Strategies that enhance the function of chimeric antigen receptor-modified T (CAR-T) cells for solid tumors are critical. Inhibitory immuno-checkpoints blockade could potentially enhance CAR-T cell function. TIM-3 is an important negative regulator of T cell activity, but whether TIM-3 blockade could affect CAR-T cell function remains unclear. In our study, we successfully constructed TIM-3-silenced CAR-T cells by dual-promoter lentivirus vectors that simultaneously express the TIM-3 targeting short hairpin RNA (shRNA) and a third-generation CAR recognizing HER2. We demonstrated that down-regulation of TIM-3 did not affect the phenotype of CAR-T cells. CAR-T cells with TIM-3 blockade exhibited higher lytic cytotoxicity to target cells in vitro. Additionally, TIM-3-silenced CAR-T cells displayed robust anti-tumor activity in a murine xenograft model, which is comparable to standard CAR-T cells. Our study demonstrates the effect of down-regulation of immune checkpoint TIM-3 on the anti-tumor function of CAR T cells, providing new ideas for improving the potency of CAR-T cell therapies in solid tumors.

Background and Aims: The differential diagnosis of benign and malignant deep lymph nodes (LNs) has been a significant challenge up until now. Endoscopic ultrasound (EUS) elastography is a real-time imaging technique evaluated in several studies with diverse results. A meta-analysis was performed to assess the performance of EUS elastography for the differentiation of benign and malignant deep LNs. Methods: All the eligible studies were searched by PubMed, Medline, Embase, and the Cochrane Library. A meta-analysis was performed to obtain pooled sensitivity, specificity, positive and negative likelihood ratio (LR). The diagnostic odds ratio (DOR) and area under the curve (AUC) were used to examine the accuracy of qualitative and quantitative EUS elastography. Results: A total of eleven studies including 599 patients and 943 LNs were analyzed. In studies using the qualitative color pattern as the diagnostic standard, the pooled sensitivity, specificity, positive and negative LR were 0.82 [95% confidence interval (CI): 0.77-0.86)], 0.92 (95%CI: 0.89-0.94), 7.83 (95%CI: 3.58-17.09) and 0.22 (95%CI: 0.14-0.34), respectively. In studies using the quantitative value as the diagnostic standard, the pooled sensitivity, specificity, positive and negative LR were 0.80 (95%CI: 0.74-0.86) and 0.87 (95%CI: 0.82-0.92), 5.63 (95%CI: 3.46-9.17) and 0.23 (95%CI: 0.12-0.44), respectively. The summary DOR and the AUC were 48.73 (95%CI: 21.83-108.80) and 0.933 (Q*=0.869) for qualitative EUS elastography, 33.35 (95% CI: 12.81-86.87) and 0.923 (Q*=0.857) for quantitative EUS elastography. Conclusions: Our meta-analysis shows that both qualitative and quantitative EUS elastography have high accuracy in the detection of malignant deep LNs, which could be used as a valuable complementary method to EUS-FNA for the differentiation of deep LNs in the future.

The current study aimed to investigate whether previous abdominal surgery (PAS) could affect the outcomes of colorectal cancer (CRC) surgery. We conducted the search strategy in three databases (PubMed, Embase, and the Cochrane Library) from inception to May 26, 2022. The short-term and long-term outcomes were compared between the PAS group and the non-PAS group. Odds ratios (ORs) and 95

