Ferritinophagy induced ferroptosis in the management of cancer.

BACKGROUND: Ferroptosis, a newly form of regulated cell death (RCD), is characterized by iron dyshomeostasis and unrestricted lipid peroxidation. Emerging evidence depicts a pivotal role for ferroptosis in driving some pathological processes, especially in cancer. Triggering ferroptosis can suppress tumor growth and induce an anti-tumor immune response, denoting the therapeutic promises for targeting ferroptosis in the management of cancer. As an autophagic phenomenon, ferritinophagy is critical to induce ferroptosis by degradation of ferritin to release intracellular free iron. Recently, a great deal of effort has gone into designing and developing anti-cancer strategies based on targeting ferritinophagy to induce ferroptosis. CONCLUSION: This review delineates the regulatory mechanism of ferritinophagy firstly and summarizes the role of ferritinophagy-induced ferroptosis in cancer. Moreover, the strategies targeting ferritinophagy to induce ferroptosis are highlighted to unveil the therapeutic value of ferritinophagy as a target to manage cancer. Finally, the future research directions on how to cope with the challenges in developing ferritinophagy promoters into clinical therapeutics are discussed.

Neutrophils as potential therapeutic targets for breast cancer.

Breast cancer (BC) remains the foremost cause of cancer mortality globally, with neutrophils playing a critical role in its pathogenesis. As an essential tumor microenvironment (TME) component, neutrophils are emerging as pivotal factors in BC progression. Growing evidence has proved that neutrophils play a Janus- role in BC by polarizing into the anti-tumor (N1) or pro-tumor (N2) phenotype. Clinical trials are evaluating neutrophil-targeted therapies, including Reparixin (NCT02370238) and Tigatuzumab (NCT01307891); however, their clinical efficacy remains suboptimal. This review summarizes the evidence regarding the close relationship between neutrophils and BC, emphasizing the critical roles of neutrophils in regulating metabolic and immune pathways. Additionally, we summarize the existing therapeutic approaches that target neutrophils, highlighting the challenges, and affirming the rationale for continuing to explore neutrophils as a viable therapeutic target in BC management.

Association of Adipokines with Alzheimer's Disease in a Chinese Cohort.

BACKGROUND: The correlation between plasma adipose factor levels and Alzheimer's patients is not entirely clear. OBJECTIVE: We aimed to investigate associations between AD and plasma levels of three adipokines including plasma adiponectin, leptin, and resistin. METHODS: A single-center, cross-sectional study recruited AD patients (n = 148) and cognitively normal (CN) controls (n = 110). The multivariate logistic regression analysis was applied to determine associations of adiponectin, leptin, and resistin with the presence of AD. The receiver operating characteristic (ROC) analysis was employed to determine the diagnostic power of adiponectin, leptin and resistin for AD. RESULTS: After adjusted for the conventional risk factors, plasma levels of leptin (OR = 0.417, 95% CI: 0.272-0.638, p < 0.0001) and adiponectin (OR = 1.249, 95% CI: 1.151-1.354, p < 0.0001) were associated with the presence of AD. In total participants, the plasma adiponectin level was negatively correlated with MMSE scores (p < 0.0001) and was positively with CDR scores (p < 0.0001) and age (p < 0.0001). The plasma level of leptin was negatively correlated with CDR scores (p < 0.0001) and positively correlated with MMSE scores (p < 0.0001). Both adiponectin (p < 0. 0001) and leptin (p < 0. 0001) featured higher AUC than the random chance. CONCLUSIONS: Plasma adiponectin and leptin were associated with the presence, symptomatic severity, and diagnostic power of AD, suggesting a potential role of adipokines in the pathogenesis of AD.

Multi-component immune knockout: A strategy for studying the effective components of traditional Chinese medicine.

