Mitigating phospholipid peroxidation of macrophages in stress-induced tumor microenvironment by natural ALOX15/PEBP1 complex inhibitors.

BACKGROUND: The intricate interactions between chronic psychological stress and susceptibility to breast cancer have been recognized, yet the underlying mechanisms remain unexplored. Danzhi Xiaoyao Powder (DZXY), a traditional Chinese medicine (TCM) formula, has found clinical utility in the treatment of breast cancer. Macrophages, as the predominant immune cell population within the tumor microenvironment (TME), play a pivotal role in orchestrating tumor immunosurveillance. Emerging evidence suggests that lipid oxidation accumulation in TME macrophages, plays a critical role in breast cancer development and progression. However, a comprehensive understanding of the pharmacological mechanisms and active components of DZXY related to its clinical application in the treatment of stress-aggravated breast cancer remains elusive. PURPOSE: This study sought to explore the plausible regulatory mechanisms and identify the key active components of DZXY contributing to its therapeutic efficacy in the context of breast cancer. METHODS: Initially, we conducted an investigation into the relationship between the phagocytic capacity of macrophages damaged by psychological stress and phospholipid peroxidation using flow cytometry and LC-MS/MS-based phospholipomics. Subsequently, we evaluated the therapeutic efficacy of DZXY based on the results of the tumor size, tumor weight, the phospholipid peroxidation pathway and phagocytosis of macrophage. Additionally, the target-mediated characterization strategy based on binding of arachidonate 15-lipoxygenase (ALOX15) to phosphatidylethanolamine-binding protein-1 (PEBP1), including molecular docking analysis, microscale thermophoresis (MST) assay, co-immunoprecipitation analysis and activity verification, has been further implemented to reveal the key bio-active components in DZXY. Finally, we evaluated the therapeutic efficacy of isochlorogenic acid C (ICAC) based on the results of tumor size, tumor weight, the phospholipid peroxidation pathway, and macrophage phagocytosis in vivo. RESULTS: The present study demonstrated that phospholipid peroxides, as determined by LC-MS/MS-based phospholipidomics, triggered in macrophages, which in turn compromised their capacity to eliminate tumor cells through phagocytosis. Furthermore, we elucidate the mechanism behind stress-induced PEBP1 to form a complex with ALOX15, thereby mediating membrane phospholipid peroxidation in macrophages. DZXY, demonstrates potent anti-breast cancer therapeutic effects by disrupting the ALOX15/PEBP1 interaction and inhibiting phospholipid peroxidation, ultimately enhancing macrophages' phagocytic capability towards tumor cells. Notably, ICAC emerged as a promising active component in DZXY, which can inhibit the ALOX15/PEBP1 interaction, thereby mitigating phospholipid peroxidation in macrophages. CONCLUSION: Collectively, our findings elucidate stress increases the susceptibility of breast cancer by driving lipid peroxidation of macrophages and suggest the ALOX15/PEBP1 complex as a promising intervention target for DZXY.

Babao Dan decreases hepatocarcinogenesis by inhibiting hepatic progenitor cells malignant transformation via down-regulating toll-like receptor 4.

Background: Babao Dan (BBD) is a traditional Chinese medicine that has been widely used as a complementary and alternative medicine to treat chronic liver diseases. In this study, we aimed to observe the effect of BBD on the incidence of diethylnitrosamine (DEN)-initiated hepatocellular carcinoma formation in rats and explored its possible mechanism. Methods: To verify this hypothesis, BBD was administrated to rats at a dose of 0.5g/kg body weight per two days from the 9th to 12th week in HCC-induced by DEN. Liver injury biomarkers and hepatic inflammatory parameters were evaluated by histopathology as well as serum and hepatic content analysis. We applied immunohistochemical analysis to investigate the expression of CK-19 and SOX-9 in liver tissues. The expression of TLR4 was determined by immunohistochemical, RT-PCR, and western blot analysis. Furthermore, we also detected the efficacy of BBD against primary HPCs neoplastic transformation induced by LPS. Results: We observed that DEN could induce hepatocarcinogenesis, and BBD could obviously decrease the incidence. The biochemical and histopathological examination results confirmed that BBD could protect against liver injury and decrease inflammatory infiltration. Immunohistochemistry staining results showed that BBD could effectively inhibit the ductal reaction and the expression of TLR4. The results showed that BBD-serumcould obviously inhibit primary HPCs neoplastic transformation induced by regulating the TLR4/Ras/ERK signaling pathway. Conclusion: In summary, our results indicate that BBD has potential applications in the prevention and treatment of HCC, which may be related to its effect on hepatic progenitor cells malignant transformation via inhibiting the TLR4/Ras/ERK signaling pathway.

