Berberine Inhibits Atherosclerosis by Regulating Lipophagy Via Targeting Wnt5a/NPC1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect and molecular mechanism of berberine （BBR） on lipophagy in the prevention and treatment of atherosclerotic （AS） lesions in mice. MethodFifty apolipoprotein E-knockout （ApoE-/-） mice were randomly divided into an AS model group， an atorvastatin group （5 mg·kg-1）， and low-， medium-， and high-dose BBR groups （2.5， 5， 10 mg·kg-1）. Ten C57BL/6J mice were assigned to the control group. After 12 weeks， hematoxylin-eosin （HE） and oil red O staining were performed to assess the histopathological changes of AS plaques in the aorta. Biochemical analysis was used to measure serum lipid levels， and enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of inflammatory cytokines interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）， oxidative stress marker reactive oxygen species （ROS）， and serum lipophagy marker Beclin1 and microtubule-associated protein 1 light chain 3 Ⅱ （LC3Ⅱ）. The xanthine oxidase method was used to measure serum superoxide dismutase （SOD） activity. Immunohistochemistry （IHC） was used to detect the distribution of wingless-type MMTV integration site family member 5a （Wnt5a） and Nieman Pick type C1 （NPC1） in the aorta， and Western blot was used to determine the protein expression of Wnt5a and NPC1 in the aorta. ResultCompared with the control group， the AS model group showed significant AS plaque formation， significantly elevated levels of serum total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein cholesterol （LDL-C）， IL-6， TNF-α， and ROS， aortic Wnt5a distribution and protein expression （P<0.01）， and significantly reduced levels of serum high-density lipoprotein cholesterol （HDL-C）， SOD， Beclin1， LC3Ⅱ， and aortic NPC1 distribution and protein expression （P<0.01）. Compared with the AS model group， the atorvastatin group， and high- and medium-dose BBR groups showed a significant reduction in AS plaque area （P<0.05， P<0.01）， significantly decreased levels of serum TC， TG， LDL-C， IL-6， TNF-α， ROS， and aortic Wnt5a distribution and protein expression （P<0.05， P<0.01）， and significantly increased levels of serum HDL-C， SOD， Beclin1， LC3Ⅱ， and aortic NPC1 distribution and protein expression （P<0.05， P<0.01）. There was no statistically significant difference in the above indicators between the atorvastatin group and the medium-dose BBR group. ConclusionBBR can competitively bind to Wnt5a to activate NPC1 expression， upregulate lipophagy levels， reduce blood lipids， and inhibit the release of inflammatory mediators and oxidative stress damage， thereby exerting a preventive and therapeutic effect on AS.

Anemoside B4 regulates fatty acid metabolism reprogramming in mice with colitis-associated cancer / 中国中药杂志

The study aimed to investigate the effect of anemoside B4(B4) on fatty acid metabolism in mice with colitis-associated cancer(CAC). The CAC model was established by azoxymethane(AOM)/dextran sodium sulfate(DSS) in mice. Mice were randomly divided into a normal group, a model group, and low-, medium-, and high-dose anemoside B4 groups. After the experiment, the length of the mouse colon and the size of the tumor were measured, and the pathological alterations in the mouse colon were observed using hematoxylin-eosin(HE) staining. The slices of the colon tumor were obtained for spatial metabolome analysis to analyze the distribution of fatty acid metabolism-related substances in the tumor. The mRNA levels of SREBP-1, FAS, ACCα, SCD-1, PPARα, ACOX, UCP-2, and CPT-1 were determined by real-time quantitative PCR(RT-qPCR). The results revealed that the model group showed decreased body weight(P<0.05) and colon length(P<0.001), increased number of tumors, and increased pathological score(P<0.01). Spatial metabolome analysis revealed that the content of fatty acids and their derivatives, carnitine, and phospholipid in the colon tumor was increased. RT-qPCR results indicated that fatty acid de novo synthesis and β-oxidation-related genes, such as SREBP-1, FASN, ACCα, SCD-1, ACOX, UCP-2, and CPT-1 mRNA expression levels increased considerably(P<0.05, P<0.001). After anemoside B4 administration, the colon length increased(P<0.01), and the number of tumors decreased in the high-dose anemoside B4 group(P<0.05). Additionally, spatial metabolome analysis showed that anemoside B4 could decrease the content of fatty acids and their derivatives, carnitine, and phospholipids in colon tumors. Meanwhile, anemoside B4 could also down-regulate the expression of FASN, ACCα, SCD-1, PPARα, ACOX, UCP-2, and CPT-1 in the colon(P<0.05, P<0.01, P<0.001). The findings of this study show that anemoside B4 may inhibit CAC via regulating fatty acid metabolism reprogramming.