Periploca forrestii Schltr., a traditional Chinese medicine (TCM), is commonly used to treat autoimmune diseases such as rheumatoid arthritis (RA). However, its mechanism, involving a variety of cardiac glycosides, remains largely unknown. The immune knockout strategy can highly selectively deplete target components by immunoaffinity chromatography (IAC). We aimed to identify the common structural features of cardiac glycosides in P. forrestii and design IAC to specifically recognize these features to achieve the multi-component knockout of potential active substances from the extracts of P. forrestii. A content detection experiment confirmed that the content of a compound with periplogenin structure (CPS) in the extract of P. forrestii was reduced by 45% by IAC of periplogenin. The immunosuppressive ability of the extract on H9 human T lymphocytic cells was weakened after CPS knockout from P. forrestii extract. Molecular biology experiments showed that mRNA expression of interferon-γ (IFN-γ), interleukin-2 (IL-2), and interleukin-6 (IL-6) in H9 cells was up-regulated after CPS knockout, while no significant changes in the expression of interleukin-4 (IL-4) were found. CPS knockout from P. forrestii extract did not cause significant changes in the proliferation of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells incubated with this extract. These results indicate that CPS exhibited immunosuppressive effects via inhibiting the T helper 1 (Th1) cell immune response and not via the anti-inflammatory components in P. forrestii. This is the first use of IAC to achieve multi-component knockout in TCM extracts for identifying effective compounds. This method is effective and reliable and warrants further exploration.

Detecting Critical Functional Ingredients Group and Mechanism of Xuebijing Injection in Treating Sepsis.

Sepsis is a systemic inflammatory reaction caused by various infectious or noninfectious factors, which can lead to shock, multiple organ dysfunction syndrome, and death. It is one of the common complications and a main cause of death in critically ill patients. At present, the treatments of sepsis are mainly focused on the controlling of inflammatory response and reduction of various organ function damage, including anti-infection, hormones, mechanical ventilation, nutritional support, and traditional Chinese medicine (TCM). Among them, Xuebijing injection (XBJI) is an important derivative of TCM, which is widely used in clinical research. However, the molecular mechanism of XBJI on sepsis is still not clear. The mechanism of treatment of "bacteria, poison and inflammation" and the effects of multi-ingredient, multi-target, and multi-pathway have still not been clarified. For solving this issue, we designed a new systems pharmacology strategy which combines target genes of XBJI and the pathogenetic genes of sepsis to construct functional response space (FRS). The key response proteins in the FRS were determined by using a novel node importance calculation method and were condensed by a dynamic programming strategy to conduct the critical functional ingredients group (CFIG). The results showed that enriched pathways of key response proteins selected from FRS could cover 95.83% of the enriched pathways of reference targets, which were defined as the intersections of ingredient targets and pathogenetic genes. The targets of the optimized CFIG with 60 ingredients could be enriched into 182 pathways which covered 81.58% of 152 pathways of 1,606 pathogenetic genes. The prediction of CFIG targets showed that the CFIG of XBJI could affect sepsis synergistically through genes such as TAK1, TNF-α, IL-1ß, and MEK1 in the pathways of MAPK, NF-κB, PI3K-AKT, Toll-like receptor, and tumor necrosis factor signaling. Finally, the effects of apigenin, baicalein, and luteolin were evaluated by in vitro experiments and were proved to be effective in reducing the production of intracellular reactive oxygen species in lipopolysaccharide-stimulated RAW264.7 cells, significantly. These results indicate that the novel integrative model can promote reliability and accuracy on depicting the CFIGs in XBJI and figure out a methodological coordinate for simplicity, mechanism analysis, and secondary development of formulas in TCM.

Insulin­receptor substrate 1 protects against injury in endothelial cell models of ox­LDL­induced atherosclerosis by inhibiting ER stress/oxidative stress­mediated apoptosis and activating the Akt/FoxO1 signaling pathway.

