In order to affirm the cardioactive components in Fuzi, we identified a group of aminoalcohol- diterpenoid alkaloids in Fuzi using ultra high-performance liquid chromatography coupled with electrospray ionization mass spectrometer (UPLC-ESI-MS) method. Among a total of forty-one isolated ingredients, thirteen major aminoalcohol-diterpenoid alkaloids were identified by comparing their retention times and MS spectra with those of the reference substances. Moreover, Fuzi samples from different places of origin and with different processing methods were examined and their components displayed a pattern of high similarity, though the relative abundance varies probably due to their different processing methods. Furthermore, the cardiac effect of each identified alkaloid was individually evaluated using the isolated bullfrog heart perfusion experiment. Among the thirteen aminoalcohol diterpenoid alkaloids tested, six of them significantly enhanced the amplitude rates. Taken together, we affirm that the cardioactive components in Fuzi are aminoalcohol-diterpenoid alkaloids, shedding light on future studies of the mechanisms and development of these cardioactive compounds.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Amino Alcohols ; chemistry ; Animals ; Cardiotonic Agents ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Heart ; drug effects ; In Vitro Techniques ; Plant Extracts ; chemistry ; Rana catesbeiana ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo analyze the comorbidities in patients with cerebral palsy (CP) from two perspectives as neurologic subtype and gross motor functions, and find their correlations.

METHODSChildren with cerebral palsy treated in the rehabilitation center from January 2007 to June 2009 received the following examinations: intelligence capacity test, ophthalmologic consultation, language-speech test, brainstem auditory evoked potential and electroencephalogram. They were stratified according to both neurologic subtype and gross motor functions to detect the occurrence of comorbidities.

RESULTSOf all the 354 cases, 166 (46.89%) had mental retardation, 15 (4.24%) auditory limitations, 138 (38.98%) visual disorder, 216 (61.02%) language-speech disorder and 82 (23.16%) epilepsy. The frequency of individual comorbidities were distributed disproportionately between the different neurologic subtypes. Correlation analysis showed that there was a significant correlation between the spastic diplegia and the visual disorder (correlation coefficient = 0.26), between spastic hemiplegia and epilepsy (correlation coefficient = 0.17), between spastic quadriplegia and epilepsy and mental retardation (the correlation coefficient was 0.38 and 0.11, respectively) and between both dyskinetic and mixed children and language-speech disorder (the correlation coefficient was 0.24 and 0.27, respectively). The frequency of individual comorbidities was distributed disproportionately between the different neurologic subtypes and between the different GMFCS levels (P < 0.05), except for the frequency of visual disorders (chi(2) = 1.90, P > 0.05); and with the increase of the GMFCS levels, the burden of the comorbidities were more heavy and the incidence of the comorbidities was higher. Multi-comorbidities were relatively infrequently encountered in those with spastic hemiplegic or spastic diplegic children or patients whose GMFCS levels were I-III, while these entities occurred at a frequent level for those with spastic quadriplegic, dyskinetic, or mixed or children whose GMFCS levels were IV and V, and the differences were significant (P < 0.05). The mean GMFCS levels of children with spastic quadriplegic, dyskinetic or mixed CP were higher than level III, most of them had no ability of ambulation;while the mean GMFCS levels of spastic hemiplegic or spastic diplegic children were below level III, most of them could walk independently.

CONCLUSIONSThere are correlations between the occurrence of the comorbidities such as mental retardation, auditory or visual impairments, language-speech disorders, epilepsy and the cerebral palsy subtype and the gross motor function levels. Clinicians should have a full recognition of these comorbidities, and we should have a cooperation between the different subjects to have an overall evaluation and rehabilitation and to improve the prognosis.

