Cold ischemia time in liver transplantation:An overview

The standard approach to organ preservation in liver transplantation is by static cold storage and the time between the cross-clamping of a graft in a donor and its reperfusion in the recipient is defined as cold ischemia time(CIT).This simple definition reveals a multifactorial time frame that depends on donor hepatectomy time,transit time,and recipient surgery time,and is one of the most important donor-related risk factors which may influence the graft and recipient’s survival.Recently,the growing demand for the use of marginal liver grafts has prompted scientific exploration to analyze ischemia time factors and develop different organ preservation strategies.This review details the CIT definition and analyzes its different factors.It also explores the most recent strategies developed to implement each timestamp of CIT and to protect the graft from ischemic injury.

Biliary tract injury caused by different relative warm ischemia time in liver transplantation in rats

BACKGROUND: There is a controversy over the degree of liver and biliary injury caused by the period of secondary warm ischemia. A liver autotransplantation model was adopted because it excludes the effects of infection and immunological rejection on bile duct injury. This study was undertaken to assess biliary tract injury caused by relative warm ischemia (secondary warm ischemia time in the biliary tract) and reperfusion. METHODS: One hundred and two rats were randomly divided into 5 groups: group I (control); groups 11 to V, relative warm ischemia times of 0 minute, 30 minutes, I hour and 2 hours. In addition to the levels of serum alkaline phosphatase, and total bilirubin, pathomorphology assessment and TUNEL assay were performed to evaluate biliary tract damage. RESULTS: Under the conditions that there were no significant differences in warm ischemia time, cold perfusion time and anhepatic phase, group comparisons showed statistically significant differences. The least injury occurred in group H (portal vein and hepatic artery reperfused simultaneously) but the most severe injury occurred in group V (biliary tract relative warm ischemia time 2 hours). CONCLUSIONS: Relative warm ischemia is one of the factors that result in bile duct injury, and the relationship between relative warm ischemia time the bile injury degree is time-dependent. Simultaneous arterial and portal reperfusion is the best choice to avoid the bile duct injury caused by relative warm ischemia. (Hepatobiliary Pancreat Dis Int 2009; 8: 247-254)

Warm ischemia time and elevated serum uric acid are associated with metabolic syndrome after liver transplantation with donation after cardiac death

AIM To describe the prevalence of posttransplant metabolic syndrome(PTMS) after donation after cardiac death(DCD) liver transplantation(LT) and the pre-and postoperative risk factors.METHODS One hundred and forty-seven subjects who underwent DCD LT from January 2012 to February 2016 were enrolled in this study. The demographics and the clinical characteristics of pre-and post-transplantation were collected for both recipients and donors. PTMS was defined according to the 2004 Adult Treatment Panel-Ⅲ criteria. All subjects were followed monthly for the initial 6 mo after discharge, and then, every 3 mo for 2 years. The subjects were followed every 6 mo or as required after 2 years post-LT.RESULTS The prevalence of PTMS after DCD donor orthotopic LT was 20/147(13.6%). Recipient's body mass index(P = 0.024), warm ischemia time(WIT)(P = 0.045), and posttransplant hyperuricemia(P = 0.001) were significantly associated with PTMS. The change in serum uric acid levels in PTMS patients was significantly higher than that in non-PTMS patients(P < 0.001). After the 1 s t mo, the level of serum uric acid of PTMS patients rose continually over a period, while it was unaltered in non-PTMS patients. After transplantation, the level of serum uric acid in PTMS patients was not associated with renal function.CONCLUSION PTMS could occur at early stage after DCD LT with growing morbidity with the passage of time. WIT and post-LT hyperuricemia are associated with the prevalence of PTMS. An increased serum uric acid level is highly associated with PTMS and could act as a serum marker in this disease.

