Laparoscopic simultaneous anterograde inguinal and pelvic lymphadenectomy for penile cancer: two planses, three holes, and six steps.

Objective: To assess the feasibility, safety, and efficiency of simultaneous anterograde video laparoscopic inguinal and pelvic lymphadenectomy for penile cancer. Materials and methods: We reviewed retrospectively the records of 22 patients (44 lateral) who underwent inguinal lymph nodes dissection for penile cancer. The procedure was standardized as two planes, three holes, and six steps. Two Separate-planes: superior plane of eternal oblique aponeurosis/ / fascia lata; inferior plane of superficial camper fascia. Three holes: two artificial lateral boundary holes, the internal and external boundary holes, and the hole of oval fossa. Six steps: separate the first separate-plane; separate the second layer; separate two artificial lateral boundary holes; free great saphenous vein; separate the third hole and clean up the deep inguinal lymph nodes; pelvic lymphadenectomy. Results: A total of 22 cases were included and 9 patients underwent simultaneous pelvic lymphadenectomy. The average operation time on both sides was 7.52 ± 3.29 h, which was 0.5-1 h/side after skilled. The average amount of bleeding was 93.18 ± 50.84 ml. A total of 8 patients had postoperative complications, accounting for 36.36%, and no complications great than Clavien-Dindo class III occurred. Conclusion: This study demonstrated that the video laparoscopic simultaneous anterograde inguinal and pelvic lymphadenectomy is a feasible and safe technique. Indocyanine Green was helpful for lymph node identify.

Urine-based regenerative RNA biomarkers for urinary bladder wound healing.

Background: This study aimed to investigate the expression of regeneration-related genes in canine urine during bladder repair. Materials & methods: Canine urine samples were collected after partial cystectomy. Regenerative mRNA of hypoxia-inducible factor (HIF), vascular endothelial growth factor (VEGF), key stem cell transcription factors and cholinergic signals were detected. Results: HIF-1α, VEGF, CD44, IL-6 and prominin-1 expression in canine urine after partial cystectomy exhibited two similar peaks at ∼2 weeks. HIF-1α and VEGF expression were higher in the afternoon than the morning. The expression of key stem cell transcription factors and cholinergic signals also exhibited a rhythm along with bladder healing. Conclusions: The expression of HIF-1α, VEGF, key stem cell transcription factors and cholinergic signals exhibited a time curve distribution during canine bladder healing. The expression trend of some regenerative genes was similar during bladder healing, and a cooperative effect may exist.

Curcumin Attenuates Oxaliplatin-Induced Liver Injury and Oxidative Stress by Activating the Nrf2 Pathway.

PURPOSE: Oxaliplatin (OXA)-induced liver injury is one of the main limiting factors affecting the efficacy of OXA-based chemotherapy in patients with colorectal liver metastases. In addition, oxidative stress is an important pathophysiological mechanism of OXA-induced liver injury. Therefore, dietary antioxidants may decrease or prevent hepatic toxicity in vivo and be beneficial to OXA-based chemotherapy. METHODS: An experimental OXA-induced liver injury animal model was established, and the protective effects of curcumin (CUR) against OXA-induced liver injury were investigated. ELISA was used to determine the levels of MDA, SOD, CAT, and GSH in liver tissue. The effect of CUR treatment on the expression of cytokines and the Nrf2 pathway was determined by real-time PCR and Western blotting. RESULTS: CUR treatment alleviated OXA-induced hepatic pathological damage and splenomegaly. The protective effect of CUR was demonstrated to be correlated with inhibition of oxidative stress, inflammation, and the coagulation system. Furthermore, Western blotting revealed that CUR treatment reverses the suppression of Nrf2 nuclear translocation and increases the expression of HO-1 and NOQ1 in mice with OXA-induced liver injury. Moreover, the Nrf2 activation and hepatoprotective effect of CUR were abolished by brusatol. CONCLUSION: Curcumin attenuates oxaliplatin-induced liver injury and oxidative stress by activating the Nrf2 pathway, which suggests that CUR may be potentially used in the prevention and treatment of OXA-induced liver injury.

Irf5 deficiency in myeloid cells prevents necrotizing enterocolitis by inhibiting M1 macrophage polarization.

