Identification of a novel PHGDH covalent inhibitor by chemical proteomics and phenotypic profiling

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.

Diagnostic value of transbronchial lung biopsy in pulmonary alveolar proteinosis / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the diagnostic value and limits of transbronchial lung biopsy (TBLB) in pulmonary alveolar proteinosis (PAP).@*METHODS@#The complete hospital data from Second Xiangya Hospital, Central South University, between June, 2006 and December, 2012, were analyzed retrospectively in 25 patients with PAP (who were diagnosed pathologically by TBLB or not by TBLB) and in 4 patients with other disease (who were misdiagnosed by TBLB).@*RESULTS@#Among the 25 patients with PAP, 14 patients were confirmed by TBLB in the fi rst time [TBLB positive rate in the first time was 56% (14/25)]; 6 patients who were misdiagnosed by TBLB in the fi rst time were confirmed by TBLB in the second time [the positive rate in the second time was 24% (6/25)]. Th e total positive rate was 80% (20/25). Th e total negative rate was 20% (5/25). Five patients with PAP, who showed negative results in TBLB analysis, were confirmed by the typical CT as well as the whole lung lavage. In addition, 4 patients with other diseases were misdiagnosed as PAP by TBLB.@*CONCLUSION@#TBLB is a very good diagnosis method. But the negative results in the first time cannot exclude PAP. TBLB should be repeated if conditions allow. If clinical manifestation and CT results are typical, it can be used for diagnosis of PAP combined with the positive results from bronchoalveolar lavage fluid check. However, for non-typical cases, it needs TBLB pathological diagnosis.

Simultaneous determination of major coumarins′ content in processing Peucedani Radix materials by method of substitute for reference substance / 中国药科大学学报

@#(±)-Praeruptorin A(PA)was used as the standard reference substance to establish a new method of substitute for reference substance. The contents of both(±)-praeruptorin B(PB)and(+)-praeruptorin E(PE)were calculated by the method. The major content of coumarins in different processing materials could be gained by determining the content of(±)-praeruptorin A, which could provide scientific evidence for the research on pharmacology discrepancy and quality standards. The method was carried out on a Welch Materials Ultimate XB-C18 column(4. 6 mm×250 mm, 5 μm), with a gradient mobile phase of methanol and water at the flow rate of 1. 0 mL/min. The column temperature was 30 °C and detection wavelength was set at 322 nm. According to this method, the relative correlation factors(f)of(±)-praeruptorin B and(+)-praeruptorin E were determined as 0. 787 2 and 0. 969 0, respectively. After determine the contents of 15 batches of materials from different sources, no significant differences between substitute method and external standard method were observed. It was a feasible and credible method to determine the content of major coumarins in Peucedani Radix with different processing materials. The results showed lower contents of coumarins in processing materials than in raw materials.

Crazy paving pattern caused by pulmonary alveolar proteinosis: CT findings and the pathologic basis / 中南大学学报(医学版)

OBJECTIVE@#To explore CT findings and pathologic basis of crazy paving pattern caused by pulmonary alveolar proteinosis.@*METHODS@#Twenty-four patients who were diagnosed pathologically as pulmonary alveolar proteinosis by transbronchial lung biopsy and bronchoalveolar lavage fluid from June 2006 to May 2012 were included in this retrospective study. All patients underwent a 64-slice CT of the lungs.@*RESULTS@#CT findings: crazy paving pattern was observed on CT imaging of all 24 patients. In 23 patients, crazy paving pattern displayed strip-shaped opacities with smooth edges, and there was a clear boundary between the pathological and normal lung tissues. The reticular opacities were connected with peripheral blood vessels and the branches were formed, and their diameters decreased slightly. Microscopically, hemangiectasis were seen in 17 patients.@*CONCLUSION@#Crazy paving pattern caused by pulmonary alveolar proteinosis displayed clear edges, and smooth reticular opacities, most of which were due to hemangiectasis of interlobular, interacinar and interalveolar septa. These findings of CT are helpful for the specific diagnosis of pulmonary alveolar proteinosis.

Lung MRI at 3T：comparison of CT and MRI in initial evaluation of pulmonary alveolar proteinosis / 中南大学学报(医学版)

