Ginsenoside Rg1 (Rg1) has been shown to treat a variety of human diseases, including Alzheimer’s disease (AD). However, its mechanism in AD needs further investigation. Microglial cells (BV2) were treated with Aβ1-42 to induce AD cell models. Cell viability and apoptosis were tested by cell counting kit 8 assay and flow cytometry. The protein levels of GATA-binding protein 4 (GATA4), phosphodiesterase 4A (PDE4A), autophagy-related markers, M1/M2 polarization-related markers and PI3K/AKT-related markers were detected by western blot. Inflammation factors were detected by ELISA. Jaspar and dual-luciferase reporter assay were used to evaluate the interaction between GATA4 and PDE4A. Our results showed that Rg1 promoted viability and autophagy, while suppressed apoptosis and inflammation in Aβ1-42-induced BV2 cells. Rg1 reduced GATA4 protein expression, and GATA4 upregulation reversed the regulation of Rg1 on Aβ1-42-induced BV2 cell injury. GATA4 interacted with PDE4A, and GATA4 facilitated Aβ1-42-induced BV2 cell injury by increasing PDE4A expression. Besides, GATA4 knockdown reduced PDE4A protein expression and inactivated PI3K/AKT axis, while these effects were abolished by PDE4A overexpression. In conclusion, our data suggested that Ginsenoside Rg1 inhibited microglial cell apoptosis and inflammation to attenuate AD progression by regulating the GATA4/PDE4A/PI3K/AKT axis.

<p>Supplementary Table1.Characteristic of antibodies used in the flow cytometry evaluation.Supplementary Table2.Primer sequence and product size for PCR amplification.Supplementary Table3.Clinical and baseline clinical characteristics of OC patients with diabetes</p>

<p>Supplementary Fig.1.Increased MDSCs levels in peripheral blood and tumor from OC patients.Supplementary Fig.2.Characterization of phenotype and immunosuppressive activity of MDSCs in OC patients.Supplementary Fig.3.Phenotypic analysis of MDSCs following metformin treatment.</p>

Editor's Note: this Article has been retracted; the Retraction Note is available at https://doi.org/10.1038/s41598-021-83145-9.

RNA N6-methyladenosine (m6A) level is closely associated with neurodevelopment and central nervous system dysfunctions including spinal cord injury (SCI). M6A level can be dynamically regulated by m6A methyltransferases and demethylases. In this text, the roles of m6A demethylase FTO alpha-ketoglutarate dependent dioxygenase (FTO) in SCI development along with its m6A-dependent regulatory mechanisms were investigated in hypoxia-induced PC12 cell injury model. The results showed that FTO was low expressed in spinal cord tissues of rats after contusive SCI and hypoxia-treated PC12 cells. FTO knockdown alleviated hypoxia-induced PC12 cell injury. FTO loss increased GADD45B expression and m6A level in PC12 cells. GADD45B knockdown weakened the protective effects of FTO depletion on hypoxia-treated PC12 cells. FTO regulated GADD45B expression in an IGF2BP2-dependent manner. In conclusion, FTO knockdown mitigated the injury of hypoxia-induced PC12 cells by up-regulating GADD45B in an IGF2BP2-dependent manner.

Ginsenoside Rb1 (GRb1), a major ingredient of ginseng, has been found to be a potential protective agent in spinal cord injury (SCI) and in activated microglia-induced neuronal injury. This study discovered that GRb1 could facilitate miR-130b-5p expression in SCI rats and Toll-like receptor 4 (TLR4; a crucial player in inflammation) was a potential target of miR-130b-5p. Hence, we further investigated whether GRb1 could relieve SCI by reducing microglia-mediated inflammatory responses and neuronal injury via miR-130b-5p/TLR4 pathways. The results showed that GRb1 alleviated SCI through inhibiting neuronal apoptosis and proinflammatory factor expression via increasing miR-130b-5p.GRb1 weakened the damage of activated microglia to neurons through upregulating miR-130b-5p. miR-130b-5p attenuated activated microglia-induced neuron injury via targeting TLR4. GRb1 inactivated TLR4/nuclear factor-kappa B (NF-kappa B) activation and inhibited proinflammatory cytokine secretion by increasing miR-130b-5p in activated microglia. As a conclusion, GRb1 alleviated SCI through reducing activated microglia-induced neuronal injury via miR-130b-5p/TLR4/NF-kappa B axis, providing a deep insight into the molecular basis of GRb1 in the treatment of SCI.

