ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， the combination of serum pharmacochemistry， response profile of absorbed components in serum， network pharmacology and drug-likeness prediction was used to screen the potential active ingredients of Yanghetang against breast cancer. MethodsUPLC-Q-TOF-MS/MS was used to identify the main components in different solvent extracts of Yanghetang， and serum pharmacochemistry was applied to analyze the absorbed components from the serum of female SD rats after 0.5， 1， 2 h of administration. Combined with the response characteristic values of serum drug components obtained from UNIFI 1.8.2， the absorbed prototype components and metabolites were screened to get the absorbed components of Yanghetang with a significant patterns of elimination and growth. Network pharmacology was applied to construct a drug-component-pathway-target-disease network， and molecular docking was performed between absorbed components and key targets of breast cancer， and the drug similarity was analyzed by SwissADME. ResultsForty-two compounds were identified in Yanghetang samples extracted with different solvents， of which 16 compounds were common to the three different extraction solvents（methanol， 50% methanol and water）. The results of drug-containing serum analysis showed that there were 16 absorbed components in serum， including 5 prototypes and 11 metabolites. Network pharmacology results showed that Yanghetang against breast cancer involved 15 key targets such as proto-oncogene tyrosine-protein kinase Src（SRC）， epidermal growth factor receptor（EGFR） and phosphoinositide 3 kinase catalytic alpha polypeptide（PIK3CA）. Molecular docking results showed that 16 potential active ingredients were well combined with the predicted targets. Combined with drug likenesses， 12 compounds in the absorbed components of Yanghetang were considered to have potential for anti-breast cancer activity， mainly including α-pinene and γ-eudesmol and their metabolites， of which one was from Ephedrae Herba， one was from Rehmanniae Radix， and eight were from Cinnamomi Cortex. ConclusionThe chemical components of Yanghetang mainly include polysaccharides， monoterpene glycosides and coumarins， and its prototype components mainly undergo oxidation， hydrolysis and acetylation after entering the blood. Its anti-breast cancer mechanism may be related to the regulation of signaling pathways such as the mitogen-activated protein kinase（MAPK） and phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）. The results of this study can lay a foundation for further exploration of Yanghetang in the treatment of breast cancer.

OBJECTIVE: To observe the effect of electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) on hippocampal neuronal ferritinophagy mediated by the nuclear receptor coactivator 4 (NCOA4)/ferritin heavy chain 1 (FTH1) signaling pathway in vascular dementia (VD) rats, and to explore the potential mechanisms of electroacupuncture for VD. METHODS: A total of 60 male rats of SPF grade were randomly divided into a blank group (12 rats), a sham surgery group (12 rats) and a modeling group (36 rats). In the modeling group, the modified 4-vessel occlusion method was used to establish the VD model. The 24 successfully modeled rats were randomly divided into a model group and an electroacupuncture group, with 12 rats in each group. In the electroacupuncture group, electroacupuncture was applied at left and right "Sishencong" (EX-HN1), and bilateral "Fengchi" (GB20), with continuous wave, in frequency of 2 Hz and current intensity of 1 mA, 30 min a time, once daily for 21 consecutive days. The learning and memory abilities were assessed using the Morris water maze test before modeling, after modeling and after intervention, as well as the novel object recognition test after intervention. After intervention, the neuronal morphology in the hippocampus was observed by Nissl staining; the iron deposition was observed by Prussian blue staining; the reactive oxygen species (ROS) level was detected by dihydroethidium (DHE) fluorescence staining; the levels of iron, malondialdehyde (MDA) and superoxide dismutase (SOD) in the hippocampal tissue were measured by the colorimetric assay, TBA method, and WST-1 method, respectively; the positive expression of NCOA4, FTH1 and glutathione peroxidase 4 (GPX4) was detected by immunohistochemistry; the protein expression of NCOA4, FTH1, GPX4, and the ratio of microtubule-associated protein 1 light chain 3B (LC3B) Ⅱ/Ⅰ in the hippocampus were detected by Western blot. RESULTS: Compared with the sham surgery group, in the model group, the escape latency was prolonged, and the number of platform crossings reduced (P<0.01), the recognition index (RI) was decreased (P<0.01); the hippocampal neurons displayed a blurred laminar structure, disorganized cellular arrangement, and the number of Nissl bodies was decreased (P<0.01); the percentage of iron deposition area in the hippocampus was increased (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were increased (P<0.01), the SOD level, and the protein expression of FTH1 and GPX4 were decreased (P<0.01). Compared with the model group, in the electroacupuncture group, the escape latency was shortened and the number of platform crossings was increased (P<0.01), the RI was increased (P<0.01); the hippocampal neurons exhibited more regular morphology, better-organized cellular structure, and the number of Nissl bodies was increased (P<0.05); the percentage of iron deposition area in the hippocampus reduced (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were decreased (P<0.01, P<0.05), the SOD level, and the protein expression of FTH1 and GPX4 were increased (P<0.01). CONCLUSION Electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) can improve learning and memory abilities in VD rats, and its mechanism may be associated with the regulation of the hippocampal NCOA4/FTH1 signaling pathway, inhibition of ferritinophagy, and alleviation of oxidative stress damage.
