Effects of mercury contaminated rice from typical chemical plant area in China on nitric oxide changes and c-fos expression of rats brain

China is one of countries with the highest mercury production in the world. The Guizhou Province in Southwestern China is currently one of the world's most important mercury production areas. In order to study the neurotoxicity of rice from Qingzhen Chemical Plant area and probe into the signal transduction molecular mechanism of injury in rat brain stimulation by mercury contaminated rice. The rats were exposed to mercury contaminated rice for 20 d. Both of the measurements of NO and NOS were processed according to the protocol of the kit. The effect of Hg contaminated rice on the expression of c-fos mRNA in rat brain and the expression of c-FOS protein in cortex, hippocampus were observed using reverse transcription polymerase chain reaction(RT-PCR) and immunocytochemical methods. The results showed the neural transmitter NO and NOS in brain were significantly change between exposure groups and control group; the mercury polluted rice induced significantly the expression of c-fos mRNA; the c-FOS positive cells in hippocampus and cortex of exposure groups were significant different from control group(p<0.01). It could be concluded that nitric oxide was involved in mercury contaminated rice induced immediate early gene c-fos expressions in the rat brain. Through food chain, local ecosystem and health of local people iave been deteriorated seriously by mercury. This serious situation will last a long period. In order to alleviate mercury pollution, more work needs to do.

Acupuncture therapy for experimental stomach ulcer and c-Fos expression in rats

AIM: To determine the role of acupuncture therapy in treating experimental gastric ulcer in rats.METHODS: Twenty-eight adult male Sprague-Dawley rats were randomly divided into four groups (pre-acupuncture group; acupuncture group; paradistance-acupuncture group;and control group), and pre-acupuncture, paradistanceacupuncture, and control groups received 5 μL acetic acid (200 mL/L HAc) injection after a same course of electroacupuncture (EA) treatment (4 Hz, 0.6 mA, 0.45 ms,45 min for 4 d). The rats in these three groups recovered within 4 d. The acupuncture group received EA therapy for 4 d, after HAc injection. The stomach was dissected to compare the pathological structures of ulcer. Also c-Fos activation in the nuclei of solitary tract (NTS) was observed under microscope after regular immunohistochemistry staining of brain stem sections.RESULTS: The number of ulcers was different among the four groups, especially between control group and paradistance-acupuncture group or pre-acupuncture group. In the latter group, the number of ulcers was much less. The gastric ulcer area was consistent with the histopathological results, indicating that pre-acupuncture had an obvious therapeutic effect on gastric ulcers.Acupuncture had a very modest effect and paradistanceacupuncture had no effect on gastric ulcers. No therapeutic effect was found in the control group. Fos-Li neurons in NTS induced by noxious gastric ulcer showed a significant difference between pre-acupuncture and control groups.CONCLUSION: Acupuncture before ulceration can obviously alleviate ulcer. The production of c-Fos proves that the vagus nerve mediates the induction of c-Fos in nuclei of solitary tract following experimental ulceration,suggesting that parasympathetic afferents promote the process of noxious visceral stimulation.

Effects of electroacupuncture on c-Fos expression in the spinal cord and brain of rats with chronic visceral hypersensitivity

