Abstract:Objective：This study compared the expression spectrum of long noncoding RNA（lncRNA） and messenger RNA（mRNA）in pancreatic tissue between intrauterine growth retardation（IUGR）and normal neonatal mice，and then explored the potential mechanism of islet developmental disorders in IUGR mice and development of diabetes later in their adulthood. Methods：IUGR and normal neonatal mice models were established，whole transcriptome sequencing in pancreas of two groups were performed，and then gene ontology（GO）and Kyoto Encyclopedia of gene and genomes（KEEG）enrichment analysis were conducted. To further analyse the whole transcriptome sequencing information，we selected several lncRNAs with high differential expression for validation，which were also reported to be involved in islet development. Results：Total 294 lncRNAs were differentially expressed between IUGR and normal neonatal mice，among which 83 were up-regulated and 211 were down-regulated. There were 2 000 differentially expressed mRNAs，of which 1 711 were up-regulated and 289 were down-regulated. The GO analysis of lncRNA target genes and differential mRNAs showed：biological pathways（BP）are concentrated in cellular processes，biological regulation and metabolic processes；cell components（CC）are concentrated in cell and organ processes；molecular function（MF）mainly focuses on integration，catalytic activity，transportation activity，etc. KEEG enrichment analysis highlights the involvement of PI3K-Akt pathway，MAPK pathway and Foxo pathway. Through verified experiments，the selected lncRNA Snhg12 and Rian showed significant differences between IUGR mice and normal mice，which is consistent with the sequencing results，and the expression were regulated by glucose concentration. Conclusion：This study analyzed and partially verified the lncRNA and mRNA differentially expressed in IUGR，which is helpful to explore the potential mechanism of islet developmental disorders and later adult diabetes in mice born with IUGR，especially the role of lncRNAs.

Abstract:Objective：This study aimed to investigate the promotion effect of low molecular weight hyaluronic acid（LMW-HA）on phenotypic transformation of mice cardiac fibroblasts and the role of CD44 and S100A4 in this process. Methods：Cardiac fibroblasts（CFs）were isolated and cultured from neonatal ICR miceandthen stimulated by LMW-HA. The proliferation of CFs was measured by CCK-8 and EdU. Western blot（WB），quantitative RT-PCR and immunofluorescence were used to determine myocardial fibrosis. Nuclear and cytoplasmic proteins of CD44 and S100A4 were extracted to specify the nuclear translocation by WB and immunofluorescence. Above indicators were measured again after CFs treated with the CD44 inhibitor BRIC-235. Results：CCK-8 and EdU determined that the optimal concentration of LMW-HA stimulation was 0.8mg/mL. WB，qRT-PCR and immunofluorescence showed that LMW-HA stimulation significantly increased the level of myocardial fibrosis markers（α-SMA and Collagen 3），but CD44 remained unchanged，and S100A4 gradually increased with time.The proteins of CD44 and S100A4 were transferred into the nucleus at the same time. BRIC-235 can inhibit these changes. Conclusion：LMW-HA can promote the differentiation of micecardiac fibroblasts，which is regulated by the nuclear translocation of CD44 and S100A4 and the activation of downstream signaling pathway.

Abstract:Objective：This study aims to explore the dynamic changes in the process of radiation-induced pulmonary fibrosis（RIPF） of B10 cells. Methods：30 C57BL/6 mice，6-8 weeks old，were randomly divided into non-irradiated（control）group，2 days after irradiation（IR2d），14 days after irradiation（IR14d），3 month after irradiation（IR3m）and 5 month after irradiation（IR5m）group.The radiation-induced pulmonary fibrosis model was established by using ionizing radiation. HE staining and Masson staining were used to observe the pathological changes of lung tissue，and the expression of α-SMA was observed by immunohistochemistry. Flow cytometry was used to detect the infiltration changes of B10 cells in lung tissues and spleens in the RIPF model. Results：The lung tissues of mice showed inflammatory response in the early stage after chest irradiation. Furthermore，the alveolar structure destruction and interstitial filling began to appear 3 months after irradiation，and a large amount of collagen was filled 5 months after irradiation. The expression of α-SMA was significantly higher than that of the control group，and pulmonary fibrosis was more severe. The proportion of B10 cells in lung tissue was significantly increased at 2 d and 14 d after chest irradiation compared with the non-irradiated group，while was significantly lower in IR3m group than in IR2d group，B10 cells in the spleens began to increase at IR2d，and reached a peak at IR14d，and was significantly lower in IR3m group than in IR2d group and IR14d group. Conclusion：There were B10 cells infiltration in the process of radioactive pulmonary fibrosis，with a trend of early increase and late decrease.