Adolescent ; Cerebral Palsy ; classification ; epidemiology ; Child ; Child, Preschool ; Comorbidity ; Epilepsy ; classification ; epidemiology ; Female ; Humans ; Infant ; Male ; Motor Skills ; classification ; Motor Skills Disorders ; classification ; epidemiology ; Quadriplegia ; classification ; epidemiology ; Vision Disorders ; classification ; epidemiology

Objective To observe the anti-inflammation effects of Chanqin granules on airway neurogenic inflammation induced by fine particulate matter (PM2.5)and mediated by nerve growth factor (NGF)and to explore the therapeutic mechanism. Methods Forty SD rats were randomly divided into 5 groups,namely saline control group, PM2.5 exposure group, anti-NGF group, Chanqin granules group and Huifeining group (Pseudoephedrine Hydrochloride,Chlorpheniramine Maleate and Dextromethorphan Hydrobromide Solution),8 rats in each group. PM2.5 exposure rat model was established by intratracheal drop infusion of PM2.5 suspension. The rats in PM2.5 exposure group were given gastric gavage of normal saline after PM2.5 exposure , and the anti-NGF group was given gastric gavage of saline after PM2.5 exposure and intraperitoneal injection of anti-NGF agent. Chanqin granules group was given gastric gavage of Chanqin granules (9.36 g·kg-1·d-1)after PM2.5 exposure,Huifeining group was given gastric gavage of Huifeining (8 mL·kg-1·d-1)after PM2.5 exposure,and saline control group was given gastric gavage of normal saline after intratracheal drop infusion of normal saline , the treatment covering 2 weeks. After medication,the contents of substance P (SP),calcitonin gene related peptide(CGRP), neurokinin A(NKA), neurokinin B(NKB), and nerve growth factor(NGF) in rat serum were detected by enzyme-linked immunosorbent assay (ELISA), the expression levels of SP and NGF in lung tissue and dorsal root ganglia were measured by immunohistochemistry , and the protein and mRNA expression levels of NGF in lung tissue and dorsal root ganglia were determined by Western blotting method and reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR),respectively. Results Compared with the saline control group, the levels of SP, NKA, NKB, NGF and CGRP in rat serum, immunohistochemical mean optical density of NGF in lung tissue and SP and NGF in dorsal root ganglia, the protein and mRNA expression levels of NGF in lung tissue and the protein expression level of NGF in dorsal root ganglia of PM2.5 exposure group were all increased(P < 0.05). Compared with PM2.5 exposure group,the above indexes in anti-NGF group, Chanqin granules group, and Huifeining group were decreased (P < 0.05), and there being no differences between the 3 groups (P > 0.05). Conclusion Chanqin granules have certain effects on relieving the neurogenic inflammation in rat airway induced by PM2.5 through inhibiting the expression of NGF.

Objective To study the changes of amplitude of low-frequency fluctuation (ALFF) in blood oxygenation level dependent functional MRI (BOLD-fMPI) in non-lesional epilepsy (NLE),and discuss its underlying neurophysiological mechanism. Methods The BOLD-fMRI data of 16 patients with NLE and 15 normal volunteers were analyzed by ALFF. The amplitude of the blood oxygenation level dependent activation of the resting state brain was investigated. The brain structures showing increased and decreased ALFF in NLE patients were demonstrated by comparing to normal subjects with 2-sample t-test with threshold of P＜0.05. Results As compared with those in normal subjects,the regions showing increased ALFF in NLE patients were distributed in the right temporal lobe (Montreal Neurological Institute [MNI] coordinates:x=15,y=90,z=21),medial frontal lobe (MNI coordinates:0,24,-24),ventral anterior cingulated (MNI coordinates:-12,30,27) and right cerebellar hemisphere (MNI coordinates:-51,-57,-4); while the regions showing decreased ALFF covered the areas of the left cerebellar hemisphere (MNI coordinates:-48,-15,39),posterior cingulum gyrus (MNI coordinates:60,-21,33) and precuneus (MNI coordinates:-6,-54,66). Conclusion NLE patients show abnormal brain functional organization in resting state; the increased ALFF is considered to be the facilitation such as epileptic activity generation and propagation,while the decreased ALFF might be considered as the functional inhibition in these regions.

OBJECTIVETo investigate the correlation between impaired non-viral specific immune function of dendritic cell (DC) and viral clearance and cytotoxic T lymphocyte (CTL) response to HBV or HCV in patients with HBV and HCV coinfection.

