Multicellular spheroid (MCS) can simulate many aspects of the in vivo physiological and pathological conditions in many aspects,better reflect the in-vivo behavior of cells in tumors.So it's increasingly accepted as a valuable tool for evaluating the efficacy of therapeutic intervention including chemotherapy,radiotherapy,immunotherapy and combined therapy.Various spheroid co-culture approaches have been presented to study heterologous cells interaction in solid tumors.The present review briefly introduces the methodology and applications of MCS with focus on the up-to-date information.

Objective: To establish an accelerated solvent extraction(ASE)-HPLC method to determine chrysophanol and aurantio-obtusin in Cassia obtusifolia L.Methods: The optimal extraction conditions were defined by orthogonal tests using ASE.The method was carried out on an ACE Excel C18-PFP column (75 mm×2.1 mm,2.5 μm) with the mobile phase consisting of 0.1% phosphoric acid solution-acetonitrile with gradient elution.The column temperature was 40 ℃,the flow rate was 0.4 ml·min-1, and the detection wavelength was 284 nm. Results: The best process parameters of ASE were as follows:the extraction solvent was methanol, the extraction temperature was 120 ℃ and the static extraction duration was 5 minutes for three cycles.The ASE method needed only 1/9 of the time as the pharmacopoeia method,while the extraction efficiency of the ASE method was higher.The linear ranges of cassia obtusifolia L.and Chrysophanol were at 0.73~58.57 μg·ml-1(r=0.999 7) and 1.09~87.29 μg·ml-1(r=0.999 6).The average recoveries were 102.7%(RSD=0.8%) and 98.2%(RSD=1.5%).Conclusion: The method is simple, rapid and sensitive, which can be used for the rapid determination of aurantio obtusin and chrysophanol in Cassia obtusifolia L.

Tropane alkaloids,such as hyoscyamine (or its more stable racemate atropine) and scopolamine,are remembered as the oldest drugs in medicine with wide pharmaceutical applications clinically for their mydriatic,antispasmodic,anticholinergic,analgesic and sedative properties.Presently,the supply of tropane alkaloids are entirely rested on their isolation from plant materials,yet the low contents of tropane alkaloids in plants raise the requirement to breed new varieties of high-yield tropane alkaloids and improving the production of tropane alkaloids in hairy root cultures or plants by biotechnology has be a research focus in the field of secondary metabolism.For the past decade,there have been important progresses on biosynthesis of tropane alkaloids and their molecular biology,some new pathway genes have been identified and many achievements have also been acquired in metabolic engineering of tropane alkaloids.This review summarized the recent advances in above aspects,and the problems and orientations of future research are also discussed and proposed.

Objective To improve the diagnostic accuracy of ovarian endometrioma and to discriminate it from other adnexal masses by analyzing its CT manifestations. Methods The CT images of 57 patients with 77 ovarian endometriomas proved by surgery and pathology were studied retrospectively.Results The cysts ranged in diameter from 2 to 14 cm with the mean size of 6.2 cm?5.1 cm. The margins of the cyst were sharp in 39 and ill defined in 38 endometriomas. Thin and thickened walls were found respectively in 40 and 37 of 77 endometriomas. Unilocular, bilocular and multilocular forms were seen respectively in 29(37.7%), 23(29.9%) and 25(32.5%) of 77 endometriomas. Gourd and sister forms were present respectively in 15 and 8 of 23 bilocular cysts, and sister loculi, outer son loculi, inner son loculi, and mixed forms in 10, 9, 1 and 5 of 25 multilocular endometriomas. The density of the cyst fluid varied from 11～65 HU with the mean of 28.5 HU. New hemorrhage inside cyst was found in 6 cysts. Slight or intermediate enhancement of the cyst wall of endometriomas in 42, and significant enhancement in 35 were demonstrated. 54 cysts, 13 cystadenomas with 18 lesions and 4 malignant tumors with 5 lesions were diagnosed by preoperatively by CT. The accurate rate of CT characterization was 71.4%. Conclusion The certain morphologic characteristics of the ovarian endometrioma can be showed by CT in higher extent. It is critical to familiar with these characteristics to improve the diagnostic accuracy, and to avoid misdiagnosis.

