 3-year PFS were (77.4±8.2) % and (40.0±14.6) (P=0.011) , respectively. The 3-year CIR of these two groups were (18.1±7.3) % and (60.0±14.6) % (P=0.006) , and the 3-year NRM were (5.6±5.4) % and 0 (P=0.683) , respectively. The OS and PFS of patients with low-intermediate risk group were significantly higher than the other group. Conclusion: Auto-HSCT could improve the survival of T-LBL. Pre-transplant status and IPI score are important predictors for survival T-LBL patients with auto-HSCT.
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Disease-Free Survival , Hematopoietic Stem Cell Transplantation , Neoplasm Recurrence, Local , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Prognosis , Retrospective Studies , T-Lymphocytes , Transplantation, Autologous , Treatment OutcomeABSTRACT
SUMMARY OBJECTIVE To investigate the clinical efficacy of cefazolin sodium pentahydrate combined with vacuum sealing drainage (VSD) in the treatment of open fracture complicated with soft tissue injury. METHODS Sixty-three patients with open fracture complicated with soft tissue injury were divided into observation (n = 33) and control (n = 30) groups. After surgical reduction, fixation, and repair of the fractures, the control group was treated with VSD for 10 days, and the observation group was treated with cefazolin sodium pentahydrate based on VSD for 10 days. The infection control time was recorded. After treatment, the pain of patients was evaluated. Before and after treatment, the serum levels of C-reactive protein (CRP), interleukin (IL)-6, IL-8, tumor necrosis factor α (TNF-α), cortisol, epinephrine, norepinephrine, and glucose were detected. After 6 months of treatment, the total effective rate of the treatment was evaluated. RESULTS The infection control time and Visual Analogue Scale score after treatment in the observation group were significantly lower than in the control group, respectively (P < 0.05). After the treatment, the serum levels of CRP, IL-6, IL-8, TNF-α, cortisol, epinephrine, norepinephrine, and glucose in each group were significantly lower than before the treatment (P < 0.05), and each index in observation was significantly lower than in the control group (P < 0.05). CONCLUSIONS In the treatment of open fractures complicated with soft tissue injury, cefazolin sodium pentahydrate combined with VSD can effectively reduce inflammation and stress, thus improving the treatment efficacy.
RESUMO OBJETIVO Investigar a eficácia clínica do cefazolin penta-hidrato de sódio combinado com drenagem por vedação a vácuo (VSD) no tratamento da fratura exposta complicada com lesão nos tecidos moles. MÉTODOS Sessenta e três doentes com fratura exposta complicada com lesões nos tecidos moles foram divididos em grupos de observação (n=33) e controle (n=30). Após redução cirúrgica, fixação e reparação da fratura, o grupo de controle foi tratado com VSD durante dez dias e o grupo de observação foi tratado com cefazolina penta-hidrato de sódio com base no VSD durante dez dias. O tempo de controle de infecção foi gravado. Após o tratamento, a dor dos doentes foi avaliada. Antes e após o tratamento, foram detectados os níveis séricos de proteína C-reativa (CRP), interleucina (IL)-6, IL -8, fator de necrose tumoral alfa (TNF-α), cortisol, epinefrina, norepinefrina e glicose. Após seis meses de tratamento, a taxa efetiva total de tratamento foi avaliada. RESULTADOS O tempo de controle da infecção e a pontuação da Escala Visual Analógica após o tratamento no grupo de observação foram significativamente inferiores ao do grupo de controle, respectivamente (P<0,05). Após o tratamento, os níveis séricos de CRP, IL-6, IL-8, TNF-α, cortisol, epinefrina, norepinefrina e glicose em cada grupo foram significativamente menores do que antes do tratamento, respectivamente (P<0,05), e cada índice de observação foi significativamente inferior ao do grupo de controle (P<0,05). CONCLUSÃO No tratamento da fratura exposta complicada com lesões nos tecidos moles, o cefazolin penta-hidrato de sódio combinado com VSD pode efetivamente reduzir a inflamação e o estresse, melhorando assim a eficácia do tratamento.
Subject(s)
Humans , Cefazolin/therapeutic use , Soft Tissue Injuries , Negative-Pressure Wound Therapy , Fractures, Open/therapy , Anti-Bacterial Agents/therapeutic use , Wound Healing , Drainage , Treatment OutcomeABSTRACT
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.
