Objective To investigate the effect of asiaticoside on blood pressure and relaxation of thoracic aorta in rats and explore the underlying mechanism.Methods SD rats treated with 50 and 100 mg/kg asiaticoside by daily gavage for 2 weeks were monitored for systolic blood pressure changes,and histological changes of the thoracic aorta were evaluated using HE staining.In isolated rat endothelium-intact and endothelium-denuded thoracic aorta rings,the effects of asiaticoside on relaxation of the aortic rings were tested at baseline and following norepinephrine(NE)-and KCl-induced constriction.The vascular relaxation effect of asiaticoside was further observed in NE-stimulated endothelium-intact rat aortic rings pretreated with L-nitroarginine methyl ester,indomethacin,zinc protoporphyrin Ⅸ,tetraethyl ammonium chloride,glibenclamide,barium chloride,Iberiotoxin,4-aminopyridine,or TASK-1-IN-1.The aortic rings were treated with KCl and NE followed by increasing concentrations of CaCl2 to investigate the effect of asiaticoside on vasoconstriction induced by external calcium influx and internal calcium release.Results Asiaticoside at 50 and 100 mg/kg significantly lowered systolic blood pressure in rats without affecting the thoracic aorta histomorphology.While not obviously affecting resting aortic rings with intact endothelium,asiaticoside at 100 mg/kg induced significant relaxation of the rings constricted by KCl and NE,but its effects differed between endothelium-intact and endothelium-denuded rings.In endothelium-intact aortic rings pretreated with indomethacin,ZnPP Ⅸ,barium chloride,glyburide,TASK-1-IN-1 and 4-aminopyridine,asiaticoside did not produce significant effect on NE-induced vasoconstriction,and tetraethylammonium,Iberiotoxin and L-nitroarginine methyl ester all inhibited the relaxation effect of asiaticoside.In KCl-and NE-treated rings,asiaticoside obviously inhibited CaCl2-induced vascular contraction.Conclusion Asiaticoside induces thoracic aorta relaxation by mediating high-conductance calcium-activated potassium channel opening,promoting nitric oxide release from endothelial cells and regulating Ca2+ influx and outflow,thereby reducing systolic blood pressure in rats.

Objective To investigate the effect of asiaticoside on blood pressure and relaxation of thoracic aorta in rats and explore the underlying mechanism.Methods SD rats treated with 50 and 100 mg/kg asiaticoside by daily gavage for 2 weeks were monitored for systolic blood pressure changes,and histological changes of the thoracic aorta were evaluated using HE staining.In isolated rat endothelium-intact and endothelium-denuded thoracic aorta rings,the effects of asiaticoside on relaxation of the aortic rings were tested at baseline and following norepinephrine(NE)-and KCl-induced constriction.The vascular relaxation effect of asiaticoside was further observed in NE-stimulated endothelium-intact rat aortic rings pretreated with L-nitroarginine methyl ester,indomethacin,zinc protoporphyrin Ⅸ,tetraethyl ammonium chloride,glibenclamide,barium chloride,Iberiotoxin,4-aminopyridine,or TASK-1-IN-1.The aortic rings were treated with KCl and NE followed by increasing concentrations of CaCl2 to investigate the effect of asiaticoside on vasoconstriction induced by external calcium influx and internal calcium release.Results Asiaticoside at 50 and 100 mg/kg significantly lowered systolic blood pressure in rats without affecting the thoracic aorta histomorphology.While not obviously affecting resting aortic rings with intact endothelium,asiaticoside at 100 mg/kg induced significant relaxation of the rings constricted by KCl and NE,but its effects differed between endothelium-intact and endothelium-denuded rings.In endothelium-intact aortic rings pretreated with indomethacin,ZnPP Ⅸ,barium chloride,glyburide,TASK-1-IN-1 and 4-aminopyridine,asiaticoside did not produce significant effect on NE-induced vasoconstriction,and tetraethylammonium,Iberiotoxin and L-nitroarginine methyl ester all inhibited the relaxation effect of asiaticoside.In KCl-and NE-treated rings,asiaticoside obviously inhibited CaCl2-induced vascular contraction.Conclusion Asiaticoside induces thoracic aorta relaxation by mediating high-conductance calcium-activated potassium channel opening,promoting nitric oxide release from endothelial cells and regulating Ca2+ influx and outflow,thereby reducing systolic blood pressure in rats.