Objective To analytically screen the volatile components from Chenxiang Huaqi Tablets (CHT) using gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) combined with automated mass spectral deconvolution, and establish the methods of multiple components contents (camphor, bornyl acetate, and patchouli alcohol) from CHT using multi reaction monitor (MRM). Methods The chromatographic column was HP-5MS (30 m × 0.32 mm × 0.25 μm). Temperature program: the initial temperature was 100 ℃, at 15 ℃/min up to 300 ℃ for 2 min. Injection port temperature was 280 ℃; Ionization methods: EI ion source with ion source temperature of 230 ℃; Quadrupole temperature was 150 ℃; Interface temperature was 280 ℃; Inject volume was 1 μl. Results A total of thirteen components were screened by automated mass spectral deconvolution soft. Under the conditions of MRM collection, camphor, bornyl acetate, and patchouli alcohol were in the range of 4.5-90 mg/mL (r = 0.999 8), 3.3-66 mg/mL (r = 0.999 8), and 2.6-51.5 mg/mL (r = 0.999 9), respectively, the ratio of each component concentration to its peak area was linear, and the average recovery (n = 6) was 101.15%, 102.64%, and 100.10% respectively; The mass fraction of 10 batches of samples were in the ranges of 0.278-0.311, 0.381-0.438, and 0.229-0.381. Conclusion The method is accurate, simple, and good repeatability. It can be used for the simultaneous determination of camphor, bornyl acetate, and patchouli alcohol in CHT, which provides a reference for the improvement of the quality of this variety.

Esophageal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Melanoma ; metabolism ; pathology ; Melanoma-Specific Antigens ; metabolism ; Middle Aged ; S100 Proteins ; metabolism

OBJECTIVETo study the molecular mechanism of extracts from Euodia rutaecarpa on hepatotoxicity in mice.

METHODTotally 30 KM mice were divided into 3 groups and orally administrated extracts from E. rutaecarpa for consecutively 15 days. The expressions of Erkl/2, CDK8, CK1e, Stat3 and Src were detected by Western blotting method.

RESULTThe extracts from E. rutaecarpa could up-regulated Erkl/2, CDK8 and CK1e expressions (P <0.01) and down-regulate Stat3 and Src (P <0.01).

CONCLUSIONThe molecular mechanism of E. rutaecarpa on hepatotoxicity may be correlated with Erkl/2, CDK8, CKle, Stat3 and Src signal molecules.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Down-Regulation ; Drugs, Chinese Herbal ; toxicity ; Evodia ; chemistry ; Female ; Fruit ; chemistry ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; metabolism ; Male ; Mice ; Plants, Medicinal ; Signal Transduction ; drug effects ; Specific Pathogen-Free Organisms ; Triglycerides ; blood ; Up-Regulation

Objective To evaluate mivacurium-induced release of histamine in the patients undergoing general anesthesia.Methods Eighty patients of both sexes,aged 19-58 yr,weighing 42-75 kg,of ASA physical status Ⅰ or Ⅱ,undergoing ureteroscopy under general anesthesia,were randomly divided into 2 groups (n =40 each) using a random number table:rocuronium group (group R) and mivacurium group (group M).Anesthesia was induced with iv midazolam 0.04 mg/kg,fentanyl 3 μg/kg,etomidate 0.3 mg/kg,and mivacurium 0.20 mg/kg (group M) or rocuronium 0.75 mg/kg (group R).Streamlined liner of the pharynx airway (SLIPA) was inserted for mechanical ventilation.PET CO2 was maintained at 35-40 mmHg.Anesthesia was maintained with fentanyl and propofol.Before muscle relaxant administration,at 3 min after muscle relaxant administration and at 5 min after insertion of SLIPA,venous blood samples were collected for determination of plasma histamine concentrations (by HPLC) and the histamine release was classified.Results Compared with group R,the plasma histamine concentration and histamine release were increased at 3 min after muscle relaxant administration in group M.No fatal responses caused by histamine release were found in the two groups.Conclusion Although mivacurium induces release of histamine,it can still be safely used for the patients undergoing ureteroscopy under general anesthesia.

Objective To evaluate mesenteric muhidetector row CT angiography (MDCTA) in the diagnosis of acute mesenteric ischemia (AMI).Methods In this study,43 cases of AMI proven by clinical criteria,or operation and pathology underwent whole abdomen MDCT precontrast,arterial phase and venous phase scan with 0.6 mm collimation.The mesenteric arteries and veins were reconstructed by using volume rendering (VR),maximum intensity projection (MIP),thin slab maximum intensity projection (TSMIP) techniques,and abnormal CT angiography findings as well as abnormal bowel and mesentery were analyzed.Results It was found that AMI was caused by superior mesenteric artery (SMA) embolism (n =4),SMA thrombosis (n = 6),mesenteric and portal venous thrombosis (n = 13),SMA dissection (n =5),strangulated bowel obstruction (n = 10) and vasculitis (n = 5).MDCTA showed clearly the position,shape,severity and extent of the vascular occlusion,narrow and dissection.It could also demonstrate the abnormal course and direction of the vessels including vascular gathering,shift,tortuosity,retortion,and twist.Furthermore,the pathogenesis of various conditions including atherosclerotic plaque,emboli,thrombosis,dissection,tumorous invasion,strangulated bowel obstruction and vasculitis could be identified by MDCTA.Conclusion MDCTA can clearly demonstrate direct signs leading to AMI,and identify AMI and its etiology at early stage,with the combination with its indirect findings.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.