Lead exposure induces neuronal apoptosis via NFκB p65/RBBP4/Survivin signaling pathway.

Lead (Pb), as a heavy metal that is easily exposed in daily life, can cause damage to various systems of body. Apoptosis is an autonomous cell death process regulated by genes in order to maintain the stability of internal environment, which plays an important role in the development of nervous system. RB binding protein 4 (RBBP4) is one of the core histone binding subunits and is closely related to the apoptosis process of nervous system cells. However, it is not known whether RBBP4 can regulate neuronal apoptosis in lead-exposed environments. We exposed PC12 cells to 0 µM (control group), 1 µM, and 100 µM PbAc for 24 h to obtain cell samples. The female rats ingested drinking water containing 0, 0.5 g/L, and 2.0 g/L PbAc from the first day of pregnancy to three weeks after delivery to obtain hippocampal tissue samples from mammary rats. The results of TUNEL showed that lead exposure promoted the onset of apoptosis in cells and hippocampus. The mRNA and protein levels of the apoptosis-related protein Survivin were significantly reduced in the lead-exposed group compared to the control group. In addition, we found that lead exposure reduces the mRNA and protein levels of RBBP4 in PC12 cells and hippocampus, and increases the mRNA and protein levels of NFκB p65. Moreover, inhibiting NFκB p65 can reverse the decrease in RBBP4 expression in the lead exposure model. Overexpression of RBBP4 increased Survivin expression and reduced apoptosis induced by lead exposure. This suggests that lead exposure induces apoptosis through the NFκB p65/RBBP4/Survivin signaling pathway.

Risk of antiangiogenic adverse events in metastatic colorectal cancer patients receiving aflibercept in combination with chemotherapy: A meta-analysis.

BACKGROUND: Aflibercept has been approved for the treatment of metastatic colorectal cancer for more than a decade, but its antiangiogenesis adverse effect profile during treatment remains unclear. This study is conducted to systematically review the risk of antiangiogenic adverse events in patients with metastatic colorectal cancer receiving aflibercept plus chemotherapy. METHODS: We searched databases, including PubMed, Embase and the Cochrane Library up to September 9, 2021. Relevant randomized controlled trials (RCTs) and single-arm studies were included in the review. Statistical analyses were performed using R to calculate the summary incidence rate of antiangiogenic-related adverse events, odds ratios and 95% CIs. Heterogeneity among the included studies was assessed by subgroup analysis. Publication bias analysis and sensitivity analysis were performed to confirm the reliability of the results. RESULTS: A total of 2889 patients from 10 studies met the inclusion criteria. The quality of the included studies was evaluated as qualified for further quantitative synthesis. In part of single-arm studies, the occurrence rates were 44.2% (95%CI, 39.7-48.7%) for hypertension, 31.3% (95% CI, 19.3-43.3%) for proteinuria, 27.3% (95%CI, 21.2-33.4%) for epistaxis, 22.5% (95%CI, 7.8-37.3%) for hemorrhage events, 8.0% (95%CI, 2.0-14 .0%) for venous thromboembolic event in all grades and 22.6% (95%CI, 19.1-26.2%) for grade III/IV hypertension, 7.4% (95%CI, 6.2-8.5%) for grade III/IV proteinuria. In part of RCT, compared to its counterpart, aflibercept containing arm was associated with the increased incidence rate in hypertension (OR:6.30, 95%CI: 3.49-11.36), proteinuria (OR:4.12, 95%CI: 1.25-13.61), epistaxis (OR:3.71, 95%CI: 2.84-4.85), III/IV hypertension (OR:7.20, 95%CI: 5.23-9.92), III/IV proteinuria (OR:5.13, 95%CI: 3.13-8.41). The funnel plot, Begg test and Egger test were carried out on the primary endpoints, III/IV hypertension rate and III/IV proteinuria rate, the result of which detected no obvious publication bias. No significant difference was observed in subgroup analysis in the primary endpoint between the subgroups stratified by treatment line (firstline or non-firstline), chemotherapy regime (FOLFIRI or others) and study design (RCTs or single-arm trials). CONCLUSION: The available evidence suggests that using aflibercept is associated with an increased risk of antiangiogenic adverse events compared with controls. Further studies are needed to investigate this association. In the appropriate clinical scenario, the use of aflibercept in its approved indications remains justified. However, the results of this study should be interpreted with caution, as some of the evidence comes from single-arm clinical trials.

