The investigation on the leaves of Morus alba L. was carried out to find the relationship of the constituents and the pharmacological activities. The isolation and purification were performed by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography and so on. Further detailed investigation on the fraction of the ethanol extract of leaves of Morus alba yielded four Diels-Alder type adducts mulberrofuran F1 (1), mulberrofuran F (2), chalcomoracin (3), kuwanon J (4), together with two chalcones morachalcone A (5), isobavachalcone (6), and three flavones norartocarpetin (7), kuwanon C (8), 6-geranylapigenin (9). Their structures were elucidated by the spectral analysis such as NMR, MS etc. Compounds 1, 6 were isolated from this plant for the first time, compounds 4-5, 7-9 were isolated from the leaves of Morus alba L. for the first time, among which 1 was a new compound. Compounds 1-5 were evaluated for the cytotoxicity against A549, Be17402, BGC823, HCT-8 and A2780 cell lines in vitro by MTT method, but only compounds 1-3 showed cytotoxicity against several human cancer cell lines.

The investigation on the chemical constituents of Anabasis aphylla L. was carried out by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Further detailed investigation on the fraction of the ethanol extract of Anabasis aphylla L. yielded one new compound p-acetyl-phenol 1-O-beta-D-xylopyranosyl-(1-->2)-beta-D-glucopyranoside (1), together with five known compounds: piceine (2), isorhamnetin (3), quercetin (4), rutin (5) and isorhamnetin-3-rutinoside (6). Their structures were elucidated by spectral analysis such as NMR and MS. Among these compounds, compounds 2-6 were isolated from this plant for the first time.

Objective To investigate the possible mechanism that hepatocyte growth factor (HGF) inhibits renal tubular epithelial-mesenchymal transition (EMT),and to determine whether Smurf2 expression induced by TGF-β1 can be reversed by HGF in normal rat kidney epithelial cells (NRK-52E).Methods Using rat NRK-52E cell line as an in vitro system,NRK-52E cells were incubated with 5 μg/L TGF-β1 for 0-24 h.Part of cells were pretreated with 20 μg/L HGF for 30 min or not,then incubated with or without 5 μg/L TGF-β1 for 1 h or 48 h.The other cells were transfected with pFlag-Smurf2 or Smurf2 siRNA for 24 h,then treated with or without 20 μg/L HGF for 24 h.The expressions of Smurf2,SnoN,E-cadherin,alpha-smooth muscle actin (α-SMA) and fibronectin (FN) were detected by Western blotting and indirect immunofluorescence staining assays.Results Compared to normal control,TGF-β1 could rapidly induce Smurf2 protein expression in a short time (P＜0.01).Meanwhile,the expressions of FN and α-SMA were significantly induced,and the expression of E-cadherin was reduced in NRK-52E cells by TGF-β1.In contrast,in the NRK-52E cells pretreated with HGF,HGF could obviously inhibit Smurf2 expression induced by TGF-β1,and reversed the down-regulation of SnoN (P＜0.01) and E-cadherin (P＜0.05),the up-regulation of α-SMA (P＜0.01) and FN (P＜0.01) induced by TGF-β1.Moreover,overexpression of Smurf2 in NRK-52E cells could partly inhibit the up-regulation of SnoN protein by HGF,while down-regulation of Smurf2 could up-regulate the expression of SnoN induced by HGF.Conclusions HGF can abolish EMT induced by TGF-β1 in renal tubular epithelial cells through down-regulating Smurf2 expression and suppressing ubiquitin-proteasome dependent degradation of SnoN.

Ten flavonoids were isolated from the 95% ethanol extract of the seeds of Alpinia galanga Willd. with a combination of various chromatographic techniques, including silica gel, Sephadex LH-20 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (2R, 3S)-pinobaksin-3-cinnamate (1), (2R, 3R)-pinobaksin-3-cinnamate (2), pinocembrin (3), pinobaksin (4), 3-O-acetylpinobaksin (5), galangin (6), galangin-3-methylether (7), kumatakenin (8), 3-methylkaempferol (9) and (2R, 3R)-3, 5-dihydroxy-7-methoxyflavanone (10). Among them, compound 1 is a new compound, compounds 2, 5 and 10 were isolated from the genus Alpinia for the first time, and others were isolated from this plant for the first time.

