Telocinobufagin, a PLK1 suppressor that inhibits tumor growth and metastasis by modulating CDC25c and CTCF in HNSCC cells.

BACKGROUND: The high metastasis and mortality rates of head and neck squamous cell carcinoma (HNSCC) urgently require new treatment targets and drugs. A steroidal component of ChanSu, telocinobufagin (TBG), was verified to have anti-cancer effects in various tumors, but its activity and mechanism in anti-HNSCC were still unknown. PURPOSE: This study tried to demonstrate the anti-tumor effect of TBG on HNSCC and verify its potential mechanism. METHODS: The effect of TBG on cell proliferation and metastasis were performed and the TBG changed genes were detected by RNA-seq analysis in HNSCC cells. The GSEA and PPI analysis were used to identify the pathways targeted for TBG-regulated genes. Meanwhile, the mechanism of TBG on anti-proliferative and anti-metastasis were investigated in vitro and in vivo. RESULTS: The in vitro and in vivo experiments confirmed that TBG has favorable anti-tumor effects by induced G2/M phase arrest and suppressed metastasis in HNSCC cells. Further RNA-seq analysis demonstrated the genes regulated by TBG were enriched at the G2/M checkpoint and PLK1 signaling pathway. Then, the bioinformatic analysis of clinical data found that high expressed PLK1 were closely associated with poor overall survival in HNSCC patients. Furthermore, PLK1 directly and indirectly modulated G2/M phase and metastasis (by regulated CTCF) in HNSCC cells, simultaneously. TBG significantly inhibited the protein levels of PLK1 in both phosphorylated and non-phosphorylated forms and then, in one way, inactivated PLK1 failed to activate G2/M phase-related proteins (including CDK1, CDC25c, and cyclin B1). In another way, be inhibited PLK1 unable promote the nuclear translocation of CTCF and thus suppressed HNSC cell metastasis. In contrast, the anti-proliferative and anti-metastasis effects of TBG on HNSCC cell were vanished when cells high-expressed PLK1. CONCLUSION: The present study verified that PLK1 mediated TBG induced anti-tumor effect by modulated G2/M phase and metastasis in HNSCC cells.

Sanggenon C Suppresses Tumorigenesis of Gastric Cancer by Blocking ERK-Drp1-Mediated Mitochondrial Fission.

Sanggenon C is a flavonoid extracted from the root bark of white mulberry, which is a traditional Chinese medicine with anti-inflammatory, antioxidative, and antitumor pharmacological effects. In this study, sanggenon C was found to inhibit human gastric cancer (GC) cell proliferation and colony formation, induce GC cell cycle arrest in the G0-G1 phase, and promote GC cell apoptosis. Moreover, sanggenon C was found to decrease the level of mitochondrial membrane potential in GC cells and inhibit mitochondrial fission. Mechanistically, RNA sequencing, bioinformatics analysis, and a series of functional analyses confirmed that sanggenon C inhibited mitochondrial fission to induce apoptosis by blocking the extracellular regulated protein kinases (ERK) signaling pathway, and constitutive activation of ERK significantly abrogated these effects. Finally, sanggenon C was found to suppress the growth of tumor xenografts in nude mice without obvious side effects to the vital organs of animals. This study reveals that sanggenon C could be a novel therapeutic strategy for GC treatment.

Characterization of the complete chloroplast genome of Euphorbia pekinensis Rupr. (Euphorbiaceae).

