Successful rescue of renal transplantation with cardiac arrest after electrical storm: A case report.

RATIONALE: Most patients with end-stage chronic kidney disease are associated with complications such as renal hypertension, renal anemia, hyperkalemia, water-sodium retention, and disorders of acid-base balance after long-term renal replacement therapy, which can lead to increased cardiac burden, some degree of myocardial damage, and finally progress to arrhythmia and heart failure. These are the main reasons why patients with chronic kidney disease are prone to cardiovascular events after renal transplantation. PATIENT CONCERNS: We report a case of sudden onset of ventricular fibrillation on the postoperative second day, with repeated electrical storm accompanied by cardiac arrest during resuscitation, a very long cardiopulmonary resuscitation (CPR) process of 5 hours and 14 minutes, and >20 cycles of cardiac defibrillation. DIAGNOSES: According to the patient history and resuscitation process, a diagnosis of ES with cardiac arrest after renal transplantation was formulated. INTERVENTION: According to the American Heart Association guidelines for CPR and cardiovascular emergencies, resuscitation measures such as CPR, tracheal intubation, electric defibrillation, symptomatic medication, etc. were performed on the patient. OUTCOMES: Finally, the patient was successfully resuscitated, after which the patient had stable respiratory circulation and no neurological complications. To our knowledge, this is the only reported case in which a patient survived with good neurologic outcomes after a resuscitation that lasted as long as 5 hours and 14 minutes. LESSONS: This case of adequate resuscitation can provide experience and a basis for CPR of patients with in-hospital complications of cardiovascular events for a long time.

Construction of a ceRNA network and a genomic-clinicopathologic nomogram to predict survival for HBV-related HCC.

Some lncRNA-associated competing endogenous RNAs (ceRNAs) are considered as potential biomarkers for targeted therapies and prognosis in human cancer. In our present study, we aimed to construct a ceRNA network and establish a genomic-clinicopathologic nomogram to provide insights into the molecular mechanisms and predict survival for HBV-related HCC. The Cancer Genome Atlas (TCGA) database was applied to collect the data of LIHC RNA-seq dataset and miRNA-seq dataset as well as the clinicopathological information. Identification of differentially expressed RNAs (mRNAs, lncRNAs, and miRNAs) between HBV-related HCC samples and normal samples was conducted using Limma package in R. The Database for Annotation, Visualization, and Integrated Discovery (DAVID) was used for performing the functional enrichment analysis of differentially expressed mRNAs. The ceRNA network was carried out using Cytoscape. The LASSO-penalized Cox regression analysis was implemented to identify HCC-related lncRNAs, and the multivariate Cox regression analysis was conducted for the establishment of a genomic-clinicopathology nomogram. A total of 1859 DEmRNAs, 113 DElncRNAs, and 89 DEmiRNAs were screened out etween HBV-related HCC samples and normal samples. A ceRNA network including 44 DEmRNAs, 7 DElncRNAs, and 20 DEmiRNAs was constructed. 7 DElncRNAs (PVT1, LINC01138, LINC02499, AL355488.2, FGF14-AS2, MAFG-AS1 and LINC00261) were finally identified as prognostic indicators. The area under the curve reached 0.8169 for the 7-lncRNA signature. The predictive accuracy and clinical application value were remarkably high for the genomic-clinicopathologic nomogram integrating the histological grade and the 7-gene-based prognostic index. Taken together, we have established a ceRNA network with HBV-related HCC-specific DElncRNAs, DEmiRNAs, and DEmRNAs. Furthermore, the genome-wide data of lncRNA expression were analyzed using the TCGA database, and a 7-lncRNA signature was identified as a potential prognostic predictor for HBV-related HCC patients. Novel functional studies were provided by our current findings for elucidating the molecular mechanism of lncRNA in HBV-related HCC.

Double-lobectomy in a steatotic liver transplantation rat model.

