[Study on the assignment method of intensity modulated radiotherapy plan for nasopharyngeal carcinoma based on MR images].

Objective: To discuss the influence of different computed tomography (CT) value assignment methods on dose calculation of intensity modulated radiotherapy (IMRT) plan which designed for nasopharyngeal carcinoma (NPC) and the value assignment methods of IMRT plan for NPC based on magnetic resonance (MR) images. Methods: Simulation CT and MR image of 32 NPC patients in Shandong Cancer Hospital from March 2018 to November 2018 were selected for this study. Populate CT values were obtained by contouring and analyzing the simulation CT of patients' tissue, including bone, air, brain, eyeball, optic-nerve, lens, parotid, masseter, skin. Pseudo-CT were generated by different CT value assignment methods: CT1: CT value of all tissues was set to 0HU; CT2: CT value of air cavity was set to populate CT value based on CT1; CT3: CT value of Bone was set to populate CT value based on CT2; CT4: CT value of each soft tissue were set to populate CT value based on CT3. The IMRT plan for NPC as Plan0 was designed base on simulation CT. Then Plan0 was transplanted to four pseudo-CT to recalculate the dose and obtain Plan1, Plan2, Plan3 and Plan4, the differences of dosimetric parameters were compared with Plan0. NPC-IMRT plan was designed base on MR images by using the assignment method with CT value of each tissue were set to populate CT value. Results: In the head and neck CT images, the average populate CT values of bone and cavity were 621 HU and -720 HU, respectively. The populate CT values of other soft tissue ranges from -20 HU to 70 HU. The differences of dosimetric indexes of Plan1, Plan2, Plan3, Plan4 decreased sequentially compare to Plan0, the difference of the dosimetry parameters of Plan4 and Plan0 was the smallest. The differences of PTV D(99), PTV D(95), isocenter dose, D(mean) of all tissues, D(max) of bilateral eye balls, D(max) of bilateral lens, D(max) of bilateral optic nerves, D(mean) of bilateral parotid, V(20) of bilateral parotid, D(50) of bilateral parotid, D(max) of spinal cord, D(max) of brainstem, D(5) of brainstem between Plan4 and Plan0 were all less than 1%. The difference of V(30) in bilateral parotid between Plan4 and Plan0 was less than 1.5%. In the comparison of the pixel dose distribution, the regions of dose distribution difference greater than 1% mainly distributed in the air cavity, bone periphery and the skin. The target area of the IMRT plan for NPC based on MR images met 95% of the prescribed dose, and the dose of each organ at risk was within the dose limit. Conclusions: The assignment method of each tissue and organs set to populate CT value compared with other methods has the least influence on the dose calculation of NPC-IMRT plan, which could meet the clinical requirements. Therefore, it should be the first choice of assignment method when designing NPC-IMRT plan based on MR image.

Oncogenic activation of JAK3-STAT signaling confers clinical sensitivity to PRN371, a novel selective and potent JAK3 inhibitor, in natural killer/T-cell lymphoma.

Aberrant activation of the JAK3-STAT signaling pathway is a characteristic feature of many hematological malignancies. In particular, hyperactivity of this cascade has been observed in natural killer/T-cell lymphoma (NKTL) cases. Although the first-in-class JAK3 inhibitor tofacitinib blocks JAK3 activity in NKTL both in vitro and in vivo, its clinical utilization in cancer therapy has been limited by the pan-JAK inhibition activity. To improve the therapeutic efficacy of JAK3 inhibition in NKTL, we have developed a highly selective and durable JAK3 inhibitor PRN371 that potently inhibits JAK3 activity over the other JAK family members JAK1, JAK2, and TYK2. PRN371 effectively suppresses NKTL cell proliferation and induces apoptosis through abrogation of the JAK3-STAT signaling. Moreover, the activity of PRN371 has a more durable inhibition on JAK3 compared to tofacitinib in vitro, leading to significant tumor growth inhibition in a NKTL xenograft model harboring JAK3 activating mutation. These findings provide a novel therapeutic approach for the treatment of NKTL.

