Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
Amphibian Venoms ; chemistry ; Animals ; Bufanolides ; analysis ; chemistry ; Bufo bufo ; Chromatography, High Pressure Liquid ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

BACKGROUND: According to the current most prevailed posterior pedicle screw reduction and Cage fusion for spondylolisthesis, the previous focus is to restore the vertebral sagittal displacement, and whether restoring disc height is needed and the degree of recovery remain unclear. Moreover, there is still a lack of better quantitative indicators to restore the disc height.OBEJCTIVE: By comparing the surgery of lifting reduction, to study the clinical effect of distraction and lifting reduction combined with posterior intervertebral fusion in the treatment of lumbar spondylolisthesis. METHODS: Sixty patients with L5 spondylolisthesis located in L5/S1 were selected, and were divided into two groups based on the surgery methods (n=30 per group): group A (single lifting reduction group) and group B (lifting and distracting reduction group). The patients in the group B were given distracting reduction amid insertion of 12 mm-height Cage; while the patients in the group A were subjected to lifting reduction, and intervertebral fusion with 8 mm-height Cage. The general information and the Visual Analogue Scale, Japanese Orthopaedic Association and Oswestry Disability Index scores at baseline, postoperative 1 month and last follow-up, as well as the postoperative correction and bone fusion were recorded. Additionally, the clinical effectiveness was assessed by Macnab score. RESULTS AND CONCLUSION: (1) At postoperative 1 month and last follow-up, the disc height, disc height/vertebral height, neuroforamen height and clinical effectiveness in the group B were superior to those in the group A (P < 0.05). (2) To conclude, the pedicle screw system combined with posterior intervertebral cage fusion for spondylolisthesis, can rapidly restore the disc height, contribute to restructure better biomechanics of spine. Furthermore, it significantly improves clinical systems, is easy to operate, and achieves better efficacy; therefore, it is recommended firstly.

Adenoids ; diagnostic imaging ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Hypertrophy ; Magnetic Resonance Imaging ; Male ; Radiographic Image Enhancement ; methods ; Tomography, Spiral Computed ; methods

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.
Animals ; Bufanolides ; chemistry ; isolation & purification ; Bufo bufo ; Injections ; Skin ; chemistry

Objective To investigate the possible role of interstitial cell of Cajal (ICE)in the pathogenesis of diabetic gastroparesis and the protective effect of"Xiaopi prescription (消痞方)".Methods Fifty healthy male Spregue-Dawley (SD) rats were randomly divided into 5 groups (each n=10):normal, model,and 3 Xiaopi prescription groups:low,middle and high dosages.Diabetes was induced by intravenous injection of alloxan,and equal amount of normal saline was intravenously injected in the normal group.Gastric lavage method was used to administer the traditional Chinese medicine decoction of Xiaopi prescription in corresponding amount (5,10 and 20 g?kg~(-1)?d~(-1)) in respective low,middle and high dosage groups.In the normal control group and diabetic model group,only equal amount of normal saline was administered into the stomach.Gastric emptying rate was measured by method of nutritious semisolid paste;c-kit positive cells of ICC were quantitatively measured with immunohistochemistry assay and computer image analysis system;c-kit mRNA positive cells were quantitatively measured with in situ hybridization and computer image analysis system.Results①Gastric emptying rate:The rate was significantly lower in the model group than that in the normal control group (P0.05),but higher than those in the model group and the low dosage group (all P0.05).②c-kit immmunohistochemistry:c-kit positive cell presented yellow in color,and its membrane was stained yellow,this kind of cells primarily were distributed around the neural plexus in the inter-space between the circular and longitudinal muscular fibers, and around the ganglionic cells forming"sheath-like"structure.The results of numbers of c-kit positive cells in the various groups:the number of the cells in the model group was significantly lower than that in the normal group (P0.05),but the numbers in the former two dosage groups were obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal group,middle and high dosage groups (all P0.05),but obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal,middle and high dosage groups (all P

Flavonol synthase (FLS) is one of the key enzymes in flavonoids metabolic pathways. In this study, middle sequence was obtained from Carthamus tinctorius transcriptome sequencing results. Full-length cDNAs of FLS was cloned from petals of C. tinctorius to FLS by using RT-PCR and RACE technology. Its full-length cDNA was 1,201 bp, with an open reading frame of 1,101 bp and 336 encoded amino acids. The phylogenetic analysis showed that, FLS gene encoded amino acids in C. tinctorius were highly homologous with amino acids in congeneric Compositae species, especially Rudbeckia laciniata. The pBASTA-FLS plant expression vector was successfully built by the molecular biology method, which lays a foundation for further studying biology functions of the gene and biosynthesis mechanism of flavonoids.
Amino Acid Sequence ; Base Sequence ; Carthamus tinctorius ; classification ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; genetics ; metabolism

OBJECTIVE To identify the valid targets and new drugs of ulcerative colitis (UC), a recurrent and intractable inflammatory bowel disease. METHODS and RESULTS In an in vivo mouse model of DSS-induced colitis, HLJ2 decreased weight loss, colon contracture, disease activity index (DAI), colon mucosa damage index (CMDI) and histopathological index (HI). HLJ2 also decreased myelo?peroxidase(MPO) activity and reduced production of the inflammatory cytokines TNF- α, IL- 1β, andIL- 6. HLJ2 improved intestinal mucosa damage induced by dextran sodium sulfate (DSS) and increased the expression of ZO-1 and claudin-1. Fecal 16s rRNA high-throughput sequencing demon?strated a significant improvement in UC intestinal dysbacteriosis in mice treated with HLJ2, including increased abundance of probiotics such as Lachnospiraceae, Prevotellaceae, and Lactobacillaceae. At the same time there was a reduction in the abundance of pathogenic or conditional pathogenic microor?ganisms such as Bacteroidaceae, Porphyromonadaceae, Deferribacteraceae, and Pseudomonadaceae in HLJ2- treated mice compared with untreated mice. CONCLUSION Our results demonstrated that the XBP1 agonist HLJ2 inhibits inflammation, regulates the intestinal flora, and protects the intestinal mucosa. It is thus a potential therapeutic agent for ulcerative colitis.

