Tumor-associated macrophages (TAMs) play a crucial role in colorectal cancer (CRC) metastasis, particularly in the establishment of the liver metastatic niche. TAMs facilitate metastasis by enabling epithelial-mesenchymal transition (EMT) and suppressing anti-tumor immunity. While their roles in primary tumors have been extensively investigated, their functions at metastatic sites remain poorly understood. In this study, we analyzed the single-cell RNA sequencing data from 46 CRC patients, encompassing 402,972 cells, to identify macrophage subsets associated with liver metastasis. A distinct population of APOC1+ IFI30+ TAMs was identified and found to be significantly enriched in metastatic liver lesions. Compared to primary tumors (PT), the number of APOC1+ IFI30+ TAMs significantly increased at liver metastatic sites after CRC cells metastasized to the liver. We employed spatial transcriptomics to validate the above conclusions. These TAMs exhibited significantly enhanced immunosuppressive capacity and cholesterol synthesis metabolism. Further investigation showed that TAMs within metastatic niches reprogram the mevalonate pathway to enhance cholesterol synthesis, characterized by upregulation of key genes such as HMGCR and ACAT1/2. These TAMs act on surrounding tumor cells by secreting cholesterol and cytokines such as TNFSF12, TGFB1, MIF and LGALS9, thereby enhancing tumor adhesion and proliferation and creating an immunosuppressive microenvironment to continuously support tumor growth. Our findings provided new insights into the complex interactions among TAMs, immune cells, and tumor cells within CRC metastatic microenvironment, highlighting potential therapeutic targets to disrupt TAMs-mediated immune evasion and metastatic progression.

The underlying molecular mechanism of patients with colorectal liver metastasis (CRLM) remains unclear. In this study, it is found that cancer-associated fibroblasts (CAFs)-derived extracellular vesicles (EVs) are significantly enriched in circTAX1BP1 in CRLM, which associate with poor prognosis. The disruption of EV-packaged circTAX1BP1 significantly inhibits CRLM in vivo and in vitro. Mechanistically, CAF-derived EV-packaged circTAX1BP1 is delivered to colorectal cancer (CRC) cells, where it binds to VIRMA and promotes its lactylation at lysine residue 1713 by recruiting AARS2. Lactylated VIRMA enhances m6A-based modification and stability of SP1 mRNA. SP1 mediates the transcription of TGF-β, enhancing epithelial-mesenchymal transition and paracrine TGF-β of CRC cells. Notably, this study identifies an important subgroup ITGA11+ myCAFs through single-cell RNA sequencing data. Paracrine TGF-β of CRC cells specifically targets ITGA11+ myCAFs, activating the TGF-β signalling pathway, which contributes to extracellular matrix remodeling and increases delivery of EV-packaged circTAX1BP1, forming a positive feedback loop to promote CRLM. Finally, the combined blockade of EV-packaged circTAX1BP1 and TGF-β can effectively disrupt this feedback loop and significantly inhibit tumor progression in a PDX model. Overall, this study provides an in-depth understanding of tumor cell-CAFs crosstalk and new insights into therapeutic targets for CRLM.

