[Experimental study of dopamine ameliorating the inflammatory damage of olfactory bulb in mice with allergic rhinitis].

Objective: To investigate the effects of dopamine on olfactory function and inflammatory injury of olfactory bulb in mice with allergic rhinitis (AR). Methods: AR mouse model was established by using ovalbumin (OVA), and the mice were divided into two groups: olfactory dysfunction (OD) group and without OD group through buried food pellet test (BFPT). The OD mice were randomly divided into 2 groups, and OVA combined with dopamine (3, 6, 9 and 12 days, respectively) or OVA combined with an equal amount of PBS (the same treatment time) was administered nasally. The olfactory function of mice was evaluated by BFPT. The number of eosinophils and goblet cells in the nasal mucosa were detected by HE and PAS staining. Western blotting, immunohistochemistry or immunofluorescence were used to detect the expression of olfactory marker protein (OMP) in olfactory epithelium, the important rate-limiting enzyme tyrosine hydroxylase (TH) of dopamine, and the marker proteins glial fibrillary acidic protein (GFAP) and CD11b of glial cell in the olfactory bulb. TUNEL staining was used to detect the damage of the olfactory bulb. SPSS 26.0 software was used for statistical analysis. Results: AR mice with OD had AR pathological characteristics. Compared with AR mice without OD, the expression of OMP in olfactory epithelium of AR mice with OD was reduced (F=26.09, P<0.05), the expression of GFAP and CD11b in the olfactory bulb was increased (F value was 38.95 and 71.71, respectively, both P<0.05), and the expression of TH in the olfactory bulb was decreased (F=77.00, P<0.05). Nasal administration of dopamine could shorten the time of food globule detection in mice to a certain extent, down-regulate the expression of GFAP and CD11b in the olfactory bulb (F value was 6.55 and 46.11, respectively, both P<0.05), and reduce the number of apoptotic cells in the olfactory bulb (F=25.64, P<0.05). But dopamine had no significant effect on the number of eosinophils and goblet cells in nasal mucosa (F value was 36.26 and 19.38, respectively, both P>0.05), and had no significant effect on the expression of OMP in the olfactory epithelium (F=55.27, P>0.05). Conclusion: Dopamine can improve olfactory function in mice with AR to a certain extent, possibly because of inhibiting the activation of glial cells in olfactory bulb and reducing the apoptotic injury of olfactory bulb cells.

[A case of spontaneous remission of acute myeloid leukemia with MLL-AF9 rearrangement and abnormal liver function].

Objective: To explore the clinical features and possible pathogenesis of spontaneous remission of acute myeloid leukemia (AML) . Methods: We retrospectively analyzed the clinical data of a patient with spontaneous remission of AML, MLL-AF9 rearrangement, and abnormal liver function in the First Affiliated Hospital of Zhengzhou University, and the relevant pieces of literature were summarized. Results: The patient experienced lung infection, fever, and liver dysfunction and was treated with anti-infection and blood transfusion. After complete response (CR) , the patient remained in CR with mild, indirect bilirubin elevation at 35 months of follow-up. Additionally, 56 cases of adult AML (non-acute promyelocytic leukemia) were reported in the literature from 1990 to June 2021. The cases were checked by bone marrow aspiration, and our patients were summarized and analyzed. Furthermore, 57 patients, including 37 males and 20 females, with a median age of 51 (20-83) years and a median remission time of five months; 52 patients achieved complete remission. In addition, there were five cases with long-term remission and a chromosomal record, with no recurrence so far, three with normal karyotype and two with t (9;11) (q21;q23) . Conclusion: The spontaneous remission of leukemia is rare and may be related to immunosuppression and genes.

[Gadolinium angiography of inner ear in patients with sudden deafness and its clinical analysis].

Objective: To observe the gadolinium imaging findings of inner ear in patients with sudden deafness and to analyze its clinical features. Methods: From November 2017 to July 2020, 21 patients with sudden deafness in the People's Hospital of Dongsheng District, Ordos City were selected as the research objects, including 14 males and 7 females, aged 36-76 years, with a median age of 50 years. The course of disease was 1-19 days, with an average of 5.5 days. The patients received audiology tests, laboratory examination, and intravenous gadolinium angiography, each of whom was scanned twice by 3D-FLAIR sequence: once before intravenous gadolinium injection, and once again 4.5-6.0 h after intravenous gadolinium injection. The following corresponding clinical treatment was given. The imaging manifestations and clinical features were observed. Results: Among 21 cases of sudden deafness in acute stage, the signal intensity of 11 cases was significantly higher than that of the contralateral ear, and 2 cases had vestibular labyrinthine hydrops. In laboratory examination, only 2 cases of total deafness had increased WBC count and faster erythrocyte sedimentation rate, and the rest had no abnormality. The hearing types of 21 patients with sudden deafness were: total deafness in 8 cases, flat decline in 10 cases, low frequency decline in 1 case, high frequency decline in 2 cases. The total effective rate was 57% (12/21). The hearing types of 11 patients with abnormal gadolinium angiography were total deafness in 5 cases, flat decline in 5 cases and high frequency decline in 1 case. The total effective rate was 64% (7/11). Conclusion: Gadolinium angiography is abnormal in some patients with sudden deafness, and the permeability of blood labyrinth barrier may be increased, which is worthy of further study.

