Seven compounds were isolated from Onychium japonicum by macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified by NMR, MS and other spectroscopic methods as onychone A (1), quercetin (2), quercetin-3-O-α-L-rhamnoside (3), kaempferol-7-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnopyranoside (5), (-)-prunin (6), and norathyriol (7). Compound 1 is a novel macrocyclic flavonoid, and all the others are reported from this plant for the first time. In vitro cytotoxic activities of compounds 1-7 were evaluated by MTS testing with five cancer cell lines. Compound 7 exhibited weak cytotoxicity against tumor cell lines A549, SMMC-7721, and SW480.

This study aimed to construct the dual expression vectors of wide type or N187D mutant P2X7 receptor and intracellular domain of Notch1 (ICN1) linked by 2A peptide to coexpress them in leukemia cells so as to lay a foundation for further investigating the role of P2X7 in development of leukemia. Overlap PCR was used to construct the dual expression vectors encoding wide type or N187D mutant type P2X7 receptor and ICN1 linked by the self-cleaving 2A sequence. The results showed that stable expressing cell lines were obtained by retroviral infection followed by cell sorting after DNA sequence analysis. RT-PCR, Western blot, intracellular free calcium concentration analysis were used to verify the functionally successful construction of K562 cell line expressing P2X7 receptor alone or with ICN1. DNA sequence analysis revealed that all construction were right. The infection efficiency of packaged constructed virus ranged from 40% to 70% for K562 cells. Stable infected cell line was obtained by cell sorting. RT-PCR analysis revealed that P2X7 receptor and/or ICN1 could be detected at high level in their stable infected cell lines, respectively. Western blot analysis also showed that P2X7 receptor was highly expressed in cell line infected by virus with P2X7 receptor. Sustained increase in intracellular free calcium concentration ([Ca(2+)]i) could be observed in K562 cells overexpressing either type of P2X7 receptor upon stimulation with BzATP. It is concluded that the wide type or N187D mutant P2X7 receptor and ICN1 are simultaneously and functionally over-express in leukemia cells, which lay a foundation for further studying the role of P2X7 receptor in the development of leukemia.
Gene Expression ; Genetic Vectors ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; Receptor, Notch1 ; genetics ; Receptors, Purinergic P2X7 ; genetics ; Retroviridae ; genetics

OBJECTIVETo evaluate the urodynamic characteristics of the chronic impairment of cauda equina caused by lumbar disk herniation.

METHODSClinical data and urodynamic parameters of 67 male patients with lumbar disk herniation were retrospectively analyzed. Lower urinary obstruction was excluded from the cohort using the Lin-PURR analysis. Patients were divided into group A (normal detrusor function), group B (detrusor underactivity) and group C (detrusor areflexia) according to the detrusor contraction function analyzed in Lin-PURR. Clinical data and urodynamic parameters were analyzed statistically between these groups.

RESULTSThe category of the detrusor contraction function had a significant effect on the urodynamic parameters. There were significant differences in the maximum flow rate (Q(max)), maximum pressure (P(max)), pressure at the maximum flow (P(det Qmax)) and post-voiding residual urine (PVR) among group A, B and C. There were significant differences in the first sensation volume of the bladder and the maximum cystometric capacity between group A and C, B and C, but no significance was found between group A and B. There was no significant difference in age, disease duration, and compliance of the bladder among 3 groups.

CONCLUSIONSUrodynamic study is important in exploring the severity of the chronic impairment of cauda equina caused by lumbar disk herniation. Detrusor areflexia and loss of bladder sensory indicate more severe degree of impairment of the cauda equine. Q(max) and PVR are helpful in early diagnosis of the chronic impairment of cauda equina.

