磺酰脲类除草剂的三维药效团模型

磺酰脲类除草剂是七十年代末期[1，2]开发的一类超高效除草剂，杀草谱广，用量极低．它通过抑制植物体内乙酰乳酸合成酶（ALS，acetolactatesynthase）的活性[3]，破坏支链氨基酸缬氨酸、亮氨酸和异亮氨酸的合成，造成杂草的死亡．本文从三维角度出发，利用先进的三维分子设计软件APEX（ActivityPredictionExpertSystem）－3D[4]，建立破酸豚类除草剂药效团模型（biophoremodel），辅助设计、筛选活性化合物；再由药效团构造出模拟ALS酶活性位点（activesites）的空间结构，以该模拟结构为基点，以期设计出全新结构的ALS酶抑制剂的…     

γH2AX,an Accurate Marker That Analyzes UV Genotoxic Effects on Human Keratinocytes and on Human Skin

The phosphorylated form of histone H2AX, γH2AX, is a component of the DNA repair system. Most studies have focused on the role of γH2AX during cell transformation and human cancer, but little is known about its role in keratinocytes and the skin during UV irradiation. We analyzed the response to UV irradiation focusing on the phosphorylation of histone H2AX both in vitro, in keratinocyte cultures and in artificial epidermis, and then in vivo, in human skin. Acute UVB irradiation of human keratinocytes increased the phosphorylation of H2AX in a dose-dependent manner; two types of γH2AX response were observed either in vitro or in vivo. After a low nonapoptotic UVB irradiation, cells contained phosphorylated H2AX and arrested their cell cycle to repair the DNA damages. For a stronger and proapoptotic UVB irradiation, keratinocytes dramatically increased the phosphorylation of H2AX and committed apoptosis. Our results indicate that γH2AX constitutes a highly sensitive marker relevant for studying subapoptotic doses as well as proapoptotic doses of UVB in human skin.     

3D纸基模型表面亮度预测研究

纸基模型表面亮度感知是整体色貌感知的重要组成,亮度感知的准确性、快捷性直接影响到纸基模型整体外貌的视觉感知。本文通过对3D纸基模型表面亮度测量,实现3D纸基表面在不同角度、不同阶调时的亮度预测。首先,通过设计的纸模测量角度架,可以方便快捷地调整纸基表面角度,然后,确定测量角度取样间隔为5°,纸模表面角度为5°~85°,依次测量,最后,设计四色表面梯尺,青色、品色、黄色、黑色在不同纸基表面和不同阶调下,利用辐射亮度计对其表面亮度值进行测量。利用最小二乘法建立亮度与纸基表面角度、阶调之间的关系模型,并对模型优劣程度进行检验。结果表明,本预测模型可以快速准确地对任意角度3D纸基表面进行亮度预测,获取3D纸基模型表面的亮度。     

Combination of Oral Vitamin D3 with Photodynamic Therapy Enhances Tumor Cell Death in a Murine Model of Cutaneous Squamous Cell Carcinoma

Photodynamic therapy (PDT), in which 5‐ALA (a precursor for protoporphyrin IX, PpIX) is administered prior to exposure to light, is a nonscarring treatment for skin cancers. However, for deep tumors, ALA‐PDT is not always effective due to inadequate production of PpIX. We previously developed and reported a combination approach in which the active form of vitamin D₃ (calcitriol) is given systemically prior to PDT to improve PpIX accumulation and to enhance PDT‐induced tumor cell death; calcitriol, however, poses a risk of hypercalcemia. Here, we tested a possible strategy to circumvent the problem of hypercalcemia by substituting natural dietary vitamin D₃ (cholecalciferol; D₃) for calcitriol. Oral D₃ supplementation (10 days of a 10‐fold elevated D₃ diet) enhanced PpIX levels 3‐ to 4‐fold, and PDT‐mediated cell death 20‐fold, in subcutaneous A431 tumors. PpIX levels and cell viability in normal tissues were not affected. Hydroxylated metabolic forms of D₃ were only modestly elevated in serum, indicating minimal hypercalcemic risk. These results show that brief oral administration of cholecalciferol can serve as a safe neoadjuvant to ALA‐PDT. We suggest a clinical study, using oral vitamin D₃ prior to PDT, should be considered to evaluate this promising new approach to treating human skin cancer.     

