Molecular mechanisms of hepatic ischemia-reperfusion injury and preconditioning

Ischemia-reperfusion injury is, at least in part, responsible for the morbidity associated with liver surgery under total vascular exclusion or after liver transplantation. The pathophysiology of hepatic ischemia-reperfusion includes a number of mechanisms that contribute to various degrees in the overall injury. Some of the topics discussed in this review include cellular mechanisms of injury, formation of pro- and anti-inflammatory mediators, expression of adhesion molecules, and the role of oxidant stress during the inflammatory response. Furthermore, the roles of nitric oxide in preventing microcirculatory disturbances and as a substrate for peroxynitrite formation are reviewed. In addition, emerging mechanisms of protection by ischemic preconditioning are discussed. On the basis of current knowledge, preconditioning or pharmacological interventions that mimic these effects have the greatest potential to improve clinical outcome in liver surgery involving ischemic stress and reperfusion.     

Simplified nomenclature for compound flaps

The unique niche for compound flaps is their potential role for the repair of massive defects that demands the simultaneous restoration of multiple, missing tissue types. These complex flaps can be sorted into two major classes, and their subtypes on the basis of their means of vascularization are described. (1) Solitary vascularization, the composite flap: "multiple tissue components with a single vascular supply and dependent parts." (2) Combined flaps: (a) Siamese flaps: "multiple flap territories, dependent due to some common physical junction, yet each retaining their independent vascular supply"; (b) conjoint flaps: "multiple independent flaps, each with an independent vascular supply, but linked by a common indigenous source vessel"; and (c) sequential flaps: "multiple independent flaps, each with an independent vascular supply, and artificially linked by a microanastomosis." Many technical modifications that have improved or will improve the reliability of these flaps should not be confused as distinct flap types, but rather acknowledged as variations that can be more conveniently classified for the purposes of improved communication and research by using this basic schema as a guideline.     

腹腔镜肝部分切除术中肝血流阻断方法的选择及对患者肝功能的影响

目的:探讨三种不同的肝血流阻断方法用于腹腔镜肝部分切除术的安全性及有效性,为肝胆外科手术提供参考。方法:对我院2011年7月至2013年9月收治的83例肝脏肿瘤行肝部分切除术患者的临床资料进行分析。根据肝血流阻断方式,将所选病例分为三组:间歇性断流组(22例),完全断流组(28例),选择性断流组(33例)。观察并比较三组患者的手术时间、断流时间、术中出血量、输血量、术后肝功能指标的变化及并发症的发生率。结果:三组的平均手术时间和断流时间无显著差异(P0.05);选择性肝血流阻断组患者的术中出血量和输血量均显著少于间歇性断流组和完全断流组(P0.05);三种断流方式术后均发生并发症,但选择性阻断术后并发症的发生率明显低于间歇性断流和完全断流术,差异具有统计学意义(P0.05)。三组患者术后的血清总胆红素(TBIL)、丙氨酸转氨酶(ALT)及天冬氨酸转氨酶(AST)的水平均呈现不同程度降低,血清白蛋白(ALB)的含量升高;选择性断流组各指标变化更明显,数值更接近于正常水平(P0.05)。结论:与传统断流方法比,选择性肝血流阻断法能够减少剩余肝脏的再灌注损伤,有利于提高手术效果。但该方法存在一定局限,应根据患者实际情况选择断流方式。     

Intrahepatic Vascular Anatomy in Rats and Mice—Variations and Surgical Implications

Introduction

The intra-hepatic vascular anatomy in rodents, its variations and corresponding supplying and draining territories in respect to the lobar structure of the liver have not been described. We performed a detailed anatomical imaging study in rats and mice to allow for further refinement of experimental surgical approaches.

Methods

LEWIS-Rats and C57Bl/6N-Mice were subjected to ex-vivo imaging using μCT. The image data were used for semi-automated segmentation to extract the hepatic vascular tree as prerequisite for 3D visualization. The underlying vascular anatomy was reconstructed, analysed and used for determining hepatic vascular territories.

