Identifying ligands for the PHD1 finger of KDM5A through high-throughput screening.

PHD fingers are a type of chromatin reader that primarily recognize chromatin as a function of lysine methylation state. Dysregulated PHD fingers are implicated in various human diseases, including acute myeloid leukemia. Targeting PHD fingers with small molecules is considered challenging as their histone tail binding pockets are often shallow and surface-exposed. The KDM5A PHD1 finger regulates the catalytic activity of KDM5A, an epigenetic enzyme often misregulated in cancers. To identify ligands that disrupt the PHD1-histone peptide interaction, we conducted a high-throughput screen and validated hits by orthogonal methods. We further elucidated structure-activity relationships in two classes of compounds to identify features important for binding. Our investigation offers a starting point for further optimization of small molecule PHD1 ligands.

Identification and Optimization of Small Molecule Pyrazolopyrimidine TLR7 Agonists for Applications in Immuno-oncology.

Small molecule toll-like receptor (TLR) 7 agonists have gathered considerable interest as promising therapeutic agents for applications in cancer immunotherapy. Herein, we describe the development and optimization of a series of novel TLR7 agonists through systematic structure-activity relationship studies focusing on modification of the phenylpiperidine side chain. Additional refinement of ADME properties culminated in the discovery of compound 14, which displayed nanomolar reporter assay activity and favorable drug-like properties. Compound 14 demonstrated excellent in vivo pharmacokinetic/pharmacodynamic profiles and synergistic antitumor activity when administered in combination with aPD1 antibody, suggesting opportunities of employing 14 in immuno-oncology therapies with immune checkpoint blockade agents.

Nivalenol disrupts mitochondria functions during porcine oocyte meiotic maturation.

Oocyte maturation is important for fertility in mammals, since the quality of oocytes directly affects fertilization, embryo attachment and survival. Nivalenol is widely present in nature as a common toxin that contaminates grain and feed, and it has been reported to cause acute toxicity, immunotoxicity, reproductive toxicity and carcinogenic effects. In this study, we explored the impact of nivalenol on the porcine oocyte maturation and its possible mechanisms. The extrusion of the first polar body was significantly inhibited after incubating oocytes with nivalenol. Meanwhile, nivalenol exposure led to the abnormal distribution of mitochondria, aberrant calcium concentration and the reduction of membrane potential caused a significant decrease in the capacity of mitochondria to generate ATP. In addition, nivalenol induced oxidative stress, and the level of ROS was significantly increased in the nivalenol-treated group, which was confirmed by the perturbation of oxidative stress-related genes. We found that nivalenol-treated oocytes showed positive Annexin-V and γH2A.X signals, indicating the occurrence of apoptosis and DNA damage. In all, our data suggest that nivalenol disrupted porcine oocyte maturation through its effects on mitochondria-related oxidative stress, apoptosis and DNA damage.

Neuron-Specific Enolase and Hemoglobin as Risk Factors of Intraocular Metastasis in Patients with Renal Cell Carcinoma.

Renal cell carcinoma (RCC) appears to be a high risk of spread. This research investigated the correlation between a different range of clinical features and intraocular metastasis (IOM) in RCC patients and attempted to determine potential risk factors of RCC patients with IOM. In the study, there are a total of 351 patients with RCC that were recruited between May 1994 and May 2016. The differences between RCC patients with IOM and RCC patients with non-IOM (NIOM) were evaluated by the chi-squared test and Student t test. Binary logistic regression analysis was applied to determine risk factors. Finally, the value of diagnosis for RCC patients with IOM was assessed by receiver operating characteristic (ROC) curve analysis. Eighteen individuals were identified with IOM. There were no significant differences that were detected in alkaline phosphatase (AFP), carcinoembryonic antigen (CEA), alkaline phosphatase (ALP), cancer antigen 125 (CA-125), cancer antigen 153 (CA-153), cancer antigen 199 (CA-199), calcium, age, primary tumor site, and histopathological subtypes between the two groups. But there was a difference in terms of gender (P < 0.05). The IOM group exhibited significantly higher neuron-specific enolase (NSE) and lower hemoglobin (Hb) values compared to the NIOM group (P < 0.05, respectively). Binary logistic regression identified NSE and Hb as significant risk factors of IOM for RCC patient (P < 0.05 and P < 0.001, respectively). The ROC curve analysis indicated that the area under the curve (AUC) values of NSE and Hb were 0.694 and 0.749, while cut-off values were 49.5 ng/mL and 102.5 g/L, respectively. The sensitivity and specificity of NSE were 72.2% and 66.4%, respectively, while those of Hb were 72.2% and 74.2%, respectively. The result reveals that NSE and Hb represent promising significant risk factors of IOM for RCC patients. Notably, Hb is more reliable than NSE in distinguishing case of IOM from NIOM in patients with RCC.