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A simple, sensitive and reliable method was developed for simultaneous determination of ten banned drugs residues including zeranols(ZALs),chloroamphenicol,pentachlorophenol,etc. in swine urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The urine samples were pretreated using lyophilization and QuEChERS procedures, respectively. Acetonitrile and ammonium acetate (5 mmol/L) were chosen as mobile phases. Target compounds were separated well in ZorbaxSB-C18by following the optimized gradient elution program and determined by LC-MS/MS in negative electrospray ionization mode. The linearity of the matrix-matched standard curve of ten analytes in two methods was good in the range of the experimental concentration with correlation coefficients more than 0.99. The recoveries of ten drugs were in the range of 80.7%-107.7% and 73.5%-103.3% at the spiked levels of 5,10 and 20 μg/L by lyophilization and QuEChERS methods,respectively. The coefficients of variation were less than 15%. The limits of detection (LOD) and the limits of quantification (LOQ) from lyophilization and QuEChERS method were 0.1 to 2.0 μg/L and 0.2 to 5.0 μg/L,respectively.
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Objective To investigate the clinical value of multi-slice spiral CT(MSCT) in the diagnosis of mild digestive tract hemorrhage. Methods Thirty-five patients with mild gastrointestinal hemorrhage were examined by enhanced MSCT.CT signs were observed and compared with pathologic examination findings.Results Diseases in these 35 patients included gastric and duodenal ulcer and inflammation(n=4,11.4%),esophagogastric variceal rupture(n=7,20.0%),gastric carcinoma(n=3,8.6%),gastric stromal tumor(n=3,8.6%),gastric polyp(n=1,2.9%),colonic malignancy(n=5,14.2%),small intestinal stromal tumor(n=6,17.1%),rectal cancer(n=2,5.7%),intestinal Crohn disease(n=3,8.6%),and jejunum diverticulum combined with hemorrhage(n=1,2.9%).All of them were diagnosed by endoscopy of the digestive tract and confirmed by operation and pathology.The accuracy rate of CT was 100%.The coincidence rate of CT diagnosis was 97.1%.Conclusion MSCT is a promising technique in the localization,qualitative diagnosis,and clinical treatment of mild gastrointestinal hemorrhage.
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OBJECTIVE To explore the effects and underlying mechanism of Jieyu Anshen granule (JY) in chronic unpredictable mild stress (CUMS)-treated rats after ischemic stroke. METHODS A rat model of post-stroke depression(PSD)was developed by additional CUMS procedures after middle cere-bral artery occlusion(MCAO).Sprague-Dawley rats were given 1 g·kg-1and 3 g·kg-1of JY by gastrogavage for 4 weeks.Escitalopram(10 mg·kg-1)served as a reference drug.Behavioral tests including sucrose preference test, forced swim test and open-field test were performed to evaluate the antidepressant effects. Levels of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) in rat brain were assayed. The anti-inflammatory activity was evaluated by measuring TNF-α and IL-1β in brain. Serum adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were estimated as indices of hypothalamic-pituitary-adrenal (HPA) axis activity. Western blot analysis was used to evaluate hippo-campal expression of the 5-HT1A receptor (5-HT1AR) and brain-derived neurotrophic factor (BDNF). RESULTS PSD rats exhibited decreased sucrose consumption and motor activity, increased immobility time (P<0.01). JY treatment reversed the depressive behaviors in PSD rats (P<0.05, P<0.01). Treat-ment with JY resulted in significantly increased levels of NE, DA and 5-HT in the hippocampus and prefrontal cortex (P<0.05, P<0.01), and increased expression of 5- HT1AR and BDNF in the hippocampus(P<0.01). JY treatment significantly down-regulated the levels of TNF-α and IL-1β in hippocampus andprefrontal cortex (P<0.05). Treatment with JY also resulted in significantly decreased ACTH and CORTin serum which had been increased (P<0.05). CONCLUSION These findings suggest that JY treat-ment could ameliorate PSD, and the effects are likely ascribed to inhibiting HPA axis hyperfunction andinflammatory, up-regulating the levels of neurotransmitters (NE, DA and 5-HT), and the expression ofhippocampal 5-HT1AR and BDNF.
