Sonogenetics is preferred for neuroregulation and the treatment of brain diseases due to its noninvasive properties. Ultrasonic stimulation produces thermal and mechanical effects, among others. Since transient receptor potential vanilloid 1 (TRPV1) could be activated at 42 °C, it is overexpressed in the M1 region of the mouse motor cortex to sense the change of temperature upon being stimulated by focused ultrasound. Whether the heat generated by ultrasonic stimulation could activate TRPV1 in the M1 region and induce changes in electromyography (EMG) signals collected from the mice\'s triceps was carefully verified. The position of the focused ultrasound and the temperature of the tissue at the location of the focused position were simulated using COMSOL software and verified via experiments. For Neuro-2a cells with TRPV1 overexpression, 42 °C could activate the TRPV1 and induce calcium influx. For mice with TRPV1 overexpression in the M1 region, tissue temperature of >42 °C in the M1 region induces an increased number of cfos, suggesting that neurons with overexpressed TRPV1 in the M1 region can be activated using focused ultrasound. Furthermore, when the temperature is >42 °C, the peak-to-peak value of the EMG signal for mice with TRPV1 overexpression in the M1 region was higher than that for mice without TRPV1 overexpression. The immunohistochemical results showed that ultrasound was not harmful to the stimulation site. The noninvasive ultrasound stimulation combined with thermosensitive protein TRPV1 overexpressed in neurocytes as sonothermogenetics technology has great potential to be used for the treatment of neurological diseases.

Endometritis is an inflammatory disease of the endometrium, which is responsible for endometrial dysfunction, decidualization failure, and increased incidence of early pregnancy loss. SCM-198, a synthetic form of leonurine, is well known to possess anti-inflammatory effects. SCM-198 has been reported to display beneficial effects on endometritis. However, the specific mechanisms of SCM-198 in preventing endometritis remain unknown. In this study, we focused on the molecular mechanism of SCM-198 in inhibiting endometritis. The anti-inflammatory effects and the related signaling pathways of SCM-198 were studied in vitro using human endometrial stromal cells (hESCs). Reverse transcriptase-polymerase chain reaction and western blot analysis results demonstrated that SCM-198 markedly inhibited lipopolysaccharide (LPS)-induced endometrial inflammatory response by suppressing the LPS-JNK-cJUN/cFOS-TLR4-NF-κB pathway. The preventive and therapeutic effects of SCM-198 on endometrial inflammation were explored by using a mouse model of LPS-induced endometritis. SCM-198 produced essentially the same effects when administered either post-treatment (after LPS) or pre-treatment (before LPS) via vaginal or intraperitoneal administration. In vivo results indicated that SCM-198 is a potential effective drug for the treatment of endometritis.

It is unclear how social drinking can contribute to the development of addiction in susceptible individuals. However, alcohol\'s aversive properties are a well-known factor contributing to its abuse. The lateral habenula (LHb) is a key brain structure responding to various aversive stimuli, including those related to alcohol. We recently reported that ethanol at 10 mM or less that can be achieved by social drinking activates many LHb neurons and drives aversive conditioning. The current study sought to identify LHb circuits that are activated by a low-dose of ethanol using immunohistochemistry and anatomic tracing techniques on adult Sprague-Dawley rats. We showed here that an intraperitoneal injection of ethanol (0.25 g/kg), resulting in a blood ethanol concentration of 5.6 mM, significantly increased the number of cFos immunoreactive (IR) neurons in the LHb. Most of the ethanol-activated cFos-IR LHb neurons expressed vGluT2 (vesicular glutamate transporters 2, a marker of a glutamatergic phenotype). These LHb neurons projected to the ventral tegmental area (VTA), rostromedial tegmental nucleus (RMTg), and dorsal raphe. Moreover, injections of the anterograde tracer AAV-CaMKIIa-eGFP into the lateral hypothalamus produced a significant amount of labeled fibers with vGluT2 positive terminals on the ethanol-activated LHb cells. These results indicate that the LHb neurons stimulated by a low-dose of ethanol project to the VTA, RMTg, and dorsal raphe, and receive excitatory projections from the lateral hypothalamus. These neurocircuits may play a crucial role in mediating the initial aversive effects produced by a low-dose of ethanol.

