Abstract:Objective：To study the binding of exogenous tumor necrosis factor receptor-associated factor 6(TRAF6)to Krüppel-like factors 5(KLF5)as well as the pattern and site of KLF5 polyubiquitination by TRAF6 in HEK 293T(i.e. 293 T)cells. Methods：The 293T cells were co-transfected with Flag-TRAF6，HA-KLF5 and ubiquitin(Ub)expression plasmids，or shTRAF6 and TRAF6 C70A plasmids in different combinations for 48 h. Then，the binding of TRAF6 to KLF5 and KLF5 K48/K63-linked polyubiquitination by TRAF6 were detected using immunoprecipitation(IP)and immunoblotting(IB)assays. Moreover，the plasmids with all lysine mutation of KLF5 were constructed，and co-transfected with TRAF6 overexpression plasmids into 293T cells. Thereafter，the level of KLF5 K63- linked polyubiquitination and the lysine(site)of KLF5 K63 -linked polyubiquitination were measured or identified by IP and IB. Results：TRAF6 and KLF5 in 293T cells could bind with each other. The overexpression of TRAF6 up - regulated while the knockdown or activity deficiency of TRAF6 down-regulated the level of KLF5 K63-linked polyubiquitination. The site of KLF5 K63- linked polyubiquitination was its K99 or K100 lysine. Conclusion：TRAF6 can interact with KLF5 and modify the K99 and K100 of KLF5 via K63-linked polyubiquitination.

Abstract:Objective：To investigate the effects of exogenous interleukin(IL)- 25 in gut damage in mice caused by Schistosoma japonicum(S.japonicum)infection. Methods：Twenty - four female C57BL/6J mice were randomly divided into four groups，control group(n=5)，control+IL-25(n=5)，S.japonicum infection group(Inf，n=7)，S.japonicum infection +IL-25 group(Inf+IL-25，n=7). Mice in the Inf group and Inf+IL-25 group were infected with forty S. japonicum cercariaes. IL-25 was intraperitoneally injected from the 4th week after infection(0.5 μg/mice every 2 days for 3 weeks). Six weeks after infection，mice were sacrificed. Hematoxylin eosin(HE) staining was performed in the liver and colon tissue to observed the pathological changes. Alcian blue-periodic and acid-Schiff(AB-PAS) staining was performed to observe the number of goblet cells in the colon. The levels of inflammation-related cytokines including IL-10， IL-4，IL-13，tumor necrosis factor α(TNF-α)，interferon γ(IFN-γ)，IL-1β in the colon were detected by enzyme linked immunosorbent assay(ELISA)and real - time polymerase chain reaction(real - time PCR). Results：HE staining results showed that the area of individual granulomas in the colon in the Inf+ IL -25 group was significantly lower than that of mice in the Inf group，but the area of granulomas in the liver in the Inf group was not significantly different from that in the Inf+IL-25 group. The results of AB-PAS staining showed that IL-25 could significantly promote the number of colonic goblet cells in mice after infection with S.japonicum，the ELISA and the real-time PCR results showed that cytokines in the colon after S.japonicum infection were obviously altered，and IL-25 injection promoted the expression of type 2 cytokines and decreased the expression of type 1 cytokines. Conclusion：IL-25 can alleviate the gut damage induced by S.japonicum infection by promoting the differentiation of goblet cells.