脊髓损伤(spinal cord injury,SCI)是一种严重的神经系统疾病,其特点为损伤部位神经元功能的永久丧失及胶质瘢痕形成,导致相应支配区域的感觉和运动功能障碍.SCI可能会引起一系列严重后果,例如感觉障碍和肢体瘫痪;另外,由于失去了神经的营养作用,骨质疏松和肌肉萎缩也是其常见的并发症[1].SCI不仅给患者及家庭带来沉重的生理和心理负担,而且给社会造成巨大的经济损失,故而其预防、治疗和康复已成为医学领域的重要问题.SCI的发病机制复杂,涉及原发性和继发性损伤两个阶段.在骨折、压迫等原发性损伤发生后,炎症反应、组织缺氧、神经元坏死和凋亡、局部微环境抑制等一系列继发性损伤进一步加重了脊髓损伤,导致严重的感觉和运动功能丧失.临床上,即使患者能得到及时的救治,但仍有相当一部分SCI患者终生忍受着瘫痪的折磨[2].因此,寻找能够有效治疗该疾病的新的分子靶标至关重要.

椎弓根螺钉(pedicle screw,PS)因能够固定脊柱三柱达到稳定的三维固定效果,自其发明以来一直广泛应用于各种脊柱外科手术.然而螺钉松动、断裂、拔出等各种并发症往往导致固定失败,尤其对于骨质疏松的患者,螺钉松动是最为常见的并发症[1、2].因此,对于骨量下降的患者,如何提高PS的把持力及稳定性是一个亟待解决的问题.临床医师往往通过延长脊柱固定节段或结合骨水泥强化钉道等方法提高骨质疏松患者脊柱内固定的稳定性[3、4].然而这些方法仍存在诸多不足,如:骨水泥使用过程中存在高温释放引起脊髓或神经根损伤、骨水泥疲劳断裂、骨水泥外溢和栓塞等问题,延长脊柱固定节段增加了手术时间、出血量及其他并发症.为了克服以上缺点,基于传统PS钉道,Santoni等[5]首次提出了椎弓根皮质骨轨迹(cortical bone trajectory,CBT)螺钉固定技术.据报道,术中CBT螺钉置入扭矩是传统PS的1.71倍[6].另有研究也表明,CBT螺钉固定较传统PS固定具有更佳的抗轴向拔出力[7、8].此外,和传统PS固定相比,CBT螺钉的进钉点更偏内侧且钉道和皮质骨接触面积更大,具有更好的界面强度和微创等优点[5].但该技术出现后国内外相关文献报道较少,笔者就其临床应用及研究进展进行综述,以期为骨质疏松等患者群体提供最佳的治疗方案.

椎间盘退变(intervertebral disc degeneration,IDD)是引起腰腿痛最常见的慢性脊柱退行性病变,目前尚无较好的非手术治疗方法[1].IDD的发生发展是一个多因素共同作用的过程,如遗传、衰老、外伤、过度负重、吸烟等,具体机制目前仍未完全阐明.有学者提出,细胞因子/蛋白质注射、细胞移植和基因转入是治疗IDD的三个潜在策略[2-5],但目前的研究尚存在诸多问题.

Microenvironmental factors such as oxygen concentration mediate key effects on the biology of mesenchymal stromal cells (MSCs). Herein, we performed an in-depth characterization of the metabolic behavior of MSCs derived from the placenta, umbilical cord, and adipose tissue (termed hPMSCs, UC-MSCs, and AD-MSCs, respectively) at physiological (hypoxic; 5% oxygen [O2]) and standardized (normoxic; 21% O2) O2 concentrations using chemical isotope labeling liquid chromatography-mass spectrometry. 12C- and 13C-isotope dansylation (Dns) labeling was used to analyze the amine/phenol submetabolome, and 2574 peak pairs or metabolites were detected and quantified, from which 52 metabolites were positively identified using a library of 275 Dns-metabolite standards; 2189 metabolites were putatively identified. Next, we identified six metabolites using the Dns library, as well as 14 hypoxic biomarkers from the human metabolome database out of 96 altered metabolites. Ultimately, metabolic pathway analyses were performed to evaluate the associated pathways. Based on pathways identified using the Kyoto Encyclopedia of Genes and Genomes, we identified significant changes in the metabolic profiles of MSCs in response to different O2 concentrations. These results collectively suggest that O2 concentration has the strongest influence on hPMSCs metabolic characteristics, and that 5% O2 promotes arginine and proline metabolism in hPMSCs and UC-MSCs but decreases gluconeogenesis (alanine-glucose) rates in hPMSCs and AD-MSCs. These changes indicate that MSCs derived from different sources exhibit distinct metabolic profiles.

