Abstract:Objective：Accumulation of reactive oxygen species（ROS） in skeletal muscle cells promotes apoptosis，which represents a key mechanism underlying the loss of muscle mass during aging. Previously we have demonstrated that the transcription factor E2F1 promotes the transcription of the pro-apoptotic gene PERP in skeletal muscle cells（C2C12）. In the present stuy we investigated the epigenetic mechanism whereby E2F1 expression is stimulated by ROS in C2C12 cells. Methods：Real-time qPCR and Western blotting were used to examine gene expression levels. Chromatin immunoprecipitation was used to determine the interaction between protein and DNA. Results：We report that E2F1 levels were up-regulated in C2C12 exposed to H2O2. Up-regulation of E2F1 expression by H2O2 treatment was accompanied by erasure of dimethyl H3K9 from the E2F1 promoter. Further study revealed that H2O2 treatment promoted the binding of KDM3A，a histone demethylase，on the E2F1 promoter. In contrast，KDM3A depletion by siRNA significantly dampened the induction of E2F1 expression by H2O2 treatment. Concomitantly，KDM3A knockdown decreased the levels of acetylated H3 and trimethylated H3K4 but restored the levels of dimethylated H3K9 on the E2F1 promoter. Conclusion：Our data suggest that KDM3A may participate in H2O2 treatment-induced E2F1 trans-activation by modulating histone modifications on the E2F1 promoter.

Abstract:Objective：To optimize，screen and identify the bispecific c-Met/PD-L1 scFv fusion protein and to detect whether the different combinations of heavy chains and light chains can influence the biological activity of bispecific scFv fragments. Methods：Bioinformatics analysis，technique of gene engineered antibodies were introduced to design，optimize and construct bi-specific scFv fusion proteins. BLItz was used to analyze the affinity of those bi-specific scFv fusion proteins to c-Met and PD-L1，ELISA assay was used to detect their specific binding ability. Results：Bispecific scFv fusion proteins were produced successfully，BLItz and ELISA detection confirmed that the bispecific scFv fusion protein CP1 has higher affinity and specific binding ability. Conclusion：Different combinations of heavy and light chains can affect the affinity and specific binding activity of c-Met/PD-L1 scFv-Fc fusion proteins，CP1 has the strongest affinity and specificity for binding to c-Met and PD-L1 protein，it can be applied to the construction of c-Met/PD-L1 expression vector and subsequent research of c-Met/PD-L1 CAR-T cells.

Abstract:Objective：To study the relationship between mitochondrial augmenter of liver regeneration（ALR）and mitochondrial dysfunction in cell model of hepatocyte lipotoxicity. Methods：The 10% fatty acid-free bull serum albumin treated L-O2 cells were used as the control group（BSA group），and L-O2 cells treated with 0.2 mmol/L palmitic acid（PA）as a cell model of hepatocyte lipotoxicity（PA group）. Meanwhile，the ALR overexpression group（ALR-OE group）and the empty vector transfected control cell group（Vector group）were constructed and treated with BSA or PA，respectively. cell viability，lactate dehydrogenase（LDH）release，mitochondrial membrane potential changes，apoptosis and related protein expression levels were detected subsequently. Results：Compared with the BSA group，the intracellular lipid droplet increased，the cell viability decreased by about 50%，and the LDH release increased by 13 times，the expression of ALR in the mitochondria was significantly decreased in the PA group. There were no significant differences between the Vector group and the ALR-OE group under BSA treatment conditions. However，the cell viability increased by about 16%，LDH release is reduced by about 40%，mitochondrial membrane potential is increased by about 18%，cytochrome C release is reduced，and apoptosis decreases in the ALR-OE group compared with the Vector group under PA stimulation. Conclusion：ALR is involved in the onset of hepatocellular lipotoxicity，and targeting the regulation of mitochondrial ALR can inhibit the onset of lipotoxicity to a certain extent.

Abstract:Objective：To investigate the effect of geniposide on activity and endocytosis of murine macrophage RAW264.7 cells. Methods：The RAW264.7 cells were added with geniposide at different concentrations and further cultured for several hours. The proliferation of RAW264.7 cells was measured by CCK-8 assay. According to the results of CCK-8 assay，geniposide at appropriate concentrations were selected to culture with RAW264.7 cells for 24 h，and subsequently detected the pinocytosis activity of neutral red solution. After cultured with geniposide at the optimal concentration（2 mg/mL），RAW264.7 cells were detected the phagocytosis activity of Pseudomonas aeruginosa and intracellular bacterial survival. Results：Geniposide showed little cytotoxicity on RAW264.7 cells. The proliferation of RAW264.7 cells was not seriously affected by 0~8 mg/mL geniposide. Geniposide at 2 mg/mL could significantly promote the pinocytosis of neutral red solution by RAW264.7 cells，but inhibited the phagocytosis of Pseudomonas aeruginosa and intracellular bacterial survival. Conclusion：Geniposide has dual regulatory effect on the pinocytosis and phagocytosis of RAW264.7 cells.

