Abstract:The emergence and outbreak of 2019 novel coronavirus（2019-nCoV）have posed a great challenge. Vaccination is an effective way to protect the susceptible from the spread of infectious diseases. The purpose of this essay is to review the inactivated vaccine，subunit vaccine and DNA vaccine research progress of 2002 severe acute respiratory syndrome（SARS）coronavirus and 2012 middle east respiratory syndrome（MERS）coronavirus. Combined with the coronavirus vaccine progress，the history of vaccinia vaccine，and the characteristics of coronaviruses，we provide some potential approachesof vaccine development for 2019-nCoV.

Abstract:The novel coronavirus pneumonia，which spread rapidly across China and other countries，has been proved the obviou sinfectivity and harm to human health. Since there is no specific antibody against the novel coronavirus in the body，it is highly spread after human infection，which threat to human health and public health security in the world. At present，there is no available vaccine and specific antiviral drugs for the novel coronavirus pneumonitis，so the antibody development mediated prevention and clinical treatment is particularly important. This review evaluates the research on antibody detection，neutralization antibody and subunit vaccine to the novel coronavirus pneumonia，which aimed to focus on the antibody strategies against the novel coronavirus，providing new ideas for the prevention and control of the novel coronavirus pneumonia.

Abstract:Objective：This study aims to investigate the relationship between RNA binding protein 7（RBM7） and P21 via overexpressing and knocking down RBM7 in human breast cancer cell BT474. Methods：BT474 cells were respectively transfected with RBM7 overexpressing，knocking down lentivirus（experimental group）and corresponding control lentivirus（control group）. The expression of RBM7 was verified after BT474 stably transfected with RBM7 letivirus by the qRT-PCR and Western blot assays. The effects of RBM7 overexpression or knock down on breast cancer proliferation were evaluated by using CCK-8 assays. At the same time，the flow cytometry assays were used to detect the cell cycle distribution of RBM7 overexpression or knock down. The qRT-PCR and Western blot assays were used to investigate the expression of P21 following the variation of RBM7. Furthermore，the relationship between RBM7 and P21 was studied by RNA binding protein immunoprecipitation（RIP） assays. Results：The green fluorescence was observed by fluorescence microscope when BT474 was transfected with RBM7 overexpression and knock down lentivirus after 48 h. After stably screening，the fluorescence expression of cells was significantly stronger. CCK-8 and the flow cytometry assays showed that overexpressing RBM7 could promote the proliferation of breast cancer cells，while knocking down RBM7 could inhibit the proliferation of breast cancer cells. It was observed that overexpression of RBM7 could downregulate mRNA（P < 0.05）and protein expression of P21，and knockdown of RBM7 upregulated mRNA（P < 0.05）and protein expression of P21 by qRT-PCR and Western blot. It was revealed that RBM7 could directly bind to mRNA of P21 by RNA-binding protein immunoprecipitation（RIP）assays. Conclusion：RBM7 can downregulate the expression of P21 by binding to its transcript in breast cancer cell BT474.

Abstract:Objective：This study aims to investigate the effects of liraglutide（Li） on bleomycin（BLM）-induced pulmonary fibrosis in rat and the potential mechanisms underlying. Methods：SD rats were randomly divided into four groups：control group（Con），BLM group，BLM+Li group and Li group. After BLM（4 U/kg）was intratracheally administered once to induce pulmonary fibrosis，Li（0.2 mg/kg）was subcutaneously injected daily in BLM+Li and Li group while the control group was given the same volume of normal saline. On the day 28，all rats were anesthetized and the lung tissues were rapidly excised. Pulmonary injury and pulmonary fibrosis was evaluated by the hematoxylin-eosin and masson’s trichrome staining. Mast cells were detected with toluidine blue staining. The hydroxyproline content in the lung was detected by alkali hydrolysis assay. Collagen type I alpha 1，collagen type Ⅱ alpha 1，collagen type Ⅲ alpha 1，tumor necrosis factor（TNF）-α，interleukin（IL）-6，IL-1β and transforming growth factor β1（TGF-β1）mRNA were detected by real time quantitative PCR. ELISA was performed to analyze the level of TNF-α，IL-6 and IL-1β in the lung homogenates. E-cadherin and α-smooth muscle actin（α-SMA） expression were evaluated by immunofluorescence. The protein levels of E-cadherin，zona occluden 1（ZO-1），Occludin，α-SMA and TGF-β1 were detected by Western blot. Results：Compared with the BLM group，Li reduced inflammatory cell infiltration，interstitial thickness，collagen deposition，significantly reduced the expression of TNF-α，IL-6，IL-1β and TGF-β1；up-regulated the expression of E-cadherin，ZO-1，Occludin and down-regulated the expression of α-SMA. Conclusion：Li attenuates BLM-induced pulmonary fibrosis in rat，which might be attributed to its effects on alleviating inflammation，suppressing the expression of TGF-β1 and inhibiting epithelial-mesenchymal transition. Li might be a potential therapeutic option for pulmonary fibrosis in the future.