Metabonomics and Molecular Biology-based Effects of Sugemule-3 in an Isoproterenol-induced Cardiovascular Disease Rat Model

Cardiovascular disease(CVD) is a common and serious disease in the elderly, which has characteristically high prevalence, disability, and mortality rates. However, the etiology of CVD is still not very clear. The traditional Mongolian medicine Sugemule-3(SM) is usually used for the treatment of CVD and exhibits a good curative effect. In this study, a serum metabolite profile analysis was used to identify potential biomarkers associated with isoproterenol(ISO)-induced CVD and investigate the mechanism of the action of SM. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used for the metabono-mics analysis. Principal component analysis(PCA) was used to process the acquired data to differentiate the results of the control, CVD, and SM treatment groups. Orthogonal partial least squares discriminant analysis(OPLS-DA) enabled the identification of 21 metabolites as potential biomarkers that were relevant to phospholipid and energy metabolism. The results indicate that SM played a protective role against ISO-induced CVD in rats by regulating phospholipid and energy metabolic pathways. Further, we verified the apoptotic metabolic pathway using molecular biology methods, such as terminal deoxynucleotidyl transferase(TdT) deoxyuridine 5'-triphosphate(dUTP) nick-end labeling(TUNEL) assay and Western blot analysis. Furthermore, this study identified early biomarkers of CVD and elucidated the underlying mechanism of the therapeutic actions of SM, which is worth further to be investigated for development as a clinical therapy.     

α1A-亚型肾上腺素受体拮抗剂3D药效团模型的研究

运用Catalyst软件以34个α_1A-AR拮抗剂分子为训练集, 构建了包含一个氢键受体、一个正电中心和一个芳环中心的三元素药效团模型, 线性回归相关系数为0.89. 经13个分子组成的测试集验证该药效团模型具有较好的活性预测能力, 为寻找新的α_1A-AR拮抗剂分子提供了理论基础.     

Organotypic raft cultures as an effective in vitro tool for understanding Raman spectral analysis of tissue

There is a growing body of evidence showing that optical spectroscopy has the potential to be a useful in vivo diagnostic tool. Yet, so far there is no definitive cellular and biochemical understanding for the differences seen in the spectra from different tissue categories and disease states. In this study, we examine the use of organotypic raft cultures as an in vitro model of in vivo tissue conditions in an attempt to overcome some of the limitations of previously used methods. Organotypic raft cultures resembling normal and dysplastic epithelial cervical tissue were constructed and grown at an air-liquid interface for 2 weeks. Raman spectra of normal as well as dysplastic raft cultures were measured and compared with in vivo spectra from the corresponding tissue type. Histologic comparisons ensured that the raft cultures had similar structure and morphology to the corresponding intact tissue types. Raman spectra were also acquired from different layers of tissue. Spectral comparisons show that the Raman spectra of the raft cultures are similar to the spectra acquired from the cervix in vivo for both normal and dysplastic tissues. These results show that organotypic raft cultures are an effective and useful tool for the cellular and biochemical analysis of tissue spectroscopy.     

A Novel 3D Metal Organic Framework Based on an Azolate Ligand

Metal organic frameworks（MOFs） have attracted much attention over the world during the last several decades[1-3].Constructed from metal ions and organic building units,MOFs have great properties in gas adsorption[4],catalysis[5],luminescence[6] and magnetism[7].By varying the metal salts and organic building units,numerous MOF structures with designed functionalities and expected properties can be obtained.Among them,zeolitic imidazolate frameworks（ZIFs） have been investigated greatly due to their high thermal and chemical stabilities and tunable porosities[8].ZIFs can be obtained with imidazolate compounds as organic building units,such as imidazole,benzimidazole and 2-methylimidazole[9].Zeolite structures are composed of tetrahedral Si（Al）O4 units,with Si-O-Si angle ca.145°.ZIFs possess imidazole,with M-IM-M（M-metal,IM=imidazole） angle close to 145°,resembling zeolite（Scheme 1）[10].     

