Abstract:Objective：This study aims to examine the expression of ZBTB3 in human glioblastoma(GBM)tissues and cell lines，and to explore the effects of ZBTB3 on the proliferation and clonal formation of GBM cells and their regulatory mechanism. Methods：The expression of ZBTB3 in tumor tissues of GBM patients was analyzed by GEPIA2 database. The mRNA and protein expression levels of ZBTB3 in GBM cell lines(U251，U373，U87)were detected by RT-PCR，qPCR and Western blot，and U87 cell line was identified with the highest expression of ZBTB3. CCK - 8 and clonal formation assay were used to examine the effects of silencing ZBTB3 on the proliferation and clonal formation of U87 cells. U87 cells were treated with p38MAPK，AMPK and Akt1 inhibitors，and the phosphorylation levels of p38MAPK，AMPK and Akt1 were detected by Western blot. ZBTB3 mRNA and protein levels were detected by RT - PCR，qPCR and Western blot. Cell proliferation and clone formation were examined by CCK - 8 and clone formation assay. Results：The expression of ZBTB3 in tumor tissues of GBM patients was significantly higher than that in normal tissues. The expression levels of ZBTB3 in U251，U373 and U87 cell lines were examined，and the highest expression in U87 cells was observed. Silencing ZBTB3 markedly inhibited the proliferation and clonal formation of U87 cells. AMPK inhibition could not only obviously reduce the expression level of ZBTB3 in U87 cells，but also markedly attenuate the proliferation and clonal formation of U87 cells. Conclusion：The expression of ZBTB3 is obviously increased in GBM tissues and cells，and the AMPK - up - regulated ZBTB3 expression promotes the proliferation and clonal formation of GBM cells.

Abstract:Objective：This study aims to explore the effect of overexpressing or silencing long intergenic non - protein coding RNA 01518(LINC01518)on the cell proliferation induced by IL -17 in non - small cell lung cancer(NSCLC). Methods：The IL -17 receptor A(IL-17RA)expression in NSCLC cell lines(PC9，H1299 and H1975)was first examined using Western blot. Then H1299 and PC9 cells were exposed to IL-17 for different time，and the cell proliferation was detected by CCK-8 assay. Based on the results of up-regulated lncRNAs detected by lncRNA microarray in IL-17-stimulated H1299 and PC9 cells，several lncRNAs were selected and verified by RT - PCR and Real - time PCR. Additionally，the constructed plasmids of pcDNA3.1/LINC01518 or shLINC01518 were transfected into H1299 cells，and H1299 cell proliferation were examined by CCK-8，EdU and colony formation assays. Results： IL-17RA could express in 3 kinds of NSCLC cell lines. The cell proliferation and LINC01518 were obviously up-regulated in H1299 and PC9 cells treated by IL-17. Besides，LINC01518 overexpression could markedly promote the cell proliferation，while LINC01518 gene knockdown followed by IL - 17 treatment did not enhance cell proliferation. Conclusion：LINC01518 can promote H1299 cell proliferation induced by IL-17 stimulation.

Abstract:Objective：This study aims to investigate the effects of serine/threonine protein kinase 25(STK25)on the activation of astrocytes as well as the process of experimental autoimmune encephalomyelitis(EAE)in mice. Methods：The EAE mouse model was induced by myelin oligodendrocyte glycoprotein 35-55(MOG_{35 - 55})polypeptide. The transcription levels and the expression of STK25， tumor necrosis factor- α(TNF-α)and interferon inducible protein 10(IP-10)in spinal cord tissue and serum were detected by ELISA， Real -time PCR and Western blot，respectively. The expression of STK25 in astrocytes in spinal cord tissue of mice was detected by immunofluorescence double staining. Primary mouse astrocytes were stimulated in vitro with interferon-γ(IFN-γ)，and the expression of STK25 and the transcription levels of cytokines were detected by the above methods at different times. Primary mouse astrocytes were transfected by recombinant lentiviral vector，and stimulated with IFN - γ，the transcription levels of TNF - α and IP - 10 were measured. Meantime，the EAE model was induced after injecting vectors 7 days，and the expression of STK25 in spinal cord tissue was detected by Western blot，the transcription levels of TNF - α and IP - 10 were detected by Real - time PCR，and the infiltration of inflammatory cells and the demyelination of spinal cord were observed by HE and Luxol fast blue(LFB)staining methods，respectively. Results：Compared with control group，the transcription level and the expression of STK25 in EAE group were significantly reduced while the levels of inflammatory cytokines and chemokines were significantly increased. Moreover，the expression of STK25 in astrocytes in spinal cord was significantly reduced. The expression of STK25 in astrocytes was decreased after stimulated by IFN-γ in vitro，and the production of inflammatory cytokines were increased. After specific knocking down endogenous STK25 in astrocyte，the production of inflammatory cytokines and chemokines in vitro astrocytes and in vivo EAE mice were increased significantly，which exacerbated the inflammatory cell infiltration and myelination of spinal cord tissue in EAE mice. Conclusion：STK25 may participate in the pathological process of EAE mice by affecting the activation of astrocytes and regulating the production of inflammatory cytokines and chemokines.