Euphorbia pekinensis Rupr. 1859 is a medicinal herb endemic to China and distributed throughout the country, particularly across the northern part of the mainland. However, the systematic classification of Euphorbiaceae remains controversial. Therefore, studying the chloroplast genome of E. pekinensis is crucial for the resolution of this taxonomic dispute, clarification of the systematic status of Euphorbia, and establishment of an accurate classification system for Euphorbiaceae. In this study, we sequenced the complete chloroplast genome of E. pekinensis using Illumina sequencing technology and annotated it using GeSeq. The complete chloroplast genome was 162,002-bp-long with a guanine-cytosine (GC) content of 35.7%. It included one large single-copy (LSC), one small single-copy (SSC), and two inverted repeat sequence regions (IRa and IRb), which were 90,225 bp, 18,067 bp, and 26,855 bp in length, respectively, and are indicative of a typical tetrad structure. The genome encoded 129 functional genes, comprising 85 protein-coding genes, 36 tRNA genes, and eight rRNA genes. According to the maximum-likelihood phylogenetic tree that was constructed using 16 complete chloroplast genomes, E. pekinensis was found to be closely related to E. ebracteolata. Therefore, the complete chloroplast genome of E. pekinensis provides a better understanding of Euphorbia genetics.

Study on Network Pharmacological Analysis and Preliminary Validation to Understand the Mechanisms of Plantaginis Semen in Treatment of Gouty Nephropathy.

Plantaginis Semen (PS) has been used to promote diuresis and clear away dampness. Recent reports have shown that PS can be used to treat gouty nephropathy (GN). However, the action and mechanism of PS have not been well defined in treating GN. The present study aimed to define the molecular mechanisms of PS as a potential therapeutic approach to treat GN. A combination of network pharmacology and validation experiments in GN is used to understand the potential mechanism. Information on pharmaceutically active compounds in PS and gene information related to GN was obtained from public databases. The compound target network and protein-protein interaction network were constructed to study the mechanism of action of PS in the treatment of GN. The mechanism of action of PS in the treatment of GN was analyzed via Gene Ontology (GO) biological process annotation and Kyoto Gene and Genomics Encyclopedia (KEGG) pathway enrichment. Validation experiments were performed to verify the core targets. The GN rat model was prepared by the method of combining yeast and adenine. Hematoxylin-eosin (HE) staining was used to observe the morphology of renal tissue in rats. ELISA was applied to detect TGF-ß1, TNF-α, and IL-1ß levels in renal tissue. The expressions of TGF-ß1, TNF-α, and IL-1ß were determined using immunohistochemistry. Through the results of network pharmacology, we obtained 9 active components, 118 predicted targets, and 149 GN targets from the public database. Based on the protein-protein interaction (PPI), 26 hub genes for interaction with PS treating for GN were screened, including MMP9, TNF, IL1ß, and IL6. The enrichment analysis results showed that the treatment of GN with PS was mainly involved in the TGF-ß1 signaling pathway, MAPK signaling pathway, TNF signaling pathway, NF-κB signaling pathway, and PI3K Akt signaling pathway. Validation experiment results showed that PS could reduce the content of urinary protein and UA and deregulate the expression of TGF-ß1, TNF-α, and IL-1ß in the treatment of GN. The molecular mechanism of PS in the treatment of GN indicated the synergistic features of multicomponent, multitarget, and multipathway of traditional Chinese medicine, which provided an essential scientific basis for further elucidating the mechanism of PS in the treatment of GN.

Influence of artificial luminous environment and TCM intervention on development of myopia rabbits.

OBJECTIVE: To explore the influence of artificial luminous environment and preventive function of traditional Chinese medicine (TCM) intervention based on "Theory of yin-yang clock" on myopia. METHODS: A total of 45 New Zealand young rabbits were randomly divided into 5 groups, 9 for each group. Control group was exposed in natural light. Fluorescent group and full spectrum group were exposed in fluorescent light and full spectrum light, on which basis fluorescent TCM group and full spectrum TCM group were added with "Rizhong Yinyang Formulas", respectively. Optical parameters were measured and the influence of different lights on the serum and retinal dopamine (DA) levels as well as the retinal histopathological tissues was observed. RESULTS: The spectrum of fluorescent light mainly focused at 420-490 nm with the peak value of wavelength near 450 nm, whereas that of full spectrum was wider (400-800 nm) with the peak value near 600 nm. After 4 and 12 weeks, fluorescent group was evidently lower in serum and retinal DA levels (P<0.01), and there was no significant difference among full spectrum group, fluorescent TCM group and full spectrum TCM group (P>0.05). Histopathological observation showed that there was significant difference in pigment epithelium layer, photoreceptor and nerve fiber layer between fluorescent group and control group, but the difference among the test groups was not significant. CONCLUSIONS: Fluorescent light has certain influence on retinal histological construction and visual performance. However, TCM intervention may have some degree of protective function on retina.