Efficacy of Transosseous Tunnel Placement for Triple Endobutton Plate in Acromioclavicular Joint Reconstruction: A Three-Dimensional Printing Guide Design Technology.

OBJECTIVE: Explore an accurate transosseous tunnel drilling method based on three-dimensional (3D) printing technology for acromioclavicular joint reconstruction (ACD), design a guide design, and evaluate its accuracy. METHODS: Using Mimics software to reconstruct 100 cases of acromioclavicular joint computed tomography (CT) data. In design 2, the non-collinear tunnel is superimposed on the 3D model, and a virtual drilling is performed between the clavicle and the coracoid using a triple inner gusset. Then, in the Geomagic Studio software model, an elliptical plane is calculated and extracted as a guide design for precise drilling. Then put the design and the 3D shoulder model together for 3D printing. Ten lengths were measured, and the effects of the virtual model, the actual model, and the guide rail design were compared. RESULTS: We successfully compared 10 parameters of 3D virtual model and actual model. There was no significant difference between actual and virtual bone tunnels in 10 measurements (P > 0.05). CONCLUSIONS: The accuracy of ACD combined with 3D printing guidance design technology in the transosseous tunnel of adult shoulder is reliable.

Antibody prevalence and immunotherapy response in Chinese patients with epilepsy and encephalopathy scores for patients with different neuronal surface antibodies.

BACKGROUND: The scale assessment was helpful in predicting the presence of antibodies to autoimmune encephalitis. This study aimed to evaluate the application of antibody prevalence in Chinese patients with epilepsy and encephalopathy (APE2-CHN) and response to immunotherapy in Chinese patients with epilepsy and encephalopathy (RITE2-CHN) for patients with different neuronal surface antibodies. METHODS: A total of 1365 patients with epileptic seizures as the prominent feature in Xuanwu Hospital, Capital Medical University, from June 2016 to June 2020 were enrolled in our study. Of these, 915 patients with epilepsy of unknown etiology whose serum and/or cerebrospinal fluid samples were examined for autoimmune antibodies were selected. All patients were scored with antibody prevalence in patients with epilepsy and encephalopathy (APE2), response to immunotherapy with epilepsy and encephalopathy (RITE2), APE2-CHN, and RITE2-CHN scores. RESULTS: Of the 915 patients, 191 patients were positive for neural-surface specific antibodies (115 N-methyl-D-aspartate receptor (NMDAR) Ab, 47 leucine-rich glioma-inactivated protein 1 (LGI1) Ab, 8 contactin-associated protein 2 (CASPR2) Ab, 4 AMPA2R-Ab, and 11 GABAR-B-Ab; 3 CASPR2-Ab and LGI1-Ab, 2 NMDAR-Ab and CASPR2-Ab, and 1 NMDAR-Ab and myelin-oligodendrocyte glycoprotein [MOG] Ab). The sensitivity and specificity of APE2 ≥4 in predicting the presence of neural-surface specific antibodies in our study were 74.35% and 81.77%, respectively, and the sensitivity and specificity of APE2-CHN ≥4 were 75.92% and 84.53%, respectively. Eight cases had an APE2 score <4 and APE2-CHN score ≥5; all these patients had memory decline as the prominent manifestation. We divided the patients into six groups according to the different antibodies. APE2-CHN scores showed higher sensitivity for the prediction of NMDAR-Ab, but lower sensitivity for LGI1-Ab. A total of 187/191 (97.91%) patients received immunotherapy and 142/191 (74.35%) patients benefited from the treatments. The patients who were positive for LGI1-Ab with RITE2-CHN ≥8 responded well to immunotherapy. CONCLUSIONS: APE2-CHN had the highest value for predicting the positivity of NMDAR-Ab and RITE2-CHN evaluated the response of immunotherapy for anti-LGI1 encephalitis appropriately. However, RITE2 and RITE2-CHN do not appear to be good predictors of immunotherapy outcomes for patients with specific neuronal-surface antibodies and high APE2-CHN scores are often indicative of a poor response to immunotherapy.

Risk factors for intraocular metastasis of primary liver cancer in diabetic patients: Alpha-fetoprotein and cancer antigen 125.

