Discovery of new acetylcholinesterase inhibitors with small core structures through shape-based virtual screening.

Targeting acetylcholinesterase (AChE) using small molecule inhibitors is considered to be the most successful therapeutic strategy in the treatment of Alzheimer's disease (AD). Herein we present a shape-based virtual screening to identify new cores for the designing of AChE inhibitors. Ten active hits are identified and the most active hit, 5169-0032 and T5369186, showed comparable AChE inhibitory activity to tacrine. Prediction of physicochemical properties and ADME/T risk indicates their potential in druggability and safety. The two compounds provide new core and can serve as a promising fragment to design potent AChE inhibitors.

Expression of NEDD9 in pancreatic ductal adenocarcinoma and its clinical significance.

The aim of this study was to investigate the expression and prognostic significance of NEDD9 in pancreatic ductal adenocarcinoma (PDA). Expressional levels of NEDD9 mRNA and protein in paired pancreatic cancer lesions and adjacent noncancerous tissues were examined by quantitative real-time PCR and western blotting. NEDD9 expression was analyzed by immunohistochemistry in 106 patients with PDA. The correlations between NEDD9 immunostaining levels and clinicopathologic factors, as well as the follow-up data of patients, were analyzed statistically. NEDD9 protein and mRNA levels were elevated in pancreatic carcinoma lesions compared with the paired adjacent noncancerous tissues. A high level of expression of NEDD9 was significantly correlated with clinical staging (P < 0.001), lymph node metastasis (P < 0.001), and histological differentiation (P < 0.001). Patients with a higher NEDD9 expression had a significantly shorter survival time than those patients with lower NEDD9 expression. The multivariate analysis revealed that NEDD9 could serve as an independent factor of poor prognosis. Our finding indicates that NEDD9 could be used as prognostic molecular marker and therapeutic target for PDA.

Crucial Role of the Ca2+/CN Signaling Pathway in the Antifungal Activity of Seselin against Botrytis cinerea.

Botrytis cinerea causes gray mold in many fruit and vegetable crops. We previously found that Seselin (SL) displayed antifungal activity against B. cinerea (EC50 = 6.1 µg·mL-1), and this study investigated the effects of Ca2+ and the Ca2+/CN signaling pathway on its antifungal activity against B. cinerea. The results indicated that exogenous Ca2+, Cyclosporine A, and Verapamil reduced the sensitivity of SL against B. cinerea; SL significantly reduced the intracellular Ca2+ concentration in the hyphae; the sensitivity of strains ΔbcCCH1 and ΔbcMID1 to SL were significantly increased; and the expressions of CCH1, MID1, CNA, PMC1, and PMR1 genes of the Ca2+/CN signaling pathway were significantly downregulated by SL treatment. Hence, SL is a potential compound for developing fungicides against B. cinerea. SL dramatically reduces intracellular Ca2+ concentration and disturbs Ca2+ homeostasis, leading to cell death. The Ca2+/CN signaling pathway plays an important role in the antifungal activity of SL against B. cinerea.

Studies on chemical structure modification and structure-activity relationship of derivatives of gambogic acid at C(39).

The natural product gambogic acid exhibits high potency in inhibiting cancer cell lines. Rational medicinal modifications on gambogic acid will improve its physicochemical properties and drug-like characters. To investigate the structure-activity relationship of gambogic acid and also to find rational modification position on its chemical skeleton, we designed, synthesized, and characterized 16 derivatives of gambogic acid that were modified at C(39). The structure-activity relationships (SARs) were discussed. The anti-proliferation data were accquired through MTT (=3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assays of A549, BGC823, U251, HepG2, and MDA-MB-231 cancer cell lines. Most of the synthesized compounds showed strong inhibitory effects. The SAR study revealed that derivatives with aliphatic amino moieties at C(39) were more potent than those with other substituents. The C(39) position can undergo different kinds of chemical modifications without leading to loss of activity. Compounds 4 and 6 can serve as potential lead compounds for further development of new anticancer drugs.

Crucial role of Ca2+ /CN signalling pathway in the antifungal activity of disenecioyl-cis-khellactone against Botrytis cinerea.