BACKGROUND： Visceral hypersensitivity is the main cause of irritable bowel syndrome, c-Fos is a marker of visceral hypersensitivity in the central nervous system. Electroacupuncture can relieve chronic visceral hypersensitivity in rats, but the mechanism is still unknown. OBJECTIVE： To identify c-Fos expression in the spinal cord and cerebral cortex of rats with chronic visceral hypersensitivity, and to test the effects of electroacupuncture on pain sensitivity in rats with chronic visceral hypersensitivity. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at the Animal E：~perimental Center, Shanghai University of Traditional Chinese Medicine, from January to April, 2007. MATERIALS： A total of 24 neonatal, male, Sprague Dawley rats, aged five days old, were equally and randomly assigned into a normal group, a model group, and an electroacupuncture group. Rabbit anti-rat c-Fos antibody and Evision secondary antibody kits （Sigma, USA）, diaminobenzidine kit （Dako, Denmark）, and an LD202H electroacupuncture apparatus （Huawei, Beijing, China） were used in this study. METHODS： Neonatal rats from the model and electroacupuncture groups were used to establish rat models of chronic visceral hypersensitivity by the saccule stimulation method. After model establishment, 0.25 mm diameter electric needles were inserted into Tianshu （ST 25） and Shangjuxu （ST37） at a depth of approximately 0.5 cm, with an square wave （alternating current frequency at 100/20 Hz, amplitude ranged 0.2-0.6 ms, intensity at 1 mA） once for 20 minutes, once a day, for seven days. Rats in the normal and model groups were not treated. MAIN OUTCOME MEASURES： Following 7 days of treatment, c-Fos expression in the spinal cord and cerebral cortex was detected by immunohistochemistry. After the first electroacupuncture treatment, abdominal withdrawal reflex scores were investigated to evaluate the pain threshold for chronic visceral hypersensitivity in rats. RESULTS： Visceral hypersensitivity increased c-Fos staining （P 〈 0.05）, and electroacupuncture significantly decreased the number of these cells to near normal levels （P 〉 0.05）. Abdominal withdrawal reflex scores were significantly lower in the electroacupuncture and normal groups than in the model group （P 〈 0.05） and were similar between the electroacupuncture and normal groups （P 〉 0.05）. CONCLUSION： Electroacupuncture decreases c-Fos expression in the spinal cord and cerebral cortex and increases pain threshold in a chronic visceral hypersensitivity model in rats.

c-Fos expression within the L_5 spinal cord dorsal horn after spinal nerve ligation in rats Is intraplantar administration of glutamate different from intrathecal administration?

BACKGROUND： Injection of glutamate （Glu） in normal animals can cause neuronal c-Fos expression; however, whether Glu can induce spinal neuronal c-Fos expression in pain models is unclear. OBJECTIVE： To examine the effects of intraplantar and intrathecal injection of Glu on c-Fos expression in the L5 spinal cord dorsal horn Ⅰ/Ⅱ and Ⅲ/Ⅳ layers after spinal nerve ligation, and to study the effects of the N-methyI-D-aspartic acid （NMDA） receptor antagonist, D-2-amino-5-phosphonopentanoate （D-AP5）, and a selective group I mGluR antagonist, 7-hydroyiminocyclo propan[a]chromen-lacarboxylic acid ethyl ester （cpccoEt）. DESIGN, TIME AND SETTING： A randomized, controlled animal study was performed at the Department of Pharmacology, Oral Anatomy, and Neurobiology, Osaka University Graduate School of Dentistry, from December 2005 to December 2006. MATERIALS： Glu （5 μmol）, D-AP5 （50 nmot） and cpccoEt （250 nmol） were provided by Wako Pure Chemical Industries, Osaka, Japan, and diluted in saline （50 μL）. The pH of all solutions was adjusted to 7.4. METHODS： Twelve rats were randomly divided into sham operation （n = 6） and spinal nerve ligation （SNL; n = 6） groups for behavioral assessments of neuropathic pain after ligation surgery of the left L5-6 nerve segment. Another 60 rats were randomly divided into sham operation, SNL, saline-intraplantar, saline-intrathecal, Glu-intraplantar, Glu-intrathecal, D-AP5-intrathecal, Glu-D-AP5-intrathecal, cpccoEt-intrathecal, and Glu-cpccoEt-intrathecal groups, with 6 rats in each group. All groups except sham operation group received a similar SNL. On day 14, rats received a 50-μL injection of saline, Glu, D-AP5, and/or cpccoEt into the left intraplantar or intrathecal L5-4 segments. MAIN OUTCOME MEASURES： The number of c-Fos positive neurons in both Ⅰ/Ⅱ and Ⅲ/Ⅳ spinal layers at L6 was observed using immunohistochemistry 2 hours after administration. RESULTS： （1） SNL increased the level of c-Fos expression in two sides of the spinal cord, particularly on Ⅲ/Ⅳ spinal layers of the ligated side. （2） Intraplantar or intrathecal administration of saline significantly increased the c-Fos labeled neurons in Ⅰ/Ⅱ spinal layers of the ligated side, compared with SNL alone （P 〈 0.01）. （3） Intraplantar Glu （5 μmol） increased the number of c-Fos positive neurons in Ⅰ/Ⅱ spinal layers compared with intraplantar saline （P〈 0.01）. （4） The number of c-Fos neurons in Ⅰ/Ⅱ spinal layers on both the ipsilateral and contralateral side after intraplantar Glu was lower than intrathecal Glu （P〈 0.01）, with a 3-fold higher induction by intrathecal Glu. （5） Co-administration of D-AP5 or cpccoEt reduced the effects of intrathecal Glu （P 〈 0.01）. CONCLUSION： Intrathecal Glu increases c-Fos expression more than intraplantar Glu. Antagonists of NMDA and group I mGluRs block this effect.

