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D-galactose (D-gal) is a reducing sugar widely distributed in food. In a pregnant animal model exposed to D-gal, D-gal was found to have toxic effects on both the mother and foetus through oxidative stress. However, little is known about the effect of D-gal exposure on the placenta and its underlying mechanism. In this study, we evaluated the effects of D-gal on HTR8/SVneo cells and the mechanisms in vitro. In the present study, the activity of HTR8/SVneo human trophoblasts decreased in a time- and concentration-dependent manner after exposure to D-gal. D-gal resulted in premature senescence of HTR8/SVneo cells, as confirmed by assessing β-galactosidase (SA-β-gal) activity and the expression of senescence-related factor p21. We also verified the damage of oxidative stress induced by D-gal by measuring the expression of reactive oxygen species (ROS), sirtuin 1 (SIRT1) and forkhead box O (FOXO) 3a. SRT1720, as a SIRT1 activator, mitigated D-gal-induced oxidative stress and senescence by upregulating SIRT1 and FOXO3a expression and reducing ROS production. Our data suggest that D-gal may induce HTR8/SVneo premature ageing through the SIRT1/FOXO3a/ROS signalling pathway mediated by oxidative stress and that SIRT1 protects cells from this damage.

Cellular senescence is a key factor in the development of intervertebral disc degeneration (IVDD). Age-associated decreases in NAD+ levels play a critical role in regulating cellular senescence. Previous studies have found that small extracellular vesicles (sEVs) secreted by adipocytes (Adipo-sEVs) or adipose tissue are abundant in nicotinamide phosphoribosyltransferase (NAMPT), which is the key NAD+ biosynthetic enzyme in mammals. Systemic injection of these sEVs significantly improves physical activity and extends the lifespan of aged mice by increasing NAD+ levels. However, to date, the therapeutic potential of Adipo-sEVs in other age-associated disease models, such as IVDD, has not been explored. In this study, we investigated the therapeutic effects of Adipo-sEVs on senescence of nucleus pulposus cells (NPCs) and cartilaginous endplate cells (EPCs). In vitro, Adipo-sEVs could rejuvenate the senescence of NPCs and EPCs. Age-related dysfunctions were also ameliorated by Adipo-sEVs by delivering NAMPT and activating NAD+ biosynthesis and the Sirt1 pathway. Further in vivo experiments revealed that Adipo-sEV-mediated delivery of NAMPT attenuated IVDD in rats by rejuvenating senescent NPCs and EPCs. Collectively, the results indicate a new cell-free tool and provide a promising sEV-mediated delivery method of NAMPT as a therapeutic approach for IVDD clinically.

To investigate the effects of acupuncture on phosphorylated P38 mitogen-activated protein kinase (p-P38MAPK), intercellular adhesion molecule-1 (ICAM-1), interferon γ (IFN-γ), and eosinophilic granulocytes (EOS) in lung tissue of asthmatic rats, and to explore the mechanism of acupuncture in regulating the apoptosis of EOS.Clean-grade male Wistar rats were randomly divided into normal, model, dexamethasone and acupuncture groups, 8 rats in each group. The asthmatic model was established by intraperitoneal injection of mixture suspension (1 mL) of 10% ovalbumin and 10% Al(OH)_3+ normal saline, followed by inhalation of atomized 1% ovalbumin solution for 30 min, once daily for 2 weeks to trigger occurrence of asthmatic symptoms. The rats in dexamethasone group were intraperitoneally injected with 0.9 mg/kg dexamethasone since day 15 once a day for two consecutive weeks. In the acupuncture group, bilateral "Feishu" (BL13), "Pishu" (BL20), "Shenshu" (BL23), "Dingchuan" (EX-B1), and "Danzhong" (CV17) were selected for acupuncture treatment once every other day since day 15 for two consecutive weeks. Uniform reinforcing and reducing manipulation was carried out, and the needles were retained for 30 min. The pathological changes of the lung tissue were observed by hematoxylin-eosin (HE) staining. The apoptosis of EOS in the lung tissue of rats was detected by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The expression of p-P38MAPK in the lung tissue was detected by Western blot. The protein and mRNA levels of ICAM-1 and IFN-γ in the lung tissue were determined by immunohistochemistry and real-time PCR, respectively.The results of HE staining showed that the pulmonary alveoli and surrounding tissues were intact with no inflammatory cell infiltration in the normal group. The model group showed massive exudation of inflammatory materials and thickened pulmonary interstitium. The dexamethasone group and acupuncture group showed less damage of the alveolar structure and only a small number of inflammatory cells around the airway. Compared with the normal group, the apoptosis rate of EOS in lung tissue was decreased (Acupuncture may inhibit the P38MAPK signaling pathway, down-regulate ICAM-1 expression, and up-regulate IFN-γ expression to promote the apoptosis of EOS and reduce EOS aggregation, thus alleviating the inflammatory response of airway in asthma.

The loss of normal alveolar capillary and deregulated angiogenesis occurs simultaneously in idiopathic pulmonary fibrosis (IPF); however the contributions of specific endothelial subpopulations in the development of pulmonary fibrosis are poorly understood. Herein, we perform single-cell RNA sequencing to characterize the heterogeneity of endothelial cells (ECs) in bleomycin (BLM)-induced lung fibrosis in rats. One subpopulation, characterized by the expression of Nos3 and Cav1, is mostly distributed in non-fibrotic lungs and also highly expresses genes related to the “response to mechanical stimulus” and “lung/heart morphogenesis” processes. Another subpopulation of ECs expanded in BLM-treated lungs, characterized by Cxcl12, is observed to be closely related to the pro-fibrotic process in the transcriptome data, such as “regulation of angiogenesis,” “collagen binding,” and “chemokine activity,” and spatially localized to BLM-induced neovascularization. Using CellPhoneDB software, we generated a complex cell–cell interaction network, which predicts the potential roles of EC subpopulations in recruiting monocytes, inducing the proliferation of fibroblasts and promoting the production and remolding of the extracellular matrix (ECM). Taken together, our data demonstrate the high degree of heterogeneity of ECs in fibrotic lung and it is proposed that the interaction between ECs, macrophages, and stromal cells contributes to pathologic IPF. Supplementary Information The online version contains supplementary material available at 10.1007/s10456-021-09795-5.

The purpose of the present study was to investigate the effects of drinking water alkaline mineral complex (AMC) supplementation on growth performance, intestinal morphology, inflammatory response, immunity, antioxidant defense system, and barrier functions in weaned piglets. In a 15-d trial, 240 weaned piglets (9.35 ± 0.86 kg) at 28 d of age (large white × landrace × Duroc) were randomly divided into two groups: the control (Con) group and the AMC group. Drinking water AMC supplementation improved (P0.01) final body weight (BW) and average daily gain (ADG) in weaned piglets compared to the Con group. Importantly, AMC reduced (P0.01) the feed-to-gain (F:G) ratio. AMC water improved the physical health conditions of piglets under weaning stress, as reflected by the decreased (P0.05) hair score and conjunctival score. Moreover, there was no significant (P0.05) difference in relatively small intestinal length, organ (liver, spleen, and kidney) indices, or gastrointestinal pH value in weaned piglets between the two groups. Of note, AMC significantly promoted the microvilli numbers in the small intestine and effectively ameliorated the gut morphology damage induced by weaning stress, as evidenced by the increased (P0.05) villous height (VH) and ratio of VH to crypt depth. Additionally, AMC lessened the levels of lipopolysaccharide (LPS, P0.01) and the contents of IL1β (P0.05), and TNF-α (P0.05) in the weaned piglet small intestine. Conversely, the gut immune barrier marker, secretory immunoglobulin A (sIgA) levels in serum and small intestine mucosa were elevated after AMC water treatment (P0.01). Furthermore, AMC elevated the antioxidant mRNA levels of (P0.05) SOD 1-2, (P0.01) CAT, and (P0.01) GPX 1-2 in the small intestine. Likewise, the mRNA levels of the small intestine tight junction factors Occludin (P0.01), ZO-1 (P0.05), Claudin 2 (P0.01), and Claudin 5 (P0.01) in the AMC treatment group were notably higher than those in the Con group. In conclusion, drinking water AMC supplementation has an accelerative effect on growth performance by elevating gut health by improving intestinal morphology, the inflammatory response, the antioxidant defense system, and barrier function in weaned piglets.The piglet suffers vital physiological, environmental, and social challenges when it is weaned from the sow that can predispose the piglet to subsequent diseases and other production losses, and these challenges are responsible for serious economic losses to the swine industry. Weaning stress induces intestinal injury, decreased immunity, and digestive system dysfunction, which then reduces feed intake and inhibits the growth performance of piglets. It is well known that alternatives to antibiotics for preventing weaning stress in weaned farm animals are sorely needed. The biologically beneficial effects of alkaline mineral water are widely reported. Alkaline mineral complex (AMC), as an immunomodulator, is considered to have antistress effects in the swine industry. In addition, treatment through drinking water is considered to be an efficient and low-cost feasible disease control strategy. Drinking water AMC supplementation is expected to exert health benefits in pigs; however, the responses of weaned piglets to water supplemented with AMC have not been fully explored. Thus, this study explored the effects of drinking water AMC supplementation on growth performance and gut health in weaned piglets. Our results showed that AMC water supplementation conspicuously enhanced the growth performance by improving the gut health.

