Vitexicarpin suppresses malignant progression of colorectal cancer through affecting c-Myc ubiquitination by targeting IMPDH2.

BACKGROUND: Colorectal cancer (CRC) is the second most common cause of cancer-related mortality and is characterised by extensive invasive and metastatic potential. Previous studies have shown that vitexicarpin extracted from the fruits of Vitex rotundifolia can impede tumour progression. However, the molecular mechanisms involved in CRC treatment are still not fully established. PURPOSE: Our study aimed to investigate the anticancer activity, targets, and molecular mechanisms of vitexicarpin in CRC hoping to provide novel therapies for patients with CRC. STUDY DESIGN/METHODS: The impact of vitexicarpin on CRC was assessed through various experiments including MTT, clone formation, EDU, cell cycle, and apoptosis assays, as well as a tumour xenograft model. CETSA, label-free quantitative proteomics, and Biacore were used to identify the vitexicarpin targets. WB, Co-IP, Ubiquitination assay, IF, molecular docking, MST, and cell transfection were used to investigate the mechanism of action of vitexicarpin in CRC cells. Furthermore, we analysed the expression patterns and correlation of target proteins in TCGA and GEPIA datasets and clinical samples. Finally, wound healing, Transwell, tail vein injection model, and tissue section staining were used to demonstrate the antimetastatic effect of vitexicarpin on CRC in vitro and in vivo. RESULTS: Our findings demonstrated that vitexicarpin exhibits anticancer activity by directly binding to inosine monophosphate dehydrogenase 2 (IMPDH2) and that it promotes c-Myc ubiquitination by disrupting the interaction between IMPDH2 and c-Myc, leading to epithelial-mesenchymal transition (EMT) inhibition. Vitexicarpin hinders the migration and invasion of CRC cells by reversing EMT both in vitro and in vivo. Additionally, these results were validated by the overexpression and knockdown of IMPDH2 in CRC cells. CONCLUSION: These results demonstrated that vitexicarpin regulates the interaction between IMPDH2 and c-Myc to inhibit CRC proliferation and metastasis both in vitro and in vivo. These discoveries introduce potential molecular targets for CRC treatment and shed light on new mechanisms for c-Myc regulation in tumours.

Comparison of aqueous and non-aqueous capillary electrophoresis for the determination of four benzalkonium chloride homologues in compound chemical disinfectants.

A new method was established for the simultaneous analysis of four homologous benzalkonium chlorides (dodecyldimethylbenzyl ammonium chloride, tetradecyldimethylbenzyl ammonium chloride, hexadecyldimethylbenzyl ammonium chloride, and octadecyldimethylbenzyl ammonium chloride) in compound chemical disinfectants using non-aqueous capillary electrophoresis (CE) based on a micellar electrokinetic chromatography mode with direct ultraviolet detection. The separation was performed on an uncoated fused quartz capillary with a total length of 60.2 cm and a diameter of 25 µm. The separation buffer consisted of a mixture of methanol/acetonitrile (60:40, v/v) containing 70 mmol/L sodium acetate, 60 mmol/L trifluoroacetic acid and 20 mmol/L sodium dodecyl sulfate. The sample buffer was a methanol solution containing only 2 mmol/L trifluoroacetic acid. The separation voltage was set at 8 kV with a working current of approximately 2.3 µA. The detection wavelength was 214 nm. Under optimal conditions, the limit of detection and limit of quantification for these four benzalkonium chlorides (BACs) were 1.0 mg/L and 5.0 mg/L, respectively. Good linearities were observed in the concentration ranges from 5.0 to 100.0 mg/L, with correlation coefficients above 0.999 for all compounds. The recoveries of these four BACs ranged from 92.5 % to 109.1 % with relative standard deviations below 4.7 %. With the new method, all four BACs could be analyzed in a single injection. In contrast, the aqueous CE method in the National Standard GB/T 26369-2020 only allowed for the simultaneous analysis of the first three homologous. The new method demonstrated the improved peak shape compared to the aqueous CE method and then was successfully applied to the analysis of 19 commercially available samples, such as object table disinfectants, hand sanitizers, and disinfectant wipes, which claimed to contain quaternary ammonium compound. The results obtained using the new method were compared with those of the aqueous CE of the National Standard Method, and no statistically significant differences were observed. The new method is simple in pre-treatment and provides accurate results, making it highly suitable for routine analysis.

