Role of the anterior insular cortex microcircuits in observation fear of mice / 中国药理学与毒理学杂志

OBJECTIVE Fear can be learned indi-rectly,but excessive transmission of fear is essential for the development of mental illness.Previous research has indicated that the anterior insular cortex(AIC)may play a crucial role in the process of fear transmission,and abnormal AIC activity is a possible mechanism under-lying various affective disorders.Inhibitory neurons are crucial for maintaining local microcircuit homeostasis.With the support of novel specific neuroregulatory tech-niques,it is now possible to monitor and regulate differ-ent types of neurons in real-time.Therefore,investigating distinct subtypes of inhibitory neurons in the AIC that are involved in fear contagion may provide valuable insights into potential mechanisms underlying mental disorders.METHODS We established a modified observational fear(OF)model.A demonstrator(DM)mouse was placed in an acrylic cup at the center of the apparatus,and two observer(OB)mice were allowed to explore the DM mouse simultaneously from separate areas on either side.During the OF training,electric foot shocks were administered to the DM mouse and freezing,the side and corner time,and social interaction behavior were scored.Next,we characterized the activity patterns of distinct neuronal subtypes in the AIC using GCaMP-based calcium recording.Finally,we employed a Cre-dependent optogenetic approach to selectively modulate excitatory or inhibitory neurons in the AIC,and investigat-ed empathic fear behavior across different Cre transgenic mouse lines(CK2-Cre,PV-Cre,SOM-Cre,VIP-Cre).RESULTS During the training phase,the OB mice exhib-ited significantly higher levels of fear compared to the control group(which did not observe a traumatic event),as evidenced by increased freezing time,decreased interaction time,and increased corner zone time.Calcium fiber recording results suggested that CK2 neurons are involved in risk prediction,while PV and VIP neurons exert inhibitory control on this behavior.Optogenetic silencing of CK2-positive neurons in the AIC through injection of AAV-DIO-NpHR-mCherry in mice demon-strated a significant reduction in empathic fear.Similarly,activation of PV or VIP inhibitory neurons expressing ChR2-eYFP also resulted in a similar effect.However,activation of SOM neurons led to a significant increase in empathic fear.CONCLUSION Our study demonstrated that VIP and PV neuron activity in the AIC attenuates empathetic fear,while SOM and CK2 neuron activity enhances fear expression.These findings shed light on the distinct contributions of various inhibitory interneu-rons in the AIC to fear contagion,indicating their mutual interaction for maintaining local microcircuit homeostasis that regulates empathetic fear behaviors.

A new mouse model to study the empathic fear and pain behavior / 中国药理学与毒理学杂志

OBJECTIVE Human beings possess the ability to indirectly acquire the emotions of others.This also known as emotional contagion or empathy,enables us to rapidly perceive the emotions of others.However,an excessive empathy may result in heightened fear and sensitivity to pain.Therefore,the establishment of appropri-ate animal models for analyzing neural mechanisms underlying empathy would contribute to pharmacological research on pain sensitivity caused by psychological sus-ceptibility.METHODS We used the observed fear para-digm for assessing contagion of negative emotions in mice.To minimize the impact of emotional contagion dif-ferences caused by the subject change,we established a bilateral observation area and the two mice were trained to observe fear simultaneously.First,two observer(OB)mice were placed on either side of the observational area.Next,a demonstrator(DM)mouse was introduced into the cylindrical shock cage located at the center of the apparatus.The shock cage is made of transparent organic plastic with air holes and has provided ample space for free movement by the DM mouse.During the shock stage,DM mice were subjected to electric stimulation while the behaviors of OB mice on both sides was observed,including freezing,the side and corner time,social interaction behavior.Additionally,c-Fos staining was utilized to confirm distinct local brain activities.RESULTS In the habituation stage,OB mice on both sides showed more social preference for DM mouse,as evidenced by an increase in duration time in the designat-ed interaction zone.During the shock phase,OB mice observed the DM mouse receiving electric shocks and displayed significantly higher levels of fear contagion;however,their fear behavior was not entirely consistent.Some mice exhibited a significant increase in freezing time,while others demonstrated a significant increase in corner and side exploration time.We utilized Z-normal-ization to evaluate changes in emotionality across vari-ous behaviors and identified mice with distinct susceptibil-ities.Fos-positive neurons exhibited higher expression levels in susceptible OB mice,primarily concentrated within brain regions associated with the ascending path-ways of pain perception,such as thalamus,the anterior insular cortex,and anterior cingulate cortex.CONCLU-SION In this study,we have developed an innovative experimental facility that integrates various behavioral tests to evaluate empathic behavior in mice.Our findings highlight the robustness of emotionality measures obtained from individual mice by combining this experi-mental model with the Z-scoring method,facilitating screening for empathic fear or pain-susceptible mice and will helpful for pharmacological evaluation.

