Objective:To discuss the effects of bone marrow-derived mesenchymal stem cells(BMSCs)transplantation on mitophagy in the vascular dementia(VaD)rats through reactive oxygen species(ROS)/nuclear factor erythroid 2-related factor 2(Nrf2)signaling,and to clarify its mechanism.Methods:Forty-five male adult SD rats were randomly divided into sham operation group,model group,unloaded group,BMSCs group,and MSCs+ML385(Nrf2 inhibitor)group(combination group),and there were 9 rats in each group.After intraperitoneal anesthesia,the VaD models were established in all groups except sham operation group.Morris water maze test was used to detect the learning and memory abilities of the rats in various groups;HE staining was used to observe the histopathological morphology of brain tissue of the rats in various groups;Nissl staining was used to observe the changes of Nissl bodies in hippocampus region of brain tissue of the rats in various groups;transmission electron microscope was used to observe the ultrastructure of hippocampus region of the rats in various groups;fluorescence probe method was used to detect the ROS levels in hippocampus neurons in various groups;Western blotting method was used to detect the expression levels of Nrf2,heme oxygenase-1(HO-1),PTEN-induced putative kinase 1(PINK1),parkin RBR E3 ubiquitin protein ligase(Parkin),Beclin-1,ubiquitin-binding protein p62(P62),and microtubule-associated protein 1A/1B-light chain 3(LC3-Ⅱ/LC3-Ⅰ)ratio in brain tissue of the rats in various groups.Results:The Morris water maze results showed that compared with sham operation group,the escape latency of the rats in model group was significantly increased(P<0.01),while the number of crossing time and residence time were significantly decreased(P<0.01).Compared with model group,the escape latency of the rats in BMSCs group was significantly decreased(P<0.01),while the number of crossing time and residence time were significantly increased(P<0.01).Compared with BMSCs group,the escape latency of the rats in combination group was significantly increased(P<0.01),while the number of crossing time and residence time were significantly decreased(P<0.01).The HE staining results showed that hippocampus neurons of the rats in sham operation group were normal in quantity and morphology,with uniform staining and clear structure.Compared with sham operation group,the hippocampus tissue of the rats in model group showed sparse arrangement,disordered structure,reduced neuronal quantity,varied morphology,uneven staining,nuclear pyknosis,and partial neuronal necrosis.Compared with model group,the neuronal damage of the rats in hippocampus regio in BMSCs group was alleviated,with restored morphology and improved neuronal loss.Compared with BMSCs group,the neurons of the rats in hippocampus region in combination group showed irregular morphology,disordered structure,unclear cell boundaries,uneven staining,and nuclear pyknosis.The Nissl staining results showed that the hippocampal neurons in sham operation group were tightly arranged with intact morphology,obvious nucleoli,and abundant darkly stained Nissl bodies.Compared with sham operation group,the neurons in hippocampus region of the rats in model group showed pyknosis,vacuolization,and sparse Nissl bodies.Compared with model group,the BMSCs group showed reduced neuronal pyknosis,relatively intact morphology,and increased Nissl bodies.Compared with BMSCs group,the combination group showed neuronal pyknosis,loss of morphological integrity,and fragmented Nissl bodies.The transmission electron microscope results showed that mitochondria in sham operation group exhibited oval shape with intact double-membrane structure and cristae.Compared with sham operation group,the mitochondria in model group showed swelling,disrupted membranes,broken cristae,and numerous autophagosomes.Compared with model group,the BMSCs group showed improved mitochondrial structure and reduced autophagosomes.Compared with BMSCs group,the combination group showed swollen mitochondria,disrupted membranes,broken cristae,and visible autophagosomes.The fluorescence probe results showed that compared with sham operation group,the ROS levels in the hippocampus neurons in brain tissue of the rats in model group were significantly increased(P<0.01);compared with model group,the ROS levels in hippocampus neurons in brain tissue of the rats in BMSCs group were significantly decreased(P<0.01);compared with BMSCs group,the ROS levels in hippocampus neurons in brain tissue of the rats in combination group were significantly increased(P<0.01).The Western blotting results showed that compared with sham operation group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in model group were significantly decreased(P<0.01);compared with model group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in BMSCs group were significantly increased(P<0.01);compared with BMSCs group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in combination group were significantly decreased(P<0.01);compared with sham operation group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,and LC3-Ⅱ/LC3-Ⅰ ratio of the rats in model group were significantly increased(P<0.01),while the expression level of P62 protein was significantly decreased(P<0.01);compared with model group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,as well as the LC3-Ⅱ/LC3-Ⅰ ratio,of the rats in BMSCs group were significantly decreased(P<0.01),while the expression level of P62 protein was significantly increased(P<0.01);compared with BMSCs group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,as well as the LC3-Ⅱ/LC3-Ⅰ ratio,of the rats in combination group were significantly increased(P<0.01),while the expression level of P62 protein was significantly decreased(P<0.01).Conclusion:BMSCs can alleviate the hippocampal neuronal pathological changes and improve cognitive function in the VaD rats,and its mechanism may be related to the regulation of ROS/Nrf2 signaling pathway to inhibit mitophagy.

