The central amygdala (CeA) is a crucial modulator of emotional, behavioral, and autonomic functions, including cardiovascular responses. Despite its importance, the specific circuit by which the CeA modulates blood pressure remains insufficiently explored. Our investigations demonstrate that photostimulation of GABAergic neurons in the centromedial amygdala (CeM^GABA), as opposed to those in the centrolateral amygdala (CeL), produces a depressor response in both anesthetized and freely-moving mice. In addition, activation of CeM^GABA axonal terminals projecting to the nucleus tractus solitarius (NTS) significantly reduces blood pressure. These CeM^GABA neurons form synaptic connections with NTS neurons, allowing for the modulation of cardiovascular responses by influencing the caudal or rostral ventrolateral medulla. Furthermore, CeM^GABA neurons targeting the NTS receive dense inputs from the CeL. Consequently, stimulation of CeM^GABA neurons elicits hypotension through the CeM-NTS circuit, offering deeper insights into the cardiovascular responses associated with emotions and behaviors.
Animals ; GABAergic Neurons/physiology* ; Male ; Central Amygdaloid Nucleus/physiopathology* ; Hypotension/physiopathology* ; Mice ; Blood Pressure/physiology* ; Mice, Inbred C57BL ; Solitary Nucleus/physiology* ; Photic Stimulation ; Neural Pathways/physiology*

Objective:To discuss the effect of microRNA(miR)-30c-5p on the proliferation,migration,and invasion of the human prostate cancer cells(LNCap),and to clarify its possible mechanism.Methods:The LNCap cells were divided into LNCap group(without plasmid transfection),miR-30c-5p mimic group(transfected with miR-30c-5p mimic),mimic NC group(transfected with miR-30c-5p mimic NC),sh-DNA damage inducible transcript 4(DDIT4)group(transfected with sh-DDIT4),sh-NC group(transfected with sh-DDIT4 NC),miR-30c-5p mimic+pc-DNA3.1-NC group(co-transfected with miR-30c-5p mimic and pc-DNA3.1 empty vector),and miR-30c-5p mimic+pc-DNA3.1-DDIT4 group(co-transfected with miR-30c-5p mimic and pc-DNA3.1-DDIT4 over-expression plasmid).The RWPE-1 cells were cultured normally.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-30c-5p and DDIT4 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of DDIT4 protein in the cells in various groups;CCK-8 method was used to detect the proliferation rates of the LNCap cells in various groups;Transwell assay was used to detect the numbers of the invasion LNCap cells in various groups;Scratch assay was used to detect the scratch healing rates of LNCap cells in various groups;dual-luciferase reporter assay was used to detect the targeting relationship between miR-30c-5p and DDIT4.In the in vivo tumor formation experiment,18 male BALB/c nude mice were divided randomly into blank group,agomiR-NC group(transfected with agomiR-30c-5p NC),and agomiR-30c-5p group(transfected with agomiR-30c-5p);there were six mice in each group.The mice in agomiR-NC group and agomiR-30c-5p group were subcutaneously injected with LNCap cells,while the mice in blank group were given an equal volume of physiological saline.The volumes of tumor of the mice in various groups were detected.HE staining was used to observe the morphology of prostate cancer tissue the mice of in various groups;RT-qPCR method and immunofluorescence staining were used to detect the expression levels of miR-30c-5p and DDIT4 mRNA and the fluorescence intensities of DDIT4 protein in prostate cancer tissue of the mice in various groups.Results:The In vitro prostate cancer cell experiment results showed that compared with RWPE-1 cells,the expression level of miR-30c-5p in the prostate cancer LNCap cells was decreased(P<0.01),and the expression levels of DDIT4 mRNA and protein were increased(P<0.05 or P<0.01).After 48 of transfection,compared with LNCap group and mimic NC group,the expression level of miR-30c-5p in the LNCap cells in miR-30c-5p mimic group was increased(P<0.01).Compared with LNCap group and sh-NC group,the expression level of DDIT4 mRNA in the LNCap cells in sh-DDIT4 group was decreased(P<0.01).Compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of miR-30c-5p in The LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 mRNA in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 protein in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.05).The CCK-8 method results showed that compared with LNCap group and mimic NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the proliferation rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The Transwell assay results showed that compared with LNCap group and mimic NC group,the number of the invasion LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the number of invasion LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the number of the invasion LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The scratch assay results showed that compared with LNCap group and mimic NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the scratch healing rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The dual-luciferase reporter assay results showed that compared with the LNCap cells co-transfected with WT-DDIT4 and mimic NC,the luciferase activity of the LNCap cells co-transfected with WT-DDIT4 and miR-30c-5p mimic was decreased(P<0.01).The in vivo nude mouse tumor formation experiment results showed that on the 3 rd,6 th,9 th,12 th,and 15th days after cell injection,compared with blank group and agomiR-NC group,the tumor volumes of the nude mice in agomiR-30c-5p group were decreased(P<0.05).The HE staining results showed that in prostate cancer tissue of the mice in blank group and agomiR-NC group,the cell nuclei were enlarged,and nucleoli were prominent and deformed.In the mice in agomiR-30c-5p group,some regions of prostate cancer tissues results showed neatly arranged cells with normally shaped nuclei.The RT-qPCR and immunofluorescence staining showed that compared with agomiR-NC group,the expression level of miR-30c-5p in prostate cancer tissue of the mice in agomiR-30c-5p group was increased(P<0.01).Compared with blank group and agomiR-NC group,the expression level of DDIT4 mRNA in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).DDIT4 protein was mainly expressed in the cytoplasm.Compared with blank group and agomiR-NC group,the fluorescence intensity of DDIT4 protein in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).Conclusion:The expression level of miR-30c-5p in the prostate cancer LNCap cells is decreased,and it inhibits the proliferation,migration,and invasion of the prostate cancer cells by targeting downregulation of DDIT4,thereby participating in the occurrence and development of prostate cancer.

