Structure-based design and synthesis of anti-fibrotic compounds derived from para-positioned 3,4,5-trisubstituted benzene.

Liver fibrosis is an abnormal wound-healing response to liver injuries. It can lead to liver cirrhosis, and even liver cancer and liver failure. There is a lack of treatment for liver fibrosis and it is of great importance to develop anti-fibrotic drugs. A pivotal event in the process of developing liver fibrosis is the activation of hepatic stellate cells (HSCs), in which the nuclear receptor Nur77 plays a crucial role. This study aimed to develop novel anti-fibrotic agents with Nur77 as the drug target by modifying the structure of THPN, a Nur77-binding and anti-melanoma compound. Specifically, a series of para-positioned 3,4,5-trisubstituted benzene ring compounds with long-chain backbone were generated and tested for anti-fibrotic activity. Among these compounds, compound A8 was with the most potent and Nur77-dependent inhibitory activity against TGF-ß1-induced activation of HSCs. In a crystal structure analysis, compound A8 bound Nur77 in a peg-in-hole mode as THPN did but adopted a different conformation that could interfere the Nur77 interaction with AKT, which was previous shown to be important for an anti-fibrotic activity. In a cell-based assay, compound A8 indeed impeded the interaction between Nur77 and AKT leading to the stabilization of Nur77 without the activation of AKT. In a mouse model, compound A8 effectively suppressed the activation of AKT signaling pathway and up-regulated the cellular level of Nur77 to attenuate the HSCs activation and ameliorate liver fibrosis with no significant toxic side effects. Collectively, this work demonstrated that Nur77-targeting compound A8 is a promising anti-fibrotic drug candidate.

Effect of miniscrew placement height on the distribution of biological forces produced by clear aligner for mandibular molar distalization / 口腔疾病防治

Objective@#To explore the effect of different miniscrew placement heights on the distribution of biological forces produced by clear aligner combined with intramaxillary traction for mandibular molar distalization, to identify the miniscrew location that is conducive to the protection of lower anterior tooth anchorage and to provide a reference that can be used when designing clinical treatments.@*Methods@#Mimics, GeomagicStudio 2017, SolidWorks 2016, and Ansys workbench were used to establish finite element analysis models and perform mechanical analysis under the following six working conditions: working condition 1 was the control group without miniscrews; working conditions 2 to 5 had miniscrew in the buccal bone cortex between the first and second molars of the lower jaw 10 mm, 7 mm, 4 mm, and 1 mm from the top of the alveolar crest, respectively; working condition 6 had the miniscrew in the center of the buccal tongue at the anterior edge of the ascending branch of the lower jaw 5 mm above the occlusal plane.@*Results@#On the sagittal axis, miniscrew anchorage caused distal displacement of all teeth. Compared to the control group, in the miniscrew group, the displacement of the anterior molars exceeded that of the second molars. On the vertical axis, the result in the control group was similar to backward bending; the results in the miniscrew groups resembled the effect of a lever, lowering the lateral incisors and canines and raising the central incisors and first premolars. On the coronal axis, the second premolars and the first molars showed lingual displacement in the control group, and only the premolars and first molars showed lingual displacementin the miniscrew groups. The canines were the teeth that were most strongly affected by the change in miniscrew placement height.@*Conclusion@#The higher the miniscrew position is, the stronger the protective effect on the anterior anchorage. According to the miniscrew placement height, the mandibular arch should be properly narrowed, the central incisors and first premolars should be lowered, and the lateral incisors and canines should be raised when designing clinical treatments.

Mandibular advancement with clear aligners and functional appliances in the treatment of skeletal Classâ ¡malocclusion: a systematic review and meta-analysis.

