Muramyl dipeptide causes mitochondrial dysfunction and intestinal inflammatory cytokine responses in rats.

INTRODUCTION: Intestinal flora and the translocation of its products, such as muramyl dipeptide (MDP), are common causes of sepsis. MDP is a common activator of the intracellular pattern recognition receptor NOD2, and MDP translocation can cause inflammatory damage to the small intestine and systemic inflammatory responses in rats. Therefore, this study investigated the effects of MDP on the intestinal mucosa and distant organs during sepsis and the role of the NOD2/AMPK/LC3 pathway in MDP-induced mitochondrial dysfunction in the intestinal epithelium. METHODS: Fifty male Sprague Dawley rats were randomly divided into five treatment groups: lipopolysaccharide (LPS) only, 1.5 and 15 mg/kg MDP + LPS, and 1.5 and 15 mg/kg MDP + short-peptide enteral nutrition (SPEN) + LPS. The total caloric intake was the same per group. The rats were euthanized 24 hours after establishing the model, and peripheral blood and small intestinal mucosal and lung tissues were collected. RESULTS: Compared to the LPS group, both MDP + LPS groups had aggravated inflammatory damage to the intestinal mucosal and lung tissues, increased IL-6 and MDP production, increased NOD2 expression, decreased AMPK and LC3 expression, increased mitochondrial reactive oxygen species production, and decreased mitochondrial membrane potential. Compared to the MDP + LPS groups, the MDP + SPEN+LPS groups had decreased IL-6 and MDP production, increased AMPK and LC3 protein expression, and protected mitochondrial and organ functions. CONCLUSIONS: MDP translocation reduced mitochondrial autophagy by regulating the NOD2/AMPK/LC3 pathway, causing mitochondrial dysfunction. SPEN protected against MDP-induced impairment of intestinal epithelial mitochondrial function during sepsis.

Survival Outcomes of Ewing Sarcoma and Rhabdomyosarcoma by High- versus Low-Volume Cancer Centres in British Columbia, Canada.

Due to the rarity and complexity of treatment for Ewing sarcoma and rhabdomyosarcoma, studies demonstrate improved patient outcomes when managed by a multidisciplinary team at high-volume centres (HVCs). Our study explores the difference in outcomes of Ewing sarcoma and rhabdomyosarcoma patients based on the centre of initial consultation in British Columbia, Canada. This retrospective study assessed adults diagnosed with Ewing sarcoma and rhabdomyosarcoma between 1 January 2000 and 31 December 2020 undergoing curative intent therapy in one of five cancer centres across the province. Seventy-seven patients were included, 46 seen at HVCs and 31 at low-volume centres (LVCs). Patients at HVCs were younger (32.1 vs. 40.8 years, p = 0.020) and more likely to receive curative intent radiation (88% vs. 67%, p = 0.047). The time from diagnosis to first chemotherapy was 24 days shorter at HVCs (26 vs. 50 days, p = 0.120). There was no significant difference in overall survival by treatment centre (HR 0.850, 95% CI 0.448-1.614). Variations in care exist amongst patients treated at HVCs vs. LVCs, which may reflect differences in access to resources, clinical specialists, and varying practice patterns across centres. This study can be used to inform decisions regarding triaging and centralization of Ewing sarcoma and rhabdomyosarcoma patient treatment.

Risk of recurrence and pregnancy outcomes in young women with breast cancer who do and do not undergo fertility preservation.

PURPOSE: To assess the impact of fertility preservation (FP) requiring ovarian stimulation on breast cancer outcomes and pregnancy after breast cancer. METHODS: Women aged ≤ 40 years diagnosed with stage I-III breast cancer between 2007 and 2018 and referred for FP consultation prior to systemic therapy were identified from a British Columbia fertility center database. The primary endpoint was invasive breast cancer-free survival (iBCFS) and secondary endpoints were overall survival (OS) and achievement of pregnancy. Survival and pregnancy endpoints were compared using Cox and logistic regression analyses, respectively, for patients who did and did not undergo FP. RESULTS: The study included 153 patients, with 71 (46%) in the FP group and 82 (54%) in the non-FP group. Patients who underwent FP were more likely to be ECOG 0 (99% vs. 88%, p = 0.011) and receive chemotherapy (93% vs. 67%, p < 0.001), but had similar ER positivity status to non-FP patients (70% vs. 79%, p = 0.21). Over a median follow-up of 4.1 years, there were no differences in iBCFS (HR 1.006, 95% CI 0.416-2.438, p = 0.988) or OS (HR 0.789, 95% CI 0.210-2.956, p = 0.725) between FP and non-FP groups. Patients who underwent FP had higher odds of conceiving at least once (OR 3.024, 95% CI 1.312-6.970, p = 0.008). CONCLUSION: At a median follow-up of 4.1 years, FP did not impact iBCFS or OS, supporting its safety in young women with breast cancer. In addition, patients who underwent FP were more likely to become pregnant after breast cancer, highlighting the value of pre-oncologic treatment FP in survivorship family planning.

