目的 分析儿童肝移植术后淋巴组织增殖性疾病(post-transplant lymphoproliferative disorder,PTLD)的临床及病理特点,为诊断及治疗提供参考依据.方法 收集2020年5月至2022年5月天津市第一中心医院儿童肝移植科收治的肝移植术后PTLD患者的临床及病理资料.包括性别、年龄、手术方式、术后免疫抑制方案、PTLD治疗方案、预后、临床表现、肝功能、血浆EBV-DNA、影像学检查结果,依据2016年WHO淋巴组织肿瘤分类进行病理学分型及免疫组化染色结果.回顾性分析患者的临床、病理及预后的特点.结果 肝移植术后经病理学确诊的PTLD患者8例,其中男性4例,女性4例,年龄为1～4岁.8例患者原发病均为胆道闭锁且均行亲属活体肝移植术.本组病例中,5例出现淋巴结肿大,5例出现消化系统症状(包括腹痛、肠梗阻、腹水、腹胀),4例出现肝功能异常,3例出现发热,1例出现肝、肾功能异常.血浆EBV-DNA平均为46072 copies/ml.本组病例中非破坏性、多形性和单形性分别占12.5%(1/8)、25%(2/8)和62.5%(5/8).5例单形性PTLD中伯基特(Burkitt)淋巴瘤、弥漫性大B细胞淋巴瘤和成熟T细胞淋巴瘤,分别占60%(3/5)、20%(1/5)和20%(1/8).患者确诊后均减量或停用他克莫司并联合利妥昔单抗治疗,其中6例接受化疗,2例接受血液透析治疗,2例行局部占位手术切除治疗.8例患者中7例病情缓解,1例死亡.结论 PTLD的早期诊断、根据病理分型选择合理的治疗方案有助于提高患者预后,接受化疗的患儿应警惕肿瘤溶解综合征并及时给予积极有效干预.

患者女性,40岁.因体检发现下咽肿物1个月入院.患者一般情况可,既往体健.否认高血压及糖尿病病史,无吸烟及饮酒史.自发病以来,无咳嗽、咳痰,无咽部异物感,无声音嘶哑,无呛咳,无进食饮水困难,无食欲减退及体质减轻等病史.专科检查:电子喉镜示左侧梨状窝处可见肿物,表面光滑,左侧披裂固定不动,双侧声带光滑,未见肿物(图1).

Background Tumor-associated antigens (TAAs) can be targeted in cancer therapy. We previously identified a monoclonal antibody (mAb) 12C7, which presented anti-tumor activity in lung cancer stem cells (LCSCs). Here, we aimed to identify the target antigen for 12C7 and confirm its role in LCSCs. Methods Immunofluorescence was used for antigen localization. After targeted antigen purification by electrophoresis and immunoblot, the antigen was identified by LC-MALDI-TOF/TOF mass spectrometry, immunofluorescence, and immunoprecipitation. The overexpression or silence of ENO1 was induced by lentiviral transduction. Self-renewal, growth, and invasion of LCSCs were evaluated by sphere formation, colony formation, and invasion assay, respectively. High-throughput transcriptome sequencing (RNA-seq) and bioinformatics analysis were performed to analyze downstream targets and pathways of targeted antigen. Results Targeted antigen showed a surface antigen expression pattern, and the 43–55 kDa protein band was identified as α-enolase (ENO1). Self-renewal, growth, and invasion abilities of LCSCs were remarkably inhibited by ENO1 downregulation, while enhanced by ENO1 upregulation. RNA-seq and bioinformatics analysis eventually screened 4 self-renewal-related and 6 invasion-related differentially expressed genes. GSEA analysis and qRT-PCR verified that ENO1 regulated self-renewal, invasion-related genes, and pathways. KEGG pathway analysis and immunoblot demonstrated that ENO1 inactivated AMPK pathway and activated mTOR pathway in LCSCs. Conclusions ENO1 is identified as a targeted antigen of mAb 12C7 and plays a pivotal role in facilitating self-renewal, growth, and invasion of LCSCs. These findings provide a potent therapeutic target for the stem cell therapy for lung cancer and have potential to improve the anti-tumor activity of 12C7.