BackgroundColorectal cancer is the third most prevalent cancer across the globe. Despite a diversity of treatment methods, the recurrence and mortality rates of the disease remain high. Recent studies have revealed a close association of the gut microbiota with the occurrence, development, treatment response, and prognosis of colorectal cancer.ObjectiveThis study aims to integrate transcriptome and microbiome data to identify colorectal cancer subtypes associated with different gut microbiota and evaluate their roles in patient survival prognosis, tumor microenvironment (TME), and drug treatment response.MethodsAn integrated analysis of microbiome data was conducted on samples of colorectal cancer from public databases. Based on this, two tumor subtypes (C1 and C2) closely associated with patient survival prognosis were identified and a risk score model was constructed. The survival status, clinical parameters, immune scores, and other features were analyzed in-depth, and the sensitivity of various potential drugs was examined.ResultsA thorough examination of microbiome information obtained from colorectal cancer patients led to the identification of two primary tumor clusters (C1 and C2), exhibiting notable variations in survival outcomes. Patients with the C1 subtype were closely associated with better prognosis, while those with the C2 subtype had higher gut microbial richness and poorer survival prognosis. A predictive model utilizing the microbiome data was developed to accurately forecast the survival outcome of patients with colorectal cancer. The TME scores provided a biological basis for risk assessment in high-risk (similar to the C2 subtype) patient cohorts. Evaluation of the sensitivity of different subtypes to various potential drugs, indicated the critical importance of personalized treatment. Further analysis showed good potential of the developed risk-scoring model in predicting immune checkpoint functions and treatment response of patients, which may be crucial in guiding the selection of immunotherapy strategies for patients with colorectal cancer.ConclusionThis study, through a comprehensive analysis of colorectal cancer microbiome, immune microenvironment, and drug sensitivity, enhances the current understanding of the multidimensional interactions of colorectal cancer and provides important clinical indications for improving future treatment strategies. The findings offer a new perspective on improving treatment response and long-term prognosis of patients with CRC through the regulation of microbiota or the utilization of biomarkers provided by it.

Tumor-associated macrophages (TAM) subtypes have been shown to impact cancer prognosis and resistance to immunotherapy. However, there is still a lack of systematic investigation into their molecular characteristics and clinical relevance in different cancer types. Single-cell RNA sequencing data from three different tumor types were used to cluster and type macrophages. Functional analysis and communication of TAM subpopulations were performed by Gene Ontology-Biological Process and CellChat respectively. Differential expression of characteristic genes in subpopulations was calculated using zscore as well as edgeR and Wilcoxon rank sum tests, and subsequently gene enrichment analysis of characteristic genes and anti-PD-1 resistance was performed by the REACTOME database. We revealed the heterogeneity of TAM, and identified eleven subtypes and their impact on prognosis. These subtypes expressed different molecular functions respectively, such as being involved in T cell activation, apoptosis and differentiation, or regulating viral bioprocesses or responses to viruses. The SPP1 pathway was identified as a critical mediator of communication between TAM subpopulations, as well as between TAM and epithelial cells. Macrophages with high expression of SPP1 resulted in poorer survival. By in vitro study, we showed SPP1 mediated the interactions between TAM clusters and between TAM and tumor cells. SPP1 promoted the tumor-promoting ability of TAM, and increased PDL1 expression and stemness of tumor cells. Inhibition of SPP1 attenuated N-cadherin and β-catenin expression and the activation of AKT and STAT3 pathway in tumor cells. Additionally, we found that several subpopulations could decrease the sensitivity of anti-PD-1 therapy in melanoma. SPP1 signal was a critical pathway of communication between macrophage subtypes. Some specific macrophage subtypes were associated with immunotherapy resistance and prognosis in some cancer types.