Oxidized low­density lipoprotein (ox­LDL)­induced endothelial cell (EC) injury is a risk factor for atherosclerosis. Therefore, the present study aimed to investigate the effects of insulin­receptor substrate 1 (IRS­1) on injury to ox­LDL­exposed ECs. For this purpose, thoracic aorta tissues were isolated from rats and cultured to obtain ECs, which were then identified using immunohistochemical staining. IRS­1 overexpression plasmid (pcDNA3.1­IRS­1) and IRS­1­small interfering RNA were synthesized and transfected into ECs pre­exposed to ox­LDL. MTT and TUNEL assays were performed to evaluate the cell proliferative activity and apoptosis. Intracellular reactive oxygen species (ROS) production was determined by a flow cytometry assay. Reverse transcription­quantitative PCR was conducted to measure the peroxisome proliferator­activated receptor gamma co­activator 1 alpha (Ppargcla), phosphoenolpyruvate carboxykinase 1 (Pck1) and glucose­6­phosphatase catalytic subunit (G6pc) gene transcription levels. Western blot analysis was then performed to determine the IRS­1, forkhead box O1 (FoxO1), phosphorylated (p­)FoxO1, 78­kDa glucose­regulated protein (GRP78), p­eukaryotic translation initiation factor 2A (eIF2α), CHOP, Akt and p­Akt expression levels. Immunofluorescence staining was used to evaluate p­FoxO1 nuclear localization. The results indicated that IRS­1 significantly enhanced the proliferative activity, whereas it inhibited the apoptosis of ECs in a model of ox­LDL­induced atherosclerosis compared with ECs without IRS­1 treatment (P<0.05). IRS­1 significantly decreased the p­FoxO1/FoxO1 ratio compared with ECs without ox­LDL treatment (P<0.05). IRS­1 significantly downregulated the expression of ER stress biomarkers, including GRP78, CHOP and the p­eIF2α/eIF2α ratio in ox­LDL­exposed ECs compared with ECs without ISR­1 treatment (P<0.05). IRS­1 significantly reduced the intracellular ROS levels in the EC models of ox­LDL­induced atherosclerosis compared with ECs without IRS­1 treatment (P<0.05). Moreover, IRS­1 promoted the phosphorylation of Akt in the EC models of ox­LDL­induced atherosclerosis. IRS­1 also significantly suppressed the transcription of atherosclerosis­associated genes in ox­LDL­exposed ECs compared with ECs without IRS­1 treatment (P<0.05). Furthermore, IRS­1 significantly increased the cytoplasmic localization of p­FoxO1 in EC models of ox­LDL­induced atherosclerosis. On the whole, the findings of the present study demonstrate that IRS­1 exerts protective effects in an EC model of ox­LDL­induced atherosclerosis by inhibiting ER stress/oxidative stress­mediated apoptosis and activating the Akt/FoxO1 signaling pathway.

Microwaves as modulators of membrane stability parameters during hepatic cancer.

PURPOSE: The present study explored the potential of microwaves on membrane fluidity changes in diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC), in vivo. METHODS: Rats were segregated into four groups: normal control, DEN-treated, microwave-treated, DEN+microwave-treated. Brush border membranes (BBM) were isolated from the rats and, using the membrane extrinsic fluorophore pyrene, we assessed the viscosities as well as fluidity parameters. RESULTS: DEN treatment resulted in a significant rise in lipid peroxidation (LPO). Reduced glutathione levels (GSH) and the activities of glutathione reductase (GR), glutathione transferase (GST), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were found to be significantly decreased following DEN treatment. On the other hand, microwave treatment in DEN-treated rats resulted in a significant decrease in the levels of lipid peroxidation but caused a significant rise in the levels of GSH as well in the activities of GR, GST, SOD, CAT and GPx. The results further demonstrated a marked decrease in membrane microviscosity following DEN treatment. On the other hand, a significant increase was observed in the excimer/monomer ratio and fluidity parameter of DEN-treated rats when compared to normal control rats. However, the alterations in membrane microviscosity and the fluidity parameters were significantly restored after microwave treatment. CONCLUSION: The study, therefore, concludes that microwave proved quite useful in the modulation of membrane stability parameters following DEN-induced hepatic cancer.

Efficacy of self-made Gengnian decoction on phosphatidylinositol 3-kinases/protein kinase B/mammalian target of rapamycin signaling pathway in perimenopausal rats.