Prediction of the consistency of large pituitary adenoma based on CT density combined with texture parameter modeling / 中国医学影像技术

Objective: To explore the value of CT density combined with texture parameters based on CT plain image in predicting the consistency of large pituitary adenoma. Methods: Totally 50 patients with large pituitary adenoma confirmed by operation and pathology were enrolled and divided into soft group (n=30) and hard group (n=20) according to intraoperative pituitary consistency. The largest slice of the tumor on the CT image was selected, then ROI was manually outlined, CT value of the lesion was measured, and the texture feature parameters were extracted. CT values and texture features were compared between the two groups. Multivariate Logistic regression analysis was used to analyze the variables, and the model for predicting the pituitary adenoma consistency was established. ROC curve was drawn to evaluate its predictive value. Results: There was statistically significant difference in CT value between soft group and the hard group (P=0.031), and AUC in predicting tumor consistency was 0.662. A total of 77 texture parameters were extracted based on plain CT images, and 4 texture parameters were found with statistically significant differences between the two groups, including the Quantile 90, inertia, variance and contrast, with AUC of 0.662, 0.663, 0.672 and 0.663, respectively. AUC of texture feature model established with multivariate Logistic regression analysis in predicting the pituitary adenoma consistency was 0.690, of CT value combined with the texture parameter model was 0.782. Conclusion: The model established with CT value combined with texture parameters has high value in predicting the pituitary adenoma consistency, which is helpful to clinical selection of surgical plans.

Comparison of PET/CT performance after oral administration and intravenous injection of 18F-NaF / 中华核医学与分子影像杂志

Objective@#To investigate the possibility and diagnostic efficiency of 18F-NaF PET/CT bone scan after oral administration (PO) by comparing with that of intravenous injection (IV).@*Methods@#Fifty patients (19 males, 31 females; average age: (52.8±11.7) years) with cancer who underwent PET/CT scans after oral and intravenous administration of 18F-NaF respectively with an interval of 2-7 d from June 2015 to September 2016 were prospectively enrolled. Single-phase 18F-NaF PET/CT was performed 60 min after IV, and dual-phase 18F-NaF PET/CT was performed 60 and 120 min after PO. All PET/CT images were reviewed, lesions were counted, and maximum standardized uptake value (SUVmax) and target/non-target (T/NT) ratios were calculated and compared. Paired t test was used.@*Results@#Forty-one patients (15 males, 26 females; average age: (53.5±10.4) years) who finished all PET/CT scans were enrolled. The images at 120 min after PO was visually similar to the images at 60 min after IV. Three modalities detected the same cases and lesions (35 positive cases: 25 malignant, 8 benign, 2 cases with indefinite diagnosis; 302 lesions: 172 malignant, 108 benign, 22 ambiguous lesions). The SUVmax-PO60 min and SUVmax-PO120 min were lower than the SUVmax-IV60 min in the same lesion (18.22±12.64, 26.60±19.49 vs 28.07±16.34; t values: -12.36 and -3.59, both P<0.05). A total of 194 lesions were included for T/NT ratio analysis. T/NTIV60 min, T/NTPO60 min and T/NTPO120 min were 2.76±1.30, 2.87±1.50, 2.98±1.42, respectively, and T/NTPO120 min was higher than T/NTIV60 min (t=3.18, P<0.05).@*Conclusion@#18F-NaF PET/CT images after PO, especially at 120 min post-PO, has similar diagnostic power of lesion-detection and SUVmax-measurement with IV.