TRAV gene expression in PBMCs and TILs in patients with breast cancer analyzed by a DNA melting curve (FQ-PCR) technique for TCR α chain CDR3 spectratyping.

PURPOSE: To explore the expression of the TRAV gene in peripheral blood mononuclear cells (PBMCs) and in tumor-infiltrating lymphocytes (TILs) in the patients with breast cancer using a DNA melting curve (FQ-PCR) technique for T cell receptor (TCR) alpha chain CDR3 spectratyping. Peripheral blood samples and tissue samples were obtained from thirty breast cancer patients. Total RNA was extracted from PBMCs and tumor tissues and then reverse transcribed into cDNA. FQ-PCR was used to amplify the human TCR alpha chain CDR3 region with the primers to the TRAV and TRAC genes. TCR alpha chain CDR3 spectratyping and partial CDR3 sequencing were used to determine use of TRAV gene product in T cell responses. TCR alpha CDR3 spectratyping showed preferential usage of certain TRAV genes in the PBMCs and TILs of all patients with breast cancer. The frequencies of TRAV1.1, TRAV9, and TRAV29 exceeded 30% in PBMCs and the frequencies of TRAV1.1 and TRAV22 exceeded 30% in TILs. More than three quarters of the patients (23/30) overexpressed the same gene in both PBMCs and TILs; for example, patient-1 highly expressed TRAV9 in the PBMCs and TILs. Patients with positive or negative tumor markers of estrogen receptor (ER), progesterone receptor (PR), pS2, C-erbB-2, nm23, P53, and Ki-67 showed no significant common TRAV gene expression, but some TRAV gene preferential usage frequencies exceeded 20%. For example, five of seven patients positive for ER had high levels of expression of TRAV1.1 and TRAV3. Finally, the amino acid sequence of TCR CDR3 region showed some common motifs in some of the patients. CONCLUSIONS: TRAV gene expression was complex and diverse in the patients with breast cancer. The TRAV gene usage may be closely related to the diversity of breast tumor antigens and the differential immune responses observed in individual patients. Research into the immunological mechanism of T cells may provide guidance for individual T cell-directed therapy for breast cancer.

Inhibition of ß-catenin signaling by nongenomic action of orphan nuclear receptor Nur77.

Dysregulation of ß-catenin turnover due to mutations of its regulatory proteins including adenomatous polyposis coli (APC) and p53 is implicated in the pathogenesis of cancer. Thus, intensive effort is being made to search for alternative approaches to reduce abnormally activated ß-catenin in cancer cells. Nur77, an orphan member of the nuclear receptor superfamily, has a role in the growth and apoptosis of cancer cells. Here, we reported that Nur77 could inhibit transcriptional activity of ß-catenin by inducing ß-catenin degradation via proteasomal degradation pathway that is glycogen synthase kinase 3ß and Siah-1 independent. Nur77 induction of ß-catenin degradation required both the N-terminal region of Nur77, which was involved in Nur77 ubiquitination, and the C-terminal region, which was responsible for ß-catenin binding. Nur77/ΔDBD, a Nur77 mutant lacking its DNA-binding domain, resided in the cytoplasm, interacted with ß-catenin, and induced ß-catenin degradation, demonstrating that Nur77-mediated ß-catenin degradation was independent of its DNA binding and transactivation, and might occur in the cytoplasm. In addition, we reported our identification of two digitalis-like compounds (DLCs), H-9 and ATE-i2-b4, which potently induced Nur77 expression and ß-catenin degradation in SW620 colon cancer cells expressing mutant APC protein in vitro and in animals. DLC-induced Nur77 protein was mainly found in the cytoplasm, and inhibition of Nur77 nuclear export by the CRM1-dependent nuclear export inhibitor leptomycin B or Jun N-terminal kinase inhibitor prevented the effect of DLC on inducing ß-catenin degradation. Together, our results demonstrate that ß-catenin can be degraded by cytoplasmic Nur77 through their interaction and identify H-9 and ATE-i2-b4 as potent activators of the Nur77-mediated pathway for ß-catenin degradation.

