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Gold nanochannels were prepared using Al2 O3 nanotubules membrane as the carrier and modified with chitosan by a classical N-(3-dimethylaminopropyl)-N-ethyl carbodiimide ( EDC)/N-hydroxysuccinimide ( NHS ) coupling reaction. The nanochannels were characterized by field emission scanning electron microscopy ( FESEM) , cyclic voltammetry and AC impedance method. The Au nanochannels modified with chitosan showed a chiral environment and can be used to separate histidine enantiomer. The effects of pore size and solution pH on the separation efficiency of histidine were investigated. To increase the detection sensitivity of D-, L-histidine, Ag nanoparticles were used to enhance the surface enhanced Raman scattering ( SERS) activity. The results showed that the chitosan-modified gold nanochannels can be used to separate chiral histidine based on this unique selective nanochannel membrane. L-Histidine and D-histidine were respectively detected by SERS at wavelengths of 1000 and 1590 cm-1 . The results showed that L-histidine and D-histidine were separated well in the mixture containing 200 μL of histidine, 100 μL of colloidal Ag and 100 μL of 80 mmol/L NaCl ( pH=7 . 59 ) with a separation efficiency of 4 . 91 .
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AIM: To observe the kinetic alteration of nitric oxide formation in the lungs in the development of pulmonary fibrosis in the rat. METHODS: The contents of hydroxyproline in the lungs, NO-2/NO-3 (nitrite/nitrate) in out-flowing and in-flowing pulmonary blood (OPB, IPB) were assayed on the day 7, 14, 21, 30 and 70 after intratracheal administration of bleomycin A5 . The content of NO-2/NO-3 in supernatants of culture of the alveolar macrophages (AMs) and the amount of iNOS positive stain cells in lung tissue section were also observed in the rat on 14th day after-bleomycin A5 administration. RESULTS: The content of lung hydroxyproline had no change on the 7th day, increased on the 14th day (P＜0.05), increased significantly on the 21th day, 30th day and 70th day post-bleomycin A5 compared with control rats. On the 7th day and 14th day, the content of NO-2/NO-3 increased in OPB and decreased in IPB (P＜0.01). On the 21th day, the content of NO-2/NO-3 abated in OPB (P＞0.05) but still decreased in IPB (P＜0.01). On the 30th day and the 70th day, the NO-2/NO-3 level recovered both in OPB and IPB. AMs from rats on the 14th day post-bleomycin A5 showed significant elevation (P＜0.01) in NO-2/NO-3 level. The amount of iNOS positive stain cells increased in rats on the 14th day post-bleomycin A5. CONCLUSION: The amount of NO in the lungs was high in the initial phase of fibroproliferative reaction induced by bleomycin A5 ,and these might be associated with the enhanced ability of AMs to release NO and the increased amount of iNOS.
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AIM: To study the up-regulation of inducible nitric oxide synthase (iNOS) in lung of pulmonary fibrosis and its relationship with fibrosis. METHODS: The changes of amount of iNOS positive stain cells and type Ⅰ?Ⅲ collagen were examined on the day 7, 14 and 30 after intratracheal administration of bleomycin A_5. The contents of NO-_2/NO-_3 (nitrite/nitrate) in out-flowing pulmonary blood (OPB), hydroxyproline in lung and the histological changes were detected after iNOS was blocked by aminoguanidine (AG). RESULTS: (1) The number of iNOS-positive stain cells increased significantly in BLMA_5 7 d, 14 d and 30 d groups compared with that in control group (P
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AIM: To investigate the effects of nitric oxide on ultrastructure and anionic sites of glomerular in renal ischemia reperfusion injured(I-RI) rats.METHODS: Animals were divided randomly into five groups:(1) sham group(n=6);(2) I-RI group(n=6),0.3 mL normal saline was injected via venae lingualis 20 min before ischemia;(3) SNP+I-RI group(n=6),2.5 ?g/kg sodium nitroprusside(SNP) was injected via venae lingualis 20 min before ischemia;(4) AG+I-RI group(n=6),10 mg/kg aminoguanidine(AG) was injected via venae lingualis 20 min before ischemia;(5) L-NNA+I-RI group(n=6),10 mg/kg N~?-nitro-L-arginine(L-NNA) was injected via venae lingualis 20 min before ischemia.Anionic sites of glomerular were studied with a cationic probe-polyethyleneimine(PEI) and ultrastructure was observed under electron microscope in renal I-RI rats.RESULTS:(1) Ultrastructure of glomerular was normal and anionic sites(AS) was located clearly in lamina rare externa of GBM in sham rats.The PEI particles arranged regularly in line(19.3?1.7/(1 000 nm)) under electronic microscope.Obvious foot processes derangement and effacement were observed and the AS number in GBM of I-RI group was fewer(16.6?1.0/(1 000 nm),P0.05).CONCLUSION: Foot process effacement and reduction of anionic sites were present in glomerular filtration membrane in renal I-RI rats.NO aggravated those injuries,indicating that NO plays a role in the ultrastructure damages of glomerular filtration membrane in I-RI rats.
