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1.
Biomed Pharmacother ; 121: 109310, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31710895

RESUMO

Currently, there is no effective method to prevent renal interstitial fibrosis after acute kidney injury (AKI). In this study, we established and screened a new renal interstitial fibrosis rat model after cisplatin-induced AKI. Our results indicated that rats injected with 4 mg/kg cisplatin once a week for two weeks after firstly administrated with 6.5 mg/kg loading dose of cisplatin could set up a more accurate model reflecting AKI progression to renal interstitial fibrosis. Then, we investigated the effects and possible mechanisms of human umbilical cord blood mononuclear cells (hUCBMNCs) on renal tubular interstitial fibrosis after cisplatin-induced AKI. In rats injected with hUCBMNCs for four times, level of matrix metalloproteinase 7(MMP-7)in serum and urine, urinary albumin/creatinine ratio, tubular pathological scores, the relative collagen area of the tubulointerstitial region, endoplasmic reticulum dilation and the mitochondrial ultrastructural damage were significantly improved. The level of reactive oxygen species, α-smooth muscle actin (α-SMA), [NOD]-like pyrin domain containing protein 3 and cleaved-Caspase 3 in renal tissue decreased significantly. However, in rats injected with hUCBMNCs for two times, no significant difference was discovered in MMP-7 levels and urinary albumin/creatinine ratio. Although expression of α-SMA and the percentage areas of collagen staining in tubulointerstitial tissues were ameliorated in rats injected with hUCBMNCs for two times, the effects were significantly weaker than those in rats injected with hUCBMNCs for four times. Taken together, our study constructed a highly efficient, duplicable novel rat model of renal fibrosis after cisplatin-induced AKI. Multiple injections of hUCBMNCs may prevent renal interstitial fibrosis after cisplatin-induced AKI.

2.
J Hazard Mater ; : 121742, 2019 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-31796347

RESUMO

Oil shale semi-coke is the solid waste produced from the retorting process of oil shale, which may cause pollution to the environment without reasonable disposing. In this study, semi-coke was used as the bulking agent during composting to accelerate biodegradation of the organics as well as decrease the nitrogen loss. Results showed that the addition of semi-coke could accelerate biodegradation of the organics, with a raise in the organic matter loss from 44.99 % to 47.05 % compared with the control. Furthermore, the nitrogen loss significantly decreased from 40.00%-14.70 % in the treatment added with semi-coke due to less emission of NH3 and much more transformation of NH4+-N to NO3--N by nitrification, which could be explained by the increasing abundance of ammonia-oxidizing bacteria and archaea at the late composting stage and drastic shift of the microbial community like Chloroflexi, Firmicutes and Actinobacteria. After the composting cycle, the maturity of the produced compost was elevated greatly in the treatments amended with semi-coke. The result of PAHs detection suggested that there were low PAHs content in the raw oil shale semi-coke and they could be removed effectively to within the range for land application by composting especially when the surfactant was added.

3.
Cancer Invest ; : 1-12, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31797701

RESUMO

Purpose: The function of long noncoding RNAs (lncRNA) in breast cancer metastasis remains largely unknown. In this work, the role of HOXC-AS3 in breast cancer progression was investigated.Methods: By using Cancer Genome Atlas (TCGA) Database, we investigated the expression of HOXC-AS3 in breast cancer and explored the association between HOXC-AS3 expression and prognosis. Then, we studied the biological function of HOXC-AS3 in cell migration and invasion both in vitro and in vivo. Furthermore, the target miRNA of HOXC-AS3, and the target mRNA of miR-3922-5p were proved.Results: HOXC-AS3 is aberrantly overexpressed in breast cancers especially the HER2+ type. Moreover, high expression of HOXC-AS3 has a relationship with poor clinical outcomes of breast cancer. In addition, HOXC-AS3 regulates cell Invasion and migration both in vitro and in vivo. Our results demonstrated that miR-3922-5p was a direct target of HOXC-AS3, and PPP1R1A was a target of miR-3922-5p in breast cancer.Conclusions: The novel lncRNA HOXC-AS3 acts as a miR-3922-5p sponge to upregulate PPP1R1A protein expression, and thus results in promoting breast cancer metastasis. HOXC-AS3 could be a novel therapeutic target for breast cancer therapeutics.