Tegaserod inhibits noxious rectal distention induced responses and limbic system c-Fos expression in rats with visceral hypersensitivity

AIM: To examine the effects of tegaserod, a serotonin(5-HT) 4 receptor partial agonist, on abdominal withdrawalreflex (AWR) to rectal distention (RD) and c-Fos expressionin limbic system.METHODS: Neonatal Sprague-Dawley rats randomlyreceived colonic irritation by acetic acid from postnatal day8 to d 21 as a visceral hypersensitive model (group H) or byintrarectal saline as a control group (group C). When theybecame adults, rectal distention (RD) was performed by aballoon (6F; Fogarty arterial embolectomy catheter; length,20 mm; diameter, 2 mm) which was rapidly inflated withincreasing volumes of saline (0.4, 0.8 and 1.2 mL) for 20 sat five-minute intervals. Five subgroups of group H (H-saline,H-vehicle, H-Teg0.1, H-Teg0.3 and H-Tegl.0) were injectedrandomly with saline, vehicle (1-methyl-2-thpyrrolidone) ortegaserod at doses of 0.1, 0.3 and 1.0 mg/kg ip, respectively.Two subgroups of group C (C-Saline and C-Tegl.0) wereinjected with saline or tegaserod (1.0 mg/kg) ip. RD wasperformed 10 rain after injection, AWR was recorded andc-Fos expression in limbic system was analyzed quantitativelyby immunohistochemistry.RESULTS: Compared to saline, tegaserod significantlyinhibited AWR in group H (0.4 mL: from 2.0 to 0.5; 0.8 mL:from 3.5 to 1.5; 1.2 mL: from 4.0 to 3.0, P<0.01), but hadno significant effect on group C. Tegaserod dose-dependentlyattenuated the number of c-Fos positive neurons in limbicstructures, anterior cingulate cortex (ACC) showed thegreatest attenuation. In group H, tegaserod (1.0 mg/kg)resulted in a significant overall decrease to 57% of H-saline(283+41 vs 162+16, P<0.01), in ACC to 42% of H-saline(72+10 vs31+8, P<0.01). In group C, tegaserod (1.0 mg/kg)resulted in an overall decrease to 77% of C-saline (214+13vs 164+22, P<0.01), in ACC to 65% of C-saline (48+8 vs31+7, P<0.01).CONCLUSION: Tegaserod inhibits the response to rectaldistention in rats with visceral hypersensitivity and dose-dependently attenuates c-Fos expression in limbic system,especially in anterior cingulate cortex.

Protection of 3'-methoxy-puerarin against focal brain ischemia and its association with c-fos expression

The present study established a rat model of focal brain ischemia by occlusion of the middle cerebral artery covered with FeCl3, and investigated the protective effect of 3＇-methoxy-puerarin. Hippocampal and cortical c-fos gene expression was determined using in situ hybridization. Results showed that 3＇-methoxy-puerarin reduced neurological deficit scores, cerebral infarcted zone and water content of brain tissues, dramatically increased the activity of catalase and glutathione peroxidase in the ischemia zone of the hippocampus, increased the activity of catalase in the cortex, decreased lipid peroxide and lactic acid contents in the hippocampus and cerebral cortex, and down-regulated c-fos gene expression in brain ischemic rats. Results demonstrated that 3＇-methoxy-puerarin exhibited cerebroprotective effects against focal brain ischemia, which involved c-fos gene expression.