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OBJECTIVE@#To explore the influence of SOX10 on the proliferation and invasion of prostate cancer cells.@*METHODS@#SOX10 protein in prostate cancer cell lines PC3, DU145 and LNcap was detected by Western blotting analysis. The expression of SOX10 in prostate cancer cell lines (PC3 and DU145) were knocked down by small interfering RNAs, and the efficiency of SOX10 by small interfering RNAs was confirmed using Western blotting analysis. CCK-8 assays were conducted to assess the influences of SOX10 on the proliferation of PC3 and DU145 cells, and invasion assays were conducted to assess the influences of SOX10 on the invasion of PC3 and DU145 cells.@*RESULTS@#After SOX10 in prostate cancer cells was knocked down by small interfering RNAs, the proliferation of prostate cancer cells PC3 and DU145 was significantly inhibited. Results of CCK-8 assays showed that the absorbance of PC3 and DU145 in SOX10-silenced groups was decreased compared with those in control groups (PC3: 0 d: 0.166±0.01, 0.162±0.012 vs. 0.155 ±0.01, P>0.05; 1 d: 0.210±0.011, 0.211±0.018 vs. 0.252±0.023, P>0.05; 2 d: 0.293±0.017, 0.280±0.028 vs. 0.433±0.030, P<0.01; 3 d: 0.363±0.071, 0.411±0.038 vs. 0.754±0.045, P<0.01; 4 d: 0.592±0.065, 0.670±0.093 vs. 1.456±0.111, P<0.01. DU145: 0 d: 0.168±0.018, 0.164±0.01 vs. 0.153 ±0.012, P>0.05; 1 d: 0.218±0.007, 0.206±0.024 vs. 0.255±0.02, P>0.05; 2 d: 0.297±0.013, 0.291±0.012 vs. 0.444±0.023, P<0.05; 3 d: 0.378±0.058, 0.419±0.026 vs. 0.762±0.039, P<0.01; 4 d: 0.681±0.094, 0.618±0.050 vs. 1.419±0.170, P<0.01). Meanwhile, knocking down SOX10 significantly suppressed the invasion of prostate cancer cells PC3 and DU145. Results of invasion assays showed that the numbers of invaded cells in SOX10-silenced groups were significantly less than those in control groups (PC3: 142±38, 171±17 vs. 304±55; DU145: 96±22, 134±23 vs. 341±34, P<0.05).@*CONCLUSION@#SOX10 might promote prostate cancer progression by accelerating the ability of the proliferation and invasion of prostate cancer cells, and SOX10 might be a potential therapeutic target for prostate cancer.
Subject(s)
Humans , Male , Cell Line, Tumor , Cell Proliferation , Neoplasm Invasiveness , Prostatic Neoplasms , RNA, Small Interfering , SOXE Transcription Factors/physiologyABSTRACT
OBJECTIVE@#Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3 (CVB3) infection induced autophagy through endoplasmic reticulum (ER) stress.@*METHODS@#In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase (PERK) inhibitor, inositol-requiring protein-1 (IRE1) inhibitor, or activating transcription factor-6 (ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3 (LC3).@*RESULTS@#CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein (GFP)-LC3 punctuation and induced the conversion from LC3-I to phosphatidylethanolamine-conjugated LC3-1 (LC3-II). CVB3 infection still decreased the expression of mammalian target of rapamycin (mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-II to LC3-I in CVB3-infected HeLa cells.@*CONCLUSION@#CVB3 infection induced autophagy through ER stress in HeLa cells, and PERK, IRE1, and ATF6a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.
