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Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.
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Photothermal therapy (PTT) has attracted significant attention due to minimal side effects and high treatment specificity. However, it often requires very high temperature to achieve complete tumor ablation under a single PTT. Such high temperature brings obvious thermal damage and inflammatory response to the body, affecting the therapeutic effect. In recent years, nitric oxide (NO) has been used to significantly inhibit tumor growth and enhance the sensitivity of tumor cells of temperature and drugs, thus enhancing the therapeutic effect. However, compounds as NO donors often have some disadvantages such as poor biocompatibility and untargeted delivery, etc., therefore, this medical application based on NO therapy is limited. In conclusion, the organic combination of NO donors and photothermal agents (PTAs) is expected to overcome the shortcomings of single therapy and achieve the antitumor effect of "1 + 1 > 2". In view of the rapid development of NO combining with PTT in tumor therapy, this review firstly introduces the antitumor mechanisms of different types of NO donors. Then the treatment strategy based on NO combined with PTT is discussed. Finally, the prospects and challenges of this combination therapy strategy in the clinical treatment of cancer are discussed.
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MicroRNA (miRNA) is a class of endogenous non-coding single-stranded RNA molecules with a length of about 16 ~ 29 nucleotides. Widely found in eukaryotes, they play important regulatory roles in plant cell proliferation and differentiation, organ formation, metabolism, resistance to salt, temperature, drought, heavy metal stress, etc. Plant miRNA mainly affects plant growth and development by degrading target genes or inhibiting the expression of target genes at the translation level. At present, the research on the production and regulation of miRNA is relatively clear, and the specific roles and regulatory networks of miRNA in plant secondary metabolism and response to abiotic stress have also been identified, which lays the foundation for a full understanding of the molecular regulation of miRNA. In order to better understand the expression and regulation characteristics of miRNA and to interpret the regulatory network of miRNA in plant secondary metabolism and abiotic stress response, we review the molecular mechanisms of plant miRNAs in regulating the biosynthesis of various secondary metabolites(flavonoids, terpenoids, alkaloids) and responding to environmental stress (salt, high temperature, low temperature, drought, heavy metal stress),and summarize the regulatory roles of miRNA in secondary metabolism and abiotic stress, which will provide references for understanding the relationship between environmental stress and plant metabolism, for further studying the regulatory mechanism of miRNA in maintaining plant homeostasis, and for cultivating superior crop varieties.
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Wnt signaling pathway can regulate the proliferation and differentiation of neural stem cells, the growth and develop¬ment of neural cells, which is closely related to the therapeutic mechanism of cerebral ischemia-reperfusion injury (CIRI) after ischemic stroke.Researches have showed that some single Chi¬nese medical herb and Chinese medical herb compounds with the effects of tonifying Shen and nourishing Gan, supplementing Qi and nourishing Yin, promoting Xue and Qi, removing blood sta¬sis and resolving phlegm can activate the Wnt signaling pathway in brains and play the role of treating CIRI, and the essence of the treatment is according to the pathogenesis of CIHI which can be reduced by Yin deficiency of Gan and Shen, Qi and Xue de¬ficiency, blood stasis and phlegm toxin.This paper has summa¬rized the research results of Lycium chinensis, Cornus officinalis, turmeric, Salvia miltiorrhiza and other single Chinese medical herbs and Traditional Chinese Medicine(TCM) compounds such as Naoluoxintong, Danlong Xingnao formula, Yiqi Huoxue For¬mula, Bush en Shengsui formula and Huangjing pill in the treat¬ment of CIHI, regulating Wnt signaling pathway in brains.We also have analyzed that the essence of modern medicine of tonify-ing Shen and producing Sui, supplementing Qi and filling Jing in TCM is closely related to promoting cerebral angiogenesis and re¬modeling and stimulating the proliferation and differentiation of neural stem cells in specific areas of the brain after activating Wnt signaling pathway, and promoting blood circulation and re¬moving blood stasis, resolving phlegm and clearing toxin are re-lated to improving brain blood supply, anti-inflammatory, anti- free radical an ti ox id at ion and other functions.