Objective To observe the effects of Ditan Yizhi Decoction on learning and memory ability,structure of hippocampal tissue,neuronal morphology of hippocampus,and the expression of endoplasmic reticulum autophagy-related protein FAM134B in hippocampal tissue;To explore the mechanism of its therapeutic effect on vascular dementia.Methods Totally 32 SD rats were randomly divided into sham-operation group,model group,donepezil group and Ditan Yizhi Decoction group,with 8 rats in each group.The model group,donepezil group and Ditan Yizhi Decoction group were prepared with a modified permanent ligation method of bilateral common carotid arteries to create a rat model of vascular dementia,the common carotid artery was separated in the sham-operation group,but not ligated.After modeling,the donepezil group was given donepezil hydrochloride,Ditan Yizhi Decoction group was given Ditan Yizhi Decoction,and the sham-operation group and model group were given equal volume of distilled water for gavage for 4 consecutive weeks.Morris water maze experiment was used to evaluate the learning and memory ability,HE staining and Nissl staining were used to observe the morphological changes of hippocampus,ultrastructure of hippocampal neurons was observed using transmission electron microscopy,Western blot was used to detect the protein expression of FAM134B and p-FAM134B in hippocampal tissue.Results Compared with the sham-operation group,the escape latency period was prolonged of the rats in model group,and the number of crossing the original platform and the duration of stay in the target quadrant was reduced(P<0.01),the gap between neurons in CA1 region of the hippocampus increased,the cell morphology was irregular,the boundaries were blurred,the neurons shrinked,the Nissl bodies dissolved and broke,the number decreased,the endoplasmic reticulum arrangement was scattered,mitochondria swelled and deformed,and the expressions of FAM134B and p-FAM134B protein in hippocampal tissue increased(P<0.01).Compared with the model group,the escape latency period of rats in donepezil group and Ditan Yizhi Decoction group were significantly shortened,and the number of crossing the original platform and the duration of stay in the target quadrant were increased(P<0.01),the morphology and quantity of neurons in CA1 region of the hippocampus were more regular,with a decrease in neuronal pyknosis,an increase in the number of Nissl bodies,and a reduction in dissolution and fragmentation,the swelling and deformation of the endoplasmic reticulum were restored,and the expression of FAM134B and p-FAM134B protein in hippocampal tissue increased(P<0.01).Moreover,the effects of Ditan Yizhi Decoction group were better than those of the donepezil group(P<0.01).Conclusion Ditan Yizhi Decoction can improve the learning and memory ability and the morphology of neurons in vascular dementia rats.The mechanism may related to increasing the expression and phosphorylation of FAM134B protein,thereby promoting endoplasmic reticulum autophagy.