OBJECTIVES: This study aimed to conduct a meta-analysis of the efficacy of mandibular advance clear alig-ners with traditional functional appliances as the control group. METHODS: PubMed, Web of Science, Embase, Cochrane Library, China Biomedical Abstracts Database, China Knowledge Network Database, Wanfang Database, and Weipu Database were used in this study. The two groups of researchers screened the literature and extracted data based on the inclusion and exclusion criteria established by PICOS entries, and used the ROBINS-I scale for quality evaluation. Revman 5.4 and Stata 17.0 software were used for meta-analysis. RESULTS: Nine clinical controlled trials were included in this study with a total sample size of 283 cases. No significant difference was found in SNA, SNB, ANB, Go-Pog, U1-SN, Overjet, and other aspects between the invisible group and the traditional group in the treatment of skeletal class Ⅱ ma-locclusion patients; there was a 0.90° difference in mandibular plane angle between the two groups; the growth of the mandibular ramus (Co-Go) in the traditional group was 1.10 mm more than that in the invisible group; the lip inclination of the lower teeth in the invisible group was better controlled, 1.94° less than that in the control group. CONCLUSIONS: The invisible group can better control the lip inclination of the mandibular anterior teeth when guiding the mandible. Furthermore, the mandibular plane angle (MP-SN) can remain unchanged, but the growth of the mandibular ramus is not as good as the traditional group, and auxiliary measures should be taken to improve it in clinical practice.

Pathogenic mechanism of Vibrio vulnificus infection.

Vibrio vulnificus is a fatal, opportunistic human pathogen transmitted through the consumption of raw/undercooked seafood or direct contact. V. vulnificus infection progresses rapidly and has severe consequences; some cases may require amputation or result in death. Growing evidence suggests that V. vulnificus virulence factors and regulators play a large role in disease progression, involving host resistance, cellular damage, iron acquisition, virulence regulation and host immune responses. Its disease mechanism remains largely undefined. Further evaluation of pathogenic mechanisms is important for selecting appropriate measures to prevent and treat V. vulnificus infection. In this review, the possible pathogenesis of V. vulnificus infection is described to provide a reference for treatment and prevention.

Structural insights into the redox regulation of Oncomelania hupensis TRP14 and its potential role in the snail host response to parasite invasion.

The freshwater amphibious snail Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum, but little attention has been paid to the interaction between the two. In snails, the production of reactive oxygen species (ROS) by hemocytes has been shown to be vital for snail immune defense against schistosome infection. However, excessive ROS accumulation could lead to oxidative damage, requiring the antioxidant system for maintaining the cellular redox homeostasis. Previously we identified a thioredoxin-related protein of 14 kDa from O. hupensis (OhTRP14), and showed that it was involved in the scavenging of ROS in circulating hemocytes. Here, we confirmed that OhTRP14 plays a potential role in the snail host response to parasite challenge and determined the crystal structures of OhTRP14 in two different states (oxidized and transition state). The overall structure revealed a typical Trx fold and is similar to that of human TRP14 (hTRP14), but there were significant structural differences between the two states. Noticeably, there was a different pair of thiol groups from Cys30 and Cys44 in the transition state of OhTRP14, were with the similar separation of 2.9 Å as that (2.6 Å) between Cys41 and Cys44, but in a different orientation, suggesting that the Cys30 is likely to function as an important molecular switch involved in the oxidoreductase activity of OhTRP14. Comparative studies between OhTRP14 and hTRP14 by analyzing the surface characteristics, charge distribution and oxidoreductase activity toward insulin demonstrated they might have similar substrates. The results are expected to provide structural insights into the redox regulation of OhTRP14 and contribute to better understanding of TRP14 family. DATA DEPOSITION: The atomic coordinates of the structure and the structure factors were deposited in Protein Data Bank with PDB ID codes 7XQ3 and 7XPW.

[Diffusive Fluxes and Controls of N2O from Coastal Rivers in Tianjin City].

