[Expression of SQSTM1 in papillary thyroid carcinoma and its influence on invasion, migration and proliferation of papillary thyroid cancer cells].

Objective: To investigate the expression of SQSTM1 in thyroid papillary carcinoma and its influence on the invasion and migration of thyroid papillary carcinoma cells TPC-1. Methods: From April to June 2019, cancer tissues and adjacent tissues of 21 cases with thyroid papillary carcinoma in the First Affiliated Hospital of Zhengzhou University were collected, and the expression of SQSTM1 was detected by RT-qPCR. SQSTM1 knockdown cell line SQSTM1-KD-TPC-1 was constructed in TPC-1 cells by lentivirus transfection. RT-qPCR was used to detect SQSTM1 expression in TPC-1 cells and SQSTM1-KD-TPC-1 cells. The changes of invasion and migration before and after SQSTM1 knockdown in TPC-1 cells were detected by transwell test. The proliferation of TPC-1 and SQSTM1-KD-TPC-1 cells were detected by MTT and clone formation test. RT-qPCR was used to detect the gene expression of proliferation related proteins. Results: The expression of SQSTM1 in papillary thyroid carcinoma tissues was significantly higher than that in normal adjacent tissues, and 76.2%(16/21) of the petients showed high mRNA expression. Knock down SQSTM1 significantly inhibited the ability of tumor proliferation, invasion and migration, and the expression of proliferation-related proteins were significantly decreased (P<0.01), indicating that SQSTM1 was involved in the regulation of proliferation related pathway mechanism. Conclusion: SQSTM1 significantly promotes invasion, migration and proliferation in thyroid papillary cancer cells TPC-1 and may be a potential gene therapy target.

[Study on Cep63 expression and apoptosis of thyroid papillary carcinoma cell lines TPC-1].

Objective: To investigate the effect of centrosomal protein Cep63 on the apoptosis of papillary thyroid carcinoma (PTC) cell lines TPC-1 and underlying mechanism. Methods: With collected PTC tissues and adjacent tissues, Cep63 expression was detected by RT-qPCR and its relationship with clinicopathological factors was analyzed. The experiment included negative control group (NC), low expression group (Cep63(-)) and overexpression group (Cep63(+)), and wild-type TPC-1 cells were transfected with Cep63 lentivirus. The efficiency of Cep63 was detected by western blot (WB) and qRT-PCR. Cell proliferation ability was detected by plate cloning experiment and MTT assay. Cell apoptotic rate was detected by flow cytometry, and expression levels of apoptosis-related proteins were detected by immunohistochemistry and WB. The t-test was used to compare the differences in the means between the two groups, the one-way analysis of variance was used to compare multiple groups, and the chi-square test was used to analyze the association between gene expression levels and pathological factors. Results: Compared with NC group, cell proliferation ability was significantly decreased in Cep63(-) group (3.18±0.07 vs. 2.14±0.09, t=8.54, P<0.01) and significantly increased in Cep63(+) group (3.18±0.07 vs. 3.58±0.10, t=3.21, P<0.05). Apoptotic rates in NC, Cep63 (-) and Cep63 (+) groups were respectively 3.03%±0.24%, 8.66%±0.44% and 1.17%±0.44%, and the flow cytometry showed that the low expression of Cep63 significantly increased the apoptosis TPC-1 cells (F=157.7, P<0.001). Bcl-2 protein expression levels of NC, Cep63 (-) and Cep63 (+) groups were respectively 1.07±0.03, 0.49±0.01 and 1.99±0.09, and BAX protein expression levels of three groups were respectively 0.64±0.02, 1.06±0.01 and 0.21±0.03. WB showed that the expression level of Bcl-2 decreased (F=183.2, P<0.001), while the expression level of BAX was significantly up-regulated (F=283.7, P<0.001). Conclusion: Cep63 may regulate the apoptotic process of TPC-1 cells through Bcl-2/BAX pathway and Cep63 may be a potential oncogene of PTC.