OBJECTIVE: To investigate the efficacy of self-made Gengnian decoction on expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR) in ovarian tissues of perimenopausal rats. They were identified with symptom pattern of kidney-Yang deficiency in terms of Traditional Chinese Medicine. METHODS: Female Sprague-Dawley rats aged 10-12 months were selected. Estrous cycle was observed by vaginal smears of keratinocytes to screen the perimenopausal model rats. The chosen rats were randomly divided into five groups, including perimenopausal model of kidney-Yang deficiency group (24 rats), self-made Gengnian decoction of high-dose group (24 rats), self-made Gengnian decoction of middle-dose group (24 rats), self-made Gengnian decoction of low dose group (24 rats) and tibolone control group (24 rats). In addition, rats aged 4-6 months were selected as young control group. The perimenopausal model rats of kidney-Yang deficiency were prepared by alternative intramuscular injection of hydrocortisone 5 mg·kg－1·d－1 The successfully prepared models in self-made Gengnian decoction of high-dose, middle-dose and low-dose groups and tibolone control group were given self-made Gengnian decoction 26.4, 13.2 and 6.6 mg·kg－1·d－1, and tibolone tablets solvent 0.22 mg·kg－1·d－1, respectively, through intragastric administration. Models group and young control group were given the same dose of normal saline, 1 time a day for 15 consecutive days. 24 h after the last administration, blood and ovarian tissues were collected after anesthesia with 20% ethyl carbamate. The follicles of different levels in ovarian tissue were observed and counted by histopathological hematoxylin-eosin staining. Enzyme linked immunosorbent assay was applied to test insulin-like growth factor-1 (IGF-1) level in the serum of experimental rats. The expression levels of PI3K, phosphorylated-Akt (p-Akt) and phosphorylated-mTOR (p-mTOR) mRNA in ovarian tissue were detected by quantitative real-time polymerase chain reaction. RESULTS: The total follicle counts of perimenopausal model rats with kidney-Yang deficiency were significantly reduced, and the number of follicles (mainly increased in preantral follicles and antral follicles) in perimenopausal model rats with kidney-Yang deficiency was significantly increased after intervention of high and middle doses of Gengnian decoction and tibolone (P < 0.05). Compared with normal rats in young control group, the levels of IGF-1 in serum of perimenopausal rats with kidney-Yang deficiency were significantly decreased (P < 0.01), and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated. The relative expression levels of PI3K, p-Akt, p-mTOR mRNA in ovarian tissues of perimenopausal rats with kidney-Yang deficiency were significantly lower than those of young rats (P < 0.01), and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated (P < 0.05). CONCLUSION: Self-made Gengnian decoction can increase the levels of IGF-1, PI3K, Akt and mTOR mRNA expression in serum.

Relationship between the Methylation of Folic Acid Metabolism-Related Genes and the Incidence and Prognosis of Esophageal Cancer among Ethnic Kazakhs.

Objective: To investigate the relationship between the hypermethylation of folic acid metabolism-related genes and the incidence and prognosis of esophageal cancer among ethnic Kazakhs in Xinjiang (China). Methods: According to the standard of esophageal cancer diagnosis, exclusion and epidemiological investigation of the experimental and control groups. Ion capture immunoassays were used to measure serum folic acid levels, while methylation-specific polymerase chain reaction was used to detect gene promoter methylation levels. Log-rank tests and Cox regression models were used to identify prognostic factors in the patient population. Results: Serum folic acid levels in the experimental (cancer) group were significantly lower than in the control (non-cancer) group (Z = -9.13, P < 0.001). Furthermore, the methylation rates of MTHFR, CBS, MGMT, P16, FHIT, and RASSF1A in the experimental group were significantly higher than in the control group. Multivariate analysis identified depth of tumor invasion, regional lymph node metastasis, tumor-node-metastasis stage, and CBS and RASSF1A gene methylation status as independent prognostic factors; female gender and high serum folic acid levels were favorable prognostic factors. Conclusions: Low serum folic acid level is a risk factor for esophageal cancer among ethic Kazakhs. Moreover, methylation of MTHFR, CBS, MGMT, P16, FHIT, and RASSF1A is closely related to esophageal cancer tumorigenesis.

NcRNAs and Cholangiocarcinoma.

Cholangiocarcinoma (CCA) is the most common primary biliary malignancy with poor prognosis. Less understanding of its etiology and pathogenesis makes the diagnosis and therapy difficult. Recently, accumulating evidences have demonstrated that deregulated expression of non-coding RNAs (ncRNAs) is closely associated with the etiopathogenesis of CCA. NcRNAs which lack open reading frame are a heterogeneous class of transcribed RNA molecules, including microRNAs, long non-coding RNAs and circular RNAs. Several studies have shown ncRNAs dysregulation is a common central event occurring in CCA and has the potential of being therapy targets. Moreover, ncRNAs can be easily detected in cancer tissues and biofluids, representing valuable tools for diagnosis. In this review, we illustrate the role of ncRNA in the CCA and discuss their potential clinical value.

[Advances in macrophage function and its anti-inflammatory and proresolving activity and role in periodontitis development].