OBJECTIVE: To demonstrate the mechanism of mechanical ventilation (MV) induced endoplasmic reticulum stress (ERS) promoting mechanical ventilation-induced pulmonary fibrosis (MVPF), and to clarify the role of angiotensin receptor 1 (AT1R) during the process. METHODS: The C57BL/6 mice were randomly divided into four groups: Sham group, MV group, AT1R-shRNA group and MV+AT1R-shRNA group, with 6 mice in each group. The MV group and MV+AT1R-shRNA group mechanically ventilated for 2 hours after endotracheal intubation to establish MVPF animal model (parameter settings: respiratory rate 70 times/minutes, tidal volume 20 mL/kg, inhated oxygen concentration 0.21). The Sham group and AT1R-shRNA group only underwent intubation after anesthesia and maintained spontaneous breathing. AT1R-shRNA group and MV+AT1R-shRNA group were airway injected with the adeno-associated virus one month before modeling to inhibit AT1R gene expression in lung tissue. The expressions of AT1R, ERS signature proteins [immunoglobulin heavy chain-binding protein (BIP), protein disulfide isomerase (PDI)], fibrosis signature proteins [collagen I (COL1A1), α-smooth muscle actin (α-SMA)] in lung tissues were detected by immunofluorescence and Western blotting. Hematoxylin-eosin (HE) staining was used to evaluate lung injury and Masson staining was used to evaluate pulmonary fibrosis. RESULTS: Compared with the Sham group, the degree of pulmonary fibrosis and lung injury were more significant in the MV group. In the MV group, the protein expressions of AT1R, BIP, PDI, COL1A1 and α-SMA were increased (AT1R/β-actin: 1.40±0.02 vs. 1, BIP/β-actin: 2.79±0.07 vs. 1, PDI/β-actin: 2.07±0.02 vs. 1, COL1A1/α-Tubulin: 2.60±0.15 vs. 1, α-SMA/α-Tubulin: 2.80±0.25 vs. 1, all P < 0.01). The number of E-cad⁺/AT1R⁺ and E-cad⁺/BIP⁺ cells in lung tissue increased, and the fluorescence intensity of COL1A1 and α-SMA increased. Compared with the MV group, the degree of pulmonary fibrosis and lung injury were significantly relieved in the MV+AT1R-shRNA group. In the MV+AT1R-shRNA group, the protein expressions of AT1R, BIP, PDI, COL1A1 and α-SMA were decreased (AT1R/β-actin: 0.53±0.03 vs. 1.40±0.02, BIP/β-actin: 1.73±0.15 vs. 2.79±0.07, PDI/β-actin: 1.04±0.07 vs. 2.07±0.02, COL1A1/α-Tubulin: 1.29±0.11 vs. 2.60±0.15, α-SMA/α-Tubulin: 1.27±0.10 vs. 2.80±0.25, all P < 0.01). The number of E-cad⁺/AT1R⁺ and E-cad⁺/BIP⁺ cells in lung tissue decreased, and the fluorescence intensity of COL1A1 and α-SMA decreased. There was no statistically significant difference in the indicators between AT1R-shRNA group and Sham group. CONCLUSIONS MV up-regulate the expression of AT1R in alveolar epithelial cells, activate the AT1R pathway, induce ERS and promote the progression of MVPF.
Mice ; Animals ; Pulmonary Fibrosis/chemically induced* ; Lung Injury ; Respiration, Artificial/adverse effects* ; Actins/metabolism* ; Tubulin ; Mice, Inbred C57BL ; Endoplasmic Reticulum Stress ; RNA, Small Interfering