Abstract:Objective：The current study aims to explore the biological function of CCT2(chaperonin containning TCP1，subunit 2) in human lung adenocarcinoma. Methods：The expression divergence of CCT2 in lung adenocarcinoma tissues and normal lung tissue samples were analyzed from TCGA and GTEx database. The relationship between the expression level of CCT2 and the patient’s prognosis was evaluated through survival analysis. The expression of CCT2 in lung adenocarcinoma cells was measured by Reverse Transcription-Polymerase Chain Reaction(qRT-PCR)and Western blot. CCT2 siRNA was transfected into cell strains(A549，H1650 or H1299)respectively，which were divided into Group si - NC(negative control group)and Group si - CCT2(transfection group). Transfection efficiency was detected by qRT-PCR and Western blot. The proliferation ability of lung adenocarcinoma cell was tested by CCK-8. The cellular migration ability was detected by wound-healing experiment. The expression levels of CCT2 and Ki-67 in 72 cases of lung adenocarcinoma tissues were detected with immunohistochemical method，and the correlation between their expression levels and clinical pathology features were analyzed. Results：The results showed that，compared with normal lung tissues，the expression level of CCT2 in lung adenocarcinoma tissue was higher，which was related to low overall survival rate of lung adenocarcinoma patients (P＜0.05). According to qPCR and Western blot test，CCT2 highly expressed in lung adenocarcinoma cell，and the immunohistochemistry result showed that the CCT2 expression was positively correlated with the Ki-67 expression(P＜0.05). CCK-8 test results showed that the proliferation ability of A549 and H1650 cells in si-CCT2 group was significantly lower than that in the si- NC group 48 hours after siRNA transfection(P ＜ 0.05). The results of cell scratch test showed that the migration rate of A549 and H1650 cells in the si-CCT2 group was lower than that in the si-NC group 24 hours after siRNA transfection(P ＜ 0.05). Conclusion： The expression levels of CCT2 in lung adenocarcinoma cells and tissues increases. Knocking down its expression can inhibit the proliferation and migration of A549 and H1650 lung adenocarcinoma cells.

