[Sanggenon C inhibits glioblastoma cell migration and invasion by promoting ubiquitination of ß-catenin].

Glioblastoma is a malignant and highly invasive tumor, which requires new approaches to search for chemotherapeutic agents. Sanggenon C (SC) mainly exists in the root bark of white mulberry. Although its anti-tumor effects have been reported in some cancers, the mechanism remains unclear. In this study, we used microscopic observation, transwell assay, and immunofluorescence assay to verify the effect of Sanggenon C on the migration and invasion of glioblastoma cells. We then carried out the gene set enrichment analysis (GESA), real-time qPCR assay and ubiquitination assay to delineate the molecule mechanism by which Sanggenon C affects the migration and invasion ability of glioblastoma. With the addition of Sanggenon C, glioblastoma cells were rounded up, with the migration and invasion ability weakened as verified by transwell assay and immunofluorescence assay. The results of GESA suggested that SC might regulate the expression of genes associated with migration and invasion and affect the activity of Wnt/ß-catenin signaling pathway. Western blotting revealed that Sanggenon C promoted the ubiquitination of ß-catenin to reduce the levels of ß-catenin and its downstream proteins. This was further supported by the results of real-time qPCR analysis of target genes of ß-catenin. Taken together, SC inhibits glioblastoma cell migration and invasion by enhancing ß-catenin ubiquitination. Our work suggests a new direction for the treatment of glioblastoma.

Nonstructural maintenance of chromatin condensin I complex subunit G promotes the progression of glioblastoma by facilitating Poly (ADP-ribose) polymerase 1-mediated E2F1 transactivation.

BACKGROUND: Nonstructural maintenance of chromatin condensin I complex subunit G (NCAPG), also known as non-structural maintenance of chromosomes condensin I complex subunit G, is mitosis-related protein that widely existed in eukaryotic cells. Increasing evidence has demonstrated that aberrant NCAPG expression was strongly associated with various tumors. However, little is known about the function and mechanism of NCAPG in glioblastoma (GBM). METHODS: The expression and prognostic value of NCAPG were detected in the clinical databases and tumor samples. The function effects of NCAPG downregulation or overexpression were evaluated in GBM cell proliferation, migration, invasion, and self-renewal in vitro and in tumor growth in vivo. The molecular mechanism of NCAPG was researched. RESULTS: We identified that NCAPG was upregulated in GBM and associated with poor prognosis. Loss of NCAPG suppressed the progression of GBM cells in vitro and prolonged survival in mouse models of GBM in vivo. Mechanistically, we revealed that NCAPG positively regulated E2F transcription factor 1 (E2F1) pathway activity. By directly interacting with Poly (ADP-ribose) polymerase 1, a co-activator of E2F1, and facilitating the PARP1-E2F1 interaction to activate E2F1 target gene expression. Intriguingly, we also discovered that NCAPG functioned as a downstream target of E2F1, which was proved by the ChIP and Dual-Luciferase results. Comprehensive data mining and immunocytochemistry analysis revealed that NCAPG expression was positively associated with the PARP1/E2F1 signaling axis. CONCLUSIONS: Our findings indicate that NCAPG promotes GBM progression by facilitating PARP1-mediated E2F1 transactivation, suggesting that NCAPG is a potential target for anticancer therapy.

Sanggenon C inhibits cell proliferation and induces apoptosis by regulating the MIB1/DAPK1 axis in glioblastoma.

Sanggenon C (SC), a herbal flavonoid extracted from Cortex Mori, has been mentioned to possess more than one treasured organic properties. However, the molecular mechanism of its anti-tumor impact in glioblastoma (GBM) remains unclear. In this study, we reported that SC displayed a GBM-suppressing impact in vitro and in vivo with no apparent organ toxicity. SC dramatically suppressed cell proliferation-induced cell apoptosis in GBM cells. Mechanistically, we unveiled that SC modulated the protein expression of death associated protain kinase 1 (DAPK1) by controlling the ubiquitination and degradation of DAPK1. Quantitative proteomic and Western blot analyses showed that SC improved DAPK1 protein degradation via decreasing the expression of E3 ubiquitin ligase Mindbomb 1 (MIB1). More importantly, the effects of SC on cell proliferation and apoptosis of GBM cells have been in part reversed through DAPK1 downregulation or MIB1 overexpression, respectively. These results indicated that SC might suppress cell proliferation and induce cell apoptosis by decreasing MIB1-mediated DAPK1 degradation. Furthermore, we found that SC acted synergistically with temozolomide (TMZ), an anti-cancer drug used in GBM, resulting in elevated chemotherapeutic sensitivity of GBM to TMZ. Collectively, our data suggest that SC might be a promising anti-cancer agent for GBM therapy.

