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Background: Babesiosis is a socioeconomically typical tick-borne infectious disease. Babesia infections in small mammals and ticks were reported in at least five provinces in China. However, the host range and geographical distribution of this parasite in Fujian province was unclear. The aim of this work was to investigate the prevalence and genetic diversity of Babesia in Fujian province, Southeast China, between 2015 and 2020.Methods: Blood samples of rodents were collected from 26 various types of surveillance sites accross Fujian province. Genomic DNA was extracted for screening the Babesia infection via PCR amplification based on 18S rRNA. The prevalences of Babesia were compared by the Chi-square test or Fisher exact test. Phylogenetic tree was constructed using MEGA 5.0 by gene sequence alignment. DNA samples of 316 Capra Caprinae, 85 Bos Bovine, 56 Canis lupus familiaris, 27 Lepus sinensis and 18 Sus scrofa domesticus were also collected in this survey.Results: A total of 1123 rodents were trapped and tested. Babesia infection was confirmed in 3.83% (43/1123) of the rodents and in 1.20% (6/502) of other trapped mammals. Multivariate logistic 2 regression analysis revealed that irrigated cropland, reeds, shrub and hills were risk factors for Babesia infection. The infection rate of S.scrofa domesticus, C.lupus familiaris and C.Caprinae were determine to be 5.56%, 1.79% and 1.27%, and no infection were found in B.Bovine and L.sinensis. There was a significant difference in the prevalence of Babesia between rodents and other host animals.Conclusions: The results indicated that there exits a broad geographical distribution and genetic diversity of Babesia in Southeast China. This study suggested that the mammals, especially wild rodents were the main natural hosts of Babesia in Fujian. Our research provided new insights on the exposure risk of Babesia in humans and animals, laying a solid foundation for the development of babesiosis prevention and control measures.

To explore the status of Babesia infection in rodents and the genetic characteristics of Babesia spp. in Fujian Province.Rodents were captured by the night trapping method in Shaowu, Qingliu, Shunchang, Yong’an, Changle and Youxi during 2014-2015. The rodent species was identified, and information on the time and place of capture, species and sex of rodents was recorded. Blood samples was collected, in which the fragment of 18S rRNA gene of Babesia spp. was amplified by PCR. The PCR products were sequenced and the phylogenetic tree was constructed for homology analysis. Data on positive rate were analyzed with Chi-square or Fisher exact test.Two hundred and nine rats were captured, comprising of 71 domestic and 138 wild rats. The overall positive rate was 9.6%(20/209). The positive rate in domestic rats was 2.8%(2/71), including one Rattus norvebicus and one Rattus flavipectus. The positive rate in wild rats was 13.0%(18/138), including 13 Bandicota indica, one Rattus losea, 2 Rattus confucianus and 2 Rattus fulvescens. The positive rate was significantly higher in wild rats than in domestic rats (P0.05). The Youxi region had the highest positive rate(14.9%,13/87), followed by Yong’an(13.6%, 3/22), and no positive rat was found in Qingliu. The positive rate in the male rats was 7.9%(9/114), and that in the females was 11.6%(11/95). The positive rate was highest in adult rats (10.4%,18/173), followed by young ones (6.3%,2/32). No positive rat was found in old rats. There was no significant difference in positive rate among different regions, between male and female rats, or among different ages (P0.05). The sequences of PCR products had a 100% homology. The BLAST results revealed the species to be Babesia microti. The phylogenetic tree showed that the sample sequence was the most homologous with Babesia microti from Zhejiang Province(GenBank Accession No: JQ609305).There occurs Babesia microti infection in rats in part areas of Fujian Province. The positive rate was higher in wild rats than in domestic rats.

