Nitrogen metabolism and secondary metabolism regulation of Atropa belladonna by exogenous NO under NaCl stress / 中国中药杂志

Atropa belladonna seedlings were used as experimental materials and cultivated by soil culture method. Different concentrations(0,0.05,0.1,0.2,0.5 mmol·L~(-1))of NO donor sodium nitroprusside(SNP) were sprayed on the leaves. The effects of different concentrations of SNP and different treatment time(4,8,12,16 d) on nitrogen metabolism, secondary metabolite content, precursor content of tropane alkaloid synthesis pathway and expression of key enzyme genes under 100 mmol·L~(-1) NaCl stress were studied. The results showed that with the prolongation of salt stress, the nitrogen metabolism and the accumulation of secondary metabolites of A. belladonna were inhibited to some extent. After treatment with different concentrations of exogenous SNP, the ammonium nitrogen content decreased dramatically, and the contents of nitrate nitrogen, free amino acid, soluble protein and the activities of key enzymes of nitrogen metabolism(NR, GS, GDH) were all greatly improved; the contents of precursor amino acids(ornithine, arginine) and polyamines(Put, Spd, Spm) in the secondary metabolic pathway have increased to varying degrees. The qRT-PCR analysis showed that exogenous SNP treatment can effectively promote the high expression of key enzyme genes PMT, TRⅠ and H6H in the secondary metabolic pathway of A. belladonna, and the production of hyoscyamine and scopolamine were increased notably. In summary, the application of appropriate concentration of SNP can effectively alleviate the inhibition of salt stress on the nitrogen metabolism and secondary metabolism of Atropa belladonna, and enhance its salt tolerance. Overall, 0.1 mmol·L~(-1) and 0.2 mmol·L~(-1) SNP treatment achieved the most remarkable effect.

Antioxidant and Antiapoptotic Polyphenols from Green Tea Extract Ameliorate CCl-Induced Acute Liver Injury in Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the phenolic composition, antioxidant properties, and hepatoprotective mechanisms of polyphenols from green tea extract (GTP) in carbon tetrachloride (CCl)-induced acute liver injury mouse model.@*METHODS@#High-performance liquid chromatography was used to analyze the chemical composition of the extract. Antioxidant activity of GTP was assessed by O, OH, DPPH, and ferric-reducing antioxidant power (FRAP) assay in vitro. Sixty Kunming mice were divided into 6 groups including control, model, low-, medium-, and high-doses GTP (200, 400, 800 mg/kg) and vitamin E (250 mg/kg) groups, 10 in each group. GTP and vitamin E were administered at a level of abovementioned doses twice per day for 7 days prior to exposure to a single injection of CCl. Hepatoprotective effects of GTP were evaluated in a CCl-induced mouse model of acute liver injury, using commercial enzyme linked immunosorbent assay kits, histopathological observation, terminal deoxynucleotidyl transferase-mediated dUTPNick-end labeling (TUNEL) assay and Western blot.@*RESULTS@#GTP contained 98.56 µg gallic acid equivalents per milligram extract total polyphenols, including epicatechingallate, epigallocatechin gallate, epicatechin, and epigallocatechin. Compared with the model group, low-, medium-, or high doses GTP significantly decreased serum levels of alanine aminotransferase and aspartate transaminase (P<0.01). Histopathological observation confirmed that pretreatment of GTP prevented swelling and necrosis in CCl-exposed hepatocytes. Hepatoprotective effects of low-, medium-, and high-dose GTP were associated with eliminating free radicals and improving superoxide dismutase, catalase, and glutathione peroxidase activity in the liver. Additionally, low-, medium-, and high-dose GTP decreased cell apoptosis in the CCl-exposed liver (P<0.01). Phosphorylated nuclear factor kappa-B (NF-κB), p53, Bcl-2 associated x protein/B-cell lymphoma/leukemia-2 gene, cytochrome C, and cleaved caspase-3 levels were downregulated compared with the model group (P<0.01).@*CONCLUSION@#GTP achieves hepatoprotective effects by improving hepatic antioxidant status and preventing cell apoptosis through caspase-3-dependent signaling pathways.

Cloning and expression analysis of calmodulin gene from Pinellia ternate / 中国中药杂志

According to the data of Pinellia ternate transcriptome,two calmodulin genes were cloned and named as Pt Ca M1 and PtCa M2 respectively. The results of bioinformatics analysis showed that Pt Ca Ms genes contained a 450 bp open reading frame,encoding149 amino acids.The identity of the coding sequences was 80%,and the identity of amino acids sequence was 91%. Pt Ca Ms genes contained EF-hand structure domain,belonging to the Ca M families. The Real-time PCR analysed the expression patterns of Pt Ca Ms in different tissues and different treatments. RESULTS:: showed that Pt Ca M1 and Pt Ca M2 gene were the highest expression level in tuber. Under Ca Cl2 treatment,the expressions of Pt Ca Ms were significantly higher than the control. Under EGTA,La Cl3 and TFP treatments,the expression level of Pt Ca Ms decreased gradually. In this study,the Pt Ca Ms gene were successfully cloned from P. ternate,which laid a foundation for the functional characteristic of Pt Ca Ms gene and the synthesis of alkaloids from P. ternata for further study.