Cucurbitacin C suppresses the progression of pancreatic ductal adenocarcinoma via inhibition of the cGMP-PKG-VASP axis.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most devastating diseases; it has a considerably poor prognosis and may become the second most lethal malignancy in the next 10 years. Chemotherapeutic resistance is common in PDAC; thus, it is necessary to exploit effective alternative drugs. In recent years, traditional folk medicines and their extracts have shown great potential in cancer treatment. The seed of Lagenaria siceraria (Molina) Standl. is a traditional medicine in Asia. Because of its analgesic effects and ability to reduce swelling, it is often used as an adjuvant treatment for abdominal tumors. Cucurbitacin compounds are extracts abundant in Lagenaria siceraria (Molina) Standl. Here, we found that cucurbitacin C (CuC), a member of the cucurbitacin family, has apparent anti-PDAC therapeutic properties. CuC decreased the viability and suppressed the proliferation of PDAC cells in a time- and dose-dependent manner. Further studies revealed that CuC inhibited cell migration and invasion by inhibiting epithelial-mesenchymal transition (EMT). In addition, G2/M arrest was induced, and the apoptotic pathway was activated. Transcriptomic and bioinformatic analyses showed that CuC inhibited the cGMP-PKG-VASP axis, increasing the content of cGMP to restore tumor characteristics. The antitumor activity of CuC in vivo was verified through animal experiments, and no obvious side effects were observed. Overall, our study indicates a candidate therapeutic compound for PDAC that is worthy of further development.

Efficacy, safety, and cost-effectiveness analysis of aflibercept in metastatic colorectal cancer: A rapid health technology assessment.

Background: Metastatic colorectal cancer (mCRC) imposes a heavy tumor burden worldwide due to limited availability of therapeutic drugs. Aflibercept, a kind of recombinant protein of the anti-vascular endothelial growth factor (VEGF) family, has been approved in clinical application among mCRC patients since 2012. A comprehensive analysis of the efficacy, safety, and cost-effectiveness of aflibercept in mCRC treatment is necessary. Objective: To evaluate the efficacy, safety, and cost-effectiveness of aflibercept for the treatment of mCRC in order to provide a decision-making reference for the selection of targeted drugs for second-line treatment of mCRC in Hong Kong, Macao, and Taiwan regions of China and the selection of new drugs for medical institutions in these regions. Methods: A systematic retrieve on databases including PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang, and Weipu, as well as relevant websites and databases of health technology assessment including the National Institute of Health and Clinical Optimization, Centre for Evaluation and Communication at the University of York, and the Canadian Agency for Medicines and Health Technology, was conducted. The literature was screened according to the inclusion and exclusion criteria, and data were extracted and analyzed by two authors, while the quality of the literature was assessed. Results: Finally, we included two HTA reports, 11 systematic reviews/meta-analyses, and two cost-effectiveness studies in the rapid health technology assessment. For mCRC patients receiving second-line treatment, aflibercept combined with FOLFIRI significantly increased progression-free survival (PFS) and overall survival (OS) and the objective response rate (ORR) also improved, compared with folinic acid + fluorouracil + irinotecan (FOLFIRI). In terms of safety, mCRC patients who received aflibercept combined with FOLFIRI therapy had a higher incidence of grade 3-4 adverse events than those who received FOLFIRI alone, including anti-VEGF-related adverse events (hypertension, hemorrhagic events, and proteinuria) and chemotherapy-related adverse events (diarrhea, weakness, stomatitis, hand-foot syndrome, neutropenia, and thrombocytopenia). In terms of cost-effectiveness, two economic studies conducted in the United Kingdom and Japan, respectively, found that compared with FOLFIRI, aflibercept combined with FOLFIRI had no cost-effectiveness advantage in mCRC patients receiving second-line treatment. Conclusion: Compared with FOLFIRI treatment, aflibercept combined with FOLFIRI for the second-line treatment of mCRC patients has better efficacy, worse safety, and is not cost-effective. More high-quality clinical studies are required for further exploration of aflibercept's clinical value. Medical institutions in Hong Kong, Macao, and Taiwan regions of China should be cautious when using or introducing aflibercept plus FOLFIRI as a mCRC treatment.

Linderalactone Suppresses Pancreatic Cancer Development In Vitro and In Vivo via Negatively Regulating PI3K/AKT Signaling Pathway.

