ACTA HISTOCHEMICA
·
2021
·
IF 2.479
Xian-Zhou Feng,Wei-Yong Wang,Ting Gong,Li-Jie Meng
The optimized fixative for testis is still controversial. This study investigated the effects of Modified Davidson’s Fluid (mDF), 4% Paraformaldehyde (4% PFA), and Bouin’s Fluid (BF) fixatives on chicken testes in normal/cadmium (Cd) feeding groups using hematoxylin and eosin (HE), immunohistochemistry (IHC), and Terminal Transferase dUTP Nick End Labeling (TUNEL) staining. Compared to the mDF, we established that the testes fixed with 4% PFA and BF in the normal group had severe shrinkage in tubular and interstitial compartments. Moreover, compared with 4% PFA, the number of GATA4-positive Sertoli cells/mm 2 reduced by 67.61% in mDF and 80.57% in BF for one seminiferous tubule . The TUNEL assay illustrated that more positive cells/mm 2 in mDF group (28.47 ± 11.38) than in 4% PFA (10.49 ± 7.89). In Cd-treated testes, mDF showed more morphological details than 4% PFA and BF. In contrast, the number of GATA4-positive Sertoli cells/mm 2 of 4% PFA was higher than that of mDF by 65.78% and BF by 64.80% in a seminiferous tubule. The number of TUNEL positive cells/mm 2 in mDF (272.60 ± 34.41) were higher than in 4% PFA (175.91 ± 19.87). These results suggest that mDF fixative is suitable for normal and Cd-treated testis fixation for HE and TUNEL staining in chicken, whereas 4% PFA fixative is better for IHC examination.
研究领域:
组织学
·
毒理学
·
生殖生物学
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产品关联
DOI
PubMed
高影响因子
Molecular Medicine Reports
·
2019
·Vol. 19
·期 4
·2758-2766
·
IF 1.851
Ya Hou,Xiaobo Wang,Xiaorui Chen,Jing Zhang,Xiaopeng Ai,Yusheng Liang,Yangyang Yu,Yi Zhang,Xianli Meng,Tingting Kuang,Yao Hu
This study was conducted to establish a stable hypobaric hypoxia brain injury model. SD rats were randomly separated into control and model groups, and placed outside or inside of a hypobaric chamber, respectively. Subsequent to 24 h anoxic exposure, plasma superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSG) and lactate dehydrogenase (LDH) were measured using commercial biochemical kits. Hematoxylin‑eosin (H&E), Nissl's and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were used to observe the morphology of neurons in the hippocampus. The protein expression levels of apoptotic protease activating factor‑1 (Apaf‑1), hypoxia inducible factor‑1α (HIF‑1α), caspase‑3, cleaved caspase‑3, Bcl‑2‑associated X protein (Bax) and cytochrome c (cyto‑c) were detected using western blot and immunohistochemistry analyses. Hypoxic substantially induced morphological lesions in the hippocampus concomitant with the physical behavioral performance deficit. Furthermore, hypoxia markedly exacerbated the levels of MDA, LDH and GSSG, and restrained GSH (P<0.01) and SOD (P<0.05) levels compared with the control group. In addition, hypoxia significantly induced the protein expression of Apaf‑1, HIF‑1α, caspase‑3, cleaved caspase‑3, Bax and Cyto‑c (P<0.01) compared with the control group. Finally, a lower number and volume of Nissl bodies were verified in the hypoxic group. TUNEL results demonstrated a greater number of apoptotic cells in the hypoxic group. The present study demonstrates a model of rat hypoxic brain injuries induced by a hypobaric chamber at 9,000 m for 24 h. Furthermore, the redox enzyme, HIF‑1α and mitochondrial apoptosis‑associated protein, along with H&E and Nissl's staining, may be applied to evaluate the degree of injury.
