Study on the Mechanism of BMSCs in Regulating NF-κB Signal Pathway by Targeting miR-449a to Improve the Inflammatory Response to Peripheral Nerve Injury.

OBJECTIVE: To evaluate the mechanism of Bone Marrow Mesenchymal Stem Cells (BMSCs) in regulating NF-κB signal pathway by targeting miR-449a. METHODS: Stem cells were transfected by over-expressing and inhibiting miR-449a to detect the levels and viability of miR-449a in stem cells after transfection. Stem cells and neurons were co-cultured in vitro to evaluate the in vitro mechanism of stem cells over-expressing miR-449a on neurons. RESULTS: After the addition of neurons, the neuronal activity of miR-449a over-expression group increased significantly, the expression of NF-κB signal pathway proteins (IκBα, p50, and p65) decreased, and the inflammatory cytokines (TNF-α and IL-1ß) decreased significantly (P<0.05). In vivo experiments in rats also showed that rats were unresponsive, did not chirp or elude after being stimulated. After stem cell therapy, the weight and response of rats gradually returned to normal levels. miR-449a expression significantly increased in the stem cell + miR-449a over-expression group, expression of NF-κB signal pathway proteins (IκBα, p50, and p65) decreased, inflammatory cytokines (TNF-α and IL-1ß) significantly decreased, and cell activity significantly increased (P<0.05). CONCLUSIONS: BMSCs can modulate NF-κB signaling pathway by targeting miR-449a, so as to reduce the inflammatory response to peripheral nerve injury and repair nerve injury.

A novel catalase mimicking nanocomposite of Mn(II)-poly-L-histidine-carboxylated multi walled carbon nanotubes and the application to hydrogen peroxide sensing.

In this work, a novel enzyme-mimicking nanocomposite of Mn(II)-poly-L-histidine (PLH) functionalized carboxylated multi walled carbon nanotubes (CMWCNTs) was designed and synthesized. Based on the catalase-like activity of the nanocomposite, a non-enzymatic hydrogen peroxide (H2O2) biosensor was then established and explored for H2O2 electrochemical detection. The nanocomposite was characterized by Fourier transform infrared spectra, Raman spectroscopy, and transmission electron microscopy. Due to the enlarged effective surface area and the efficient electrocatalytic activity of the Mn(II)-PLH redox-active units, the obtained Mn(II)-PLH-CMWCNT electrode showed excellent electrocatalytic performance toward H2O2 disproportionation. Under the selected optimum conditions, the prepared biosensor exhibited highly sensitive response toward H2O2, and the response current had a good linear relationship between the response currents and H2O2 concentrations in the range of 0.002-1.0 mM, a low detection limit of 0.5 µM and a sensitivity of 464.18 µA mM-1 cm-2. With the good stability, reproducibility and selectivity, the proposed biosensor was successfully applied to the determination of H2O2 in real-life samples, and showed satisfactory results. In summary, the Mn(II)-PLH-CMWCNT nanocomposite could be a promising enzyme-mimicking nanomaterial for the researches of electrocatalysis, biosensing and relevant fields.

Facile fabrication of a 3,4,9,10-perylene tetracarboxylic acid functionalized graphene-multiwalled carbon nanotube-gold nanoparticle nanocomposite for highly sensitive and selective electrochemical detection of dopamine.

A novel non-enzymatic electrochemical sensor for highly sensitive and selective detection of dopamine was developed based on a 3,4,9,10-perylene tetracarboxylic acid functionalized graphene-multiwalled carbon nanotube-gold nanoparticle nanocomposite modified glassy carbon electrode (PTCA-RGO-MWCNTs-Au NPs/GCE). The nanocomposite film was prepared by a facile, eco-friendly and controllable route and its morphology was characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopic (EDX) analysis, and X-ray diffraction (XRD) spectroscopy, respectively. Cyclic voltammetry and chronoamperometry were used for evaluating the electrochemical behaviors of the prepared sensor. The DA sensor exhibited excellent electrochemical performance toward DA with a sensitivity as high as 0.124 µA mM-1, a wide linear range of 1-100 µM and a low detection limit of 0.07 µM (S/N = 3). Moreover, it showed good selectivity toward DA without any obvious interference by AA and UA. Furthermore, the prepared DA sensor was applied to detect DA in real samples with satisfactory results.

Ginseng in white and red processed forms: Ginsenosides and cardiac side effects.