Tetramethylpyrazine inhibits PM2.5-induced vascular smooth muscle cell proliferation by down-regulating JNK phosphorylation / 中国药理学与毒理学杂志

OBJECTIVE To investigate the inhibitory effect and the possible mechanism of tetra-methylpyrazine (TMP) in preventing vascular smooth muscle cells (VSMCs) proliferation induced by fine particulate matter (PM2.5). METHODS PM2.520, 200 and 400 mg · L-1 was added to VSMCs for 24 h, the survival of VSMCs was measured by MTT assay, the protein levels of p-c-Jun N-terminal kinase (JNK) and fibroblast growth factor receptor-1 (FGFR-1) in the VSMCs were detected by Western blotting, while the levels of vascular cell adhesion molecule-1 (VCAM-1), endothelin-1 (ET-1) and nitric oxide (NO) in the VSMCs were analyzed by ELISA, radioimmunoassay and nitrate reductase method, respec-tively. TMP 20, 200 and 2000 mg·L-1 or a specific inhibitor of JNK SP60012510μmol·L-1 was added into the VSMCs to observe the effect of TMP. RESULTS Compared with the normal control group, PM2.5200 and 400 mg·L-1 significantly increased the A570 nm vaule, the protein levels of p-JNK and FGFR-1,the levels of VCAM-1 and ET-1, but decreased the level of NO (P<0.01), while there were no significant changes in PM2.520 mg·L-1 group. Compared with the PM2.5200 mg·L-1 group, TMP 200 and 2000 mg·L-1 pre-treatment markedly decreased the A570 nm vaule, the protein levels of p-JNK and FGFR-1, the levels of VCAM-1 and ET-1, but increased the level of NO (P<0.01), while there were no significant changes in TMP 20 mg · L-1 pre-treated group. Moreover, the effects of TMP were significantly enhanced by the co-incubation of TMP 2000 mg · L-1 with SP60012510 μmol · L-1, compared to the TMP 2000 mg · L-1 pre-treated group (P<0.05, P<0.01). CONCLUSION TMP displays a significant inhibitory effect against VSMC proliferation induced by PM2.5. The mechanism may be related to the inhibition of JNK phosphor-ylation, and the regulation of FGFR-1 protein expression and VCAM-1, ET-1 and NO levels.

Establishment of a mouse model of IgA nephropathy induced by superantigen staphylococcal enterotoxin B / 中国比较医学杂志

Objective To establish a mouse model of IgA nephropathy and to observe its biochemical and pathological characteristics. Methods Twelve BALB/c mice were randomly divided into the normal group and model group, with 6 mice in each group. Mice in the model group received an intravenous injection of 0. 8 mg/kg superantigen staphylococcal enterotoxin B (SEB) into the tail vein once a week for three weeks. At the end of the 4th week, the mice were sacrificed, and the 24 h-urinary protein, urinary microalbumin, the renal function indicators BUN, Scr and UA were measured, levels of liver function indicators ALT, AST, ALP, and the blood lipid levels of TC, TG, and LDL were determined, the renal morphological changes were examined by pathology using HE, PAS, PASM and Masson staining, and by electron microscopy, the IgA deposition in the renal tissue was observed with immunofluorescence, and the liver and small intestine were observed by pathology using HE staining. Results Compared with the normal group, the mice of model group showed increased 24-hour urinary protein and urinary microalbumin (P<0. 01), increased CREA and UA (P<0. 05), but not significantly changed BUN, TP and ALB. The liver function indicator AST was significantly increased (P<0. 05), but ALT and ALP were not significantly changed. The blood lipid TG was significantly decreased (P<0. 05) and LDL increased (P<0. 01), while the TC was not significantly changed. The kidney tissues had moderate histological changes, and immunofluorescence observation showed granular or massive IgA deposition in the renal glomerular mesangium. The liver tissue had some inflammatory cell infiltration and hepatocyte necrosis. The small intestine showed slender and shortened villi with widened inter-villous space and sloughed off epithelial cells, dilated central lacteal, and lymphocyte infiltration. Conclusions A mouse model of IgA nephropathy can be successfully established by tail vein injection of superantigen staphylococcal entrotoxin B.

Effect of berberine on Helicobacter pylori-induced human gastric epithe-lial cells injury / 中国病理生理杂志