Results

The four major liver lobes have their own lobar portal supply and hepatic drainage territories. In contrast, the paracaval liver is supplied by various small branches from right and caudate portal veins and drains directly into the vena cava. Variations in hepatic vascular anatomy were observed in terms of branching pattern and distance of branches to each other. The portal vein anatomy is more variable than the hepatic vein anatomy. Surgically relevant variations were primarily observed in portal venous supply.

Conclusions

For the first time the key variations of intrahepatic vascular anatomy in mice and rats and their surgical implications were described. We showed that lobar borders of the liver do not always match vascular territorial borders. These findings are of importance for the design of new surgical procedures and for understanding eventual complications following hepatic surgery.     

Genetics and cardiovascular system: influence of human genetic variants on vascular function

Candidate gene association studies in cardiovascular diseases have provided evidence on the molecular basis of phenotypic differences between individuals. The comprehension of how inherited genetic variants are able to affect protein functions has increased the knowledge of how genes interact with environment in order to modulate a particular phenotype. Although it is known that the human genome contains more than 10 million SNPs, only a minor part of them are supposed to be functional. A causative SNP in a particular gene may confer a small to moderate effect in complex phenotypes, such as functions important to cardiovascular homeostasis. This paper is a selective review of the literature on the evidence for interactions between vascular function and naturally occurring genetic variants in endothelial nitric oxide synthase (eNOS) and beta-2 adrenergic receptor (ADRB2), two genes among those influencing vascular phenotype and examples for which there is a strong evidence base. eNOS and ADRB2 will be characterized, as well as the mechanisms by which the enzyme and the receptor work to control vascular responses will be described. Understanding the molecular mechanisms underlying gene-mediated vascular function and their modification by genetic variants is expected to result in a better comprehension about individual's phenotypic differences.     

Smear cytology in the intra-operative assessment of periodontoid pseudotumour of the craniocervical junction

OBJECTIVE: Periodontoid pseudotumours are lesions of presumed degenerative aetiology which typically occur in elderly patients with cervical myelopathy. The clinical and radiological differential diagnosis includes neoplastic and inflammatory lesions, and the exclusion of such lesions may be of value in the intra-operative management of the patient. This audit aimed to examine the value of intra-operative smear cytology in the management of this condition. METHODS: The intra-operative smear cytological features of four cases of periodontoid pseudotumour were reviewed. RESULTS: In each case the intra-operative cytological interpretation was in keeping with a non-neoplastic and non-inflammatory process. CONCLUSION: These results suggest that intra-operative smear cytology can provide information on the value in the intra-operative management of patients with this lesion.     

原发性肝癌合并下腔静脉癌栓的手术治疗

目的:探讨原发性肝癌合并下腔静脉癌栓的手术治疗方法.方法:采用肝切除加下腔静脉取栓治疗2例肝癌合并下腔静脉癌栓患者,取栓方法包括经荷栓肝静脉取栓(1例)和下腔静脉切开取栓(1例),又分在全肝血流阻断下取栓和在萨氏钳局部血管阻断下取栓.结果:2例肝癌及下腔静脉癌栓均得到成功切除,术中无明显并发症发生;术后无死亡;随访中一例存活11月;另1例已生存8个月.结论:肝癌合并下腔静脉癌栓的手术治疗安全可行,其基本术式为肝切除加下腔静脉切开取检.     