LncRNA PCAT-1 regulated cell proliferation, invasion, migration and apoptosis in colorectal cancer through targeting miR-149-5p.

OBJECTIVE: Emerging studies suggested that lncRNA plays an important role in cell progression of multiple cancers. In CRC, the function of some lncRNAs has been verified to be related to cell proliferation, apoptosis, migration and invasion, providing a crucial theoretical basis for the treatment of colorectal cancer. Due to the complexity of the regulation mechanism of cell growth, the regulation mechanism related to lncRNA still needs to be further studied in CRC. PATIENTS AND METHODS: The qRT-PCR assay was used to carry out the expression of prostate cancer-associated ncRNA transcripts 1 (PCAT-1) and miR-149-5p. The Western blots were used to measure the protein expression of CDK4, Cyclin D1, MMP-2, MMP-9, Bcl-2, Bax and ß-actin. Additionally, flow cytometry and MTT assay were used to assess cell apoptosis and cell proliferation, respectively. Moreover, transwell assay was applied to measure the ability of cells migrated and invasion in CRC. Luciferase reporter assay was performed to detect luciferase activities. RESULTS: In this study, lncRNA PCAT-1 expression was significantly upregulated in CRC cells and tissues. More than that, knockdown of lncRNA PCAT-1 inhibited cell proliferation, migration and invasion while promoted cell apoptosis in CRC cells. Of note, lncRNA PCAT-1 directly targeted miR-149-5p and miR-149-5p expression was significantly downregulated in CRC cells and tissues. Moreover, miR-149-3p reversed the suppressive effects of PCAT-1 on the cell growth of CRC cells. CONCLUSIONS: Our study demonstrated that LncRNA PCAT-1 regulated cell proliferation, invasion, migration and apoptosis in colorectal cancer through targeting miR-149-5p and provided a new regulatory mechanism of CRC.

Twist1/2 activates MMP2 expression via binding to its promoter in colorectal cancer.

OBJECTIVE: This study aimed to characterize the effect of Twist2 on epithelial-to-mesenchymal transition (EMT) and the invasive potential of colorectal cancer (CRC) cells and to explore the mechanisms underlying the regulative effect of Twist1 and Twist2 on matrix metalloproteinase 2 (MMP2) expression in CRC. PATIENTS AND METHODS: Data mining was performed in colorectal cancer cohort (COADREAD) in the Cancer Genome Atlas (TCGA-COADREAD). CRC LoVo and HCT116 cells were used as in vitro cell models. RESULTS: CRC tumors with lymphatic invasion (N = 102) had a significantly higher expression of TWIST1 (p = 0.01) and TWIST2 (p = 0.02) than the lymphatic invasion negative cases (N = 228). TWIST2 overexpression enhanced EMT and the invasive potential of the CRC LoVo and HCT116 cells, while TWIST2 knockdown reversed the EMT process and weakened the invasive potential of the cells. TWIST1 and TWIST2 were co-upregulated with MMP2 and MMP9 in COADREAD cohort. TWIST1 or TWIST2 overexpression significantly elevated nuclear ß-catenin accumulation, which is a known signaling pathway elevating MMP2 and MMP9 expression. More importantly, we found that both Twist1 and Twist2 could transcriptionally activate MMP2 via directly binding to its promoter. However, this mechanism was not observed in the MMP9 promoter. CONCLUSIONS: TWIST1/2 is associated with lymphatic invasion in CRC. TWIST2 upregulation enhances EMT and the invasive potential of CRC cells. TWIST1/2 can enhance the Wnt/ß-catenin signaling pathway in CRC cells. In addition, Twist1/2 can bind to the MMP2 promoter and promote its transcription.

Glycolysis gatekeeper PDK1 reprograms breast cancer stem cells under hypoxia.