Towards advanced simultaneous nitrogen removal and phosphorus recovery from digestion effluent based on anammox-hydroxyapatite (HAP) process: Focusing on a solution perspective.

In this paper, the state-of-the-art information on the anammox-HAP process is summarized. The mechanism of this process is systematically expounded, the enhancement of anammox retention by HAP precipitation and the upgrade of phosphorus recovery by anammox process are clarified. However, this process still faces several challenges, especially how to deal with the âˆ¼ 11% nitrogen residues and to purify the recovered HAP. For the first time, an anaerobic fermentation (AF) combined with partial denitrification (PD) and anammox-HAP (AF-PD-Anammox-HAP) process is proposed to overcome the challenges. By AF of the organic impurities of the anammox-HAP granular sludge, organic acid is produced to be used as carbon source for PD to remove the nitrogen residues. Simultaneously, pH of the solution drops, which promotes the dissolution of some inorganic purities such as CaCO3. In this way, not only the inorganic impurities are removed, but the inorganic carbon is supplied for anammox bacteria.

Mapping Pu'er tea plantations from GF-1 images using Object-Oriented Image Analysis (OOIA) and Support Vector Machine (SVM).

Tea is the most popular drink worldwide, and China is the largest producer of tea. Therefore, tea is an important commercial crop in China, playing a significant role in domestic and foreign markets. It is necessary to make accurate and timely maps of the distribution of tea plantation areas for plantation management and decision making. In the present study, we propose a novel mapping method to map tea plantation. The town of Menghai in the Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, China, was chosen as the study area, andgg GF-1 remotely sensed data from 2014-2017 were chosen as the data source. Image texture, spectral and geometrical features were integrated, while feature space was built by SEparability and THresholds algorithms (SEaTH) with decorrelation. Object-Oriented Image Analysis (OOIA) with a Support Vector Machine (SVM) algorithm was utilized to map tea plantation areas. The overall accuracy and Kappa coefficient ofh the proposed method were 93.14% and 0.81, respectively, 3.61% and 0.05, 6.99% and 0.14, 6.44% and 0.16 better than the results of CART method, Maximum likelihood method and CNN based method. The tea plantation area increased by 4,095.36 acre from 2014 to 2017, while the fastest-growing period is 2015 to 2016.

Combined ultrasonography and CT for prognosis and predicting clinical outcomes of patients with pseudomyxoma peritonei.

OBJECTIVES: This study aimed to identify the diagnostic accuracy of combined ultrasonography (US) and computed tomography (CT) in evaluating the tumor burden of pseudomyxoma peritonei (PMP). Besides, we assessed the ability of this combination to predict the likelihood of complete resection. METHODS: This retrospective study involved 504 patients diagnosed with PMP and scheduled for cytoreduction surgery. We compared tumor burden-quantified as peritoneal cancer index (PCI) by preoperative US and CT (US-CT-PCI)-with surgical findings. Next, we assessed the prognostic value of US-CT PCI and imaging features in determining the completeness of cytoreduction (CCR) score using multivariate analysis. RESULTS: US-CT PCI demonstrated a high PCI evaluation accuracy under moderate tumor burden. Higher US-CT PCI could predict incomplete resection. In addition, we identified imaging features such as mesenteric involvement as an independent predictor of incomplete resection (hazard ratio (HR) = 2.006; p = 0.007). CONCLUSIONS: US-CT PCI allowed us to predict the completeness of cytoreductive surgery in patients with PMP. Moreover, the combined US and CT imaging detected several features indicating incomplete cytoreduction. KEY POINTS: • Ultrasonography (US) can act as a complementary diagnostic modality in peritoneal cancer index (PCI) evaluation by combining CT in the small bowel area and US in the abdominal area. • A modified peritoneal cancer index (US-CT PCI) helps preoperatively evaluate tumor burden with high accuracy and allows to predict incomplete resection. • US-CT PCI of 20 or above and the involvement of particular structures such as mesentery, independently indicate incomplete resection.

[Rauwolfia extract inhibits the proliferation of prostate cells in rats with benign prostatic hyperplasia].