4.
Artigo em Inglês | MEDLINE | ID: mdl-31789936

RESUMO

OBJECTIVES: Intestinal neuronal dysplasia (IND) is a common malformation of the enteric nervous system. Diagnosis requires a full-thickness colonic specimen and an experienced pathologist, emphasizing the need for noninvasive analytical methods. Recently, the methylation level of the Sox10 promoter has been found to be critical for enteric nervous system development. However, whether it can be used for diagnostic purposes in IND is unclear. METHODS: Blood and colon specimens were collected from 32 patients with IND, 60 patients with Hirschsprung disease (HD), and 60 controls. Sox10 promoter methylation in the blood and the Sox10 expression level in the colon were determined, and their correlation was analyzed. The diagnostic efficacy of blood Sox10 promoter methylation was analyzed by receiver operating characteristic curve. RESULTS: The blood level of Sox10 promoter methylation at the 32nd locus was 100% (90%-100%; 95% confidence interval [CI], 92.29%-96.37%) in control, 90% (80%-90%; 95% CI, 82.84%-87.83%) in HD, and 60% (50%-80%; 95% CI, 57.12%-69.76%) in IND specimens. Sox10 promoter methylation in the peripheral blood was negatively correlated with Sox10 expression in the colon, which was low in control, moderate in HD, and high in IND specimens (r = -0.89). The area under the curve of Sox10 promoter methylation in the diagnosis of IND was 0.94 (95% CI, 0.874-1.000, P = 0.000), with a cutoff value of 85% (sensitivity, 90.6%; specificity, 95.0%). By applying a cutoff value of 65%, promoter methylation was more indicative of IND than HD. DISCUSSION: The analysis of Sox10 promoter methylation in the peripheral blood can be used as a noninvasive method for IND diagnosis.

5.
Artigo em Inglês | MEDLINE | ID: mdl-31792194

RESUMO

Circadian clocks usually run with a period close to 24 h, but are also plastic and can be entrained by external environmental conditions and internal physiological cues. Two key nutrient metabolites, glucose and vitamin B3 (nicotinamide), can influence the circadian period in both mammals and plants; however, the underlying molecular mechanism is still largely unclear. We reveal that the target of rapamycin (TOR) kinase, a conserved central growth regulator, is essential for glucose- and nicotinamide-mediated control of the circadian period in Arabidopsis Nicotinamide affects the cytosolic adenosine triphosphate concentration, and blocks the effect of glucose-TOR energy signaling on period length adjustment, meristem activation, and root growth. Together, our results uncover a missing link between cellular metabolites, energy status, and circadian period adjustment, and identify TOR kinase as an essential energy sensor to coordinate circadian clock and plant growth.

6.
Orthop Surg ; 2019 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-31755245

RESUMO

The iASSIST navigation system is a handheld accelerometer-based navigation system that has been applied in clinical practice in recent five years. This meta-analysis aimed to compare the radiographic and clinical outcomes of iASSIST navigation with conventional surgical techniques for patients undergoing total knee arthroplasty (TKA) and to compare the surgery time between an iASSIST group and a conventional treatment group. This systematic review and meta-analysis included all comparative prospective and retrospective studies published in Pubmed, Embase, the Cochrane Central Register of Controlled Trials, the Web of Science and the CNKI databases over the past 20 years. Inclusion criteria were studies that compared the iASSIST navigation system with conventional TKA. The primary outcomes were mechanical axis (MA) and outliers, which means postoperative MA varus or valgus of more than 3°. Secondary outcomes were coronal femoral angle (CFA) and coronal tibial angle (CTA). Knee Society Score (KSS) was used to evaluate functional outcome. The Newcastle-Ottawa Scale (NOS) was used to assess the methodological quality of included studies. Eight studies involving 558 knees were included in this meta-analysis. Of these, 275 patients used the iASSIST navigation system and 283 used conventional surgical techniques. A total of 5 studies were considered high quality and the other 3 were considered to be of moderate quality. The occurrence of malalignment of >3° in the iASSIST group was 13.3%, compared with 29.04% in the conventional group. Postoperative MA of the iASSIST group was significantly better than that of the conventional group (I2 = 19%, OR = -0.92, 95% CI = -1.09 to -0.75, P < 0.00001). The iASSIST navigation system provided significantly increased accuracy in the coronal femoral angle (I2 = 79%, OR = -0.88, 95% CI = -1.21 to -0.54, P < 0.00001) and the coronal tibial angle (I2 = 34%, OR = 0.39, 95% CI = -0.48 to -0.30, P < 0.00001) compared with conventional techniques. However, the duration of surgery using the iASSIST procedure was longer and there was no significant difference in the short-term KSS in the iASSIST group compared with the conventional group. We found that when pooling the data of included studies, the number of outliers was fewer in the iASSIST group, and compared with conventional TKA techniques, the iASSIST system significantly improved the accuracy of lower limb alignment but the duration of surgery was prolonged in addition to there being no apparent advantage in terms of short-term functional score.

