Search

Objective: To compare the clinical outcomes of staged total knee arthroplasty (TKA) performed on both knees in the same patient using gap balancing (GB) and measured resection (MR) techniques, respectively. Methods: The clinical data of 57 patients undergoing bilateral staged TKA at the Xi'an Jiaotong University Affiliated Honghui Hospital from July 2018 to January 2020 were analyzed. Using the random number table, MR or GB technique was selected when patients underwent primary TKA, and contralateral procedure was done with another technique. The procedures were performed by one chief surgeon, and the same prosthesis was chosen for all the procedures. The two osteotomy techniques for TKA were compared in terms of surgical status, radiographic data, functional recovery and satisfaction rate. Results: Total of 57 patients, including 16 males and 41 females, were included in the study with a mean age of (68.5±4.6) years (59-79 years) at primary TKA. All patients were followed up for (29.6±4.5) months (22-39 months). The interval between the two procedures was (4.7±3.0) months (0.5-12.0 months). Postoperative drainage was less in the GB side when compared with that in the MR side [(93.6±22.2) ml vs (109.9±36.9) ml, P =0.003]. At the 1-month postoperative follow-up, the visual analogue scale (VAS) of pain was lower on the GB side (3.0±0.8) than on the MR side (3.5±1.2), the range of motion (ROM) was higher on the GB side (105.7°±8.2° vs 100.2°±7.5°), the Knee Society Score (KSS) was higher on the GB side (78.5±5.4 vs 74.2±6.3), and the Western Ontario and McMaster University (WOMAC) score was lower on the GB side (35.4±5.5 vs 38.0±6.3), there were significant differences in the up-mentioned indexes between the two groups (all P <0.05). However, the repeated-measures analysis of variance indicated that there was no significant difference in VAS score, ROM, KSS score and WOMAC score between the two techniques (all P >0.05). The satisfactory rate of GB technique was 84.2%(48/57), ant it was 86.0%(49/57) with MR technique ( P =0.446). There was also no significant difference between the two techniques in terms of complications ( P =0.754). Conclusion: Both the GB and MR technique result in good knee function with similar clinical outcomes in patients receiving TKA in both knees for osteoarthritis without significant deformity.

Background: The frequency of acute hypertriglyceridemic pancreatitis (AHTGP) is increasing worldwide. AHTGP may be associated with a more severe clinical course and greater mortality than pancreatitis caused by other causes. Early identification of patients with severe inclination is essential for clinical decision-making and improving prognosis. Therefore, we first developed and validated a risk prediction score for the severity of AHTGP in Chinese patients., Aim: To develop and validate a risk prediction score for the severity of AHTGP in Chinese patients., Methods: We performed a retrospective study including 243 patients with AHTGP. Patients were randomly divided into a development cohort ( n = 170) and a validation cohort ( n = 73). Least absolute shrinkage and selection operator and logistic regression were used to screen 42 potential predictive variables to construct a risk score for the severity of AHTGP. We evaluated the performance of the nomogram and compared it with existing scoring systems. Last, we used the best cutoff value (88.16) for severe acute pancreatitis (SAP) to determine the risk stratification classification., Results: Age, the reduction in apolipoprotein A1 and the presence of pleural effusion were independent risk factors for SAP and were used to construct the nomogram (risk prediction score referred to as AAP). The concordance index of the nomogram in the development and validation groups was 0.930 and 0.928, respectively. Calibration plots demonstrate excellent agreement between the predicted and actual probabilities in SAP patients. The area under the curve of the nomogram (0.929) was better than those of the Bedside Index of Severity in AP (BISAP), Ranson, Acute Physiology and Chronic Health Evaluation (APACHE II), modified computed tomography severity index (MCTSI), and early achievable severity index scores (0.852, 0.825, 0.807, 0.831 and 0.807, respectively). In comparison with these scores, the integrated discrimination improvement and decision curve analysis showed improved accuracy in predicting SAP and better net benefits for clinical decisions. Receiver operating characteristic curve analysis was used to determine risk stratification classification for AHTGP by dividing patients into high-risk and low-risk groups according to the best cutoff value (88.16). The high-risk group (> 88.16) was closely related to the appearance of local and systemic complications, Ranson score ≥ 3, BISAP score ≥ 3, MCTSI score ≥ 4, APACHE II score ≥ 8, C-reactive protein level ≥ 190, and length of hospital stay., Conclusion: The nomogram could help identify AHTGP patients who are likely to develop SAP at an early stage, which is of great value in guiding clinical decisions., Competing Interests: Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Gomisin A is a dietary lignan compound isolated from the fruit of Schisandra chinensis and has many pharmacological properties, including hepato-protective, anti-diabetic, and anti-oxidative activities. However, the benefit of gomisin A is still not well understood. The action of gomisin A is diverse. However, the effect of gomisin A on Ca 2+ signaling in prostate cancer cells is unknown. Ca 2+ is a pivotal second envoy that triggers and regulates cellular processes such as apoptosis, fertilization, energy transduction, secretion, and protein activation. The goal of this study was to explore the action of gomisin A on [Ca 2+ ] i and cytotoxicity in PC3 prostate cancer cells. Gomisin A at 100-200 μM provoked [Ca 2+ ] i raises. 20% of the response was reduced by removing external Ca 2+ . The Ca 2+ influx provoked by gomisin A was suppressed by 20% by store-caused Ca 2+ entry suppressors: econazole, SKF96365, nifedipine; also by phorbol 12-myristate 13 acetate and GF109203X. Without external Ca 2+ , gomisin A-caused [Ca 2+ ] i raises were abolished by thapsigargin. In contrast, gomisin A suppressed the [Ca 2+ ] i raises caused by thapsigargin. U73122 fell short to change gomisin A-caused [Ca 2+ ] i responses. Gomisin A (20-100 μM) elicited cytotoxicity in a dose-associated fashion. Blockade of [Ca 2+ ] elevations with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl failed to inhibit cytotoxicity of gomisin A. Collectively, gomisin A evoked [Ca 2+ ] i raises and provoked cytotoxicity in a Ca 2+ -dissociated fashion in prostate cancer cells., Competing Interests: None