Animals ; Electroacupuncture ; Dementia, Vascular/genetics* ; Male ; Rats ; Signal Transduction ; Humans ; Memory ; Rats, Sprague-Dawley ; Nuclear Receptor Coactivators/genetics* ; Ferritins/genetics* ; Learning ; Hippocampus/metabolism* ; Acupuncture Points

ObjectiveTo observe the effects of Xibining （XBN） and adipose stem cell exosome （ADSC-Exos） in the cases of separate or joint application on cartilage degeneration and mitochondrial autophagy and explore its mechanism of action to improve knee osteoarthritis （KOA）. MethodSD rats were divided into a sham operation group （sham group）， a model group， an ADSC-Exos group （Exos group）， an XBN group， and an ADSC-Exos+XBN group （Exos+XBN group）. KOA model was established by using anterior cruciate ligament transection （ACLT）. The pain sensitivity status of rats was evaluated， and the degeneration degree of the knee joint and cartilage tissue was detected by Micro-CT and pathological staining. The expression of p62 and LC3B was observed by immunofluorescence， and the serum levels of TNF-α， IL-1β， IL-6， and IL-15 in rats were detected by ELISA. The Western blot was used to detect the protein expression levels of MMP-3， MMP-13， ADAMTS5， ColⅡ， TIMP， ACAN， PINK1， Parkin， p62， and LC3A/B. ResultCompared with the sham group， rats in the model group showed decreased cold-stimulated foot-shrinkage thresholds and mechanical pain sensitivity thresholds， varying degrees of abrasion and loss of cartilage tissue， degeneration of cartilage tissue， elevated serum IL-1β， IL-6， IL-15， and TNF-α levels （P<0.01）， and increased protein expression of MMP-3， MMP-13， and ADAMTS5 in cartilage tissue. In addition， the protein expression of ColⅡ， TIMP1， and ACAN was decreased （P<0.01）. Compared with the model group， rats in each treatment group showed higher cold-stimulated foot-shrinkage thresholds and mechanical pain sensitivity thresholds， reduced cartilage tissue degeneration， lower serum levels of IL-1β， IL-6， IL-15， and TNF-α （P<0.05，P<0.01）， decreased protein expression of MMP-3， MMP-13， and ADAMTS5， and higher protein expression of Cold， TIMP1， and ACAN in cartilage tissue （P<0.05，P<0.01）. Moreover， the changes were the most obvious in the Exos+XBN group. ConclusionBoth ADSCs-Exos and XBN can increase the level of mitochondrial autophagy in chondrocytes and delay cartilage tissue degeneration by promoting the expression of the PINK1/Parkin signaling pathway， and the combination of the two can enhance the therapeutic effect.