In the modern poultry industry, newly hatched chicks are unavoidably transported from the hatching to the rearing foster. Stress caused by multiple physical and psychological stressors during transportation is particularly harmful to the liver. Astragalus polysaccharide (APS) possesses multiple benefits against hepatic metabolic disorders. Given that transport stress could disturb hepatic glucolipid metabolism and the role of APS in metabolic regulation, we speculated that APS could antagonize transport stress-induced disorder of hepatic glucolipid metabolism. Firstly, newly hatched chicks were transported for 0, 2, 4, and 8 h, respectively. Subsequently, to further investigate the effects of APS on transport stress-induced hepatic glucolipid metabolism disturbance, chicks were pretreated with water or APS and then subjected to transport treatment. Our study suggested that APS could relieve transport stress-induced lipid deposition in liver. Meanwhile, transport stress also induced disturbances in glucose metabolism, reflected by augmented mRNA expression of key molecules in gluconeogenesis and glycogenolysis. Surprisingly, APS could simultaneously alleviate these alterations via peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α)/Sirtuin 1 (SIRT1)/AMP-activated protein kinase (AMPK) pathway. Moreover, APS treatment regulated the level of peroxisome proliferator-activated receptor alpha (PPARα) and peroxisome proliferator-activated receptor gamma (PPARγ), thereby alleviating transport stress-induced alterations of VLDL synthesis, cholesterol metabolism, lipid oxidation, synthesis, and transport-related molecules. These findings indicated that APS could prevent the potential against transport stress-induced hepatic glucolipid metabolism disorders via PGC-1α/SIRT1/AMPK/PPARα/PPARγ signaling system.In the modern poultry industry, newly hatched chicks are unavoidably transported from the hatching to the rearing foster. During transportation, chicks are frequently subjected to various physical and psychological stressors, which can lead to alterations in blood composition, hormones, metabolites, enzymes, and behavior. These alterations adversely affect animal health and welfare. Stress caused by transportation is especially harmful to liver, which can cause significant effects on liver function, and disturb hepatic lipid metabolism and glucose metabolic. The current study demonstrated that Astragalus polysaccharide (APS) possesses multiple benefits against hepatic metabolic disorders. Administration of APS to chicks before transport could prevent transport-induced stress and hepatic glucolipid metabolism disorders.

To observe the effect of acupoint catgut embedment(ACE) on expression of p38 mitogen activated protein kinase (p38MAPK), intercellular adhesion molecule-1(ICAM-1), interleukin-4 (IL-4) and eosinophils (EOS) in lung tissue of asthmatic rats, so as to explore its mechanism underlying improvement of asthma.Forty male Wistar rats were randomly divided into control, model, ACE and dexamethasone (DEX) groups, with 10 rats in each group. The asthmatic model was established by intraperitoneal injection of mixture suspension (1 mL) of ovalbumin (OVA,10%) and 10% Al (OH)Findings of optical microscope and TEM showed obvious bronchial deformation and inflammatory cell infiltration, rupture of EOS cell membrane, uneven cytoplasm with swelling and uneven density of eosinophilic granules in EOS of the model group, which was relatively milder in the ACE and DEX groups. Compared with the control group, the EOS number in BALF and the expressions of p38MAPK, ICAM-1 and IL-4 mRNAs in the lung tissue were significantly increased (Catgut embedding at acupoints may alleviate the airway inflammatory response in asthma rats, which may be related with its effects in down-regulating p38MAPK signaling, ICAM-1 and IL-4 mRNA expression, reducing the aggregation of EOS, and promoting the apoptosis of EOS.

Inositol 1, 4, 5-trisphosphate (IP3) signaling-mediated calcium release drives the contraction of vascular smooth muscles and hence regulates blood vessel volume and blood pressure. Melatonin supplementation has been suggested to be beneficial for hypertension. To determine whether the blood pressure-lowering effect of melatonin was accounted for by IP3 signaling, we evaluated the vasoconstriction response and IP3 signaling in isolated mouse thoracic aortic rings during melatonin incubation. C57BL/6 mice were given intraperitoneal injections daily with melatonin, and the systolic blood pressure and contractility of aortic rings from melatonin-treated mice were decreased, and the contraction suppression effect of melatonin was attributed to the impaired expression of contractile proteins in vascular smooth muscle cells rather than IP3 signaling. Our results further showed that melatonin increased the expression of γ-secretase, which could cleave and release the notch intracellular domain, and the notch intracellular domain prevented the transcription of contractile genes by interfering with the interaction between serum response factor and myocardin, the master regulator of contractile protein. In this article, we report a novel mechanism by which melatonin regulates smooth muscle contractility that does not depend on IP3 signaling.

Background and Purpose: RAS mutations limit the effectiveness of anti-epidermal growth factor receptor (EGFR) monoclonal antibodies in combination with chemotherapy for metastatic colorectal cancer (mCRC) patients. Therefore, new cell death forms have focused on identifying indirect targets to inhibit Ras-induced oncogenesis. Recently, emerging evidence has shown the potential of triggering ferroptosis for cancer therapy, particularly for eradicating aggressive malignancies that are resistant to traditional therapies. Methods: KRAS mutant CRC cell HCT116 and Lovo were treated with cetuximab and β-elemene, a bioactive compound isolated from Chinese herb Curcumae Rhizoma. Ferroptosis and epithelial-mesenchymal transformation (EMT) were detected in vitro and in vivo. Orthotopic CRC animal model were established and the tumor growth was monitored by IVIS bioluminescence imaging. Tumor tissues were collected to determine ferroptosis effect and the expression of EMT markers after the treatment. Results: CCK-8 assay showed that synergetic effect was obtained when 125 µg/ml β-elemene was combined with 25 µg/ml cetuximab in KRAS mutant CRC cells. AV/PI staining suggested a non-apoptotic mode of cell death after the treatment with β-elemene and cetuximab. In vitro, β-elemene in combination with cetuximab was shown to induce iron-dependent reactive oxygen species (ROS) accumulation, glutathione (GSH) depletion, lipid peroxidation, upregulation of HO-1 and transferrin, and downregulation of negative regulatory proteins for ferroptosis (GPX4, SLC7A11, FTH1, glutaminase, and SLC40A1) in KRAS mutant CRC cells. Meanwhile, combinative treatment of β-elemene and cetuximab inhibited cell migration and decreased the expression of mesenchymal markers (Vimentin, N-cadherin, Slug, Snail and MMP-9), but promoted the expression of epithelial marker E-cadherin. Moreover, ferroptosis inhibitors but not other cell death suppressors abrogated the effect of β-elemene in combination with cetuximab on KRAS mutant CRC cells. In vivo, co-treatment with β-elemene and cetuximab inhibited KRAS mutant tumor growth and lymph nodes metastases. Conclusions: Our data for the first time suggest that the natural product β-elemene is a new ferroptosis inducer and combinative treatment of β-elemene and cetuximab is sensitive to KRAS mutant CRC cells by inducing ferroptosis and inhibiting EMT, which will hopefully provide a prospective strategy for CRC patients with RAS mutations.

