Novel methylation gene panel in adjacent normal tissues predicts poor prognosis of colorectal cancer in Taiwan

BACKGROUND It is evident that current clinical criteria are suboptimal to accurately estimate patient prognosis.Studies have identified epigenetic aberrant changes as novel prognostic factors for colorectal cancer(CRC).AIM To estimate whether a methylation gene panel in different clinical stages can reflect a different prognosis.METHODS We enrolled 120 CRC patients from Tri-Service General Hospital in Taiwan and used the candidate gene approach to select six genes involved in carcinogenesis pathways.Patients were divided into two groups based on the methylation status of the six evaluated genes,namely,the<3 aberrancy group and≥3 aberrancy group.Various tumor stages were divided into two subgroups(local and advanced stages)on the basis of the pathological type of the following tissues:Tumor and adjacent normal tissues(matched normal).We assessed DNA methylation in tumors and adjacent normal tissues from CRC patients and analyzed the association between DNA methylation with different cancer stages and the prognostic outcome including time to progression(TTP)and overall survival.RESULTS We observed a significantly increasing trend of hazard ratio as the number of hypermethylated genes increased both in normal tissue and tumor tissue.The 5-year TTP survival curves showed a significant difference between the≥3 aberrancy group and the<3 aberrancy group.Compared with the<3 aberrancy group,a significantly shorter TTP was observed in the≥3 aberrancy group.We further analyzed the interaction between CRC prognosis and different cancer stages(local and advanced)according to the methylation status of the selected genes in both types of tissues.There was a significantly shorter 5-year TTP for tumors at advanced stages with the promoter methylation status of selected genes than for those with local stages.We found an interaction between cancer stages and the promoter methylation status of selected genes in both types of tissues.CONCLUSION Our data provide a significant association between the methylation markers in normal tissues with advanced stage and prognosis of CRC.We recommend using these novel markers to assist in clinical decision-making.

Expression of connective tissue growth factor in tumor tissues is an independent predictor of poor prognosis in patients with gastric cancer

AIM: To examine the expression of connective tissue growth factor (CTGF), also known as CCN2, in gastric carcinoma (GC), and the correlation between the expression of CTGF, clinicopathologic features and clinical outcomes of patients with GC. METHODS: One hundred and twenty-two GC patients were included in the present study. All patients were followed up for at least 5 years. Proteins of CTGF were detected using the Powervision two-step immunostaining method. RESULTS: Of the specimens from 122 GC patients analyzed for CTGF expression, 58 (58/122, 47.5%) had a high CTGF expression in cytoplasm of gastric carcinoma cells and 64 (64/122, 52.5%) had a low CTGF expression. Patients with a high CTGF expression showed a higher incidence of lymph node metastasis than those with a low CTGF expression (P = 0.032). Patients with a high CTGF expression had significantly lower 5-year survival rate than those with a low CTGF expression (27.6% vs 46.9%, P = 0.0178), especially those staging Ⅰ+Ⅱ+Ⅲ (35.7% vs 65.2%, P = 0.0027). CONCLUSION: GC patients with an elevated CTGF expression have more lymph node metastases and a shorter survival time. CTGF seems to be an independent prognostic factor for the successful differentiation of high-risk GC patients staging Ⅰ+Ⅱ+Ⅲ. Over-expression of CTGF in human GC cells results in an increased aggressive ability.

Serum HER2 as a predictive biomarker for tissue HER2 status and prognosis in patients with gastric cancer

AIM To investigate the association between serum human epidermal growth factor receptor 2(HER2) extracellular domain(ECD) and tissue HER2 status, and the prognostic value of serum HER2 ECD in patients with gastric cancer.METHODS A total of 239 patients with gastric cancer were enrolled from December 2012 to June 2013. Serum HER2 ECD was determined by chemiluminescent assay, and tissue HER2 status was evaluated by immunohistochemistry and fluorescence in situ hybridization assay. A receiver operating characteristic(ROC) curve was plotted to identify the optimal cut-off value for serum HER2 ECD assay for predicting survival in gastric cancer patients.RESULTS Serum HER2 ECD was significantly correlated with tissue HER2 status(P < 0.001), tumor size(P < 0.001), and intestinal type of gastric cancer(P =0.021). Serum HER2 ECD levels differed significantly between patients with HER2-positive tissue expression and those with HER2-negative tissue expression. ROC analysis yielded an area under the curve value of 0.79(95%CI: 0.71-0.87, P < 0.001), with a sensitivity and specificity of 0.54(95%CI: 0.37-0.70) and 0.93(95%CI: 0.88-0.96), respectively. With a cut-off value of 24.75 ng/m L, high serum HER2 ECD had a negative impact on overall survival of the patients(HR: 1.93, 95%CI: 1.32-4.38, P = 0.006). CONCLUSION Serum HER2 ECD could be a highly specific surrogate biomarker for tissue HER2 status in gastric cancer. Optimal cut-off criteria for predicting survival should be established.