The Effect of Endothelial Cells on UVB‐induced DNA Damage and Transformation of Keratinocytes In 3D Polycaprolactone Scaffold Co‐culture System

Nitric oxide ( ${\mathbf{NO}}^{·}$) plays an important role in the regulation of redox balance in keratinocytes post‐UVB exposure. Since endothelial cells releases ${\mathbf{NO}}^{·}$ for a prolonged time post‐UVB, we determined whether human umbilical vein endothelial cells (HUVEC) could have an effect on UVB‐induced DNA damage and transformation of their adjacent keratinocytes (HaCaT) using a 3D cell co‐culturing system. Our data show that the levels of DNA breaks and/or cyclobutane pyrimidine dimer (CPD) along with γH2AX are higher in the co‐cultured than in the mono‐cultured keratinocytes post‐UVB. The ${\mathbf{NO}}^{·}$ level in the co‐cultured cells is increased approximately 3‐fold more than in mono‐cultured HaCaT cells within 1‐hour post‐UVB but then is reduced quickly in co‐cultured HaCaT cells comparing to mono‐cultured cells from 6 to 24 h post‐UVB. However, the peroxynitrite (ONOO^?) level is higher in the co‐cultured than in the mono‐cultured HaCaT cells in whole period post‐UVB. Furthermore, while expression level of inducible nitric oxide synthase (iNOS) is increased, the ratio of coupled/uncoupled eNOS is reduced in co‐cultured HaCaT cells compared to mono‐cultured HaCaT cells. Finally, the co‐cultured cells have a significantly increased transformation efficiency after repeating UVB exposure compared to mono‐culture HaCaT cells. Our results suggest that endothelial cells could enhance ${\mathbf{NO}}^{·}$/ONOO^? imbalance and promote transformation of adjacent keratinocytes.     

部分含氧、含氮有机分子的计算机三维结构模型

应用计算机软件CS Chem3D Pro3.5.2观察醇，酚，羧酸，胺和酰胺的典型化合物的三维模型和主要键参数，从氧，氮原子的杂化态入手，讨论它们的结构特征，进而应用于教学。     

Effect of Hydrolyzable Tannins on Glucose-Transporter Expression and Their Bioavailability in Pig Small-Intestinal 3D Cell Model

Intestinal transepithelial transport of glucose is mediated by glucose transporters, and affects postprandial blood-glucose levels. This study investigates the effect of wood extracts rich in hydrolyzable tannins (HTs) that originated from sweet chestnut (Castanea sativa Mill.) and oak (Quercus petraea) on the expression of glucose transporter genes and the uptake of glucose and HT constituents in a 3D porcine-small-intestine epithelial-cell model. The viability of epithelial cells CLAB and PSI exposed to different HTs was determined using alamarBlue^®. qPCR was used to analyze the gene expression of SGLT1, GLUT2, GLUT4, and POLR2A. Glucose uptake was confirmed by assay, and LC–MS/ MS was used for the analysis of HT bioavailability. HTs at 37 µg/mL were found to adversely affect cell viability and downregulate POLR2A expression. HT from wood extract Tanex at concentrations of 4 µg/mL upregulated the expression of GLUT2, as well as glucose uptake at 1 µg/mL. The time-dependent passage of gallic acid through enterocytes was influenced by all wood extracts compared to gallic acid itself as a control. These results suggest that HTs could modulate glucose uptake and gallic acid passage in the 3D cell model.     

Effects of Low‐Level Laser Therapy (LLLT) and Diclofenac (Topical and Intramuscular) as Single and Combined Therapy in Experimental Model of Controlled Muscle Strain in Rats