The tripartite motif containing 23 (TRIM23) gene is a member of the tripartite motif (TRIM) family that participates in many pathophysiological processes. However, the role of TRIM23 in lung adenocarcinoma (LUAD) remains unclear. In the present study, TRIM23 was first screened by next‐generation sequencing between the cisplatin (DDP)‐resistant A549/DDP cell line and the parental A549 cell line, combined with integrated analysis of the Gene Expression Omnibus (GEO) data (E‐GEOD‐43493 and E‐GEOD‐43494). The expression of TRIM23 was then verified to be upregulated in the DDP‐resistant LUAD cells and tissues. The knockdown of TRIM23 expression in A549/DDP cells caused increased apoptosis, decreased IC50 values of DDP, NF‐κB nuclear translocation, inhibition of cell proliferation in vitro and in vivo, inhibition of GLUT1/3 expression, glucose uptake, and lactate and ATP production. TRIM23 overexpression resulted in the opposite effects in A549 cells. In addition, the inhibition of proliferation in A549 cells caused by NF‐κB signaling inhibitor PTDC or glycolysis inhibitor 3‐BrPA could be weakened by TRIM23 overexpression. Furthermore, immunohistochemical analysis revealed that TRIM23 was upregulated in 46.1% (70/152) of LUAD cases, and elevated TRIM23 expression was correlated with high expression of NF‐κB, poor cellular differentiation, and adverse overall survival (OS) and disease‐free survival (DFS). In conclusion, our study demonstrates that TRIM23 acts as an oncogene in LUAD and promotes DDP resistance by regulating glucose metabolism via the TRIM23/NF‐κB/ GLUT1/3 axis., Our study demonstrates that TRIM23 acts as an oncogene in lung adenocarcinoma and promotes DDP resistance by regulating glucose metabolism via the TRIM23/NF‐κB/GLUT1/3 axis.

Hepatocellular carcinoma (HCC) is the major cause of liver cancer-associated morality. Metformin, used for treating type 2 diabetes, has antitumor activity and reduces the risk of some diabetes-related tumors, such as liver and breast cancer. However, the mechanisms underlying metformin's effects in HCC remain unclear. To identify genes associated with metformin treatment in HCC, we conducted transcriptomic and proteomic analyses in HCC cells treated with or without metformin. We identified 41 differentially expressed genes upon metformin treatment. Among them, Ataxin 7 Like 3B (ATXN7L3B), which is a negative regulator of the Spt-Ada-Gcn5 acetyltransferase (SAGA) deubiquitinase (DUB) module and has relatively unknown functions in cancer, attracted our attention. We observed that metformin reduced ATXN7L3B level in HCC cells. ATXN7L3B expression was significantly negatively correlated with survival in liver cancer patients. We also demonstrated that ATXN7L3B promoted HCC stemness. Metformin treatment decreased ATXN7L3B-induced tumor-initiating ability in a HCC mouse model, implying that metformin may inhibit cancer stemness by downregulating ATXN7L3B. Our study supports the antitumor activity of metformin and its potential as an anticancer drug for HCC treatment.

Objective: To establish an acquired aplastic anemia animal model for investigating the function of T lymphocyte and the pathogenesis and treatment of aplastic anemia(AA).To establish the acquired aplastic anemia mouse model through the X-ray irradiation in combination with lymphocytes injection. AA Group: the purified Pan T lymphocytes from the spleen of C57BL/6J mice were enriched and injected to the mice through tail vein(5×10Compared with 4, 5 Gy irradiated mice in AA groups, the survival time of 3 Gy irradiated AA groups was significantly prolonged. 3, 4 and 5 Gy X-ray irradiation combined with Pan T lymphocyte injection could successfully induced severe reduction of red blood cells, blood neutrophils, and platelets, severe reduction of bone marrow nucleated cells, severe bone marrow hematopoietic failure, and the significant expansion of T lymphocytes ratio in the bone marrow. CD43, 4 and 5 Gy X-ray irradiation combined with 5×10泛T淋巴细胞结合照射诱导的再生障碍性贫血小鼠模型的优化.建立适用于研究T淋巴细胞功能的获得性再生障碍性贫血(acquired aplastic anemia，AA)动物模型以及应用于再生障碍性贫血发病机制及治疗研究的动物模型.对X射线联合淋巴细胞注射诱导的获得性再生障碍性贫血小鼠模型进行优化。AA组：富集C57BL/6J小鼠的脾脏泛T细胞,按细胞数5×10相较于4、5 Gy照射的AA组，3 Gy照射的AA组生存期明显延长；经3、4、5 Gy X射线照射联合泛T淋巴细胞注射均可成功诱导受体鼠外周血红细胞、白细胞、血小板数严重减少，骨髓有核细胞严重减少，骨髓造血衰竭，骨髓中T淋巴细胞占比明显增加，CD43、4和5 Gy X射线的照射联合5×10

To observe the effect of proximal and distal acupoint catgut-embedding on the contents of prostaglandin EFemale Wistar rats were randomly divided into control, model, proximal catgut-embedding and distal catgut-embedding groups, with 8 rats in each group. The PD model was established by subcutaneous injection of estradiol ben-zoate (0.5 mg/rat on the first day and 10th day, and 0.2 mg/rat from the 2nd to the 9th day) and intraperitoneal injection of oxytocin (2 U/rat, on the 11th day). Catgut-embedding was applied at bilateral "Guanyuan"(CV4) and "Ciliao"(BL32) in the proximal catgut-embedding group, and bilateral "Sanyinjiao" (SP6) and "Diji"(SP8) in the distal catgut-embedding group. After the treatment, the body writhing times in 30 min and uterine mass were recorded, PGE2 and PGF2αin uterus and IL-2 in serum were assayed by using ELISA, the activity of NK cell in the spleen was detected using MTT colorimetry.Following modeling, the body writhing times of the model group was increased than that of the control group (Both proximal and distal acupoint catgut-embeding can increase PGE

Background:Fatty infiltration, inflammation, and apoptosis are common degenerative changes in patients with chronic rotator cuff tears that can lead to muscle atrophy and can even result in massive irreparable rotator cuff tears. Some data have demonstrated the proregenerative, anti-inflammatory, and anti-apoptotic properties of stem cell–derived exosomes in some orthopaedic disorders, but their effect on torn rotator cuff muscles has never been investigated.Purpose:To study the effect of exosomes isolated from human adipose-derived stem cells (ASCs-Exos) on muscle degeneration, regeneration, and biomechanical properties in a rat model of a massive rotator cuff tear (MRCT).Study Design:Controlled laboratory study.Methods:A bilateral supraspinatus and infraspinatus tenotomy was performed on rats to create an MRCT model. Forty-two rats were randomly assigned to 3 groups: the sham surgery group, the saline group (lesions treated with a saline injection), and the ASCs-Exos group (lesions treated with an ASCs-Exos injection). Wet muscle weight, fatty infiltration, inflammation, vascularization, regeneration, and biomechanical properties were evaluated at 8 and 16 weeks after surgery.Results:The results revealed that the ASCs-Exos treatment could prevent the atrophy, fatty infiltration, inflammation, and vascularization of muscles in the MRCT model ( P < .001). Additionally, the myofiber regeneration and biomechanical properties of ASCs-Exos-treated rotator cuffs were significantly elevated compared with those in the saline-treated group ( P < .001).Conclusion:This study demonstrates that ASCs-Exos can effectively decrease atrophy and degeneration and improve muscle regeneration and biomechanical properties in torn rotator cuff muscles.Clinical Relevance:ASCs-Exos can be used as a new cell-free approach to prevent the muscle degeneration associated with torn rotator cuffs and may be helpful to repair torn rotator cuffs. Nevertheless, further work needs to be done in a large animal model owing to the inherent regenerative potential possessed by rodents.