BACKGROUND: Botrytis cinerea causes grey mould and is one of the most destructive fungal pathogens affecting important fruit and vegetable crops. In preliminary studies, we found that disenecioyl-cis-khellactone (DK) had strong antifungal activity against several fungi species including B. cinerea [half maximal effective concentration (EC50 ) = 11.0 µg mL-1 ]. In this study, we aimed to further evaluate the antifungal activity of DK against B. cinerea and determine the role of calcium ion/calcineurin (Ca2+ /CN) signalling pathway on its antifungal effect. RESULTS: DK was effective against B. cinerea in both in vitro and in vivo assays. Exogenous Ca2+ reduced the antifungal activity of DK. The combination of DK and cyclosporine A (CsA) did not exhibit an additive effect against B. cinerea. In contrast to CsA, DK reduced the intracellular Ca2+ concentration in B. cinerea. DK bound to calcineurin A (cnA) and up-regulated the expression of PMC1 and PMR1 genes. Moreover, DK sensitivity of △bccnA significantly decreased compared with that of Bc05.10 strain. CONCLUSION: DK is a promising lead compound for developing fungicides against B. cinerea. The Ca2+ /CN signalling pathway plays a crucial role in the DK antifungal activity, and cnA is one of the targets of DK against B. cinerea. DK directly reacts with cnA, which up-regulates the transcription of Ca2+ /CN-dependent target genes PMC1 and PMR1, decreasing the intracellular Ca2+ concentration and disturbing the intracellular Ca2+ balance, leading to cell death. © 2022 Society of Chemical Industry.

Anatomic measurement of wrist scaphoid and its clinical significance.

OBJECTIVE: To provide anatomical basis for the internal fixation of scaphoid fractures. METHODS: The shape and vascular lake of 48 dry scaphoids and 36 wet scaphoids were observed. RESULTS: The data of dry bone group and wet bone group were as follows: the height of scaphoid tubercle were (11.28+/-0.94) mm and (10.35+/-1.54) mm; the thickness of scapoid waist were (12.02+/-1.90) mm and (11.21+/-1.20) mm; the width of scapoid waist were (10.59+/-1.11) mm and (11.34+/-1.47) mm; the minimal thickness of the body of scapoid were (6.51+/-1.22) mm and (8.54+/-1.07) mm; the axis length of scapoid were (25.68+/-2.21) mm and (26.50+/-2.56) mm; the width of epicondyle of scaphoid of distal portion, waist and proximal portion were (6.50+/-1.06) mm, (5.14+/-1.01) mm, (4.42+/-1.16) mm and (6.64+/-1.18) mm, (6.01+/-0.75) mm and (5.71+/-0.78) mm, respestively. The main blood vessels came from the dorsal and the radial of wrist and passed through the whole scaphoid along the crest of scaphoid. CONCLUSION: The internal fixation of scaphoid can be designed according to the anatomical data without damaging the articular surface and blood supply.

The Effects of Different Catalysts, Substituted Aromatic Aldehydes on One-Pot Three-Component Biginelli Reaction.

AIM AND OBJECTIVE: The Biginelli reaction, first reported in 1893, is one great example of the important multicomponent reactions reported from 1893. Under the same conditions, the influence of the common catalysts on the yield of the Biginelli reaction was investigated. MATERIALS AND METHOD: To a round-bottom flask equipped with a spherical condenser were added 1,3- dicarbonyl compound (1.0 eq), urea (1.45 eq), aromatic aldehyde (1.0 eq), catalyst and methanol. The mixture was heated at reflux for 16 h. After cooling off, the mixture was filtered and washed with cold methanol to give DHPMs. Reaction solution was further purified by recrystallization with petroleum ether and ethyl acetate. Six catalytic systems, different 1,3-dicarbonyl compounds and different substituted aromatic aldehydes with varied substitutions are described for the Biginelli reaction. An analysis was also performed to study the factors that affect the yield of the reaction. RESULTS: When 1,3-dicarbonyl compound was ethyl acetoacetate, the CuCl/ conc.H2SO4 system gave the highest yield (90.5%). While when acetoacetamide was used, the yields of DHPMs in presence of PTSA/conc. HCl, conc. HCl or FeCl3•6H2O were all over 90%. Nine DHPMs with different substituents were obtained. CONCLUSION: The Lewis acid or mixed catalyst had no significant advantage over a single protonic acid as catalyst. Conc. HCl as the catalyst was found to be the most effective condition for the preparation of DHPMs. The aromatic aldehyde with weak electron-withdrawing substituent such as Br resulted in the best yield.

3'-Daidzein sulfonate sodium protects against memory impairment and hippocampal damage caused by chronic cerebral hypoperfusion.

3'-Daidzein sulfonate sodium (DSS) is a new synthetic water-soluble compound derived from daidzein, a soya isoflavone that plays regulatory roles in neurobiology. In this study, we hypothesized that the regulatory role of DSS in neurobiology exhibits therapeutic effects on hippocampal damage and memory impairment. To validate this hypothesis, we established rat models of chronic cerebral hypoperfusion (CCH) by the permanent occlusion of the common carotid arteries using the two-vessel occlusion method. Three weeks after modeling, rat models were intragastrically administered 0.1, 0.2, and 0.4 mg/kg DSS, once a day, for 5 successive weeks. The Morris water maze test was performed to investigate CCH-induced learning and memory deficits. TUNEL assay was used to analyze apoptosis in the hippocampal CA1, CA3 regions and dentate gyrus. Hematoxylin-eosin staining was performed to observe the morphology of neurons in the hippocampal CA1, CA3 regions and dentate gyrus. Western blot analysis was performed to investigate the phosphorylation of PKA, ERK1/2 and CREB in the hippocampal PKA/ERK1/2/CREB signaling pathway. Results showed that DSS treatment greatly improved the learning and memory deficits of rats with CCH, reduced apoptosis of neurons in the hippocampal CA1, CA3 regions and dentate gyrus, and increased the phosphorylation of PKA, ERK1/2, and CREB in the hippocampus. These findings suggest that DSS protects against CCH-induced memory impairment and hippocampal damage possibly through activating the PKA/ERK1/2/CREB signaling pathway.

