Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.

Objective To observe whether bone marrow mesenchymal stem cell transplantation can improve vasospastic rats sense and motor function.Methods Rats grouped with randomized number method as Control group,Subarachnoid hemorrhage group.Stem cell culture media group and Stem cell transplantation group.Subarachnoid hemorrhage model were made with tail artery blood twice injection,2 days after 2' nd injection.Bone marrow mesenchymal stem cell were transplanted to lateral cistern.Subarachnoid hemorrhage(SAH) group didn' t transplant stem cell.Stem cell culture media group injected DMEM media as DMEM group.Stem cell transplantation group injected 30μl Bone Marrow mesenchymal stem cell suspension,so called BMSCs group.Neurofunctional score and learning memory expression were detected with morris mazer and Neurofunctional Score Scale in each group.Results After transplantation for 7 d,functional score of Control,SAH,DMEM and Stem cell group were 3.95 ±2.51,7.20 ± 1.03,7.23 ± 1.79 and 5.81 ± 1.11 respectively.Compared with others groups,Stem cell group score was significantly decrease(P=0.017).After transplanting stem cell for 14 d,the mean spanning plate time in Control group,SAH group,DMEM group and Stem cell group were 7.38 ± 1.73,4.52 ± 0.90,5.11 ± 1.93 and 7.32 ± 2.16 respectively,SAH and DMEM group vs other 2 groups,there were clearly statistically differences (P =0.009),while between control group and stem cell group,there were no statistically differences (P =0.14).Conclusion SAH rat transplant stem cell can improve sense,motor and learning expression in certain level.

[ ABSTRACT] AIM:To investigate the effect of artemisinin on lipopolysaccharide ( LPS)-induced intestinal epi-thelial barrier damage in IEC-6 cells and its molecular mechanism.METHODS:Cultured IEC-6 cells were divided to 5 groups:control group, LPS (100 mg/L) group and LPS +Artemisinin (30, 50 and 100μmol/L) groups.The cytotoxici-ty was detected by MTT assay.The releases of TNF-α, IL-1βand IL-6 in the IEC-6 cells were measured by ELISA.The transepithelial electrical resistance ( TER) was detected by electrical resistance tester, and the horseradish peroxidase (HRP) flux permeability were analyzed by a microplate reader.The expression of tight junction proteins, ZO-1, claudin-1 and occludin, and the expression of TLR4/MyD88/NF-κB at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:Artemisinin alone (up to 100 μmol/L) or in combination with LPS (100 mg/L) was not toxic to IEC-6 cells.Compared with control group, the releases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells significantly increased after treatment with LPS.The expression of TLR4/MyD88/NF-κB was activated by LPS.LPS down-regulated the protein expression of ZO-1, claudin-1 and occludin.However, artemisinin treatment decreased the re-leases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells.The expression of TLR4/MyD88/NF-κB at mR-NA and protein levels was gradually reduced after treatment with artemisinin.In addition, artemisinin upregulated the pro-tein expression of ZO-1, claudin-1 and occludin significantly (P<0.01) in a dose-dependent manner.CONCLUSION:Artemisinin attenuates LPS-induced intestinal epithelial barrier damage by inhibiting TLR4/MyD88/NF-κB activation in the IEC-6 cells.

AIM To explore the effects of glycyrrhetinic acid on the gastric ulcer rats infected by Helicobacter pylori (Hp) and its action mechanism.METHODS Gastric ulcer rat models were induced by acetic acid stress and then followed by Hp infection.After treatment with low and high doses of glycyrrhetinic acid,the ulcer index,gastric acid and proteinase activities in gastric ulcer rats were analyzed.The effects of glycyrrhetinic acid on the expressions of BCL2 and Caspase-3,the GSK3β activity in gastric mucosa and gastric epithelial cells,and the cell apoptosis level were then detected.RESULTS Glycyrrhetinic acid reduced the ulcer index,gastric acid and proteinase activities in rats.Besides,the expression of BCL2 was significantly up-regulated by glycyrrhetinic acid in gastric mucosa and gastric epithelial cells,whereas the expression of Caspase-3,level of cell apoptosis,and GSK3β activity were significantly reduced.After the treatment with GSK3 β activator LY294002,the level of BCL2 was down-regulated,Caspase-3 expression was increased,and the level of cell apoptosis was enhanced.CONCLUSION Glycyrrhetinic acid promotes the healing of gastric ulcer infected by Hp via regulating GSK3β activity and inhibiting apoptosis of gastric epithelial cells.

Practical ability training is one of the core tasks during the education for master of public health (MPH).We got a deeper understanding of practical ability cultivation situation for MPH in our school and got a more accurate recognition of the existed problems through conducting questionnaire for three grades MPH of professional degree and related tutors.We analyzed the advantages and disadvantages of the existing mode and reflected on curriculum design,teaching method reform and practical ability training reform,etc based on the results of the questionnaire and years of experiences.