ObjectiveTo investigate the changes in gut microbiota after transjugular intrahepatic portosystemic shunt (TIPS) in cirrhotic patients with mild hepatic encephalopathy (MHE) in different prognosis groups. MethodsA total of 28 MHE cirrhotic patients who were hospitalized and underwent TIPS in Xijing Hospital of Digestive Diseases from July 2016 to July 2017 were enrolled. Fecal samples and related clinical data were collected on days 1-3 before surgery and at 1 month after surgery. According to the prognosis after surgery, the patients were divided into none-hepatic encephalopathy (HE) group with 8 patients, MHE group with 12 patients, and overt hepatic encephalopathy (OHE) group with 8 patients. Fecal samples were analyzed by 16S rRNA sequencing to obtain the relative abundance of gut microbiota, and SPSS and R packages were used to analyze the biodiversity, postoperative changes, and differences in such changes of gut microbiota at the genus level between groups. The chi-square test was used for comparison of categorical data between groups; the Kruskal-Wallis H test was used for comparison of continuous data between three groups; the Bonferroni method was used for multiple comparisons of multiple samples; the Wilcoxon signed-rank test was used for comparison before and after surgery within each group. For microbiome beta-diversity analyses, a principal coordinate analysis (PCoA) was performed based on Bray-Curtis distance matrix, and the Adonis method (PerMANOVA) was used for comparison between groups. ResultsPCoA based on Bray-Curtis distance matrix showed that only the MHE group had a significant change in beta diversity after surgery (F=2.71, P=0.049). After surgery, the non-HE group had significant increases in the abundance of the native flora Dialister, Coprococcus, Ruminococcaceae_uncultured, Flavonifractor, and Clostridium_sensu_stricto_1 (Z=2.521, 2.1, 2.1, 2.1, and 1.96, all P＜0.05); the MHE group had significant reductions in the abundance of the harmful flora Granulicatella（Z=2.521,P=0.012）, Enterococcus（Z=2.51,P=0.012）, Streptococcus（Z=2.432,P=0.015）, and Rothia（Z=2.001,P=0.045） and significant increases in the abundance of Veillonella（Z=2.353,P=0.019） and Megasphaera（Z=1.955,P=0.05）; the OHE group only had a significant increase in the abundance of Veillonella after surgery (Z=2.38, P=0.017). There was a significant difference in the change in gut microbiota (postoperative abundance/preoperative abundance) between the non-HE group, the MHE group, and the OHE group ［2.00 (1.11-91.61) vs 1.21 (0.26-679) vs 0.09 (0.01-0.92), χ2=6.249, P=0.043］. ConclusionThere is a significant difference in the change in gut microbiota after TIPS between patients with different prognoses, and the increase in the abundance of native flora may have a certain influence on the remission of MHE.

OBJECTIVE:To establish a method for simultaneous determination of 5 effective components in Luohua zizhu dry extract. METHODS:UPLC-MS/MS was conducted. The separation was performed on an Zorbax Eclipse Plus C18 column with mo-bile phase of acetonitrile-water(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was set at 40℃and sam-ple size was 2 μL. The analytes were detected in the multiple reaction monitoring(MRM)mode. Nitrogen was used as drying gas and atomized gas. The temperature and flow rate of drying gas were 325 ℃ and 6 L/min. The pressure of atomized gas was 45 psi. The temperature and flow rate of sheath gas were 350℃and 12 L/min. The voltage of capillary were 4000 V(+)and 3500 V(-). The voltage of nozzle was 500 V. RESULTS:The linear ranges of luteoloside,acteoside,quercetin,luteolin and rutin were 0.5048-252.4 ng/mL(r=0.9999),0.7124-356.2 ng/mL(r=0.9990),0.5094-254.7 ng/mL(r=0.9962),0.3030-151.5 ng/mL(r=0.9998) and 0.6022-301.1 ng/mL(r=0.9996),respectively. RSDs of precision,stability and reproducibility tests were all less than 3.0%. The limit of quantitation were 0.42,0.87,0.33,0.12,0.76 ng/mL. The recoveries were 97.99%-101.20%(RSD=1.3%,n=6), 96.50%-101.20%(RSD=1.7%,n=6), 94.81%-99.34%(RSD=1.7%,n=6), 97.54%-100.51%(RSD=1.2%,n=6), 93.37%-98.70%(RSD=1.9%,n=6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible, and can be used for simultaneous determination of 5 effective components in Luohua zizhu dry extract.

