Azathioprine(AZA) and 6-mercaptopurine(6-MP) have a well-established effect as an immunosuppressive agent for inflammatory bowel disease(IBD),Myelosuppression is the most serious side effect during treatment.Thiopurine methyltransferase(TPMT) plays an important role in the metabolism of 6-MP. This article reviews the literature on the role of TPMT measurement prior to treatment of IBD with AZA/6-MP.

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.

Objective:To explore stigma and related factors among family members of patients with mental disorders in psychiatric hospitals.Methods:Totally 1 365 family members of inpatients with mental disorders were-surveyed,and were assessed with the Perceived Devaluation-Discrimination Scale and a self-made demographic characteristics questionnaire.Results:The screening rate of stigma among the patient's family members was 61.5％.Males(OR=2.26,95％CI:1.06-5.01),age group of 18-29 years(OR=1.91,95％CI:1.15-3.20),monthly income ≥500 yuan(P＜0.05),disease duration of 0.5-＜lyear(OR=3.14,95％CI:1.66-6.03),care for patients within＜lyear(P＜0.05),teachers(OR=2.32,95％CI:1.24-3.44),self-employed person(OR=1.63,95％CI:1.02-2.24),civil servants(OR=1.77,95％CI:1.09-2.45),schizophrenia(OR=1.87,95％CI:1.32-2.42),affective disorders(OR=1.52,95％CI:1.03-2.016)were the main riskfactors of stigma.Conclusions:Family members of patients with mental disorders generally have a severe stigma,especially of patients with schizophrenia and affective disorders.

OBJECTIVE： To separate and identify chemical components in ethyl acetate fraction and water fraction from Tripterygium wilfordii， and to provider basis for further pharmacological study. METHODS： The ethyl acetate fraction and water fraction from T. wilfordii were separated and purified by MCI GEL-CHP 20P column chromatography， C18 RP silica gel column chromatography， Sephadex LH-20 gel column chromatography and HPLC. The structures of compounds were analyzed and identified by 1H-NMR， 13C-NMR and physicochemical properties. RESULTS： Two compounds were isolated from ethyl acetate fraction of T. wilfordii， namely orthosphenic acid （compound 1）， dibutylphthalate （compound 2）. Eight glucosides were isolated from water extract of T. wilfordii， namely 2，6-dimethoxy-4-hydroxymethyl-phenyl-1-O-beta-D-glucopyranoside （compound 3）， 2，6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucoside（compound 4）， 4-hydroxy-1-（2-hydroxyethyl）-phenyl-3-O-β-D-glucopyranoside （compound 5）， 3，4-dimethoxy-phenyl-1-O-β-D-glucopyranoside （compound 6），β-adenosine （compound 7）， ligustrin （compound 8）， epicatechin-8-C-β-D-galactoside （compound 9） and 2-hydroxynaringenin-7-O-β-glucoside （compound 10）. CONCLUSIONS： Chemical components of ethyl acetate fraction and water fraction are separated and identified from T. wilfordii.

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.
Animals ; Blotting, Western ; Cathepsin B ; drug effects ; metabolism ; Cathepsins ; drug effects ; metabolism ; Cell Line, Tumor ; drug effects ; Colorectal Neoplasms ; blood supply ; genetics ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; methods ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; drug effects ; metabolism ; Integrin alpha Chains ; drug effects ; metabolism ; Neovascularization, Pathologic ; genetics ; Receptors, Urokinase Plasminogen Activator ; drug effects ; metabolism ; STAT3 Transcription Factor ; drug effects ; metabolism ; Signal Transduction ; Sphingomyelin Phosphodiesterase ; drug effects ; metabolism ; TOR Serine-Threonine Kinases ; drug effects ; metabolism ; Tumor Suppressor Protein p53 ; drug effects ; metabolism ; Up-Regulation ; Vascular Endothelial Growth Factor A ; drug effects ; metabolism

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.
Animals ; Apigenin ; Blotting, Western ; Cell Cycle ; Cell Division ; Cell Proliferation ; drug effects ; genetics ; Colorectal Neoplasms ; Cyclin-Dependent Kinase Inhibitor p21 ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Ginsenosides ; Glycyrrhizic Acid ; Humans ; Lactones ; Phosphorylation ; genetics ; RNA, Messenger ; Saponins ; Sesquiterpenes ; Signal Transduction ; TOR Serine-Threonine Kinases ; drug effects ; Triterpenes ; Tumor Suppressor Protein p53 ; drug effects

The first rate-limiting enzyme of the serine synthesis pathway (SSP), phosphoglycerate dehydrogenase (PHGDH), is hyperactive in multiple tumors, which leads to the activation of SSP and promotes tumorigenesis. However, only a few inhibitors of PHGDH have been discovered to date, especially the covalent inhibitors of PHGDH. Here, we identified withangulatin A (WA), a natural small molecule, as a novel covalent inhibitor of PHGDH. Affinity-based protein profiling identified that WA could directly bind to PHGDH and inactivate the enzyme activity of PHGDH. Biolayer interferometry and LC-MS/MS analysis further demonstrated the selective covalent binding of WA to the cysteine 295 residue (Cys295) of PHGDH. With the covalent modification of Cys295, WA blocked the substrate-binding domain (SBD) of PHGDH and exerted an allosteric effect to induce PHGDH inactivation. Further studies revealed that with the inhibition of PHGDH mediated by WA, the glutathione synthesis was decreased and intracellular levels of reactive oxygen species (ROS) were elevated, leading to the inhibition of tumor proliferation. This study indicates WA as a novel PHGDH covalent inhibitor, which identifies Cys295 as a novel allosteric regulatory site of PHGDH and holds great potential in developing anti-tumor agents for targeting PHGDH.