Ginseng (Panax ginseng Meyer) has long been consumed as a medicinal or functional food in East Asia. It is available as dried white ginseng (WG) and steamed red ginseng (RG), which might differ in ginsenoside profiles. We compared ginsenoside types of RG and WG using UPLC-MS/MS and evaluated how they biologically affected heart of healthy rats by recording electrocardiography, measuring biochemical indicators, analyzing cardiac tissue slides, and Ca2+ signaling pathways. About 25 and 29 ginsenosides were detected in WG and RG, respectively, and the total ginsenoside content of RG contained was nearly 1.8 times higher than that of WG. Among them, ginsenoside Rg4, ginsenoside Rg6, ginsenoside Rh4, ginsenoside Rk1, ginsenoside Rg5, and protopanaxadiol were detected only in RG, while 20(R)-ginsenoside Rg2 was detected only in WG. Male SD rats treated by intraperitoneal injection of WG or RG extracts were similar to the control in terms of electrocardiography and heart histology, indicating that both may not significantly affect the rats' myocardial function. However, WG and RG may induce mild cardiac injury resulting in increased cardiac collagen and creatine kinase levels. In addition, upregulated p-CaMKII and PPARδ and downregulated SERCA2a for WG and RG treatments were further associated with increased cardiac contractility. In general, RG had less effect on the heart of healthy rats than WG, which may be due to RG having a high proportion of low-polar ginsenosides.

Anti-senescence effects of Rhodiola crenulate extracts on LO2 cells and bioactive compounds.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola crenulata (Rc) is a traditional herb, used in Tibetan medicine, has shown promise efficacy in physical performance improvement, work capacity enhancement, fatigue elimination, and altitude sickness prevention. Also, Rc exhibited therapeutic effects on aging-related diseases. However, relevant researches on Rc and their bioactive components are quite few and needs further investigation. AIM OF THE STUDY: The objective of this study was to understand the relationship between phytochemical profiles and their activities of Rc extracts. MATERIALS AND METHODS: Rc extracts prepared by solvents with various hydrophilicity (i.e. aqueous ethanol (70%, v/v), water, and ethyl acetate), and their chemical compositions and specific compounds were analyzed by chemical analysis method and ultra-performance liquid chromatography quadruple time-of-flight mass spectrometry (UPLC-QTOF-MS). The regulate effects of Rc extracts on senescence and antioxidant activity were evaluated using the models of LO2 cells and Caenorhabditis elegans. RESULTS: The 70% ethanol extracts exhibited better regulating effects on senescence via the assays of senescence -associated ß-galactosidase (SAßG) staining and lifespan, which was consistent with the higher antioxidant activities observed based on the results of antioxidant assays. A total of 14 phytochemicals have been identified in 70% ethanol extracts, whereas the other two extracts contained much fewer compounds in varieties. Phytochemical profile of water extract was similar to the first half (polar compounds, running time: 0-6 min) of 70% ethanol extract profile, while those of ethyl acetate extract was consistent with its second half (more nonpolar compounds, running time: 6-12 min). CONCLUSIONS: The 14 phytochemicals in Rc might exhibit additive or synergistic effects on senescence regulating and antioxidant activities, providing theoretical basis for daily administration of Rc.

Infusion of haploidentical HSCs combined with allogenic MSCs for the treatment of ALL patients.

Although haploidentical stem cell transplantation (haplo-HSCT) offers almost all acute lymphoblastic leukaemia (ALL) patients an opportunity for immediate transplantation, it exhibits a higher incidence of graft failure and graft versus host disease (GVHD). Mesenchymal stem cells (MSCs) are characterised by their haematopoiesis-promoting and immunomodulatory capacity. Thus, we designed a combination of haplo-HSCT and MSCs for ALL patients. ALL patients (n = 110) were given haploidentical HSCs combined with allogenic MSCs, and ALL patients without MSC infusion (n = 56) were included as controls. The 100-day cumulative incidences of grade ≥2 acute GVHD (aGVHD) and grade ≥3 aGVHD were 40.00% and 9.09% compared to 42.32% (P = 0.79) and 22.79% (P = 0.03) in patients without MSC infusion, respectively. The 3-year cumulative incidences of chronic GVHD (cGVHD) and extensive cGVHD were 22.27% and 10.27% compared to 32.14% (P = 0.19) and 22.21% (P = 0.04) in patients without MSC infusion, respectively. No significant differences in the 3-year relapse incidence, nonrelapse mortality, leukaemia-free survival or overall survival in groups with and without MSC cotransplantation were observed. Multivariate analysis showed that MSC infusion contributed to a lower risk of developing extensive cGVHD. Our data suggested that haplo-HSCT combined with MSCs may provide an effective and safe treatment for ALL patients.