Abstract:Objective：This study aims to investigate whether conditioned medium from human amnion-derived mesenchymal stem cells（hAMSC-CM） promotes angiogenesis of human umbilical vein endothelial cells（HUVEC） via circRNA-0001649（circ-0001649）. Methods：HUVECs were divided into control group，conditioned medium stimulation group（CM-group），conditioned medium stimulation after knockdown of circ-0001649 group（si-circ-0001649+ CM group） and knockdown control with conditioned medium stimulation group（si-NC+CM group）. And the angiogenic and migratory abilities of HUVECs in each group were then detected，as well as the expression of VEGFA and MMP9. After that，inhibition of miR-203a followed by down-regulation of circ-0001649 in HUVECs treated with hAMSC-CM were functional performed to confirm the relative mechanism. Results：The hAMSC-CM significantly increased the angiogenic and migratory capacity of HUVECs，the protein levels of VEGFA and MMP9，and the expression of intracellular circ-0001649. Knockdown of circ-0001649 inhibited the effect of hAMSCs-CM on the angiogenesis and migratory of HUVECs. Bioinformatic analysis and a series of experiments revealed that circ-0001649 could promote the angiogenic and migratory ability of hAMSC-CM on HUVECs by binding miR-203a and then reduced the inhibition of miR-203a on VEGFA and MMP9. Conclusion：The hAMSC-CM up-regulated circ-0001649 in HUVECs，which could binding miR-203a in a ceRNA manner，and promoted angiohenesis by increasing VEGFA and MMP9 expression.

Abstract:Objective：This study aims to prepare a human anti-c-Met full-molecule antibody（c-Met-IgG1）conjugated with recombinant analgesic-antitumor peptide（rAGAP） to analyze their biological characteristics and detect their effects on the biological behavior of ovarian cancer cells. Methods：Using the human anti-c-Met antibody gene stored in our laboratory as a template，construct the c-Met-IgG1-rAGAP eukaryotic expression vector was constructed and transfected it into eukaryotic cells to obtain the antibody conjugate. The immunological properties of c-Met-IgG1 and c-Met-IgG1-rAGAP were evaluated by methods such as affinity detection，immunofluorescence，and flow cytometry. CCK-8，scratch test，and Transwell invasion test were used to detect the effects of antibodies on the proliferation，migration and invasion of ovarian cancer cells. Results：Two antibodies，c-Met-IgG1 and c-Met-IgG1-rAGAP，which can specifically bind to c-Met protein，were successfully prepared；c-Met-IgG1 and c-Met-IgG1-rAGAP proved in a series of experiments can inhibit the proliferation，migration and invasion of the c-Met-positive ovarian cancer cell HO8910，and the inhibitory effect of c-Met-IgG1-rAGAP on ovarian cancer cells is higher than that of c-Met-IgG1，while these effects were not observed in the c-Met-negative IOSE386 cells. Conclusion：The research results show that c-Met-IgG1-rAGAP can target c-Met-positive ovarian cancer cells and has anti-tumor activity，which can lay a research foundation for exploring molecular targeted treatment of ovarian cancer.

Abstract:Objective：This study aims to observe the effects and mechanisms of Wnt5a on the formation of neutrophil extracellular trap（NET）. Methods：Neutrophils were stimulated with 50 ng/mL Wnt5a for 3 h，1 μg/mL Porphyromonas gingivalis （P.gingivalis）lipopolysaccharide（LPS） and 1 μg/mL Escherichia coli（E.coli） LPS were served as positive controls. Formation of NET was observed by immunofluorescence staining，and extracellular DNA was quantified by Sytox Green fluorescence staining. Levels of reactive oxygen species（ROS） were detected by flow cytometry，and activations of phosphatidylinositol 3-kinase（PI3K）/Akt，mitogen-activated protein kinase 1/2（MEK1/2）and extracellular regulated protein kinases 1/2（ERK1/2） were explored by Western blot. Results：Compared with neutrophils without stimulations，Wnt5a stimulation led to the formation of NET and the increased levels of extracellular DNA（P < 0.05）. In addition，levels of ROS and phosphorylations of PI3K/Akt，MEK1/2 and ERK1/2 were significantly increased in the cells treated with Wnt5a（P < 0.05）. Conclusion：Wnt5a might contribute to the formation of NET depending on the production of ROS and the phosphorylations of PI3K/Akt，MEK1/2 and ERK1/2.

