Objective To verify the mechanism of bromodomain PHD finger transcription factor(BPTF)affecting develop-ment of glioma and ferroptosis by regulating the expression of solute carrier family 40 member 1(SLC40A1)in glioma cells by in v itro and invivo experiments.Methods U87MG cells were divided into sh-NC group,sh-BPTF group,sh-BPTF+ov-NC group and sh-BPTF+ov-SLC40A1 group.Cell lines were stably transfected by lentivirus,and the transfection efficiency was verified by qRT-PCR and Western blotting.Cell proliferation ability was detected by CCK-8 and plate clone formation test.Cell migra-tion and invasion ability were detected by Transwell.Cells were injected subcutaneously into nude mice to detect the tumor growth.To evaluate theferroptosis of cells,the reactive oxygen species(ROS)level,iron content,malondialdehyde(MDA)con-tent and reduced glutathione/oxidized glutathione(GSH/GSSG)ratio in cells were detected by corresponding kits.Co-immuno-precipitation(Co-IP)experiment was used to verify the interaction between BPTF and c-Myc protein.Chromatin immunoprecipi-tation(ChIP)experiment was used to verify that BPTF and c-Myc combined with SLC40A1 promoter.Double luciferase reporter gene experiment was used to verify theeffect of BPTF on SLC40A1 transcription.Results After BPTF knockdown,theexpres-sion of SLC40A1 in glioma cell was decreased,cell proliferation,migration,invasion in vitro,and tumor growth in vivo were in-hibited,iron content,ROS level and MDA content in cells were increased,and GSH/GSSG ratio in cell was de-creased.Overexpression of SLC40A1 reversed the inhibitory effect of BPTF on the proliferation,migration,invasion and tumor grow th in vivo,decreased the iron content,ROS level and MDA content in cells,and increased the GSH/GSSG ratio in cells.There was an interaction between BPTF and c-Myc proteins in gliomacells.A potential binding site of c-Mycin SLC40A1 promoter was verified,BPTF and c-Myc protein bound to SLC40A1 promoter.BPTF knockdown reduced the transcriptional ac-tivity of SLC40A1 promoter,and BPTF knockdown after binding site mutation did not affect the transcriptional activity of SLC40A1 promoter.Conclusion BPTF may upregulate the expression of its downstream target gene SLC40A1 by interacting with c-Myc,thereby inhibiting ferroptosis in glioma cells and promoting glioma progression.

Objective To verify the mechanism of bromodomain PHD finger transcription factor(BPTF)affecting develop-ment of glioma and ferroptosis by regulating the expression of solute carrier family 40 member 1(SLC40A1)in glioma cells by in v itro and invivo experiments.Methods U87MG cells were divided into sh-NC group,sh-BPTF group,sh-BPTF+ov-NC group and sh-BPTF+ov-SLC40A1 group.Cell lines were stably transfected by lentivirus,and the transfection efficiency was verified by qRT-PCR and Western blotting.Cell proliferation ability was detected by CCK-8 and plate clone formation test.Cell migra-tion and invasion ability were detected by Transwell.Cells were injected subcutaneously into nude mice to detect the tumor growth.To evaluate theferroptosis of cells,the reactive oxygen species(ROS)level,iron content,malondialdehyde(MDA)con-tent and reduced glutathione/oxidized glutathione(GSH/GSSG)ratio in cells were detected by corresponding kits.Co-immuno-precipitation(Co-IP)experiment was used to verify the interaction between BPTF and c-Myc protein.Chromatin immunoprecipi-tation(ChIP)experiment was used to verify that BPTF and c-Myc combined with SLC40A1 promoter.Double luciferase reporter gene experiment was used to verify theeffect of BPTF on SLC40A1 transcription.Results After BPTF knockdown,theexpres-sion of SLC40A1 in glioma cell was decreased,cell proliferation,migration,invasion in vitro,and tumor growth in vivo were in-hibited,iron content,ROS level and MDA content in cells were increased,and GSH/GSSG ratio in cell was de-creased.Overexpression of SLC40A1 reversed the inhibitory effect of BPTF on the proliferation,migration,invasion and tumor grow th in vivo,decreased the iron content,ROS level and MDA content in cells,and increased the GSH/GSSG ratio in cells.There was an interaction between BPTF and c-Myc proteins in gliomacells.A potential binding site of c-Mycin SLC40A1 promoter was verified,BPTF and c-Myc protein bound to SLC40A1 promoter.BPTF knockdown reduced the transcriptional ac-tivity of SLC40A1 promoter,and BPTF knockdown after binding site mutation did not affect the transcriptional activity of SLC40A1 promoter.Conclusion BPTF may upregulate the expression of its downstream target gene SLC40A1 by interacting with c-Myc,thereby inhibiting ferroptosis in glioma cells and promoting glioma progression.