Surgical Video Review of Warm Ischemia Time during Laparoscopic Partial Nephrectomy and Impact on Positive Surgical Margins and Postoperative Complications

Introduction: A surgical video review is an emerging tool for quality improvement, especially in complex surgeries such as laparoscopic partial nephrectomy (LPN). Assessing and measuring the warm ischemia time (WIT) during LPN by dividing it into the time used for resection (ResT), time used for reconstruction (RecT) and intermediate time (IntT) has not been performed before. This study aimed to analyze the factors that can influence all these surgical times and assess their impact on positive surgical margins (PSM) and complication rates. Methods: We evaluated 36 surgical video recordings from patients who underwent LPN and measured WIT, ResT, RecT and IntT with a stopwatch. Factors such as tumor characteristics and surgeon experience were also recorded. SPSS software was used to identify the predictor factors for all these surgical times and to correlate the ResT with PSM and RecT with the complication rate. Results: We recorded a mean WIT of 887 seconds. The mean ResT, RecT and IntT were 240 (27.2% of WIT), 473 (52.6% of WIT) and 173 s (20.2% of WIT), respectively. We found a moderate correlation between the WIT (p = 0.030), IntT and the R.E.N.A.L. score (p = 0.019). The surgeon with less than 100 LPN had significantly longer WIT, ResT, and RecT values, with means of 977 (p = 0.015), 268 (p = 0.019) and 530 seconds (p = 0.015), respectively. No correlation was found between ResT and PSM (p = 0.418);however, a strong correlation was found between RecT and the probability of developing complications (p = 0.012). Conclusion: The surgeon’s experience influences WIT, ResT, and RecT, but not IntT, which depends on tumor complexity. RecT affects the probability of developing complications. IntT represents a fifth of the WIT and efforts to reduce the WIT should focus on reducing the IntT for complex tumors, by improving surgical planning.

基于断指缺血时间与保存温度探讨影响断指再植效果的影响因素

目的分析在断指再植过程中,断指缺血时间以及保存温度对再植效果的影响。方法回顾性纳入商丘仁和中医院2021年1月至2023年12月期间收治的72例断指再植患者进行分析,收集患者的基本资料以及术后不良反应发生情况,收集术后2个月断指再植评估功能情况,并通过相关性分析明确断指缺血时间、保存温度与断指再植后功能的相关性。结果经评估,本次纳入研究的患者中,断指恢复情况优22例,良27例,差18例,劣5例;按照评估情况进行分组,四组患者断指再植功能评估各维度得分以及总分对比差异均有统计意义(P<0.05);四组患者的年龄、性别、受伤原因、受伤手指对比差异无统计学意义(P>0.05),而合并糖尿病、离断性质、离断平面、断指缺血时间以及保存温度比较,差异有统计学意义(P<0.05);优、良、差、劣四组患者术后不良反应的发生率分别为4.55%、7.41%、27.78%和100.00%,差异有统计学意义(P<0.05);Pearson相关性检验分析可知断指缺血时间、保存温度均与感觉恢复、日常生活活动、血液循环状态、恢复工作情况之间呈负相关,断指缺血时间与运动功能呈显著负相关关系(P<0.05);且除断指外观,其余功能恢复维度之间均呈正相关(P<0.05)。结论断指的缺血时间及保存温度与断指再植效果有关,尤其与断指再植后感觉恢复、日常生活活动、血液循环状态以及恢复工作情况之间密切相关,应重视加强相关科普宣传,改善患者术后恢复情况。

Real time RT-PCR定量检测BDNF mRNA表达水平方法的建立

目的建立real time逆转录聚合酶链反应(RT-PCR)检测BDNF mRNA基因表达的方法.方法提取脑缺血组织的总RNA,进行RT-PCR扩增BDNF mRNA特异性片段,扩增产物重组到质粒上并测序,建立real time RT-PCR检测BDNF mRNA表达水平方法.结果重组的质粒经酶切和测序,目的片段已插入到载体内,得到real time RT-PCR动力学曲线.结论成功建立real time RT-PCR检测BDNF mRNA基因表达的方法.

5-hydroxymethyl-2-furfural prolongs survival and inhibits oxidative stress in a mouse model of forebrain ischemia

In the present study, we hypothesized that 5-hydroxymethyl-2-furfural could attenuate ischemic brain damage by reducing oxidative injury. Thus, mice were subjected to bilateral common carotid artery occlusion to establish a model of permanent forebrain ischemia. The mice were intraperitoneally injected with 5-hydroxymethyl-2-furfura130 minutes before ischemia or 5 minutes after ischemia. The survival time of mice injected with 5-hydroxymethyl-2-furfural was longer compared with untreated mice. The mice subjected to ischemia for 30 minutes and reperfusion for 5 minutes were intraperitoneally injected with 5-hydroxymethyl-2-furfural 5 minutes prior to reperfusion, which increased superoxide dismutase content and reduced malondialdehyde content, similar to the effects of Edaravone, a hydroxyl radical scavenger used for the treatment of stroke. These findings indicate that intraperitoneal injection of 5-hydroxymethyl-2-furfural can prolong the survival of mice with permanent forebrain ischemia. This outcome may be mediated by its antioxidative effects.