Necrotizing enterocolitis (NEC) is a life-threatening inflammatory disease in newborns, but the mechanisms remain unclear. Interferon regulatory factor 5 (IRF5) is a master regulator of macrophage function and is essential for proinflammatory M1 macrophage polarization. Our previous data indicated that M1 macrophages promote NEC injury. Here, we investigated whether IRF5 is involved in the pathogenesis of NEC. First, we found that IRF5 was upregulated in infiltrated macrophages in human neonates with NEC compared to controls. We further confirmed IRF5 upregulation in macrophages in experimental murine NEC and that the infiltrated macrophages were predominantly polarized into the M1 but not the M2 phenotype. Myeloid-specific deficiency of Irf5, which was associated with reduced M1 macrophage polarization and systematic inflammation, dramatically prevented experimental NEC. Moreover, we found that the ablation of Irf5 in myeloid cells markedly suppressed intestinal epithelial cell apoptosis and further prevented intestinal barrier dysfunction in experimental NEC. Bioinformatic and chromatin immunoprecipitation analysis further showed that IRF5 binds to the promoters of the M1 macrophage-associated genes Ccl4, Ccl5, Tnf, and Il12b. Overall, our study provides evidence that IRF5 participates in the pathogenesis of NEC, while the deletion of Irf5 in myeloid cells prevents NEC via inhibiting M1 macrophage polarization.

Oxaliplatin aggravates hepatic oxidative stress, inflammation and fibrosis in a non­alcoholic fatty liver disease mouse model.

Oxaliplatin (OXA)­based chemotherapy is widely used in the treatment of gastrointestinal tumors; however, it is associated with chemotherapy­associated liver injury. Whether OXA induces liver injury and aggravates the already existing hepatic oxidative stress, inflammation and fibrosis in non­alcoholic fatty liver disease (NAFLD), and whether these effects can be alleviated by reduced glutathione (GSH) treatment, remains unclear. In the present study, OXA induced acute liver injury in NAFLD mice. Moreover, OXA increased the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased the levels of superoxide dismutase and GSH peroxidase in the livers of NAFLD mice. OXA also induced the upregulation of hepatic inflammatory cytokines, such as tumor necrosis factor (TNF)­α, interferon (IFN)­Î³ and interleukin (IL)­17, in NAFLD mice. Furthermore, collagen fiber deposition in liver tissues was increased and the expression of transforming growth factor (TGF)­ß, α­smooth muscle actin (SMA) and tissue inhibitor of metallopeptidase (TIMP)­1 was upregulated in the livers of OXA­treated NAFLD mice. Treatment with exogenous GSH alleviated OXA­induced acute liver injury in NAFLD mice, and significantly reduced the levels of ROS, MDA and TNF­α. However, GSH treatment did not inhibit collagen fiber deposition, although it reduced the levels of IFN­Î³, IL­17, TGF­ß, α­SMA and TIMP­1 in the livers of OXA­treated NAFLD mice. In conclusion, OXA chemotherapy may induce acute liver injury and aggravate the existing hepatic oxidative stress, inflammation and fibrosis in NAFLD. Treatment of NAFLD mice with exogenous GSH alleviated OXA­induced liver injury, possibly by ameliorating OXA­aggravated hepatic oxidative stress and inflammation; it did not, however, attenuate OXA­aggravated liver fibrosis.

The hepatoprotective role of reduced glutathione and its underlying mechanism in oxaliplatin-induced acute liver injury.