Effect of water restrictions on the physiological parameters,psychological behavior and brain c-Fos expression in rat

Objective In order to characterize the feature of stress response induced by stressor with both physical and psychological natures, the effect of water restriction performed in different experimental modes on the physiological parameters, psychological behavioral manifestations and brain c-Fos expressions were observed and compared. Methods 58 male Wistar rats were used and randomly divided into three experimental groups （n=18 for each） and a control group （n=4）. In control group, the rats were allowed to access drinking water freely at all experimental period. In the experimental groups the water supply to the rats was restricted. In timed water supply （TW） group, the water was supplied twice a day, 10 min for each in fixed hours every day. In empty bottle-served （EB） and water-restricted （WR） groups, the water was served only once a day for 10 min, either in the early morning or evening, and in the other time point scheduled for water supply only an empty bottle without water was provided in the EB group and nothing was given in the WR group. The quantities of drank water and eaten food, weight-gaining, and behavior score were observed every day. The serum level of corticosterone was assayed and the rats were sacrificed with fixative perfusion of 3 d, 7 d or 14 d respectively, following water restriction （n=6 for each time point in each group）. The brain c-Fos expressions were examined with immunohistochemistry. Results The slowing down of weight-gaining, rise of serum corticosterone level, occurrence of psychological behavioral manifestations of unpeaceful restlessness such as exploring and attacking, enhance of c-Fos expression in the subfomical organ （SFO）, median preoptic nucleus （MnPO）, area postrema （AP）, hypothalamic paraventricular nucleus （PVN）, supraoptic nucleus （SON）, medial （MeA） and central （CeA） amygdaloid nucleus and ventrolateral septum （LSV） were noticed in both EB and WR groups, except the nucleus of solitary tract （NTS） in which the Fos expression was decreased. The changes of Fos expression in most of nuclei in EB group began at day 3, at least persisted till day 7, and backed down at day 14. While in WR group, similar changes started at day 7 and reached its peak at day 14. In TW group, only the concentration of corticosterone at day 7 was slightly increased and the rest indexes observed were unchanged. Conclusion The results indicate that water restriction induces physical as well as psychological stress responses. And the water restrictions experimentally executed in different modes result in different manifestations of behavioral response and brain immediately early gene expression in discrete brain nuclei/regions.

四神丸对腹泻型肠易激综合征大鼠结肠MCT、c-fos表达的影响

目的观察四神丸对腹泻型肠易激综合征(IBS-D)模型大鼠结肠MCT、c-fos表达的影响。方法将40只大鼠随机分为正常组、模型组、匹维溴铵组(15.23 mg/kg)和四神丸组(7.32 mg/kg),每组10只,采用番泻叶灌胃联合避水应激法造模,灌胃给药14 d后,观察一般状态,测定体质量、粪便含水量和AWR评分,ELISA法检测血清MCT、c-fos水平,免疫组织化学法、Western blot法分别检测结肠组织MCT、c-fos蛋白定位及表达,RT-qPCR法检测结肠组织MCT、c-fos mRNA表达。结果与模型组比较,四神丸组和匹维溴铵组大鼠一般状态明显好转,体质量升高(P<0.01),粪便含水量、AWR评分以及血清中MCT、c-fos水平均降低(P<0.05,P<0.01),结肠组织MCT、c-fos蛋白及mRNA表达均降低(P<0.05,P<0.01);四神丸组结肠组织MCT蛋白表达及c-fos蛋白表达均低于匹维溴铵组(P<0.05)。结论四神丸对IBS-D大鼠内脏敏感的保护机制可能与调节结肠肥大细胞活化指标MCT、c-fos的表达有关。

艾灸对创伤后应激障碍小鼠行为学及下丘脑外侧区c-fos的影响及c-fos与行为学的相关性分析

目的观察艾灸“内关”“阳陵泉”对创伤后应激障碍(post traumatic stress disorder,PTSD)小鼠行为及小鼠下丘脑外侧区c-fos表达的影响。方法将C57小鼠随机分为正常组、模型组、艾灸组,每组6只。模型组、艾灸组采用改良的单次长时间应激和电刺激(single prolonged stress and electrical stimulation,SPS&S)方法复制PTSD模型。艾灸组予艾灸“内关”“阳陵泉”连续干预7 d。利用旷场实验、高架十字迷宫实验和条件性恐惧测试实验检测小鼠行为,采用免疫荧光检测小鼠下丘脑外侧区c-fos的表达水平。结果与正常组比较,模型组小鼠体质量,旷场实验中央场时间、中央场距离显著减少(P<0.05),高架十字迷宫实验开臂进入次数和时间均显著减少(P<0.05);条件性恐惧测试实验中,背景恐惧和声音恐惧的冻结时间均显著增加(P<0.05);小鼠下丘脑外侧区c-fos表达水平显著升高(P<0.05)。与模型组比较,艾灸组小鼠体质量,旷场实验中央场时间、中央场距离显著增加(P<0.05),高架十字迷宫实验开臂进入次数和时间均显著增加(P<0.05);条件性恐惧测试实验中,背景恐惧和声音恐惧的冻结时间均显著减少(P<0.05);小鼠下丘脑外侧区c-fos表达水平显著降低(P<0.05)。结论艾灸“内关”“阳陵泉”穴能够显著改善PTSD小鼠焦虑抑郁样行为,其机制可能与调节下丘脑外侧区的c-fos表达有关。