Impact of GGCX polymorphisms on warfarin dose requirements in atrial fibrillation patients

Background/aim: Warfarin is a common anticoagulant with large interindividual differences and a narrow therapeutic range. The polymorphisms of gamma-glutamyl carboxylase (GGCX) are important genetic factors for warfarin dose requirements. Materials and methods: Polymerase chain reaction and direct sequencing methods were used to detect the GGCX rs699664 genotype in 215 atrial fibrillation (AF) patients with warfarin administration. The effects on warfarin dose by different genotypes were analyzed. A warfarin dosing algorithm was developed based on age, height, CYP2C9, VKORC1, and GGCX genotype. Results: In 215 AF patients, there were 104 cases of wild-type GG genotype (48.4%), 92 cases of GA genotype (42.8%), and 19 cases of AA genotype (8.8%). Patients with the GGCX rs699664 A allele (GA or AA genotypes) needed higher warfarin doses than those with the GG genotype (P < 0.05). A warfarin dosing algorithm showed that age, height, CYP2C9, VKORC1, and GGCX genotype were the best variables for estimating warfarin dose (R2 = 41.2%). Another independent cohort of 60 AF patients showed a significant linear correlation between predicted warfarin maintenance dose and actual dose (R = 0.660, P < 0.01). Conclusion: AF patients with the GA and AA genotypes in GGCX rs699664 required significantly higher warfarin doses. GGCX rs699664 is a potential predictor for the warfarin dose of AF patients.

Anterior mediastinum invasion by multiple myeloma: A case report.

Multiple myeloma (MM) is a clonal proliferation of malignant plasma cells in the bone marrow (BM) that secretes monoclonal paraproteins in the blood serum and urine. Bone marrow MM cells can invade and damage the functions of other tissues and organs, such as the lungs, spleen, liver, pancreas, kidneys and lymph nodes. However, the invasion of MM cells primarily located in the BM to the anterior mediastinum at the site of the thymus is an extremely rare event. The current study reports the case of a 53-year-old female who presented with MM with involvement of the anterior mediastinum. The diagnosis was based on clinical imaging analyses and the results from BM and laboratory examinations, local biopsy pathology and immunohistochemistry. The patient was administered two courses of chemotherapy (epirubicin, dexamethasone and thalidomide). As a result, the tumor reduced in size, but the laboratory examination indicated no significant change. Next, the patient was switched to one course of PAD chemotherapy (bortezomib, epirubicin and dexamethasone). The original tumor was significantly reduced in size following this chemotherapy, and all the indicators improved. The present study suggests that invasion of the thymus by MM may lead to immune disturbance arising from the abnormal thymus gland. In the clinic, extramedullary plasmacytoma in the thymus should be carefully distinguished from thymoma.

DNA damage-induced NF-κB activation in human glioblastoma cells promotes miR-181b expression and cell proliferation.

BACKGROUND: Glioblastoma (GBM) is the most common and most aggressive form of brain cancer. After surgery, radiotherapy is the mainstay of treatment for GBM patients. Unfortunately, the vast majority of GBM patients fail responding to radiotherapy because GBM cells remain highly resistant to radiation. Radiotherapy-induced DNA damage response may correlate with therapeutic resistance. METHODS: Ionizing radiation (IR) was used to induce DNA damage. Cell proliferation and migration were detected by wound-healing, MTT and apoptosis assays. Dual-luciferase assays and Western blot analysis were performed to evaluate NF-κB activation and validate microRNA targets. Real-time PCR was used to study mRNA and microRNA levels. RESULTS: IR-induced DNA damage activated NF-κB in GBM cells which promoted expression of IL-6, IL-8 and Bcl-xL, thereby contributing to cell survival and invasion. Knockdown SENP2 expression enhanced NF-κB essential modulator (NEMO) SUMOylation and NF-κB activity following IR exposure. miR-181b targets SENP2 and positively regulated NF-κB activity. CONCLUSION: NF-κB activation by DNA damage in GBM cells confers resistance to radiation-induced death.

[Clinical research of safflower injection on hibernating myocardial revascularization].