In rodent models of traumatic brain injury (TBI), both Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNFα) levels increase early after injury to return later to basal levels. We have developed and characterized a rat mild fluid percussion model of TBI (mLFP injury) that results in righting reflex response times (RRRTs) that are less than those characteristic of moderate to severe LFP injury and yet increase IL-1α/β and TNFα levels. Here we report that blockade of IL-1α/β and TNFα binding to IL-1R and TNFR1, respectively, reduced neuropathology in parietal cortex, hippocampus, and thalamus and improved outcome. IL-1β binding to the type I IL-1 receptor (IL-1R1) can be blocked by a recombinant form of the endogenous IL-1R antagonist IL-1Ra (Kineret). TNFα binding to the TNF receptor (TNFR) can be blocked by the recombinant fusion protein etanercept, made up of a TNFR2 peptide fused to an Fc portion of human IgG1. There was no benefit from the combined blockades compared with individual blockades or after repeated treatments for 11 days after injury compared with one treatment at 1 hr after injury, when measured at 6 hr or 18 days, based on changes in neuropathology. There was also no further enhancement of blockade benefits after 18 days. Given that both Kineret and etanercept given singly or in combination showed similar beneficial effects and that TNFα also has a gliotransmitter role regulating AMPA receptor traffic, thus confounding effects of a TNFα blockade, we chose to focus on a single treatment with Kineret.

One new flavone, 5,7-dihydroxy-8,2',3',6'-tetramethoxyflavone (1), and one new flavonol, 5,7,6'-trihydroxy-2'-methoxyflavonol (2), were isolated from the roots of Scutellaria baicalensis, and their structures were elucidated on the basis of extensive spectroscopic evidences, especially 1D and 2D NMR and HR-ESI-MS experiments. The structure features of these new compounds lie in the substitution at both C-2' and C-6' positions of B-ring by hydroxyl or methoxyl groups.

One of the criteria defining mild traumatic brain injury (mTBI) in humans is a loss of consciousness lasting for less than 30 min. mTBI can result in long-term impairment of cognition and behavior. In rats, the length of time it takes a rat to right itself after injury is considered to be an analog for human return to consciousness. This study characterized a rat mild brain blast injury (mBBI) model defined by a righting response reflex time (RRRT) of more than 4 min but less than 10 min. Assessments of motor coordination relying on beam-balance and foot-fault assays and reference memory showed significant impairment in animals exposed to mBBI. This study's hypothesis is that there are inflammatory outcomes to mTBI over time that cause its deleterious effects. For example, mBBI significantly increased brain levels of interleukin (IL)−1β and tumor necrosis factor-α (TNFα) protein. There were significant inflammatory responses in the cortex, hippocampus, thalamus, and amygdala 6 hr after mBBI, as evidenced by increased levels of the inflammatory markers associated with activation of microglia and macrophages, ionized calcium binding adaptor 1 (IBA1), impairment of the blood–brain barrier, and significant neuronal losses. There were significant increases in phosphorylated Tau (p-Tau) levels, a putative precursor to the development of neuroencephalopathy, as early as 6 hr after mBBI in the cortex and the hippocampus but not in the thalamus or the amygdala. There was an apparent correlation between RRRTs and p-Tau protein levels but not IBA1. These results suggest potential therapies for mild blast injuries via blockade of the IL-1β and TNFα receptors. © 2014 Wiley Periodicals, Inc.

This document analyzes the limit of duty circle in a conventional two-transistor forward DC/DC converter which is disadvantageous to efficiency. To solve this problem, the resonant reset technology is introduced to the converter. The enhanced two-transistor forward DC/DC converter is able to increase its duty circle exceeding 50% and the resonance voltage is constant which is propitious to heighten the efficiency. Simulation is implemented for the purpose of validating the feasibility.

This document proposes a novel two-transistor forward DC/DC converter with resonance reset. It is recommended because the duty circle is able to exceed 50% and the efficiency of the converter is higher which is superior to the conventional. The principle of operation of the enhanced configuration is illustrated and some simulation is employed to prove its practicability and superiority. Finally a 80V~140V input, 12V/8.3A output DC/DC converter prototype is founded for verification.

This document explains and demonstrates the composing of power losses of MOSFETs in DC/DC converters which are based on Single-ended Forward topology and Resonant Reset technology and how to calculate the losses by equations. For the purpose of validating the correctness of these equations, two different MOSFETs are chosen and uesed in the same DC/DC converter. Furthermore, some practical method is presented in choosing the most suited MOSFETs for certain DC/DC converters.

