Proteasomal inhibitors induce myeloma cell pyroptosis via the BAX/GSDME pathway.

Proteasomes are overexpressed in multiple myeloma (MM) and proteasomal inhibitors (PIs) have been widely used for the treatment of MM. PIs are reported to induce MM cell apoptosis but impair necroptosis. In the present study, we found that PIs MG132 and bortezomib induce MM cell pyroptosis, a novel type of cell death, in a GSDME-dependent manner. Lack of GSDME totally blocks PI-induced pyroptosis. Interestingly, we found that Caspase-3/6/7/9 are all involved in pyroptosis triggered by PIs because the specific inhibitor of each caspase ablates GSDME activation. PIs markedly reduce mitochondrial membrane potential. Moreover, PIs disrupt the interaction of Bcl-2 and BAX, induce cytochrome c release from mitochondria to cytosol and activate GSDME. Furthermore, we found that overexpression of an N-terminal portion of GSDME suffices to release cytochrome c from mitochondria and to activate Caspase-3/9, suggesting N-GSDME might penetrate the mitochondrial membrane. Consistent with Bcl-2 inhibition, BAX can induce MM cell pyroptosis in a GSDME-dependent manner. In accordance with these findings, inhibition of Bcl-2 synergizes with PIs to induce MM cell pyroptosis. Therefore, the present study indicates that PIs trigger MM cell pyroptosis via the mitochondrial BAX/GSDME pathway and provides a rationale for combined treatment of MM with Bcl-2 and proteasome inhibitors to increase therapeutic efficiency via induction of pyroptosis.

Risk-adapted stereotactic body radiotherapy for patients with cervical spinal metastases.

Owing to the complex anatomical structure and biomechanics, the current standard palliative treatments for cervical spinal metastases are associated with a high risk of recurrence and complications. Stereotactic body radiotherapy (SBRT) can provide radical dose to tumors while protecting normal organs to the maximum extent. However, the efficacy and safety of SBRT for cervical spinal metastases is not well characterized. Data from 71 patients with cervical spine metastases who were treated with SBRT using CyberKnife between 2006 and 2021 were obtained from our prospectively maintained database. Primary endpoint was pain response at 12 weeks following SBRT completion; secondary endpoints included local control (LC), overall survival (OS), and adverse events. Standard-risk patients were planned to receive 30 Gy (range 21-36) with median fractions of 3 (range 1-3) and high-risk patients 35 Gy (range 24-50) with median fractions of 5 (range 4-5) according to the spinal cord and esophagus dose constraints. The median follow-up time was 17.07 months (range 3.1-118.9). After 12 weeks of SBRT completion, 54 (98.2%) of 55 patients with baseline pain achieved pain response and 46 (83.6%) achieved complete pain response. LC rates were 93.1% and 90% at 1 year and 2 year, respectively. The 1-year and 2-year OS rates were 66.2% and 37.4%, respectively. Eight patients experienced grades 1-4 adverse events (six vertebral compression fracture [VCF], five of them had VCF before SBRT; and two hemiparesis). No grade 5 adverse events were observed. Therefore, risk-adapted SBRT for cervical spine metastases achieved high pain control and LC rates with acceptable adverse events.

Early postoperative radiotherapy is associated with improved outcomes over late postoperative radiotherapy in the management of completely resected (R0) Stage IIIA-N2 non-small cell lung cancer.

AIMS: The aim of this study was to evaluate the ideal timing of PORT in the management of completely resected (R0) Stage IIIA-N2 NSCLC. PATIENTS AND METHODS: Between January 2008 and December 2015, patients with known histologies of pathologic Stage IIIA-N2 NSCLC who underwent R0 resection and received PORT concurrent with or prior to two sequential cycles of chemotherapy ("early PORT") or with PORT administered after two cycles of chemotherapy ("late PORT") at multiple hospitals. The primary endpoint was OS; secondary end points included pattern of the first failure, LRRFS, and DMFS. Kaplan-Meier OS, LRRFS, and DMFS curves were compared with the log-rank test. Cox regression analysis was used to determine prognosticators for OS, LRRFS, and DMFS. RESULTS: Of 112 included patients, 41 (36.6%) and 71 (63.4%) patients received early PORT and late PORT, respectively. The median OS, LRRFS, and DMFS were longer for those who received early PORT than for those who received late PORT at the median follow-up of 29.6 months (all p < 0.05). Uni- and multi-variate analyses showed that number of POCT cycles and the combination schedule of PORT and POCT were independent prognostic factors for OS, LRRFS, and DMFS. CONCLUSIONS: Early PORT is associated with improved outcomes in pathologic Stage IIIA-N2 R0 NSCLC patients.