Wang-Bi capsule (WB) is a traditional Chinese medicine (TCM)-based herbal formula, and it has been used in the treatment of rheumatoid arthritis (RA) in China for many years. Additionally, WB is also used as a supplement to the treatment of osteoarthritis (OA) in clinical practice. Our research aimed to reveal the therapeutic effects and underling mechanism of WB on RA and OA through computational system pharmacology analysis and experimental study. Based on network pharmacology analysis, a total of 173 bioactive compounds interacted with 417 common gene targets related to WB, RA, and OA, which mainly involved the PI3K-Akt signaling pathway. In addition, the serine-threonine protein kinase 1 (AKT1) might be a core gene protein for the action of WB, which was further emphasized by molecular docking. Moreover, the anti-inflammatory activity of WB in vitro was confirmed by reducing NO production in lipopolysaccharide (LPS)-induced RAW264.7 cells. The anti-RA and OA effects of WB in vivo were confirmed by ameliorating the disease symptoms of collagen II-induced RA (CIA) and monosodium iodoacetate-induced OA (MIA) in rats, respectively. Furthermore, the role of the PI3K-Akt pathway in the action of WB was preliminarily verified by western blot analysis. In conclusion, our study elucidated that WB is a potentially effective strategy for the treatment of RA and OA, which might be achieved by regulating the PI3K-Akt pathway. It provides us with systematic insights into the effects and mechanism of WB on RA and OA., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

Hepatotoma is the leading type of primary liver cancer in adults and third cause of death in the world. Hydroxytyrosol is a natural phenol existing in olive (Olea europaea L.). Hydroxytyrosol is the chief ingredient of olive oil, which was early deemed to be the most robust antioxidant in olive oil. Hydroxytyrosol is known to inhibit various types of cancer by different methods. This study was aimed to delineate the action of hydroxytyrosol on viability and [Ca 2+ ] i in HepG2 hepatoma cells. Fura-2 was used to detect [Ca 2+ ] i , and WST-1 assays were applied to explore cell cytotoxicity. Hydroxytyrosol elicited [Ca 2+ ] i raises. Eliminating external Ca 2+ diminished the Ca 2+ signal by 30%. Hydroxytyrosol-evoked Ca 2+ influx was diminished by 20% by three inhibitors of store-operated Ca 2+ channels and by a protein kinase C activator and an inhibitor. In the absence of Ca 2+ , thapsigargin eradicated hydroxytyrosol-provoked [Ca 2+ ] i raises. Suppression of phospholipase C (PLC) with U73122, a PLC inhibitor, did not inhibit hydroxytyrosol-elicited [Ca 2+ ] i raises. Hydroxytyrosol reduced cell viability. This cytotoxic action was not reversed by preincubation with BAPTA/AM, a cytosolic Ca 2+ binder. In sum, in HepG2 hepatoma cells, hydroxytyrosol elicited [Ca 2+ ] i raises by provoking PLC-unrelated discharge of Ca 2+ from ER and Ca 2+ influx through PKC-sensitive store-operated Ca 2+ entry. In addition, hydroxytyrosol elicited Ca 2+ -dissociated cytotoxicity., Competing Interests: None

Objective: Adding vitamin E to highly cross-linked polyethylene liners is frequently performed in clinical practice, aiming at reducing liner wear, increasing liner survival, and delaying revision surgery. This study is aimed at evaluating the revision rate, total femoral head penetration, and postoperative clinical function of highly cross-linked polyethylene liners with and without vitamin E in total hip arthroplasty., Methods: We conducted a systematic literature search to identify the use of highly cross-linked vitamin E liners compared to other liners in patients who received total hip arthroplasty (THA) before April 2021. The study quality assessment and data collection were conducted by two independent reviewers. Studies were artificially grouped, and vitamin E-enhanced liners (VE-PE) were compared with vitamin E-free liners (non-VE-PE). Analyses were executed using Review Manager version 5.4.1., Results: From the preliminary screening of 568 studies, fourteen studies met the research criteria. Compared to non-VE-PE, using VE-PE reduced the all-cause revision rate (odds ratio = 0.54; 95% confidence interval (CI) 0.40, 0.73; P < 0.0001). The total femoral head penetration of the VE-PE was lower than that of the non-VE-PE (mean difference = -0.10; 95% CI -0.17, -0.03; P = 0.007). However, there was no difference in clinical function, including the Harris Hip Score and EuroQol Five-Dimension Questionnaire scores., Conclusion: Compared to the liners without vitamin E, the addition of vitamin E to liners could reduce the all-cause revision rate by approximately 46% in the short-term follow-up. In addition, even though addition of vitamin E could also slow down femoral head penetration, there is no contribution to clinical function., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Qian-Yue Cheng et al.)

Thin-walled cylindrical shell storage tanks are pressure vessels in which the walls of the vessel have a thickness that is much smaller than the overall size of the vessel. These types of structures have global applications in various industries, including oil refineries and petrochemical plants. However, these storage tanks are vulnerable to fire and explosions. Therefore, a parametric study using numerical simulation was carried out, considering the internal liquid level, wall thickness, material yield strength, constraint conditions, and blast intensity, with a diameter of 100 m and height of 22.5 m under different blast loads using the finite element analysis method. The thickness of the tank wall is varied as 10 mm, 20 mm, 30 mm, and 40 mm, while the fill level of internal fluid is varied as 25, 50, 75, and 100%. The blast simulation was conducted using LS-DYNA software. The numerical results are then compared with analytical results. The effects of blast intensity, standoff distance, wall thickness, and fill level of internal fluid on the structural behaviour of the storage tank were investigated and discussed.

Objective: This study was designed to investigate preoperative anxiety situations and postoperative pain degree in Chinese patients undergoing laparoscopic hysterectomy and to analyze the related factors of preoperative anxiety and the correlation between preoperative anxiety and postoperative pain to provide a reference for effective postoperative analgesia management. Methods: A total of 100 female patients undergoing laparoscopic hysterectomy were enrolled in this study and randomly divided into two groups ( n = 50, each). In group A, the patients were treated with dexmedetomidine and sufentanil for postoperative analgesia. In group B, the patients were treated with sufentanil alone for postoperative analgesia. All patients were evaluated with a self-rating anxiety scale (SAS) 1 day before the operation. The patients' pain was evaluated using the numerical rating scale (NRS) 1 day after the operation, and data were recorded. Results: In these 100 patients, the highest preoperative SAS score was 48, and the average score was 40.99 ± 4.55 points, which is higher than the norm in China. There were significant differences in preoperative SAS scores among patients with different occupations and previous surgical experience ( P < 0.05). There was no significant difference in SAS scores among patients with different education levels ( P > 0.05). The postoperative NRS score of group A was significantly higher than that of group B, and the difference was statistically significant ( P < 0.05). The correlation coefficients between SAS scores and NRS scores in groups A and B were 0.836 and 0.870, respectively, presenting with a significantly positive correlation. Conclusion: Preoperative anxiety is an important predictor of postoperative pain. Patients undergoing laparoscopic hysterectomy have preoperative anxiety. The degree of anxiety is influenced by the occupation and previous operation experience of the patients, and patients with higher preoperative anxiety have greater postoperative pain. In addition, we should not neglect the management of postoperative pain because of the small trauma of laparoscopic surgery, and dexmedetomidine combined with sufentanil can improve the postoperative analgesic effect., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhang, Hao, Hou, Wu, Jing and Sun.)