Chronic central neuropathic pain after central nervous system injuries remains refractory to therapeutic interventions. A novel approach would be to target key intracellular signaling proteins that are known to contribute to continued activation by phosphorylation of kinases, transcription factors, and/or receptors that contribute to changes in membrane excitability. We demonstrate that one signaling kinase, calcium/calmodulin-dependent kinase II (CaMKII), is critical in maintaining aberrant dorsal horn neuron hyperexcitability in the neuropathic pain condition after spinal cord injury (SCI). After contusion SCI at spinal level T10, activated CaMKII (phosphorylated, pCaMKII) expression is significantly upregulated in the T7/8 spinal dorsal horn in neurons, but not glial cells, and in oligodendrocytes in the dorsal column in the same rats that displayed at-level mechanical allodynia. Furthermore, identified spinothalamic neurons demonstrated significant increases of pCaMKII after SCI compared to sham-treated control animals. However, neither astrocytes nor microglia showed pCaMKII expression in either sham-treated or SCI rats. To demonstrate causality, treatment of SCI rats with KN-93, which prevents CaMKII activation, significantly attenuated at-level mechanical allodynia and aberrant wide dynamic range neuronal activity evoked by brush, pressure, and pinch stimuli and a graded series of von Frey stimuli, respectively. Persistent CaMKII activation contributes to chronic central neuropathic pain by mechanisms that involve maintained hyperexcitability of wide dynamic range dorsal horn neurons. Furthermore, targeting key signaling proteins is a novel, useful therapeutic strategy for treating chronic central neuropathic pain.

Elevation of extracellular glutamate contributes to cell death and functional impairments generated by spinal cord injury (SCI), in part through the activation of the neurotoxic cytokine interleukin-1beta (IL-1beta). This study examines the participation of IL-1beta and its regulation by the endogenous interleukin-1 receptor antagonist (IL-1ra) in glutamate toxicity following SCI. Glutamate, glutamatergic agonists and SCI had similar effects on levels of IL-1beta and IL-1ra. Following spinal cord contusion or exposure to elevated glutamate, concentrations of IL-1beta first increased as IL-1ra decreased, and both then changed in the opposite directions. Applying the glutamate agonists NMDA and S-AMPA to the spinal cord caused changes in IL-1beta and IL-1ra levels very similar to those produced by contusion and glutamate. The glutamate antagonists MK801 and NBQX blocked the glutamate-induced changes in IL-1beta and IL-1ra levels. Administering IL-1beta elevated IL-1ra, and administering IL-1ra depressed IL-1beta levels. Infusing IL-beta into the spinal cord impaired locomotion, and infusing IL-1ra improved recovery from glutamate-induced motor impairments. We hypothesize that elevating IL-1ra opposes the damage caused by IL-1beta in SCI by reducing IL-1beta levels as well as by blocking binding of IL-1beta to its receptor. Our results demonstrate that IL-1beta contributes to glutamate damage following SCI; blocking IL-1beta may usefully counteract glutamate toxicity.

A neuroprotective factor is shown to be present in mammalian serum. This factor is identified by Western blotting to be serum albumin. The serum factor and albumin both protected cultured spinal cord neurons against the toxicity of glutamate. The inability of K252a, a blocker of the high affinity tyrosine kinase receptor for members of the nerve growth factor family, to block the neuroprotective effect of the serum factor established that the serum factor is not a member of the nerve growth factor family. Post-injury injection of albumin intravenously or into the site of injury immediately after injury both improved significantly locomotor function according to Basso-Beattie-Bresnahan assessment and spontaneous locomotor activity recorded with a photobeam activity system. Albumin has multiple mechanisms whereby it may be neuroprotective, and it is a potentially useful agent for treating neurotraumas.

Rats given moderate spinal cord injury (SCI) display increases in the expression of the activated form of the transcription factor cyclic AMP responsive element binding protein (CREB) in spinal segments of dermatomes corresponding to permanent mechanical allodynia, a model of chronic central neuropathic pain (CNP; (Crown, E.D., Ye, Z., Johnson, K.M., Xu, G.Y., McAdoo, D.J., Westlund, K.N., Hulsebosch, C.E., 2005. Upregulation of the phosphorylated form of CREB in spinothalamic tract cells following spinal cord injury: relation to central neuropathic pain. Neurosci. Lett. 384, 139-144)). Given that not all rats that receive moderate SCI develop CNP, the current study was designed to further analyze changes in persistent CREB activation and in the activation state of upstream intracellular signaling cascades (e.g., mitogen-activated protein kinases [MAPKs]) in populations of rats that receive SCI and weeks later develop CNP and rats that receive SCI but do not develop CNP. The results indicate that activated kinases such as pERK 1/2, p-p38 MAPK, but not pJNK, are upregulated in injured rats that develop CNP as compared to injured rats that fail to develop CNP. In addition, the current results replicated our previous finding that activated CREB is upregulated following SCI, however, only in SCI rats that developed CNP. Taken together, these results indicate that activation of intracellular signaling cascades traditionally associated with long-term potentiation and memory is associated with the expression of chronic CNP following SCI.