Macrophage plays an important role in human innate immune system. It has powerful functions, such as recognition, phagocytosis, and bacteria and foreign body removal. Periodontitis, which is a chronic infectious disease characterized by gum inflammation and bone loss, is a major cause of tooth loss in adults. Several studies demonstrated that periodontal tissue destruction is caused by the host immune response defending against infections. As an important part of host immune response, macrophage is also involved in periodontitis pathogenesis. Recently, anti-inflammatory and proresolving activities of macrophage was discovered. Thus, the complex function of macrophage in the occurrence, development, and resolution of inflammation and its potential role in periodontitis were reviewed.

Origin and Evolution of Allopolyploid Wheatgrass Elymus fibrosus (Schrenk) Tzvelev (Poaceae: Triticeae) Reveals the Effect of Its Origination on Genetic Diversity.

Origin and evolution of tetraploid Elymus fibrosus (Schrenk) Tzvelev were characterized using low-copy nuclear gene Rpb2 (the second largest subunit of RNA polymerase II), and chloroplast region trnL-trnF (spacer between the tRNA Leu (UAA) gene and the tRNA-Phe (GAA) gene). Ten accessions of E. fibrosus along with 19 Elymus species with StH genomic constitution and diploid species in the tribe Triticeae were analyzed. Chloroplast trnL-trnF sequence data suggested that Pseudoroegneria (St genome) was the maternal donor of E. fibrosus. Rpb2 data confirmed the presence of StH genomes in E. fibrosus, and suggested that St and H genomes in E. fibrosus each is more likely originated from single gene pool. Single origin of E. fibrosus might be one of the reasons causing genetic diversity in E. fibrosus lower than those in E. caninus and E. trachycaulus, which have similar ecological preferences and breeding systems with E. fibrosus, and each was originated from multiple sources. Convergent evolution of St and H copy Rpb2 sequences in some accessions of E. fibrosus might have occurred during the evolutionary history of this allotetraploid.

Echocardiographic Characterization of a Murine Model of Hypertrophic Obstructive Cardiomyopathy Induced by Cardiac-specific Overexpression of Epidermal Growth Factor Receptor 2.

Although rare, hypertrophic cardiomyopathy (HCM) with midventricular obstruction is often associated with severe symptoms and complications. None of the existing HCM animal models display this particular phenotype. Our group developed a mouse line that overexpresses the ErbB2 receptor (ErbB2(tg)) in cardiomyocytes; we previously showed that the ErbB2 receptor induces cardiomyocyte hypertrophy, myocyte disarray, and fibrosis compatible with HCM. In the current study, we sought to further echocardiographically characterize the ErbB2(tg) mouse line as a model of HCM. Compared with their wild-type littermates, ErbB2(tg) mice show increased left ventricular (LV) mass, concentric LV hypertrophy, and papillary muscle hypertrophy. This hypertrophy was accompanied by diastolic dysfunction, expressed as reduced E:A ratio, prolonged deceleration time, and elevated E:e' ratio. In addition, ErbB2(tg) mice consistently showed midcavity obstruction with elevated LV gradients, and the flow profile revealed a prolonged pressure increase and a delayed peak, indicating dynamic obstruction. The ejection fraction was increased in ErbB2(tg) mice, due to reduced end-diastolic and end-systolic LV volumes. Furthermore, systolic radial strain and systolic radial strain rate but not systolic circumferential strain and longitudinal strain were decreased in ErbB2(tg) compared with wild-type mice. In conclusion, the phenotype of the ErbB2(tg) mouse model is consistent with midventricular HCM in many important aspects, including massive LV hypertrophy, diastolic dysfunction, and midcavity obstruction. This pattern is unique for a small animal model, suggesting that ErbB2(tg) mice may be well suited for research into the hemodynamics and treatment of this rare form of HCM.

Neuroendocrine carcinoma of the stomach: clinical features and CT findings.