Protective effect of Qishen Yiqi dropping pills on the myocardium of rats with chronic heart failure (CHF) was investigated. Sixty rats were divided into the sham operation (n=20), the model (n=20) and the Qishen Yiqi dropping pill treatment group (n=20) using the random table method. The treatment group received administration of Qishen Yiqi dropping pills. The model and the sham operation group were given the same amount of normal saline. Within 24 h after the last administration, the rats were sacrificed. The myocardia were used for reverse transcription-polymerase chain reaction, western blot analysis and histological examination. In the sham operation group, cardiomyocytes were stained evenly and arranged neatly and densely with clear structures. In the model group, the cell morphology was fuzzy, the myocytes were hypertrophied, the nuclear pyknosis was fragmented, the arrangement was disordered, the intercellular space was narrowed, and the cytoplasm was missing. The apoptosis rates of cardiomyocytes in the model and Qishen Yiqi dropping pill treatment group were significantly higher than that in the sham operation group (P<0.05). The myocardial infarction areas in the model group and the Qishen Yiqi dropping pill treatment group were larger than that in the sham operation group (P<0.05). The expression levels of transforming growth factor-β1, mothers against decapentaplegic homolog 2 (Smad2), Smad3, and caspase-3 messenger ribonucleic acids and proteins in the model group and the Qishen Yiqi dropping pill treatment group were higher than those in the sham operation group (P<0.05). Qishen Yiqi dropping pills have an obvious myocardial protective effect on CHF rats, which may enhance the degree of myocardial fibrosis by inhibiting the TGF-β1/Smads pathway and improve cardiomyocyte apoptosis by suppressing the caspase-3 signaling pathway, thus protecting the myocardium.

The placenta resides in a physiologically low oxygen microenvironment of the body. Hypoxia induces a wide range of stem cell cellular activities. Here, we report a workflow for exploring the role of physiological (hypoxic, 5% oxygen) and original cell culture (normoxic, 21% oxygen) oxygen concentrations in regulating the metabolic status of human placenta-derived mesenchymal stem cells (hPMSCs). The general biological characteristics of hPMSCs were assessed via a variety of approaches such as cell counts, flow cytometry and differentiation study. A sensitive 13C/12C-dansyl labeling liquid chromatography-mass spectrometry (LC-MS) method targeting the amine/phenol submetabolome was used for metabolic profiling of the cell and corresponding culture supernatant. Multivariate and univariate statistical analyses were used to analyze the metabolomics data. hPMSCs cultured in hypoxia display smaller size, higher proliferation, greater differentiation ability and no difference in immunophenotype. Overall, 2987 and 2860 peak pairs or metabolites were detected and quantified in hPMSCs and culture supernatant, respectively. Approximately 86.0% of cellular metabolites and 84.3% of culture supernatant peak pairs were identified using a dansyl standard library or matched to metabolite structures using accurate mass search against human metabolome libraries. The orthogonal partial least-squares discriminant analysis (OPLS-DA) showed a clear separation between the hypoxic group and the normoxic group. Ten metabolites from cells and six metabolites from culture supernatant were identified as potential biomarkers of hypoxia. This study demonstrated that chemical isotope labeling LC-MS can be used to reveal the role of oxygen in the regulation of hPMSC metabolism, whereby physiological oxygen concentrations may promote arginine and proline metabolism, pantothenate and coenzyme A (CoA) biosynthesis, and alanine, aspartate and glutamate metabolism.

Objective To assess the value of serum carcinoembryonic antigen (CEA)and carbohydrate antigen 19-9 (CA19-9) levels in the diagnosis of gastric cancer in elderly patients.Methods This prospective study included 104 elderly patients with gastric cancer at our hospital from March 2015 to December 2016 to serve as an experimental group.Moreover,104 elderly patients with benign gastric lesions were enrolled to serve as a control group.Serum CEA and CA19-9 levels were compared between the two groups.The diagnostic specificity,sensitivity,and validity for gastric cancer were calculated by using CEA or CA19-9 alone or the two in combination.Results Serum CEA and CA19-9 levels in the experimental group were significantly higher than in the control group(t =3.001 and 5.110,P =0.039 and 0.016,respectively);The specificity,sensitivity,and validity of CEA and CA19-9 in combination were significantly higher than when each of the measures was used alone(x2 =2.101 and 2.109,P =0.031 and 0.019,respectively).Serum CEA levels showed good predictive power for both lymph node metastasis of gastric cancer(Z =5.109,P =0.002) and liver metastasis of gastric cancer(Z =3.910,P =0.026);Serum CA19-9 levels could predict lymph node metastasis of gastric cancer (Z =4.189,P =0.003) Conclusions Serum CEA and CA19-9 in combination have high sensitivity and validity for gastric cancer detection;Elevated CEA can predict gastric cancer metastasis and elevated CA19-9 may predict lymph node metastasis.