Abstract:Objective：To study the expression of C -X-C motif chemokine receptor 4(CXCR4)on regulatory T cells(Tregs)in peripheral blood(PB)and synovial fluid(SF)of rheumatoid arthritis(RA)and its correlation with clinical characteristics；so as to explore the pathogenesis of RA involving CXCR4 ⁺ Tregs. Methods：①The PB samples of 51 RA patients and 40 healthy volunteers were collected，and the knee SF samples of 10 patients with RA and 8 patients with osteoarthritis(OA)were collected by puncture of joint cavity. Then the ratio of CD4⁺ CD25⁺ CD127- Tregs and CXCR4⁺ Tregs in PB and SF was detected by flow cytometry. Clinical data of RA were collected，and the correlation analysis was made with Tregs and CXCR4 ⁺ Tregs ratio in PB. ②The peripheral blood mononuclear cells(PBMC)of RA and healthy volunteers were separated by Ficoll density gradient centrifugation method. CD4⁺ T cells were sorted by immunomagnetic positive selection kit and placed in the upper chamber of Transwell. C-X-C motif chemokine ligand 12(CXCL12)was added to the lower chamber. After 24 hours，the cells of the lower chamber were collected，and the CD4 ⁺ CD25 ⁺ CD127^- Tregs migrated to the lower chamber were detected by flow cytometry. Results：①Compared with healthy controls，the ratios of Tregs and CXCR4⁺ Tregs in PB of RA decreased，and the ratios in the high disease activity group and the middle disease activity group were lower than those in the remission group(P ＜ 0.05). The ratio of CXCR4 ⁺ Tregs in PB of RA was negatively correlated with the level of erythrocyte sedimentation rate(ESR)，C-reactive protein(CRP)，interleukin-6(IL-6)levels and DAS28 score. ②The ratios of Tregs and CXCR4⁺ Tregs in SF of RA were significantly higher than those in SF of OA. ③The ratios of Tregs and CXCR4⁺ Tregs in SF of RA were higher than those in PB. ④Compared with healthy controls，the mobility of Tregs in PB of RA increased significantly. Conclusion：The ratios of Tregs and CXCR4 ⁺ Tregs in PB of RA decreased，which are related to disease activity. Tregs and CXCR4 ⁺ Tregs in SF of joint inflammation are higher than those in PB and SF of OA. Tregs increased in SF of RA may be migrated from PB through CXCR4.

Abstract:Objective：To explore the neuroprotective effect of lovastatin(LOV)on N - methyl - D - aspartate(NMDA)induced excitotoxicity and the potential mechanism of LOV in regulating the function of NMDA receptors in neuroprotection. Methods：The primary cultured rat neurons were divided into the vehicle group，LOV group，NMDA group，LOV + NMDA group，glutamate(Glu) group and Glu + APV(a specific NMDA receptor antagonist)group. Neuronal morphology was detected by immunofluorescence staining，neuronal apoptosis was detected by TUNEL analysis，protein levels were detected by Western blotting，cell surface receptors were detected by biotinylation. Results：①Compared with the few surviving microtubule-associated protein 2(MAP-2)positive neurons in the NMDA group or the Glu group，the number of MAP -2 immunopositive neurons in the LOV+NMDA group and the Glu+APV group was significantly increased，as well as the number and length of neuronal dendrites were significantly increased(P ＜ 0.001). ②Compared with the significantly increased TUNEL-positive cells in the NMDA group or the Glu group，the TUNEL-positive cells in the LOV +NMDA group or the Glu +APV group were significantly decreased(P ＜ 0.001). ③Compared with the vehicle group，the expression of N - methyl - D - aspartate receptor(NR2B)in the NMDA group was significantly decreased(P ＜ 0.001)，while LOV pretreatment could increase the expression of NR2B when compared with the NMDA group(P ＜ 0.05). ④The cell surface receptor biotinylation assay showed that NMDA treatment resulted in the loss of the most NR2B on the cell surface(P ＜ 0.001)，while LOV pretreatment could significantly reduce the NMDA-induced loss of NR2B(P ＜ 0.05). Further studies showed that phosphorylation of NR2B at tyrosine(Tyr)1472 was decreased after NMDA treatment(P ＜ 0.05)，while pretreatment with LOV significantly restored the phosphorylation of Tyr1472(P ＜ 0.05). Conclusion：LOV may significantly attenuate the excitotoxicity induced by NMDA，and its neuroprotective effect is probably related to the selective regulation of NR2B surface expression by affecting the intracellular endocytosis and/or intracellular degradation of NR2B.

