Hepatocellular carcinoma (HCC) is the most common type of liver cancer in China, and high molecular heterogeneity of HCC has caused the limitations in clinical efficacy; therefore, it is of great importance to explore its heterogeneity based on molecular classification and develop individualized diagnosis and treatment strategies. Various high-throughput sequencing techniques and multi-omics techniques in recent years have helped to establish multiple molecular classification systems, which gives us a deeper understanding of the molecular heterogeneity of HCC. This article summarizes the molecular classifications of HCC and discusses their association with clinicopathological features. This article also analyzes the new targets for molecular targeted therapy and immunotherapy and proposes new ideas for precise diagnosis and individualized treatment of HCC.

Vascular interventional therapy for hepatocellular carcinoma (HCC) is the most important treatment modality for unresectable HCC, including transcatheter arterial chemoembolization and hepatic arterial infusion chemotherapy. With the development and application of new materials and surgical procedures, the above treatment regimens have been modified and optimized, and various clinical studies have discussed the clinical effect of different regimens in the treatment of HCC and have made new achievements, which provides new strategies and a theoretical basis for vascular interventional therapy for HCC in clinical practice. This article summarizes the new advances in the clinical research on vascular interventional therapy for HCC.

目的 探讨新型异喹啉类化合物12-4-Y[1-乙基-8-甲氧基-5苯基吡唑并(5,1-a)异喹啉]对大鼠肝星状细胞(HSC)凋亡的影响,并探讨其可能的机制.方法 以不同浓度(5、10、20、40和80 μg/ml)12-4-Y处理HSC-T6细胞,分别于孵育后12h、24h、36h和48h收集细胞,以MTT法检测细胞增殖,以Hoechst染色对凋亡定性,以流式细胞分析对凋亡定量,应用Western blot检测凋亡关键蛋白Caspase3表达变化.结果 12-4-Y处理12h可浓度依赖性(在5～80μg/ml范围内)抑制HSC-T6细胞增殖,细胞增殖率分别为(1.086 ±0.013)、(1.055 ±0.019)、(0.957±0.012)、(0.793±0.021)和(0.553±0.031),均显著高于对照组水平(1.215 ±0.014,P＜0.05);12-4-Y(20μg/ml)处理可时间依赖性抑制HSC-T6细胞增殖,在12 h、24 h、36 h、48 h,细胞增殖率分别为(0.957±0.012)、(0.852±0.024)、(0.641±0.032)和(0.492±0.017),与对照组(1.318±0.007,P＜0.01)差异具有显著性;12-4-Y(20μg/ml)处理24h时,Hoechst染色显示HSC-T6可见明显凋亡小体,流式细胞分析显示凋亡率为(29.23±1.20)％,显著高于对照组[(5.47±1.39)％,P＜0.001];蛋白质印迹显示凋亡关键蛋白Caspase3的裂解产物表达亦显著增加.结论 新型异喹啉类化合物12-4-Y可显著诱导活化HSC凋亡,其机制与激活凋亡关键蛋白Caspase3有关.

Objective To compare the safety and clinical efficacy between paclitaxel liposomes and paclitaxel in patients with advanced esophageal cancer .Methods A total of 90 patients with advanced esophageal cancer were enrolled into this study and were randomly divided into paclitaxel liposomes treatment group(treatment group) and paclitaxel treatment group(control group) . The patients of each group were treated with paclitaxel liposomes or paclitaxel 80 mg/m2 intravenously on day 1 and 8 ,and nedapla‐tin 75 mg/m2 intravenously on day 1 ,respectively .After two cycles of chemotherapy ,three weeks per period ,we evaluated the short term efficacy and adverse reactions according to the WHO standard .Results The short term efficacy between control group and treatment group showed no significance statistically (P>0 .05) .However ,the incidence of allergic reactions in the treatment group was significantly lower than control group (P<0 .05) .Conclusion Paclitaxel liposomal or paclitaxel combine with nedaplatin was effective equivalently in the treatment of patients with advanced esophageal cancer .But the allergic reactions of paclitaxel liposomal were lower than paclitaxel .It is worthy of clinical promotion .