7.
Lung Cancer ; 139: 118-123, 2019 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-31775086

RESUMO

OBJECTIVES: The 2015 World Health Organization classification defines pulmonary large-cell neuroendocrine carcinoma (LCNEC) as a high-grade neuroendocrine carcinoma. However, the clinical characteristics and prognostic factors of pure LCNEC and combined LCNEC remain unclear. Hence, we performed a multi-center retrospective study to compare the clinical outcomes of pure versus combined LCNEC. MATERIALS AND METHODS: Data from 381 patients with pulmonary LCNEC admitted to 17 Chinese institutes between 2009 and 2016 were collected retrospectively. Clinical characteristics and prognosis were analyzed among patients receiving adjuvant (adjuvant group; n = 56) and first-line (first-line group; n = 146) chemotherapy, as well as among patients receiving small cell lung cancer (SCLC) and non-SCLC (NSCLC) chemotherapy regimens. The Kaplan-Meier method and multivariable Cox regression were used to identify clinicopathological variables that might influence patient outcomes. RESULTS: Expression levels of neuroendocrine markers (synaptophysin, chromogranin-A, CD56) were associated with patients' prognosis in the total study cohort. In the adjuvant group, median disease-free survival was non-significantly longer for SCLC-based regimens than for NSCLC-based regimens (P = 0.112). In the first-line group, median progression-free survival was significantly longer for SCLC-based regimens than for NSCLC-based regimens (11.5 vs. 7.2 months, P = 0.003). Among patients with combined LCNEC, adenocarcinoma was the most common combined component, accounting for 70.0 % of cases. Additionally, median overall survival was non-significantly shorter for combined LCNEC than for pure LCNEC (P = 0.083). CONCLUSION: The SCLC regimen is a more effective choice, as either first-line or adjuvant chemotherapy, when compared to the NSCLC regimen for LCNEC treatment. Further studies are needed to clarify the survival differences between patients with pure-, and combined LCNEC.

8.
Cancer Gene Ther ; 2019 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-31676843

RESUMO

Liposomes are one of the most widely investigated carriers for CRISPR/Cas9 delivery. The surface properties of liposomal carriers, including the surface charge, PEGylation, and ligand modification can significantly affect the gene silencing efficiency. Three barriers of systemic CRISPR/Cas9 delivery (long blood circulation, efficient tumor penetration, and efficient cellular uptake/endosomal escape) are analyzed on liposomal carriers with different surface charges, PEGylations, and ligand modifications. Cationic formulations dominate CRISPR/Cas9 delivery and neutral formulations also have good performance while anionic formulations are generally not proper for CRISPR/Cas9 delivery. The PEG dilemma (prolonged blood circulation vs. reduced cellular uptake/endosomal escape) and the side effect of repeated PEGylated formulation (accelerated blood clearance) were discussed. Effects of ligand modification on cationic and neutral formulations were analyzed. Finally, we summarized the achievements in liposomal CRISPR/Cas9 delivery, outlined existing problems, and provided some future perspectives. Liposomes are one of the most widely investigated carriers for CRISPR/Cas9 delivery. The surface properties of liposomal carriers, including the surface charge, PEGylation, and ligand modification can significantly affect the gene silencing efficiency. Three barriers of systemic siRNA delivery (long blood circulation, efficient tumor penetration, and efficient cellular uptake/endosomal escape) are analyzed on liposomal carriers with different surface charges, PEGylations, and ligand modifications. Cationic formulations dominate CRISPR/Cas9 delivery and neutral formulations also have good performance while anionic formulations are generally not proper for CRISPR/Cas9 delivery. The PEG dilemma (prolonged blood circulation vs. reduced cellular uptake/endosomal escape) and the side effect of repeated PEGylated formulation (accelerated blood clearance) were discussed. Effects of ligand modification on cationic and neutral formulations were analyzed. Finally, we summarized the achievements in liposomal CRISPR/Cas9 delivery, outlined existing problems, and provided some future perspectives.