Poly(ADP-ribose)polymerase inhibitors for ovarian cancer treatment:research advances / 国际药学研究杂志

Ovarian cancer is the leading cause of death in women suffering from cancer,with a high mortality rate in gyneco?logical cancer. Poly(ADP-ribose)polymerase(PARP)inhibitors cause targeted tumor cell death in homologous recombination(HR)-deficient cancers,including breast cancer susceptibility gene(BRCA)tumors,and the mechanism is calledsynthetic lethality. At present,there are three PARP inhibitors approved by FDA for the treatment of advanced ovarian cancer with BRCA-mutation. This pa?per reviews the role of PARP inhibitors in the treatment of ovarian cancer in clinical trial,elaborates the therapeutic mechanism of PARP inhibitors,and lights the way for the development of anti-ovarian cancer drugs.

An ion-pair reversed-phase HPLC method for analyzing the ATP, ADP and AMP contents in hippocampus of mice / 国际药学研究杂志

Objective To establish an ion-pair reverse-phased high-performance liquid chromatography(RP-HPLC)method for simultaneous determination of ATP,ADP and AMP in the hippocampus of mice. Methods The protein of mouse hippocampus was precipitated with perchloric acid,and neutralized with potassium carbonate-methanol mixture. Mobile phase was as follows:50 mmol/L phosphate buffer(buffer for K2HPO4-KH2PO4,pH 6.60,containing 22%methanol,and 4 mmol/L tetrabutylammonium bisul?fate). Shimadzu HPLC system and Agilent C18 column(4.6 mm×250 mm,5μm)filled with the same material pre-column(12.5 mm× 4.6 mm,5μm)were used. The contents of ATP,ADP and AMP in mouse hippocampus were analyzed at a wavelength of 254 nm,the flow rate of 0.6 ml/min and room column temperature. Results Stability tests showed that intra-day and inter-day precision of the method were 1.27%-3.42%and 0.88%-3.52%,respectively,and recovery rates were 95.67%-104.05%. Conclusion The HPLC method established in this study is simple,accurate and efficient in detecting the levels of ATP,ADP,and AMP in mice hippocampus.

Effects of diterpene ginkgolides meglumine injection on neurotransmitters in cerebral ischemia-reperfusion injury rats / 中国药理学通报

Aim To investigate the effects of diterpene ginkgolides meglumine injection (DGMI ) on amino acids and monoamine neurotransmitters in rats with cerebral ischemia/reperfusion injury.Methods In-traluminal suture was applied to establish middle cere-bral artery occlusion (MCAO/R)model with ischemia for 1.5 h and reperfusion for 24 h.After the adminis-tration of DGMI (i.v.),the levels of amino acid and monoamine neurotransmitters in brain tissue were de-tected through HPLC-ECD.Results DGMI down-reg-ulated the concentrations of aspartic acid, glutamic acid,glycine and γ-aminobutyric acid which were in-creased in MCAO/R group.DGMI also reduced the levels of norepinephrine epinephrine,glyoxylic acid, serotonin and 5-HIAA in cortex and hippocampus,and increased adrenaline content compared to the model group.Conclusion DGMI exhibits a protective role in rats with cerebral ischemia /reperfusion injury through regulating amino acids and monoamine neuro-transmitters.

