Background Immune exclusion inhibits antitumor immunity and response to immunotherapy, but its mechanisms remain poorly defined. In triple-negative breast cancer (TNBC), an aggressive and generally immune-rich subtype, an immune-cold microenvironment predicts poor prognosis due to a limited response to chemotherapy and immune checkpoint inhibitors. This study aimed to identify mechanisms regulating immune infiltration in TNBC. Methods We performed spatial transcriptomic analysis comparing immune-enriched versus immune-cold treatment-naïve TNBCs. Functional analyses, including loss-of-function and reconstitution experiments, were conducted to investigate the role of trophoblast cell-surface antigen 2 (TROP2), a key target of anticancer antibody drug conjugates (ADCs), in promoting TNBC progression. A humanized TROP2 syngeneic TNBC model was used to assess the effects of TROP2-targeting in combination with anti-programmed cell death protein 1 (PD-1) therapy. Additionally, data from patients treated with immune checkpoint blockade were used to test hypotheses from the preclinical findings. Results We reveal that TROP2 controls barrier-mediated immune exclusion in TNBC through claudin 7 association and tight junction regulation. TROP2 expression is inversely correlated with T-cell infiltration and predicts poor outcomes in TNBC. We demonstrate that TROP2 is sufficient to drive tumor progression in vivo in a CD8 T cell-dependent manner, while its loss deregulates expression and localization of multiple tight junction proteins, enabling T-cell infiltration. We show that TROP2 targeting via hRS7, the antibody component of the ADC sacituzumab govitecan, enhances the anti-PD-1 response and improves T-cell accessibility and effector function. Correspondingly, TROP2 expression is highly associated with lack of response to anti-PD-1 therapy in human breast cancer. Conclusions This study defines a new mechanism of barrier-mediated immune exclusion in cancer controlled by TROP2-dependent tight junctions. This mechanism drives tumor progression but can be targeted via TROP2-directed therapy to activate antitumor immunity and enhance immunotherapy response.

Background: Type 2 innate lymphoid cells (ILC2s) are essential for maintaining immune regulation and promoting tissue homeostasis in allergic asthma. How the development of gut microbiota on neonatal ILC2s influences allergic airway inflammation remains unclear. Here we focus on offspring ILC2 development in the context of alterations in maternal gut microbiota. Methods: C57BL/6 maternal mice were gavaged with OM-85 during pregnancy and/or lactation, ILC2-driven allergic airway inflammation in the OVA-sensitized adult offspring was observed. ILC2 development in offspring early life were investigated using recombinant (r)IL-33, rIL-25 and Bromodeoxyuridine in the vivo experiments. Further ILC2 promoting factors- IL-33 and IL-25 production in offspring early life were analysed. Finally, we examined the changes in gut microbiota and its metabolites in both dams and pups, and explored the effects of short-chain fatty acids (SCFAs) on IL-33 expression and secretion. Results: Maternal OM-85 administration restrained ILC2-driven allergic airway inflammation in the OVA-sensitized adult offspring. During ILC2 development in offspring early life, maternal OM-85 administration suppressed IL-33 and IL-25 production to inhibit ILC2 expansion and ILC2 responsiveness to alarmins, and infantile ILC2s could persist into adulthood. Maternal OM-85 administration increased SCFAs in breast milk and SCFA-producing gut probiotics (predominant Bacteroides and Blautia) in offspring, especially during pregnancy and lactation. SCFAs down-regulated IL-33 expression and reduced IL-33 secretion by inhibited gasdermin D (GSDMD) formation. Conclusion: Maternal OM-85 administration restrains ILC2-driven allergic airway inflammation in adult offspring by increasing offspring intestinal SCFAs to modulate ILC2 development at an early stage, demonstrating that the transgenerational effects of maternal OM-85 exposure on offspring innate immunity.

