Shaojun Ye

Gypenoside attenuates hepatic ischemia/reperfusion injury in mice via anti-oxidative and anti-apoptotic bioactivities

Gynostemma pentaphyllum is a traditional Chinese medicine that has previously been used for the treatment of chronic inflammation, hyperlipidemia and liver disease. Gypenoside (GP), the predominant component of Gynostemma pentaphyllum, exhibits a therapeutic effect on chronic hepatic injury, fibrosis and fatty liver disease via its anti-inflammatory and anti-oxidant activity. However, the effect of GP on ischemia/reperfusion (I/R)-induced hepatic injury has, to the best of our knowledge, not previously been investigated. In the present study, a hepatic I/R-injury model was successfully established using C57BL/6 mice. In the treatment group, 50 mg/kg GP was administered orally 1 h prior to ischemia. Following hepatic I/R, the levels of hepatic lipid peroxidation and serum alanine aminotransferase increased, while the ratio of hepatic glutathione (GSH):oxidized GSH was reduced, which was effectively attenuated by pretreatment with GP. Furthermore, an increased protein expression of heme oxygenase-1 in the liver tissues of the I/R mice was attenuated by the administration of GP. In addition, the present study indicated that treatment with GP suppressed the I/R-induced increase in the pro-apoptotic protein levels of Bax and cytochrome c and the activity of caspase-3/8, as well as the I/R-induced decrease in the levels of anti-apoptotic protein Bcl-2. In conclusion, the present study indicated that GP effectively protected against I/R-induced hepatic injury via its anti-oxidative and anti-apoptotic bioactivity.

Decreased expression of mitochondrial aldehyde dehydrogenase-2 induces liver injury via activation of the mitogen-activated protein kinase pathway.

The aim of this study was to determine the role of ALDH2 in the injury of liver from brain‐dead donors. Using brain‐dead rabbit model and hypoxia model, levels of ALDH2 and apoptosis in tissues and cell lines were determined by Western blot, flow cytometry (FCM), and transferase (TdT)‐mediated biotin‐16‐dUTP nick‐end labeling (TUNEL) assays. After the expression of ALDH2 during hypoxia had been inhibited or activated, the accumulations of 4‐hydroxynonenal (4‐HNE) and molecules involved in mitogen‐activated protein kinase (MAPK) signaling pathway were analyzed using ELISA kit and Western blot. The low expression of phosphorylated ALDH2 in liver was time‐dependent in the brain‐dead rabbit model. Immunohistochemistry showed ALDH2 was primarily located in endothelial, and the rates of cell apoptosis in the donation after brain‐death (DBD) rabbit groups significantly increased with time. Following the treatment of inhibitor of ALDH2, daidzein, in combination with hypoxia for 8 h, the apoptosis rate and the levels of 4‐HNE, P‐JNK, and cleaved caspase‐3 significantly increased in contrast to that in hypoxic HUVECs; however, they all decreased after treatment with Alda‐1 and hypoxia compared with that in hypoxic HUVECs (P < 0.05). Instead, the levels of P‐P38, P‐ERK, P‐JNK, and cleaved caspase‐3 decreased and the ratio of bcl‐2/bax increased with ad‐ALDH2 (106pfu/ml) in combination with hypoxia for 8 h, which significantly alleviated in contrast to that in hypoxic HUVECs. We found low expression of ALDH2 and high rates of apoptosis in the livers of brain‐dead donor rabbits. Furthermore, decreased ALDH2 led to apoptosis in HUVECs through MAPK pathway.

Mechanisms of Hypothermic Machine Perfusion to Decrease Donation After Cardiac Death Graft Inflammation: Through the Pathway of Upregulating Expression of KLF2 and Inhibiting TGF-β Signaling

Hypothermic machine perfusion (HMP) has been known as an efficient way to improve kidney graft function, but the underlying mechanisms remain unclear. Here, we adopt a rabbit reperfusion mode to investigate the upstream mechanisms of end-ischemic HMP of kidneys from donors after cardiac death (DCD), with static cold storage (CS) as a control. Eighteen New Zealand healthy male rabbits (12 weeks old, with a weight of 3.0 ± 0.2 kg) were randomly divided into three groups: HMP group, CS group, and Normal group (n = 6). The left kidney of rabbits underwent warm ischemia for 25 min through clamping the left renal pedicle and then reperfusion for 1 h. Then the left kidneys were preserved by CS or HMP (4°C for 4 h) ex vivo respectively, after they were autotransplanted and rabbits were submitted to a right nephrectomy. Twenty-four hours after reperfusion, all left renal specimens were collected. Finally, the expression of Krüppel-like factor 2 (KLF2), transforming growth factor-β (TGF-β) and SMAD4 protein in renal cortical tissue were detected by immunoblotting, and the TGF-β and SMAD4 expressions were further confirmed by immunohistochemistry analysis. We found that expression of KLF2 in HMP group was significantly higher than CS group (P = 0.011), while expression of TGF-β and SMAD4 in HMP group were significantly lower than CS group (P = 0.002, P = 0.01, respectively); Compared with normal group, the expression of TGF-β and SMAD4 in HMP and CS group significantly increased (P<0.05). Compared with CS group, TGF-β and SMAD4 protein were equally down-regulated in glomerular and the tubular epithelial cells in HMP group confirmed by immunohistochemistry. In conclusion, HMP may decrease DCD kidneys inflammation through the pathway of upregulating expression of KLF2 and inhibiting TGF-β signaling after transplantation.

