Objective : To explore the role of semaphoring 6d(SEMA6D) in the malignant progression of triple-negative breast cancer(TNBC). Methods : Bioinformatics and Immunohistochemistry(IHC) were used to analyze the expression level of SEMA6D in TNBC and paracancer non-tumor tissues and its relationship with patients′ clinicopathological features. MDA-MB-231 cell line stably knocking down the expression of SEMA6D was constructed, and the effects of SEMA6D on migration and invasion of TNBC cells were investigated by Wound-healing assays and Transwell assays. cBioPortal and GEPIA2 databases were used to screen out the gene negatively associated with it, namely aurora kinase A(AURKA). Bioinformatics and IHC were used to analyze the expression level of AURKA in TNBC and paracancer non-tumor tissues and its relationship with patients' clinicopathological features. Western blot assay was used to analyze the expression of AURKA and the effect of epithelial-mesenchymal transition(EMT) makers Claudin-1, N-cadherin and Vimentin after knocking downSEMA6D. Results: Bioinformatics analysis and IHC results showed that the expression of SEMA6D in TNBC tissues was significantly lower than that in paracancer non-tumor tissues(bothP<0.05). The expression of AURKA in TNBC tissues was significantly higher than that in paracancer non-tumor tissues(bothP<0.05), SEMA6D and AURKA were significantly negatively correlated in TNBC(P<0.01). Both low expression of SEMA6D and high expression of AURKA were positively correlated with tumor size, tumor histological grade, clinical stage and lymph node metastasis in TNBC patients(allP<0.05). The knockdown ofSEMA6Dsignificantly promoted the migration and invasion ability of TNBC cells(bothP<0.01). Western blot results showed that the knockdown ofSEMA6Dupregulated AURKA expression, promoted the expression of N-cadherin and Vimentin, and inhibited the expression of Claudin-1 in tumor cells. Conclusion Down-regulation of SEMA6D expression in TNBC may be involved in the malignant progression of TNBC through up-regulation of AURKA expression and promotion of EMT.

Objective : To investigate the expression of Keratin 14(KRT14) in Basal-like Breast Cancer(BLBC) and its biological functions and mechanisms. Methods : The expression levels of KRT14 mRNA in BLBC and para-cancer breast tissues were analyzed using The Cancer Genome Atlas(TCGA) database. qPCR, Western blot(WB), and immunohistochemistry were employed to detect KRT14 expression in BLBC and adjacent normal tissues, and its correlation with clinicopathological features was analyzed. KRT14 overexpression and knockdown were performed in breast cancer cells, and cell scratch and transwell assays were performed to evaluate changes in migration and invasion abilities. To investigate the expression of proteins related to the Wnt/β-catenin signaling pathway, including catenin Beta 1(β-catenin), wingless-type MMTV integration site family, member 1(Wnt1), matrix metallopeptidase 7(MMP7), and cellular myelocytomatosis viral oncogene homolog(c-Myc), as well as the cellular localization of β-catenin, WB and immunofluorescence(IF) techniques were employed. Additionally, a Wnt/β-catenin signaling pathway inhibitor was used to verify the mechanism of action of KRT14. Results : The expression of KRT14 was significantly higher in BLBC tissues compared to normal tissues(P<0.05), and was associated with higher T stage and histological grade(P<0.05). The overexpression of KRT14 significantly enhanced the migration and invasion abilities of breast cancer cells, while the knockdown of KRT14 significantly reduced those abilities(P<0.01). The overexpression of KRT14 can increase the expression levels of Wnt/β-catenin pathway-related proteins β-catenin, Wnt1, MMP7, and c-Myc, thereby activating the Wnt/β-catenin pathway. Moreover, the inhibition of this pathway can eliminate the effects of KRT14 on cell migration and invasion. Conclusion The high expression of KRT14 in BLBC may promote the migration and invasion of breast cancer cells through the Wnt/β-catenin signaling pathway.

Objective : To investigate the expression of(heat shock protein a member 8,HSPA8) in breast cancer and its effect on tumor biological behaviors. Methods: Bioinformatics analysis and immunohistochemistry assays were used to detect the expression of HSPA8 in breast cancer and adjacent non-tumor breast tissues,and the relationship between its expression and clinicopathological characteristics of breast cancer patients was analyzed.Correlation between HSPA8 expression and prognosis of breast cancer patients was analyzed by Kaplan-Meier Plotter database.HSPA8 knockdown and over expression breast cancer stabilized cells were constructed,respectively.CCK-8,clone formation,Transwell,cell scratch,Western blot and immunofluorescence assay were used to detect the effects of HSPA8 on the proliferation,invasion and migration of breast cancer cells,and its effect on epithelial-mesenchymal transition(EMT). Results : Bioinformatics analysis and immunohistochemistry assay revealed that the expression of HSPA8 in breast cancer tissues was higher than that in adjacent non-tumour tissues(P<0.05),and its expression level of the protein was significantly and positively correlated with the tumor size,histological grade,lymph node metastasis and Ki-67 proliferation index(P<0.05).The Kaplan-Meier survival curve showed that high expression of HSPA8 was significantly associated with poor prognosis in breast cancer patients(P<0.05).CCK-8,clone formation,transwell,cell scratch,Western blot and immunofluorescence assay showed that knockdown of HSPA8 expression could significantly inhibit the proliferation,invasion,migration function and EMT of breast cancer cells(P<0.05),while overexpression of HSPA8 could significantly promote the proliferation,invasion,migration function and EMT of breast cancer cells(P<0.05). Conclusion HSPA8 is highly expressed in breast cancer tissues,which is closely related to disease progression and the malignant phenotype of breast cancer,suggesting that HSPA8 may be a potential biological target for breast cancer treatment.