研究领域:
神经科学
·
药理学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
BRITISH POULTRY SCIENCE
·
2025
·
IF 1.7
Y. S. Miao,Q. X. Han,K. X. Wang,X. D. Fan,Z. Zhang,L. Chen,D. Z. Zheng,S. Yue,L. Lei,L. Y. Liu,J. C. Li,G. J. Liu
1. This study evaluated the mechanism of action of indirubin in alleviating the structural damage induced by Mycoplasma gallisepticum (MG) in the lungs and trachea.2. A total of 250 one-day-old white leghorn chickens, specific-pathogen-free were divided into six treatment groups, including (A) indirubin high concentration treatment group (50 mg/kg; IHC); (B) indirubin medium concentration treatment group (25 mg/kg; IMC); (C) indirubin low concentration treatment group (12.5 mg/kg; ILC); (D) tylosin control group (0.5 g/l); (E) control group (CON) and (F) challenge model group (MG).3. Results from antioxidant activity analysis demonstrated that indirubin treatment significantly decreased the amount of MG-mediated oxidative stress in the lungs of chickens. Histopathological examination revealed abnormal morphological signs and cell damage in MG birds. This included lung lymphocytic infiltration, overlapping nuclear debris and inflammatory cell infiltration. In addition, ultrastructural examination revealed signs of apoptosis in the lungs. However, indirubin treatment partially relieved these abnormal morphological changes.4. The TUNEL analysis showed extensive apoptosis in the lungs of the model group compared to the control and positive drug control group. Apoptosis-related protein expression levels were significantly upregulated in the model group, which confirmed the phenomena of apoptosis induced by MG. The indirubin treatment significantly reduced apoptosis in the lungs and trachea compared to the model group. Meanwhile, the effect of MG challenge was reduced in the lungs by indirubin in a dose-dependent manner.5. These results showed that the inhibition of oxidative stress and apoptosis by indirubin contributed to its therapeutic effects against MG infection.
研究领域:
兽医学
·
药理学
·
微生物学
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病理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2022
·
IF 8.786
Liu Senyue,Luo Lin,Zuo Fengyuan,Geng Yi,Ou Yangping,Chen Defang,Yang Shiyong,Luo Wei,Wang Yan,Wang Jun,Huang Xiaoli
Myxobolus, a major harmful type of myxospora, is one of the main parasitic pathogens of freshwater fish. Once myxoboliosis occurs, treatment can be extremely difficult. Therefore, clear understandings of the etiology of myxoboliosis and its pathological mechanism are keys for prevention and control. Here, histology, transmission electron microscopy, transcriptome study, tunel assay, and immunohistochemistry were carried out, revealing the morphology, pathological effects as well as host response mechanism of goldfish gill to Myxobolus ampullicapsulatus. Histological studies showed that the mature spores of Myxobolus ampullicapsulatus were composed of three parts, the spore shell, sporoplasm and bottle shaped polar capsule containing double S-shaped polar filaments. Transcriptome analysis revealed that Myxobolus ampullicapsulatus -infected (Myx) goldfish gills were characterized by apoptosis activation mediated by “p53 signaling pathway” with significantly up-regulated apoptosis-related differential genes dominated by p53-Bcl2/Bax signaling pathway. In addition, tunel assay revealed severe gill apoptosis in the Myx group. Transcriptome analysis also revealed that Myx group showed changes in immune response and significantly down-regulated immune-related differential genes. Beyond that, immunohistochemistry showed that there was no significant increase in the number of gill lymphocyte after parasite infection. These results suggest that the pathological mechanism of Myxobolus ampullicapsulatus infection on gills of goldfish may be related to apoptosis and immunosuppression. Subsequent qRT-PCR showed that apoptosis-related genes (Caspase3,Bad, Bax) and anti-inflammatory gene IL-10 were significantly increased, while immune-related pro-inflammatory genes (IL-1β, IL-8) were markedly down-regulated, further verifying the transcriptome results. Based on the above results, we concluded that p53-Bcl2/Bax related networks that dominant the expression of apoptosis genes were activated while immunity was suppressed in the gills of Myxobolus ampullicapsulatus infected goldfish. Our study is not only of benefit to enrich the taxonomy of Myxobolus but also clarifies its pathogenic mechanism, thus providing targets for prevention and control of myxoboliosis.