Cornual heterotopic pregnancy is an extremely rare, life-threatening complication during pregnancy. Here, we report a 33-year-old woman who suffered cornual heterotopic pregnancy after in vitro fertilization embryo transfer. To prevent rupture during heterotopic pregnancy, she received laparoscopic surgery to remove the ectopic gestational sac at 7 +2 weeks of gestation. Ultimately, she delivered a healthy boy at 38 +3 weeks of gestation. Here, we also review the clinical presentations, risk factors, treatment options and outcomes of cornual heterotopic pregnancy.

Sandy soils are considered as a significant transition phase to desertification. The effective recovery of sandy soils is of great significance to mitigate the desertification process. Some studies have shown that arbuscular mycorrhizal (AM) fungi and biochar improved the sandy soil, but there have been very few studies regarding the combined effects of AM fungi and biochar amendments on sandy soil improvement. Additionally, the roles of the bacterial and fungal community during the process of sandy soil improvement remain unclear. A greenhouse pot experiment with four treatments, including a control (CK, no amendment), single AM fungi-assisted amendment (RI), single biochar amendment (BC), and combined amendment (BC&#95;RI, biochar plus AM fungi), was set up. This study investigated the effects of different amendment methods on the Nitrariasi birica mycorrhizal colonization, biomass, nutrient (N, P, K, Ca, and Mg) content, soil organic carbon, soil nutrient (TN, TP, and TK) content, and soil water-stable aggregate composition. High throughput sequencing technology was used to investigate the roles of the bacterial and fungal communities during the process of sandy soil improvement. Combined with multiple analysis methods, the improvement mechanisms of different amendment methods were explored. The aim was to provide basic data and scientific basics for reasonably and effectively improving sandy soils. The results indicated that a significant mycorrhiza colonization was observed in the inoculation (RI and BC&#95;RI) treatments, but there was no substantial difference in the mycorrhiza colonization with the RI and BC&#95;RI. Compared with the CK, the shoot biomass and shoot element (N, K, Ca, and Mg) contents were significantly increased in the RI, and the shoot element (N, P, K, Ca, and Mg) contents were significantly increased in the BC and BC&#95;RI; compared with the RI and BC, the root biomass and the root element (P, K, Ca, and Mg) contents were significantly increased in the BC&#95;RI. Compared with the CK, the soil organic carbon contents were significantly increased in the BC and BC&#95;RI, the soil TN contents were significantly increased by 152.54%, and the soil TP and TK contents were significantly decreased by 12.5% and 18.8%, respectively. The proportion of soil aggregates with particle sizes of 0.25-0.05 mm was the highest in each treatment, and the large particle size (>0.25 mm) soil aggregate was significantly increased in the BC&#95;RI. Compared with the CK, the Sobs and Shannon indices of the bacterial/fungal community were significantly decreased in the RI and BC&#95;RI. There was a difference in the microbial community compositions and abundance in the various treatments. The results of the RDA and network analysis were as follows:the effects of AM fungi, biochar, and combined amendment on the soil environment and microbial community structure were significant; in the different amendment treatments, the relationship of the microbial molecular ecological network was significantly changed, and the composition of the core species varied; compared with the RI, there was a higher network connection degree and a richer core species composition in the BC and BC&#95;RI; moreover, the essential role of Rhizophagus intraradices was weaken and the core roles of the other microorganisms (especially bacterial species) were enhanced under the combined effects of biochar and AM fungi. The SEM results demonstrated that the application of AM fungi and biochar could directly affect the bacteria/fungi community structure, and further affect the plant growth and soil properties. The differences in the microbial community structure (especially the change in the microbial interaction) were the key driving factors that led to the difference in the soil improvement effectiveness. In summary, the effects of the different amendment methods on the improvement effectiveness of sandy soils varied. The microbial community played key roles in the process of sandy soil improvement, and there were potential advantages and applications in accelerating the ecological restoration of sandy soils under the combined AM fungi and biochar amendment.

Purpose: The study is aimed at assessing the role of preoperative computerised tomography (CT) examination in the quality of reduction and outcomes in elderly patients with intertrochanteric fracture., Methods: The elderly patients with an intertrochanteric fracture who were treated with proximal femoral nail antirotation were included. The patients were divided into the CT group and the no-CT group according to the presence of preoperative CT examination. Patients' baseline characteristics, quality of reduction, and function were recorded at follow-up. Functional outcomes were evaluated using the Harris hip scores (HHS)., Results: Totally, the study included 182 patients with intertrochanteric fractures, with 85 in the CT group and 97 in the no-CT group, admitted between January 2018 and June 2019. There was no difference in the quality of reduction, HHS, the fracture healing, or postoperative complications between the CT group and the no-CT group. However, the CT group experienced the shorter mean operative time and blood transfusion, compared to the no-CT group., Conclusions: The preoperative CT examination seems to be excessive for elderly patients with an intertrochanteric fracture., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Tao Ma et al.)

Cyfluthrin is a pyrethroid insecticide and common household pesticide. The effect of cyfluthrin on Ca 2+ -related physiology in human osteosarcoma is unclear. This study investigated the effect of cyfluthrin on cytosolic-free Ca 2+ concentrations ([Ca 2+ ] i ) and viability in MG63 human osteosarcoma cells. Cyfluthrin concentration-dependently induced [Ca 2+ ] i rises. Cyfluthrin-induced Ca 2+ entry was confirmed by the Mn 2+ -induced quench of fura-2 fluorescence. Cyfluthrin at concentrations of 10-100 μM induced [Ca 2+ ] i rises. Ca 2+ removal reduced the signal by approximately 50%. Cyfluthrin (100 μM) induced Mn 2+ influx suggesting Ca 2+ entry. Cyfluthrin-induced Ca 2+ entry was inhibited 50% by protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate) and inhibitor (GF109203X) and also by three inhibitors of store-operated Ca 2+ channels: nifedipine, econazole, and SKF96365. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor thapsigargin (TG) completely inhibited cyfluthrin-evoked [Ca 2+ ] i rises. Conversely, treatment with cyfluthrin abolished TG-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C (PLC) with 1-[6-[((17β)-3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dion abolished cyfluthrin-induced [Ca 2+ ] i rises. Cyfluthrin at 25-65 μM decreased cell viability, which was not reversed by pretreatment with the Ca 2+ chelator 1,2-bis(2-aminophenoxy)ethane- N , N , N' , N' -tetraacetic acid-acetoxymethyl ester. Together, in MG63 cells, cyfluthrin induced [Ca 2+ ] i rises by evoking PLC-dependent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via PKC-sensitive store-operated Ca 2+ entry. Cyfluthrin also caused Ca 2+ -independent cell death.