Subject(s)
Humans , Activating Transcription Factor 6 , Metabolism , Autophagy , Coxsackievirus Infections , Metabolism , Endoplasmic Reticulum Stress , Endoribonucleases , Metabolism , Enterovirus B, Human , HeLa Cells , Protein Serine-Threonine Kinases , Metabolism , Signal Transduction , eIF-2 Kinase , MetabolismABSTRACT
Objective: To evaluate the clinical characteristics and survival outcomes of patients with de novo grade 3 or transformed follicular lymphoma (FL). Methods: Fifty-two patients treated at Peking University Cancer Hospital between January 2009 and September 2017 were assessed, including 28 patients with FL 3A grade, 13 patients with FL 3B grade, 11 patients with transformed FL. Baseline characteristics, survival and prognostic factors were analyzed. Results: ① Twenty-six male and 26 female patients were enrolled, including 28 patients with FL 3A grade, 13 patients with FL 3B grade, 11 patients with transformed FL. ②The 3-year progression-free survival (PFS) and overall survival (OS) for the entire cohort were 56.0% and 80.6%, respectively. Patients with international prognostic index (IPI) score 0-1 demonstrated significantly better 3-year PFS (80.3% vs 20.1%; t=18.902, P<0.001) and OS (95.7% vs 57.0%; t=10.406, P<0.001) than patients with IPI score 2-3. Three-year PFS (94.1% vs 37.2% vs 25.2%; P=0.002) and OS (100.0% vs 76.0% vs 59.8%; P=0.020) were also significantly different among patients with FLIPI 1 score 0-1, 2, ≥3. FLIPI 2 score was also identified as a prognostic factor for 3-year PFS (68.4%, 0, 0; P=0.001) and OS(87.5%, 76.2%, 0; P=0.003). ③Multivariate analysis indicated a significant association of PFS (HR=3.536, P=0.015) and OS (HR=15.713, P=0.015) with IPI. FLIPI 2 was associated with OS (score 0-1, HR=0.078, P=0.007; score 2, HR=0.080, P=0.022). Conclusion: De novo grade 3 or transformed FL might be a group of curable disease with current treatment strategies. IPI is still a prognostic tool in this scenario.
Subject(s)
Female , Humans , Male , Disease-Free Survival , Lymphoma, Follicular , Neoplasm Recurrence, Local , Prognosis , Retrospective Studies , Survival AnalysisABSTRACT
Objective: To evaluate the prognostic value of (18)F-fluorodeoxyglucose positron emission tomography (PET)/computed tomography (CT) in patients with diffuse large B cell lymphoma (DLBCL) undergoing autologous hematopoietic stem cell transplantation (auto-HSCT). Methods: Forty-eight patients with DLBCL treated at Peking University Cancer Hospital between November 2010 and December 2014 were assessed. All patients underwent PET/CT scanning prior to or after auto-HSCT. Correlation analysis was done based upon patients characteristics, PET/CT scan results and survival. Results: ①Among 48 patients, 27 was male, 21 female, median age was 43 (17-59) years old. ② Patients with negative pre-auto-HSCT PET/CT assessment demonstrated significantly better 3-year progression free survival (PFS) (87.1% vs 53.3%, χ(2)=7.02, P=0.019) and overall survival (OS) (90.3% vs 60.0%, χ(2)=6.51,P=0.022) than patients with positive pre-auto-HSCT PET/CT assessment. Three-year PFS (94.1% vs 30.0%, χ(2)=22.75, P=0.001) and OS (97.1% vs 40.0%, χ(2)=21.09, P=0.002) were also significantly different between patients with negative and positive post-auto-HSCT PET/CT assessment. ③ Multivariate analysis indicated a significant association of PFS (HR=13.176, P=0.005) and OS (HR=20.221, P=0.007) with post-auto-HSCT PET/CT assessment. Number of prior treatment regimens was associated with PFS (HR=10.039, P=0.040). ④ Harrell's C index revealed that the value of combined use of number of prior treatment regimens and post-auto-HSCT PET/CT assessment was superior to either one used alone in PFS (Harrell's C values were 0.976, 0.869 and 0.927 in combined use, number of prior treatment regimens and post-auto-HSCT PET/CT assessment, respectively), and the combined use of ECOG performance status and post-auto-HSCT PET/CT assessment significantly increased the Harrell's C index in OS (Harrell's C values were 0.973, 0.711 and 0.919 in combined use, ECOG performance status and post-auto-HSCT PET/CT assessment, respectively). Conclusions: Post-auto-HSCT PET/CT assessment is the main predictor of outcomes in DLBCL patients receiving auto-HSCT. Combined use of post-auto-HSCT PET/CT assessment and number of prior treatment regimens and ECOG performance status is a better prognostic tool in patients with DLBCL undergoing transplantation.
