Investigating the pharmacological mechanism of Zhengyuan jiaonang for treating colorectal cancer via network pharmacology analysis and experimental verification.

ETHNOPHARMACOLOGICAL RELEVANCE: Zhengyuan jiaonang (ZYJN) is a traditional Chinese patent medicine (CPM) used in China for adjuvant cancer therapy, which has been proved to have anti-fatigue effects. AIM OF STUDY: The study aims to investigate the antitumor effects of ZYJN and its underlying mechanisms using subcutaneous transplant CT26 model. MATERIALS AND METHODS: Fingerprint analysis of ZYJN was performed using high performance liquid chromatography. The potential targets of ZYJN were predicted using bioinformatic analysis, which were further validated by Western Blot assay. Subcutaneous transplant CT26 model was used to evaluate the antitumor effects of ZYJN. The effects of ZYJN on the tumor immune microenvironment were investigated by flow cytometry. Transparent imaging was used to investigate the effects of ZYJN on fibrosis and angiogenesis. RESULTS: ZYJN could inhibit colorectal cancer growth when administered alone or in combination with 5-FU. The combination of ZYJN and 5-FU could significantly increase the serum level of albumin (ALB) and decrease the serum level of aspartate aminotransferase (AST). In addition, the combination of ZYJN at 0.75 g/kg and 5-FU significantly decreased the serum level of vascular endothelial growth factors (VEGF) and inhibited the angiogenesis of CT26 cancer. The combination of ZYJN at 1.50 g/kg and 5-FU could promote the fibrosis process of CT26 cancer. Additionally, combination of ZYJN and 5-FU could significantly increase the percentage of tumor-infiltrating T cells and CD4+ T cells in the late stage of CT26 model, while ZYJN at 1.50 g/kg increased the percentage of NK cells as well as CD8+ T cells in the early stage of CT26 model. Western Blot analysis revealed that administration of ZYJN at 0.75 g/kg reduced the expression of PI3K-p110α, CDK1, CCNB1 and MMP-9, and inhibited the phosphorylation of Akt (Thr308). CONCLUSIONS: ZYJN could inhibit the tumor growth of CT26 colorectal cancer by promoting tumor fibrosis, suppressing angiogenesis, migration, and invasion and modulating the tumor immune microenvironment. ZYJN enhanced the efficacy and reduced the toxicity of chemotherapy drugs in combination therapy. Our findings provide evidence for the clinical application of ZYJN in cancer treatment.

Effects of light and water disturbance on the growth of Microcystis aeruginosa and the release of algal toxins.

Eutrophication of water constitutes a serious threat to global water quality. Light intensity and water disturbance are important factors affecting the growth of algae and the release of algal toxins. In this study, algal growth indicators, algal enzyme systems, and algal toxin release in Microcystis aeruginosa under different light intensities and water disturbances were determined. The results showed that 2500 lx and 120 rpm were the optimal conditions for the growth of M. aeruginosa. The growth of algal cells was inhibited by high light intensity and high water disturbance. However, the optimal conditions for algal growth were not favorable conditions for the release of algal toxin. The highest concentration of microcystin-LR (MC-LR), observed at 4500 lx and 80 rpm, was 198.1 µg/L, whereas the highest single cell toxin production reached up to 10.49 × 10-9  µg/cell at 7000 lx and 120 rpm. Redundancy analysis results showed that the concentration of MC-LR was positively correlated with algal cell density and antioxidant enzyme activities (superoxide dismutase, catalase, peroxidase, and malondialdehyde [MDA]) and negatively correlated with the total nitrogen and total phosphorus consumption rates and MDA. Single cell toxin production was negatively correlated with algal cell density and antioxidant enzyme activity but positively correlated with MDA content. PRACTITIONER POINTS: There was an optimal water disturbance condition for algae growth affected by the light intensity. Optimal conditions for algae cell growth are not necessarily the optimal conditions for algal toxin release. Enzyme indicators have correlation with the release of algae toxins and the growth of algae cells.

Toxicokinetics and toxicity of potassium 2-(1-hydroxypentyl)-benzoate in beagle dogs.

Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB) is a prodrug of 3-n-butylphthalide (dl-NBP) for treatment of cerebral ischemic stroke in China, which undergoes lactonization to form dl-NBP in plasma. And, the phase II-III clinical trial of dl-PHPB has been approved by China Food and Drug Administration (CFDA) in 2013. In this study, a toxicity and toxicokinetics evaluation of dl-PHPB was performed using beagle dogs at specially high-dose 108 mg/kg/day (65-fold higher than humans at MHRD) for 4 weeks by intravenous administration, with a 3-week recovery period. And the plasma concentrations of dl-PHPB along with its metabolite dl-NBP were determined by HPLC-UV method. The results showed that dl-PHPB was quickly metabolized into dl-NBP, and no significant accumulation was observed. A slight to moderate behavior-associated toxicity was revealed in the process of delivery; and recovered to normal at the end of administration. Changes in the blood hematological profiles included significantly increased NEUT levels and lower LYM% content. Meanwhile, a notable increase in TG content was also observed in the serum biochemical parameters at 4-week post-exposure. These findings were reversible during the recovery period. The information from these studies would be taken into consideration for the interpretation of toxicology findings and provide a reference for clinical safety assessment.