Necroptosis as a potential therapeutic target in multiple organ dysfunction syndrome.

PURPOSE: To investigate how necroptosisis, i.e. programmed necrosis, is involved in MODS, and to examine whether Nec-1, a specific necroptosis inhibitor, ameliorates multiorgan injury in MODS. EXPERIMENTAL DESIGN: A model of MODS was established in six-week old SD rats using fracture trauma followed by hemorrhage. Control animals received sham surgery. Cell death form and necrosome formation were measured by fluorescence-activated cell sorting and western blotting. MODS rats were randomly assigned to receive Nec-1 or saline with pretreatment and once daily. The first end-point was 72 hours survival. Organ injury and dysfunction, inflammatory cytokine levels, and necroptotic execution protein expression were also recorded. RESULTS: Organ injury and dysfunction were significantly more severe in the MODS group than the sham group (all p<0.01). Furthermore, MODS-induced liver, lung and kidney tissue injury was characterized by necroptosis rather than apoptosis, and accompanied by necrosome formation. Compared to MODS group, Nec-1 administration significantly improved 72 hours survival (p<0.01). Nec-1 administration significantly reduced necroptosis-induced liver, lung and kidney injury and dysfunction, inhibited inflammatory cytokines production, inhibited release of necroptotic execution proteins such as high-mobility group box 1 and mixed-lineage kinase domain-like protein pseudokinase in MODS rats (all p<0.01). CONCLUSIONS: These results suggest that necroptosis is involved the pathology of MODS. Further, a necroptotic inhibitor Nec-1 may be considered as an adjunct treatment for MODS.

Necroptosis in tumorigenesis, activation of anti-tumor immunity, and cancer therapy.

While the mechanisms underlying apoptosis and autophagy have been well characterized over recent decades, another regulated cell death event, necroptosis, remains poorly understood. Elucidating the signaling networks involved in the regulation of necroptosis may allow this form of regulated cell death to be exploited for diagnosis and treatment of cancer, and will contribute to the understanding of the complex tumor microenvironment. In this review, we have summarized the mechanisms and regulation of necroptosis, the converging and diverging features of necroptosis in tumorigenesis, activation of anti-tumor immunity, and cancer therapy, as well as attempts to exploit this newly gained knowledge to provide therapeutics for cancer.

Interleukin-18 and miR-130a in severe sepsis patients with thrombocytopenia.

BACKGROUND: Thrombocytopenia is one of the most common laboratory abnormalities encountered in patients with severe sepsis. It has been reported that thrombocytopenia is linked to mortality in patients with severe sepsis. However, the mechanism of thrombocytopenia in sepsis is unknown. We hypothesized that inflammatory cytokines and microRNAs (miRNAs) are not only involved in the pathogenesis of sepsis, but also are correlated with thrombocytopenia. PATIENTS AND METHODS: Eligible patients with severe sepsis were prospectively recruited and treated at our hospital between June 2012 and May 2014. The miRNA and protein expression of interleukin (IL)-18 and IL-27 were detected by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The expression of miR-130a and miR-150 was detected by TaqMan real-time polymerase chain reaction. RESULTS: Sixty eligible patients were divided into two groups: 28 severe sepsis patients with thrombocytopenia and 32 severe sepsis patients without thrombocytopenia. The results demonstrated that the miRNA expression and plasma concentration of IL-18 in severe sepsis patients with thrombocytopenia were higher than those in severe sepsis patients without thrombocytopenia (P=0.015 and P=0.034, respectively), and miR-130a expression was significantly lower in severe sepsis patients with thrombocytopenia (P<0.003). CONCLUSION: Our data demonstrate that severe sepsis patients with thrombocytopenia have increased plasma and miRNA expression levels of IL-18 and decreased expression of miR-130a, suggesting that IL-18 and miR-130a might be involved in the pathophysiological process of severe sepsis with thrombocytopenia.