Medical countermeasures against chemical threats:a review of antidotes / 中国药理学与毒理学杂志

Chemical warfare agents and chemical terrorism agents have been identified as one of the major threats to human survival and national security currently. The key to dealing with these threats is the effective medical countermeasures of which specific antidotes take center stage. In the past decade,real or potential chemical threats which has sparked regional conflicts,terrorist activities or chemical accidents intentionally or unintentionally have increased the investment in antidotes research and development worldwide. Here,we introduced the research status on medical countermeasures against chemical threat by giving an overview of the United States ″Countermeasures Against Chemi?cal Threats(CounterACT)Program″,and then the recent research progress in antidotes against nerve agents,sulfur mustard and cyanide toxicities were reviewed.

Techniques of molecular network analysis in study on network pharmacology / 国际药学研究杂志

The technology of network analysis,based on a variety of disciplines theory,is the core technology in the research of network pharmacology,through which we can mine specified information from molecular networks from multi-angle and multi-level. It has been used to investigate and obtain valuable information about the ingredients/drugs with specific pharmacological effects ,the key nodes,modules and motifs with specific biological function,and physiological mechanism of drug action,pathogenesis of dis?ease,or biomarker of disease,especially for the complex disease represented by Alzheimer′s disease(AD). In this paper,the general techniques of network analysis in network pharmacology study are reviewed.

Exploration of the experimental conditions of drug affinity responsive target stability / 国际药学研究杂志

Objective To explore the optimal proteolysis condition for the new technology of target discovery ,drug affinity responsive target stability(DARTS). Methods First,in order to determine the suitable pronase concentration range for DARTS,the extraction of human acute T lymphoblastic leukemia cells(Jurkat)were digested with a range of pronase concentrations(the mass ratio of pronase to protein was 1∶100 to 1∶10000)at room temperature and detected by SDS-PAGE and Coomassie brilliant blue stain?ing. On this basis,in order to obtain the optimal proteolysis condition,we performed DARTS onα-ketoglutarate in combination with SDS-PAGE and gel staining,and observed the effect of different concentrations of pronase(1∶500-1∶5000)and different proteolysis time(5-30 min)on DARTS. The feasibility of the optimum condition was verified by using it on a target known small molecule ,myco?phenolic acid. Results When the protein extraction of Jurkat was hydrolyzed by a range of pronase concentrations,it was entirely hy?drolysed by pronase 1∶100 while showing no significant effect under the condition of pronase 1∶10000. The effect of pronase 1∶500 to 1∶5000 on protein mixture was milder. And approximately 30%-60%of the protein was digested. The protein bands which were protected byα-ketoglutarate could be observed apparently under the conditon of pronase 1∶1000 when the proteolysis time was about 15 min. Then we performed DARTS on mycophenolic acid utilizing this condition(pronase 1∶1000,hydrolysed for 15 min)and obtained sev?eral visible protected protein bands between(4-7)×104. Western blotting results showed that the target protein of mycophenolic acid , IMPDH1,was contained in the protected protein bands. Conclusion The optimal proteolysis condition for DARTS on protein mix?ture throughα-ketoglutarate is obtained.

Insulin signaling pathway and glucose metabolism in Alzheimer′s diseases / 国际药学研究杂志

Alzheimer′s disease(AD)is a degenerative metabolic disease,whose exact pathological mechanism still remains unknown. Currently,studies have found that patients in AD accompany with insulin signaling pathway impairment and cerebral glu?cose metabolism dysfunction. As insulin signaling pathway and cerebral glucose metabolism homeostasis play a key role in AD ,some researches consider AD as“typeⅢdiabetes”. This review aims to discuss the alteration of cerebral insulin signaling pathway and glu?cose metabolism in AD,as well as their relationship with AD. We will also elaborate the advance in anti-AD drugs based on cerebral insulin signaling pathway.