Adult ; Aged ; Aged, 80 and over ; Cauda Equina ; Chronic Disease ; Humans ; Intervertebral Disc Displacement ; complications ; physiopathology ; Male ; Middle Aged ; Nerve Compression Syndromes ; etiology ; physiopathology ; Retrospective Studies ; Urodynamics

Adult ; Bone Neoplasms ; diagnosis ; Cervical Vertebrae ; pathology ; Humans ; Lipoma ; diagnosis ; Male

Objective To explore the risk factors of patent ductus arteriosus (PDA) patients with thrombocytopenia after PDA interventional occlusion.Methods Thrombocytopenia occurred in 14 out of 350 patients underwent PDA occlusion.Age,gender,body weight,PDA size,occluder size,mean pulmonary arterial pressure,the dose of heparin,the manufacturer of occluder,residual shunt after operation were analyzed.The recovery time of different grades of thrombocytopenia was observed.Results Multiwariate logistic regression showed that the PDA size (OR =2.238,P ＜ O.05),the dose of heparin (OR =3.247,P ＜ 0.05),residual shunt after operation (OR =1.912,P ＜ 0.01) were the independent risk factors of thrombocytopenia after PDA occlusion.The recovery time of mild thrombocytopenia was (7 ± 2)days without treatment.The recovery time of moderate thrombocytopenia was (12 ± 4) days with glucocorticoids treatment.The recovery time of severe thrombocytopenia was (21 ± 7) days with platelet transfusion.Conclusions The occluder size,dose of heparin,residual shunt are the independent risk factors of thrombocytopenia after PDA interventional occlusion.Recover time of thrombocytopenia after PDA interventional occlusion is closely related to the severity of thrombocytopenia.

OBJECTIVETo evaluate the application of multi-detector row CT (MDCT) and CT angiography (CTA) for detecting early signs of acute bowel ischemia (ABI) in experimental porcine models.

METHODSTwelve pigs were assigned to four groups with 3 in each group. The digital subtraction angiography of superior mesenteric artery (SMA) and the embolization of branches of SMA with gelatin sponge and blood clot were performed by percutaneous transfemoral artery puncture and catheterization. MDCT pre- and post-contrast scanning in the arterial, venous and delay phase and CTA with three-dimensional reconstruction were carried out at pre-operation, 3 h, 6 h, 9 h, and 12 h after occlusion. The normal mesenteric vascular anatomy, arterial occlusion, mesentery and bowel changes, and dynamic change were evaluated.

RESULTSABI changes were identified pathologically in all the 12 experimental pigs, and the severity of ischemia increased over time after embolization. CTA showed all 57 embolized branches of SMA and 29 of 34 unoccluded arterial branches with 5 false-positive vessel occlusions. The sensitivity and specificity of CTA were 100% and 85.3%, respectively. Thin-slab maximum intensity projection (TSMIP) revealed the disappearance of distal comb-like vessel branches and brush-like vasa recta, which were clearly delineated in the normal bowel segments. Using this criterion, TSMIP correctly defined 23 of 24 ischemic bowel segments and all the 12 normal bowel segments with a sensitivity of 95.8% and a specificity of 100%.

CONCLUSIONSMDCT and CTA reliably define normal and occluded mesenteric vessels in the pig. It can easily detect ischemic bowel segment by identified early changes of ischemia. The early direct ischemic signs are occluded vessels, the disappearance of distal comb-like branches or brush-like vasa recta, and poor bowel enhancement. The early indirect sign is bowel dilatation with fluid collection.

Angiography ; methods ; Animals ; Female ; Intestinal Diseases ; diagnostic imaging ; etiology ; Ischemia ; diagnostic imaging ; etiology ; Mesenteric Arteries ; diagnostic imaging ; Mesenteric Vascular Occlusion ; complications ; diagnostic imaging ; Mesentery ; blood supply ; Swine ; Tomography, X-Ray Computed ; methods

OBJECTIVETo explore the patterns of innervation of cervical facet joints and determine the pathways from facet joints to dorsal root ganglions (DRGs) in order to clarify the causes of diffuse neck pain, headache, and shoulder pain.

METHODSForty-two male-Sprague-Dawley rats, weighing 250-300 g, were randomly divided into three groups: Group A (n=18), Group B (n=18), and Group C (n=6). Under anesthesia with intraperitoneal pentobarbital sodium (45 mg/kg body weight), a midline dorsal longitudinal incision was made over the cervical spine to expose the left cervical facet joint capsule of all the rats under a microscope. The rats in Group A underwent sympathectomy, but the rats in Group B and Group C did not undergo sympathectomy. Then 0.6 microlitre 5% bisbenzimide (Bb) were injected into the C1-2, C3-4 and C5-6 facet joints of 6 rats respectively in Group A and Group B. The holes were immediately sealed with mineral wax to prevent leakage of Bb and the fascia and skin were closed. But in Group C, 0.9% normal saline was injected into the corresponding joint capsules. Then under deep re-anesthesia with intraperitoneal pentobarbital sodium (45 mg/kg body weight), C1-C8 left DRGs in all rats and the sympathetic ganglions in Group B were obtained and the number of the labeled neurons was determined.

