Isolation and identification of antitumor constituents from Trichosanthes tricuspidata / 中国药科大学学报

@#Ten compounds were isolated and purified from the dichloromethane fraction of Trichosanthes tricuspidata roots by silica gel and ODS column chromatographies. Their chemical structures were identified on the basis of their physical and chemical properties as well as the spectral data. These isolated compounds were elucidated as khekadaengoside O(1), a new hexanorcucurbitane glycoside, together with nine known compounds, including khekadaengoside C-E, K(2-5), cucurbitacin J-2-O-β-glucopyranoside(6), cucurbitacin K-2-O-β-glucopyranoside(7), cucurbitacin B, J, K(8-10). In addition, the anti-tumor activity of compounds 1-10 were evaluated in hepatocellular carcinoma BEL-7402 cell line. Among them, compounds 8-10 showed potent antitumor activity.

Inhibitory effect of jianpi-jiedu prescription-contained serum on colorectal cancer SW48 cell proliferation by mTOR-P53-P21 signalling pathway / 中南大学学报(医学版)

To investigate the effect of jianpi-jiedu (JPJD) prescription-contained serum on colorectal cancer SW48 cell proliferation and the underlying mechanisms.
 Methods: Crude extract from JPJD was made by water extract method and the main components of crude extract from JPJD were analyzed by ultra-performance liquid phase high resolution time of flight mass spectrometry (UPLC-Q-TOF/MS). The low, medium, and high-concentration of JPJD-contained serum were prepared by the serum pharmacological method. The effect of serum containing JPJD on SW48 cell proliferation was determined by MTT assay. The cell cycle was detected by flow cytometric method. The protein levels of mammalian target of rapamycin (mTOR), phospho-mTOR, P-P53, and -P21, and the mRNA level of mTOR were examined by Western blot and RT-PCR, respectively.
 Results: Seven compounds including calycosin-7-glucoside, astragaloside, ginsenoside-Re, ginsenoside-Rb1, glycyrrhizinic acid, apigenin, atractylenolide-II were identified. MTT assays demonstrated that the SW48 cell proliferation was inhibited by medium and high concentration of JPJD-contained serum and the percentages of cells at G1 phase in SW48 cell cultured in the medium and high concentration of JPJD serum group were significantly higher than those in the control group (P<0.05). Meanwhile, the levels of mTOR mRNA and phospho-mTOR protein in the medium and high concentration of JPJD serum groups were substantially lower than those in the control group (P<0.05). Conversely, the expressions of phospho-P53 and P21 protein were significantly increased in the medium and high concentration of JPJD serum group compared with those in the control group.
 Conclusion: JPJD prescription-contained serum can inhibit SW48 cell proliferation, which may be related to mTOR-P53-P21 signaling pathways.

Effect of jianpi-jiedu formula on tumor angiogenesis-relevant genes expression in colorectal cancer / 中南大学学报(医学版)

To investigate the effect of the jianpi-jiedu formula (JPJD) on the expression of angiogenesis-relevant genes in colon cancer.
 Methods: Crude extract was obtained from JPJD by water extract method. The effect of JPJD crude extract on colon cancer cell proliferation capacity was determined by MTT assays. The IC50 value was calculated by GraphPad Prism5 software. Affymetrix gene expression profiling chip was used to detect significant differences in expressions of genes after JPJD intervention, and pathway enrichment analysis was performed to analyze the differentially expressed genes. Ingenuity Pathway Analysis software was applied to analyze differentially expressed genes relevant to tumor angiogenesis based on mammalian target of rapamycin (mTOR) signaling pathway and then the network diagram was built. Western blot was used to verify the protein levels of key genes related to tumor angiogenesis.
 Results: JPJD crud extract inhibited the proliferation capacity in colon cancer cells. The IC50 values in 24, 48, and 72 hours after treatment were 13.060, 9.646 and 8.448 mg/mL, respectively. The results of chip showed that 218 genes significantly upgraded, and 252 genes significantly downgraded after JPJD treatment. Most of the genes were related to the function of biosynthesis, metabolism, cell apoptosis, antigen extraction, angiogenesis and so on. There were 12 differentially expressed angiogenesis genes. IPA software analysis showed that the JPJD downregulated expression of sphingomyelin phosphodiesterase 3 (SMPD3), VEGF, vascular endothelial growth factor A (VEGFA), integrin subunit alpha 1 (ITGA1), cathepsin B (CTSB), and cathepsin S (CTSS) genes, while upregulated expressions of GAB2 and plasminogen activator, urokinase receptor (PLAUR) genes in the colorectal cancer cell. Western blot results demonstrated that JPJD obviously downregulated expressions of phospho-mTOR (P-mTOR), signal transducer and activator of transcription 3 (STAT3), hypoxia inducible factor-1α (HIF-1α), and VEGF proteins, while obviously upregulated the level of phospho-P53 (P-P53) protein.
 Conclusion: JPJD may inhibit colorectal tumor angiogenesis through regulation of the mTOR-HIF-1α-VEGF signal pathway.