AIM: To investigate the effect of berberine (Ber) on Helicobacter pylori (Hp)-induced human gastric epithelial cells (GES-1) injury and the underlying mechanism .METHODS: Berberine (5, 10 and 20 μmol/L) and PD98059 (20 μmol/L), a selective inhibitor of extracellular regulated protein kinases (ERK)1/2 signaling pathway, were added to Hp-infected GES-1 cells.The cell activity and apoptosis, the levels of interleukin (IL)-1βand IL-8, lactic dehydrogenase ( LDH) activity and the protein levels of Bax , Bcl-2 and p-ERK1/2 in the GES-1 cells were determined by MTT assay, flow cytometry, ELISA, colorimetry and Western blot, respectively.RESULTS: Compared with control group, Hp significantly inhibited the cell activity , increased the apoptotic rate , LDH activity, IL-1βand IL-8 levels, the Bax and p-ERK1/2 protein levels but decreased the Bcl-2 protein level in GES-1 cells (P<0.05).However, these effects of Hp were reversed by berberine at medium-dose and high-dose, as compared with the Hp-infected GES-1 cells ( P<0.05).Moreover, the protective effects of berberine were significantly enhanced by the co-incubation of berberine with PD98059, as compared with the berberine at higher dose (P<0.05).CONCLUSION:Berberine may attenuate Hp-in-duced human gastric epithelial GES-1 cells injury by anti-inflammation, promoting cell growth and anti-apoptosis via the in-hibition of ERK1/2 signaling pathway .

Puerarin attenuates PM2.5-induced vascular endothelial cells injury via ERK1/2 signaling pathway / 中国中药杂志

To investigate the effect and the mechanism of puerarin in attenuating PM2.5-induced human umbilical vein endothelial cells (EA.hy926) injury, the samples of fine particulate matter (PM2.5) were collected and made into suspension. Different concentrations of PM2.5 (0,20, 200, 400 mg•L⁻¹) were used to contaminate EA.hy926 cells for 24 h. The cells survival rate was detected by MTT assay; cells apoptosis of EA.hy926cells was detected by flow cytometry; the protein levels of p-ERK1/2, Bax and Bcl-2 were detected by Western blot; the contents of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malonaldehyde (MDA), and the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) were measured by ELISA. Puerarin at different concentrations (10, 50, 100 μmol•L⁻¹) or a specific inhibitor of ERK1/2 pathway PD98059 (20 μmol•L-1) was added into the EA.hy926 cells to observe the intervention effect and mechanism of puerarin. Compared with the control group, PM2.5 reduced the cells survival rate, up-regulatedp-ERK1/2 protein level and Bax/Bcl-2 ratio in a dose dependent manner to promote apoptosis; increased the contents of TNF-α, IL-6 and MDA, the activity of LDH, but decreased SOD activity in the EA.hy926 cells (P<0.05). Compared with PM2.5 group, puerarin increased the cells survival rate, down-regulated p-ERK1/2 protein level and Bax/Bcl-2 ratio in a dose dependent manner to inhibit the apoptosis; decreased the contents of TNF-α, IL-6 and MDA, the activity of LDH, but increased SOD activity in the EA.hy926 cells (P<0.05). The results indicated that puerarin could attenuate PM2.5-induced EA.hy926 cells injury via the inhibition of ERK1/2 pathway.

Synthesis and antitumor activity of cinnamoyloxy phosphonate derivatives / 药学学报

In search of more effective anticancer agents, twelve compounds were designed and synthesized via microwave-assisted reactions of cinnamoyl chloride with α-hydroxyphosphonate. The structures of all the compounds were confirmed by IR, NMR and elemental analysis. Bioassay of the compounds were tested. They exhibited certain antitumor activities. Especially, compound 3c had obvious inhibitory effect on growth of SGC-7901 cells in vitro at 20 μmol·L-1, and compound 3h showed better inhibitory effect on growth of SGC-7901 cells in vitro at 5 μmol·L-1, the inhibition ratio were 68.8% and 48.0%, respectively.

Effect of baicalin on human umbilical vein endothelial cell injury induced by lipocalin-2 / 中国药学杂志

OBJECTIVE: To investigate the effect and the mechanism of baicalin in alleviating human umbilical vein endothelial cell (HUVEC) injury induced by lipocalin-2. METHODS: Lipocalin-2 at different concentration (0, 5, 10, 20 μmol·L-1) and different time gradient (0, 24, 48 and 72 h) were added in HUVEC, the proliferation and apoptosis of HUVEC, contents of interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in HUVEC supernatant, lactic dehydrogenase (LDH) activities and the expressions of p-JNK, Bax and Bcl-2 in HUVEC were measured by MTT assay, flow cytometry, enzyme-linked immuno-sorbent assay (ELISA), colorimetry and Western blot, respectively. Different concentration of baicalin (15, 30, 60 mg·L-1) or 10 μmol·L-1 SP600125, the specific inhibitor of JNK pathway, was added into HUVEC to detect the effect of baicalin. RESULTS: Compared with the control group, 10 μmol·L-1 lipocalin-2 could significantly increase the apoptosis, IL-6, MCP-1 contents and LDH activities, p-JNK expression and Bax/Bcl-2 rate but restrain the proliferation in HUVEC (P<0.05). Compared with lipocalin-2 group, 30 mg·L-1 baicalin could significantly restrain the apoptosis, IL-6, MCP-1 contents and LDH activities, p-JNK expression and Bax/Bcl-2 rate but increase the proliferation in HUVEC (P<0.05). CONCLUSION: Our findings indicate that baicalin could alleviate HUVEC injury induced by lipocalin-2, the protective mechanism is related to the inhibition of JNK pathway.