Abstract:Objective：To investigate the activation of monocytes（THP-1）by exogenous interleukin 18（IL-18）and the proliferation，metastasis and invasion of lung cancer A549 cells by THP-1 induced by IL-18 activation influences. Methods：The role of tumor associated macrophages（TAMs）in tumor microenvironment in vivo was simulated by IL-18 induced THP-1 in vitro. The chemotactic effect of IL-18 on mononuclear cell THP-1 was detected by Transwell method. Exogenous IL-18 stimulated THP-1 cells and detected the proportion of M1 and M2 TAMs；IL-18-activated TAMs and lung cancer cells A549 were co-cultured. The changes of proliferation，migration and invasion of co-cultured A549 cells were detected by cloning experiment，cell scratch test and cell invasion assay. The morphology of A549 cells after co-culture was observed by scanning electron microscope. The expressions of epithelial mesenchymal transition（EMT）marker proteins E-cadherin，N-cadherin，Snail and Slug in A549 cells were detected by qRT-PCR and Western blot. Results：IL-18 had a significant chemotactic effect on THP-1 cells. Exogenous IL-18 stimulated the THP-1 cell phenotype from M2 to TAMs. IL-18 induced THP-1 promoted the proliferation，migration and invasion of A549. Scanning electron microscopy showed that THP-1 promoted the growth of pseudopod of A549 cells. At the same time，the expression of E-cadherin was inhibited，and the expression of N-cadherin，Snail and Slug was promoted，indicating that IL-18 induced THP-1 activated EMT in A549 cells. Conclusion：In lung cancer，IL-18 can activate THP-1 and promote their activation. Activated THP-1 promotes the proliferation，migration and invasion of lung cancer A549 cells by activating EMT.

Abstract:Objective：To study the change of plasma lipoprotein-associated phospholipase A2（Lp-PLA2）concentration in patients with acute myocardial infarction（AMI）and its correlation with cardiac function after AMI. Methods：Ninety consecutive AMI patients who met the study criteria were enrolled. The plasma Lp-PLA2 levels were measured immediately after admission（T0），the next day（T1），the third day（T2），and the seventh day（T3），N-terminal pro brain natriuretic peptide（NT-proBNP）was measured one month after AMI（T4），left ventricular ejection fraction（LVEF）was measured by two-dimensional echocardiography at T1 and T4. Pearson correlation analysis was used to analyze the correlation between Lp-PLA2 and LVEF or NT-proBNP. In addition，48 healthy volunteers and 43 stable angina pectoris patients were enrolled as the controls. Results：The level of plasma Lp-PLA2 showed dynamic changes after AMI，it was increased significantly after the onset of the disease，but decreased to normal level after 7th days of admission［（61.42±36.99）mg/L vs.（49.83±27.17）mg/L，P > 0.05］. The correlation analysis showed that there was no significant correlation between on-admission plasma Lp-PLA2 and C-reactive protein level（r=0.06，P > 0.05）or between Lp-PLA2 and cardiac troponin T level（r=-0.07，P > 0.05）. Further studies showed that there was no significant correlation between plasma Lp-PLA2 level at T0 and plasma NT-proBNP level at T4（r=0.16，P > 0.05）or LVEF at T4（r=-0.09，P > 0.05）or the difference of LVEF between T1 and T4（r=0.04，P > 0.05）. Conclusion：The plasma level of Lp-PLA2 showed dynamic changes after AMI，which reflected the unstable state of artery plaques，but it was not related to the recovery of cardiac function after AMI.