Objective To investigate the changes of plasma levels of lysophosphatidic acid (LPA) in the patients with acute cerebral infarction and to observe whether the treatment with Tongxinluo capsules would reduce the plasma LPA levels of the patients with acute cerebral infarction. Methods A total of 70 patients were randomly divided into treatment group and control group. Patients in treatment group were treated with normal method, aspirin (100 mg once a day) and Tongxinluo capsules (4 pills for 3 times per day for 60 d). Patients in control group were treated with only normal method and aspirin (100 mg once a day). Plasma LPA levels were measured consecutively in all patients before treatment, 30 d and 60 d after treatment. The effects of Tongxinluo capsules on plasma LPA levels were observed, and the recurrence rate of cerebral infarction was recorded within 2 months. Results The average levels of plasma LPA in the 2 groups were decreased obviously after the treatments (P<0.05), and the changes of plasma LPA levels in treatment group were significant after treatment for 30 d compared with those in control group (P<0.05). The recurrence rate of cerebral infarction in treatment group was lower than that in control group. Conclusions The plasma LPA levels in the patients with acute cerebral infarction can be decreased significantly after treatment with Tongxinluo capsules. Combined treatment of Tongxinluo capsules with aspirin may reduce the plasma LPA levels and the frequency of cerebral infarction events better.

AIMThe mechanism of vascular endothelial growth factor165 (VEGF165) on intracellular free magnesium ([Mg2+]i) in human umbilical vein endothelial cells (HUVECs) was investigated.

METHODS[Mg2+]i in HUVECs loaded with fluorescent magnesium indicator mag-fura-2 were quantitatively detected the use of intracellular cation measurement system.

RESULTSVEGF165 significantly increased [Mg2+]i in the extracellular Mg2+ and this effect could be blocked by pretreatment with tyrosine kinase inhibitors (tyrphostin A23 and genistein), phosphatidylinositol 3-kinase (PI3K) inhibitors (wortmannin and LY294002) and phospholipase Cgamma (PLCgamma) inhibitor (U73122). In contrast, phospholipase Cgamma (PLCgamma) inhibitor analog (U73343), mitogen-activated protein kinase inhibitors (SB202190 and PD98059) had no effect on the VEGF165-induced [Mg2+]i increase.

CONCLUSIONThe increase of [Mg2+]i by VEGF165 originates from intracellular Mg2+ pool through tyrosine kinase/ PI3K/PLCgamma-dependent signaling pathways.

Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Magnesium ; metabolism ; Neovascularization, Physiologic ; drug effects ; Phosphatidylinositol 3-Kinases ; metabolism ; Phospholipase C gamma ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Vascular Endothelial Growth Factor A ; physiology

To prevent acute graft-versus-host disease (aGVHD), glucosidorum tripterygii tororum (GTT) was used in the murine model. The lethally irradiated C57BL/6 recipients were injected with bone marrow and lymphocyte grafts from BALB/c donors and were treated intraperitoneally with GTT, cyclosporine A (CsA), or methotrexate (MTX). T lymphocytes, adhesion molecules and cytokines were detected by immunohistochemical method, flow cytometry, ELISA and RT-PCR, respectively. The results showed that all the control recipient mice (21/21) died of aGVHD within 30 days, but many recipient mice treated with GTT (19/21), CsA + MTX (13/21) and GTT + CsA (17/21) survived beyond 30 days without obvious signs of aGVHD. The numbers of CD3(+), CD4(+), CD8(+), CD11a(+), CD18(+) lymphocytes in skin and lung decreased markedly by GTT, GTT + CsA and CsA + MTX treatments. The numbers of CD3(+), CD4(+), CD8(+), CD4(+)CD11a(+), CD4(+)CD18(+), CD8(+)CD11a(+), CD8(+)CD18(+) lymphocytes in spleen decreased markedly by GTT, GTT + CsA and CsA + MTX treatments. and the changes of CD3(+), CD4(+), CD8(+) cells in small intestine were not remarkable (P > 0.05) by above mentioned GTT, GTT + CsA and CsA + MTX treatments. The serum concentrations and mRNA expressions of IL-2 and TNFalpha in spleens decreased significantly (P < 0.05); the concentration of IL-10 increased significantly (P < 0.05), the change of IL-4 was not remarkable (P > 0.05) by GTT treatment. It is concluded that the GTT may retain the graft-versus-leukemia (GVL) effect of transplant without aGVHD. The role of GTT in prevention of murine aGVHD is mediated by reduction of T lymphocytes and their subgroups, expression of adhesion molecule, and regulation of cytokine secretion.
Animals ; Bone Marrow Transplantation ; adverse effects ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Female ; Glucosides ; isolation & purification ; therapeutic use ; Graft vs Host Disease ; prevention & control ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Phytotherapy ; Tripterygium ; chemistry