Analysis of Significant Genes and Pathways in Esophageal Cancer Based on Gene Expression Omnibus Database

Objective To screen antigen targets for immunotherapy by analyzing over-expressed genes,and to identify significant pathways and molecular mechanisms in esophageal cancer by using bioinformatic methods such as enrichment analysis,protein-protein interaction(PPI)network,and survival analysis based on the Gene Expression Omnibus(GEO)database.Methods By screening with highly expressed genes,we mainly analyzed proteins MUC13 and EPCAM with transmembrane domain and antigen epitope from TMHMM and IEDB websites.Significant genes and pathways associated with the pathogenesis of esophageal cancer were identified using enrichment analysis,PPI network,and survival analysis.Several software and platforms including Prism 8,R language,Cytoscape,DAVID,STRING,and GEPIA platform were used in the search and/or figure creation.Results Genes MUC13 and EPCAM were over-expressed with several antigen epitopes in esophageal squamous cell carcinoma(ESCC)tissue.Enrichment analysis revealed that the process of keratinization was focused and a series of genes were related with the development of esophageal cancer.Four genes including ALDH3A1,C2,SLC6A1,and ZBTB7C were screened with significant P value of survival curve.Conclusions Genes MUC13 and EPCAM may be promising antigen targets or biomarkers for esophageal cancer.Keratinization may greatly impact the pathogenesis of esophageal cancer.Genes ALDH3A1,C2,SLC6A1,and ZBTB7C may play important roles in the development of esophageal cancer.

Methodological aspects of anti-human leukocyte antigen antibody analysis in solid organ transplantation

Donor human leukocyte antigen(HLA)-specific antibodies(DSA) play an important role in solid organ transplantation. Preexisting IgG isotype DSA are considered a risk factor for antibody mediated rejection, graft failure or graft loss. The post-transplant development of DSA depends on multiple factors including immunogenicity of mismatched antigens, HLA class Ⅱ typing of the recipient, cytokine gene polymorphisms, and cellular immunoregulatory mechanisms. De novo developed antibodies require special attention because not all DSA have equal clinical significance. Therefore, it is important for transplant clinicians and transplant immunologists to accurately characterize DSA. In this review, the contemporary immunological techniques for detection and characterization of anti-HLA antibodies and their pitfalls are described.

Identification of distant co-evolving residues in antigen 85C from Mycobacterium tuberculosis using statistical coupling analysis of the esterase family proteins

A fundamental goal in cellular signaling is to understand allosteric communication, the process by which sig-nals originating at one site in a protein propagate reliably to affect distant functional sites. The general principles of protein structure that underlie this process remain unknown. Statistical coupling analysis （SCA） is a statistical technique that uses evolutionary data of a protein family to measure correlation between distant functional sites and suggests allosteric communication. In proteins, very distant and small interactions between collections of amino acids provide the communication which can be important for signaling process. In this paper, we present the SCA of protein alignment of the esterase family （pfam ID： PF00756） containing the sequence of antigen 85C secreted by Mycobacterium tuberculosis to identify a subset of interacting residues. Clustering analysis of the pairwise correlation highlighted seven important residue positions in the esterase family alignments. These resi-dues were then mapped on the crystal structure of antigen 85C （PDB ID： 1DQZ）. The mapping revealed corre-lation between 3 distant residues （Asp38, Leu123 and Met125） and suggests allosteric communication between them. This information can be used for a new drug against this fatal disease.

Impact of human leukocyte antigen mismatching on outcomes of liver transplantation:A meta-analysis

AIM:To assess the effect of human leukocyte antigen(HLA) mismatching on liver graft outcome and acute rejection from a meta-analysis of available cohort studies.METHODS:Articles in PubMed/MEDLINE,EMBASE and the Cochrane database from January 1970 to June 2009,including non-English literature identified in these databases,were searched.Only studies comparing HLA or sub-phenotype matching with mismatching were extracted.The percentage of graft survival was extracted by "Engauge Digitizer" from survival curves if the raw data were not displayed.A meta-analysis was performed when at least 3 studies provided data.RESULTS:Sixteen studies met the inclusion criteria.A lower number of HLA mismatches(0-2 vs 3-6) did reduce the incidence of acute rejection(relative risk:0.77,P = 0.03).The degree of HLA mismatching(0-2 vs 3-6) had no significant effect on 1-year [hazard ratio(HR):1.04,P = 0.68] and 5-year(HR:1.09,P = 0.38) graft survival.In sub-phenotype analysis,the degree of HLA-A,B and DR mismatching(0 vs 1-2) had no significant effect on 1-year and 5-year graft survival,either.The HRs and P-values were 0.95,0.71(HLA-A,1-year);1.06,0.60(HLA-A,5-year);0.77,0.16(HLA-B,1-year);1.07,0.56(HLA-DR,1-year);1.18,0.23(HLADR,5-year),respectively.CONCLUSION:The results of this systematic review imply that good HLA compatibility can reduce the incidence of acute rejection in spite of having no influence on graft outcomes.To obtain a short recovery time and minimize rejection post transplantation,HLA matching studies should be considered before the operation.