Abstract:Objective：The aim of the study was to investigate the effect of 17β-estradiol（E2）on the expression of RhoA/ROCK pathway in rat colon smooth muscle cells（SMC）. Methods：Sprague-Dawley rat colonic SMCs were isolated and cultured in vitro. Double immunofluorescence staining was used to observe the expression of ROCK1/ERα and α-actin. After stimulation with E2 at different concentrations and different times，the expression of RhoA and ROCK1 were detected by reverse transcription real-time quantitative PCR（qRT）-PCR and Western blot. After stimulation with different concentrations of ROCK inhibitor Y27632 and 17β-estradiol，the expression of ROCK downstream protein was detected by Western blot. The effects of estrogen receptor（ER）inhibitor ICI182780，bovine serum albumin-bound（BSA-E2），ERα selective agonist propyl pyrazole triol（PPT）and ERβ selective agonist diarylpropiolnitrile（DPN）on RhoA/ROCK pathway expression were observed. The ERα and ERβ siRNAs were constructed，and the effects of E2 stimulation on the expression of RhoA and ROCK1 in SMCs were observed. Results：Double immunofluorescence staining showed that ROCK1 and α-SMA co-expressed in rat colon SMC. Treatment with 50 nmol/L E2 for 24 h could significantly inhibit the expression of RhoA and ROCK1（P < 0.05）. ROCK inhibitor Y27632 inhibited the expression of downstream proteins p-MYPT1，p-CPI17 and p-MLC in a concentration-dependent manner. The expressions of RhoA and ROCK1 in E2 group and PPT group were significantly lower than those in other groups（P < 0.05）. The expressions of RhoA /ROCK1 in ERα siRNA group were higher than those in control group（P < 0.05）. Conclusion：ROCK1 is localized in rat colonic SMCs，and E2 inhibits the RhoA/ROCK pathway in an ERα-dependant manner．

Abstract:Objective：This study aims to explore the role and mechanism of endothelial progenitor cells（EPC） promoting monocyte/macrophage to type M1 polarization under ischemia and reperfusion. Methods：the peripheral blood of healthy volunteers was obtained，the primary EPC and mononuclear/macrophage were cultured. The model of cell ischemia-reperfusion injury was established in vitro，and the expression of intercellular adhesion molecule -1（ICAM-1），vascular cell adhesion molecule -1（VCAM-1）and E- selectin on EPC surface were detected by flow cytometry. ELISA was used to detect the concentration of ICAM-1，VCAM-1 and E- selectin in the supernatant. Co-culture of EPC and mononuclear/macrophages was performed by Transwell chamber，and the ratio of M1 to M2 type monocytes/macrophages was detected by flow cytometry. Results：There was no significant change in the expression and the secretion of ICAM-1 and VCAM-1 of EPC under the ischemic reperfusion condition，and there was no significant difference between the two groups. The average fluorescence intensity of E- selectin on the surface of EPC was 10.89 in the control group and 33.93 in the ischemia reperfusion group（t=3.895，P=0.018）. The concentration of E- selectin in the EPC supernatant was 3.69 ng/mL in the control group and 18.17 ng/mL in the ischemia-reperfusion group（t=4.568，P=0.010）. Under ischemia reperfusion condition，EPC promoted monocyte/macrophage to M1 type polarization. The proportion of monocyte/macrophage type M1 was 58.83% in control group and was 81.43% in ischemia reperfusion group（t=5.394，P=0.006），E- selectin blockage could inhibit this effect（t=5.950，P=0.004）. Under ischemia reperfusion condition，EPC inhibited monocyte/macrophage to M2 type polarization. The ratio of M2 monocyte/macrophage was 60.57% in control group and was 35.30% in ischemia reperfusion group（t=6.424，P=0.003），E- selectin blockage could inhibit this effect（t=4.179，P=0.014）. Conclusion：Under the condition of ischemia-reperfusion injury，EPC can promote the monocyte/macrophage to M1 polarization through the high expression and secretion of E- selectin. Our research provides a new target for the treatment of ischemia-reperfusion injury.