Objective: To observe the clinical therapeutic effect of herb-separated moxibustion at Jinsuo (GV 8)- eight-diagram points on diarrhea-type irritable bowel syndrome (IBS-D) of liver stagnation and spleen deficiency as compared with oral administration of pinaverium bromide tablets and Chinese herbal decoction, tongxieyaofang ., Methods: A total of 126 patients with IBS-D of liver stagnation and spleen deficiency were randomized into a herb-separated moxibustion group (moxibustion group), a western medication group and a Chinese herbal medication group, 42 cases in each one. In the moxibustion group, the herb-separated moxibustion was applied to Jinsuo (GV 8)-eight-diagram points. The herbs in tongxieyaofang (fried rhizoma atractylodis macrocephalae , fried radix paeoniae alba , pericarpium citri reticulatae and radix saposhnikoviae ) were ground into herbal paste and the paste was put on Jinsuo (GV 8)-eight-diagram points. The suspending moxibustion was exerted over the points for 40 min, once daily. In the western medication group, pinaverium bromide tablets were taken orally, 50 mg each time, three times a day. In the Chinese herbal medication group, the decoction of tongxieyaofang was taken orally, one dose a day, taking separately in two times. The duration of treatment was 8 weeks in each group. Before and after treatment, the symptom score of traditional Chinese medicine (TCM), gastrointestinal (GI) symptom score, the score of IBS symptom severity scale (IBS-SSS) and the score of IBS quality of life (IBS-QOL) scale were observed in patients of each group separately. The clinical therapeutic effect was evaluated., Results: After treatment, the scores of TCM symptoms, GI symptom scores and IBS-SSS scores were all obviously reduced in each group ( P <0.05). Each of the scores in the moxibustion group was lower than the western medication group and the Chinese herbal medication group respectively ( P <0.05). After treatment, the scores of each of eight subscale structures of IBS-QOL scale, named dysphoria, interference with activity, body image, health worry, food avoidance, social reaction, sexual intercourse and relationship, were all increased obviously in each group ( P <0.05). The scores of each of eight subscale structures in the moxibustion group were higher than the western medication group and the Chinese herbal medication group respectively ( P <0.05). The total effective rate was 92.9% (39/42) in the moxibustion group, higher than 71.4% (30/42) in the western medication group and 73.8% (31/42) in the Chinese herbal medication group respectively ( P <0.05)., Conclusion: Herb-separated moxibustion at Jinsuo (GV 8)-eight-diagram points remarkably relieves gastrointestinal symptoms and improves the quality of life in patients of diarrhea-type irritable bowel syndrome of liver stagnation and spleen deficiency, and its clinical therapeutic effect is superior to oral administration of either pinaverium bromide tablets or tongxieyaofang .

Thioridazine, belonging to first-generation antipsychotic drugs, is a prescription used to treat schizophrenia. However, the effect of thioridazine on intracellular Ca 2+ concentration ([Ca 2+ ] i ) and viability in human liver cancer cells is unclear. This study examined whether thioridazine altered Ca 2+ signaling and viability in HepG2 human hepatocellular carcinoma cells. Ca 2+ concentrations in suspended cells were measured using the fluorescent Ca 2+ -sensitive dye fura-2. Cell viability was examined by WST-1 assay. Thioridazine at concentrations of 25-100 μM induced [Ca 2+ ] i rises. Ca 2+ removal reduced the signal by 20%. Thioridazine (100 μM) induced Mn 2+ influx suggesting of Ca 2+ entry. Thioridazine-induced Ca 2+ entry was inhibited by 20% by protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate) and inhibitor (GF109203X) and by three inhibitors of store-operated Ca 2+ channels: nifedipine, econazole, and SKF96365. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor thapsigargin (TG) abolished thioridazine-evoked [Ca 2+ ] i rises. On the other hand, thioridazine preincubation completely inhibited the [Ca 2+ ] i rises induced by TG. Furthermore, U73122 totally suppressed the [Ca 2+ ] i rises induced by thioridazine via inhibition of phospholipase C (PLC). Regarding cytotoxicity, at 30-80 μM, thioridazine reduced cell viability in a concentration-dependent fashion. This cytotoxicity was not prevented by preincubation with 1,2-bis (2-aminophenoxy) ethane-N, N, N', N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM) (a Ca 2+ chelator). To conclude, thioridazine caused concentration-dependent [Ca 2+ ] i rises in HepG2 human hepatoma cells by inducing Ca 2+ release from the endoplasmic reticulum via PLC-associated pathways and Ca 2+ influx from extracellular medium through PKC-sensitive store-operated Ca 2+ entry. In addition, thioridazine induced cytotoxicity in a Ca 2+ -independent manner., Competing Interests: None

Tectorigenin, a traditional Chinese medicine, is isolated from the flower of plants such as Pueraria thomsonii Benth. It is an O-methylated isoflavone, a type of flavonoid. Previous studies have shown that tectorigenin evoked various physiological responses in different models, but the effect of tectorigenin on cytosolic-free Ca 2+ levels ([Ca 2+ ] i ) and cytotoxicity in renal tubular cells is unknown. Our research explored if tectorigenin changed Ca 2+ signal transduction and viability in Madin-Darby Canine Kidney (MDCK) renal tubular cells. [Ca 2+ ] i in suspended cells were measured by applying the fluorescent Ca 2+ -sensitive probe fura-2. Viability was explored by using water-soluble tetrazolium-1 as a fluorescent dye. Tectorigenin at concentrations of 5-50 μM induced [Ca 2+ ] i rises. Ca 2+ removal reduced the signal by approximately 20%. Tectorigenin (50 μM) induced Mn 2+ influx suggesting of Ca 2+ entry. Tectorigenin-induced Ca 2+ entry was inhibited by 10% by three inhibitors of store-operated Ca 2+ channels, namely, nifedipine, econazole, and SKF96365. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor thapsigargin inhibited 83% of tectorigenin-evoked [Ca 2+ ] i rises. Conversely, treatment with tectorigenin abolished thapsigargin-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C with U73122 inhibited 50% of tectorigenin-induced [Ca 2+ ] i rises. Tectorigenin at concentrations between 10 and 60 μM killed cells in a concentration-dependent fashion. Chelation of cytosolic Ca 2+ with 1,2-bis (2-aminophenoxy)ethane-N, N, N', N'-tetraacetic acid/acetoxy methyl did not reverse tectorigenin's cytotoxicity. Our data suggest that, in MDCK cells, tectorigenin evoked [Ca 2+ ] i rises and induced cell death that was not associated with [Ca 2+ ] i rises. Therefore, tectorigenin may be a Ca 2+ -independent cytotoxic agent for kidney cells., Competing Interests: None