Abstract:Objective：This study aims to observe the effects of γ-Fe₂O₃-loaded chitosan porous sponge on the proliferation and early osteogenic differentiation of rat bone marrow mesenchymal stem cells(rBMSC). Methods：Chitosan sponges loaded with γ- Fe₂O₃ at concentrations of 1%，5%，10% and 20% were prepared by freeze -drying and cross -linking，and blank control group was prepared. Scanning electron microscopy and confocal microscopy were used to evaluate the adhesion and proliferation of rBMSC on the sponges. Cell proliferation at 1，3，5 and 7 days was detected by CCK -8. The early osteogenic differentiation of rBMSC at 7 and 14 days was evaluated by alkaline phosphatases(ALP)staining and activity detection. Real - time fluorescence quantitative reverse transcription PCR was used to detect the expression of ALP，bone morphogenetic protein 2(Bmp2)，collagen I(Col1)and Runt-related transcription factor 2(Runx2)after 7 and 14 days of osteogenic induction. The mineralization of extracellular matrix at 21 and 28 days was assessed by alizarin red staining quantitative method. Results：CCK-8 results showed each group added with γ-Fe₂O₃ promoted the proliferation of rBMSC. ALP staining and activity detection results showed the addition of γ-Fe₂O₃ can improve the activity of ALP. The results of real - time fluorescence quantitative reverse transcription PCR showed that the addition of γ - Fe₂O₃ can promote the expression of osteogenic indexes(ALP，Bmp2，Col1 and Runx2). The quantitative detection results of alizarin red staining showed that the amount of mineralization in the groups with γ-Fe₂O₃ added concentration of 5% and 10% was higher than that in the control group(P ＜ 0.05).Conclusion：The chitosan porous sponge loaded with γ - Fe₂O₃ promoted the proliferation and the early indicators of osteogenic differentiation of rBMSC，and loading γ-Fe2O3 at concentration of 5% and 10% can promote the formation of mineralization in the late stage of osteogenic differentiation of rBMSC.

Abstract:Objective：The current study aims to explore the effect of bergapten(BP)on the osteogenic differentiation of human dental pulp stem cells(hDPSCs). Methods：Specific antigens of hDPSCs were identified by flow cytometry analysis. The cytotoxicity and biosafety of BP was detected by CCK - 8. The hDPSCs were randomly divided into the control and BP groups. Different concentrations of BP(7.5 μmol/L，15.0 μmol/L and 30.0 μmol/L)were added to complete medium in the BP groups，while an equal volume of DMSO was added to the complete medium in the control group. The morphology of hDPSCs in the BP and control groups was observed by fluorescent stain. The BCIP/NBT staining and alkaline phosphatase(ALP)quantitative detection were used to detect the effect of BP at the early stage of osteogenic differentiation of hDPSCs. The alizarin red S(ARS)staining and semi-quantitative analysis was used to assess the degree of extracellular matrix mineralization at the late stage of osteogenic differentiation. Expressions of osteogenic-related genes such as Runt-related transcription factor 2(RUNX2)，osteocalcin(OCN)，osteopontin(OPN)，and ALP were detected by quantitative real -time PCR(qRT - PCR). Expressions of osteogenic - related proteins were detected using Western blot. Results：BP of 7.5 μmol/L，15.0 μmol/L and 30.0 μmol/L showed better biosafety(P ＜ 0.01). The ALP activities in the BP groups were higher than that in the control group，and the 15.0 μmol/L BP was the best(P ＜ 0.01). The degrees of extracellular matrix mineralization of the BP groups were higher than that of the control group，and the number of calcium nodules in the 15.0 μmol/L BP treatment group was more and deeper. The expressions of osteogenic-related genes and proteins were significantly increased in the BP groups and the 15.0 μmol/L BP treatment was the best(P ＜ 0.01). Conclusion：BP shows good biosafety and can promote the osteogenesis differentiation of hDPSCs with better effect at the 15.0 μmol/L concentration in vitro

Abstract:Objective：To compare the effects of different fixation methods on primary cilia immunofluorescence in mammalian cells and to investigate the optimized strategy of primary cilia immunofluorescence imaging. Methods：Cells were fixed with 4% paraformaldehyde(PFA)，methanol(MeOH)or 10% trichloroacetic acid(TCA)，followed by the same conditions of penetration， blocking and antibody incubation，and finally the primary cilia imaging results were compared under the same shooting parameters. Results：PFA fixation resulted in the better imaging of ciliary axoneme proteins and part of the ciliary membrane proteins，and MeOH fixation showed more clear imaging of the proteins localized at the bottom of cilia. Compared with other two methods，TCA fixation was suitable for some ciliary membrane proteins and axoneme binding proteins although the staining signal of ciliary axoneme was weak. In addition to the variations of fluorescence intensity，there were also minor differences in protein localization observed by staining after fixation using different methods. Conclusion：Each of the three fixation methods has their own advantages and limitations in immunofluorescent staining of primary cilia，and the appropriate fixation method needs to be selected with comprehensive consideration of the experimental purpose and the characteristics of target protein