Rivers are an important emission source of greenhouse gases. To explore the spatial characteristics and influencing factors of N2O emission from the coastal rivers in Tianjin City, six rivers into the Bohai Sea from different land-use types were selected, and the N2O concentrations, saturation, and diffusive fluxes were measured using the headspace-gas chromatography method. The N2O concentration was in supersaturation, and the rivers were the source of atmospheric N2O. The average concentration, saturation, and diffusive fluxes of N2O were (23.85±15.20) nmol·L-1, (309.71±197.38)%, and (27.04±16.46) µmol·(m2·d)-1, with the ranges of 12.70-115.69 nmol·L-1, 164%-1502%, and 9.17-244.79 µmol·(m2·d)-1, respectively. The N2O concentrations and diffusive fluxes of the rivers presented great spatial heterogeneity, with the sewage river (Huangdipai River)>urban river (Haihe River main stream, Jiyun River)>suburban river (Duliujian River, Yongding Xinhe River)>agricultural river (Chaobai Xinhe River). The N2O concentration and diffusion fluxes were significantly correlated with salinity, nutrients, and carbon sources. NO3--N and TP contributed greatly to the diffusive flux differences. N2O production and emission greatly related to the nitrogen cycle process in the Tianjin River, and different forms of nitrogen variously contributed to N2O diffusive fluxes. The salinity gradient had the opposite effect on the N2O emission in urban rivers and drainage rivers. The N2O diffusive fluxes of the sewage river in Tianjin were significantly higher than that of other river types. In the future, due to the development of urbanization and the expansion of urban land, more management measures should focus on the hotspots such as the downstream of wastewater treatment plants of sewage rivers, the estuaries of urban rivers, and the residential gathering areas of suburban rivers to reduce N2O emission.

Overexpression of phosphatidylethanolamine-binding protein 4 (PEBP4) associates with recurrence of meningiomas.

BACKGROUND AND PURPOSE: Abnormal expression of phosphatidylethanolamine-binding protein 4 (PEBP4) has been identified in various types of malignant tumors. In the present study, we investigated the expression of PEBP4 in meningioma cases and examined whether PEBP4 expression was correlated with outcomes among these patients. MATERIALS AND METHODS: The expression levels of PEBP4 and Ki-67 in human meningioma tissues from 65 patients were evaluated by immunohistochemical staining. The correlation between PEBP4 immunoreactivity in meningioma samples and patients' clinical outcomes was examined using the Kruskal-Wallis correlation test. The prognostic value of PEBP4 expression in meningiomas patients also was investigated. RESULTS: Immunohistochemical analysis revealed up-regulated PEBP4 expression in both atypical and anaplastic meningiomas compared with classical meningiomas (13.38 ± 4.19% vs. 3.64 ± 2.04%, P < 0.001). PEBP4 immunoreactivity in meningioma samples was closely correlated with that for Ki-67 (Spearman r = 0.7922, P < 0.0001). PEBP4 expression was also associated with tumor differentiation grade and clinical recurrence (P < 0.05). Multivariate regression analysis showed with high PEBP4 expression was associated with a longer recurrence-free survival (hazard ratio=0.252, 95% confidence interval: 0.067-0.940, P = 0.040). CONCLUSION: PEBP4 may play an important role in the progression of meningioma, as high PEBP4 expression was associated with a higher pathological grade of meningioma. Moreover, PEBP4 expression may be a meaningful prognostic biomarker in meningioma.

Clinical efficacy and mechanism of mesenchymal stromal cells in treatment of COVID-19.

Coronavirus disease 2019 (COVID-19) is a highly infectious epidemic disease that has seriously affected human health worldwide. To date, however, there is still no definitive drug for the treatment of COVID-19. Cell-based therapies could represent a new breakthrough. Over the past several decades, mesenchymal stromal cells (MSCs) have proven to be ideal candidates for the treatment of many viral infectious diseases due to their immunomodulatory and tissue repair or regeneration promoting properties, and several relevant clinical trials for the treatment of COVID-19 have been registered internationally. Herein, we systematically summarize the clinical efficacy of MSCs in the treatment of COVID-19 based on published results, including mortality, time to symptom improvement, computed tomography (CT) imaging, cytokines, and safety, while elaborating on the possible mechanisms underpinning the effects of MSCs, to provide a reference for subsequent studies.