Anti-inflammatory and analgesic activity of total flavone of Cunninghamia lanceolata.

The present study was undertaken to investigate the anti-inflammatory and analgesic activity of total flavone of branches and leaves of Cunninghamia lanceolata (TFC) to provide a scientific basis for its clinical use and resource development. TFC was evaluated for anti-inflammatory and analgesic activity in mice or rats using chemical and thermal models of nociception, including acetic acid-induced writhing test, hot plate latency test, formalin test and carrageenan induced paw oedema test. Results showed that TFC given orally can significantly attenuate acetic acid-induced writhing in mice in a dose-dependent manner. In the hot plate latency test, TFC showed common activity in prolonging duration time only at the highest dose (400 mg/kg). Each dose of TFC could not significantly inhibit the first phase but was active in the later phase of formalin-induced pain, whereas morphine showed notable activity in the two phases. In the carrageenan-induced paw oedema model, TFC could significantly and dose-dependently reduce the carrageenan-induced paw edema at the third and fifth hour, and decrease the content of PEG(2) in paw edema tissue and that of COX-2 in blood serum. It may be concluded that TFC showed both anti-inflammatory and analgesic effects, showing that it can be of importance in drug development, especially in the field of pain and inflammation.

Identification of a novel function of Id-1 in mediating the anticancer responses of SAMC, a water-soluble garlic derivative, in human bladder cancer cells.

Studies have shown that the expression of inhibitor of differentiation (Id-1) is increased in bladder cancer and is associated with drug resistance. S-allylmercaptocysteine (SAMC), a water-soluble component of garlic, is known to have a potent therapeutic effect on human cancer. The aim of this study was to investigate whether Id-1 expression mediates SAMC-induced cell death in bladder cancer cells. After generating stable Id-1-expressing and si-Id-1 transfectants in various bladder cancer cell lines, cell sensitivity to SAMC was compared by colony formation and MTT assays. The results indicated that Id-1 overexpression reduced the positive effect of SAMC on cell survival, while the inactivation of Id-1 increased cellular susceptibility to SAMC. Using DAPI staining, the apoptosis of bladder cancer cells induced by SAMC was shown to be negatively regulated by Id-1 expression. The expression of apoptosis-related proteins analyzed by Western blotting further supported the negative role of Id-1 in SAMC-induced apoptosis. Furthermore, by wound closure and type I collagen invasion assays, the inhibitory effect of SAMC on the invasion and migration of bladder cancer cells was found to be associated with the down-regulation of Id-1. Our results demonstrated that SAMC-induced apoptosis is associated with the Id-1 pathway, and that the inactivation of Id-1 enhances the ability of SAMC to inhibit the survival, invasion and migration of bladder cancer cells. These findings may lead to the development of novel therapeutic strategies for the treatment of bladder cancer.

Effect of qiming granule on retinal blood circulation of diabetic retinopathy: a multicenter clinical trial.

OBJECTIVE: To objectively assess the effect of Qiming Granule in the treatment of diabetic retinopathy (DR) by fluorescence fundus angiography (FFA). METHODS: In a multi-center, randomized, parallel controlled clinical trial, patients with DR were randomly assigned to the control group (calcium dobesilate capsule) and the test group (Qiming Granule). Changes in the retinal blood circulation time were recorded by FFA after 3 months of medication. RESULTS: Significant reduction was observed in the retinal arterio-venous circulation time (AVCT) in both groups (P<0.01), the value was 7.635+/-3.149 s before treatment and 5.165 +/-3.382 s after treatment in the treated group, and 7.737+/-3.413 s and 5.313+/-3.472 s in the control group respectively. Qiming Granule also reduced the arm-to-retinal circulation time (ARCT, P<0.05). The value was 17.867+/-3.872 s before treatment and 15.643+/-4.648 s after treatment in the treated group, and 17.217+/-3.833 s and 16.312+/-3.613 s in the control group (P>0.05) respectively. The ARCT in the tested group was reduced, with a statistically significant difference post-medication (P<0.01). CONCLUSION: As a Chinese medicine complex prescription, Qiming Granule may alleviate retinal hypoxia and ischemia by increasing retinal blood flow and improving the blood circulation.

