Mild hypothermia does not affect liver regeneration after partial hepatectomy in mice

Background: The use of mild hypothermia has been suggested to be therapeutically useful in treating acute liver failure. It is not known if hypothermia influences liver regeneration. Aim: To assess the effect of hypothermia on liver regeneration in mice. Methods: After partial (70%) hepatectomy (PHx), C57BL6/J mice were randomly assigned to either a hypothermic group or a normothermic group. Controlled mild hypothermia was maintained for up to 3 h after surgery. In addition, assessment of liver mass restitution was examined by studying the induction of key cell cycle proteins (cyclin A, D1 and E) and hepatocyte proliferation [assessment of proliferating cell nuclear antigen (PCNA) protein expression] by Western blotting and DNA synthesis by measuring 5‐bromo‐2‐deoxyuridine (BrdU) incorporation by immunohistochemical techniques 45 h after PHx. Results: Partial hepatectomy induced a vigorous proliferative response in the remnant livers of both groups of mice (normothermic and hypothermic groups), as evidenced by the induction of key cyclins, PCNA and incorporation of BrdU after PHx. The liver/body weight ratio and both cyclin and PCNA protein expression as well as BrdU incorporation did not differ between the regenerating livers of hypothermic and normothermic groups. Conclusion: Mild hypothermia does not influence liver regeneration in mice.     

Low preoperative platelet counts predict a high mortality after partial hepatectomy in patients with hepatocellular carcinoma

AIM: To assess the validity of our selection criteria for hepatectomy procedures based on indocyanine green disappearance rate (K_(ICG)), and to unveil the factors affecting posthepatectomy mortality in patients with hepatocellular carcinoma (HCC). METHODS: A retrospective analysis of 198 consecutive patients with HCC who underwent partial hepatectomies in the past 14 years was conducted. The selection criteria for hepatectomy procedures during the study period were K_(ICG)≥0.12 for hemihepatectomy, K_(ICG)≥0.10 for bisegmentectomy, K_(ICG)≥0.08 for monosegmentectomy, and K_(ICG)≥0.06 for nonanatomic hepatectomy. The hepatectomies were categorized into three types: major hepatectomy (hemihepatectomy or a more extensive procedure), bisegmentectomy, and limited hepatectomy, Univariate (Fisher's exact test) and multivariate (the logistic regression model) analyses were used. RESULTS: Postoperative mortality was 5% after major hepatectomy, 3% after bisegmentectomy, and 3% after limited hepatectomy. The three percentages were comparable (P=0.876). The platelet count of ≤10×10~4/μL was the strongest independent factor for postoperative mortality on univariate (P=0.001) and multivariate (risk ratio, 12.5; P=0.029) analyses. No patient with a platelet count of >7.3×10~4/μL died of postoperative morbidity, whereas 25% (6/24 patients) of patients with a platelet count of ≤7.3×10~4/μL died (P<0.001). CONCLUSION: The selection criteria for hepatectomy procedures based on K_(ICG) are generally considered valid, because of the acceptable morbidity and mortality with these criteria. The preoperative platelet count independently affects morbidity and mortality after hepatectomy, suggesting that a combination of K_(ICG) and platelet count would further reduce postoperative mortality.     

Changes in platelet count after cardiac surgery can effectively predict the development of pathogenic heparin-dependent antibodies

Cardiopulmonary bypass (CPB) induces the release of platelet factor 4 (PF4) and patients are at risk of heparin-induced thrombocytopenia (HIT). This study was aimed to determine whether an abnormal evolution in platelet count (PC) after CPB is predictive of the development of HIT antibodies. Two abnormal PC patterns were defined: pattern P1, characterized by a decrease in PC following previous correction of thrombocytopenia occurring during CPB, and pattern P2, defined as a persistent low PC in the days following CPB. PC was evaluated for 10 d in 305 consecutive patients before and after CPB. Serotonin release assay (SRA) was carried out between days 8 and 10 to detect pathogenic heparin-dependent antibodies. Moreover, antibodies to heparin-PF4 (H-PF4) complexes were assayed by enzyme-linked immunosorbent assay. PC evolution after CPB was normal in 300 patients although antibodies to H-PF4 were frequently present (53.4%). Changes in PC were abnormal in five patients with pattern P1 (n = 4) or P2 (n = 1). As SRA was positive in four of the five cases, the positive predictive value of abnormal PC pattern for pathogenic HIT antibodies was 80%. Careful follow-up of PC after CPB makes it possible to predict with high specificity (99%) for those patients who develop pathogenic HIT antibodies.     