[Simultaneous determination of triclosan，triclocarban and p-chloro-m-xylenol in disinfectant，personal care products and oiltment by nonaqueous capillary electrophoresis].

Triclosan （TCS）， triclocarban （TCC）， and p-chloro-m-xylenol （PCMX） are some of the most widely used antibiotics because of their broad-spectrum and highly-efficient bactericidal effects. In the context of disinfection， the National Standard GB 38598-2020 stipulates that the contents of the effective ingredients present in a disinfectant must be specified， wherein their range must fall within 90%-110% of the specified central value. To ensure a suitable product quality， analysis by high performance liquid chromatography （HPLC） is recommended by both the GB/T 27947-2020 and GB/T 34856-2017. However， the results analyzed according to the National Standard method often exceed the declared contents， thereby indicating the necessity to establish a new method based on a completely different principle （e. g.， capillary electrophoresis）， especially since it was not possible to analyze TCS， TCC， and PCMX in a single injection using the National Standard method. Moreover， using this method， large amounts of methanol were consumed， which could be potentially harmful to both operators and the environment. In terms of their water solubilities， this decreases in the order of PCMX>TCS>TCC， wherein TCC is insoluble in water. As such， the use of nonaqueous capillary electrophoresis （NACE） based on running buffer solutions prepared in pure organic solvents （e. g.， methanol or acetonitrile） is necessary. In this paper， a new NACE approach combined with an ultraviolet detection method was developed for the simultaneous analysis of TCS， TCC， and PCMX in disinfectants， personal care products， and ointments. For this purpose， an uncoated fused silica capillary （20 cm×50 µm， total length=30.2 cm） was used as the separation column with a separation buffer composed of 14 mmol/L sodium borate， 2 g/L polyethylene glycol （PEG） 20000， and 0.5 mmol/L dodecyltrimethylammonium bromide （DTAB） in methanol. Following optimization of the separation parameters， the complete and simultaneous separation of TCS， TCC， and PCMX was achieved when the sample solution was prepared using 5 g/L PEG 20000 in methanol-acetonitrile （50∶50， v/v）. It was possible to directly inject the sample into the analysis system after a simple dilution with the sample medium， and no interference was observed in any of the sample electropherograms when a separation voltage and detection wavelength of -12 kV and 214 nm were employed， respectively. Furthermore， TCS， TCC， and PCMX showed good linear relationships with their corrected peak areas within a mass concentration range of 1-100 mg/L， and the correlation coefficients （r） were all greater than 0.99. Moreover， the limits of detection （LODs， S/N=3） and limits of quantification （LOQs， S/N=10） were determined to be 0.2 and 1 mg/L， respectively. The spiked recoveries ranged from 94.5% to 104.4% with relative standard deviations of ≤4.8% in all cases. Subsequently， the established method was used to analyze 31 commercial samples， including hand sanitizer， disinfectant， baby powder， and antibacterial cream. A comparative analysis of HPLC， the developed NACE method， and our previously reported micellar electrokinetic chromatographic （MEKC） method was also carried out for the quantitative determination of TCS， TCC， and PCMX. Although no statistically significant differences were observed among the three methods， the results determined for 16 out of the 31 samples did not match the claimed contents. These results therefore indicate the necessity to further control the compositions of disinfectant products. Our results indicate that the newly established NACE method can be an important alternative to HPLC for routine laboratory analyses， especially considering that it minimizes waste generation， requires only a simple sample pretreatment process， and exhibits a good selectivity to the target compounds. It is therefore hoped that the NACE method will be incorporated into the National Standard method in the near future.

Sexually dimorphic effects of estrogen receptor 2 deletion in the dorsal raphe nucleus on emotional behaviors.

Sex differences in emotional behaviors and affective disorders have been widely noted, of which sexually dimorphic secretion of gonadal steroid hormones such as estrogen is suspected to play a role. However, the underlying neural mechanisms remain poorly understood. We noted that the expression of estrogen receptor 2 (Esr2, or ERß), a key mediator of estrogen signaling in the brain, was enriched in the dorsal raphe nucleus (DRN), a region involved in emotion regulation. To investigate whether DRN Esr2 expression confers sex-specific susceptibility or vulnerability in emotional behaviors, we generated a conditional allele of Esr2 that allowed for site-specific deletion of Esr2 in the DRN via local injection of Cre-expressing viruses. DRN-specific Esr2 deletion mildly increased anxiety behaviors in females, as shown by decreased time spent in the center zone of an open field in knockout females. By contrast, DRN Esr2 deletion had no effects on anxiety levels in males, as demonstrated by knockout males spending comparable time in the center zone of an open field and open arms of an elevated-plus maze. Furthermore, in the tail suspension test, DRN Esr2 deletion reduced immobility, a depression-like behavior, in a male-biased manner. Together, these results reveal sex-specific functions of DRN Esr2 in regulating emotional behaviors and suggest targeted manipulation of DRN Esr2 signaling as a potential therapeutic strategy to treat sex-biased affective disorders.