Human leukocyte antigen class II DQB1*0301, DRB1*1101 alleles and spontaneous clearance of hepatitis C virus infection: A meta-analysis

AIM: To assess the associations of human leukocyte antigen (HLA) class Ⅱ DQB1*0301 and/or DRB1*1101 allele with spontaneous hepatitis C virus (HCV) clearance by meta-analysis of individual dataset from all studies published till date.METHODS: To clarify the impact of HLA class Ⅱ polymorphisms on viral clearance, we performed a metaanalysis of the published data from 11 studies comparing the frequencies of DQB1*0301 and DRB1*1101 alleles in individuals with spontaneous resolution to those with persistent infection. As we identified the heterogeneity between studies, summary statistical data were calculated based on a random-effect model.RESULTS: Meta-analyses yielded summary estimatesodds ratio (OR) of 2.36 [95%CI (1.62, 3.43), P<0.00001]and 2.02 [95%CI (1.56, 2.62), P<0.00001] for the effects of DQB1*0301 and DRB1*1101 alleles on spontaneous clearance of HCV, respectively.CONCLUSION: These results support the hypothesis that specific HLA class Ⅱ alleles might influence the susceptibility or resistance to persistent HCV infection.Both DQB1*0301 and DRB1*1101 are protective alleles and present HCV epitopes more effectively to CD4+T lymphocytes than others, and subjects with these two alleles are at a lower risk of developing chronic HCV infection. Large, multi-ethnic confirmatory and welldesigned studies are needed to determine the host genetic determinants of HCV infection.

Genetic testing vs microforceps biopsy in pancreatic cysts:Systematic review and meta-analysis

BACKGROUND Carcinoembryonic antigen(CEA)and cytology in pancreatic cystic fluid are suboptimal for evaluation of pancreatic cystic neoplasms.Genetic testing and microforceps biopsy are promising tools for pre-operative diagnostic improvement but comparative performance of both methods is unknown.AIM To compare the accuracy of genetic testing and microforceps biopsy in pancreatic cysts referred for surgery.METHODS We performed a literature search in Medline,Scopus,and Web of Science for studies evaluating genetic testing of cystic fluid and microforceps biopsy of pancreatic cysts,with endoscopic ultrasound with fine-needle aspiration(EUSFNA)prior to surgery and surgical pathology as reference standard for diagnosis.We evaluated the diagnostic accuracy for:1-benign cysts;2-mucinous low-risk cysts;3-high-risk cysts,and the diagnostic yield and rate of correctly identified cysts with microforceps biopsy and molecular analysis.We also assessed publication bias,heterogeneity,and study quality.RESULTS Eight studies,including 1206 patients,of which 203(17%)referred for surgery who met the inclusion criteria were analyzed in the systematic review,and seven studies were included in the meta-analysis.Genetic testing and microforceps biopsies were identical for diagnosis of benign cysts.Molecular analysis was superior for diagnosis of both low and high-risk mucinous cysts,with sensitivities of 0.89(95%CI:0.79-0.95)and 0.57(95%CI:0.42-0.71),specificities of 0.88(95%CI:0.75-0.95)and 0.88(95%CI:0.80-0.93)and AUC of 0.9555 and 0.92,respectively.The diagnostic yield was higher in microforceps biopsies than in genetic analysis(0.73 vs 0.54,respectively)but the rates of correctly identified cysts were identical(0.73 with 95%CI:0.62-0.82 vs 0.71 with 95%CI:0.49-0.86,respectively).CONCLUSION Genetic testing and microforceps biopsies are useful second tests,with identical results in benign pancreatic cysts.Genetic analysis performs better for low-and high-risk cysts but has lower diagnostic yield.