Structural and functional insights into macrophage migration inhibitory factor from Oncomelania hupensis, the intermediate host of Schistosoma japonicum.

Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum. As an irreplaceable prerequisite in the transmission and prevalence of schistosomiasis japonica, an in-depth study of this obligate host-parasite interaction can provide glimpse into the molecular events in the competition between schistosome infectivity and snail immune resistance. In previous studies, we identified a macrophage migration inhibitory factor (MIF) from O. hupensis (OhMIF), and showed that it was involved in the snail host immune response to the parasite S. japonicum. Here, we determined the crystal structure of OhMIF and revealed that there were distinct structural differences between the mammalian and O. hupensis MIFs. Noticeably, there was a projecting and structured C-terminus in OhMIF, which not only regulated the MIF's thermostability but was also critical in the activation of its tautomerase activity. Comparative studies between OhMIF and human MIF (hMIF) by analyzing the tautomerase activity, oxidoreductase activity, thermostability, interaction with the receptor CD74 and activation of the ERK signaling pathway demonstrated the functional differences between hMIF and OhMIF. Our data shed a species-specific light on structural, functional, and immunological characteristics of OhMIF and enrich the knowledge on the MIF family.

Knockdown of PEBP4 inhibits human glioma cell growth and invasive potential via ERK1/2 signaling pathway.

Phosphatidylethanolamine (PE)-binding protein 4 (PEBP4) is an antiapoptotic protein that is aberrantly expressed in various malignancies. We previously demonstrated that PEBP4 expression is dramatically induced in human gliomas and positively correlated with tumor grade and patient survival. However, the function of PEBP4 in human glioma development and underlying mechanisms remain largely unknown. By stable lentiviral vector-mediated silencing of PEBP4, we examined the effects of PEBP4 knockdown on the growth, apoptosis, and invasion of U251 and U373 human glioma cell lines using MTT, Transwell, colony formation, and flow cytometric assays. We examined the in vivo role of PEBP4 in tumor growth by inoculation of BALB/c nu/nu male mice with PEBP4-deficient U251 and U373 cells. The expression of cell cycle- and apoptosis-related proteins was analyzed by Western blotting and immunostaining. Knockdown of PEBP4 significantly reduced the proliferation and invasion of human glioma cells while inducing cell apoptosis by altering the expression of cell cycle- and apoptosis-related proteins. Mechanistically, PEBP4 knockdown led to activation of the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway, an effect that could be reversed by U0126, a selective inhibitor of MEK1/2 (upstream of ERK1/2), suggesting involvement of ERK1/2 signaling in the regulation of glioma development and progression by PEBP4. We identified PEBP4 as a novel regulator mediating human glioma cell proliferation, invasion, and apoptosis as well as tumor formation and growth. Therefore, PEBP4 may be a potential therapeutic target in human glioma treatment.

Phycocomes zhengii gen. nov., sp. nov., a marine bacterium of the family Rhodobacteraceae isolated from the phycosphere of Chlorella vulgaris.