Predictive factors for platelet increase after partial splenic embolization in liver cirrhosis patients

BACKGROUND AND AIM: Partial splenic embolization (PSE) is often performed for improving thrombocytopenia in cirrhotic patients. We investigated the largely unclear predictive factors for platelet increase at both 1 month and 1 year after PSE. METHODS: Aimed at increasing the platelet count, PSE was performed in 42 cirrhotic patients with thrombocytopenia (platelets < 80 x 10(4)/mL) caused by hypersplenism. The clinical data were analyzed to clarify the predictive factors for platelet increase at 1 month (n = 42) and 1 year (n = 38) after PSE. RESULTS: The mean splenic infarction ratio was 76.7% +/- 11.2%. The platelet count increased to 259% +/- 112% and 228% +/- 75% of the pretreatment values at 1 month and at 1 year after PSE, respectively. Stepwise multiple linear regression analysis showed that the infarcted splenic volume had a positive independent association with the increase in platelet count at both 1 month (P = 0.00004) and 1 year (P = 0.005) after PSE (increase in platelet count (x10(4)/mL): at 1 month = 0.752 + 0.018 x infarcted splenic volume (mL), R(2) = 0.344; at 1 year = 2.19 + 0.01 x infarcted splenic volume (mL), R(2) = 0.203). Receiver operating characteristic analysis yielded a cut-off value of 388 mL of infarcted splenic volume for achieving an increase of 5.0-8.0 x 10(4)/mL in platelet count at 1 year. CONCLUSIONS: PSE can reduce the platelet pool and induce an increase in platelet count. This increase is greatly dependent on the infarcted splenic volume.     

Lack of endothelial autophagy does not impair liver regeneration after partial hepatectomy in mice

Liver sinusoidal endothelial cells (LSEC) are key elements in regulating the liver response to injury and regeneration. While endothelial autophagy is essential to protect endothelial cells from injury-induced oxidative stress and fibrosis, its role in liver regeneration has not been elucidated. This study was intended to investigate the role of endothelial autophagy in liver regeneration in the context of partial hepatectomy (PHx). Analysis of autophagy levels in rat LSEC after PHx indicated a tendency to decrease activity the first 2 days after surgery. PHx performed in mice with impaired endothelial autophagy (Atg7^flox/flox;VE-Cadherin-Cre⁺) and their littermate controls showed no differences neither in liver-to-body weight ratio, histological analysis, hepatocyte proliferation nor vascular integrity during the first 7 days after PH and liver regeneration was completely achieved. Our results indicate that endothelial autophagy does not play an essential role in the coordination of the liver regeneration process after PHx.     