Abstract Background Interleukin‐33 (IL‐33) exacerbates asthma probably through type 2 innate lymphoid cells (ILC2s). Nevertheless, the association between eosinophilic asthma (EA) and ILC2s remains obscure, and the mechanisms by which IL‐33 affects ILC2s are yet to be clarified. Methods ILC2s were evaluated in peripheral blood mononuclear cells, induced sputum, and bronchoalveolar lavage fluid obtained from patients with EA. Confocal microscopy was performed to locate ILC2s in lung tissue and the mRNA expression of ILC2‐related genes was also evaluated in the EA model. The proliferation of ILC2s isolated from humans and mice was assessed following IL‐33 or anti‐IL‐33 stimulation. Results The counts, activation, and mRNA expression of relevant genes in ILC2s were higher in PBMCs and airways of patients with EA. In addition, ILC2 cell counts correlated with Asthma control test, blood eosinophil count, Fractional exhaled nitric oxide level, and predicted eosinophilic airway inflammation. IL‐33 induced stronger proliferation of ILC2s and increased their density around blood vessels in the lungs of mice with EA. Moreover, IL‐33 treatment increased the counts and activation of ILC2s and lung inflammatory scores, whereas anti‐IL‐33 antibody significantly reversed these effects in EA mice. Finally, IL‐33 enhanced PI3K and AKT protein expression in ILC2s, whereas inhibition of the PI3K/AKT pathway decreased IL‐5 and IL‐13 production by ILC2s in EA. Conclusions ILC2s, especially activated ILC2s, might be critical markers of EA. IL‐33 can induce and activate ILC2s in the lungs via the PI3K/AKT pathway in EA. Thus, using anti‐IL‐33 antibody could be a part of an effective treatment strategy for EA.

The poor efficacy of chimeric antigen receptor T-cell therapy (CAR T) for solid tumors is due to insufficient CAR T cell tumor infiltration, in vivo expansion, persistence, and effector function, as well as exhaustion, intrinsic target antigen heterogeneity or antigen loss of target cancer cells, and immunosuppressive tumor microenvironment (TME). Here we describe a broadly applicable nongenetic approach that simultaneously addresses the multiple challenges of CAR T as a therapy for solid tumors. The approach reprograms CAR T cells by exposing them to stressed target cancer cells which have been exposed to the cell stress inducer disulfiram (DSF) and copper (Cu)(DSF/Cu) plus ionizing irradiation (IR). The reprogrammed CAR T cells acquire early memory-like characteristics, potent cytotoxicity, enhanced in vivo expansion, persistence, and decreased exhaustion. Tumors stressed by DSF/Cu and IR also reprogram and reverse the immunosuppressive TME in humanized mice. The reprogrammed CAR T cells, derived from peripheral blood mononuclear cells of healthy donors or metastatic female breast cancer patients, induce robust, sustained memory and curative anti-solid tumor responses in multiple xenograft mouse models, establishing proof of concept for empowering CAR T by stressing tumor as a promising therapy for solid tumors.

目的:探究糖皮质激素对嗜酸粒细胞哮喘(Eosinophilic asthma,EA)2型固有免疫细胞(Type 2 innate lymphoid cells,ILC2s)的影响及相关机制.方法:研究对象来自我院2021年6月至2022年6月的EA患者和健康对照(Healthy control,HC),收集相应临床基线资料并评估病情、进行血常规、肺功能等检查;应用流式细胞术检测外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)ILC2s(CD45+Lin-CD127+CD294+);ELISA检测外周血IL-5、IL-13浓度.糖皮质激素治疗EA患者3月后,观察PBMC中ILC2s及 IL-5、IL-13浓度.C57BL/6J小鼠给予鸡卵清蛋白(Ovalbumin,OVA)20μg 腹腔注射致敏后用1％OVA雾化吸入激发哮喘EA模型,阴性对照(Negative control,NC)组小鼠用同等体积PBS作为对照.EA造模成功的小鼠通过流式细胞术检测血液及肺泡灌洗液中ILC2s,HE染色检测小鼠肺泡灌洗液中嗜酸性粒细胞(Eosinophil,EOS)及肺部炎症.EA小鼠经糖皮质激素处理后,检测肺部炎症情况;流式细胞术检测PBMC、肺泡灌洗液(Bronchoalveolar lavage fluid,BALF)中ILC2s;分离肺组织ILC2s,west-ern blot检测相关蛋白表达情况.结果:EA组的ILC2s比例升高,EOS升高,2型细胞因子 IL-5、IL-13增加,糖皮质激素治疗1月及3月后ILC2s比例下降,2型细胞因子IL-5、IL-13下降.与NC组小鼠比较,EA组小鼠PBMC及BALF中ILC2s升高,BALF中EOS升高,血清中2型细胞因子IL-5、IL-13升高,肺部炎症加重.糖皮质激素治疗后,肺部炎症减轻,EOS下降,ILC2s减少,2型细胞因子IL-5、IL-13下降,下调JAK/STAT蛋白.结论:在EA中,糖皮质激素通过下调JAK/STAT蛋白抑制ILC2s的功能减轻肺部炎症,为激素治疗嗜酸性粒细胞哮喘的机制提供了新方向.