Abstract:Objective：The dental pulpitis model of α7 nAChR gene knockout mice was established to provide an experimental model for studying the mechanism of α7 nAChR in the occurrence and development of dental pulpitis. Methods：Sixteen α7 nAChR knockout mice（knock out，KO）and 16 wild type（wild type，WT）in C57BL/6 were selected，and a hole was drilled on the maxillary first molars to expose dental pulp to constructed dental pulpitis model. The mice were sacrificed on 1 day，3 days，7 days after operation respectively. HE staining and immunohistochemistry were carried out for detecting NOD-like receptor protein 3（NLRP3）. Results：HE staining showed blood congestion and tissue edema around the medullary foramen in α7 nAChR KO mice and WT mice at 1 day after medullary cavity exposure. On 3 days after the exposure of the dental pulp cavity，the inflammatory cells were aggregated，reached to the bottom of the crown pulp in α7 nAChR KO mice，while were mainly around the perforationin the pulp tissue in WT mice. On 7 days，pulpitis cells of α7 nAChR KO mice were infiltrated into root pulp，while inflammatory pulp cells of WT mice were still concentrated mainly in the crown pulp. The expression of NLRP3 in pulpitis of α7nAChR KO mice was significantly higher than that of WT mice by immunohistochemical staining（P < 0.05）. Conclusion：The pulpitis model of α7 nAChR gene knockout mice was successfully constructed，and the pulpitis infiltration of α7 nAChR gene knockout mice was significantly faster than that of WT mice.

Abstract:Objective：Two gold nanoparticle-microbubble composites with different structures were constructed，which can be used as ultrasound imaging contrast agents and for photothermal conversion using the surface plasmonic excitonic effect of gold nanoparticles. Methods：One is gold encapsulated microbubble（AuMB）with gold nanoparticles encapsulated in a microbubble gas core，and the other is polyethyleneimine-gold loaded microbubble（PEI-AuMB）with gold nanoparticles loaded onto the microbubble surface by covalent bonding. These gold nanoparticle-microbubble composites were characterized for their physicochemical properties，and their ultrasound contrast capabilities were evaluated by in vitro ultrasound imaging. Their photothermal conversion ability was evaluated by in vitro photothermal experiments. Results：Both composite microbubbles showed better ultrasonography ability than blank microbubble（MB），among which PEI-AuMB showed better performance in the duration of ultrasonography enhancement；both composite microbubbles also showed superior photothermal conversion ability than gold nanoparticles alone，and AuMB had better photothermal therapeutic potential. Conclusion：All these results demonstrates that gold nanoparticle-microbubble composites are promising carrier systems for therapeutic integration.

Abstract:Objective：In order to evaluate the microbial contamination of different departments and explore the diversity of living bacterial community，the method of propidium monoazid（PMA） pretreatment combined with high-throughput sequencing technology was used to detect the living microbial community in the dental unit waterline（DUWL）. Methods：A total of 33 dental chair units（DCU）were selected from the Affiliated Stomatological Hospital of Nanjing Medical University，with 23 from endodontics department（group E）and 10 from periodontics department（group P）. Before patients arrived，water samples were collected from each DCU using a three-in-one air/water syringe. After PMA pretreatment，the total DNA of living bacteria was extracted. After PCR amplification with bacterial universal primers，the macrogenomic library was constructed for high-throughput sequencing and bioinformatics analysis. Results：Samples were pretreated with 1 μL of PMA（20 μmol/L）for 10 min and subsequently exposed to light irradiation for 5 min，which were optimal for detecting viable bacterial in samples of DUWL. The output water of DUWL was associatied with a high bacterial diversity and identified potential bacterial pathogenic sequence. The statistically different phyla between the two groups were Cyanobacteria，Actinobacteria，Chlamydiae，Aerophobetes，Chloroflexi，Bacteria_d_norank_k__unclassified，TM6（P < 0.05）. The statistically different genera between the two groups were Legionella，Methyloversatilis，Obscuribacterales__norank（P < 0.05）. Conclusion：The occurrence of potentially pathogenic microorganisms in these DUWLs demonstrated a potential infectious risk for both dental health care personnel and patients. Attention should be paid to the diversity of DUWL bacterial communities . It is necessary to proceed to regular water quality control of DUWLs.

