Regulation of c-Jun N-Terminal Protein Kinase (JNK) Pathway in Apoptosis of Endothelial Outgrowth Cells Induced by Asymmetric Dimethylarginine.

BACKGROUND Endothelial outgrowth cells (EOCs) are terminal endothelial progenitor cells (EPCs). Asymmetric dimethylarginine (ADMA) has been identified as a novel risk factor for cardiovascular diseases. Our aim in the present study was to investigate the effect of regulation of asymmetric dimethylarginine (ADMA) on EOCs apoptosis and to explore the underlining mechanisms of c-Jun N-terminal protein kinase (JNK) pathway in the process. MATERIAL AND METHODS EOCs were harvested from umbilical cord blood and obtained by using density gradient centrifugation and adhesive culture methods. Endothelial characteristics were identified by immunohistochemistry and fluorescence staining. EOCs were treated with different concentrations of ADMA and detected by flow cytometry. After JNK specific inhibitor (SP600125) was added, EOCs apoptosis protein expressions were measured by Western blot analysis. Proliferation, migration, and vascularization were detected by CCK-8 assay, wound healing assay, and tube-like formation assay, respectively. RESULTS EOCs were successfully extracted from umbilical cord blood and different concentrations of ADMA aggravated EOCs apoptosis. ADMA distinctly activates the phosphorylation activity of JNK. Supplementation of JNK-specific inhibitor (SP600125) decreased expression of Bax and cleaved caspase 3/9, and alleviated ADMA-induced apoptosis. SP600125 also promoted angiogenesis viability. CONCLUSIONS The JNK pathway participates in the apoptosis-promoting process of EOCs, and targeted inhibition of the JNK pathway can alleviate ADMA-induced injury, which I s the potential underlying mechanism of vascular endothelium injury in ischemic stroke.

Immunomodulation of adipose-derived mesenchymal stem cells on peripheral blood mononuclear cells in colorectal cancer patients with COVID-19.

BACKGROUND: Accumulating evidence has shown that adipose tissue-derived mesenchymal stem cells (ADSCs) are an effective therapeutic approach for managing coronavirus disease 2019 (COVID-19); however, further elucidation is required to determine their underlying immunomodulatory effect on the mRNA expression of T helper cell-related transcription factors (TFs) and cytokine release in peripheral blood mononuclear cells (PBMCs). AIM: To investigate the impact of ADSCs on the mRNA expression of TFs and cytokine release in PBMCs from colorectal cancer (CRC) patients with severe COVID-19 (CRC+ patients). METHODS: PBMCs from CRC+ patients (PBMCs-C+) and age-matched CRC patients (PBMCs-C) were stimulated and cultured in the presence/absence of ADSCs. The mRNA levels of T-box TF TBX21 (T-bet), GATA binding protein 3 (GATA-3), RAR-related orphan receptor C (RORC), and forkhead box P3 (FoxP3) in the PBMCs were determined by reverse transcriptase-polymerase chain reaction. Culture supernatants were evaluated for levels of interferon gamma (IFN-γ), interleukin 4 (IL-4), IL-17A, and transforming growth factor beta 1 (TGF-ß1) using an enzyme-linked immunosorbent assay. RESULTS: Compared with PBMCs-C, PBMCs-C+ exhibited higher mRNA levels of T-bet and RORC, and increased levels of IFN-γ and IL-17A. Additionally, a significant decrease in FoxP3 mRNA and TGF-ß1, as well as an increase in T-bet/GATA-3, RORC/FoxP3, IFN-γ/IL-4, and IL-17A/TGF-ß1 ratios were observed in PBMCs-C+. Furthermore, ADSCs significantly induced a functional regulatory T cell (Treg) subset, as evidenced by an increase in FoxP3 mRNA and TGF-ß1 release levels. This was accompanied by a significant decrease in the mRNA levels of T-bet and RORC, release of IFN-γ and IL-17A, and T-bet/GATA-3, RORC/FoxP3, IFN-γ/IL-4, and IL-17A/TGF-ß1 ratios, compared with the PBMCs-C+alone. CONCLUSION: The present in vitro studies showed that ADSCs contributed to the immunosuppressive effects on PBMCs-C+, favoring Treg responses. Thus, ADSC-based cell therapy could be a beneficial approach for patients with severe COVID-19 who fail to respond to conventional therapies.