Can Aidi injection alleviate the toxicity and improve the clinical efficacy of radiotherapy in lung cancer?: A meta-analysis of 16 randomized controlled trials following the PRISMA guidelines.

BACKGROUND/INTRODUCTION: Aidi injection plus radiotherapy is widely used for lung cancer in China. Can Aidi injection alleviate the toxicity and improve the clinical efficacy of radiotherapy in lung cancer? Has Aidi injection the attenuation and synergistic efficacy to radiotherapy? There is lack of strong evidence to prove it. OBJECTIVES: To reveal its real attenuation and synergistic efficacy to radiotherapy and provide sufficient evidence for adjuvant chemotherapy strategies to lung cancer, we systematically evaluated all related studies. DATA SOURCES: We collected all studies about Aidi injection plus radiotherapy for lung cancer in Medline, Embase, Web of Science, China national knowledge infrastructure database (CNKI), Chinese scientific journals full-text database (VIP), Wanfang database, China biological medicine database (CBM) (established to June 2015), and Cochrane Central Register of Controlled Trials (June 2015), evaluated their quality according to the Cochrane evaluation handbook of randomized controlled trials (5.1.0), extracted data following the PICO principles and synthesized the data by Meta analysis. RESULTS: Sixteen randomized controlled trials (RCTs) with 1192 lung cancer patients were included, with general methodological quality in most trials. The merged relative risk (RR) values and their 95% CI of meta-analysis for objective response rate (ORR), disease control rate (DCR), and quality of life (QOL) were as follows: 1.54, (1.39,1.70), 1.10 (1.02, 1.19), and 2.13 (1.68, 2.68). The merged RR values and their 95% CI of meta-analysis for myelosuppression and neutropenia, radiation pneumonitis, and radiation esophagitis were as follows: 0.51 (0.38, 0.69), 0.53 (0.42, 0.65), 0.52 (0.41, 0.67), and 0.52 (0.40, 0.68). All were statistically significant. The possibility of publication bias was small which objectively reported the results. CONCLUSIONS: The evidence available indicates that Aidi injection plus radiotherapy can significantly improve the clinical efficacy and QOL of patients with lung cancer. Aidi injection can alleviate the myelosuppression, radiation pneumonitis, and radiation esophagitis of radiotherapy. It has the attenuation and synergistic efficacy to radiotherapy.

L-3-n-butylphthalide Rescues Hippocampal Synaptic Failure and Attenuates Neuropathology in Aged APP/PS1 Mouse Model of Alzheimer's Disease.

AIMS: Our previous studies showed that L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), improved cognitive ability in animal models of cerebral ischemia, vascular dementia, and Alzheimer's disease (AD). It is well known that cognitive deficit of AD is caused by synaptic dysfunction. In this study, we investigated the effect of L-NBP on hippocampal synaptic function in APP/PS1 AD transgenic mice and related mechanisms. METHODS: Eighteen-month-old APP/PS1 transgenic (Tg) mice were administrated 15 mg/kg L-NBP by oral gavage for 3 months. Synaptic morphology and the thickness of postsynaptic density (PSD) in hippocampal neurons were investigated by electron microscope. The dendritic spines, Aß plaques, and glial activation were detected by staining. The expressions of synapse-related proteins were observed by Western blotting. RESULTS: L-NBP treatment significantly increased the number of synapses and apical dendritic thorns and the thickness of PSD, increased the expression levels of synapse-associated proteins including PSD95, synaptophysin (SYN), ß-catenin, and GSK-3ß, and attenuated Aß plaques and neuroinflammatory responses in aged APP/PS1 Tg mice. CONCLUSION: L-NBP may restore synaptic and spine function in aged APP Tg mice through inhibiting Aß plaques deposition and neuroinflammatory response. Wnt/ß-catenin signaling pathway may be involved in L-NBP-related restoration of synaptic function.

[Structure-activity relationship of gastrodin and parishins on learning and memory deficits induced by scopolamine].

Gastrodin, parishin and parishin C were purified from a water extract of GE (rhizome of Gastrodia elata, an herb medicine for treatment of neuronal disorders). In order to compare the pharmacological effects of gastrodin, parishin and parishin C on improving cognition deficits, we tested them in an animal model of cognition disorders induced by scopolamine and in a study of in vivo long-term potentiation (LTP) recordings. In the Morris water maze task, parishin C (15 and 50 mg·kg(-1), P<0.05) and parishin (150 mg·kg(-1), P<0.05), improved spatial learning and memory significantly. However, gastrodin showed no significant effects at the dose of 150 mg·kg(-1). In vivo LTP recordings showed that parishin C at 5,10 and 20 mg·kg(-1), parishin at 10, 30 and 100 mg·kg(-1) reversed the suppression of LTP by scopolamine in rats in a dose-dependent manner. However, gastrodin at 100 mg·kg(-1) showed only a modest effect. In summary, the action of parishin C in the improvement of dementia induced by scopolamine was more potent than parishin and gastrodin.