Mesenchymal stem cells generate pericytes to promote tumor recurrence via vasculogenesis after stereotactic body radiation therapy.

BACKGROUND: Stereotactic body radiation therapy (SBRT) is postulated to enhance the recruitment of mesenchymal stem cells (MSCs) into the tumor microenvironment, which promote tumor recurrence. The aim of this study is to determine the molecular mechanisms behind SBRT stimulating MSC migration and differentiation. METHODS: In vitro, mediated factors and migrated MSCs (post-SBRT) were generated. In vivo, bone-marrow derived MSCs were identified and harvested from green fluorescent protein (GFP)-expressing transgenic male mice and transplanted into sub-lethally irradiated recipient female mice to establish a model of bone marrow transplantation. Lewis lung carcinoma and malignant melanoma-bearing recipient mice were treated with SBRT, 14 Gy/1 fraction. The migration and differentiation potential of MSCs were characterized. RESULTS: SBRT increased the release of stromal cell derived factor-1α (SDF-1α) and platelet-derived growth factor-B (PDGF-B) by tumor cells; these ligands bound to chemokine (C-X-C motif) receptor 4 (CXCR4) and platelet-derived growth factor receptor-ß (PDGFR-ß), respectively, on circulating bone marrow-derived MSCs, resulting in engraftment of the MSCs into the tumor parenchyma. The newly-homed MSCs differentiated into pericytes, which induced the tumor vasculogenesis, and promoted tumor regrowth. Targeted therapies, AMD3100 and imatinib abrogated MSC homing, vasculogenesis, and tumor regrowth. CONCLUSION: Bone-marrow derived MSCs migrate to the tumor parenchyma and differentiate into pericytes, inducing tumor vasculogenesis after SBRT, and promoting tumor recurrence. MSC migration and maturation may be abrogated with AMD3100 and imatinib. This novel treatment strategy warrants clinical investigation.

Correlation between the suppressor of cytokine signaling-1 and 3 and hepatitis B virus: possible roles in the resistance to interferon treatment.

BACKGROUND: The suppressor of cytokine signaling family (SOCS) is an important negative regulator in the JAK-STAT signaling pathway. This study was designed to explore the correlation between SOCS-1, 2 and 3, Hepatitis B Virus (HBV) and interferon (IFN), and the relationship between SOCS and IFN therapeutic efficacy. METHODS: Four types of mouse models were established. Mice were administered with HBV replicative plasmid pHBV4.1 and IFN inducer Poly IC (Group A), pHBV4.1 (Group B), Poly IC (Group C) and saline (Group D), respectively. Liver tissues were harvested from the mice and SOCS expression was determined. Meanwhile, patients with chronic hepatitis B (CHB) were treated with pegylated interferon α-2b for 24-48 weeks. Liver biopsy was collected and the baseline SOCS expression was determined. Serum assay was performed for efficacy evaluation and correlation analysis. RESULTS: In animal studies, the expression level of SOCS-1 and 3 was found in the descending order of B, A, C and D. The difference between Group B and D suggested that HBV could induce SOCS. The difference between Group A and C suggested that HBV could still induce SOCS with up-regulated endogenous IFN. The difference between Group C and D suggested that ploy IC could induce SOCS, while the difference between Group B and A suggested that Poly IC might have a stronger inhibition effect for SOCS. There was no difference in SOCS-2 expression. In clinical studies, eight of twenty-four enrolled patients achieved either complete or partial therapeutic response. The expression of both SOCS-1 and 3 was higher in CHB patients than in normal controls. The baseline HBV-DNA level was positively correlated with SOCS-1 and 3. The age, viral genotype, HBVDNA, SOCS-1 and SOCS-3 were found to be related to IFN efficacy. CONCLUSION: HBV could induce both SOCS-1 and 3 expression regardless of endogenous IFN level. Elevated IFN could directly up-regulate SOCS-1 and 3 expression, but it could also indirectly down-regulate SOCS-1 and 3 expression by inhibiting HBV replication. HBV might play a more important role in the SOCS up-regulation than IFN, a possible reason why patients with high HBV viral load encounter poor efficacy of IFN treatment.