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AIM: To observe the kinetic alteration of nitric oxide formation in the lungs in the development of pulmonary fibrosis in the rat. METHODS: The contents of hydroxyproline in the lungs, NO - 2/NO - 3 (nitrite/nitrate) in out-flowing and in-flowing pulmonary blood (OPB, IPB) were assayed on the day 7, 14, 21, 30 and 70 after intratracheal administration of bleomycin A 5 . The content of NO - 2/NO - 3 in supernatants of culture of the alveolar macrophages (AMs) and the amount of iNOS positive stain cells in lung tissue section were also observed in the rat on 14th day after-bleomycin A 5 administration. RESULTS: The content of lung hydroxyproline had no change on the 7th day, increased on the 14th day ( P 0.05) but still decreased in IPB ( P
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AIM: To observe the changes in proliferation and apoptosis of macrophages in the development of pulmonary fibrosis in rats. METHODS: The number of macrophages, apoptotic cells, the proliferation index (PI) and MTT activity of macrophages were assayed on the day 14 and the day 30 after intratracheal adminstration BLMA_5 in rats.RESULTS: (1) The number of alveolar macrophages was increased in BLMA_5 14 d group and in BLMA_5 30 d group, compared with sham 14 d group and sham 30 d group, respectively. The number of macrophages in BLMA_5 14 d group was higher than that in BLMA_5 30 d group. (2) The PI of macrophages increased in BLMA_5 14 d group, and decreased in BLMA_5 30 d group. (3) The number of apoptotic cells increased both in BLMA_5 14 d group and BLMA_5 30 d group.The number of apoptotic cells in BLMA_5 14 d group was lower than that in BLMA_5 30 d group. (4) The MTT activity of macrophages was higher in BLMA_5 14 d group and in BLMA_5 30 d group than that in sham 14 d group and sham 30 d group, respectively. CONCLUSIONS: The ability of proliferation increased at first, and then decreased, but the apoptosis of macrophages increased all the time, in the development of pulmonary fibrosis. This might be partly contributed to the changes of the number and function of macrophages in lung.
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AIM: To investigate the urinary protein (UP) excretion in rats suffering from renal ischemia-reperfusion (I-R) and effect of nitric oxide on it. METHODS: SD rats were used to establish the renal I-R model. Sodium nitroprusside (SNP), N ?-nitro-L-arginine (L-NNA) and aminoguanidine (AG) were used to determine the effect of nitric oxide on UP excretion under renal I-R. Quantitative analysis of UP was made by chromatometry. UP species were separated by SDS-PAGE. RESULTS: Renal I-R caused significant increase in UP ( P
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AIM: To study the role of polymorphonuclear neutrophile(PMN) in lipopolysaccharide (LPS)- induced acute lung injury (ALI) and the protective effect of interleukin - 10(IL - 10) on ALI. METHODS: LPS alone (100 ?g) or LPS+ IL-10 (l ug) was instilled intratracheally into rats. PMN numbers, protein content and malondialdehyde (MDA) content in bronchoalveolar lavage fluid (BALF) were measured. Histological change of lung was also observed. RESULTS: LPS increased significantly PMN numbers, protein content and MDA content in BALF. Histological finding shows PMN accumulation in lung. IL - 10+LPS reduced remarkably PMN numbers ,pro- tein content and MDA content in BALF than those caused by LPS. PMN decreasing was also identified by light microscopy. CONCLUSION: LPS instilled intratracheally causes PMN accumulation in lung and ALI, while IL - 10 could alleviate ALI through reducing PMN accumulation.