Identification of key pathways and gene expression in the activation of mast cells via calcium flux using bioinformatics analysis

Mast cells are the main effector cells in IgE-associated allergic disorders,and we have reported that mucosal mast cells(MMCs)play a more important role in the development of food allergy(FA).IgE with antigen or calcium ionophore stimulation can lead to the activation of MMCs via a calcium-dependent pathway.The purpose of the present study was to identify gene signatures with IgE/antigen(dinitrophenyl-bovine serum albumin,DNP-BSA)or calcium ionophore(A23187)on the activation of MMCs.Differentially expressed genes between the two types of samples were identified with microarray analysis.Gene ontology functional and pathway enrichment analyses of differentially expressed genes were performed using the database for annotation,visualization,and integrated discovery software.The results showed that IgE/antigen and A23187 could induce degranulation,increase vacuoles,and elevate the cytosolic calcium concentration in MMCs.Furthermore,GeneChip analysis showed that the same 134 mRNAs were altered with IgE/DNP-BSA and A23187,suggesting that DNP-BSA/IgE and A23187 affect the same signal pathway partly in degranulation.KEGG analysis showed that the data were enriched in NF-κB,TNF,MAPK,transcription factor activity,DNA binding,and nucleic acid binding,suggesting that activation of MMCs is a complex process.The results provide new insights on MMCs activation.

Specific HLA-DQB1 alleles associated with risk for development of hepatocellular carcinoma:A meta-analysis

AIM:To evaluate the association of human leukocyte antigen(HLA)-DQB1 alleles with hepatocellular carcinoma(HCC) through meta-analysis of published data.METHODS:Case-control studies on HLA-DQB1 allele association with HCC published up to January 2010 were included in the analyses.The odds ratios(ORs) of HLADQB1 allele distributions in HCC patients were analyzed and compared with healthy controls.The meta-analysis software REVMAN 5.0 was applied for investigating heterogeneity among individual studies and for summarizing all the studies.A meta-analysis was performed using fixed-effect or random-effect methods,depending on the absence or presence of significant heterogeneity.Seven case-control studies containing 398 cases and 594 controls were included in the final analysis.RESULTS:Among the five family alleles,two(DQB1*02 and DQB1*03) were found to be significantly associated with the risk of HCC.The combined OR for the association of DQB1*02 and DQB1*03 allele with the risk for HCC was 1.78(95% CI:1.05-3.03,P = 0.03) and 0.65(95% CI:0.48-0.89,P = 0.007),respectively.Among the 13 specific alleles,two(DQB1*0502 and DQB1*0602) were significantly associated with risk of HCC.The combined OR for the association of DQB1*0502 and DQB1*0602 allele with the risk for HCC was 1.82(95% CI:1.14-2.92,P = 0.01) and 0.58(95% CI:0.36-0.95,P = 0.03),respectively.No significant association was established for other HLA-DQB1 family alleles and specific alleles.CONCLUSION:Our results support the hypothesis that specific HLA-DQB1 allele families and alleles might influence the susceptibility or resistance to HCC,although it needs further investigations.

Effects of telbivudine and entecavir for HBeAg-positive chronic hepatitis B: A meta-analysis

