The G protein Gαs acts as a tumor suppressor in sonic hedgehog signaling-driven tumorigenesis

G protein-coupled receptors (GPCRs) are critical players in tumor growth and progression. The redundant roles of GPCRs in tumor development confound effective treatment; therefore, targeting a single common signaling component downstream of these receptors may be efficacious. GPCRs transmit signals through heterotrimeric G proteins composed of Gα and Gβγ subunits. Hyperactive Gα_s signaling can mediate tumor progression in some tissues; however, recent work in medulloblastoma and basal cell carcinoma revealed that Gα_s can also function as a tumor suppressor in neoplasms derived from ectoderm cells including neural and epidermal stem/progenitor cells. In these stem-cell compartments, signaling through Gα_s suppresses self-renewal by inhibiting the Sonic Hedgehog (SHH) and Hippo pathways. The loss of GNAS, which encodes Gα_s, leads to activation of these pathways, over-proliferation of progenitor cells, and tumor formation. Gα_s activates the cAMP-dependent protein kinase A (PKA) signaling pathway and inhibits activation of SHH effectors Smoothened-Gli. In addition, Gα_s-cAMP-PKA activation negatively regulates the Hippo pathway by blocking the NF2-LATS1/2-Yap signaling. In this review, we will address the novel function of the signaling network regulated by Gα_s in suppression of SHH-driven tumorigenesis and the therapeutic approaches that can be envisioned to harness this pathway to inhibit tumor growth and progression.     

Nondestructive evaluation of photosynthesis by delayed luminescence in Arabidopsis in Petri dishes

Nondestructive evaluation of photosynthesis is a valuable tool in the field and laboratory. Delayed luminescence (DL) can reflect charge recombination through the backflow of electrons. However, DL detection has not yet been adapted for whole plants in Petri dishes. To compensate for differences in DL decay between sibling Arabidopsis plants grown under the same conditions, we developed a time-sequential double measurement method. Using this method, we examined the influence of photosynthetic electron flow inhibitors, and differences in the DL decay curves were categorized by considering the initial and late phases of the decay curves, as well as their intermediate slopes. The appearance of concavity and convexity in DL curves in Arabidopsis was different from unicellular algae, suggesting complexity in the photosynthetic machinery of higher plants. This detection method should be invaluable for evaluating photosynthetic defects in higher plants under sterile conditions without interrupting plant culture.     

Synthesis and Th1-immunostimulatory activity of α-galactosylceramide analogues bearing a halogen-containing or selenium-containing acyl chain

A novel series of CD1d ligand α-galactosylceramides (α-GalCers) were synthesized by incorporation of the heavy atoms Br and Se in the acyl chain backbone of α-galactosyl-N-cerotoylphytosphingosine. The synthetic analogues are potent CD1d ligands and stimulate mouse invariant natural killer T (iNKT) cells to selectively enhance Th1 cytokine production. These synthetic analogues would be efficient X-ray crystallographic probes to disclose precise atomic positions of alkyl carbons and lipid–protein interactions in KRN7000/CD1d complexes.     

Molecular docking studies and synthesis of novel bisbenzimidazole derivatives as inhibitors of α-glucosidase

A series of bisbenzimidazole derivatives starting from o-phenylenediamine and 4-nitro-o-phenylenediamine were prepared with oxalic acid. Most of the reactions were conducted using both the microwave and conventional methods to compare yields and reaction times. The operational simplicity, environmental friendly conditions and high yield in a significantly short reaction time were the major benefits. All substances’ inhibitory activities against α-glucosidase were evaluated. The results may suggest a significant role for the nature of bisbenzimidazole compounds in their inhibitory action against α-glucosidase. They showed different range of α-glucosidase inhibitory potential with IC₅₀ value ranging between 0.44 ± 0.04 and 6.69 ± 0.01 μM when compared to the standard acarbose (IC₅₀, 13.34 ± 1.26 μM). This has described a new class of α-glucosidase inhibitors. Molecular docking studies were done for all compounds to identify important binding modes responsible for inhibition activity of α-glucosidase.     