Abstract:Objective：This study aims to investigate the expression of long intergenic noncoding RNA 460（linc00460）in gastric cancer and its relationship with clinicopathological features and prognosis of gastric cancer patients. Methods：Differential expression of linc00460 was systematically evaluated based on the Cancer Genome Atlas（TCGA）gastric cancer database. Then 88 patients who underwent gastric cancer surgery were included to verify TCGA results by RT-qPCR. The linc00460 expression in AGS，HGC-27，MKN45，MGC-803，and normal gastric epithelial GES-1 were detected by RT-qPCR. Gene set enrichment analysis（GSEA）was used to predict the regulation genes of linc00460. AGS cells were transfected with pcDNA-linc00460 and pcDNA3.1，respectively. Western blot was used to detect the expression of related proteins and CCK-8 method to detect the proliferation of cells in the two groups. Results：We found and confirmed that linc00460 was highly expressed in gastric cancer tissue samples and cells（P < 0.05）；moreover，high linc00460 expression was associated with M-stage；there was a strong negative correlation between the expression of linc00460 and the overall survival of patients with gastric cancer（P < 0.05）；furthermore，univariate and multivariate analysis of the Cox proportional hazards model confirmed that linc00460 was an independent predictor of gastric cancer-related survival（P < 0.05）. The linc00460 high-expression samples in TCGA GC datasets were enriched in the regulation of DNA damage response signal transduction by P53 class mediator，the positive regulation of signal transduction by P53 class mediator，the regulation of meiotic nuclear division and the positive regulation of nuclear division gene sets. The expression level of linc00460 in the pcDNA-linc00460 transfected AGS cells was significantly higher than that of pcDNA3.1 group（P < 0.05）. Western blot results showed that linc00460 could down-regulate the P53 and up-regulate Bcl-2 and Cyclin B1 expressions. CCK-8 results showed cell proliferation in AGS cells of the pcDNA-linc00460 group was significantly higher than that in the pcDNA3.1 group（P < 0.05）. Conclusion：Linc00460 is highly expressed in gastric cancer tissues，and patients with high linc00460 level result in a poor prognosis. Linc00460 may promote the growth of gastric cancer cells by up-regulating the P53 signaling pathway，and linc00460 may be involved in the malignant progression of gastric cancer.

Abstract:Objective：To explore the sequence variation of the promoter region（-436~+267）on stimulator of interferon genes（STING）of children. Methods：The sequences of the promoter region of 48 children were amplified by PCR and then sequenced. The luciferase reporter plasmid pGL-3/STING（TT）and pGL-3/STING（CC）were constructed to transfect human embryonic kidney（HEK）-293T cells， and then the luciferase gene expression was detected and the relative luciferase activity unit（RLU）was calculated. Results：A new single nucleotide polymorphism（SNP）T/C（-401）in STING promoter region was identified. The frequencies of genotype TT，TC，and CC were 45.8%，37.5%，and 16.7% respectively. Restriction endonuclease analysis and DNA sequencing verified the successful construction of the plasmid pGL-3/STING（TT）and pGL-3/STING（CC）including the SNP. Compared with plasmid pGL-3/STING（TT），pGL-3/STING（CC）exhibited a weaker promoter activity. Conclusion：T/C（-401）in STING promoter region is a new SNP. The genotype of this new SNP may reduce the activity of STING promoter，and consequently affect the transcription and expression of STING gene，and affect the body’s susceptibility to disease.

Abstract:Objective：To observe the expressions of human trophoblast cell surface antigen 2（TROP2）and vascular endothelial growth factor receptor 2（VEGFR2）in triple negative breast cancer（TNBC），and explore their correlation with clinicopathological factors and prognosis. Methods：The expressions of TROP2 and VEGFR2 in 151 cases of TNBC，137 cases of non-TNBC，48 cases of benign breast tumor and 48 cases of normal breast tissues were detected by immunohistochemical method（two-step method）. Results：TROP2 and VEGFR2 were expressed in the cell membrane and cytoplasm of breast cancer cells，respectively. The TROP2 and VEGFR2 protein was highly expressed in TNBC，and their positive rates were higher than those in non-TNBC，benign breast tumor and normal breast tissues. The co-expression of TROP2 and VEGFR2 in TNBC was higher than that in non- TNBC，and the difference was statistically significant. The positive expressions of TROP2 and VEGFR2 in TNBC tissues and the high co-expression of TROP2 and VEGFR2 in TNBC were associated with the histological grade，clinical stage and metastasis of patients. The high co-expression of TROP2 and VEGFR2 proteins is an independent prognostic index of TNBC. Conclusion：TROP2 and VEGFR2 can be used as a new molecular target for TNBC，which can be expected to facilitate the prognosis evaluation and immune-targeting treatment of TNBC.