Objective:To explore whether the phospholipidoproteinaceous material deposit within the alveoli by a high-ifeld 3T MRI has signal characters and its application for diagnosing pulmonary alveolar proteinosis.Methods:A total of 11 patients with pulmonary alveolar proteinosis previously diagnosed by ifberoptic bronchoscope lung biopsy underwent 64-slice helical CT scans and 3T MRI scans, and the CT scans and the MRI scans were compared. Results:hTe phospholipidoproteinaceous material deposit within the alveoli presented longer or equal T1 relaxation time and longer T2 relaxation time, without characters of fatty or deposits of protein-like substance signals and enhancement. The distribution, form, number and size of the lesions at T2WI were almost the same as those at CT, the lesions were irregular in morphology, and there was a clear boundary between the lesions and the adjacent normal lung tissues. Dynamic contrast-enhanced MRI showed thickened pulmonary arteriolae and venulae in the lesions with more obviously thickened pulmonary venulae, which were in conformity with the pulmonary artery and venule enhancement. CT scan in 1 out of the 11 cases showed lesions in both lungs mainly consisted of stripe-shaped and reticular structures, and no obvious sign of pulmonary alveolar proteinosis residue was found. MRI scan detected alveolar proteinosis that failed to be shown by CT scan. Conclusion:3T MRI T2WI can easily detect the lesions of long T2 signals formed by the lipoproteinaceous material deposit within the alveoli. In the lesions, geographic appearance was presented, and the crazy paving pattern was dimly visualized. MRI can relfect the morphological characters of PAP like CT and it is slightly better compared with CT in such aspects as evaluating the theraputic effect of lung lavage. As supplement to CT, high-field 3T MRI can serve as an important examination for lung diseases.

Characteristic CT findings and pathologic basis of ground glass opacity caused by pulmonary alveolar proteinosis / 中南大学学报(医学版)

OBJECTIVE@#To explore characteristic CT findings and pathologic basis of ground glass opacity caused by pulmonary alveolar proteinosis (PAP).@*METHODS@#Retrospective analysis of CT and pathological findings of 24 patients with PAP who were pathologically diagnosed from June 2006 to August 2011.@*RESULTS@#Findings with CT: the lesions of the 24 patients mainly presented ground glass opacities. Local consolidations were seen in 8 patients. In 23 patients part of ground glass opacities bordered strip-shaped opacities with smooth edges, and there was a clear boundary between them and the bordering normal lung tissues, presenting a geographic appearance. Lesions in the 5 cases were mixed with alveoli or lobule aerocele, which made ground glass opacities present curved edges. Crazy paving pattern was detected in the 24 patients. Microscopically, the alveoli were seen to be filled with floccules proteinaceous material in various quantities in the 24 patients; hemangiectasis and congestion were seen in 17 patients, and enlarged alveolar cavities were seen in 5 patients.@*CONCLUSION@#PAP usually causes ground glass opacities with clear edges, and different from ground glass opacities with obscure edges caused by other pulmonary diseases. They are relatively specific to the imagining diagnosis to PAP.

Correlation between cerebral microbleed and hemorrhage transformation after urokinase intra-arterial thrombolysis in patients with acute ischemic stroke: a prospective case series study / 国际脑血管病杂志

Objective To investigate the correlation between cerebral microbleed (CMB) and hemorrhage transformation (HT) after urokinase intra-arterial thrombolysis in patients with acute ischemic stroke.Methods The patients with acute cerebral infarction treated with intra-arterial urokinase were enrolled.They were divided into either an HT group or a non-HT goup according to whether they had HT or not.Conventional MRI sequences,susceptibility-weighted imaging (SWI),and CT scan were performed before procedure.CT or MRI was reexamined within 48 hours after procedure.The patients' demographic data,vascular risk factors,and the SWI sequences showed the numbers of CMB were documented in detail and they were compared and analyzed.Results A total of 62 patients were included,22 in HT group and 40 in non-HT group.Univariate analysis showed that the proportions of hypertension (81.8％ vs.57.5％ ; x2 =3.125,P =0.048),diabetes (63.6％ vs.40.0％ ; x2 =4.019,P =0.042),smoking (72.7％ vs.37.5％ ; x2 =4.971,P =0.030),and presence of CMB (x2 =5.297,P =0.018) of patients in the HT group were significantly higher than those in the non-HT group.Multivariate logistic regression analysis showed that hypertension (odds ratio [OR]1.51,95％ confidence interval [CI]1.102-2.954; P =0.028),diabetes (OR 1.48,95％ CI 1.09-2.825; P =0.039),and CMB (OR 1.867,95％ CI 1.103-3.158; P =0.020) were the independent risk factors for HT after urokinase intra-arterial thrombolysis in patients with acute ischemic stroke.Conclusions CMB was one of the independent risk factors for occurring HT after urokinase intra-arterial thrombolysis in patients with acute ischemic stroke.

Triterpenoid Saponins from Roots of Gypsophila pacifica / 中草药·英文版

Objective To study the chemical constituents from the roots of Gypsophila pacifica. Methods The chemical constituents were isolated by various column chromatographic methods and their structures were identified by spectral data together with physicochemical analysis. Results Five compounds were isolated and identified as 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl gypsogenin 28-O-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (1), 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (2), 3-O-β-D-galactopyranosyl-( 1→ 2 )-[β-D-xylopyranosyl-( 1→ 3 ) ]-β- D-glucuronopyranosyl gypsogenin 28-O-β-D-glucopyranosyl-( 1→ 3 )-[β-D-xylopyranosyl-( 1→4 ) ]-α-L-rhamnop yranosyl-( 1 → 2 )-β-D-fucopyranoside (3),3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-β-D-glucopyranosyl-( 1→ 3 )-[β-D-xylopyranosyl-( 1→4 ) ]-α-L-rhamnopyranosyl-( 1→2 )-β-D-fucopyranoside ( 4), 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-α-L-arabinopyranosyl-(1→4)-α-L-arabinopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (5). Conclusion The five compounds are isolated from this plant for the first time.