Polyphenols from Parabarium huaitingii and their positive inotropic and anti-myocardial infarction effects in rats.

Eight phenolic compounds, including (-)-epicatechin (1) and seven proanthocyanidins (2-8), were obtained from the butanol extract of Parabarium huaitingii (PHB). Their chemical structures were identified based on analyses of mass spectra (MS), NMR, CD spectra, and partial acid catalyzed thiolytic degradation. The observation made by laser scanning confocal microscope found a significant increase of the concentration of intracellular Ca²+ ([Ca²+](i)) in single myocytes when the PHB was added, while compounds 1 and 3 had the same physiological effect. Further investigations showed PHB had a dose-dependent positive inotropic effect on isolated right atria and papillary muscle of left ventricle of the rat, while having no significant influence on the spontaneous beating rate of the isolated right atria. The inotropic effect of PHB could be greatly abolished by pretreating the myocardium in Ca²+-free solution. These findings indicated that PHB could significantly increase [Ca²+](i) in myocytes, which was greatly dependent on the influx of extracellular Ca²+. Compounds 1 and 3 might be the effective ingredients of the inotropic effect of PHB. In addition, PHB could also significantly decrease the infarct size of the heart on acute myocardial infarction (AMI) model rats, which suggested its myocardial protective effect on ischemic myocardium. The positive inotropic effect of PHB, together with its myocardial protective effect on AMI, suggested that PHB had a promising potential for the prevention and treatment of heart failure, especially the one that was caused by AMI.

Study of the mechanisms by which Sambucus williamsii HANCE extract exert protective effects against ovariectomy-induced osteoporosis in vivo.

UNLABELLED: The purpose of this study is to investigate the dose-dependent effects of SWH on bone properties and the mechanism involved in mediating the osteoprotective actions of SWH. The results indicated that SWH could improve bone properties by inhibiting the process of bone resorption and stimulating the process of bone formation. INTRODUCTION: Our previous study showed that Sambucus williamsii HANCE (SWH) improved trabecular bone mass and cortical bone strength in ovariectomized (OVX) rats. The purpose of this study is to investigate the dose-dependent effects of SWH on bone properties and the mechanism involved in mediating the osteoprotective actions of SWH. METHODS: Three-month-old C57BL/6J mice were fed a phytoestrogen-free diet and subjected to either ovariectomy or sham operation. OVX mice were treated with genistein (50 mg/kg), or a low (200 mg/kg), medium (500 mg/kg), or high (1,000 mg/kg) dose of SWH extract. RESULTS: SWH could dose-dependently decrease urinary Ca excretion and increase serum Ca level in OVX mice. It could increase tibial bone mineral density and exert beneficial effects on the microarchitecture of trabecular bone in the OVX mice. SWH suppressed the ovariectomy-induced expression of Cbfa1 mRNA and cathepsin K mRNA and enhanced the ratio of OPG/RANKL mRNA expression in the tibia. In vitro study showed that SWH dramatically reduced the number of TRAP-positive cells in RANKL-induced RAW 264.7 cells. CONCLUSIONS: The present study indicated that SWH could improve bone properties by inhibiting the process of bone resorption and stimulating the process of bone formation.

Effects of total flavonoids and flavonol glycosides from Epimedium koreanum Nakai on the proliferation and differentiation of primary osteoblasts.