研究领域:
鱼类病理学
·
分子生物学
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寄生虫学
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免疫学
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转录组学
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产品关联
DOI
PubMed
高影响因子
NEUROENDOCRINOLOGY
·
2023
·Vol. 113
·期 6
·641-656
·
IF 4.1
Zheng Biaolin,Chen Xiubing,Chen Fengping,Liao Xiaomin,Wang Feng,Yang Zhe,Yi Nan,Shi Xiaoyan,Qin Shanyu,Jiang Haixing
Introduction: 5-Iodotubercidin, a type of purine derivative, has attracted increasing attention in tumor chemotherapy because of its potential as an antitumor agent in recent years. In this study, we confirmed the effects on apoptosis in insulinoma cell lines induced by 5-iodotubercidin and tried to illuminate the underlying mechanisms. Methods: We used 5-iodotubercidin in the treatment of insulinoma cells and the cell proliferation was examined using CCK-8 assay, colony-forming assays, and insulinoma animal models. Cell apoptosis was examined using TUNEL assays and Western blotting. Cellular DNA damage was shown by comet assay and immunofluorescence. The expression of apoptosis-regulating proteins and DNA damage biomarker was investigated by Western blotting. Subcutaneous inoculation of the insulinoma cells into nude mice was to measure blood glucose, insulin levels, and tumor growth. ATM siRNA and p53 siRNA were used as loss-of-function targets to evaluate 5-iodotubercidin treatment. Results: 5-Iodotubercidin inhibited the proliferation of insulinoma cells and induced DNA damage and cell apoptosis. Moreover, 5-iodotubercidin induced ATM and p53 activated. In vivo, 5-iodotubercidin inhibited the growth of Ins-1 and Min-6 cells xenografts in nude mice. Conclusion: 5-Iodotubercidin induces DNA damage leading to insulinoma cells apoptosis by activating ATM/p53 pathway. Therefore, this is a potential strategy for treating insulinoma
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
·
药理学
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DOI
PubMed
高影响因子
Experimental and Therapeutic Medicine
·
2017
·Vol. 15
·期 1
·897-903
·
IF 1.261
Jing Chen,Chenli Yang,Xiang Xu,Yonglin Yang,Bo Xu
The present study analyzed the change of Toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) expression in focal cerebral ischemia-reperfusion injury model. A sample of 36 Sprague-Dawley rats were randomly selected and divided into sham operation group (group S), control group (group C) and Chrysanthemum ester group (NF-κB inhibitor, group CE), each group consisted of 12 rats. The rat model of focal cerebral ischemia-reperfusion was established. The physiological indexes and neurological severity score of rats was recorded by a double-blind method. The cerebral infarction area was evaluated by triphenyltetrazole oxide (TTC) staining on brain slices. Apoptosis was evaluated by TUNEL staining. Semi-quantitative PCR and western blot analysis was used to measure the expression of TLR4 and NF-κB. The neurological severity score of rats in group C and CE were found to be significantly lower than group S (P<0.01). The TTC staining results showed that group C and CE had different levels of cerebral infarction but the area of infarction in group CE was significantly lower than group C (P<0.01). In addition, the number of TUNEL positive cells in group CE was significantly lower than group C (P<0.01). Semi-quantitative PCR and westernblot analysis results showed that the expression of NF-κB and TLR4 of group S was significantly lower than that of group C and group CE (P<0.01), the relative expression of NF-κB and TLR4 of group CE was significantly lower than that of group C (P<0.01). Moreover, the expression of NF-κB p65/p50 of group CE and group C was significantly higher than that of group S (P<0.01). This study concludes that the focal cerebral ischemia-reperfusion injury in rats can cause brain damage and cell apoptosis. This effect might be associated to the increased expression of NF-κB and TLR4, and the activation of TLR4/NF-κB signaling pathway.