Background Angiogenesis, as an endogenous repair mechanism, plays crucial roles in wound healing and tissue regeneration. However, this process is impaired in the elderly due to aging-related vascular endothelial dysfunction. This study was aimed to explore the pro-angiogenic effects of exosomes from human embryonic stem cells (ESC-Exos) in aged mice of pressure-induced ulcer model and the underlying mechanism. Methods Pressure ulcer wounds were created on the back of d-galactose-induced aging mice. ESC-Exos were locally applied onto the wound beds, with PBS as control. The effects of ESC-Exos on wound healing were analyzed by measuring wound closure rates, histological and immunofluorescence analyses. Then, the anti-aging effect of ESC-Exos on vascular endothelial cells was tested in an in vitro d-galactose-induced HUVEC senescence model. Results ESC-Exos could accelerate wound closure and enhance angiogenesis, and the senescence of vascular endothelial cells was significantly ameliorated after ESC-Exos treatment. In vitro, ESC-Exos could rejuvenate the senescence of endothelial cells and recover compromised proliferation, migratory capacity, and tube formation. This recovery was Nrf2-activation-dependent, since cotreatment with Nrf2 inhibitor Brusatol could abolish the rejuvenative effects of ESC-Exos. Further study revealed that miR-200a was highly enriched in ESC-Exos and played a crucial role in ESC-Exos-mediated rejuvenation through downregulating Keap1, which negatively regulates Nrf2 expression. Conclusions ESC-Exos ameliorate endothelial senescence by activating Nrf2 and recover aging-related angiogenic dysfunction, thereby accelerating wound healing in aged mice. ESC-Exos might be a natural nano-biomaterial for aging-related diseases therapy. Electronic supplementary material The online version of this article (10.1186/s13287-019-1253-6) contains supplementary material, which is available to authorized users.

This study focuses on investigating the role of interleukin‐1β (IL‐1β) in functional regeneration following nerve injury in mice. A microarray‐based mRNA profiling study was used to analyze the expression level of IL‐1β in peripheral nerve regeneration. Quantitative real‐time polymerase chain reaction and Western blot were applied to assess the IL‐1β expressions of C57BL/6J‐crush and C57BL/6J‐crush+IL‐1β mice at different post‐injury time‐points after the standard sciatic nerve crush injury. The outcomes of nerve regeneration were evaluated by behavioral tests. IL‐1β was found to be up‐regulated in peripheral nerve regeneration and significantly raised on the 3rd day and returned to normal levels on the 14th day after nerve injury. Compared with C57BL/6J‐crush+IL‐1β mice, the nerve regeneration of C57BL/6J‐crush mice was worse after nerve crush injury. IL‐1β increased mechanical sensitivity and stimulated amplitude. IL‐1β could benefit the recovery of sciatic nerve crush injury by facilitating nerve regeneration.

To explore the efficacy difference between electroacupuncture (EA) at "Zusanli" (ST36) and "Baihui" (GV20) for inflammatory pain and cerebral ischemia-reperfusion injury (CIRI) in rats.In 1st part of this study, 90 male SD rats were randomly divided into sham-operation, model (induced by occlusion of the middle cerebral artery and reperfusion), GV20 EA, ST36 EA，and sham EA groups ((1) Twenty-four hours following CIRI, the NDS and infarct volume were significantly increased in the model group compared with the sham-operation group (Our data do not support the specificity of functions at least between GV20 EA and ST36 EA in both CIRI and inflammatory pain model rats. This is the first study reporting the effect of EA at GV20 for relieving CFA-induced inflammatory pain.

Background Advances in regenerative medicine technologies have been strongly proposed in the management of thyroid diseases. Mechanistically, the adoption of thyroid bioengineering requires a scaffold that shares a similar three-dimensional (3D) space structure, biomechanical properties, protein component, and cytokines to the native extracellular matrix (ECM). Methods 24 male New Zealand white rabbits were used in this experimental study. The rabbit thyroid glands were decellularized by immersion/agitation decellularization protocol. The 3D thyroid decellularization scaffolds were tested with histological and immunostaining analyses, scanning electron microscopy, DNA quantification, mechanical properties test, cytokine assay and cytotoxicity assays. Meanwhile, the decellularization scaffold were seeded with human thyroid follicular cells, cell proliferation and thyroid peroxidase were determined to explore the biocompatibility in vitro. Results Notably, through the imaging studies, it was distinctly evident that our protocol intervention minimized cellular materials and maintained the 3D spatial structure, biomechanical properties, ECM composition, and biologic cytokine. Consequently, the decellularization scaffold was seeded with human thyroid follicular cells, thus strongly revealing its potential in reinforcing cell adhesion, proliferation, and preserve important protein expression. Conclusions The adoption of our protocol to generate a decellularized thyroid scaffold can potentially be utilized in transplantation to manage thyroid diseases through thyroid bioengineering.

Ferroptosis, a novel form of programmed cell death, is characterized by iron-dependent lipid peroxidation and has been shown to be involved in multiple diseases, including cancer. Stimulating ferroptosis in cancer cells may be a potential strategy for cancer therapy. Therefore, ferroptosis-inducing drugs are attracting more attention for cancer treatment. Here, we showed that erianin, a natural product isolated from Dendrobium chrysotoxum Lindl, exerted its anticancer activity by inducing cell death and inhibiting cell migration in lung cancer cells. Subsequently, we demonstrated for the first time that erianin induced ferroptotic cell death in lung cancer cells, which was accompanied by ROS accumulation, lipid peroxidation, and GSH depletion. The ferroptosis inhibitors Fer-1 and Lip-1 but not Z-VAD-FMK, CQ, or necrostatin-1 rescued erianin-induced cell death, indicating that ferroptosis contributed to erianin-induced cell death. Furthermore, we demonstrated that Ca2+/CaM signaling was a critical mediator of erianin-induced ferroptosis and that blockade of this signaling significantly rescued cell death induced by erianin treatment by suppressing ferroptosis. Taken together, our data suggest that the natural product erianin exerts its anticancer effects by inducing Ca2+/CaM-dependent ferroptosis and inhibiting cell migration, and erianin will hopefully serve as a prospective compound for lung cancer treatment.

The aim of this study was to explore the roles of GPX2, a member of the glutathione peroxidase family (GPXs, GSH-Px), in cisplatin (DDP) resistance in lung adenocarcinoma (LUAD). GPX2 was found to be the most significantly upregulated gene in a DDP-resistant A549/DDP cell line compared with the parental A549 cell line by RNA sequencing. The knockdown of GPX2 expression in A549/DDP cells inhibited cell proliferation in vitro and in vivo, decreased the IC50 values of DDP, induced apoptosis, inhibited the activities of GSH-Px and superoxide dismutase (SOD), inhibited ATP production and glucose uptake, and increased malondialdehyde (MDA) and reactive oxygen species (ROS) production; while GPX2 overexpression in A549 cells resulted in the opposite effects. Using gene set enrichment analysis (GSEA), we found that GPX2 may be involved in DDP resistance through mediating drug metabolism, the cell cycle, DNA repair and energy metabolism, and the regulation of an ATP-binding cassette (ABC) transporters member ABCB6, which is one of the hallmark genes in glycolysis. Moreover, immunohistochemistry revealed that GPX2 was upregulated in 58.6% (89/152) of LUAD cases, and elevated GPX2 expression was correlated with high expression of ABCB6, high 18-fluorodeoxyglucose (18F-FDG) uptake, and adverse disease-free survival (DFS) in our cohort. The Cancer Genome Atlas (TCGA) data also indicated that GPX2 expression was higher in LUAD than it was in normal lung tissues, and the mRNA expression levels of GPX2 and ABCB6 were positively correlated. In conclusion, our study demonstrates that GPX2 acts as oncogene in LUAD and promotes DDP resistance by regulating oxidative stress and energy metabolism.

Mounting evidence suggests that an emerging G protein-coupled receptor, TGR5, plays a crucial role ranging from metabolic diseases to cancers. However, the biological functions of TGR5 in non-small cell lung cancer (NSCLC) remain elusive. We found that TGR5 was aberrantly expressed in NSCLC and positively correlated with an advanced clinical stage in NSCLC patients. We further discovered that TGR5 knockdown prevented JAK2 and STAT3 phosphorylation and repressed the expression of STAT3 target genes, thus inhibiting cell proliferation, migration and invasion in NSCLC. Moreover, the promotive effects of TGR5 were significantly reversed by a JAK2 inhibitor or STAT3 knockdown. Additionally, we demonstrated a positive correlation between TGR5 and p-STAT3 expression in NSCLC tissue samples. Patients with both high TGR5 and p-STAT3 expression had the worst prognosis. In addition, the serum levels of deoxycholic acid (DCA), ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) in NSCLC patients were much higher than those in the healthy controls and the patients with higher serum DCA levels had stronger TGR5 expression simultaneously. Moreover, DCA markedly promoted NSCLC cell migration and invasion through a TGR5-dependent way. Taken together, these results indicate that TGR5 drives cell growth and migration through JAK2/STAT3 signaling pathway in NSCLC.