不同入肝血流阻断方式对荷瘤小鼠肝功能影响的实验研究

目的:探讨不同血流阻断方式对荷瘤小鼠肝细胞功能的影响。方法:选择昆明小鼠24只随机分为三组,正常对照组(Suspe-nded operation,SO)、肝门阻断组(Occlusion of the portal triad,OPT)、保留肝动脉持续阻断门静脉(Occlusion of portal vein,OPV)各8只。采用门静脉注射肿瘤的方法建立肝癌模型,建模后3天采用阻断范围为左外叶和中叶、阻断时间为60分钟的入肝血流阻断方式,复流后5天后,通过测量3组对肝脏的缺血再灌注损伤程度以及病理学变化来评价不同血流阻断方式对肝细胞功能影响的程度。结果:门静脉注射小鼠肝癌细胞8天后,对照组测量小鼠正常丙氨酸氨基转移酶(ALT)值为66.5±22.3 IU/L,OPT组值为276.3±80.5 IU/L,OPV组值为89.6±28.4 IU/L,两组比较有统计学差异(P0.01);对照组测量小鼠正常天冬氨酸氨基转移酶(AST)值为301.3±126.7 IU/L,OPT组值为1126.4±285.5 IU/L,OPV组值为438.6±150.7 IU/L,两组比较有统计学差异(P0.01),病理组织学OPV组肝细胞损伤程度明显较OPT组轻。结论:保留肝动脉持续阻断门静脉可以减轻荷瘤小鼠肝脏的缺血再灌注损伤。     

适用于磁压榨吻合的平移钳的设计

目的:提出了磁压榨吻合操作中可用于磁体夹持固定的平移钳的结构设计方案。方法:分析本团队多种原创性磁压榨吻合技术操作中存在的缺陷和不足,认为目前磁压榨吻合操作中依靠术者徒手控制或利用现有血管钳对磁体进行夹持和固定,是造成操作中诸多不便的主要原因。从力学角度分析了常规血管钳夹持磁体易导致磁体滑脱的原因,指出设计加工磁吻合专用钳是解决目前磁吻合操作不便的有效措施。在此基础上提出了利用平移钳来固定磁体的结构设计方案,并从力学角度进行了受力分析,设计出了用于磁压榨吻合操作中磁体夹持固定的平移钳。结果:平移钳的结构设计能够稳定夹持磁体,避免磁体滑脱;同时,借助齿轮传动结构控制钳头的平行移动能够更好地控制磁体的精准对吸和分离,可进一步简化操作,避免副损伤,节省手术操作时间。结论:平移钳能满足多种形状及大小的磁体的夹持和固定,可有效控制磁体的吸合与分离,极大地方便操作。该平移钳加工简单,使用方便,有助于推动磁压榨吻合技术在临床广泛开展。     

Hepatic neovascularization after partial portal vein ligation: novel mechanism of chronic regulation of blood flow

The present study was undertaken to investigate hepatic microcirculatory response following partial portal vein ligation (PPVL) in rats. Portal pressure was markedly increased 2-6 wk after PPVL, but no significant reduction in sinusoidal perfusion and hepatocellular injury were detected. However, marked neovascularization was observed in PPVL rats using intravital microscopy and scanning electron microscopy (SEM). Extremely high red blood cell velocity (2,000-4,900 microm/s) was seen in these vessels. Injection of fluorescein sodium via the carotid artery revealed that the neovessels originated from the hepatic arterial vasculature. This was further confirmed by clamping the common hepatic artery and phenylephrine injection from the carotid artery. These vessels maintained sufficient flow after massive sinusoidal shutdown elicited by the portal infusion of endothelin receptor B agonist IRL-1620. SEM also showed extensive neovascularization at the hilum. Additionally, clamping the portal vein decreased sinusoidal perfusion only by 9.5% in PPVL, whereas a 71.2% decrease was observed in sham. These results strongly suggest that the liver maintains its microcirculatory flow by vascular remodeling from the hepatic arterial vasculature following PPVL.     