Establishing a steatotic liver transplantation animal model can be a challenging process, which requires complex microsurgical technologies. The present study established a novel rat model of stable steatotic liver transplantation for marginal liver graft research, which notably minimized the number of animals used for the experiment. Briefly, male Sprague-Dawley rats (n=90) were fed with a high-fat diet (HFD; 60%, kJ) or standard chow diet (SCD) for 8 weeks. The liver enzymes and lipid levels were assessed every week, and the degree of steatosis was determined via hematoxylin and eosin and Oil Red O staining. The results demonstrated that there were no significant differences in alanine aminotransaminase and aspartate aminotransferase levels between the SCD and HFD groups (P>0.05), whereas the level of plasma triglyceride (TG) increased by 1.76-fold in the HFD group at week 2, and progressively decreased to baseline levels by week 8. Significantly higher levels of TG were observed in the HFD group compared with the SCD group at week 2 (P<0.05). In addition, the levels of plasma glucose (P<0.05), portal insulin (P<0.05) and content of liver lipid (P<0.01) increased in the HFD group compared with the SCD group. After 6 weeks, the liver steatosis was successfully formed and stable. Consequently, a rat liver developed hepatic macrovesicular steatosis >60%, which was subsequently used for transplantation after double-lobectomy. Post-transplantation survival rates in the HFD and SCD groups were as follows: Week 1, 80 vs. 100% and 1 month, 20 vs. 100%. A total of 20 rats were not sacrificed by performing double-lobectomy for biopsy. Taken together, the results of the present study suggest that rat liver double-lobectomy may be safely applied in steatotic liver transplantation without the need to sacrifice a large number of animals.

Bioinformatics analysis of LINC01554 and its co­expressed genes in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a leading cause of cancer­related morbidity and mortality globally. Despite the remarkable improvements in comprehensive HCC treatment, the underlying mechanistic details of HCC remain elusive. We screened HCC patients for differentially expressed genes (DEGs) using the Gene Expression Omnibus (GSE113850) and The Cancer Genome Atlas (TCGA) datasets. LINC01554 expression in 40 paired samples was determined by quantitative reverse transcription polymerase chain reaction (RT­qPCR), and its clinical significance was assessed. LINC01554 was found to have a gain­of­function role in HCC in vitro. Additionally, the bioinformatics analysis of the genes co­expressed with LINC01554 was performed using the Co­LncRNA website, and potential molecular mechanisms were investigated using the Gene Ontology and Kyoto Encyclopaedia of Genes and Genomes resources and validated by in vitro experiments. A total of 229 DEGs were identified from the GSE113850 dataset. Among the identified DEGs, three long non­coding RNAs (lncRNAs) (DIO3OS, LINC01554, and LINC01093) with |logFC| ≥2 and P<0.05 were screened. A total of 148 lncRNAs with |logFC| ≥1 and P<0.05 were identified from TCGA dataset. Low LINC01554 expression levels were significantly correlated with overall survival, pathological stage, hepatitis B infection, tumour size, portal vein tumour thrombus, and TNM stage. Using gain­of­function assays, we further showed that LINC01554 inhibited the proliferation, migration, and invasion of the HCCLM9 and SK­Hep1 cells and promoted G0/G1 arrest, but it did not significantly affect apoptosis. Western blotting revealed that LINC01554 overexpression resulted in increased ZO­1 and E­cadherin expression levels, but decreased N­cadherin and vimentin expression levels. Moreover, LINC01554 overexpression inhibited Akt, p­Akt, ß­catenin, and p­Gsk3ß expression. Our results showed that LINC01554 repressed HCC cell invasiveness and epithelial­to­mesenchymal transition partly by inhibiting Wnt and PI3K­Akt signalling in vitro. Taken together, our findings provide new insights into the molecular mechanisms underlying HCC tumourigenesis and implicate LINC01554 as a potential target for HCC therapy.

LINC00844 promotes proliferation and migration of hepatocellular carcinoma by regulating NDRG1 expression.