Dynamic Changes of IL-17, occludin, and ZO-1 in Mice with Post-infectious Irritable Bowel Syndrome / 中山大学学报(医学科学版)

[Objective] To observe the dynamic changes of intestinal IL-17,occludin,and ZO-1 in mice with postinfectious irritable bowel syndrome (PI-IBS).[Methods] Forty C57BL/6 mice were randomly divided into 5 groups:control group and infection groups (2 weeks,4 weeks,6 weeks,and 8 weeks after trichinella infection).Infection groups were given by gavaging of 400～500 Trichinella spiralis in 0.2 mL of normal saline.The body weight of mice were recorded at week 2,4,6,and 8 after infection.The visceral sensitivity of mice was measured by the abdominal withdrawal reflex (AWR).The stool was collected continuously for 8 hours to calculate the percentage of fecal water content.Pathological changes of gut were observed by HE staining.The expressions of IL-17,occludin,and ZO-1 in ileocecus and colon were detected by immunohistochemistry and Western blotting.[Results] At week 2 after infection,the acute inflammation of the intestinal tract was observed and the body weight of mice were significantly decreased (P=0.000).Until week 8 after infection,the intestinal inflammation and body weight of mice recovered to normal.When the colorectal dilatation capacity was 0.35 and 0.5 mL,the AWR scores in the infection groups were significantly higher than those in the control group (P＜0.01).The percentages of fecal water content in the infection groups were significantly higher than those in the control group (P＜0.05).Compared with the control group,the expressions of IL-17 were significantly decreased in week 2 group (P＜0.05) and increased in week 8 group (P＜0.05).The expressions of occludin and ZO-1 in the infection groups were significantly lower than those in the control group (P＜0.05).[Conclusion] The dynamic changes of IL-17 and the decrease of Tight junction proteins may be one of the mechanisms of visceral hypersensitivity and increased percentages of fecal water content.They may be involved in the development of PI-IBS.

Notch signaling in bone marrow mesenchymal stem cells induced by li-popolysaccharide / 中国病理生理杂志

AIM:To investigate the roles of Notch signaling in lipopolysaccharide(LPS)-induced proliferation and secretion of interleukin-6(IL-6)and chemokine CXCL1 in bone marrow mesenchymal stem cells(BMSCs). METHODS:BMSCs were isolated by whole bone marrow culture.The expression levels of Notch signaling pathway recep-tors and ligands in the BMSCs treated with LPS were measured by qPCR and Western blot.The proliferation of BMSCs was analyzed by MTT assay and viable cell counting.The secretion levels of IL-6 and CXCL1 induced by LPS were measured by ELISA.RESULTS:Treatment with LPS at 1 mg/L effectively induced the proliferation of BMSCs and the secretion of IL-6.Obvious expression of Notch receptors and ligands in the BMSCs was observed,and LPS had little effect on the mRNA and protein levels of Notch receptors and ligands,but LPS increased the protein levels of Hes1 and Hey1,the target genes of Notch signaling.LPS at 1 mg/L increased the proliferation of BMSCs,whereas DAPT(Notch signal inhibitor)reduced the basal and LPS-induced proliferation of BMSCs(P<0.01).LPS treatment robustly increased the secretion of IL-6 and CXCL1 as assessed by ELISA.However,inhibition of Notch signaling almost completely abolished LPS-induced secretion of IL-6 and CXCL1(P <0.05).CONCLUSION: Inhibition of Notch signaling reduced not only the proliferation of BMSCs but also IL-6 and CXCL1 secretion induced by LPS.

Protective effect of Sanhuangyinchi Fang drug serum on hydrogen peroxide-induced DNA oxidative damage in LO2 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen peroxide-mediated DNA oxidative damage in LO2 cells.</p><p><b>METHODS</b>The LO2 cells were randomly divided into the control group, H(2)O(2) group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively.</p><p><b>RESULTS</b>Compared with H(2)O(2) group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT, GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced tailing ratio, tail length, tail moment and Olive tail moment (P<0.05).</p><p><b>CONCLUSION</b>SF drug serum, especially at the concentration of 15%, can protect LO2 cells from H(2)O(2)-mediated DNA oxidative damage.</p>

Diagnostic value of ROP with the changes of serum IGF-I, VEGF levels in premature / 中国基层医药

Objective To observe the changes of serum IGF-I,VEGF levels in premature infants,and to dis-cuss the predicting values for the development of ROP;Methods Preterm infants admitted into those whose birth weight ( BW)≤1 500g and ( or) gestational age ( GA)≤31 weeks joined in ROP-screen,and those who were diag-nosed as ROP according to ICROP were included into ROP group.In the same time,those who didn't develop ROP were divided into the control group.Serum IGF-I,VEGF were measured one week from their birth or correct gestational age of 31 weeks.Serum IGF-I, VEGF were evaluated with ELISA method.Results The concentration of IGF-I, VEGF:Group ROP[(20.65 ±11.11)μg/L,(195.61 ±105.98)ng/L] were all significantly lower than the control group [(42.86 ±18.72)μg/L,(363.59 ±101.08)ng/L](all P<0.01).There were significant different of birth weight[ROP group (1 229.69 ±232.37)g,the control group (1 401.32 ±171.78)g;t=4.144)],GM[ROP group (3.20 ±1.23)mmol/L,the control group (3.70 ±1.17)mmol/L;t=2.082],the time of oxygen inhaling[ROP group (32 ±22)d,the control group (21 ±12)d;t=-2.856],oxygen inhalation mode(the percentage of mechanical ventilation)[ROP group 33/48(68.8%),the control group 16/50(32.0%);χ2 =16.363] and fertilization way(the percentage of test tube baby)[ROP group 13/48(27.1%),the control group 4/50(8.0%);χ2 =6.262] between ROP and control group (all P<0.05).Conclusion The low serum IGF-I(<28.08μg/L),VEGF(<232.89ng/L) of a premature in the first week or correct gestational age of 31 weeks and (or)BW≤1 500g would probably suggest the development of ROP.