Abstract:Objective：This study aims to explore the safety and feasibility of neoadjuvant chemotherapy combined with Da Vinci robotic surgery for advanced gastric cancer. Methods：The respective cross - sectional study was conducted. There were 151 patients retrospectively analyzed，who underwent radical gastrectomy after two cycles of neoadjuvant chemotherapy(SOX)in the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine from July 2018 to July 2022，and 120 cases were finally included according to the inclusion and exclusion criteria. According to mode of operation，all patients were divided into Da Vinci robotic radical gastrectomy group(robot group，60 cases)and laparoscopic radical gastrectomy group(laparoscopy group，60 cases). The general data， perioperative surgery related indexes and postoperative rehabilitation were compared between two groups. Results：There was no significant difference between the two groups in age，body mass index，tumor size，tumor location，surgical scope，digestive tract reconstruction mode，preoperative clinical stage(cTNM)，pathological stage(ypTNM)，cell differentiation degree，tumor regression grade，complete remission and partial remission(P ＞ 0.05). There were significant differences between the two groups in operation time，estimated intraoperative bleeding and abdominal incision length(P＜0.05). There were significant differences between the two groups in postoperative pain score，the time of first fluid diet，the time of first ventilation，the time of drainage tube extubation，the time of postoperative hospitalization and the cost of hospitalization(P ＜ 0.05). There were also significant differences in C-reactive protein， leukocyte count，neutrophil count，serum prealbumin，interleukin(IL)- 6 and procalcitonin between the two groups(P ＜ 0.05). However，there was no significant difference in the incidence rate of postoperative complications，readmission within 30 days after surgery，and secondary surgery within 30 days after surgery between the two groups(P ＞ 0.05). In the robot group，the estimated amount of surgical bleeding，the length of abdominal incision，the score of postoperative pain，the time of first liquid diet，the time of first ventilation，the time of extraction of abdominal drainage tube，and the time of postoperative hospitalization were better than those in the laparoscope group，but the operation time and hospitalization cost were significantly worse than those in the laparoscope group. Conclusion：For advanced gastric cancer，the strategy of preoperative neoadjuvant chemotherapy combined with Da Vinci robotic surgery is safe and feasible，and the incidence of postoperative complications is similar to laparoscopic surgery. So，Da Vinci robotic surgery system is a new candidate for advanced gastric cancer.

Abstract:Objective：The current study aims to establish a risk model for microvascular invasion(MVI)in patients with hepatocellular carcinoma(HCC)，which is based on radiological characteristics and clinical laboratory data，and to explore the clinical application of preoperative alpha -fetoprotein(AFP)and gamma -glutamyl transferase(GGT)in HCC patients. Methods：Clinical data from 479 HCC patients with surgically resection at the First Affiliated Hospital of Nanjing Medical University were retrospectively collected to investigate the effects of potentially relevant factors on MVI，and to assess the benefits of serum AFP and GGT on improving model predictive performance. Univariate and multivariate logistic regression analyses were used to select variables for the final prediction model，and the predictive performance of the model was assessed by area under the receiver operating characteristic curve(AUC)，calibration curve，and decision curve analysis(DCA). Predictive performance was compared by sensitivity，specificity and accuracy analyses. Kaplan - Meier curves were applied to assess the probability of patient survival. Results：In a multifactorial analysis，tumor size，borders of the tumor，exophytic growth，AFP and GGT were independent predictors for the development of MVI in HCC patients. Two models were constructed with or without AFP and GGT，respectively. In both cohorts，model A with AFP and GGT showed better predictive performance than model B without AFP and GGT，with AUCs of 0.902（0.872-0.932）and 0.876（0.842-0.909）， respectively. The two models showed good discrimination，and the predicted probabilities were in good agreement with the observed probabilities. Analysis of the DCA，which assesses the net clinical benefit of the models，showed that model A was slightly better than model B，both with good clinical application. In the subgroup analysis，AFP and GGT could well predict the prognosis of HCC patients. Conclusion：A predictive model for the development of MVI in patients with HCC was developed，and the study confirmed the clinical predictive value of serum AFP and GGT.