PURPOSE: The purpose of the study is to investigate the computed tomographic characteristic and clinical findings of gastric neuroendocrine carcinoma (G-NEC) to increase awareness of this disease. METHODS: Twenty-two patients with a diagnosis of G-NEC were identified through the PACS of our hospital from August 2010 to November 2014. The clinical data, computed tomography (CT) features, and pathology records were analyzed. RESULTS: Among the 22 patients, 21 were male (95.45%), and 1 was female (4.55%). The mean age was 63.5 years old. Positive rates of neuroendocrine markers were 77.28% for chromogranin A staining, 86.36% for synaptophysin staining. All cases were single lesions including 16 (72.73%) in the gastric fundus, 3 (13.64%) in the gastric body and 1 (4.55%) in the gastric angle. Additionally 2 (9.09%) were found in the gastric antrum. Gastric wall was local thickening in 15 cases, and mass formation in 7 cases, with the stenosis and deformation of the adjacent gastric cavity. The long-axis diameter of the lesions ranged from 1.2 to7.4 cm (mean diameter, 2.47 cm), and the long-axis diameter was <2 cm in 12 case, 2-7.4 cm in 10 cases. The radiodensity values of the lesions were homogeneous density in 15 cases ranging from 22 to 47 HU (mean 34 HU). An ulcer with an irregular base and slightly raised borders located in the stomach was seen in 19 cases. The CT images showed homogeneous enhancement in 15 cases and heterogeneous enhancement in 7 cases. Obvious enhancement was seen in two cases, moderate enhancement was seen in sixteen cases, and mildly enhancement was seen in four cases. The peak value occurred in the arterial phase in 5 cases and the peak value was seen in 17 cases in the portal phase. Eleven lesions invaded the gastric serosa, and lymphatic metastasis was observed in 21 cases, 8 of which were combined with liver metastasis. CT images revealed 2 cases of the liver metastasis had obvious enhancement. CONCLUSION: The CT features regarding location, incidence rates of ulcer and enhancement pattern described in our findings are common in all malignant gastric tumors. Therefore, the diagnosis of G-NEC must be confirmed with pathological test.

The association between infectious burden and Parkinson's disease: A case-control study.

INTRODUCTION: The etiology of Parkinson's disease (PD) remains unclear. The aim of this study was to examine the association between common pathogenic infections and PD. METHODS: Antibody titers to common infectious pathogens including cytomegalovirus (CMV), Epstein Barr virus (EBV)，herpes simplex virus type-1 (HSV-1), Borrelia burgdorferi (B. burgdorferi), Chlamydophila pneumoniae (C. pneumoniae) and Helicobacter pylori (H. pylori) were measured by ELISA in serum of 131 PD patients and 141 normal controls. Infectious burden (IB) was defined as a composite serologic measure of exposure to these common pathogens. RESULTS: Seropositivities toward zero-two, three-four and five-six of these pathogens were found in 11%, 74% and 15% of normal controls while in 4%, 61% and 35% of PD patients, respectively. IB, bacterial burden and viral burden were independently associated with PD. Schwab and England (S&E) scores were negatively correlated with IB in patients with PD. Serum α-synuclein protein levels and inflammatory cytokines (interleukin-1ß and interleukin-6) in individuals with higher IB were also significantly higher. CONCLUSIONS: IB consisting of CMV, EBV, HSV-1, B. burgdorferi, C. pneumoniae and H. pylori is associated with PD. This study supports the role of infection in the etiology of PD.

A novel bispidinone analog induces S­phase cell cycle arrest and apoptosis in HeLa human cervical carcinoma cells.

2,4,6,8-(3)-Tetranitrophenyl-3,7-diazabicyclo[3.3.1]nonan-9-one (B16), a bispidinone analog, was synthesized to investigate its effects on cell viability, the cell cycle, and apoptotic pathways in HeLa human cervical cancer cells. B16 decreased the percentage of viable cells in WST-8 assays, and morphological changes associated with apoptotic cell death were observed, including cell shrinkage and disruption. Annexin V-FITC/PI dual staining assays showed that B16 significantly increased the early apoptosis of HeLa cells after 24 h of treatment. Moreover, DNA content analysis and [3H]-thymidine incorporation assays showed that B16 induced S-phase cell cycle arrest and inhibited DNA replication after 24 h of treatment. Following treatment with 25 µM of B16, an increase in reactive oxygen species and a decrease in mitochondrial membrane potential were observed by flow cytometry. In addition, the expression levels of caspase cascade and Bcl-2 family proteins determined by western blotting suggested that the induction of apoptosis by B16 was associated with a caspase- and mitochondrial-dependent pathway in HeLa cells. In conclusion, B16 induced early apoptosis and S-phase cell cycle arrest in HeLa cells via a caspase- and mitochondrial­dependent pathway.

Maotai ameliorates diethylnitrosamine-initiated hepatocellular carcinoma formation in mice.