[SVM-based qualitative analysis of Muscat Hamburg wine produced in Tianjin region].

The purpose was to achieve the identification of Muscat Hamburg wines produced in Tianjin region through scanning and analyzing dry white wine samples of different grape varieties and regions by infrared spectroscopy technology. A support vector machine (SVM) based method was introduced to analyze infrared spectra of dry white wines. The pretreatment processes of the IR spectra were also elaborated, including baseline adjustment, noise Elimination, standard normalization and eliminating the main component of abnormal sample points. The authors selected great quantity of dry white wine samples of different grape regions including 511 Muscat Hamburg wine samples, 438 Italian Riesling wine samples, 307 Chardonnay wine samples, 29 Ugni Blanc wine samples, 44 Rkatsiteli wine samples, 31 longan wine samples and 79 ZeHong wine samples. According to different classification problems, 80% of IR spectra of the wine samples were used to establish discrimination models with SVM-based method, and the remaining 20% of IR spectra were used for the validation of the discrimination models. Experimental results showed that the proposed method is effective, since high classification accuracy, identification rate and rejecting rate were achieved: over 97% for the white wine samples of different grape varieties, meanwhile over 98% for the Muscat Hamburg wine samples produced in different regions. So the method developed in this paper played a good role in the qualitative classification and discrimination of Muscat Hamburg wines produced in Tianjin region. This novel method has a considerable potential and a rosy application future due to the expeditiousness, stability and easy-operation of FTIR method, as well as the veracity and credibility of SVM method.

Reproductive Outcomes of Single Embryo Transfer in Women with Previous Cesarean Section.

The aim of this study is to evaluate the reproductive outcomes of patients with previous cesarean section (CS) undergoing single-embryo transfer (SET). A total of 5479 patients previously underwent CS or vaginal delivery (VD) were included. The patients with previous CS included single/double cleavage (SCT/DCT), single/double blastocyst stage embryo transfer (SBT/DBT). The comparison of reproductive outcomes between CS and VD, SET, and double-embryo transfer of CS was conducted. The main outcome measures included clinical pregnancy rate (CPR), multiple pregnancy rate (MPR), live birth rate (LBR), and cumulative live birth rate (CLBR). There were no differences in CPR and LBR between patients previously underwent CS and VD, and the preterm birth rate of multiple births in patients underwent CS were significantly higher than underwent VD. As for patients underwent CS, the CPR, MPR, and LBR were lower in patients treated with SCT than DCT in IVF/ICSI cycles. The CPR and LBR of patients treated with SCT were not statistically different compared with DCT and SBT when patients' age were younger than 35 years in FET cycles, and the MPR of patients treated with SCT was lower. The CPR and LBR of patients treated with SCT were lower than DCT/SBT when patients' age were 35 years or older in FET cycles. The CPR, LBR, and CLBR of patients treated with SBT were not lower than DCT in IVF/ICSI-ET, FET, and complete cycles, but the MPR was lower. Reducing the number of embryos transferred should be considered seriously for CS. SCT under the 35 years of FET cycles is feasible. SBT is an effective strategy to reduce MPR of IVF/ICSI, FET, and complete cycles regardless of overall or age stratification.

Acupuncture for migraine without aura: a systematic review and meta-analysis.