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Disease-Free Survival , Fluorodeoxyglucose F18 , Hematopoietic Stem Cell Transplantation , Lymphoma, Large B-Cell, Diffuse , Positron Emission Tomography Computed Tomography , Prognosis , Retrospective Studies , Transplantation, AutologousABSTRACT
By detecting SRY gene of cell-free fetal DNA ( cffDNA) in maternal peripheral blood, the sex of fetuses was determined, the risks of sex-linked genetic disorders were assessed and the birth rate of sick fetuses was decreased. A method of real-time polymerase chain reaction ( PCR) coupled with invader assay was established to detect SRY gene. This method possessed the advantages such as high sensitivity, high specificity and non-contaminated with closed tube detection. Under the optimized reaction conditions such as 250 nmol/L detection probes, 7. 5 U FEN1 enzyme, 0. 5 U Taq polymerase and 67℃ of annealing temperature in pre-amplification, the simulated samples as low as 4% ( 4 copies/μL ) were detected and two clinical samples with the gestation age of 9 weeks and 10 weeks were successfully detected. The detection results showed that this method could be used to detect SRY gene of cffDNA in maternal peripheral blood, providing an effective technique for clinical non-invasive prenatal diagnosis based on SRY gene.
ABSTRACT
Using four Uncariae Cum Uncis materials including Uncaria sinensis (HGT), U. hirsutea (MGT), Jianhe U. rhynchophylla (JHGT) and U. rhynchophylla(GT) as the research objects, the correlations between medicinal materials' yield and photosynthetic ecophysiology-factors in the plant exuberant growth period were studied. Results showed that the Uncaria plants net photosynthetic rate (Pn) changed by unimodal curve. There was not "midday depression" phenomenon. There was a different relationship among the photosynthetic ecophysiology-factors and between photosynthetic ecophysiology-factors and medicinal materials' yield. Pn,Tl,Gs had a significant correlation with medicinal materials' yield（M）and were the most important factors of growth.
ABSTRACT
By detecting SRY gene of cell-free fetal DNA ( cffDNA) in maternal peripheral blood, the sex of fetuses was determined, the risks of sex-linked genetic disorders were assessed and the birth rate of sick fetuses was decreased. A method of real-time polymerase chain reaction ( PCR) coupled with invader assay was established to detect SRY gene. This method possessed the advantages such as high sensitivity, high specificity and non-contaminated with closed tube detection. Under the optimized reaction conditions such as 250 nmol/L detection probes, 7. 5 U FEN1 enzyme, 0. 5 U Taq polymerase and 67℃ of annealing temperature in pre-amplification, the simulated samples as low as 4% ( 4 copies/μL ) were detected and two clinical samples with the gestation age of 9 weeks and 10 weeks were successfully detected. The detection results showed that this method could be used to detect SRY gene of cffDNA in maternal peripheral blood, providing an effective technique for clinical non-invasive prenatal diagnosis based on SRY gene.
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PURPOSE: To demonstrate the feasibility, safety, and technical strategies of hand-assisted laparoscopic complete mesocolic excision (HAL-CME) and to compare oncological outcomes between HAL-CME and the open approach (O-CME) for right colon cancers. METHODS: Patients who were scheduled to undergo a right hemicolectomy were divided into HAL-CME and O-CME groups. Measured outcomes included demographic variables, perioperative parameters, and follow-up data. Demographic variables included age, sex distribution, body mass index (BMI), American Society of Anesthesiologists (ASA) physical status classification, previous abdominal surgery, tumor localization, and potential comorbidities. Perioperative parameters included incision length, operative time, blood loss, conversion rate, postoperative pain score, postoperative first passage of flatus, duration of hospital stay, total cost, number of lymph nodes retrieved, TNM classification, and postoperative complications. Follow-up data included follow-up time, use of chemotherapy, local recurrence rate, distant metastasis rate, and short-term survival rate. RESULTS: In total, 150 patients (HAL-CME, 78; O-CME, 72) were included. The groups were similar in age, sex distribution, BMI, ASA classification, history of previous abdominal surgeries, tumor localization, and potential comorbidities. Patients in the HAL-CME group had shorter incision lengths, longer operative times, less operative blood loss, lower pain scores, earlier first passage of flatus, shorter hospital stay, higher total costs, similar numbers of lymph nodes retrieved, similar TNM classifications, and a comparable incidence of postoperative complications. The 2 groups were also similar in local recurrence rate, distant metastasis rate, and short-term survival rate. CONCLUSION: The results demonstrate that the HAL-CME procedure is a safe, valid, and feasible surgical method for right hemicolon cancers.