Objective:To screen the differentially expressed exosomal miRNAs derived from liver cancer stem cells (LCSCs) and its effect on the malignant biological characteristics of liver cancer cells.Methods:miRNA expression profile chip was used to analyze the differentially expressed exosomal miRNA derived from LCSCs. The effects of miRNA on malignant phenotypes of LCSCs were identified. The cells were further treated with doxorubicin at different concentrations (0, 150, 300 μmol/L), and the expression level of miR-196a was detected by quantitative real-time PCR (qRT-PCR). The apoptosis of liver cancer cells cultured by exosomes derived from LCSCs (Exo-NC group) and exosomes derived from miR-196a inhibited LCSCs (Exo-Inhibitor group) and the activity of caspase3/7 under the action of exosomes from LCSCs were detected. Nude mice were randomly divided into Do-PBS group, Do-Exo-Inhibitor group and Do-Exo-NC group using random number table method, with 5 mice in each group, and the effect of miR-196a on nude mice xenograft tumor model with liver cancer cells was analyzed.Results:In this study, exosomes were isolated and purified from CD133 + Huh7 stem cell culture supernatant. miR-7162-3p, miR-1910-5, miR-3613-3p, miR-196a and miR-155-5p were up-regulated, while miR-1246 and miR-3613-5p were down-regulated. miR-7162-3p, miR-196a and miR-155-5p in exosomes had important effects on the self-renewal ability of LCSCs. miR-1910-5p, miR-196a and miR-155-5p had important effects on the invasion ability of liver cancer stem cells, among which miR-196a had the most significant inhibitory effect. Treatment for 24 h, the miR-196a expression level of the 0, 150 and 300 μmol/L doxorubicin was 0.96±0.05, 1.23±0.05 and 2.33±0.03 respectively, with a statistically significant difference ( F=996.90, P<0.001). Treatment for 48 h, the miR-196a expression level of the 0, 150 and 300 μmol/L doxorubicin were 1.02±0.07, 2.35±0.05 and 2.89±0.55 respectively, with a statistically significant difference ( F=303.00, P<0.001). When the concentration of doxorubicin was 0 and 300 μmol/L, the apoptosis rates of the Exo-NC group were 9.37%±0.19% and 11.64%±0.27%, and those of the Exo-Inhibitor group were were 18.80%±1.91% and 22.79%±1.57%, with statistically significant differences ( t=4.41, P=0.048; t=4.96, P=0.038). When doxorubicin was not used, the ratios of caspase3/7 in the Exo-NC group at 24 h and 48 h were 0.94±0.08 and 0.97±0.09, and those in the Exo-Inhibitor group were 1.56±0.01 and 1.58±0.01, with statistically significant differences ( t=11.41, P=0.008; t=6.07, P=0.026). Under 300 μmol/L doxorubicin, the ratios of caspase3/7 in the Exo-NC group at 24 h and 48 h were 0.95±0.07 and 1.36±0.08, and those in the Exo-Inhibitor group were 2.84±0.08 and 3.20±0.14, with statistically significant differences ( t=24.20, P=0.002; t=15.78, P=0.004). The results of xenograft tumor in nude mice showed that the tumor volumes of Do-PBS, Do-Exo-Inhibitor and Do-Exo-NC groups increased successively, which were (1 051.86±89.90) mm 3, (1 310.91±86.66) mm 3 and (2 185.14± 352.34) mm 3 respectively, with a statistically significant difference ( F=30.28, P<0.001). The weights of the transplanted tumors in the 3 groups increased successively, which were (0.36±0.10) g, (0.39±0.12) g and (0.76±0.16) g respectively, with a statistically significant difference ( F=11.81, P=0.002). The expression of miR-196a in tumors was significantly decreased after miR-196a inhibitor transfection. The expression levels of the 3 groups were 1.05±0.16, 0.38±0.08 and 2.17±0.26, with a statistically significant difference ( F=48.93, P<0.001). Conclusion:The exosomal secreted by LCSCs can enhance the resistance of liver cancer cells to doxorubicin by miR-196a.