Abstract:Objective：This stady aims to observe the role of hepatocyte growth factor（HGF） derived from gastric cancer mesenchymal stem cells（GC-MSC）in the proliferation，migration and tumor formation of gastric cancer cells. Methods：The proliferation activity of gastric cancer cells was detected with CCK8 assay. The colony forming ability of gastric cancer cells was analyzed via colony formation assay. Transwell assay and cell wound scratch assay were used to measure the migration ability of gastric cancers. By methods of Western blot and flow cytometry，we detected the expression of programmed cell death-ligand 1（PD-L1）in gastric cancer cells or cancer tissues. BALB/c nu/nu subcutaneous tumor model was constructed to observe tumor formation in vivo. The expression levels of HGF in serum and culture media were determined by ELISA，and protein levels of HGF in tissues of gastric cancer patients and healthy people were detected by immunohistochemistry. Results：The content of HGF in culture medium of gastric cancer mesenchymal stem cells（GC-MSC-CM） was significantly higher than that in culture medium of bone marrow mesenchymal stem cell（BM-MSC-CM）（P < 0.05）. GC-MSC-CM treatment enhanced the proliferation，migration and colony formation of gastric cancer cells，and the promoting effect could be inhibited by HGF blockade. GC-MSC-CM could also promote the expression of PD-L1 in gastric cancer cell lines，and the proportion of PD-L1 positive cells decreased after HGF inhibition. In addition，exogenous HGF could also promote proliferation and PD-L1 expression of gastric cancer cells. The serum HGF levels in gastric cancer patients before therapy were significantly higher than those in healthy people（P < 0.05）. Conclusion：GC-MSC-CM-derived HGF plays an important role in promoting gastric cancer cells proliferation，migration，colony formation，PD-L1 expression and tumor formation，which may become apotential molecular target for gastric cancer prevention and treatment.

Abstract:Objective：This stady aims to observe the effects of continuous exposure to static magnetic field（SMF）on the proliferation and differentiation of dental pulp stem cells（DPSCs）. Methods：The magnetic cell culture set was used with the flux intensity about （35 ± 5）mT. The normal cultured cells were used as control. The morphology of the cells was detected by scanning electron microscopy（SEM）. The cell proliferation was detected by CCK-8，and the alkaline phosphatase activity（ALP）was also measured. Alizarin red staining（ARS） was used to detect the mineral synthesis. Results：There were no significant differences in SEM images and cell proliferation between magnetic cultured cells and normal cultured cells，but the ALP activity of magnetic cultured cells was significantly increased（P < 0.05），and the mineralization formation of cells under magnetic culture was significantly more than that of normal cultured cells（P < 0.05）. Conclusion：Continuous exposure to SMF neither changed the cell morphology nor the cell proliferation，but significantly promoted the differentiation of DPSCs.