Sirtuin 3 (SIRT3) modulates mitochondria-localized processes and is implicated in the metabolic reprogramming of cancer cells, especially fatty acid (FA) synthesis. However, the relationship between SIRT3 and aberrant lipid synthesis in cervical cancer remains unclear. Here, we investigated the clinical relevance of SIRT3 expression in cervical squamous cell carcinoma (CSCC), cervical intraepithelial neoplasia (CIN), and normal tissues. To analyze the role of SIRT3 in CCSC in vitro, endogenous SIRT3 levels were up- and down-regulated in SiHa and C33a cells, respectively, via lentiviral-based transfection. Levels were quantified using qRT-PCR. Acetylation levels for acetyl-coA carboxylase (ACC1) were measured with the anti-acetyllysine antibody. Knockdown of SIRT3 reduced levels of cellular lipid content in cells. To investigate the role of SIRT3 in cell proliferation, nude mice were xenografted with SIRT3-overexpressing or SIRT3-knockdown CCSC cells. Overall, the results demonstrate that SIRT3 significantly contributed to the reprogramming of FA synthesis in CCSC by up-regulating ACC1 to promote de novo lipogenesis by SIRT3 deacetylation. Moreover, the findings show that the SIRT3-mediated regulation of FA synthesis played a critical role in the proliferation and metastasis of CCSC cells, suggesting that SIRT3 has therapeutic potential in CCSC treatment.

In recent years, the development of Chinese herbal medicine (CHM) has been challenged by shortages of CHM resources and drug safety concerns related to end products. There have been significant efforts by Chinese scholars to tackle these challenges, which are revealed by analyzing the research trend of CHM resources via surveying Chinese Traditional and Herbal Drugs (Zhong Cao Yao), a representative journal in CHM. Our study focused on 781 articles in CHM resources from 2013 to 2018 and included four subject areas: germplasm resources, quality analysis and evaluation, cultivation, and bioengineering of CHM. Discussion and prospective for future investigations were also presented, including: construct the core germplasm of medicinal plants and expand germplasms; combine molecular research with field experiments and promote the deeper study of cultivation of CHM plants; improve the quality evaluation method of CHM and strengthen the identification of Chinese patented medicines; promote the sustainable development of CHM resources by utilizing bioengineering and synthetic biology. This study helps international scholars understand the status quo of CHM research and provides theoretical support for the healthy, modern, and international development of CHM, and it will facilitate the sustainable development of the traditional Chinese medicine industry., Competing Interests: The authors declare no conflict of interests., (© 2019 Tianjin Press of Chinese Herbal Medicines. Published by Elsevier B.V.)

With the evolution of medical techniques and technology, an increasing number of infants, neonates, and fetuses are exposed to general anesthesia for clinical diagnostic and therapeutic process. The neurotoxic effects of general anesthetics on developing brain have been a subject of concern and considerable research interest. Population-based study confirmed that single short-term general anesthetic exposure does not affect nervous system function, but multiple exposures to general anesthesia could damage cognitive function. Animal studies further discovered the underlying mechanisms. Nervous system is most susceptible to general anesthetics during the brain growth spurt. The time-point is more critical than the duration of exposure to general anesthetics. General anesthetics can induce intracellular calcium overload, disturb energy metabolism, promote cell apoptosis and lead to cell loss. General anesthetics can damage synaptic structure, transmission and plasticity, and impair brain function. High throughput omics technologies have been used to screen the differentially expressed genes induced by general anesthetics, which provide further understanding of the mechanism of general anesthetics affecting cognitive function. This review provides an update on the pathophysiologic mechanisms underlying the anesthesia-neurotoxicity, which will be helpful to provide instructions for the clinical use of general anesthesia in children.

Bifenthrin, a commonly used pyrethroid pesticide, evokes various toxicological effects in different models. However, the effect of bifenthrin on cytosolic-free Ca 2+ level ([Ca 2+ ] i ) and cytotoxicity in human prostate cancer cells is unclear. This study examined whether bifenthrin altered Ca 2+ homeostasis and cell viability in PC3 human prostate cancer cells. [Ca 2+ ] i in suspended cells were measured using the fluorescent Ca 2+ -sensitive dye fura-2. Cell viability was examined by 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 assay. Bifenthrin (100-400 μM) concentration-dependently induced [Ca 2+ ] i rises. Ca 2+ removal reduced the signal by approximately 30%. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) abolished bifenthrin-evoked [Ca 2+ ] i rises. Conversely, treatment with bifenthrin abolished BHQ-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C (PLC) with U73122 significantly inhibited bifenthrin-induced [Ca 2+ ] i rises. Mn 2+ has been shown to enter cells through similar mechanisms as Ca 2+ but quenches fura-2 fluorescence at all excitation wavelengths. Bifenthrin (400 μM)-induced Mn 2+ influx implicates that Ca 2+ entry occurred. Bifenthrin-induced Ca 2+ entry was inhibited by 30% by protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate) and inhibitor (GF109203X) and three inhibitors of store-operated Ca 2+ channels: nifedipine, econazole, and SKF96365. Bifenthrin at 175-275 μM decreased cell viability, which was not reversed by pretreatment with the Ca 2+ chelator 1,2-bis(2-aminophenoxy) ethane- N , N , N ', N '-tetra acetic acid-acetoxymethyl ester. Together, in PC3 cells, bifenthrin-induced [Ca 2+ ] i rises by evoking PLC-dependent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via PKC-sensitive store-operated Ca 2+ entry. Bifenthrin also caused Ca 2+ -independent cell death.

Chlorzoxazone is a skeletal muscle relaxant. However, the effect of chlorzoxazone on intracellular Ca 2+ concentrations ([Ca 2+ ] i ) in oral cancer cells is unclear. This study examined whether chlorzoxazone altered Ca 2+ signaling and cell viability in OC2 human oral cancer cells. [Ca 2+ ] i in suspended cells was measured using the fluorescent Ca 2+ -sensitive dye fura-2. Cell viability was examined by water-soluble tetrazolium-1 assay. Chlorzoxazone (250-1000 μM) induced [Ca 2+ ] i rises in a concentration-dependent manner. Ca 2+ removal reduced the signal by approximately 50%. Mn 2+ has been shown to enter cells through similar mechanisms as Ca 2+ but quenches fura-2 fluorescence at all excitation wavelengths. Chlorzoxazone (1000 μM) induced Mn 2+ influx, suggesting that Ca 2+ entry occurred. Chlorzoxazone-induced Ca 2+ entry was inhibited by 20% by inhibitors of store-operated Ca 2+ channels and protein kinase C (PKC) modulators. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor thapsigargin (TG) inhibited chlorzoxazone-evoked [Ca 2+ ] i rises by 88%. Conversely, treatment with chlorzoxazone-suppressed TG-evoked [Ca 2+ ] i rises 75%. Chlorzoxazone induced [Ca 2+ ] i rises by exclusively releasing Ca 2+ from the endoplasmic reticulum. Inhibition of phospholipase C (PLC) with U73122 did not alter chlorzoxazone-induced [Ca 2+ ] i rises. PLC activity was not involved in chlorzoxazone-evoked [Ca 2+ ] i rises. Chlorzoxazone at 200-700 μM decreased cell viability, which was not reversed by pretreatment with Ca 2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl. In sum, in OC2 cells, chlorzoxazone induced [Ca 2+ ] i rises by evoking PLC-independent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via PKC-sensitive store-operated Ca 2+ entry. Chlorzoxazone also caused Ca 2+ -independent cell death. Since [Ca 2+ ] i rises play a triggering or modulatory role in numerous cellular phenomena, the effect of chlorzoxazone on [Ca 2+ ] i and cell viability should be taken into account in other in vitro studies., Competing Interests: None