Integrin ß2 and ß3: Two plasmatocyte markers deepen our understanding of the development of plasmatocytes in the silkworm Bombyx mori.

Insect hemocytes play important biological roles at developmental stages, metamorphosis, and innate immunity. As one of the most abundant cell types, plasmatocytes can participate in various innate immune responses, especially in encapsulation and node formation. Here, 2 molecular markers of plasmatocytes, consisting of integrin ß2 and ß3, were identified and used to understand the development of plasmatocytes. Plasmatocytes are widely distributed in the hematopoietic system, including circulating hemolymph and hematopoietic organs (HPOs). HPOs constantly release plasmatocytes with high proliferative activity in vitro; removal of HPOs leads to a dramatic reduction in the circulating plasmatocytes, and the remaining plasmatocytes gradually lose their ability to proliferate in vivo. Our results demonstrated that the release of plasmatocytes from HPOs is regulated by insulin-mediated signals and their downstream pathways, including PI3K/Akt and MAPK/Erk signals. The insulin/PI3K/Akt signaling pathway can significantly irritate the hematopoiesis, and its inhibitor LY294002 could inhibit the hemocytes discharged from HPOs. While the insulin/MAPK/Erk signaling pathway plays a negative regulatory role, inhibiting its activity with U0126 can markedly promote the discharge of plasmatocytes from HPOs. Our results indicate that the circulating plasmatocytes are mainly generated and discharged by HPOs. This process is co-regulated by the PI3K/Akt and MAPK/Erk signals in an antagonistic manner to adjust the dynamic balance of the hemocytes. These findings can enhance our understanding of insect hematopoiesis.

The active ingredient (DSH-20) of Salvia miltiorrhiza flower reduces oxidative damage and apoptosis in cardiomyocytes by regulating miR-1.

BACKGROUND: DSH-20, the active ingredient of Salvia miltiorrhiza flower extract, is used to treat cardiovascular diseases. However, its mechanism of action remains unclear. Herein, we investigated the intervention of DSH-20 in H2O2-induced oxidative damage and apoptosis in cardiomyocytes. METHODS AND RESULTS: H2O2 was used to induce oxidative damage and apoptosis in H9c2 cardiomyocytes. Based on concentration gradient studies, we found that 62.5 µg/mL DSH-20 significantly reduced reactive oxygen species and lactate dehydrogenase levels and increased superoxide dismutase levels. DSH-20 also alleviated the apoptosis rate, the changes in mRNA of apoptosis-related genes (Bcl-2, BAX, and Caspase-3) and miR-1 expression. Moreover, transfection of miR-1 mimics aggravated oxidative damage and apoptosis, whereas DSH-20 alleviated these effects. CONCLUSIONS: DSH-20 reduced H2O2-induced oxidative damage and apoptosis in H9c2 cardiomyocytes likely by downregulating miR-1 expression.

ZC3H15 promotes gastric cancer progression by targeting the FBXW7/c-Myc pathway.