Linderalactone is one of the main extracts of Linderae Radix, which is widely used in traditional Chinese medicine. There have been few studies on the antitumor effect of linderalactone in the past. In this study, we explored the anti-pancreatic cancer activity of linderalactone in vitro and in vivo. The results showed that linderalactone inhibited the proliferation of pancreatic cancer cells in a time- and dose-dependent manner. Cell migration and invasion were significantly inhibited by linderalactone. The cell cycle was arrested in the G2/M phase, and the expression levels of cell cycle-associated proteins changed significantly with linderalactone treatment. In addition, linderalactone induced cell apoptosis and altered the expression of apoptotic markers, such as caspase 3 and PARP1. Mechanistically, linderalactone suppressed the PI3K/AKT signaling pathway by downregulating the phosphorylation of PI3K and AKT. The xenograft study results were consistent with the in vitro results, and there was no obvious chemical toxicity. Thus, our research demonstrated that linderalactone exhibits antitumor activity against pancreatic cancer and may be developed as a potential anti-pancreatic cancer agent in the future.

Psychometric Properties of the Fertility Intention Scale among Patients with Breast Cancer of Childbearing Age in Mainland China.

Objective: There are no valid assessment instruments assessing fertility intention among breast cancer survivors in mainland China. Therefore, this study aims to examine the psychometric properties of the Taiwanese version of the Fertility Intention Scale (FIS) among female patients with breast cancer of childbearing age in mainland China. Methods: Two hundred and sixty-four female patients with breast cancer of childbearing age were recruited from two tertiary hospitals in Tianjin and Baoding, of which 32 patients completed the survey twice. Confirmatory factor analysis was adopted to assess construct validity. Correlations between the Reproductive Concerns After Cancer scale and FIS scores were calculated using Spearman correlation for convergent validity. The known-group validity of the FIS was verified using Mann-Whitney U test to compare the FIS scores between patients with or without the intention to conceive. Moreover, reliabilities were examined using Cronbach's alpha and intra-class correlation coefficient. Results: Confirmatory factor analysis showed a good model fit to previous factor structures (χ2/df = 3.19, root mean square error of approximation = 0.091, comparative fitting index = 0.980, Tucker-Lewis index = 0.975), and no FIS item was dropped. The FIS scores were weak negatively correlated with the Reproductive Concerns After Cancer scale scores (r = -0.172, P < 0.01). The convergent validity of FIS was not satisfactory. Differences were noted between patients with or without the intention to conceive (50.62 ± 6.35 vs. 45.98 ± 7.19, P < 0.01). The FIS showed acceptable known-group validity. The internal consistency (Cronbach's α = 0.824) and the test-retest reliability (r = 0.863, P < 0.01) of the FIS were also acceptable. Conclusions: Overall, the FIS provides a comprehensive evaluation of the fertility intention among patients with breast cancer of childbearing age in mainland China. However, the convergent validity was not satisfactory; thus, further revision and validation may be required in the future.

Essential Oils of Gardenia jasminoides J. Ellis and Gardenia jasminoides f. longicarpa Z.W. Xie & M. Okada Flowers: Chemical Characterization and Assessment of Anti-Inflammatory Effects in Alveolar Macrophage.

Alveolar macrophage is the predominant cell type in the lung and is thought to be the major target for anti-inflammatory therapy in chronic obstructive pulmonary disease (COPD). Aromatherapy using natural essential oils with anti-inflammatory effects for inhalable administration is a potential complementary and alternative therapy for COPD treatment. The Gardenia jasminoides flower is famous for its fragrance in East Asia and is used for treating colds and lung problems in folk medicine. Therefore, in the present study, flower essential oils from two main medicinal gardenia varieties (G. jasminoides J. Ellis and G. jasminoides f. longicarpa Z.W. Xie & M. Okada) were extracted by hydro-distillation, and their chemical components were analyzed by GC-MS. The anti-inflammatory effects of the two essential oils and their main ingredients were further studied on lipopolysaccharide (LPS)-induced models in murine alveolar macrophages (MH-S). The results indicated that the chemical constituents of the two gardenia varieties were quite different. Alcohol accounted for 53.8% of the G. jasminoides essential oil, followed by terpenes (16.01%). Terpenes accounted for 34.32% of the G. jasminoides f. longicarpa essential oil, followed by alcohols (19.6%) and esters (13.85%). Both the two gardenia essential oils inhibited the LPS-induced nitric oxide (NO) release and reduced the production of tumor necrosis factor-α (TNF-α) and prostaglandin E2 (PGE2) in the MH-S cells. Linalool and α-farnesene dose-dependently reduced the NO release in the MH-S cells. Linalool and α-farnesene did not affect the PGE2 production but regulated the expression of TNF- α. In addition to linalool and α-farnesene, other components in the gardenia flower essential oils appeared to be able to act as anti-inflammatory agents and influence the PGE2 pathway.