Therapeutic window of Qingkailing injection for focal cerebral ischemia/reperfusion injury

The time window in which a drug is effective varies between drugs. The present study investigated the therapeutic window of Qingkailing injection for focal cerebral ischemia/reperfusion in mice. Animals underwent middle cerebral artery occlusion and were injected with Qingkailing （1.5, 3, 6 mL/kg）. Infarct volume and neurological function were assessed after 24 hours of ischemia. In addition, to establish the therapeutic time window, mice were injected with 3 mL/kg Qingkailing at 0, 1, 3, 4, 6, 9 and 12 hours after occlusion. Results revealed that Qingkailing injection significantly reduced infarct volume and improved neurological function in model mice after cerebral infarction for up to 9 hours, demonstrating that the therapeutic window of Qingkailing injection can extend to 9 hours for cerebral ischemia/reperfusion in mice.

To Optimize the Therapeutic Dose and Time Window of Picroside II in Cerebral Ischemic Injury in Rats by Orthogonal Test

The paper aims to optimize the therapeutic dose and time window of picroside II by orthogonal test in cerebral ischemic injury in rats. The forebrain ischemia models were established by bilateral common carotid artery occlusion (BCCAO) methods. The successful models were randomly divided into sixteen groups according to orthogonal experimental design and treated by injecting picroside II intraperitonenally at different ischemic time with different dose. The concentrations of neuron-specific enolase (NSE), neuroglial marker protein S100B and myelin basic protein (MBP) in serum were determined by enzyme linked immunosorbent assay to evaluate the therapeutic effect of picroside II in cerebral ischemic injury. The results indicated that best therapeutic time window and dose of picroside II in cerebral ischemic injury were ischemia 1.5 h with 20 mg/kg body weight according to the concentrations of NSE, S100B and MBP in serum. It is concluded that according to the principle of lowest therapeutic dose with longest time window, the optimized therapeutic dose and time window are injecting picroside II intraperitonenally with 20 mg/kg body weight at ischemia 1.5 h in cerebral ischemic injury in rats.

Assessment of Warm and Cold Ischemia on Functions of the Operated Kidney with ^99mTc-DMSA in Renal Masses: A Prospective and Randomized Study

Objective: To examine the effect of warm and cold ischemia on functions of the operated kidney in cases with a normal contralateral kidney undergoing nephron sparing surgery. Methods: This study enrolled 40 patients with a normal contralateral kidney and without a renal function threatening risk factor, who were operated with NSS. The patients were randomized at admission. They were divided into 2 equal groups as warm and cold ischemia. An ice application for 10 minutes was done to cold ischemia group after clamping renal artery. Renal functions were evaluated with Technesium-99m-Dimercaptosuccinic Acid (DMSA) and serum creatinine at the preoperative and postoperative (day 1, day 15, month 6, and month 12) period. Statistical analysis was done with Mann Whitney U test, Wilcoxon Signed Rank test, and Fredman test. A p value below 0.05 was considered statistically significant. Results: There were no significant differences between the groups in terms of age, body mass index, ischemia time, tumor size, amount of hemorrhage, and procedure time. Both groups had a significantly higher DMSA uptake at the preoperative period compared with the postoperative period (postoperative day 1, day 15, month 6, and month 12) (p 0.001). However, both groups had similar DMSA uptake results at the postoperative period. Preoperative and postoperative creatinine levels were not significantly different from each other in both groups. Conclusion: Based on tumor localization, nephron sparing surgery without use of superficial cooling appears as a viable option for small renal masses.