Objective: To explore the function of cluster needling at scalp points therapy on regulating differential protein's expression at different time points in middle cerebral artery occlusion (MCAO) model rats. Methods: Fifty-four rats were divided into three groups randomly and 18 rats in each group. The groups respectively were the model group (group M, n = 18), cluster needling at scalp points group (group C, n = 18), false operation group (group F, n = 18). Each group was then assigned in three subgroups, including 24-h, 7-day, and 14-day subgroups. Six rats in each subgroup. Acupuncture at Baihui (GV20) and 2 points beside Baihui, which was 3-4 mm away from the midline. Longa score was used to eval-uated neurological effects. Proteomics methods were used to identify differentially expression proteins with a standard of fold change greater than 1.5 and P<.05 at different times. Results: 1. Nerve function scoring: The nerve function scores at 7 and 14 days decreased in group C, which showed better neural function than group M (P<.05). 2. Fold change in proteins:Group M showed 932 differentially expressed proteins compared with group F, and among them, 414 proteins showed significant changes in expression after acupuncture. The expression levels of Cdc42 and GFAP were increased, and Mag, Shank2, and MBP levels were decreased. In the Gene Ontology analysis, the cellular component consisted of the terms cytoplasm, cytoskeleton, lysosome, and plasma membrane. The main related biological processes were cell-cell signaling, protein transport, aging, and cell adhesion. Many synaptic and metabolic pathways were found by KEGG analysis. Conclusion: Cluster needling at scalp acupoints can improve the nerve function score and improve dyskinesia in MCAO model rats. Cluster needling at scalp acupoints can regulate the expression of 414 proteins, including Cdc42, GFAP, Mag, Shank2, and MBP, which are related to cerebral ischemia. The differential proteins are major concentration in cytoplasm, cytoskeleton, lysosomes, and plasma mem-brane, participate in cell-cell signaling, protein transport, aging, and cell adhesion, and act through multiple synaptic and metabolic pathways to exert their biological functions.

DNA methylation is a significant epigenetic modification mode, which plays an important role in embryo reprogramming, stem cell differentiation and tumor occurrence. The ten-eleven translocation (TET) enzyme is a crucial demethylation enzyme, which can catalyze 5-methylcytosine(5mC) to 5-hydroxymethylcytosine(5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine(5caC). These bases represent the epigenetic modifications of DNA and regulate the process of DNA methylation. Understanding the role of TET enzyme in regulating the DNA methylation modification and gene expression can help us to gain the knowledge for the normal growth development and epigenetic regulation in human diseases.
5-Methylcytosine ; metabolism ; Cell Differentiation ; DNA ; DNA Methylation ; DNA-Binding Proteins ; Epigenesis, Genetic ; Humans

Objective To evaluate the applicability of the Chinese version of SF-36 ( v. 2 ) scale for evaluating the quality of life of hospitalized patients with chronic heart failure. Methods From September 2013 to December 2014, 159 patients with chronic heart failure(NYHA I-IV), who were older than 18 years, clear mind and well self-expressed, were selected as participants. Questionnaire surveys included general survey and SF-36(v. 2) scale. Internal consistency reliability, binary reliability and construct validity were all analyzed as indicators to evaluate SF-36 ( v. 2 ) scale. Results A total of 159 questionnaires were issued and 159 valid questionnaires were recovered. The eight dimensions of SF-36(v. 2) scale including physical function (PF), role-physical (RP), bodily pain (BP), general health (GH), social function (SF), vitality (VT), role-emotion(RE), and mental health (MH) score conversion were (41.57 ±24.86), (48.35 ±21.64), (69.18 ± 25. 68), (31. 28 ± 16. 01), (48. 90 ± 19. 53), (45. 05 ± 22. 76), (59. 43 ± 24. 31), (57. 55 ± 19. 03); the floor effects were 2. 5%, 4. 4%, 3. 1%, 4. 4%, 3. 1%, 6. 3%, 0. 6%, 1. 3%; the ceiling effects were 0. 0%, 3. 8%, 21. 4%, 0. 0%, 0. 0%, 1. 9%, 3. 1%, 0. 0%. The item-convergent validity all achieved the standard (r≥0. 4), and the total scaling success rate of item-convergent was 100. 00%; the dimensions′success rates of item-discriminant validity of RP, BP, RE and SF were all 100%, the rest of four dimensions were PF 95. 71%, GH 85. 71%, VT 89. 29%, MH 94. 29%, and the total success rate was 94. 69%. Internal consistency reliability ranged from 0. 738 to 0. 919; the binary reliability ranged from 0. 808 to 0. 963. Within factors analysis, two common factors were confirmed, separately representing physical health and mental health, altogether making contribution of 61. 66% cumulative variance. Conclusions As the revision of SF-36(v. 1), SF-36(v. 2) scale seemed more friendly in layout for questions and answers, the floor and ceiling effects significantly reduced. Additionally, it also shows good reliability and validity in the evaluation of quality of life of hospitalized patients with chronic heart failure, and the SF-36(v. 2) scale can be used to evaluate the quality of life ( QOL) of patients with chronic heart failure.