BACKGROUND:There is a lack of information about regulatory T cells (Tregs) and inflammatory phenotypes in patients with asthma. In this study, we aimed to compare the characteristics of Tregs in patients with eosinophilic asthma.METHODS:Forty healthy and 120 stable asthmatic patients were recruited. Sputum and airway inflammatory phenotypes were assessed, and all patients were followed for one year. Human peripheral blood mononuclear cells (PBMCs) were collected and stimulated with phytohemagglutinin (PHA) and Dermatophagoides farina (Derp) to detect CD4+CD25+FOXP3+T cells and Foxp3 levels. Interleukin (IL)-13, IL-5, IL-17, IL-9, and interferon (IFN)-γ levels were measured.RESULTS:38.33% of patients had eosinophilic asthma, 13.33% had neutrophilic asthma, 6.67% had mixed granulocytic asthma, and 41.67% had pauci-granulocytic asthma. The eosinophilic asthma patients had a relatively high Asthma Control Test (ACT) score, an increased prediction and improvement FEV1 (%) rate, and elevated total IgE serum levels (P < 0.05). T helper cell 2 (Th2) cytokines IL-13 and IL-5 were predominantly expressed in the eosinophilic phenotype, while the Th1 cytokine IFN-γ and Th17 cytokine were found in the neutrophilic phenotype. IL-10 was significantly lower in eosinophilic asthmatic patients compared to the controls (P < 0.05). CD4+CD25+FOXP3+T cells (%Tregs) and Foxp3 gene expression in the PHA stimulated eosinophilic asthma samples were significantly lower compared to the control samples (P < 0.05). The airway inflammation phenotypes remained stable after one-year of therapy.CONCLUSION:Asthmatic patients with the eosinophilic phenotype in this study were deficient in Tregs, as characterized by a Th2 cell-biased pattern.

BACKGROUND:Deaths attributed to Coronavirus Disease 2019 (COVID-19) are mainly due to severe hypoxemic respiratory failure. Although the inflammatory storm has been considered the main pathogenesis of severe COVID-19, hypersensitivity may be another important mechanism involved in severe cases, which have a perfect response to corticosteroids (CS).METHOD:We detected the serum level of anti-SARS-CoV-2-spike S1 protein-specific IgE (SP-IgE) and anti-SARS-CoV-2 nucleocapsid protein-specific IgE (NP-IgE) in COVID-19. Correlation of levels of specific IgE and clinical severity were analysed. Pulmonary function test and bronchial provocation test were conducted in early convalescence of COVID-19. We also obtained histological samples via endoscopy to detect the evidence of mast cell activation.RESULT:The levels of serum SP-IgE and NP-IgE were significantly higher in severe cases, and were correlated with the total lung severity scores (TLSS) and the PaO2 /FiO2 ratio. Nucleocapsid protein could be detected in both airway and intestinal tissues, which was stained positive together with activated mast cells, binded with IgE. Airway hyperresponsiveness (AHR) exists in the early convalescence of COVID-19. After the application of CS in severe COVID-19, SP-IgE and NP-IgE decreased, but maintained at a high level.CONCLUSION:Hypersensitivity may be involved in severe COVID-19.