Abstract:Objective：This study aims to observe the protective effect of periodontitis gene vaccine pVAX1-HA2-FimA and pVAX1-HA2-FimA/IL-15 in SD rats with experimental periodontitis. Methods：Fifty-four male SD rats were randomly divided into 9 groups：Control group（normal saline group，group A），empty plasmid pVAX1 group（group B），periodontal disease group（group C），Experimental group［50 μg，100 μg，150 μg pVAX1-HA2-FimA group（D，E，F group）and 50 μg，100 μg，150 μg pVAX1-HA2-FimA/IL-15 group（G，H，I group）］. Experimental periodontitis model was established after nasal drip immunization of SD rats，and the relative expression levels of TNF- α and MMP-8 mRNA in gingiva were detected by RT-qPCR. The absorption of alveolar bone was observed and measured by stereomicroscopy. Results：①The TNF-α and MMP-8 mRNA in the gingival tissues of each experimental group were lower than that in the periodontal disease group，and the differences were statistically significant（P < 0.05）. ②Compared with the pVAX1-HA2-FimA group，the amount of alveolar bone absorption in the pVAX1-HA2-FimA/IL-15 group with the same vaccine dose was lower，and the differences were statistically significant（P < 0.05）. Conclusion：The periodontitis gene vaccines pVAX1-HA2-FimA/IL-15 and pVAX1-HA2-FimA have protective effects on SD rats with experimental periodontitis.

Abstract:Objective：This study aims to analyze the expression of miR-1247 in human osteosarcoma cell lines，as well as its effect and mechanism on proliferation and apoptosis of osteosarcoma cells. Methods：qRT-PCR was performed to evaluate expression of miR-1247 in human osteosarcoma cell lines（U2OS，Saos-2 and 143B）and human normal osteoblast cell line（hFOB1.19）. Saos-2 cell line were divided into two groups，miR-1247 group transfected with miR-1247 mimics and EV group transfected with miR-1247 scramble. MTT assay and flow cytometry was used to assess proliferation and apoptosis ability of cells. Western blot was carried out to determine Sox9 and FAM129B proteins expression. Results：miR-1247 expression level in osteosarcoma cell lines was significantly lower than that in normal osteoblast cell line （P < 0.05）. There was no significantly difference in cell proliferation between EV and miR-1247 group after transfect for 0，1，3 d（P > 0.05）. The cell proliferation of miR-1247 group was significantly lower than that of EV group after transfect for 5，7 d（P < 0.05）. The apoptosis rate of miR-1247 group was significantly higher than that of EV group（P < 0.05）. The expression level of SOX9 and FAM129B protein of miR-1247 group was significantly lower than that of EV group（P < 0.05）. Conclusion：Osteosarcoma cell lines showed lower miR-1247 expression compare to normal osteoblast cell line. miR-1247 inhibit proliferation and induce apoptosis in osteosarcoma cell lines via down-regulation of Sox9 and FAM129B proteins.

Abstract:Objective：This study aims to investigate the endoplasmic reticulum correlative protein 1 （Derlin-1） expression in non-alcoholic fatty liver disease（NAFLD）and its effect on PERK（protein kinase R-like ER kinase）signal pathway to inhibit NAFLD. Methods：The expressin of Derlin-1 in liver tissues of NAFLD mice was detected by immuhistochemical staining and Western blotting. The over-expression of Derlin-1 in HepG2 cell was performed by plasmid vector transfection. Derlin-1 inhibits PERK pathway detected by Western blotting. Results：The expression of Derlin-1 in liver tissues of NAFLD mice was increased compared with that in nomal liver tissues. Overexression of Derlin-1 inhibited endoplasmic reticulum stress pathway activition. Conclusion：High expression of Derlin-1 in liver tissues of NAFLD pratients may play a critical role in decreasing endoplasmic reticulum stress and inhibiting liver cell pemelosis through inhibiting PERK expression.

Abstract:Objective：The aim of this study was to provide the reference for the diagnosis and differential diagnosis of related diseases，especially subacute thyroiditis by determining the general and regional distribution of the susceptible gene HLA-B*35 and its subtypes in Chinese mainland. Method：HLA high resolution typing was performed in 941 902 healthy adults in China. By classifing into 31 provinces，autonomous regions and municipalities，the overall and regional distribution characteristics of healthy adults and their HLA-B*35 genes and their subtypes in mainland China were determined. Results：The frequency of HLA-B *35 gene in healthy adult population in mainland China was higher in the northern population than that in the other regions. According to the classification of regions，the frequency of HLA-B *35 gene was the highest in Northwest China and was lower in South China. According to the calculation of provinces，municipalities and autonomous regions，the results showed that the gene frequency ranged from high to low in Inner Mongolia Autonomous Region（11.99%），Xinjiang Uygur Autonomous Region（11.76%），Ningxia Hui Autonomous Region（11.75%），Qinghai Province（11.61%）and Gansu Province（11.14%）. The lowest frequency was in Guangxi Zhuang Autonomous Region（3.79%）. The frequency of HLA-B* 3501 and HLA-B*3503 were the highest in Northwest China and the lowest in South China when classified by region. According to statistics of provinces and autonomous regions，the frequency of B*3501 is the highest in Inner Mongolia autonomous region and the lowest in Guangxi Zhuang autonomous region. Correspondingly，the frequency of B*3503 is the highest in Qinghai province and the lowest in Zhejiang province. Conclusion：The results showed that the distribution of HLA-B *35 in mainland China was high in the north and low in the south. Our study was conducted to explore the distribution of HLA-B*35 and its subtypes in different regions which is helpful for the diagnosis and differential diagnosis of related diseases.