A circuit from the ventral subiculum to anterior hypothalamic nucleus GABAergic neurons essential for anxiety-like behavioral avoidance.

Behavioral observations suggest a connection between anxiety and predator defense, but the underlying neural mechanisms remain unclear. Here we examine the role of the anterior hypothalamic nucleus (AHN), a node in the predator defense network, in anxiety-like behaviors. By in vivo recordings in male mice, we find that activity of AHN GABAergic (AHNVgat+) neurons shows individually stable increases when animals approach unfamiliar objects in an open field (OF) or when they explore the open-arm of an elevated plus-maze (EPM). Moreover, object-evoked AHN activity overlap with predator cue responses and correlate with the object and open-arm avoidance. Crucially, exploration-triggered optogenetic inhibition of AHNVgat+ neurons reduces object and open-arm avoidance. Furthermore, retrograde viral tracing identifies the ventral subiculum (vSub) of the hippocampal formation as a significant input to AHNVgat+ neurons in driving avoidance behaviors in anxiogenic situations. Thus, convergent activation of AHNVgat+ neurons serves as a shared mechanism between anxiety and predator defense to promote behavioral avoidance.

Thalidomide-based Pt(IV) prodrugs designed to exert synergistic effect of immunomodulation and chemotherapy.

Combination of immune- and chemo-therapy has become a new trend in cancer treatment. Food and Drug Administration (FDA)-approved immune-modulatory agent, thalidomide, can modulate the related proteins of upstream signaling pathway of programmed cell death-ligand 1 (PD-L1), including nuclear transcription factor κB (NF-κB), hypoxia inducible factor-1α (HIF-1α), epidermal growth factor receptor (EGFR), and signal transducer and activator of transcription 3 (STAT3), all acting as key antitumor target proteins. In this work, we conjugated thalidomide with oxidized cisplatin to construct multi-functional Pt(IV) prodrugs, named thaliplatins 4-6, to investigate the anti-tumor effect of immuno- and chemo-therapy. Among them, thaliplatin 6 exerted remarkable cytotoxicity against the tested cancer cell lines, showing 15-26 and 9-20 times higher IC50 values than those of single cisplatin or the combination of cisplatin + thalidomide, respectively. Moreover, thaliplatin 6 could rapidly accumulated into cells, markedly triggered DNA damage, and induced cell S phase arrest and apoptosis, as well as inhibited cell migration and invasion in breast carcinoma cell line (MCF-7). Fluorescent confocal and western blotting experiments proved that 6 significantly regulated NF-κB, EGFR, HIF-1α and phosphor-signal transducer and activator of transcription 3 (p-STAT3), and simultaneously inhibited PD-L1 expression to interrupt programmed cell death 1 (PD-1)/PD-L1 signaling pathway, suggesting a synergistic action of cisplatin and thalidomide. Most strikingly, in vivo tests indicated that 6 effectively decreased tumor growth with no observable systemic toxicity, being superior to the anticancer efficacy of cisplatin.

Automatic Femoral Deformity Analysis Based on the Constrained Local Models and Hough Forest.

Clinically, Taylor spatial frame (TSF) is usually used to correct femoral deformity. The first step in correction is to analyze skeletal deformities and measure the center of rotation of angulation (CORA). Since the above work needs to be done manually, the doctor's workload is heavy. Therefore, an automatic femoral deformity analysis system was proposed. Firstly, the Hough forest and constrained local models were trained on the femur image set. Then, the position and size of the femur in the X-ray image were detected by the trained Hough forest. Furthermore, the position and size were served as the initial values of the trained constrained local models to fit the femoral contour. Finally, the anatomical axis line of the proximal femur and the anatomical axis line of the distal femur could be drawn according to the fitting results. According to these lines, CORA can be found. Compared with manual measurement by doctors, the average error of the hip joint orientation line was 1.7°, the standard deviation was 1.75, the average error of the anatomic axis line of the proximal femur was 2.9°, and the standard deviation was 3.57. The automatic femoral deformity analysis system meets the accuracy requirements of orthopedics and can significantly reduce the workload of doctors.

