OBJECTIVETo explore the effect and mechanism of cinnabaris and realgar in promoting awake effect of endotoxin- induced brain injury rat applied with Angong Niuhuang Wan.

METHODNormal rats implanted cortical electrode in advance were divided into 6 groups: control, model, the Angong Niuhuang Wan (AGNH, 0.4, 0.2 g · kg(-1)), the Angong Niuhuang Wan without cinnabaris and realgar (QZX-AGNH, 0.32, 0.16 g · kg(-1)). Rats in the control and model groups were given distilled water. After three days of intragastric administration, the brain injury model was injected with endotoxin through tail vein. Then trace electro-corticogram (EcoG) 1-6 h after LPS injection, and compare the power and relative power of beta (β) and delta-waves (δ) at 6 h of these groups. The content of acetylcholine (Ach) and the affinity of M-receptor (M-R) in cortex and brainstem were detected by alkaline hydroxylamine colorimetric method and radioactive ligand binding assay, respectively.

RESULTAGNH (0.4, 0.2 g · kg(-1)) could increase the power and relative power of β and AGNH (0.4 g · kg(-1)) showed better action on brain electrical activation. QZX-AGNH showed weak effect on it. AGNH (0.4 g · kg(-1)) could increase the affinity of M-R in cortex and the content of Ach in brainstem. The action of QZX-AGNH was not obvious.

CONCLUSIONIn endotoxin-induced brain injury rats, AGNH can raise the cholinergic system function of cortex, and strengthen the uplink of cortex activation of brainstem cholinergic system, improve the level of cortical activity and enhance the activation of EcoG to promote the body's awakening. QZX-AGNH show weak effect. Cinnabaris and realgar play an important role in promoting awake effect in endotoxin-induced brain injury applied with Angong Niuhuang Wan. The mechanism may be related to cortical and brainstem cholinergic system function.

Animals ; Brain Injuries ; chemically induced ; drug therapy ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; Endotoxins ; adverse effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo reveal the transmembrane signal pathway participating in regulating neuron functions of treating Alzheimer's disease (AD) by acupuncture.

METHODSSAMP8 mice was used for AD animal model. The effect of acupuncture method for qi benefiting, blood regulating, health supporting, and root strengthening on the amount and varieties of transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice was detected using HPLC MS/MS proteomics method.

RESULTSCompared with the control group, acupuncture increased 39 transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice, of them, 14 belonged to ionophorous protein, 8 to G protein, 8 to transmembrane signal receptor, and 9 to kinase protein. Totally 3 main cell signal pathways were involved, including G-protein-coupled receptors signal, enzyme linked receptor signal, and ion-channel mediated signal. Compared with the sham-acupuncture group, acupuncture resulted in significant increase of kinase signal protein amount. From the aspect of functions, they were dominant in regulating synapse functions relevant to cytoskeleton and secreting neurotransmitters.

CONCLUSIONThe cell biological mechanism for treating AD by acupuncture might be achieved by improving synapse functions and promoting the secretion of neurotransmitters through transmembrane signal transduction, thus improving cognitive function of AD patients.

Acupuncture Therapy ; Alzheimer Disease ; metabolism ; Animals ; Disease Models, Animal ; Female ; Male ; Membrane Microdomains ; metabolism ; Mice ; Proteomics ; Signal Transduction ; Tandem Mass Spectrometry

Objective To investigate the analgesic efficacy of intrathecal injection of RC-13,a competitive kinesin superfamily protein 17 antagonist,in a mouse model of bone cancer pain.Methods Forty male C3H/HeJ mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 5 groups (n=8 each): sham operation group (group S); bone cancer pain + 5 μl dimethyl sulfoxide (DMSO) group (group R0); bone cancer pain + 2.5 μg RC-13 group (group R1); bone cancer pain + 5 μg RC-13 group (group R2) and bone cancer pain + 10 μg RC-13 group (group R3).In groups R0-3,bone cancer pain was induced by implantation of α-min-imal essence medium (α-MEM) containing osteosarcomaNCTC 2472 cells into the intramedullary space of right femur.In group S,culture medium α-MEM containing no cancer cell was injected instead.10％ DMSO 5 μl and RC-13 2.5 μg/5 μl,5μg/5μ1 and 10 μg/5 μ1 dissolved in 10％ DMSO were injected intrathecally in groups R0-3,respectively,once a day for 3 consecutive days starting from 14th day after inoculation of the tumor cells.Pain behavior was assessed by the paw withdrawal mechanical threshold (PWMT) and spontaneous lifting times (SLTs) measured at 1 day before inoculation and at 3,5,7,10,14 days after inoculation.The same tests were also performed at 1,3,5 and 7 days after administration in groups R0-3.Results Compared with group S,PWMT was significantly decreased and SLTs were increased at 7-14 days after inoculation in the other groups (P ＜ 0.05).Compared with group R0,PWMT was significantly increased and SLTs were reduced at 1 day after administration in group R1,at 1and 3 days after administration in group R2,and at 1,3 and 5 days after administration in group R3 (P ＜ 0.05).Compared with group R1,PWMT was significantly increased and SLTs were reduced at 3 days after administration in group R2,and at 1,3 and 5 days after administration in group R3 (P ＜ 0.05).Compared with group R2,PWMT was significantly increased and SLTs were reduced at 1 and 3 days after administration in group Rs (P ＜ 0.05).Conclusion Intrathecal RC-13,a competitive kinesin superfamily protein 17 antagonist,has a good analgesic efficacy in a mouse model of bone cancer pain and the efficacy is dose-dependent.

