Transcriptome and metabolome analyses reveal chlorogenic acid accumulation in pigmented potatoes at different altitudes.

Pigmented potato tubers are abundant in chlorogenic acids (CGAs), a metabolite with pharmacological activity. This article comprehensively analyzed the transcriptome and metabolome of pigmented potato Huaxingyangyu and Jianchuanhong at four altitudes of 1800 m, 2300 m, 2800 m, and 3300 m. A total of 20 CGAs and intermediate CGA compounds were identified, including 3-o-caffeoylquinic acid, 4-o-caffeoylquinic acid, and 5-o-caffeoylquinic acid. CGA contents in Huaxinyangyu and Jianchuanhong reached its maximum at an altitude of 2800 m and slightly decreased at 3300 m. 48 candidate genes related to the biosynthesis pathway of CGAs were screened through transcriptome analysis. Weighted gene co-expression network analysis (WGCNA) identified that the structural genes of phenylalanine deaminase (PAL), coumarate-3 hydroxylase (C3H), cinnamic acid 4-hydroxylase (C4H) and the transcription factors of MYB and bHLH co-regulate CGA biosynthesis. The results of this study provide valuable information to reveal the changes in CGA components in pigmented potato at different altitudes.

Screening and functional analysis of StMYB transcription factors in pigmented potato under low-temperature treatment.

MYB transcription factors play an extremely important regulatory role in plant responses to stress and anthocyanin synthesis. Cloning of potato StMYB-related genes can provide a theoretical basis for the genetic improvement of pigmented potatoes. In this study, two MYB transcription factors, StMYB113 and StMYB308, possibly related to anthocyanin synthesis, were screened under low-temperature conditions based on the low-temperature-responsive potato StMYB genes family analysis obtained by transcriptome sequencing. By analyzed the protein properties and promoters of StMYB113 and StMYB308 and their relative expression levels at different low-temperature treatment periods, it is speculated that StMYB113 and StMYB308 can be expressed in response to low temperature and can promote anthocyanin synthesis. The overexpression vectors of StMYB113 and StMYB308 were constructed for transient transformation tobacco. Color changes were observed, and the expression levels of the structural genes of tobacco anthocyanin synthesis were determined. The results showed that StMYB113 lacking the complete MYB domain could not promote the accumulation of tobacco anthocyanins, while StMYB308 could significantly promote the accumulation involved in tobacco anthocyanins. This study provides a theoretical reference for further study of the mechanism of StMYB113 and StMYB308 transcription factors in potato anthocyanin synthesis.

Global status and trends in heart failure with preserved ejection fraction over the period 2009-2020: A bibliometric analysis.

BACKGROUND: Heart failure with preserved ejection fraction (HFpEF) comprises about 50% of the cases of heart failure (HF), but so far there is no effective treatment strategy. This study used bibliometric methods to analyze the scientific literature on HFpEF in 2009 to 2020, and evaluate the global scientific output of HFpEF research, in order to explore the research status and trends in this field. METHODS: Documents about the HFpEF research published in 2009 to 2020 were retrieved from Science Citation Index Expanded (SCIE) in Web of Science. This study used bibliometrix R-package, VOSviewer, and CiteSpace to conduct the bibliometric analysis. RESULTS: A total of 1971 documents (1508 articles and 283 reviews) were retrieved to construct the local HFpEF literature collection for analysis. The number of annual documents had increased year by year in general, from 24 to 353. Relevant documents were mainly written in English, and mostly focused on the field of "Cardiovascular System Cardiology." USA ranked first in the relevant countries/regions with most documents, and the leading affiliation was Mayo Clin. Shah SJ was the most productive author, while Borlaug BA ranked highest among the local cited authors and G-index. Circulation was the most local cited source, while Eur J Heart Fail published the most documents and was rated as the top source in terms of G-index. "Paulus WJ, 2013, J Am Coll Cardiol" was the top local cited document within the local HFpEF literature collection, while "Owan TE, 2006, New Engl J Med" outside the local HFpEF literature collection was the most local cited reference. The keywords such as "mortality," "dysfunction," "diagnosis," "outcomes," and "diastolic dysfunction" were most frequent, while "hemodynamics," "comorbidity," "myocardial infarction," "inflammation," and "phenotype" indicated research frontiers or emerging trends. Furthermore, this study also found some deeper bibliometric relationships through bibliographic networks. CONCLUSIONS: Due to the multi-dimensional bibliometric analysis, this study shows a wide view of scientific productivity related to HFpEF, and provides valuable guidance for researchers interested in HFpEF, assisting them in understanding the research status, identifying potential collaborators, discovering research hotspots and frontiers, and conducting more in-depth research.