Abstract:Objective：To investigate the molecular mechanisms of gene expression and response in mouse kidney tissue during adaptation to hypoxic stress in the plateau based on transcriptome sequencing technology. Methods：C57BL/6 mice were reared at 4 200 m(high altitude kidney test group，HKT)and 400 m(plain kidney control group，PKC)above sea level，and kidney tissues were removed aseptically after 30 days for transcriptome sequencing using high-throughput RNA-sequencing(RNA-Seq). The sequencing results from the HKT and PKC groups were analyzed by gene ontology(GO)annotation and enrichment in the Kyoto encyclopedia of genes and genomes(KEGG)，and the reliability of the sequencing results was verified by real -time quantitative PCR(RT-qPCR). Results：Compared to the PKC group，1 349 genes expression were upregulated(P ＜ 0.05)and 1 658 genes expression were downregulated in the HKT group(P ＜ 0.05). Among them，prolactin receptor(PRLR)，apolipoprotein E(APOE)，apolipoprotein A4 (APOA4)，cytochrome C somatic(CYCS)，acyl-coenzyme A oxidase 1(ACOX2)，cytochrome C oxidase subunit 5A(COX5A)， cytochrome C oxidase subunit 5B(COX5B)，cytochrome C oxidase subunit 7A(COX7)and heat shock protein β-1(HSPB1)genes were significantly enriched. GO annotation analysis and KEGG enrichment analysis showed that differential genes were significantly enriched in organelle inner membrane，mitochondrial inner membrane and mitochondrial protein complexes，in addition to peroxisomal，oxidative phosphorylation，thermogenesis，carbon metabolism and tricarboxylic acid cycle pathways. Conclusion：Hypoxic stimulation at high altitude may cause oxidative stress，inflammatory response and imbalance of lipid metabolism in the body by affecting energy metabolism-related pathways.

Abstract:Objective：This study aims to explore the expression of ANKRD1 in hepatocellular carcinoma(HCC)and investigate the function and mechanism of ANKRD1 in promoting the proliferation and metastasis of HCC. Methods：The relative expression of ANKRD1 in hepatocellular carcinoma and adjacent tissues was detected by RT-qPCR and Western blot. Clinicopathological analysis was used to assess the association of ANKRD1 with the clinical feature of HCC patients. ANKRD1 was knocked down or overexpressed in HCC cell lines by lentiviral infection. The effects of ANKRD1 on the proliferation and migration of HCC cells were detected by CCK-8 assay，wound healing assay，transwell assay，EdU assay in vitro and subcutaneous tumor model in vivo. The activity of epithelial - mesenchymal transition(EMT)pathway was examined by Western blot. Results：ANKRD1 was upregulated in HCC. Overexpression of ANKRD1 promoted the proliferation，migration and invasion of HCC cells，while knockdown of ANKRD1 inhibited the proliferation and migration of HCC cells. Western blot and immunohisbochemical assay of subcutaneous tumor showed that ANKRD1 could activate EMT pathway. Conclusion：ANKRD1 is upregulated in HCC and promotes the proliferation and migration of HCC cells by regulating the EMT pathway，providing a potential therapeutic target for HCC.