9.
Cell Cycle ; 18(24): 3472-3490, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31713447

RESUMO

Protein kinase CK2 alpha (CK2α) is involved in the development of multiple malignancies. Overexpression of Y-box binding protein 1 (YBX1) is related to tumor proliferation, drug resistance, and poor prognosis. Studies have demonstrated that both CK2 and YBX1 could regulate the PI3K/AKT pathway. In addition, we predicted that CK2 might be the upstream kinase of YBX1 through the Human Protein Reference Database (HPRD). Herein, we hypothesize that CK2 may interact with YBX1 and they regulate the PI3K/AKT signaling pathway together. Expressions of CK2α and YBX1 in cancer cell lines were evaluated by immunoblotting. The results showed that CK2α could regulate the expression of YBX1 at the transcriptional level, which is dependent on its enzymatic activity. Synergistic effects of PI3K/AKT pathway inactivation could be observed through combined inhibition of CK2α and YBX1, and YBX1 was required for CK2α-induced PI3K/AKT pathway activation. Further results demonstrated that CK2α could interact with YBX1 and PI3K/AKT antagonist decreased cell resistance to doxorubicin induced by co-activation of CK2α and YBX1. These results indicated that combined inhibition of CK2α and YBX1 showed synergistic effects in inactivating the PI3K/AKT signaling pathway and may be one of the mechanisms involved in tumor growth and migration.

10.
FEBS Lett ; 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31675429

RESUMO

Cyclobutane pyrimidine dimers (CPD), as a common DNA damage caused by UV radiation, often lead to skin cancer. Here, we identified a photolyase from the alga Arthrospira platensis (designated as Ap-phr), which has been regarded as a safe organism for humans for centuries, that can efficiently repair CPD lesions in ssDNA and dsDNA in vitro. The 1.6 Å resolution crystal structure of Ap-phr revealed that it possesses a unique methenyltetrahydrofolate chromophore-binding pattern with high energy transfer efficiency. Our study of Ap-phr highlights its potential use in cosmetic, industrial and aesthetic medicine applications.

11.
Chin Med J (Engl) ; 132(22): 2737-2744, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31725458

RESUMO

OBJECTIVE: To review the diagnosis of chronic wound biofilms and discuss current treatment approaches. DATA SOURCES: Articles included in this review were obtained from the following databases: Wanfang, China National Knowledge Infrastructure, PubMed, and the Web of Science. We focused on research published before August 2019 with keywords including chronic wound, biofilm, bacterial biofilms, and chronic wound infection. STUDY SELECTION: Relevant articles were selected by carefully reading the titles and abstracts. Further, different diagnosis and clinical treatment methods for chronic wound biofilm were compared and summarized from the selected published articles. RESULTS: Recent guidelines on medical biofilms stated that approaches such as the use of scanning electron microscopy and confocal laser scanning microscopy are the most reliable types of diagnostic techniques. Further, therapeutic strategies include debridement, negative pressure wound therapy, ultrasound, antibiotic, silver-containing dressing, hyperbaric oxygen therapy, and others. CONCLUSION: This review provides the identification and management of biofilms, and it can be used as a tool by clinicians for a better understanding of biofilms and translating research to develop best clinical practices.

12.
EMBO J ; : e102406, 2019 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-31782549

RESUMO

The Hippo pathway, which plays a critical role in organ size control and cancer, features numerous WW domain-based protein-protein interactions. However, ~100 WW domains and 2,000 PY motif-containing peptide ligands are found in the human proteome, raising a "WW-PY" binding specificity issue in the Hippo pathway. In this study, we have established the WW domain binding specificity for Hippo pathway components and uncovered a unique amino acid sequence required for it. By using this criterion, we have identified a WW domain-containing protein, STXBP4, as a negative regulator of YAP. Mechanistically, STXBP4 assembles a protein complex comprising α-catenin and a group of Hippo PY motif-containing components/regulators to inhibit YAP, a process that is regulated by actin cytoskeleton tension. Interestingly, STXBP4 is a potential tumor suppressor for human kidney cancer, whose downregulation is correlated with YAP activation in clear cell renal cell carcinoma. Taken together, our study not only elucidates the WW domain binding specificity for the Hippo pathway, but also reveals STXBP4 as a player in actin cytoskeleton tension-mediated Hippo pathway regulation.