[目的]探讨肝癌干细胞来源外泌体中差异表达的蛋白质,重点分析外泌体ANXA2分子对肝癌恶性生物学特性(自我更新、侵袭和转移能力)的影响.[方法]采用无血清悬浮培养法和干细胞标志物流式分选术分离人肝癌干细胞;分别采用干细胞球形成实验、体外侵袭转移能力鉴定分离所得肝癌干细胞;采用蛋白质谱分析肝癌干细胞来源外泌体中的差异表达蛋白质;siRNA靶向抑制ANXA2表达后,分别进行肝癌细胞球形成实验、体外侵袭与迁移实验检测外泌体ANXA2对人肝癌细胞自我更新、侵袭、转移能力的影响;收集肝癌患者和正常人血浆,通过凝胶排阻层析联合CD63免疫磁珠分离纯化外泌体,Western blot检测外泌体中ANXA2表达情况,结合临床病理资料,综合分析外泌体ANXA2的临床相关性.[结果]成功分离得到肝癌干细胞CD133+SK-Hep-1,其具有典型的肿瘤干细胞生物学特性.与CD133-SK-Hep-1细胞外泌体相比,CD133+ SK-Hep-1肝癌干细胞来源外泌体中具有多种差异表达蛋白,其中外泌体ANXA2表达明显上调.siRNA靶向抑制肝癌干细胞ANXA2表达后,其外泌体中ANXA2表达也下降.分别采用PBS、siRNA靶向抑制肝癌干细胞ANXA2后外泌体(Exo-LCSCs-siANXA2)、肿瘤干细胞来源外泌体(Exo-LCSCs)刺激肝癌细胞系HepG2、SNU398,其自我更新、侵袭和转移能力依次增强.Western blot结果显示,肝癌患者血浆外泌体ANXA2表达明显高于健康人血浆;肝癌患者血浆外泌体ANXA2表达与患者年龄、性别无明显相关性,而与肿瘤TNM分期和转移显著相关.[结论]高表达ANXA2的肝癌干细胞的外泌体对肝癌细胞恶性生物学特性具有调控作用.

Shadow will make power output loss for Building Integrated Photovoltaic system. Power output predicting of Building Integrated Photovoltaic is always complicated due to multi-parameters, especially under partial shading conditions. In this paper, effects of partial shadow on the photocurrent and series resistor is analyzed, relationship of shaded cell area proportion with photocurrent and series resistor is obtained respectively, and based on this, a simplified mathematical model to describe the relationship between shaded cell area proportion and current-voltage, power-voltage characteristics of a photovoltaic module under partial shading conditions is proposed and verified by experiments. Electrical performance of the photovoltaic module under different shading scenarios is predicted using the simplified mathematical model, and the predicted results have a high consistency with experiments using the monocrystalline silicon photovoltaic module. The results show that under same shaded area proportion, the shadow distributing in parallel with the short edge, which makes photovoltaic modules power output 3 W/m(2) under 10% shaded area, has much more negative effects than distributing in parallel with the long edge. The power output of a photovoltaic string protected by one bypass diode depends on the largest shaded cell area proportion, when the largest shaded cell area proportion is around 45%, the photovoltaic string has no contribution to power output of the whole photovoltaic module. Therefore, in order to minimize the influence of shadow on the power output of a photovoltaic module, for Building Integrated Photovoltaic system meeting inevitable shadow with regular shape, the photovoltaic module should be arranged to make the shadow distributed uniformly on the cells protected with one bypass diode; but for Building Integrated Photovoltaic system meeting inevitable shadow with irregular shape, the module of whom every piece of cells parallels a diode is recommended.

Background MCOLN1 (mucolipin subfamily, member 1) was first identified as an autophagic regulator, which was essential for efficient fusion of both autophagosomes and late endosomes with lysosomes. This study is aimed at investigating the role of MCOLN1 in the development of pancreatic ductal adenocarcinoma (PDAC). Methods Immunohistochemistry (IHC) assay was conducted to evaluate the expression level of MCOLN1 in 82 human PDAC tumor tissues. Overall survival (OS) and recurrence-free survival (RFS) analysis was performed to assess the prognosis of patients. Colony formation and MTT assays [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide] were performed to measure the proliferation capacity of tumor cells. The expression level of related genes was measured by RT-PCR (reverse transcription polymerase chain reaction) and western blot assays. The animal model was used to examine the effects of indicated protein on tumorigenesis in vivo. Results The results of IHC showed that a high level of MCOLN1 expression was associated with the poor clinical characteristics of PDAC patients. OS and RFS were significantly worse in patients with high MCOLN1 expression. Silencing of MCOLN1 dramatically blocked the proliferation of PDAC cells. Mechanism studies confirmed that knockdown of MCOLN1 decreased the expression of Ki67 and PCNA (proliferating cell nuclear antigen), two markers of cell proliferation. In vivo, MCOILN1 depletion reduced the formation and growth of tumors in mice. Conclusion The high level of MCOLN1 expression was associated with poor clinical outcomes of PDAC patients. MCOLN1 ablation could inhibit PDAC proliferation of both in vitro and in vivo, which provide a new insight and novel therapeutic target for the treatment of PDAC.