Astrocytic endothelin-1 overexpression impairs learning and memory ability in ischemic stroke via altered hippocampal neurogenesis and lipid metabolism

Vascular etiology is the second most prevalent cause of cognitive impairment globally.Endothelin-1,which is produced and secreted by endothelial cells and astrocytes,is implicated in the pathogenesis of stroke.However,the way in which changes in astrocytic endothelin-1 lead to poststroke cognitive deficits following transient middle cerebral artery occlusion is not well understood.Here,using mice in which astrocytic endothelin-1 was overexpressed,we found that the selective overexpression of endothelin-1 by astrocytic cells led to ischemic stroke-related dementia(1 hour of ischemia;7 days,28 days,or 3 months of reperfusion).We also revealed that astrocytic endothelin-1 overexpression contributed to the role of neural stem cell proliferation but impaired neurogenesis in the dentate gyrus of the hippocampus after middle cerebral artery occlusion.Comprehensive proteome profiles and western blot analysis confirmed that levels of glial fibrillary acidic protein and peroxiredoxin 6,which were differentially expressed in the brain,were significantly increased in mice with astrocytic endothelin-1 overexpression in comparison with wild-type mice 28 days after ischemic stroke.Moreover,the levels of the enriched differentially expressed proteins were closely related to lipid metabolism,as indicated by Kyoto Encyclopedia of Genes and Genomes pathway analysis.Liquid chromatography-mass spectrometry nontargeted metabolite profiling of brain tissues showed that astrocytic endothelin-1 overexpression altered lipid metabolism products such as glycerol phosphatidylcholine,sphingomyelin,and phosphatidic acid.Overall,this study demonstrates that astrocytic endothelin-1 overexpression can impair hippocampal neurogenesis and that it is correlated with lipid metabolism in poststroke cognitive dysfunction.

电针心经穴位对急性心肌缺血模型大鼠内侧隔核白细胞介素-2、JunB蛋白及c-fos表达水平的影响

目的观察电针心经对急性心肌缺血(acute myocardial ischemia,AMI)大鼠内侧隔核白细胞介素(interleukin,IL)-2水平、JunB蛋白及c-fos表达水平的影响,探究内侧隔核在针刺抗AMI中的作用及机制。方法将大鼠分为伪手术组、模型组和电针组,每组6只;结扎冠状动脉左前降支复制AMI大鼠模型;电针组大鼠选取手少阴心经“神门—通里”段进行干预,每次30 min,每日1次,连续电针3 d,刺激电流为1 mA,频率为2 Hz;伪手术组、模型组大鼠不进行电针干预。采用ELISA法检测大鼠大脑内侧隔核IL-2水平,Western blot法检测大鼠大脑内侧隔核区JunB蛋白表达水平,免疫荧光法检测大鼠大脑内侧隔核区c-fos免疫反应阳性神经元表达水平。结果与伪手术组比较,模型组大鼠大脑内侧隔核区IL-2、JunB蛋白水平显著升高(P<0.05),c-fos免疫反应阳性神经元数和平均吸光度(optical density,OD)值显著增加(P<0.05);与模型组比较,电针组大鼠大脑内侧隔核区IL-2、JunB蛋白水平显著降低(P<0.05),c-fos免疫反应阳性神经元数和平均OD值显著减少(P<0.05)。结论内侧隔核参与电针心经抗AMI的作用,其机制可能与电针降低大脑内侧隔核区IL-2、JunB蛋白及c-fos表达水平有关。

