Parkin regulates IGF2BP3 through ubiquitination in the tumourigenesis of cervical cancer.

BACKGROUND: Insulin-like growth Factor 2 mRNA-binding protein 3 (IGF2BP3) is a highly conserved RNA-binding protein and plays a critical role in regulating posttranscriptional modifications. METHODS: Immunoprecipitation was used to examine the interaction of Parkin and IGF2BP3. Mass spectrometry was performed to identify the ubiquitination sites of IGF2BP3. RNA-immunoprecipitation was conducted to examine the target genes of IGF2BP3. Xenograft mouse model was constructed to determine the tumorigenesis of IGF2BP3. RESULTS: IGF2BP3 expression is negatively correlated with Parkin expression in human cervical cancer cells and tissues. Parkin directly interacts with IGF2BP3, and overexpression of Parkin causes the proteasomal degradation of IGF2BP3, while knockdown of PARK2 increases the protein levels of IGF2BP3. Mechanistically, in vivo and in vitro ubiquitination assays demonstrated that Parkin is able to ubiquitinate IGF2BP3. Moreover, the ubiquitination site of IGF2BP3 was identified at K213 in the first KH domain of IGF2BP3. IGF2BP3 mutation results in the loss of its oncogenic function as an m6A reader, resulting in the inactivation of the phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signalling pathways. In addition, IGF2BP3 mutation results in the attenuation of Parkin-mediated mitophagy, indicating its inverse role in regulating Parkin. Consequently, the tumourigenesis of cervical cancer is also inhibited by IGF2BP3 mutation. CONCLUSION: IGF2BP3 is ubiquitinated and regulated by the E3 ubiquitin ligase Parkin in human cervical cancer and ubiquitination modification plays an important role in modulating IGF2BP3 function. Thus, understanding the role of IGF2BP3 in tumourigenesis could provide new insights into cervical cancer therapy.

Within-leaf chloroplasts and nitrogen allocation to thylakoids in relation to photosynthesis during grain filling in maize.

Nitrogen (N) is an important contributor to photosynthetic rate (Pn). However, during grain-filling stage in maize, some leaf N is remobilized to meet demands for grain protein accumulation rather than photosynthetic demands. Therefore, plants that can maintain a relatively high Pn during the N remobilization process would have the key to achieving both high grain yields (HGY) and high grain protein concentrations (HGPC). In this study, we investigated two high-yielding maize hybrids in photosynthetic apparatus and N allocation in a two-year field experiment. During grain filling, XY335 had a higher Pn and photosynthetic N-use efficiency than ZD958 had in the upper leaf, but not in the middle or lower leaves. In the upper leaf, the diameter and area of the bundle sheath (BS) were larger and the distance between bundle sheaths was greater in XY335 than in ZD958. XY335 had more bundle sheath cells (BSCs) and a larger BSC area, as well as a larger chloroplast area in the BSC, resulting in a higher total number and total area of chloroplasts in the BS. XY335 also had higher stomatal conductance (gs), intercellular CO2 concentration, and N allocation to the thylakoids. No genotypic differences were found in mesophyll cell ultrastructure, N content and starch content in the three types of leaves. Therefore, a trifecta of higher gs, greater N allocation to thylakoids for photo-phosphorylation and electron transport, and more and larger chloroplasts promoting CO2 assimilation in the BS confers a high Pn to simultaneously achieve HGY and high HGPC in maize.

Imatinib and methazolamide ameliorate COVID-19-induced metabolic complications via elevating ACE2 enzymatic activity and inhibiting viral entry.

Coronavirus disease 2019 (COVID-19) represents a systemic disease that may cause severe metabolic complications in multiple tissues including liver, kidney, and cardiovascular system. However, the underlying mechanisms and optimal treatment remain elusive. Our study shows that impairment of ACE2 pathway is a key factor linking virus infection to its secondary metabolic sequelae. By using structure-based high-throughput virtual screening and connectivity map database, followed with experimental validations, we identify imatinib, methazolamide, and harpagoside as direct enzymatic activators of ACE2. Imatinib and methazolamide remarkably improve metabolic perturbations in vivo in an ACE2-dependent manner under the insulin-resistant state and SARS-CoV-2-infected state. Moreover, viral entry is directly inhibited by these three compounds due to allosteric inhibition of ACE2 binding to spike protein on SARS-CoV-2. Taken together, our study shows that enzymatic activation of ACE2 via imatinib, methazolamide, or harpagoside may be a conceptually new strategy to treat metabolic sequelae of COVID-19.

Agmatine attenuates the discriminative stimulus and hyperthermic effects of methamphetamine in male rats.