Protective effect of apoptosis signal-regulating kinase 1 inhibitor against mice liver injury

OBJECTIVE@#To explore the protective effect and its molecular mechanism of apoptosis signal-regulating kinase 1 (ASK1) inhibitor (GS-459679) on acetaminophen-induced liver injury in mice.@*METHODS@#The model of liver injury was established by administration of acetaminophen (APAP) (300 mg/kg, i.p.) on C57BL/6 mice. Forty-eight male C57BL/6 mice were randomly divided into four groups, consisting of control group, GS group (GS-459679, 30 mg/kg, i.p.), APAP-induced group, and GS combined with APAP-induced group. For GS combined with APAP-induced group, mice were treated with GS 30 min prior to administration of APAP. After mice were euthanized at 6 h or 12 h, respectively, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed, and mRNA levels of TNF-α, IL-6 and IL-1β were tested. The activity of glutathione (GSH), oxidized GSH (GSSG) and malondialdehyde were quantified. In addition, ASK1, P-ASK1, JNK and P-JNK protein levels were tested in all groups.@*RESULTS@#The ASK1 and P-ASK1 levels were up-regulated in APAP-induced group. Compared to the control group, serum levels of ALT and AST, and mRNA levels of TNF-α, IL-6 and IL-1β were increased in APAP-induced group. Meanwhile, the levels of MAD and GSSG, and the ratio of GSSG/GSH were higher and the JNK was activatedin APAP-induced group compared with that in control group. However, compared to APAP-induced group, GS combined with APAP-induced group displayed a decrease of protein expression levels of ASK1, P-ASK1 and P-JNK, a reduction of serum levels of ALT and AST, a decrease in TNF-α, IL-6 and IL-1β mRNA levels, and a low ration of GSSG/GSH.@*CONCLUSIONS@#GS-459679 treatment effectively down-regulates ASK1 and P-ASK1 expression. Addition of GS-459679 decreases the generation of liver metabolites and inflammatory factors, reduces oxidative stress reaction, inhibits JNK activation, and then protects the responsiveness to APAP-induced liver injury.

Study on Inhibitory Effects of Berberine on Vascular Endothelial Cells Injury Induced by PM2.5 and Its p38 MAPK Signal Pathway Mechanism / 中国药房

OBJECTIVE:To study the inhibitory effects of berberine on EA.hy926 human umbilical vein endothelial cells (EA. hy926 cells) injury induced by particulates with no more than 2.5 μm air aerodynamic diameter in atmospheric (PM2.5),and its p38 mitogen-activated protein kinase(MAPK)signal pathway mechanism. METHODS:PM2.5 samples were collected and hatched EA.hy926 cells with concentrations of 0(blank control),20,200 and 400 mg/L for 24 h. The survival rate and apoptosis rate of cells,contents of IL-6,TNF-α and MDA,activities of SOD and LDH,protein levels of p-p38 MAPK,Bcl-2 and Bcl-2 associated X protein (Bax) were detected. The above indexes of EA.hy926 cells in blank control group,PM2.5 group (200 mg/L PM2.5), p38 MAPK pathway-specific blocker SB203580 group (20 μmol/L SB203580+200 mg/L PM2.5),berberine low-,medium- and high-concentrations groups(5,10,20 μmol/L berberine+200 mg/L PM2.5)were also determined. RESULTS:Compared with blank control,survival rate of cells,SOD activity and Bcl protein decreased after 200,400 mg/L PM2.5 hatched;apoptosis rate of cells, contents of IL-6,TNF-α and MDA,LDH activity,protein levels of p-p38 MAPK and Bax increased (P<0.05),in concentra-tion-dependent manner. Compared with PM2.5 group,survival rate of cells,SOD activity and Bcl-2 protein increased in berberine medium-,high-concentrations groups and SB203580 group;apoptosis rate of cells,contents of IL-6,TNF-α and MDA,LDH ac-tivity,protein levels of p-p38 MAPK and Bax decreased (P<0.05). CONCLUSIONS:Berberine attenuates PM2.5-induced EA. hy926 cells injury via the inhibition of p38 MAPK pathway.