OBJECTIVES: The high morbidity and mortality of colorectal cancer (CRC) have posed great threats to human health. Circular RNA (circRNA) and microRNA (miRNA), acting as competing endogenous RNAs (ceRNAs), have been found to play vital roles in carcinogenesis. This paper aims to construct a circRNA/miRNA/mRNA regulatory network so as to explore the molecular mechanism of CRC. METHODS: The sequencing data of circRNA from CRC were obtained from Gene Expression Omnibus (GEO). The differential circRNA was screened and its structure was identified by Cancer-specific CircRNA Database (CSCD); the sequencing data of miRNA and messenger RNA (mRNAs) were downloaded from The Cancer Genome Atlas (TCGA) database and the differentially expressed genes were screened; the corresponding miRNA of differential circRNAs were predicted by CircInteractome database; DIANA, Miranda, PicTar, and TargetScan databases were used to predict the target genes of different miRNAs; the target genes from Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were enriched by R language; String database combined with Cytoscape 3.7.2 software was used to construct protein-protein interaction (PPI) network and hub genes were screened; the expressions of mRNAs in the Top10 hub genes were verified in CRC. The network diagrams of circRNAs/miRNAs/mRNAs and circRNAs/miRNAs/Top10 hub mRNAs were constructed by Cytoscape3.7.2. Real-time PCR was used to examine the expression levels of hsa_circRNA_0065173, hsa-mir-450b, hsa-mir-582, adenylate cyclase 5 (ADCY5), muscarinic acetylcholine receptor M2 (CHRM2), cannabinoid receptor 1 (CNR1), and lysophosphatidic acid receptor 1 (LPAR1) in the CRC tissues and the adjacent normal tissues. RESULTS: A total of 14 differential circRNAs were identified, and 8 were found in CSCD; 34 miRNAs targeted by circRNAs were obtained. The PPI network was constructed, and the Top10 hub genes were identified, which were CHRM2, melanin concentrating hormone receptor 2 (MCHR2), G-protein gamma 3 subunit (GNG3), neuropeptide Y receptor Y1 (NPY1R), CNR1, LPAR1, ADCY5, adenylate cyclase 2 (ADCY2), gamma 7 (GNG7) and chemokine 12 (CXCL12), respectively. The expressions of Top 10 hub genes were also verified, and the results showed that the Top 10 hub genes were down-regulated in CRC; the constructed network diagram showed that hsa_circRNA_0065173 may regulate ADCY5, CHRM2, and Hsa-mir-450b by modulating hsa-mir-450b and hsa-mir-582. CNR1 and LPAR1 genes might serve as potentially relevant targets for the treatment of CRC. Real-time PCR results showed that the expression levels of hsa_circRNA_0065173, ADCY5, CHRM2, CNR1 and LPAR1 in the CRC tissues were significantly reduced compared with the adjacent normal tissues (all P<0.05); the expression levels of hsa-mir-450b and hsa-miR-582 were significantly increased (both P<0.05). CONCLUSIONS In this study, a potential circRNAs/miRNAs/mRNAs network is successfully constructed, which provides a new insight for CRC development mechanism through ceRNA mediated by circRNAs.
Colorectal Neoplasms/genetics* ; Computational Biology/methods* ; Gene Regulatory Networks ; Humans ; MicroRNAs/genetics* ; RNA, Circular/genetics* ; RNA, Messenger/genetics*