Efficacy and safety of aflibercept plus chemotherapy in metastatic colorectal cancer: A systematic review and PRISMA-Compliant single-arm Meta-Analysis of noncomparative clinical studies and randomized controlled trials.

WHAT IS KNOWN AND OBJECTIVE: Aflibercept, a recombinant protein designed to suppress the vascular endothelial growth factor (VEGF) signalling pathway, has been used in patients with metastatic colorectal cancer (mCRC). We conducted the first meta-analysis to systematically review the efficacy and safety of aflibercept in mCRC. METHODS: PubMed Central/Medline, Embase and cochrane library were systematically searched for randomized controlled trials and single-arm clinical trials on aflibercept plus chemotherapy for the treatment of mCRC through 9 September 2021. RESULTS: Ten studies comprising 2049 patients met the inclusion criteria. The pooled estimate rates were 16.0% for 12mPFS, 64.4% for 12mOS, 32.5% for ORR, 83.5% for DCR, while the rates of III/IV AEs rate were 80.2% respectively. The pooled estimate rates were 16.8% for III/IV diarrhoea, 22.3% for III/IV hypertension, 29.5% for III/IV neutropenia, 7.3% for III/IV proteinuria and 8.6% for III/IV oral mucositis. CONCLUSIONS: Analysis of data from randomized controlled trials(RCT) and single-arm clinical trials confirmed the good efficacy of aflibercept plus chemotherapy in mCRC, while the safety of the treatment is concerning.

Abnormal expression of long non-coding RNA rhabdomyosarcoma 2-associated transcript (RMST) participates in the pathological mechanism of atherosclerosis by regulating miR-224-3p.

Study shows that long non-coding RNA (lncRNA) plays a regulatory role in cardiovascular diseases, and the mechanism of rhabdomyosarcoma 2-associated transcript (RMST) in atherosclerosis (AS) is still unclear. This study aimed to evaluate the expression of RMST and its possible role in the occurrence of AS. RMST and miR-224-3p level in serum and human umbilical vein endothelial cells (HUVECs) were determined by real-time quantitative PCR (RT-qPCR). In vitro atherosclerotic cell model was achieved by treating HUVECs with ox-LDL. Receiver operating characteristic (ROC) curve assessed the diagnostic value of RMST in AS, and Pearson correlation coefficient estimated the correlation of RMST with carotid intima-media thickness (CIMT) and carotid-femoral pulse wave velocity (cfPWV). Cell counting kit-8 (CCK-8) assay and Enzyme-linked immunosorbent assay (ELISA) were performed to evaluate the effect of RMST on cell viability and inflammatory response. The luciferase analysis was used to validate the relationship between RMST and miR-224-3p. The results showed that in serum and HUVECs, RMST levels were increased, while miR-224-3p level was decreased. ROC curve suggested that RMST had clinical diagnostic value for AS. Besides, CIMT and cfPWV were positively correlated with RMST levels, respectively. In HUVECs, RMST-knockdown notably improved the cell viability and inhibited the production of inflammatory factors. Moreover, miR-224-3p was the target of RMST. In conclusion, RMST has the potential to be a diagnostic marker for AS. RMST-knockdown contributes to the enhancement of cell viability and the inhibition of inflammatory response, which may provide new insights into the conquest of AS.

Advancements in lead therapeutic phytochemicals polycystic ovary syndrome: A review.

Polycystic ovary syndrome (PCOS) is one of the most common endocrine diseases in women of reproductive age and features complex pathological symptoms and mechanisms. Existing medical treatments have, to some extent, alleviated the deterioration of PCOS. However, these strategies only temporarily control symptoms, with a few side effects and no preventive effect. Phytochemicals extracted from medicinal herbs and plants are vital for discovering novel drugs. In recent years, many kinds of research have proven that phytochemicals isolated from traditional Chinese medicine (TCM) and medicinal plants show significant potential in preventing, alleviating, and treating PCOS. Nevertheless, compared to the abundance of experimental literature and minimal specific-topic reviews related to PCOS, there is a lack of systematic reviews to summarize these advancements in this promising field. Under this background, we systematically document the progress of bioactive phytochemicals from TCM and medicinal plants in treating PCOS, including flavonoids, polyphenols, and alkaloids. According to the literature, these valuable phytochemicals demonstrated therapeutic effects on PCOS supported by in vivo and in vitro experiments, mainly depending on anti-inflammatory, antioxidation, improvement of hormone disorder and insulin resistance (IR), and alleviation of hyperinsulinemia. Based on the current progress, future research directions should emphasize 1) exploring bioactive phytochemicals that potentially mediate bone metabolism for the treatment of PCOS; 2) improving unsatisfactory bioavailability by using advanced drug delivery systems such as nanoparticles and antibody-conjugated drugs, as well as a chemical modification; 3) conducting in-depth research on the pathogenesis of PCOS to potentially impact the gut microbiota and its metabolites in the evolution of PCOS; 4) revealing the pharmacological effects of these bioactive phytochemicals on PCOS at the genetic level; and 5) exploring the hypothetical and unprecedented functions in regulating PCOS by serving as proteolysis-targeting chimeras and molecular glues compared with traditional small molecule drugs. In brief, this review aims to provide detailed mechanisms of these bioactive phytochemicals and hopefully practical and reliable insight into clinical applications concerning PCOS.