研究领域:
神经科学
·
分子生物学
·
药理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF ALZHEIMERS DISEASE
·
2016
·Vol. 50
·期 4
·1149-1162
·
IF 3.92
Zhu Mingming,Huang Cong,Ma Xiao,Wu Rui,Zhu Weiwei,Li Xiaoting,Liang Zhaofeng,Deng Feifei,Zhu Jianyun,Xie Wei,Yang Xue,Jiang Ye,Wang Shijia,Wu Jieshu,Geng Shanshan,Xie Chunfeng,Zhong Caiyun
Neuronal cell death is an important feature of neurodegeneration. Aluminum is associated with neurodegenerative disorders, particularly Alzheimer’s disease. However, the underlying mechanisms by which aluminum induces neuronal apoptosis remain to be elucidated. miR-19 is a key miRNA implicated in regulating cell survival process, while the role of miR-19 in Alzheimer’s disease has not been investigated. In the present study, we showed that Aluminum maltolate (Al-malt), a lipophilic Al complex which is a common component of human diet with the ability to facilitate the entry of Al into the brain, induced apoptosis in human neuroblastoma SH-SY5Y cells, along with downregulation of miR-19a/miR-19b, upregulation of miR-19-targeted PTEN, and alterations of its downstream apoptosis related proteins including AKT, p53, Bax, and Bcl-2. miR-19 overexpression attenuated Al-malt-induced apoptosis as well as changes in the expression of apoptosis related proteins in SH-SY5Y cells. We further revealed that exposure of rats to Al-malt for 12 weeks at doses relevant to human exposure significantly elevated Al concentrations in serum and brain tissues. Al-malt dose-dependently induced apoptosis in rat brain, as evidenced by increased caspase activation and increased TUNEL staining. Consistent with in vitro results, Al-malt reduced miR-19 expression and altered the expression of apoptotic related proteins in rat brain. Taken together, our data suggest for the first time that miR-19 modulation is critically involved in Al-induced neural cell apoptosis. Findings from this study could provide new insight into the molecular mechanisms of Al-associated neurodegenerative pathogenesis.
研究领域:
神经科学
·
分子生物学
·
毒理学
·
神经退行性疾病
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产品关联
DOI
PubMed
高影响因子
PHYTOMEDICINE
·
2021
·
IF 5.34
Shusheng Fan,Xiaoyun Liu,Yu Wang,Xueyang Ren,Yue Liu,Ying Dong,Qiqi Fan,Jing Wei,Jiamu Ma,Axiang Yu,Ruolan Song,Hong Sui,Meng Shen,Fang Fang,Qing Xia,Gaimei She
Background Thymus quinquecostatus Celak. has been widely used as a spice and a folk medicine for relieving exterior syndrome and alleviating pain in China. Purpose To explore the protective effects and the underlying mechanism against cerebral ischemia-reperfusion injury (CIRI) of the T. quinquecostatus combining with its chemical composition. Study design and methods High-polar extract (HPE) was extracted from T. quinquecostatus and polyphenols in HPE were enriched to obtain polyphenol-rich fraction (PRF) using Macroporous resin. The free radicals and zebrafish embryos were used to compare the antioxidant activities of HPE and PRF in vitro and in vivo . Then, the transient middle cerebral artery occlusion (tMCAO) model was established in rats. Neurological deficit score, infarction rate, morphology and apoptosis of neurons were examined to investigate the protective effects of PRF on CIRI. The mRNA and protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1) and the activities of downstream antioxidant enzymes in ischemia tissues were determined to clarify the underlying mechanisms. Also, reactive oxygen species (ROS) level in zebrafish embryos were detected after incubation with PRF for a short time (2 h) to investigate whether PRF could directly eliminate free radicals. Finally, chemical composition of PRF were analyzed to investigate the material basis for antioxidant activity and anti-CIRI effect. Results Compared with HPE, PRF showed stronger antioxidant activities. PRF exhibited obvious protective effects including ameliorating neurological deficit, lowering infarction rate, and improving the cellular morphology in hippocampus CA1 and cortex after tMCAO. TUNEL staining suggested PRF dose-dependently improved the apoptosis of the neurons in ischemic cortex. RT-qPCR and Western Blot results suggested that PRF regulated oxidative stress (OS) via activating the Keap1/Nrf2/HO-1 signaling pathway . Also, PRF could directly scavenge excessive ROS in zebrafish embryos after a short-time PRF incubation. The anti-CIRI effect might be primarily attributed to the abundant polyphenols in PRF, including flavonoids , polymethoxylated flavonoids, flavonoid glycosides , and phenolic acids. Conclusion T. quinquecostatus contains abundant polyphenols and exhibited a good protective effect against CIRI via dual antioxidant mechanisms, providing a reference for further research and application for this plant.