Cancer cachexia-associated muscle wasting as a multifactorial wasting syndrome, is an important factor affecting the long-term survival rate of tumor patients. Photobiomodulation therapy (PBMT) has emerged as a promising tool to cure and prevent many diseases. However, the effect of PBMT on skeletal muscle atrophy during cancer progression has not been fully demonstrated yet. Here, we found PBMT alleviated the atrophy of myotube diameter induced by cancer cells in vitro, and prevented cancer-associated muscle atrophy in mice bearing tumor. Mechanistically, the alleviation of muscle wasting by PBMT was found to be involved in inhibiting E3 ubiquitin ligases MAFbx and MuRF-1. In addition, transcriptomic analysis using RNA-seq and GSEA revealed that PI3K/AKT pathway might be involved in PBMT-prevented muscle cachexia. Next, we showed the protective effect of PBMT against muscle cachexia was totally blocked by AKT inhibitor in vitro and in vivo. Moreover, PBMT-activated AKT promoted FoxO3a phosphorylation and thus inhibiting the nucleus entry of FoxO3a. Lastly, in cisplatin-treated muscle cachexia model, PBMT had also been shown to ameliorate muscle atrophy through enhancing PI3K/AKT pathway to suppress MAFbx and MuRF-1 expression. These novel findings revealed that PBMT could be a promising therapeutic approach in treating muscle cachexia induced by cancer. Schematic representation of the signaling pathway for PBM therapy ameliorates cancer cachexia-associated muscle wasting. In cancer cachexia mice, myostatin and activin released by tumor cells act on the corresponding receptor ActRIIB by vascular transportation. Subsequent Smad2/3 are phosphorylated to reduce AKT activity and suppress FoxO3a phosphorylation. Dephosphorylated FoxO3a are translocated into the nucleus and induce the transcription of target genes (MAFbx and MuRF-1) which regulate the ubiquitin–proteasome systems, resulting in muscle wasting. After receiving PBM therapy, PI3K-AKT signaling is activated, and then stimulates protein synthesis by activating mTOR. mTOR activates the P70S6K, leading to protein synthesis. AKT also phosphorylates and inhibits the nucleus entry of FoxO3a, thereby alleviating muscle wasting.

HIF-1α plays a crucial part in hypoxia response by transcriptionally upregulating genes to adapt the hypoxic condition. HIF-1α is under severe cellular control as its exceptional activation is always associated with tumorigenesis and tumor progression. Here, we report L3MBTL3 serves as a novel negative regulator of HIF-1α. It is upregulated during hypoxia and acts as a transcriptional target of HIF-1α. In the nuclei, L3MBTL3 makes an interaction with HIF-1α and promotes its ubiquitination and degradation. These findings indicate L3MBTL3 forms a negative feedback loop with HIF-1α in vitro to dampen the hypoxic response.

Abstract Cancer cachexia-associated muscle wasting as a multifactorial wasting syndrome, is an important factor affecting the long-term survival rate of tumor patients. Photobiomodulation therapy (PBMT) has emerged as a promising tool to cure and prevent many diseases. However, the effect of PBMT on skeletal muscle atrophy during cancer progression has not been fully demonstrated yet. Here, we found PBMT alleviated the atrophy of myotube diameter induced by cancer cells in vitro, and prevented cancer-associated muscle atrophy in mice bearing tumor. Mechanistically, the alleviation of muscle wasting by PBMT was found to be involved in inhibiting E3 ubiquitin ligases MAFbx and MuRF-1. In addition, transcriptomic analysis using RNA-seq and GSEA revealed that PI3K/AKT pathway might be involved in PBMT-prevented muscle cachexia. Next, we showed the protective effect of PBMT against muscle cachexia was totally blocked by AKT inhibitor in vitro and in vivo. Moreover, PBMT-activated AKT promoted FoxO3a phosphorylation and thus inhibiting the nucleus entry of FoxO3a. Lastly, in cisplatin-treated muscle cachexia model, PBMT had also been shown to ameliorate muscle atrophy through enhancing PI3K/AKT pathway to suppress MAFbx and MuRF-1 expression. These novel findings revealed that PBMT could be a promising therapeutic approach in treating muscle cachexia induced by cancer.

LIGHT is a member of the TNF superfamily and a proinflammatory cytokine involved in liver pathogenesis. Many liver diseases involve activation of Toll-like receptor 3 (TLR3), which is activated by double-stranded RNA (dsRNA). However, the involvement of LIGHT in TLR3 implicated liver diseases is not clear. In this study, we investigated the role of LIGHT in TLR3 involved liver pathogenesis by using a mouse model of TLR3 agonist poly(I:C)-induced hepatitis. We found LIGHT expression at both protein and mRNA level in liver tissues is dramatically increased during the course of poly(I:C)-induced liver injury. This induction depends on NF-κB activation as pretreating the mice with a NF-κB inhibitor abrogates LIGHT upregulation. Importantly, blockade of the LIGHT signaling pathway with the recombinant LIGHT receptor HVEM protein ameliorates liver injury in poly(I:C)-induced hepatitis. Conclusions. These results indicate that LIGHT amplification by NF-κB plays a significant role in TLR3 involved hepatitis and points LIGHT to be a potential drug target for liver disease therapy.