RESULTSNeurons labeled with Bb were present in C1-C8 DRGs in both Group A and Group B, and sympathetic ganglions in Group B. In the C1-2 and C3-4 subgroups, labeled neurons were present from C1 to C8 DRGs, while in C5-6 subgroups they were from C3 to C8. The number of Bb(+) neurons after sympathectomy was not significantly different in the injected level from that without sympathectomy. But in the other levels, the number of Bb(+) neurons after sympathectomy was significantly less than that without sympathectomy.

CONCLUSIONSThe innervation of the cervical facet joints is derived from both sensory and sympathetic nervous system, and DRGs are associated with sympathetic ganglions through nerve fibers outside the central nerve system.

Animals ; Cervical Vertebrae ; innervation ; Ganglia, Spinal ; cytology ; Ganglia, Sympathetic ; cytology ; Male ; Neurons, Afferent ; cytology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDExposure of cells to sublethal concentrations of hydrogen peroxide (H2O2) can alleviate subsequent oxidative stress-induced apoptosis. We assessed the effects of H2O2 preconditioning on the therapeutic potential of human umbilical cord Wharton's Jelly mesenchymal stem cells (WJ-MSCs) in a murine model of myocardial infarction.

METHODSWJ-MSCs were incubated in the media for 2 hours with or without 200 µmol/L H2O2. Mice underwent left anterior descending coronary artery ligation, and received injection of phosphate buffered saline, 1×10(6) WJ-MSCs, or 1×10(6) H2O2 preconditioned WJ-MSCs 3 hours later via tail vein. Echocardiography was performed 0, 7, 14 and 28 days after surgery, and the mice were euthanized on day 28 for histological analysis. In vitro cytokine concentrations in the WJ-MSC cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). The effect of WJ-MSC cell supernatant on the migration and proliferation of endothelial cells were observed by transwell migration and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide (MTT) assays.

RESULTSEchocardiographic measurements revealed a significant improvement in the left ventricular contractility of the WJ-MSCs-H2O2 group compared to the WJ-MSCs group. Histological analysis revealed increased neovascularization and reduced myocardial fibrosis in the WJ-MSCs-H2O2 group compared to the WJ-MSCs group. Pretreatment of WJ-MSCs with H2O2 increased the secretion of interleukin-6 (IL-6) into the cell culture supernatant by approximately 25-fold. The culture supernatant from WJ-MSCs-H2O2 significantly increased the migration and proliferation of endothelial cells; these effects could be blocked using an anti-IL-6 antibody.

CONCLUSIONSThis study demonstrates that H2O2 preconditioning significantly enhanced the therapeutic potential of WJ-MSCs, possibly by stimulating the production of IL-6 by WJ-MSCs, which may cause migration and proliferation of endothelial cells and increase neovascularization.

Animals ; Cell Movement ; physiology ; Echocardiography ; Enzyme-Linked Immunosorbent Assay ; Humans ; Hydrogen Peroxide ; pharmacology ; Immunohistochemistry ; Interleukin-6 ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Myocardial Infarction ; pathology ; therapy ; Reactive Oxygen Species ; metabolism ; Wharton Jelly ; cytology

Breast cancer is one of the leading causes of cancer death worldwide. This study aimed to analyze the expression of centromere protein H (CENP-H) in breast cancer and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells, immortalized mammary epithelial cell lines, and breast cancer cell lines, we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells. We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry, and detected high CENP-H expression in 134 (43.6%) samples. Statistical analysis showed that CENP-H expression was related with clinical stage (P = 0.001), T classification (P = 0.032), N classification (P = 0.018), and Ki-67 (P < 0.001). Patients with high CENP-H expression had short overall survival. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival. Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.
Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Breast ; cytology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Rate ; Up-Regulation