DNA的胞嘧啶5-甲基化是一个重要的表观遗传学修饰,这种修饰在哺乳动物的发育、老化以及疾病中发挥重要作用.研究发现,TET蛋白家族可催化5-甲基胞嘧啶(5mc)羟基化的酶活性,从而生成5-羟甲基胞嘧啶(5hmc).TET蛋白的这种催亿作用可将DNA同一位点上的C、5mc、5hmc保持动态平衡.TET家族蛋白和5hmc与众多疾病的发生、发展有关,尤其是与肿瘤疾病的关系使人们对肿瘤的发生有了一个新的理解.

目的 研究过表达MORC2对人L02肝细胞脂肪变性程度的影响及可能机制.方法 利用慢病毒转染技术在L02肝细胞中过表达MORC2,qRT-PCR及Western blot鉴定过表达效果后,对L02肝细胞进行脂肪变性诱导,测定细胞甘油三酯(triglyceride,TG)含量并比较过表达前后L02肝细胞的脂肪蓄积程度;利用芯片技术对过表达MORC2前后的基因表达谱进行检测与Pathway分析,筛选过表达MORC2时发生变化的通路并选择对脂代谢有调控作用的p53进行研究;Western blot检测在L02肝细胞脂肪变性诱导时过表达MORC2对p53蛋白含量的影响;在过表达MORC2的基础上对L02肝细胞过表达p53,qRT-PCR及Western blot鉴定p53过表达效果后,通过TG测定观察过表达p53对过表达MORC2改变的L02肝细胞脂肪蓄积程度产生的影响.结果 qRT-PCR及Western blot证明过表达MORC2成功;过表达MORC2的L02肝细胞在脂肪变性诱导后TG含量较MORC2正常表达的L02肝细胞明显降低(P＜0.01);基因表达谱芯片及Pathway分析显示脂代谢调控相关的p53通路在过表达MORC2后发生变化;Western blot显示过表达MORC2后,L02肝细胞中p53蛋白水平在脂肪变性诱导前后均降低;qRT-PCR及Western blot验证p53过表达成功,且过表达p53可部分解除MORC2对L02肝细胞脂肪变性的缓解作用.结论 过表达MORC2可通过下调p53从而缓解肝细胞脂肪变性.

Objective To determine the effect of recombinant adenovirus p53 (rAd-p53) therapy in combi-nation with transcatheter hepatic arterial themoembolization (TACE) for patients with primary liver cancer (PLC). Methods A total of 72 patients with advanced PLC were enrolled in this study from Jan. 2008 to Dec. 2014. Patients were divided into TACE group (mitomycin 8 mg,epirubicin 5 mg/m2,oxaliplatin 30 mg/m2,n=36) and p53 treatment group (rAd-p53,mitomycin 8 mg,epirubicin 5mg/m2,oxaliplatin 30 mg/m2,n=36). The procedure was re-peated once every 28 days for 4 times as a course. The therapeutic effects were evaluated according to the clini-cal symptoms,CT scans and liver functions. Results In TACE group,the total effective rate (CR+PR) was 50%including no complete response (CR) and partial response(PR) of 50%,stable disease (SD) was 5.6%,and pro-gressive disease(PD) was 44.4%;In p53 treatment group,the total effective rate was 69.4%,including CR of 5.6%, PR of 63.8%,SD of 5.6%,and PD of 25%;The Karnofsky performance score in p53 treatment group was improved better than in TACE group(P<0.05);The adverse effects in rAd-p53-treated patients was transient and self-limited fever. Conclusion The rAd-p53 therapy in combination with TACE is effective for patients with advanced PLC.

中消化道恶性肿瘤少见,但近年来发病率逐渐上升,其临床表现多样,无明显特异性.本文对中消化道恶性肿瘤病理学特点、临床特征及影像学检查作一概述,旨在为中消化道恶性肿瘤的诊治提供更多的临床信息和帮助.