BACKGROUND: Aberrant expression of long noncoding RNAs are implicated in the pathogenesis of human malignancies. LINC00844 expression is dramatically downregulated in prostate cancer, and functional studies have revealed the association between the aberrant expression of LINC00844 and prostate cancer cell invasion and metastasis. However, the function and mechanism of action of LINC00844 in the pathogenesis of hepatocellular carcinoma (HCC) are poorly understood. METHODS: LINC00844 and N-Myc downstream-regulated 1 (NDRG1) expression in HCC tissues and cell lines was detected with real-time quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. Correlations between LINC00844 expression level and clinicopathological features were investigated using the original data from The Cancer Genome Atlas (TCGA) database. HepG2 and HCCLM9 cell lines were transfected with Lv-LIN00844 virus to obtain LINC00844-overexpressing cell lines. Cell proliferation and cell invasion and migration were examined with the cell counting kit-8 (CCK-8) and transwell assay, respectively. Furthermore, the correlation between LINC00844 and NDRG1 expression was analysed using Pearson's correlation analysis. RESULTS: LINC00844 expression was significantly downregulatedin HCC tissues and cell lines, and a statistical correlation was detected between low LINC00844 expression and sex (Female), advanced American Joint Committee on Cancer (AJCC) stage (III + IV), histological grade (G3 + G4), and vascular invasion (Micro and Macro). In vitro experiments showed that LINC00844 overexpression significantly repressed the proliferation, migration, and invasion of HCC cells. NDRG1 expression was higher in HCC tissues and LINC00844 could partly inhibit the expression of NDRG1.

Cytochrome b5 Interacts With Cytochrome C and Inhibits Hepatocyte Apoptosis in Brain-dead Rabbit Donors.

BACKGROUND: Donation after brain death (BD) liver grafts undergo the process of hypoxia-ischemia, which induces hepatocyte apoptosis, but the underlying mechanisms remain unclear. Cytochrome (Cyt) b5 expression was shown to be low in BD rabbits. This study aimed to investigate if Cyt b5 and Cyt c are involved in liver apoptosis after BD. METHODS AND RESULTS: Liver tissue samples were obtained from donors after BD and from BD rabbit models. Tissues were analyzed by immunofluorescence, western blotting, and reverse-transcriptase polymerase chain reaction to detect Cyt b5 and Cyt c protein expressions and mRNA. Normal liver cells (LO-2) were cultured under serum deprivation and hypoxia, and analyzed as above. Cyt b5 protein and mRNA levels had decreased, while Cyt c levels had increased in BD liver donors and rabbits. Similar results were obtained in LO-2 cells cultured under hypoxia. After 6 and 12 hours of serum deprivation and hypoxia, apoptosis was increased, the levels of Cyt b5 gradually decreased, and the levels of Cyt c gradually increased over time; meanwhile, the Cyt b5-Cyt c combination was gradually reduced. A negative linear correlation between Cyt b5 and Cyt c was also observed. CONCLUSIONS: Cyt b5 might be an anti-apoptotic protein that could protect the liver after BD and this protective effect might involve increased binding to Cyt c. This study provides some clues for improving the quality of donor livers.

Arsenic trioxide is an immune adjuvant in liver cancer treatment.

Tumor cells are inhibited effectively by As2O3in vitro and in vivo, although the underlying immune regulatory mechanisms remain unknown. Regulatory T cells play a key role in tumor immune escape. In the present study, we aimed to assess the in vivo effects of As2O3 on the immune status in hepatic cancer and its in vitro regulatory role in cytokine-induced killers(CIKs)cytotoxicity. In a tumor H22 xenograft model of hepatic cancer, we demonstrated that As2O3 treatment decreased tumor volumes and weights, and improved survival by reducing Tregs infiltration into the tumor. Moreover, our data indicated that the exact immune regulatory mechanism of As2O3 might involve elevated CD3+T lymphocyte amounts more than reduced Tregs levels. Furthermore, As2O3 significantly improved CIKs cytotoxicity in vitro by decreasing CD4+T lymphocytes and Tregs, and increasing CD8+T lymphocytes. Our results suggested that As2O3 might act as an immune adjuvant in liver carcinoma treatment by increasing T lymphocytes and decreasing Treg infiltrated into the tumor.