Objective To investigate the killing efficacy of chimeric antigen receptor T (CAR-T) cells targeting epithelial cellular adhesion molecule (EpCAM) in several hepatoma cell lines. Methods Flow cytometry was used to detect the expression of EpCAM in hepatoma cells (SK-hep1, Hep-G2, HuH7 and BEL-7402). After EpCAM -targeting CAR-T cells were constructed using lentiviral vectors with CAR genes, flow cytometry was used to detect the expression of EpCAM-CAR in the obtained cells, and to analyze the CD3/4/8 phenotype of CAR-T cells without transfection of lentiviral vectors. Then the effector cells and target cells were co-cultured, and LDH release assay and ELISA were used to detect the release of LDH and major cytokines in the supernatant. Results EpCAM was highly expressed in Hep-G2, HuH7 and BEL-7402 cells, but not in SK-hep1 cells. Flow cytometry showed that the expression rate of EpCAM-CAR was 43.21% in CAR-T cells after lentivirus infection. Flow cytometry also indicated that up to 95.1% of peripheral blood mononuclear cells (PBMCs) were CD3 positive, of which 61.40% were CD8+ T cells and 32.13% were CD4+ T cells. LDH release assay revealed that the obtained CAR-T cells had stronger direct killing ability on EpCAM-positive hepatoma cell lines Hep-G2, HuH7, and BEL-7402. ELISA showed that EpCAM-positive target cells Hep-G2, HuH7, and BEL-7402 induced CAR-T cells to release more IFN-γ and TNF-α. For SK-hep1 cells with negative EpCAM expression, IFN-γ and TNF-α releases were not significantly different between the CAR-T group and the control group, and the levels of these 2 cytokines werw increased with the increase of E ∶T ratio. Results Compared with ordinary CAR-T cells, EpCAM-targeting CAR-T cells have a stronger killing effect on EpCAM-positive hepatocellular carcinoma cells.

Crizotinib is an oral [anaplastic lymphoma kinase](https://en.wikipedia.org/wiki/Anaplastic_lymphoma_kinase) and [c-ros oncogene 1](https://en.wikipedia.org/wiki/C-ros_oncogene_1) inhibitor, which is approved by the Food and Drug Administration for the treatment of metastatic non-small cell lung cancer (NSCLC). The 2011 Clinical trials demonstrated excellent treatment response, survival, and few side effects. We report a case of severe ulcerative oesophagitis with stenosis that was not reported in the 2011 Clinical trials and earlier studies.

上消化道早癌筛查技能教学是目前内镜教学培训的空缺点和难点.本研究以来自基层已掌握基本胃镜操作技术的进修内镜医师为教学对象,建立了关于上消化道早癌诊断的规范化培训方案.培训周期共12周;教学内容包括理论培训、早癌意识培训、规范的胃镜操作技术、早期食管癌的内镜诊断、早期胃癌的内镜诊断,以及理论和技能结业考核;考核通过后进行早癌病例比赛.通过问卷调查,100％(15/15)的学员认为上消化道早癌诊断规范化培训用于提高上消化道早癌检出率是必要的,且能明显提高学员上消化道早癌的诊断水平.因此,本研究上消化道早癌诊断规范化培训流程总体设置合理,值得进一步推广.

Endoscopic ultrasonography (EUS) is useful for the diagnosis of gastrointestinal stromal tumors (GISTs), but is limited by subjective interpretation. Studies on artificial intelligence (AI)-assisted diagnosis are under development. Here, we used a meta-analysis to evaluate the diagnostic performance of AI in the diagnosis of GISTs using EUS images. PubMed, Ovid Medline, Embase, Web of science, and the Cochrane Library databases were searched for studies based on the EUS using AI for the diagnosis of GISTs, and a meta-analysis was performed to examine the accuracy. Overall, 7 studies were included in our meta-analysis. A total of 2431 patients containing more than 36,186 images were used as the overall dataset, of which 480 patients were used for the final testing. The pooled sensitivity, specificity, positive, and negative likelihood ratio (LR) of AI-assisted EUS for differentiating GISTs from other submucosal tumors (SMTs) were 0.92 (95

A 59-year-old man was admitted with a 1-year history of recurrent diarrhea and abdominal pain. Colonoscopy performed the previous year showed multiple colonic ulcers, which were confirmed by pathology. Repeat colonoscopy demonstrated a sigmoid ulcer with stenosis that could not be traversed with an adult colonoscope.