Quantitative Assessment of Abiotic Stress on the Main Functional Phytochemicals and Antioxidant Capacity of Wheatgrass at Different Seedling Age.

The wheat seedlings of 6 days old were daily subjected to ultraviolet irradiation (irradiating for 5, 10, 20, 40, and 60 min/day, respectively), Polyethylene glycol 6000 (5, 10, 15, 20, 25% in 1/2 Hoagland solution, respectively), and salinity solution (10, 25, 50, 100, 200 mM in 1/2 Hoagland solution, respectively), while the control group (CK) was supplied only with the Hoagland solution. The wheatgrass was harvested regularly seven times and the total soluble polysaccharides, ascorbic acid, chlorophyll, total polyphenol, total triterpene, total flavonoid, and proanthocyanins content were tested. The antioxidant capacity was evaluated through 2,2'-azino-bis (3-ethylbenzthia-zoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging ability, and ferric ion reducing power. Technique for order preference by similarity to ideal solution (TOPSIS) mathematical model was adopted to comprehensively assess the functional phytochemicals of the different treatments. The results showed that the accumulation patterns of phytochemicals under abiotic stress were complex and not always upregulated or downregulated. The antioxidant activity and functional phytochemicals content of wheatgrass were significantly affected by both the stress treatments and seedling age, while the latter affected the chemicals more efficiently. The top five highest functional phytochemicals were observed in the 200 mM NaCl treated group on the 21st and 27th day, 25% PEG treated group on the 24th day, 200 mM NaCl treated group on the 24th day, and the group of 40 min/day ultraviolet exposure on 27th day.

The Correlation between Demographical and Lifestyle Factors and Traditional Chinese Medicine Constitution among Macau Elderly Individuals.

OBJECTIVES: To measure the distributed characteristics of Traditional Chinese Medicine (TCM) constitutions, as well as related factors with biased TCM constitutions among the elderly individuals in Macau. METHODS: The elderly individuals from elderly healthcare centers located in Macao Peninsula, Taipa, and Coloane were selected as research samples. The basic information questionnaire and the Constitution in Chinese Medicine Questionnaire (CCMQ) for elderly were employed. Descriptive analysis was applied to illustrate demographical characteristics and TCM constitution distribution. Exploratory factor analysis (EFA) was conducted to explore potential factors influencing biased constitutions, and weight of each variable for constitution was further calculated. RESULTS: A total of 313 participants were recruited. Eighty-six (27.48%) elderly were identified as balanced constitution; others were biased constitutions accounting for 72.52%. Distribution differences related to gender and age were identified among participants with unbalanced constitutions. Four biased constitutions were further analyzed with EFA. For qi-stagnation and yang-deficiency constitutions, three similar factors were determined in the domains of education, sleeping habits, and lifestyle behaviors, successively. Although four factors were identified in phlegm-dampness constitution, the latter two belonged to lifestyle behaviors and the former two were education and sleeping habits. For yin-deficiency constitution, education, tobacco-alcohol consumption, sleeping habits, and exercise were four dimensions of potential influential factors. Tobacco consumption, sleep, and exercise duration weighted the most for qi-stagnation constitution; sleep duration, education level, and sugar-containing beverage intake for phlegm-dampness; length of education, alcohol consumption, and education level for yang-deficiency constitution; and weekly exercise hours, sleep duration, and education level for yin-deficiency constitution. CONCLUSION: The prevalence rate of biased constitutions was relatively high among elderly residents in Macau. Relations between demographical and lifestyle behavioral factors and biased constitutions were identified in this study. Controlling these influential factors might be beneficial for health management of Macau elderly individuals.

Effects of Smoking, and Drinking on Serum Gamma-Glutamyl Transferase Levels Using Physical Examination Data: A Cross-Sectional Study in Northwest China.