Protective mechanisms of picroside Ⅱ on aquaporin-4 expression in a rat model of cerebral ischemia/reperfusion injury

BACKGROUND： Aquaporin-4 （AQP-4） over-expression following cerebral ischemia results in cerebral edema. Picroside Ⅱ has been shown to exhibit a neuroprotective effect on neuronal apoptosis. However, few reports have addressed the neuroprotective mechanisms and therapeutic times following cerebral ischemic reperfusion injury. OBJECTIVE： To explore the neuroprotective effects and ideal treatment window for picroside Ⅱ treatment of middle cerebral artery occlusion and reperfusion injury in rats. DESIGN, TIME AND SETTING; A randomized, controlled, animal experiment was performed at Institute of Cerebrovascular Diseases, Qingdao University Medical College from September 2008 to May 2009. MATERIALS： Picroside II was purchased from Tianjin Kuiqing Medical Technology, China. METHODS： A total of 165 adult, healthy, male, Wistar rats were randomly assigned to sham-surgery （n = 15）, model （n = 75）, and treatment groups （n = 75）. Rats in the model and treatment groups underwent middle cerebral artery occlusion and reperfusion through the use of an intraluminal monofilament suture on the left external-internal carotid artery, The treatment group was injected with 1.0% picroside Ⅱ （10 mg/kg） into the tail vein, and the model and sham-surgery groups were injected with 0.1 mol/L phosphate buffered saline （250 μL）. MAIN OUTCOME MEASURES： Neurological functional scores were evaluated using the Longa＇s method; cerebral infarction volume was detected through the use of tetrazolium chlodde staining; cellular apoptosis was determined through the use of the in situ end-labeling method; aquaporin-4 expression was measured using fluorescence labeling analysis and reverse transcription polymerase chain reaction technique. RESULTS： At 0.5 hour following cerebral ischemic injury, neurological functional scores were low, and a small infarction focus was detected in the ischemic cortex of the model group. Along with prolonged ischemia and an increased number of apoptosis-positive cells, AQP-4 mRNA and protein expression was increased. At 1-2 hours after ischemia, neurological scores and infarction sizes were significantly increased in the model group. Apoptotic-positive cells were widespread in the ipsilateral cortex and stdatum. In addition, AQP-4 mRNA and protein expression levels were increased. Picroside II treatment significantly decreased neurological scores and infarction volume, and reduced AQP-4 mRNA and protein expression levels compared with the model group （P 〈 0.05 or P 〈 0.01）. At 1 hour after ischemia, the therapeutic effect of picroside Ⅱ was notable （P 〈 0.01）. CONCLUSION： Picroside Ⅱ played a protective role in cerebral ischemic reperfusion injury by inhibiting apoptosis and regulating AQP-4 expression. The best therapeutic time window was 1 hour after cerebral ischemic reperfusion.

The anti-inflammatory effect of picroside II and the optimizing of therapeutic dose and time window in cerebral ischemic injury in rats

The aim is to optimize the anti-inflammatory effect and the therapeutic dose and time window of picrosede II by orthogonal test in cerebral ischemic injury in rats. The forebrain ischemia models were established by bilateral common carotid artery occlusion (BCCAO) methods in 30 Wistar rats. The successful models were randomly divided into sixteen groups according to orthogonal experimental design and treated by injecting picroside II intraperitoneally at different ischemic time with different dose. The concentrations of aquaporins 4 (AQP4), matrix metalloproteinases9 (MMP9) and cyclooxygenase 2 (COX2) in serum and brain tissue were determined by enzyme linked immunosorbent assay to evaluate the therapeutic effect of picroside II in cerebral ischemic injury. The best therapeutic time window and dose of picroside II in cerebral ischemic injury were 1) ischemia 2.0 h with 20 mg/kg and 1.5 h with 20 mg/kg body weight according to the concentration of AQP4 in serum and brain tissue;2) ischemia 1.5 h with 20 mg/kg and ischemia 2.0 h with 20 mg/kg according to the concentrations of MMP9 in serum and brain tissue;and 3) ischemia 1.5 h with 10 mg/kg and ischemia 1.5 h with 20 mg/kg according to the concentrations of COX2 in serum and brain tissue respectively. According to the principle of the lowest therapeutic dose with the longest time window, the optimized therapeutic dose and time window were injecting picroside II intraperitoneally with 10 - 20 mg/kg body weight at ischemia 1.5 - 2.0 h in cerebral ischemic injury.