[Acquirement of autologous murine cytotoxic T lymphocytes via cryopreservation of lymphocytes].

Objective To evaluate the effects of cryopreservation on the proliferation and killing activity of lymphocytes, and explore a novel protocol of preparing autologous mouse cytotoxic T lymphocytes (CTLs). Methods Mononuclear cells derived from spleen (5.0×106/mL) were cryopreserved in CELLBANKER2 for 6 days and recovered in water bath at 39DegreesCelsius. The fresh lymphocytes and post-cryopreservation lymphocytes were induced by CD3 mAb (100 ng/mL) and recombinant mouse interleukin 2 (rmIL-2, 100 ng/mL) to obtain cytokine-induced killer cells (CIKs). Dendritic cells (DCs) were co-cultured with fresh allogenic lymphocytes and post-cryopreservation autologous lymphocytes to obtain CTLs. The viable cells were counted by trypan blue staining; the percentages of CD3+T cells and regulatory T cells (Tregs) were determined by flow cytometry; the levels of supernatant IFN-γ were detected through ELISA and the cytotoxicity was evaluated by lactate dehydrogenase (LDH) assay. Results The rate of viable lymphocytes declined to 78% after cryopreservation, and there were no significant differences in the percentages of CD3+T cells and Tregs between pre-cryopreservation and post-cryopreservation. There were no significant differences in the proliferation of Tregs, the level of IFN-γ and the cytotoxicity between the fresh CIKs and cryopreservation CIKs, and the similar results were get between the autologous CTLs and allogenic CTLs. Conclusion The autologous CTLs acquired via cryopreservation of lymphocytes is equivalent to the allogenic CTLs with the similar proliferation and killing activity in vitro.

The organ preservation and enhancement of donation success ratio effect of extracorporeal membrane oxygenation in circulatory unstable brain death donor.

Between 2010 and 2013, we recorded 66 cases of failed organ donation after brain death (DBD) due to the excessive use of the vasoactive drugs resulting in impaired hepatic and/or renal function. To investigate the effect of extracorporeal membrane oxygenation (ECMO) in donor management, ECMO was used to provide support for DBD donors with circulatory and/or respiratory failure from 2013 to 2015. A retrospective cohort study between circulatory non-stable DBD with vasoactive drugs (DBD-drug) and circulatory non-stable DBD with ECMO (DBD-ECMO) was designed to compare the transplant outcomes. A total of 19 brain death donors were supported by ECMO. The incidence rate of post-transplant liver primary non-function (PNF) was 10% (two of 20) in DBD-drug group and zero in DBD-ECMO group. Kidney function indicators, including creatinine clearance and urine production, were significantly better in DBD-ECMO group, as well as the kidney delayed graft function (DGF) rate was found to be decreased by the use of ECMO in our study. Donation success rate increased steadily from 47.8% in 2011 to 84.6% in 2014 after the ECMO intervention. The use of ECMO in assisting circulatory and respiratory function of DBD can reduce liver and kidney injury from vasoactive drugs, thereby improving organ quality and reducing the organ discard rates.

Hypothermic machine perfusion increases A20 expression which protects renal cells against ischemia/reperfusion injury by suppressing inflammation, apoptosis and necroptosis.