该文主要研究慢病毒介导的T细胞免疫球蛋白黏液素3(T-cell immunoglobulin mucin-3,Tim-3)对宫颈癌细胞的促肿瘤作用及其相关作用机制.首先,采用PCR方法获取Tim-3片段,将其克隆入pCDH载体中,并通过酶切和测序进行鉴定;将三个表达载体,即重组阳性表达载体(pCDH-Tim-3)、空载体(pCDH-NC)和绿色荧光蛋白载体(pCDH-GFP),分别与psPAX2和pMD2.G共转染至293T细胞中,从而形成相应的慢病毒组.然后,将三组慢病毒分别感染HeLa细胞,获得稳定细胞株(HeLa-Tim-3、HeLa-NC、HeLa-GFP),其中设置HeLa-NC为空载病毒阴性对照组;最后,通过观察HeLa-GFP组,初步获取慢病毒感染效率;运用流式细胞术、Western blot和细胞免疫荧光检测Tim-3的相对表达水平;细胞划痕和Transwell检测细胞的迁移及侵袭能力;Westem blot检测上皮间质转化(epithelial-to-mesenchymal,EMT)相关蛋白(E-cadherin、N-cadherin、Snail)的表达变化;流式细胞术检测细胞凋亡率变化.该文成功构建了pCDH-Tim-3慢病毒表达载体并将其包装成慢病毒,获得对应的稳定细胞株;在HeLa-GFP中可以观察到大量的绿色荧光;与HeLa-NC组相比,Western blot、流式细胞术和细胞免疫荧光检测结果均表明,HeLa-Tim-3组的Tim-3蛋白表达水平显著升高(P＜0.05);细胞划痕实验和Transwell检测结果均表明,HeLa-Tim-3组的迁移侵袭能力明显提高(P＜0.01);另外,Western blot检测结果表明,与HeLa-NC组相比,HeLa-Tim-3组N-cadherin和Snail显著增多但E-cadherin明显减少(P＜0.05),而流式细胞术检测结果表明,HeLa-Tim-3组凋亡率显著减少(P＜0.001).因此,高表达Tim-3宫颈癌细胞可以通过EMT转化和抑制细胞凋亡等相关作用机制来加强肿瘤的恶性进展.

Chimeric antigen receptor-modified T cells (CAR-T cells) have emerged as a promising cancer immunotherapy for solid tumors. Epithelial cell adhesion molecule (EpCAM) is overexpressed in a variety of tumors and is recognized as a biomarker for circulating tumor cells and cancer stem cells, representing an attractive target for adoptive T-cell immunotherapy. This study generated third-generation CAR-T cells with redirected specificity to EpCAM (EpCAM CAR-T) by lentiviral vector. The study demonstrated that EpCAM CAR-T cells can elicit lytic cytotoxicity to target cells in an EpCAM-dependent manner and secrete cytotoxic cytokines, including interferon gamma and tumor necrosis factor alpha. Furthermore, adoptive transfer of EpCAM CAR-T cells significantly delayed tumor growth and formation in xenograft models. In addition, the safety evaluation showed that CAR-T cells have no systemic toxicity in mice. The data confirmed the antitumor ability and safety of CAR-T cells targeting EpCAM and may provide a new target for CAR-T cell therapies in treating solid tumors.

Background: Studies have shown mixed results on the role of postoperative adjuvant radiotherapy (PORT) in surgically managed locally advanced laryngeal cancer. Objectives: The aim of this study is to review and investigate the role of PORT in patients with locally advanced laryngeal cancer using meta-analysis. Materials and methods: Relevant studies were searched using PubMed and eligible information has been extracted. Then, meta-analysis of hazard ratio (HR) was performed to evaluate the role of PORT in locally advanced laryngeal cancer. Results: This meta-analysis included 7 published studies containing 2007 patients. For overall survival (OS), patients of locally advanced laryngeal cancer who were treated with PORT have a combined hazard ratio (HR) of 0.67 with 95%CI (0.56, 0.79), compared to those who were not treated with PORT, which was significantly associated with better survival. PORT was also associated with a better disease-free survival (DFS) and local control rate (LCR) in patients with locally advanced laryngeal cancer. The pooled HR and 95%CI for DFS and LCR were 0.72 (0.53, 0.99) and 0.29 (0.09, 0.99), respectively.