AIM:To compare the effects of telbivudine (LDT) and entecavir (ETV) in treatment of hepatitis B e antigen (HBeAg)-positive chronic hepatitis B by meta-analysis. METHODS:We conducted a literature search using PubMed, MEDLINE, EMBASE, the China National Knowledge Infrastructure, the VIP database, the Wanfang database and the Cochrane Controlled Trial Register for all relevant articles published before April 1, 2012. Randomized controlled trials (RCTs) comparing LDT with ETV for treatment of HBeAg-positive chronic hepatitis B were included. The data was analyzed with Review Manager Software 5.0. We used relative risk (RR) as an effect measure, and reported its 95% CI. Meta-analysis was performed using either a fixedeffect or random-effect model, based on the absence or presence of significant heterogeneity. Two reviewers assessed the risk of bias and extracted data indepen- dently and in duplicate. The analysis was executed using the main outcome parameters including hepatitis B virus (HBV) DNA undetectability, alanine aminotransferase (ALT) normalization, HBeAg loss, HBeAg seroconversion, drug-resistance, and adverse reactions. Meta-analysis of the included trials and subgroup analyses were conducted to examine the association between pre-specified characteristics with the therapeutic effects of the two agents. RESULTS:Thirteen eligible trials (3925 patients in total) were included and evaluated for methodological quality and heterogeneity. In various treatment durations of 4 wk, 8 wk, 12 wk, 24 wk, 36 wk, 48 wk, 52 wk, 60 wk and 72 wk, the rates of HBV DNA undetectability and ALT normalization in the two groups were similar, without statistical significance. At 4 wk and 8 wk of the treatment, no statistical differences were found in the rate of HBeAg loss between the two groups, while the rate in the LDT group was higher than in the ETV group at 12 wk, 24 wk, 48 wk and 52 wk, respectively (RR 2.28, 95% CI 1.16, 7.03, P = 0.02; RR 1.45, 95% CI 1.16, 1.82, P = 0.001; RR 1.45, 95% CI 1.11, 1.89, P = 0.006; and RR 1.86, 95% CI 1.04, 3.32, P = 0.04). At 4 wk, 8 wk, 60 wk and 72 wk of the treatment, there were no significant differences in the rate of HBeAg seroconversion between the two groups, while at 12 wk, 24 wk, 48 wk and 52 wk, the rate in the LDT group was higher than in the ETV group (RR 2.10, 95% CI 1.36, 3.24, P = 0.0008; RR 1.71, 95% CI 1.29, 2.28, P = 0.0002; RR 1.86, 95% CI 1.36, 2.54, P < 0.0001; and RR 1.87, 95% CI 1.21, 2.90, P = 0.005). The rate of drug-resistance was higher in the LDT group than in the ETV group (RR 3.76, 95% CI 1.28, 11.01, P = 0.02). In addition, no severe adverse drug reactions were observed in the two groups. And the rate of increased creatine kinase in the LDT group was higher than in the ETV group (RR 5.58, 95% CI 2.22, 13.98, P = 0.0002). CONCLUSION:LDT and ETV have similar virological and biomedical responses, and both are safe and well tolerated. However, LDT has better serological response and higher drug-resistance.

ANALYSIS ON EPITOPES OF IGM WITH MONOCLONAL ANTI-ISOTYPIC AND ANTI-IDIOTYPIC ANTIBODIES AGAINST IgM FROM B-CLL

A double antibodies additivity ELISA test was employed to identify the epltopes which can be recognized by monoclonal antibodies (McAbs) against IgM from B chronic lymphocyte leukemia (B-CLL). The computer grouping programme analysis showed that 4 and- isotypic MaAbs could be divided into two groups and 10 anti- idiotype McAbs could be divided into four groups. The result was consistent with that of the indirect sandwich ELISA and inhibition ELISA test. It suggested that there were at least 6 distinct IgM epitopes which can react specifically with 14 McAbs. Our study indicated that the combination of the additivity ELISA test and the computer grouping programme analysis is of help in studying the relationship of the structure and function of antigen.

Development of Fok-I based nested polymerase chain reaction-restriction fragment length polymorphism analysis for detection of hepatitis B virus X region V5M mutation

AIM: To develop a Fok-I nested polymerase chain reaction(PCR)-restriction fragment length polymorphism analysis(PRA) method for the detection of hepatitis B virus X region(HBx) V5 M mutation.METHODS: Nested PCR was applied into DNAs from 198 chronic patients at 2 different stages [121 patients with hepatocellular carcinoma(HCC) and 77 carrier patients]. To identify V5 M mutants, digestion of nested PCR amplicons by the restriction enzyme Fok-I(GGA TGN9↓) was done. For size comparison, the enzymetreated products were analyzed by electrophoresis on 2.5% agarose gels, stained with ethidium bromide, and visualized on a UV transilluminator.RESULTS: The assay enabled the identification of 69 patients(sensitivity of 34.8%; 46 HCC patients and 23 carrier patients). Our data also showed that V5 M prevalence in HCC patients was significantly higher than in carrier patients(47.8%, 22/46 patients vs 0%, 0/23 patients, P < 0.001), suggesting that HBx Ag V5 M mutation may play a pivotal role in HCC generation in chronic patients with genotype C infections.CONCLUSION: The Fok-I nested PRA developed in this study is a reliable and cost-effective method to detect HBx Ag V5 M mutation in chronic patients with genotype C2 infection.