There is an urgent need to improve the quality of donor organs obtained after cardiac death. In the present study, we examined the potential mechanisms through which A20 protects renal cells against ischemia/reperfusion injury (IRI) following either hypothermic machine perfusion (HMP) or static cold storage (CS) of the kidneys in a rabbit model. The expression of markers of apoptosis, necroptosis and inflammation in frozen kidney tissues were detected by western blot analysis, RT-qPCR and ELISA. Compared with the CS group, A20 expression was significantly higher in the tissue from the HMP group (P<0.01). By contrast, the expression of nuclear factor-κB (NF-κB) and tumor necrosis factor-α (TNF-α) was significantly lower in HMP group (P<0.01), whereas IκBα expression was significantly higher (P<0.01). The expression of apoptosis signal-regulating kinase 1 (ASK1), phosphorylated (p-)c-Jun N-terminal kinase (JNK) and activated caspase-3 in the HMP group was significantly downregulated compared with that in the CS group (all P<0.01). In addition, A20 inhibited receptor-interacting protein kinase 3 (RIPK3)­mediated necroptosis in the kidney. RIPK3 expression in the HMP group was significantly lower than that in the CS group (P<0.01), although the levels in both groups were higher than those in the sham group (P<0.01). Based on these findings, we propose a novel mechanism underlying the anti-apoptotic effect of A20 in renal cells in which A20 binds to ASK1 and promotes the degradation of ASK1 leading to the suppression of JNK activation and eventually, to the blockade of apoptosis. Thus, HMP reduces inflammation, apoptosis and necroptosis by upregulating the expression of A20; this mechanism may be responsible for protecting the kidney against IRI.

Knockdown by shRNA identifies SLC44A5 as a potential therapeutic target in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has been ranked the second leading cause of cancer­associated mortality in China and the third leading cause of cancer­associated mortality worldwide. A number of previous studies investigating SLC44A5 have revealed important biological insight and disease­specific functions. Therefore, the present study investigated the expression of SLC44A5 in HCC tissues and cell lines, and assessed the effect of SLC44A5 on the viability, cell cycle, apoptosis and invasion of HCC cell lines. The mRNA expression of SLC44A5 in 35 HCC tissues was significantly higher compared with that in 35 normal tissues. The protein expression of SLC44A5 was notably high in MHCC­97H and SMMC­7721 cells compared with that in four other HCC cell lines. Knockdown of SLC44A5 using short hairpin RNA inhibited cell viability and arrested the cells in G1 of the cell cycle by reducing the expression of cell cycle markers, proliferating cell nuclear antigen and cyclin­dependent kinase 2 in MHCC­97H and SMMC­7721 cells. Furthermore, SLC44A5 knockdown cells also exhibited cell apoptosis by reducing the expression levels of apoptosis markers, caspase­3 and caspase­9 in MHCC­97H and SMMC­7721 cells, and suppressed invasion. The present results suggested that SLC44A5 is involved in HCC carcinogenesis and progression in HCC, indicating that SLC44A5 may be a molecular target in cancer therapy.

Peroxiredoxin 6 attenuates ischemia­ and hypoxia­induced liver damage of brain­dead donors.

Oxidative stress induced by ischemia and hypoxia in the livers of donors after brain death (DBD) is associated with poor organ function and low patient survival rates in those receiving DBD liver transplants. Peroxiredoxin 6 (Prdx6) can defend cells against liver damage induced by oxidative stress. The present study aimed to investigate the role of Prdx6 in ischemia­ and hypoxia­induced liver damage in DBD livers. Liver tissue samples from ten DBD patients were collected. The control group constituted of six liver samples from patients with liver hemangioma that had accepted tumor excision surgery. Protein expression levels were determined by western blotting, cell viability was assessed using a CCK­8 assay, intracellular reactive oxygen species (ROS) levels were measured using a ROS assay kit, and phospholipase A2 (PLA2) activity was measured using a PLA2 assay kit. In DBD liver samples, Prdx6 expression was downregulated and the nuclear factor­κB (NF­κB) signaling pathway was activated. Furthermore, when human liver L02 cells were exposed to ischemia and hypoxia, the expression of Prdx6 was reduced, causing an increase in reactive oxygen species (ROS); this in turn activated NF­κB signaling and lowered cell viability (P<0.01). In agreement, overexpression of Prdx6 reduced ROS levels and improved cell viability. It was also demonstrated that inhibition of NF­κB increased Prdx6 expression, while inhibition of Prdx6 limited PLA2 activity, exacerbating ischemia­ and hypoxia­induced cell damage. This data suggests that Prdx6 and its PLA2 activity have a protective role in DBD livers, the expression of which is regulated by NF­κB. Thus, Prdx6 may be a novel target to alleviate liver damage in DBD.