This corrects the article DOI: 10.1038/onc.2017.368.

Glycolysis gatekeeper PDK1 reprograms breast cancer stem cells under hypoxia.

Glycolysis is critical for cancer stem cell reprogramming; however, the underlying regulatory mechanisms remain elusive. Here, we show that pyruvate dehydrogenase kinase 1 (PDK1) is enriched in breast cancer stem cells (BCSCs), whereas depletion of PDK1 remarkably diminishes ALDH+ subpopulations, decreases stemness-related transcriptional factor expression, and inhibits sphere-formation ability and tumor growth. Conversely, high levels of PDK1 enhance BCSC properties and are correlated with poor overall survival. In mouse xenograft tumor, PDK1 is accumulated in hypoxic regions and activates glycolysis to promote stem-like traits. Moreover, through screening hypoxia-related long non-coding RNAs (lncRNAs) in PDK1-positive tissue, we find that lncRNA H19 is responsible for glycolysis and BCSC maintenance. Furthermore, H19 knockdown decreases PDK1 expression in hypoxia, and ablation of PDK1 counteracts H19-mediated glycolysis and self-renewal ability in vitro and in vivo. Accordingly, H19 and PDK1 expression exhibits strong correlations in primary breast carcinomas. H19 acting as a competitive endogenous RNA sequesters miRNA let-7 to release Hypoxia-inducible factor 1α, leading to an increase in PDK1 expression. Lastly, aspirin markedly attenuates glycolysis and cancer stem-like characteristics by suppressing both H19 and PDK1. Thus, these novel findings demonstrate that the glycolysis gatekeeper PDK1 has a critical role in BCSC reprogramming and provides a potential therapeutic strategy for breast malignancy.

Icariin induces S-phase arrest and apoptosis in medulloblastoma cells.

Medulloblastoma is the most common type of malignant brain tumor in children. Despite a relatively high long-term survival rate, complications still represent great burden for the majority of patients receiving traditional therapy. Therefore, the development of new effective treatments and drugs is urgently needed. A cell counting kit-8 (CCK-8) and colony formation assay were used to evaluate medulloblastoma cell proliferation and colony formation, respectively. Cell cycles and apoptosis were assessed by flow cytometry. A western blot was performed to determine the levels of protein expression. Axenograft model of medulloblastoma was established to evaluate the in vivo anticancer effects of icariin. The CCK-8 assay showed that icariin decreased cell viability in a dose- and time-dependent manner. The colony formation assay indicated that icariin potently inhibited the colony formation ability of Daoy and D341 cells. Icariin-induced proliferation inhibition may be due to S-phase arrest in medulloblastoma cells. In addition, icariin induced apoptosis in a dose-dependent manner, as shown by the results of annexin V/propidium iodide (PI) double staining and Hoechst 33342 staining. Icariin progressively inhibited tumor growth and induced apoptosis in a mouse model. Moreover, cell cycle regulators Cyclin A, CDK2, and Cyclin B1, and apoptosis-related proteins caspase-3, caspase-9, poly (ADP-ribose) polymerase (PARP), and Bcl-2 were modulated in response to treatment with icariin in vitro and in vivo. Our results suggest that icariin may exert anticancer effects. Thus, it is a promising drug for medulloblastoma treatment.

Kinases and phosphatases in ischaemic preconditioning: a re-evaluation.

Activation of several protein kinases occurs during myocardial ischaemia and during subsequent reperfusion. In contrast to the intensive investigation into the significance of kinase activation in cardioprotection, relatively little is known about the role of the phosphatases in this regard. The aim of this study was to re-evaluate the putative roles of PP1 and PP2A in ischaemia/reperfusion and in triggering ischaemic preconditioning. Isolated perfused working rat hearts were subjected to sustained global (15 or 20 min) or regional ischaemia (35 min), followed by reperfusion. Hearts were preconditioned using global ischaemia (1 x 5 or 3 x 5 min, alternated with 5 min reperfusion). To inhibit both PP1 and PP2A cantharidin (5 muM) was used. To inhibit PP2A only, okadaic acid (7.5 nM) was used. The drugs were administered during the preconditioning protocol, before onset of sustained ischaemia (pretreatment) or during reperfusion. Endpoints were mechanical recovery during reperfusion, infarct size and activation of PKB/Akt, p38 MAPK and ERK p42/p44, as determined by Western blot. Pretreatment of hearts with okadaic acid or cantharidin caused a significant reduction in mechanical recovery after 15 or 20 min global ischaemia. Administration of the drugs during an ischaemic preconditioning protocol abolished functional recovery during reperfusion and significantly increased infarct size. Administration of the drugs during reperfusion had no deleterious effects and increased functional recovery in 3 x PC hearts. To find an explanation for the differential effects of the inhibitors depending on the time of administration, hearts were freeze-clamped at different time points during the perfusion protocol. Administration of cantharidin before 5 min ischaemia activated all kinases. Subsequent reperfusion for 5 min without the drug maintained activation of the kinases until the onset of sustained ischaemia. Cantharidin given during preconditioning was associated with activation of p38MAPK and PKB/Akt during reperfusion after sustained ischaemia. However, administration of the drug during reperfusion only after sustained ischaemia caused activation of both PKB/Akt and ERK p42/p44. Phosphatase inhibition immediately prior to the onset of sustained ischaemia or during preconditioning abolishes protection during reperfusion, while inhibition of these enzymes during reperfusion either had no effect or enhanced the cardioprotective effects of preconditioning. It is proposed that inhibition of phosphatases during reperfusion may prolong the period of RISK activation and hence protect the heart.