Abstract:Objective：This study aims to evaluate the value of combined red blood cell distribution width（RDW）and neutrophil to lymphocyte ratio（NLR），along with model for end-stage liver disease（MELD） scores for predicting prognosis in patients with decompensated cirrhosis. Methods：A retrospective study was conducted on 181 patients with decompensated cirrhosis during January 2013 to December 2016. With death as the end point event，the patients were divided into death group and survival group according to the outcome of 1 year of follow-up. The differences of RDW，NLR and MELD levels at admission between the two groups were compared. Using multivariate Cox risk proportional model to predict independent predictors of the occurrence of 1-year mortality. The receiver operating characteristic（ROC） curve was drawn，and the area under the ROC curve（AUC） was compared. Kaplan-Meier survival analysis was carried out to compare the 1 year survival rate of patients with different RDW and NLR values. Finally，RDW，NLR and MELD scores were combined by binary logistic regression to assess the value of multivariate combination in predicting death within one year. Results：In 181 cases of decompensated cirrhosis，55 died and 126 survived within 1 years of follow-up. The value of NLR，RDW and MELD scores in the death group were statistically higher than those in the survival group（P＜0.05）. Multivariate Cox regression analysis suggested that NLR，RDW and MELD score were independent risk factors for predicting one-year mortality. The optimal cut-off values were 14.65% for RDW，2.58 for NLR，16.65 for MELD，with the AUC were 0.652，0.764 and 0.862，respectively. For combined measurements，the AUC was 0.870 for MELD score plus NLR and 0.876 for MELD score plus RDW，was increased to 0.884 for combination of RWD，NLR and MELD score. Conclusion：The combination of MELD，RDW and NLR has a higher predictive value for the 1-year prognosis of decompensated cirrhosis than MELD alone.

Abstract:Objective：This study aims to investigate the influencing factors of coronary slow flow（CSF）. Methods：The clinical data of 94 patients with CSF（CSF group）and 94 cases without CSF（control group）who underwent coronary angiography were recorded. The influencing factors of CSF and predictive value of platelet distribution width（PDW）for CSF were analyzed. Results：The ratio of diabetes of CSF group was higher than that in control group（P＜0.05）.The platelet（PLT），plateletocrit（PCT），mean platelet volume（MPV），platelet distribution width（PDW）and fasting plasma glucose（FPG）were different between the two groups（P＜0.05）. Multivariate logistic regression analysis showed that diabetes and PDW were the influencing factors of CSF（P＜0.05）. PDW in the CSF group increased significantly with increasing vessel involvement. ROC curve showed that the AUC of PDW predicting CSF was 0.675，optimum truncation value was 13.85%，sensitivity was 51.1% and specificity was 80.9%. Conclusion：Diabetes and PDW are the influencing factors of CSF and PDW may predict the occurrence of CSF.