Differential DNA methylation analysis of SUMF2,ADAMTS5,and PXDN provides novel insights into colorectal cancer prognosis prediction in Taiwan

BACKGROUND Patients with colorectal cancer(CRC)undergo surgery,as well as perioperative chemoradiation or adjuvant chemotherapy primarily based on the tumor–node–metastasis(TNM)cancer staging system.However,treatment responses and prognostic outcomes of patients within the same stage vary markedly.The potential use of novel biomarkers can improve prognostication and shared decision making before implementation into certain therapies.AIM To investigate whether SUMF2,ADAMTS5,and PXDN methylation status could be associated with CRC prognosis.METHODS We conducted a Taiwan region cohort study involving 208 patients with CRC recruited from TriService General Hospital and applied the candidate gene approach to identify three genes involved in oncogenesis pathways.A methylation-specific polymerase chain reaction(MS-PCR)and Epi TYPER DNA methylation analysis were employed to detect methylation status and to quantify the methylation level of candidate genes in tumor tissue and adjacent normal tissue from participants.We evaluated SUMF2,ADAMTS5,and PXDN methylation as predictors of prognosis,including recurrence-free survival(RFS),progression-free survival(PFS),and overall survival(OS),using a Cox regression model and Kaplan–Meier analysis.RESULTS We revealed various outcomes related to methylation and prognosis.Significantly shorter PFS and OS were associated with the CpG_3+CpG_7 hypermethylation of SUMF2 from tumor tissue compared with CpG_3+CpG_7 hypomethylation[hazard ratio(HR)=2.24,95%confidence interval(CI)=1.03-4.85 for PFS,HR=2.56 and 95%CI=1.08-6.04 for OS].By contrast,a significantly longer RFS was associated with CpG_2 and CpG_13 hypermethylation of ADAMTS5 from normal tissue compared with CpG_2 and CpG_13 hypomethylation[HR(95%CI)=0.15(0.03-0.71)for CpG_2 and 0.20(0.04-0.97)for CpG_13].The relationship between the methylation status of PXDN and the prognosis of CRC did not reach statistical significance.CONCLUSION Our study found that CpG_3+CpG_7 hypermethylation of SUMF2 from tumor tissue was associated with significantly shorter PFS and OS compared with CpG_3+CpG_7 hypomethylation.CpG_2 and CpG_13 hypermethylation of ADAMTS5 from normal tissue was associated with a significantly longer RFS compared with CpG_2 and CpG_13 hypomethylation.These methylationrelated biomarkers which have implications for CRC prognosis prediction may aid physicians in clinical decision-making.

Tissue inhibitor of metalloproteinase-3 expression affects clinicopathological features and prognosis of aflatoxin B1-related hepatocellular carcinoma

BACKGROUND The dysregulation of tissue inhibitor of metalloproteinase-3(TIMP3)was positively correlated with the progression of hepatocellular carcinoma(HCC).However,it is not clear whether TIMP3 expression is associated with the clinico-pathological features and prognosis of aflatoxin B1(AFB1)-related HCC(AHCC).A retrospective study,including 182 patients with AHCC,was conducted to explore the link between TIMP3 expression in cancerous tissues and the clinico-pathological characteristics and prognosis of AHCC.TIMP3 expression was detected by immunohistochemistry and its effects on the clinicopathological features and prognosis of AHCC were evaluated by Kaplan-Meier survival analysis and Cox regression survival analysis.Odds ratio,hazard ratio(HR),median overall survival time(MST),median tumor recurrence-free survival time(MRT),and corresponding 95%confidential interval(CI)was calculated to RESULTS Kaplan-Meier survival analysis showed that compared with high TIMP3 expression,low TIMP3 expression in tumor tissues significantly decreased the MST(36.00 mo vs 18.00 mo)and MRT(32.00 mo vs 16 mo)of patients with AHCC.Multivariate Cox regression survival analysis further proved that decreased expression of TIMP3 increased the risk of death(HR=2.85,95%CI:2.04-4.00)and tumor recurrence(HR=2.26,95%CI:1.57-3.26).Furthermore,decreased expression of TIMP3 protein in tissues with AHCC was significantly correlated with tumor clinicopatho-logical features,such as tumor size,tumor grade and stage,tumor microvessel density,and tumor blood invasion.Additionally,TIMP3 protein expression was also negatively associated with amount of AFB1-DNA adducts in tumor tissues.CONCLUSION These findings indicate that the dysregulation of TIMP3 expression is related to AHCC biological behaviors and affects tumor outcome,suggesting that TIMP3 may act as a prognostic biomarker for AHCC.