骨疏康干预破骨细胞:激活核因子E2相关因子2调控c-Fos/NFATc1通路

背景:已有研究表明,骨疏康通过调节核苷酸、氨基酸代谢和免疫机制影响骨骼代谢,目前骨疏康治疗骨质疏松症的机制研究主要聚焦于调控成骨细胞,对破骨细胞的关注较少。目的:以RAW 264.7细胞为实验对象,从破骨细胞角度探讨骨疏康治疗骨质疏松症的机制。方法:取8周龄雌性SD大鼠24只,采用随机数字表法分为4组(n=6),3个实验组分别灌胃给予1,2,4 g/kg的骨疏康药液(2次/d),对照组灌胃给予等量蒸馏水(2次/d),连续灌胃7 d后抽取大鼠主动脉血,离心收集血清,同组血清合并,获得低、中、高浓度的骨疏康含药血清及正常血清,进行后续实验。①将RAW 264.7细胞分6组培养:对照组加入正常血清,低、中、高浓度组分别加入低、中、高浓度的骨疏康含药血清,Nrf2抑制剂组加入核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)抑制剂ML385,Nrf2激活剂组加入Nrf2激活剂t-BHQ,采用CCK8法检测细胞相对活性。②将第3代RAW 264.7细胞分5组培养:空白对照组加入正常血清,破骨组加入核因子κB受体活化因子配体(receptor activator of nuclear factorκB ligand,RANKL),低、中、高浓度组在加入RANKL的基础上分别加入低、中、高浓度的骨疏康含药血清,培养5 d后进行抗酒石酸酸性磷酸染色。③将RAW 264.7细胞分5组培养:空白对照组加入正常血清,破骨组加入正常血清与RANKL,高浓度+破骨组加入RANKL+高浓度骨疏康含药血清,破骨+Nrf2激动剂组加入RANKL+t-BHQ,高浓度+破骨+Nrf2抑制剂组加入RANKL+高浓度骨疏康含药血清+ML385,培养5 d后进行Western Blot与活性氧含量检测。结果与结论:①CCK8检测结果显示,骨疏康含药血清及Nrf2抑制剂、激动剂对RAW 264.7细胞活力无明显影响;②抗酒石酸酸性磷酸染色结果显示,骨疏康含药血清呈浓度依赖性抑制破骨细胞的分化;③Western Blot与活性氧含量检测结果显示,与空白对照组比较,破骨组Nrf2蛋白表达降低(P<0.05),c-Fos、NFATc1蛋白表达与活性氧含量升高(P<0.05);与破骨组比较,高浓度+破骨组、破骨+Nrf2激动剂组、高浓度+破骨+Nrf2抑制剂组Nrf2蛋白表达升高、活性氧含量降低(P<0.05),高浓度+破骨组、破骨+Nrf2激动剂组c-Fos、NFATc1蛋白表达降低(P<0.05);与高浓度+破骨组比较,高浓度+破骨+Nrf2抑制剂组Nrf2蛋白表达降低(P<0.05),活性氧含量升高(P<0.05);④结果表明,骨疏康通过激活Nrf2减少活性氧生成,进而抑制下游c-Fos/NFATc1通路表达和破骨细胞分化。

Decoding Retinoblastoma: Differential Gene Expression

Background: Retinoblastoma, the most common intraocular pediatric cancer, presents complexities in its genetic landscape that necessitate a deeper understanding for improved therapeutic interventions. This study leverages computational tools to dissect the differential gene expression profiles in retinoblastoma. Methods: Employing an in silico approach, we analyzed gene expression data from public repositories by applying rigorous statistical models, including limma and de seq 2, for identifying differentially expressed genes DEGs. Our findings were validated through cross-referencing with independent datasets and existing literature. We further employed functional annotation and pathway analysis to elucidate the biological significance of these DEGs. Results: Our computational analysis confirmed the dysregulation of key retinoblastoma-associated genes. In comparison to normal retinal tissue, RB1 exhibited a 2.5-fold increase in expression (adjusted p Conclusions: Our analysis reinforces the critical genetic alterations known in retinoblastoma and unveils new avenues for research into the disease’s molecular basis. The discovery of chemoresistance markers and immune-related genes opens potential pathways for personalized treatment strategies. The study’s outcomes emphasize the power of in silico analyses in unraveling complex cancer genomics.