Abstract:Objective：Peripheral T -cell lymphomas(PTCL)is a heterogeneous group of non -Hodgkin’s lymphomas. The current study aims to analyze the clinical features of PTCL patients，and explore new factors that affected the progression-free survival(PFS) and overall survival(OS)，with which to establish new prognostic models. Methods：The clinical data of 202 patients with PTCL from the First Affiliated Hospital of Nanjing Medical University between July 2009 and September 2021 were retrospectively analyzed. Kaplan - Meier method，univariate and multivariate Cox regression analysis were performed for the survival analysis and prognostic factor evaluation. Results：The median PFS and OS of all PTCL patients were 11 months and 43 months respectively. Patients with lower level of hemoglobin(Hb)showed shorter PFS(9 months vs. 44 months，P ＜ 0.001)and OS(24 months vs. 83 months，P=0.002) than patients with normal level of Hb. The ECOG scores ＞1(P=0.009)and lower Hb level(P=0.007)were independent risk factors for PFS. Age ＞60 years(P=0.015)and the ECOG score ＞1(P=0.002)were independent risk factors for OS. International prognostic index (IPI)or prognostic index for T-cell lymphoma(PIT)combined with the Hb level improved the accuracy of predicting PFS of patients with PTCL. Conclusion：The level of Hb may be a good candidate for predicting prognosis of patients with PTCL. The Hb level combined with IPI and PIT can improve the ability to predict prognosis of patients with PTCL.

Abstract:Objective：The current study aims to explore the potential metabolic differences caused by systemic lupus erythematosus (SLE)and rheumatoid arthritis(RA)，the characteristics of serum metabolic profiles of patients with SLE and RA were analyzed. Methods：Serum samples were respectively collected from SLE patients，RA patients，and healthy volunteers. Hydrophilic interaction and ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(HILIC UHPLC-Q-TOF MS)was performed to analyze the full spectrum of these serum samples. The significantly altered metabolites were enriched by orthogonal partial least squares - discriminant analysis(OPLS -DA)models combined with univariate statistical analysis. The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic efficacy of the candidate serum metabolite biomarkers. KEGG pathway enrichment analysis was conducted to uncover the significantly altered pathways in each disease group. Results：The serum profiles showed remarkable differences among the SLE group，RA group，and the control group. Five significantly altered metabolites， including 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine，taurine，hexadecanedioic acid，(+-)12-hydroxyeicosantetraenoic acid(12 - HETE)and hypoxanthine，were proposed as potential diagnostic biomarkers. Furthermore，7 KEGG pathways，such as retrograde endocannabinoid signaling pathway，significantly altered in the SLE group when compared with the control group or RA group. Conclusion：Untargeted metabolomics study could be used to reveal the discrepancy of serum metabolic profiles among SLE patients，RA patients，and healthy people. The screened metabolite biomarkers and significantly altered pathways are expected to provide novel insights into the diagnosis and treatment of aforementioned diseases.

Abstract:Objective：This study aims to investigate the relevant factors of bone substitute materials retention rate after implant surgery with guided bone regeneration in the aesthetic zone. Methods：From January 2019 to December 2021，the cone beam CT data of 50 patients who received implant surgery in the aesthetic zone with guided bone regeneration in the implant department of the Affiliated Stomatological Hospital of Nanjing Medical University were collected before the operation，immediately after the operation and six months after the operation. Three-dimensional model reconstruction of the patient’s maxilla is performed in mimics19.0，along with registering three reconstructed models and calculating the retention rate of bone substitute materials，and its correlation with other clinical data was analyzed. Results：The retention rate of bone substitute materials in six months after the operation is statistically correlated with the use of concentrated growth factor，alveolar ridge splitting technique and thrombin time(P ＜ 0.05). There is a linear negative correlation between thrombin time and the retention rate of bone substitute materials(P ＜ 0.01). Conclusion：Both concentrated growth factor and alveolar ridge splitting technique are associated with the increase of bone substitute materials’retention rate in six months after the implant surgery，while long thrombin time is correlated with low bone substitute materials’retention rate.