Spinal cord injury (SCI) often leads to the generation of chronic intractable neuropathic pain. The mechanisms that lead to chronic central neuropathic pain (CNP) following SCI are not well understood, resulting in ineffective treatments for pain relief. Studies have demonstrated persistent hyperexcitability of dorsal horn neurons which may provide a substrate for CNP. We propose a number of similarities between CNP mechanisms and mechanisms that occur in long-term potentiation, in which hippocampal neurons are hyperexcitable. One biochemical similarity may be activation of the transcription factor, cyclic AMP response element-binding protein (CREB), via phosphorylation (pCREB). The current study was designed to examine whether tactile allodynia that develops in segments rostral to SCI (at-level pain) correlates with an increase in CREB phosphorylation in specific neurons known to be involved in allodynia, the spinothalamic tract (STT) cells. This study determined that, in animals experiencing at-level allodynia 35 days after SCI, pCREB was upregulated in the spinal cord segment rostral to the injury. In addition, pCREB was found to be upregulated specifically in STT cells in the rostral segment 35 days after SCI. These findings suggest one mechanism of maintained central neuropathic pain following SCI involves persistent upregulation of pCREB expression within STT cells.

In vivo experiments addressing the role of released glutamate in damage caused by neurotrauma seldom administer glutamate itself because it usually produces relatively little damage when administered into central nervous system (CNS) tissue in vivo. However, because of recent observations that glutamate administered into the spinal cord at the levels attained following spinal cord injury (SCI) kills neurons and oligodendrocytes, we tested the effects of administering glutamate at those concentrations on locomotor function. The Basso-Beattie-Bresnahan (BBB) test and activity box measures demonstrated that those glutamate concentrations produce lasting functional impairments. Several parameters provided by the activity box provided sensitive measures of the degree of post-SCI impairment, demonstrating their substantial potential for evaluating outcomes of SCI. Results obtained also enhance evidence that glutamate toxicity contributes to secondary damage following SCI and suggest that damage to white matter is an important contributor to such damage.

Spinal cord injury (SCI) induces neuronal death, including apoptosis, which is completed within 24 hr at and around the impact site. We identified early proapoptotic transcriptional changes, including upregulation of proapoptotic Bax and downregulation of antiapoptotic Bcl-xL, Bcl-2, and Bcl-w, using Affymetrix DNA microarrays. Because Bcl-xL is the most robustly expressed antiapoptotic Bcl-2 molecule in adult central nervous system, we decided to characterize better the effect of SCI on Bcl-xL expression. We found Bcl-xL expressed robustly throughout uninjured spinal cord in both neurons and glia cells. We also found Bcl-xL localized in different cellular compartments: cytoplasmic, mitochondrial, and nuclear. Bcl-xL protein levels decreased in the cytoplasm and mitochondria 2 hr after SCI and persisted for 24 hr. To test the contribution of proapoptotic decreases in Bcl-xL to neuronal death, we augmented endogenous Bcl-xL levels by administering Bcl-xL fusion protein (Bcl-xL FP) into injured spinal cords. Bcl-xL FP significantly increased neuronal survival, suggesting that SCI-induced changes in Bcl-xL contribute considerably to neuronal death. Because Bcl-xL FP increases survival of dorsal horn neurons and ventral horn motoneurons, it could become clinically relevant in preserving sensory and motor functions after SCI.