Muscle injuries represent ca 30% of sports injuries and excessive stretching of muscle causes more than 90% of injuries. Currently the most used treatments are nonsteroidal anti‐inflammatory drugs (NSAIDs), however, in last years, low‐level laser therapy (LLLT) is becoming an interesting therapeutic modality. The aim of this study was to evaluate the effect of single and combined therapies (LLLT, topical application of diclofenac and intramuscular diclofenac) on functional and biochemical aspects in an experimental model of controlled muscle strain in rats. Muscle strain was induced by overloading tibialis anterior muscle of rats. Injured groups received either no treatment, or a single treatment with topical or intramuscular diclofenac (TD and ID), or LLLT (3 J, 810 nm, 100 mW) 1 h after injury. Walking track analysis was the functional outcome and biochemical analyses included mRNA expression of COX‐1 and COX‐2 and blood levels of prostaglandin E₂ (PGE₂). All treatments significantly decreased COX‐1 and COX‐2 gene expression compared with injury group (P < 0.05). However, LLLT showed better effects than TD and ID regarding PGE₂ levels and walking track analysis (P < 0.05). We can conclude that LLLT has more efficacy than topical and intramuscular diclofenac in treatment of muscle strain injury in acute stage.     

Cancer Cell-targeted and Activatable Photoimmunotherapy Spares T Cells in a 3D Coculture Model

Photodynamic therapy (PDT) is an established therapeutic modality that uses nonionizing near-infrared light to activate photocytotoxicity of endogenous or exogenous photosensitizers (PSs). An ongoing avenue of cancer research involves leveraging PDT to stimulate antitumor immune responses; however, these effects appear to be best elicited in low-dose regimens that do not provide significant tumor reduction using conventional, nonspecific PSs. The loss of immune enhancement at higher PDT doses may arise in part from indiscriminate damage to local immune cell populations, including tumor-infiltrating T cells. We previously introduced “tumor-targeted, activatable photoimmunotherapy” (taPIT) using molecular-targeted and cell-activatable antibody–PS conjugates to realize precision tumor photodamage with microscale fidelity. Here, we investigate the immune cell sparing effect provided by taPIT in a 3D model of the tumor immune microenvironment. We report that high-dose taPIT spares 25% of the local immune cell population, five times more than the conventional PDT regimen, in a 3D coculture model incorporating epithelial ovarian cancer cells and T cells. These findings suggest that the enhanced selectivity of taPIT may be utilized to achieve local tumor reduction with sparing of intratumor effector immune cells that would otherwise be lost if treated with conventional PDT.     

Effects of Processing Parameters of 3D Bioprinting on the Cellular Activity of Bioinks

In this review, few established cell printing techniques along with their parameters that affect the cell viability during bioprinting are considered. 3D bioprinting is developed on the principle of additive manufacturing using biomaterial inks and bioinks. Different bioprinting methods impose few challenges on cell printing such as shear stress, mechanical impact, heat, laser radiation, etc., which eventually lead to cell death. These factors also cause alteration of cells phenotype, recoverable or irrecoverable damages to the cells. Such challenges are not addressed in detail in the literature and scientific reports. Hence, this review presents a detailed discussion of several cellular bioprinting methods and their process‐related impacts on cell viability, followed by probable mitigation techniques. Most of the printable bioinks encompass cells within hydrogel as scaffold material to avoid the direct exposure of the harsh printing environment on cells. However, the advantages of printing with scaffold‐free cellular aggregates over cell‐laden hydrogels have emerged very recently. Henceforth, optimal and favorable crosslinking mechanisms providing structural rigidity to the cell‐laden printed constructs with ideal cell differentiation and proliferation, are discussed for improved understanding of cell printing methods for the future of organ printing and transplantation.     

Ribes nigrum Leaf Extract Preferentially Inhibits IFN-γ-Mediated Inflammation in HaCaT Keratinocytes

Ribes nigrum L. (blackcurrant) leaf extracts, due to high levels of flavonols and anthocyanins, have been shown to exhibit beneficial effects in inflammatory diseases. However, whereas their traditional use has been investigated and validated in several models of inflammation and oxidative stress, the possible impact on skin disorders is still largely unknown. The purpose of this work was to elucidate the effects of R. nigrum leaf extract (RNLE) on keratinocyte-derived inflammatory mediators, elicited by a Th1 or Th2 cytokine milieu. HaCaT cells were challenged with TNF-α, either alone or in combination with the costimulatory cytokines IFN-γ or IL-4, and the release of proinflammatory cytokines and mediators (IL-8, IL-6, s-ICAM-1, and TSLP) was evaluated. The results showed that RNLE preferentially interferes with IFN-γ signaling, demonstrating only negligible activity on TNF-α or IL-4. This effect was attributed to flavonols, which might also account for the ability of RNLE to impair TNF-α/IL-4-induced TSLP release in a cAMP-independent manner. These results suggest that RNLE could have an antiallergic effect mediated in keratinocytes via mechanisms beyond histamine involvement. In conclusion, the discovery of RNLE preferential activity against IFN-γ-mediated inflammation suggests potential selectivity against Th1 type response and the possible use in Th1 inflammatory diseases.     