Abstract:Objectives：To explore the diagnostic value of p16/Ki-67 dual stain cytology for high-grade squamous intraepithelial lesion（HSIL）in patients with positive atypical squamous cell of undetermined significance/high risk human papillomavirus（ASCUS/HPV）. Methods：A total of 283 cases diagnosed as HSIL with positive ASCUS/HPV were selected between September，2017 and December，2018 from the Department of Gynecology，the First Affiliated Hospital of Nanjing Medical University. Colposcopy and biopsy were performed on all patients to analyze the risk factors of HSIL. And the diagnostic value of p16/Ki-67 dual stain cytology was compared with that of HPV genotype. Results：In total，88 of the 283 ASCUS/HPV positive cases were diagnosed with HSIL. Univariate logistic analysis indicated that different results of p16/Ki-67 dual stain cytology and HPV genotype test showed different HSIL rates，which were statistically significant（P＜0.01）. Ages of the patients did not contribute to the variation of HSIL incidences（P＞0.05）. Multivariable logistic analysis showed that both HPV genotype and p16/Ki-67 positives were risk factors for HSIL（P＜0.001）. The sensitivity，negative predictive value and area under ROC of p16/Ki-67 dual stain cytology were higher than those of HPV genotype test（P＜0.05）. Conclusion：p16/Ki-67 dual stain cytology positive is a warning sign of HSIL for patients with positive ASCUS/HPV，the screening efficacy of which is better than that of HPV genotype test. It provides a new option for cervical lesion screening.

Abstract:Objective：To investigate the clinical value and significance of high risk human papilloma virus（HPV）E6/E7 mRNA test in patients with HPV infection. Methods：Between January 2017 and June 2017，in Center of Diagnosis and Treatment for Cervical Disease，200 patients with HPV infection in cervical screening received high-risk HPV E6/E7 mRNA test and pathological biopsy under colposcopy. We analyzed the E6/E7 mRNA detection rate of positive cases，and the E6/E7mRNA expression in different pathological diagnosis. Results：The E6/E7 mRNA detection rate was 60% in the 200 patients. The highest E6/E7 mRNA positive expression in the high-grade squamous intraepithelial lesion group（HSIL）had significant differences with the low-grade squamous intraepithelial lesion group（LSIL）and the nomal group. The mRNA sensitivity was 97.9% and negative predictive value was 98.8%（P < 0.05）. The sensitivity，specificity，positive predictive value and negative predictive value were lower in the LSIL group. Conclusion：We could shunt the patients with HPV infection，avoid excessive treatment and reduce the patient’s psychological burden byHR-HPV E6/E7 mRNA test. High-risk HPV E6/E7 mRNA test has high clinical value in diagnosis of HSIL during cervical screening.

Abstract:Objective：To investigate the serum level of stanniocalcin-1（STC1） in patients with stable chronic obstructive pulmonary diseases（COPD） and to explore the relationship between STC1 and severity of the disease. Method：A total of 55 patients with definite stable COPD and 24 healthy controls were enrolled in this study. Serum STC1 was measured by enzyme-linked immunosorbent assay（ELISA），and the correlation between STC1 and COPD was analyzed. Results：①Serum STC1 levels were significantly lower in patients with COPD than healthy controls（P < 0.05）. ②The receiver-operating characteristic（ROC） curve of serum SCT1 was used for threshold identification（AUC=0.659，P=0.025），the optimal cut-off point of serum STC1 was 1 350 pg/mL（sensitivity 81.82%，specificity 50.00%，respectively）. ③Poor association of serum STC1 levels with stable COPD was found，including the GOLD classification，COPD assessment test（CAT） scores and lung functions. Conclusion：Serum STC1 levels were significantly decreased in COPD patients while it is poorly correlated with disease severity. STC1 might have clinical application in stable COPD.

Abstract:Objective：To determine the pathological significance of serum miR-21 expression level in premature respiratory distress syndrome（RDS）infants. Methods：Randomized blood samples were collected from RDS and non-RDS infants in the Neonatal Intensive Care Unit（NICU）of Jiangsu Province Hospital from June to December 2015. Total RNA was extracted from each blood sample and reverse transcribed into complementary DNA（cDNA）. We used real-time reverse transcription polymerase chain reaction（RT-PCR）technique to determine the serum miR-21 expression levels in the RDS and the non-RDS groups（n=30 and n=21，respectively）. Unpaired student t-test was used to compare the significance between groups. Results：The serum miR-21 expression level in the RDS group was significantly higher than that in the control group（P < 0.05）. The miR-21 expression levels in the moderate to severe RDS groups appeared to be higher than that in the mild groups，however，statistic significance was not reached in our study. In addition，the RDS group that received pulmonary surfactant therapy had significant increase in miR-21 expression levels（P < 0.05）. Conclusion：Serum miR-21 expression level is closely related to the development of infant RDS，suggesting that miR-21 is involved in the pathogenesis of infant RDS.