METHODSTwenty-five patients with HBV and HCV coinfection were investigated in this study. In 1994 and 2002, biochemical and virological markers and quantitative serum HBV DNA and HCV RNA levels were detected in these patients. According to the virus clearance status, these patients were divided into 4 groups: 14 patients with both HBV and HCV clearance (Group A), 6 patients with HCV clearance only (Group B), 3 patients with HBV clearance only (Group C), and 2 patients with persistent infection of HBV and HCV (Group D). Phenotypes and immune functions of monocyte-derived DCs were compared between these groups. 51Cr release assay were used to measure CTL response to epitopes derived from HBV, HCV or influenza virus (as positive control) in HLA-A2+ patients.

RESULTSImpaired non-viral specific immune functions of DCs were observed in group B, C and D compared with group A and normal donors (Group N). These impaired functions included CD86 decreasing expression and lower capacity to stimulating allogenic T cells and uptaking antigen. The specific CTL response to HBV- and HCV-derived peptides could be induced in group A (12/12). The specific CTL response to HBV-derived peptides or to HCV-derived peptides could be induced in group C (3/3) or B (5/5), respectively. But the specific CTL response to both of two HBV-derived peptides or two HCV-derived peptides could not be induced in group C (0/3) or B (0/5), respectively. And no CTL response to HBV or HCV-derived peptides could be induced in groups D (0/1) and N (0/4).

CONCLUSION1. The results suggest that specific CTL response to HBV or HCV play a vital role in the viral clearance. 2. The DCs with impaired non-viral specific immune functions exist in chronic patients with HBV and/or HCV infection, but do not interfere with clearance and CTL response to HBV or HCV. It is reasonable to speculate that impaired functions of DCs result from viral infection.

Adult ; Dendritic Cells ; immunology ; Female ; Hepacivirus ; immunology ; Hepatitis B virus ; immunology ; Humans ; Immunophenotyping ; Lymphocyte Culture Test, Mixed ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo evaluate the method and effectiveness of rigid-bronchoscopic cryosurgery for advanced central lung cancer.

METHODSForty-eight patients were enrolled in this study from June 2002 to December 2008, including 33 male and 15 female. The average age was 70 years (ranged from 45 to 83 years old). For the 48 patients, 38 cases were patients with advanced central lung cancer who were not suitable for surgery, and the remaining 10 cases were patients with local recurrence in trachea or main bronchus postoperatively. Cryosurgery was performed 120 times for all patients, 2.5 times per patient on average. The trachea or bronchus station, symptom such as dyspnea, hemoptysis, respiratory function and quality of life were observed.

RESULTSThe unblocked ratio of trachea and bronchi was 97%. All patients got satisfied improvement ratio of symptoms, 87.5% for dyspnea, 72.9% for cough, 93.8% for hemoptysis and 62.5% for chest pain. Respiratory function tests showed that both the mean forced expiratory volume in first second and forced vital capacity got an improvement from (1.03+/-0.05) L to (1.85+/-0.13) L and from (1.69+/-0.18) L to (2.96+/-0.14) L respectively (P<0.01). Karnofsky score also got no less than 20 scores improvement. The Follow-up time was 6 to 62 months. The longest survival was 62 months. The median survival time was 20 months. There was no severe perioperative complications and mortality except for 3 cases of moderate exeduation.

CONCLUSIONSCryosurgery is easy to perform with minimal complications. Not only could it provide an effective and rapid control of symptoms caused by central lung cancer, it could also unobstructed bronchus promptly and improve patients' quality of life.

Aged ; Aged, 80 and over ; Bronchoscopy ; Cryosurgery ; methods ; Female ; Humans ; Lung Neoplasms ; surgery ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo study whether dendritic cells (DCs) derived from the peripheral blood in chronic hepatitis B patients can induce specific T cell immune response.