Adrenocortical carcinoma(ACC)is the most prevalent malignant tumor of the adrenal gland and is characterized by a poor pro-gnosis in its advanced stages.Surgical resection of the tumors is typically limited to patients diagnosed in the clinical stages Ⅰ and Ⅱ,lead-ing to a high postoperative recurrence rate.The combination of mitotane(M)with etoposide(E),doxorubicin(D),and cisplatin(P)(EDP-M)is currently the only approved first-line treatment regimen for advanced ACC.However,the efficacy of chemotherapy and radiation therapy in ACC remains limited.If the EDP-M regimen proves ineffective,there are no standardized or universally accepted second-line systemic treat-ment alternatives.Research advancements in the molecular mechanisms of ACC in recent years has led to increasing investigations on tyr-osine kinase inhibitors(TKIs)targeting EGFR,VEGFR,and mTOR,as well as immune checkpoint inhibitors(ICIs).Moreover,previous studies have identified mutations in CTNBB1,TP53,KDM5A,CENP-H,and other genes that may serve as therapeutic targets or biomarkers,thereby expanding the treatment options available for ACC.ICIs are effective against diverse cancer types,including non-small cell lung cancer(NSCLC),liver cancer,renal cell cancer,and urothelial cancer.Ongoing exploration into targeted therapies and immunotherapy,especially combination treatments,holds the promise of extending the survival of patients and enhancing their quality of life.