In a bioassay-guided drug screening for anti-osteoporosis activity, eight flavonol glycosides were isolated from Epimedium koreanum Nakai, which is traditionally widely used in China for the treatment of impotence and osteoporosis. The effects of total flavonoids and flavonol glycosides on the proliferation and differentiation of rat calvarial osteoblast-like cells were evaluated by the MTT method and measuring the activity of alkaline phosphatase (ALP activity). Total flavonoids (1.2 x10(-2) to 6.0 x10(-7) mg/ml) and flavonol glycosides (2.0 x10(-5) to 1.0 x10(-9) mol/l) exhibited a strong inhibition on the proliferation of primary osteoblasts at most concentrations. However, the total flavonoids and icariin significantly promoted the differentiation of primary osteoblasts. The results suggested that flavonoids from E. koreanum Nakai may improve the development of osteoblasts by promoting the ALP activity; and icariin might be one of the active constituents facilitating the differentiation of osteoblasts.

Microbial transformation of curcumol by Cunninghamella blakesleana.

Microbial transformation of curcumol (1) by Cunninghamella blakesleana (AS 3.970) yielded six metabolites. On the basis of spectral data, their structures were elucidated as 14-hydroxy-9E curcumol (2), 10R,14-dihydroxy curcumol (3), 10S,14-epoxy curcumol (4), 10R,14-epoxy curcumol (5), 10S,14-epoxy-7,11-dehydrocurcumol (6), 10R,14-epoxy-7,11-dehydrocurcumol (7), respectively. Among them 2, 3, 6 and 7 are new compounds; 4 and 5, 6 and 7 are two pairs of epimers.

Two new triterpenoid saponins from Ardisia crenata.

Two new triterpenoid saponins, ardisicrenoside K (1) and ardisicrenoside L (2), have been isolated from the roots of Ardisia crenata Sims. Their structures have been determined as 3beta-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl-(1 --> 4)-[beta-D-glucopyranosyl-(1 --> 2)]-alpha-L-arabinopyranosyl]-13beta,28-epoxy-16-oxo-30,30-dimethoxyoleanane and 3beta-O-[beta-D-xylopyranosyl-(1 --> 2)-beta-D-glucopyranosyl-(1 --> 4)-[beta-D-glucopyranosyl-(1 --> 2)]-alpha-L-arabinopyranosyl]-13beta,28-epoxy-16alpha,20-dihydroxyoleanane by means of chemical evidences and spectral analysis. Their weak anti-fungal activity against the plant pathogenic fungus Pyricularia oryzae was evaluated in vitro.

9, 10-Dihydrophenanthrene derivatives from Pholidota yunnanensis and scavenging activity on DPPH free radical.

Three new 9,10-dihydrophenanthrene derivatives named phoyunnanins A-C, together with six known 9,10-dihydrophenanthrene constituents, 4,4',7,7'-tetrahydroxy-2,2'-dimethoxy-9,9',10,10'-tetrahydro-1,1'-biphenanthrene (4), lusianthridin (5), eulophiol (6), 2,4,7-trihydroxy-9,10-dihydrophenanthrene (7) and imbricatin (8), were isolated from the 60% ethanolic extract of air-dried whole plant of Pholidota yunnanensis Rolfe. The structures of phoyunnanins A-C were established as 6-[2'-(3',3''-dihydroxy-5'-methoxybibenzy)]-4,7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (1), 6-[6'-(trans-3',3''-dihydroxy-5'-methoxystilbeny)]-4,7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (2) and 4,4',7,7'-tetrahydroxy-2,2'-dimethoxy-9,9',10,10'-tetrahydro-1,6'-biphenanthrene (3), respectively, on the basis of the spectroscopic analysis. All eight compounds (1-8) were found to show the DPPH free radical scavenging activity with EC(50) from 8.8 to 55.9 microM.

Triterpenoid saponins from Stauntonia chinensis.