Spinal cord injury (SCI) usually results in neurological damage. DGCR5 is closely related to neurological disorders, and this study aims to explore its role in neuronal apoptosis in acute SCI. The ASCI model was established in rats, and the Basso, Beattie, and Bresnahan (BBB) scoring was used to assess the neurological function. The expression of RNA and protein was quantified by quantitative real-time PCR (qRT-PCR) and western blotting, respectively. The oxygenglucose deprivation (OGD) was performed upon neurons and apoptosis was evaluated by flow cytometry. The interaction and binding between DGCR5 and PRDM5 was detected with RNA pull-down and RIP assay, respectively. DGCR5 was down-regulated in ASCI model rat and in neurons treated with hypoxia. Over-expression of DGCR5 inhibited neuronal apoptosis. Interaction between DGCR5 negatively regulated PRDM5 protein expression by binding and interacting with it. DGCR5 inhibited neuronal apoptosis through PRDM5. Over-expressed DGCR5 ameliorated ASCI in rat. DGCR5 suppresses neuronal apoptosis through directly binding and negatively regulating PRDM5, and thereby ameliorating ASCI.

Epilepsy is a common neurological disorder in the central nervous system. Inflammation disrupts the blood-brain barrier (BBB), which is responsible for maintaining brain homeostasis. This study was aimed to investigate the functional role of microRNA (miR)-132 in hippocampal HT-22 cells under lipopolysaccharide (LPS) stimulation. In vitro cell inflammatory model was constructed by LPS stimulation. Inflammatory cell injury was evaluated according to the alterations of cell viability, apoptosis, and expression of inflammatory cytokines. Then, miR-132 level after LPS treatment was assessed. Subsequently, miR-132 was abnormally expressed after cell transfection, and the effects of miR-132 on LPS-induced cell inflammatory injury as well as phosphorylated levels of key kinases in the NF-κB and MAPK kinase (MEK)/ERK pathways were determined. The target gene of miR-132 was virtually screened and verified, and whether miR-132 affected HT-22 cells under LPS stimulation through regulating the target gene was verified. The results showed that the level of miR-132 was down-regulated by LPS in HT-22 cells, and the LPS-induced inflammatory injury could be reduced by miR-132 overexpression. Then, the phosphorylated levels of kinases in the NF-κB and MEK/ERK pathways were decreased by miR-132 overexpression. Tumor necrosis factor receptor-associated factor 6 (TRAF6) was predicted and verified to be a target of miR-132. Moreover, the alterations induced by miR-132 overexpression in the LPS-treated HT-22 cells were abrogated by TRAF6 overexpression. Therefore, we drew the conclusion that LPS down-regulated miR-132 and miR-132 attenuated LPS-induced inflammatory cell injury by targeting TRAF6, along with the inhibition of the NF-κB and MEK/ERK pathways.

Melatonin has been shown to inhibit myocardial infarction-induced apoptosis, its function in heart failure with preserved ejection fraction (HFpEF) has not been investigated. This study aimed to investigate whether melatonin attenuates obesity-related HFpEF. Male mice were fed a high-fat diet (HFD) from weaning to 6 months of age to induce HFpEF. The mice were orally administered melatonin (50 mg/kg) by 3 weeks. Diastolic function was significantly improved by melatonin supplementation in mice fed an HFD. Melatonin attenuated obesity induced myocardial oxidative stress and apoptosis and promoted the secretion of C1q/tumour necrosis factor related protein 3 (CTRP3) by adipose tissue. And depletion of circulating CTRP3 largely abolished melatonin-mediated cardio-protection. Melatonin-mediated secretion of adipocyte-derived CTRP3 activated NF-E2-related factor 2 (Nrf2), which were largely abrogated by knocking down CTRP3 in adipocytes or Nrf2 in cardiomyocytes. Nrf2 activation was mediated by miR-200a, and a miR-200a antagomir offset the effects of melatonin-conditioned medium on Nrf2 expression. Our results indicate that melatonin can be used to treat and prevent obesity related HFpEF.