Currently, the underlying mechanism of oxaliplatin (OXA) induced live injury is unclear. In addition, there is no standard clinical treatment for OXA-induced acute liver injury (ALI). In this study, we established an animal model of OXA-induced ALI, and studied the role of oxidative stress in OXA-induced ALI and the impacts of reduced glutathione (GSH) treatment on OXA-induced ALI. To establish an OXA-induced ALI model, KM mice received intraperitoneal injection of OXA (8 mg/kg) for 4 days. Serum alanine aminotransferase (ALT), aspartate aminotransferase levels (AST), hepatic pathology and oxidative stress indicators in liver tissues were analyzed. To study the impact of GSH treatment on OXA-induced ALI, mice were treated with GSH (400 mg/kg, i.p). In this ALI mouse model, ALT and AST levels were significantly increased (P<0.01). Liver pathological examination revealed varying degrees of liver cell turbidity and degeneration, even balloon-like changes and focal necrosis, and sinusoidal hemorrhage in some cells. Compared with control group, the malondialdehyde (MDA) and GSH levels were significantly increased in OXA-treated group (P<0.01), while the superoxide dismutase SOD and GSH-peroxidase levels were decreased after OXA withdrawal (P<0.01). When GSH was used to treat OXA-induced ALI mice, the pathological injury of liver tissues was alleviated, and serum ALT and AST were significantly decreased. In addition, GSH treatment could reduce the OXA-induced increase of MDA level (P<0.05) in liver tissues, but had no impact on SOD level (P>0.05). We have successfully established an OXA-induced ALI model. Using this model, we discover that oxidative stress plays an important role in OXA-induced ALI. GSH-based hepatoprotective therapy can partially inhibit oxidative stress and alleviate OXA-induced ALI.

Compliance with National Guidelines on the Treatment of Stage II­IVB Nasopharyngeal Carcinoma in a Regional Cancer Center of Southern China

Objective: It is unknown whether the treatment provided to patients with stage II-IVB NPC in southern China adheres to the 2015 NCCN guidelines. Consequently, a retrospective analysis was conducted, in order to evaluate the compliance with NCCN guidelines and identify the areas for improvement. Methods: The present study was a retrospective study that included patients with stage II-IVB NPC in southern China during the period 2013 and 2014. The treatment regimens were compared with the 2015 NCCN guidelines in order to identify potential noncompliance regarding the treatment for stage II­IVB NPC. The statistical analyses included descriptive statistics, univariate and/or multivariate analysis using SPSS version 16.0.0. Results: A total of 215 patients, including 166 men (77.21%) and 49 women (22.79%), were involved in the analysis. Although the overall rate of noncompliance with the NCCN recommendations was 23.26%, the noncompliance rate of concurrent chemoradiation (CCRT), induction of chemotherapy (IC) followed by CCRT and CCRT followed by adjuvant chemotherapy (AC) was 7.02%, 39.76% and 50.00%, respectively. Univariate analysis indicated that NCCN noncompliance regarding the treatment for stage II-IVB NPC did not exhibit a significant correlation with the parameters age, gender, insurance status, education profile, first clinic department, careers, comorbidities and overall clinical stage, but it indicated a significant association with the therapeutic schedule (P<0.05). The multivariate analysis indicated that the NCCN noncompliance regarding the treatment for stage II­IVB NPC exhibited a statistically significant difference between CCRT and CCRT followed by AC (OR=0.10, 95% CI 0.04-0.27, P<0.05 ), although the difference noted between CCRT and IC followed by CCRT was not significantly different (OR=1.71, 95% CI 0.50-5.87,P=0.40). Conclusions: The use of specific therapeutic schedules may affect the noncompliance with NCCN guidelines regarding the treatment for stage II­IVB NPC in southern China, notably with regard to the treatment schedule of CCRT followed by AC.

JNK inhibitor SP600125 enhances TGF-ß-induced apoptosis of RBE human cholangiocarcinoma cells in a Smad-dependent manner.

Transforming growth factor ß (TGF-ß) signaling is pivotal for the progression of specific types of tumors at certain stages. However, the mechanism by which TGF-ß is regulated by other factors remains unclear. In this study, the involvement of SP600125, an inhibitor of c-Jun N-terminal kinase (JNK), in TGF-ß-induced apoptosis of the RBE human cholangiocarcinoma cell line was investigated. Exogenous TGF-ß1 activated Smad and non­Smad signaling pathways, including the JNK pathway in RBE cells, and induced apoptosis, which was inhibited by knockdown of Smad4 expression. SP600125 increased the TGF-ß1­induced phosphorylation of Smad2 and Smad3, which enhanced the TGF-ß1­induced transcriptional response and apoptosis in RBE cells. The effect of SP600125 on the transcriptional response and apoptosis was reduced by knockdown of Smad4 expression. In addition, TGF-ß1­induced apoptosis was abrogated using the pan-caspase inhibitor Z­VAD-fmk. SP600125 promoted the TGF-ß1­induced caspase cleavage, while knockdown of Smad4 expression counteracted this effect. These results indicate that SP600125 enhances TGF-ß-induced apoptosis of RBE cells through a Smad­dependent pathway that involves Smad­dependent caspase activation. SP600125 is hypothesized to be an ideal therapeutic candidate for treating human cholangiocarcinoma.