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Endotoxin of in-and out-flowing pulmonary blood (IPB ,OPB) was measured by limulus chromogenic test in rabbits; Clearance of ~(51)Cr-E. coli endotoxin by lung was measured by a double indicator dilution method in rabbits; 12 and 24 hour's mortality rates was observed after E. coli endotoxin (1?g/100g b. w.) was infused through vein (ⅳ)and aorta (av)into the lead-sensitized rats. The endotoxin level of IPB (44.45?31.73 pg/ml plasma, n=13) was significantly higher than that of OPB (19.23?17.85 pg/ml plasma, n=13)in normal rabbits, (P
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The concentrations of CuZn-SOD and malondialdehyde (MDA) in in-flowingand out-going pulmonary blood(IPB, OPB) were observed dynamically during intestinal ischemic-reperfusion in rabbits. The results showed that in normal subjects the content of SOD of OPB was higher than IPB, P
ABSTRACT
Effective analgesia was induced by electro-acupuncture stimulating the "Zusanli"(足三里) point in the rats. The animal were sacrificed and as soon as possible and the median eminence were taken out from the brian. The specimens were fixed in glutaraldehyde, then were prepared for electron microscope observation.Twelve male adult rats were divided into control and experimental groups. There were many terminal enlargements in the fibrous zone of rat's median eminence. Different types of vesicles were found in the terminal buttons, they were round clear and flattened clear vesicles, and large or small vesicles with a dense core. There were also mixed forms, and irregular vesicles in the same button. Synapses may be axo-dendrits, axo-axon or axe-somatic synaptic types.The neurous in the fibrious zone could be classified into "light" and "dark" cells. The light cells might be the neurosecretory neurons. The neuroglia cells were oligodendrocytes and astrocytes.In acupuncture analgesia experimental group the terminal enlargment contained less number of round clear vesicles than that of the control group, and sometime they were empty. This may indicate the discharge of acetylcholine, to increase the secretion of the neurosecretory substance and enkephalin. But the flattened clear vesicles had no change in the terminal button. The neurons and neroglia also exhibited some morphological changes which may indicate the cells were in active functional state.
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Twenty adult male rats, weighing 200～300 g were divided into 3 groups: (1) Six. animals were stimulated through two electrodes fixed on the tail to produce somatic pain.(2) Six animals were stimulated by increasing pressure in the stomach through an air balloon to cause visceral pain. And 8 animals served as control.All the animals were sacrificed and their neurohypophysis were fixed in 4％ glutaraldehyde and 1％ OsO_4 successively. Their section were cut with an LKB microtome and observed under H 500 electron microscope.The experimental results summarized as follows:1. The bodies of pituicytes under the pain stimulation were hypertrophied. The rough endoplasmic reticulum, polyribosomes and mitochondria were increased. The Golgi complex was well developed. There were numerous elliptical vesicles appearing in the cytoplasm of pituicytes.2. The number of large lipid masses was increased while the processes of pituicytes engulfed the degenerative neurosecretory terminal into them and the pituicyte digted it finally.3. Neurosecretory granules were decreased but the synaptic small vesicles were increased in the dilated parts of neurosecretory fibers after the pain stimulation.4. The continuous network of perivascular space and interstitial space between the dilated parts of neurosecretory fibers were widened.