A diving decompression procedure is a specific rule that divers should follow when they ascend and get out of water. It comes from the decompression theory and algorithm and is designed for the prevention of decompression sickness. With the ， ， development of diving technology and diving medicine the decompression procedures are constantly innovated and the new ， decompression procedure can be used in diving practice after safety verification. In principle the safety verification of ， decompression procedures should be conducted on animal experiments before human experiments and the risks of ， decompression sickness and oxygen toxicity should be systematically assessed. However the assessment methods used in ， ， ， different studies differ greatly thus it is urgent to establish a standard and universal verification system. Traditionally the risk ， ， assessment of decompression sickness and oxygen toxicity is mainly carried out by observing the incidence detecting bubbles ， theoretical calculation and lung functional test. Furthermore biochemical indicators are increasingly becoming important ， ， supplements. Due to the special underwater environment the diving operation is prone to accidents. Therefore in addition to ， verifying the safety of the new decompression procedure exploring its safety decompression limit is of great significance for the formulation of emergency decompression procedures in emergency situations. The specific approach is to shorten the decompression time and assess the safety until the critical time for detecting bubbles without the occurrence of decompression ， ， sickness is found. Future studies should continue to optimize safety assessment methods explore sensitive biochemical markers ， clarify species associations and improve verification efficiency and reliability of results.

BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26％ of cells were positive for CD29,7. 51％ for CD34 and 4. 02％ for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.

Objective To analyze the value of plane QRS-T angle on prediction of malignant ventricular arrhythmia (MVA) occurred after emergency percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods The clinical data of 418 patients with STEMI who underwent PCI within 12 h of symptom onset were retrospectively analyzed, and the patients were divided into plane QRS-T angle ≤ 90° group (324 cases) and plane QRS-T angle>90° group (94 cases) according to the plane QRS-T angle after PCI. The clinical data were compared between 2 groups. Results Compared with patients in plane QRS-T angle ≤ 90° group, patients in plane QRS-T angle > 90° group was older: (67.4 ± 11.8) years vs. (63.6 ± 12.0) years, QTc interval was longer: (438.60 ± 34.97) ms vs. (425.24 ± 25.49) ms, rate of left ventricular ejection fraction (LVEF) <45% was higher: 57.4% (54/94) vs. 35.8% (116/324), rate of using of beta-blockers was less: 74.5% (70/94) vs. 84.9% (275/324), but the incidences of hypertension and MVA were higher:79.8%(75/94) vs. 64.5%(209/324) and 10.6%(10/94) vs. 1.2%(4/324), and there were statistical differences (P<0.01 or<0.05). Logistic regression analysis showed that plane QRS-T angle >90° was an independent risk factor of MVA after PCI in STEMI patients (OR = 9.640, P =0.001), and using of beta-blockers was a protective factor (OR = 0.266, P = 0.028). Conclusions Plane QRS-T angle>90° is an independent risk factor of MVA after PCI in STEMI patients, while the use of beta-blockers is a protective factor. Paients with STEMI after PCI should be alert to the occurrence of MVA in the condition of plane QRS-T angle>90° and not taking beta-blockers.

Objective To understand the integrated ability of parasitic disease prevention and control of professional person-nel of Jiangsu Province through the contest. Methods Totally 56 players from the whole province were selected,and all the players participated in the contest. The theory knowledge and skill scores were collected and the statistical analyses were con-ducted. Results The average theoretical score of the participants was 88.86±15.56 and the passing rate was 91.1％. The aver-age skill operating score was 69.16 ± 16.01 and the passing rate was 67.9％. The average Plasmodium microscopy score was 16.54±8.09 and the passing rate was 50％. The average helminth egg microscopy score was 34.27±10.66 and the passing rate was 67.9％. There were statistical differences among the age groups and different levels of schistosomiasis endemic situation (F =5.10,6.39,both P<0.01). The theoretical knowledge including schistosomiasis,malaria,hydatid disease and others and the score rates were 91.07％,90.94％,85.83％and 90.93％,respectively. The hydatid disease score rate was lower(χ2=19.17, P<0.01). The radar chart displayed that the score rates of tabletting and microscopy test in Kato-Katz film production ,malaria blood film production and microscopy test were all low. Conclusion In Jiangsu Province,the participants have higher score in the theory test. However,they have lower skill test score,especially in the parasite species identification. The operational skills still need to be strengthened for center for disease control(CDC)participants.

DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.
Base Sequence ; Chloroplasts ; genetics ; DNA Barcoding, Taxonomic ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Liliaceae ; genetics ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo probe the mechanism of EEG activation and Xingnao Kaiqiao, evaluate the actions of cinnabaris and realgar in Xingnao Kaiqiao of Angong Niuhuang pill, guess the significance of cinnabaris and realgar in specific indication treatment of Angong Niuhuang pill, and provide experimental bases for the rationality of Angong Niuhuang pill building-up.

METHODSeventy SD rats were divided into seven groups: the control, the model, the Angong Niuhuang pill (0.4 g x kg(-1)), the Angong Niuhuang pill without cinnabaris and realgar (0.32 g x kg(-1)) , the cinnabaris and realgar (0.08 g x kg(-1)), the realgar (0.04 g x kg(-1)), and the cinnabaris (0.04 g x kg(-1)). Rats in the control and model groups were given distilled water. After three days of administration, the brain damage model was made by Lipopolysaccharides (LPS) injection through caudal vein and the catecholamine (CA) and its metabolites levels in cerebral cortex, included noradrenaline (NE), adrenaline (E), 3-methocy-4-hydroxyphenylglycol (MHPG), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), dopamine (DA), Homovanlic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC), were determined by high-performance liquid chromatography with electrochemical detector (HPLC-ECD). Influences of Angong Niuhuang pill, Angong Niuhuang pill without cinnabaris and realgar, cinnabaris and realgar on monoamine transmitters were observed in brain damage rats caused by LPS.

RESULTLPS could raise NE, 5-HT, 5-HIAA levels and reduce E, DOPAC levels, but had no influence on HVA, DA, MHPG levels. Angong Niuhuang pill had the trend of raising E, DOPAC levels and reducing NE level, and could reduce 5-HIAA level obviously comparing with models. But Angong Niuhuang pill without cinnabaris and realgar was different, NE level was significantly higher compared to models and Angong Niuhuang pill, DA level was also significantly higher compared to all groups. Cinnabaris and realgar had the same action trends with Angong Niuhuang pill, and separate realgar could obviously reduce 5-HT.

CONCLUSIONInfluence on CA and its metabolites levels in cerebral cortex may be one of the mechanisms of Angong Niuhuang pill's EEG activation, and cinnabaris and realgar have the same action on CA levels in cerebral cortex. The results of the present work allow us to put forward the hypothesis that cinnabaris and realgar are most likely one of the important material basis in Xingnao Kaiqiao of Angong Niuhuang pill.

Animals ; Arsenicals ; pharmacology ; Brain Injuries ; chemically induced ; metabolism ; physiopathology ; Catecholamines ; metabolism ; Cerebral Cortex ; metabolism ; physiopathology ; Drug Combinations ; Electroencephalography ; drug effects ; Lipopolysaccharides ; Male ; Medicine, Chinese Traditional ; Mercury Compounds ; pharmacology ; Norepinephrine ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Sulfides ; pharmacology

BACKGROUNDBioreactors are pivotal tools for generating mechanical stimulation in functional tissue engineering study. This study aimed to create a bioreactor that can simulate urinary bladder mechanical properties, and to investigate the effects of a mechanically stimulated culture on urothelial cells and bladder smooth muscle cells.

METHODSWe designed a bioreactor to simulate the mechanical properties of bladder. A pressure-record system was used to evaluate the mechanical properties of the bioreactor by measuring the pressure in culture chambers. To test the biocompatibility of the bioreactor, viabilities of urothelial cells and smooth muscle cells cultured in the bioreactor under static and mechanically changed conditions were measured after 7-day culture. To evaluate the effect of mechanical stimulations on the vital cells, urethral cells and smooth muscle cells were cultured in the simulated mechanical conditions. After that, the viability and the distribution pattern of the cells were observed and compared with cells cultured in non-mechanical stimulated condition.

RESULTSThe bioreactor system successfully generated waveforms similar to the intended programmed model while maintaining a cell-seeded elastic membrane between the chambers. There were no differences between viabilities of urothelial cells ((91.90 ± 1.22)% vs. (93.14 ± 1.78)%, P > 0.05) and bladder smooth muscle cells ((93.41 ± 1.49)% vs. (92.61 ± 1.34)%, P > 0.05). The viability of cells and tissue structure observation after cultured in simulated condition showed that mechanical stimulation was the only factor affected cells in the bioreactor and improved the arrangement of cells on silastic membrane.

CONCLUSIONSThis bioreactor can effectively simulate the physiological and mechanical properties of the bladder. Mechanical stimulation is the only factor that affected the viability of cells cultured in the bioreactor. The bioreactor can change the growth behavior of urothelial cells and bladder smooth muscle cells, resulting in the cells undergoing adaptive changes in mechanically-stimulated environment.

Bioreactors ; Cell Line ; Humans ; Myocytes, Smooth Muscle ; cytology ; Tissue Engineering ; methods ; Urinary Bladder ; cytology ; Urothelium ; cytology