Objective:To investigate the clinical effect of esmolol on young and middle-aged patients with acute anterior myocardial infarction.Methods:Patients with acute anterior myocardial infarction from January 2008 to August 2020 were collected to obtain the basic information and clinical indicators. According to the clinical medication, the patients were divided into metoprolol group and esmolol group. The metoprolol group ( n=189) received routine esmolol, and the esmolol group ( n=104) received esmolol, intravenous injection, and then metoprolol sustained-release tablets. The clinical indexes, Gensini score, Killip grade, esmolol status and cardiac function after 7 d and 3 months of treatment were compared between the two groups. Results:Compared with the metoprolol group, the triglyceride (TG) was significantly higher, and the alanine aminotransferase (ALT) and aspartate aminotransferase (AST were significantly lower in the esmolol group (all P<0.05). The C-reactive protein (CRP), N-terminal pro-brain natriuretic peptide (NT-proBNP) and Gensini scores of culprit vessels in the esmolol group were lower than those in the metoprolol group ( P<0.05). There was no significant difference in cardiac function between the two groups within 7 d after treatment ( P>0.05). After 3 months of treatment, the left ventricular ejection fraction (LVEF) was higher and left ventricular end diastolic diameter (LVDD) was lower than those in the metoprolol group ( P<0.05). The number of postperative, ventricular tachycardia, shock and death in the esmolol group were lower than those in the metoprolol group, but the difference was not statistically significant ( P>0.05). Conclusions:Intravenous infusion of esmolol in young and middle-aged patients with acute anterior myocardial infarction can significantly improve the myocardial injury, liver function and cardiac function in prognosis.

Objective:To assess the changes in profiles of microRNAs (miRNAs) in peripheral blood of neonatal Sprague-Dawley (SD) rats with hypoxia-ischemia(HI) injury with self-resuscitation.Methods:Neonatal rats of 3 pregnant rats were divided into 3 groups according to their nests, in which group A was the blank control group, group B was the HI group, and group C was the alternative group.The expression profiles of miRNAs in periphe-ral blood of neonatal rats in group A and B by high-throughput sequencing was compared.Bioinformatics analysis was applied to investigate these differentially expressed miRNAs.Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to screen out enriched signaling pathways and functions.Target genes of miRNAs and those correlated with hypoxia-ischemia brain damage were predicted using miRBaseData (miRBD) software.HE staining was performed to observe the pathological changes of rat brain tissues.Results:A total of 1 049 mature reliable miRNAs in peripheral blood of neonatal rats were identified, including 525 miRNAs in group A, and 524 in group B. There were 27 differentially expressed miRNAs between group A and B, and their types were highly correlated.A total of 38 dysregulated miRNAs were screened out in group B, involving 21 upregulated miRNAs and 17 downregulated ones.GO and KEGG analyses showed that the identified differentially expressed miRNAs were mainly enriched in the glutamatergic synapse pathway, myelin lipid metabolism, neural activity ligand-receptor interaction and the vascular epithelial growth factor (VEGF) signaling pathway, all of them were significantly correlated with HIBD and over-activated.Cortex and subcallosal white matter lesions, enlarged ventricles, disordered arrangement of gray matter neurons, and obvious apoptosis in rat brain tissues of group A and B were not observed in HE staining.Conclusions:Differential expression of miRNAs in peripheral blood of HI self-resuscitated rats suggests that miRNAs has a positive response to hypoxia and ischemia.Differentially expressed miRNAs, including miR-200, miR-471, miR-429, miR-216 and miR-871 families, in peripheral blood of neonatal rat with HI showed their active response after HIBD.They are related to the molecular mechanisms of the nervous system damage, and are expected to become novel diagnostic markers for HIBD or HI.Differentially expressed miRNAs are conductive to the development of therapeutic targets of HI.

2019-novel coronavirus (2019-nCoV) is a highly pathogenic human CoV that first emerged in Wuhan in 2019.2019-nCoV has a zoonotic origin and poses a major threat to public health.However, little is known about the viral factors contributing to the high virulence of 2019-nCoV.Many animal viruses, including CoVs, encode proteins that interfere with host gene expression, including those involved in antiviral immune responses, and these viral proteins are often major virulence factors.Human coronaviruses (HCoVs) are known respiratory pathogens associated with a range of respiratory infection.In the past 17 years, the onset of 2019-nCoV, severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) have thrust HCoVs into spotlight of the research community due to their high pathogenicity in humans.The recent study of HCoVs-host interactions has contributed extensively to our understanding of infection pathogenesis of 2019-nCoV.This review discuss various host physiopathologic mechanism, such as apoptosis, innate immunity, endoplasmic reticulum (ER) stress response, mitogen-activated protein kinase (MAPK) pathway and nuclear factor kappa B (NF-κB) pathway that may be modulated by HCoVs and provides evidence for the intensive investigate of 2019-nCoV infection.