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) on learning and memory impairment in vascular dementia (VD) rats by observing the influences on the N-methyl-D-aspartate receptor (NMDAR)/cyclic adenosine monophosphate response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway and the excitotoxicity induced by hippocampal calcium overload. METHODS: Thirty-two male SD rats of SPF grade were selected and randomized into a normal group (6 rats), a sham-operation group (6 rats) and an operation group (20 rats). VD model was established with the modified Pulsinelli's four-vessel occlusion (4-VO) method. Twelve rats after successfully modeled were assigned randomly into a model group and an EA group, 6 rats in each one. In the EA group, EA was delivered at bilateral "Fengchi" (GB 20) and "Sishencong" (EX-HN 1), with the continuous wave, the frequency of 2 Hz and the electric current of 1 mA. Stimulation intensity was adjusted depending on the slightly trembling of rat head. EA was given once daily, 30 min each time; and EA intervention was delivered for 21 days continuously. Using Morris water maze test, the learning and memory function was assessed. The neuronal morphology in the hippocampal CA1 was observed with HE staining; the level of glutamate (GLU) in serum and hippocampal tissue, as well as the activity of calcium pump (Ca²⁺-ATP) in the hippocampus were detected using colorimetric method. The protein expression of NMDAR, calmodulin-dependent protein kinase Ⅱ (CaMKⅡ), phosphorylated calmodulin-dependent protein kinase Ⅱ (p-CaMKⅡ), phosphorylated cyclic phosphoradenosine effector element binding proteins (p-CREB), CREB, and BDNF in the hippocampal CA1 was detected using immunohistochemistry. The protein expression of NMDAR, CREB, p-CREB and BDNF in the hippocampal tissue was detected using Western blot method. RESULTS: Compared to the sham-operation group, in the model group, the escape latency was prolonged and the platform crossing times of rats were reduced (P<0.01), the hippocampal neuron structure was damaged to different degrees, the structure in hippocampal CA1 was loosened, the arrangement disorganized, with clear grid-like structure; the neuronal morphology was irregular, pyknosis and even dissolution occurred, glial cells increased, blood capillary was dilated and the inflammatory cells were infiltrated and scattered. The level of GLU in the serum and hippocampal tissue and the protein expression of hippocampal NMDAR were elevated (P<0.01), the activity of Ca²⁺-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were reduced (P<0.01, P<0.05); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB were dropped (P<0.05). In comparison with the model group, in the EA group, the escape latency was shortened and the platform crossing times of rats rose (P<0.01), the arrangement was improved in the hippocampal CA1, the neuronal morphology was intact, the nucleoli were clear relatively and the pyknosis or dissolution were attenuated, the numbers of glial cells reduced relatively, the dilation of blood capillary was alleviated. The level of GLU in the serum and hippocampal tissue and the protein expression of NMDAR were reduced in the hippocampal tissue (P<0.01), the activity of Ca²⁺-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were elevated (P<0.05, P<0.01); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB increased (P<0.05). CONCLUSION EA at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) can attenuate learning and memory impairment in VD rats, which may be obtained by reducing GLU level in hippocampal tissue, inhibiting hippocampal excitotoxicity, mediating protein expression related to the NMDAR/CREB/BDNF signaling pathway, and maintaining neuronal survival and growth.
Electroacupuncture ; Male ; Animals ; Rats, Sprague-Dawley ; Learning ; Memory ; Signal Transduction ; Cyclic AMP Response Element-Binding Protein/metabolism* ; Memory Disorders/therapy* ; Brain-Derived Neurotrophic Factor/metabolism* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Dementia, Vascular/therapy*

Objective To analyze the predictive value of serum soluble intercellular adhesion molecule-1(sICAM-1),soluble vascular intercellular adhesion molecule-1(sVCAM-1)and soluble microfiber associated protein 4(sMFAP4)for major adverse cardiovascular events(MACE)after per-cutaneous coronary intervention(PCI)in acute myocardial infarction.Methods A total of 166 pa-tients with acute myocardial infarction after PCI were included in case group,and 166 healthy volun-teers were included in control group during the same period.Enzyme-linked immunosorbent assay was used to detect serum levels of sICAM-1,sVCAM-1 and sMFAP4,and the differences between groups were compared.After being followed-up for 1 year,the case group was divided into MACE group(n=42)and non-MACE(n=113)group after removing 10 cases.General information was compared be-tween the MACE group and the non-MACE group.Logistic regression analysis was used to identify the influencing factors for MACE;receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was used to evaluate the efficacy of relevant indicators in predicting MACE.Results