Spinal cord injury (SCI)-induced neurodegeneration leads to irreversible and devastating motor and sensory dysfunction. Post-traumatic outcomes are determined by events occurring during the first 24 hours after SCI. An increase in extracellular glutamate concentration to neurotoxic levels is one of the earliest events after SCI. We used Affymetrix DNA oligonucleotide microarrays (with 1,322 DNA probes) analysis to measure gene expression in order to test the hypothesis that SCI-induced N-methyl-D-aspartate (NMDA) receptor activation triggers significant postinjury transcriptional changes. Here we report that SCI, 1 hour after trauma, induced change in mRNA levels of 165 genes and expression sequence tags (ESTs). SCI affected mRNA levels of those genes that regulate predominantly transcription factors, inflammation, cell survival, and membrane excitability. We also report that NMDA receptor inhibition (with -(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]-cyclohepten-5,10-imine hydrogen maleate [MK-801]) reversed the effect of SCI on about 50% of the SCI-affected mRNAs. Especially interesting is the finding that NMDA receptor activation participates in the up-regulation of inflammatory factors. Therefore, SCI-induced NMDA receptor activation is one of the dominant, early signals after trauma that leads to changes in mRNA levels of a number of genes relevant to recovery processes. The majority of MK-801 effects on the SCI-induced mRNA changes reported here are novel. Additionally, we found that the MK-801 treatment also changed the mRNA levels of 168 genes and ESTs that had not been affected by SCI alone, and that some of their gene products could have harmful effects on SCI outcome.

Spinal cord injury (SCI) leads to an increase in extracellular excitatory amino acid (EAA) concentrations, resulting in glutamate receptor-mediated excitotoxicity and central sensitization. To test contributions of group I metabotropic glutamate receptors (mGluRs) in SCI induced release of glutamate and in behavioral outcomes of central sensitization following injury, we administered 1-aminoindan-1,5-dicarboxylic acid (AIDA; 0.1 nmol intraspinally), a potent group I mGluR antagonist, to rats immediately after spinal cord contusion injury. EAAs were collected by microdialysis and quantified using HPLC. AIDA significantly decreased extracellular glutamate but not aspartate concentrations and significantly attenuated the development of mechanical but not thermal allodynia. These results suggest mGluRs play an important role in injury-induced EAA release and in central sensitization following SCI.

Damage caused by administering glutamate into the spinal cord was characterized histologically. Glutamate destroyed neurons for several hundred micrometers around the administering microdialysis fiber. At 24 h after treatment, significant (P=0.036) loss of neurons was observed (75%) relative to control (47%) near the fiber when glutamate was administered for 1 h at a concentration outside the fiber approximating the maximum glutamate released upon spinal cord injury. Significant loss of neurons (P=0.006, 0.022) was also caused by administering a combination of glutamate at about its average concentration released upon injury over the 1 h period of administration in combination with the maximum aspartate concentration released upon injury. This work provides a direct demonstration that the concentrations of excitatory amino acids released upon spinal cord injury are neurotoxic. The destruction of neurons by exposure to excitatory amino acids when there is also substantial loss of neurons simply from the presence of the microdialysis fiber may reflect sensitization of neurons to excitotoxicity by stress.

The following issues are further addressed: (1) Is there considerable leakage of amino acids from the circulation into the space around microdialysis probes, or are amino acid concentrations naturally much higher in the interstitial space than is generally thought? (2) Do observed high interstitial concentrations or depletion of substances in the intracellular space by microdialysis affect release measurements upon spinal cord injury? Amino acid concentrations around microdialysis fibres in the spinal cord of rats were found to approach those in the circulation and to be much higher than interstitial concentrations previously estimated in the CNS. However, much lower concentrations of amino acids were derived in the hippocampus by analogous experiments. Considerable Evans Blue/albumin leaked from the circulation into the interstitial space in the spinal cord immediately after fibre insertion. However, this movement diminished considerably by 4 h later, demonstrating substantial resealing of the blood-brain barrier, at least to large molecules. There is either substantial damage-induced movement of amino acids from the circulation into the dialysis zone after insertion of a microdialysis probe, or there is much less impediment to movement of amino acids across the blood-brain barrier in the spinal cord than in the brain. At low flow rates through the fibre, adding concentrations of amino acids to the inside of the fibre equal to the concentrations around the fibre to prevent their depletion by removal through the microdialysis fibre did not affect increases in concentrations of amino acids in microdialysates following injury. Thus the high concentrations of amino acids present around microdialysis fibres following their insertion do not seem to disturb measurements of amino acid release upon spinal cord injury.