Consumption of alcohol is closely related to liver disease, such as hepatic fibrosis or even hepatocellular carcinoma (HCC). However, epidemiological and experimental studies indicated that consumption of Maotai, one of the famous liquors in China, exhibits no significant correlation with hepatic fibrosis or cirrhosis as other beverage sources do. This study detected the relationship of Maotai consumption and HCC development in a diethylnitrosamine (DEN)-initiated HCC animal model. DEN was given to mice at a dose of 100 mg/kg, ip, and 50 mg/kg, ip in the following week. Mice were simultaneously given Maotai or an equal amount of ethanol (53%, 5 ml/kg/day, 5 days/week for up to 35 weeks). At 3-week and 35- week of the experiment, serum and livers were collected for biochemical and histopathological examination of liver injury and incidence of HCC. Real-time RT-PCR, immunohistochemistry and Western blotting were used to examine the expression of metallothionein-1/2 (MT-1/2), NF-E2-related factor 2 (Nrf2), glutamate-cysteine ligase catalytic subunit (GCLC) and modified subunit (GCLM). We identified tissue damage and dysfunction of liver in ethanol + DEN-treated mice, whereas the extent of injury was reduced in Maotai+ DEN -treated mice. Significant Glypican-3(GPC3) expression and precancerous injury or HCC were seen in approximately 50% of mice with ethanol+ DEN, but barely be seen in Maotai + DEN-treated mice. A higher expression of MT-1/2, Nrf2 and GCLC could be seen in Maotai + DEN-treated mice. Thus, Maotai liquor ameliorates the formation of DEN-induced HCC in mice, and the protection mechanism is possibly related with the activation of anti-oxidation factors, such as MTs, Nrf2 and GCLC.

Endoscopy-assisted cerebral falx incision via unilateral approach in treatment of dissymmetric bilateral frontal contusion.

BACKGROUND: Dissymmetric bilateral frontal contusion (DBFC) is relatively frequent in the clinic. In this study, we aimed to investigate the development tendency, clinical features, and treatment experience of DBFC and to summarize out experience in treating these patients via minimally invasive means-endoscopy. METHODS: Over the past 3 years, we have treated a total of 31 patients with DBFC using endoscopy-assisted unilateral cerebral falx incision. We used a 30-degree endoscope to observe the involvement of brain contusion and whether the brain contusions have been cleaned thoroughly. Another 30 patients treated by routine bilateral approach within the same period were taken as controls. RESULTS: Seventeen cases (54.8%) in the unilateral-operation group survived and were in good condition, 8 cases (25.8%) had moderate disability, and 4 cases (12.9%) had severe disability; 1 case (3.2%) was in vegetable state, and 1 case (3.2%) died. Compared with the control group, the GOS score was not significantly different in the unilateral-operation group, but the operation time, blood transfusion volume, length of hospital stay, incidence of mental disorder, and incidence of olfactory nerve injury were greatly reduced in the unilateral-operation group. CONCLUSIONS: Endoscopy-assisted unilateral cerebral falx incision can shorten the operation time and reduce surgical trauma and complications when used for treatment of patients with DBFC.

GSK-3ß activation mediates Nogo-66-induced inhibition of neurite outgrowth in N2a cells.

The axons of the adult mammalian brain and spinal cord fail to regenerate after injury, and it has been suggested that Nogo-66 could prevent CNS axon repair. However, the mechanism of Nogo-66 inhibiting neurite outgrowth remains unknown. Our previous results indicated that protein kinase B (PKB) is involved in the inhibition of the neurite outgrowth by Nogo-66. Glycogen synthase kinase-3ß (GSK-3ß) is implicated in many processes in the nervous system, including differentiation, specification, polarity, plasticity and axon growth. In addition, GSK-3ß is one of the most important molecules downstream of PKB. In the present study, we report on the role of GSK-3ß signaling on Nogo-66-treated mouse neuroblastoma N2a cells. Nogo-66 reduced the phosphorylation of GSK-3ß at Ser9 in N2a cells. In contrast, pretreatment with SB216763, a specific inhibitor of GSK-3ß, resulted in an amelioration of neurite outgrowth by Nogo-66, compared with the Nogo-66 alone group (P<0.05). Moreover, we performed RNA interference experiments to knock down GSK-3ß expression levels in N2a cells via transient transfection of shRNA plasmids. The inhibition of neurite outgrowth by Nogo-66 was subdued in shRNA cells, compared to the non-RNAi cells (P<0.05). Taken together, these data suggest that GSK-3ß is involved in the inhibition by Nogo-66 of neurite outgrowth in N2a cells.