Mucosa-associated lymphoid tissue lymphoma of the trachea treated with radiotherapy:A case report

BACKGROUND Mucosa-associated lymphoid tissue(MALT)lymphoma originates in the marginal zone of lymphoid tissue.lung is one of the most frequent non-gastrointestinal organs involved,here known as bronchus-associated lymphoid tissue(BALT)lymphoma.BALT lymphoma of unknown etiology,and most patients are asymptomatic.The treatment of BALT lymphoma is controversial.CASE SUMMARY A 55-year-old man admitted to hospital had a three-month history of progressively coughing up yellow sputum,chest stuffiness,and shortness of breath.Fiberoptic bronchoscopy revealed mucosal visible beaded bumps 4 cm from the tracheal carina at 9 o'clock and 3 o'clock,the right main bronchus,and the right upper lobe bronchus.Biopsy specimens showed MALT lymphoma.Computed tomography virtual bronchoscopy(CTVB)showed uneven main bronchial wall thickening and multiple nodular protrusion.BALT lymphoma stage IE was diagnosed after a staging examination.We treated the patient with radiotherapy(RT)alone.A total dose of 30.6 Gy/17 f/25 d was given.The patient had no obvious adverse reactions during RT.The CTVB was repeated after RT and showed that the right side of the trachea was slightly thickened.CTVB was repeated 1.5 mo after RT and again showed that the right side of the trachea was slightly thickened.Annual CTVB showed no signs of recurrence.The patient now has no symptoms.CONCLUSION BALT lymphoma is an uncommon disease and shows good prognosis.The treatment of BALT lymphoma is controversial.In recent years,less invasive diagnostic and therapeutic approaches have been emerging.RT was effective and safe in our case.The use of CTVB could provide a noninvasive,repeatable,and accurate method in diagnosis and follow-up.

Nomogram model predicting the overall survival for patients with primary gastric mucosa-associated lymphoid tissue lymphoma

BACKGROUND Increasingly extranodal marginal B-cell lymphoma of mucosa-associated lymphoid tissue,known as mucosa-associated lymphoid tissue(MALT)lymphoma,is a type of non-Hodgkin’s lymphoma.The prognosis of primary gastric MALT(GML)patients can be affected by many factors.Clinical risk factors,including age,type of therapy,sex,stage and family hematologic malignancy history,also have significant effects on the development of the disease.The available data are mainly focused on epidemiology;in contrast,few studies have investigated the prognostic variables for overall survival(OS)in patients with primary GML.Based on the realities above,we searched a large amount of data on patients diagnosed with primary GML in the Surveillance,Epidemiology and End Results(SEER)database.The aim was to develop and verify a survival nomogram model that can predict the overall survival prognosis of primary GML by com-bining prognostic and determinant variables.AIM To create an effective survival nomogram for patients with primary gastric GML.METHODS All data of patients with primary GML from 2004 to 2015 were collected from the SEER database.The primary endpoint was OS.Based on the LASSO and COX regression,we created and further verified the accuracy and effectiveness of the survival nomogram model by the concordance index(C-index),calibration curve and timedependent receiver operating characteristic(td-ROC)curves.RESULTS A total of 2604 patients diagnosed with primary GML were selected for this study.A total of 1823 and 781 people were randomly distributed into the training and testing sets at a ratio of 7:3.The median follow-up of all patients was 71 mo,and the 3-and 5-year OS rates were 87.2%and 79.8%,respectively.Age,sex,race,Ann Arbor stage and radiation were independent risk factors for OS of primary GML(all P<0.05).The C-index values of the nomogram were 0.751(95%CI:0.729-0.773)and 0.718(95%CI:0.680-0.757)in the training and testing cohorts,respectively,showing the good discrimination ability of the nomogram model.Td-ROC curves and calibration plots also indicated satisfactory predictive power and good agreement of the model.Overall,the nomogram shows favorable performance in discriminating and predicting the OS of patients with primary GML.CONCLUSION A nomogram was developed and validated to have good survival predictive performance based on five clinical independent risk factors for OS for patients with primary GML.Nomograms are a low-cost and convenient clinical tool in assessing individualized prognosis and treatment for patients with primary GML.