Abstract:Objective：This study aims to investigate the value of bile acid profile in diagnosis of intrahepatic cholestasis of pregnancy（ICP）and asymptomatic hypercholanemia of pregnancy（AHP）. Methods：Total 100 cases of normal pregnancy，60 cases of ICP and 30 cases of AHP were selected. The content of bile acid in each group was analyzed by high performance liquid chromatography tandem mass spectrometry（HPLC-MS/MS）. In addition，the orthogonal projections to latent structures（OPLS-DA）model was developed to explore the differences of bile acids between the ICP and AHP groups. The diagnostic effiency of the differential bile acids and the combined indicators was analyzed by receiver operating characteristic（ROC） curves. Results：Except for chenodeoxycholic acid（CDCA），the contents of 15 kinds of bile acids among the three groups were statistically significant（P < 0.05）. According to the OPLS-DA model，the differential bile acids in ICP group and AHP group were taurocholic acid（TCA），taurochenodeoxycholic acid（TCDCA），glycocholic acid（GCA），glycohenodeoxycholic acid（GCDCA），tauroursodeoxycholic acid（TUDCA）. ROC results showed that TCA had the highest diagnostic efficiency with area under curve（AUC） of 0.808. The combination of TCA，TCDCA，GCDCA and TUDCA was more effective in the diagnosis of ICP. The AUC was 0.967，the sensitivity was 90.6%，and the specificity was 98.0%. Conclusion：There are differences in bile acid profile between ICP and AHP. The differential bile acids found by OPLS-DA model can be used as an important method for differential diagnosis between ICP and AHP.

Abstract:Objective：This study aims to investigate the risk factors of preterm twin pregnancy and establishment of predictive model. Methods：Twin pregnant women delivered in the Affiliated Obstetrics and Gynaecology Hospital of Nanjing Medical University from January 2017 to December 2019 were analyzed retrospectively，who were divided into preterm group（<37 weeks）and term group（≥37 weeks）according to the gestational week of delivery. Univariate analysis was used to analyze the general conditions，pregnancy complications of the two groups. Multivariate logistic regression analysis was used to analyze the independent risk factors of preterm in twin pregnancy. ROC curve was used to evaluate the predictive value. Results：A total of 1 783 pregnant women with twin pregnancy，856 in the preterm group，and 927 in the term group were collected，the incidence of preterm birth was 48%. Causes of preterm labor in the twin pregnancy were listed at below：iatrogenic preterm labor accounted for 43%，premature rupture of membranes accounted for 29%，and spontaneous preterm labor accounted for 28%. Irregular prenatal examination，monochorionic twins，discordant twins，cicatrical ulerus，premature rupture of membranes，hypertension in pregnancy，abnormal amniotic fluid，intrahepatic cholestasis，intrauterine distress，and chorioamnionitis in the preterm group were higher than those in the control group，and the differences were statistically significant（P < 0.05）. Multivariate logistic regression analysis showed that discordant twins，irregular prenatal examination，monochorionic twins，intrahepatic cholestasis，abnormal amniotic fluid，hypertensive disorder complicating pregnancy， premature rupture of membranes，intrauterine distress，chorioamnionitis and cicatrical ulerus were the independent risk factors of preterm twin pregnancy. The ROC curve verified the high predictive efficiency of the model. Conclusion：The occurrence of preterm birth in twin pregnancy is related to a variety of maternal and fetal risk factors. The incidence of preterm twin pregnancy is fairly high and could be predicted from the risk factors.

Abstract:Objective：This study aims to investigate the expression level of HER2/Ki67 is association with efficacy of anthracycline chemotherapeutic drugs and clinical prognosis in invasive breast cancer. Methods：The expression level of HER2 and Ki67 in invasive breast cancer were predicted by database. The mRNA expression of HER2 in 53 cases of invasive breast cancer FFPE was detected by Nanostring nCounter technique，and the efficacy of anthracycline chemotherapeutic drugs was analyzed. Total 222 cases of invasive breast cancer tissue samples from 2008 to 2012 were collected to analyze the protein expression level of HER2 and Ki67，and the correlation analysis was conducted based on the clinicopathological data and the chemotherapy efficacy of the patients. Results：GEPIA database analysis showed that there was a correlation between the HER2 mRNA and Ki67 mRNA. The expression level of HER2 mRNA in 53 cases of invasive breast cancer was detected by Nanostring nCounter. The positive rate was 16/53（30.19%）and the negative rate was 37/53（69.81%）. Among the HER2 positive patients，chemotherapy was effective in 3 cases（18.80%）and ineffective in 13 cases（81.20%）. In the HER2 negative patients，chemotherapy was effective in 19 cases（51.35%）and ineffective in 18 cases（48.65%）. The results of the clinicopathological data of 222 cases of invasive breast cancer showed that the invasive breast cancer patients with HER2-/KI67+ predicted better chemotherapy efficacy and prognosis. Conclusion：In patients with HER2-negative invasive breast cancer，the high expression of Ki67 indicates a better effect of chemotherapy，while the patients with HER2-/Ki67+ predict a better survival time.