Summary Stem cells, with their therapeutic potential in tissue repair and regeneration, have been widely used in translational medicine. Recent evidence suggests that the beneficial effects are mediated largely by their paracrine actions rather than the engraftment and differentiation at the injured sites. Extracellular vesicles (EVs), actively released from cells, play important roles in cell-to-cell communication and display multiple functions in tissue regeneration. In the present report, we will briefly review the current knowledge related to the therapeutic potential of EVs, particularly stem cell or progenitor cell-derived ones for promoting tissue repair and regeneration, and focus on the restorative properties of exosomes/microvesicles in cutaneous wound healing, bone regeneration, hindlimb ischemia, and vascular injury repair.

Gastroesophageal reflux (GER) and microaspiration of duodenogastric refluxate have been recognized as a risk factor for pulmonary fibrosis. Recent evidence suggests that bile acid microaspiration may contribute to the development of lung fibrosis. However, the molecular evidence is scarce and the underlying mechanisms remain to be elucidated. We have recently demonstrated that bile acids induce activation of alveolar epithelial cells (AECs) and lung fibroblasts in vitro . In the present study, a rat model of bile acid microaspiration was established by weekly intratracheal instillation of three major bile acids including chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and lithocholic acid (LCA). Repeated microaspiration of CDCA, DCA, and LCA caused fibrotic changes, including alveolar wall thickening and extensive collagen deposition, in rat lungs. Bile acid microaspiration also induced alveolar epithelial–mesenchymal transition (EMT), as indicated by up-regulation of mesenchymal markers α-smooth muscle actin (α-SMA) and vimentin, as well as down-regulaton of epithelial markers E-cadherin and cytokeratin in alveolar epithelium of rat lungs. The expression of fibrogenic mediators, including transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and periostin, was significantly elevated in rat lungs exposed to microaspiration of bile acids. Furthermore, microaspiration of bile acids also induced p-Smad3 and farnesoid X receptor (FXR) expression in rat lungs. Our findings suggest that microaspiration of bile acids could promote the development of pulmonary fibrosis in vivo , possibly via stimulating fibrogenic mediator expression and activating TGF-β1/Smad3 signaling and FXR. * α-SMA, : α-smooth muscle actin; AEC, : alveolar epithelial cell; BALF, : bronchoalveolar lavage fluid; bFGF, : basic fibroblast growth factor; BOS, : bronchiolitis obliterans syndrome; CDCA, : chenodeoxycholic acid; CTGF, : connective tissue growth factor; DCA, : deoxycholic acid; ECM, : excessive extracellular matrix; EMT, : epithelial–mesenchymal transition; FXR, : farnesoid X receptor; GER, : gastroesophageal reflux; H&E, : hematoxylin and eosin; HRP, : horseradish peroxidase; IHC, : immunohistochemical; IOD, : integrated optical density; IPF, : idiopathic pulmonary fibrosis; LCA, : lithocholic acid; OB, : obliterative bronchiolitis; TGF-β1, : transforming growth factor-β1; VEGF, : vascular endothelial growth factor

To observe the effect of herbal-cake-partitioned moxibustion (HCPM) of "Shenque" (CV8) and "Daheng" (SP15) on abdominal pain, plasma β-endorphin (β-EP), uterine prostaglandin EA total of 40 female rats were randomized into blank control, model, CV8-direct moxibustion, CV8-HCPM and SP15-HCPM groups (Compared with the control group, the writhing times and the content of PGFMoxibustion of both CV8 and SP15 can relieve abdominal pain in PD rats, which may be closely associated with its effect in suppressing PD-induced decrease of plasma β-EP and uterine PGE

D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS1000) is the most active water-soluble derivative of vitamin E and has been widely used as a carrier of solvents, plasticizers, emulsifiers, absorbent agents and refractory drug delivery systems. However, its anti-hepatocellular carcinoma (HCC) properties have not been explored. HCC cells were treated with different concentrations of TPGS1000. Cell survival was tested by CCK8 assay, and cell migration was tested by wound healing and Transwell assay. EdU staining verified cell proliferation, and signalling pathway was assayed by Western blot analysis. The BALB/c-nu mouse xenograft model was established to test HCC cell growth in vivo. In vitro TPGS1000 significantly inhibited the viability and mobility of HCC cells (HepG2, Hep3B and Huh7) in a dose-dependent manner. Cell cycle analysis indicated that TPGS1000 treatment arrested the HCC cell cycle in the G0/G1 phase, and induction of cell apoptosis was confirmed by TUNEL and Annexin V-7-AAD staining. Further pharmacological analysis indicated that collapse of the transmembrane potential of mitochondria, increased ROS generation, PARP-induced cell apoptosis and FoxM1-p21-mediated cell cycle arresting, were involved in the anti-HCC activity of TPGS1000. Moreover, treatment in vivo with TPGS1000 effectively impaired the growth of HCC xenografts in nude mice.

The different lineage-specific biological properties of articular cartilage and subchondral bone present a great challenge in the construction of bi-lineage scaffolds for simultaneous osteochondral regeneration. To overcome this challenge, strontium incorporated calcium silicate (Sr-CS) ceramic was prepared for bi-lineage formation of scaffolds in this study. The positive result of Sr-CS in the regeneration of osteochondral defects was first proven by its improved effect on the osteogenesis and chondrogenesis induction of mesenchymal stem cells (MSCs). After that, scaffold-mediated macrophage polarization between classically activated inflammatory macrophages (termed M1Ф) and alternatively activated inflammatory macrophages (termed M2Ф) was assayed to investigate whether the incorporation of Sr into calcium silicate could alter host-to-scaffold immune response. Furthermore, the interactions between Sr-CS pretreated macrophages and MSCs differentiation were performed to prove the enhancement effect of suppressed inflammatory response on osteogenesis and chondrogenesis. In vivo transplantation showed that the Sr-CS scaffolds distinctly improved the regeneration of cartilage and subchondral bone, as compared to the calcium silicate scaffolds. On the one hand, the mechanism attributes to enhancement of strontium on the osteogenic and chondrogenic differentiation of MSCs. On the other hand, the reason can partially be attributed to suppressed synovial inflammatory response, which has improved effects on enhancement of osteogenesis and chondrogenesis. These findings suggest that monophasic Sr-CS scaffolds with a bi-lineage conducive property and an inflammatory response regulatory property represents a viable strategy for simultaneous regeneration of osteochondral defects.

The exosomes are derived from mesenchymal stem cells (MSCs) and may be potentially used as an alternative for cell therapy, for treating diabetic wounds, and aid in angiogenesis. This study, aimed to investigate whether exosomes originated from bone marrow-derived MSCs (BMSCs) preconditioned by deferoxamine (DFO-Exos) exhibited superior proangiogenic property in wound repair and to explore the underlying mechanisms involved. Human umbilical vein endothelial cells (HUVECs) were used for assays involving cell proliferation, scratch wound healing, and tube formation. To test the effects in vivo, streptozotocin-induced diabetic rats were established. Two weeks after the procedure, histological analysis was used to measure wound-healing effects, and the neovascularization was evaluated as well. Our findings demonstrated that DFO-Exos activate the PI3K/AKT signaling pathway via miR-126 mediated PTEN downregulation to stimulate angiogenesis in vitro. This contributed to enhanced wound healing and angiogenesis in streptozotocin-induced diabetic rats in vivo. Our results suggest that, in cell-free therapies, exosomes derived from DFO preconditioned stem cells manifest increased proangiogenic ability.

17β-estradiol (E2) has been shown to have neuroprotective effects in different central nervous system diseases. The mechanisms underlying estrogen neuroprotection in spinal cord injury (SCI) remain unclear. Previous studies have shown that autophagy plays a crucial role in the course of nerve injury. In this study, we showed that E2 treatment improved the restoration of locomotor function and decreased the loss of motor neurons in SCI rats. Real-time PCR and western blot analysis revealed that the protective function of E2 was related to the suppression of LC3II and beclin-1 expression. Immunohistochemical study further confirmed that the immunoreactivity of LC3 in the motor neurons was down-regulated when treated with E2. In vitro studies demonstrated similar results that E2 pretreatment decreased the autophagic activity induced by rapamycin (autophagy sensitizer) and increased viability in a PC12 cell model. These results indicated that the neuroprotective effects of E2 in SCI are partly related to the suppression of excessive autophagy.