Hepatitis gene chip in detecting HBV DNA, HCV RNA in serum and liver tissue samples of hepatitis patients

OBJECTIVE: To study the preparation of diagnostic gene chip for detecting hepatitis B virus (HBV) and hepatitis C virus (HCV) and its accuracy in detecting HBV DNA and HCV RNA in serum and liver tissues. METHODS: The probes, which depend on the conservative gene fragment of hepatitis virus, was designed, synthesized and spotted on the modified glass. The probes and some other control probes were assembled on the diagnostic microarray of hepatitis virus. The gene of hepatitis virus, purified from blood or tissue, was labeled with fluorescence and hybridized to the microarray. The hybridized microarry was scanned with microarray scanner and the diagnostic result was analyzed from the scanning data. Fourty patients with hepatitis B virus and 40 healthy people or 40 patients with hepatitis C virus were subjected to detection of HBV DNA and HCV RNA with the hepatitis virus gene chip by the double-blind method. Paraffin liver specimens obtained from 99 cases of posthepatitic cirrhosis were used to detect HBV DNA. The liver tissues and serum from 15 cases of chronic hepatitis B were used to detect HBV DNA. Simultaneously, HBsAg and HBcAg were detected in the serum by fluorescence microparticle quantitation, HBV DNA and HCV RNA in the serum by PCR, and HBcAg in liver tissues by immunocytochemistry or HBV DNA by in situ molecular hybridization. RESULTS: Chip detection of serum specimens showed that 30 patients were HBV DNA positive and 10 HBV DNA negative in the 40 patients with HBV positive, 25 patients were HCV RNA positive and 15 patients were HCV RNA negative in the 40 patients with HCV positive, and all were HBV and HCV negative in the 40 healthy people. In 15 patients with HBV marker positive who were subjected to liver biopsy, 15 patients were detected HBV DNA positive in serum by gene chip, 15 patients HBcAg positive in liver tissues by immunocytochemistry, 14 patients HBV DNA positive in liver tissues by in situ molecular hybridization, and 14 patients HBV DNA positive in liver tissues by gene chip. Paraffin liver tissues specimens from the 99 patients with posthepatitis B cirrhosis showed that 67 patients were detected HBcAg positive by immunocytochemistry, 53 patients HBV DNA positive by in situ molecular hybridization, and 46 patients HBV DNA positive by gene chip. In the 46 patients, 40 patients were detected HBV DNA and HBcAg positive by in situ molecular hybridization and immunocytochemistry, 6 patients only HBcAg positive, and 33 patients HBcAg negative. CONCLUSIONS: The designed diagnostic gene chip can be used to simultaneously detect serum HBV DNA and HCV RNA, but the positive rate of HCV RNA diagnosed by this chip is lower. The gene chip can detect HBV DNA in serum and in liver tissue.

儿童胃肠道炎性肌纤维母细胞瘤的诊断与治疗

目的总结儿童胃肠道炎性肌纤维母细胞瘤(inflammatory myofibroblastic tumor,IMT)的临床特征及诊治经验。方法回顾性分析湖南省儿童医院普外科2010年1月至2021年12月收治的11例胃肠道IMT患儿临床资料,男7例,女4例;发病年龄8个月至15岁,收集患儿临床特点、影像学检查、病理学诊断、外科治疗及随访情况等。结果11例主要以腹痛、呕吐、发热、血便及腹部肿物就诊。10例行一期手术完整切除肿瘤,其中1例术后予化疗;1例经活检确诊后未予手术,仅行化疗。11例均病理诊断明确,均获随访(随访时间6~60个月),其中9例治愈,2例带瘤生存(1例于术后2年复发)。结论儿童胃肠道IMT临床少见,手术是首选治疗方法。复发、难治性IMT的治疗亟待进一步积累经验。