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ObjectiveTo screen the active antitumor components of Gupi Xiaoji decoction by network pharmacology and molecular docking based on the pyroptosis mediated by cysteinyl aspartate-specific protease 1 （Caspase-1） and explore its molecular mechanism in intervening in the pyroptosis of HepG2.2.15 cells through in vitro experiments. MethodThe compounds and targets of Gupi Xiaoji decoction were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） to obtain the corresponding gene symbols. The targets of Caspase-1 were collected from GeneCards，online mendelian inheritance in man（OMIM），PharmGKB，and TTD，and the compound-gene target regulatory network was constructed by Cytoscape. The protein-protein interaction（PPI） network was established and analyzed by STRING. The mechanism of the effective components of Gupi Xiaoji decoction on Caspase-1 was predicted by gene ontology（GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses. The molecular docking was verified with AutoDock Vina. The plasma medicated with Gupi Xiaoji Decoction was prepared and HepG2.2.15 cells were cultured in vitro. HepG2.2.15 cells were divided into a blank plasma group，a VX-765 group，a VX-765+medicated plasma group， and a medicated plasma group. After 48 hours of intervention with 15% medicated plasma， the expression and distribution of gasdermin D-N （GSDMD-N） on the surface of the cell membrane were detected by immunofluorescence staining. The release of lactic dehydrogenase （LDH）， interleukin（IL）-1β，and IL-18 in the cell supernatant was measured by enzyme-linked immunosorbent assay（ELISA） kits. The expression of Caspase-1 and GSDMD-N was measured by Western blot. ResultThe mitogen-activated protein kinase 14 （MAPK14），MAPK1，protein kinase B1 （Akt1）， MAPK8， V-Jun sarcoma virus oncogene homolog （JUN）， and TP53 screened by network pharmacology were the main targets. The compounds 7-hydroxy-5，8-dimethoxy-2-phenyl-chromone，wogonin，rhamnazin，moslosooflavone，isorhamnetin，7-O-methylisomucronulatol，formononetin，calycosin，luteolin，quercetin，kaempferol，β-sitosterol，and baicalein screened by network pharmacology were the main active components of Gupi Xiaoji decoction. Go enrichment analysis showed that multiple biological processes were involved， including responses to oxidative stress and metal ions，ubiquitin-like protein ligase binding，and phosphatase binding. KEGG pathway enrichment analysis showed MAPK pathway，nuclear factor（NF）-κB pathway，p53 pathway， and hypoxia-inducible factor-1（HIF-1） pathway were involved. Molecular docking showed that the targets had good binding with the components. In vitro experiments displayed that compared with the blank plasma group，the VX-765 group showed weakened GSDMD-N fluorescence signal，reduced release of LDH，IL-1β，and IL-18，and declining expression of Caspase-1 and GSDMD-N（P<0.01）， and the medicated plasma group showed increased GSDMD-N fluorescence signal， increased release of LDH，IL-1β，and IL-18，and up-regulated expression of Caspase-1 and GSDMD-N（P<0.01）. ConclusionGupi Xiaoji Decoction can induce the pyroptosis of HepG2.2.15 cells by regulating Caspase-1 through multiple targets and multiple pathways.
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OBJECTIVE@#To investigate the efficacy, safety and the risk factors affecting prognosis of high-risk acute myeloid leukemia (AML) patients treated by cladribine-based intensified conditioning regimen.@*METHODS@#The clinical data of 28 patients with high-risk AML treated by cladribine in combination with busulfan plus cyclophosphamide (BuCy) intensified conditioning regimen before allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Zhujiang Hospital, Southern Medical University from October 2016 to June 2020 were analyzed retrospectively. The overall survival (OS) rate, cumulative progression-free survival (PFS) rate, relapse rate, non-relapse mortality (NRM), regimen related toxicity (RRT) and risk factors affecting prognosis of the patients were analyzed.@*RESULTS@#The 1-year OS and PFS of the patients after implantation was (78.8±8.6)% and (79.8±8.1)%, while the 1-year cumulative relapse rate and NRM of the patients was 9.3% and 22.0%, respectively. The 1-year expected OS of MRD- high-risk patients before HSCT was 100%. The 1-year expected OS and PFS of the patients in pre-transplant relapse group was (46.9±18.7)% and (50.0±17.7)%, respectively. The incidence of I/II grade RRT was 39.3%. NO III/IV grade RRT were found in 28 patients. Multivariate analysis showed that pre-transplant relapse was the independent risk factor affecting OS and PFS of the patients.@*CONCLUSION@#The intensified conditioning regimen of cladribine in combination with BuCy can reduce the relapse rate of high-risk AML transplantation, and its RRT is mild, exhibiting good safety. MRD- high-risk patients before HSCT can achieve better transplant benefits, but the prognosis of patients with relapse before transplantation is not significantly improved. Therefore, for non-relapsed high-risk AML patients, this intensified conditioning regimen deserves to be considered.