慢加急性肝衰竭患者血小板计数及其动态变化的临床意义

目的肝衰竭患者往往存在严重的血小板减少现象。对慢加急性肝衰竭(ACLF)患者血小板的动态变化进行监测分析,探讨血小板水平及其变化与患者预后的关系,并分析血小板减少的原因。方法收集2014年9月-2016年9月于首都医科大学附属北京佑安医院住院的54例ACLF患者的临床资料。血小板由全自动血常规分析仪检测获得,促血小板生成素(TPO)水平采用ELISA方法检测获得。正态分布的计量资料2组间比较采用t检验;非正态分布的计量资料2组间比较采用Mann-Whitney U检验;计数资料组间比较采用χ2检验。预后相关参数采用单因素及多因素logistic回归分析;应用受试者工作特征曲线(ROC曲线)分析血小板计数变化对肝衰竭的诊断价值。结果 ACLF患者出院时死亡18例,病死率为33.3%。ACLF患者基线血小板为80.0(36.0~334.0)×109/L,较正常值明显下降。出院前血小板下降幅度在死亡组显著大于存活组[(-43.4±58.9)×109/L vs(-11.5±29.1)×109/L,t=-2.827,P=0.041]。以血小板变化-27.5×109/L为cut-off值,血小板变化>-27.5×109/L和≤-27.5×109/L组病死率比较,差异有统计学意义(22.2%vs 72.2%,χ2=12.623,P<0.01)。单因素分析表明:血小板变化、PTA、Glu为患者预后的影响因素。多因素分析表明血小板变化为ACLF患者预后的独立影响因素,ROC曲线下面积为0.743。其中24例ACLF患者检测了血浆TPO水平,出院时死亡8例,病死率为33.3%。ACLF存活组与死亡组TPO水平分别为(91.8±39.8)pg/ml和(93.3±46.5)pg/ml,2组比较差异无统计学意义(t=0,P=0.938)。结论 ACLF患者基线血小板显著低于正常血小板的范围,血小板下降幅度是ACLF患者出院时死亡的独立影响因素,血小板减少与TPO水平下降无关。     

Factors associated with liver cancer prognosis after hepatectomy: A retrospective cohort study

This article was to investigate risk factors influencing liver cancer prognosis after hepatectomy.Patients undergoing hepatectomy after being diagnosed with liver cancer in Zhongshan Hospital Affiliated to Xiamen University were collected in the retrospective cohort study between January 2012 and December 2017, and divided into disease progression and non-progression groups based on their prognostic status. Univariate analysis was performed on the patients’ baseline and laboratory test data, with multivariate logistic regression further conducted to investigate the independent risk factors for liver cancer progression after hepatectomy.Among the 288 subjects, 159 had adverse outcomes (death or cancer recurrence). Hepatitis B and high levels of aspartate aminotransferase, gamma-glutamyltransferase, alkaline phosphatase (ALP), direct bilirubin, and total bilirubin as well as low level of lymphocyte (LYM) were found to be associated with disease progression in the univariate analysis, and were introduced into the multivariate logistic regression. The results indicated that patients with high ALP level (odds ratio [OR] = 1.004, 95%CI: 1.002–1.007, P = .003) and with a history of hepatitis B (OR = 2.182, 95%CI: 1.165–4.086, P = .015) had a higher risk of liver cancer progression compared with those of lower ALP level and those without hepatitis B respectively, whereas the elevated level of LYM (OR = 0.710, 95%CI: 0.516–0.978, P = .034) had favorable progression.The elevated ALP level and a history of hepatitis B may increase the risk of death or cancer recurrence, whereas high LYM level may decrease poor progression among liver cancer patients after hepatectomy. More importance should be attached to the improvement of the liver function and treatment of hepatitis B to enable a better outcome for the patients.     

Mean platelet volume at baseline and immune thrombocytopenia relapse in Chinese newly-diagnosed patients: a retrospective cohort study

ABSTRACT

Objective: In China, the ability of the current immune thrombocytopenia (ITP) guideline to stratify recurrent risk at diagnosis is limited. This study aimed to investigate whether mean platelet volume at diagnosis (MPV) is a risk factor for ITP relapse in Chinese.

Methods: The present study was a retrospective cohort study. Two hundred thirty-three adult patients with newly diagnosed ITP were consecutively and nonselectively collected from March 2013 to June 2017. The exposure and outcome variable were MPV at baseline and relapse-free survival at 6 months. Other covariants included demographic data, general information, variables that can affect MPV reported by previous literature and risk factors of ITP relapse.

Results: After adjusting potential confounders, the non-linear relationship was detected between MPV and ITP relapse, and inflection point was 21. The effect sizes and the confidence intervals on the left and right sides of inflection point were 1.30 (1.22–1.39) and 0.89 (0.76–1.04), respectively. Subgroup analysis showed, in subjects with hyperuricemia (1.54 (1.24, 1.90)), MPV showed significant differences from non-hyperuricemia (1.19 (1.13, 1.25)), and the p for interaction was less than 0.05.