研究领域:
药理学
·
神经科学
·
分子生物学
·
植物科学
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产品关联
DOI
PubMed
高影响因子
BIOLOGICAL TRACE ELEMENT RESEARCH
·
2025
·
IF 3.6
Duan Yunping,Liu Hui,Zhang Weichuan,Pei Zhuo,Huang Yong
In this study, 360 healthy juvenile largemouth bass, averaging body weights of 11.26 ± 0.49 g, were randomly assigned to four experimental groups. These groups were exposed to water environments containing different concentrations of cadmium (Cd): 0 mg/L for the control (Con) group, 0.9804 mg/L for the low-cadmium (LCd) group, 1.9608 mg/L for the medium-cadmium (MCd) group, and 9.804 mg/L for the high-cadmium (HCd) group, over a duration of 21 days, to study how different levels of Cd impact the liver. Ultrastructural changes manifest as nuclear deformation, blurred cell membrane boundaries, mitochondrial swelling, matrix dissolution, cristae rupture, and disappearance. TUNEL analysis indicates that the apoptotic rate of hepatocytes increases with increasing Cd concentration. Compared with the Con group, serum biochemical results showed that the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glucose (GLU), and urea (UREA) were significantly elevated in the HCd group. With increasing Cd concentration, the activities of hepatic catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH) were significantly altered, accompanied by a reduction in ATP content and an increase in the malondialdehyde (MDA) levels. Analysis of inflammatory gene expression showed upregulation of pro-inflammatory genes (IL-1β, TNF-α, and IL-6) and downregulation of the anti-inflammatory gene IL-10. In addition, significant changes in the mRNA levels of apoptosis-related genes were observed in these Cd-exposed groups. Collectively, these findings consistently indicate that Cd exposure can lead to liver injury in largemouth bass by inducing oxidative stress, inflammatory responses, and apoptosis. Graphical
研究领域:
环境毒理学
·
水生毒理学
·
生物化学
·
分子生物学
·
生态毒理学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 16.744
Yun Zeng,Yonghua Zhan,Xinyue Liu,Jingwen Ma,Huifang Liu,Hanrui Li,Tong Yi,Qingxia Zhu,Getao Du,Linfei Zhao,Dan Chen,Xueli Chen
Tumor metastasis is a major cause of poor cancer prognosis. Photothermal therapy (PTT) is a non-invasive, non-oxygen-dependent, and highly spatiotemporally precise treatment that may aid chemotherapy in suppressing tumor growth and metastasis. This article aims to develop and modify a CuS-based nanoplatform linking RGD-Acrk to actively target and internalize doxorubicin (DOX) into tumor cells, resulting in an alternating strategy of synergistic chemo/PTT. The nanoparticle’s shape, spectra, drug encapsulation efficiency, loading content, and controlled release are characterized, and a comparison of 808/980 nm lasers is used to irradiate the nanoplatform. The cell survival rate is used to determine the cytotoxicity of DOX and PTT, and the chemo/PTT is used to treat 4T1-Luc tumor-bearing mice. Simultaneously, bioluminescence and photoacoustic imaging are employed to assess therapy efficacy, as well as tumor size, weight, density, the bodyweight of mice, and survival curves. H&E, immunohistochemistry for HIF-1α, and TUNEL fluorescence staining confirm these findings, suggesting that the nanoplatform has a good active-targeted capacity and perform high-efficiency PTT caused by 980 nm laser irradiation. The synergistic effect is at least 3.53 times higher than chemo/PTT without synergistic effect and tumor liver metastasis is successfully suppressed, showing that it has the potential to be used for cancer treatment and prognosis improvement.