Introduction Green fluorescent protein (GFP) is widely used as a reporter gene in regenerative medicine research to label and track stem cells. Here, we examined whether expressing GFP gene may impact the metabolism of human placental mesenchymal stem cells (hPMSCs). Methods The GFP gene was transduced into hPMSCs using lentiviral-based infection to establish GFP+hPMSCs. A sensitive 13C/12C-dansyl labeling LC-MS method targeting the amine/phenol submetabolome was used for in-depth cell metabolome profiling. Results A total of 1151 peak pairs or metabolites were detected from 12 LC-MS runs. Principal component analysis and partial least squares discriminant analysis showed poor separation, and the volcano plots demonstrated that most of the metabolites were not significantly changed when hPMSCs were tagged with GFP. Overall, 739 metabolites were positively or putatively identified. Only 11 metabolites showed significant changes. Metabolic pathway analyses indicated that three of the identified metabolites were involved in nine pathways. However, these metabolites are unlikely to have a large impact on the metabolic pathways due to their nonessential roles and limited hits in pathway analysis. Conclusion This study indicated that the expression of ectopic GFP reporter gene did not significantly alter the metabolomics pathways covered by the amine/phenol submetabolome.

BACKGROUND Nutrition support is crucial for patients with gastrointestinal (GI) cancer after the operation. However, the controversy over the application of parenteral nutrition (PN) and early enteral immunonutrition (EEIN) has no determinate conclusion. MATERIALS AND METHODS We compared the effects of PN and EEIN on the postoperative nutrition condition, immune status, inflammation level, long-term survival, and quality of life of the patients with GI cancer. Seventy-eight patients were randomly divided into the PN group (n = 44) or EEIN group (n = 34). After an 8-day nutrition treatment, clinical and immunological parameters were evaluated. RESULTS The EEIN group had a significantly shorter hospital stay and higher body mass index level on postoperative day 30 than those in the PN group (P < .05). However, total hospital cost and incidences of short-term postoperative complications had no significant difference (P > .05). The percentages of CD4+ , natural killer, and natural killer T lymphocyte cells and the ratio of CD4+ /CD8+ in peripheral blood were significantly increased. Compared with the PN group, the EEIN group had a higher expression of activated cell surface markers such as CD27 and CD28. In addition, the secretion of interleukin (IL)-2 and interferon-γ was significantly higher, and the secretion of tumor necrosis factor-α and IL-10 was lower. Complication-free survival in the EEIN group were longer than those in the PN group (P = .04). CONCLUSION EEIN is superior to PN in improving nutrition status, enhancing immune function, and elevating quality of life.

A computer-aided 3-dimensional (3D) visualization operation simulation system based on computer-aided design (CAD) Unigraphics NX and Mimics software was established to provide orthopedic surgeons with an actual and reliable system in treating of distal femoral fracture.According to the preoperative CT data, 3D reconstruction of the distal femoral fracture could be achieved by the Mimics software. Then, the CAD Unigraphics NX software was used to measure the model function of all the related surgical instruments, including less invasive stabilization system (LISS) and retrograde intramedullary nail fixation.The function of CAD Unigraphics NX and Mimics software was successful in assisting in the treatment of distal femoral fracture with LISS and retrograde intramedullary nail fixation. The operation procedure was actual, visualized, and lifelike. Moreover, the operation effect could be estimated before surgery.The virtual surgery system may improve the reliability and safety of the operative care of distal femoral fracture.

Background. Liver fibrosis is a chronic progressive liver disease, but no established effective treatment exists except for liver transplantation. The present study was designed to investigate the effect of human placenta mesenchymal stem cells (hPMSCs) expressing green fluorescent protein (GFP) on carbon tetrachloride- (CCl4-) induced liver fibrosis in rats. Methods. Liver fibrosis was induced by subcutaneous injection with CCl4; hPMSCs were directly transplanted into rats through the caudal vein. The therapeutic efficacy of hPMSCs on liver fibrosis was measured by liver function tests, liver elastography, histopathology, Masson's trichrome and Sirius red staining, and immunohistochemical studies. The expression levels of fibrotic markers, transforming growth factor β1 (TGF-β1) and α-smooth muscle actin (α-SMA), were assessed using real-time polymerase chain reaction. Results. We demonstrated that liver fibrosis was significantly dampened in the hPMSC transplantation group according to the Laennec fibrosis scoring system and histological data. The Sirius red-stained collagen area and the elastography score were significantly reduced in the hPMSC-treated group. Meanwhile, hPMSC administration significantly decreased TGF-β1 and α-SMA expression and enhanced liver functions in CCl4-induced fibrotic rats. Conclusion. This study indicates that transplantation of hPMSCs could repair liver fibrosis induced by CCl4 in rats, which may serve as a valuable therapeutic approach to treat liver diseases.