OBJECTIVE: To investigate the effects of different concentrations of Rauwolfia extract (RE) on the proliferation of prostate cells in the rat model of benign prostatic hyperplasia (BPH). METHODS: We randomly divided 48 male SD rats into six groups of an equal number, BPH model control, finasteride, low-concentration RE, medium-concentration RE, high-concentration RE and normal control, and established a BPH model in the former five groups by subcutaneous injection of testosterone propionate following castration. We treated the rats of the finasteride and RE groups intragastrically with finasteride solution at 5 mg/kg and RE at 5, 10 and 20 mg/kg respectively, and those of the model control and normal control groups with an equal dose of normal saline, all once a day for 28 consecutive days. Then, we killed all the animals, collected their prostate tissue, obtained the wet weight and volume of the prostate, the prostate index and the contents of serum T and dihydrotestosterone (DHT), observed the morphological changes of the prostate tissue by HE staining, counted the glands in the prostate tissue, measured the intraglandular area, and determined the expressions of PCNA and α-SMA by immunohistochemistry. RESULTS: Compared with the rats of the normal control group, the BPH model controls showed significantly increased wet weight (ï¼»0.923 ± 0.15ï¼½ vs ï¼»1.455 ± 0.52ï¼½ g, P < 0.05), volume (ï¼»1.035 ± 0.29ï¼½ vs ï¼»1.687 ± 0.31ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.23 ± 0.04ï¼½% vs ï¼»0.37 ± 0.15ï¼½%, P < 0.05), dilation, hyperemia and edema of the prostatic stroma and vessels, and proliferation rate of the prostatic cells, but remarkably decreased number of glands (ï¼»20.35 ± 3.83ï¼½ vs ï¼»12.56 ± 2.58ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.52 ± 1.52ï¼½ µm, P < 0.05) and intraglandular area (ï¼»12.3 ± 1.21ï¼½ vs ï¼»5.96 ± 0.34ï¼½ ×103µm2, P < 0.05). In comparison with the BPH model controls, the animals treated with RE, especially in the high-concentration RE group, exhibited marked decreases in the weight (ï¼»1.455 ± 0.52ï¼½ vs ï¼»0.862 ± 0.31ï¼½ g, P < 0.05), volume ( ï¼»1.687 ± 0.31ï¼½ vs ï¼»0.952 ± 0.28ï¼½ ml, P < 0.05) and index of the prostate (ï¼»0.37 ± 0.15ï¼½% vs ï¼»0.22 ± 0.07ï¼½%, P < 0.05), dramatic improvement in the number of glands (ï¼»12.56 ± 2.58ï¼½ vs ï¼»18.36 ± 1.25ï¼½, P < 0.05), epithelial thickness (ï¼»39.76 ± 5.20ï¼½ vs ï¼»19.04 ± 3.89ï¼½ µm, P < 0.05) and intraglandular area (ï¼»5.96 ± 0.34ï¼½ vs ï¼»10.25 ± 0.98ï¼½ ×103µm2, P<0.05ï¼½, P < 0.05), remarkable down-regulation of the expressions of PCNA and α-SMA, and significant reduction of the contents of serum T (ï¼»19.147 ± 3.214ï¼½ vs ï¼»6.016 ± 1.978ï¼½ ng/ml, P < 0.05) and DHT (ï¼»9.052 ± 0.633ï¼½ vs ï¼»2.532 ± 0.386ï¼½ ng/ml, P < 0.05). CONCLUSION: Rauwolfia extract can inhibit the proliferation of prostate cells and relieve BPH symptoms in a concentration-dependent manner in rats with BPH.

Preparation, characterization, and property evaluation of Hericium erinaceus peptide-calcium chelate.

Recently, owing to the good calcium bioavailability, peptide-calcium chelates made of various foods have been emerging. Hericium erinaceus, an edible fungus, is rich in proteins with a high proportion of calcium-binding amino acids. Thus, mushrooms serve as a good source to prepare peptide-calcium chelates. Herein, the conditions for hydrolyzing Hericium erinaceus peptides (HP) with a good calcium-binding rate (CBR) were investigated, followed by the optimization of HP-calcium chelate (HP-Ca) preparation. Furthermore, the structure of the new chelates was characterized along with the evaluation of gastrointestinal stability and calcium absorption. Papain and a hydrolysis time of 2 h were selected for preparing Hericium erinaceus peptides, and the conditions (pH 8.5, temperature 55°C, time 40 min, and mass ratio of peptide/CaCl2 4:1) were optimal to prepare HP-Ca. Under this condition, the chelates contained 6.79 ± 0.13% of calcium. The morphology and energy disperse spectroscopy (EDS) analysis showed that HP-Ca was loose and porous, with an obvious calcium element signal. The ultraviolet-visible (UV) absorption and Fourier transform infrared spectroscopy (FT-IR) analysis indicated that calcium possibly chelates to HP via interaction with free -COO- from acidic amino acids and C = O from amide. HP-Ca displayed good stability against stimulated gastrointestinal digestion. Moreover, HP-Ca significantly improved the calcium absorption by Caco-2 epithelial cells. Thus, HP-Ca is a promising Ca supplement with high calcium bioavailability.