PCC0208027, a novel tyrosine kinase inhibitor, inhibits tumor growth of NSCLC by targeting EGFR and HER2 aberrations.

PCC-0208027 is a novel tyrosine kinase inhibitor that has a strong inhibitory effect on epidermal growth factor receptor (EGFR)- or HER2-driven cancers. The aim is to assess the anti-tumor activity of PCC0208027 and related mechanisms in non-small cell lung cancer (NSCLC). We examined the activity of PCC0208027 on various mutated EGFRs, HER2, and HER4. MTT assays, flow cytometry, and Western blotting were used to examine the effects of PCC0208027 on NSCLC cells with different genetic characteristics and relevant molecular mechanisms. Nude mouse xenograft models with HCC827, NCI-H1975, and Calu-3 cells were used to evaluate the in vivo anti-tumor activity of PCC0208027. Results showed that PCC0208027 effectively inhibited the enzyme activity of EGFR family members, including drug-sensitive EGFR mutations, acquired drug-resistant EGFR T790M and EGFR C797S mutations, and wild-type (WT) HER2. PCC0208027 blocked EGFR phosphorylation, thereby downregulating downstream PI3K/AKT and MAPK/ERK signaling pathways and inducing G0/G1 arrest in NSCLC cells. PCC0208027 inhibited tumor growth in mouse xenograft models of HCC827, NCI-H1975, and Calu-3 cells. In summary, our findings suggest that PCC0208027 has the potential to become an oral antineoplastic drug for NSCLC treatment and is worthy of further development.

Screening and identification of multiple constituents and their metabolites of Zhi-zi-chi decoction in rat urine and bile by ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry.

Zhi-zi-chi decoction (ZZCD) is a classical formula widely used in Chinese clinical application. In the present study, a novel and efficient strategy has been developed for screening and identification of multiple constituents and their metabolites of ZZCD using ultra-high-performance liquid chromatography combined with triple time-of-flight mass spectrometry. The novel approach of an online data acquisition method dependent on multiple mass defect filter and dynamic background subtraction is combined with multiple data processing techniques. First, a total of 109 potential bioactive compounds were detected in ZZCD. Based on the same instrumental conditions, 100 compounds were found in rat biofluids after oral administration of ZZCD, including 61 original compounds of ZZCD as well as 39 metabolites. Conjugations with sulfate, glucuronate and amino acids were found as the predominant metabolic reaction of ZZCD. As more xenobiotics were detected in urine than those in bile were, it demonstrated that multiple components of ZZCD have undergone comprehensive renal excretion. This study reported the urinary and biliary excretion in rats after oral administration of ZZCD for the first time. The present study expands our knowledge about the constituents and metabolism of ZZCD, which could be very useful for further pharmacological and clinical studies of ZZCD.

(-)-SCR1693 Protects against Memory Impairment and Hippocampal Damage in a Chronic Cerebral Hypoperfusion Rat Model.

Chronic cerebral hypoperfusion (CCH) is one of the most common causes of vascular dementia (VaD) and is recognised as an etiological factor in the development of Alzheimer's disease (AD). CCH can induce severe cognitive deficits, as assessed by the water maze task, along with neuronal loss in the hippocampus. However, there are currently no effective, approved pharmacological treatments available for VaD. In the present study, we created a rat model of CCH using bilateral common carotid artery occlusion and found that (-)-SCR1693, a novel compound, prevented rats from developing memory deficits and neuronal damage in the hippocampus by rectifying cholinergic dysfunction and decreasing the accumulation of the phospho-tau protein. These results strongly suggest that (-)-SCR1693 has therapeutic potential for the treatment of CCH-induced VaD.

Protective effect of carbachol postconditioning on hypoxia/reoxygenation-induced injury in human gastric epithelial cells.