Background The optimal treatment in the third-line and later-line setting for metastatic colorectal cancer (mCRC) has not been established. As reported, regorafenib and fruquintinib have shown to be superior to placebo in mCRC. However, no direct clinical comparison of regorafenib and fruquintinib has been conducted; we performed a systematic review and network meta-analysis to compare the efficacy and safety of regorafenib and fruquintinib. Methods PubMed, Embase, and the Cochrane Library were systematically searched and randomized-controlled trials (RCTs) assessing the effect and safety of regorafenib or fruquintinib versus placebo for patients with mCRC were included. Two investigators independently searched articles, extracted data, and assessed the quality of included studies. After that, we performed pairwise direct meta-analyses (regorafenib vs. placebo and fruquintinib vs. placebo) and indirect comparison (regorafenib vs. fruquintinib) using network meta-analyses methods. Results Three RCTs involving 1380 patients were included in the meta-analysis. In the direct meta-analysis, regorafenib and fruquintinib both showed survival benefits when compared with placebo. For the indirect comparison, fruquintinib shows no significant difference in OS compared to regorafenib (HR 0.97; 95% CI 0.64-1.46). Regarding PFS, there was a tendency that fruquintinib was superior to regorafenib (HR 0.65; 95% CI 0.39-1.08); however, there was no statistic difference. For the safety analysis, in indirect comparison, fruquintinib showed significant difference in all-grade toxicity compared to regorafenib (OR 0.73; 95% CI 0.65-0.82), especially in subgroup of proteinuria (OR 0.31; 95% CI 0.11-0.86). For the grade 3-5 toxicity, fruquintinib showed no significant difference when compared with regorafenib (OR 0.92; 95% CI 0.64-1.32). Conclusion Based on efficacy and safety, there was a tendency that fruquintinib was superior to regorafenib, as a whole, regorafenib and fruquintinib demonstrated similar clinical benefit for patients with refractory mCRC. It seems that fruquintinib has less toxic in all-grade toxicity when compared with regorafenib.

Background and aims: Endoscopic ultrasound (EUS) elastography is a novel non-invasive technique that can be used for distinguishing benign from malignant pancreatic masses. However, the studies have reported widely varied sensitivities and specificities. A meta-analysis was performed to assess the performance of EUS elastography for the differentiation of benign and malignant pancreatic masses. Methods: All the eligible studies were searched by PubMed, Medline, Embase, and the Cochrane Library. Sensitivity, specificity, positive likelihood ratio (LR), negative LR, and area under the curve (AUC) were calculated to examine the accuracy. Results: A total of nineteen studies which included 1687 patients were analyzed. The pooled sensitivity and specificity for the diagnosis of malignant pancreatic masses were 0.98 (95% confidence interval [CI] 0.96-0.99) and 0.63 (95% CI 0.58-0.69) for qualitative EUS elastography, 0.95 (95% CI 0.93-0.97) and 0.61 (95% CI 0.56-0.66) for quantitative EUS elastography, respectively. The positive and negative LR were 2.60 (95% CI 1.84-3.66) and 0.05 (95% CI 0.02-0.10) for qualitative EUS elastography, 2.64 (95% CI 1.82-3.82) and 0.10 (95% CI 0.06-0.16) for quantitative EUS elastography, respectively. The summary diagnostic odds ratio (DOR) and the AUC were 60.59 (95% CI 28.12-130.56) and 0.91 (Q* = 0.842) for qualitative EUS elastography, 30.09 (95% CI 15.40-58.76) and 0.93 (Q* = 0.860) for quantitative EUS elastography. Conclusions: Our meta-analysis shows that both qualitative and quantitative EUS elastography have high accuracy in the detection of malignant pancreatic masses, which could be used as a valuable complementary method to EUS-FNA for the differentiation of pancreatic masses in the future. (C) 2018 IAP and EPC. Published by Elsevier B.V. All rights reserved.