Objectve To investigate the application value of ultraifltration with millipore ifltration method test of old biological samples DNA. Methods 23 old blood samples separately were cut blood piece equally suitable size of three groups, labeled A, B and C group. DNA was extracted by magnetic bead methed and get DNA template with 80μL, 80μL, 20μL elution solution, respectively, then the DNA template in group A and C take right amount directly ampliifed, group B with milipore ultraifltration ifltration condensed amplication. PCR products were detected by ABI3130xl genetic analyzer and analyzed by GeneMapperID V3.2 software. The date were analyzed by using SPSS software. Results No samples were ampliifed for all STR loci in group A. There were 18 cases in group B samples amplified all STR loci and 11 samples in group C. Conclusion Application of millipore ultrafiltration method can signiifcantly improve the old biological samples DNA classiifcation success rate.

Objective:To explore the effect of different separation and purification technology on the physical and chemical proper-ties of water extract of ophiopogonin. Methods:The water extraction process of total ophiopogon saponins was optimized by an orthogo-nal test. The macroporous resin adsorption and membrane separation technology were adopted to purify the ophiopogonin water extract. The physical and chemical parameters, such as electrical conductivity, pH value, viscosity and turbidity, and the contents of total sap-onins, proteins, tannins and polysaccharides were determined. Results:The optimum water extraction technology of total saponins was as follows:6-fold amount of water was added, boiling 3 times with 90 min for each time. The electrical conductivity, pH value and vis-cosity of different purified liquid of total saponins had no significant differences, while the contents of total saponins and the three kinds of macromolecules showed significant differences. Conclusion: The content of macromolecules can be used as the reference index of purification process of total saponins water extract. Compared with macroporous resin separation, membrane separation technology is more suitable for the separation and purification of total saponins water extract.

To optimize the water extraction process for glycyrrhiza. Methods: HPLC was used to determine the con-tents of glycyrrhizic acid and liquiritin. The comprehensive index included the contents of glycyrrhizic acid and liquiritin and the yield of dry extract. The water amount and the extraction time were selected as the independent variables, and the comprehensive index was set as the dependent variable. Design-expert 8. 06 software was used to fit multivariate linear or quadratic multinomial models for the experimental values. Response surface methodology was used to optimize the water extraction process. The prediction was carried out through comparing the observed and predicted values. Results:The regression coefficient of binomial fitting complex model was as high as 0. 979 7. The optimum conditions of extraction process were as follows:12-fold amount of water, extracting 3 times with 90 min for each time. The deviation between the observed and predicted values was -1. 72%. Conclusion: Central composite design-response surface methodology is convenient and highly predictive in optimizing the water extraction process for glycyrrhiza, which can be applied in the further membrane separation and purification.

OBJECTIVETo study the molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation.

METHODThe cell proliferation was determined by MTT. The differentiation of the cells was detected by NBT reduction test. Cellular morphology was observed by Wright's staining. Cell cycle distribution and the distribution of CD11b, CD14 were detected by flow cytometry. Then RT-PCR and Western blot assay were employed to detect the expressions of c-myc, p27, CDK2 and cyclinE1 in HL-60 cells after exposure to TMP.

RESULTTMP inhibited the proliferation in a dose and time dependent manner. TMP at the concentration of 200 mg x L(-1) to 300 mg x L(-1) induced unterminal differentiation of HL-60 cell and synergistically blocked the cell cycle progression of HL-60 cells in G0/G1 phase. The expression of c-myc was down-regulated as well as the protein expression of cyclin E and CDK2, while the mRNA and protein expression of P27 were remarkably up-regulated.

CONCLUSIONSmall doses of TMP induces differentiation of HL-60 cells throughout the cell cyde, as detected by a slower rate of accumulation in G0/G1, possibly by regulating the expression and activity of G1/S phase-related molecules.

Cell Cycle Proteins ; genetics ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; genetics ; metabolism ; physiopathology ; Pyrazines ; pharmacology