Contribution of luteinizing hormone to Alzheimer′s disease / 国际药学研究杂志

Luteinizing hormone(LH)is a gonadotropin of hypothalamic-pituitary-gonadal axis(HPG),secreted by the anteri?or pituitary. The secretion of LH is directly controlled by the release of gonadotropin releasing hormone(GnRH),acts at the ovaries and testes to stimulate the production of gonadal hormones. Aging leads to increases in LH,and higher serum levels of LH has been ob?served in Alzheimer′s disease(AD)patients when compared to age-matched controls. Evidences from basic research and epidemiologi?cal investigation support the critical role of elevated LH in pathogenic process of AD and deteriorating cognitive decline. Here we sum?marize the recent discoveries containing human AD epidemiological evidence for LH,cognitive impairments resulting from LH activi?ty,LH in AD pathology and LH receptor signaling mechanisms.

Techniques for molecular network construction in network pharmacology study / 国际药学研究杂志

Network construction technology is the basis for network pharmacology research. Data originated from experiment, database and computational prediction can be used to construct drugome network,diseaseome network and molecular interactome net?work. The molecular network construction for the complex diseases represented by Alzheimer′s disease(AD)could be used to identify the important targets,pathogenesis and drug action mechanism,and guide drug development and drug reposition. This paper reviews techniques for molecular network construction in network pharmacology study.

Biological characteristics ofβ-secretase:research advances / 国际药学研究杂志

β-secretase, a typeⅠtransmembrane aspartic protease, initiates β-amyloid protein(Aβ) formation by cleaving β-amyloid precursor protein between Met596 and Asp597 or between Tyr606 and Glu607 to generate N-terminal fragment of Aβ. β-Secretase has many members,and aspartic proteinaseβ-site amyloid precursor protein-cleaving enzyme 1 (BACE1) is the most famous of them. Because BACE1 plays a significant role in the pathogenesis of Alzheimer′s disease (AD) and a neurodegenerative disease, it has been studied fully. In this article, we review the gene and protein structure, transcription and translation, intracellular trafficking and metabolism of BACE1 and summarize substrates and homologous enzymes of BACE1. This work provides some reference for BACE1 as a drug target on AD.

Effect of clioquinol on aggregation ofβ-amyloid in vitro / 国际药学研究杂志

Objective To study whether the metal chelator clioquinol (CQ) can affectβ-amyloid (Aβ) aggregation directly in vitro and whether this effect could be influenced by Zn2+. Methods In the study thioflavin T (Th-T) fluorescence was used to detect the aggregated Aβ. To eliminate the possible false positive results, the absorption spectrum (300 nm to 600 nm) of CQ was scanned, and a competitive binding assay was applied to determine whether Th-T and CQ had the same binding site on Aβ. Circular dichroism spectroscopy was used to detect β-sheet formation of Aβ. Results CQ could decrease the fluorescence intensity, when incubated with monomer Aβor aggregated Aβfor 24 h. Absorption spectra indicated that CQ had no specific absorption peak at 450 nm and 485 nm. Competitive binding assay showed that CQ and Th-T did not bind the same site on Aβ. CD spectra showed that CQ could decrease theβ-sheet formation of Aβ. When incubated with monomer Aβ, CQ decreased the fluorescence intensity in a dose dependent manner, and the IC50 were 6.1μmol/L (without Zn2+) and 4.3μmol/L (with Zn2+);When incubated with aggregated Aβ, CQ decreased the fluorescence intensity in a dose dependent mannerand, and the IC50 was 7.5μmol/L (without Zn2+) and 6.1μmol/L (with Zn2+). Conclusion CQ can inhibit the aggregation of monomer Aβand depolymerize the aggregated Aβdirectly in vitro. Zn2+has little influence on the effect of CQ on Aβ.