Targeting ROS-AMPK pathway by multiaction Platinum(IV) prodrugs containing hypolipidemic drug bezafibrate.

Alterations in lipid metabolism, commonly disregarded in the past, have been accepted as a hallmark for cancer. Exploring cancer therapeutics that interrupt the lipid metabolic pathways by monotherapy or combination with conventional chemotherapy or immunotherapy is of great importance. Here we modified cisplatin with an FDA-approved hypolipidemic drug, bezafibrate (BEZ), via the well-established Pt(IV) strategy, affording two multi-functional Pt(IV) anticancer agents cis,cis,trans-[Pt(NH3)2Cl2(BEZ)(OH)] (CB) and cis,cis,trans-[Pt(NH3)2Cl2(BEZ)2] (CP) (BEZ = bezafibrate). The Pt(IV) prodrug CB exhibited an enhanced anticancer activity up to 187-fold greater than the clinical anticancer drug cisplatin. Both CB and CP had less toxicity to normal cells, showing higher efficacies and superior therapeutic indexes than cisplatin. Mechanism studies revealed that the bezafibrate-conjugated Pt(IV) complex CB, as a representative, could massively accumulate in A549 cells and genomic DNA, induce DNA damage, elevate intracellular ROS levels, perturb mitochondrial transmembrane potentials, activate the cellular metabolic sensor AMPK, and result in profound proliferation inhibition and apoptosis. Further cellular data also provided evidence that phosphorylation of AMPK, as a metabolic sensor, could suppress the downstream HMGB1, NF-κB, and VEGFA, which may contribute to the inhibition of angiogenesis and metastasis. Our study suggests that the antitumor action of CB and CP mechanistically distinct from the conventional platinum drugs and that functionalizing platinum-based agents with lipid-modulating agents may represent a novel practical strategy for cancer treatment.

Ketoplatin in triple-negative breast cancer cells MDA-MB-231: High efficacy and low toxicity, and positive impact on inflammatory microenvironment.

Triple-negative breast cancer (TNBC) shares the molecular features facilitating epithelial-to-mesenchymal transition (EMT), which contributed to tumor invasion and metastasis. A platinum(IV) conjugate ketoplatin deriving from FDA-approved drugs cisplatin and ketoprofen was designed and prepared to enhance antitumor activity and suppress EMT in TNBC via positive impact on inflammatory microenvironment by modulating COX-2 signal. As a prodrug, ketoplatin afforded 50.26-fold higher cytotoxicity than cisplatin against TNBC mesenchymal-stem cell-like MDA-MB-231 cells, partly attributing to its dramatic increase of cellular uptake and DNA damage. More importantly, EMT progress in MDA-MB-231 was markedly restrained by ketoplatin, resulting from the suppression of vimentin and N-cadherin mediated by down-regulated COX-2. Further in vivo investigation exhibited that ketoplatin effectively inhibited tumor growth and reduced systemic toxicity compared to cisplatin. Overall, ketoplatin possessed high antitumor activity and low toxicity against TNBC MDA-MB-231 in vitro and in vivo.

Specific Hypothalamic Neurons Required for Sensing Conspecific Male Cues Relevant to Inter-male Aggression.

The hypothalamus regulates innate social interactions, but how hypothalamic neurons transduce sex-related sensory signals emitted by conspecifics to trigger appropriate behaviors remains unclear. Here, we addressed this issue by identifying specific hypothalamic neurons required for sensing conspecific male cues relevant to inter-male aggression. By in vivo recording of neuronal activities in behaving mice, we showed that neurons expressing dopamine transporter (DAT+) in the ventral premammillary nucleus (PMv) of the hypothalamus responded to male urine cues in a vomeronasal organ (VNO)-dependent manner in naive males. Retrograde trans-synaptic tracing further revealed a specific group of neurons in the bed nucleus of the stria terminalis (BNST) that convey male-relevant signals from VNO to PMv. Inhibition of PMvDAT+ neurons abolished the preference for male urine cues and reduced inter-male attacks, while activation of these neurons promoted urine marking and aggression. Thus, PMvDAT+ neurons exemplify a hypothalamic node that transforms sex-related chemo-signals into recognition and behaviors.

[Determination of A2 ß-casein and total ß-casein in cow milk and milk powder by capillary zone electrophoresis].