Abstract:Objective：To investigate the regulatory mechanism of long non-coding RNA-nuclear enriched transcript 1(lnc-NEAT1) on the proliferation，migration and invasion of melanoma(MM)cells. Methods：MM cell lines(A375，A875，M14，B16)and human epidermal melanocytes(HEMa - LP)were cultured in vitro，real -time quantitative PCR(RT - qPCR)was performed to determine the mRNA expression of lnc-NEAT1，microRNA-29c-3p(miR-29c-3p)and chondroitin sulfate proteoglycan 4(CSPG4)in cells. B16 cells at logarithmic growth phase were taken and separated into control group，si-NC group，si-NEAT1 group，si-NEAT1+inhibitor-NC group， and si-NEAT1+miR-29c-3p inhibitor group. Lipofectamine 3000 was applied to transfect the corresponding plasmids into cells. RT-qPCR was performed to determine the transfection efficiency. MTT method was used to determine cell proliferation. Transwell assay was performed to determine cell migration and invasion abilities. Western blotting was performed to determine the expression of CSPG4 and proteins related to the proliferation，migration and invasion. Dual-luciferase reporter gene assay was performed to determine miR-29c-3p targeting relationship with lnc-NEAT1 and CSPG4. Nude mice xenograft experiment was performed to explore the effect of lnc-NEAT1 knockdown on the growth of MM cells in vivo. Results：The mRNA levels of lnc-NEAT1 and CSPG4 in MM cell lines were obviously higher than those in HEMa-LP cells，and the levels of miR-29c-3p were obviously lower than those in HEMa-LP cells(all P ＜ 0.05). Knockdown of lnc -NEAT1 obviously increased miR - 29c -3p expression，decreased CSPG4 mRNA and protein levels，inhibited cell proliferation，migration and invasion，and decreased Ki-67，N-cadherin and Vimentin protein levels as well as increased E-cadherin protein level(all P ＜ 0.05). Down - regulation of miR -29c -3p expression obviously increased CSPG4 mRNA and protein levels，and attenuated the inhibitory effects of lnc-NEAT1 knockdown on MM cell proliferation，migration and invasion(P ＜ 0.05). The results of dual-luciferase reporter gene assay showed that after the transfection of miR-29c-3p mimic，the luciferase activities of NEAT1-WT and CSPG4-WT in cells were obviously decreased(P ＜ 0.05). Nude mouse xenograft experiments showed that knockdown of lnc -NEAT1 obviously inhibited the growth of xenografts in vivo，while inhibition of miR-29c-3p was able to obviously attenuate the inhibitory effect of lnc-NEAT1 knockdown on the growth of xenografts(all P ＜ 0.05). Conclusion：Lnc-NEAT1 may promote the proliferation，migration and invasion of MM cells by regulating the miR-29c-3p/CSPG4 axis

Abstract:Objective：To investigate the effect of miR-27b-3p on osteosarcoma cells growth and its potential molecular mechanism. Methods：①The expression of miR-27b-3p in osteosarcoma cell lines was analyzed by RT-qPCR. ②CCK-8 assay，colony formation assay and flow cytometry analysis were used to detect the effect of miR-27b-3p and structure-specific recognition protein-1(SSRP1)on osteosarcoma cell growth. ③The miRNA target prediction database was used to predict potential target gene of miR-27b-3p，and dual luciferase reporter assay was performed to demonstrate the relationship of miR-27b-3p and its target genes. ④Efficiency of knockdown or overexpression was validated by Western blot analysis. Results：①The expression of miR -27b -3p in osteosarcoma cell lines and clinical samples was low，and overexpression of miR-27b-3p inhibited the growth of osteosarcoma cells. ②Bioinformatics analysis and dual-fluorescence reporter assay confirmed that SSRP1 was a target gene of miR-27b-3p，and the expression of SSRP1 was regulated by miR-27b-3p. ③Silencing of SSRP1 significantly inhibited cell proliferation and increased cell apoptosis of osteosarcoma cells. ④Overexpression of SSRP1 partly reversed the inhibitory effect of miR-27b-3p on osteosarcoma cell growth. Conclusion：miR-27b-3p is low expressed in osteosarcoma cells，and it regulates the growth of osteosarcoma cells by targeting SSRP1. Targeting the miR-27b-3p/ SSRP1 axis may become a new therapeutic strategy for the treatment of osteosarcoma.