Isolation and identification of Withaphysalins from Physalis minima / 中国药科大学学报

@#To investigate the chemical constituents of Physalis minima L. , compounds were isolated by chromatographic methods from Physalis minima L. . Their structures were determined on the basis of spectral analysis. Five compounds were isolated and identified as Withaphysalin P(I), 14, 18-di-O-acetylwithaphysalin C(II), Withaphysalin Q(III), Withaphysalin 1(IV)and Withaphysalin 2(V). Compounds IV and V are new compounds, orderly named as Withaphysalin T and Withaphysalin U.

Anatomy of the superficial epigastric artery perforator flap in rats / 解剖学报

Objective To investigate the anatomy of the superficial epigastric artery perforator flap , and to provide anatomical basis for harvesting flap .Methods Of 27 SD rats, 7 were used for gross anatomy observation and anatomic characteristics and 20 rats for lead oxide-gelatin injection followed by computer picture processing , measurements and the related parameters recording .Results The superficial epigastric artery originated from femoral artery , and gave off its first branch when passed through the superficial fascia .The trunk branched into a lateral perforator and a medial perforator , which anastomosed with thoracodorsal artery and lateral thoracic artery , respectively .The average external diameter of superficial epigastric artery was (0.46 ±0.02)mm at its starting point,and(0.46 ±0.02)mm at the superficial fascia level . The nutritive area of superficial epigastric artery was (18.37 ±3.67) cm2 .The anastomosed area with thoracodorsal artery and lateral thoracic artery was(5.34 ±0.86)cm and(6.28 ±0.29)cm, respectively, away from the horizontal line through axillary,and (4.38 ±0.38)cm and(2.04 ±0.33)cm, respectively, away from the ventral median line.Conclusion The position and external diameter of superficial epigastric artery are constant , and the superficial epigastric artery perforator flap is a ideal flap model for research on free flap transplantation , flap supercharging , and hemodynamics .

Effect of rapamycin on the proliferation and apoptosis of glomerular mesangial cells / 中华肾脏病杂志

Objective To investigate the effect of different concentrations of rapamycin on the proliferation and apoptosis of glomerular mesangial cells(GMCs)and to investigate the mechanism. Methods GMCs were treated with different concentrations of rapamycin(1 μg/L,2 μg/L,4 μg/L,8 μg/L,16 μg/L).After treatment for 24 h,48 h and 72 h,cell proliferation was assessed bv MTT colorimetric assay and the growth curve was traced.After treatment for 72 h,the cell cycle distribution and the apoptotic rate of GMCs in different concentrations of rapamycin were analyzed bv flow cytometry.The effects of different concentrations of rapamycin on the mRNA and protein expression of p27 and p53 were detected by RT-PCR and Western blot respectivelyResult The low dose of rapamycin(1 μ/L)could signiticanfly inhibit the proliferation of GMCs and showed no effect on apoptosis.The high dose of rapamycin (8-16 μg/L)could significantly increase the apoptotic rate of GMCs.Rapamycin could increase the mRNA and protein expression of p27 and p53. Conclusion Rapamycin can inhibit GMCs proliferation and promote GMCs apoptosis by increasing the expression of p27 and p53.

Study on the changes of mammalian target of rapamycin with aging in rat kidneys / 中华老年医学杂志

ObjectiveTo investigate the changes in mammalian target of rapamyein (mTOR) with aging in rat kidneys.MethodsMale Wistar rats at the ages of 3, 12, 24 months were used for this study. Therenaltissuesandmesangialcellswereprocessedfor senescenceassociated β-galactosidase (SA-β-gal) staining. The expression and location of roTOR in kidneys and mesangial cells were studied by immunohistochemistry and immunocytochemistry, respectively. The mRNA and protein levels of the roTOR and p-roTOR were detected by Western blot assay and RT-PCR,respectively.ResultsThe expression of neutral β-galactosidase activity was increased in kidneys and mesangial cells with advancing age. Percentages of SA-β-gal staining positive ceils were (11.9±3.6)% versus ( 39.0±4.0)% versus ( 86.9±7.4) % in young, middle and aging glomerular mesangial cells (P<0.05). The mTOR staining appeared in the mesangial matrix and interstitium in kidneys, while the mTOR protein showed localization in cytoplasm and nucleus in mesangial cells. The staining intensity of mTOR in kidneys and mesangial cells in aged rats was markedly increased as compared to that in young and middle aged rats (P<0.05). The mRNA level of roTOR was significantly increased in kidneys and mesangial cells of agedrats versus young and middle aged rats,meanwhile, the roTOR and p-mTOR protein expressions were dramatically increased with advancing age (P<0.05 ).ConclusionsmTOR expression is increased with aging, which may play an important role in the aging process of kidneys.

Clinical Study on the Method of Reinforcing Qi,Strengthing Spleen and Clearing Heat Antitoxicant in Treating Ulcerative Colitis / 中国中医药信息杂志

Objective To investigate the therapeutic effect on ulcerative colitis by methods of reinforcing qi,strengthing spleen and clearing heat antitoxicant. Methods 99 patients were divided into two groups randomly:the treatment group for oral and enteroclyste use with TCM following in the principle mentioned above,the control group for oral and enteroclyste use with western medicine following in the common practice. Comparative studies on the total therapeutic effect and the improvement of TCM syndromes between the two groups were made. Meanwhile,the Pearson correlation analysis were made to explore the relationship between the clinical effectiveness and impact factors in two groups. Results There were significant differences in the total curative effect and the improvement of TCM syndromes between two groups (P