Coronary artery disease (CAD) is one of the leading causes of death. Safflower attracts great attention owing to its anti-ischemia/reperfusion injury effect. Ninety-three patients with CAD were included and randomized into safflower treatment group, PCI group and control group. Low-dose dobutamine stress echocardiography (DSE) was performed to measure end-systolic volume (ESV), end-diastolic volume (EDV), left ventricular ejection fraction (LVEF) and wall motion score index (WMSI) to determine the recovery of hibernating myocardium and cardiac function in all patients before treatment and after 3-month follow-up. The study was to investigate the effects of safflower on hibernating myocardial revascularization and cardiac function. It was found that LVEF was significantly improved, while the ESV and WMSI were significantly reduced after 2-week treatment in safflower and PCI treatment groups. No significant differences were found between safflower and PCI treatment groups in ESV, EDV, WMSI and LVEF after treatment Safflower injection effectively improved hibernating myocardial function.

ß-Elemene inhibits proliferation through crosstalk between glia maturation factor ß and extracellular signal­regulated kinase 1/2 and impairs drug resistance to temozolomide in glioblastoma cells.

ß-elemene, a plant-derived drug extracted from Curcuma wenyujin, has demonstrated marked antiproliferative effects on glioblastoma, while toxicity remains low. However, the underlying molecular mechanisms of the antitumor activity of ß-elemene remain to be elucidated. Previously, it was identified that the glia maturation factor ß (GMFß)/mitogen-activated protein kinase kinase (MAPK) 3/6/p38 pathway participates in the antiproliferative activity of ß-elemene on glioblastoma. In the present study, in order to illustrate the association of GMFß and the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway, U87 and U251 cells were treated with ß-elemene at various doses and for different durations, and the expression of phosphorylated ERK1/2 (p-ERK1/2), ERK1/2, B-cell lymphoma 2 (Bcl-2), Bcl2-associated X and survivin was examined by western blot analysis. Following treatment with ß-elemene and the ERK1/2 inhibitor PD98059, U87 cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay, and the expression levels of Bcl-2 and survivin were examined by western blot analysis. GMFß was then downregulated by RNA interference in ß-elemene-treated U87 cells, and the effect of this on the expression of ERK1/2 and p-ERK1/2 was determined by western blot analysis. Finally, the chemosensitisation of U87 cells to temozolomide (TMZ) through ß-elemene was examined using the CCK-8 assay. The results demonstrated that ß-elemene inhibited the proliferation of U87 glioblastoma cells through the GMFß­dependent inactivation of the ERK1/2-Bcl-2/survivin pathway. Furthermore, inhibition of ERK1/2 by PD98059 enhanced the antitumor effect of ß-elemene and impaired the expression levels of Bcl-2 and survivin. ß-elemene also increased the sensitivity of U87 glioblastoma cells to the chemotherapeutic TMZ, which was synergistically enhanced by PD98059. In conclusion, these results suggested that GMFß-dependent inactivation of the ERK1/2-Bcl-2/survivin pathway mediated the antiproliferative effect of ß-elemene on glioblastoma. Therefore, ß-elemene is a promising chemosensitizer or adjuvant therapeutic for TMZ against glioblastoma brain tumors.

Microsurgical treatment for parasagittal meningioma in the central gyrus region.

The aim of the present study was to determine the efficacy of microsurgery treatment for parasagittal meningioma in the central gyrus region. A microsurgical technique was used to treat 26 patients with large parasagittal meningioma in the central gyrus region. The Rolandic and draining veins and the peritumoral normal brain tissue were retained, and the associated sagittal sinus was appropriately protected. A Simpson grade I, II or III resection was performed in 8 (30.8%), 12 (46.2%) and 6 (23.1%) patients, respectively, with no post-operative mortalities. Following treatment, 9 patients exhibited hemiparalysis. No tumor recurrence was found in 21 patients during the follow-up examination. The treatment protocol described in the current study included sufficient pre-operative imaging evaluations, a skilled microsurgical technique, improved protection of the Rolandic vein and treatment of the sagittal sinus, and was found to significantly increase the total tumor removal rate and decrease post-operative recurrence.

Β-elemene inhibits Hsp90/Raf-1 molecular complex inducing apoptosis of glioblastoma cells.

ß-Elemene, an active component of herb medicine Curcuma wenyujin, has been shown to antagonize glioblastoma cells by inducing apoptosis. However, how ß-elemene induces apoptosis of these cells remains unclear. In this study, we report that ß-elemene disrupted the formation of the Hsp90/Raf-1 complex, a key step in maintaining the conformation stability of Raf-1, and caused deactivation of Raf-1 and inhibition of the ERK pathway, thereby leading to apoptosis of glioblastoma cells. Specifically, treatment of glioblastoma cell lines with ß-elemene attenuated phosphorylation of multiple members of the kinase families in the Ras/Raf/MEK/ERK cascade, including Raf-1 and ERK as well as downstream signaling targets such as Bcl-2. These results suggest that the Hsp90/Raf-1 complex could be a promising molecular target for new drug development for the treatment of glioblastoma.