PURPOSE: Although drinking and smoking have been associated with gamma-glutamyl transferase (GGT) levels, studies on the effects of smoking and drinking on GGT levels are scarce. The objective of this study was to assess the individual and combined effects of smoking, drinking on GGT levels in the Shaanxi province of Northwest China. PARTICIPANTS AND METHODS: A questionnaire survey was conducted in a population that underwent health examination at the First Affiliated Hospital of Xi'an Jiaotong University and included employees of enterprises or public institutions. The survey was used to collect the baseline characteristics, smoking status, and drinking status of the participants. This information was collected from January 2019 to December 2019. Data related to the physical examinations were exported using the hospital information system (HIS). A linear regression model was employed to explore the effects of smoking and drinking on GGT levels. The restricted cubic spline model was applied to assess the dose-response relationship between amount of smoking, alcohol consumption and GGT levels. RESULTS: A total of 10,177 participants were included in the study. Linear regression indicated that smoking (ß=3.37, 95% CI: 2.57-4.17) and drinking (ß=5.55 L, 95% CI: 4.40-6.71) individually, and collectively (ß=9.30, 95% CI: 7.83-10.76) had a positive effect on GGT levels. The restricted cubic spline presented a linear dose-response relationship between the amount of daily smoking and GGT levels (P for non-linearity=0.148, P for overall association <0.001, OR=2.49, 95% CI: 1.27-4.90), and that between weekly alcohol consumption and GGT levels (P for non-linearity=0.231, P for overall association <0.001, OR=4.79, 95% CI: 1.72-13.32). In case of females, stratified analysis showed that in comparison to the reference group, only current drinkers had a significant effect on GGT levels (OR=3.37, 95% CI: 0.19-6.55). CONCLUSION: Smoking and drinking have a dose-dependent and a synergistic effect on GGT levels. They should be controlled concurrently, especially among males.

PWAR6 interacts with miR­106a­5p to regulate the osteogenic differentiation of human periodontal ligament stem cells.

Human periodontal ligament stem cells (hPDLSCs) associated with bone regeneration serve an important role in the treatment of periodontal disease. Long non­coding RNAs are involved in the osteogenesis of multiple stem cells and can act as a sponge of microRNAs (miRs). The present study aimed to investigate the interaction between Prader Willi/Angelman region RNA 6 (PWAR6) and miR­106a­5p, as well as their influences on the osteogenic differentiation of hPDLSCs. hPDLSCs were isolated and cultured in osteogenic medium (OM) or growth medium (GM) for 7 days prior to transfection with PWAR6 overexpression vector, short hairpin RNA PWAR6 or miR­106a­5p mimic. The expression levels of runt­related transcription factor 2, osteocalcin and bone morphogenetic protein 2 (BMP2) were detected by western blotting and reverse transcription­quantitative PCR (RT­qPCR), and the expression levels of PWAR6, miR­106a­5p and alkaline phosphatase (ALP) were determined by RT­qPCR. ALP activity assays and Alizarin red staining were performed to detect osteogenesis and mineralization, respectively. Luciferase activities of wild­type and mutant PWAR6 and BMP2 were assessed by conducting a dual­luciferase reporter assay. The results indicated that PWAR6 expression was upregulated in OM­incubated hPDLSCs compared with GM­incubated hPDLSCs, and PWAR6 overexpression increased the osteogenic differentiation and mineralization of hPDLSCs compared with the corresponding control group. By contrast, miR­106a­5p expression was downregulated in OM­incubated hPDLSCs compared with GM­incubated hPDLSCs. PWAR6 acted as a sponge of miR­106a­5p and PWAR6 overexpression promoted the osteogenesis of miR­106a­5p mimic­transfected hPDLSCs. BMP2 was predicted as a target gene of miR­106a­5p. Collectively, the results indicated that PWAR6 displayed a positive influence on the osteogenic differentiation of hPDLSCs. The results of the present study demonstrated that the PWAR6/miR­106a­5p interaction network may serve as a potential regulatory mechanism underlying hPDLSCs osteogenesis.

Comprehensive Evaluation of the Effect of Ultraviolet Stress on Functional Phytochemicals of Hulless Barley (Qingke) Grass in Different Growth Times at Vegetative Stage.