剪切波弹性成像检测尿道周围前列腺组织弹性模量与良性前列腺增生患者血清前列腺特异抗原的相关性研究

目的:分析尿道周围前列腺组织弹性模量与良性前列腺增生(BPH)患者血清前列腺特异抗原(PSA)的相关性。方法:采用便利抽样法收集2019年10月至2022年10月在合肥市第二人民医院接受治疗的200例BPH患者,所有患者均进行超声剪切波弹性成像与血清PSA检查,剪切波弹性成像测量尿道周围前列腺组织弹性模量取平均值,依据国际前列腺症状评分(IPSS)结果将其分为轻度组(96例)、中度组(59例)及重度组(45例),同时纳入本院前列腺检查人群中非前列腺病变的30名相关资料为健康对照组,分析比较患者尿道周围前列腺组织弹性模量与血清PSA的相关性。结果:统计学分析显示,各组受检者前列腺弹性模量值比较差异均有统计学意义(F=190.914,P<0.05)。与健康对照组相比,轻度组、中度组及重度组患者前列腺弹性模量值显著升高,差异有统计学意义(t=6.572、14.172、18.441,P<0.05);与轻度组相比,中度组、重度组患者前列腺弹性模量值显著升高(t=7.853、18.274,P<0.05);与中度组相比,重度组患者前列腺弹性模量值显著升高(t=11.371,P<0.05);不同程度的BPH患者间血清PSA差异均有统计学意义(F=126.143,P<0.05)。与健康对照组相比,轻度组、中度组及重度组患者血清PSA显著升高(t=10.694,14.368、13.804,P<0.001);与轻度组相比,中度组、重度组患者血清PSA显著升高(t=6.401、13.047,P<0.05);与中度组相比,重度组患者血清PSA显著升高(t=7.293,P<0.001);健康对照组血清PSA与尿道周围组织弹性模量无显著相关性(P>0.05)。轻度组、中度组及重度组患者血清PSA与尿道周围组织弹性模量均呈显著正相关(r=0.314、0.296、0.354,P<0.05)。结论:BPH患者血清PSA水平与尿道周围前列腺组织弹性模量值显著升高,二者存在正相关的关系。

PCNA在子宫内膜癌组织中的表达及临床意义

目的探讨增殖细胞核抗原(PCNA)基因在子宫内膜癌(UCEC)组织中的表达及与临床病理特征及潜在的生物学功能的关系。方法通过肿瘤基因组图谱(TCGA)数据库获取PCNA mRNA在UCEC中的表达数据及相关病理参数。收集2020年1月至2022年12月通过术后病理确诊的UCEC组织标本20例和正常内膜组织标本20例。免疫组化技术检测PCNA蛋白在上述组织中的表达。采用单因素Cox回归、多因素Cox回归分析PCNA基因在UCEC患者中的预后价值。利用TIMER数据库分析PCNA基因与免疫细胞浸润的关系。利用基因集富集分析(GSEA)预测PCNA表达的相关通路。利用STRING在线分析以PCNA为中心相关蛋白之间的相互作用。结果PCNA基因mRNA和编码蛋白在UCEC组织中表达水平上调(P<0.001),且具有诊断价值。免疫组化染色显示,PCNA蛋白在UCEC细胞核中高表达。Cox回归分析结果显示年龄、临床分期、主要治疗结果、病理分级、残留组织、组织学分级、肿瘤侵袭度、放射治疗可作为UCEC潜在的预后因素。PCNA表达水平与免疫浸润细胞的丰度相关。GSEA预测结果显示PCNA高表达富集于细胞周期、同源重组、DNA修复等通路。结论PCNA基因可能是UCEC潜在的诊断和预后标志物。