Rheumatoid arthritis (RA) is an autoimmune and inflammatory disease characterized by synovial inflammation, joint swelling, and cartilage and bone destruction. Dracocephalum heterophyllum (DH) is a compound in traditional Chinese herbal medicine well known for its anti-inflammatory, antiviral and antioxidant activities. In the present work, the therapeutic effects of DH were investigated in collagen-induced arthritis. Arthritis severity was assessed by clinical score, X-ray, and histopathological features. Expression of inflammatory cytokines was detected by qPCR and ELISA whereas anti-type II collagen antibodies were determined by ELISA. DH treatment significantly alleviated clinical scores, synovial inflammation, joint swelling, and cartilage and bone destruction. DH also reduced the production of inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and interleukin-1β (IL-1β), and decreased the serum levels of anti-type II collagen specific IgG antibodies in collagen-induced arthritis. The therapeutic effects of DH in collagen-induced arthritis provide evidence that DH might be a potential therapeutic drug for rheumatoid arthritis.

Colorectal cancer (CRC) is considered to be a leading cause of cancer-related death. Centromere protein O (CENPO) can prevent the separation of sister chromatids and cell death after spindle injury. Nevertheless, the role of CENPO in CRC has not been reported. The expression level of CENPO in CRC was revealed by TCGA database and immunohistochemical (IHC) staining. Subsequently, the loss-of-function assays were performed to identified the role of CENPO in CRC in vitro and in vivo. Our data demonstrated that CENPO was highly expressed in CRC. The expression of CENPO was positively correlated with the deterioration of CRC. Moreover, CENPO knockdown inhibited the malignant phenotypes of CRC cells, which was characterized by slowed proliferation, cycle repression at G2, promotion of apoptosis, reduced migration and weakened tumorigenesis. Furthermore, CENPO knockdown downregulated the expression of N-cadherin, Vimentin, Snail, CCND1, PIK3CA and inhibited AKT phosphorylation in CRC cells. Moreover, the function of CENPO in regulating proliferation and apoptosis depended on p53. In summary, CENPO may play a promoting role in CRC through the epithelial mesenchymal transition (EMT) and PI3K/AKT signaling pathway, which can be regarded as a molecular therapeutic target for CRC.

Colorectal cancer (CRC) is one of the most common malignancies and most of the patients diagnosed with advanced CRC have unsatisfactory treatment effect and poor prognosis. The purpose of this study was to investigate the effect of CCNI2 on the development of CRC. In this sutdy, immunohistochemical staining was used to detect CCNI2 expression levels in clinical samples, meanwhile, the Kaplan-Meier survival analysis was conducted. Celigo cell counting assay was used for screening shCCNI2s. QPCR and WB were performed to verify knockdown efficiency of CCNI2. Cell proliferation, colony formation, cell cycle, apoptosis, and mechanism investigation of CCNI2 knockdown were investigated by MTT assay, colony formation assay, fluorescence-activated cell sorting, and human apoptosis antibody array, respectively. Otherwise, the mouse model of CCNI2 knockdown was also constructed. The results of immunohistochemical staining and qPCR indicated that CCNI2 had a high expression level in the CRC tissues and cell lines. Kaplan-Meier survival analysis manifested that the high expression of CCNI2 suggested poor prognosis. The expression of CCNI2 was significantly reduced by CCNI2-siRNAs, and the downregulated expression level of CCNI2 inhibited CRC cell proliferation and colony formation, arrested cell cycle in G2 phase, as well as promoted cell apoptosis. The various indexes of solid tumor in mice models indicated that CCNI2 knockdown could suppress the growth of CRC tumor. Based on the comprehensive analysis of the above results, CCNI2 was contributed to the progression of CRC and could serve as a prognostic marker for CRC.