基于GEO数据库相关基因检测数据估算分析直肠癌组织免疫细胞分布变化

目的 基于GEO数据库相关基因检测数据估算直肠癌组织免疫细胞比例,分析不同免疫细胞之间以及免疫细胞与患者临床病理特征、预后的关系,以探讨直肠癌组织中免疫细胞的分布情况及其变化的临床意义。方法从GEO数据库中下载GSE87211数据集,从中提取直肠癌组织与直肠正常对照组织(简称对照组织)基因检测数据及直肠癌患者临床相关数据。应用CIBERSORT反卷积法估算直肠癌组织及直肠对照组织中22种免疫细胞(包括浆细胞、初始B细胞、记忆性B细胞、滤泡辅助性T细胞、调节性T细胞、γδT细胞、CD8+T细胞、初始CD4+T细胞、未活化CD4记忆T细胞、活化CD4记忆T细胞、未活化NK细胞、未活化肥大细胞、活化肥大细胞、嗜酸性粒细胞、中性粒细胞、活化NK细胞、单核细胞、M0巨噬细胞、M1巨噬细胞、M2巨噬细胞、未活化树突状细胞及活化树突状细胞)比例。采用Pearson相关性分析不同免疫细胞之间的相关性;应用Wilcox检验或Kruskal-Wallis检验比较不同临床病理特征直肠癌患者癌组织中免疫细胞比例;以不同免疫细胞比例中位值为界,将直肠癌患者分为高比例组和低比例组,采用Kaplan-Meier曲线对高、低比例组患者进行生存分析,并比较两组生存率。结果 与对照组织比较,直肠癌患者癌组织中初始CD4+T细胞、活化CD4记忆T细胞、调节性T细胞、未活化NK细胞、单核细胞、M0巨噬细胞、未活化树突状细胞、活化肥大细胞、中性粒细胞比例增加,记忆性B细胞、CD8+T细胞、浆细胞、未活化CD4记忆T细胞、滤泡辅助性T细胞、M2巨噬细胞、未活化肥大细胞比例降低(P均<0.05)。直肠癌组织中活化肥大细胞和中性粒细胞比例呈正相关(r=0.71,P<0.05),M2巨噬细胞和浆细胞细胞比例呈正相关(r=0.65,P<0.05),CD8+T细胞和浆细胞细胞比例呈正相关(r=0.62,P<0.05),中性粒细胞和M2巨噬细胞比例呈负相关(r=-0.62,P<0.05)。与KRAS野生型直肠癌患者相比,KRAS突变型直肠癌患者癌组织中活化CD4记忆T细胞及中性粒细胞比例增高、调节性T细胞比例降低(P均<0.05)。有淋巴结转移的直肠癌患者癌组织中浆细胞、γδT细胞及调节性T细胞比例低于无淋巴结转移患者。存在远处转移的直肠癌患者组织中未活化NK细胞比例高于未转移的患者(P均<0.05)。浆细胞高比例组的直肠癌患者较低比例组的患者具有更好的预后,中性粒细胞高比例组患者生存期短于低比例组(P均<0.05)。结论 直肠癌组织中免疫细胞分布与对照组织相比存在差异。不同临床病理特征直肠癌患者中免疫细胞亚群比例亦有所不同。浆细胞高表达、中性粒细胞低表达预示着患者更好的生存结局。

A Review of Type 1 and Type 2 Intraductal Papillary Neoplasms of the Bile Duct

Intraductal papillary neoplasm of the bile duct(IPNB)is a heterogeneous disease similar to intraductal papillary mucinous neoplasm of the pancreas.These lesions have been recognized as one of the three major precancerous lesions in the biliary tract since 2010.In 2018,Japanese and Korean pathologists reached a consensus,classifying IPNBs into type l and type 2 IPNBs.IPNBs are more prevalent in male patients in East Asia and are closely related to diseases such as cholelithiasis and schistosomiasis.From a molecular genetic perspective,IPNBs exhibit early genetic variations,and different molecular pathways may be involved in the tumorigenesis of type 1 and type 2 IPNBs.The histological subtypes of IPNBs include gastric,intestinal,pancreaticobiliary,or oncocytic subtypes,but type 1 IPNBs typically exhibit more regular and well-organized histological features than type 2 IPNBs and are more commonly found in the intrahepatic bile ducts with abundant mucin.Due to the rarity of these lesions and the absence of specific clinical and laboratory features,imaging is crucial for the preoperative diagnosis of IPNB,with local bile duct dilation and growth along the bile ducts being the main imaging features.Surgical resection remains the optimal treatment for IPNBs,but negative bile duct margins and the removal of lymph nodes in the hepatic hilum significantly improve the postoperative survival rates for patients with IPNBs.