Objective:To investigate the expressions of thioredoxin-interacting protein (TXNIP) /NLR family pyrin domain containing 3 (NLRP3) pathway in placental tissues of gestational diabetes mellitus (GDM) patients and its role in the migration and invasion of placental trophoblast cells.Methods:The placenta tissues of 25 GDM pregnant women (GDM group) and 25 normal glucose tolerance pregnant women (normal group) were collected. The cultured human chorionic trophoblast cells were divided into control group (untransfected) , siNC group (negative control) , siTXNIP group (siRNA-TXNIP) , siTXNIP+pcDNA group (co transfected with siRNA-TXNIP and pcDNA3.1 empty vector plasmids) and siTXNIP + NLRP3 group (co transfected with siRNA-TXNIP and pcDNA3.1-Flag-NLRP3 over expression plasmids) . The expressions of TXNIP and NLRP3 mRNAs were detected by real-time fluorescence quantitative PCR, and the expressions of TXNIP and NLRP3 proteins were detected by Western blot. Cell viability was measured by thiazole blue method. Cell migration ability was detected by scratch test, and cell invasion was detected by Transwell’s cell experiment.Results:Compared with those in the normal group, mRNA and protein expression levels of TXNIP and NLRP3 in the placenta of patients in GDM group were significantly higher ( P<0.05) . Compared with those in the control group or siNC group, mRNA and protein expression levels of TXNIP in siTXNIP group were significantly lower, however, the cell viability and migration rate were significantly higher, and the number of transmembrane cells was significantly higher ( P<0.05) ; but there was no significant difference in the above indexes between siNC group and the control group ( P>0.05) . Compared with those in siTXNIP group or siTXNIP + pcDNA group, mRNA and protein expression levels of NLRP3 in siTXNIP + NLRP3 group were significantly higher, however, the cell viability and migration rate were significantly lower, and the number of transmembrane cells was significantly lower ( P<0.05) ; but there was no significant difference in the above indexes between siTXNIP + pcDNA group and siTXNIP group ( P>0.05) . Conclusions:TXNIP/NLRP3 pathway is closely related to the occurrence and development of GDM. TXNIP can inhibit the migration and invasion of placental trophoblast cells by activating NLRP3.

To explore the potential mechanism of Qinfei Dayuan Granules for the treatment of pneumonia by the network pharmacology, the potential active ingredients and drug targets of Qinfei Dayuan Granules were obtained through the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP). The "component-target-disease" network and PPI network were constructed by Cytoscape 3.7.2 software, and GO functional enrichment analysis and KEGG pathway enrichment analysis were performed on the TCMIP platform to obtain a multi-dimensional network analysis of the "Chinese medicinal materials-chemical components-key targets-action pathways" and to explore the mechanism of its multi-component multi-target multi-action pathways of Qinfei Dayuan Granules for the treatment of pneumonia. A total of 474 active ingredients and 865 drug targets were identified from Qinfei Dayuan Granules; the key core targets of drugs include NF-κB, TNF-α, MAPK3, IL-1β, PTGS and CASP3, etc.. The results of GO functional enrichment analysis showed that drugs may interfere with inflammation through biological pathways such as immune regulation and apoptosis. KEGG signal pathway enrichment analysis showed that it was mainly related to the diabetic complications AGE-RAGE signaling pathway, IL-17 signaling pathway, T cell receptor signaling pathway and tumor necrosis factor signaling pathway, etc.. Qinfei Dayuan Granules can exert its effect on the treatment of pneumonia through inflammatory response and immune system with multi-ingredient, multi-target and multi-pathway pharmacological characteristics.