[Clinical Safety of NK Cell in the Prevention of Leukemia Relapse Post-transplantation and in Treatment of the Elderly Leukemia Patients].

OBJECTIVE: To observe the safety of donor NK cell infusions in the settings of hematopoietic stem cell transplantation and after consolidation chemotherapy in elderly patients with acute myeloid leukemia (AML). METHODS: Forty patients with AML were included, in which 21 patients aged over 60 years were at the stage of complete remission (CR) and 19 patients that received allogeneic hematopoietic stem cell transplantation (allo-HSCT). Mononucleated cells were isolated from peripheral blood from the donors (for allo-HSCT) or healthy immediate family members (elderly AML). The cells were seeded into the flasks pre-coated with NK cell specific activators, and expanded in media containing recombinant human IL-15 and IL-2 for 14 days. The cells were transfused intravenously after the identification of quality control. Trypan blue exclusion test was used for the determination of cell viability and counting. Flow cytometry analysis was performed to assess the surface antigenic profile. Seventy-eight infusions of the cell products were received by the elderly patients with AML after consolidation chemotherapy, 11 infusions were received by the patients during allo-HSCT and 32 infusions 3 moths after transplantation. The safety of cell therapy, body temperature, blood pressure and other indexes were observe during and 48 hours after cell transfusion. Meanwhile, the occurrence and severity of acute graft-versus-host disease (GVHD) were documented. RESULTS: Flow cytometry analysis showed that the proportion of NK cells (CD3-CD56+) in the mononucleated cells before culture was (14.10±4.22)% (n=121), and the proportion increased dramatically up to (87.29±8.75)% (n=121) after culture for 14 days, the number of NK cells increased to 753.47±140.13 times (n=121). The doses of the infused NK cells was (7.58±2.50)×107/kg per infusion. Moderate fever occurred in three cases after multiple infusions, and the temperature restored to normal on the same day after treatment. Fever was observed in one patient after every infusion of four times in total. The temperature reached to 38.5-39.0 ℃ and returned to normal within 1-2 hours after adequate antipyretic treatment, and then there was no discomfort. No GVHD was observed in the elderly AML patients, while 6 cases that received allo-HSCT developed moderate acute GVHD, among them grade I in 5 cases and grade II in 1 case. No other severe toxicities were observed. CONCLUSION: NK cell products with a high-purity could be obtained by ex vivo expansion with this protocol. The transfusion of these expanded cells is generally safe in the elderly patients with AML that have received chemotherapy or patients that received hematopoietic stem cell transplantation.

miR-539 Targeting SNAI2 Regulates MMP9 Signaling Pathway and Affects Blood-Brain Barrier Permeability in Cerebrovascular Occlusive Diseases: A Study Based on Head and Neck Ultrasound and CTA.

This study aimed to explore the expression level of miR-539 in the blood-brain barrier permeability induced by cerebrovascular occlusion and its mediated mechanism. Altogether, 48 patients with cerebral vascular occlusion lesions from March 2018 to June 2020 were collected. The expression level of miR-539 in the peripheral blood serum of the subjects was analyzed by qRT-PCR, and the participants were divided into two groups according to the results of head and neck ultrasound and CTA hemodynamics. The MCAO model of cerebral ischemia was established in rats, and the expression level of miR-539 was detected by qRT-PCR in brain tissues of different groups of rats. The effects of miR-539 on the permeability of blood-brain barrier were investigated by intraventricular injection of agomiR-539 and antagomir-539. The model of blood-brain barrier was established by culturing brain microvascular endothelial cells and pericytes in vitro, and the changes of miR-539 expression level and permeability after glucose and oxygen deprivation were detected. The expression level of SNAI2/MMP9 signaling pathway protein in cells was detected by Western blot. Compared with the healthy control group, the expression level of miR-539 in peripheral blood of patients with cerebrovascular occlusive disease decreased significantly, and the expression level of miR-539 in the MCAO rat model decreased and affected the permeability of blood-brain barrier. Glucose and oxygen deprivation treatment in brain microvascular endothelial cells can lead to downregulation of miR-539 expression and affect cell permeability. miR-539 in brain microvascular endothelial cells can target and bind to SNAI2 and participate in the regulation of endothelial cell permeability by affecting the SNAI2/MMP9 signaling pathway. The results of this study suggested that circulating miR-539 in peripheral blood may be a potential marker for predicting blood-brain barrier permeability after ischemic stroke. More detailed studies are needed to determine its diagnostic value.

CACNA1C rs1006737 SNP increases the risk of essential hypertension in both Chinese Han and ethnic Russian people of Northeast Asia.