Objective To investigate the expression and significance of PDCD4 and MMP-7 in human gastric carcinoma (GC) tissues and to analyze their association with clinical pathological characteristics in GC patients.Methods Immunohistochemical staining SP was used to detect the expression of PDCD4 and MMP-7 protein in gastric cancer specimens and para-carcinoma tissue in 70 cases,and the relationship with clinical pathological characteristics were analyzed.Results The expression rate of MMP-7 in GC tissues in the study group was high and significantly higher than that in the control group (78.6％ vs.22.9％,P ＜ 0.05),while the expression rate of PDCD4 protein was low and significantly lower than that in the control group (31.4％ vs.75.7％,P ＜0.05).Both the expressions of PDCD4 and MMP-7 were significantly associated with tumor lymph node metastasis,peritoneal metastasis,the depth of invasion and late clinical stage of GC (P(0.05).The correlation analysis indicated that there was a negative correlation between the expressions of PDCD4 and MMP-7 (r =-0.526,P =0.000).Conclusion The expression of PDCD4 and MMP-7 might be correlated to the carcinogenesis and invasion,and might be involved in the GC regulation.

Objective To investigate the expression and significance of PDCD4 and MMP-7 in human gastric carcinoma (GC) tissues and to analyze their association with clinical pathological characteristics in GC patients.Methods Immunohistochemical staining SP was used to detect the expression of PDCD4 and MMP-7 protein in gastric cancer specimens and para-carcinoma tissue in 70 cases,and the relationship with clinical pathological characteristics were analyzed.Results The expression rate of MMP-7 in GC tissues in the study group was high and significantly higher than that in the control group (78.6％ vs.22.9％,P ＜ 0.05),while the expression rate of PDCD4 protein was low and significantly lower than that in the control group (31.4％ vs.75.7％,P ＜0.05).Both the expressions of PDCD4 and MMP-7 were significantly associated with tumor lymph node metastasis,peritoneal metastasis,the depth of invasion and late clinical stage of GC (P(0.05).The correlation analysis indicated that there was a negative correlation between the expressions of PDCD4 and MMP-7 (r =-0.526,P =0.000).Conclusion The expression of PDCD4 and MMP-7 might be correlated to the carcinogenesis and invasion,and might be involved in the GC regulation.

OBJECTIVETo observe the effect of Rgl treatment on prognosis of alcoholic hepatitis using a rat model.

METHODSFemale Sprague-Dawley rats were radomly divided into four groups:unmodeled control, untreated model, Rgl-treated model, and dexamethasone (DXM)-treated model. The model groups were generated by intragastric injection of alcohol. The unmodeled control group was given an equal dosage of normal saline by the same route. After model establishment, the Rg1 treatment group and the DXM treatment group were administered a 120-hour treatment of Rgl or DXM; the unmodeled controls were administered normal saline on the same schedule. All rats were then fasted for 120 hours and venous blood samples were collected for detection of serum aspartate aminotransferase (AST), alanine transaminase (ALT), total bilirubin (TBil), albumin (Alb), tumor necrosis factor-alpha (TNFat) and interleukin 6 (IL-6). Markers of liver inflammation were measured by immunohistochemistry, western blotting, and real-time quantitative reverse transcription PCR. Fat and apoptosis indices were assessed by hematoxylin-eosin staining and TUNEL assay, respectively. The t-test and F test were used for statistical analyses.