Objective:To analyze the detective value of placental tissue resistin, human lipid carrier protein (LCN) and blood glucose and lipid metabolism in pregnant women with gestational diabetes mellitus (GDM) complicated with preeclampsia (PE) , providing guidance for the early treatment of GDM complicated with preeclampsia.Methods:96 pregnant women with GDM complicated with PE (GDM-PE group) admitted to Yantai Yantaishan Hospital from Jan. 2017 to Jan. 2020 were selected and retrospectively studied. According to the ratio of 2:1, the pure GDM pregnant women (GDM group) and 48 normal pregnant women (the control group) were selected. The placenta tissue resistin and LCN levels were determined by immunohistochemistry. Blood samples were collected to determine the glucose and lipid metabolism. The pregnancy outcomes of each group were compared and the relationship between resistin, LCN, glucose and lipid metabolism and GDM complicated with PE was analyzed.Results:Fasting blood-glucose (FBG) was (4.57±0.66) mmol/L in GDM group and (5.23±0.61) mmol/L in GMD-PE group. FINS (11.97±1.5) mIU/L, (15.12±3.52) mIU/L were higher than those of control group (4.11±0.23) mmol/L, (6.75±1.34) mIU/L ( P<0.05) . FBG, FINS, glycosylated hemoglobin (HbA1c) in GDM-PE group were higher than those in GDM group. TC) (6.71±1.63) mmol/L, triglyceride, TG (6.59±0.87) mmol/L was higher than that of control group (5.87±0.73) mmol/L, (4.57±0.59) mmol/L and GDM group (6.02±1.55) mmol/L, (4.71±0.63) mmol/L ( P<0.05) . high density lipoprotein cholesterol (HDL-C) (1.21±0.34) was lower than that of control group (1.54±0.39) and GDM group (1.55±0.43) ( P<0.05) . The positive rates of resistin 85.42%, 60.42%, LCN 81.25%, 56.25% in GDM-PE group and GDM group were higher than those in control group 39.58%, 31.25% ( χ2=32.096, 4.167; 34.975, 6.095, both P<0.05) . The positive rates of resistin and LCN in GDM-PE group were higher than those in GDM group ( χ2=11.322, 11.257, both P<0.01) . The gestational age of delivery in GDM-PE group was (37.11±2.06) weeks earlier than that in GDM group (38.21±1.75) weeks and control group (38.36±1.42) weeks ( F=9.836, P<0.05) . The birth weight of neonates (2 905.45±356.79) g was lower than that of control group (3 321.52±366.46) g and GDM group (3 425.14±269.87) g ( F=46.606, P<0.05) . Postpartum blood loss (415.34±126.75) ml was significantly higher than that of GDM group (338.65±105.63) ml and control group (298.42±75.26) ml ( F=19.932, P<0.05) . The preterm birth rate of 20.83% was higher than that of the GDM group (8.33%) and the control group (4.17%) ( χ2=9.075, P<0.05) . The postpartum blood loss of the GDM group was higher than that of the control group ( t=-2.148, P<0.05) . The incidences of fetal distress, premature rupture of membranes, fetal growth restriction and postpartum hemorrhage in GDM-PE group were higher than those in control group ( χ2=4.571, 6.867, 5.941, 5.123, P<0.05) . The protein expressions of resistin and LCN in placenta of pregnant women with GDM-PE were positively correlated with FBG, FINS, TC and TG ( r=0.517, 0.463, 0.559, 0.521, 0.485, 0.497, 0.557, 0.571, P<0.05) . Was negatively correlated with HDL-C ( r=-0.317, -0.357, P<0.05) . Conclusions:The positive rate of resistin and LCN in the placenta tissue of pregnant women with GDM complicated with PE is higher than that of GDM and normal pregnant women, their disorder of glucose and lipid metabolism is more obvious, and the incidence of adverse maternal and infant outcomes is higher. It is speculated that resistin and LCN may synergistically affect the metabolism of glucose and lipids causing adverse pregnancy outcomes in GDM complicated with PE.

Objective:To investigate the changes of ROS/TXNIP/NLRP3 signaling pathway in pyroptosis of human embryonic trophoblast cells induced by high glucose.Methods:Human embryonic trophoblast cells were cultured in vitro to establish high glucose injury model, and they were randomly divided into control group, high glucose (HG) group and HG + ROS inhibitor N-acetyl-L-cysteine (HG + NAC) group. MTT assay was used to detect the cell survival rate. The level of ROS in each group was detected by dihydroethidine ROS fluorescence probe. Expression of TXNIP and NLRP3 mRNA was detected by real-time quantitative PCR (RT-qPCR). Western blot analysis was used to detect the expression levels of TXNIP, NLRP3, Caspase-1, interleukin (IL) -1β, tumor necrosis factor-α (TNF-α) and GSDMD proteins. In addition, pyroptosis was detected by flow cytometry.Results:The optimal glucose concentration for high glucose-induced injury of human embryonic trophoblast cells was 30 mmol/L. Compared with the control group (96.27±3.10) %, the survival rate of human embryonic trophoblast cells in HG group (55.44±2.15) % was significantly lower ( P<0.05), while the fluorescence intensity (ROS level) of 7 'dichlorofluorescein (DCF), the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly higher ( P<0.05) ; Compared with HG group, the survival rate of human embryonic trophoblast cells in HG+NAC group (84.75±2.33) % was significantly higher ( P<0.05), the fluorescence intensity (ROS level) of DCF, the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, and expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly lower ( P<0.05) . Conclusion:Inhibition of ROS level in human embryonic trophoblast cells induced by high glucose may promote cell proliferation and reduce the occurrence of pyroptosis by inhibiting TXNIP/NLRP3 signaling pathway.

OBJECTIVETo study the chemical constituents from the antioxidant fraction of Neo-Taraxacum siphonathum.

METHODVarious chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated on the basis of chemical evidence and spectral analysis.

RESULTTen compounds were isolated and identified from Neo-T. siphonathum, caffeic acid (1), chlorogenic acid (2), quercetin (3), luteolin (4), quercetin-3-O-beta-D-glucopyranoside (5), quercetin-3-O-alpha-D-arabinofuranoside (6), quercetin-3-O-alpha-D-arabinopyranoside (7), luteolin-7-O-beta-D-glucopyranoside (8), beta-sitosterol (9) and daucosterol (10).

CONCLUSIONCompounds 1-10 were isolated from this plant for the first time.

Caffeic Acids ; chemistry ; isolation & purification ; Chlorogenic Acid ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Luteolin ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Quercetin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Taraxacum ; chemistry