ABSTRACT: Voltage-gated Ca2+ channels play a key role in the regulation of arterial tone and blood pressure. The aim of this study was to determine whether the association of calcium voltage-gated channel subunit alpha1 C (CACNA1C) rs1006737 with essential hypertension (EH) exists in both Chinese Han and ethnic Russian populations of Northeast Asia. We used a case-control study of 2 ethnic groups in the same latitude geographical area to investigate the association between the susceptibility of EH and rs1006737 polymorphism. A total of 1512 EH patients and 1690 controls in Chinese Han people (Heilongjiang Provence, China), 250 EH patients, and 250 controls in ethnic Russian people (Chita, Russia), participated in this study. All participants were genotyped using the TaqMan SNP genotyping assay (Agena Company). Baseline characteristics and the minor allele frequencies of rs1006737 vary substantially among common Chinese Han and ethnic Russian people. Allele A was found to be a risk factor for EH in Chinese Han [(odds ratio) OR 1.705, (confidence interval) 95% CI: 1.332-2.182, P < .001] and ethnic Russian (OR 1.437; 95% CI: 1.110-1.860, P = .006). The GA genotype was significantly associated with an increased risk of hypertension (OR 1.538, 95% CI: 1.188-1.991, P = .001) for Chinese Han people, and the AA genotype (OR 2.412, 95% CI: 1.348-4.318, P = .003) for ethnic Russian people. The results of this study indicate that the A allele of the variant rs1006737 in the CACNA1C gene may be a useful genetic marker for EH risk prediction in Chinese Han and ethnic Russian populations.

Preventive effect of ethanol extract from Chinese sumac fruits against tetrachloromethane-induced liver fibrosis in mice.

Chinese sumac (Rhus chinensis Mill.) fruits are traditionally used as a condiment and herb. The aim of this study is to evaluate the preventive effect of the ethanolic extract from Chinese sumac fruits against CCl4-induced liver fibrosis in mice and to delineate the underlying mechanisms. The results showed that the ethanolic extract substantially decreased AST, ALT, MDA, and hydroxyprolin (HYP), restored GSH, SOD, and CAT, and prevented liver fibrosis induced by CCl4. Further, investigation revealed that the prevention against liver fibrosis may be related to heightened inflammation due to suppression of the release of IL-1ß, IL-6, and TNF-α, and the downregulation of COX-2, iNOS, p-NF-κB, and p-P38; the prevention of hepatocyte apoptosis was due to the upregulation of Bcl-2 and downregulation of Bax; the reduction of HSCs activation was due to the downregulation of TGF-ß1 and upregulation of PPAR-γ; and the decrease in the extracellular matrix (ECM) deposition was due to the regulation of MMP9 and TIMP2. These findings proved that Chinese sumac fruits could effectively prevent CCl4-induced liver fibrosis in mice and may be used to develop functional foods and/or nutraceuticals to prevent or alleviate liver fibrosis.

Phenolic profiles, antioxidant activities and cytoprotective effects of different phenolic fractions from oil palm (Elaeis guineensis Jacq.) fruits treated by ultra-high pressure.

The present work investigated the phenolic profiles, antioxidant activities, and cytoprotective effects of the free, esterified, and insoluble-bound phenolic fractions from oil palm fruits treated under ultra-high pressure (UHP). Results showed that UHP treatment significantly increased the total phenolic and flavonoid contents of all three phenolic fractions (p < 0.05). A total of 11 and 12 phenolic compounds were detected and quantified in non-treated and UHP-treated fruits, with caffeic acid having the highest concentration in insoluble-bound phenolic fractions with 8.68 and 11.27 mg/g of dry extract, respectively. The antioxidant activities, intracellular reactive oxygen species inhibition, and cytoprotective effects of all three phenolic fractions were dramatically enhanced after UHP pretreatment (p < 0.05). Therefore, UHP-treated oil palm fruits with increased bioactivities could be used in functional food or the nutraceutical industry to enhance their applications and economic value.

Effects of Different Oligochitosans on Isoflavone Metabolites, Antioxidant Activity, and Isoflavone Biosynthetic Genes in Soybean ( Glycine max) Seeds during Germination.