Background Asthma is closely associated with tobacco smoking (TS) and is more difficult to effectively treat after exposure to TS. Objective To observe the effects of TS on the expression of endothelin-2 (ET-2) and airway inflammation in asthmatic rats and to explore the related mechanisms. Methods We established an animal model of asthma with ovalbumin (OVA)/Al(OH)3 and subjected different animal groups to TS and/or dexamethasone/bosentan. The differences in the inflammatory cell infiltration, the pathological changes to the bronchial wall and the bronchial smooth muscle thickness, and the expression of ET-2, c-Jun amino terminal kinase (JNK1/2), malondialdehyde (MDA), and glutathione peroxidase (GSH) in the lung tissue and of interleukin (IL)-7 in bronchoalveolar lavage fluid (BALF) were assessed. Results Exposure to TS or OVA caused an obvious increase in the inflammatory cells in the BALF over what was observed in the control group. In asthma models, the expression of ET-1, JNK1/2, MDA, and GSH in the lung tissues, as well as that of IL-17 in the BALF, was increased. After treatment with dexamethasone/bosentan, the expression of IL-17, JNK1/2, MDA, and GSH decreased compared to the smoking group; airway inflammation and the staining intensity in the lung tissue were also reduced. Conclusion TS exposure can clearly exacerbate airway inflammation in asthmatic rats, while bosentan can alleviate airway inflammation through regulation of the ET-2/JNK1/2 signalling pathway.

To explore the underlying mechanism of acupoint catgut embedding in improving primary dysme-norrhea (PD) in rats based on functional activities of the neuro-endocrine-immune (NEI) network.Forty female rats were equally randomized into blank control, PD model, medication, and acupoint catgut embedding groups. The PD model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1The body writhing times were no-tably more in the model group than in the control group (The acupoint catgut embedding has a significant efficacy in relieving PD in rats, which may be related to its effect in up-regulating plasma β-EP, uterus PGE

The farnesoid X receptor (FXR) regulates inflammation and immune responses in a subset of immune-mediated diseases. We previously reported that FXR expression promotes tumor cell proliferation in non-small cell lung cancer (NSCLC). Here we study the relevance of FXR to the immune microenvironment of NSCLC. We found an inverse correlation between FXR and PD-L1 expression in a cohort of 408 NSCLC specimens; from this, we identified a subgroup of FXR

Previous studies have shown beneficial effects of mesenchymal stem cells (MSCs) transplantation in many autoimmune diseases. However, few studies have focused on the effects of MSCs on autoimmune hepatitis. In our study, we investigated the therapeutic effects of BMSCs (bone mesenchymal stem cells) transplantation in mouse experimental autoimmune hepatitis (EAH) and explored the potential mechanism. BMSCs were injected intravenously into EAH mice. Then, serum levels of ALT and AST, and pathologic alteration of liver tissue were measured to evaluate the liver function and inflammation degree. The expressions of programmed death ligand 1, IL-17 and IL-23 were detected by enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and western blotting. Upon serum biochemical levels and pathological examination, the BMSCs-treated mice especially with multiple dosing administration showed significantly reduction of liver damage. Moreover, the expression of IL-17 was down-regulated by BMSCs intervention as compared to the model group, whereas the PD-L1 and IL-23 were up-regulated following the administration of MSCs. In conclusion, the results of this study suggest that BMSCs transplantation, especially on multiple dosing, may exert immunosuppression effect to ameliorate EAH through the inhibition of IL-17 and up-regulation of PD-L1.

Objective To study the effect of cigarette smoke exposure on the expression of endothelin 2 (ET-2) in bronchial epithelium of asthmatic rats. Methods Asthma models were established through intraperitoneal injection of 1 mL chicken ovalbumin (OVA)/Al(OH)

The purpose of this study was to compare the effects of zinc oxide-eugenol (ZOE) and calcium hydroxide/iodoform paste (Vitapex), as root canal filling materials in pulpectomy, on delaying the root resorption of primary molars without permanent successors. Animal models without permanent successors were surgically established in beagle dogs. Root resorption was observed via periapical radiographs. The onset of root resorption of primary mandibular molars without successors occurred later (p

Paragonimiasis has previously been reported in the Southwest Provinces of China, including Chongqing and Sichuan. The construction of Three Gorges Dam, which was begun in 1994, has resulted in substantial changes to the depth and the flow pattern of the Yangtze River. To investigate epidemiology of the paragonimiasis, 724 people aged 2–49 years were selected and examined for Paragonimus infection by intradermal test (IDT), and enzyme-linked immunosorbent assay (ELISA). A survey of eating habits was conducted face-to-face in 5 endemic counties. Freshwater crabs were collected from seven sites in the 5 counties to determine the rate of infection with Paragonimus metacercariae. Additionally, we analyzed 213 case reports from the region dated 2006 to 2009. According to the survey results, the prevalence of Paragonimus skrjabini infection in freshwater crab was 39.65%, and the human infection rates detected by IDT and ELISA were 14.36% (104/724) and 7.46% (54/724), respectively. These results show that the Three Gorges Reservoir is still an area of high paragonimiasis endemicity. The water level of the Yangtze River has risen to 175 m, and the area of water coverage in the region has increased to 1084 km2. These conditions will favor the growth and expansion of the freshwater crab population.

Metastasis accounts for the majority of cancer-related deaths. Transforming growth factor β (TGF-β) is believed to promote late-stage cancer progression and metastasis by inducing epithelial-mesenchymal transition (EMT). We previously reported that MS80, a novel oligosaccharide sulfate, inhibits TGF-β1-induced pulmonary fibrosis by binding TGF-β1. In our study MS80 effectively inhibited TGF-β/Smad signaling in lung cancer cells, breast cancer cells, and model cell lines. In addition, MS80 inhibited TGF-β1-induced EMT, motility, and invasion in vitro. Moreover, MS80 significantly inhibited lung metastasis in orthotopic 4T1 xenografts. Notably, the MS80 treatment significantly increased the infiltration of CD8(+) T cells and decreased the infiltration of regulatory T cells in primary tumors and spleens in mice bearing 4T1 xenografts. Therefore, MS80 is a novel and promising candidate for treating metastatic malignancies by targeting TGF-β1-induced EMT and mediating immunosuppression.

Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3. Hypoxia-inducible factor-1-alpha (HIF-1-alpha) is a transcription factor specifically activated by hypoxia. 2-methoxyestradiol (2ME2) is an estradiol derivative and a known HIF-1-alpha inhibitor. 2ME2 decreased apoptosis by inhibiting HIF-1-alpha. The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of 2ME2. Male adult rats were used to make the global ischemia (GI) model. Ten minutes after GI, 2ME2 was injected intraperitoneally (16 mg/kg weight). Rats were killed at 6 hours, 12 hours, 24 hours, 48 hours, 96 hours, and 7 days. GI produced a marked increase in HIF-1-alpha expressions in the hippocampus at 6 hours and peaked at 48-96 hours. The expressions of survivin and caspase-3 were increased lightly in a similar time course. These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions. 2ME2 treatment reduced the expression of HIF-1-alpha, increased survivin expression, and decreased the expression of caspase-3. These results indicate that survivin and HIF-1-alpha were involved in the anti-apoptotic effect of 2ME2 treated following GI. 2ME2 may decrease the HIF-1-alpha expression, up-regulate the survivin expression, inhibit the expression of caspase-3, and finally reduce apoptosis after GI.