The Neuroprotective Effect of Picroside II and Its Best Therapeutic Dose and Time Window in Cerebral Ischemic Injury in Rats

Objective: To study the neuroprotective effect of picrosede II and explore the best therapeutic dose and time window according to orthogonal design in cerebral ischemic injury in rats. Methods: The forebrain ischemia rat models were established by bilateral common carotid artery occlusion (BCCAO) method. The successful models were randomly grouped according to orthogonal experimental design and treated by injecting picroside II intraperitoneally at different ischemic time with different doses. The contents of neuron-specific enolase (NSE), neuroglial marker protein S100B and myelin basic protein (MBP) in serum and brain tissue were determined by enzyme linked immunosorbent assay (ELISA) to evaluate the therapeutic effect of picroside II in cerebral ischemic injury. Results: The best therapeutic time window and dose of picroside II in cerebral ischemic injury may be 1) ischemia 1.5 h with 20 mg/kg and ischemia 1.5 h with 10 mg/kg body weight according to the content of NSE in serum and brain tissue respectively, 2) ischemia 1.5 h with 20 mg/kg according to the content of S100B in both serum and brain tissue, and 3) ischemia 1.5 h with 20 mg/kg and ischemia 1.5 h with 10 mg/kg according to the content of MBP in serum and brain tissue respectively. Conclusion: Based on the principle of the minimization of therapeutic drug dose and maximization of therapeutic time window, the optimal composition of the therapeutic dose and time window of picroside II in treating cerebral ischemic injury should be achieved by injecting picroside II intraperitoneally with 10-20 mg/kg body weight at ischemia 1.5 h in cerebral ischemic injury in rats.

Normothermic regional perfusion mobile teams in controlled donation after circulatory death pathway: Evidence and peculiarities

To facilitate the implementation of controlled donation after circulatory death(cDCD)programs even in hospitals not equipped with a local Extracorporeal Membrane Oxygenation(ECMO)team(Spokes),some countries and Italian Regions have launched a local cDCD network with a ECMO mobile team who move from Hub hospitals to Spokes for normothermic regional perfusion(NRP)implantation in the setting of a cDCD pathway.While ECMO teams have been clearly defined by the Extracorporeal Life Support Organization,regarding composition,responsibilities and training programs,no clear,widely accepted indications are to date available for NRP teams.Although existing NRP mobile networks were developed due to the urgent need to increase the number of cDCDs,there is now the necessity for transplantation medicine to identify the peculiarities and responsibility of a NRP team for all those centers launching a cDCD pathway.Thus,in the present manuscript we summarized the character-istics of an ECMO mobile team,highlighting similarities and differences with the NRP mobile team.We also assessed existing evidence on NRP teams with the goal of identifying the characteristic and essential features of an NRP mobile team for a cDCD program,especially for those centers who are starting the program.Differences were identified between the mobile ECMO team and NRP mobile team.The common essential feature for both mobile teams is high skills and experience to reduce complications and,in the case of cDCD,to reduce the total warm ischemic time.Dedicated training programs should be developed for the launch of de novo NRP teams.

肾部分切除术后急性肾损伤及其对手术侧肾脏的远期影响

肾部分切除术是治疗局限性小肾癌的首选方法,但手术过程中通常需要暂时阻断肾动脉,不可避免地引发缺血再灌注损伤,严重者甚至发生急性肾损伤。肾部分切除术后急性肾损伤的诊断常采用急性肾损伤网络标准(AKIN)和风险、损伤、失败、损失、终末期肾脏疾病标准(RIFLE),但使用总肌酐升高水平来评估单侧肾损伤显然不够准确;另外,肾部分切除术中会丢失部分正常肾组织,也导致血肌酐升高。为避免上述因素对诊断造成干扰,笔者首先改良了孤立肾肾部分切除术后急性肾损伤诊断分级方法,提出了诊断肾部分切除术后的急性肾损伤需要考虑到正常肾组织丢失引起的血肌酐升高。接着采用“极端值法”量化了非孤立肾患者手术侧肾脏急性肾损伤的程度。最后还探讨了急性肾损伤对手术侧肾脏的远期影响以及减轻肾部分切除术中缺血再灌注肾损伤的潜在方法。

Optimal therapeutic dose and time window of picroside II in cerebral ischemic injury