Abstract:Objective：This study aims to explore the prognostic value of C-reactive protein-to-albumin（CRP/ALB）ratio（CAR）in diagnosed diffuse large B-cell lymphoma（DLBCL）. Methods：The clinical data of newly diagnosed DLBCL patients diagnosed in the First Affiliated Hospital of Nanjing Medical University between December 2009 and December 2018 were analyzed retrospectively. Progression-free survival （PFS） and overall survival （OS） were calculated by Kaplan-Meier method. Cox proportional hazards models were used for the univariate and multivariate analysis. Results：A total 314 newly diagnosed DLBCL patients were enrolled in the study. Up to June 2019，median follow up time was 50 months，and the median PFS and OS were unreached. Patients with higher CAR had shorter PFS and OS（P < 0.001）. The multivariate analysis strongly demonstrates that CAR is an independent predictor for both PFS（P=0.006）and OS（P=0.009）. Furthermore，the predictive and discriminatory capacity of National Comprehensive Cancer Network International Prognostic Index（NCCN-IPI）together with CAR level was superior to alone for PFS（P=0.010）and OS（P=0.002）. Conclusion：CAR is a simple and easily accessible parameter，which may be a good candidate for predicting prognosis of DLBCL.

Abstract:Objective：This study aims to evaluate the predictive value of serum HE4，CA125 and ROMA（Risk of Ovarian Malignancy Algorithm）in the diagnosis of ovarian malignancies. Methods：This research enrolled 370 patients with benign or malignant gynecological tumors treated in the First Affiliated Hospital of Nanjing Medical University. Serum levels of HE4 and CA125 were measured by electrochemiluminescence. The predicted probability of ovarian malignancy（PP） was calculated by ROMA method in 194 patients initially diagnosed as pelvic mass. The sensitivity，specificity，positive predictive value and negative predictive value of each indicators in diagnosing ovarian malignancy were calculated and the ROC curves were drawn. Results：The level of serum HE4 in ovarian malignancy group was significantly higher than that in other benign or malignant gynecological tumors groups（P < 0.05）. The level of serum CA125 in ovarian malignancy group was significantly higher than that in other groups except ovarian borderline tumors group and uterine adenomyosis group（P < 0.05）. The level of serum HE4 in patients with epithelial ovarian cancer（EOC）was significantly higher than that in patients without EOC（P < 0.05）. The levels of serum HE4 and CA125 in EOC patients with positive ascites were significantly higher than those in patients with negative ascites（P=0.001，P < 0.001）. The level of serum HE4 in EOC patients with abnormal D-dimer were significantly higher than those in patients with normal D-dimer（P=0.038）. The sensitivity of CA125 was significantly higher than that of HE4（P=0.031）. The positive predictive value of HE4 was significantly higher than that of ROMA or CA125（P < 0.05）. The ROC-AUC of ROMA was higher than that of CA125（P=0.006）. Conclusion：The specificity of serum HE4 was higher than that of CA125 in the diagnosis of ovarian malignancies. The combined ROMA index had better predictive value than detecting CA125 alone.

Abstract:Post-stroke anxiety（PSA）and post-stroke depression（PSD）are common sequelae after stroke，which not only destroys the quality of life of stroke patients，but also greatly interferes with their physical and mental health. There are many biological mechanisms and psychosocial mechanisms for the pathogenesis of post-stroke affective disorders，and different treatment strategies are generated by these different pathogenesis，including drug intervention and non-drug intervention. This article reviews the pathogenesis and clinical treatment of post-stroke affective disorder in recent years.

Abstract:Exosomes are secreted by cells，which has a nano-sized vesicular structure with a diameter of about 40-100 nm and is essentially a lipid bimolecular vesicle. Its main components and contents are mainly proteins，nucleic acids，and lipids. Not only in physiological but also in pathological conditions，exosomes can be secreted by most cells and exist in almost all body fluids，which is an important way of information exchange and material exchange. This review clarified the general characteristics of exosomes. The information exchange mode of exosomes in cells and the information exchange between central nervous cells were described in detail. The role of exosomes as molecular markers，particularly in neurodegenerative diseases，was explored. The drug treatment and gene therapy of exosomes in the central nervous system were briefly described.

Abstract:Interleukin-32（IL-32）is a novel proinflammatory cytokine，which is widely expressed in normal tissues. The expression level of IL32 is significantly increased in various cancer cells. IL-32 has been revealed to serve a crucial role in human cancer development，including tumor proliferation，migration，invasion and metastasis，which can be used as the biological marker of tumor prognosis. The studies on the molecular mechanisms underlying IL-32 are critical for the therapeutic strategies. This is a review of the current literature on the regulation function and the mechanisms of IL-32 in tumor invasion and immunity.