研究领域:
肿瘤学
·
纳米技术
·
诊疗学
✓ 已验证
DOI
高影响因子
NEUROSCIENCE
·
2018
·
IF 3.382
Lingqi Gao,Junde Han,Jie Bai,Jing Dong,Sen Zhang,Mazhong Zhang,Jijian Zheng
Early synchronized spontaneous network activity is a hallmark of the brain growth spurt period, during which general anesthetics cause widespread neuronal apoptosis and subsequent cognitive dysfunction. However, the relationship of such activity to anesthetic-induced neuronal apoptosis remains to be determined. In this study, we utilized patch-clamp electrophysiological recording, immunohistochemistry, and TUNEL assays to investigate the potential roles of spontaneous network activity in ketamine-induced neuronal apoptosis during early development. All experiments were performed using acutely dissected whole-mount Sprague–Dawley rat retinas (0–14 postnatal days [P0–P14]). Ketamine reversibly blocked spontaneous network activity in the rat retina from P0 to P9 and irreversibly blocked such activity from P10 to P12. The peak of physiological and ketamine-induced neuronal apoptosis mainly occurred from P7 to P9. Blockade of nicotinic acetylcholine receptors (nAChRs) also induced reversible inhibition of spontaneous network activity from P0 to P7 and extensive neuronal apoptosis in the P7 rat retina, while activation of nAChRs or increases in endogenous ACh levels attenuated ketamine-induced apoptotic responses. Furthermore, blockade of α7-nAChR and ß2-nAChR subtypes induced neuronal apoptosis in the developing retina, while activation of the α7-nAChR subtype attenuated ketamine-induced apoptotic responses. These results demonstrate that ketamine may inhibit early synchronized spontaneous network activity by blocking nAChRs, and that such inhibition may contribute to ketamine-induced neuronal apoptosis in the developing rat retina.
研究领域:
神经科学
·
药理学
·
发育生物学
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2024
·
IF 5.4
Mingyue Ruan,Xinran Wang,Mingxue Guo,Zucheng Hu,Wenjun Hu,Zishuo Guo,Wanling Chen,Shiyan Li,Kai Wu,Shouying Du,Ning Han
Combined chemo-phototherapy has shown considerable advantages and potential in cancer treatment. For this purpose, self-assembled nanoparticles by gambogic acid (GA) and IR780 (referred to as GA-IR780 NPs) were prepared. Herein, GA, an active compound derived from Garcinia hanburyi Hook.f, was selected as a chemo-agent. IR780 was used as a photothermal agent as well as a photosensitizer, which could kill tumor cells via photothermal effect and photodynamic effect. The obtained GA-IR780 NPs were uniform spheres with particle size of ca. 50 nm. The drug loading efficiency of GA and IR780 was 38.42 % and 56.64 %, respectively. The GA-IR780 NPs exhibited excellent photothermal properties as well as photodynamic effect when irradiated by near infrared (NIR) light (808 nm, 2.0 W/cm 2 ). Moreover, the GA-IR780 NPs showed enhanced cytotoxicity with NIR light activation. Results of animal experiments showed that GA-IR780 NPs had the most significant tumor inhibition when irradiated by laser, and the results of H&E , Ki-67 and TUNEL staining confirmed that the GA-IR780 NPs+Laser group caused the most severe tumor tissue damage. The above results indicated that GA-mediated chemotherapy combining with IR780-based phototherapy could significantly improve the anti-tumor efficacy.
研究领域:
药学
·
癌症研究
·
纳米技术
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）