Meta-analysis: traditional Chinese medicine for improving immune response in patients with unresectable hepatocellular carcinoma after transcatheter arterial chemoembolization.

OBJECTIVE: The aim of this study was to evaluate whether Traditional Chinese Medicine (TCM) improves immune response for unresectable hepatocellular carcinoma (UHCC) after transcatheter arterial chemoembolization (TACE) by using meta-analysis of data from the literature involving available randomized controlled trials of TCM in combination with TACE compared with that of TACE alone. METHODS: Literature retrieval was conducted through the Cochrane Library, MEDLINE, CENTRAL, Embase, CBMdisc, and CNKI, without language limitations. RESULTS: Based on our search criteria, we found 12 trials involving 1,008 patients. Our results showed that the differences of pooled weighted mean difference before and after treatment and 95% confidence intervals (CIs) were 13.63 (8.96-18.69; P = .0001) for the proportion of CD3(+) T cells, 10.56 (6.91-14.21; P = .0001) for the proportion of CD4(+) T cells, -3.40 (-6.83 to 0.03; P = .052) for the proportion of CD8(+) T cells, 0.54 (0.42-0.66; P = .0001) for the ratio of CD4(+)/CD8(+), and 12.34 (7.26-17.41; P = .0001) for the proportion of natural killer cells. No serious adverse events were reported. CONCLUSIONS: Traditional Chinese Medicine in combination with TACE improves the immune response of patients with UHCC. However, considering the strength of the evidence, additional randomized controlled trials are needed before TCM plus TACE can be recommended routinely.

Nucleoside analogue can improve the long-term prognosis of patients with hepatitis B virus infection-associated acute on chronic liver failure.

BACKGROUND: The prognosis of patients with hepatitis B virus (HBV)-associated acute on chronic liver failure (ACLF) is extremely poor. AIM: This study was designed to evaluate the efficacy and safety of nucleoside analogue treatment of patients with HBV-associated ACLF. METHODS: We used a retrospective review of eligible patients from April 2006 to December 2008. Eligible subjects received 0.5 mg entecavir daily until October 2009 (group A), 100 mg lamivudine daily until October 2009 (group B), or no nucleoside analogue (group C). The primary endpoints were three-month survival and the rate of recurrence of HBV-associated ACLF. The secondary endpoints were HBV DNA levels, liver function, the model of end-stage liver disease (MELD) score, and adverse events. RESULTS: A total of 104 consecutive patients were recruited, and 33, 34, and 37 patients were randomly allocated to groups A, B, and C, respectively. Although no significant difference in three-month survival was observed, levels of HBV DNA and rates of recurrence of HBV-associated ACLF were lower. Liver function and MELD score were not significantly improved despite significantly reduced HBV DNA levels. CONCLUSIONS: These data indicated that nucleoside analogue treatment did not improve the short-term prognosis of patients with HBV-associated ACLF although it was efficacious and safe in the management of HBV DNA levels. Intriguingly and importantly, continuous nucleoside analogue treatment can significantly reduce the rate of recurrence, which might be indicative of the further benefit of long-term survival.

Mixed microencapsulation of rat primary hepatocytes and Sertoli cells improves the metabolic function in a D-galactosamine and lipopolysaccharide-induced rat model of acute liver failure.

Hepatocyte transplantation is an alternative to transplantation of the whole liver. Compared with xenogeneic hepatocytes, primary hepatocytes have some advantages, such as a more powerful function and a smaller frequency of rejection caused by the host. Cell microencapsulation prevents direct access of host cells to the graft but cannot impede transfer of transplant-derived peptides, which can cross the physical barrier. Sertoli cells are central to the immune privilege demonstrated in the testis, and their actions have been utilized to protect cell transplants. Co-microencapsulating Sertoli cells with HepG2 cells has proved to be a valuable strategy in hepatocyte transplantation. Thus mixed microcapsules of primary rat hepatocytes and primary Sertoli cells may improve metabolic function in a d-galactosamine and lipopolysaccharide-induced rat model of acute liver failure.

Transcatheter arterial chemoembolization in combination with radiotherapy for unresectable hepatocellular carcinoma: a systematic review and meta-analysis.