A new triterpenoid saponin, named stauntoside A (1) along with four known saponins (2,3,4,5) was isolated from Stauntonia chinensis DC., (Lardizabalaceae). Their structures were elucidated by spectroscopic analysis and chemical methods as 3-O-alpha-L-arabinopyranosyl-30-norhederagenin -28-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (1), 3-O-alpha-L-arabinopyranosyl-30- norhederagenin-28-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (2), 3-O-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl-30-norhederagenin-28-O-alpha-L- rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (3), 3-O-alpha-L- arabinopyranosyl-hederagenin-28-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D- glucopyranosyl ester (4), 3-O-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl-hederagenin -28-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (5). The (1)H and (13)C NMR data for Glycoside L-G1 or Sinofoside A are paradox in the reported before. Thus, the structure elucidation of saponin 2, known as Glycoside L-G1 or Sinofoside A, was discussed and the unambiguous assignments were given.

Three major urinary metabolites of sinomenine in rats.

Urinary metabolites of sinomenine were investigated in rats after intragastric administration. Three major metabolites were obtained and characterised as 4-hydroxy-3,7,7-trimethoxy-17-methyl-(9alpha,13alpha,14alpha)-morphinan-6-one (1), 7,8-didehydro-4-hydroxy-3,7-dimethoxy-17-methyl-N-oxide-(9alpha,13alpha,14alpha)-morphinan-6-one (2), and 7,8-didehydro-4-hydroxy-3,7-dimethoxy-(9alpha,13alpha,14alpha)-morphinan-6-one (3). Their structures have been elucidated on the base of spectral analysis, among which 1 and 2 were new compounds.

Total synthesis of two new dihydrostilbenes from Bulbophyllum odoratissimum.

A total synthetic route of two new dihydrostilbenes 5-(2-benzo[1,3]dioxole-5-ylethyl)-6-methoxy benzo[1,3]dioxole-4-ol (1) and 5-(2-benzo[1,3]dioxole-5-ylethyl)benzo[1,3]dioxole-4,7-diol (2), which were isolated from Bulbophyllum odoratissimum Lindl. with significant cytotoxicity toward human cancer cell lines, was developed via Horner reaction etc. The natural products 1 and 2 were obtained in 10.5% and 3.3% overall yield, respectively.

Down-regulation of prostate specific antigen in LNCaP cells by flavonoids from the pollen of Brassica napus L.

The pollen of Brassica napus L. has been used in China to treat benign prostatic hyperplasia (BPH) for over decades. In this study, the pollen of Brassica napus L. was extracted successively with chloroform, ethyl acetate and ethanol. The ethyl acetate extract showed strong activity in decreasing the secretion of prostate specific antigen (PSA) in LNCaP cells as compared to two other extracts, measured by ELISA with finasteride as positive control in the assay. Five flavonoids were subsequently isolated from the active extract using bioassay-guided fractionation. They were Naringenin (1); Luteolin (2); Kaempferol (3); Kaempferol 3-(3-E-p-coumaroyl-alpha-L-rhamnopyranoside) (4); and Kaempferol 3-(2,3-di-E-p-coumaroyl-alpha-L-rhamnopyranoside) (5). All these compounds inhibited PSA secretion significantly, with IC50 values in the range of 5-50 microM. Compounds 2, 4 and 5 showed moderate cytotoxicity to LNCaP cells within the active concentration range, while compounds 1 and 3 showed no cytotoxicity. Further studies on the mechanism action of these compounds were performed by evaluating their activation of estrogen receptor (ER) and antagonistic activities on androgen receptor (AR) in cell-based reporter gene assays. All compounds described here were first isolated from the pollen of Brassica napus L.

Waveplate analyzer using binary magneto-optic rotators.

We demonstrate a simple waveplate analyzer to characterize linear retarders using magneto-optic (MO) polarization rotators. The all-solid state device can provide highly accurate measurements for both the retardation of the waveplate and the orientation of optical axes simultaneously.

Studies on the chemical constituents of Valeriana fauriei Briq.

Nine compounds were isolated from the roots of Valeriana fauriei Briq, of which one is a new germacrane-type sesquiterpenoid named as valerianin E and its structure was elucidated as bicyclo[8, 1, 0] 5beta-hydroxyl-7beta-acetoxyl-5alpha,11, 11'-trimethyl-E-1(10)-ene-4alpha, 15-olide (1). In addition, two were first reported from this genus and the others were isolated for the first time from the title plant.