The incidence of MACE was 27.10％.The serum levels of sICAM-1,sVCAM-1 and sMFAP4 in the case group were significantly higher than those in the control group,and those in the MACE group were significantly higher than those in the non-MACE group(P＜0.05).Smoking history(OR=3.688,95％CI,1.107 to 12.286),drinking history(OR=3.364,95％CI,1.238 to 9.139),combined hypertension(OR=4.255,95％CI,1.250 to 14.483),combined type 2 di-abetes(OR=4.208,95％CI,1.051 to 16.856),combined hyperlipidemia(OR=5.238,95％CI,1.440 to 19.061),Gensini score(OR=5.579,95％CI,1.355 to 22.968),blood platelet count(PLT)(OR=0.519,95％CI,0.281 to 0.961),serum sICAM-1(OR=5.013,95％CI,1.859 to 13.514),sVCAM-1(OR=4.826,95％CI,1.769 to 13.164)as well as sMFAP4(OR=4.745,95％CI,1.372 to 16.407)and no reflow during surgery(OR=2.962,95％CI,1.107 to 7.924),no collateral circulation formation(OR=3.225,95％CI,1.173 to 8.867)were all influen-cing factors for the occurrence of MACE.Serum sICAM-1,sVCAM-1 and sMFAP4 levels predicted the cut-off value of MACE in the case group were 331.53 ng/mL,473.20 ng/mL and 30.63 U/L,respectively,the sensitivity was 80.95％,76.19％and 78.57％,respectively,the specificity was 94.69％,97.35％,88.50％,respectively,and the AUC was 0.835(95％CI,0.767 to 0.890),0.794(95％CI,0.722 to 0.855),0.824(95％CI,0.754 to 0.880),respectively.The sensitivi-ty,specificity and AUC of serum sICAM-1,sVCAM-1 and sMFAP4 to predict MACE were larger or higher than those of Gensini score(P＜0.05);the sensitivity,specificity and AUC of the above ser-um indexes combined to predict the occurrence of MACE in the case group were 100.00％,94.69％and 0.956(95％CI,0.910 to 0.982),respectively,and the sensitivity and AUC of the combined prediction were higher or larger than those of the single prediction(P＜0.05).Conclusion Serum sICAM-1,sVCAM-1 and sMFAP4 levels are increased in patients with acute myocardial infarction,all of which are related to MACE after PCI.The MACE prediction value of the three is better than Gensini score,and the combined prediction value of the three is higher.

Objective To analyze the predictive value of serum soluble intercellular adhesion molecule-1(sICAM-1),soluble vascular intercellular adhesion molecule-1(sVCAM-1)and soluble microfiber associated protein 4(sMFAP4)for major adverse cardiovascular events(MACE)after per-cutaneous coronary intervention(PCI)in acute myocardial infarction.Methods A total of 166 pa-tients with acute myocardial infarction after PCI were included in case group,and 166 healthy volun-teers were included in control group during the same period.Enzyme-linked immunosorbent assay was used to detect serum levels of sICAM-1,sVCAM-1 and sMFAP4,and the differences between groups were compared.After being followed-up for 1 year,the case group was divided into MACE group(n=42)and non-MACE(n=113)group after removing 10 cases.General information was compared be-tween the MACE group and the non-MACE group.Logistic regression analysis was used to identify the influencing factors for MACE;receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was used to evaluate the efficacy of relevant indicators in predicting MACE.Results The incidence of MACE was 27.10％.The serum levels of sICAM-1,sVCAM-1 and sMFAP4 in the case group were significantly higher than those in the control group,and those in the MACE group were significantly higher than those in the non-MACE group(P＜0.05).Smoking history(OR=3.688,95％CI,1.107 to 12.286),drinking history(OR=3.364,95％CI,1.238 to 9.139),combined hypertension(OR=4.255,95％CI,1.250 to 14.483),combined type 2 di-abetes(OR=4.208,95％CI,1.051 to 16.856),combined hyperlipidemia(OR=5.238,95％CI,1.440 to 19.061),Gensini score(OR=5.579,95％CI,1.355 to 22.968),blood platelet count(PLT)(OR=0.519,95％CI,0.281 to 0.961),serum sICAM-1(OR=5.013,95％CI,1.859 to 13.514),sVCAM-1(OR=4.826,95％CI,1.769 to 13.164)as well as sMFAP4(OR=4.745,95％CI,1.372 to 16.407)and no reflow during surgery(OR=2.962,95％CI,1.107 to 7.924),no collateral circulation formation(OR=3.225,95％CI,1.173 to 8.867)were all influen-cing factors for the occurrence of MACE.Serum sICAM-1,sVCAM-1 and sMFAP4 levels predicted the cut-off value of MACE in the case group were 331.53 ng/mL,473.20 ng/mL and 30.63 U/L,respectively,the sensitivity was 80.95％,76.19％and 78.57％,respectively,the specificity was 94.69％,97.35％,88.50％,respectively,and the AUC was 0.835(95％CI,0.767 to 0.890),0.794(95％CI,0.722 to 0.855),0.824(95％CI,0.754 to 0.880),respectively.The sensitivi-ty,specificity and AUC of serum sICAM-1,sVCAM-1 and sMFAP4 to predict MACE were larger or higher than those of Gensini score(P＜0.05);the sensitivity,specificity and AUC of the above ser-um indexes combined to predict the occurrence of MACE in the case group were 100.00％,94.69％and 0.956(95％CI,0.910 to 0.982),respectively,and the sensitivity and AUC of the combined prediction were higher or larger than those of the single prediction(P＜0.05).Conclusion Serum sICAM-1,sVCAM-1 and sMFAP4 levels are increased in patients with acute myocardial infarction,all of which are related to MACE after PCI.The MACE prediction value of the three is better than Gensini score,and the combined prediction value of the three is higher.

