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.

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.

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.

OBJECTIVE: To observe the effects of "" acupuncture on cerebral blood flow in high-risk patients of cerebral ischemic stroke based on arterial spin labeling (ASL) and perfusion-weighted imaging (PWI), and to evaluate the clinical efficacy. METHODS: A total of 180 patients with transient ischemic attacks (TIA) / minor ischemic stroke (MIS) were randomly divided into an acupuncture A group, an acupuncture B group and a medication group, 60 cases in each group. The patients in the acupuncture A group were treated with "" acupuncture at Baihui (GV 20), Fengfu (GV 16), Yamen (GV 15), Dazhui (GV 14), Shenzhu (GV 12), Zhiyang (GV 9), Mingmen (GV 4), Yaoyangguan (GV 3) and Jingjiaji (EX-B 2), once a day; the patients in the acupuncture B group were treated with identical acupoints but was given once every other day; the patients in the medication group were treated with oral administration of nimodipine tablets, 30 mg, three times daily. All the three groups were treated for four weeks. ASL and PWI, including relative cerebral blood volume (rCBV), relative cerebral blood flow (rCBF), relative mean transit time (rMTT) and relative time to peak (rTTP), were conducted before and after treatment; the changes of the test indexes were compared before and after treatment. The clinical efficacy of the three groups was compared. RESULTS: Compared before treatment, the numbers of ASL normal perfusion in the 3 groups were significantly increased after treatment (all <0.01); the number of ASL normal perfusion in the acupuncture A group was higher than that in the acupuncture B group (<0.05), but was not significantly different from that in the medication group (>0.05). Compared before treatment, rCBV and rCBF in the 3 groups were significantly increased after treatment (all <0.01), and rMTT and rTTP were significantly reduced (all <0.01). After treatment, rCBV and rCBF in the acupuncture A group were higher than those in the acupuncture B group (all <0.05); the rMTT and rTTP in the acupuncture A group were lower than those in the acupuncture B group (all <0.05); the differences of PWI parameters after treatment were not statistically significant between the acupuncture A group and medication group (all >0.05). The total effective rate was 88.3% (53/60) in the acupuncture A group, 73.3% (44/60) in the acupuncture B group and 90.0% (54/60) in the medication group; the total effective rate in the acupuncture A group was superior to that in the acupuncture B group (<0.05), but was not significantly different from that in the medication group (>0.05). CONCLUSION "" acupuncture could effectively improve the hypoperfusion of cerebral blood flow in patients with high risk of cerebral ischemic stroke, reduce the incidence of severe CIS; acupuncture for once a day is better than once every other day.
Acupuncture Therapy ; Brain Ischemia ; prevention & control ; Cerebrovascular Circulation ; Humans ; Risk Factors ; Stroke

Objective To discuss the techniques and methods of surgery for brain gliomas located in eloquent areas at awake anesthesia. Methods Nineteen patients with brain gliomas in eloquent areas, admitted to our hospital from December 2014 to May 2017, were operated under awake anesthesia with neuronavigation and intraoperative ultrasonography for locating the lesions and intraoperative direct electrical stimulation for functional mapping of the eloquent areas. All patients were followed up from 3 to 18 months; the surgical efficacies were analyzed. Results Of 19 patients, 18 (94.74％) were achieved awake and alert during brain mapping and resection of the tumors;17 (89.47％) were detected the motor areas by intraoperative direct electrical stimulation, 6 (31.58％) were detected the sensory cortex and 12 (63.16％) were detected language related cortex. Of 19 patients, MR imaging 2-3 months after surgery indicated that 5 (26.32％) received total resection of lesions, 9 (47.37％) subtotal resection of lesions and 5 (26.32％) partial resection of lesions. Seven patients (36.84％) had transitory postoperative aphasia, 4 (21.05％) were with transitory postoperative dyskinesia and one (5.26％) with permanent dyskinesia. Conclusion Comprehensive applications of awake anesthesia, neuronavigation, intraoperative ultrasonography and intraoperative direct electrical stimulation technologies allow maximum safe resection of gliomas in eloquent areas and protection of brain function.

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

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.

Objective To explore the dignostic value of the candidate molecular markers in Urothelium carcinoma based on Illumina Transcriptome Sequencing. Methods Candidate genes which were screened with transcriptome sequencing were validated by using real-time fluorescent quantitative PCR in 31 pairs of tumor and normal tissues of Urothelium carcinoma patients from Affliated Hai kou Hospital Xiangya School of Medicine Central South University. Results The four candidate genes (CDH1,VEGFA,PTPRF and CLDN7) in tumor tissues were up-regulated compared with normal groups in samples of sequencing. The relative expression of VEGFA mRNA were higher than that in normal tissues. There were significant difference in two groups (P<0.05). PTPRF and CLDN7 were also up-regulated, but no significant difference was found (P > 0.05). PTPRF was closely related with the recurrence of tumor (P=0.002), and the predict sensitivity and speciality rate were 90.0%and 83.3%respectively. Conclusions CDH1 and VEGFA play important roles in the occurrence and development of bladder urothelial carcinoma, and they may be the possible biomarkers. PTPRF is expected to be a molecular reference recurrent predictor of bladder tumor.

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 .