The high incidence of metastasis accounts for most of the lethality of ovarian cancer. Invadopodia are small, specialized types of machinery that degrade the extracellular matrix and are thus involved in the invasion and metastasis of cancer cells. The formation of invadopodia is regulated by both genetic and epigenetic factors. However, the ways by which methylation/demethylation regulates the dynamics of invadopodia in ovarian cancer are largely unknown. In this study, we found that the inhibition of methylation by 5-AZ (5-Azacytidine) increased the formation of invadopodia and enhanced degradation of the extracellular matrix in ovarian cancer cells. In mouse xenograft models, treatment with 5-AZ increased the number of metastatic nodules, which suggests an elevated potential for metastasis by demethylation. Further investigation indicated that the inhibition of methylation elevated the transcription of PIK3CA and upregulated genes involved in the PI3K-AKT signaling pathway. In addition, this induction likely occurs though the epigenetic regulation of PIK3CA because analyses of the DNA methylation level of the PIK3CA promoter region found that 5-AZ treatment decreased the methylation of CpG islands in SKOV3 and A2780 cells. Our study demonstrated that epigenetic factors regulate the metastatic potential of ovarian cancer cells and provide rationale for therapies that inhibit PI3K- invadopodia-mediated metastasis.

Recent reports on the impressive efficacy of chimeric antigen receptor (CAR)-modified T cells against hematologic malignancies have inspired oncologists to extend these efforts for the treatment of solid tumors. Clinical trials of CAR-T-based cancer immunotherapy for solid tumors showed that the efficacies are not as remarkable as in the case of hematologic malignancies. There are several challenges that researchers must face when treating solid cancers with CAR-T cells, these include choosing an ideal target, promoting efficient trafficking and infiltration, overcoming the immunosuppressive microenvironment, and avoiding associated toxicity. In this review, we discuss the obstacles imposed by solid tumors on CAR-T cell-based immunotherapy and strategies adopted to improve the therapeutic potential of this approach. Continued investigations are necessary to improve therapeutic outcomes and decrease the adverse effects of CAR-T cell therapy in patients with solid malignancies in the future.

T cells, genetically modified by chimeric antigen receptors (CAR-T), are endowed with specificity to a desired antigen and are cytotoxic to cells expressing the targeted antigen. CAR-T-based cancer immunotherapy is a promising therapy for curing hematological malignancy, such as acute lymphoid leukemia, and is promising for extending their efficacy to defeat solid tumors. To date, dozens of different CAR-T cells have been evaluated in clinical trials to treat tumors; this necessitates the establishment of guidelines for the production and application of CAR-T cells. However, it is challenging to standardize CAR-T cancer therapy because it involves a combination of gene therapy and cell therapy. In this review, we compare the existing guidelines for CAR-T cells and discuss the challenges and considerations for establishing guidance for CAR-T-based cancer immunotherapy.

BackgroundNeoadjuvant therapy is administered to breast cancer patients as an induction process before surgery or radiotherapy to reduce tumor size. Human epidermal growth factor receptor-2 (HER-2) negative breast cancer lacks effective standard target therapy. Bevacizumab has a controversial role in the treatment of breast cancer and we conduct a meta-analysis to evaluate the value of adding bevacizumab in neoadjuvant regimen.MethodsPotentially eligible studies were retrieved using PubMed, EMBASE and Medline. Clinical characteristics of patients and statistical data with pathological complete response (pCR) data were collected. Then a meta-analysis model was established to investigate the correlation between administration of bevacizumab in neoadjuvant therapy and pCR rates in HER2 negative breast cancer.ResultsSeven eligible studies and 5408 patients were yielded. The pCR rates for "breast" or "breast plus lymph node" were similar. In subgroup analysis, we emphasized on patients with triple-negative breast cancer (TNBC). In the criterion of "lesions in breast" the pooled ORs was 1.55 [1.29, 1.86], P<0.00001 and regarding to the evaluation criterion of "lesions in breast and lymph nodes", the pooled ORs was 1.48 [1.23, 1.78], P<0.0001, in favor of bevacizumab administration.ConclusionAccording to our pooled results, we finally find that bevacizumab addition as a neoadjuvant chemotherapy component, for induction use with limited cycle to improve the pCR rates and patients may avoid long-term adverse event and long-term invalid survival improvement. Especially in subgroup analysis, pCR rates could be improved significantly and physicians could consider bevacizumab with caution. As patients could avoid the adverse event caused by long-term using of bevacizumab, long-term quality of life improvement may be achieved, especially in TNBC.