AbstractBackgroundCOVID‐19 is a highly contagious respiratory disease caused by the SARS‐CoV‐2 virus. Patients with severe disease have a high fatality rate and face a huge medical burden due to the need for invasive mechanical ventilation. Hypoxic respiratory failure is the major cause of death in these patients. There are currently no specific anti‐SARS‐CoV‐2 drugs, and the effect of corticosteroids is still controversial.MethodsThe clinical data of 102 COVID‐19 patients, including 27 patients with severe disease, were analyzed. The serum levels of total IgE and anti‐SARS‐CoV‐2 specific IgE were compared in healthy controls and COVID‐19 patients, changes in the level of anti‐SARS‐CoV‐2 specific IgE and clinical response to methylprednisolone (MP) treatment were analyzed, and the effect of high‐dose/short‐term MP therapy for patients with critical illness and respiratory failure was determined.ResultsCOVID‐19 patients had elevated serum levels of anti‐SARS‐CoV‐2 specific IgE, and patients with severe disease, especially critical illness, had even higher levels. Application of short‐term/high‐dose MP significantly reduced the level of these IgE antibodies and also blocked the progression of hypoxic respiratory failure. Hypoxic respiratory failure in patients with COVID‐19 is related to pulmonary hypersensitivity.ConclusionsHypersensitivity in the lungs is responsible for acute respiratory failure in COVID‐19 patients. Application of high‐dose/short‐term MP appears to be an effective life‐saving method for COVID‐19 patients who have hypoxic respiratory failure.

Structured abstract Aim: To elucidate the effect of miRNA (miR)-498 on autophagy and M2-like macrophage polarization in esophageal cancer. Methods: Autophagy was evaluated in esophageal cancer. Macrophage markers specific for M1- or M2-like phenotype were determined. The binding relationships between miR-498 and MDM2, MDM2 and ATF3 were analyzed. Results: miR-498 was downregulated in esophageal cancer and was associated with disease-free and overall patient survival. Enhanced miR-498 reduced LC3I conversion to LC3II and increased p62 accumulation in KYSE-150 cells, and increased macrophage polarization to M2-like phenotype in KYSE-150 and TAM co-culture. miR-498 inhibited MDM2-mediated ATF3 degradation, thus suppressing autophagy and M2-like polarization of macrophages in esophageal cancer. Conclusion: miR-498 may inhibit autophagy and M2-like polarization of macrophages to suppress esophageal cancer via MDM2/ATF3.

As the foremost common female malignancy, breast cancer (BC) poses a significant public health stumbling block. Although treatment protocols have improved over the years, the overall prognosis of BC remains unsatisfactory. Extensive investigations have taken place into long non coding RNAs (lncRNAs) pertaining to their involvement in carcinogenesis. The current study in connection with bioinformatics tools aimed to identify the myocardial infarction associated transcript (MIAT) as a BC-related differentially expressed lncRNA in an attempt to elucidate the effect of MIAT in BC cells. MIAT was initially overexpressed while DLG3 was down-regulated in BC. BC cells were subsequently treated with si-MIAT or/and si-DLG3, after which the expressions of DLG3 and the Hippo signaling pathway-related proteins were evaluated to analyze their regulatory mechanism in BC, which indicated that MIAT inhibition up-regulated DLG3 and activated the Hippo signaling pathway to suppress proliferation and promote apoptosis of BC cells. MS-PCR and RIP assays demonstrated that MIAT bound to the methylation proteins DNMT1, DNMT3A and DNMT3B, promoted the methylation of CpG islands in DLG3 promoter and inhibited the DLG3 expression. Moreover, our data suggested that DLG3 could bind to MST2 and regulate LAST1, which prevented the nuclear translocation of YAP. The in vitro results were further verified via the in vivo findings. Taken together, the central findings of our study demonstrate that MIAT silencing inhibits BC progression by means of up-regulating DLG3 via activation of the Hippo signaling pathway, highlighting a novel potential therapeutic target for the treatment of the BC.

目的:探讨影响老年弥漫大B细胞淋巴瘤治疗与预后的相关因素.方法:选取我院肿瘤科2008年7月至2014年7月收治的20例年龄大于65岁的弥漫大B细胞淋巴瘤病人,观察影响疗效及总生存率的各种变量,分析导致死亡的最主要因素.结果:16例病人接受治疗,4例病人未接受治疗.接受治疗的16例病人2年生存率为40％,中位生存时间为15个月.其中分期早、白蛋白高、IPI值2分以下、完成治疗的病人2年生存率高、预后好.利用COX比例风险回归模型分析感染、有无完成治疗是影响患者总生存期的因素.结论:感染是导致生存率下降的危险因素,也是导致死亡最主要的因素.