BACKGROUND: In rare instances, primary liver cancer can be associated with intraocular metastasis (IOM). AIM: To investigate the correlation between a diverse range of clinical characteristics and IOM in diabetic patients with primary liver cancer, and to determine potential risk factors in predicting IOM. METHODS: We recruited a total of 722 diabetic patients with primary liver cancer. The differences between the IOM and non-intraocular metastasis (NIOM) groups in these patients were assessed using the chi-squared test and Student's t-test. Binary logistic regression analysis was subsequently used to determine risk factors. Finally, the diagnostic value of IOM in this cohort with primary liver cancer was analyzed by receiver operating characteristic (ROC) curve analysis. RESULTS: In all, 13 patients had IOM. There were no remarkable intergroup differences with respect to age, sex, histopathological sub-types, or blood biochemical parameters. However, the IOM group had significantly higher alpha-fetoprotein (AFP) and cancer antigen 125 (CA125) values than the NIOM group. Binary logistic regression identified AFP and CA125 to be significant risk factors for IOM in diabetic patients with primary liver cancer. ROC curve analysis showed that the area under the curve values for AFP and CA125 were 0.727 and 0.796, with the cut-off values of 994.20 ng/mL and 120.23 U/mL, respectively. The sensitivity and specificity for AFP were 92.3% and 59.9%, while those for CA125 were 84.6% and 70.1%, respectively. CONCLUSION: Elevated AFP and CA125 represent significant risk factors for IOM in diabetic patients with primary liver cancer.

The Relationship between Calcaneal Spur Type and Plantar Fasciitis in Chinese Population.

Plantar heel pain is a common disease with a high incidence in different races. It significantly reduced the quality of life of patients. However, the cause of PHP is still controversial and there were varieties of physiological factors associated with PHP. The most common pathological factor in the population was plantar fasciitis. Some existing research studies had found a correlation between calcaneal spurs and plantar fasciitis, and this study had found the correlation in Chinese population. It is invaluable not only to understand the relationship between different types of plantar calcaneal spurs and plantar fasciitis but also to identify the most appropriate treatment strategies. A total of 71 patients with calcaneal spurs were chosen from the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University. All 71 patients had completed X-rays and MRI scans; then, surgeons had removed their plantar calcaneal spurs. After surgery, all patients were followed up for 12 months; their prognosis was tested by the VAS and AOFAS scores. Type II (29, 40.8%) had the highest incidence in Chinese population, followed with type I (24, 33.8%) and type III (18, 25.4%). Preoperative VAS scores showed that type II (7.72 ± 1.10) was significantly higher than the other two types (P < 0.001). Postoperative VAS scores of type II were higher than those of type I and type III (P < 0.001). Postoperative AOFAS scores of type II were the lowest (P < 0.001). Researchers had proved that type II was more likely to cause PF.

Decarboxylative Couplings for Late-Stage Peptide Modifications.

The application of designer peptides in medicinal chemistry, chemical biology, and materials science has generated new interest in synthetic methods for the structural modification of amino acids. Strategies which facilitate the direct diversification of proteinogenic functional groups within unprotected peptide substrates are particularly attractive as they leverage modern solution- and solid-phase protocols-tools which are now both robust and routine-for the synthesis of native peptides. Accordingly, a recent approach to the decarboxylative functionalization of peptidic carboxylic acids, including aspartic/glutamic acid residues and α-carboxylic acids, has proven to be a promising new strategy for peptide modification. This synthetic method merges conventional strategies for the activation of carboxylic acids with transition metal-catalyzed cross-coupling chemistry to forge new C-C bonds for the late-stage introduction of valuable synthetic handles. This chapter details a step-by-step protocol for the activation and nickel-catalyzed decarboxylative alkylation of a simple peptide substrate to highlight the broad utility of this strategy for the synthesis of designer peptides.

Chachezhania antarctica gen. nov., sp. nov., a novel member of the family 'Rhodobacteraceae' isolated from Antarctic seawater.

A Gram-stain-negative, aerobic, non-flagellated, rod-shaped bacterium, designated strain SM1703T, was isolated from Antarctic seawater collected near the Chinese Antarctic Great Wall Station, King George Island, West Antarctica. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM1703T formed a distinct phylogenetic lineage within the family 'Rhodobacteraceae', sharing high 16S rRNA gene sequence similarity with Marivita litorea (95.5%). The strain grew at 10-37 °C (optimum, 25 °C) and with 0.5-13% (w/v) NaCl (optimum, 3-5%). The major cellular fatty acids were C19:0 cyclo ω8c, C18:1ω7c, C18:1 2-OH and C16:0 2-OH. The major polar lipids were phosphatidylglycerol, phosphatidylcholine, an unidentified aminolipid and an unidentified lipid. The genomic DNA G+C content of strain SM1703T was 64.6 mol%. Based on the results of the polyphasic characterization for strain SM1703T, it is classified as the representative of a novel species in a new genus of the family 'Rhodobacteraceae', for which the name Chachezhania antarctica gen. nov., sp. nov. is proposed. The type strain of Chachezhania antarctica is SM1703T (= MCCC 1K03470T = KCTC 62794T = CCTCC AB 2018351T).