Evaluation of antifungal activities and structure-activity relationships of coumarin derivatives.

BACKGROUND: Osthol is a natural coumarin and lead compound that has been developed into commercial fungicides in China. Natural coumarins comprise five major subtypes: simple coumarins, linear furanocoumarins, angular furanocoumarins, linear pyranocoumarins and angular pyranocoumarins. Studies pertaining to the antifungal activities of linear pyranocoumarins are few, and no reports exist for the antifungal activities of angular pyranocoumarins. In order to discover more antifungal natural coumarins, we synthesised a series of simple natural coumarins and isolated several plant-based furanocoumarins and pyranocoumarins using previously described methods. The compounds were biologically evaluated against some plant fungal pathogens. RESULTS: Several of the 35 coumarins evaluated here exhibited strong activities against specific fungal species, including compound 25 (Pd-D-V, a linear pyranocoumarin), compound 26 (libanorin, an angular furanocoumarin) and compound 34 (disenecioyl khellactone, an angular pyranocoumarin). Compound 25 exhibited a high activity against Sclerotinia sclerotiorum (EC50 = 13.2 µg mL-1 ); compound 34 displayed a strong antifungal activity against Botrytis cinerea (EC50 = 11.0 µg mL-1 ). CONCLUSION: This study demonstrates that several natural coumarins (one linear pyranocoumarin and one angular pyranocoumarin in particular) exhibit strong antifungal activities. These results call for further studies, where these coumarins can be examined as potential lead compounds for developing novel antifungal agents. © 2016 Society of Chemical Industry.

TLR4 polymorphisms associated with developing gastric pre-cancer lesions in a Chinese Han population.

Helicobacter pylori infection is a risk factor for gastric cancer. In addition, toll-like receptor 4 (TLR4) plays a fundamental role in pathogen recognition and activation of innate immunity. This study investigated the association of TLR4 polymorphisms with a risk of intestinal metaplasia (IM) and intraepithelial neoplasia (IN) in a Chinese Han population. This study analyzed TLR4 gene polymorphisms in 333 patients (IM, 193 cases; IN, 140 cases) and 312 atypia-free controls in a Chinese Han population using a Taqman allelic discrimination assay. The TLR4 single nucleotide polymorphisms +896A/G and +1196C/T were not associated with the risk of IM or IN. However, the single-locus analysis showed that the C allele of TLR4+2856T/C had significantly reduced risk of IM and IN [adjusted odds ratio (OR)=0.42; 95%CI=0.29-0.62 and OR=0.62; 95%CI=0.41-0.93, respectively] compared with the wild-type homozygote (TT). The frequencies of TLR4+2856T/C TC and T carrier were significantly lower in patients with Sydney's slight IM and low grade IN (P<0.01 and P=0.01, respectively), while the TC genotype showed a lower risk of moderate IM compared to healthy controls (P=0.045). In addition, the data revealed that H. pylori infection, heavy alcohol consumption and high salt uptake were associated with a higher susceptibility for developing this neoplasm. TLR4 rs10759932 TC and C carriers were associated with a lower risk in developing precancerous lesions in the stomach in a Chinese Han population.

Vagal afferents mediate antinociception of estrogen in a rat model of visceral pain: the involvement of intestinal mucosal mast cells and 5-hydroxytryptamine 3 signaling.