Clinical significance of FSTL 1, Bax, Bcl-2 in acute cerebral infarction and its relationship with hemorrhagic transformation.

OBJECTIVE: Acute cerebral infarction (ACI) is the most common type of acute cerebrovascular disease so far, and its incidence rate has been increasing in recent years. At present, the methods of diagnosing ACI in clinic are extremely complicated, and an effective index that can effectively diagnose ACI is urgently needed in clinic. This study is designed to investigate the clinical significance of Follistatin-like protein 1 (FSTL1), Bax and Bcl-2 in ACI. PATIENTS AND METHODS: A total of 84 cases of ACI patients admitted to our hospital from September 2017 to September 2019 and 90 cases of healthy subjects undergoing physical examination at the same time were selected as the research objects for prospective analysis. The concentrations of FSTL1, Bax and Bcl-2 in the peripheral blood of objects in the two groups were detected to analyze the diagnostic value of FSTL1, Bax and Bcl-2 for ACI, and the correlation of FSTL 1, Bax and Bcl-2 with the infarct size, treatment method and hemorrhagic transformation. Another 20 SD rats were purchased, among which 10 rats were randomly selected for ACI modeling. FSTL1 concentration, Bax and Bcl-2 protein expression in brain tissues of ACI rats and normal rats were detected. RESULTS: FSTL1 and Bax in peripheral blood of ACI patients were higher than those of healthy subjects (p<0.050), and Bcl-2 was lower than those of healthy subjects (p<0.050). It was detected that FSTL1, Bax and Bcl-2 had good diagnostic value for patients with ACI (p<0.001). FSTL1 and Bax decreased while Bcl-2 increased in patients treated with thrombolytic therapy (p<0.050). And FSTL1, Bax and Bcl-2 were closely related to infarct size and hemorrhagic transformation (p<0.050). Logistic regression analysis showed that NIHSS score, atrial fibrillation, infarct volume, FSTL1 and Bax were independent risk factors affecting hemorrhagic transformation in ACI patients (p<0.050), and Bcl-2 was an independent protective factor affecting hemorrhagic transformation in ACI patients (p<0.050). The concentration of FSTL1 and the expression of Bax protein in rat brain tissue were also higher than that in normal rats, while Bcl-2 was lower than that in normal rats (p < 0.001). CONCLUSIONS: FSTL1, Bax and Bcl-2 are involved in the occurrence and development of ACI and are closely related to the hemorrhagic transformation of patients. The mechanism by which FSTL1 promotes the occurrence of ACI might be related to promoting the occurrence of inflammatory responses in the brain tissue of patients or accelerating the apoptosis of neurons.

LncRNA RUNX1-IT1 inhibits proliferation and promotes apoptosis of hepatocellular carcinoma by regulating MAPK pathways.

OBJECTIVE: The aim of this study was to investigate the exact role of long non-coding RNA (lncRNA) RUNX1-IT1 in regulating the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells, and to explore the possible underlying mechanism. PATIENTS AND METHODS: LncRNA RUNX1-IT1 expressions in paired HCC tissues (cancer and paired non-cancer tissues) (n=80) and HCC cell lines were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The effects of lncRNA RUNX1-IT1 on the proliferation and apoptosis of HCC cells were estimated by cell counting kit-8 (CCK-8) and tunnel assay, respectively. Furthermore, the association between lncRNA RUNX1-IT1 expression and the overall survival of patients was analyzed by the survival curve. RESULTS: The expression level of lncRNA RUNX1-IT1 significantly decreased in HCC tissues. The overexpression of lncRNA RUNX1-IT1 remarkably inhibited the proliferation and induced the apoptosis of HCC cells. On the contrary, the knockdown of lncRNA RUNX1-IT1 remarkably enhanced the ability of proliferation and repressed the apoptosis of the HCC cells. In addition, lower expression of lncRNA RUNX1-IT1 indicated the worse prognosis of HCC patients. CONCLUSIONS: We found that lncRNA RUNX1-IT1 played an important role in the development of HCC by participating in cell proliferation and apoptosis. Thus, lncRNA RUNX1-IT1 might be a powerful candidate as a prognostic biomarker and therapeutic target for HCC.