Zinc finger CCCH-type containing 15 (ZC3H15), a highly conserved eukaryotic protein, which was associated with several cellular processes and was ubiquitously expressed in various human tissues. Recent studies indicated that ZC3H15 was involved in tumorigenesis and may be a potential biomarker in hepatocellular carcinoma (HCC) and acute myeloid leukemia (AML). However, the biological function and molecular mechanism of ZC3H15 in gastric cancer (GC) have not been studied. In this study, we revealed that ZC3H15 was highly expressed in GC and high ZC3H15 expression was closely linked to poor survival of patients with GC. We found that ZC3H15 promoted cell proliferation, migration, and invasion by increasing c-Myc expression. Next, we found that ZC3H15 could modulate c-Myc protein stability by suppressing the transcription of FBXW7, which was mainly responsible for c-Myc degradation. Moreover, silencing of FBXW7 in ZC3H15-knockdown GC cells could partly abrogate the effects induced by ZC3H15 downregulation. Taken together, our data unearth the important roles of ZC3H15 in GC development and suggest that ZC3H15 may be a potential target for the treatment of GC.

ZC3H15 promotes glioblastoma progression through regulating EGFR stability.

Zinc finger CCCH-type containing 15 (ZC3H15), a highly conserved protein involved in several cellular processes, which was responsible for tumorigenesis and may be a promising marker in myeloid leukemia (AML) and hepatocellular carcinoma (HCC). However, little is known about the biological significance and molecular mechanisms of ZC3H15 in GBM. In this study, we revealed that ZC3H15 was overexpressed in GBM and high ZC3H15 expression was associated with poor survival of patients with GBM. We found that ZC3H15 promoted the proliferation, migration, invasion, and tumorigenesis of GBM cells by activating the EGFR signaling pathway. We also revealed that ZC3H15 reduced EGFR ubiquitination, which was responsible for EGFR protein stabilization. In addition, we demonstrated that ZC3H15 inhibited the transcription of CBL, which was an E3 ubiquitin ligase for EGFR proteasomal degradation. And silencing of CBL could partly abrogate the inhibitory effects on cell proliferation, migration, and invasion of GBM cells induced by ZC3H15 knockdown. Thus, our research revealed the important roles of ZC3H15 in GBM development and provided a brand-new insight for improving the treatment of GBMs.

miR-126 regulates angiogenesis in myocardial ischemia by targeting HIF-1α.

Promoting angiogenesis by targeting various angiogenic regulators has emerged as a new treatment strategy for myocardial ischemia (MI). MicroRNA-126 (miR-126) has been identified as the main regulator of compensatory angiogenesis; however, its role in MI is unclear. A rat MI model and an EA. hy926 endothelial cell hypoxia model were constructed and it was found that miR-126 was highly expressed in both models. The knockdown of HIF-1α expression in EA. hy926 cells in turn downregulated VEGF and CD34 expression and consequently inhibited angiogenesis. MiR-126 inhibitor inhibited EA. hy926 cell migration and tube formation as well as downregulated VEGF and CD34 expression, and these were reversed by transfection of miR-126 mimics. Rescue tests using miR-126 and HIF-1α demonstrated that miR-126-mediated regulation of angiogenesis was dependent on HIF-1α. In summary, miR-126 regulates the occurrence and progression of angiogenesis during MI via HIF-1α and may be a potential new therapeutic target.

Molecular characterization of the prostaglandin E receptor subtypes 2a and 4b and their expression patterns during embryogenesis in zebrafish.

The molecular expression profiles of zebrafish ep2a and ep4b have not been defined to date. Phylogenetic trees of EP2a and EP4b in zebrafish and other species revealed that human EP4 and zebrafish EP4b were more closely related than EP2a. Zebrafish EP2a is a 281 amino acid protein which shares high identity with that of human (43%), mouse (44%), rat (43%), dog (44%), cattle (41%), and chicken (41%). Zebrafish EP4b encoded a 497 amino acid precursor with high amino acid identity to that of mammals, including human (57%), mouse (54%), rat (55%), dog (55%), cattle (56%), and chicken (54%). Whole-mount in situ hybridization revealed that ep2a was robustly expressed in the anterior four somites at the 10-somites stages, but was absent in the somites at 19 hpf. It was observed again in the pronephric duct at 24 hpf, in the intermediate cell mass located in the trunk, and in the rostral blood island at 30 hpf. Ep2a was also expressed in the notochord at 48 hpf. During somitogenesis, ep4b was highly expressed in the eyes, somites, and the trunk neural crest. From 30 to 48 hpf, ep4b could be detected in the posterior cardinal vein and the neighboring inner cell mass. From these data we conclude that ep2a and ep4b are conserved in vertebrates and that the presence of ep2a and ep4b transcripts during developmental stages infers their role during early zebrafish larval development. In addition, the variable expression of the two receptor isoforms was strongly suggestive of divergent roles of molecular regulation.