双靶点嵌合抗原受体-T细胞治疗复发难治多发性骨髓瘤患者疗效和安全性的Meta分析

背景嵌合抗原受体(CAR)-T细胞免疫疗法已在多发性骨髓瘤(MM)中取得较好的疗效,最常见的靶点为B细胞成熟抗原(BCMA)。单靶点CAR-T细胞免疫疗法的缺点是会导致疾病抵抗和复发,可能与抗原逃逸有关。为此,改进开发了双靶点CAR-T细胞治疗复发难治多发性骨髓瘤(RRMM),此方面尚缺乏系统的临床分析。目的对RRMM患者应用双靶点CAR-T细胞免疫疗法治疗的有效性及安全性进行Meta分析。方法计算机检索PubMed、Embase、Cochrane Library、Web of Science、中国知网、万方数据知识服务平台、维普网7个数据库中有关双靶点CAR-T细胞治疗RRMM的单组率研究,检索时限为建库至2023-02-06。由2名研究人员使用自制的数据表单来提取收集数据,并采用非随机对照试验方法学评价指标进行文献质量评价。采用R Studio软件进行数据分析。结果共纳入9篇文献,包括200例既往接受过多线治疗的RRMM患者。双靶点CAR-T细胞疗法根据不同靶点可分为4类:BCMA+CD19、BCMA+CD38,BCMA+跨膜剂与钙调节亲环素配体的相互作用者(TACI)、BCMA+人信号淋巴细胞激活分子家族成员7(CS1),其中BCMA+CD19靶点的研究较多。根据输注形式不同CAR-T细胞疗法可分为4类:双特异性CAR-T细胞、联合或序贯输注两种不同CAR-T细胞、双顺反子结构、共转导。Meta分析显示,双靶点CAR-T细胞治疗RRMM的总缓解率(ORR)为90.0%(95%CI=0.849~0.943),完全缓解率(CRR)为54.6%(95%CI=0.416~0.673),微小残留病(MRD)阴性率为75.6%(95%CI=0.489~0.952),髓外病变(EMD)总缓解率为55.1%(95%CI=0.234~0.851),最后一次随访时的复发率为29.7%(95%CI=0.141~0.454),最后一次随访时的生存率为75.6%(95%CI=0.554~0.915),3~4级细胞释放因子综合征(CRS)发生率为16.4%(95%CI=0.094~0.245),神经毒性(ICANS)发生率为4.0%(95%CI=0~0.120)。敏感性分析提示结果稳定。Egger's检验结果显示,ORR(P=0.03)及EMD总缓解率(P=0.02)提示存在一定的偏倚风险;CRR(P=0.53)、MRD阴性率(P=0.79)、最后一次随访时的复发率(P=0.71)、生存率(P=0.98)、3~4级CRS发生率(P=0.90)、ICANS发生率(P=0.30)提示不存在发表偏倚。结论双靶点CAR-T细胞免疫治疗RRMM显示出良好的疗效和安全性,未来需要多中心、大样本、更长随访期的研究来进一步评估其疗效和安全性。

微小扇头蜱microRNA文库建立及miR-275靶蛋白Vg-2的生物信息学分析

【目的】建立饱血状态雌性微小扇头蜱的microRNA文库,鉴定所含microRNA的种类,分析miR-275靶蛋白Vg-2的二、三级结构,检测其信号肽、跨膜结构域、磷酸化及糖基化位点,预测其优势抗原表位。【方法】以雌性、饱血状态的微小扇头蜱为研究对象,通过高通量测序技术建立其microRNA文库,利用软件miRDP及检索miRBase数据库,鉴定饱血状态雌性微小扇头蜱所含的microRNA种类;运用在线软件EXPASY、PRABI与SWISS-MODEL等分析Vg-2蛋白质理化性质,推断其二、三级结构;运用在线软件SignalP 5.0、TMHMM、NetPhos 3.1及NETCGlyc 1.0检测该蛋白的信号肽、跨膜结构域、磷酸化与糖基化位点;利用在线软件ABCpred Prediction、Scratch、IEDB和NetCTL预测Vg-2蛋白的B、T细胞优势抗原表位。【结果】获得成熟的microRNA 383个,其中67个为已被鉴定和注释的microRNA,316个为新发现的microRNA;微小扇头蜱的Vg-2基因序列全长5040 bp,共编码1679个氨基酸;Vg-2蛋白为亲水性酸蛋白,分子大小为189.89 kDa,理论等电点(pI)为6.08,其二级结构以无规则卷曲为主要结构成分,而其三级结构却以β-折叠为主要结构成分;Vg-2蛋白存在268个磷酸位点,20个糖基化位点,无信号肽和跨膜结构域,拥有58个B细胞优势抗原表位和8个T细胞优势抗原表位。【结论】在饱血状态雌性微小扇头蜱体内含有383个microRNA,受miR-275调控的Vg-2蛋白为结构不稳定的亲水性酸蛋白,具有良好的抗原性与免疫源性。