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.

Outcome Improvement for Hypothermic Machine Perfusion Versus Cold Storage for Kidneys From Cardiac Death Donors

Organ shortage has led to an increased use of kidneys from cardiac death donors (DCDs), but controversies about the methods of organ preservation still exist. This study aims to compare the effect of machine perfusion (MP) and cold storage (CS) in protecting kidneys harvested from DCDs. 141 kidney pairs from DCDs between July 2010 and July 2015 were included in this randomized controlled study. One kidney from each donor was randomly assigned to MP and the contralateral kidney was assigned to CS. Delayed graft function (DGF) rate, resistance index of renal arteries, early renal function, and survival rates were used to estimate the effect of preservation. The results showed that MP decreased the rate of DGF from 33.3 to 22.0% (P = 0.033). Ultrasound of the kidneys within 48 h after transplantation showed that the resistance index of renal main artery (0.673 ± 0.063 vs. 0.793 ± 0.124, P < 0.001), sub segmental artery (0.66 ± 0.062 vs. 0.764 ± 0.077, P < 0.001) and interlobular artery (0.648 ± 0.056 vs. 0.745 ± 0.111, P = 0.023) were all significantly lower in the MP group than those in the CS group. Furthermore, compared to the CS group, in the first 7 days following transplantation, the median urine volume was significantly higher (4080 mL vs. 3000 mL, P = 0.047) in kidneys sustained using MP and the median serum creatinine was remarkably lower (180 µmol/L vs. 390 µmol/L, P = 0.024). More importantly, MP group had higher 1- and 3-year graft survival rates (98% vs. 93%, P = 0.026; 93% vs. 82%, P = 0.036, respectively). Hypothermic MP improved the outcomes of DCD kidney transplantation.

Association between the overexpression of PBOV1 and the prognosis of patients with hepatocellular carcinoma

Prostate and breast cancer overexpressed 1 (PBOV1) is a known human protein-coding gene with an uncharacterized function; it has been demonstrated to be overexpressed in a variety of human cancer types. The overexpression of PBOV1 has been indicated as significantly associated with the poor prognosis of these types of cancer. However, the function of PBOV1 in hepatocellular carcinoma (HCC) has not yet been elucidated. The present study was designed to evaluate the expression and prognostic significance of PBOV1 in HCC. Reverse transcription-quantitative polymerase chain reaction and western blot analyses were conducted to measure the expression of PBOV1 in HCC cell lines and tissues. The associations between PBOV1 expression and clinicopathological features were statistically analyzed. The association between PBOV1 expression and the prognosis of HCC patients was analyzed by the Kaplan-Meier method. The mRNA and protein expression levels of PBOV1 were significantly increased in the HCC cell lines and HCC tissues (all P<0.05) compared with normal cell lines and tissues. In addition, PBOV1 expression was significantly associated with maximal tumor size (P=0.032), tumor metastasis (P=0.035) and tumor stage (P=0.017). The Kaplan-Meier survival curves indicated that overall survival was significantly poorer in patients with HCC with PBOV1 overexpression (P<0.05) compared with patients with low expression levels. The multivariate analysis indicated that high PBOV1 expression was an independent predictor of poor overall survival. To the best of our knowledge, the data of the present study describes the expression pattern of PBOV1 in HCC for the first time, and also suggests that PBOV1 may serve as a valuable prognostic biomarker for HCC.