RESULTSThe model group showed remarkably more liver steatosis (over one-third of the tissue) than the unmodeled control group, indicating proper establishment of alcoholic liver disease in the modeled rats. The AST, ALT, TBil, and IL-6 levels were significantly higher in the untreated model group than in the Rgl-treated group and the DXM-treated group. The values were significantly different between the Rg1-treated group and the DXM-treated group:ALT, 69.19+/-8.00 U/L vs.102.88+/-5.16 U/L; TBil, 0.36+/-0.07 µmol/L vs.1.20+/-0.18 µmol/L; IL-6, 126.50+/-6.50 U/ml vs.169.19+/-7.68 U/ml; TNFa, 268.31+/-13.19 µg/L vs.318.94+/-7.87 µg/L (all P less than 0.05). Expression of caspase3 and caspase8 was significantly higher in the model group than in the Rgltreated group and the DXM-treated group (both P<0.05). The apoptosis index was significantly lower in the Rgltreated group and the DXM-treated group than in the model group (both P<0.05). The mRNA and protein expression of caspase3, caspase8 and NF-kB were significantly lower in the Rgl-treated group and the DXM-treated group than in the model group (allP less than 0.05), and the levels of all were significantly lower in the Rgl-treated group cornered to the DXM-treated group (all P<0.05). Conehision In rats with alcoholic hepatitis, Rg1 can significantly relieve pathological injury, improve liver function by blocking the apoptotic pathway, and inhibit release of inflammatory cytokines.

Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Bilirubin ; Cytokines ; Disease Models, Animal ; Ethanol ; Female ; Ginsenosides ; Hepatitis, Alcoholic ; NF-kappa B ; Rats ; Rats, Sprague-Dawley

Objective To study the relationships between tissue damage and the ability of the pancreatic cells to regenerate ,and analyze the alteration of the pancreatic cells regeneration .Methods Sixty rats were divided into two groups :impact group(the pan‐creas was injured by a BIM‐Ⅲ biotical impact machine ,40 rats) and control group(sham operated ,20 rats) .All rats were sacrificed at 6 h ,24 h ,72 h ,7 d after operation .The level of AMS ,LPS in the serum were detected by spectrophotometry ,pancreatic cells re‐generation were examined and analyzed by TUNEL staining and flow cytomertry ,and the Bcl‐2 and Bax expression were measured by Western blot .Results In the impact groups ,LPS was activated later than AMS ,and lasted persistently .The results from TUNEL stain ,flow cytometry and Western blot indicated that pancreatic trauma induces cell death and the compensatory prolifera‐tion of pancreatic cells .The characteristics of pancreatic cells regeneration in the animal model of isolated pancreatic trauma indicate that the proper remedial time is in the first 24h after the pancreatic trauma .Conclusion Detecting AMS and LPS at the same time can help us to determine the exocrine function of pancrease .

Carbon dots(CDs)were firstly prepared via a simple pyrolysis route using citric acid as the carbon precursor and glycine as modifier. The obtained CDs were further purified by n-butyl alcohol, which made the particle size more uniform and the fluorescence intensity stronger. When excited at 380 nm, the maximum emission wavelength of the CDs was about 480 nm, with a quantum yield of 47%. Based on fluorescence quenching, a novel method for chloramphenicol assay was developed with glycine-passivated CDs. This method is simple and rapid. A linear relationship between the change of fluorescence intensity and the concentration of chloramphenicol was obtained with a relation coefficient of 0. 999 7. The recovery was in the range of 99% to 101% with a relative standard deviation of 0. 3%, which shows CDs′ potential application in drug detection.