Garlic flavonoids alleviate H2 O2 induced oxidative damage in L02 cells and induced apoptosis in HepG2 cells by Bcl-2/Caspase pathway.

Liver damage is a common liver disorder, which could induce liver cancer. Oral antioxidant is one of the effective treatments to prevent and alleviate liver damage. In this study, three flavonoids namely myricetin, isoquercitrin, and isorhamnetin were isolated and identified from Laba garlic. The isolated compounds were investigated on the protective effects against H2 O2 -induced oxidative damages in hepatic L02 cells and apoptosis inducing mechanism in hepatic cancer cells HepG2 by using MTT assay, flow cytometry and western blotting analysis. Myricetin, isoquercitrin, and isorhamnetin showed proliferation inhibition on HepG2 cells with IC50 value of 44.32 ± 0.213 µM, 49.68 ± 0.192 µM, and 54.32 ± 0.176 µM, respectively. While they showed low toxicity on normal cell lines L02. They could significantly alleviate the oxidative damage towards L02 cells (P < 0.05), via inhibiting the morphological changes in mitochondria and upholding the integrity of mitochondrial structure and function. The fluorescence intensity of L02 cells pre-treated with myricetin, isoquercitrin, and isorhamnetin (100 µM) was 89.23 ± 1.26%, 89.35 ± 1.43% and 88.97 ± 0.79%, respectively. Moreover, the flavonoids could induce apoptosis in HepG2 cells via Bcl-2/Caspase pathways, where it could up-regulate the expression of Bax and down-regulate the expression of Bcl-2, Bcl-xL, pro-Caspase-3, and pro-Caspase-9 proteins in a dose dependent manner. Overall, the results suggested that the flavonoids from Laba garlic might be a promising candidate for the treatment of various liver disorders. PRACTICAL APPLICATION: Flavonoids from Laba garlic showed selective toxicity towards HepG2 cells in comparison to L02 cells via regulating Bcl-2/caspase pathway. Additionally, the isolated flavonoids expressively barred the oxidative damage induced by H2 O2 in L02 cells. These results suggested that the flavonoids from laba garlic could be a promising agent towards the development of functional foods.

Study of the role of CD4+CD45RClowTreg in regulating immune tolerance of rats undergoing liver transplantation / 器官移植

Objective To investigate the role of CD4+CD45RClow regulatory T cell (Treg) in the immune tolerance induction of rats undergoing liver transplantation. Methods Liver transplantation rat models of acute rejection (AR) [Lewis→Brown Norway (BN), AR group] and spontaneous tolerance (BN→Lewis, tolerance group) were established, with 6 rats in each group. Moreover, 3 Lewis rats and 3 BN rats were assigned into the sham operation group (control group). The liver tissues of rats in each group were subject to pathological staining. The expression of T cell subsets and plasmacytoid dendritic cells (pDC) in the peripheral blood, liver graft and spleen of rats was detected in each group. The correlation between pDC and CD4+CD45RClowTreg was analyzed. The expression levels of CD4, CD45RC and CD103 in the liver graft and spleen of rats were quantitatively measured in each group. Results In the AR group, pathological manifestations mainly consisted of inflammatory cell infiltration and structure disorders of transplant liver. Compared with the AR group, the expression levels of CD4+CD25+Treg and CD8+Treg in the peripheral blood were significantly up-regulated in the tolerance group (all P < 0.05). In the peripheral blood, the expression level of CD4+CD25+Treg was positively correlated with that of CD8+Treg (r=0.742, P=0.022). In the AR group, the expression level of CD4+CD45RChighT cell in the peripheral blood was significantly higher than those in the tolerance and control groups (both P < 0.05). Compared with the AR group, the expression level of CD4+CD45RClowTreg in the spleen, and the expression levels of CD8+CD45RClowTreg in the peripheral blood, transplant liver and spleen were significantly up-regulated in the tolerance group (all P < 0.05). Compared with the control and AR groups, the ratio of CD8+CD45RClowTreg/CD8+T in the peripheral blood and the expression levels of pDC in the peripheral blood, transplant liver and spleen were all significantly up-regulated in the tolerance group (all P < 0.05). The expression level of CD4+CD45RClowTreg was positively correlated with the changes of pDC (r=0.506, P=0.016). The expression levels of CD4, CD45RC and CD103 in the transplant liver and spleen of rats were up-regulated in the tolerance group. In the AR group, the expression levels of CD4 and CD45RC were up-regulated, whereas that of CD103 was down-regulated. Conclusions CD4+CD45RClowTreg is a cell subgroup with negative immune regulation, which may construct a regulatory cell network of immune tolerance induction along with CD8+CD45RClowTreg and pDC.