儿童非横纹肌肉瘤类软组织肉瘤的临床特点及预后

目的 分析儿童非横纹肌肉瘤类软组织肉瘤(NRSTS)的临床特点、治疗及预后影响因素。方法 回顾性分析2012年8月—2022年8月确诊并在我院接受规范治疗的NRSTS患儿的临床资料,通过Kaplan-Meier法分析患儿的预后及影响因素。结果 共纳入49例患儿,男27例,女22例,中位诊断年龄52.7 (0.8~149.6)个月。病理亚型有14种,包括婴儿型纤维肉瘤9例(18.4%)、肾外横纹肌样瘤8例(16.3%)、炎性肌纤维母细胞瘤7例(14.3%)、肝未分化胚胎性肉瘤7例(14.3%)、其他病理亚型18例(36.7%)。在多学科综合治疗下,初治完全缓解43例,部分缓解1例,有效率89.8%。复发15例(30.6%),中位复发时间14 (3~40)个月,复发后中位生存时间16个月。中位随访时间37(4~125个月),3年OS为(74.9±6.6)%,3年EFS为(59.4±7.3)%。Ⅰ+Ⅱ期、Ⅲ+Ⅳ期的3年OS分别为(94.4±5.4)%vs(56.9±10.4)%(P=0.003),3年EFS分别为(72.9±9.6)%vs(46.2±10.3)%(P=0.042)。COG危险度分组低危组和中高危组3年OS分别为100%vs(62.0±9.2)%(P=0.009)。结论 儿童NRSTS治疗以多学科综合治疗为主,完全手术切除是预后良好的关键,COG中高危组预后不佳,复发患儿预后较差。

Correlations of hypoxia-inducible factor-1α/hypoxia-inducible factor -2α expression with angiogenesis factors expression and prognosis in non-small cell lung cancer

Background Hypoxia-inducible factor （HIF） may play an important role in the process of tumorigenesis as well as tumor progression. The aim of this study was to compare the expression between HIF-1α and HIF-2α in tumor angiogenesis and the overall impact on patient prognosis in human non-small cell lung cancer （NSCLC）. Methods In the current work we compared the immunohistochemical expression of HIF-1α and HIF-21 in surgical specimens of 140 patients with NSCLC in a tissue microarray study. Relationships between HIF-α expression and clinicopathological or angiogenic factors, including prognosis, were analyzed. Results High HIF-1α and HIF-2α expression was noted in 49/140 （35.0%） and in 64/140 （45.7%） of the cases, respectively. There was no direct correlation between HIF-la and HIF-2α expression. Patients with advanced stage tumors had frequent high expression of HIF-2a （P=0.007）, and we also found a significant correlation between HIF-2α and T or N stage （P=0.030 and 0.043, respectively）. HIF-1α showed a marginal association with T stage （P=0.084）, which showed a higher expression in early stage tumors. A significant correlation （p=0.045） was noticed between HIF-1α and vascular endothelial growth factor （VEGF） expression while the expression levels of thymidine phosphorylase （TP）, cyclooxygenase （COX）-2 and microvessel density （MVD） were significantly higher in high HIF-2a tumors （P=0.020, 0.004 and 0.046, respectively）. In addition, univariate analysis of overall survival demonstrated that HIF-2a expression, but not HIF-la, was related to poor outcome （P=-0.001） and it retained significant in multivariate analysis （P=0.036）. Conclusions Taken together, we conclude that HIF-1α and HIF-2α may differentially regulate the major angiogenic factors in different stages of the tumor process in NSCLC. HIF-2α may play a dominant role in tumor angiogenesis and appears to be of obvious value as a significant prognostic factor in NSCLC.

LAG-3 expression on tumor-infiltrating T cells in soft tissue sarcoma correlates with poor survival

Objective: To elucidate the role and prognostic significance of lymphocyte activation-gene-3(LAG-3) in soft tissue sarcoma(STS).Methods: The expression of LAG-3 in patient and matched normal blood samples was analyzed by flow cytometry. The localization and prognostic values of LAG-3^+ cells in 163 STS patients were analyzed by immunohistochemistry. In addition, the expression of tumor-infiltrating CD3^+ T, CD4^+ T, and CD8^+ T cells and their role in the prognosis of STS were evaluated by immunohistochemistry. The effect of LAG-3 blockade was evaluated in an immunocompetent MCA205 fibrosarcoma mouse model.Results: Peripheral CD8^+ and CD4^+ T cells from STS patients expressed higher levels of LAG-3 than those from healthy donors.LAG-3 expression in STS was significantly associated with a poor clinical outcome(P = 0.038) and was correlated with high pathological grade(P < 0.001), advanced tumor stage(P = 0.016). Additionally, LAG-3 expression was highly correlated with CD8^+ T-cell infiltration(r = 0.7034, P < 0.001). LAG-3 was expressed in murine tumor-infiltrating lymphocytes, and its blockade decreased tumor growth and enhanced secretion of interferon-gamma by CD8^+ and CD4^+ T cells.Conclusions: LAG-3 blockade may be a promising strategy to improve the effects of targeted therapy in STS.