OBJECTIVE: To assess the application value of mapping allele with resolved carrier status (MaReCs) technique for preimplantation genetic testing (PGT). METHODS: The characteristics of MaReCs for PGT and outcome of patients were retrospectively analyzed. RESULTS: Compared with those who could not use the technique, carriers who have used the MaReCs technique were younger, had significantly higher level of anti-Mullerian hormone, more antral follicles, occytes, mature occytes, biopsied embryos and euploid embryos, and lower risks for de novo chromosomal abnormality (P<0.05). It was necessary for couples with fewer oocytes, mature oocytes and balstocyst to preserve discarded embryos to facilitate the test. Carriers who have used the MaReCs technique had higher clinical pregnancy rate and abortion rate compared with those undergoing routine PGT, albeit no significant difference was found between the two groups (P> 0.05). Carriers undergoing MaReCs test could preferentially select embryos with normal chromosome structures for the transfer. CONCLUSION Application of MaReCs has a prerequisite for having a minimum number of occytes and biopsied embryos and using discarded embryos sometimes. MaReCs is efficient for the detection of carrier status of embryos and attaining higher rate of pregnancy and live birth, which can significantly improve the outcome for couples carrying chromosomal translocations.
Alleles ; Aneuploidy ; Blastocyst ; Female ; Fertilization in Vitro ; Genetic Testing ; Humans ; Pregnancy ; Preimplantation Diagnosis ; Retrospective Studies ; Translocation, Genetic

Objective To investigate the role of PKCι, YAP1 and high-risk HPV infection in the local immune microenvironment of cervical cancer. Methods We chose 80 cases of normal tissue of the cervix (NCT), cervical low-grade squamous intraepithelial lesion (LSIL), cervical high-grade squamous intraepithelial lesion (HSIL) and early cervical squamous cell carcinoma (SCC) each. Four groups were collected.The infection rate of high-risk HPV in four groups was determined by real-time fluorescence PCR method. The expression levels of PKCι, YAP1, CD4 and CD8 in four groups were measured and correlated by IHC and clinicopathologic features were also analyzed. Results The differences of high-risk HPV infection rate and PKCι, YAP1, CD4, CD8 positive rate among groups of NCT, LSIL, HSIL and SCC had statistical significance (P < 0.05). The level of cervical lesions was positively associated with high-risk HPV infection and positive PKCι, YAP1, CD8 expression (P < 0.05), while negatively associated with positive CD4 expression (P < 0.05). HPV infection and positive PKCι, YAP1, CD8 expression were positively correlated with each other in SCC, while were all negatively correlated with positive CD4 expression(P < 0.05). The differences of HPV infection, PKCι, YAP1 and CD8 positive expression were significant in different levels of differentiation and vascular invasion of SCC (P < 0.05). Conclusion The patients with cervical lesions are often accompanied by high-risk HPV infection and abnormal expression of PKCι, YAP1, CD4 and CD8, which may have synergistic effects on each other, causing the local immunosuppression microenvironment of SCC. It provides a possible strategy for the study of pathogenesis, diagnosis and treatment of cervical cancer.