Objective To investigate the changes of dorsal root ganglia(DRG)neurons in rats with voltage-gated sodium channel 1.7 loss-of-function(Nav1.7 LOF)and its involvement in congenital insensitivity to pain.Methods Immunofluorescence,Western blot,and alternative splicing analysis were used to verify the expression deletion of DRG neurons and Nav1.7 in spinal cord of Nav1.7 LOF rats.Using antibodies for peripherin,calcitonin gene-related peptide(CGRP),isolectin B4(IB4),and neuronal nuclear antigen(NeuN)in the DRG and spinal cord tissues,immunofluorescence staining was performed to observe the differences in DRG neuronal sub-populations and central terminal projections between Nav1.7 LOF rats and wild-type rats.Bulk RNA-Seq and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the difference of gene expression in the DRG neurons between Nav1.7 LOF and WT rats.Following 55℃ thermal stimulation of the hind paw in Nav1.7 LOF rats,c-Fos and phospho-extracellular signal-regulated kinase(p-ERK)antibody staining was used to observe the activation of DRG neurons and spinal dorsal horn neurons.Results Nav1.7 was absent in the DRG neurons and spinal cord of Nav1.7 LOF rats.Compared with wild-type rats,Nav1.7 LOF rats exhibited a decreased proportion of IB4+non-peptidergic neurons in DRG tissues.Additionally,there was a significant loss of central projections of IB4+neurons in the spinal dorsal horn.The RNA-Seq and RT-qPCR results demonstrate that in Nav1.7 LOF rats,the gene expression of Mas-related G protein-coupled receptor(MRGPR),markers for 1B4+non-peptidergic neurons,was downregulat-ed.Compared with WT rats,there was no difference in c-Fos and p-ERK staining in DRG neurons of Nav1.7 LOF rats following ther-mal stimulation of the hindpaw.However,there was no c-Fos or p-ERK staining in spinal dorsal horn neurons,suggesting that the neu-rons were not activated.Conclusion Nav1.7 LOF rats exhibit a decreased proportion of IB4+non-peptideric neurons in DRG tissue and their central terminal projections.Noxious stimuli could activate DRG neurons,but could not activate dorsal horn neurons,suggesting the pain signal is interrupted between the central terminals of DRG neurons and the spinal dorsal horn.

Objective To investigate the changes of dorsal root ganglia(DRG)neurons in rats with voltage-gated sodium channel 1.7 loss-of-function(Nav1.7 LOF)and its involvement in congenital insensitivity to pain.Methods Immunofluorescence,Western blot,and alternative splicing analysis were used to verify the expression deletion of DRG neurons and Nav1.7 in spinal cord of Nav1.7 LOF rats.Using antibodies for peripherin,calcitonin gene-related peptide(CGRP),isolectin B4(IB4),and neuronal nuclear antigen(NeuN)in the DRG and spinal cord tissues,immunofluorescence staining was performed to observe the differences in DRG neuronal sub-populations and central terminal projections between Nav1.7 LOF rats and wild-type rats.Bulk RNA-Seq and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the difference of gene expression in the DRG neurons between Nav1.7 LOF and WT rats.Following 55℃ thermal stimulation of the hind paw in Nav1.7 LOF rats,c-Fos and phospho-extracellular signal-regulated kinase(p-ERK)antibody staining was used to observe the activation of DRG neurons and spinal dorsal horn neurons.Results Nav1.7 was absent in the DRG neurons and spinal cord of Nav1.7 LOF rats.Compared with wild-type rats,Nav1.7 LOF rats exhibited a decreased proportion of IB4+non-peptidergic neurons in DRG tissues.Additionally,there was a significant loss of central projections of IB4+neurons in the spinal dorsal horn.The RNA-Seq and RT-qPCR results demonstrate that in Nav1.7 LOF rats,the gene expression of Mas-related G protein-coupled receptor(MRGPR),markers for 1B4+non-peptidergic neurons,was downregulat-ed.Compared with WT rats,there was no difference in c-Fos and p-ERK staining in DRG neurons of Nav1.7 LOF rats following ther-mal stimulation of the hindpaw.However,there was no c-Fos or p-ERK staining in spinal dorsal horn neurons,suggesting that the neu-rons were not activated.Conclusion Nav1.7 LOF rats exhibit a decreased proportion of IB4+non-peptideric neurons in DRG tissue and their central terminal projections.Noxious stimuli could activate DRG neurons,but could not activate dorsal horn neurons,suggesting the pain signal is interrupted between the central terminals of DRG neurons and the spinal dorsal horn.

Carotid atherosclerosis (CAS) is closely associated with the decline of cognitive function in the elderly, which can lead to persistent or progressive cognitive function and neurological dysfunction. Vascular cognitive impairment (VCI) is considered to be an intervenable disease. Studies have shown that CAS is one of the main causes of VCI. Further study on the relationship between CAS and VCI will help to better prevention and treatment of VCI.