Five oligochitosans with increasing degrees of polymerization (DPs), i.e., from chitotriose to chitoheptaose, were examined to clarify the structure-bioactivity relationship between the DPs of oligochitosans and their effects on the isoflavone metabolites, total phenolic and flavonoid contents (TPC and TFC, respectively), and antioxidant activity of soybean ( Glycine max) seeds during germination. Oligochitosans of different DPs exhibited varying influences on the TPC, TFC, and antioxidant activities of soybean seeds. Chitohexaose exerted a strong effect and significantly increased the aforementioned parameters in soybean seeds 72 h after germination. Genistin, malonylgenistin, and genistein were the main isoflavones found, and the genistin and genistein contents were significantly enhanced by 67.32% and 131.38%, respectively, after chitohexaose treatment. Several critical genes involved in the isoflavone biosynthesis (i.e., PAL, CHS, CHI, IFS) of soybeans treated with and without chitohexaose were analyzed, and results suggested that chitohexaose application could dramatically stimulate the transcription of these genes.

Correlations of Post-stroke Depression with Inflammatory Response Factors.

BACKGROUND: We aimed to investigate the correlations of the inflammatory response factors, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), high-sensitivity C-reactive protein (CRP), with patients with post-stroke depression (PSD), so as to provide a basis for the treatment and prevention of PSD for patients. METHODS: The clinical laboratory data of 60 patients with PSD in The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, China from July 2016 to July 2017 and those of another 60 stroke patients without PSD admitted in the same period were analyzed retrospectively. The expression levels of inflammatory response factors in the two groups of patients and in PSD patients with different levels of depression were compared and analyzed via statistical methods. Multiple Logistic regression analysis was used to determine whether inflammatory response factors were independent risk factors for PSD patients. RESULTS: The expression levels of IL-6, TNF-α and CRP in patients with PSD were significantly increased compared with those in patients without PSD, and the differences were statistically significant (t=6.429, t=6.355, t=5.792, P<0.001). The levels of IL-6, TNF-α and CRP had statistically significant differences between any two groups of mild, moderate and severe PSD patients (P<0.05). Results of multiple Logistic regression analysis revealed that the odds ratio (OR) values of inflammatory factors (IL-6, TNF-α and CRP) were 1.160, 1.099 and 1.248, respectively, and the corresponding p values were 0.020, 0.039 and 0.007 in patients of observation group, indicating the above three inflammatory response factors were independent risk factors for PSD. CONCLUSION: The clinic control on the expression levels of inflammatory response factors (IL-6, TNF-α and CRP) are extremely important for the treatment and prevention of PSD.

Facile Synthesis of ß-Lactoglobulin-Functionalized Reduced Graphene Oxide and Trimetallic PtAuPd Nanocomposite for Electrochemical Sensing.

The use of graphene has leapt forward the materials field and the functional modification of graphene has not stopped. In this work, ß-lactoglobulin (BLG) was used to functionalize reduced graphene oxide (RGO) based on its amphiphilic properties. Also, trimetallic PtAuPd nanoparticles were reduced to the surface of BLG-functionalized RGO and formed BLG-PtAuPd-RGO nanocomposite using facile synthesis. Transmission electron microscopy, energy-dispersive X-ray spectroscopy and Fourier transform infrared spectra were used to characterize the nanocomposite. Electrocatalytic analysis was evaluated through cyclic voltammetry and chronoamperometry methods. We developed a glucose sensor by fabricating GOD-BLG-PtAuPd-RGO/glassy carbon (GC) electrode. It presented a remarkable sensitivity of 63.29 µA mM-1 cm-2 (4.43 µA mM-1), a wider linear range from 0.005 to 9 mM and a lower detection limit of 0.13 µM (S/N = 3). Additionally, the glucose sensor exhibited excellent testing capability in human serum samples.

Copper (II)-ploy-L-histidine functionalized multi walled carbon nanotubes as efficient mimetic enzyme for sensitive electrochemical detection of salvianic acid A.

In this work, carboxylated multi walled carbon nanotubes (CMWCNTs) were firstly prepared and functionalized with poly-L-histidine (PLH), which were then chelated with copper (II) ions to from the nanocomposites of Cu(II)-PLH-CMWCNTs. The nanocomposites could be exploited as an efficient mimic enzyme for sensitive electrochemical detection of salvianic acid A (SAA). Cu(II)-PLH-CMWCNTs owned good charge transfer property and excellent synergetic catalytic effect between the overoxidized imidazole groups and the copper redox-active units. Therefore, highly sensitive electrochemical response to SAA was achieved under optimum experimental conditions. A good linear relationship between differential pulse voltammetry (DPV) peak current and the SAA concentration was established in the range of 0.4-1000 µM. A low detection limit of 0.037 µM and a sensitivity of 0.27 µA µM-1 cm-2 were achieved. The developed biosensor also had advantages of good repeatability, stability and high selectivity, thus, it was successfully applied to the determination of SAA in real samples with satisfactory results, which may have great potential for further exploitation of electroanalysis applications.