Abstract:Objective：The aim of this study was to evaluate the associations between autonomic nervous function and constipation symptoms，psychological status and quality of life in function constipation(FC)patients. Methods：The characteristics，psychological status and heart rate variability(HRV)test of 102 FC patients and 42 healthy controls were evaluated. Then the FC patients were divided into three groups according to the result of HRV test，the autonomic tone balance normal group，the balance severely sympathetic tone group and the balance severely parasympathetic tone group. The results of self - rating anxiety and depression scale (SAS/SDS)，constipation scoring system(CSS)and patient assessment of constipation symptom(PAC - SYM)，patient assessment of constipation quality of life(PAC-QOL)were compared among three groups. Meanwhile，anorectal manometry and electrointestinogram parameters were assessed. We also analyzed the potential association of PAC - QOL with ANS function，mental status，constipation symptoms in all the constipation patients. Results：Compared with the healthy control group，the scores of SAS and SDS in FC patients increased significantly. HRV test results showed that the abnormal rates of autonomic nervous function were higher in FC patients than that in normal controls. Anorectal physiology and intestinal electric activity showed no significant difference among the FC patients with different HRV. Compared with patients in the autonomic tone balance normal group，patients in the balance severely sympathetic tone group had higher SAS/SDS scores，higher PAC - SYM score and lower PAC - QOL，especially for the dimension of“physical discomfort”and“worry/anxiety”of PAC -QOL. The balance severely parasympathetic tone group showed higher CSS score than the other two groups which maybe correlated with the prominence of depressed emotion. Multiple regression analysis showed that anxiety/ depression emotion and ANS function state could significantly affect the quality of life. Conclusion：FC patients with autonomic dysfunction are prone to anxiety or depression. The QOL of FC patients correlates strongly with ANS function and emotion，but not symptoms. Treatrnents such as autonomic adjustment，psychological interventions and central nervous system drugs may have clinical significance for FC patients.

Abstract:Objective：To explore the prognostic implication of the platelet - lymphocyte ratio(PLR)in primary central nervous system lymphoma(PCNSL)and to find a modified IELSG scoring system suitable for the Asian PCNSL population. Methods：The clinical data of 72 patients with PCNSL from the First Affiliated Hospital of Nanjing Medical University between June 2011 and December 2021 were retrospectively analyzed. Kaplan - Meier method，univariate and multivariate Cox regression analyses were performed for survival analysis and prognostic factors evaluation. Results：Median progression-free survival(PFS)and overall survival (OS)were 17 months and 39 months in PCNSL patients，respectively. Univariate Cox regression analysis confirmed PLR ≥107(P=0.022， P=0.038)，age 54≥ years(P=0.043，P=0.027)，LDH/ULN＞2(P=0.080，P=0.064)，CSF protein/ULN＞2(P=0.051，P=0.023)， ECOG≥2(P=0.029，P=0.027)and modified IELSG high - risk group(P=0.064，P=0.001)were associated with shorter PFS and OS. Multivariate Cox regression analysis confirmed PLR ≥107(P=0.011，P=0.022)and ECOG≥2(P=0.013，P=0.015)were independent risk factors for PFS and OS of PCNSL patients. Conclusion：PLR may be used as an indicator to evaluate the prognosis of PCNSL.