Subject(s)
Humans , Busulfan , Cladribine , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Leukemia, Myeloid, Acute/therapy , Retrospective Studies , Transplantation ConditioningABSTRACT
Objective:DNA barcodes suitable for Lauraceae plants were screened，and 22 Lauraceae plants were identified and classified. Method:The DNA of 22 species of Lauraceae was extracted and amplified by polymerase chain reaction（PCR） with different DNA barcoding primers，followed by electrophoresis and sequencing. Codon Code Aligner was used to proofread，splice， and remove the forward and reverse primer sequences. The sequence was imported into DNAMAN for multiple sequence alignment，and the base mutation sites were analyzed. The Kimura 2-Parameter（K2P） distance of different plants was calculated by MEGA，and the variation degree was analyzed according to the genetic distance. The phylogenetic tree was constructed based on the adjacency method. Result:Three pairs of DNA barcoding primers were used to amplify and sequence the DNA of 22 species of Lauraceae，and 20 species were identified by comparing the specific base sites of gene sequences<bold>.</bold> Conclusion:Four <italic>Litsea</italic> plants could be identified by <italic>matK</italic>， three <italic>Phoebe</italic> plants except for<italic> Cinnamomum validinerve </italic>by<italic> rbcL</italic>， and 20 Lauraceae plants by the combination of<italic> matK</italic>， <italic>rbcL</italic>， and <italic>rpoB</italic>，which provided a theoretical basis for the identification and development of Lauraceae plants. Among them，<italic>matK</italic> was predominant in the identification of Lauraceae plants，and the results also showed that the combination of sequences for plant identification could achieve a better result in DNA barcoding. This study investigated the genetic relationship between Lauraceae plants at the molecular level，aiming at providing a basis for the investigation，cultivation，development，protection， and utilization of medicinal plant resources.
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Exosomes are involved in invasion, migration and angiogenesis of tumor cells, and invasion is the main cause of death in glioma patients. Studies have shown that the exosomes secreted by tumor cells can carry miRNA into the receptor cells and regulate the biological functions of the receptor cells, such as proliferation, migration and invasion. miR-574-5p plays a key role in the occurrence and development of a variety of tumors. However, whether the exosomes derived from glioma cells express miR-574-5p and its role in the growth, invasion and migration of glioma cells have not been reported. This study investigated the mechanism of the exosomal miR-574-5p secreted from glioma cells in the process of cell proliferation, migration and invasion. The exosomes were characterized by electron microscopy, nanoparticle size tracking and Western blot. The results displayed that the extracted exosomes were round particles with a diameter of 30 ~ 100 nm. The internalization of exosomes was detected by immunofluorescence assay. The results showed that exosomes were internalized into LN229 cells; Bioinformatics and online data were used to screen the differentially secreted miRNA between LN229 and H4 glioma cells. The results showed that the differentially secreted miRNA was miR-574-5p, and large tumor suppressor 2 (LATS2) was predicted to be the target gene of miR-574-5p; Duel luciferase reporter assay confirmed that miR-574-5p was complementary to the 3'UTR region of LATS2; The transfection assay, qRT-PCR and Western blot was conducted to measure the relationship between miR-574-5p and LATS2. The results demonstrated that there was no significant difference in LATS2 mRNA levels between the control group and the group with miR-574-5P overexpression (P > 0.05), suggesting the regulatory effect of miR-574-5P on LATS2 was achieved by inhibiting its translation (P < 0.05). CCK-8, Transwell migration and invasion assays were conducted to explore the effect of miR-574-5p on proliferation, migration and invasion of LN229 cells. The results showed that overexpression of miR-574-5p could significantly promote the ability of proliferation, migration and invasion of LN229 cells (P < 0.05). In addition, Western blot was performed to measure the expression of kinase proteins involved in the LATS2/YAP signaling pathway, and the influence of the exosomes on this signaling pathway. The results revealed that the exosomes down-regulated the protein expression level of LATS2 and reduced p-YAP phosphorylation. In conclusion, the exosomal miR-574-5p can promote the proliferation, migration and invasion of glioma cells by down-regulating LATS2 and activating LATS2/YAP signaling pathway, which may provide a potential biomarker for the diagnosis and target for the treatment of glioma.