Allicin prevents EA. hy926 endothelial cell injury induced by PM2. 5 via inhibiting ERK1/2 pathway / 中国药理学通报

Aim To investigate the protective effect of allicin against EA. hy926 endothelial cell injury in-duced by PM2. 5 and the possible mechanism. Meth-ods The samples of fine particulate matter ( PM2. 5 ) were collected and made into suspension. Different concentrations of PM2. 5 ( 20 , 200 , 400 mg · L-1 ) were added to EA. hy926 cell. The viability and apop-tosis of EA. hy926 cell, the protein levels of p-ERK1/2, Bax and Bcl-2 in the EA. hy926 cell, the contents of tumor necrosis factor-α( TNF-α) , interleukin-6 ( IL-6 ) , and malonaldehyde ( MDA ) , the activities of su-peroxide dismutase ( SOD ) and lactic dehydrogenase ( LDH) in the EA. hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, enzyme-linked immunosorbent assay ( ELISA ) and colorimetry, respectively. Allicin at different con-centrations(5,20,40 mg·L-1 ) or a specific inhibitor of ERK1/2 signaling pathway PD98059 ( 20 μmol · L-1 ) was added into the EA. hy926 cell to observe the effect of allicin. Results Compared with control group, PM2. 5 significantly increased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA. hy926 cell(P<0. 05). Compared with PM2. 5 group, allicin significantly decreased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but increased the viability and SOD activity in the EA. hy926 cell ( P <0. 05 ) . Conclusion Allicin displays a significant protective effect against EA. hy926 endothelial cell injury induced by PM2 . 5 and its mechanism may be related to the attenuations of in-flammation and oxidative stress via the inhibition of ERK1/2 pathway.

Diagnostic value of radionuclide salivagram in pediatric pulmonary aspiration / 中华核医学与分子影像杂志

Objective To evaluate the diagnostic value of radionuclide salivagram in children with pulmonary aspiration.Methods From March 2012 to June 2015,a total of 62 patients (37 males,25 females;age range:2 d-14 years) with suspected pediatric aspiration pneumonia were enrolled in this retrospective study.All patients underwent gastroesophageal reflux (GER) imaging and(or) radionuclide salivagram.Detection rate of pulmonary aspiration by the two imaging techniques was compared with x2 test.Results Of 62 patients,14 were diagnosed as pulmonary aspiration,including 1 detected by GER imaging,and 13 detected by salivagram.The detection rate for pulmonary aspiration by radionuclide salivagram (26.0％,13/50) was significantly higher than that by GER imaging (3.1％,1/32;x2=7.211,P＜0.05).Eight of the 13 cases with pulmonary aspiration diagnosed by radionuclide salivagram underwent upper gastrointestinal radiography,and 5 cases had visible contrast agent in the airway.Conclusion Radionuclide salivagram has a higher detection rate for pulmonary aspiration compared to GER imaging,and has good concordance with the traditional upper gastrointestinal radiography.

Synthesis of 6-18F-Py-AMD3465 and the microPET/CT imaging of this agent in mice bearing A549 tumor / 中华核医学与分子影像杂志