Terfenadine, an antihistamine used for the treatment of allergic conditions, affected Ca 2+ -related physiological responses in various models. However, the effect of terfenadine on cytosolic free Ca 2+ levels ([Ca 2+ ] i ) and its related physiology in renal tubular cells is unknown. This study examined whether terfenadine altered Ca 2+ signaling and caused cytotoxicity in Madin-Darby canine kidney (MDCK) renal tubular cells. The Ca 2+ -sensitive fluorescent dye fura-2 was used to measure [Ca 2+ ] i . Cell viability was measured by the fluorescent reagent 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 (WST-1) assay. Terfenadine at concentrations of 100-1000 μM induced [Ca 2+ ] i rises concentration dependently. The response was reduced by approximately 35% by removing extracellular Ca 2+ . In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) partly inhibited terfenadine-evoked [Ca 2+ ] i rises. Conversely, treatment with terfenadine abolished BHQ-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C (PLC) with U73122 inhibited 95% of terfenadine-induced Ca 2+ release. Terfenadine-induced Ca 2+ entry was supported by Mn 2+ -caused quenching of fura-2 fluorescence. Terfenadine-induced Ca 2+ entry was partly inhibited by an activator of protein kinase C (PKC), phorbol 12-myristate 13 acetate (PMA) and by three modulators of store-operated Ca 2+ channels (nifedipine, econazole, and SKF96365). Terfenadine at 200-300 μM decreased cell viability, which was not reversed by pretreatment with the Ca 2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Together, in MDCK cells, terfenadine induced [Ca 2+ ] i rises by evoking PLC-dependent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via PKC-sensitive store-operated Ca 2+ entry. Furthermore, terfenadine caused cell death that was not triggered by preceding [Ca 2+ ] i rises.

Timolol is a medication used widely to treat glaucoma. Regarding Ca 2+ signaling, timolol was shown to modulate Ca 2+ -related physiology in various cell types, however, the effect of timolol on Ca 2+ homeostasis and cell viability has not been explored in human prostate cancer cells. The aim of this study was to explore the effect of timolol on intracellular Ca 2+ concentrations ([Ca 2+ ] i ) and viability in PC3 human prostate cancer cells. Timolol at concentrations of 100-1000 μM induced [Ca 2+ ] i rises. The Ca 2+ signal in Ca 2+ -containing medium was reduced by removal of extracellular Ca 2+ by approximately 75%. Timolol (1000 μM) induced Mn 2+ influx suggesting of Ca 2+ entry. Timolol-induced Ca 2+ entry was partially inhibited by three inhibitors of store-operated Ca 2+ channels: nifedipine, econoazole and SKF96365, and by a protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate [PMA]) or an inhibitor (GF109203X). In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor thapsigargin abolished timolol-evoked [Ca 2+ ] i rises. Conversely, treatment with timolol abolished thapsigargin-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C (PLC) with U73122 abolished timolol-induced [Ca 2+ ] i rises. Timolol at concentrations between 200 and 600 μM killed cells in a concentration-dependent fashion. Chelation of cytosolic Ca 2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM (BAPTA/AM) did not reverse cytotoxicity of timolol. Together, in PC3 cells, timolol induced [Ca 2+ ] i rises by evoking Ca 2+ release from the endoplasmic reticulum in a PLC-dependent manner, and Ca 2+ influx via PKC-regulated store-operated Ca 2+ entry. Timolol also caused cell death that was not linked to preceding [Ca 2+ ] i rises.

Niflumic acid, a drug used for joint and muscular pain, affected Ca²⁺ signaling in different models. However, the effect of niflumic acid on Ca²⁺ homeostasis and Ca²⁺-related physiology in human osteosarcoma cells is unknown. This study examined the effect of niflumic acid on cytosolic free Ca²⁺ concentrations ([Ca²⁺]i) in MG63 human osteosarcoma cells. Intracellular Ca²⁺ concentrations in suspended cells were monitored by using the fluorescent Ca²⁺-sensitive dye fura- 2. Cell viability was examined by using 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio- 1,3-benzene disulfonate] water soluble tetrazolium-1 (WST-1). In MG63 cells, niflumic acid at concentrations of 250-750 μM evoked [Ca²⁺]i rises concentration-dependently. Niflumic acid-evoked Ca²⁺ entry was confirmed by Mn²⁺-induced quenching of fura-2 fluorescence. This entry was inhibited by nifedipine, econazole, SKF96365, the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate (PMA), but was not affected by the PKC inhibitor GF109203X. In Ca²⁺- free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin (TG) inhibited niflumic acid-evoked [Ca²⁺]i rises. Conversely, treatment with niflumic acid abolished TG-evoked [Ca²⁺]i rises. Inhibition of phospholipase C (PLC) with U73122 also partly reduced niflumic acid-evoked [Ca²⁺]i rises. Niflumic acid killed cells at 200-500 μM in a concentration-dependent fashion. Chelating cytosolic Ca²⁺ with 1,2-bis(2-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid/ AM (BAPTA/AM) did not reverse niflumic acid-induced cytotoxicity. Collectively, our data suggest that in MG63 cells, niflumic acid induced [Ca²⁺]i rises by evoking PLC-dependent Ca²⁺ release from the endoplasmic reticulum, and Ca²⁺ entry via PKC-sensitive store-operated Ca²⁺ entry. Niflumic acid also induced Ca²⁺-independent cell death., Competing Interests: The authors declared no conflicts of interest.