目的 评价重组人p53(rAd-p53)腺病毒注射液联合化疗治疗中晚期食管癌的疗效观察.方法 对30例食管癌患者进行了rAd-p53内镜下注射联合化疗(基因治疗联合化疗,GTCT组)与30例食管癌单纯化疗(单纯化疗,CT组)的对照临床观察.在GTCT组,每周1次行内镜下注射rAd-p53制剂1×1012 VP/支共4次,同时结合化疗,口服替吉奥胶囊50mg,口服,2次/d,d1-14×4.两组所用的化疗方案相同.结果 GTCT组和CT组治疗4、8、12周后有效率(完全缓解+部分缓解)分别63％、77％、80％和33％、37％、39％,差异有统计学意义(P＜0.01).GTCT组在治疗过程中有24例出现自限性发热,其他无明显不良反应.结论 rAd-p53联合化疗治疗食管癌有较好疗效,临床使用安全,不良反应轻微.

Objective To explore the efficacy of the gelatin sponge particle and embosphere microsphere embolization for the treatment of patients with hepatocellular carcinoma and tumor rupture. Methods 49 patients with rupture of hepatocellular carcinoma underwent transcatheter arterial embolization by gelatin sponge particle (n =23) or embosphere microsphere (n =26) embolization in recent 5 years,and instant hemostatic and rehaemorrhagia rates were compared. Results There was no difference between gelatin sponge particle and embosphere microsphere embolization on the effects of instant hemostatic (both were 100%);Seven days after intervention,the rehaemorrhagia rate in gelatin sponge particles was 13.0%(3/23),one patient was transferred to surgery,one patient was re-intervened and one patient dead because of haemorrhagic shock,respectively;The rehaemorrhagia rate in embosphere microspheres group was 3.8%(1/26,P<0.01),and the patient received re-intervention and the bleeding was stopped successfully. Conclusion The instant hemostatic effect is the same between the gelatin sponge particle and embosphere microspheres embolization in patients with rupture of hepatocellular carcinoma,however,the embosphere group has much less chance to rebleeding within 7 days.

目的 研究蛋白激酶Cδ亚型(protein kinase Cδ,PKCδ)与肝细胞脂肪变性、内质网应激的关系,探讨其在非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)发病机制中的作用.方法 用混合脂肪酸构建人正常肝细胞L02脂肪变性模型,油红O染色和甘油三酯(TG)试剂盒检测细胞脂变程度,实时荧光定量PCR检测PKCδ、Bip、XBP-1smRNA表达,Western blot检测其蛋白表达.通过siRNA瞬时转染技术沉默PKCδ在L02细胞中的表达后观察上述指标表达变化.结果 油酸和棕榈酸酯2∶1混合比例能够成功诱导肝细胞脂肪变性,与对照组单个细胞脂变面积(14.47±9.28)和TG含量(2.30±0.62) μ,g/mg相比,脂肪酸16 h组脂变面积(333.06±42.36)和TG含量(30.86±6.24) μg/mg均显著增加(P＜0.05).脂肪酸组肝细胞PKCδ、Bip,XBP-1s mRNA和蛋白表达水平较对照组显著升高(P＜0.05),具有时间依赖性.在相同脂肪酸诱导液处理的条件下,PKCδ siRNA转染组细胞脂变程度(30.92±1.29)％较对照组(55.32±6.58)％明显减轻(P＜0.05),且PKCδ siRNA转染组Bip、XBP-1s的表达较对照siRNA转染组明显降低.结论 PKCδ在NASH形成发展中发挥重要作用,沉默PKCδ能够通过抑制内质网应激减轻肝细胞脂肪变性程度.