ABSTRACT Purpose The use of contrast‐enhanced sonography (CEUS) has yielded promising results in the differentiation of thyroid nodules. We conducted this meta‐analysis to assess its performance in identifying and distinguishing between benign and malignant thyroid nodules. Methods PubMed, Medline, Embase, and the Cochrane Library were searched for studies published through the end of December 2013. Sensitivity, specificity, positive and negative likelihood ratios, diagnostic odds ratio, and area under the curve were calculated. Results A total of 13 studies were included in this meta‐analysis. For the diagnosis of malignant thyroid nodules worldwide, the overall mean rates of sensitivity and specificity of CEUS were 90% (95% confidence interval [CI], 88–93%) and 86% (95% CI, 83–89%), respectively. The summary diagnostic odds ratio was 52.83 (95% CI, 21.71–128.55), and the area under the curve for the summary receiver operating characteristic curve was 0.94 (95% CI, 0.90–0.98). Conclusions This meta‐analysis indicates that CEUS may be a valuable supplemental method, with high rates of sensitivity and specificity, to use for identifying and distinguishing between benign and malignant thyroid nodules. © 2015 Wiley Periodicals, Inc. J Clin Ultrasound 44 :199–209, 2016

OBJECTIVE:To compare the efficacy of radiotherapy (RT) plus chemotherapy (CMT) versus RT alone for early stage nasal natural killer (NK)/T-cell lymphoma.METHODS:All the eligible studies were searched by PubMed, Medline, Embase and the Cochrane Library. The meta-analysis was performed to compare odds ratios (ORs) for overall survival (OS), disease-free survival (DFS) and progression-free survival (PFS).RESULTS:Eight studies were included in the meta-analysis. Chemotherapy group did not significantly differ from RT group. The pooled OR and 95% confidence interval (CI) for 1-year, 3-year, 5-year and 10-year OS was 1.25 [0.84, 1.87], 1.10 [0.76, 1.58], 0.83 [0.59, 1.17] and 1.05 [0.70, 1.56]. In addition, the combined OR and 95% CI for 5-year DFS and PFS were 0.96 [0.53, 1.73] and 0.71 [0.45, 1.12].CONCLUSIONS:The current evidence suggests that CMT was not superior to RT alone. Radiotherapy may be still the main method in the treatment of early stage nasal NK/T-cell lymphoma.

Cancer-associated fibroblasts (CAFs), key components of the tumor stroma, can regulate tumorigenesis by altering the tumor microenvironment in variety of ways to promote angiogenesis, recruit inflammatory immune cells and remodel the extracellular matrix. Using a murine xenograft model of colon carcinoma, the present study observed that oxaliplatin increased the accumulation of CAFs and stimulated the production of cytokines associated with CAFs. When oxaliplatin was combined with the small-molecule dipeptidyl peptidase inhibitor PT-100, which inhibits CAFs by targeting fibroblast activation protein (FAP), the accumulation of CAFs was markedly reduced, xenograft tumor growth was significantly suppressed and the survival of the mice increased, compared to those of mice treated with oxaliplatin or PT-100 alone. Furthermore, the xenograft tumor tissues of mice treated with oxaliplatin and PT-100 contained lower numbers of tumor-associated macrophages and dendritic cells, expressed lower levels of cytokines associated with CAFs and had a lower density of CD31+ endothelial cells. The present study demonstrated that pharmacological inhibition of CAFs improved the response to chemotherapy, reduced the recruitment of immune tumor-promoting cells and inhibited angiogenesis. Combining chemotherapy with agents which target CAFs may represent a novel strategy for improving the efficacy of chemotherapy and reducing chemoresistance.