AIMS: We investigated the protective effects of carbachol postconditioning (CAR-P) on acute gastric mucosal injury induced by hypoxia/reoxygenation (H/R) and its possible mechanisms. MAIN METHODS: Cell viability was detected by methyl thiazolyl tetrazolium (MTT). The apoptotic cells were examined by Hoechst 33258 staining. Flow cytometric analysis, lactate dehydrogenate (LDH) release assay, immunocytochemistry, and western blotting were used to investigate the effects of CAR-P on acute gastric mucosal injury induced by H/R. The model of H/R was established by hypoxia induction(94% N2+1% O2+5% CO2 for 2 h) and reoxygenation (normoxic condition for 4 h, 8 h and 16 h). KEY FINDINGS: Our study observed the protective effect of carbachol postconditioning on H/R-induced injury in human gastric epithelial cell lines (hGES-1) cells, which is achieved by direct activation of vanilloid receptor subtype 1 (VR1) and production of calcitonin gene-related peptide (CGRP), and in the inhibition of cell apoptosis. In the study, we demonstrate that CAR-P has protective effects on the H/R-induced injury in hGES-1 cells, and these effects are associated with cholinergic muscarinic receptors (CMR), VR1, and extracellular signal-regulated kinase (ERK) signaling pathway. SIGNIFICANCE: Our findings might provide a new and improved understanding of CAR-P function and an effective treatment strategy for acute gastric mucosal injury induced by H/R.

Protection of pyruvate against glutamate excitotoxicity is mediated by regulating DAPK1 protein complex.

The neuroprotective activity of pyruvate has been confirmed in previous in vivo and in vitro studies. Here, we report a novel mechanism that pyruvate prevents SH-SY5Y cells from glutamate excitotoxicity by regulating death-associated protein kinase 1 (DAPK1) protein complex. Our results showed pyruvate regulated DAPK1 protein complex to protect cells by two ways. First, pyruvate induced the dissociation of DAPK1 with NMDA receptors. The disruption of the DAPK1-NMDA receptors complex resulted in a decrease in NMDA receptors phosphorylation. Then the glutamate-stimulated Ca2+ influx was inhibited and intracellular Ca2+ overload was alleviated, which blocked the release of cytochrome c and cell death. In addition, increased Bcl-xL induced by pyruvate regulated Bax/Bak dependent death by inhibiting the release of cytochrome c from the mitochondrial inter-membrane space into the cytosol. As a result, the cytochrome c-initiated caspase cascade, including caspase-3 and caspase-9, was inhibited. Second, pyruvate promoted the association between DAPK1 and Beclin-1, which resulted in autophagy activation. The autophagy inhibitor 3-methyladenine reversed the protection afforded by pyruvate. Furthermore, the attenuation of mitochondrial damage induced by pyruvate was partly reduced by 3-methyladenine. This suggested autophagy mediated pyruvate protection by preventing mitochondrial damage. Taken together, pyruvate protects cells from glutamate excitotoxicity by regulating DAPK1 complexes, both through dissociation of DAPK1 from NMDA receptors and association of DAPK1 with Beclin-1. They go forward to protect cells by attenuating Ca2+ overload and activating autophagy. Finally, a convergence of the two ways protects mitochondria from glutamate excitotoxicity, which leads to cell survival.

[Systematic review of resuscitation needling technique for wind stroke].

OBJECTIVE: To make a systematic evaluation on therapeutic effect and safety of resuscitation needling technique in treatment of stroke. METHODS: Cochrane systematic assessment was adopted, computerized as well as manual retrieval methods were applied. Meta analyses were conducted by using Review Manager 5.1 software on randomized controlled trial (RCT) and quasi-randomized controlled clinical trials (q-RCT) which complied with the standard. 20 articles and 2809 patients of stroke were included. RESULTS: Therapeutic effect of resuscitation needling technique on enhancing the motor function of human body, improving functional deficiency of nerves and living ability were all better than that of the control group. CONCLUSION: The resuscitation needling technique has good effect on stroke. However, the quality of inclusive literatures is comparatively low. Therefore, more large-sample high-quality RCTs are expected for further studies.

Antitumor effect and toxicity of Lipusu in rat ovarian cancer xenografts.