Risk factors and treatment experience of Pneumocystic carinii pneumonia after liver transplantation / 中华器官移植杂志

Objective:To explore the risk factors of Pneumocystis carinii pneumonia (PCP) after orthotopic liver transplantation (OLT), and optimize the treatment strategy. Methods:From May 2015 to March 2019, patients undergoing OLT and suffering from postoperative PCP were selected into PCP group ( n=8). Using the propensity score matching method, controls without postoperative PCP were selected from concurrent OLT patients at a ratio of 1: 4 ( n=32). Clinical data were collected and counted for analyzing the risk factors of PCP post-OLT. Results:During this period, 385 cases of OLT were performed. The incidence of PCP was 2.1% (8/385). PCP group were all males with an average age of (52.63±12.99)(27-69) years. PCP has an average onset time of (19.88±13.22)(9-50) weeks post-OLT. There were benign liver disease ( n=2) and malignant liver tumor ( n=6). All operative approaches were modified camel OLT. Univariate analysis revealed significant differences in rejection, peripheral blood lymphocyte count and percentage of peripheral blood lymphocyte after OLT ( P<0.05) and no significant differences existed in other factors ( P>0.05). Logistic regression analysis indicated that a lower count of peripheral blood lymphocyte post-OLT was an independent risk factor for postoperative PCP. Conclusions:A lower count of peripheral blood lymphocyte post-OLT elevates the risk of PCP. For high-risk patients, prophylaxis with TMP-SMX (trimethoprim-sulfamethoxazole) may effectively lower the incidence of PCP post-OLT.

Research on clinical efficacy of liver transplantation for severe liver disease / 器官移植

@#Objective To explore the clinical efficacy of liver transplantation for severe liver disease. Methods The clinical data of 51 patients who underwent liver transplantation for severe liver disease were retrospectively analyzed. The general intraoperative conditions were observed, including operation duration, warm ischemia time, cold ischemia time, anhepatic phase, bleeding volume, blood transfusion volume, plasma transfusion volume and so on. The changes in indexes such as total bilirubin (TB), prothrombin time activity (PTA), and prothrombin time international normalized ratio (PT-INR) were observed before operation and at 3 d, 1 week and 2 weeks after operation. The postoperative survival and occurrence of complications were analyzed. The indexes that might affect the prognosis of patients with severe liver disease were analyzed by Cox regression analysis. Results For the 51 patients, operation duration, warm ischemia time and cold ischemia time was 8 (7, 9) h, 3 (2, 3) min and 6 (5, 8) h respectively, intraoperative anhepatic phase was 80 (70, 100) min, intraoperative bleeding volume was 1 000 (550, 1 500) mL, and intraoperative blood transfusion volume was 1 200 (200, 1 600) mL. Postoperative TB, PTA, PT-INR and other indexes improved significantly compared to those preoperatively. Among the 51 patients, 10 cases died, with the death causes of multiple organ failure and severe infection(7 cases), renal insufficiency (2 cases), and cardiovascular complications (1 case). Survival rates at 1 month and 1 year post-transplantation for patients with severe liver disease were 82% and 80%, respectively. Cox regression analysis suggested that abnormal preoperative PTA and PT-INR were the risk factors for post-transplantation death in patients with severe liver disease. Conclusions Liver transplantation significantly improves the survival rate for patients with severe liver disease, perioperative infection prevention and treatment as well as multiple organ function management play key roles in improving post-transplantation survival rate in patients with severe liver disease.

Progress of immunosuppressant management, infection prevention and treatment after liver transplantation in severe liver disease / 器官移植

Liver transplantation is an effective treatment of severe liver disease. However, the pathophysiological changes of patients with severe liver disease are complicated, which significantly increase the difficulty of perioperative management of liver transplantation. Therefore, it is of great significance to strengthen postoperative management of the recipients with severe liver disease after liver transplantation. In this article, the pathophysiological characteristics of severe liver disease, the selection of immunosuppressant after liver transplantation, and the prevention and treatment of infection after liver transplantation in patients with severe liver disease were summarized.

Clinical progress of diagnostic and evaluation criteria for liver transplantation in patients with severe liver disease / 器官移植

Liver transplantation, although recognized as the only effective radical treatment for severe liver disease, might be accompanied by high surgical risks, high perioperative mortality and high postoperative complications. Considering the shortage of donor liver and related surgical risks, it is necessary to strictly control the indication of operation and the opportunity of transplantation. Therefore, accurate diagnosis and comprehensive evaluation of the condition of patients with severe liver disease to be treated by liver transplantation is an important part in determining the treatment plan. At present, there are many evaluation criteria for severe liver disease. In addition to the classic ChildTurcotte-Pugh (CTP) score and model for end-stage liver disease (MELD) score, many other evaluation criteria have also been developed. All transplant centers have their own choices and thus there is no uniform diagnostic criterion, with disputes among various criteria, which is exactly what this paper aims to summarize.