Retrospective Analysis of 28 Cases Confirmed for Primary Angiitis of the Central Nervous System by Biopsy.

OBJECTIVE: The present study aimed to summarize the clinical characteristics, therapeutic effects, and long-term prognosis of cases confirmed with primary angiitis of the central nervous system (PACNS) by biopsy, analyze the risk factors, and provide clinical guidance for the diagnosis and treatment of the disease. METHODS: Retrospective analysis was performed on 28 cases of PACNS confirmed by biopsy, and the age, gender, pathological results, course of the disease, imaging manifestations, treatment, and prognosis of the patients were analyzed and summarized. RESULTS: The cohort (age 16-60 years) comprised of 16 males. The average time from the visit to diagnosis was 6 months. The first symptom was chronic headache in 18 patients. The pathological results were accompanied by demyelination in 10 cases and glial hyperplasia in 6 cases. A total of 27 patients received treatments including glucocorticoid+cyclophosphamide; of these, 3 cases of craniotomy were improved. Among the 28 patients, 15 patients improved after the treatment, 12 patients had no significant improvement, and 1 patient was deceased. Patients with a long course of the disease before diagnosis, a Karnofsky performance status (KPS) score <60 at the time of diagnosis, a behavioral, cognitive abnormality before treatment, and a short-term relapse (0.3-1 month) have a poor outcome. CONCLUSIONS: PACNS patients are prone to misdiagnosis and mistreatment, with unknown etiology and poor prognosis due to delayed treatment. Therefore, early biopsy, pathological diagnosis, and timely treatment with glucocorticoid shock are recommended, and patients with obvious mass effect should be treated by surgical resection.

Neutrophil-Derived MRP14 Supports Plasma Cell Commitment and Protects Myeloma Cells from Apoptosis.

Neutrophils have recently been proposed as cells with high functional plasticity and are involved in the pathogenesis of infections, malignancy, and autoimmune diseases. However, less is known about the role of neutrophil in humoral response. In this study, we examined the importance of neutrophils and the neutrophil-derived DAMP protein, MRP14, in antibody production. Splenic neutrophils and MRP14 that are present in the splenic peri-MZ region have a close contact with MZ B cells and promote their differentiation into plasma cells. Using neutrophil-depleting mice and an MRP14-blocking compound, we showed that the presence of neutrophil and MRP14 is required for class switch, plasma cell maintenance, and antibody production in the spleen. We found that MRP14 could also be produced by neutrophils in the bone marrow and support the maintenance of bone marrow plasma cells. MRP14 binding could enhance the effect of the BAFF signal and protect primary multiple myeloma cells from doxorubicin-induced apoptosis. Our data demonstrate the effects of neutrophils on neighboring B cells and plasma cells, which provides new insights into the connection between neutrophil and humoral responses.

Drug distribution and clinical safety in treating cystic craniopharyngiomas using intracavitary radiotherapy with phosphorus-32 colloid.