ß-casein (ß-CN) is one of the major casein proteins in cow milk. There are 13 different variants documented for ß-CN in cow milk, among which A1 and A2 are the major variants. The separation and quantitation of A2 ß-CN are imperative for A2 dairy products. A new capillary zone electrophoresis (CZE) method with UV detection at 214 nm was established for the separation and quantification of the A2 variant and total ß-CN content in cow milk and milk powders. The separation of ß-CN variants was achieved on bare fused silica capillaries (50 µm×30/40 cm (effective/total length)). The separation buffer was a mixture of 4 mol/L urea, 0.2% (mass fraction) hydroxypropyl methylcellulose, 140 mmol/L citric acid, and 50 mmol/L disodium hydrogen phosphate buffer (pH 2.7). The corrected peak areas and the concentrations of total ß-CN and the A2 variant showed good linearity, with correlation coefficients (r2) ranging from 0.9968 to 0.9997. The intra-day precisions for A2 ß-CN and total ß-CN determination in four samples (two pasteurized milk samples and two milk powder samples) were in the ranges of 2.4%-4.7% and 2.6%-4.8%, respectively. The inter-day precisions for A2 ß-CN and total ß-CN determination in four samples were in the ranges of 4.0%-6.3% and 3.9%-6.7%, respectively. The recoveries of A2 and total ß-CN ranged from 85.5% to 106.4%. With the established CZE method, the A2 ß-CN variant and total ß-CN protein in liquid and powder bovine milk products could be separated and accurately quantified. By calculating the A2 ß-CN content in the total ß-CN, the quality of A2 dairy products can be evaluated, and this in turn would aid in the protection of consumer rights.

[Application, development, and challenges of capillary electrophoresis in disease prevention and control].

Since the advent of commercial instruments in 1989, capillary electrophoresis (CE) has advanced considerably, with improvement in reproducibility and accuracy in many application fields. CE is predominantly used in research on disease prevention and control, and hygienic chemical inspection. The applications of CE range from assessment of inorganic anions and cations in drinking water to that of biological macromolecules, such as nucleic acids, in pathogenic microorganisms. Since the analytical capacity of CE ranges from inorganic ions to cell, it has become an indispensable technique in this field, particularly in public health emergency and epidemic management. Universal non-targeted analyses to detect possible pathogens, and the capability of rapid and accurate testing of large numbers of specimens are required. In the analyses of polymerase chain reaction (PCR) products, nucleic acid sequencing, mutation detection and genotyping, food-borne disease pathogens, and vaccine analyses, CE methods characterized by high through-put and sensitivity are necessary. In the public health sector, CE is essential in the analyses of food (including emergency analyses for food poisoning), cosmetics, and disinfectants. Satisfactory results of the FAPAS (Food Analysis Performance Assessment Scheme) and domestic proficiency tests indicated the accuracy of CE in quantitative analyses. Application of CE in disease prevention and control is challenged by a number of new molecular biological methods, as optimizing CE methods may not be feasible, and results are difficult to interpret. CE methods, including transformation of peaks to identification of pathogens, can be an arduous task. Thus, end-users prefer using commercialized CE systems and reagents in their routine work. Alternatively, CE methods for analysis of small molecules in product analyses, such as food safety, cosmetics and disinfectant testing, is commonly performed. A plethora of studies published within the decade, indicate that CE is still an active research area in hygienic chemical inspection. To a large extent, CE has not been used for routine analysis in the centers for disease control and prevention, accredited laboratories in China, nor regulatory agencies worldwide. This may be due to the lack of practical protocols for the standards, and the misconceptions regarding the ease of use of CE, which could have hindered its widespread application. Although CE is an environmental friendly technique with minimal usage of toxic chemicals, few standard methods of CE exist in agriculture, environmental protection, food, beverage, chemical, and pharmaceutical industries in the United States, Britain, Europe, Japan, India, Brazil, Russia, and China. Since 2002, CE was used in our laboratory to analyze a large variety of samples. We found that once the CE method has been fully verified and described in detail, it was easily standardized. It is not necessary to screen the equivalent chromatographic column, or to use a specific liquid chromatographic (LC) column. This can effectively circumvent the challenge of shifting peak orders caused by different LC column selectivity. Once combined with general, high sensitivity detectors, CE can be used in the detection of bacteria or viruses in food safety, and play a greater role in the field of disease prevention and control. In the present review, applications of CE in nucleic acid detection for viruses and bacteria, analysis of vaccines, routine testing on food, dietary supplements, medical foods, cosmetics and disinfectants, proficiency tests, and emergency analyses of food poisoning were summarized. The applications and challenges of CE in the field of disease control and prevention were analyzed, and development of this technique was prospected.