Antioxidant Properties of Triterpenoids Isolated from Bagasse-Cultivated Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), at Different Developmental Stages.

Although triterpenoids are one of the main active ingredients in Ganoderma lucidum, their accumulation and antioxidant activity during the different developmental stages of G. lucidum cultivation in bagasse remains unclear. In this study, we investigated the content and antioxidant activity of total triterpenoids extracted from G. lucidum strain GL102 during the five growth stages. The obtained results showed that the highest content (12.06 mg/g) was detected in stage 3 (young fruiting body), similar to the contents of ganoderic acids B and G. However, ganoderic acids A and D exhibited maximal contents in stage 5 (spore-ejected fruiting body). The triterpenoids extracted during stage 5 were most capable of scavenging DPPH, OH, and ABTS(+) radicals, with scavenging rates of 65.88%, 86.45%, and 97.91%, respectively. Based on in vivo antioxidant assays conducted on zebrafish, the safe concentration of these triterpenoids was 0.03 mg/mL. At this concentration, the G. lucidum triterpenoids extracted during stage 5 could decrease lipopolysaccharide-induced intracellular reactive oxygen species to a level that was nearly normal (similar to the control group). The accumulation profile and antioxidant activity results reported herein provide the scientific basis needed to promote the utilization of triterpenoids derived from bagasse-cultivated G. lucidum.

Granular activated carbon promoting re-granulation of anammox-hydroxyapatite granules for stable nitrogen removal at low phosphate concentration.

Anaerobic ammonium oxidation (anammox) coupled with hydroxyapatite (HAP) crystallization not only achieves simultaneous nitrogen removal and phosphorus recovery, but also cultivates excellent anammox granules. However, a floatation and wash-out of anammox-HAP granules was occurred at low phosphate concentrations. In this study, a reactor inoculated with mature anammox-HAP granules and fed with low phosphate (5 mg P/L) was added with granular activated carbon (GAC) to maintain sludge granulation and nitrogen-removing stability. At influent total nitrogen >800 mg/L and nitrogen loading rate ~ 9.8 kg/m3/d, a satisfactory nitrogen removal of around 88% was maintained during 140 days of operation. Insufficient phosphate supplement resulted in a sludge bulking, with suspended solid and sludge density decreased whereas sludge water content and expansion ratio increased due to HAP loss. Nevertheless, the sludge re-granulation was found at the later stage as the proportion of granules in 2.8- 3.35 mm went up to 37.4% after large granules disintegrated into small pieces at the initial stage. The settling velocity was finally ranging from 129.8 to 182.2 m/h. In addition, Candidatus Brocadia was increased from 2.1% to 20.1% and dominated in the microbial community. These findings suggest GAC was able to promote re-granulation of anammox-HAP granules at low phosphate concentration, which avoids sludge flotation and widens their application as an inoculum.

[Research progress of Gan-Yu-Hua-Huo syndrome based on emotional stress-induced latent herpes simplex virus-1 reactivation].

Gan-Yu-Hua-Huo syndrome(Live qi stagnation transforming into fire pattern) is one of the core contents of the theory of emotional diseases in traditional Chinese medicine(TCM). It is the key link of the pathogenesis change of emotion-related diseases and widely exists in the pathological process of various related diseases. However, due to the lack of animal models in line with the characteristics of TCM syndromes, the research on biomedical basis of Gan-Yu-Hua-Huo syndrome and study of Chinese medicines for soothing liver and purging fire have been restricted seriously. This study found that the pathological process of facial fire-heat symptoms of Gan-Yu-Hua-Huo syndrome was similar to the facial symptoms due to the emotional stress-induced latent herpes simplex virus-1(HSV-1) reactivation. Therefore, this study proposed that the emotional stress-induced latent HSV-1 activation be used to establish the animal model of Gan-Yu-Hua-Huo syndrome. In this study, the state-of-art literature in the field of Gan-Yu-Hua-Huo syndrome was summarized, and the experimental animal model of Gan-Yu-Hua-Huo syndrome was established from the perspective of emotional stress-induced latent HSV-1 reactivation to reveal the active substances, potential targets and pathways related to the pathological mechanism of the syndrome. This study was expected to provide reference and basis for the pharmacodynamic characterization of commonly used Chinese medicine for Gan-Yu-Hua-Huo syndrome in clinical practice.