A preliminary study from our research group showed that picroside II inhibited neuronal apop- tosis in ischemic penumbra, reduced ischemic volume, and improved neurobehavioral function in rats with cerebral ischemia. The aim of the present study was to validate the neuroprotective effects of picroside II and optimize its therapeutic time window and dose in a rat model of cerebral ischemia. We found that picroside Ⅱ inhibited cell apoptosis and reduced the expression of neuron-specific enolase, a marker of neuronal damage, in rats after cerebral ischemic injury. The optimal treatment time after ischemic injury and dose were determined, respectively, as follows： （1） 2.0 hours and 10 mg/kg according to the results of toluidine blue staining; （2） 1.5 hours and 10 mg/kg according to early apoptotic ratio by flow cytometry; （3） 2.0 hours and 10 mg/kg according to immunohistochemical and western blot analysis; and （4） 1.5 hours and 10 mg/kg according to reverse transcription polymerase chain reaction. The present findings suggest that an intraperitoneal injection of 10 mg/kg picroside II 1.5-2.0 hours after cerebral ischemic injury in rats is the optimal dose and time for therapeutic benefit.

2-(2-Benzofuranyl)-2-imidazoline treatment within 5 hours after cerebral ischemia/reperfusion protects the brain

We previously demonstrated that administering 2-（2-benzofuranyl）-2-imidazolin（2-BFI）, an imidazoline I2 receptor agonist, immediately after ischemia onset can protect the brain from ischemic insult. However, immediate administration after stroke is difficult to realize in the clinic. Thus, the therapeutic time window of 2-BFI should be determined. Sprague-Dawley rats provided by Wenzhou Medical University in China received right middle cerebral artery occlusion for 120 minutes, and were treated with 2-BFI（3 mg/kg） through the caudal vein at 0, 1, 3, 5, 7, and 9 hours after reperfusion. Neurological function was assessed using the Longa＇s method. Infarct volume was measured by 2,3,5-triphenyltetrazolium chloride assay. Morphological changes in the cortical penumbra were observed by hematoxylin-eosin staining under transmission electron microscopy. The apoptosis levels in the ipsilateral cortex were examined with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling（TUNEL） assay. The protein expression of Bcl-2 and BAX was detected using immunohistochemistry. We found the following： Treatment with 2-BFI within 5 hours after reperfusion obviously improved neurological function. Administering 2-BFI within 9 hours after ischemia/reperfusion decreased infarct volume and alleviated apoptosis. 2-BFI administration at different time points after reperfusion alleviated the pathological damage of the ischemic penumbra and reduced the number of apoptotic neurons, but the protective effect was more obvious when administered within 5 hours. Administration of 2-BFI within 5 hours after reperfusion remarkably increased Bcl-2 expression and decreased BAX expression. To conclude, 2-BFI shows potent neuroprotective effects when administered within 5 hours after reperfusion, seemingly by up-regulating Bcl-2 and down-regulating BAX expression. The time window provided clinical potential for ischemic stroke by 2-BFI.

脑缺血/再灌注损伤后大鼠不同时间点神经细胞的动态变化

目的通过建立脑缺血/再灌注损伤模型,探讨急性CIRI后不同时间点神经细胞的动态变化规律。方法将SPF级雄性SD大鼠随机分为6组,分别为假手术组(Sham)和脑缺血/再灌注损伤(IR)不同时间点组。采用大脑中动脉线栓法(MCAO)建立局灶性脑缺血/再灌注损伤模型,采用Longa评分法评估大鼠神经行为评分,模型制备成功后,常规方法制作石蜡切片,进行TUNEL染色和免疫组织化学染色观察细胞凋亡,采用NeuN抗体进行免疫染色观察神经元细胞存活率,采用免疫组织化学法对星形胶质细胞标志物GFAP、小胶质细胞标志物IBA-1、内皮细胞标志物CD31进行染色,在不同时间点观察各组不同细胞的变化规律。结果假手术组的大鼠神经细胞在不同时间点未见明显变化,在脑缺血/再灌注损伤不同时间点组,细胞凋亡在IR3h被激活,且随着时间的延长,凋亡细胞数量增多且伴随形态学的破坏;NeuN+神经元在IR3h后出现缺血损伤的迹象,细胞形态异常,在24 h时最为严重;GFAP+星形胶质细胞在IR3h后急剧减少,IR6h标记不良星形胶质细胞数增加,到24、48 h星形胶质细胞在梗死区基本消失。IR3h IBA-1+小胶质细胞阳性细胞数减少,IR6h IBA-1+小胶质细胞体积增大,IR12h梗死区小胶质细胞死亡。CD31+内皮细胞在IR3h后在梗死周围皮层和纹状体明显增加,并持续到48 h。结论脑缺血/再灌注损伤后,凋亡细胞的数量随着时间延长而增加,NeuN+神经元细胞在24 h损伤最重;梗死周围皮层GFAP+星形胶质细胞、小胶质细胞随着时间增长而逐渐死亡;CD31+内皮细胞数量在再灌注3 h后在梗死周围皮层和纹状体明显增加,并持续到48 h。