Objective To synthesize 628F-Py-AMD3465,to investigate its biodistribution in mice and to perform the microPET/CT imaging on mice bearing human lung cancer cell (A549).Methods AMD3465 quaternary ammonium salt precursor was directly labeled with 18F,then 628F-Py-AMD3465 was synthesized through nucleophilic reaction,hydrolysis,neutralization and the product was purified using HPLC.The labeling yield and radiochemical purity were analyzed by HPLC.Fifteen Kunming mice were injected with 5.55 MBq of 628F-Py-AMD3465 and sacrificed at 5,20,40,60 and 120 min postinjection.The selected tissues were harvested and weighed,and the radioactivity in the tissues was measured by an automated γ-spectrometer.The ％ID/g was calculated.MicroPET/CT studies were performed on A549-bearing mice after injecting 6-18F-Py-AMD3465 through vena caudal.Paired t test was used.Results 6-18F-Py-AMD3465 was successfully synthesized with the labeling yield of (9.0±2.0)％,the total synthesis time was about 60 min,and the radiochemical purity was more than 98％.Biodistribution studies showed that the radiouptake was higher in the kidneys and bladder of normal mice,which demonstrated that 6-18 F-Py-AMD3465 was mainly excreted through the kidneys.Biodistribution in A549-bearing mice was similar to that in normal mice.The tumor/muscle ratio at 40 min was 5.0,but the radiouptake of the tumor was still lower than that of the normal lung:(8.05±0.35) ％ID/g vs (9.33±0.66) ％ID/g;t=5.26,P＜0.05.MicroPET/CT imaging showed that the high-uptake location of 6-18F-Py-AMD3465 in tumor-bearing mice was similar to the normal mice,and the tumor uptake reached the maximum level at 45 min post-injection (SUV 0.67).Conclusions 6-18F-Py-AMD3465 can be synthesized by a simple method.A lower uptake could be shown in the tumor compared to that in the lung and the tracer has limited diagnostic value for lung cancer.

Tanshinone ⅡA attenuates PM2 .5-induced vascular endothelial cell injury via p38 MAPK signal pathway / 中国病理生理杂志

AIM:To investigate the effect and the mechanism of tanshinone ⅡA in attenuating PM2.5-induced human umbilical vein endothelial EA.hy926 cell injury.METHODS:The samples of fine particulate matter (PM2.5) were collected in Guangzhou and made into suspension.Different concentrations (0, 20, 200 and 400 mg/L) of PM2.5 were added to EA.hy926 cells.The viability and apoptosis of EA.hy926 cells, the protein levels of p-p38 MAPK, Bax and Bcl-2 in the EA.hy926 cells, the contents of interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α) and malonaldehyde ( MDA) , and the activity of superoxide dismutase ( SOD) and lactic dehydrogenase ( LDH) in the EA.hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, ELISA and colorimetry, respectively.Tanshinone ⅡA at different concentrations (5, 10 and 20 μmol/L) or a specific inhibitor of p38 MAPK pathway, SB203580 (20μmol/L) , was added into the EA.hy926 cells to observe the effect of tanshinone ⅡA.RESULTS:Compared with control group, PM2.5 significantly increased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-p38 MAPK and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA.hy926 cells (P<0.05).Compared with PM2.5 group, tanshinone IIA significantly decreased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-38 MAPK and Bax/Bcl-2 ratio, but increased the viabil-ity and SOD activity in the EA.hy926 cells (P<0.05).CONCLUSION:Tanshinone ⅡA attenuates PM2.5-induced EA. hy926 cell injury via the inhibition of p38 MAPK pathway.

Protective effect of apoptosis signal-regulating kinase 1 inhibitor against mice liver injury

Objective: To explore the protective effect and its molecular mechanism of apoptosis signal-regulating kinase 1 (ASK1) inhibitor (GS-459679) on acetaminophen-induced liver injury in mice. Methods: The model of liver injury was established by administration of acetaminophen (APAP) (300 mg/kg, i.p.) on C57BL/6 mice. Forty-eight male C57BL/6 mice were randomly divided into four groups, consisting of control group, GS group (GS-459679, 30 mg/kg, i.p.), APAP-induced group, and GS combined with APAP-induced group. For GS combined with APAP-induced group, mice were treated with GS 30 min prior to administration of APAP. After mice were euthanized at 6 h or 12 h, respectively, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed, and mRNA levels of TNF-α, IL-6 and IL-1β were tested. The activity of glutathione (GSH), oxidized GSH (GSSG) and malondialdehyde were quantified. In addition, ASK1, P-ASK1, JNK and P-JNK protein levels were tested in all groups. Results: The ASK1 and P-ASK1 levels were up-regulated in APAP-induced group. Compared to the control group, serum levels of ALT and AST, and mRNA levels of TNF-α, IL-6 and IL-1β were increased in APAP-induced group. Meanwhile, the levels of MAD and GSSG, and the ratio of GSSG/GSH were higher and the JNK was activatedin APAP-induced group compared with that in control group. However, compared to APAP-induced group, GS combined with APAP-induced group displayed a decrease of protein expression levels of ASK1, P-ASK1 and P-JNK, a reduction of serum levels of ALT and AST, a decrease in TNF-α, IL-6 and IL-1β mRNA levels, and a low ration of GSSG/GSH. Conclusions: GS-459679 treatment effectively down-regulates ASK1 and P-ASK1 expression. Addition of GS-459679 decreases the generation of liver metabolites and inflammatory factors, reduces oxidative stress reaction, inhibits JNK activation, and then protects the responsiveness to APAP-induced liver injury.