[Non-targeted screening of triazole fungicides in tomatoes by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry].

In order to cope with the differences in international trade regulations, it is necessary to establish a rapid non-targeted screening method for triazole fungicides in food. Therefore, a non-targeted method based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established for the rapid screening of triazole fungicides in tomatoes. Tomato samples were extracted with acetonitrile and then purified by QuEChERS. The purified solution was separated on a BEH C18 column (100 mm×2.1 mm, 1.7 µm) using a gradient mobile phase consisting of acetonitrile and water (both containing 0.1% formic acid). Full information tandem mass spectrometry (MSE) acquisition using electrospray ion source under positive ion mode was performed. The 19 triazole fungicides were divided into eight groups based on their structural differences. The secondary mass fragments of eight typical standards in the eight groups of triazole fungicides were analyzed, and then, the regularities of the characteristic fragment ions and neutral loss fragments produced by collision-induced dissociation of the triazole fungicides were summarized. The fragmentation regularity was further verified by the other 11 triazole fungicides. A total of 16 tomato samples collected from local supermarkets in Beijing were screened using this non-targeted method. Diniconazole was identified and confirmed in one sample, which demonstrated that the method could achieve rapid and accurate non-targeted screening of triazole fungicides in tomatoes. The proposed method is highly efficient, sensitive, and accurate, thus providing a reference for screening triazole fungicide residues in food.

A highly selective colorimetric and fluorescent probe for quantitative detection of Cu2+/Co2+: The unique ON-OFF-ON fluorimetric detection strategy and applications in living cells/zebrafish.

Identifying and detecting similar target cations through combining "turn on" and "turn off" fluorescence mechanism is effective and challenging. Now a new colorimetric and ON-OFF-ON fluorescent probe N'-((7-(diethylamino)-2-oxo-2H-chromen-3-yl)methylene)-3-hydroxy-2-naphthohydrazide (L) was reported, which could detect Cu2+ and Co2+ in phosphate buffered CH3CH2OH-H2O solvent system. With the assistance of glutathione and pH adjustment, a unique ON-OFF-ON fluorescence detection strategy could be achieved for distinguishing Cu2+ and Co2+. The emission of probe could recover from the L-Cu2+ and L-Co2+ system by addition of GSH or adjusting pH value to 4, respectively, which is due to the abolishment of paramagnetic Cu2+/Co2+. Based on fluorescence titration experiments, the limit of detection was determined as 3.84 × 10-9 M and 4.55 × 10-9 M for Cu2+ and Co2+, respectively. Meanwhile, the detection limit reached 6.21 × 10-8 M for Cu2+ and 6.96 × 10-8 M for Co2+ according to absorbance signal output. Fast recognition of Cu2+/Co2+ can be achieved by obvious color changes from green to colorless under UV light, as well as from yellow to orange-red in room light. The binding mode of L toward Cu2+ and Co2+ have been systematically studied by Job's plot analysis, ESI-MS, IR and density functional theory calculations. Most strikingly, further practical applications of the probe L in fluorescence imaging were investigated in MCF-7 cells and zebrafish due to its low cytotoxicity and good optical properties, suggesting that L could serve as a fluorescent sensor for tracking Cu2+ and Co2+in vivo.

[Uniform and orthogonal designs on experimental design and optimization of micellar electrokinetic capillary chromatographic method for the simultaneous analysis of three nucleoside antivirus drugs in disinfectant and anti/bacteriostatic products].