UNLABELLED: Estrogen reportedly facilitates visceral nociception at the spinal or supraspinal level. The present study was aimed to investigate whether estrogen modulates visceral pain through the vagal pathway. Ovariectomized rats received estradiol, which was administered subcutaneously (to act through both the vagal and spinal pathways) or intraduodenally (to preferentially act through the vagal pathway). Luminally applied estradiol induced a rapid and significant decrease in the visceromotor response to colorectal distension, with increased c-Fos expression in nodose ganglion neurons. Systemically injected estradiol increased visceromotor response and c-Fos expression in both nodose and dorsal root ganglion (T6-12) neurons. The antinociceptive effect of estrogen was abolished by surgical vagotomy or chemical denervation of vagal afferents. Both luminally and systemically administered estradiol elicited selective 5-hydroxytryptamine secretion from the duodenum. Granisetron, a 5-hydroxytryptamine 3 receptor antagonist, reversed the antinociceptive effect of estrogen. Intestinal mucosal mast cell stabilizers prevented estradiol-induced antinociception and 5-hydroxytryptamine secretion. Ultrastructural analysis revealed that estradiol caused piecemeal degranulation of intestinal mucosal mast cells. The actions of estradiol were inhibited by an estrogen receptor ß antagonist and mimicked by an estrogen receptor ß agonist. These results suggest that estrogen can trigger vagus-mediated antinociception, which is masked by its spinally mediated pronociception. PERSPECTIVE: This study is the first to show a vagus-mediated estrogenic antinociception, in which the nongenomic estrogen receptor ß-mediated, intestinal mucosal mast cell-derived 5-hydroxytryptamine/5-hydroxytryptamine 3 receptor pathway is involved. This work may provide new insights into the sex hormone modulation of visceral sensitivity related to irritable bowel syndrome and indicate potential therapeutic targets to manage this disease.

[Anatomical study of improved lateral arm free flap].

OBJECTIVE: To study the anatomy and preparation methods of an improved lateral arm free flap (LAFF) for the future clinical application. METHODS: Twenty-two adult upper extremities from cadavers after injected with red latex through common carotid arteries were used. The course, branches, distribution and variations of the blood vessels and nerves of the improved LAFF were observed. The outer diameters of the vessels were measured. RESULTS: The mean length of vascular pedicle of the improved LAFF was (14.85 ± 1.28) cm, significantly more than that (5.46 ± 2.60) of traditional LAFF (t = -8.483, P < 0.001). The mean outer diameters of pedicle arteries and veins in the improved LAFF were (2.24 ± 0.66) mm and (2.22 ± 0.52) mm, significantly more than those (1.15 ± 0.21 and 1.26 ± 0.23) in traditional LAFF (t = -8.690, P < 0.001; t = -15.057, P < 0.001), respectively. CONCLUSION: The improved LAFF has a longer vascular pedicle and larger artery and vein in diameter than conventional LAFF, and is more suitable for the repair of the small and medium-sized defects of the head and neck.

[The design of inferior rectus abdominis flap and its study of applied anatomy].

OBJECTIVE: To describe the anatomy of inferior epigastric artery and its correlation with rectus abdominis muscle in order to provide anatomical basis for reconstruction of defect in the oral and maxillofacial region. METHODS: 10 Chinese adult specimens fixed by 10% formalin were simulated for the design of inferior rectus abdominis flap, and then observing, taking a photograph during the cadavers were being operated and measuring by image analysis software in the end. RESULTS: The original external diameters of inferior epigastric artery and length of its trunk were (2.58 +/- 0.60) mm and (85.87 +/- 14.90) mm respectively. Its perforators distribute intensively toward hilum. CONCLUSION: Whether rectus abdominis myocutaneous flap or deep inferior epigastric perforator flap adapts to reconstruct the defect of oral and maxillofacial region due to invariable traveling of inferior epigastric artery and its wide original external diameters relatively.

[Applied anatomy study on the reversed submental island flap].

OBJECTIVE: To provide anatomic data for clinical use of reversed submental island flap in reconstruction of buccal and facial defects. METHODS: Twenty cadavers with 40 sides of head and neck which were fixed by formalin and perfused by red emulsion, 6 fresh cadavers which were perfused by coloring agent were dissected, observed and measured. RESULTS: The diameter of the submental artery, the superior lip artery and the inferior lip artery at their origins were respectively (1.42 +/- 0.30) mm (x +/- s, same at below), (1.34 +/- 0.35) mm and (1.34 +/- 0.27) mm respectively. The realistic distance from the origin of inferior and superior lip arteries to the origin of submental artery were (50.13 +/- 13.79) mm and (92.09 8.73) mm, respectively. However, the straight distances from the origin of inferior and superior lip arteries to the origin of submental artery were (35.19 +/- 9.18) mm, (64.99 +/- 5.24) mm, respectively. There were rich anastomoses between both facial arteries, and the facial arteries also anastomosed richly with the ophthalmic artery and the superficial temporal artery. On the marginal mandibular plane, the facial vein ran closely with the artery, the marginal mandibular branch of the facial nerve crossed superficially to the facial blood vessels; superior to this plane, the vein ran 1 cm behind the artery. The buccal branches of the facial nerve crossed superficially to the facial vein and ran into the muscles behind or across the facial artery. The facial vein anastomosed richly with the ophthalmic vein and the maxillary vein. CONCLUSION: There is anatomic feasibility for the reconstruction of buccal and facial defects by using pedicled submental reversed island flap. The reversing point of the flap ought to be at the cheilion plane or inferiorly. Anatomy, regional