Theacrine, a Potent Antidepressant Purine Alkaloid from a Special Chinese Tea, Promotes Adult Hippocampal Neurogenesis in Stressed Mice.

Daily intake of tea has been known to relate to a low risk of depression. In this study, we report that a special variety of tea in China, Camellia assamica var. kucha (kucha), possesses antidepressant effects but with less adverse effects as compared to traditional tea Camellia sinensis. This action of kucha is related to its high amount of theacrine, a purine alkaloid structurally similar to caffeine. We investigated the antidepressant-like effects and mechanisms of theacrine in chronic water immersion restraint stress and chronic unpredictable mild stress mice models. PC12 cells and primary hippocampal neural stem cells were treated with stress hormone corticosterone (CORT) to reveal the potential antidepression mechanism of theacrine from the perspective of adult hippocampus neurogenesis. Results of behavioral and neurotransmitter analysis showed that intragastric administration of theacrine significantly counteracted chronic stress-induced depression-like disorders and abnormal 5-hydroxytryptamine (5-HT) metabolism with less central excitability. Further investigation from both in vivo and in vitro experiments indicated that the antidepressant mechanism of theacrine was associated with promoting adult hippocampal neurogenesis, via the modulation of the phosphodiesterase-4 (PDE4)/cyclic adenosine monophosphate (cAMP)/cAMP response-element binding (CREB)/brain-derived neurotrophic factor (BDNF)/tropomyosin-related kinase B (TrkB) pathway. Collectively, our findings could promote the prevalence of kucha as a common beverage with uses for health care and contribute to the development of theacrine as a potential novel antidepressant medicine.

Glabrone as a specific UGT1A9 probe substrate and its application in discovering the inhibitor glycycoumarin.

UDP-glucuronosyltransferase 1A9 (UGT1A9) is one of the most important UGT isoforms, and plays an important role in the metabolic elimination of therapeutic drugs via glucuronidation. Herbal medicines affecting the activity of UGT1A9 may influence the metabolism of related drugs, thus causing herb-drug interactions and even adverse effects. However, few methods are available to evaluate the activity of UGT1A9. In this study, a natural product glabrone was discovered as an isoform-specific probe substrate for UGT1A9. The Vmax and Km values of glabrone were 362.6 nmol/min/mg protein and 17.2 µM for human liver microsomes (HLMs), and 382.3 nmol/min/mg protein and 16.6 µM for recombinant human UGT1A9, respectively. Glabrone 7-O-glucuronide, the UGT1A9 metabolite of glabrone, was prepared by using a plant glucuronosyltransferase UGT88D1, and the structure was identified by NMR spectroscopy. Using glabrone as a probe, we established a rapid HPLC method to screen UGT1A9 inhibitors from 54 natural products isolated from the Chinese herbal medicine licorice. Among them, glycycoumarin was found as a potent UGT1A9 inhibitor with an IC50 value of 6.04 µM. In rats, the pretreatment of glycycoumarin (4 mg/kg, i.p.) for 3 days could remarkably increase the plasma concentrations of dapagliflozin while decrease the concentrations of dapagliflozin-O-glucuronide after administration of dapagliflozin (1 mg/kg, i.v.), which is mainly metabolized by UGT1A9. The results indicated the potential risk of herb-drug interactions between licorice and UGT1A9-metabolizing drugs.

Celastrol ameliorates Propionibacterium acnes/LPS-induced liver damage and MSU-induced gouty arthritis via inhibiting K63 deubiquitination of NLRP3.