CD4-CD8-T cells contribute to the persistence of viral hepatitis by striking a delicate balance in immune modulation.

Viral hepatitis remains the most common cause of liver disease and a major public health problem. Here, we focus on the role of CD4 CD8 double negative T (DN T) cells involved in the mechanisms of viral persistence in hepatitis. C3H/HeJ mice infected with murine hepatitis virus strain 3 (MHV-3) were used to display chronic viral hepatitis. DN T cells dramatically increased in MHV-3 infected mice. Adoptive transfer of DN T cells from MHV-3 infected mice led to a significant increase in mice survival. The DN T cells with production of IFN-γ and IL-2 are able to kill virus-specific CD8(+) T cells via the Fas/FasL dependent pathway. The delicate balance of multiple effects of DN T cells may lead to viral persistence in MHV-3 induced hepatitis. In short, our study identified DN T cells contributing to viral persistence in MHV-3 induced hepatitis in C3H/HeJ mice, which provides a rationale for modulating DN T cells for the management of viral hepatitis.

Contribution of IL-17 to mouse hepatitis virus strain 3-induced acute liver failure.

Recently, the Th17 cells and IL-17 have been shown to play a critical role in the immune-mediated liver injury in hepatitis B, while their functions in acute liver failure have not been well elucidated yet. In this study, we primarily investigated the role of IL-17 in the development of mouse hepatitis virus strain 3 (MHV-3)-induced acute liver failure. IL-17 mRNA levels in liver tissue were quantified by using quantitative real-time polymerase chain reaction, and cytokine IL-17 levels in liver tissue and serum were determined by using ELISA in MHV-3-induced murine fulminant hepatitis model. The IL-17 expression levels on CD4(+)T and CD8(+)T cells were determined by using flow cytometry. The correlation between IL-17 level and liver injury was studied. Th17 associated cytokines were also investigated by intracellular staining. Our results showed that the IL-17 expression was significantly elevated in the liver and serum of BALB/cJ mice infected with MHV-3. Moreover, a time course study showed that the percentage of both IL-17-producing CD4(+)T cells and IL-17-producing CD8(+)T cells was increased remarkably in the liver starting from 48 h and peaked at 72 h post-infection. There was a close correlation between hepatic or serum IL-17 concentration and the severity of liver injury defined by ALT level, respectively. Th17 associated cytokines, IL-6, IL-21 and IL-22, were also increased significantly at 72 h post-infection. It was concluded that IL-17 may contribute to the pathogenesis of MHV-3-induced acute liver failure.

Liver TCRγδ(+) CD3(+) CD4(-) CD8(-) T cells contribute to murine hepatitis virus strain 3-induced hepatic injury through a TNF-α-dependent pathway.

The mechanisms of each subset of immune cells contributing to the pathogenesis of viral hepatitis remain incompletely understood. In this study, we examined the role of liver CD4(-) CD8(-) (double negative, DN) T cells during murine hepatitis virus strain 3 (MHV-3)-induced hepatitis in C3H/HeJ mice. We demonstrate that predominant population of DN T cells in the liver of healthy or MHV-3-infected mice express TCRγδ(+). The proportion of TCRγδ(+) DN T cells in liver CD3(+) T cells was markedly increased after MHV-3 infection. Adoptive transfer of TCRγδ(+) DN T cells led to dramatically decreased survival in MHV-3-infected mice, accompanied by deteriorated histopathology and elevated ALT and AST levels. It was found that these cells were hyperactivated after MHV-3 infection with a production of TNF-α, IFN-γ, IL-2 and IL-17A. Highly activated liver TCRγδ(+) DN T cells were cytotoxic to MHV-3-infected hepatocytes in vitro and this effect did not require cell-cell contact. Moreover, the cytotoxic effect of liver TCRγδ(+) DN T cells against hepatocytes involves TNF-α pathway, but not IL-17A or IFN-γ. These results indicate that liver TCRγδ(+) DN T cells play a critical role in the liver injury in MHV-3-induced hepatitis, via a TNF-α dependent pathway.