目的 研究5-甲基胞嘧啶羟化酶TET1在肝脏再生时卵圆细胞向肝细胞分化过程中的作用.方法 采用SD大鼠2-乙酰氨基芴(2-AAF)灌胃加2/3肝切除法建立肝脏再生模型,胶原酶灌注联合percoll密度梯度离心法提取纯化大鼠原代卵圆细胞,免疫荧光实验鉴定卵圆细胞特异性标志物OV6的表达;RT-PCR和Western blot检测肝切后不同时间(3、6、9、12、15 d)TETl mRNA和蛋白表达变化.培养WB-F344卵圆细胞,SCF 20 Lg/L、HGF 10 μ,g,/L、EGF 10 μg/L、地塞米松1.0×10-7 mol/L和DMS0 1.5％处理7d,诱导其向肝细胞样细胞分化(对照细胞不加任何细胞因子).倒置相差显微镜观察卵圆细胞形态变化,RT-PCR和Western blot检测TET1表达变化.结果 ①成功建立肝再生动物模型,分离获得大鼠原代卵圆细胞,其OV6阳性率≥80％.②与对照细胞相比,卵圆细胞被诱导分化后,细胞形态由圆形变为梭形.③与对照细胞相比,卵圆细胞被诱导分化后,TET1 mRNA和蛋白水平均显著下降(P＜0.01),此时肝细胞特异性标记物ALB表达升高,卵圆细胞分化为肝细胞样细胞.结论 5-甲基胞嘧啶羟化酶TET1可能参与了卵圆细胞增殖和肝再生过程.

微小RNA(microRNAs,miRNAs)是一类内源性非编码小分子RNA,在肝纤维化形成中miRNAs可出现表达异常,这些异常表达的miRNAs不仅参与调控肝损伤修复反应,而且可作为评估肝功能储备的分子标志物.本文就miRNAs在肝纤维化发生发展中作用的研究进展作一综述.

目的 探讨肝细胞脂肪变性模型中Sec61α的表达及可能机制.方法 用软脂酸与油酸混合物诱导L02肝细胞发生脂肪变性,建立非酒精性脂肪性肝病肝细胞体外模型,按0、2、4、8、12h及16 h收获细胞;随后应用See61抑制剂Eeyarestatin Ⅰ处理肝细胞,分为正常对照组、脂肪变性组、DMSO组及抑制剂组,通过油红O染色、qRT-PCR及Western blot检测Sec61 α、GRP78、GRP94、及Bcl-2的表达变化.结果 软脂酸与油酸混合物成功诱导肝细胞发生脂肪变性.与对照细胞相比,脂肪变肝细胞Sec6lαmRNA相对表达量在脂肪变早期下降(2、4h,P＞0.05),晚期升高(8、12 h,P＜0.05);GRP78在8h开始升高,16 h达峰(P＜0.01).Sec61α与GRP78的蛋白水平与mRNA水平表达在时间梯度上变化一致.Eeyarestatin Ⅰ处理显著减轻肝细胞脂肪变程度;与脂肪变性组相比,抑制剂组肝细胞Sec61α、GRP78明显降低(P＜0.01),GRP94、Bcl-2则显著升高(P＜0.01).结论 Sec61α可能通过调节肝细胞内质网应激,参与肝细胞脂肪变的形成.

Objective To clarify the expression change and significance of phosphofurin acidic cluster sorting protein-2(PACS-2) in non-alcoholic fatty liver disease(NAFLD).Methods An in vitro NAFLD model was established by inducing the steatosis of HepG2 cells using palmitic acid.The cells were collected at the time points of 0,4,8,12,and 24 h after induction.The steatosis levels were measured by oil red staining.Rat NAFLD model was established in SD rats by feeding high fat diet,while the rats fed with normal food were set as control group.These rats were killed at 4,8,12,16 and 20 weeks after feeding.The expression of PACS-2,endoplasmic reticulum stress marker-glucose regulating protein 78(GRP78),Bax and Caspase-3 in the cell model and liver tissue samples were detected by q-PCR and Western blotting.Results The steatosis of HepG2 cells was induced by fatty acid successfully.The rat NAFLD model was established in SD rats successfully.The relative expression of PACS-2 at mRNA level was significantly decreased in the early stages(4 and 8 h),but increased in the late stages(12 and 24 h after treatment).That of GRP78 was up-regulated in 8 h,and then reached the highest level in 24 h after induction(P<0.01).That of Bax and Caspase-3 was also significantly increased in 12 and 24 h(P<0.01).The expression of these proteins showed similar trends as those at mRNA level(P<0.01).In the rat NAFLD model,the expression of PACS-2 protein was significantly decreased in the early stages(4 and 8 weeks),but then up-regulated in the late stages(16 and 20 weeks,P<0.01).The expression of GRP78 was up-regulated in 4 weeks,and reached the summit in 20 weeks(P<0.01).The expression of Bax and Caspase-3 were significantly up-regulated as compared the control in the late stage of NAFLD(P<0.01).Conclusion Mild expression of PACS-2 in the early stage of NAFLD might be involved in endoplasmic reticulum stress during the NAFLD process,and its up-regulation in the late stage may be associated with the apoptosis induced hepatocyte injury.