基于响应面分析的高抗原活性猪链球菌2型疫苗培养基的研制与效果评价研究

试验旨在研制具有高抗原活性的猪链球菌2型(Streptococcus suis type 2,SS2)疫苗培养基。研究以SS2 CVCC60615株为试验对象,通过单因素试验、Plackett-Burman试验、最陡爬坡试验和响应面分析得到SS2疫苗培养基。培养基发酵验证结果显示,SS2疫苗培养基菌液最大活菌数为1.09×10^(9) CFU/mL,是TSB培养基最大活菌数的2.41倍。测定SS2不同时间点的抗原活性发现,在SS2发酵培养过程中当活菌数达到最大值后的1~3 h内细菌的抗原活性最高。小鼠感染试验表明,SS2抗原活性最高时制备的灭活疫苗与商品化灭活疫苗相比具有更高的免疫球蛋白G(IgG)抗体效价和疫苗保护率。研究表明,采用研制得到的高抗原活性SS2疫苗培养基制备的灭活疫苗具有IgG抗体效价高、免疫保护力强的优点。

2015—2022年我国嵌合抗原受体T细胞临床试验注册信息分析

目的探讨中国嵌合抗原受体T细胞(CAR-T)疗法相关临床试验研究现状及未来趋势,为CAR-T疗法的开发提供参考。方法检索2015—2022年中国临床试验注册中心与国家药品监督管理局药品审评中心药物临床试验登记与信息公示平台登记注册的所有CAR-T疗法相关临床项目信息,采用文献计量学方法,统计注册项目的注册题目、注册时间、注册类型、研究疾病、靶点、上市申请临床试验信息、已上市CAR-T疗法等信息,分析CAR-T疗法临床试验的特点与现状。结果共检索到注册研究277项,主要集中在江苏省、上海市、广东省、湖北省,其中干预性研究163项,观察性研究114项,100例以上的大样本研究仅5项;资金来源主要为企业赞助(50.5%);主要适应证为血液系统恶性肿瘤(203/277,73.3%);单靶点研究中,白细胞分化抗原19(CD19)占43%(79/183),B细胞成熟抗原(BCMA)占9%(17/183);平台登记的项目27项,14个品种,仅2个为已上市品种,26个项目的申办单位为国内企业;Ⅰ或Ⅱ期试验研究占比较大,为96%(26/27);中国目前已上市3种CAR-T疗法,其中2个原研单位为国内企业。结论中国CAR-T研究主要集中在医疗资源发达地区,由中国企业为主导开展,并主要集中在治疗血液系统恶性肿瘤。目前多数研究仍处于临床试验阶段,由于注册时间短,离完成研究到申请上市还需要一定时间。

一株高病毒载量PCV2毒株的基因组特征及序列分析

【目的】了解广东省某猪群中猪圆环病毒2型(Porcine circovirus type 2,PCV2)流行毒株的遗传进化情况,丰富PCV2分子流行病学数据,为当地PCV2疫苗候选株的选用和研发提供参考。【方法】使用qPCR方法对疑似PCV2的样品进行检测,发现1株具有高病毒载量的PCV2毒株,命名为GD222858。通过PCR方法进行全基因组分子克隆及遗传进化分析。使用MegAlign软件将该毒株ORF1、ORF2基因编码的氨基酸序列与PCV2同亚型参考毒株进行比对,分析氨基酸序列的相似性;采用DNAStar预测该毒株的Cap蛋白二级结构及B细胞表位,并与4株疫苗株DBN-SX07-2(HM641752)、LG(HM038034)、SH(HM038027)、ZJ(AY686764)的Cap蛋白抗原指数进行比对分析。【结果】GD222858毒株基因组长度为1767 bp。遗传进化分析表明该毒株属于PCV2d亚型。与国内外82株参考毒株的核苷酸相似性为91.4%~99.6%,与越南毒株Han8(GenBank登录号:JQ181600)的亲缘关系最近。在ORF1编码的Rep蛋白处发现多个特异性突变位点F70Y、F77L、W202R、N256S;ORF2编码的Cap蛋白相对保守。Protean预测Cap蛋白的氨基酸第5~18、24~25、39~41、48~49、57~65、99、101、112~114、139~140、145~150、162~165、175~181、188~189、205~211、227~232位置处均可能存在潜在的B细胞表位。GD222858毒株的Cap蛋白抗原指数与4株疫苗株均有差异,在氨基酸45~57、124~132、223~233位置处抗原指数明显高于4株疫苗株,且与疫苗株HM038034差异最大。【结论】GD222858毒株感染猪群的原因可能是Rep蛋白多个位点发生特异性突变及疫苗株选用不当所致。