Bakkenolides from Petasites tricholobus and their neuroprotective effects related to antioxidant activities.

Four novel bakkenolides - bakkenolide-Ia ( 1), bakkenolide-IIa ( 2), bakkenolide-IIIa ( 3) and bakkenolide-IVa ( 4) - were isolated from the extract of the rhizome of Petasites tricholobus. The structures were characterized by using NMR ( (1)H, (13)C, (1)H- (1)H COSY, HMQC, HMBC, and NOESY) and mass spectrometry. The neuroprotective activity of the compounds 1 - 4 was assayed with primary cultured neurons exposed to oxygen-glucose deprivation and oxidative insults. Antioxidant activity of the bakkenolides was evaluated by cell-free bioassays. The IN VITRO assay results showed that all these compounds exhibited significant neuroprotective and antioxidant activities. To our knowledge, this is the first report on the neuroprotective and antioxidant activities of bakkenolides.

[Effects of EGb761 on hippocamal synaptic plasticity of vascular dementia rats].

AIM: To investigate the effects of ginkgo biloba extract 761 (EGb761) on synaptic plasticity in hippocampus of vascular dementia (VD) rats. METHODS: The escape latency (EL) of Morris water maze (MWM) task was measured at different time points (4 W, 8 W and 16 W), and the population spikes (PS) of granule cell layer in the dentate gyrus were induced by single pulse stimulation to perfo rate path fibers before and after high frequency stimulation (HFS) in vivo. RESULTS: MWM test showed that the escape latency (EL) of VD model group were highly longer than that of the sham-operated group, while the EL of EGb761-treated group was significantly shorter than that of model group, but still longer than that of the sham-operated group. The incidence rates of LTP induction in 1 m, 2 m and 4 m subgroups of model group were significantly lower than that of sham-operated group and EGb761-treated group at different time point. The relative amplitudes of PS after HFS in 1 m, 2 m and 4 m subgroups of model group were obviously reduced compared with that of the corresponding subgroups of sham-operated group and EGb761-treated group. There was no obvious significance in the peak latency of PS between different subgroups and different LTP-tested time point. CONCLUSION: VD model rats had apparent and long-lasting dysfunction of learning and memory, EGb761 could accelerate the recovery of the pathological synaptic plasticity. This suggested that EGb761 played an important and improving role on learning and memory dysfunction of VD.

[Initial discussion of mice acute hepatic injury caused by Tripterygium glycosides].

OBJECTIVE: To discuss the mechanism of mice acute hepatic injury caused by tripterygium glycosides tablets with different dose at different time. METHOD: Mice were given Tripterygium Glycosides respectively with the dosage of 10 times, 20 times, 30 times of clinical dose to observe the change of mice acute hepatic injury with different does; then, the acute hepatic injury mice were duplicated with 20 times clinical dosage and mice serum ALT were detected at 9, 18, 27, 36 h to observe the change of mice acute hepatic injury at different time. The activity of SOD, GSH-Px in serum and the level of LPO in liver homogenate wevedetected to discuss the mechanism of mice acute hepatic injury caused by Tripterygium Glycosides tablets; and liver tissue pathology was observed. RESULT: The acute hepatic injury was obvious with 20 times adult dosage in 18 hours and the acute hepatic injury mice death rate was low. CONCLUSION: Tripterygium Glycosides tablets can cause acute hepatic injury to mice and its mechanism is related to Lipid peroxidation reaction.