Advances in biomarkers of oxidative stress under sulfur mustard exposure / 军事医学

Sulfur mustard [bis (2-chloroethyl) sulfide, sulfur mustard, SM] is considered a powerful chemical warfare agent.There is still no specific antidote due to its complex toxicological mechanism .An intimate knowledge of the toxic mechanisms and pathophysiological changes is important to the treatment of sulfur mustard injury .Oxidative stress is one of the most important pathophysiological processes involved in the toxic effect of sulfur mustard .The study of oxidative stress biomarkers induced by sulfur mustard can help to reveal the role of oxidative stress in sulfur mustard intoxication , which may contribute to better understanding of the toxic mechanism and development of therapeutic measures after sulfur mustard exposure.The research advances in oxidative stress biomarkers in sulfur mustard intoxication are reviewed .

Roles of adenosine receptors in Alzheimer's disease / 药学学报

As an important neurotransmitter, adenosine displays its functions by acting on the adenosine receptors. Recent studies have shown that the distribution, expression and balance among subtypes of adenosine receptors are closely related with cognitive activities, and changes of adenosine receptors play key roles in neurodegenerative disorders including Alzheimer's disease. It has been pointed out that prolonged activation of adenosine receptors by high level adenosine may lead to the disturbance of balance among adenosine receptor subtypes. This imbalance mainly performed as increased expression of A2a receptor and decreased expression of A1 receptor, and enhancement of the excitatory signals mediated by A2a receptor and weakened inhibitory signals mediated by A1 receptor. Changes of these two subtypes of adenosine receptors may lead to a lot of disorders of neurological activities which developed into dysfunction of cognition to the end. These findings imply that the potential of maintaining the balance among adenosine receptors on the treatment of AD would facilitate both the revealing of the mechanism and the cure of AD.

Progress in technology for glycomics / 中国药理学与毒理学杂志

Technologies for glycomics usually involve methods for separation and purification of poly-saccharides, and separation, structure resolution, quantification, property investigation and function comment of glycan chains. Because of the different biochemical properties of glycoproteins, proteogly-cans and glycolipids, the separation and purification of polysaccharides involve corresponding fractional precipitation, boric acid affinity, titanium dioxide, affinity chromatography, size exclusion method, and gel filtration chromatography column chromatography methods. The lectins, water affinity chromatogra-phy , solid phase extraction and other technologies could be applied to the oil enrichment of high pure and specific glycan chains. The structure of glycan chains can be analyzed using lectin microarray technolo-gy, mass spectrometry, and derivatization markers of glycan chains. lsotope labelling and metabolic labeling can be used to quantify glycan chains. The glycan biological function can be better understood using glycan chain structure analysis software and database of glycan chains by bioinformatics.

Establishment of the metabolic syndrome rodent model / 中国药理学通报

AimThe aim of this study was to establish a rodent model with similar characters of human metabolic syndrome(MS).Methods Three species mice and Wistar rats were fed with high energy chows(HEC)for 6 to 23 weeks.Animals were weighted every week.Fasting blood glucose(FBG)together with total cholesterol(TC)and low density lipoprotein-cholesterol(LDL-C)were investigated by oxidase test every two week.And fasting blood insulin(FINS)was determined by radioimmunoassay.Homeostasis model assessment of insulin resistance(HOMA-IR)was calculated as FBG×FINS/22.5.At the end of the experiment,oral glucose tolerance test(OGTT)was performed.Then animals were decapitated,and coel-fat and orchio-fat were collected and weighted to calculate the visceral fat coefficient(VFC).Results FBG,serum TC and LDL-C significantly increased(P<0.01)after 6 weeks feed of HEC in KM mice.The mice also formed abdominal obesity and insulin resistant together with impairment of glucose tolerance(P<0.05 or P<0.01).Though similar to the KM mice,C57BL/6 and BALB/c mice couldn't form abdominal obesity while the latter had increased body weight(P<0.05 or P<0.01).Wistar rats formed hyperlipidemia from 1 to 10 week and hyperglycemia from 10 to 23 week together with insulin resistance and impaired glucose tolerance(P<0.05 or P<0.01).Conclusion KM mice feed with HEC for 6 weeks could successfully establish metabolic syndrome mice model which might be suitable for drug-screening,the major characters includes the formation of abdominal obesity(increase of VFC),the increase of serum TC,LDL-C,FBG and HOMA-IR,and the decrease of OGTT.