Increasing studies have demonstrated that most tumors consisted a subpopulation of cells with stem cell properties, known as cancer stem cells (CSCs). Accumulating evidence indicated that CSCs may be critical driving force for several types of cancer. Hence, it was necessary to develop therapeutic approaches specifically targeting CSCs. In this review, first, the biological properties of CSCs were introduced, including the self-renewal and differentiation, high tumorigenesis and invasiveness, resistance to chemotherapy and radiotherapy, genetic and epigenetic variations. Meanwhile, CSCs-targeted therapeutic strategies were summarized, including targeting cell surface markers, signaling pathways, CSC niches, differentiation therapy, and drug resistance for CSCs. Furthermore, clinical trials on anti-CSCs therapies supported the efficacy of these therapies, as well as their combination with conventional chemotherapy and radiotherapy. CSCs could be significantly eradicated, eventually resulting in inhibited tumor growth, metastasis, and recurrence. Thus, selectively targeting CSCs with various agents may be a novel and promising therapeutic strategy against cancer.

Cancer stem cells (CSCs) are a rare subset of cancer cells that play a significant role in cancer initiation, spreading, and recurrence. In this study, a subpopulation of lung cancer stem-like cells (LCSLCs) was identified from non-small cell lung carcinoma cell lines, SPCA-1 and A549, using serum-free suspension sphere-forming culture method. A monoclonal antibody library was constructed using immunized BLAB/c mice with the multipotent CSC cell line T3A-A3. Flow cytometry analysis showed that 33 mAbs targeted antigens can be enriched in sphere cells compared with the parental cells of SPCA-1 and A549 cell lines. Then, we performed functional antibody screening including sphere-forming inhibiting and invasion inhibiting assay. The results showed that two antibodies, 12C7 and 9B8, notably suppressed the self-renewal and invasion of LCSLCs. Fluorescence-activated cell sorting (FACs) found that the positive cells recognized by mAbs, 12C7 or 9B8, displayed features of LCSLCs. Interestingly, we found that these two antibodies recognized different subsets of cells and their combination effect was superior to the individual effect both in vitro and in vivo. Tissue microarrays were applied to detect the expression of the antigens targeted by these two antibodies. The positive expression of 12C7 and 9B8 targeted antigen was 84.4 and 82.5%, respectively, which was significantly higher than that in the non-tumor lung tissues. In conclusion, we screened two potential therapeutic antibodies that target different subsets of LCSLCs.

[目的]探讨热休克蛋白90α(HSP90α)和HSP90β在肝癌中的表达及意义.[方法]采用免疫组化法检测103例肝癌组织和79例非肝癌相关组织(正常肝组织,肝炎,肝硬化)中HSP90α和HSP90β的表达,并分析其与肝癌临床病理特征的关系.[结果]肝癌组织中,HSP90α和HSP90β的阳性表达率分别为82.52％和85.44％,均显著高于非癌组织(P＜0.05).HSP90α高表达与肝癌分化程度相关(P＜0.05),而与患者性别、病理类型、病理分期无关(P＞0.05).HSP90β高表达与肝癌分化程度和TNM分期相关(P＜0.05),而与患者性别、病理类型无关(P＞0.05).Spearman相关性分析显示,HSP90α和HSP90β在肝癌组织中协同表达(r=0.535,P＜0.001).[结论]HSP90α和HSP90β在肝癌组织中共同表达上调,可能可作为肝癌潜在的诊断和治疗靶点.