3'-Methoxypuerarin (3'-MOP) is an isoflavone extracted from radix puerariae. The aim of this study was to investigate the role and the mechanism of 3'-MOP in the protection of hippocampal neurons against cerebral ischemia/reperfusion (I/R) injury in rats. I/R injury was induced by a modified four-vessel occlusion model. Rats were randomly divided into an I/R group, an I/R + 3'-MOP group and a control group. Histological changes in the neurons of the hippocampal CA1 region were observed with hematoxylin and eosine (H&E) staining. The apoptotic neurons in the hippocampal CA1 area were counted with the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. The results showed that compared with the I/R group, 3'-MOP increased the number of surviving neurons in the hippocampal CA1 region (P < 0.001) and markedly reduced the number of apoptotic pyramidal neurons (P < 0.001) after I/R injury. In conclusion, 3'-MOP can protect hippocampal neurons against I/R injury by inhibiting apoptosis.

Amodiaquine (AQ) is routinely prescribed as an anti-malarial drug. Here, we evaluated AQ-induced toxicity in the male reproductive system. Eighty adult male Sprague-Dawley rats were randomly divided into four groups that received distilled water (control) or daily doses of 5 mg/kg body weight, 10 mg/kg, or 15 mg/kg AQ for 2 weeks. Testes morphology was analyzed using hematoxylin-and-eosin staining, terminal dUTP nicked-end labeling (TUNEL), and immunostaining whereas protein expression was determined by Western blotting. AQ dose-dependently led to abnormal spermatogenesis. Disruption of the blood-testis barrier and increased germ cell apoptosis were observed in all three AQ-treated groups. Interestingly, AQ-induced damage of spermatogenesis recovered over time, based on the survival of promyelocytic leukemia zinc-finger (PLZF)-positive, undifferentiated spermatogonia. Serum levels of luteinizing hormone and testosterone, as well as testicular testosterone levels, were not significantly altered in AQ-treated groups compared with controls. Collectively, our study suggests that AQ exerts substantial acute side effects on the reproductive systems of adult male rats by inducing the apoptosis of differentiating spermatogenic cells and disruption of blood-testis barrier function.

Astragalosides is the major active constituent of Radix Astragali. The present study was carried out to investigate the effect of crude astragalosides fraction (CAF) on rats liver fibrosis and its possible mechanisms. Hepatic fibrosis was induced by subcutaneous injection with 50% CCl(4) in Sprague-Dawley rats. The amount of CCl(4) administered was 1 mg kg(-1). The alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels in plasma and hydroxyproline (Hyp), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) contents in liver tissue were assayed by spectrophotometry. The hyaluronic acid (HA) and procollagen III (PC III) were assessed by radioimmunoassay. Tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) levels in culture supernatants of Kupffer cells (KCs) were determined with ELISA. Liver samples collected after 8 weeks of CCl(4) treatment were stained with hematoxylin-eosin (HE) and massion, and scored. Intragastric administration of CAF (10, 20 and 40 mg kg(-1)) significantly decreased indices of liver and spleen, the serum transaminase activities, HA and PC III levels, and Hyp and MDA contents in liver tissue in rats of hepatic fibrosis. Decreased SOD and GSH-px levels were reversed after administration of CAF. Histopathological scores showed CAF had inhibitory effect on the progression of hepatic fibrosis. In the in vitro experiments, CAF significantly reduced TNF-alpha and TGF-beta1 levels in culture supernatants of KCs. The results showed CAF significantly inhibited the progression of hepatic fibrosis induced by CCl(4), and the inhibitory effect of CAF on hepatic fibrosis might be associated with its ability to scavenge free radical and inhibit the production of TNF-alpha and TGF-beta1 from activated KCs.

To observe the expression of leptin (LP) and its influence on pulmonary fibrosis in experimental rats with silicosis and the correlation between the expressions of leptin and HIF-1α.A total of 120 male Sprague-Dawley rats were randomly divided into normal control group, silicosis model group, LP intervention groups (consisting of LP5, LP10 and LP20 groups according to the concentration of LP). The rats in the normal control group were intratracheally administered 1 mL normal sodium, and the ones in the other groups were intratracheally given 1 mL SiO₂(40 mg/mL) suspension. The rats in LP intervention groups were intraperitoneally injected with leptin 5, 10, 20 ng/kg.d respectively from the first day. Six rats in each group were sacrificed on the 7th, 14th, 21th and 28th day. The expression of LP on the 7th, 14th, 21th and 28th day and the hydroxyproline content on the 28th day in rat lung tissues were measured by ELISA, and the expressions of HIF-1α and LP proteins in the lung tissues of the silicosis model group were measured by Western blotting.There were significant differences in the expression of LP in the lung tissues at each time point (7th, 14th, 21th and 28th day) among the five groups (F=669.18, 948.67, 1 172.00, 521.55, P0.05). The LP content in the silicosis model group was significantly higher than that in the normal control group (P0.05) and the expression of LP in the LP intervention groups were significantly elevated at each time point (P0.05) as compared with the silicosis model group. Hydroxyproline content on 28th day was (0.89 ± 0.16), (3.14 ± 0.40), (3.78 ± 0.27), (4.35 ± 0.13), (4.87 ± 0.16) mg/g in normal control group, silicosis model group, LP5, LP10 and LP20 intervention groups, respectively. Compared with the normal control group, hydroxyproline content in the silicosis model group significantly increased (P0.05), and compared with the silicosis model group, hydroxyproline content in LP intervention groups were significantly raised (P0.05). The significant positive correlations were noted between the expressions of HIF-1α and LP (r=0.876, 0.796, 0.859 and 0.854 at each time, P0.05).The expression of LP goes up significantly in the lung tissues of rats with silicosis. Addition of exogenous leptin can increase the collagen deposition in the lung tissues; meanwhile leptin has a positive correlation with HIF-1α.

Objective To investigate the effect and the relevant potential mechanism of nonpeptide neurokinin 1 (NK1) receptor antagonist L-703,606 in the edema formation after burn injury.MethodL-703,606 treatment was performed in Sprague-Dawley (SD) rats at early stage after deep partial-thickness skin scalding. One hundred and fifty two adult male SD rats were used in the study and randomly divided into sham scald (SS, n=8), scald control (SC, n=48), and L-703,606 treatment (LT, n=48) groups. The rats in SC and LT groups were subjected to 20% total body surface area (TBSA) deep partial-thickness skin scalding. Modified Evans blue extravasation, tracing electron microscopy by lanthanum nitrate and mean water content assay were employed to observe and detect the changes of vascular permeability, ultrastructure and edema formation in adjacent tissue to the wounds and in the jejuna of rats at early stage (72 h) after scald.ResultsThe pathological increase of vascular permeability in the periwound tissue and jejunum of rats in LT group were significantly lower than that in SC group (P

To probe into the best acupoint-prescription for the simple acupoint catgut embedding therapy in treatment of bronchial asthma.Forty-eight Wistar rats were randomly divided into six groups: the simple acupoint catgut embedding at Feishu (BL 13), Danzhong (CV 17) and Shenshu (BL 23) group (group A), the simple acupoint catgut embedding at Shenshu (BL 23) group (group B), the simple acupoint catgut embedding at Feishu (BL 13) and Danzhong (CV 17) group (group C), Dexamethasone group (group D), model group (group E), and control group (group F), 8 rats in each group. The asthmatic models were established by Ovalbumin (OVA) except group F. Rats in group A, B and C were treated with catgut embedding at the corresponding acupoints from the first experimental day. In group E and D, Dexamethasone and sterile were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistantly. No interventions were added on group F. For the six groups, the symptoms of asthmatic attack were observed and the pathologic changes of lung tissue were examined.(1) The times of sneeze and rhinocnesmus scratching nose in group E were increased significantly compared with those in group F (both P0.01), but there was no significant difference between group D and F (both P0.05). As compared with that in group E, except the times of sneeze in group C, the times mentioned above were decreased significantly in all the treatment groups (all P0.01). There was no significant difference between group A and D (both P0.05), but the times mentioned above were increased significantly in group B and C as compared with that in group A (all P0.01). (2) The symptoms of asthmatic attack and the pathologic changes of airway tissue were all alleviated in group A, B and C, but a better amelioration was observed in group A, with no mucus epistom in the bronchial lumen and few infiltrations of inflammatory cells around the bronchi.The improvement of the simple acupoint catgut embedding at "Feishu" (BL 13), "Danzhong" (CV 17)and "Shenshu" (BL 23) on the airway inflammation in asthmatic rats is better than that of catgut embedding at "Feishu" (BL 13) and "Danzhong" (CV 17) or at "Shenshu" (BL 23) only.