Background: Platinum compounds are commonly used for lung cancer treatment. However, the severe side effects and relatively poor prognosis limit their therapeutic effect. Therefore, developing novel platinum derivative and treatment strategy are critical for current lung cancer therapy. Methods: Flow cytometry, HMGB1 and ATP release, and immunoblotting were performed to evaluate the Oxaliplatin-induced immunogenic cell death (ICD) in two lung carcinoma cells. Vaccination approach and subcutaneous tumor models were created to analyze the tumor regression effect of Oxaliplatin. PD-L1 mRNA and protein levels were detected in LLC (Lewis lung carcinoma). Enhanced therapeutic efficacy of LLC was assessed by co-administration Oxaliplatin and aPD-L1 in murine lung tumor model. Results: Oxaliplatin induced robust ICD in LLC cells, activated dendritic cells (DCs, CD80(+)CD86(+)) and enhanced cytotoxic T cells (CD8(+)) in LLC tumor tissues, which resulted in tumor regression. Co-administration of Oxaliplatin and checkpoint inhibitor, aPD-L1, could enhance the therapeutic efficacy of LLC in murine lung carcinoma. Conclusion: This study reveals Oxaliplatin can induce robust ICD in tumor tissues and suppress tumor growth by activating DCs and enhancing T-cell infiltration. Notably, the Oxaliplatin-induced ICD provides an immunogenic microenvironment, which enhances the checkpoint inhibitor therapeutic efficacy of LLC.

Objective To discuss the efficacy and tolerance of three-dimensional conformal intensity-modulated radiation therapy (3D-CIMRT) for pulmonary metastasis and factors influencing prognosis.Methods Eighty-six patients with pulmonary metastasis treated at Nanchong Central Hospital from January 2014 to December 2015 were selected.All the patients underwent 3D-CIMRT.The short-term treatment effectiveness and patient's tolerance were analyzed.The 1-year and 2-year overall survival rates and progressfree survival rate were recorded and the Cox risk model was used for univariate and multivariate analyses of the prognosis for pulmonary metastasis after 3D-CIMRT.Results The overall short-term treatment efficacy was 82.6％.Some patients were observed to have adverse reactions including grade Ⅰ ～ Ⅱ acute radiation pneumonitis,chest pain,radiation dermatitis,leukopenia and anemia without acute or adverse treatment-related adverse reactions.The 1-year and 2-year overall survival rates were 74.4％ and 60.5％.The 1-year and 2-year overall progression-free survival rates were 43.0％ and 36.0％.Univariate analysis showed that the prognosis of patients with pulmonary metastasis was related to the Karnofsky performance status (KPS) score,the number of metastases,the dose of gross tumor volume (GTV) and the short-term treatment efficacy (P ＜ 0.05).Cox multivariate regression analysis showed that the KPS score,the dose of GTV and the short-term treatment efficacy were independent prognostic factors for the survival of patients with pulmonary metastasis (P ＜ 0.05).Conclusion 3D-CIMRT can improve the short-term treatment efficacy and prolong the survival time in patients with pulmonary metastasis and the adverse reactions were tolerable.The KPS score,the dose of GTV and short-term treatment efficacy are independent prognostic factors for the prognosis of patients after 3D-CIMRT.

目的 分析头颈部肿瘤患者放疗期间营养状况与生活质量现状及二者的相关性.方法 选取2016年6月-2017年1月收治的82例头颈部肿瘤患者进行研究,均采用问卷调查的方式进行,采用自行研究设计的一般情况调查表进行患者基本资料的调查,采用生活质量评价量表(CQLQ-C30)进行生活质量评估,采用主观整体营养状况评价量表(PG-SGA)进行营养状况评估.结果 放疗前、放疗中、放疗后82例患者A、B、C级PG-SGA评分比较差异有统计学意义(P<0.05).放疗后,患者在躯体功能、角色功能、认知功能、情绪功能、社会功能、总体健康状况方面评分低于放疗前,疲倦、恶心呕吐、疼痛、气促、失眠、食欲缺乏、经济困难方面评分高于放疗前(P<0.05).躯体功能、角色功能、认知功能、情绪功能、总体健康状况及疲倦、疼痛、失眠、食欲缺乏等症状方面与营养状况存在明显相关性(P<0.05);社会功能、恶心呕吐、气促、便秘、腹泻、经济困难与营养状况无明显相关性(P>0.05).结论 头颈部肿瘤患者在放疗期间的营养状态呈逐渐下降趋势,应密切关注患者的营养状况,采取相应的措施,纠正营养状态,以达到最佳治疗效果,提高生活质量.