Previously, we reported that fluoxetine acts on 5-HT2B receptor and induces epidermal growth factor receptor (EGFR) transactivation in astrocytes. Recently, we have found that chronic treatment with fluoxetine regulates Caveolin-1 (Cav-1)/PTEN/PI3K/AKT/glycogen synthase kinase 3β (GSK-3β) signaling pathway and glycogen content in primary cultures of astrocytes with bi-phasic concentration dependence. At low concentrations fluoxetine down-regulates Cav-1 gene expression, decreases membrane content of PTEN, increases PI3K activity and increases phosphorylation of GSK-3β and increases its activity; at high concentrations fluoxetine acts on PTEN/PI3K/AKT/GSK-3β in an inverse fashion. Here, we present the data indicating that acute treatment with fluoxetine at lower concentrations down-regulates c-Fos gene expression via PI3K/AKT signaling pathway; in contrast at higher concentrations fluoxetine up-regulates c-Fos gene expression via MAPK/extracellular-regulated kinase (ERK) signaling pathway. However, acute treatment with fluoxetine has no effect on Cav-1 protein content. Similarly, chronic effects of fluoxetine on Cav-1 gene expression are suppressed by inhibitor of PI3K at lower concentrations, but by inhibitor of MAPK at higher concentrations, indicating that the mechanism underlying bi-phasic regulation of Cav-1 gene expression by fluoxetine is opposing effects of PI3K/AKT and MAPK/ERK signal pathways on c-Fos gene expression. The effects of fluoxetine on Cav-1 gene expression at both lower and higher concentrations are abolished by AG1478, an inhibitor of EGFR, indicating the involvement of 5-HT2B receptor induced EGFR transactivation as we reported previously. However, PP1, an inhibitor of Src only abolished the effect by lower concentrations, suggesting the relevance of Src with PI3K/AKT signal pathway during activation of EGFR.

Bone marrow-derived mesenchymal stem cells (BMSCs) are proposed as the cells of origin of several subtypes of osteosarcoma (OS). However, signals that direct BMSCs to form different subtypes of OS are unclear. Here we show that the default tumor type from spontaneously transformed p53 knockout (p53_KO) BMSCs is osteoblastic OS. The development of this default tumor type caused by p53 loss can be overridden by various oncogenic signals: RAS reprograms p53_KO BMSCs into undifferentiated sarcoma, AKT enhances osteoblastic OS, while cFOS promotes chondroblastic OS formation. We focus on studying the mechanism of cFOS-induced chondroblastic OS formation. Integrated genome-wide studies reveal a regulatory mechanism whereby cFOS binds to the promoter of a key chondroblastic transcription factor, Sox9, and induces its transcription in BMSCs. Importantly, SOX9 mediates cFOS-induced cartilage formation in chondroblastic OS. In summary, oncogenes determine tumor types derived from BMSCs, and the cFOS-SOX9 axis is critical for chondroblastic OS formation.

BACKGROUND: Andrographolide (Andro) is reported to inhibit hepatoma tumor growth in our previous studies. This study aims to further search the critical signals involved in such Andro-induced inhibition.
METHODS: The anti-tumor effect of Andro was evaluated in vivo. Cancer PathwayFinder RT(2) Profiler™ PCR array was used to find the altered genes. Real-time PCR was used to detect the mRNA expression. Protein expression was detected by Western-blot analysis, enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining. Activator protein-1 (AP-1) transcriptional activity was detected by luciferase reporter assay.
RESULTS: Andro (10mg/kg) inhibited hepatoma tumor growth in vivo. The expression of four genes decreased in Andro-treated tumor tissues. Among which, vascular endothelial growth factor (VEGFD) was the highest decreased gene. The decreased VEGFD expression was further confirmed by real-time PCR and immunohistochemical staining assay. Andro decreased VEGFD mRNA and protein expression in hepatoma Hep3B and HepG2 cells. Andro also decreased VEGFD amount in Hep3B cell supernatant. Andro decreased cFos protein expression and its translocation into nucleus, and also reduced AP-1 luciferase activity. Further results showed that Andro induced polyubiquitination of cFos. Proteasome inhibitor MG132 reversed the decreased expression of cFos protein, and the decreased mRNA and protein expression of VEGFD. SP600125, an inhibitor of c-Jun N-terminal kinase (JNK), reversed the decreased expression of cFos and VEGFD induced by Andro.
CONCLUSIONS: Andro decreased VEGFD expression in hepatoma cancer cells via inducing c-fos protein degradation, which will contribute to its anti-cancer activity, and JNK plays some roles in regulating this process.