Simvastatin ameliorates total liver ischemia/reperfusion injury via KLF2-mediated mechanism in rats

OBJECTIVE The total hepatic ischemia/reperfusion injury (IRI) involves the fact that both liver and gut are subjected to warm ischemia, which is a complex unavoidable process encountered during liver transplantation and a serious threat to graft outcome. The ways to improve hepatic IRI are currently limited. The aim of the present study was to explore the protective effect of simvastatin on total hepatic IRI and examine the underlying mechanisms. METHODS Male Sprague Dawley rats were subjected to total (100%) hepatic warm ischemia to induce hepatic IRI. Thirty-six male rats (250-300 g) were randomly divided into three groups: sham, IRI control and simvastatin (1 mg/kg) pretreatment 0.5 h before surgery. Serum samples and liver tissues were collected after reperfusion at 6 and 24 h for further studies. RESULTS Simvastatin pretreatment significantly decreased the values of the transaminases alanine aminotransferase and aspartate aminotransferase and improved histological alterations according to improved Suzukis Score (P < 0.05). Moreover, simvastatin upregulated the expression of Kruppel-like factor 2 (KLF2), phosphorylated endothelial nitric oxide synthase and thrombomodulin (P < 0.05). Furthermore, simvastatin pretreatment affected superoxide dismutase and malondialdehyde activities (P < 0.05) to reduce oxidative stress, and inhibited levels of high-mobility group box-1, CD68, toll-like receptor 4, tumor necrosis factor α, interleukin-1β and interleukin-6 (P < 0.05) to suppress inflammatory response. CONCLUSION Simvastatin pretreatment ameliorates total hepatic IRI via a KLF2-mediated protective mechanism. Simvastatin may be used as a potential prophylactic treatment strategy for clinical trials against hepatic IRI.

Long-Term Outcomes of Ante-Situm Resection and Auto-Transplantation in Conventionally Unresectable Hepatocellular Carcinoma: A Single-Center Experience

Background Ante-situm resection and auto-transplantation (ante-situm for short) provides a more aggressive approach to conventionally unresectable hepatocellular carcinoma (HCC). We described the long-term outcomes of patients with HCCs who underwent this technique. Material/Methods Between October 2005 and December 2016, we performed 23 ante-situm liver resections. We evaluated postoperative complications, 90-day mortality, recurrence, and long-term survival rates, and reviewed the literature on this topic. Results Five types of complications associated with six patients were observed.: 1) primary nonfunctioning liver, thus receiving a liver transplantation; 2) initial poor liver function with recovery two weeks after treatment; 3) diagnoses of portal vein tumor thrombosis, biliary fistula, and small-for-size syndrome, respectively. The median follow-up was 3.6 years; 12 out of 23 patients were alive at the end of the study. One patient who had hepatic recurrence was lost to follow-up after three months. One patient died of multiple organ dysfunction syndrome after the operation, nine patients died due to hepatic recurrence and/or extrahepatic metastasis of HCC. The one-year, three-year, five-year, and 10-year survival rates were 65.2%, 56.5%, 50.9%, and 20.3%, respectively. The one-year, three-year, five-year, and 10-year recurrence rates were 60.9%, 50.7%, 50.7%, and 50.7%, respectively. The chi-square test revealed the patients with recurrence after ante-situm technique were more likely to have poor prognosis (mortality of patients with recurrence versus no-recurrence: 88.9% versus 14.3%, p<0.05) and a strong association was evidenced by Cramer’s V statistic (Cramer’s V=0.734). Conclusions Ante-situm procedure showed benefits in select patients with HCCs who had contraindications for conventional resection operations. In our case series, the ante-situm technique resulted in lower mortality compared to other ex-vivo hepatic resection techniques reported in the literature and similar long-term efficacy compared to cases of HCCs suitable for conventional resections. HCCs recurrence was a major risk factor associated with the survival rate of ante-situm technique.

Polyaniline promotes peripheral nerve regeneration by enhancement of the brain‑derived neurotrophic factor and ciliary neurotrophic factor expression and activation of the ERK1/2/MAPK signaling pathway

A previous study has demonstrated a progression in the nerve regeneration by polyaniline/cellulose (PANI/RC), although the underlying mechanism was not elucidated. In the present study, regenerated nerves were investigated, using histological techniques, functional assays and western blot analysis. The triceps surae muscle weight ratio percentages of the sham, regenerated cellulose (RC) and the PANI/RC groups were 38.88±4.76 and 76.32±7.11%, respectively. The thickness of the myelin sheath for the aforementioned groups were as follows: 1.2±0.27; 0.49±0.21 and 0.93±0.28 µl. Western blot analysis demonstrated that the ciliary neurotrophic factor (CNTF) and brain-derived neurotrophic factor (BDNF) were highly expressed in the regenerated nerve in the presence of polyaniline. Phosphorylated extracellular kinase (p-ERK)1/2 expression in the PANI/RC group was significantly elevated compared with the RC group (1.83-fold) and the sham group (4.92-fold). The expression of the axon sprout-associated proteins, such as Tau, α-tubulin and growth associated protein-43, were increased (1.64, 1.59 and 1.24-fold, respectively) compared with the RC group. The results demonstrated that PANI enhances the expression and secretion of BDNF and CNTF, activates the ERK1/2 signaling pathway and increases the expression levels of the GAP-43, Tau and α-tubulin, suggesting an insight into nerve regeneration and possible clinical interventions in nerve injury.