Abstract:Objective：To analyze the clinical features and prognosis of patients with anti-melanoma differentiation-associated gene 5(MDA5) positive dermatomyositis(DM) with pulmonary symptom as the first manifestation. Methods：The curren study retrospectively reviewed 57 patients with anti-MDA5 positive DM who were admitted to the First Affiliated Hospital of Nanjing Medical University between August 2017 and October 2021. The patients were divided into the pulmonary first symptom group(n=15)and the non - pulmonary first symptom group(n=42)according to the first clinical manifestation. The general clinical data，laboratory test results，imaging findings and prognosis were compared between the two groups. Results：There were statistically significant differences in therapeutic regimens between the two groups(P ＜ 0.05). The levels of serum ferritin(SF)and C - reactive protein(CRP)in the pulmonary first symptom group were significantly higher than those in the non-pulmonary first symptom group，and the level of creatine kinase(CK)was significantly lower than that in the non-pulmonary first symptom group(P ＜ 0.05). The main chest CT sign of patients in the pulmonary first symptom group was patchy shadow，followed by ground - glass opacity，thread net，and consolidation. Few interlobular septal thickening，nodular opacity，honeycomb lung，and pleural effusion were observed. Multivariate logistic regression analysis showed that the levels of SF and CK were independent risk factors for anti-MDA5 positive DM with pulmonary symptoms as the first manifestations(P ＜ 0.05). The cumulative survival rate within six months after the onset was significantly different in the two groups(P ＜ 0.05). Conclusion：Glucocorticoids combined with immunosuppressive agents is the main therapeutic regimen for anti - MDA5 positive DM patients with pulmonary symptoms as the first manifestations. The patients have poor prognosis，and their clinical manifestations and imaging findings are diverse. The levels of SF and CK are related to anti - MDA5 positive DM with pulmonary symptom as the first manifestation，which indicates that they can be used as laboratory monitoring indicators.

Abstract:Objective：To explore the diagnostic value of serum tenascin C(TNC)in bronchopulmonary dysplasia(BPD)of the premature infants. Methods：Seventy-nine very low birth weight premature infants were enrolled from Department of Neonatology，the Affiliated Huai’an No.1 People’s Hospital of Nanjing Medical University from 2018 to 2020. According to the 2018 NICHD diagnostic criteria，they were divided into the non -BPD group(n=48)and the BPD group(n=31). ELISA was used to detect serum TNC level； Multivariate logistic regression was conducted to analyze the independent risk factors of BPD；The correlation between serum TNC level and the severity of BPD was analyzed by Kendall correlation；Receiver operating characteristic(ROC)curve of TNC for diagnosis of BPD was made. Results：①The serum TNC level of preterm infants was statistically different between the non-BPD group and the BPD group[(95.03±19.73)ng/mL vs.(125.40±19.34)ng/mL，P ＜ 0.001]，and there was statistical difference between different levels of BPD groups(P ＜ 0.001). ②The independent risk factors for BPD in premature infants included birth weigh，high concentration oxygen inhalation time and serum TNC level(P ＜ 0.05). ③Serum TNC level was positively correlated with the severity of BPD(r =0.617， P ＜ 0.001). ④The area under the curve for the diagnosis of BPD by serum TNC was 0.881(P ＜ 0.001)，the sensitivity was 96.8%， and the specificity was 66.7%. Conclusion：Serum TNC level increases in BPD，and the diagnostic sensitivity and specificity is high， which can be used as one of the biomarkers for diagnosis of premature BPD.

Abstract:Objective：The current study aims to evaluate the correlation between inflammatory markers and 18F-fluorodeoxyglucose (FDG)positron emission tomography/computed tomography(PET/CT)metabolic parameters in patients with small cell lung cancer (SCLC)at baseline. Methods：A total of 56 SCLC patients，who underwent 18F - FDG PET/CT and did not receive any treatment in Department of Nuclear Medicine，the First Affiliated Hospital of Nanjing Medical University between January 2014 and December 2019，were analyzed retrospectively. The inflammatory markers[neutrophil/lymphocyte ratio(NLR)，platelet/lymphocyte ratio(PLR)， monocyte/lymphocyte ratio(MLR)and systemic immune-inflammation index(SII)]，clinical data and metabolic parameters of primary tumor[maximum standardized uptake value(SUVmax)，mean standardized uptake value(SUVmean)，metabolic tumor volume(MTV)，TLG(total lesion glycolysis)]within one week before PET/CT detection were collected to analyze the correlation by Spearman’s rho test. Results：The slight positive correlations were found between inflammatory markers(NLR，PLR，MLR，SII)and partial metabolic parameters[TLG(r_NLR=0.309，r_PLR=0.304，r_MLR=0.271，r_SII=0.362)，MTV(r_NLR=0.354，r_PLR=0.341，r_MLR=0.290，r_SII=0.411)]of primary tumor in 56 patients with SCLC(all P ＜ 0.05). The SUVmax and SUVmean were not found to be correlated with these hematological parameters(all P ＞ 0.05). The neutrophils，NLR，MLR，SII，MTV and TLG in extensive stage were higher than those in limited stage， and the lymphocytes in extensive stage were lower than those in limited stage(all P ＜ 0.05)，but SUVmax，SUVmean，PLR，monocytes and platelets were not found differences between extensive and limited stage(all P ＞ 0.05). Conclusion：Baseline NLR，MLR，SII of SCLC not only reflect the systemic metabolism，but also reflect the local inflammation of tumor. It may be necessary to receive whole- body PET/CT for staging to adjust treatment strategy in those patients with high NLR or MLR，SII.