The present study was executed to analyze the functional phytochemicals of hulless barley grass grown under different intensities of ultraviolet stress. The wheat seedlings were imposed to 0.5, 1.0, 1.5, 2.0, and 2.5 h ultraviolet radiation and harvested in different times at vegetative stage. Specifically, the contents of total polyphenols, total flavonoids, total triterpenes, total polysaccharides, proanthocyanidins, and chlorophyll were determined and antioxidants capacity was evaluated by OH• and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging ability. A mathematical model (Technique for Order Preference by Similarity to Ideal Solution, TOPSIS) was also employed for the comprehensive evaluation of functional components of hulless barley grass at different growth stages. The results showed that the UV stress could efficiently improve/preserve the contents of total polyphenols, total flavonoids, total triterpenes, total polysaccharides, proanthocyanidins, chlorophyll a, chlorophyll b, and total chlorophyll, as well as the OH• and ABTS scavenging capacity. TOPSIS evaluation revealed that the highest phytochemical contents were yield on the 15th day under 1.0 h ultraviolet treatment.

SP1/TGF­ß1/SMAD2 pathway is involved in angiogenesis during osteogenesis.

The relationship between osteoblasts and angiogenesis is vital for bone regeneration, especially mandibular and maxillary bones. Transforming growth factor ß1 (TGF­ß1) and vascular endothelial growth factor (VEGF) are closely related to angiogenesis; however, the regulatory mechanism between them remains unknown. The present study aimed to reveal this mechanism to provide novel insight for development of potential therapeutic opportunities. Western blotting and reverse transcription­quantitative PCR was used to assess the protein and mRNA expression levels in MC3T3­E1 preosteoblast cells and HUVECs, ELISAs were used to detect the expression levels of secreted VEGF, MTT assays were used to assess the viability of the cells, migratory ability was assessed using Transwell assays, angiogenesis assays were used to analyze the formation of blood vessels, and TGF­ß1 regulation was confirmed using a dual­luciferase reporter assay. The overexpression of specificity protein 1 (SP1) or TGF­ß1 increased VEGF expression levels and secretion, and promoted angiogenesis of co­cultured HUVECs. SP1 also promoted SMAD2 phosphorylation. These effects of SP1 were all reversed by the TGF­ß1 inhibitor. The VEGF inhibitor bevacizumab also reduced the SP1/TGF­ß1/SMAD2 pathway­induced angiogenesis of preosteoblasts. In conclusion, it was demonstrated that SP1 promoted TGF­ß1 expression, activated the SMAD2 pathway and induced VEGF secretion, which may enhance angiogenic processes in preosteoblasts.

Quantitation of estradiol by competitive light-initiated chemiluminescent assay using estriol as competitive antigen.

BACKGROUND: Light-initiated chemiluminescent assays (LICA) are homogeneous assays that are sensitive, specific, and free of separation and washing steps and have high throughput and high precision. METHODS: In this research, we developed a competitive method by LICA to achieve accurate quantification of estradiol (E2) in human serum. E2 competed with estriol (E3) for binding to anti-human E2 antibodies. E3 was linked to biotin via bovine serum albumin as a linker. As this assay used competition between the labeled tracer and the analyte, an increase in E2 concentration will cause a signal decrease. RESULTS: The expected detection range of E2 was 20-5000 pg/mL. The analytical and functional sensitivities were 7.16 and 13.7 pg/mL, respectively. The intra- and inter-assay coefficients of variation were both below 15%, and the recovery rate ranged from 97.5% to 106.8%. The interference rates ranged from -3.6% to 5.4% and met detection requirements for E2 in hyperbilirubinemia, hemolysis, and lipemia in clinical samples. In addition, the cross-reactivity rates between E2 and structural analogs and some reproductive hormones varied from 1.9% to 10.6% which showed that LICA is highly specific for E2. Moreover, our results showed high accordance with the IMMULITE 2000 (y = 0.6695x + 47.92, r2  = .843) and VIDAS systems (y = 1.099x - 821.5, r2  = .9392). CONCLUSION: Our data show that the LICA, which is easy to automate, is a promising technique for quantification of E2 in human serum and could be used for clinical detection.

[Anti-platelet aggregation and anti-thrombotic mechanism of Trichosanthis Fructus combined with aspirin based on network pharmacology].