METHODS(1)The subjects were divided into 3 groups: chronic hepatitis B group (CHB), acute hepatitis B group (AHB), and normal donor group (ND). The peripheral blood mononuclear cells (PBMCs) isolated from those subjects were stimulated with HBcAg 18 to 27 CTL epitope peptide, and intracellular cytokine staining (ICCS) was used for detecting IFN-gamma, IL-2 and TNF-alpha produced by CD8+ T cell. (2) DCs generated from PBMCs were pulsed with HBcAg 18 to 27 CTL epitope peptide, then were cocultured with autologous lymphocytes for 10 days to induce antigen-specific T cell, which was assessed by ICCS and cytotoxic assay.

RESULTS(1) The memory effect of the PBMCs from AHB group to HBcAg 18 to 27 CTL epitope peptide was stronger than that from CHB or ND group (t=2.508-3.305, P<0.05). (2)After lymphocytes were cocultured with DC treated with HBcAg 18 to 27 CTL epitope peptide, antigen-specific T cell effect was induced. And the killing rates were (57.0+/-23.0)%, (49.5+/-20.2)%, (21.8+/-12.9)% at the effector/target of 30:1, 10:1, 3:1, which were higher than that in control group.

CONCLUSIONSThe memory T cells against HBV antigen lacks in CHB patients. DCs from CHB patients pulsed with HBcAg 18 to 27 epitope peptide can induce HBV antigen-specific T cell, which can kill specific target cells and produce cytokines involved in virus clearance.

Adult ; CD8-Positive T-Lymphocytes ; immunology ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; virology ; Epitopes, T-Lymphocyte ; immunology ; Female ; Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China.

METHODSUsing polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China.

RESULTSComparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang.

CONCLUSIONThe results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar.

China ; DNA, Viral ; genetics ; Genotype ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; Serotyping

Objective To evaluate the effect of surgical excision of the anterior temporal lobe and hippocampus on the cognitive function of patients with refractory temporal lobe epilepsy. Methods Nineteen patients with refractory temporal epilepsy received surgical excision of the anterior temporal lobe and hippocampus, and cognitive neuropsychologieal assessments were conducted, 3 and 6 months after the surgery, to evaluate the changes in the patinets' cognitive functions. Results Satisfactory effects were achieved in these patients after the operations. One patient showed temporary speech disorder, and two exhibited temporary euphoria. Patients with refractory temporal lobe epilepsy had severe cognitive dysfunctions involving especially in the intelligence, attention and memory, which were not aggravated after the operation, Conversely, the operation resulted in gradual improvement of some of the cognitive functions in these patients. Conclusion Patients with refractory temporal lobe epilepsy have cognitive dysfunction, and surgical excision of the anterior temporal lobe and hippocampus can cure or reduce seizure of temporal lobe epilepsy and improve the cognitive dysfunctions to some extent.

Objective To study the impairnlent of cognitive function and the change of P300 event-related potential(ERP)parameters in the patients with tumor in frontal lobe so as to investigate the role of the front lobe in cognitive function and the P300 production. Methods P300 examination and neuropsychological tests for cognition,including Stroop Test,Symbol Digit Modalities Test,and Verbal Fluency Test,et al,were performed in 31 patients with tumor in front lobe(left 15 and right 16)and 30 healthy controls. Results Compared with healthy controls, in the patients with front lobe tumor,cognitive function was degraded(P＜0.05),the latency of N3,P3 of P300 was significantly longer,and the amplitude of P3 was significantly lower(P＜0.05).Funhermore,the patients with tumor in right front lobe had signmcantly lower scores in cognitive tests compared with healthy controls (P＜0.05), but the ones with tumor in left front lobe were demonstrated obviously impaired only in Verbal Fluency Test(P＜0.05).Compared with healthy controls, the patients with tumor in eimer right or left flront lobe, the latency of N3,P3 of P300 was significantly longer,the amplitude of P3 was significantly lower(P＜0.05),while in the comparison of two sides, there was no statistically significantly difference in P300 (P＞0.05).Conclusions The cognitive function is obviously impaired in the patients with tumor in frontal lobe,and the impainnent is more serious in the ones with right frontal lobe tumor. Frontal lobe may be related to the generation and transmission of P300,and it is not significantly different in between left and right frontal lobes.