Conclusion: The relationship between MPV and ITP relapse is non-linear. MPV is an independent risk factor of ITP relapse when MPV is less than 21?fl.     

Novel loci involved in platelet function and platelet count identified by a genome-wide study performed in children

Background

Genome-wide association studies are currently identifying new loci with potential roles in thrombosis and hemostasis: these loci include novel polymorphisms associated with platelet function traits and count. However, no genome-wide study performed on children has been reported to date, in spite of the potential that these subjects have in genetic studies, when compared to adults, given the minimal degree of confounders, i.e., acquired and environmental factors, such as smoking, physical activity, diet, and drug or hormone intake, which are particularly important in platelet function.

Design and Methods

To identify new genetic variants involved in platelet reactivity and count, we performed a genome-wide association study on 75 children (8.5±1.8 years) using the Illumina Sentrix Human CNV370-Quad BeadChip containing 320,610 single nucleotide polymorphisms. Functional analyses included assessment of platelet aggregation and granule secretion triggered by different agonists (arachidonic acid, collagen, epinephrine, ADP), as well as platelet count. Associations were selected based on statistical significance and physiological relevance for a subsequent replication study in a similar sample of 286 children.

Results

We confirmed previously established associations with plasma levels of factors XII, VII and VIII as well as associations with platelet responses to ADP. Additionally, we identified 82 associations with platelet reactivity and count with a P value less than 10⁻⁵. From the associations selected for further replication, we validated two single nucleotide polymorphisms with mildly increased platelet reactivity (rs4366150 and rs1787566) on the LPAR1 and MYO5B genes, encoding lisophosphatidic acid receptor-1 and myosin VB, respectively; and rs1937970, located on the NRG3 gene coding neuroregulin-3, associated with platelet count.

Conclusions

Our genome-wide association study performed in children, followed by a validation analysis, led us to the identification of new genes potentially relevant in platelet function and biogenesis.     

Platelet count evaluation using three automated haematology analysers compared with the immunoplatelet reference method, and estimation of possible inadequate platelet transfusion

The accuracy of three automated haematology analysers [Sysmex XE-2100 (both optical and impedance mode), Bayer Advia 120, and Beckman Coulter LH-750] was compared with the immunoplatelet reference method for platelet measurement. A total of 165 blood specimens were obtained from patients and platelet counts were determined using the four-automated haematology analyser methods and the immunoplatelet reference method. The coefficients of determination (R(2)) between the automated haematology analyser methods and the immunoplatelet reference method for the overall platelet range were >0.98. A bias study, however, showed some disagreement. The use of a coincidence correction calculation for the immunoplatelet method did not improve the correlation between the immunoplatelet method and the automated haematology analyser methods. To estimate the possibility of inadequate platelet transfusion, the number of prophylactic platelet transfusion indications determined by the automated haematology analyser platelet counts were compared with the number of transfusion indications according to the platelet counts determined by the immunoplatelet method. An additional 48 blood specimens were included in this analysis. All of the automated haematology analysers showed some disagreement in the transfusion indications when compared with the immunoplatelet method, suggesting the possibility of inadequate platelet transfusion.     

EDTA-dependent pseudothrombocytopenia: a clinical study of 18 patients and a review of the literature

EDTA-dependent pseudothrombocytopenia (EDTA-PTCP) is the phenomenon of a spurious low platelet count due to the appearance of antibodies that cause platelet agglutination in blood anticoagulated with EDTA. We review here the clinical features of 18 patients with EDTA-PTCP treated in our hospital from 1984 to 1987 as well as those of 34 patients reported in the literature. This phenomenon appears more frequently in severely ill patients, in association with autoimmune, neoplastic, atherosclerosis-related, and liver diseases. In the majority of our patients, EDTA-PTCP appeared during hospitalization, indicating that the antibody is an acquired one. Neither splenomegaly nor the presence of autoimmune markers were features of this entity. Unlike true thrombocytopenias, EDTA-PTCP is associated with a normal mean platelet volume. Awareness of this entity is essential since EDTA-PTCP is frequently misdiagnosed and therefore incorrectly treated.     