The Effects of UV Emission from Compact Fluorescent Light Exposure on Human Dermal Fibroblasts and Keratinocytes In Vitro

Compact fluorescent light (CFL) bulbs can provide the same amount of lumens as incandescent light bulbs, using one quarter of the energy. Recently, CFL exposure was found to exacerbate existing skin conditions; however, the effects of CFL exposure on healthy skin tissue have not been thoroughly investigated. In this study, we studied the effects of exposure to CFL illumination on healthy human skin tissue cells (fibroblasts and keratinocytes). Cells exposed to CFLs exhibited a decrease in the proliferation rate, a significant increase in the production of reactive oxygen species, and a decrease in their ability to contract collagen. Measurements of UV emissions from these bulbs found significant levels of UVC and UVA (mercury [Hg] emission lines), which appeared to originate from cracks in the phosphor coatings, present in all bulbs studied. The response of the cells to the CFLs was consistent with damage from UV radiation, which was further enhanced when low dosages of TiO₂ nanoparticles (NPs), normally used for UV absorption, were added prior to exposure. No effect on cells, with or without TiO₂ NPs, was observed when they were exposed to incandescent light of the same intensity.     

Boosting Charge Transport in a 2D/3D Perovskite Heterostructure by Selecting an Ordered 2D Perovskite as the Passivator

We demonstrate that an ordered 2D perovskite can significantly boost the photoelectric performance of 2D/3D perovskite heterostructures. Using selective fluorination of phenyl-ethyl ammonium (PEA) lead iodide to passivate 3D FA_0.8Cs_0.2PbI₃, we find that the 2D/3D perovskite heterostructures passivated by a higher ordered 2D perovskite have lower Urbach energy, yielding a remarkable increase in photoluminescence (PL) intensity, PL lifetime, charge-carrier mobilities (ϕμ), and carrier diffusion length (L_D) for a certain 2D perovskite content. High performance with an ultralong PL lifetime of ≈1.3 μs, high ϕμ of ≈18.56 cm² V⁻¹ s⁻¹, and long L_D of ≈7.85 μm is achieved in the 2D/3D films when passivated by 16.67 % para-fluoro-PEA₂PbI₄. This carrier diffusion length is comparable to that of some perovskite single crystals (>5 μm). These findings provide key missing information on how the organic cations of 2D perovskites influence the performance of 2D/3D perovskite heterostructures.     

D-Sorbitol Physical Properties Effects on Filaments Used by 3D Printing Process for Personalized Medicine

Fused filament fabrication (FFF) is a process used to manufacture oral forms adapted to the needs of patients. Polyethylene oxide (PEO) filaments were produced by hot melt extrusion (HME) to obtain a filament suitable for the production of amiodarone hydrochloride oral forms by FFF 3D printing. In order to produce personalized oral forms adapted to the patient characteristics, filaments used by FFF must be controlled in terms of mass homogeneity along filament. This work highlights the relation between filament mass homogeneity and its diameter. This is why the impact of filler excipients physical properties was studied. It has been showed that the particle’s size distribution of the filler can modify the filament diameter variability which has had an impact on the mass of oral forms produced by FFF. Through this work it was shown that D-Sorbitol from Carlo Erba allows to obtain a diameter variability of less than 2% due to its unique particle’s size distribution. Using the filament produced by HME and an innovating calibration method based on the filament length, it has been possible to carry out three dosages of 125 mg, 750 mg and 1000 mg by 3D printing with acceptable mass uniformity.