13.
FEBS J ; 2019 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-31725950

RESUMO

The eubacterial ß sliding clamp (DnaN) plays a crucial role in DNA metabolism through direct interactions with DNA, polymerases, and a variety of protein factors. A canonical protein-DnaN interaction has been identified in Escherichia coli and some other species, during which protein partners are tethered into the conserved canonical hydrophobic crevice of DnaN via the consensus ß-binding motif. Caulobacter crescentus is an excellent research model for use in the investigation of DNA replication and cell-cycle regulation due to its unique asymmetric cell division pattern with restricted replication initiation; however, little is known about the specific features of C. crescentus DnaN (CcDnaN). Here, we report a significant divergence in the association of CcDnaN with proteins based on docking analysis and crystal structures that show that the ß-binding motifs of its protein partners bind a novel pocket instead of the canonical site. Pull-down and isothermal titration calorimetry results revealed that mutations within the novel pocket disrupt protein-CcDnaN interactions. It was also shown by replication and regulatory inactivation of DnaA assays that mediation of protein interaction by the novel pocket is closely related to the performance of CcDnaN during replication and the DnaN-mediated regulation process. Moreover, assessments of clamp competition showed that DNA does not compete with protein partners when binding to the novel pocket. Overall, our structural and biochemical analyses provide strong evidence that CcDnaN employs a noncanonical protein association pattern.

14.
Artigo em Inglês | MEDLINE | ID: mdl-31782597

RESUMO

Inspired by natural anti-inflammatory benzoxepanes, a series of new benzoxepane derivatives were designed and synthesized, and 10i emerged as the most effective compound in vitro with low toxicity. Further in vivo evaluation revealed that 10i could both ameliorate sickness behaviour through anti-inflammation in LPS-induced neuroinflammatory mice model and ameliorate cerebral ischemic injury through anti-neuroinflammation in rats subjected to transient middle cerebral artery occlusion. Encouraged by the promising results, target fishing of 10i was then performed by design of photo-affinity probes, followed by photo-cross-linking, click reaction, and LC-MS/MS, leading to identification of PKM2 as a key target protein responsible for anti-inflammatory effect of 10i. Furthermore, 10i exhibited anti-neuroinflammatory effect in vitro and in vivo via inhibiting PKM2-mediated glycolysis and NLRP3 activation, indicating PKM2 as a novel target for neuroinflammation and its related brain disorders. In addition, 10i encompassed much more safety profile compared to shikonin, a reported PKM2 inhibitor, suggesting that 10i could be used as a lead compound targeting PKM2 for the treatment of inflammation-related diseases such as ischemic stroke.

15.
Biomed Res Int ; 2019: 2645926, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31687382

RESUMO

Background. The population of patients with acute pancreatitis treated by the staff at our department of gastroenterology includes those with mild and self-limited disease ranging to those with severe and fatal disease. Early diagnosis and accurate prediction of the severity and outcome of this disease, which is commonly seen by our department, is important for a successful outcome. Metabolic comorbidities (e.g., diabetes mellitus, fatty liver, obesity, and metabolic syndrome) are relevant to the severity and progression of many diseases. The objective of this review was to examine clinical relationships between metabolic comorbidities and occurrence, severity, and outcome of acute pancreatitis.

