目的:探讨超声引导下经皮肝穿刺胆管取石术(PTCL)治疗肝移植术后胆道远期并发症的疗效及优势.方法:选择2例肝移植术后胆道远期并发症患者,运用超声引导下PTCL进行胆管取石、胆管狭窄扩张及胆道支架取出.分析2例患者的皮肤瘙痒改善程度、肝功能指标、感染指标及胆管狭窄变化.结果:2例患者治疗后皮肤瘙痒症状显著减轻,肝功能及感染指标显著降低,放置的引流管既能通畅引流又能支撑胆管狭窄.结论:超声引导下PTCL能有效解决肝移植术后胆道远期并发症,该方法安全可行.

Background: Ortner's syndrome, also known as cardio-vocal cord syndrome, is characterized by vocal cord paralysis secondary to compression of recurrent laryngeal nerve from a cardiopulmonary lesion. Clinicians may fail to diagnose or misdiagnose Ortner's syndrome because of its low incidence and complex pathogenesis. Here, we report a patient with an aortic arch aneurysm presenting solely with vocal cord paralysis and review existing literature to better understand Ortner's syndrome pathogenesis. Case presentation: On August 23, 2021, a 74-year-old man with cholecystolithiasis and acute cholecystitis was admitted to the Department of Hepatobiliary Surgery, at the First Affiliated Hospital of Jinan University. During medical history collection and physical examination, the patient's voice was unexpectedly hoarse. The patient then underwent laryngoscopy, computed tomography (CT) scanning of the chest, and enhanced CT scanning. Three-dimensional reconstruction of the thoracic aorta revealed an unstable aneurysm at the anterior of the aortic arch. The left recurrent laryngeal nerve was compressed by the aneurysm, leading to left vocal cord paralysis and voice hoarseness. Discussion: Hoarseness caused by vocal cord paralysis suggests damage to the recurrent laryngeal nerve. Examination based on the anatomical path, from the base of the skull to the diaphragm, especially from the vagus nerve origin area to the end of the recurrent laryngeal nerve, is helpful for early diagnosis of neck and thoracic cavity disorders, which could prevent misdiagnosis of acute and critical diseases.

恶性腹膜间皮瘤(malignant peritoneal mesothelio-ma,MPM)是一种来源于腹膜上皮或间皮组织的恶性肿瘤,几乎无敏感和特异的临床表现、实验室指标及影像学特征.诊断恶性腹膜间皮瘤的金标准是病理学及免疫组化,导致该病早期难以发现,且容易误诊,确诊时多为晚期[1].本文报道1例我院2019年10月收治的MPM患者,并对既往文献进行综述,旨在提高对MPM的认识,以便提高对MPM的诊治水平.

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are important proangiogenic factors in tumor procession. The autocrine and paracrine bFGF and the VEGF in tumor tissue can promote tumor angiogenesis, tumor growth, and metastasis. A VEGF/bFGF Complex Peptide (VBP3) was designed on the basis of epitope peptides from both VEGF and bFGF to elicit in vivo production of anti‐bFGF and anti‐VEGF antibodies. In this study, we reported on the production of recombinant VBP3 using high cell density fermentation. Fed‐batch fermentation for recombinant VBP3 production was conducted, and the production procedure was optimized in a 10‐L fermentor. The fraction of soluble VBP3 protein obtained reached 78% of total recombinant protein output under fed‐batch fermentation. Purified recombinant VBP3 could inhibit tumor cell proliferation in vitro and stimulate C57BL/6 mice to produce high titer anti‐VEGF and anti‐bFGF antibodies in vivo . A melanoma‐grafted mouse model and an immunohistochemistry assay showed that tumor growth and tumor angiogenesis were significantly inhibited in VBP3‐vaccinated mice. These results demonstrated that soluble recombinant VBP3 could be produced by large‐scale fermentation, and the product, with good immunogenicity, elicited production of high‐titer anti‐bFGF and anti‐VEGF antibodies, which could be used as a therapeutic tumor vaccine to inhibit tumor angiogenesis and tumor growth. © 2014 American Institute of Chemical Engineers Biotechnol. Prog ., 31:194–203, 2015