Understanding the regulatory networks for germ cell fate specification is necessary to developing strategies for improving the efficiency of germ cell production in vitro. In this study, we developed a coupled screening strategy that took advantage of an arrayed bi-molecular fluorescence complementation (BiFC) platform for protein-protein interaction screens and epiblast-like cell (EpiLC)-induction assays using reporter mouse embryonic stem cells (mESCs). Investigation of candidate interaction partners of core human pluripotent factors OCT4, NANOG, KLF4 and SOX2 in EpiLC differentiation assays identified novel primordial germ cell (PGC)-inducing factors including BEN-domain (BEND/Bend) family members. Through RNA-seq, ChIP-seq, and ATAC-seq analyses, we showed that Bend5 worked together with Bend4 and helped mark chromatin boundaries to promote EpiLC induction in vitro. Our findings suggest that BEND/Bend proteins represent a new family of transcriptional modulators and chromatin boundary factors that participate in gene expression regulation during early germline development.
Animals ; Cell Differentiation/genetics* ; Chromatin/metabolism* ; Embryonic Stem Cells ; Germ Cells/metabolism* ; Germ Layers/metabolism* ; Mice

Objective: To apply the semi-quantitative risk assessment model in the Technical Guidelines for Occupational Health Risk Assessment of Chemical Hazardous Factors in the Workplace to assess the occupational health risk of electroplating enterprises and explore its applicable conditions and characteristics. Methods: Three electroplating enterprises were selected as the research objects. Occupational hygiene survey and hazard factor detection were conducted on the spot. Three semi-quantitative risk assessment methods were used to assess the risk, and the evaluation results were compared. Results: The consistency between the contact index method and the comprehensive index method was strong. The weighted Kappa value was 0.946 and the P value was less than 0.001, but the consistency between the contact ratio method, the contact index method and the comprehensive index method was poor. The weighted Kappa value was 0.345 and 0.391, and the P value was 0.009 and 0.004, respectively. When the contact concentration is less than 50% OELs, the evaluation results of the contact ratio method are lower than those of the exponential method and the comprehensive exponential method. The consistency of the three methods was the highest when the contact concentration (>50%) and (OELs, the results of the contact ratio method are higher than those of the index method and the comprehensive index method. Conclusion Contact ratio method is suitable for occupational health risk assessment under the condition of incomplete occupational health information and for enterprise managers to identify key control points of health risk through self-assessment; Contact index method is suitable for hazard risk assessment of occupational hazards without sampling test conditions or OELs, and pre-assessment of occupational hazards of construction projects without access to analogical test data. The index method is suitable for occupational health risk assessment with available testing data and complete occupational health information.

Objective This study aims to investigate the potential correlation between the rs1187929 polymorphism of tyrosine kinase B gene and post-stroke depression (PSD). Methods A total of 78 depression patients (PSD) and 112 non-depression patient controls (NPSD patients) were included in the study. All participants were evaluated using the Hamilton Rating Scale for Depression (HAMD) and further divided into PSD and NPSD groups. PSD patients were diagnosed in accordance with DSM-V criteria. The rs1187929 polymorphism of TrkB was genotyped by using PCR product purification and Sanger double termination sequencing. Results The total prevalence rate was 41.05% . The prevalence rate was 42.86% in men and 38.82% in women ( 字2=0.316,P=0.57). The CC, CT and TT frequencies of PSD group were 26.92% , 48.72% and 24.36% respectively. The CC, CT and TT frequencies of NPSD group were 42.86%, 45.54% and 11.61% respectively. Based on the CC genotype, the relative risk of homozygous mutant TT was 3.341 (字2=7.347, P=0.007, OR=3.341),and the T allele frequency in the PSD group was 48.72% which was higher than 34.38% in the NPSD group. The relative risk of allele T relative to allele C was 1.814 (字2=7.798, P=0.005, OR=1.814). Homozygous mutant TT and allele T were risk factors for post-stroke depression (P<0.05). Conclusion There is a correlation between rs1778929 gene polymorphism and PSD in Han population in South Anhui Province, which warrants further large-scale studies.