Abstract:Objective：To construct a radiomic risk model to predict the risk of epileptic seizures in patients with diffuse low -grade glioma and to preliminarily assess patient outcomes. Methods：Using the imaging datasets and clinical data provided by TCGA and TCIA databases，an epilepsy risk score model was constructed by radiomics methods. Further nomograms were established based on the model combined with clinical indicators to predict the probability of epilepsy and the long-term survival rate. Receiver operator characteristic (ROC)curve and decision curve analysis were used to evaluate the predictive effectiveness of predictive models. Results：In the current study，10 radiomic features were filtered out to establish the epilepsy risk scoring model. The area under ROC curve of internal verification and external verification were 0.900 and 0.636，respectively. The positive rate of epilepsy in high - risk group and low - risk group were 80.6% and 27.3%，respectively. There was significant difference in 5-year overall survival between the high-risk and low-risk groups(P=0.029). Clinical decision curve analysis showed that the net benefit rate of the image-clinical model was higher than that of the clinical prediction model. The calibration curve showed that the 3-year，5-year survival nomogram had good calibration and differentiation ability and the 5-year overall survival between the high risk group and the low risk group in the nomogram had significant difference (P=0.008). Conclusion：In the current study，a radiomic model based on preoperative MRI of diffuse low-grade glioma was established for non-invasive prediction of the associated epilepsy and prognosis，which provides a reference for the individualized treatment strategies.

Abstract:Objective：To investigate the efficacy of Anlotinib combined with PD-1 inhibitor in the second-line treatment of elderly patients with advanced lung adenocarcinoma without driver gene mutation and its effect on T lymphocyte subsets. Methods：Elderly patients with advanced lung adenocarcinoma without driver gene mutation were given Pembrolizumab q3w combined with Anlotinib，or Nivolumab q2w combined with Anlotinib. The efficacy was evaluated by response evaluation criteria in solid tumors(RECIST)1.1 until the patient had disease progression or treatment intolerance. The changes of T lymphocyte subsets in peripheral blood before and after treatment were detected by flow cytometry，and the adverse reactions were monitored. Results：The objective remission rate was 28.1% and the disease control rate was 87.5%. The overall median progression free survival was 6.2 months. After two cycles of treatment，the levels of CD3⁺ ，CD4⁺ ，CD4⁺ /CD8⁺ were significantly higher than those before treatment，and the levels of CD8⁺ cells were significantly lower than those before treatment(P ＜ 0.01). Conclusion：In the second -line treatment of elderly patients with lung adenocarcinoma without driver gene mutation，Anlotinib combined with PD - 1 immunotherapy can improve the short - term efficacy，prolong the progression free survival，help the recovery of immune function and improve the quality of life，which has a good prospect in clinical application.