Objective To investigate the relationship between high-glucose-induced fibronectin(FN) expression and up-regulation of integrin-linked kinase(ILK) in human kidney tubular epithelial cells (HKC) and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin (STZ)-indueed diabetic model of CD-1 mice were enrolled in this study.Western blot was used to detect the expression of FN and ILK.The kinase dead ILK plasmid (pCMV-kdlLK) were transferred to HKC. Results Four weeks after injection of STZ,CD-1 mice had higher blood glucose level as compared to the control [(20.3±2.7) mmol/L vs (6.1±1.4) mmol/L,P＜0.01].Meanwhile,expression of FN and ILK was significantly increased in diabetic mice as compared to the control (P＜0.01).There was positive correlation between the expression of FN and ILK (r=0.899,P＜0.01).High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner.Blockage of ILK activation by pCMV-kdILK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression.Blockage of ILK activation abrogates this effect.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To investigate the effect of high glucose on renal tubular epithelial-mesenchymal transition,and to analyze the relationship between high glucose and transforming growth factor β1(TGF-β1)and the mechanism of renal interstitial fibrosis. Methods HKC and Smad7-overexpression HKC cells were grown in DMEM/F12 medium containing 5%～10%newborn calf serum.They were cultured for 16 h in free serum medium after 80%cells were adhered onto the surface of the flask.Afterwards,they were stimulated by high glucose(glucose concentration:25 mmol/L and 50 mmol/L).The expression of α-SMA,E-cadherin and fibronectin was detected by Western blot while the supernatant level of TGF-β1 was detected by ELISA.Cell motility and migration was evaluated using Boyden chamber motogenicity assay. Results In HKC induced by high glucose,the expression of α-SMA and fibronectin protein was highly upregulated while the expression of E-cadhefin protein was down-regulated.The expression of TGF-β1was up-regulated in a dose-dependent manner.These above-mentioned effects could be obviously inhibited by anti-TGF-β1 antibody.The protein expression of α-SMA,fibronectin and E-cadherin had no obvious change in Smad7-overexpression HKC induced by high glucose.HKC exhibited enhanced motility and invasive capacity in high glucose groups,compared to that in control group.Migrated cell counting was(12.4±3.7)and(18.6±4.4)cell/HP in 25 and 50 mmol/L glucose groups respectively. Conclusion High glucose may induce renal tubular epithelialmesenchymal transition through TGF-β1 pathway,which can be inhibited by blocking the Smad signal pathway.

Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).

Objective To study the pathological changes and expressions of NO and iNOS mRNA in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the lung injury.Methods A total of 140 male SD rats were randomly (random number) ivided into the room temperature (25 ℃) environment traumatic hemorrhagic shock group (room temperature group) and the dry heat traumatic hemorrhagic shock groups (dry heat group,temperature 40℃,humidity 10％),respectively,and each groups was further randomly divided into 7 subgroups:the control subgroup,post shock subgroups at 0,0.5,1,1.5,2and 3 h (n =10 in each subgroup).The rats of control subgroup were not treated,and rats of dry heat group were placed in dry heat environment for 60 min,then anesthetized,fixed,and insertion of intravenous indwelling needles and catherization of right carotid artery,jugular vein and the right femoral artery were performed.After stabilization for 10 min,2500 g iron wheel was used to be dropped from 30 m height and vertically hit the upper left femoral of SD rats in order to make comminuted fracture,wounds were quickly dressed after injury.Exsanguination from right femoral artery was kept until MAP maintained at (35 ± 5) mmHg,and resuscitation was carried out after continue monitoring for 60 min.After the establishment of traumatic hemorrhagic shock model in each environment,the rats were sacrificed at given intervals,and thoracotomy was performed to take broncho-alveolar lavage fluid (BALF) and lung tissue.Pathological changes of lung tissues were observed by using HE staining and NO concentration of lung tissue was detected by one-step method,and changes of the iNOS mRNA expressions were detected by using fluorescence quantitative PCR.Then t test,ANOVA and Pearson correlation analysis were used for the data analysis.Results The pathological change in dry heat group at each interval was more severe,and pulmonary histopathological injury score was higher,and the protein exudation was more profuse compared with the room temperature group.NO concentration in lung tissue homogenate of dry heat group was higher than that of room temperature group (t =2.472,P ＜ 0.05),and the difference in NO level between different intervals within the dry heat group was statistically significant (F =6.77,P ＜ 0.01).The NO concentration in dry heat group reached its maximum at 2 h (3.35 ± 0.23) μmol / g and the peak value emerged sooner than that in room temperature group.The difference was statistically significant in overall expression of iNOS mRNA between two groups analyzed with t test (t =3.619,P ＜ 0.01),and there was statistically significant difference between intervals within the dry heat group (F =12.34,P ＜0.01).The values of iNOS mRNA in the dry heat group were higher than those in the room temperature group at the same given intervals,and the peak value appears at 1.5 h in dry heat group,and the room temperature group it began to increase at 2 h.The concentration of NO and the expression of iNOS mRNA were positively correlated with each other in two groups (r =0.680,r =0.376).The expression of iNOS mRNA and lung histopathological injury score was positively correlated in two groups (r =0.846,r =0.899).Conclusions When traumatic hemorrhagic shock occurred in the dry heat desert environment,the lung injury was more severe and appeared sooner than that in the room temperature environment.NO and iNOS played important roles in the secondary lung injury in the wake of traumatic hemorrhagic shock in rats under the dry heat environmengt of desert.

The investigation on Salvia przewalskii Maxim was carried out to find the relationship of the constituents and their pharmacological activities. The isolation and purification were performed by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography, etc. Further investigation on the fraction of the 95% ethanol extract of Salvia przewalskii Maxim yielded przewalskin Y-1 (1), anhydride of tanshinone-II A (2), sugiol (3), epicryptoacetalide (4), cryptoacetalide (5), arucadiol (6), 1-dehydromiltirone (7), miltirone (8), cryptotanshinone (9), tanshinone II A (10) and isotanshinone-I (11). Their structures were elucidated by the spectral analysis such as NMR (Nuclear Magnetic Resonance) and MS (Mass Spectrometry). Compound 1 is a new compound. Compounds 4 and 5 are mirror isomers (1 : 3). Compounds 4, 5, 6, 8, 11 were isolated from Salvia przewalskii Maxim for the first time.