The present study evaluated drug distribution and clinical safety in treating patients with cystic craniopharyngioma (CP) with intracavitary radiotherapy using phosphorus-32 (32P) colloid. In total, 40 patients who were recently diagnosed with primary or recurrent cystic CP were enrolled into the study. Patients underwent stereotactic intracavitary therapy and were administered 32P colloid and iopamidol-300 (1:1 dilution). Head computed tomography (CT) scans were performed 2 h after surgery in order to assess drug distribution and leakage. Results obtained from the ophthalmic examination (visual acuity, visual field and fundus), enhanced head magnetic resonance imaging and/or CT scans, blood analysis, coagulation tests, electrolyte tests, pituitary hormone level analysis, and hepatic and renal function tests were compared between the 0.5, 1, 1.5 and 2 mCi groups. The 32P colloid per minute radioactive count was quantitatively measured in urine and blood samples using a CAPRAC well-type NaI γ counter at 1, 3 and 7 days post-surgery. In total, 6, 2 and 1 case(s) from the 2, 1.5 and 1 mCi groups, respectively, demonstrated heterogeneous drug distribution and intracavitary cerebrospinal fluid leakage. Furthermore, out of 24 patients, no significant differences were identified in blood analysis, blood biochemical measurements and pituitary hormone levels prior to and 7 days after surgery. Blood 32P deposition returned to normal levels within 3 days after surgery, whereas urine deposition returned to normal within 7 days after surgery. Methods utilized in the present study were advantageous in terms of convenience, speed and low cost, therefore, these techniques are suitable for continuous monitoring of patient 32P colloid deposition.

Application research on three-dimensional ultrasonic skeletal imaging mode in detecting fetal upper jaw bone.

OBJECTIVE: To detect three-dimensional (3D) ultrasound appearance of fetal normal and abnormal supermaxilla bone's anatomy using skeletal rendering mode, and to compare the success rate of 3D images in different gestational age groups. METHODS: Using three-dimensional ultrasound skeletal rendering mode of voluson 730 and voluson E8 ultrasound systems, the fetal supermaxilla bones were reconstructed, the supermaxilla bones include two hundred and sixty-one cases with the range from 12 to 40 gestaional weeks that were normal supermaxilla proved by 2D ultrasound exam, three cases that were the specimens of fetal normal supermaxilla, and eight cases that were abnormal supermaxilla. The normal supermaxilla's imaging success rates of different gestational ages were contrasted. RESULTS: The success rate of normal fetal supermaxilla bone's formation and structure with the 3D image was 97.9% during the gestation of 12~15(+6) weeks, 96.0% of 16~21(+6) weeks, 98.4% of 22~27(+6) weeks, 68.6% of 28~35(+6) weeks, 27.5% of 36~40 weeks. Through the X(2) test, there was no significant difference in the success rate of displaying among the gestation of 12~15(+6) weeks, 16~21(+6) weeks and 22~27(+6) weeks. The success rate during the gestation of 36~40 weeks was the lowest among all the gestation. Big anatomic structures of fetal supermaxilla in 3D images can be shown, but detail cannot. The success rate of cleft palate with 3D image was 100% (8 cases). CONCLUSIONS: 3D ultrasound can supply more detailed and comprehensive information of fetal supermaxilla bone. The better fit examine weeks for obtaining 3D images are within 12~35(+6) weeks, the best fit examine weeks are within 16~27(+6) weeks. The function of 3D skeletal rendering mode image can display cleft palate clearly.

Primary massage using one-finger twining manipulation for treatment of infantile muscular torticollis.

OBJECTIVES: The study objectives were to observe the clinical efficacy of primary massage of twining manipulation with one finger (PMTMOF) versus conventional tuina manipulation for treating muscular torticollis. METHODS: A randomized, controlled, single-blind method was employed. Patients received either PMTMOF (experimental group, n = 265) or conventional tuina manipulation (control group, n = 235), once a day, 20 minutes each, for 15 days of treatment. After four treatment courses, sternocleidomastoid muscle morphology and size were detected using two-dimensional ultrasonography; sternocleidomastoid muscle blood flow was monitored by color Doppler ultrasonography; and head-neck deflection and range of motion were used to determine total curative effects. RESULTS: In the treatment group, 55 patients were cured, 120 patients remarkably responded to the treatment, 75 patients effectively responded, and 15 patients were found to have no response. The total effective response rate to the treatment is 94.34%. In the control group, 15 patients were cured, along with very effective results in 60 patients, effective results in 125 patients, and ineffective results in 35 patients, and the total effective rate is 85.11%. A significant difference in total effective rate was found between experimental and control groups (p < 0.05). CONCLUSIONS: PMTMOF produced more obvious curative treatment effects in infantile muscular torticollis than conventional tuina manipulation and could effectively shorten treatment time and avoid sequelae due to delayed healing.