目的:通过检测ULK1和Beclin1在原发性肝细胞癌组织中的表达,结合数个临床病理因素,探讨ULK1和Beclin1在原发性肝细胞癌治疗中的临床意义.方法:手术取得80对肝癌和癌旁组织及20例正常肝脏组织,采用定量Real-time PCR(qPCR)及免疫组织化学技术检测各组织中ULK1和Beclin1的表达情况,联合临床病理因素进行统计分析.结果:ULK1和Beclin1在肝癌中蛋白表达及mRNA的表达,均低于在癌旁组织以及正常肝组织中的表达(P<0.05),而癌旁组织与正常肝组织中ULK1和Beclin1的表达的差异无统计学意义(P>0.05).ULK1和Beclin1在肝癌组织中的表达可能呈正相关关系(r=0.26,P=0.02).ULK1在肝癌中与性别、年龄、乙型肝炎表面抗原(HBsAg)、术前肝功能分级、甲胎蛋白(AFP)、嗜酒、肿瘤部位及分级无关,与肿瘤大小相关.Beclin1在肝癌中与性别、年龄、HBSAg、术前肝功能分级、嗜酒、肿瘤的部位及分级无关,但与肿瘤的大小及AFP表达相关.结论:ULK1和Beclin1可能通过影响自噬,进而肝癌的发生发展相关联.

1 病例资料 患者,男性,65岁,因"反复右上腹疼痛半月"入院.患者无明显诱因出现右上腹阵发性疼痛半个月,为阵发性绞痛,多于夜间发作,持续1～2小时后可自行缓解,无伴畏寒发热、黄疸、腹胀、恶心呕吐、呼吸困难等症状.行B超可见胆囊结石,结石大小约59 mm×15 mm.为求治疗,于2018年3月21日入院.既往史无特殊,个人史无特殊.

目的 探讨生长抑制因子2(ING2)和基质金属蛋白酶-13 (MMP-13)的表达与结直肠癌临床病理特征的关系.方法 应用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶法检测120例结直肠癌组织及40例癌旁正常结直肠黏膜组织中ING2和MMP-13蛋白的表达,并分析ING2和MMP-13蛋白的表达与结直肠癌临床病理特征的关系.结果 结直肠癌组织中ING2和MMP-13阳性表达率分别为71.66％ (86/120)和78.33％ (94/120),正常结直肠黏膜组织中ING2和MMP-13阳性表达率分别为25.00％ (10/40)和20.00％ (8/40),结直肠癌组织中ING2和MMP-13阳性表达率均显著高于正常结直肠黏膜组织(P＜0.05).结直肠癌组织ING2和MMP-13的表达与肿瘤组织分化程度、Duke分期、浸润深度及淋巴结转移有相关性(P＜0.05),但与患者的年龄、性别、肿瘤部位、肿瘤直径及远处转移无相关性(P＞0.05).Spearman等级相关性分析显示,结直肠癌组织中ING2与MMP-13表达呈显著正相关(P＜0.05).结论 ING2和MMP-13表达上调可能与结直肠癌的发生发展有关,ING2和MMP-13可作为判断结直肠癌恶性程度和预测预后的参考指标.

Fibroblast growth factor-2 (FGF-2) is one of the most important angiogenic factors to promote tumor growth, progression and metastasis. Neutralizing antibodies against FGF-2 may suppress the growth of tumor cells by blocking the FGF-2 signaling pathway. In this study, a disulfide-stabilized diabody (ds-Diabody) that specifically targets FGF-2 was designed. Compared to its parent antibody, the introduction of disulphide bonds in the diabody could significantly increase the stability of ds-Diabody and maintain its antigen binding activity. The ds-Diabody against FGF-2 could effectively inhibit the tube formation and migration of vascular endothelial cells and block the proliferation and invasion of human breast cancer cells. In the mouse model of breast cancer xenograft tumors, the ds-Diabody against FGF-2 could significantly inhibit the growth of tumor cells. Moreover, the densities of microvessels stained with CD31 and lymphatic vessels stained with LYVE1 in tumors showed a significant decrease following treatment with the ds-Diabody against FGF-2. Our data indicated that the ds-Diabody against FGF-2 could inhibit tumor angiogenesis, lymphangiogenesis and tumor growth.