ABSTRACT
The purpose of this research work is to study the influence of experimentally produced pain on the uhrastructural changes of the nerve endings of the neurosecretory fibers from hypothalamus to neurohypophysis.Twenty adult male rats, weighing 200～300 g were divided into 3 groups: (1) Six animals were stimulated through two electrodes fixed on the tail to produce somatic pain. (2) Six animals were stimulated by increasing pressure in the stomach through an air balloon to cause visceral pain. And 8 animals served as control.All the animals were sacrificed and their neurohypophysis were fixed in 40％ glutaraldehyde and 1％ OsO_4 successively. Sections were cut with an LKB microtome and observed under H500 electron microscope.The following conclusions were drawn from the observed results:1. The neurosecretory granules in the fibers from hypothalamus to neurohypophysis grew up to maturity by increasing in size gradually as they passing down from hypothalamus to neurohypophysis.2. Basing on the morphological exhibition of the neurosecretory granules, we get the impression that the neurosecretion might be released either by molecular dispersion or by exocytosis.3. There were a great number of discrete smooth vesicles grouped in cluster in the dilated parts of neurosecretory fibers. These vesicles had been called as "synaptic" or "synaptoid" vesicles. They might be really synaptic vesicles and containing acetylcholine. Other vesicles with irregular profile and varying in size, collapsing vesicles and vacuoles might be the residues of the neurosecretory granules after discharging their content.4. Many synaptic vesicles were concentrated at the axolemma of the dilated parts of neurosecretory fibers and the membranes were thickened and leaving a synaptic cleft.5. In the exprimental groups after pain stimulation the neurosecretory granules decreased in number and the synaptoid vesicles increased in number and the neurosecretory granules showed a wide pale halo. The number of irregular vesicles and ghost of granules were increased. This phenomenon might show that the excretion of neurosecretory material and neurotransmitters was enhanced.6. Neurohemal zones were the barrier for the neurosecretory material entering into blood. Acetylcholine, endogenous opiatelike substances and gamma-aminobutyric acid (GABA) might play a control role for neurosecretion at this site.7. This experimental results supported the action mechanism of endogenous opaitelike substance and morphine could produce an important role directly in neurohypophysis.
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Our experiments were designed to observe the cytological changes of the anterior lobes in rats pituitary at electron microscopic level after electric pain-stimulation. obvious changes were founded in the ACTH, GH and PRL cells and slight changes in the FSH and TSH cells. However, the LH cells remained to resemble the normal cells. In the experiment group of the somatic painful stimulation the results were as follows: The ACTH cells were irregularly shaped, with extended thin and long plasmatic processes which interdigitated with GH cells. The processes filled with secretory granules. Some of the Secretory granules aligning along the plasmatic membrane showed pale density. The flat cisternae of rough endoplasmic reticulum (RER) appeared to be slightly dilated. The sizes of the bodies of GH and PRL cells increased. In the GH cells there were numera media cisternae in RER, their Golgi complex was well developed, and the secretory granules were distributed to the periphery of the cell bodies. All these phenomenous showed the secretory activity of these cells was enhanced. The PRL cells contained well developed Golgi complex and abundant RER. In the experiment group of the visceral painful stimulation the results were as follows: There were more striking changes of parenchymal cells of anterior lobes in rats pituitary than those of the experimental group of the somatic painful stimulation. The cell bodies of ACTH, GH and PRL cells showed hypertrophy, the nucleus enlarged and nucleolus were relatively prominent. The dense granules decreased in number and the pale or empty visicles almost situated near the plasmic membrane in ACTH cells. In the GH cells large vacuoles were distributed throughout the cytoplasma. The number and density of the secretory granules decreased. In PRL cells the Golgi complex were extended, the dictyosomes increased, in addition, RER, polyribosomes and mitochandria were more aboundant than that in these normal cells. These cytological changes suggested secretory and synthetic activity of these cells were enhanced. At last, pain in relation to several hormones such as ?-endorphin, ?-LPH, ACTH and ?-MSH etc. were discussed by the author.
ABSTRACT
Pain just tolerable was caused intermittently by electric stimulation on rat's tail or by distented baloon in stomach for 30 min. The changes of intermediate lobe are summarized as follows: (1) The cell body, nucleus or nucleolus was hypertrophied. The number of rough endoplasmic reticulum, particularly of its vesicular cisternae was increased. Golgi complex enlarged. The ribosome multiplied. All of these change indicated that the activity of cells was increased and the rate of synthesis enhanced. (2) Both the secretory granules shifted to the periphery of cell body and some of them showed pale density. These changes were interpreted as increase of seretory activity. (3) The mitochondria became enlarged and the crsitae in them were changed into tubular form. This suggested an acceleration of cell metabolism, which might provide a large amount of energy. (4) There was no degenerative change. (5) The cytological changes caused by visceral pain were simillar with, but more striking than, those caused by somatic pain. (6) The secretions were increased, among which ?-endorphin and ACTH etc were known to be closely related to analgesia.