MiR-23a targets RUNX2 and suppresses ginsenoside Rg1-induced angiogenesis in endothelial cells.

Rg1 is a predominant protopanaxatriol-type of ginsenoside found in Panax ginseng, and it has been shown to have anti-cancer effects in multiple types of cancer cells. However, Rg1 also induces the expression of proangiogenic factors, such as vascular endothelial growth factor (VEGF-A), in endothelial cells. Unfortunately, angiogenesis positively correlates with cancer development. In this study, we identified RUNX2 as a regulator of ginsenoside Rg1-induced angiogenesis for the first time. We found that RUNX2 was directly targeted and regulated by miR-23a. Additionally, miR-23a was shown to inhibit angiogenesis in both human umbilical vein endothelial cells (HUVECs) and in zebrafish. Furthermore, a decrease in RUNX2 expression resulted in translational repression of VEGF-A in HUVECs. Taken together, this study identified a MiR-23a/RUNX2/VEGF-A pathway in angiogenesis and shed light on the molecular mechanism of Rg1-induced angiogenesis. Thus, RUNX2 might be a potential therapeutic target in Rg1-mediated angiogenesis in cancer.

[Screening and inhibition of antagonistic endophytic bacteria isolated from soybean (Glycine max)nodules against Alternaria longipes.] / çƒŸè‰èµ¤æ˜Ÿç— èŒæ‹®æŠ—æ€§å¤§è±†æ ¹ç˜¤å† ç”ŸèŒçš„ç­›é€‰åŠæŠ‘åˆ¶ä½œç”¨.

Using Alternaria longipes as tested phytopathogen, endophytic bacteria isolated from soybean nodules were selected to study antagonistic effects by confrontation and metabolic liquid culture methods. The inhibited hyphae were determined by microscopic observation, and the screened strains were characterized by cell culture, physiological and biochemical tests, 16S rDNA sequencing, phylogenetic analysis and inoculation trials in greenhouse. The results indicated that the seven of the endophytes exerted more than 42% inhibitory effects after the first and the second screening. These strains belonged to genus Bacillus, Pseudomonas, Sinorhizobium and Stenotrophomonas, respectively. Microscopic observation showed that the affected hyphae ends of A. longipes appear deformity of coralline branch, spherical expansions and so on. Antagonistic experiments with metabolites showed that the inhibition of endophytic bacteria against pathogenic fungus played an effective role mainly by bacteria producing extracellular substances. Confrontation tests suggested that endophytic Bacillus rapidly produced biofilm to effectively hinder the growth and extension of pathogen hyphae. Inoculation experiments showed that the disease index of treatment group was significantly lowered compared with the control, suggesting it could be utilized as a biological control resource inhibiting tobacco brown spot.

MiR-200c suppresses TGF-ß signaling and counteracts trastuzumab resistance and metastasis by targeting ZNF217 and ZEB1 in breast cancer.

Resistance to trastuzumab and concomitantly distal metastasis are leading causes of mortality in HER2-positive breast cancers, the molecular basis of which remains largely unknown. Here, we generated trastuzumab-resistant breast cancer cells with increased tumorigenicity and invasiveness compared with parental cells, and observed robust epithelial-mesenchymal transition (EMT) and consistently elevated TGF-ß signaling in these cells. MiR-200c, which was the most significantly downregulated miRNA in trastuzumab-resistant cells, restored trastuzumab sensitivity and suppressed invasion of breast cancer cells by concurrently targeting ZNF217, a transcriptional activator of TGF-ß, and ZEB1, a known mediator of TGF-ß signaling. Given the reported backward inhibition of miR-200c by ZEB1, ZNF217 also exerts a feedback suppression of miR-200c via TGF-ß/ZEB1 signaling. Restoration of miR-200c, silencing of ZEB1 or ZNF217 or blockade of TGF-ß signaling increased trastuzumab sensitivity and suppressed invasiveness of breast cancer cells. Therefore, our study unraveled nested regulatory circuits of miR-200c/ZEB1 and miR-200c/ZNF217/TGF-ß/ZEB1 in synergistically promoting trastuzumab resistance and metastasis of breast cancer cells. These findings provide novel insights into the common role of EMT and related molecular machinery in mediating the malignant phenotypes of breast cancers.