Carotid arterial wave intensity in assessing the hemodynamic change in patients with chronic heart failure / 生物医学工程学杂志

Wave intensity (WI) is a newly proposed hemodynamic index that assesses the working condition of cardiovascular system. The purpose of this study is to investigate the clinical value of WI in assessing the function of cardiovascular system in patients with chronic heart failure (CHF). We used a combined Doppler and echo-tracking system to calculate the carotid artery WI in 16 patients with CHF and 35 normal subjects, and examined the characteristics of WI. The main WI indices included W1, W2, NA, R-W1 and W1-W2. Compared with those in the control group, W1 and W2 were significantly decreased in CHF group (5.44 +/- 3.24 vs. 10.05 +/- 46.33, P < 0.01, 0.83 +/- 0.42 vs. 1.50 +/- 0.95, P < 0.01, respectively), the temporal indices R-W1 shortened (141.06 +/- 41.14 vs. 109.77 +/- 22.50, P < 0.05) and W1-W2 prolonged (214.63 +/- 39.93 vs. 260.51 +/- 20.58, P < 0.01), and the (R-W1)/(W1-W2) ratio was significantly larger in CHF group (0.69 +/- 0.26 vs. 0.43 +/- 0.10, P < 0.01). WI provides a great deal of information about the dynamic behavior of the interaction between the heart and vascular system, so it is a new quantitative approach for analysis of CHF.

Research progress in the relationship of autocrine motility factor,autocrine motility factor receptor and the nervous system / 中国药理学通报

Autocrine motility factor (AMF) plays an important role in the stimulation of the migration and motility of cells, especially the generation, migration and angiogenesis of tumor. Recently, it has been found that AMF has three isoforms, ATX-t, ATX-m and PD-I alpha. The PD-I alpha isoform is specifically expressed in the brain, which plays extensive functions in nervous system, such as regulating neural development and differentiation, promoting neurotrauma repair, inducing neuropathic pain, even contributing neurodegeneration under some circumstances. This indicates the close relationship of AMF/AMFR and the pathophysiology of the nervous system. This paper mainly reviews the function of AMF and AMFR and its possible mechanism in the nervous system.

The effects of Liuwei Dihuang decoction on the differential expression genes in the hippocampus of senescence-accelerated mouse / 中国药理学通报

Aim To disclose the molecular mechanism of Liuwei Dihuang decoction(LW)enhances the cognitive function of central nervous system, the effects of Liuwei Dihuang decoction on the differential expression genes in the hippocampus of senescence-accelerated mouse was studied. Methods There were 8 gene expression patterns, such as SAMP8 and SAMR1, SAMP8 as negative control and SAMR1 as positive control, huperzine A-treated SAMP8 and SAMP8 as negative control, LW-treated SAMP8 and SAMP8 as negative control, were compared and assessed by use of the differential expression cDNA microarray of the hippocampus of SAMP8 and SAMR1. The response genes of LW were compared. Results LW had significant modulating effects on some of the gene expressions. Expressions of genes, such as DUSP12, NSF, STUB1, CaMKⅡ?, AMFR, UQCRFS1 and other 11 novel genes without any functional clues changed significantly. These genes involved in the protein-tyrosine phosphatase family, the AAA(ATPases associated with diverse cellular activities)gene family, the serine/threonine protein kinases family, ubiquitin ligase, mitochondrial function and so on. Conclusions These results suggested LW effects on the cognitive impairments might be multi-mechanism and these genes might be the potential gene targets for LW effects on the impairments.