Thirteen previously undescribed lycodine-type Lycopodium alkaloids, namely, five alkaloids (lycocasuarines D-H) each possessing an uncommon five-membered C ring and eight Lycopodium alkaloid glycosides (casuarinosides A-H), together with a known analog, were isolated from the aerial parts of Lycopodiastrum casuarinoides (Spring) Holub ex R.D.Dixit (Lycopodiaceae). The structures of the compounds were elucidated by extensive spectroscopic analysis, single-crystal X-ray diffraction, and chemical methods. In addition, the acetylcholinesterase inhibitory activity of the isolated compounds was evaluated., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Captopril, an angiotensin-converting enzyme (ACE) inhibitor, induced different Ca²⁺ signaling responses in various cell models. However, the effect of captopril on Ca²⁺ homeostasis and cell viability in hepatoma cells is unknown. This study examined whether captopril altered Ca²⁺ homeostasis and viability in HepG2 human hepatoma cells. Intracellular Ca²⁺ concentrations in suspended cells were monitored by using the fluorescent Ca²⁺-sensitive dye fura-2. Cell viability was examined by using 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 (WST-1). Captopril at concentrations of 500-3000 μM induced [Ca²⁺]i rises in a concentration-dependent manner. Ca²⁺ removal reduced the signal by approximately 15%. Mn²⁺ has been shown to enter cells through similar mechanisms as Ca²⁺ but quenches fura-2 fluorescence at all excitation wavelengths. Captopril (3000 μM)-induced Mn²⁺ influx indirectly suggested that captopril evoked Ca²⁺ entry. Captopril-induced Ca²⁺ entry was inhibited by 15% by a protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate, PMA) and an inhibitor (GF109203X) and three inhibitors of store-operated Ca²⁺ channels: nifedipine, econazole and SKF96365. In Ca²⁺-free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) abolished captopril-evoked [Ca²⁺]i rises. Conversely, treatment with captopril abolished BHQ-evoked [Ca²⁺]i rises. Inhibition of phospholipase C (PLC) with U73122 inhibited 70% of captopril-induced [Ca²⁺]i rises. Captopril at concentrations between 150-550 μM killed cells in a concentration-dependent fashion. Chelation of cytosolic Ca²⁺ with 1,2-bis(2-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid/AM (BAPTA/AM) did not reverse captopril’s cytotoxicity. Together, in HepG2 human hepatoma cells, captopril induced [Ca²⁺]i rises and caused cell death that was not triggered by preceding [Ca²⁺]i rises., Competing Interests: The authors declare that there are no conflicts of interests.

Objective: Magnolol, a polyphenol compound from herbal medicines, was shown to alter physiology in various cell models. However, the effect of magnolol on Ca 2+ homeostasis and its related physiology in oral cancer cells is unclear. This study examined whether magnolol altered Ca 2+ signaling and cell viability in OC2 human oral cancer cells., Methods: Cytosolic Ca 2+ concentrations ([Ca 2+ ] i ) in suspended cells were measured by using the fluorescent Ca 2+ -sensitive dye fura-2. Cell viability was examined by 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 (WST-1) assay., Results: Magnolol at concentrations of 20-100 μM induced [Ca 2+ ] i rises. Ca 2+ removal reduced the signal by approximately 50%. Magnolol (100 μM) induced Mn 2+ influx suggesting of Ca 2+ entry. Magnolol-induced Ca 2+ entry was partially suppressed by protein kinase C (PKC) regulators, and inhibitors of store-operated Ca 2+ channels. In Ca 2+ -free medium, treatment with the endoplasmic reticulum Ca 2+ pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) abolished magnolol-evoked [Ca 2+ ] i rises. Conversely, treatment with magnolol abolished BHQ-evoked [Ca 2+ ] i rises. Inhibition of phospholipase C (PLC) with U73122 partially inhibited magnolol-induced [Ca 2+ ] i rises. Magnolol at 20-100 μM decreased cell viability, which was not reversed by pretreatment with the Ca 2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM)., Conclusions: Together, in OC2 cells, magnolol induced [Ca 2+ ] i rises by evoking partially PLC-dependent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via PKC-sensitive store-operated Ca 2+ entry. Magnolol also caused Ca 2+ -independent cell death. Therefore, magnolol-induced cytotoxicity may not be involved in activation mechanisms associated with intracellular Ca 2+ mobilization in oral cancer cells., (Copyright © 2018. Published by Elsevier Ltd.)

Sugars Will Eventually be Exported Transporter (SWEET) and SemiSWEET are recently characterized families of sugar transporters in eukaryotes and prokaryotes, respectively. SemiSWEETs contain 3 transmembrane helices (TMHs), while SWEETs contain 7. Here, we performed sequence-based comprehensive analyses for SWEETs and SemiSWEETs across the biosphere. In total, 3,249 proteins were identified and ≈60% proteins were found in green plants and Oomycota, which include a number of important plant pathogens. Protein sequence similarity networks indicate that proteins from different organisms are significantly clustered. Of note, SemiSWEETs with 3 or 4 TMHs that may fuse to SWEET were identified in plant genomes. 7-TMH SWEETs were found in bacteria, implying that SemiSWEET can be fused directly in prokaryote. 15-TMH extraSWEET and 25-TMH superSWEET were also observed in wild rice and oomycetes, respectively. The transporters can be classified into 4, 2, 2, and 2 clades in plants, Metazoa, unicellular eukaryotes, and prokaryotes, respectively. The consensus and coevolution of amino acids in SWEETs were identified by multiple sequence alignments. The functions of the highly conserved residues were analyzed by molecular dynamics analysis. The 19 most highly conserved residues in the SWEETs were further confirmed by point mutagenesis using SWEET1 from Arabidopsis thaliana . The results proved that the conserved residues located in the extrafacial gate (Y57, G58, G131, and P191), the substrate binding pocket (N73, N192, and W176), and the intrafacial gate (P43, Y83, F87, P145, M161, P162, and Q202) play important roles for substrate recognition and transport processes. Taken together, our analyses provide a foundation for understanding the diversity, classification, and evolution of SWEETs and SemiSWEETs using large-scale sequence analysis and further show that gene duplication and gene fusion are important factors driving the evolution of SWEETs.