Objective:To explore the clinical significance of classification and grading of ovarian serous tumors (OST) and the correlation of classification and grading with expressions of p53 and Ki-67 proteins, so as to provide a basis for accurate diagnosis and reasonable treatment of OST.Methods:A total of 100 paraffin-embedded ovarian tissues were collected from Haikou Affiliated Hospital of Central South University Xiangya School of Medicine from January 2012 to November 2017. The classification and grading of OST was based on the World Health Organization (WHO) ovarian tumor tissue classification, the work conference on borderline ovarian tumors which was held in August 2013 in Maryland, United States, and the two-level tissue classification system of the United States MD Anderson Cancer Center. The 100 cases of ovarian tissues included 10 cases of normal ovarian tissues (NOT), 12 cases of ovarian serous cystadenoma (OSA), 18 cases of ovarian serous borderline tumor (OSBT), 22 cases of low-grade serous carcinoma (LGSC), and 38 cases of high-grade serous carcinoma (HGSC). The expressions of p53 and Ki-67 proteins in paraffin-embedded tissues were detected by immunohistochemistry. The clinical characteristics of patients with different types of OST and their relationships with the expression of p53 protein and Ki-67 positive index were analyzed.Results:Among OSBT, LGSC and HGSC groups, the proportion of patients with onset age > 50 years old [38.9% (7/18), 45.5% (10/22), 73.7% (28/38)], poor differentiation [0 (0/18), 0 (0/22), 100.0% (38/38)], stage Ⅲ-Ⅳ [5.6% (1/18), 27.3% (6/22), 39.5% (15/38)] increased sequentially, and the differences were statistically significant (all P < 0.05). Among NOT, OSA, OSBT, LGSC and HGSC groups, there were significant differences in the positive rate of p53 protein [20.0% (2/10), 25.0% (3/12), 27.8% (5/18), 31.8% (7/22), 57.9% (22/38)] and Ki-67 positive index [(10.40±0.00)%, (31.49±6.53)%, (42.81±6.84)%, (74.29±6.54)%, (77.04±8.88)%] (all P < 0.05). In the patients with ovarian serous carcinoma (LGSC + HGSC), there was no significant difference in the positive rate of p53 protein and Ki-67 positive index between the onset age ≤ 50 years old and > 50 years old groups (both P > 0.05), but there were significant differences between the patients with different tissue differentiation grade, clinical stage and metastasis (all P < 0.05). There was significant difference in Ki-67 positive index among NOT, OSA, OSBT, LGSC and HGSC patients with positive expression of p53 protein ( P < 0.01). Conclusions:The classification and grading of OST is related to onset age, tissue differentiation and clinical stage, which can be used to guide treatment and judge prognosis. The positive rate of p53 protein and Ki-67 positive index increase with advancing grade of breast lesions, which may be related to the occurrence and development of OST.

Objective:To investigate the prognostic value of systemic inflammatory score (SIS) in prognostic assessment of patients with unresectable metastatic colorectal cancer (mCRC).Methods:The clinical data of 130 patients with unresectable mCRC in Affiliated Haikou Hospital of Xiangya Medical College of Central South University from January 2014 to December 2016 were retrospectively analyzed. The relationship between SIS and clinicopathological characteristics of unresectable mCRC patients was also analyzed. The survival analysis was made by using Kaplan-Meier. The risk factors affecting the prognosis of unresectable mCRC patients were analyzed by using Cox regression model to make univariate and multivariate analysis.Results:According to SIS results, patients were divided into 0-score group (40 cases), 1-score group (58 cases), and 2-score group (32 cases). There were no significant differences in different SIS constitution patients stratified by age, gender, primary tumor location, functional status score, tissue type, RAS gene status, number of metastatic organs, peritoneal spread and molecular targeted therapy (all P > 0.05). Kaplan-Meier survival analysis showed that the 5-year overall survival rates of SIS 0-score, 1-score and 2-score group were 37.5%, 19.0%, 6.3%, respectively; and the difference was statistically significant ( χ2 = 3.152, P<0.01). Cox regression survival analysis showed that female, primary tumor location in right side and SIS (scores of 1-2) were independent risk factors for overall survival in patients with unresectable mCRC (all P<0.05). Conclusion:SIS may be an important indicator for prognostic assessment of patients with unresectable mCRC, and patients with high SIS have poor prognosis.