A Gram-stain-negative, motile with flagellum, ovoid- or rod-shaped, aerobic bacterium, was isolated from the phycosphere of the microalga Chlorellavulgaris and designated as strain LMIT002T. The bacterium formed white, circular and smooth colonies on marine agar 2216 after 48 h incubation at 25 °C. On the basis of 16S rRNA gene sequence analysis, strain LMIT002T was found to be affiliated with the family Rhodobacteraceae of the order Rhodobacterales, and formed a distinct group. The 16S rRNA gene sequence similarities between strain LMIT002T and type strains in the family including Poseidonocella pacifica DSM 29316T, Roseobacter litoralis and Rhodovulum sulfidophilum were 95.1, 95.0 and 95.0 %, respectively. Strain LMIT002Tgrew optimally at 25 °C, pH 6.0 and in the presence of 2.0 % (w/v) NaCl. The genomic DNA G+C content was 67.0 mol% (the thermal denaturation method) or 66.9 % (genome sequencing). The sole respiratory quinone was ubiquinone-8, while the major fatty acids were summed feature 8 (C18 : 1 ω7с/C18 : 1 ω6с). The draft genome of strain LMIT002T was sequenced and annotated, with the results showing that it had a total size of 4 607 780 bp and comprised 4557 genes. Functional genes encoding the production of vitamin B12, indole-3-acetic acid and transform dimethylsulphoniopropionate were detected. Given its distinct genomic, morphological and physiological differences from previously described type strains, strain LMIT002T is proposed as a representative of a novel genus of the family Rhodobacteraceae, with the name Phycocomes zhengii gen. nov., sp. nov. The type strain is LMIT002T (=KCTC 62390T=CICC 24357T).

Structure-based optimization of salt-bridge network across the complex interface of PTPN4 PDZ domain with its peptide ligands in neuroglioma.

The PTP non-receptor type 4 (PTPN4) is an important regulator protein in learning, spatial memory and cerebellar synaptic plasticity; targeting the PDZ domain of PTPN4 has become as attractive therapeutic strategy for human neuroglioma. Here, we systematically examined the complex crystal structures of PTPN4 PDZ domain with its known peptide ligands; a number of charged amino acid residues were identified in these ligands and in the peptide-binding pocket of PDZ domain, which can constitute a complicated salt-bridge network across the complex interface. Molecular dynamics (MD) simulations, binding free energy calculations and continuum model analysis revealed that the electrostatic effect plays a predominant role in domain-peptide binding, while other noncovalent interactions such as hydrogen bonds and hydrophobic forces are also responsible for the binding. The computational findings were then used to guide structure-based optimization of the interfacial salt-bridge network. Consequently, five peptides were rationally designed using the high-affinity binder Cyto8-RETEV (RETEV-COOH) as template, including four single-point mutants (i.e. Cyto8-mtxe0: RETEE-COOH, Cyto8-mtxd-1: RETDV-COOH, Cyto8-mtxd-3: RDTEV-COOH and Cyto8-mtxk-4: KETEV-COOH) and one double-point mutant (i.e. Cyto8-mtxd-1k-4: KETDV-COOH). Binding assays confirmed that three (Cyto8-mtxd-1, Cyto8-mtxk-4 and Cyto8-mtxd-1k-4) out of the five designed peptides exhibit moderately or considerably increased affinity as compared to the native peptide Cyto8-RETEV.

Highly water-soluble platinum(II) complexes as GLUT substrates for targeted therapy: improved anticancer efficacy and transporter-mediated cytotoxic properties.

Glucose-conjugated malonato-platinum(II) complexes are designed and synthesized to target tumor-specific active transporters, namely, glucose transporters (GLUTs); the complexes exhibit much higher aqueous solubility by 150 times, improved potency in cytotoxicities by 10 times, and increased therapeutic index by over 30 fold compared to the newest generation of clinical drugs oxaliplatin.

Synthesis, characterization and in vitro anti-tumor activities of matrine derivatives.

Nineteen previously unreported matrine derivatives were synthesized and characterized using elemental analysis, infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and mass spectrometry. Target compounds 6a-6l and 7a-7c showed stronger inhibitory activities than matrine in the in vitro antitumor tests and inhibited the growth of the Hep7402, B16-F10, A549, and TW03 cell lines. In addition, compound 6i exhibited a potent antitumor activity similar to that of colchicine.

Synthesis of carbazolones and 3-acetylindoles via oxidative C-N bond formation through PIFA-mediated annulation of 2-aryl enaminones.

A series of carbazolone derivatives and 3-acetylindoles have been achieved via PIFA-mediated intramolecular cyclization of 2-aryl enaminones. This process allows the N-moiety on the side-chain to be annulated to the benzene ring via the metal-free oxidative aromatic C-N bond formation.