类器官芯片在纳米药物递送系统研究中的应用及前景

类器官是源自人类干细胞、器官特异性祖细胞及分离肿瘤组织的自组装三维多细胞微型器官模型,可在体外重现器官的关键结构特征与生理机能。微流控芯片作为类器官的体外培养平台,为研究细胞在异质群体和微环境中的行为提供机会,因此,成为助力人类器官发育研究、疾病建模及药物筛选的强大工具。基于先进纳米技术构建的药物载体可以显著提升疾病治疗效果,是目前新药物研发和临床治疗的焦点。利用类器官芯片对纳米递送系统的安全性和有效性进行评估,不仅有助于了解药物递送载体与靶点微环境的相互作用机制,科学指导纳米载体的设计;也在患者个性化精准医疗领域具有应用价值。本文讨论了传统二维细胞模型以及动物模型评价纳米药物递送系统的局限性,分析了用类器官芯片作为体外评价平台的优势。在此基础上,总结了近年来类器官芯片在纳米药物递送领域的应用现状与发展方向,并对其前景进行了展望。

Tertiary lymphoid structures in cancer – considerations for patient prognosis

Tertiary lymphoid structures(TLS)are ectopic lymphoid formations that form within nonlymphoid tissue.They share structural and functional characteristics with secondary lymphoid structures such as lymph nodes and can contain B-cell follicles and germinal centers surrounded by a T-cell region.TLS have been described in several types of cancers and are usually associated with positive patient outcomes.However,TLS differ vastly in cellular composition and location within tissue types.In this review,we discuss factors confounding the interpretation of the evidence for a prognostic role for TLS in cancer and frame these factors in the context of translation to regular clinical use.

急性脑梗死患者阿替普酶静脉溶栓后神经功能和预后与血生化常见指标的相关性分析

目的探讨急性脑梗死患者阿替普酶静脉溶栓后神经功能、预后与血清尿酸、同型半胱氨酸(Hcy)及低密度脂蛋白胆固醇(LDL-C)的相关性。方法选择2020年1月至2022年12月于长沙市第一医院神经重症监护室收治的行溶栓治疗的急性脑梗死患者220例,根据溶栓后3个月改良的Rankin量表(mRS)评分分为预后不良组91例(mRS评分>2分)和预后良好组129例(mRS评分≤2分)。比较2组血清尿酸、Hcy及LDL-C水平,分析其与患者溶栓后美国国立卫生研究院卒中量表(NIHSS)评分相关性、对溶栓后预后不良的预测价值。结果与溶栓前比较,2组溶栓后1、3 d血清尿酸、Hcy、LDL-C水平及NIHSS评分明显降低,且溶栓后3 d血清尿酸、Hcy水平及NIHSS评分明显低于溶栓后1 d,差异有统计学意义(P<0.05)。预后不良组溶栓后1、3 d血清尿酸、Hcy、LDL-C水平及NIHSS评分明显高于预后良好组,差异有统计学意义(P<0.05,P<0.01)。Person相关性分析显示,溶栓后1、3 d血清尿酸、Hcy、LDL-C水平与NIHSS评分呈正相关(P<0.01)。ROC曲线分析显示,溶栓后1 d血清尿酸、Hcy、LDL-C曲线下面积分别为0.707(95%CI:0.639~0.776)、0.800(95%CI:0.739~0.860)、0.624(95%CI:0.550~0.698),溶栓后3 d血清尿酸、Hcy、LDL-C曲线下面积分别为0.655(95%CI:0.583~0.726)、0.730(95%CI:0.664~0.795)、0.573(95%CI:0.497~0.649)。结论急性脑梗死溶栓后预后不良患者血清尿酸、Hcy及LDL-C水平呈升高趋势,还与神经损伤程度存在关系,溶栓后1 d指标水平对溶栓后不良预后有较好的预测价值。

UBE2S在食管鳞状细胞癌组织中的表达及临床意义

目的:探讨食管鳞状细胞癌(ESCC)组织中UBE2S的表达及临床意义。方法:采用免疫组化SP法检测ESCC、上皮内瘤变中UBE2S的表达情况,分析其表达与ESCC患者临床病理特征的相关性及对预后的影响。结果:UBE2S在ESCC中的表达明显高于癌旁组织(P=0.035)。高级别上皮内瘤变(HIN)(P<0.001)及ESCC(P<0.001)的阳性率均高于瘤旁组织。HIN(P=0.016)及ESCC(P=0.008)的阳性率均高于低级别上皮内瘤变(LIN)。ESCC患者中UBE2S表达与种族有关(P<0.001),UBE2S表达与TP53(r=0.288,P<0.001)、Ki67(r=0.210,P=0.002)表达呈正相关。UBE2S表达是影响ESCC患者无进展生存期(P=0.005)和总生存期(P=0.005)的独立因素。结论:UBE2S高表达可能是ESCC患者不良预后的独立因素,而GEPIA上的UBE2S与ESCC预后无统计学意义,有待进一步研究。