P2 receptors in cardiovascular regulation and disease

The role of ATP as an extracellular signalling molecule is now well established and evidence is accumulating that ATP and other nucleotides (ADP, UTP and UDP) play important roles in cardiovascular physiology and pathophysiology, acting via P2X (ion channel) and P2Y (G protein-coupled) receptors. In this article we consider the dual role of ATP in regulation of vascular tone, released as a cotransmitter from sympathetic nerves or released in the vascular lumen in response to changes in blood flow and hypoxia. Further, purinergic long-term trophic and inflammatory signalling is described in cell proliferation, differentiation, migration and death in angiogenesis, vascular remodelling, restenosis and atherosclerosis. The effects on haemostasis and cardiac regulation is reviewed. The involvement of ATP in vascular diseases such as thrombosis, hypertension and diabetes will also be discussed, as well as various heart conditions. The purinergic system may be of similar importance as the sympathetic and renin-angiotensin-aldosterone systems in cardiovascular regulation and pathophysiology. The extracellular nucleotides and their cardiovascular P2 receptors are now entering the phase of clinical development. An erratum to this article can be found at     

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory rate and heart rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41±2.94 cmH₂O which was significantly higher (P<0.05) than before resection (6.87±2.39 cmH₂O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.     

The essential characteristics of a rating system

The principles of the Tanner-Whitehouse-Healy system for assessing skeletal maturity will be described and in particular the statistical basis for the scores given to each stage of each bone in the hand and wrist. The uses of the system, particularly in clinical cases, will be discussed together with reliability of the ratings. A revised system will be presented in which a part of the stages are amalgamated and the possibility of dropping some of the bones will be discussed. General discussion will be invited on the logical basis for selecting certain stages of certain bones as superior to others as assessors of maturity.     

Intermittent selective clamping improves rat liver regeneration by attenuating oxidative and endoplasmic reticulum stress

Intermittent clamping of the portal trial is an effective method to avoid excessive blood loss during hepatic resection, but this procedure may cause ischemic damage to liver. Intermittent selective clamping of the lobes to be resected may represent a good alternative as it exposes the remnant liver only to the reperfusion stress. We compared the effect of intermittent total or selective clamping on hepatocellular injury and liver regeneration. Entire hepatic lobes or only lobes to be resected were subjected twice to 10 min of ischemia followed by 5 min of reperfusion before hepatectomy. We provided evidence that the effect of intermittent clamping can be damaging or beneficial depending to its mode of application. Although transaminase levels were similar in all groups, intermittent total clamping impaired liver regeneration and increased apoptosis. In contrast, intermittent selective clamping improved liver protein secretion and hepatocyte proliferation when compared with standard hepatectomy. This beneficial effect was linked to better adenosine-5′-triphosphate (ATP) recovery, nitric oxide production, antioxidant activities and endoplasmic reticulum adaptation leading to limit mitochondrial damage and apoptosis. Interestingly, transient and early chaperone inductions resulted in a controlled activation of the unfolded protein response concomitantly to endothelial nitric oxide synthase, extracellular signal-regulated kinase-1/2 (ERK1/2) and p38 MAPK activation that favors liver regeneration. Endoplasmic reticulum stress is a central target through which intermittent selective clamping exerts its cytoprotective effect and improves liver regeneration. This procedure could be applied as a powerful protective modality in the field of living donor liver transplantation and liver surgery.     

Arterial blood ketone body ratio as an indicator of the no-return point in hepatic inflow occlusion without venous shunt in dogs

The effects of hepatic inflow occlusion without venous shunt on the viability of the liver were investigated with respect to liver energy metabolism in dogs, subjected to portal triad cross-clamping (Pringle's maneuver) for 10, 30 and 60 min. The concentrations of ketone bodies and the arterial blood ketone body ratio (KBR) were decreased markedly by hepatic inflow occlusion, but recovered upon recirculation. The initial velocity of KBR recovery was 0.150/min after 10-min clamping, 0.140/min after 30-min clamping and 0.032/min after 60-min clamping. KBR recovery was delayed when hepatic inflow occlusion exceeded 30 min, which indicates that hepatic inflow occlusion for 60 min causes severe inhibition of energy generation in liver mitochondria. These findings indicate that the safety period for hepatic inflow occlusion without venous shunt is between 30 and 60 min in dogs, and that mortality can be predicted by measuring the initial velocity of KBR recovery upon recirculation.     