Abstract:Objective：This study aims to explore METTL3 combined with programmed death cell ligand 1（PD-L1）detection in predicting the therapeutic effect and prognosis of NSCLC patients with programmed death cell protein 1（PD-1）inhibitors treatment. Methods：Primary lesion tissues from NSCLC patients were collected. The protein expression of PD-L1 and METTL3 was detected by immunohistochemistry，and the association of METTL3 with the effect of PD-1 inhibitor treatment and prognosis in patients with NSCLC was analyzed. Results：A total of 228 cases were collected. Among them，the protein expression of METTL3 was related to the pathological type，Eastern American Oncology Collaborative Group（ECOG）score，therapeutic plan，therapeutic effect and Response Evaluation Criteria in Solid Tumors（RECIST）factors. Patients with high METTL3 protein expression had a median progression-free-survival（PFS）of 12.33 months，while those with low or no METTL3 protein expression had a median PFS of 6.50 months. In NSCLC patients treated with PD-1 antibody，high METTL3 protein expression was associated with better PFS. The median overall-survival（OS）of patients with high METTL3 proteine xpression was 23.54 months，and OS of patients with low or no METTL3 protein expression was 20.93 months. Patients with high METTL3 protein expression were associated with better OS. Conclusion：METTL3 protein may be a new target for predicting the efficacy of PD-1 inhibitors treatment in patients with NSCLC.

Abstract:Objective：This study aims to analyze the risk factors related to apolipoprotein E（APOE）gene polymorphism and advanced glycation end products in elderly patients with coronary heart disease. Methods：The distribution of APOE gene polymorphism in 46 elderly patients with coronary heart disease and 44 non-coronary heart disease patients（control group）was observed，and the plasma levels of carboxy methyl lysine（CML）and soluble receptor for advanced glycation end products（sRAGE）were determined by ELISA. Results：The allele frequency of ApoE ε4 in coronary heart disease group was significantly higher than that of the control group（P < 0.05），and the level of plasma CML and sRAGE in coronary heart disease group was higher than that of the control group，with no statistically significant difference. The plasma CML and sRAGE levels of APOE ε4 carriers in the coronary heart disease group were not significantly different from those in the control group.The risk factors of coronary heart disease in elderly patients with APOE ε4 carriers were 3.511 times that of non-carriers（P < 0.05），but CMLand sRAGE levels were not significantly associated with an increased risk of coronary heart disease. Conclusion：With age，APOE ε4 allele remained an independent risk factor for coronary heart disease in elderly patients，and there was no significant correlation between advanced glycation end products level and coronary heart disease in elderly patients.

Abstract:Nasopharyngeal carcinoma（NPC） is closely related to Epstein-Barr virus（EBV），which mostly occurs in south China and Southeast Asian countries. Because of the anatomical limitation of NPC and its sensitivity of radiotherapy，radiotherapy can achieve satisfactory results for early patients，However，most patients with NPC are locally advanced when they are diagnosed，for these patients，comprehensive treatment based on radiotherapy has gradually become the main means of treatment. How to improve the treatment effect and life quality of patients with NPC is still a big challenge. In this paper，the development of chemoradiotherapy and the choice of chemotherapy regimens，as well as the development of new technologies such as targeted therapy，immunotherapy and surgery are discussed.

Abstract:Congenital heart disease is thought to be closely related to thyroid hormones. Surgery for congenital heart disease with cardiopulmonary bypass can lead to a significant decrease in thyroid hormones，and hypothyroidism can also affect recovery from heart surgery. Postoperative thyroid hormone replacement therapy as a feasible method，but it does not provide clear evidence of the value of thyroid hormone therapy. The routine application of thyroid hormone after cardiac surgery to correct Euthyroid sick syndrome（ESS）in children has not been established. This paper discusses the relationship between congenital heart disease and hypothyroidism to explore the feasibility of thyroid hormone replacement therapy.

Abstract:Objective：Exosomes as vesicles are secreted by a variety of cells in the human body，they exist in serum，urine，saliva and follicular fluid. These molecules play important regulatory roles in many biological and pathological processes，such as cell differentiation，proliferation/growth，apoptosis，angiogenesis and inflammation. The microRNA（miRNA）is a small noncoding RNA，about 17-24 nt，it mediates posttranscriptional regulation by binding to complementary sequences on messenger ribonucleic acid（mRNA）and promoting mRNA degradation or translation inhibition. miRNA is closely related to the occurrence and development of polycystic ovary syndrome，abortion，endometriosis and ovarian cancer. Therefore，exosomal miRNA has a wide range of clinical application potential in the monitor and treatment of disease.