To explore the anti-platelet aggregation and anti-thrombotic mechanisms of Trichosanthis Fructus combined with aspirin based on network pharmacology and the validation of arteriovenous by pass model in rats. The databases of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Drug Repositioning and Adverse Drug Reaction Chemical-Protein Interactome(DRAR-CPI),Universal Protein Resource(Uniprot) and the Database for Annotation,Visualization,and Integrated Discovery(DAVID) were used to predict protein targets and analyze biological pathway and signal pathway in the combination of Trichosanthis Fructus with aspirin. The effects of pretreatment with Trichosanthis Fructus pellets,aspirin pellets and their combination on thromboxane B2(TXB2),6-keto prostaglandin F1α(6-keto-PGF1α) and cyclic adenosine monophosphate(c AMP) in rat thrombotic model were studied. Through the study of network pharmacology,12 components of aspirin and Trichosanthis Fructus,including hydroxygenkwanin,quercetin and adenosine,were found to show the anti-platelet aggregation and anti-thrombosis mechanisms through9 common protein targets,such as SRC,RAC1,MAPK14,MAPK1,AKT1,and 14 common signaling pathways,such as VEGF signaling pathway. After the intervention with Trichosanthis Fructus pellets combined with aspirin pellets,the vascular endothslia growth factor(VEGF) signaling pathway can be activated to inhibit platelet aggregation and improve vascular endothelial function,and show the anti-platelet aggregation and anti-thrombosis mechanisms,which verify the results of the network pharmacology,and explain the anti-platelet aggregation and anti-thrombotic mechanisms of the combination of Trichosanthis Fructus pellets with aspirin pellets.

Upregulation of lncRNA PlncRNA-1 indicates the poor prognosis and promotes glioma progression by activation of Notch signal pathway.

Prostate cancer-up-regulated long noncoding RNA 1(PlncRNA-1) has been demonstrated to be increased in several cancers, which plays an oncogenic role in the development of cancer. However, the exact functions and molecular mechanism of PlncRNA-1 in the tumorigenesis of glioma has not been studied. In present work, we firstly identified that PlncRNA-1 expression levels were prominently augmented in glioma patient tissues and glioma cell lines compared with adjacent noncancerous tissue and normal cells, respectively. Moreover, Kaplan-Meier survival analysis indicated that glioma patients with high PlncRNA-1 expression had shorter overall survival (OS) and progression-free survival (PFS) than those with low PlncRNA-1 expression. Furthermore, loss-of-function assay showed that PlncRNA-1 knockdown dramatically reduced cell proliferation, colony formation, and promoted apoptosis of glioma cell lines. In addition, overexpression of PlncRNA-1 promoted cell proliferation, stimulated cell colony formation, and inhibited cell apoptosis in NHA cells. Mechanically, our results showed that PlncRNA-1 significantly promoted activation of the Notch signal pathway through regulation of Notch-1, Jag-1, and Hes-1 expression. Collectively, our results implied that lncRNA PlncRNA-1 may exert tumor-promoting role in the development and progression of glioma through modulation of Notch signal pathway, providing a candidate therapeutic target for patients with glioma.

Long noncoding RNA HNF1A-AS1 indicates a poor prognosis of colorectal cancer and promotes carcinogenesis via activation of the Wnt/ß-catenin signaling pathway.

Long non-coding RNAs (lncRNAs) have been identified to play critical roles in tumorigenesis. LncRNA HNF1A-AS1 has been suggested to act as an oncogene and serves as a novel prognostic biomarker for various cancer. However, the biological role and clinical significance of lncRNA HNF1A-AS1 in colorectal cancer (CRC) have yet to be fully elusive. Therefore, the present study was designed to determine the expression of lncRNA HNF1A-AS1 in patients with CRC, the role of lncRNA HNF1A-AS1 in CRC cells, as well as the underlying regulatory mechanisms. Our results indicated that the expression of lncRNA HNF1A-AS1 was significantly upregulated in both CRC tumor tissues and CRC cell lines in comparison with adjacent non-tumor tissues and the human normal colonic epithelial cell line (HcoEpiC). The Kaplan-Meier survival analysis further suggested that high expression of lncRNA HNF1A-AS1 might be an independent prognostic factor for disease-free survival (DFS) and overall survival (OS) in patients with CRC. Moreover, the area under the receiver operating characteristic (ROC) curve for HNF1A-AS1 was up to 0.8714, implying that HNF1A-AS1 had diagnostic significance as it could discriminate tumor tissues from nontumorous tissues. In addition, silencing of lncRNA HNF1A-AS1 abrogated the proliferation of CRC cells by MTS assay and clonogenic assay, arrested cell cycle at G0/G1 stage and reduced the migration and invasion in CRC cells. Finally, we found that decreased expression of lncRNA HNF1A-AS1 suppressed the Wnt/ß-catenin signaling pathway activity by downregulating the expression of ß-catenin,cyclinD1, and c-myc. In conclusion, these findings provide evidence that lncRNA HNF1A-AS1 may be considered as a new prognostic biomarker and therapeutic target in patients with CRC.