Abstract:Objective：To investigate the outcomes of non - obstructive azoospermia(NOA) patients with different etiologies undergoing microdissection testicular sperm extraction(micro - TESE). Methods：A retrospective analysis was carried out in 1 355 patients who accepted micro-TESE in Shanghai General Hospital from March 2015 to January 2022. The etiology/risk factors of NOA patients included Klinefelter syndrome(KS)，Y - chromosome AZFc deletion，cryptorchidism，mumps orchitis，chemoradiotherapy，varicocele，idiopathic etiology. The sperm retrieval rate(SRR)of patients in each group were analyzed，and the pregnancy outcomes of successful sperm retrieval in each group were compared. Results：The overall SRR was 26.2%(355/1 355). The mumps orchitis group ranked the highest SRR of 75.9%(22/29)，followed by cryptorchidism(70.5%，43/61)，Y-chromosome AZFc deletion(55.6%，30/54)， KS(47.6%，71/149)，idiopathic etiology(18.6%，167/897)，radiotherapy and chemotherapy(15.4%，2/13)，and varicocele was the lowest(13.2%，20/152). According to the clinical outcome，NOA patients were divided into a success group and a failure group. In idiopathic and chemoradiotherapy group，the levels of follicle-stimulating hormone(FSH)and luteinizing hormone(LH)in successful cases were significantly higher than those failed；In Y-chromosome AZFc deletion group，the levels of FSH was significantly lower than those failed；In mumps orchitis group，the volume of the testis in the success group was higher than those failed. In addition，we found that age can serve as an independent factor to predict sperm retrieval outcome in patients with idiopathic NOA，and the older had better sperm retrieval outcomes. After ICSI treatment，the pregnancy rate was 51.4%(200/389)，and the live birth rate was 73.5%(147/200). Conclusion：The SRR of NOA patients with different causes/risk factors are significantly different，which are important indicators affecting the outcome of micro-TESE.

Abstract:In 2022，multiple research milestones have occurred in the field of Alzheimer’s disease(AD). The genetic and pathological mechanisms of AD continued to be deepened and innovated. The rapeutics went through a fundamental transition from symptomatic modification therapy to targeted modification therapy，with amyloid-β targeted drugs gradually moving from clinical trials into real - world application. Encouraging findings on biomarkers and risk factors also marked the ongoing shift toward sprecise diagnosis and early prevention.

Abstract:With the aging process of society accelerating，the prevalence of Parkinson’s disease(PD)is rising. In recent years， plenty of evidence has proved that neuroinflammation plays a vital role in the progression of PD，especially around the risk areas in the basal ganglia and substantia nigra(SN). The damage and hyperactivation of microglia trigger oxidative stress，mitochondrial autophagy and autophagy dysfunction，the accumulation of α - synuclein and the release of pro - inflammatory cytokines，the neuronal damage mediated by these injury factors can further aggravate the activation of microglia，emphasizing the importance of this vicious cycle in the pathogenesis of PD. Therefore，exploring the role and mechanism of microglia in the pathogenesis of PD is of great significance for the diagnosis and treatment of PD.

Abstract:Stroke is a leading cause of adult disability worldwide，therefore promoting motor functional recovery from stroke is very important. Motor network remodeling is critical for motor functional recovery after stroke. The structural and functional reorganization of the motor network can be assessed by functional magnetic resonance imaging(fMRI). Environmental enrichment(EE)，which is a generally accepted strategy to promote stroke recovery，can be employed to reveal the molecular mechanisms underlying motor network remodeling. This paper reviewed the basal and clinical research progress of neural network remodeling after stroke.

Abstract:microRNA(miRNA)，a class of specialized RNA，which does not translate into proteins directly，but plays a role in the post - transcriptional level of genes，and it is one of the first non - coding RNAs to be studied. miRNA is richly expressed in the mammalian nervous system，and is involved in the regulation of neurodegenerative injury，stroke，brain trauma and other diseases. Recent studies have found that miRNA is especially associated with ischemic stroke. This paper summarized the latest progress of miRNA as biomarkers for diagnosis and prognosis of ischemic stroke and brain protection mechanisms as therapeutic targets，providing reference and basis for the clinical application of nucleic acid therapy with miRNA as the core.