16.
Artigo em Inglês | MEDLINE | ID: mdl-31699666

RESUMO

BACKGROUND: Lower limb chronic venous disease (CVD), resulting from iliac vein compression syndrome (IVCS), manifests as a series of symptoms ranging from varicose veins to venous ulcerations. Stent implantation has been considered an effective treatment method; however, the management of CVD has rarely been reported. In the present study, we evaluated the treatment and outcomes of patients with CVD. METHODS: We performed a retrospective cohort study of patients with severe iliac vein stenosis with lower limb CVD. The patients were divided into two groups: group 1 had received stenting alone (n = 42), and group 2 had received stenting and high ligation/endovenous laser treatment (n = 29). We evaluated the clinical outcomes using the Venous Clinical Severity Score and visual analog scale, and assessed the quality of life (QoL) using the Chronic Venous Disease QoL questionnaire at a median follow-up point of 15 months (range, 6-25 months). RESULTS: In our cohort, the prevalence rate of nonthrombotic IVCS (NIVCS) was 11.7% (98 of 838 patients). The technical success rate was 100%, without severe complications. During the study period, three group 1 patients and two group 2 patients were lost to follow-up. The overall patency rate in the patients with NIVCS during a mean follow-up period of 15.0 months (range, 6-25 months) was 94.4%. For patients with a Clinical, Etiology, Anatomy, Pathophysiology (CEAP) clinical class of <4, all parameters showed similar improvements in the two groups, except for the disappearance of varicose veins. However, in patients with a CEAP clinical class of ≥4, the combination therapy significantly improved their QoL. The Venous Clinical Severity Score reduction was 4.64 ± 1.72 in group 1 and 11.89 ± 1.82 in group 2 (P < .01). Pain, scored using the visual analog scale, demonstrated a decrease from 4.41 to 2.52 (P < .05) in group 1 and 4.71 to 0.53 (P < .01) in group 2. The relief rate of stasis dermatitis in groups 1 and 2 was 26.9% and 90.5%, respectively (P < .05), and the venous ulceration healing rate was 16.7% and 87.5%, respectively (P < .05). CONCLUSIONS: The prevalence of NIVCS should not be overlooked. The proposed combination treatment is an effective therapeutic strategy for patients with NIVCS and advanced CVD (CEAP clinical class, ≥4) during short-term follow-up.

17.
Artigo em Inglês | MEDLINE | ID: mdl-31702393

RESUMO

Background: EGFR-AS1 has been characterized as an oncogenic lncRNA in many types of cancers, while its roles in esophageal squamous cell carcinoma (ESCC) are unknown. Results: Their data showed that EGFR-AS1 and ROCK1 were upregulated in ESCC and positively correlated. Survival analysis showed that high EGFR-AS1 and ROCK1 expression levels predicted poor survival. In ESCC cells, EGFR-AS1 overexpression led to upregulated ROCK1, while miR-145 overexpression led to downregulated ROCK1 and reduced effects of EGFR-AS1 overexpression. Bioinformatics analysis showed that miR-145 may bind EGFR-AS1, while overexpression of EGFR-AS1 and miR-145 did not significantly affect each other. In esophageal squamous cell carcinoma (ESCC) cells, EGFR-AS1 and ROCK1 overexpression mediated the increased rates of ECSS cell invasion and migration. Overexpression of miR-145 played an opposite role and attenuated the effects of EGFR-AS1 overexpression. Conclusion: Therefore, EGFR-AS1 may upregulate ROCK1 by sponging miR-145 to promote ESCC cell invasion and migration.

18.
Bioconjug Chem ; 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31714753

RESUMO

Single-molecule force spectroscopy is a powerful tool to directly measure protein-protein interactions (PPI). The high specificity and precision of PPI measurements made it possible to reveal detailed mechanisms of intermolecular interactions. However, protein aggregation due to specific or nonspecific interactions is among the most challenging problems in PPI examination. Here, we propose a strategy of a parallel DNA circuit to probe PPI using single-molecule magnetic tweezers. In contrast to PPI examination using atomic force microscopy, microspheres as probes used in magnetic tweezers avoided the single-probe issue of a cantilever. Negatively charged DNA as a linker circumvented the severe aggregation in the PPI construct with a protein linker. The unnatural amino acid encoded in proteins of interest expanded the choices of biorthogonal conjugation. We demonstrated how to apply our strategy to probe the PPI between the PHD3-Bromo and the histone H3 methylated at K4, a critical epigenetic event in leukemia development. We found a rupture force of 12 pN for breaking the PPI, which is much higher than that required to peel DNA off from a nucleosome, 3 pN. We expect that our methods will make PPI measurements of mechanics and kinetics with great precision, facilitating PPI-related research, e.g., PPI-targeted drug discovery.