BACKGROUND AND PURPOSE: To evaluate the efficacy and safety of transcatheter arterial chemoembolization (TACE) plus radiotherapy (RT) for unresectable hepatocellular carcinoma (UHCC) using meta-analysis of data from the literature involving available randomized controlled trials of TACE in combination with RT compared with that of TACE alone (Therapy I versus II) in treating UHCC. MATERIAL AND METHODS: We searched the Cochrane Library, MEDLINE, CENTRAL, EMBASE, CBMdisc, and CNKI as well as employing manual searches. Meta-analysis was performed on the results of homogeneous studies. Analyses subdivided by study design were also performed. RESULTS: We found 17 trials involving 1476 patients. 5 of total were Randomized Controlled Trials (RCTs) and 12 were Non-randomized Controlled Clinical Trials (CCTs). In terms of quality, 5 RCTs were graded B, and 12 CCTs were graded C. Our results showed that Therapy I, compared with Therapy II, significantly improved the survival and the tumor response of patients, and was thus more therapeutically beneficial. Serious adverse events were not increased exception for total bilirubin (TB) level. CONCLUSIONS: Therapy I was more therapeutically beneficial. However, considering the strength of the evidence, additional randomized controlled trials are needed before Therapy I can be recommended routinely.

Traditional Chinese medicine plus transcatheter arterial chemoembolization for unresectable hepatocellular carcinoma.

OBJECTIVES: To compare the efficacy and safety of Traditional Chinese Medicine (TCM) plus transcatheter arterial chemoembolization (TACE) with that of TACE alone (therapy I versus therapy II, respectively) in treating unresectable hepatocellular carcinoma (UHCC) through a meta-analysis of all available randomized controlled trials. METHODS: Literature retrieval was conducted using the Cochrane Library, MEDLINE, EMBASE, CBMdisk, and CNKI in any language. Meta-analysis was performed on the results of homogeneous studies. Analyses subdivided by TACE frequency (subgroup A, <3 times; subgroup B, > or =3 times) were also performed, but were not done for both therapy I and therapy II. RESULTS: Based on our search criteria, we found 37 trials involving 2653 patients. Our results showed that therapy I, compared with therapy II, improved patient survival, quality of life, alleviation of symptoms, and tumor response, and was thus more therapeutically beneficial. Further analysis showed that subgroup A proved to be better for patients' survival and alleviation of symptoms, while the two subgroups were similar in improved tumor response. No serious adverse events were reported. CONCLUSIONS: Therapy I benefited patients with UHCC. Subgroup A improved the survival of patients and the amelioration of symptoms more than subgroup B. As in some trials, there were flaws in the methodological quality, and the data therefore have a risk of bias and of being insufficient for determining the effects of therapy I and subgroup A. Hence, further large-scale trials are warranted.

Recombinant human hepatocyte growth factor for liver failure.

The purpose of this paper is to evaluate the efficacy and safety of recombinant human hepatocyte growth factor (rh-HGF) for liver failure (LF) using meta-analysis of data from the literature involving available randomized controlled trials of rh-HGF plus comprehensive therapy (CT) compared with that of CT alone (Therapy I versus II) in treating LF. We searched the Cochrane Library, MEDLINE, EMBASE, CBMdisc, and CNKI as well as employing manual searches. Based on our search criteria, we found 21 trials, involving 5902 patients. Our results showed that Therapy I, compared with therapy II, significantly reduced the overall mortality (RR=0.62; 95% CI, 0.59-0.66; p=0.0001). Compared to two clinical types of LF (acute and acute-on-chronic), therapy I perhaps had significant effect on mortality due to sub-acute LF, RR and 95% CI were 0.76 [0.65, 0.89], 0.66 [0.60, 0.74], and 0.58 [0.53, 0.64], respectively. Additionally, there was a reduction in mortality of patients that had evidence for an early stage of LF compared to the two other clinical stages of LF (Middle and Advanced); RR and 95% CI were 0.34 [0.24, 0.49], 0.49 [0.44, 0.55], and 0.87 [0.82, 0.93], respectively. No serious adverse events were reported. We conclude that Therapy I may reduce mortality in LF, especially in sub-acute LF and the early stage of LF. However, considering the strength of the evidence, additional randomized controlled trials are needed before Therapy I can be recommended routinely.