Effects of pyraclostrobin on growth,oxidative stress,and gene expression in relation to stress and ATP-binding cassette transporters in Tetrahymena thermophila

Pyraclostrobin(PYR),a widely used fungicide,has negative effects on fish and algae,but its toxicity in protozoa remains unclear.In this study,the effects of PYR on the growth,oxidative stress,and gene expression related to stress and ATP-binding cassette(ABC)transporters in Tetrahymena thermophila were investigated.The result showed that the 96-h IC_(50)of PYR against T.thermophila was 17.2 mg/L.Moreover,PYR inhibited the growth of T.thermophila in concentration-or time-dependent manner.A morphological study revealed that the shape and size of T.thermophila changed,and damage of cell membrane surface was observed by scanning electron microscopy after 96 h of PYR exposure.The activities of superoxide dismutase(SOD)and catalase(CAT)increased throughout the experiment.In contrast,the glutathione(GSH)content was increased at 24 h and 48 h of exposure and decreased at 96 h.Moreover,a significant increase in malondialdehyde(MDA)level was observed in T.thermophila after96 h of exposure.Furthermore,PYR upregulated the HSP703,HSP705,GPx2,and ABAC15 gene expression in the 0.1–5-mg/L groups and downregulated the HSP704,HSP90,TGR,and ABCC52 mRNA levels at 96 h of exposure.These results suggest that PYR may exert adverse effects on T.thermophila by inducing oxidative stress and changing the gene expression related to ABC transporters and stress,which may enrich the understanding of the toxicity mechanism of PYR in aquatic organisms and provide reference data for aquatic ecological risk assessments.

Effects of antioxidant‑rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats

Background Milk synthesis in lactating animals demands high energy metabolism,which results in an increased production of reactive oxygen metabolites(ROM)causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress(OS)on the animals.To mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation,a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus plantarum.Methods Twenty-four Guanzhong dairy goats(38.1±1.20 kg)were randomly assigned to two dietary treatments:one containing silage inoculated with L.plantarum MTD/1(RSMTD-1),and the other containing silage inoculated with high antioxidant activity L.plantarum 24-7(ES24-7).Results ES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to RSMTD-1.The ES24-7 diet elevated the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione peroxi-dase(GSH-Px),and catalase(CAT)activities in milk,serum,and feces of lactating goats(with the exception of T-AOC in milk).Additionally,the diet containing ES24-7 inoculated silage enhanced casein yield,milk free fatty acid(FFA)content,and vitamin A level in the goats’milk.Furthermore,an increase of immunoglobulin(Ig)A,IgG,IgM,inter-leukin(IL)-4,and IL-10 concentrations were observed,coupled with a reduction in IL-1β,IL-2,IL-6,interferon(IFN)-γ,and tumor necrosis factor(TNF)-αconcentrations in the serum of lactating goats fed ES24-7.Higher concentrations of total volatile fatty acid(VFA),acetate,and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated silage.Moreover,the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2(NFE2L2),beta-carotene oxygenase 1(BCO1),SOD1,SOD2,SOD3,GPX2,CAT,glu-tathione-disulfide reductase(GSR),and heme oxygenase 1(HMOX1)genes in the mammary gland,while decreased the levels of NADPH oxidase 4(NOX4),TNF,and interferon gamma(IFNG).Conclusions These findings indicated that feeding L.plantarum 24-7 inoculated alfalfa silage not only improved rumen fermentation and milk quality in lactating dairy goats but also boosted their immunity and antioxidant status by modulating the expression of several genes related to antioxidant and inflammation in the mammary gland.

Synergy of gut microbiota and host genome in driving heterosis expression of chickens

Heterosis has been widely utilized in agricultural production.Despite over a century of extensive research,the underlying mechanisms of heterosis remain elusive.Most hypotheses and research have focused on the genetic basis of heterosis.However,the potential role of gut microbiota in heterosis has been largely ignored.Here,we carefully design a crossbreeding experiment with two distinct broiler breeds and conduct 16S rRNA amplicon and transcriptome sequencing to investigate the synergistic role of gut microbiota and host genes in driving heterosis.We find that the breast muscle weight of hybrids exhibits a high heterosis,6.28%higher than the mid-parent value.A notable difference is observed in the composition and potential function of cecal microbiota between hybrids and their parents.Over 90%of differentially colonized microbiota and differentially expressed genes exhibit nonadditive patterns.Integrative analyses uncover associations between nonadditive genes and nonadditive microbiota,including a connection between the expression of cellular signaling pathways and metabolism-related genes and the abundance of Odor-ibacter,Oscillibacter,and Alistipes in hybrids.Moreover,higher abundances of these microbiota are related to better meat yield.In summary,these findings highlight the importance of gut microbiota in heterosis,serving as crucial factors that modulateheterosis expression in chickens.