Azithromycin influences airway remodeling in asthma via the PI3K/Akt/MTOR/HIF-1α/VEGF pathway.

Asthma is a respiratory disease that affects people of all walks of life, and is a hotspot of continuous research, with significant manpower and resources invested in its study. Airway remodeling is an important associated pathological change, and a mark of the irreversible damage produced by asthma. It involves compositional and functional changes in the cells of the airway walls, leading to reversible structural changes, and complicating treatment. Airway remodeling is mediated by different inflammatory pathways which have been targeted for treatment, with good results. However, given its complexity, systematic study of the pathogenesis of airway remodeling is still needed, and additional targeted therapies are necessary. Macrolide drugs, such as erythromycin, azithromycin, and clarithromycin, have antibacterial effects and also influence the cytokine secretion of macrophages and T-lymphocytes. They have direct effects on a variety of cytokines, inhibiting inflammation and reducing airway reactivity. In this study, we investigated the protective effect of azithromycin on airway remodeling through the phosphoinositol-3 kinase/Akt/mechanistic target of rapamycin kinase/hypoxia-inducible factor 1α (HIF-1α)/vascular endothelial growth factor (VEGF) pathway. We observed that a long course of azithromycin could significantly reduce airway reactivity and ovalbulmin-induced pathological alterations in asthmatic mice. Gene expression analysis confirmed that HIF-1α and VEGF were significantly down-regulated following a long course of azithromycin administration.

Evidence that the biofungicide Serenade (Bacillus subtilis) suppresses clubroot on canola via antibiosis and induced host resistance.

This study investigated how the timing of application of the biofungicide Serenade (Bacillus subtilis QST713) or it components (product filtrate and bacterial cell suspension) influenced infection of canola by Plasmodiophora brassicae under controlled conditions. The biofungicide and its components were applied as a soil drench at 5% concentration (vol/vol or equivalent CFU) to a planting mix infested with P. brassicae at seeding or at transplanting 7 or 14 days after seeding (DAS) to target primary and secondary zoospores of P. brassicae. Quantitative polymerase chain reaction (qPCR) was used to assess root colonization by B. subtilis as well as P. brassicae. The biofungicide was consistently more effective than the individual components in reducing infection by P. brassicae. Two applications were more effective than one, with the biofungicide suppressing infection completely and the individual components reducing clubroot severity by 62 to 83%. The biofungicide also reduced genomic DNA of P. brassicae in canola roots by 26 to 99% at 7 and 14 DAS, and the qPCR results were strongly correlated with root hair infection (%) assessed at the same time (r = 0.84 to 0.95). qPCR was also used to quantify the transcript activity of nine host-defense-related genes in inoculated plants treated with Serenade at 14 DAS for potential induced resistance. Genes encoding the jasmonic acid (BnOPR2), ethylene (BnACO), and phenylpropanoid (BnOPCL and BnCCR) pathways were upregulated by 2.2- to 23-fold in plants treated with the biofungicide relative to control plants. This induced defense response was translocated to the foliage (determined based on the inhibition of infection by Leptosphaeria maculans). It is possible that antibiosis and induced resistance are involved in clubroot suppression by Serenade. Activity against the infection from both primary and secondary zoospores of P. brassicae may be required for maximum efficacy against clubroot.

Dreaming during sevoflurane or propofol short-term sedation: a randomised controlled trial.