c-Met/HGF signaling pathway plays an important role in cancer progression, and it was considered to be related to poor prognosis and drug resistance. Based on metabolite profiling of (S)-7-fluoro-6-(1-(6-(1-methyl-1H-pyrazol-4-yl)-1H-imidazo[4,5-b]pyrazin-1-yl)ethyl)quinoline (1), a series of 2-substituted or 3-substituted-6-(1-(1H-[1,2,3]triazolo[4,5-b]pyrazin-1-yl)ethyl)quinoline derivatives was rationally designed and evaluated. Most of the 3-substituted derivatives not only exhibited potent activities in both enzymatic and cellular assays, but also were stable in liver microsomes among different species (human, rat and monkey). SAR investigation revealed that introducing of N-methyl-1H-pyrazol-4-yl group at the 3-position of quinoline moiety is beneficial to improve the inhibitory potency, especially in the cellular assays. The influence of fluorine atom at 7-position or 5, 7-position of quinoline moiety and substituents at the 6-position of triazolo[4,5-b]pyrazine core on overall activity is not very significant. Racemate 14, an extremely potent and exquisitely selective c-Met inhibitor, demonstrated favorable pharmacokinetic properties in rats, no significant AO metabolism, and effective tumor growth inhibition in c-Met overexpressed NSCLC (H1993 cell line) and gastric cancer (SNU-5 cell line) xenograft models. Docking analysis indicated that besides the typical interactions of most selective c-Met inhibitors, the intramolecular halogen bond and additional hydrogen bond interactions with kinase are beneficial to the binding. These results may provide deep insight into potential structural modifications for developing potent c-Met inhibitors., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Objective: To investigate the change in the expression of tight junction protein ZO-1 in intestinal epithelial cells (Caco-2 cells) and the protective effect of eicosapentaenoic acid (EPA) after adherent-invasive Escherichia coli (E.coli) LF82 infection., Methods: The Caco-2 cell line was used to establish an in vitro model of tight junction of intestinal epithelial cells. Caco-2 cells were divided into EPA treatment groups (0, 25, 50, 100, and 200 μmol/L EPA) and EPA (0, 25, 50, 100, and 200 μmol/L EPA)+E.coli LF82 treatment (0, 6, and 12 hours) groups. A microscope was used to observe the morphological characteristics of the cells. MTT assay was used to determine the cell growth curve. The activity of alkaline phosphatase (ALP) at both sides of the cell membrane was compared to evaluate the Caco-2 cell model. MTT assay and flow cytometry were used to investigate the effects of different concentrations of EPA on the survival rate and apoptosis rate of Caco-2 cells. RT-qPCR was used to measure the mRNA expression of ZO-1 in Caco-2 cells after EPA and/or E.coli LF82 treatment. ELISA was used to measure the change in the level of tumor necrosis factor-α (TNF-α) in culture supernatant., Results: After EPA treatment (25 and 50 μmol/L), the proliferation of Caco-2 cells was induced in a dose-dependent manner. The survival rates of the cells were significantly higher than those in the control group (P<0.05). The EPA treatment (100 and 200 μmol/L) groups had a significant inhibitory effect on the proliferation of Caco-2 cells in a dose-dependent manner. The survival rates of the cells were significantly lower than those in the control group (P<0.05). The EPA treatment (100 and 200 μmol/L) groups had a significant increase in cell apoptosis rate compared with the control group (P<0.05). The 6- and 12-hour E.coli LF82 treatment groups had decreasing mRNA expression of ZO-1 in Caco-2 cells over the time of treatment and had significantly lower mRNA expression of ZO-1 than the untreated group (P<0.05). The Caco-2 cells treated with E.coli LF82 and 25 or 50 μmol/L EPA for 6 or 12 hours showed an increase in the mRNA expression of ZO-1 with the increasing concentration of EPA, as well as significantly higher mRNA expression of ZO-1 than the Caco-2 cells treated with E.coli LF82 alone (P<0.05). The Caco-2 cells treated with E.coli LF82 alone for 6 or 12 hours had increasing secretion of TNF-α over the time of treatment and had significantly higher secretion than the untreated Caco-2 cells (P<0.05). The Caco-2 cells treated with E.coli LF82 and 25 or 50 μmol/L EPA for 6 or 12 hours showed a reduction in the secretion of TNF-α with the increasing concentration of EPA and had significantly lower secretion than the Caco-2 cells treated with E.coli LF82 alone (P<0.05)., Conclusions: EPA can effectively prevent the destruction of tight junction of intestinal epithelial cells induced by E.coli LF82 infection and inhibit the secretion of inflammatory factors. Therefore, it has a certain protective effect on intestinal mucosal barrier.

Accumulating evidence suggests that an abnormal accumulation of iron in the substantia nigra (SN) is one of the defining characteristics of Parkinson's disease (PD). Accordingly, the potential neuroprotection of Fe chelators is widely acknowledged for the treatment of PD. Although desferrioxamine (DFO), an iron chelator widely used in clinical settings, has been reported to improve motor deficits and dopaminergic neuronal survival in animal models of PD, DFO has poor penetration to cross the blood-brain barrier and elicits side effects. We evaluated whether an intranasal administration of DFO improves the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced degeneration of dopaminergic neurons in the nigrostriatal axis and investigated the molecular mechanisms of intranasal DFO treatment in preventing MPTP-induced neurodegeneration. Treatment with DFO efficiently alleviated behavioral deficits, increased the survival of tyrosine hydroxylase (TH)-positive neurons, and decreased the action of astrocytes in the SN and striatum in an MPTP-induced PD mouse model. Interestingly, we found that DFO up-regulated the expression of HIF-1α protein, TH, vascular endothelial growth factor (VEGF), and growth associated protein 43 (GAP43) and down-regulated the expression of α-synuclein, divalent metal transporter with iron-responsive element (DMT1+IRE), and transferrin receptor (TFR). This was accompanied by a decrease in iron-positive cells in the SN and striatum of the DFO-treated group. We further revealed that DFO treatment significantly inhibited the MPTP-induced phosphorylation of the c-Jun N-terminal kinase (JNK) and differentially enhanced the phosphorylation of extracellular regulated protein kinases (ERK) and mitogen-activated protein kinase (MAPK)/P38 kinase. Additionally, the effects of DFO on increasing the Bcl-2/Bax ratio were further validated in vitro and in vivo. In SH-SY5Y cells, the DFO-mediated up-regulation of HIF-1α occurred via the activation of the ERK and P38MAPK signaling pathway. Collectively, the present data suggest that intranasal DFO treatment is effective in reversing MPTP-induced brain abnormalities and that HIF-1-pathway activation is a potential therapy target for the attenuation of neurodegeneration., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: To investigate the expression of LRG-1 in clinical specimens and Tca8113 cell line of tongue carcinoma and analyze the relationship between LRG-1 expression and the clinicopathological parameters., Methods: LRG-1 expression was detected in 40 tongue squamous cell carcinoma (TSCC) tissues and paired normal adjacent tissues, 20 atypical hyperplasia tissues of the tongue, and 20 tissues of tongue cancer in situ using immunohistochemical method. The expression of LRG-1 in Tca8113 cell line was detected using flow cytometry. The expression of LRG-1 was also detected in human TSCC tissues and Tca8113 cells with Western blotting. The effect of LRG-1 on the proliferation of HUVECs was determined using MTT assay, and its effect on angiogenesis was evaluated with Matrigel tube formation assays., Results: Human TSCC tissues had a significantly higher rate of positive expression for LRG-1 (85%, 34/40) than the adjacent tissues (10%, 4/40), invasive tongue cancer (30%, 6/20), and tongue cancer in situ (50%, 10/20) (P<0.05). LRG-1 expression was correlated with the degree of tumor differentiation, clinical stage and lymph node metastasis of the tumor (P<0.05) but not with the patients' age or gender. In the in vitro experiment, LRG-1 promoted HUVEC proliferation and angiogenesis., Conclusion: Abnormal LRG-1 expression is present in the human TSCC tissue and Tca8113 cells. LRG-1 can promote HUVEC proliferation and angiogenesis in vitro, suggesting its possible role in promoting tumor angiogenesis.