BACKGROUND: Celastrol, a pentacyclic triterpenoid quinonemethide isolated from several spp. of Celastraceae family, exhibits anti-inflammatory activities in a variety of diseases including arthritis. PURPOSE: This study aims to investigate whether the inhibition of NLRP3 inflammasome is engaged in the anti-inflammatory activities of celastrol and delineate the underlying mechanism. METHODS: The influence of celastrol on NLRP3 inflammasome activation was firstly studied in lipopolysaccharide (LPS)-primed mouse bone marrow-derived macrophages (BMDMs) and phorbol 12-myristate 13-acetate (PMA)-primed THP-1 cells treated with nigericin. Reconstituted inflammasome was also established by co-transfecting NLRP3, ASC, pro-caspase-1 and pro-IL-1ß in HEK293T cells. The changes of inflammasome components including NLRP3, ASC, pro-caspase-1/caspase-1 and pro-IL-1ß/IL-1ß were examined by enzyme-linked immunosorbent assay (ELISA), western blotting and immunofluorescence. Furthermore, Propionibacterium acnes (P. acnes)/LPS-induced liver injury and monosodium urate (MSU)-induced gouty arthritis in mice were employed in vivo to validate the inhibitory effect of celastrol on NLRP3 inflammasome. RESULTS: Celastrol significantly suppressed the cleavage of pro-caspase-1 and pro-IL-1ß, while not affecting the protein expressions of NLRP3, ASC, pro-caspase-1 and pro-IL-1ß in THP-1 cells, BMDMs and HEK293T cells. Celastrol suppressed NLRP3 inflammasome activation and alleviated P. acnes/LPS-induced liver damage and MSU-induced gouty arthritis. Mechanism study revealed that celastrol could interdict K63 deubiquitination of NLRP3, which may concern interaction of celastrol and BRCA1/BRCA2-containing complex subunit 3 (BRCC3), and thereby prohibited the formation of NLRP3, ASC and pro-caspase-1 complex to block the generation of mature IL-1ß. CONCLUSION: Celastrol suppresses NLRP3 inflammasome activation in P. acnes/LPS-induced liver damage and MSU-induced gouty arthritis via inhibiting K63 deubiquitination of NLRP3, which presents a novel insight into inhibition of celastrol on NLRP3 inflammasome and provides more evidences for its application in the therapy of inflammation-related diseases.

Theacrine, a purine alkaloid from kucha, protects against Parkinson's disease through SIRT3 activation.

BACKGROUND: Oxidative damage of dopaminergic neurons is the fundamental causes of Parkinson's disease (PD) that has no standard cure at present. Theacrine, a purine alkaloid from Chinese tea Kucha, has been speculated to benefit the neurodegeneration in PD, through similar actions to its chemical analogue caffeine, albeit excluding side effects. Theacrine has nowadays gained a lot of interest for its multiple benefits, while the investigations are weak and insufficient. HYPOTHESIS/PURPOSE: It is well-known that tea has a wide range of functions, especially in the prevention and treatment of neurodegenerative diseases. Theacrine is an active monomer compound in Camellia assamica var. kucha Hung T. Chang & H.S.Wang (Kucha), which appears to be effective and safe in PD therapy. The aim of this study is to examine its actions in diverse PD models and explore the mechanisms. STUDY DESIGN: For determination of theacrine's effects, we employed diverse oxidative damage-associated PD models, including 6-OHDA-treated rats, MPTP-treated mice/zebrafish and MPP+-treated SH-SY5Y cells, and using caffeine, selegiline and depranyl as positve control. For investigation and verification of the mechanisms, we utilized approaches testing mitochondrial function-related parameters and enzyme activity as well as applied gene knockdown and overexpression. METHODS: We employed behavioral tests including spontaneous activity, pole, swimming, rotarod and gait, immunohistochemistry, HPLC, flow cytometry, immunohistochemistry, Western blot, gene knockdown by siRNA and overexpression by plasmid in this study. RESULTS: Theacrine is demonstrated to retrieve the loss of dopaminergic neurons and the damages of behavioral performance in multiple animal models of PD (6-OHDA-treated rats and in MPTP-treated mice and zebrafish). The followed data of MPP+-treated SH-SY5Y cells indicate that theacrine relieves apoptosis resulted from oxidative damage and mitochondrial dysfunction. Further investigations illustrate that theacrine activates SIRT3 directly. It is of advantage to prevent apoptosis through SIRT3-mediated SOD2 deacetylation that reduces ROS accumulation and restores mitochondrial function. This concept is elaborated by 3TYP that inhibits SIRT3 enzyme activity and knockdown/overexpression of SIRT3 gene, demonstrating a crucial role of SIRT3 in theacrine-benefited dopaminergic neurons. CONCLUSION: Theacrine prevents apoptosis of dopaminergic neurons through directly activating SIRT3 which deacetylating SOD2 and restoring mitochondrial functions.

Efficacy of postoperative intraperitoneal hyperthermic perfusion chemotherapy with oxaliplatin + 5-Fluorouracil in the treatment of gastric cancer patients with peritoneal carcinomatosis.