To prevent the abuse of nucleoside-based antiviral drugs in disinfectant and anti/bacteriostatic products, a new micellar electrokinetic chromatographic (MEKC) method for the simultaneous determination of three nucleoside antiviral drugs (ganciclovir, acyclovir and penciclovir) was established. Three factors and seven levels uniform design and four factors and four levels orthogonal design were used to optimize the concentration of sodium dodecyl sulfate (SDS), sodium dihydrogen phosphate (NaH2PO4) and sodium borate (Na2B4O7) in the running buffer. The three nucleoside antiviral drugs were separated from each other in the shortest time to obtain effective separation through uniform and orthogonal designs on experimental design and optimization. The separation was performed on an uncoated fused-silica capillary with 50 µ m i. d. and a total length of 30.2 cm (effective length:20 cm). An optimized buffer solution consisting of 25 mmol/L NaH2PO4, 10 mmol/L Na2B4O7 (pH 7.41) and 140 mmol/L dodecyl sulfate sodium was used for separation. The applied voltage was 10 kV, the injected pressure and time was at 0.003 Pa for 4 s. The wavelength of the detection was 250 nm. Under the optimum conditions, the corrected area and the concentration had good linearity. The correlation coefficients (r) were not less than 0.9995. The limits of detection were all 2.0 mg/kg. The limits of quantitation were all 7.0 mg/kg. The relative recoveries ranging from 85.4% to 104.1% with RSDs all lower than 8.0%. It showed that the method was suitable for the detection of ganciclovir, acyclovir and penciclovir in disinfectant and anti/bacteriostat products with simplicity and rapidity.

A behavioral mechanistic investigation of the role of 5-HT1A receptors in the mediation of rat maternal behavior.

Previous work suggests that 5-HT1A receptors play a special role in rodent maternal aggression, but not in other aspects of maternal care (e.g. pup retrieval and nest building). The present study re-assessed the basic effects of 5-HT1A activation or blockade on various maternal responses in postpartum female rats. We also examined the possible behavioral mechanisms underlying the maternal effects of 5-HT1A. Sprague-Dawley mother rats were injected with a 5-HT1A agonist 8-OH-DPAT (0.1, 0.5 or 1.0 mg/kg, sc), a 5-HT1A antagonist WAY-101405 (0.1, 0.5 or 1.0 mg/kg, sc) or 0.9% saline solution on postpartum days 3, 5, and 7. Maternal behavior was tested 30 min before, 30 min, 120 min, and 240 min after the injection. Acute and repeated 8-OH-DPAT treatment significantly disrupted pup retrieval, pup licking, nursing, and nest building in a dose-dependent fashion, whereas WAY-101405 had no effect at the tested doses. The 5-HT1A receptor specificity of 8-OH-DPAT's action was confirmed as its maternal disruption effect was reversed by pretreatment of WAY-100635 (a highly selective 5-HT1A receptor antagonist). Subsequent pup preference test found that 8-OH-DPAT did not decrease the pup preference over a novel object, thus no inhibition on maternal motivation or maternal affect. The pup separation test and pup retrieval on an elevated plus maze test also failed to find any motivational and motor impairment effect with 8-OH-DPAT. However, 8-OH-DPAT at the maternal disruptive dose did disrupt the prepulse inhibition (a measure of attentional function) of acoustic startle response and enhanced the basal startle response. These findings suggest that stimulation of 5-HT1A receptors by 8-OH-DPAT impairs maternal care by partially interfering with the attentional processing or basal anxiety. More work is needed to further delineate the psychological and neuronal mechanisms underlying the maternal disruptive effect of 5-HT1A receptor activation.

Leonurine Exerts Antidepressant-Like Effects in the Chronic Mild Stress-Induced Depression Model in Mice by Inhibiting Neuroinflammation.

Background: Leonurine is an active alkaloid that is extracted from Traditional Chinese Medicine Herba leonuri. Emerging evidence indicates that leonurine produces neuroprotective effects in ischemic stroke, Parkinson's disease, and Alzheimer's disease. However, the effect of leonurine in neuropsychiatric disorders, especially in major depression, remains unknown. Methods: We used the chronic mild stress mouse model to explore the antidepressant effects of leonurine and the potential mechanisms. Behavioral tests including sucrose preference test, forced swimming test, and tail suspension test were taken to evaluate depression symptoms. Moreover, the contents of monoamine neurotransmitters in hippocampus and prefrontal cortex were measured by high-performance liquid chromatography. Neuronal morphology was detected by transmission electron microscopy. Results: Administration of leonurine (60 mg/kg) for 4 weeks significantly alleviated depression-like behaviors of chronic mild stress mice, including increased sucrose preference and reduced immobility time in forced swimming test and tail suspension test. We further found that leonurine (60 mg/kg) effectively restored the levels of 5-hydroxytryptamine, noradrenaline, and dopamine in the hippocampus and prefrontal cortex of chronic mild stress mice, accompanied by amelioration of hippocampal neuronal damage. Furthermore, leonurine (60 mg/kg) significantly inhibited the production of proinflammatory cytokine interleukin-1ß, interleukin-6 and TNF-α, and suppressed the nuclear factor kappa B signaling pathway. Conclusions: These findings demonstrate that leonurine exerts antidepressant-like effects, which may be mediated, at least in part, by improving monoamine neurotransmitters and inhibiting neuroinflammation. Our study provides insight into the potential of leonurine in depression therapy.