Prior reports suggest that dreaming during anaesthesia is dependent on recovery time. Dreaming during sedation may impact patient satisfaction. The current study explores the incidence and content of dreaming during short-term sedation with sevoflurane or propofol and investigates whether dreaming is affected by recovery time. A total of 200 women undergoing first trimester abortion (American Society of Anesthesiologists physical status I) participated in the study. Patients were randomly assigned to receive either sevoflurane or propofol for short-term sedation. Patients were interviewed upon emergence with the modified Brice questionnaire. The results showed the incidence of dreaming was significantly different between anaesthesia groups with 60% (60/100) of the sevoflurane group and 33% (33/100) of the propofol group (P=0.000). However, recovery time did not significantly differ between groups. In the sevoflurane group, a greater number of dreamers could not recall what they had dreamed about (P=0.02) and more patients reported dreams that had no sound (P=0.03) or movement (P=0.001) compared with dreamers in the propofol group. Most participants reported dreams with positive emotional content and this did not significantly differ between groups. Anaesthesia administered had no effect on patient satisfaction. The results suggest that the incidence of dreaming was not affected by recovery time. Patient satisfaction was not influenced by choice of sedative and/or by the occurrence of dreaming during sevoflurane or propofol short-term sedation.

The valence strength of negative stimuli modulates visual novelty processing: electrophysiological evidence from an event-related potential study.

In natural settings, the occurrence of unpredictable infrequent events is often associated with emotional reactions in the brain. Previous research suggested a special sensitivity of the brain to valence differences in emotionally negative stimuli. Thus, the present study hypothesizes that valence changes in infrequent negative stimuli would have differential effects on visual novelty processing. Event-related potentials (ERPs) were recorded for highly negative (HN), moderately negative (MN) and Neutral infrequent stimuli, and for the frequent standard stimulus while subjects performed a frequent/infrequent categorization task, irrespective of the emotional valence of the infrequent stimuli. The infrequent-frequent difference waves, which index visual novelty processing, displayed larger N2 amplitudes during HN condition than during MN condition which, in turn, elicited greater N2 amplitude than the Neutral condition. Similarly, in the infrequent-frequent difference waves, the frontocentral P3a and parietal LPC (late positive complex) elicited by the HN condition were more negative than those by MN stimuli, which elicited more negative amplitudes than the Neutral condition. This suggests that negative emotions of diverse strength, as induced by negative stimuli of varying valences, are clearly different in their impact on visual novelty processing. Novel stimuli of increased negativity elicited more attentional resources during the early novelty detection, and recruited increased inhibitive and evaluative processing during the later stages of response decision and reaction readiness, relative to novel stimuli of reduced negativity.

Identification of a novel HLA-A allele HLA-A*2451 in a Chinese donor.

A novel human leucocyte antigen-A (HLA-A) allele, A*2451, has been identified during routine sequence-specific oligonucleotide typing and sequence-based typing of a sample from a registered donor of the Chinese Marrow Donor Program. The A*2451 allele differs from the closest matching allele A*2415 by one nucleotide substitution in exon 3, nt 363 G-->A, resulting in an amino acid change from M ATG to I ATA at codon 121.

VEGF, bFGF and their receptors at the fetal-maternal interface of the rhesus monkey.

Placental development involves trophoblast outgrowth and a coordinated angiogenesis in the implantation site. In this study, expression of angiogenic factors, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), VEGF receptors, kinase insert domain-containing region (KDR), and bFGF receptor Flg was characterized at the maternal-embryonic boundary of the rhesus monkey on Day 17, 19, 28 and 34 of pregnancy. Immunohistochemistry and in situ hybridization showed that VEGF mRNA and protein were both strongly expressed in the cytotrophoblast, the blood vessels and certain immunocytes. These sites were also immunopositive for KDR. In addition to the vascular endothelial cells and the vascular smooth muscle cells, the protein and mRNA for bFGF were also detected in cyto/syncytiotrophoblast bilayer, whereas the staining for Flg protein was mainly localized in the cytotrophoblast cells. The staining degree of VEGF and bFGF in the villi gradually decreased with the development of placenta. Strong expression of bFGF, Flg and KDR was also detected in the decidual cells. These data suggest that VEGF and bFGF may be involved in angiogenesis, cytotrophoblast proliferation and migration during early stage of placentation in the rhesus monkey.