19.
Med Sci Monit ; 25: 8172-8180, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31670317

RESUMO

BACKGROUND Baicalin, one of the main bioactive components extracted from the traditional Chinese medicine baical Skullcap root, has an anti-tumor activity which had been studied in several cancers. However, its role in human mesothelioma remains unknown. In this study, we investigated the anti-tumor mechanisms of baicalin in the mesothelioma cell line MESO924. MATERIAL AND METHODS Effects of baicalin on mesothelioma were assessed by measuring cell viability, apoptosis, migration, invasion, inactivation of signaling intermediates, and cell-cycle alterations. RESULTS Baicalin inhibited the proliferation, migration, and invasion of human mesothelioma cells and increased their apoptosis, all in a dose-dependent manner. Specifically, baicalin decreased the expression of p-EGFR, p-AKT, p-MAPK, p-S6, Bcl-2, and VEGF and increased the expression of Bax in mesothelioma cells. The suppressed mesothelioma cellular proliferation is due to the arrest of the S cell cycle by baicalin. Inhibition of the PI3K/AKT/mTOR signaling pathway by a PI3K/AKT/mTOR inhibitor augmented the anti-proliferation effects induced by baicalin. In addition, baicalin increased the sensitivity of MESO924 to the chemotherapeutic drugs doxorubicin, cisplatin, and pemetrexed. CONCLUSIONS These results highlight the roles of baicalin in inhibiting cell growth, migration, and invasion of mesothelioma cells while increasing apoptosis and sensitizing cells to chemotherapeutic agents through the PI3K/AKT/mTOR signaling pathway, which indicates that baicalin could be a useful drug for mesothelioma therapy.

20.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(5): 581-588, 2019 Oct 30.
Artigo em Chinês | MEDLINE | ID: mdl-31699186

RESUMO

Objective To evaluate the effect of miR-145 on migration and invasion of ovarian cancer cells.Methods The effect of miR-145 overexpression on the expression levels of miR-145 and zeb-2 were detected with qRT-PCR and Western blotting.The changes of in vitro migration and invasion were examined using Transwell assay.Target genes of miR-145 were predicted by bioinformatics software.Dual-luciferase reporter assay were used to verify zeb-2 as a direct target of miR-145.zeb-2 siRNA was transiently transfected in SKOV3 and 3AO cells,Transwell was used to examine in vitro migration and invasion abilities.Results The migration and proliferation of SKOV3(t=10.752,P=0.000;t=5.617,P=0.005)and 3AO cells(t=10.111,P=0.001;t=21.746,P=0.000)decreased significantly after overexpression of miR-145.The results of dual-luciferase reporter assay showed that the relative luciferase activity of co-transfected miR-145 mimic and WT 3'UTR expression vectors was significantly lower than that of co-transfected mimic control and WT 3'UTR expression vectors(SKOV3:t=4.572,P=0.010;3AO:t=3.528,P=0.024).There was no significant difference in relative luciferase activity between co-transfected miR-145 mimic/MUT 3'UTR expression vector cells and co-transfected mimic control/MUT 3'UTR expression vector cells(SKOV3:t=0.227,P=0.831;3AO:t=0.040,P=0.970).Real-time quantitative PCR showed that the zeb-2 expressions in SKOV3(t=1.490,P=0.211)and 3AO cells(t=0.114,P=0.914)were not significantly different from negative control after 48 h of miR-145 overexpression.Western blot analysis showed that the expression of zeb-2 protein in SKOV3(t=3.769,P=0.020)and 3AO cells(t=4.452,P=0.011)decreased significantly compared with negative control after 72 h of miR-145 overexpression.Seventy-two hours after transfection of zeb-2 siRNA,Western blotting showed that the expression of zeb-2 protein in SKOV3(t=4.660,P=0.010)and 3AO cells(t=4.594,P=0.010)was significantly down-regulated.Transwell assay showed that the migration and invasion abilities of SKOV3(t=18.655,P=0.000;t=18.026,P=0.000)and 3AO cells(t=5.500,P=0.005;t=8.780,P=0.001)were significantly decreased.Conclusion miR-145 may inhibit the migration and invasion of ovarian cancer cells by targeting zeb-2.


Assuntos
Movimento Celular , MicroRNAs/genética , Invasividade Neoplásica , Neoplasias Ovarianas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Humanos , Homeobox 2 de Ligação a E-box com Dedos de Zinco/genética
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