Methamphetamine abuse remains an alarming public heath challenge, with no approved pharmacotherapies available. Agmatine is a naturally occurring cationic polyamine that has previously been shown to attenuate the rewarding and psychomotor-sensitizing effects of methamphetamine. This study examined the effects of agmatine on the discriminative stimulus and hyperthermic effects of methamphetamine. Adult male rats were trained to discriminate 0.32 mg/kg methamphetamine from saline. Methamphetamine dose dependently increased drug-associated lever responding. The nonselective dopamine receptor antagonist haloperidol (0.1 mg/kg) significantly attenuated the discriminative stimulus effects of methamphetamine (5.9-fold rightward shift). Agmatine (10-100 mg/kg) did not substitute for methamphetamine, but significantly attenuated the stimulus effects of methamphetamine, leading to a maximum of a 3.5-fold rightward shift. Acute 10 mg/kg methamphetamine increased the rectal temperature by a maximum of 1.96±0.17°C. Agmatine (10-32 mg/kg) pretreatment significantly attenuated the hyperthermic effect of methamphetamine. Agmatine (10 mg/kg) also significantly reversed methamphetamine-induced temperature increase. Together, these results support further exploration of the value that agmatine may have for the treatment of methamphetamine abuse and overdose.

Interactions between imidazoline I2 receptor ligands and acetaminophen in adult male rats: antinociception and schedule-controlled responding.

RATIONALE: Recent evidence suggests that imidazoline I2 receptor ligands are suitable for combination therapy with opioids. Quantitative analysis of I2 receptor ligands combined with non-opioid drugs is necessary for the justification of alternative pain therapies. OBJECTIVE: This study systematically examined the antihyperalgesic and response rate-suppressing effects of selective I2 receptor ligands (2-BFI and phenyzoline) alone and in combination with acetaminophen. METHODS: Von Frey and Hargreaves tests were used to examine the antihyperalgesic effects of drugs in complete Freund's adjuvant (CFA)-induced inflammatory pain in rats. Food-reinforced schedule-controlled responding was used to assess the rate-suppressing effects of study drugs. Dose-addition and isobolographic analyses were used to assess drug-drug interactions for all assays. RESULTS: 2-BFI (3.2-17.8 mg/kg, i.p.), phenyzoline (17.8-100 mg/kg, i.p.), and acetaminophen (56-178 mg/kg, i.p.) all dose-dependently produced significant antinociceptive effects. When studied as combinations, 2-BFI and acetaminophen produced infra-additive to additive interactions while phenyzoline and acetaminophen produced additive to supra-additive interactions. The same drug combinations suppressed response rate in a supra-additive manner. CONCLUSIONS: Quantitative analysis of the antihyperalgesic and response rate-suppressing effects suggests that I2 receptor ligands are not well suited to combination therapy with acetaminophen.

A rare case of malignant pediatric ectomesenchymoma arising from the cerebrum.

Malignant ectomesenchymoma is a rare tumor that contains both ectodermal and mesenchymal elements. So far, only 7 patients with a manifestation in the cerebrum (with confirmed clinicopathological data) have been reported. A 4-year-old girl was present at our hospital with a 3-week history of intermittent sudden dizzy with no apparent cause. MRI showed an irregular enhanced lesion in the left frontal-parietal lobe and lateral ventricle with peripheral gadolinium-enhancement with a significant surrounding edema. Total removal of the tumor was performed. Histological examination of the resected tumor revealed a mixed astrocytoma and anaplastic ependymoma component with undifferentiated mesenchymal spindle cell component. Generally speaking, the main malignant part in most cases of malignant ectomesenchymoma (MEM) is the mesenchymal component. In the present case, the malignant component was both in the mesenchymal and ectodermal part. In particular, the mesenchymal part was mainly composed of spindle cells, and the ectodermal part primarily consisted of gliomatous component and anaplastic ependymoma component. The patient was then treated with chemotherapy and as regard to the prognosis, there was no evidence of tumor recurrence at the 5 months' follow-up. The long term follow-up is still in progress.

Reduced tumourigenicity of EG7 after RANTES gene transfer and the underlying mechanism.

INTRODUCTION: Chemokine ligand 5, also known as CCL5 or regulated on activation normal T-cell expressed and secreted (RANTES), is a chemokine expressed in inflamed tissue and capable of inducing migration of immature dendritic cells (DCs) or Langerhans cells. In this study, we explored the effect of RANTES on EG7 cells. MATERIAL AND METHODS: In vivo, RANTES gene transfer reduced the tumourigenic capacity of EG7 and prolonged the survival of tumour-bearing mice. To reveal the underlying mechanism, we performed the following experiments and provided evidence to support our hypothesis of RANTES gene therapy for EG7. Higher natural killer (NK) cell and cytotoxic T lymphocyte (CTL) activity was induced after RANTES gene transfer, accompanied by higher levels of Th1 type cytokines (IL-2 and IFN-γ). RESULTS: Tumour necrosis was also markedly observed in the tumour tissues after RANTES gene transfer, which was attributed to reduced expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP-2). CONCLUSIONS: We draw the conclusion that reduced tumourigenicity of EG7 after RANTES gene transfer can be attributed to higher NK cell and CTL activity, anti-angiogenesis and higher levels of Th1 type cytokines induced by RANTES. These results support the notion that higher chemokine expression in tumour tissue elicits potent anti-tumour immunity.