小花草玉梅正常和自然变异植株的AP3-3基因研究

野生小花草玉梅(Anemone rivularis var.flore-minore)正常植株和花被片自然变异植株的外观形态差异很大,该研究以二者为材料,利用常规PCR和高效热不对称PCR(Hi-Tail PCR)技术从其正常和变异植株的基因组中各分离得到1个B类基因。序列分析证明,二者隶属于B类MADS-box基因AP3家族的旁系同源基因AP3-3分枝,分别命名为NArAP3-3(正常植株)和VArAP3-3(变异植株)。NArAP3-3基因全长3 795bp,VArAP3-3基因全长3 898bp,二者均含有1个666bp的开放阅读框(ORF),可编码221个氨基酸,具有典型的植物MADS-box基因结构,其编码肽链包含了MADS区、K区、Ⅰ区和C区。对比NArAP3-3和VArAP3-3基因的全长序列,发现VArAP3-3基因比NArAP3-3多了1段49bp的插入,且在ORF序列与NArAP3-3基因相比有4个碱基突变。对二者的全长序列、所编码的221个氨基酸及插入序列的生物信息学分析显示,二者在基因启动子、蛋白质基本性质、结构功能域、二级三级预测结构等方面均有差异,推测这些差异可能是花被片变异产生的原因之一。该研究结果为进一步探索其变异机制奠定了基础。     

Drugging the undruggable: gabapentin,pregabalin and the calcium channel α2δ subunit

In the post-genomic era, the idea of using the sequence of a protein to determine its potential role as a drug target has gained currency. The goal of this approach to drug discovery is to use the sequence of a protein that is known to bind a specific ligand or drug, along with the known structure of the ligand binding site, to predict other similar proteins that are also “druggable”. Gabapentin (Neurontin) and pregabalin (Lyrica) are drugs currently in the clinic that were developed based on the hypothesis that generating non-hydrolyzable analogs of GABA would lead to the development of antiepileptic agents. While these compounds are indeed good anticonvulsants, their activity is surprisingly not due to activity in the GABAergic system. By purifying the protein to which gabapentin bound, and determining its identity as the α₂δ₁ subunit of voltage gated calcium channels, it was possible to make progress in developing new compounds with similar activities to gabapentin, including pregabalin. The recognition of the α₂δ₁ subunit as the receptor for these drugs also meant that related proteins, such as α₂δ₃, may be interesting targets for novel pain therapeutics.     

CT灌注成像在兔早期肝硬化诊断中的应用价值研究

目的:探讨CT灌注成像在兔早期肝硬化诊断中的应用价值。方法:将55只新西兰大白兔随机分为2组,其中实验组45只,对照组10只。实验组给予皮下注射葡萄籽油稀释的50%CCL4,1次/4天,前4次剂量为1.0 m L/kg,第5次剂量为1.35 m L/kg,共注射20次。对照组采用同样方法只注射相同剂量的生理盐水。每注射4次后分别对实验组兔7只和正常对照组兔2只做螺旋CT灌注扫描,分析灌注参数,同时做相应的病理学观察,将二者进行比较及统计学分析。结果:注药4次末,兔血清ALT及AST明显高于注药前,注药8次末,兔血清ALT及AST最高,之后兔血清ALT及AST轻度减低,注药前后兔血清ALT及AST的变化有统计学意义(P0.05)。而血清(ALB)水平变化不明显,仅在注药16次末后,ALB水平稍减低,但差异无统计学意义。对照组肝脏灌注参数正常,实验组从注药4次开始,HAP呈上升趋势,但注药4次末及注药8次末,实验组及对照组之间差异无统计学意义(P0.1),注药12次末后二者之间差异有统计学意义(P0.05);而HPP、HBF及HBV呈下降趋势,MTT逐渐延长,与对照组的差异均有统计学意义(P0.05)。随兔血清ALT及AST的升高,HAP逐渐升高,MTT逐渐延长,而HPP、HBF及HBV逐渐减低。实验组肝小叶正常结构破坏,肝实质被纤维组织分割成大小不一、圆形或近圆形结节(假小叶),间隔较窄,炎症轻,结节边界尚整齐;汇管区内门脉小支扩张,壁增厚。对照组肝小叶结构规整,肝板排列有序,汇管区无扩大,其内个别炎细胞浸润,肝小叶内偶见点灶状坏死。结论:全肝CT灌注功能成像可为早期肝硬化的诊断提供影像学依据,将灌注征象与病理学变化结合有利于肝硬化的早期诊断和治疗。     