Detection of cerebral embolism using transcranial Doppler sonography: can artifacts be reliably recognized?]

A requirement for the use of TCD for the detection of emboli in the field of cardiac and vascular surgery is the reliable differentiation between true emboli and artifacts. In ten healthy volunteers we carried out a study to establish the method with which artefacts can most reliably be identified. Automatic detection of increasing signal intensity misinterpreted 14% of all artifacts as emboli; 1.7% of all artifacts sounded suspicious for embolism, and 0.6% met the classical criteria of an embolus. Using simultaneous recording of the flow signal in two sections of the middle cerebral artery, all artifacts were identified on the basis of their simultaneous manifestation. Reliable intra-operative differentiation of emboli from artifacts requires attentive, continuous acoustic and visual analysis of signals by an experienced investigator familiar with the surgical procedure. The introduction of a multiple-depth algorithm might significantly improve the automatic detection program.     

Scanning electron microscope study of the liver microcirculation

During the experimental investigation performed in dogs and rats, by means of scanning electron microscopy of corrosive anatomical preparations, the spatial organization of all parts of the hepatic vascular bed (arterial, venous and lymphatic) has been studied, specific features of their components construction have been described. Within the limits of one hepatic lobule the number of vessels included in the portal vein system exceeds that of the arterial ones, originating from the proper hepatic artery system. In every part of the vascular bed the gradient of the form, orientation and pronouncement of the nuclei-containing zones in endotheliocytes and myocytes has been established. Various appliances participating in the blood and lymph stream regulation in different parts of the vascular bed have been revealed. As initial elements of the lymph bed, closed digital or loop-like capillaries should be regarded, they localize in the organ's connective tissue framework. Around the portal and hepatic veins and their branches, as well as around the biliary ducts, well developed plexuses of the lymphatic and blood capillaries and vessels localize, they are the main draining pathways of the organ. The degree of development and pronouncement of these plexuses depends on the lumen size in the formation they accompany.     

Isolation of human hepatocytes after hepatic warm and cold ischemia: a practical approach using University of Wisconsin solution

A method is described for isolating human hepatocytes from tissue fragments after warm and cold ischemia as experienced during hepatic resections. Cells with a high trypan blue dye exclusion and good culture characteristics were isolated by employing an initial tissue perfusion with UW solution. The method could facilitate transfer of liver tissues between distant centers for cell isolation studies.     

Hepatic fibrosis--role of hepatic stellate cell activation

Hepatic fibrosis is a reversible wound healing response characterized by accumulation of extracellular matrix (ECM), or "scar," that follows chronic but not self-limited liver disease. The ECM components in fibrotic liver are similar regardless of the underlying cause. Activation of hepatic stellate cells is the central event in hepatic fibrosis. These perisinusoidal cells orchestrate an array of changes including degradation of the normal ECM of liver, deposition of scar molecules, vascular and organ contraction, and release of cytokines. Not only is hepatic fibrosis reversible, but it is also increasingly clear that cirrhosis may be reversible as well. The exact stage at which fibrosis/cirrhosis becomes truly irreversible is not known. Antifibrotic therapies will soon be a clinical reality. Emerging therapies will be targeted to those patients with reversible disease. The paradigm of stellate cell activation provides an important framework for defining therapeutic targets.     

Regulation of surface antigens expression in Paramecium primaurelia: genetic and physiological factors involved in allelic exclusion.

Summary In Paramecium aurelia, allelic exclusion can be considered as a basic feature of the surface antigens system in the same way as intergenic exclusion. Our studies on allelic exclusion in G156/G168 heterozygotes show that (1) allelic exclusion does not depend on discrete regulatory genes dispersed throughout the genome; (2) it does not seem to be influenced by cytoplasmic factors; (3) it occurs regardless of the surface antigen expressed by the parental strains at the time of the cross.These results are discussed in relation to both intergenic and interallelic exclusion for which a common basis is proposed.