Objective:To deeply understand needs and problems of on-site nursing service training for "network nurses", so as to provide a reference for the construction of "Internet + Nursing Service" training content system standards.Methods:Using the purposive sampling method and based on principle of maximum differentiation, a total of 12 "network nurses" in Ningbo were selected from March to May in 2020 to conduct semi-structured interviews. Braun thematic analysis method was used to analyze data and extract themes.Results:A total of 4 themes were obtained. The first was that the requirements of practicing safety education and training included risk prevention training, adverse event handling process training, emergency response plans and complication handling training. The second was the needs of humanistic care education and training, including on-site service language, code of conduct training, nurse-patient communication training and health education knowledge training. The third was the demand for diversified training forms. Fourthly, factors affecting training quality.Conclusions:"Network nurses" have a greater demand for occupational safety education and humanistic care education. It is recommended that multiple parties should cooperate to improve the "Internet +Nursing Service" emergency laws and regulations as soon as possible, provide diversified training models, reserve high-quality "Internet + Nursing Service" talents and promote continuous and standardized development of "Internet + Nursing Service".

Objective To study the specific killing effect in human carcinoma cells aftercombination treatment of radiation and p53 gene regulated by a radiation-enhanced promoter.Methods Aplasmid pE6 (TATA)-p53 was constructed.After irradiation,the expression of P53 was detected withWestern blot assay,apoptosis was detected by Annexin V-FITC,and cell survival was detected byclonogenic assay then the sensitivity enhancement ratio (SER) was analyzed for HeLa and A549 cells.Results The expression of P53 were increased in the irradiated cells and 6 Gy irradiation triggered thestrongest activity.After p53 transfection,radiation-induced apoptosis was obviously enhanced incomparison with the control group without gene transfection (F =11.018,10.736,P ＜ 0.05).The SER ofp53-promoter was 2.36 for A549 cells and 2.56 for Hela cells.Conclusions The p53-plasmid promotercould induce apoptosis and enhance the radiosensitivity of tumor cells,which may provide a noveltherapeutic strategy for cancer treatment.

Objective To investigate the effects of altered deceleration capacity of heart rate (DC) and heart rate deceleration runs(DRs) on the short-term prognosis of patients with acute myocardial infarction (AMI) .Methods 115 patients confirmed with AMI within 7 days as AMI group ,50 persons without MI were selected as control group .The deceleration capacity of heart rate and heart rate deceleration runs were detected by the 24 hours Holter within 7 to 14 after onset of AMI ,all the patients were divided in-to high risk group ,medium risk group and low risk group through the Holter results .Left ventricular ejection fraction (LVEF) was evaluated by echocardiography .Compare the risk of the AMI group with the control group .Meanwhile the AMI patients were fol-lowed up for mean(9 .2 ± 1 .7)months .Observe the presence of major adverse cardiovascular events (MACE) .All AMI patients were classified into MACE and non-MACE groups according to the presence of MACE .Analysis the risk factors of MACE .Results AMI group had a significant increase risk but decrease in deceleration capacity of heart rate (P<0 .05) as compared with the control group .In MACE group ,the rate of high risk was significant increasing(P=0 .005) ,but the rate of low risk was significant decrea-sing(P=0 .039) .The Logistic regression analysis showed that LVEF ,high risk group ,age and percutaneous transluminal coronary intervention(PCI) were independent risk factors on presence of MACE .Correlation r was 0 .33 ,0 .30 ,0 .23 ,0 .18 respectively ,and odds ratio was 4 .83 ,4 .28 ,2 .22 ,1 .58 respectively ,the correlation with deceleration capacity of heart rate was a little weaker than LVEF .The coefficient of determination r2 was 0 .528 when the LVEF and ages went into regression ,whereas that was 0 .635 when the LVEF ,high risk group and age went into regression together .Conclusion Capacity of heart rate decelerate was significantly weakened in AMI patients .LVEF ,the significantly decreased deceleration capacity of heart rate ,age and PCI were independent risk factors on presence of short-term MACE in AMI patients .The prognostic value of the DC and DRs was a little weaker than the LVEF .The better predictive value was obtained if the LVEF ,the significantly decreased deceleration capacity of heart rate ,age were combined .