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

OBJECTIVETo investigate over-expression of wild-type alpha-synuclein inducing the aberrant aggregation of alpha-synuclein in HEK293 cell in vitro.

METHODSThe cDNA encoding the human alpha-synuclein without the stop code was cloned into PGEM T-easy vector. Using enzyme map and DNA sequencing analyzed and determined the recombinant plasmid, and then sub-clone the alpha-synuclein cDNA fragment into pEGFP-N1 vector. The recombinant plasmids alpha-synuclein-pEGFP were transfected into HEK293 cells by lipofectamin 2000. The aberrant aggregation of alpha-synuclein was measured by EGFP fluorescence, anti-alpha-synuclein immunocytochemistry. The inclusions in the cultured cells were identified with HE staining.

RESULTSThe restriction enzyme map suggested that eukaryotic expression vector for human wild-type alpha-synuclein gene was constructed successfully. By EGFP fluorescence, anti-alpha-synuclein immunocytochemistry, it could be observed that the alpha-synuclein protein could aggregate in cytoplasm and the Lewy body-like inclusions found in cytoplasm of cultured cells.

CONCLUSIONThe over-expression of wild-type alpha-synuclein can induce protein aberrant aggregation and Lewy body-like inclusions formation in cytoplasm of HEK293 cell in vitro.

Cells, Cultured ; Gene Expression ; Humans ; Immunohistochemistry ; Inclusion Bodies ; metabolism ; Lewy Bodies ; metabolism ; Parkinson Disease ; genetics ; metabolism ; alpha-Synuclein ; genetics ; metabolism

OBJECTIVETo study the effects of high frequency oscillatory ventilation (HFOV) combined with incremental positive end-expiratory pressure (IP) on respiratory and circulatory functions, and lung histopathology of dogs with smoke inhalation injury.

METHODSAfter being treated with conventional mechanical ventilation, 12 dogs were inflicted with severe smoke inhalation injury and divided into group HFOV and group HFOV+IP according to the random number table, with 6 dogs in each group. Then they received corresponding ventilation for 8 hours respectively. Blood gas analysis results (pH value, PaO2 and PaCO2 levels) and hemodynamic parameters [heart rate, mean arterial pressure (MAP), pulmonary arterial pressure (PAP), central venous pressure (CVP), cardiac output (CO)] were recorded before injury, immediately after injury, and at post ventilation hour (PVH) 2, 4, 6, and 8. The dogs of two groups were sacrificed at PVH 8. A healthy dog without any treatment and a dog with smoke inhalation injury but no subsequent treatment were sacrificed in addition. Lung tissues of all dogs were obtained for histopathological observation. Lung injury score examination was conducted in both groups. Data were processed with rank sum test, analysis of variance of repeated measurement, and LSD- t test.

RESULTS(1) The PaO2 levels in both groups were significantly decreased immediately after injury, compared with those before injury (with t values respectively 4.960, 5.310, P values all below 0.01). The PaO2 levels in both groups from PVH 2 to PVH 8 were significantly increased, compared with those observed immediately after injury (with t values from 4.930 to 6.050, P values all below 0.01). At PVH 2, 4, and 8, PaO2 levels in group HFOV+IP were significantly higher than those in group HFOV (with t values from 3.775 to 5.774, P values all below 0.01); no statistically significant differences were observed in pH value and PaCO2 level at each time point between two groups (with t values from 0.002 to 0.997, P values all above 0.05). (2) There were no statistically significant differences in MAP, PAP, and CVP within two groups at each time point (with F values from 1.316 to 4.959, P values all above 0.05). In group HFOV, heart rate from PVH 2 to PVH 8 was significantly lower than that observed immediately after injury (with t values from 3.780 to 8.970, P values all below 0.01). In group HFOV+IP, CO at PVH 4, 6, and 8 was significantly lower than that observed immediately after injury (with t values from 3.990 to 11.200, P values all below 0.01). There were no statistically significant differences in MAP, PAP, and CVP between two groups at the same time point (with t values from 0.089 to 2.123, P values all above 0.05). At PVH 4, 6, and 8, heart rate in group HFOV+IP was higher than that in group HFOV (with t values from 2.931 to 7.229, P < 0.05 or P < 0.01), while CO was lower (with t values from 4.297 to 11.206, P values all below 0.01). (3) Compared with those of the healthy dog, inflammatory cell infiltration and bleeding in the lung were observed in alveolar space in both group HFOV and group HFOV+IP, while the degree was less serious than that of the dog with smoke inhalation injury only. Compared with those of group HFOV, inflammatory cell infiltration in group HFOV+IP was less significant, the alveolar structure was relatively intact, and no thickening of alveolar walls was observed. The lung injury score in group HFOV [(3.27 ± 0.24) points] was higher than that of group HFOV+IP [(2.79 ± 0.31) points, t = 27, P < 0.05].

CONCLUSIONSHFOV combined with IP can improve gas exchange and alleviate pulmonary injury without any adverse effect on blood gas analysis or hemodynamic parameters. Therefore, it may be considered as an appropriate mode of ventilation for the treatment of smoke inhalation injury.

Animals ; Blood Gas Analysis ; Burns, Inhalation ; physiopathology ; therapy ; Disease Models, Animal ; Dogs ; Hemodynamics ; High-Frequency Ventilation ; Male ; Positive-Pressure Respiration ; Smoke ; adverse effects