L-3-n-butylphthalide reduces tau phosphorylation and improves cognitive deficits in AßPP/PS1-Alzheimer's transgenic mice.

L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), has been shown to have neuroprotective effects on cerebral ischemic, vascular dementia and amyloid-ß (Aß)-induced animal models by inhibiting oxidative injury, neuronal apoptosis and glial activation, regulating amyloid-ß protein precursor (AßPP) processing and reducing Aß generation. The aim of the present study was to examine the effect of L-NBP on learning and memory in AßPP and presenilin 1 (PS1) double-transgenic AD mouse model (AßPP/PS1) and the mechanisms of L-NBP in reducing Aß accumulation and tau phosphorylation. Twelve-month old AßPP/PS1 mice were given 15 mg/kg L-NBP by oral gavage for 3 months. L-NBP treatment significantly improved the spatial learning and memory deficits compared to the vehicle-treated AßPP/PS1 mice, whereas L-NBP treatment had no effect on cerebral Aß plaque deposition and Aß levels in brain homogenates. However, we found an L-NBP-induced reduction of tau hyperphosphorylation at Ser199, Thr205, Ser396, and Ser404 sites in AßPP/PS1 mice. Additionally, the expressions of cyclin-dependent kinase and glycogen synthase kinase 3ß, the most important kinases involved in tau phosphorylation, were markedly decreased by L-NBP treatment. The effects of L-NBP on decreasing tau phosphorylation and kinases activations were further confirmed in neuroblastoma SK-N-SH cells overexpressing wild-type human AßPP695 (SK-N-SH AßPPwt). L-NBP shows promising candidate of multi-target neuronal protective agent for the treatment of Alzheimer's disease.

L-3-n-butylphthalide regulates amyloid precursor protein processing by PKC and MAPK pathways in SK-N-SH cells over-expressing wild type human APP695.

Amyloid precursor protein (APP) is cleaved by α-secretase, within the amyloid-ß (Aß) sequence, resulting in the release of a secreted fragment (αAPPs) and precluding Aß production. We investigated the effects of a promising anti-AD new drug, l-3-n-butylphthalide (L-NBP), on APP processing and Aß generation in neuroblastoma SK-N-SH cells overexpressing wild-type human APP695. L-NBP significantly increased αAPPs release, and reduced Aß generation. The steady-state full-length APP levels were unaffected by L-NBP. It suggested that L-NBP regulated APP processing towards to the non-amyloidogenic α-secretase pathway. Protein kinase C (PKC) and mitogen activated protein (MAP) kinase might be involved in L-NBP-induced αAPPs secretion. L-NBP significantly increased PKCα and ɛ activations, lowered PKCγ activation and increased the phosphorylation of p44/p42 MAPK. Furthermore, PKC and MAPK inhibitors partially reduced L-NBP-induced αAPPs secretion. The results suggested alternative pharmacological mechanisms of L-NBP regarding the treatment of Alzheimer's disease (AD).

[Chemopreventive effect of resveratrol to cancer].

BACKGROUND & OBJECTIVE: Carcinogenesis is a complex process and at least 3 stages, including initiation, promotion, and progression, have been proposed in the process of carcinogenesis. Resveratrol has attracted considerable attention due to its low toxicity and unique chemical structure. This study was designed to test chemopreventive effect of resveratrol to cancer using various animal models. METHODS: Ames assay and micronucleus formation assay were used to test the antimutagenic activities of resveratrol. Croton oil-induced enhancement of ornithine decarboxylase (ODC) activities of dorsal epidermis cells in mouse and mouse ear edema model were used to investigate the anti-promotion effect of resveratrol. In addition,7,12-dimenthylbenz[a]anthracene (DMBA)/croton oil-induced mouse skin tumor model was used to evaluate chemopreventive effect of resveratrol to cancer in vivo. RESULTS: In Ames test,100 microg/plate of resveratrol exhibited 42.2% of inhibition on the reversion of Salmonella typhimurium TA100 induced by methylmethansulfonate, and 200 microg/plate of resveratrol exhibited 91.8% of inhibition on the reversion induced by benzopyrene. Pretreatment of resveratrol prevented cyclophosphamide (CTX)-induced micronucleus formation of polychromatic erythrocytes of mice bone marrow in dose-dependent manner. Mice treated with 30 mg/kg of resveratrol for 6 days before croton oil exposure have palliative ear edema. Treatment of 180 mg/kg resveratrol for 3 days caused 69.3% decrease of ODC activities in croton oil-induced dorsal epidermis. It was shown that resveratrol could inhibit DMBA/croton oil-induced mouse skin papilloma, which includes prolonging the latent period of tumor occurrence, decreasing the incidence of papilloma, and reducing tumor number per mouse in dose-dependent manner. CONCLUSION: Resveratrol has the ability of anti-mutation and anti-promotion of cancer and merit further studies as a potential cancer chemopreventive agent.