Paclitaxel has yielded superior therapeutic effects in treating ovarian cancer after intraperitoneal (i.p.) injection. However, the dose-limiting toxicity of Cremophor-based paclitaxel was severe abdominal pain, likely caused by the excipients (Cremophor/ethanol). Lipusu, a paclitaxel liposome, has been widely applied for the treatment of ovarian cancer by intravenous administration in China. In order to find potential benefits of i.p. administration of Lipusu, we suppose that Lipusu could modulate paclitaxel toxicity without affecting antitumor activity compared with Cremophor-based paclitaxel (PTX). Antitumor effects, bone marrow toxicity, cardiotoxicity and biodistributions in NuTu19 ovarian cancer-bearing rats, as well as the abdominalpain in normal mice were evaluated. Lipusu exerted similar antitumor effects similar to PTX, but much lower bone marrow toxicity and cardiotoxicity. Furthermore, Lipusu exhibited similar plasma drug exposure, higher exposure in tumor and pelvic lymph nodes and lower exposure in bone marrow and heart compared with PTX. Additionally, Lipusu induced notably lighter abdominalpain than PTX. These data suggested that Lipusu has similar antitumor effect and superior lymphatic targeting with reduced toxicities compared with PTX via i.p. route, which could be related with altered biodistributions. Therefore, Lipusu could be attractive for further evaluation of treating ovarian cancer by i.p. administration in clinic.

Antidepressant-like effects of the extract from Cimicifuga foetida L.

ETHNOPHARMACOLOGICAL RELEVANCE: Cimicifuga foetida L., a traditional Chinese medicine, has been developed for the treatment of perimenopausal symptoms including depression in China (Brand name: XIMINGTING(®), XMT). The primary active constituents are believed to be the triterpene glycosides. Nevertheless, there are no studies about the antidepressant-like effects of XMT in rodents. AIMS OF THE STUDY: The present study aimed to evaluate antidepressant-like effects of XMT. MATERIALS AND METHODS: Antidepressant-like activity of XMT was studied using forced swimming test (FST) and tail suspension test (TST) in female mice, as well as chronic mild stress (CMS) procedure in female rats. In addition, 5-hydroxytryptophan (5-HTP)-induced head-twitch test and yohimbine toxicity potentiation test in female mice were conducted to propose the possible serotonergic or noradrenergic mechanisms in the antidepressant-like effects of XMT. In mice, XMT was administrated acutely and for 7 consecutive days (20, 40 and 80 mg/kg/day, p.o.); and in rats for 28 consecutive days (10, 20 and 40 mg/kg/day, p.o.). RESULTS: XMT significantly reduced immobility duration in FST and TST without affecting locomotor activity, increased swimming and climbing durations in FST, and enhanced 5-HTP-induced head-twitch response while did not affect yohimbine-induced lethality in female mice. XMT also normalized the inhibition of sucrose intake and decreased the levels of plasma adrenocorticotropic hormone and serum corticosterone and adrenal gland weight in CMS-treated female rats. CONCLUSIONS: These data indicate XMT processes antidepressant-like properties in rodents, which could be related to its serotonergic and noradrenergic activation and normalization of the hypothalamic-pituitary-adrenal axis.

Protection of ghrelin postconditioning on hypoxia/reoxygenation in gastric epithelial cells.

AIM: To investigate the protective effect and mechanisms of ghrelin postconditioning against hypoxia/reoxygenation (H/R)-induced injury in human gastric epithelial cells. METHODS: The model of H/R injury was established in gastric epithelial cell line (GES-1) human gastric epithelial cells. Cells were divided into seven groups: normal control group (N); H/R postconditioning group; DMSO postconditioning group (DM); ghrelin postconditioning group (GH); D-Lys3-GHRP-6 + ghrelin postconditioning group (D + GH); capsazepine + ghrelin postconditioning group (C + GH); and LY294002 + ghrelin postconditioning group (L + GH). 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to detect GES-1 cell viability. Hoechst 33258 fluorochrome staining and flow cytometry were conducted to determine apoptosis of GES-1 cells. Spectrophotometry was performed to determine release of lactate dehydrogenate (LDH). Protein expression of Bcl-2, Bax, Akt, and glycogen synthase kinase (GSK)-3ß was determined by western blotting. Expression of vanilloid receptor subtype 1 (VR1), Akt and GSK-3ß was observed by immunocytochemistry. RESULTS: Compared with the H/R group, cell viability of the GH group was significantly increased in a dose-dependent manner (55.9% ± 10.0% vs 69.6% ± 9.6%, 71.9% ± 17.4%, and 76.3% ± 13.3%). Compared with the H/R group, the percentage of apoptotic cells in the GH group significantly decreased (12.38% ± 1.51% vs 6.88% ± 0.87%). Compared with the GH group, the percentage of apoptotic cells in the D + GH group, C + GH group and L + GH groups significantly increased (11.70% ± 0.88%, 11.93% ± 0.96%, 10.20% ± 1.05% vs 6.88% ± 0.87%). There were no significant differences in the percentage of apoptotic cells between the H/R and DM groups (12.38% ± 1.51% vs13.00% ± 1.13%). There was a significant decrease in LDH release following ghrelin postconditioning compared with the H/R group (561.58 ± 64.01 U/L vs 1062.45 ± 105.29 U/L). There was a significant increase in LDH release in the D + GH, C + GH and L + GH groups compared with the GH group (816.89 ± 94.87 U/L, 870.95 ± 64.06 U/L, 838.62 ± 118.45 U/L vs 561.58 ± 64.01 U/L). There were no significant differences in LDH release between the H/R and DM groups (1062.45 ± 105.29 U/L vs 1017.65 ± 68.90 U/L). Compared with the H/R group, expression of Bcl-2 and Akt increased in the GH group, whereas expression of Bax and GSK-3ß decreased. Compared with the GH group, expression of Bcl-2 decreased and Bax increased in the D + GH, C + GH and L + GH groups, and Akt decreased and GSK-3ß increased in the L + GH group. The H/R group also upregulated expression of VR1 and GSK-3ß and downregulated Akt. The number of VR1-positive and Akt-positive cells in the GH group significantly increased, whereas the number of GSK-3ß-positive cells significantly decreased. These effects of ghrelin were reversed by capsazepine and LY294002. CONCLUSION: Ghrelin postconditioning protected against H/R-induced injury in human gastric epithelial cells, which indicated that this protection might be associated with GHS-R, VR1 and the PI3K/Akt signaling pathway.