目的:探讨分析妇科恶性肿瘤患者心理情况、治疗依从性及其影响因素.方法:本研究选取2015年5月—2016年9月我院收治的妇科恶性肿瘤患者240例作为恶性肿瘤组,同期选取我院进行健康体检的健康人群100例女性作为对照组,采用症状自评量表(SCL-90)对所有参与者心理状态进行评价,同时采用我院自行设计的患者治疗依从性调查表和治疗依从性影响因素调查表对患者治疗依从性及影响因素进行调查.结果:恶性肿瘤组的各项心理指标均显著高于健康对照组(P<0.05);本研究240例恶性肿瘤患者中治疗依从性良好的患者共162例,治疗依从性差患者共78例;不同医疗付费方式、文化程度、孕育情况、病理分期、化疗药物反应的患者治疗依从性比较差异显著(P<0.05);但不同年龄段及婚姻状况患者治疗依从性比较,差异不显著(P>0.05).结论:妇科恶性肿瘤患者普遍存在心理问题和治疗依从性差等情况,需对其进行对症处理,以期提高妇科恶性肿瘤患者的治疗效果及预后.

目的:探讨节律基因Cry1表达的CRY1蛋白在食管癌中的表达及其与临床意义.方法:应用免疫组织化学方法检测40例食管癌组织,40例癌旁组织中CRY1的表达.结果:40例食管癌组织中CRY1的弱阳/阳性表达(免疫组化染色强度评分大于等于2分)率为17.5％,而40例食管癌旁组织标本CRY1的阳性表达率为75％,两组表达的差异性具有统计学意义(P＜0.01).且表达强度与患者年龄、性别,肿瘤组织性质、侵及深度、淋巴结转移情况无明显相关性.结论:Cry1基因在正常组织中高表达,而在食管癌组织中低表达,可能作为一种抑癌基因影响肿瘤的发生发展.

Objective To evaluate the effect of 3 dimensional conformal radiation therapy for recurrence esophageal carcinoma after radical radiotherapy.Methods 60 esophageal carcinoma patients with local recurrence after their initial radiotherapy received retreatment with stereotactic radiotherapy.The acuate side effect,survival rates and immediate response were evaluated.Results All 60 patients completed there seconed course radiotherapy.The overall response rate was 78.2% with CR in 35 patients(58.3%),PR in 12 patients(20%),SD in 8 patients(13.3%) and PD in 5 patients(8.3%).The survival rate of patients within 6 months was 65%(39/60),and for 1,2 and 3 year were 31(51.6%),33.3%(20/60),16.7%(10/60),respectively.Conclusion 3 dimensional conformal radiation therapy is an alternative effective method for the in-patients with recurrence esophageal carcinoma after radical radiotherapy.

Objective To investigate the effects of circadian rhythm genes-hPer1 and hPer2 on radiosensibility of esophageal cancer cell lines Eca-109.Methods The circadian rhythm expression of hPer1 of esophageal cancer cell lines,Eca-109,was induced with dark-culture.The circadian rhythm expression of hPer2 of Eca-109 was induced with phorbol 12-myristate 13-acetate(PMA).The expression of hPer1 and hPer2 was detected with PT-PCR.Eca-109 cells were irradiated at peak(PG-hPer1,PG-hPer2) and nadir(NG-hPer1,NG-hPer2) of hPer1 and hPer2 with 6mV-X-ray respectively.The apoptosis rate of Eca-109 were evaluated with TUNEL and flow cytometry.Results The apoptosis rate of Eca-109 cells of PG-hPer1 and PG-hPer2 were lower than those of NG-hPer1 and NG-hPer2(P0.05).Conclusion hPer1 and hPer2 over-expression can decrease the radiosensibility of esophageal cancer cell lines Eca-109.