Application of Mingdao immune score system for evaluating the immune status of liver transplant recipients / 中华器官移植杂志

Objective:We proposed a Mingdao immune score system(MISS)to evaluate recipient's immune status after liver transplantation.Methods:From January 2017 to June 2019, retrospective analysis was conducted for 89 recipients of liver transplantation. Age/gender-matched 385 healthy controls(HC)were selected. The percentages of 30 lymphocyte subgroups of patients and HC were measured by flow cytometry. The score of each individual was calculated with our proposed MISS method. And drug concentrations and relevant clinical data were collected.Results:The normal MISS value of a healthy person was 0 score according to our criterion. In this study, the value of MISS for HC was distributed in a nearly normal fashion(-0.73±4.02). When the data from patients at different timepoints were compared, the MISS value started with -1.21±7.42 pre-operation, then declined sharply down to -8.95±8.05 at 1 month and jumped to -4.50±7.80 at 3 months. Afterward it stabilized at -4.18±7.83 between 3~12 months post-operation and finally reached -2.00±5.51 at 1 year ( P<0.05). Patients with acute rejection had higher MISS values than those without acute rejection, ( P<0.05). No significant correlation existed between blood drug concentrations and MISS values ( P>0.05). Conclusions:Our proposed MISS method may reflect the whole immune status. It is useful to manage the application of immunosuppressants in conjunctions with blood drug concentrations and liver graft function.

Hypoxia-stressed cardiomyocytes promote early cardiac differentiation of cardiac stem cells through HIF-1/Jagged1/Notch1 signaling

Hypoxia is beneficial for the differentiation of stem cells transplanted for myocardial injury, but mechanisms underlying this benefit remain unsolved. Here, we report the impact of hypoxia-induced Jagged1 expression in cardiomyocytes (CMs) for driving the differentiation of cardiac stem cells (CSCs). Forced hypoxia-inducible factor 1 (HIF-1) expression and physical hypoxia (5% O) treatment could induce Jagged1 expression in neonatal rat CMs. Pharmacological inhibition of HIF-1 by YC-1 attenuated hypoxia-promoted Jagged1 expression in CMs. An ERK inhibitor (PD98059), but not inhibitors of JNK (SP600125), Notch (DAPT), NF-B (PTDC), JAK (AG490), or STAT3 (Stattic) suppressed hypoxia-induced Jagged1 protein expression in CMs. c-Kit CSCs isolated from neonatal rat hearts using a magnetic-activated cell sorting method expressed GATA4, SM22 or vWF, but not Nkx2.5 and cTnI. Moreover, 87.3% of freshly isolated CSCs displayed Notch1 receptor expression. Direct co-culture of CMs with BrdU-labeled CSCs enhanced CSCs differentiation, as evidenced by an increased number of BrdU/Nkx2.5 cells, while intermittent hypoxia for 21 days promoted co-culture-triggered differentiation of CSCs into CM-like cells. Notably, YC-1 and DAPT attenuated hypoxia-induced differentiation. Our results suggest that hypoxia induces Jagged1 expression in CMs primarily through ERK signaling, and facilitates early cardiac lineage differentiation of CSCs in CM/CSC co-cultures HIF-1/Jagged1/Notch signaling.

High expression of Axl promotes clinical progression of nasopharyngeal carcinoma / 中国病理生理杂志

AIM: To explore the expression and significance of receptor tyrosine kinase anexelekto (Axl) in nasopharyngeal carcinoma (NPC).METHODS: Immunohistochemistry was used to detect the Axl protein expression of 78 patients with NPC and 32 patients with nasopharyngeal chronic inflammation (NPI).The correlations between the Axl protein levels and the clinical parameters of NPC patients were analyzed.NPC cells were cultured in vitro, and the expression of Axl in well differentiated CNE1 cells, poorly-differentiated CNE2Z cells and undifferentiated C666-1 cells was detected by immunofluorescence staining.After treatment of the CNE1and C666-1 cells with Axl specific inhibitor TP-0903, CCK-8 assay was used to detect cell viability, flow cytometry was adopted to analyze the cell cycle distribution, qPCR was used to examine the mRNA levels of Axl and proliferating cell nuclear antigen (PCNA), and Western blot was used to examine the protein expression of Axl and p-Axl.RESULTS: Axl protein was localized in the cell membrane and cytoplasm.The rate of high expression of Axl in NPC was significantly higher than that in NPI (P<0.01).High Axl expression showed no correlations with NPC patients'' age, gender and M stage, while positively correlated with the clinical stage, T stage and N stage (P<0.05).Axl protein showed a low level in the CNE1 cells, but showed a high level in CNE2Z and C666-1 cells.TP-0903 inhibited cell viability in concentration and time dependent manners.TP-0903 at 2 nmol/L showed significant inhibitory effects, as evidenced by arresting the cell cycle at G0 phase and reducing Axl activity and PCNA expression.CONCLUSION: High expression of Axl promotes the clinical progress of NPC.TP-0903 significantly inhibits the viability of NPC cells, suggesting that Axl may be a valuable target in the NPC treatment.