PURPOSE: To explore the effect of palliative laparoscopic resection of gastric cancer combined with intraperitoneal hyperthermic perfusion chemotherapy (IHPC) with oxaliplatin + 5-fluorouracil (5-FU) on gastric cancer patients with peritoneal carcinomatosis (PC). METHODS: 90 patients definitely diagnosed with gastric adenocarcinoma and PC and admitted to our hospital from March 2013 to March 2016 were collected and divided into IHPC group (n=45) and control group (n=45). In IHPC group, IHPC with oxaliplatin + 5-FU was carried out for the first time on the first day after operation, and then it was conducted once every other day for a total of 4 times. The clinical efficacy, quality of life, adverse reactions, postoperative tumor recurrence and survival of the patients were observed and recorded. RESULTS: The total effective rates in IHPC group and control group were 62.2% (28/45) and 55.6% (25/45), respectively (p>0.05). In both groups, the curative effect was the best in moderately differentiated adenocarcinoma and worst in signet ring cell carcinoma. Besides, the effective rates of Karnofsky performance status (KPS) in the two groups after operation were 82.2% (37/45) and 75.6% (34/45), respectively (p=0.606). However, the renal function indexes, serum creatinine (sCr) and blood urea nitrogen (BUN) in the two groups of patients after operation were increased, and those in the IHPC group were higher than those in the control group (p=0.016, p=0.010). Moreover, follow-up results of patients' survival revealed that the OS and PFS in the IHPC group were significantly higher than those in the control group (p=0.041, p=0.045). CONCLUSIONS: Palliative laparoscopic resection of gastric cancer combined with IHPC with oxaliplatin +5-FU has a definite therapeutic effect on gastric cancer with PC, which can achieve a better short-term clinical therapeutic effect and better postoperative quality of life.

Babao Dan attenuates acute ethanol-induced liver injury via Nrf2 activation and autophagy.

BACKGROUND: Babao Dan (BBD), a traditional Chinese medicine, has been used as a complementary and alternative medicine to treat multifarious liver diseases. In this study, we aimed to observe its protective effect on ethanol-induced liver injury and explore potential mechanisms. METHODS: Mice pretreated with BBD (0.125, 0.25 and 0.5 g/kg BW) were administrated by ethanol gavage (5 g/kg BW). Liver injury biomarkers and hepatic redox parameters were evaluated by histopathology as well as serum and hepatic content analysis. AML-12 cell was also utilized to determine the efficacy of BBD against ethanol-induced hepatotoxicity. RESULTS: Drunkenness experiment showed that the latency was significantly increased and the drunken sleep time was decreased in mice pretreated with BBD. We then found that BBD could reduce hepatic lipid peroxidation and steatosis induced by ethanol exposure. BBD could also suppress ethanol-induced depletion of hepatic antioxidant enzyme. Besides that, BBD treatment lessened the induction of hepatic cytochrome P450 2E1, a major contributor to ethanol-mediated oxidative stress, and up-regulated the expression of nuclear factor erythroid 2-related factor 2 and its two transcriptional targets hemeoxygenase-1 and glutamate-cysteine ligase catalytic subunit. Furthermore, autophagy induced by BBD contributed to hepatoprotection activity. CONCLUSIONS: Our results suggest that BBD can markedly dispel acute ethanol-induced hepatotoxicity through multiple pathways including attenuation of ethanol-mediated oxidative stress, enhancement of the oxidative defense systems and activation of autophagy.

Isolariciresinol-9'-O-α-L-arabinofuranoside protects against hydrogen peroxide­induced apoptosis of human umbilical vein endothelial cells via a PI3K/Akt/Bad­dependent pathway.

Isolariciresinol-9'-O-α-L-arabinofuranoside (MWS­19) isolated from Pinus massoniana Lamb. Fresh pine needles is the major ingredient of the Songling Xuemaikang capsule therapy used for hypertension. The present study aimed to investigate the effects and underlying mechanisms of MWS­19 on hydrogen peroxide (H2O2)­induced apoptosis in human umbilical vein endothelial cells (HUVECs). To investigate the effect of MWS­19 on apoptosis in HUVECs, an oxidative stress­induced apoptosis model was established in HUVECs using H2O2, and the present study performed Hoechst 33258 staining and a Cell Counting Kit­8 (CCK­8) assay. Furthermore, western blot analysis was also performed to investigate the underlying mechanism of the effects of MWS­19 on the model. The results demonstrated that MWS­19 reversed the effects of H2O2 on cell apoptosis at a concentration range of 15.6­250 µg/ml, with dose­dependent increases in cell growth. Hoechst staining indicated that 500 µM H2O2 induced HUVEC apoptosis, and MWS­19 markedly protected HUVECs against apoptosis at 31.3, 62.5 and 125 µg/ml. Furthermore, the protein expression of phosphatidylinositol 3­kinase (PI3K), phosphorylated­Akt and Bcl­2­associated agonist of cell death (Bad) were increased, and reduced caspase­3 activation was observed, following treatment with MWS­19 in H2O2­treated HUVECs. Additionally, the PI3K inhibitor wortmannin attenuated PI3K/Akt/Bad signaling induced by MWS­19 treatment and neutralized the effect of MWS­19 on the growth of HUVECs. In conclusion, the results of the present study indicate that MWS­19 may protect against H2O2­induced HUVEC apoptosis via the PI3K/Akt/Bad signaling pathway. MWS­19 may serve an important role in the prevention of oxidative damage in vascular endothelial cells in hypertension patients.