Human IL18-IL2 fusion protein as a potential antitumor reagent by enhancing NK cell cytotoxicity and IFN-γ production.

PURPOSE: To obtain a human IL18-IL2 fusion protein by genetic engineering methods and investigate its antitumor activity and mechanism in tumor-bearing mouse models. METHODS: Human IL-18, IL-2 and the fusion gene IL18-IL2 were obtained by PCR, inserted into pBV220 vector and expressed in BL21 (DE3) by 42 °C heat induction. Purified proteins were analyzed by SDS-PAGE and Western blot. The biological activity of IL18-IL2 was first determined by its ability to augment IFN-γ production in PBMCs. Then tumor-bearing mouse models of mouse Lewis lung carcinoma (LLC), human large cell lung carcinoma (NCI-H460) and human colorectal carcinoma (HCT-116) were established to investigate the antitumor activity and mechanism of IL18-IL2. RESULTS: IL18-IL2 was confirmed by Western blot, and its molecular weight was about 34.5 kDa. IL18-IL2 could significantly enhance production of IFN-γ in PBMCs in vitro and induce significant tumor regression in tumor-bearing mouse models of LLC, NCI-H460 and HCT-116 than that of IL-18 and IL-2 separately or combination using. In the mice bearing HCT-116 and LLC, IFN-γ concentrations and natural killer cell cytotoxicity were highly enhanced by IL18-IL2. Anti asialo GM1 could reduce natural killer cell cytotoxicity, production of IFN-γ, and regression of LLC tumor aroused by IL18-IL2. CONCLUSIONS: These results suggested the IL18-IL2 fusion protein showed a synergetic effect on tumor regression, which was related to the great ability of IL18-IL2 in enhancing IFN-γ production and natural killer cell cytotoxicity. The fusion protein was a potential antitumor reagent in cancer immunotherapy.

Escin, a natural mixture of triterpene saponins, exhibits antitumor activity against hepatocellular carcinoma.

Escin, a mixture of triterpene saponins extracted from Aesculus wilsonii Rehd., was used to analyze the antitumor effect in hepatocellular carcinoma in vivo and in vitro. At a dose of 2.8 mg/kg, escin had a rather high inhibition ratio (43.5 %) on mice H22 tumor growth in vivo. The results of the SRB cell viability assay showed that escin could induce significant concentration- and time-dependent inhibition of HepG (2) cell viability. Disruption of the G (1)/S phase of cell cycle progression accompanied by the induction of apoptosis were also observed in HepG (2) cells following escin treatment. The results of pulse-field gel electrophoresis and Western blot analysis show the induction of caspase-independent apoptosis by escin. This study provides evidence that escin induces cell cycle checkpoint arrest and caspase-independent cell death in HepG (2) cells, in support of its efficacious potential as a chemopreventive agent.