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OBJECTIVE@#To investigate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of acute monocytic leukemia (AML-M5) and the related factors that affecting the prognosis of the patients.@*METHODS@#The clinical data of 71 patients with AML-M5 treated with allo-HSCT in Zhujiang Hospital Affiliated to Southern Medical University from April 2009 to October 2019 were collected and retrospectively analyzed. The incidence of graft-versus-host disease (GVHD), cumulative overall survival (OS) rate, cumulative progression-free survival (PFS) rate, transplantation-related mortality (TRM), relapse rate and the risk factors affecting prognosis in the patients were analyzed.@*RESULTS@#66 patients obtained hematopoietic reconstruction after transplantation, the median time of granulocyte implantation was 12 (9-26) d, and the median time of megakaryocytic implantation was 13 (8-72) d. The incidence of acute GVHD and chronic GVHD was 33.8% (24/71) and 36.6% (26/71), respectively. The median follow-up time was 13.81 (0.16 to 112.54) months; the median OS and PFS was 31.27 and 26.07 months, respectively. The cumulative OS of the patients in 1 and 3 years after transplantation was 64.9% and 48.6%, respectively, and the cumulative PFS of the patients in 1 and 3 years was 55.0% and 39.5%, respectively. The cumulative relapse rate of the patients in 1 and 3 years was 24% and 40%, respectively. Multivariate analysis showed that pre-transplantation relapse was the independent risk factor affecting OS (HR=2.32, 95%CI:1.17-4.62, P=0.02) and PFS (HR=3.08, 95%CI:1.61-5.90, P=0.001) of the patients; invasive fungal disease after transplantation was the independent risk factor affecting OS (HR=2.71, 95% CI:1.32-5.56, P=0.007) and PFS (HR=2.87, 95%CI=1.40-5.86, P=0.004) of the patients; FLT3 mutation was the independent risk factor affecting PFS (HR=2.13, 95%CI=1.07-4.24, P=0.03) of the patients.@*CONCLUSION@#AML-M5 is the intermediate or high-risk leukemia, and allo-HSCT can improve the survival prognosis of the patients. Pre-transplantation relapse and invasive fungal disease after transplantation are the important factors affecting the efficacy of allo-HSCT in patients with AML-M5.
Subject(s)
Child , Humans , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Leukemia, Monocytic, Acute , Leukemia, Myeloid, Acute , Patients , Prognosis , Retrospective StudiesABSTRACT
<p><b>AIM</b>To study bioequivalences of bambuterol and its metabolites terbutaline in 20 healthy male volunteers.</p><p><b>METHODS</b>A single oral dose of domestic bambuterol capsule or imported bambuterol tablet was given according to a randomized 2-way cross-over design. The plasma bambuterol and terbutaline concentrations were determined by high performance capillary zone electrophoresis (HPCZE).</p><p><b>RESULTS</b>The pharmacokinetic parameters of the capsule and tablet of bambuterol: AUC0-1 were (72 +/- 18) and (72 +/- 13) microgram.h.L-1, Cmax were (8.1 +/- 1.8) and (9.2 +/- 2.3) microgram.L-1, Tmax were (3.6 +/- 1.3) and (3.7 +/- 1.0) h, respectively; terbutaline: AUC0-t were (129 +/- 32) and (130 +/- 34) microgram.h.L-1, Cmax were (7.8 +/- 2.2) and (8.5 +/- 2.9) microgram.L-1, Tmax were (5.4 +/- 0.8) and (5.6 +/- 1.1) h, respectively. The bioavaiability of the capsule was (100 +/- 16)% (bambuterol) and (101 +/- 13)% (terbutaline).</p><p><b>CONCLUSION</b>The results demonstrated that the two preparations of bambuterol and terbutaline were bioequivalent by analysis of variance, with two-one sided test at 90% confidential level.</p>