胃癌根治术后腹腔热灌注化疗对患者血清CEA、CA19-9水平及免疫功能的影响

目的:探讨胃癌根治术后腹腔灌注化疗对患者血清CEA(血清癌胚抗原,carcinoembryonic antigen)、CA19-9(糖链抗原19-9,carbohydrate antigen19-9)水平及免疫功能的影响。方法:回顾性2015年2月至2017年4月我院收治的胃癌患者临床资料,依据接受治疗方案不同分为全身静脉化疗组(对照组)和全身静脉化疗联合腹腔热灌注化疗组(观察组),每组各41例。检测和比较两组患者化疗前(治疗前)与化疗1个月后(治疗后)血清肿瘤标志物CEA、CA19-9与免疫功能指标水平的变化,治疗后毒副作用发生情况及治疗前后生活质量的改善情况。结果:治疗前,两组间血清CEA、CA19-9、CD3~+、CD4~+、CD8~+、CD4+/CD8~+水平比较差异均无统计学意义(P0.05);观察组治疗后血清CEA、CA19-9及CD8~+水平显著低于对照组,CD3~+、CD4~+、CD4~+/CD8~+水平显著高于对照组,差异有统计学意义(P0.05);两组骨髓抑制、恶心呕吐、腹痛腹泻及肠梗发生率比较差异均不显著无统计学意义(P=0.478,0.668,0.315,0.552);观察组生活质量改善总有效率为85.37%,显著高于对照组(70.73%,P=0.017)。结论:与单纯全身化疗相比,胃癌根治术后腹腔灌注化疗可更有效降低患者血清CEA、CA19-9水平,改善患者免疫功能,提高其生活质量,且安全性较高。     

INH-α对BMP9诱导间充质干细胞成骨分化的作用

为了研究抑制素α亚基(inhibinα-subunit INH-α)对骨形态发生蛋白9(bone morphogenetic protein9,BMP9)诱导的间充质干细胞(mesenchymal stem cells,MSCs)成骨分化的影响,本研究采用细胞化学染色法检测第3天、第5天、第7天细胞中碱性磷酸酶(alkaline phosphatase,ALP)活性的变化。利用RT-PCR和Western blotting检测细胞中的成骨分化早期标志物(Runx2)和晚期标志物(OPN)的mRNA含量及蛋白表达水平。茜素红S染色法检测第21天细胞中的钙盐沉积变化。发现BMP9组ALP活性明显增高,INH-α组ALP活性与对照组相比无明显变化,但联合运用BMP9和INH-α组ALP活性较BMP9组明显降低。此外,BMP9组Runx2和OPN的mRNA含量和蛋白表达水平明显增高,而联用BMP9和INH-α组中的Runx2和OPN水平较BMP9组显著下降(p<0.01)。同样,在茜素红S染色实验中,BMP9组钙盐结节明显增多,染色深;而在联合运用BMP9和INH-α组钙盐结节较BMP9组明显减少,染色变浅。说明INH-α能够抑制BMP9诱导间充质干细胞成骨分化作用。