At present, colorectal cancer (CRC) has become a serious threat to human health in the world. Dipeptidyl peptidase 3 (DPP3) is a zinc-dependent hydrolase that may be involved in several physiological processes. However, whether DPP3 affects the development and progression of CRC remains a mystery. This study is the first to demonstrate the role of DPP3 in CRC. Firstly, the results of immunohistochemistry analysis showed the upregulation of DPP3 in CRC tissues compared with normal tissues, which is statistically analyzed to be positively correlated with lymphatic metastasis, pathological stage, positive number of lymph nodes. Moreover, the high expression of DPP3 predicts poor prognosis in CRC patients. In addition, the results of cell dysfunction experiments clarified that the downregulation of DPP3 significantly inhibited cell proliferation, colony formation, cell migration, and promoted apoptosis in vitro. DPP3 depletion could induce cell apoptosis by upregulating the expression of BID, BIM, Caspase3, Caspase8, HSP60, p21, p27, p53, and SMAC. In addition, downregulation of DPP3 can reduce tumorigenicity of CRC cells in vivo. Furthermore, CDK1 is determined to be a downstream target of DPP3-mediated regulation of CRC by RNA-seq, qPCR, and WB. The interaction between DPP3 and CDK1 shows mutual regulation. Specifically, downregulation of DPP3 can accentuate the effects of CDK1 knockdown on the function of CRC cells. Overexpression of CDK1 alleviates the inhibitory effects of DPP3 knockdown in CRC cells. In summary, DPP3 has oncogene-like functions in the development and progression of CRC by targeting CDK1, which may be an effective molecular target for the prognosis and treatment of CRC.

目的:分析研究回肠造口术的方法、适应证及并发症的处理方法.方法:选择2005年8月至2015年8月中山大学孙逸仙纪念医院收治的137例行回肠造口术患者,分析其适应证和并发症,并总结回肠造口术的方法和并发症的处理.结果:预防性回肠造口52例,发生吻合口瘘后治疗性回肠造口85例.发生并发症18例(13.1％):肠造口缺血坏死1例、肠造口肠管狭窄3例、肠管外翻脱出2例、肠造口旁疝1例、肠造口出血4例、转流不完全2例、造口旁皮肤溃烂3例与肠管回缩2例.结论:回肠造口术是预防和治疗吻合口瘘的重要手段,预防和处理回肠造口的并发症极其关键.

目的 探讨在结肠癌细胞株中P53基因突变对KLF5基因功能及表达变化的影响.方法 通过对常见结肠癌细胞株的筛选,选取P53野生型及KLF5基因蛋白表达相对较低的细胞株作为研究对象,应用质粒构建过表达载体转染目的细胞,运用PCR及Western blot技术进行验证,再用细胞迁移、侵袭、增殖实验观察结肠癌细胞株中P53突变与否对KLF5基因功能及表达的影响.结果 通过应用Western blot技术对常见的7株结肠癌细胞株中KLF5基因蛋白表达以及应用Sanger测序对细胞株中P53突变情况的筛选,选取结肠癌细胞株RKO作为研究对象,在成功构建P53R175H过表达载体及KLF5干扰和过表达载体,并转染到RKO细胞株中,并行Western-blot验证后,运用细胞迁移、侵袭及增殖实验,发现在没有P53R175H突变的RKO结肠癌细胞株中,KLF5的过表达能抑制细胞的迁移、侵袭及增殖的能力,而敲除KLF5的表达后,可以发现细胞的迁移、侵袭及增殖能力明显增加;而在P53R175H突变后的结肠癌细胞株RKO中,KLF5的过表达则会导致RKO的迁移、侵袭及增殖能力明显增强,而敲除KLF5的表达后迁移、侵袭及增殖能力下降.同时也发现RKO细胞中P53的突变会导致KLF5的表达水平的下降.结论 在高表达P53突变的结肠癌细胞株RKO中,KLF5的蛋白表达水平下降,而且其基因功能则从抑癌向促癌方向转换.