BACKGROUND: Migraine without aura (MWoA), the most common type of migraine, has great impacts on quality of life for migraineurs. Acupuncture is used in the treatment and prevention of migraine for its analgesic effects. OBJECTIVE: The aim of this systematic review and meta-analysis is to systematically assess the therapeutic and preventive effect of acupuncture treatment and its safety for MWoA. SEARCH STRATEGY: Nine electronic databases (PubMed, MEDLINE, Cochrane Library, Lilacs, Embase, China National Knowledge Infrastructure (CNKI), Chongqing VIP (CQVIP), Wanfang Data and Chinese Clinical Trial Registry (ChiCTR)) were systematically searched from their beginning through June 2017 using MeSH terms such as "acupuncture, acupuncture therapy, electro-acupuncture, ear acupuncture, acupuncture points, acupuncture analgesia," and "migraine disorders, cluster headache." Manual searching included other conference abstracts and reference lists. INCLUSION CRITERIA: Randomized controlled trials (RCTs) with a clinical diagnosis of MWoA, which were treated with acupuncture versus oral medication or sham acupuncture treatment. DATA EXTRACTION AND ANALYSIS: Two evaluators screened and collected literature independently; they extracted information on participants, study design, interventions, follow-up, withdrawal and adverse events and assessed risk of bias and quality of the acupuncture intervention. The primary outcomes were frequency of migraine (FM) and number of migraine days (NM). Secondary outcomes included the visual analogue scale (VAS) score, effective rate (ER) and adverse events. Pooled estimates were calculated as mean difference (MD) with 95% confidence interval (CI) for continuous data and relative risk (RR) with 95% CI for dichotomous data. RESULTS: Overall, 14 RCTs including 1155 participants were identified. The analysis found that acupuncture had a significant advantage over medication in reducing FM (MD = -1.50; 95% CI: -2.32 to -0.68; P < 0.001) and VAS score (MD = 0.97; 95% CI: 0.63-1.31; P < 0.00001) and had a higher ER (RR = 1.30; 95% CI: 1.16-1.45; P < 0.00001). Acupuncture also had a significant advantage over sham acupuncture in the decrease of FM (MD = -1.05; 95% CI: -1.75 to -0.34; P = 0.004) and VAS score (MD = -1.19; 95% CI: -1.75 to -0.63; P < 0.0001). Meanwhile, acupuncture was more tolerated than medication because of less side effect reports (RR = 0.29; 95% CI: 0.17-0.51; P < 0.0001). However, the quality of evidence in the included studies was mainly low (to very low), making confidence in the FM and VAS score results low. CONCLUSION: Our meta-analysis shows that the effectiveness of acupuncture is still uncertain, but it might be relatively safer than medication therapy in the treatment and prophylaxis of MWoA. Further proof is needed.

Effect of Bcl-2 and Bax on survival of side population cells from hepatocellular carcinoma cells.

AIM: To understand the role and significance of side population (SP) cells from hepatocellular carcinoma (HCC) in hepatocarcinogenesis, development, relapse and metastasis, we simulated the denutrition conditions that cancer cells experience in clinical therapy, observed the different anti-apoptosis ability of SP cells and non-SP cells under such conditions, and established the possible effects of P53, Bcl-2 and Bax on survival of SP cells. METHODS: We used flow cytometry to analyze and sort the SP and non-SP cells in established HCC lines MHCC97 and hHCC. We evaluated cell proliferation by methyl thiazolyl tetrazolium (MTT) assay and investigated the expression of p53, bcl-2 and bax genes during denutrition, by RT-PCR and immunofluorescence staining. RESULTS: The percentage of SP cells in the two established HCC lines was 0.25% and 0.5%, respectively. SP cells had greater anti-apoptosis and proliferation ability than non-SP cells. Expression of Bcl-2 and Bax in SP and non-SP cells differed during denutrition. The former was up-regulated in SP cells, and the latter was up-regulated in non-SP cells. CONCLUSION: It may be that different upstream molecules acted and led to different expression levels of Bcl-2 and Bax in these two cell lines. There was a direct relationship between up-regulation of Bcl-2 and down-regulation of Bax and higher anti-apoptosis ability in SP cells. It may be that the existence and activity of SP cells are partly responsible for some of the clinical phenomena which are seen in HCC, such as relapse or metastasis. Further research on SP cells may have potential applications in the field of anticancer therapy.