Time-dependent sensitization of antipsychotic effect in adolescent male and female rats.

Many behavioral and biological effects of a psychoactive drug often undergo time-dependent change following even one single drug exposure. The present study examined whether one or two exposures of haloperidol, olanzapine or clozapine would also induce a time-dependent change in their behavioral effects in adolescent rats, and whether such a change vary between sexes. Adolescent Sprague-Dawley rats (<40days old) were first treated with one single injection of haloperidol (0.05 and 0.1mg/kg, sc), clozapine (10.0 and 20.0mg/kg, sc), 2 injections of olanzapine (1.0 and 2.0mg/kg, sc) or vehicle, and tested in a conditioned avoidance response (CAR) model or a PCP (3.20mg/kg, sc)-induced hyperlocomotion model to assess the drug's antipsychotic-like behavioral effects. One or three weeks later, rats were challenged with the drug and their avoidance responses and the PCP-induced hyperlocomotion were re-assessed. One-trial haloperidol and 2-trial olanzapine induced a sensitization, while 1-trial clozapine induced a tolerance effect. The 1-trial haloperidol sensitization was significantly higher at the 3-week time point than at 1-week point, especially in the females. Clozapine tolerance in the conditioned avoidance response model also exhibited the time-dependent increase in both sex groups. Olanzapine sensitization in the PCP model showed a time-dependent change in a sex-dependent fashion. Overall, the time-dependent antipsychotic sensitization and tolerance can be demonstrated in adolescent animals. Many pharmacological (e.g. specific drugs, drug doses), individual (e.g. male versus female) and environmental (e.g. specific behavioral models) factors play a role in the modulation of the strength of antipsychotic sensitization and tolerance.

Quantitative EEG abnormalities in major depressive disorder with basal ganglia stroke with lesions in different hemispheres.

BACKGROUND: This study aimed to examine the aberrant EEG oscillation in major depressive subjects with basal ganglia stroke with lesions in different hemispheres. METHODS: Resting EEG of 16 electrodes in 58 stroke subjects, 26 of whom had poststroke depression (13 with left-hemisphere lesion and 13 with right) and 32 of whom did not (18 with left lesion and 14 with right), was recorded to obtain spectral power analysis for several frequency bands. Multiple analysis of variance and receiver operating characteristic (ROC) curves were used to identify differences between poststroke depression (PSD) and poststroke non-depression (PSND), treating the different lesion hemispheres separately. Moreover, Pearson linear correlation analysis was conducted to test the severity of depressive symptoms and EEG indices. RESULTS: PSD with left-hemisphere lesion showed increased beta2 power in frontal and central areas, but PSD with right-hemisphere lesion showed increased theta and alpha power mainly in occipital and temporal regions. Additionally, for left-hemisphere lesions, beta2 power in central and right parietal regions provided high discrimination between PSD and PSND, and for right-hemisphere lesions, theta power was similarly discriminative in most regions, especially temporal regions. We also explored the association between symptoms of depression and the power of abnormal bands, but we found no such relationship. LIMITATIONS: The sample size was relatively small and included subjects with different lesions of the basal ganglia. CONCLUSIONS: The aberrant EEG oscillation in subjects with PSD differs between subjects with lesions of the left and right hemispheres, suggesting a complex association between depression and lesion location in stroke patients.

Simultaneous determination of ten preservatives in ten kinds of foods by micellar electrokinetic chromatography.

An improved micellar electrokinetic capillary chromatography method (MEKC) for the simultaneous determination of ten preservatives in ten different kinds of food samples was reported. An uncoated fused-silica capillary with 50 µm i.d. and 70 cm total length was used. Under the optimized conditions, the linear response was observed in the range of 1.2-200mg/L for the analytes. The limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) ranging from 0.4 to 0.5mg/L and 1.2 to 1.5mg/L, respectively were obtained. The method was used for the determination of sorbic and benzoic acids in two FAPAS® (Food Analysis Performance Assessment Scheme) proficiency test samples (jam and chocolate cake). The results showed that the current method with simple sample pretreatment and small reagent consumption could meet the needs for routine analysis of the ten preservatives in ten types of food products.