Estrogen is an important hormone secreted by the female reproductive system. Its main function is associated with reproduction, growth and development. Studies have shown that estrogen has biological functions such as regulating vasoconstriction, antioxidant stress, anti-inflammatory and neuroprotection, and also affects brain structure and network. Studies have shown that estrogen is closely associated with the occurrence and development of white matter hyperintensities (WMHs). This article reviews the relationship between estrogen and menopausal hormone replacement therapy and WMHs, and their possible pathophysiological mechanisms.

Neutrophil to lymphocyte ratio (NLR) can provide information of local or systemic inflammation and immune status. With the increasing attention to the role of inflammatory and immune factors in vascular cognitive impairment (VCI), it is very important to find new serum inflammatory markers for early identification and intervention of VCI. This article reviews the related research on NLR, VCI and their risk factors, expounds the role of inflammation in the pathogenesis of VCI, and provides help for the diagnosis and prevention of VCI.

Objective To explore the methods of improving diagnosis correctness between the patients with prostate cancer and benign prostatic hyperplasia.Methods Totally 87 patients with benign prostatic hyperplasia or prostate cancer which confirmed by MRI and prostate biopsy for the PSA significantly increased in our hospital from July 2013 to March 2016 were collected.By using the three methods of the PI-RADS V2 score,the T2WI+DWI/ADC+DCE-MRI+MRS and PI-RADS V2 score+MRS to diagnose,and comparing with the pathology results,the diagnostic consistency of the two physicians were analyzed.The sensitivity,accuracy and specificity of three ways were compared,and the correlation between PI-RADS V2 score and Gleason score were calculated.Results The diagnostic consistency of the two physicians:PI-RADS V2 score,K=0.951;T2WI+DWI/ADC+DCE-MRI+MRS score,K=0.838;PI-RADS V2+MRSI score,K=0.937.The correlation between PI-RADS V2 score and Gleason score,r=0.871,P=0.001 4;diagnostic sensitivity,specificity and accuracy of PI-RADS V2 score were 77.3％,74.4％,75.9％;diagnostic sensitivity,specificity and accuracy of T2WI+DWI/ADC+DCE-MRI+MRS were 88.6％,76.7％,82.8％;diagnostic sensitivity,specificity and accuracy of PI-RADS V2+MRSI score were 86.4％,81.4％,83.9％,respectively.Conclusion Compared with the traditional diagnostic methods,the combination of new prostate report and data system and MRSI can improve the diagnostic accuracy of prostate cancer and benign prostatic hyperplasia.The PI-RADS V2 score is more objective and accurate in the description of the lesion,but the low signal of benign hyperplastic nodules in transitional zone should be dialogued carefully through a variety of image parameters.

Objective Using ROC curve to determine the best cut-off point of serum HE-4 in the diagnosis of ovarian cancer and provide important value to diagnosis early ovarian cancer.Methods The levels of serum HE-4 in 68 ovarian cancer pa-tients,42 ovarian benign tumor patients and 30 healthy female were detected by electrochemistry irradiance method.The ROC curve was drawn and the cut-off point of HE-4 was determined by statistical software.Results The levels of serum HE-4 were all non-normal distribution in the groups of ovarian cancer,ovarian benign tumor and healthy controls.Whats more,there was no significant difference between ovarian benign tumor group and normal control group.And compared with benign ovarian tumors and normal control group,the level of HE-4 in ovarian cancers was significantly increased (P <0.01).It would be best for diagnosis when the level of serum HE-4 was 108pmol/L in ovarian cancer.Youden’s index showed the maximum (0.713)and the sensitivity and specificity of diagnosing were 77.9% and 93.1% respectively.The positive predictive value was 91.4% and negative predictive value was 91.4%.At the same time,the positive likelihood ratio was 11.6 and the negative likelihood ratio was 0.2,odds ratio reached to 47.3.Conclusion The detection of HE-4 is an ideal mark for diagnosing and excluding ovarian cancer.Selecting 108 pmol/L as ovarian cancer diagnosis point is relative appro-priate.