Abstract:Objective：The current study aims to understand the characteristics and source apportionment of PM_2.5 from Jiangning university town of Nanjing during spring. Methods：Ambient PM_2.5 was collected from Jiangning campus of Nanjing Medical University between March 15^th and May 31^st in 2018. Thermal/optical carbon analyzer，ion chromatography，gas chromatography - mass spectrometer and energy dispersive X - ray fluorescence spectrometer were used to determine the concentration of carbonaceous components，water-soluble ions，polycyclic aromatic hydrocarbons(PAHs)and inorganic elements，respectively. Source apportionment was conducted by ion balance analysis，diagnostic ratio method，principal component analysis(PCA)，etc. Results：The average daily concentration of PM_2.5 was 70.24 μg/m3 . The ratios of c[organic carbon(OC)]/c[elemental carbon(EC)]mainly were 2.24~10.60， indicating fossil fuel combustion source and secondary pollution. Water soluble ions mainly included SO₄ ^2- ，NO₃ ^- and NH₄ ⁺ . The average ratio of c(NO₃ ^- )/c(SO₄ ^2- )was 0.65，indicating a coal combustion source. PAHs with 4~6 benzene rings were the most abundant. The results of PCA suggested a mixed coal，petroleum combustion and automobiles emissions source. Fe，Cl，K，Al and Zn were predominant in inorganic elements. Coal and petroleum combustion，automobiles emissions，industrial emissions as well as soil dust contributed to inorganic elements. Conclusion：PM_2.5 pollution is serious in the Jiangning university town of Nanjing. The main sources of PM_2.5 were the fossil fuel combustion and automobile emissions.

Abstract:Pericyte，a kind of mesenchymal cell，surrounds the wall of microvessels. Inflammation is the body’s defense response to tissue damage，which is helpful to the proliferation and repair of damaged tissues and the removal of harmful substances. However， excessive inflammatory reaction will also cause damage to the body and cause diseases. Studies in recent years have paid more and more attention to the role of pericytes in inflammation. Pericytes can quickly respond to inflammatory stimulation，regulate neutrophilic infiltration in process，and exert the interaction of endothelial cells through direct contact，paracrine and a series of signal transduction pathways，to maintain stable neurovascular unit and physiological barrier. In addition，pericytes themselves have the immune function of phagocytosis and antigen presentation，and have the dual role of anti - inflammatory and proinflammatory in the regulation of inflammation. These effects make pericytes play an important role in inflammatory diseases，and provide a possible research direction for the treatment of pericytes. This article will review how the pericytes function in inflammation and their role in inflammatory diseases，and review the research progress of related therapies.

Abstract:Gut dysbiosis has been identified as a potential factor that may drive a variety of diseases. Fecal microbiota transplantation(FMT)is a microecological therapy which delivers fecal microbiota of screened healthy donor into the recipient intestine. By restructuring the recipient gut microbiota with donor fecal samples，FMT has been used to treat a number of gastrointestinal and non - gastrointestinal disorders. Although FMT could cure 90% of recurrent Clostridioides difficile infections，the efficacy of FMT in other diseases is variable. These suggests that there are multiple factors influence the efficacy of FMT. This article will discuss the impact of both donor and recipient factors on donor microbiota engraftment and clinical efficacy in FMT.

Abstract:Environmental endocrine disrupting chemicals(EDC)are widespread in people’s production and living environment. Due to a wide variety of endocrine disruptors，they not only interfere with human reproductive system，nervous system and immune system，but also exert a non-negligible influence on the thyroid，which is the largest endocrine organ in the body. The previous studies mostly focused on thyroid hormone and histopathology，and few reports were available on the associations between autoimmune thyroid diseases(AITD)or thyroid cancer(TC)and EDC. Therefore，the current review elucidates the potential adverse effects of EDC on thyroid，especially in autoimmune thyroid diseases and thyroid cancer，including induction of thyroid autoantibodies and the effect on T cell differentiation，etc.，which may provide a basis for the study of the potential environmental exposure risk factors of autoimmune thyroid diseases and thyroid cancer，and has positive significance on prevention and treatment of these diseases.