Pd-catalysed synthesis of isoquinolinones and analogues via C-H and N-H bonds double activation.

An atom economical synthesis of isoquinolinones and analogues via ligand-free Pd-catalysed C-H and N-H double activation has been developed. A series of isoquinolinones were obtained in good to excellent yields. Good regioselectivities were also observed during the activation reactions with unsymmetrical alkynes. A practical one-pot procedure for the preparation of N-H isoquinolinones is also described.

3D pharmacophore models for thromboxane A(2) receptor antagonists.

Thromboxane A(2) (TXA(2)) is an endogenous arachidonic acid derivative closely correlated to thrombosis and other cardiovascular diseases. The action of TXA(2) can be effectively inhibited with TXA(2) receptor antagonists (TXRAs). Previous studies have attempted to describe the interactions between the TXA(2) receptor and its ligands, but their conclusions are still controversial. In this study, ligand-based computational drug design is used as a new and effective way to investigate the structure-activity relationship of TXRAs. Three-dimensional pharmacophore models of TXRAs were built with HypoGenRefine and HipHop modules in CATALYST software. The optimal HypoGenRefine model was developed on the basis of 25 TXRAs. It consists of two hydrophobic groups, one aromatic ring, one hydrogen-bond acceptor and four excluded volumes. The optimal HipHop model contains two hydrophobic groups and two hydrogen-bond acceptors. These models describe the key structure-activity relationship of TXRAs, can predict their activities, and can thus be used to design novel antagonists.

3D-pharmacophore models for selective A2A and A2B adenosine receptor antagonists.

Three-dimensional pharmacophore models were generated for A2A and A2B adenosine receptors (ARs) based on highly selective A2A and A2B antagonists using the Catalyst program. The best pharmacophore model for selective A2A antagonists (Hypo-A2A) was obtained through a careful validation process. Four features contained in Hypo-A2A (one ring aromatic feature (R), one positively ionizable feature (P), one hydrogen bond acceptor lipid feature (L), and one hydrophobic feature (H)) seem to be essential for antagonists in terms of binding activity and A2A AR selectivity. The best pharmacophore model for selective A2B antagonists (Hypo-A2B) was elaborated by modifying the Catalyst common features (HipHop) hypotheses generated from the selective A2B antagonists training set. Hypo-A2B also consists of four features: one ring aromatic feature (R), one hydrophobic aliphatic feature (Z), and two hydrogen bond acceptor lipid features (L). All features play an important role in A2B AR binding affinity and are essential for A2B selectivity. Both A2A and A2B pharmacophore models have been validated toward a wide set of test molecules containing structurally diverse selective antagonists of all AR subtypes. They are capable of identifying correspondingly high potent antagonists and differentiating antagonists between subtypes. The results of our study will act as a valuable tool for retrieving structurally diverse compounds with desired biological activities and designing novel selective adenosine receptor ligands.

[Effect of photodynamic therapy combined with interstitial chemotherapy for gliomas].

OBJECTIVE: To investigate the therapeutic effect of photodynamic therapy (PDT) combined with interstitial chemo- therapy on gliomas after microsurgery. METHODS: Twenty-eight patients with glioma received microsurgery for glioma resection as much as possible, during which PDT was carried out and an Ommaya bursa implanted into the cavity left by the resected tumor. PDT was performed again 24 h after the operation and carmustine injected into the Ommaya bursa for interstitial chemotherapy at the different time after the operation. The follow-up study lasted for 2 years. RESULTS: Two of the 28 patients had relapse of glioma within 1 year after the operation, with the 1-year survival rate of 89.3% (25/28) and 2-year survival rate of 65.7% (17/28). CONCLUSIONS: PDT combined with interstitial chemotherapy following microsurgery provides a safe and effective treatment of glioma, which can inhibit glioma growth, decrease the recurrence rate, prolong the patients' survival and improve their quality of live.