The impact of post-hepatectomy liver failure on mortality: a population-based study

Abstract

Background: Post-hepatectomy liver failure (PHLF) is considered a main reason for death after major hepatectomy. The reported PHLF-related mortality differs largely and the data mainly originate from single centers.

Aim: A retrospective, population-based register study was designed to evaluate the impact of PHLF on 90-day mortality after hepatectomy.

Method: All patients who underwent liver resection in Sweden between 2005 and 2009 were retrospectively identified using the Swedish Hospital Discharge Registry. 30- and 90-day mortality were identified by linkage to the Registry of Causes of Death. Additional clinical data were obtained from the medical charts in all seven university hospitals in Sweden. PHLF was defined according to Balzan criteria (Bilirubin >50 µg/L and international normalized ratio >1.5) on postoperative day 5.

Results: A total of 2461 liver resections were performed (2194 in university hospitals). 30- and 90-day mortality were 1.3% and 2.5%, respectively. 90-day mortality at university hospitals was 2.1% (n?=?46). In 41% (n?=?19) of these patients, PHLF alone or in combination with multi-organ failure was identified as cause of death. Between the PHLF and non-PHLF group, there was no significant difference regarding age, sex, American Society of Anesthesiologists-classification, or preoperative chemotherapy. Cholangiocarcinoma as indication for surgery, need for vascular reconstruction and an extended resection were significantly overrepresented in the PHLF-group. Between groups, the incidence of 50:50 criteria differed significantly already on postoperative day 3.

Conclusion: Overall mortality is very low after hepatectomy in Sweden. PHLF represents the single most important cause of death even in a population-based setting.     

Blood collection,platelet isolation and measurement of platelet count and size in mice—a practical guide

Inherited or acquired disorders of platelet production and function can result in thrombocytopenia and bleeding. Mouse models have proven useful for investigating the mechanisms that underlie these defects in humans. Precise methods for blood withdrawal, platelet isolation and measurement of platelet parameters are key for the generation of reproducible and conclusive data.

Here, we provide three different protocols for mouse platelet isolation to encourage research knowledge transfer between experienced laboratories, while at the same time enabling less experienced researchers to implement a protocol that best suits their local expertise and equipment. We also address the issue that reported mouse platelet count and size vary considerably in the literature by investigating different factors that influence these important platelet parameters, namely: 1) genetic background and gender, 2) choice of analysis method (hematological analyzer or flow cytometry), 3) dilution of the blood sample and 4) choice of anticoagulant. The herein presented results and considerations may serve as a practical guide for both experienced and new researchers in the platelet field.     

Ratio of mean platelet volume to platelet count is a potential surrogate marker predicting liver cirrhosis

AIM To provide a simple surrogate marker predictive of liver cirrhosis(LC).METHODS Specimens from 302 patients who underwent resection for hepatocellular carcinoma between January 2006 and December 2012 were retrospectively analyzed. Based on pathologic findings, patients were divided into groups based on whether or not they had LC. Parameters associated with hepatic functional reserve were compared in these two groups using MannWhitney U-test for univariate analysis. Factors differing significantly in univariate analyses were entered into multivariate logistic regression analysis.RESULTS There were significant differences between the LC group(n = 100) and non-LC group(n = 202) in prothrombin activity, concentrations of alanine aminotransferase, aspartate aminotransferase, total bilirubin, albumin, cholinesterase, type Ⅳ collagen, hyaluronic acid, indocyanine green retention rate at 15 min, maximal removal rate of technitium-99 m diethylene triamine pentaacetic acid-galactosyl human serum albumin and ratio of mean platelet volume to platelet count(MPV/PLT). Multivariate analysis showed that prothrombin activity, concentrations of alanine aminotransferase, aspartate aminotransferase, total bilirubin and hyaluronic acid, and MPV/PLT ratio were factors independently predictive of LC. The area under the curve value for MPV/PLT was 0.78,with a 0.8 cutoff value having a sensitivity of 65% and a specificity of 78%.CONCLUSION The MPV/PLT ratio, which can be determined simply from the complete blood count, may be a simple surrogate marker predicting LC.