Protective effects of Oviductus Ranae-containing serum on oxidative stress-induced apoptosis in rat ovarian granulosa cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Oviductus Ranae (OR) is a traditional Chinese medicine derived from Rana temporaria chensinensis David, and is known to have a wide variety of pharmacological effects. AIM OF THE STUDY: To investigate the function and mechanism of OR-containing serum in protecting rat ovarian granulosa cells from hydrogen peroxide (H2O2)-induced oxidative damage. MATERIALS AND METHODS: H2O2-treated granulosa cells were pretreated with OR-containing serum, and viability and proliferation assays were carried out using Cell Counting Kit-8 (CCK-8). Apoptotic granulosa cells were observed microscopically using 4',6-diamidino-2-phenylindole (DAPI), and the apoptotic ratio was quantified via Annexin V/ propidium iodide (PI) staining combined with flow cytometry. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) in the cells were measured using 2,7-dichlorofluorescein diacetate (DCFH-DA) and rhodamine 123, respectively, and analyzed by flow cytometry. Mitogen-activated protein kinases (MAPKs), including ERK1/2, JNK, and p38, and other apoptosis-related proteins (p53, Bcl-2, Bax, caspase-9, caspase-3), were detected by western blot analysis, and the related mRNA levels were detected using reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The results revealed that treatment with OR-containing serum reduced apoptosis and mitochondrial membrane damage in H2O2-treated granulosa cells. The OR-containing serum interfered with H2O2-induced intracellular generation of ROS and loss of ΔΨm, which typically lead to apoptosis. Furthermore, the OR-containing serum down-regulated pro-apoptotic proteins such as p53, Bax, caspase-9, and caspase-3, while up-regulating the anti-apoptotic protein Bcl-2. Finally, the OR-containing serum increased phosphorylation of ERK1/2, and reduced JNK and p38 phosphorylation. CONCLUSIONS: OR-containing serum protected rat ovarian granulosa cells against H2O2-induced apoptosis, by reducing ROS production and improving mitochondrial membrane potential, through down-regulation of negative regulators of proliferation, activation of ERK1/2, and inhibition of the activity of JNK and p38.

Discovery of a cobalt complex with high MEK1 binding affinity.

A series of Schiff base ligands (L1-L5) and their cobalt(II) complexes (1-5) were designed and synthesized for MEK1 binding experiment. The biological evaluation results showed that Bis(N,N'-disalicylidene)-3,4-phenylenediamine-cobalt(II) 1 and Bis(N,N'-disalicylidene)-1,2-cyclohexanediamine-cobalt(II) 2 are much more effective than the parent Schiff bases (L1 and L2). Importantly, 2 exhibited MEK1 binding affinity with IC5071nM, which is so far the best result for metal complexes and more potent than U0126 (7.02µM) and AZD6244 (2.20µM). Docking study was used to elucidate the binding modes of complex 2 with MEK1. Thus cobalt(II) complex 2 may be further developed as a novel MEK1 inhibitor.

Babao Dan attenuates hepatic fibrosis by inhibiting hepatic stellate cells activation and proliferation via TLR4 signaling pathway.

Babao Dan (BBD), a traditional Chinese medicine, has been widely used as a complementary and alternative medicine to treat chronic liver diseases. In this study, we aimed to observe the protective effect of BBD on rat hepatic fibrosis induced by diethylnitrosamine (DEN) and explore it possible mechanism. BBD was administrated while DEN was given. After eight weeks, values of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) indicated that BBD significantly protected liver from damaging by DEN and had no obvious side effect on normal rat livers. Meanwhile, BBD attenuated hepatic inflammation and fibrosis in DEN-induced rat livers through histopathological examination and hepatic hydroxyproline content. Furthermore, we found that BBD inhibited hepatic stellate cells activation and proliferation without altering the concentration of lipopolysaccharide (LPS) in portal vein. In vitro study, serum from BBD treated rats (BBD-serum) could also significantly suppress LPS-induced HSCs activation through TLR4/NF-κB pathway. In addition, BBD-serum also inhibited the proliferation of HSCs by regulating TLR4/ERK pathway. Our study demonstrated that BBD may provide a new therapy strategy of hepatic injury and hepatic fibrosis.