Lysosomal membrane permeabilization contributes to elemene emulsion-induced apoptosis in A549 cells.

Elemene is a broad-spectrum antitumor agent. In the present study, lysosomal membrane permeabilization (LMP) was detected after short elemene emulsion--exposure (12 h) that preceded a decrease of the mitochondrial membrane potential and DNA damage (24 h) in A549 cells. At later time points (36 h) elemene emulsion caused the appearance of A549 cells with apoptotic features, including apoptotic morphology, phosphatidylserine exposure, and caspase-3 activation. A significant increase in protein expression for cathepsin D was also observed utilizing Western blot analysis after exposure to elemene emulsion for 12 h. The present study showed that elemene emulsion induced the increased levels of reactive oxygen species (ROS) and depletion of glutathione (GSH) in A549 cells. Cells treated with pepstatin A, an inhibitor for cathepsin D, showed a significant inhibition in DNA damage, mitochondrial membrane permeabilization, caspase-3 activation, and phosphatidylserine exposure. These results demonstrated that apoptosis induced by elemene emulsion in A549 cells is mediated in part through LMP and lysosomal protease cathepsin D.

Anti-tumor effect of beta-elemene in glioblastoma cells depends on p38 MAPK activation.

beta-Elemene, a natural plant drug extracted from Curcuma wenyujin, has been used as an antitumor drug for different tumors, including glioblastoma. However, the mechanism of its anti-tumor effect is largely unknown. Here we report that anti-proliferation of glioblastoma cells induced by beta-elemene was dependent on p38 MAPK activation. Treatment of glioblastoma cell lines with beta-elemene, led to phosphorylation of p38 MAPK, cell-cycle arrest in G0/G1 phase and inhibition of proliferation of these cells. Inhibition of p38 MAPK reversed beta-elemene-mediated anti-proliferation effect. Furthermore, the growth of glioblastoma cell-transplanted tumors in nude mice was inhibited by intraperitoneal injection of beta-elemene. Taken together, our findings indicate that activation of p38 MAPK is critical for the anti-proliferation effect of beta-elemene and that p38 MAPK might be a putative pharmacological target for glioblastoma therapy.

[Effect of p38 MAPK on elemene-induced cell cycle arrest in C6 glioblastoma cells].

OBJECTIVE: To observe the effect of elemene on cell cycle of rat C6 glioblastoma cells. METHODS: Cell cycle analysis and expression of p38 in C6 glioblastoma cells under elemene treatment were measured by flow cytometry and Western blot. Flow cytometry and MTT assay were used to examine cell cycle and cell proliferation while C6 glioblastoma cells were pretreated with p38 inhibitor and DN-p38 plasmids. RESULTS: The fraction of C6 in the G0/G1 phase increased 11%, 6.95%, 19.57% respectively in the presence of 40, 60 and 80 microg/ml elemene. The level of phosphorylated p38 MAPK was greatly increased in a dose and time-dependent manner. Inhibition of p38 MAPK activation with SB203580 and DN-p38 blocked elemene-induced anti-proliferation effect. CONCLUSION: Elemene could induce G0/G1 cell cycle phase arrest of C6 glioblastoma cells through up-regulation of phosphorylated p38.

[Influence of elemene on the expression of Bcl-2 family genes in rat C6 glioma cells].

OBJECTIVE: To observe the effects of elemene on the induction of apoptosis in rat C6 glioma cells and its influence on expression of Bcl-2 family genes. METHODS: Rat C6 glioma cells were cultured. Elemene of the concentrations of 0, 20, 40, 60, and 80 microg/ml were added for 12, 24, 36, 48, and 72 hours respectively. RT-PCR was used to detect the mRNA expression of Bcl-2/Bcl-x/1 genes. Western blotting was used to detect the protein expression of Bcl-2/Bcl-x/1 genes. The apoptosis of the cells was examined by flow cytometry. RESULTS: The cell counts of the 20, 40, 60, and 80 microg/ml elemene groups were 536 +/- 9, 375 +/- 10, 246 +/- 9, and 112 +/- 10/visual field respectively, all significantly lower than that of the 0 microg/ml elemene group (all F = 1292.416, P < 0.05) and the apoptotic rates of the 20, 40, 60, and 80 microg/ml elemene groups were (27 +/- 2)%, (29 +/- 4)%, (32 +/- 3)%, and (35 +/- 5)% respectively with an Ap peak. The protein expression of Bcl-2/Bcl-x/l genes was decreased in the elemene groups dose and time-dependently. The expression of Bax protein was decreased in the elemene groups too, however, not dose and time-dependently. CONCLUSION: Apoptosis caused by elemene may be associated with the down-regulation of Bcl-2/Bcl-x/l genes.