PI3K-AKT-mTOR信号转导通路是哺乳动物肿瘤免疫中重要的信号通路,在多种恶性肿瘤的演变过程中发挥了极其重要的作用。近几年来,随着肿瘤分子生物学的发展,恶性肿瘤的靶向治疗成为研究热点,通过研究探讨PI3K-AKT-mTOR信号通路在肿瘤发生、发展过程中的信号转导机制,联合多种抑制剂或者寻找作用于多种信号通路、多靶点的新药,对于肿瘤的靶向治疗有重要意义。

The aim of the study was to investigate the possibility of human umbilical cord mesenchymal stem cells (UC-MSCs) surviving and differentiating into hepatocyte-like cells in partially hepatectomized model rats. MSCs were isolated from human umbilical cord and cultured with collagenase digestion. Cell surface markers were detected and fifth generation UC-MSCs were labeled with PKH26. The partially hepatectomized model rats were injected with the labeled human umbilical cord MSCs and transplanted through the portal vein. The survival of the labeled cells, in differentiation conditions and the expression of hepatic marker albumin were observed at post-transplantation 1, 2 and 3 weeks under a fluorescence microscope. It was found that the human umbilical cord MSCs could be cultured and amplified in vitro. Following transplantation to the partially hepatectomized liver of the model rat, the cells survived and expresses the hepatic marker albumin in vivo. After being labeled with PKH26, the cells were visualized as red fluorescence under a fluorescence microscope. In the frozen sections of the liver, the marked cells scattered around and most of them expressed albumin with green fluorescence under the fluorescence microscope. In conclusion, the transplanted human umbilical cord MSCs survived and differentiated into hepatocyte-like cells. The human umbilical cord MSCs may therefore be a main source of hepatocytes in transplantation.

1病例资料<br>　　患者，女性，76岁，因“发现右下腹肿块伴疼痛一周”入院，查体：右下腹膨隆，有压痛，无反跳痛，可触及一大小约4cm×4cm 包块，边界不清。结肠镜活检结果为乳头状管状腺癌，Ⅱ级；全腹 CT 提示：①右肾下级可见一肿块影，大小约3．8cm×3．5cm×3．9cm，考虑右肾癌；②升结肠肠管局部可见软组织肿块影凸向管腔，考虑升结肠癌。患者否认相关家族病史。常规行右肾癌根治术及右半结肠癌根治术。手术后病理报告：①中低分化腺癌，浸润至浆膜层，肠壁间淋巴结未见转移（0／12），肠系膜淋巴结未见转移（0／10）；②肾透明细胞癌，输尿管切缘未见癌累及，肾脂肪囊未查见肿大淋巴结。升结肠肿瘤免疫组化标记结果：癌细胞CD44（-），EGFR （-），VEGR （-），P53（＋＋），Ki-67约60％（＋）。右肾肿瘤免疫组化标记结果：癌细胞CK19（＋）， CD44（-），nm23（＋），VEGF （＋／-），Ki-67＜1％（＋）。诊断：（1）升结肠中-低分化腺癌（Dukes B）；（2）右肾透明细胞癌（T2N0M0）。患者术后恢复可，术后第五天恢复排气、排便，术后第九天伤口愈合拆线。

In the pathogenesis of asthma, central sensitization is suggested to be an important neural mechanism, and neurotrophins and cytokines are likely to be the major mediators in the neuroimmune communication pathways of asthma. However, their impact on the central nervous system in allergic asthma remains unclear. We hypothesize that central neurogenic inflammation develops in the pathogenesis of allergic asthma, and nerve growth factor (NGF) and leukemia inhibitory factor (LIF) are important mediators in its development. An asthma model of rats was established by sensitization and challenged with ovalbumin (OVA). For further confirmation of the role of LIF in neurogenic inflammation, a subgroup was pretreated with intraperitoneally (i.p.) LIF antibody before OVA challenge. The levels of LIF and NGF were measured with reverse transcription and polymerase chain reaction (RT-PCR), in situ hybridization (ISH) and immunohistochemistry stain in lung tissue, airway-specific dorsal root ganglia (DRG, C7-T5) and brain stem of asthmatic rats, anti-LIF pretreated rats and controls. A significantly increased number of LIF- and NGF-immunoreactive cells were detected in lung tissue, DRG and the brain stem of asthmatic rats. In the asthma group a significantly increase level of mRNA encoding LIF and NGF in lung tissue was detected, but not in DRG and the brain stem. Pretreatment with LIF antibody decreased the level of LIF and NGF in all tissues. LIF is an important mediator in the crosstalk between nerve and immune systems. Our study demonstrate that the increased level of LIF and NGF in DRG and brain stem may be not based on result from de novo synthesis, but rather on result from retrograde nerve transport or passage across the blood-brain-barrier.