Analysis of bioactive flavonoids and resources research of main Herba Epimedii species from different origins / 中国药科大学学报

Herba Epimedii (named Yinyanghuo in Chinese) of the family Berberidaceae is one of the most famous traditional Chinese medicines (TCMs) recorded in the pharmacopoeia of the People's Republic of China.Total flavonoids of Epimedium (TFE) are considered to be its active components.TFE content was measured by ultraviolet method and its representative constituents,icariin and epimedin C,were determined by high performance liquid chromatography.The test results from 36 samples of 4 species showed that the contents differed significantly in different species.The average data of the summation (icariin and epimedin C) content were 20.83,7.61,14.43,23.29 mg/g in Epimedium pubescens Maxim.,E.koreanum Nakai.,E.brevicornum Maxim.,E.sagittatum (Sieb.Et Zucc.) Maxim.,respectively.Both E.sagittatum and E.pubescens performed better than the other two batches.E.sagittatum hardly has any circulation in Chinese herbal medicine market.The results showed that E.pubescens,which had much more amounts of icariin,epimedin C and TFE than those of other species,has a better quality and may be considered as potential anti-osteoporosis drug.

HPLC-ESI-MS(n) analysis of chemical constituents in Semen Ziziphi Spinosae / 中国中药杂志

<p><b>OBJECTIVE</b>To study on the chemical constituents of effective part of Semen Ziziphi Spinosae.</p><p><b>METHOD</b>The sample was extracted with methanol and purified by macroporous resin. The structures were identified by high performance liquid chromatography/electrospray ionization with multi-stage tandem mass spectrometry.</p><p><b>RESULT</b>Thirteen compounds were identified from the effective part of Semen Ziziphi Spinosae.</p><p><b>CONCLUSION</b>The method is simple and rapid for the identification of the flavonoids and triterpenoid saponins from Semen Ziziphi Spinosae.</p>

Effect of preoperative transcatheter arterial chemoembolization on apoptosis of hepatocellular carcinoma cells / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the effect of preoperative transcatheter arterial chemoembolization(TACE) on apoptosis of hepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>A total of 136 patients with HCC underwent liver resection. One to five courses of TACE prior to liver resection were performed in 79 patients (TACE group), in which one to four courses of chemotherapy alone were performed in 11 patients (group A), one to five courses of chemotherapy combined with iodized oil were performed in 33 patients (group B), one to three courses of chemotherapy combined with iodized oil and gelatin sponge were performed in 23 patients group C) and one to three courses of chemotherapy combined with iodized oil, ethanol and gelatin sponge were performed in 12 patients (group D). The other 57 patients only received liver resection (non-TACE group). The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining. The expressions of Bcl-2 and Bax protein were detected by immunohistochemical method.</p><p><b>RESULTS</b>The apoptotic index(AI) and level of Bax protein in HCC cells were significantly higher in groups A, B, C and D than those in the non-TACE group (P < 0.05). The level of Bcl-2 protein and ratio of Bcl-2 to Bax protein of HCC cells were significantly lower in Groups A, B, C and D than those in the non-TACE group (P < 0.05).</p><p><b>CONCLUSION</b>Preoperative TACE regimens may enhance apoptosis of HCC cells by up-regulating the expression of Bax protein and down-regulating the expression of Bcl-2 protein and ratio of Bcl-2 to Bax protein expression.</p>

Digital Subtraction Angiography Characteristics of Traumatic Renal Hemorrhage / 实用放射学杂志

Objective To study DSA characteristics of traumatic renal hemorrhage and to discuss the methods for treatment of this disease.Methods Superselective renal arterial angiography was carried in 14 patients with traumatic renal hematuria from 2001~2007.11 cases were iatrogenic renal damage and 3 cases were renal trauma.6 of them were under unstable hemodynamic conditions.Results The bleeding was found in 7 cases,located at the interlobar artery,the branch of segmental artery of the kidney,and the proximal end of arcuate artery.They presented as renal arterial-pelvic fistula(4 cases),arterial venous fistula(1 case) and renal arterial-pelvic fistula with AV fistula(2 cases),2 of them accompanied with pseudoaneurysm.The characteristics on DSA were radiopaque contrast medium overflow rapidly and abundantly,and the lesions were far from peripheral area of kidney,in the other 7 patients,the bleeing arteriae were located at the distal end of arcuate artery and the smaller branches of interlobular artery.The radiopaque material remained in parenchyma of kidney until parenchymal phase.There were no obvious artery-vein fistula or renal arterial-pelvic fistula in all cases.Conclusion DSA examination is very helpful in diagnosis and treatment of traumatic renal hemorrhage.