Cytotoxic CD8+ T cells and tissue resident memory cells in colorectal cancer based on microsatellite instability and BRAF status

BACKGROUND Recent studies in non-colorectal malignancy have associated T resident memory(T_(RM)) cells with improved patient survival. It is unknown if T_(RM) plays a role in colorectal cancer(CRC).AIM To examine the potential role of T_(RM) cells in providing immunogenicity in CRC stratified by microsatellite instability(MSI) and BRAF status.METHODS Patients with known MSI and BRAF mutation status were eligible for inclusion in this study. CRC tumour sections stained with haematoxylin and eosin were microscopically reviewed and the images scanned prior to assessment for location of invading edge and core of tumour. Sequential sections were prepared for quantitative multiplex immunohistochemistry(IHC) staining. Opal Multiplex IHC staining was performed with appropriate positive and negative controls and imaged using a standard fluorescent microscope fitted with a spectral scanning camera(Mantra) in conjunction with Mantra snap software. Images were unmixed and annotated in in Form 2.2.0. Statistical analysis was performed using Graphpad Prism Version 7 and Stata Version 15.RESULTS Seventy-two patients with known MSI and BRAF status were included in the study. All patients were assessed for MSI by IHC and high resolution capillary electrophoresis testing and 44 of these patients successfully underwent quantitative multiplex IHC staining. Overall, there was a statistically significant increase in CD8+ T_(RM) cells in the MSI(BRAF mutant and wild type) group over the microsatellite stable(MSS) group. There was a statistically significant difference in CD8+ T_(RM) between high level MSI(MSI-H):BRAF mutant [22.57, 95% confidence interval(CI): 14.31-30.84] vs MSS [8.031(95%CI: 4.698-11.36)], P = 0.0076 and MSI-H:BRAF wild type [16.18(95%CI: 10.44-21.93)] vs MSS [8.031(95%CI: 4.698-11.36)], P = 0.0279. There was no statistically significant difference in CD8 T cells(both CD8+CD103-and CD8+CD103+T_(RM)) between MSI-H: BRAF mutant and wild type CRC.CONCLUSION This study has shown that CD8+ T_(RM) are found in greater abundance in MSI-H CRC, both BRAF mutant and MSI-H:BRAF wild type, when compared with their MSS counterpart. CD8+ T_(RM) may play a role in the immunogenicity in MSI-H CRC(BRAF mutant and BRAF wild type). Further studies should focus on the potential immunogenic qualities of T_(RM) cells and investigate potential immunotherapeutic approaches to improve treatment and survival associated with CRC.

静脉溶栓联合Solitaire FR型支架取栓术对急性大动脉闭塞性脑梗死患者的疗效分析

目的探究静脉溶栓联合Solitaire FR型支架取栓术对急性大动脉闭塞性脑梗死患者血管再通、神经功能的影响及预后分析。方法回顾性选取2020年10月至2023年3月南京市溧水区人民医院收治的急性大动脉闭塞性脑梗死患者172例,根据治疗方法分为对照组和研究组,每组86例,对照组采用阿替普酶静脉溶栓治疗,研究组采用阿替普酶静脉溶栓联合Solitaire FR型支架取栓术治疗。比较2组血管再通、神经功能、脑血流灌注和不良事件情况,随访90 d采用改良Rankin量表评价预后。结果2组成功再通率比较,差异无统计学意义(P>0.05),研究组完全再通率高于对照组(68.60%vs 50.00%,P<0.05)。治疗后7 d、14 d,研究组美国国立卫生研究院卒中量表评分低于对照组,差异有统计学意义(P<0.01)。治疗后,研究组脑血容量、脑血流量高于对照组,平均通过时间低于对照组,差异有统计学意义(P<0.05,P<0.01);2组治疗期间不良事件总发生率比较,差异无统计学意义(P>0.05)。随访90 d,研究组良好预后比例高于对照组(80.23%vs 63.95%,P<0.05)。结论静脉溶栓联合Solitaire FR型支架取栓术在急性大动脉闭塞性脑梗死中效果较好,血管再通效果理想,可明显改善患者神经功能和预后。