基于生信分析探索FOSL1对肝癌索拉非尼治疗抵抗的影响

目的:探究FOS样抗原1(FOS-like antigen 1,FOSL1)在肝癌组织中的表达及其对索拉非尼治疗抵抗的影响。方法:从公共数据库中获取数据进行WGCNA分析,对获取的差异表达基因(differentially expressed genes,DEGs)进行GO功能富集和KEGG通路富集分析。选取血管生成通路差异表达基因,采用STRING数据库建立PPI网络,使用MCODE插件提取排名前7位的关键基因。在人类蛋白质图谱网站对这7个关键基因在肝癌血管内皮细胞表达情况进行筛选。采用Western blot法检测人微血管内皮细胞-1(human microvascular endothelial cells-1,HMEC-1)及肝癌血管内皮细胞(tumor-derived endothelial cells,TEC)中FOSL1的表达,CCK-8法检测HMEC-1及TEC细胞对索拉非尼敏感性,免疫组化检测肿瘤组织及正常肝组织中FOSL1表达。结果:GO功能富集分析图和KEGG通路富集分析图显示“索拉非尼抵抗”和“索拉非尼不抵抗”患者DEGs主要富集在管腔形成和血管生成等通路,KEGG通路主要富集在Wnt信号通路和VEGF信号通路。WGCNA分析得到关键模块,通过对关键模块基因筛选,最终得到JUND、JUNB、IL17RC、IL17RA、IL17F、IL1B、FOSL17个关键基因,其中FOSL1在血管生成中发挥重要作用。通过人类蛋白质图谱网站,发现FOSL1在肝脏各类细胞中表达,但在内皮细胞中表达较高。Western blot检测显示,FOSL1在TEC细胞中的表达水平明显高于HMEC-1细胞。免疫组化染色发现,肿瘤组织中FOSL1高表达,正常肝组织中FOSL1表达水平较低。CCK-8法检测显示,HMEC-1细胞对索拉非尼较敏感,而TEC细胞对索拉非尼不敏感。结论:FOSL1在肝癌血管内皮细胞中高表达,可能与促进血管内皮细胞增殖及索拉非尼治疗抵抗相关。

激素敏感性前列腺癌患者中PSMA PET/CT衍生参数与循环肿瘤DNA特征之间的相关性分析

背景与目的:前列腺特异性膜抗原(prostate-specific membrane antigen,PSMA)正电子发射计算机体层显像(positron emission tomography/computed tomography,PET/CT)和循环肿瘤DNA(circulating tumor DNA,ctDNA)的检测结果都是激素敏感性前列腺癌(hormone-sensitive prostate cancer,HSPC)治疗决策的参考依据。本研究旨在分析HSPC患者中PSMA PET/CT衍生参数与ctDNA特征之间的相关性。方法:回顾性纳入于复旦大学附属肿瘤医院就诊且接受PSMA PET/CT和ctDNA测序的间隔≤2周、有完整病历记录的HSPC患者。排除存在除前列腺癌外的活动性恶性肿瘤,以及组织学特征支持纯神经内分泌癌或小细胞癌诊断的患者。本研究经复旦大学附属肿瘤医院伦理委员会批注(伦理编号:1909207-12)。采用Spearman相关系数评价PSMA PET/CT衍生参数最大标准摄取值(maximum standardized uptake value,SUVmax)、总肿瘤体积(total tumor volume,TTV)、病灶摄取总量(total lesion uptake,TLU)与ctDNA分数(ctDNA%)之间的相关性。结果:共纳入60例HSPC患者,TP53(3.3%)、BRCA2(3.3%)和ATM(3.3%)是最常见的突变基因。在相关性分析中,ctDNA%与SUVmax有显著相关性(Spearman’s rho=0.272,P=0.036);ctDNA%与TLU(Spearman’s rho=0.160,P=0.222)和TTV(Spearman’s rho=0.162,P=0.215)无显著相关性。结论:SUVmax与ctDNA%之间有显著相关性,提示与无PSMA阳性病灶和低PSMA摄取病灶的患者相比,存在高PSMA摄取病灶的患者接受联合靶向治疗的概率增加,本研究结果有望作为制订个体化治疗方案的参考。