Objective To compare the applied value of the pressure aggravation test and breath aggravation test in the diagnosis of early acute appendicitis. Methods A total of 101 cases with epigastralgia, middle or upper abdomen pain, disease duration within 6 hours undergoing pressure aggravation test and breath aggravation test respectively in our hospital between October 2010 and December 2012 were prospectively enrolled. By comparing with the postoperative pathological diagnosis (early acute appendicitis and other abdominal pain), the sensitivity and specificity of these two tests were calculated. Through analyzing the receiver operating characteristic (ROC) curve, the diagnostic value of early acute appendicitis was evaluated. Results Fifty-two cases of early acute appendicitis and 49 cases of other abdominal pain were diagnosed by postoperative pathologic results. The sensitivity and specificity of the pressure aggravation test were 87.5% and 72.1% and of the breath aggravation test were 53.8% and 83.7% respectively. The area under the ROC curve of the pressure aggravation test was 0.786 (95% CI: 0.693-0.878), similar to that of the breath aggravation test (0.688, 95% CI: 0.583-0.792). Conclusion The pressure aggravation test has higher value to diagnose early acute appendicitis, while the breath aggravation test has better specificity.

患者，男，63岁，因反复便血2个月，加重伴头晕、乏力2周，于2014年1月25日入院。患者2个月前无明显诱因解少量暗红色大便，平均每天约2次，伴有间断性腹胀，多见于进食后。2周前上述症状加重，血便量较多，每次约50 mL，2～3次／d，伴头晕、乏力，纳差。于是来我院就医，门诊查血常规：红细胞3.12×1012· L-1，血红蛋白82 g／L，红细胞压积27.7％；大便常规：红细胞0～2／HP，潜血试验（＋），诊断为消化道出血，收入院进一步诊治。起病以来，患者无寒战、发热、胸痛、呕血、咯血等，精神一般，睡眠、食欲较差，小便正常，体重减轻约6.5 kg。否认肝炎、结核病、糖尿病等病史，否认服用可疑药物史。

Aim:This study was to observe the differentiation of enhanced green fluorescent protein(EGFP)-labeled human umbilical cord mesenchymal stem cells(HUC-MSC) when implanted into liver-cut rat model.Methods: Human umbilical cord mesenchymal stem cells were isolated and analyzed of surface markers and then were transfected by EGFP-labeled lenti-viral vectors.SD rats were randomly divided into three groups: blank group,model group and experimental group.All the three groups were established liver-cut rat models and the blank group was not injected with HUC-MSC,the model group was injected with unlabeled HUC-MSC while the experimental group with EGFP-labeled HUC-MSC.Rat liver cells were isolated after 3 months and then for detection.The pathological structure was observed by frozen section HE staining and the CD90 expression was analyzed by flow cytometry while EGFP expression was observed by fluorescence microscopy.Results: The pathology revealed that the HUC-MSCs were partly gathered in intravascular emboli formation,which attached to blood vessels.The liver had inflammatory response after surgery in each group but the organization structure had no significant morphological differences in groups.The HUC-MSC surface markers were in high expression of CD44,CD29,CD105 and CD90,but low expression in CD34,CD45,CD106,CD31,CD40 and HLA-DR.The flow cytometry results showed GFP expression rate of the model group was 0.27%±0.21% and experimental group was 11.68%±1.46% with statistical significant(P<0.01).CD90 expression results showed in model group it was 0.843%±0.146% and in experimental group was(0.026%±0.017% with statistical significant(P<0.01).Conclusion:Human mesenchymal stem cells can differentiate stably into liver cells and are detected,but they are easily to form emboli.Liver cells have a low expression of CD90.