OBJECTIVE:To evaluate the clinical application of carbon nanoparticles labeled lymph node staining in curative laparoscopic resection for colorectal carcinoma.METHODS:Sixty-five patients undergoing curative laparoscopic resection for colorectal carcinoma in the Sun Yat-sen Memorial Hospital between September 2011 and June 2013 were prospectively enrolled and randomly divided into label group (with carbon nanoparticles, n=34) and control group (without carbon nanoparticles, n=31). Association between labeled lymph nodes and metastasis was analyzed. The total number of retrieved lymph nodes and lymph nodes metastatic ratio were compared between the two groups.RESULTS:Mean number of retrieved lymph node of the label group was higher as compared to the control group (22.3±4.2 vs. 15.4±3.5, P<0.05). The total number of retrieved lymph node was 725 in the label group and 478 in the control group. Among them, lymph node < 5 mm accounted for 4.6% (33/725) in the label group, which was higher than 2.0% (10/478) (P=0.025) in the control group. The number of black stain label lymph node was 412, with black stain ratio 56.8% (412/725) in the label group. Metastatic ratio of black stain nodes was significantly higher than that of non-stain nodes [28.6% (118/412) vs. 19.5% (61/313), P=0.005].CONCLUSIONS:The technique of carbon nanoparticles labeled lymph node staining in curative laparoscopic resection for colorectal carcinoma is easy and effective, which can increase the retrieved number of lymph nodes, especially for nodes < 5 mm. The black stain lymph nodes indicate higher risk of metastasis.

Objective Analysis of the inguinal canal anatomical structures and their significance by intraoperative observation. Methods From June 2011 to May 2012,a total of 115 male patients(10 patients with bilateral hernias) whom were undergone open tension-free hernioplasty were involved in the observation. In open operation field, with high definition video equipment for recording and playback of the procedure, distribution of cremaster and nerves inside the inguinal canal and the relationship between spermatic cord and the inguinal canal rings were analyzed. Results The visible rates of the genital branch of genitofemoral nerve, ilioinguinal and iliohypogastric nerves were 90.4%, 98.4% and 38.4%, respectively. Cremaster muscle wasΩshape wrapping the spermatic cord. There were natural anatomic planes between spermatic cord and the inguinal ligament and inguinal falx. The spermatic cord was relatively fixed inside the inguinal canal by attachment of the inner and outer rings. Conclusion Ilioinguinal nerve is relatively constant in the inguinal canal, which theoretically could be safely cut off alike with the iliohypogastric nerve, as they are cutaneous branch in this position. The fixation of spermatic cord by the inguinal rings is a protective mechanism to prevent indirect herniaoccurs. The anatomical planes between the spermatic cord and the inguinal canal can be used of division.

PURPOSE:The purpose of this study was to report the imaging findings of malignant pancreatic solid pseudopapillary tumors (SPTs) with macroscopic venous tumor thrombi.METHODS:The clinical features and imaging findings of 4 cases of malignant pancreatic SPT with venous tumor thrombi were retrospectively reviewed.RESULTS:The tumor thrombi were located in the splenic vein (n = 3) or the main portal vein and the proximal splenic vein (n = 1). Venous thrombi were connected with the main pancreatic tumors and showed venous filling defects on computed tomography and magnetic resonance imaging. Tumor thrombi primarily consisting of necrotic component and/or hemorrhage displayed no enhancement after contrast injection (n = 3). The enhancement pattern of the tumor thrombi that consisted mainly of tumor nests was consistent with pancreatic SPT (n = 1), that is, a slight enhancement in the arterial phase and a progressive enhancement in the portal venous phase and the equilibrium phase. Venous tumor thrombi associated with hemorrhage were hyperintense on both T1-weighted and T2-weighted images.CONCLUSIONS:It is uncommon for pancreatic SPTs to spread by invading the venous system and forming macroscopic venous tumor thrombi.