基于3D Slicer软件的局限性肾癌供血动脉解剖分布研究

目的利用3D Slicer软件分析局限性肾癌供血动脉进入肿瘤的位置、数目与分布规律,为肾部分切除术中精准缝合提供解剖依据。方法收集2021年1月—2022年6月西安交通大学第二附属医院泌尿外科因肾癌行肾部分切除术的患者资料,将患者术前肾动脉计算机断层扫描(CT)血管成像资料以DICOM格式导入3D Slicer软件,从水平面、矢状面和冠状面对肿瘤-血管的相对位置进行重建,分析各平面中肿瘤供血动脉的数目及分布特点。结果共收集112例(男59例、女53例)肾癌患者相关资料,肿瘤均为单发,RENAL评分为4~10分,肿瘤分期T1a 58例、T1b 48例、T2a 6例。其中38例(33.93%)有1条肿瘤供血动脉、53例(47.32%)有2条肿瘤供血动脉、21例(18.75%)有3条肿瘤供血动脉。这207条肿瘤供血动脉中有22条(10.63%)经肿瘤-肾脏接触面(肿瘤床)浅部进入肿瘤,有185条(89.37%)经肿瘤床深部进入肿瘤。结论在局限性肾癌中,近90%的供血动脉由肿瘤床深部进入肿瘤,为肾部分切除术中精准肿瘤切除及创面缝合提供了解剖依据。

冷缺血时间对不同疾病类型肝组织质量的影响

目的 高质量的肝脏组织样本对肝脏疾病的研究至关重要,冷缺血时间(CIT)是影响组织样本质量的关键因素,探讨冷缺血时间与不同疾病类型肝组织样本质量的相关性,为获得高质量肝组织样本提供标准。方法 选取天津市第一中心医院进行手术治疗的肝癌(HCC)和胆道闭锁(BA)患者各30例,手术切除肝组织后,室温放置0、0.25、0.5、1、2、3、5和6 h后,分别从基因水平、蛋白水平及形态学水平对组织质量进行检测,提取总RNA及基因组DNA,分别检测浓度、纯度及完整性(RIN/DIN值),荧光定量PCR检测组织样本冷缺血0、6 h后ALB基因表达水平;免疫组化方法检测组织样本冷缺血0、6 h后Hep Par1、CK19、CD34、Vimentin蛋白表达情况,Westernblot检测组织样本冷缺血0、6h后ALB蛋白表达水平;HE染色观察组织样本冷缺血0、6 h后组织及细胞形态学变化。结果 DNA产量、RNA纯度和RNARIN随时间变化,时间效应具有统计学意义。组间效应不显著,表明肝癌和胆道闭锁组间没有差异。DNA产量和RNA RIN的交互效应有统计学意义,说明时间的影响随着组别不同而有所不同。与冷缺血0 h相比,肝癌、胆道闭锁组织样本冷缺血6 h后ALB基因表达水平降低,差异具有统计学意义(P <0.05);免疫组化方法检测组织样本冷缺血0、6h后Hep Par1、CK19、CD34、Vimentin蛋白表达情况均无变化,Western blot检测显示组织样本冷缺血0、6 h后ALB蛋白表达水平无显著变化;石蜡切片HE染色显示样本冷缺血0、6 h后组织及细胞形态学无变化。结论 冷缺血时间对肝组织样本中核酸质量具有显著影响,缩短冷缺血时间可获得高质量肝组织样本。