Molecular mechanism of hypoxia and alpha-ketoglutaric acid on collagen expression in scleral fibroblasts

AIM:To investigate the molecular mechanisms underlying the influence of hypoxia and alpha-ketoglutaric acid(α-KG)on scleral collagen expression.METHODS:Meta-analysis and clinical statistics were used to prove the changes in choroidal thickness(ChT)during myopia.The establishment of a hypoxic myopia model(HYP)for rabbit scleral fibroblasts through hypoxic culture and the effects of hypoxia andα-KG on collagen expression were demonstrated by Sirius red staining.Transcriptome analysis was used to verify the genes and pathways that hypoxia andα-KG affect collagen expression.Finally,real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used for reverse verification.RESULTS:Meta-analysis results aligned with clinical statistics,revealing a thinning of ChT,leading to scleral hypoxia.Sirius red staining indicated lower collagen expression in the HYP group and higher collagen expression in the HYP+α-KG group,showed that hypoxia reduced collagen expression in scleral fibroblasts,whileα-KG can elevated collagen expression under HYP conditions.Transcriptome analysis unveiled the related genes and signaling pathways of hypoxia andα-KG affect scleral collagen expression and the results were verified by RT-qPCR.CONCLUSION:The potential molecular mechanisms through which hypoxia andα-KG influencing myopia is unraveled and three novel genes TLCD4,TBC1D4,and EPHX3 are identified.These findings provide a new perspective on the prevention and treatment of myopia via regulating collagen expression.

A Facial Expression Recognition Method Integrating Uncertainty Estimation and Active Learning

The effectiveness of facial expression recognition(FER)algorithms hinges on the model’s quality and the availability of a substantial amount of labeled expression data.However,labeling large datasets demands significant human,time,and financial resources.Although active learning methods have mitigated the dependency on extensive labeled data,a cold-start problem persists in small to medium-sized expression recognition datasets.This issue arises because the initial labeled data often fails to represent the full spectrum of facial expression characteristics.This paper introduces an active learning approach that integrates uncertainty estimation,aiming to improve the precision of facial expression recognition regardless of dataset scale variations.The method is divided into two primary phases.First,the model undergoes self-supervised pre-training using contrastive learning and uncertainty estimation to bolster its feature extraction capabilities.Second,the model is fine-tuned using the prior knowledge obtained from the pre-training phase to significantly improve recognition accuracy.In the pretraining phase,the model employs contrastive learning to extract fundamental feature representations from the complete unlabeled dataset.These features are then weighted through a self-attention mechanism with rank regularization.Subsequently,data from the low-weighted set is relabeled to further refine the model’s feature extraction ability.The pre-trained model is then utilized in active learning to select and label information-rich samples more efficiently.Experimental results demonstrate that the proposed method significantly outperforms existing approaches,achieving an improvement in recognition accuracy of 5.09%and 3.82%over the best existing active learning methods,Margin,and Least Confidence methods,respectively,and a 1.61%improvement compared to the conventional segmented active learning method.

Global-local combined features to detect pain intensity from facial expression images with attention mechanism

The estimation of pain intensity is critical for medical diagnosis and treatment of patients.With the development of image monitoring technology and artificial intelligence,automatic pain assessment based on facial expression and behavioral analysis shows a potential value in clinical applications.This paper reports a framework of convolutional neural network with global and local attention mechanism(GLA-CNN)for the effective detection of pain intensity at four-level thresholds using facial expression images.GLA-CNN includes two modules,namely global attention network(GANet)and local attention network(LANet).LANet is responsible for extracting representative local patch features of faces,while GANet extracts whole facial features to compensate for the ignored correlative features between patches.In the end,the global correlational and local subtle features are fused for the final estimation of pain intensity.Experiments under the UNBC-McMaster Shoulder Pain database demonstrate that GLA-CNN outperforms other state-of-the-art methods.Additionally,a visualization analysis is conducted to present the feature map of GLA-CNN,intuitively showing that it can extract not only local pain features but also global correlative facial ones.Our study demonstrates that pain assessment based on facial expression is a non-invasive and feasible method,and can be employed as an auxiliary pain assessment tool in clinical practice.

Genome-wide identification and expression profiling of photosystem II(PsbX)gene family in upland cotton(Gossypium hirsutum L.)

Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.