Inhibitory effects of eicosapentaenoic acid (EPA) on the hypoxia/reoxygenation-induced tyrosine kinase activation in cultured human umbilical vein endothelial cells.

We have previously reported that the n-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) inhibited the abnormal gap junctional intercellular communication (GJIC) induced by hypoxia/reoxygenation (H/R) via suppressing tyrosine kinase (TK) activation (Zhang et al., Prostaglandins Leukot Essent Fatty Acids, 1999; 61: 33-40). However, the mechanisms by which EPA-inhibited TK activation remained unidentified. In this study we investigated whether reactive oxygen species (ROS) and growth factor-receptor systems would contribute to the H/R-induced TK activation or not. The results showed that H/R-induced ROS production, which reached the peak after 30 min of reoxygenation. Pretreatment with 10 microM EPA significantly inhibited this ROS production. However, the TK inhibitor genistein (10 microM) failed to inhibit the generation of ROS, although it completely inhibited TK activation. On the other hand, the ROS inhibitor DMSO (0.5% v/v) showed little effect on TK activation while it significantly blocked ROS production. Further EPA and genistein, but not DMSO and superoxide dismutase (SOD, 300 U/ml), prevented cells from GJIC injury induced by H/R. Moreover, EPA protected against VEGF-induced reduction in GJIC and phosphorylation of connexin 43. These data suggest that growth factor, but not ROS, might be involved in the EPA-inhibited TK activation induced by H/R.

Bioactive saponins from Dioscorea futschauensis.

A new anti-neoplastic spirostanol saponin, (25S)-spirost-5-en-3 beta, 27-diol-3O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl (1-->3)]-beta-D-glucopyranoside and three known compounds viz. prosapogenin A of dioscin, dioscin and gracilin were isolated from Dioscorea futschauensis by bioactivity-guided fractionation. Their structures were elucidated mainly by means of spectroscopic analysis. Their bioactivity against Pyricularia oryzae and cytotoxic activity on ts-FT210 cell line was evaluated.

The cytotoxicity of methyl protoneodioscin (NSC-698791) against human cancer cell lines in vitro.

Methyl protoneodioscin (NSC-698791) is a furostanol saponin isolated from the rhizome of Dioscorea collettii var. hypoglauca (Dioscoreaceae), a Chinese herbal remedy for the treatment of cervical carcinoma, carcinoma of the urinary bladder and renal tumor for centuries. In order to systematically evaluate its potential anticancer activity, methyl protoneodioscin cytotoxicity was tested in vitro against 60 human cancer cell lines in the NCI's (National Cancer Institute, USA) anticancer drug screen. As a result, methyl protoneodioscin was cytotoxic against all the test cell lines from leukemia and solid tumors in the NCI's human cancer panel, especially selectively against one non-small cell lung cancer (NSCLC) line (A549/ATCC), one colon cancer line (HCT-116), two central nenous system (CNS) cancer lines (SF-539 and SNB-75), one melanoma line (M14), one renal cancer line (CAKI-1), one prostate cancer (DU-145) and two breast cancer lines (HS 578T and MDA-MB-435) with GI50 < or = 2.0 microM. The selectivity between these nine most sensitive lines and the least sensitive line (TK-10) was from 22- to 30- fold. In the same cancer subpanel, a selectivity at GI50 level of more than 15-fold was observed between A549/ATCC and EKVX (NSCLC), between CAKI-1 and TK-10, A498 (renal cancer), respectively. In general the CNS cancer was the most sensitive subpanel, while renal cancer was the least sensitive subpanel. Based on an analysis of COMPARE computer program with methyl protoneodioscin as a seed compound, no compounds in the NCIs anticancer drug screen database have similar cytotoxicity patterns (mean graphs) to that of methyl protoneodioscin, indicating a potential novel mechanism of anticancer action involved.