Study on berberine of reducing IL-6 and TNF-α secreted in HUVEC induced by visfatin through JNK signal pathway / 中草药

Objective: To study the regulation of c-Jun N-terminal kinase (JNK) pathway on interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) secreted in human umbilical vein endothelial cell (HUVEC) induced by visfatin and the intervention of berberine. Methods: HUVEC was cultured for the in vitro experiment, visfatin was added at different concentration (0, 50, 100, and 200 μg/L) and in different times (0, 6, 12, 24, and 48 h) to obserbe the effect on HUVEC. MTT assay was used to detect the proliferation of HUVEC, apoptosis rate was measured by Flow Cytometer, the contents of IL-6 and TNF-α in HUVEC supernatant were determined by enzyme-linked immuno-sorbent assay (ELISA) assay, and the protein expressions of p-JNK, Bax, and Bcl-2 in HUVEC lysate were determined by Western blotting. Berberine (50 μmol/L) and SP600125 (10 μmol/L) were added to interfere HUVEC and to detect the effect of berberine. Results: Compared with the control group, visfatin (100 μg/L) could significantly decrease the proliferation of HUVEC, induce HUVEC apoptosis, and increase the IL-6 and TNF-α contents in HUVEC supernatant. Meanwhile, it could increase p-JNK and Bax expression, decrease Bcl-2 expression after 24 h. Compared with visfatin group, berberine and SP600125 could increase the proliferation of HUVEC while decrease the IL-6 and TNF-α contents in HUVEC supernatant, restrain p-JNK and Bax expression while increase Bcl-2 expression in HUVEC. Conclusion: Berberine could decrease the contents of IL-6 and TNF-α in HUVEC supernatant and ease the injury induced by visfatin, the protective mechanism is related to the inhibition of JNK signal pathway activation.

Effects of enhancer of zeste homolog (EZH2) downregulation on the proliferation and invasion of nasopharyngeal carcinoma cell and the possible mechanisms / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay, plate colony formation assay and flow cytometry. The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay, respectively. The expressions of EZH2 and epithelial-mesenchymal transition (EMT)-related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.</p><p><b>RESULTS</b>The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells (P < 0.001). Colony formation rate (84.44%) of 5-8F/shEZH2 cells was lower than control (31.56%, P = 0.001). Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase, with a very low ratio of cells in S phase. Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly, and the 48-hour relative migration distance of 5-8F/ShEZH2 cells and control cells was 0.58 ± 0.05, and 0.81 ± 0.02, respectively (P < 0.000). Matrigel invasion assay, showed the invasive capacity of cells silencing EZH2 was significantly inhibited, with less penetrating cells (72.23 ± 4.08) compared to control (150.95 ± 16.27), P < 0.000. The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177% and 158% respectively, and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04% and 41.18% respectively. Similar results also were obtained with Western blot analysis.</p><p><b>CONCLUSION</b>EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro, which might be mediated by inducing EMT.</p>

Flavonoids of Erigeron canadensis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the flavonoids of Erigeron canadensis.</p><p><b>METHOD</b>The constituents of EtOH extraction from the whole plant of E. canadensis were isolated and purified by repeated column chromatography. These compounds were identified by their physical and spectral data.</p><p><b>RESULT</b>Twelve flavonoids were isolated and identified as quercetin-7-O-beta-D-galactopyranoside(1),quercetin(2), luteolin(3), apigenin(4),5,7,4'-trihydroxy-3'-methoxy flavone(5), quercetin-3-alpha-rhamnopyranoside(6), quercetin-3-O-beta-D-glucopyranoside(7), apigenin-7-O-beta-D-glucopyranoside(8), luteolin-7-O-beta-D-glucuronide methyl ester(9),4'-hydroxy baicalein-7-O-beta-D-glucopyranoside(10),baicalein(11),rutin(12).</p><p><b>CONCLUSION</b>Compound 1 was isolated from the Compositae family for the first time. Compound 5 and 9 were firstly isolated from the genus Erigeron. Compound 3,4,7,8 and 11 were isolated from E. canadensis for the first time.</p>