Abstract:Objective：To evaluate the antiplatelet efficacy of ticagrelor in patients after off-pump coronary artery bypass graft（OPCABG）surgery and observe its clinical outcomes. Methods：A total of 90 patients undergoing OPCABG successfully in the First Affiliated Hospital of NMU during May 2015 to June 2016 were enrolled in this study. Patients were randomly divided into three groups. In the first day after surgery，group A was given clopidogrel（75 mg，1/day，oral）；group B was given ticagrelor（90 mg，2/day，oral）；group C was given half amount ticagrelor（45 mg，2/day，oral）. The three groups took aspirin standard dose（100 mg，1/day）at the same time. In 5 to 7 days after OPCABG，platelet inhibition（PI）rate induced by arachidonic acid（AA）and adenosine diphosphate（ADP）（PIADP and PIAA）by TEG was monitored to compare antiplatelet efficacy of the three groups. Major adverse cardiac events（MACE），bleeding events and related adverse reactions were also compared between the groups to evaluate the clinical effect. Results：Compared with group A，PIADP（%）of group B and group C were significantly increased（83.8 ± 17.6 vs. 60.5 ± 25.6，P=0.001；77.9 ± 24.6 vs. 60.5 ± 25.6，P=0.013，respectively）. There was no obvious difference of PIADP（%）between group B and group C（77.9 ± 24.6 vs. 83.8 ± 17.6，P > 0.05）. The three groups were not seen obvious MACE. Compared with group A，incidence of bleeding events in group B was increased，but there was no statistically significant difference（23.3% vs. 16.6%，P > 0.05）. Conclusion：Ticagrelor treatment as antiplatelet therapy in patients after OPCABG was effective and safe，and its antiplatelet efficacy was superior to clopidogrel.

Abstract:Objective：A localization system for multiple magnetic targets based on magnetic sensor rotation detection was studied，which was aimed at realizing human multiple capsule endoscopy positioning and nano-magnetic drug testing. Methods：A three-axis tunneling magnetoresistive sensor was used to make a circular motion near the magnetic source. Three sets of magnetic signals were acquired and transferred by three-dimensional coordinates. The position of the permanent magnet was solved using a positioning algorithm. Results：Through the preparation of LabVIEW PC program，the trajectory of two magnetic target tracking and motion curve storage and playback were achieved. Conclusion：The detection system has the advantages of good real-time，high precision and low cost，it has the potential application value for the positioning of multiple capsules in the human body and the detection of nano-magnetic drugs.

Abstract:Objective：To compare the predictive performance of constructing the early prognostic model of acute paraquat（PQ）poisoning between support vector machine（SVM）and logistic regression. Methods：A total of 152 patients with acute PQ poisoning were collected and the clinical results were observed for 2 months. The patients were divided into two groups with a 3∶2 ratio by the random numerical table method. One group with a total of 91 cases was used as a training sample for selecting variables and establishing predictive models. Another group with a total of 61 cases was used as a validation sample to evaluate the predictive effect of the model. SVM and conventional logistic regression was used as the modeling method. Results：The prediction accuracy of the kernel，polynomial，sigmoid kernel and radial basis function nuclear SVM model was 77.92%，74.03%，75.32% and 79.22% respectively，when being tested by the validation group. The results of performance comparison showed that SVM models performed better than logistic regression model；RBF-SVM was the best among all the models with a sensitivity of 87.5% and a specificity of 70.6%. Conclusion：SVM model could preferably integrate all kinds of prognostic information of PQ poisoning patients，and the established model had better prediction ability，providing a new method for predicting the prognosis of patients with PQ poisoning.

Abstract:The polycystic ovary syndrome（PCOS）is defined as a complex endocrine syndrome，which exhibits chronic ovulatory disorders，hyperandrogenism，insulin resistance and some other metabolic disorders. However，the mechanisms underlying these various clinical signs and symptoms are still not entirely known，which causes difficulties for clinical diagnosis and treatment of PCOS. Along with the development of metabonomics technologies，the available technique to detect and comprehensively analyze the innumerable small-molecule metabolities，may allow to find out the interactions between the symptoms and pathogenesis of PCOS， indicate the origin of these metabolic alterations，and discover new metabolic biomarkers for PCOS. This review summarizes the metabolites associated with PCOS according to recent metabolomic studies，and hopefully，some new biomarkers may serve to predict the progression of the disease and promote clinical individualization of diagnosis and treatment.

Abstract:Pierre Robin sequence（PRS） is a congenital malformation consisting of glossoptosis，retrognathia，airway compromise，often with clefting of the secondary palate. There is no uniform standard for diagnosis and treatment at home and abroad. The measurement of PRS has been an important way to assist diagnosis and treatment over the years. The measurement methods include somatometry，imaging measurement，etc，and the measurement indicators are various. This paper reviews the measurement of PRS.