Objective To investigate whether hepatic stellate cells(HSCs) can differentiate to hepatocyte-like cells in vitro,and the expression and changes of some stem cell markers during the induction.Methods Primary rat HSCs were isolated by the standard collagenase perfusion and purified by density gradient centrifugation,and then identified with fluorescent immunocytochemistry for Desmin and GFAP.The HSCs were treated with the cytokines as bFGF(20 μg/L),FGF4(20 μg/L),HGF(20 μg/L) and IL-6(1 μg/L) for 7 d served as experimental group,while those cultured without any cytokine for 24 h served as control group.The morphological changes of HSCs were examined by inverted phase contrast microscopy,and the hepatocyte-like phenotypes were determined by detecting the mRNA and protein expression of AFP,ALB,and other stem cell markers expression by real time-PCR and immunocytochemical assay.Results Our primary culture method obtained a large number of primary rat HSCs,with a positive rate to Desmin and GFAP of over 95%.After a combined induction of bFGF,FGF4,HGF and IL-6 for 7 d,HSCs showed a dramatically increased expression of AFP and ALB(P<0.05) at mRNA and protein levels,with the shape changing from star-like to rotund or polygonal.But no such change was found in control cells.The combined induction also resulted in a significant decrease in mRNA expression of p75NTR(P<0.05) and CD90,c-kit,sox-2 and oct-4(P<0.01).The expression of these markers at protein level was also significantly decreased or even undetectable.Conclusion HSCs might be of the potential as stem cells to differentiate into hepatocyte-like cells in vitro.During the induction,the expression of some stem cell markers is significantly downregulated.

Question: A 61-year-old woman was referred for acid reflux, recurrent distention, and pain in the upper abdomen. She had a past history of cholecystolithiasis and laparoscopic cholecystectomy 3 years ago. Physical examination revealed no palpable superficial lymph node. Her abdomen was soft and nontender without abnormal mass. Gastroduodenoscopy demonstrated a subepithelial mass with intact overlying mucosa in the gastric fundus (Figure A). Endoscopic ultrasonography revealed a 14-mm, homogenous, and hypoechoic oval lesion in the muscular layer (Figure B). The mass was resected completely by endoscopic full-thickness resection, and the defect was securely clamped (Figure C). The tumor tissue was processed for pathologic examination, and histology is displayed in Figure D–F. What is your diagnosis of this gastric submucosal tumor? See the Gastroenterology web site (www.gastrojournal.org) for more information on submitting your favorite image to Clinical Challenges and Images in GI. Histology of the tumor revealed features of Castleman's disease (CD). Some lymphoid follicles showed features of hyaline vascular variant (HV) CD, presenting a ring-like accumulation of small lymphocytes that surround a regressive germinal center (Figure E), with vascularization (Figure G) and concentric follicular dendritic cell processes (Figure H). Infiltration of plasma cells were also noted in the interfollicular region (Figure F, I), indicating plasma cell variant (PV) CD. Therefore, the patient was diagnosed as mixed variant (MV) CD of the stomach. She recovered well postoperatively. Computed tomography of the chest and abdomen confirmed no lymph node enlargement. She was discharged without systemic chemotherapy. CD is a rare, non-neoplastic, lymphoproliferative disorder that exhibits as HV, PV, or MV histologically.1Cronin D.M. Warnke R.A. Castleman disease: an update on classification and the spectrum of associated lesions.Adv Anat Pathol. 2009; 16: 236-246Crossref PubMed Scopus (236) Google Scholar HV is usually unicentric, curable, and asymptomatic with a favorable prognosis. PV is more commonly multicentric, unresectable, and requires systemic therapy to reduce the risk of malignant progression.2Dispenzieri A. Armitage J.O. Loe M.J. et al.The clinical spectrum of Castleman's disease.Am J Hematol. 2012; 87: 997-1002Crossref PubMed Scopus (160) Google Scholar MV is considered to be a transitional type between HV and PV. MV could be unicentric or multicentric. Therapy for MV may differ according to the clinical manifestations.3Madan R. Chen J.H. Trotman-Dickenson B. et al.The spectrum of Castleman's disease: mimics, radiologic pathologic correlation and role of imaging in patient management.Eur J Radiol. 2012; 81: 123-131Abstract Full Text Full Text PDF PubMed Scopus (71) Google Scholar This case is noteworthy because MV of the gastrointestinal tract is extremely rare. Only 8 cases of gastrointestinal CD have been reported: 5 in the stomach, 2 in the duodenum, and 1 in the colon. To our knowledge, all of the stomach CD cases are HV, but the present case is the first documented gastric MV. This case highlights that CD should be considered in differential diagnosis of submucosal tumors of the stomach. Bin Wang and Jun Wang contributed equally to this work. Pathology review and images provided by Prof Hualiang Xiao, MD, PhD, Director, Department of Pathology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, China.