Periodontal diseases, which are characterized by destruction of the connective tissues responsible for restraining the teeth within the jaw, are the main cause of tooth loss. Periodontal regeneration mediated by human periodontal ligament stem cells (hPDLSCs) may offer an alternative strategy for the treatment of periodontal disease. Dogs are a widely used large-animal model for the study of periodontal-disease progression, tissue regeneration, and dental implants, but little attention has been paid to the identification of the cells involved in this species. This study aimed to characterize stem cells isolated from canine periodontal ligament (cPDLSCs). The cPDLSCs, like hPDLSCs, showed clonogenic capability and expressed the mesenchymal stem cell markers STRO-1, CD146, and CD105, but not CD34. After induction of osteogenesis, cPDLSCs showed calcium accumulation in vitro. Moreover, cPDLSCs also showed both adipogenic and chondrogenic potential. Compared with cell-free controls, more cementum/periodontal ligament-like structures were observed in CB-17/SCID mice into which cPDLSCs had been transplanted. These results suggest that cPDLSCs are clonogenic, highly proliferative, and have multidifferentiation potential, and that they could be used as a new cellular therapeutic approach to facilitate successful and more predictable regeneration of periodontal tissue using a canine model of periodontal disease.

Tooth agenesis is a common craniofacial congenital malformation in humans, but little is known about the mechanisms of root resorption in this condition. The purpose of this study was to investigate the mechanisms of root resorption in primary molars without successors. An animal model without permanent tooth germs was established by surgery in beagles. The times of onset of primary molar root resorption, with and without successors, were compared. The distribution of immune cells, odontoclasts, and their activating factors were determined by histochemistry and immunohistochemistry. Root resorption of primary mandibular molars without successors began later than physiological resorption. In primary molars without permanent germs, odontoclasts and immune cells were present mainly in the apical pulp at the start of root resorption, whereas in control teeth receptor activator of nuclear factor-κB ligand (RANKL)-positive cells were found mainly in the region of the periodontal ligament. CD14(+) and CD3(+) cells were found in both the pulp and the periodontal ligament region. These results suggest that the dental pulp of primary molars, as well as immune cells, may play an important role in root resorption in primary molars without permanent tooth germs.

Hydroxysafflor yellow A (HSYA) is a main chemical component of the flower of Carthamus tinctorius. The present study investigated whether HSYA could attenuate brain injury induced by lymphostatic encephalopathy (LE). This was induced in adult male Wistar rats by cervical lymphatic blockade (CLB). Heart rate variability (HRV) was used as an indirect measurement of the regulatory function of the autonomic nervous system by recording the ECG signals from rats. It was shown that treatment with HSYA (5 mg/kg, i.p.) significantly alleviated the neurological deficits observed in rats with LE. Histological staining revealed that HSYA treatment attenuated LE-induced cell apoptosis in the rostral ventrolateral medullus (RVLM). Animals in the LE groups exhibited impaired regulatory roles of the autonomic nervous system in cardiovascular function, which was suppressed by pretreatment with HSYA. Additionally, HSYA administration significantly prevented the decrease of endothelial nitric oxide synthase (eNOS) mRNA and protein expression in the RVLM of rats with LE. These findings suggest that HSYA might provide neuroprotection against LE-induced brain injury and the associated functional alterations, which is likely regulated by the nitric oxide pathway.

To investigate the role of pulp in the root resorption of primary teeth without permanent tooth germs.The animal model without permanent tooth germs was established by surgery in Beagle dog. The root resorption was observed by taking periapical radiographs periodically. The samples of mandibular bone and pulp at different resorption stages were collected. The distribution of odontoclasts and the activating factor was analyzed by histological staining and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The role of pulp in the root resorption of primary teeth was tested by early pulpectomy.In the root resorption of primary molars without permanent teeth germs, a large number of odontoclasts were present on the pulpal surface of the root canal. Semi-quantification RT-PCR showed that the ratios of the expression of receptor activator of NF-κB ligand (RANKL) mRNA and β-actin in the pulp of permanent teeth and primary teeth without permanent teeth germ during different periods of root resorption are 0.1314, 0.1901, 0.2111 and 0.6058 (P0.05). The root resorption of primary teeth without permanent teeth germs in test groups was about 5 weeks later than that of control group.The pulp of primary tooth played an important role in the root resorption of primary tooth without permanent tooth germ.

To explore the underlying mechanism of the simple acupoint catgut emhedding on the treatment of the asthma in rats.Thirty two Wistar rats with asthma were estahlished hy ovalhumin (OVA), and randomly divided into four groups: the simple acupoint catgut emhedding group (A), Dexamethasone group (B), model group (C), control group (D), 8 cases in each group. In group A, emhed thread was carried out on the first experimental day, "Feishu (BL 13)" "Danzhong (CV 17)" "Shenshu (BL 23)" were selected. In the group B and the group C, 1.5 mg/kg Dexamethasone and 20 mg/kg sterile water were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistently. There was no treatment in the group D. For the four groups, asthmatic attack was ohserved, the cells in hronchoalveolar lavage fluid were counted and classified, the pathologic change of lung tissue was examined, the level of ICAM-1 and NF-kappaB in hronchioles was detected with immunohistochemistry.(1) In the group A, the symptom of asthmatic attack was alleviated and the infiltration of inflammatory cells around the hronchi were remarkahly decreased. In the group C, the total amount of cells in hronchoalveolar lavage fluid and other inflammatory cells were ohviously higher than those of group A. On the other hand, inflammatory effusion in airway and infiltration of inflammatory cells in airway wall in the group A were lower than those of group C. (2) The expression of ICAM-1 and NP-kappaB in lung tissue of group C was higher than those of group D (P0.01). (3) The expression of ICAM-1 and NF-kappaB in lung tissue of group B and group A was ohviously lower than those of group C (P0.05, P0.01).The simple acupoint catgut emhedding can inhahit the expression of ICAM-1 and NP-kappaB and alleviate airway inflammation in the asthmatic rats.

To investigate the effect of curcumin on apoptosis of pulmonary fibroblasts from rats and the molecular mechanisms.Twenty 6 week old male SD rats were randomly divided into 2 groups with 10 rats in each group: the normal control group (NLF) and the model group (MLF). The pulmonary fibrosis models were made by intratracheally instillation of bleomycin. The rats were sacrificed on the 28 day of experiment, and the pathologic changes of the lung tissues were investigated. Fibroblasts were derived from lung tissues and were cultured in vitro. Then the cells were treated with curcumin at different concentrations (5, 10, 20 and 40 micromol/L). TUNEL staining was used to detect apoptosis. The Caspase-3 and Caspase-8 expression of each group were determined by flow cytometry (FCM). The expression changes of apoptosis-regulating proteins of Caspase-9 in cells were examined by immunohistochemical staining.In the MLF group treated with 4 different concentrations of curcumin for 12 h, apoptotic morphological changes were observed in the cells. Apoptotic index (AI) was (30.17 +/- 0.53)%, (37.20 +/- 0.25)%, (45.12 +/- 0.31)%, (51.90 +/- 0.27)% respectively, which were significantly higher than that of the control group without curcumin (4.59 +/- 0.19)%. The expression of Caspase-3 was (53.1 +/- 0.7)%, (60.5 +/- 0.6)%, (69.1 +/- 0.9)%, (74.9 +/- 0.7)%, which were significantly higher than that of the control group (5.5 +/- 0.8)%. The expression of Caspase-8 were (37.2 +/- 0.5)%, (46.8 +/- 0.4)%, (56.5 +/- 0.3)%, (67.3 +/- 0.3)%, which were significantly higher than that of the control group (4.6 +/- 0.6)%. The expression of Caspase-9 were (39.38 +/- 0.88)%, (47.51 +/- 0.24)%, (57.75 +/- 0.63)%, (68.36 +/- 0.29)%, which were significantly higher than that of the control group (4.92 +/- 0.34)%. Curcumin did not induce apoptosis of NLF, the expressions of Caspase-3, Caspase-8, Caspase-9 being not statistically different from that of the control group without curcumin.Curcumin can induce apoptosis of pulmonary fibroblasts in rats with bleomycin-induced pulmonary fibrosis and the mechanism may be related to the activation of Caspase-3, Caspase-8, and Caspase-9.