Supratentorial Brain AVM Embolization with Onyx-18 and Post-Embolization Management. A Single-Center Experience.

SUMMARY: Onyx is increasingly used in endovascular therapy of intracranial arteriovenous malformations (AVMs). However, the embolic effect and post-embolization management are still under discussion. We report our experience in the treatment of supratentorial brain arteriovenous malformations (SBAVMs) with Onyx and discuss post-embolic management. From June 2006 to July 2008, 20 patients with SBAVM were embolized with Onyx. There were 14 men and six women ranging from 14 to 64 years of age (mean 38.3 years). Initial symptoms included spontaneous hemorrhage (n=12), headaches (n=4), seizure (n=3) and incidentally disclosed after head trauma (n=1). After the endovascular procedure, all had subsequent treatment (follow-up angiogram, stereotactic radiosurgery or microsurgery) according to the obliteration degree. At angiography, seven patients (35%, 7/20) were completely obliterated (over 95% closure) after embolization while one suffered a small subarachnoid hemorrhage without permanent clinical sequelae. Four patients (20%, 4/20) were subtotally obliterated (over 80% closure), one patient who suffered severe cerebral edema after embolization underwent decompressive craniectomy, two patients had additional radiosurgery and one patient had follow-up angiogram. Nine patients (45%, 9/20) were partially obliterated (20-80% closure), five patients had additional surgery, two patients had additional radiosurgery and two patients had follow-up angiogram (one patient had intraventricular hemorrhage three months after embolization). Of all 20 AVMs, an average of 2.2 ml Onyx was used per patient and average volume reduction was 80% (range, 30%-99%). Onyx is suitable for embolization of SBAVMs because of its diffuse controllable properties. We suggest clinical follow-up after complete obliteration, additional radiosurgery or angiographic follow-up after subtotal obliteration and additional surgery after partially obliteration. More cases with long-term follow-up are needed to evaluate the long-term prognosis of our post-embolization management.

Analysis of the induction and deinduction of tyrosine aminotransferase in enucleated HTC cells.

Anucleate HTC cells have been used to determine the importance of the nucleus in the regulation of the intracellular levels of tyrosine aminotransferase (TAT) in hepatoma tissue culture (HTC) cells. In the absence of the nucleus, neither the induction of the enzyme by dexamethasone nor its deinduction upon removal of the hormone occurs. Degradation of the enzyme takes place when protein synthesis is inhibited in anucleates by cycloheximide. Therefore, the maintenance of induced levels of enzyme activity after dexamethasone withdrawal from pre-induced anucleates suggest that the nucleus is required for the inactivation of the TAT mRNA.

Synthesis of viral DNA in the cytoplasm of duck embryo fibroblasts and in enucleated cells after infection by avian sarcoma virus.

TWO LINES OF EVIDENCE INDICATE THAT SYNTHESIS OF VIRAL DNA OCCURS IN THE CYTOPLASM OF DUCK EMBRYO FIBROBLASTS INFECTED WITH AVIAN SARCOMA VIRUS: (i) viral DNA is detected first in the cytoplasmic fraction of infected cells and subsequently in the nuclear fraction; and (ii) viral DNA is synthesized at a normal rate in cells infected after enucleation with cytochalasin B. The presence of viral DNA in the cytoplasmic fraction is not a consequence of leakage of newly synthesized viral DNA from the nucleus, since it remains in nuclear fractions late after infection when integration of viral DNA into the host genome is inhibited by ethidium bromide.