Objective To determine the expression of lysine-specific demethylase 1(LSD1) in esophageal squamous cell carcinoma(ESCC) and normal esophageal tissues,and study the relationship of LSD1 with clinical features,esophageal cancer cell proliferation and apoptosis.Methods The expression of LSD1 and proliferation related antigen(Ki67) in 86 ESCC tissue samples and 29 normal esophageal tissue samples were detected by immunohistochemical assay.TUNEL was used to detect the apoptosis of esophageal cancer cells.The relationship of LSD1 expression with clinical data was analyzed.Results The expression of LSD1 was significantly higher in ESCC than that in normal tissue(89.5% vs 51.7%,P<0.05).It was related to lymph node metastasis and tumor mass growth(P<0.05),but not to tumor cell apoptosis.The expression of LSD1 was correlated with the prognosis of the patients(P<0.05).Conclusion The high expression of LSD1 in ESCC is correlated with the prognosis of the patients,and may be regarded as a therapeutic target for ESCC.

<正>【据《Hepatology》2012年9月报道】题:针对氧化型低密度脂蛋白的特异免疫策略:减轻小鼠非酒精性脂肪性肝炎的一种新途径(作者BieghsV等)非酒精性脂肪性肝炎(NASH)的特征表现为肝内脂质积聚伴肝组织炎症,最终可进展至肝硬化。近来,Bieghs等证实造血干细胞中的清道夫受体(SRs)CD36及清道夫受体抗体(SR-A)缺失可减轻肝组织炎症。除参与摄取修饰脂蛋白外,CD36及其抗体还具有激活炎症反应等其他功能。

Objective To observe the healing state of tendon-bone interface in femoral tunnel after inlaid fixation of tendon-bone complex,and to evaluate its application effect.Methods Right anterior cruciate ligaments in 28 New Zealand rabbits were removed and reconstructed with autogeneic tendons of semitendinosus muscles.Specimens were collected at different time points after surgery,tension tests were performed and the histological status of tendon-bone interface healing in femoral tunnel was observed.Results The tension tests showed that there were 9 cases of tendon rupture and 1 case of pull-out of tendon-bone complex in postoperative 4 weeks.The rupture sites of reconstructed ligament all were in tendons after postoperative 8 weeks.The histological observation showed that the tendon-bone healing was the process including the forming of connection of fibrous tissue,growing of newly formed bone into tendon tissue and uninterrupted rebuilding of local connection of fibrous tissue.Conclusion The fixation effect of inlaid fixation of tendon-bone complex is reliable,which could promote the tendon-bone interface healing process effectively.