AIM: To investigate the role of p38 MAPK signaling pathway in the course of simvastatin-induced portal pressure(PP) reduction in the rats with cirrhosis and portal hypertension.METHODS: The rat model of cirrhosis and portal hypertension was induced by treating the animals with composite factors including carbon tetrachloride.These model rats were randomly divided into model group(n=10),simvastatin treatment group(n=11) and SB203580,treatment group(n=10).The rats in the latter 2 groups were treated with simvastatin and p38 MAPK inhibitor SB203580,respectively.Another eight normal rats served as normal controls.After the end of treatment,the PP of the model rats was measured and total p38 MAPK protein,phosphorylated p38 MAPK protein,total endothelial nitric oxide synthase(eNOS) protein,phosphorylated eNOS protein,and nitric oxide(NO) content in the livers were analyzed.RESULTS: The PP of the rats in model group was significantly higher than that in normal control group.The PP of the rats in both simvastatin treatment group and SB203580 treatment group was significantly lower than that in model group,and the PP of the rats in simvastatin treatment group was lower than that in SB203580 treatment group.No significant change of total p38 MAPK protein and total eNOS protein between normal group and model group was observed.Compared with normal control group,the expression levels of phosphorylated p38 MAPK protein increased and phosphorylated eNOS protein decreased in model group.Compared with model group,the expression levels of phosphorylated p38 MAPK protein decreased and phosphorylated eNOS protein increased in simvastatin treatment group and SB203580 treatment group.The increase in the expression level of phosphorylated eNOS protein in SB203580 treatment group was lower than that in simvastatin treatment group.The hepatic NO content in simvastatin treatment group and SB203580 treatment group was significantly higher than that in model group,and that in simvastatin treatment group was higher than that in SB203580 treatment group.CONCLUSION: Simvastatin lowers PP in the rats with cirrhosis and portal hypertension by inhibiting the activation of p38 MAPK signaling pathway.

BACKGROUND:Bioartificial liver could partial y replace the major liver functions, including detoxification, synthesis, secretion and biotransformation. OBJECTIVE:To use bibliometric indexes to track study focuses on bioartificial liver, and to investigate the relationships among geographic origin, impact factors, and highly cited articles indexed in Web of Science. METHODS:A list of citation classics for bioartificial liver was generated by searching the database of Web of Science-Expanded using the terms“artificial liver support system”or artificial liver or“bioartificial liver”. The top 33 cited research articles which were cited more than 100 times were retrieved. RESULTS AND CONCLUSIONS:Of 4 144 articles published, the 33 top-cited articles were published between 1992 and 2010. The highest citations paper was published in 2002, with a total of 668 citations, mean cited 55.67 per year. The total citations of 33 articles were 6 094 times, with a mean of 12.64 citations per article. These top-cited papers came from 11 countries, of which 12 articles came from the United States. University of Rostock led the list of classics with five papers. Harvard University and Massachusetts General Hospital ranked the second with four papers each. The 33 top-cited articles were published in 18 journals, predominantly Annals of Surgery and Hepatology, fol owed by Artificial Organs and Biotechnology and Bioengineering. Our bibliometric analysis provides a historical perspective on the progress of bioartificial liver research. Articles originating from outstanding institutions of the United States and published in high-impact journals are most likely to be cited.

目的:构建和表达二硫键稳定的抗碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)人源性双链抗体.方法:以抗bFGF人源性单链Fv抗体(single chain Fv,ScFv)基因为模板,以PCR和重叠PCR (overlap PCR)的方法,用短的连接肽连接抗体的轻重链可变区,并将其轻重链可变区的适当部位定点突变成半胱氨酸,构建成二硫键稳定的双链抗体(disulphide stabilized diabody,ds-Diabody).结果:测序证实ds-Diabod基因正确;Western blotting结果显示不同肽链间的二硫键能正确配对,2条肽链可成功形成二聚体;ELISA结果显示相对于SeFv,ds-Diabody能更好地与抗原bFGF进行特异性结合,并具有良好的稳定性.结论:成功构建和表达了抗bFGF人源性ds-Diabody,在ScFv的基础上改善了抗体的亲和力和稳定性,从而为bFGF抗体药物的研究奠定了基础.

According to their sources,liver stem cells can be divided into the liver-derived stem cells and non-liver-derived stem cells.Non-liver-derived liver stem cellsinclude bone marrow mesenchymal stem cells,adipose-derived mesenchymal stem cells,umbilical cord mesenchymal stem cells and umbilical cord blood mesenchymal stem cells,and umbilical cord blood mesenchymal stem cells,as one kind of the non-liver-derived liver stem cells,have the features of low immunogenicity,rich source,and can be amplified and differentiate into liver cells under the action of inducer,and then express hepatocyte-specific markers and related functions,which provide an experimental basis for the umbilical cord blood mesenchymal stem cells treating liver function failure.