Under global cerebral ischemia, the effect of different brain temperature on cerebral ischemic injury was studied. Male Sprague-Dawley rats were divided into normothermic (37-38°C) ischemia, mild hypothermic (31-32°C) ischemia, hyperthermic (41-42°C) ischemia and sham-operated groups. Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model and brain temperature was maintained at defined level for 60 min after 20-min ischemia. The expression of c-fos protein and the levels of malondialdehyde (MDA) and lactate in brain regions were detected by immunochemistry and spectrophotometrical methods, respectively. C-fos positive neurons were found in the hippocampus and cerebral cortex after cerebral ischemia reperfusion. Mild hypothermia increased the expression of c-fos protein in both areas, whereas hyperthermia decreased the expression of c-fos protein in the hippocampus at 24 h reperfusion, and the cerebral cortex at 48 h reperfusion when compared to normothermic conditions. In normothermic, mild hypothermic and hyperthermic ischemia groups, the levels of MDA and lactate in brain tissue were increased at 24, 48 and 72 h reperfusion following 20-min ischemia as compared with the sham-operated group (P

To explore the roles of NF-κB in factor VIIa-induced proliferation and migration of a colon cancer cell line (SW620) in vitro and its possible mechanism.The expression levels of NF-κB (p65), inhibitory protein of NF-κB (IκB-α), caspase-7, interleukin 8 (IL-8) and tissue factor (TF) in SW620 cells treated with factor VIIa, PDTC (an inhibitor of NF-κB) and other factors were measured by Western-blotting and real-time PCR. Proliferation and migration of the cells were analyzed by flow cytometry and Transwell assay, respectively.Factor VIIa down-regulated the IκB-α level in SW620 cells and increased the intranuclear level of NF-κB. Those effects of factor VIIa were blocked by anti-TF or anti-PAR2 antibodies. The effects of factor VIIa on proliferation and migration of SW620 cells, expression of IL-8, TF as well as caspase-7, were interfered by PDTC (the inhibitor of NF-κB).TF/VIIa complex activates NF-κB pathway via PAR2, thereby up-regulates IL-8 and down-regulates caspase-7 expression in SW620 cells, finally promotes proliferation and migration of colon cancer cells. In addition, TF/VIIa/PAR2/NF-κB pathway also upregulates TF expression, thus to create a positive feedback loop of TF/VIIa/PAR2/NF-κB/TF.

To investigate whether tumor necrosis factor receptor-associated factor 6 (TRAF6) is involved in anti-β2GPI/β2GPI-induced tissue factor (TF) expression on THP-1 cells.The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with anti-β2GPI/β2GPI complex. And then the TF expression on THP-1 cells was detected by real-time quatitative PCR (RT-qPCR) and TF activity kit. TRAF6 mRNA and its protein expression were investigated by RT-qPCR and Western blotting, respectively. The proteasome inhibitor, MG-132, was used for inhibitory assays, in order to demonstrate the effect of anti-β2GPI/β2GPI complex on THP-1 cells.The TF expression (both mRNA and activity) on THP-1 cells was significantly up-regulated with the treatment of anti-β2GPI/β2GPI complex (100 mg/L), compared with untreated cells(P0.05). The TRAF6 mRNA and protein levels in THP-1 cells were also significantly increased with the treatment of anti-β2GPI/β2GPI complex. The expression of TRAF6 was shown in a time-dependent manner, with the maximal level at 15 minutes (mRNA) and 30 minutes (protein) respectively. All the stimulating effects of anti-β2GPI/β2GPI complex (100 mg/L) on THP-1 cells were inhibited by MG-132 (5 μmol/L).TRAF6 is up-regulated and contributes to TF expression on THP-1 cells induced with anti-β2GPI/β2GPI complex.

Released excitatory amino acids contribute significantly to secondary damage following spinal cord injury. Reversal of normal transport due to cell membrane depolarization may contribute to this release. We tested this by administering dihydrokainic acid (DHK), a non-transported glutamate uptake blocker, into the rat spinal cord by microdialysis in association with contusion spinal cord injury. Glutamate release in response to injury was reduced by 34% (P

To establish recombinant outer membrane lipoprotein LipL32-based antibody detection assays in identifying leptospirosis.Recombinant leptospiral outer membrane protein LipL32 was obtained by genetic engineering method. This purified protein was used in the indirect and sandwich ELISA assays to test the antibodies in sera of human beings and rats, and the results were compared with those obtained by microscopy agglutination test (MAT) and imported ELISA kit.When the acute and convalescent phase specimens from 9 leptospiral patients were tested, the detected rates of three ELISAs were similar to the MAT. Among the 45 probable cases which MAT showed positive, 32 (71.11%) samples were positive by r32-I-ELISA, 36 (80.00%) by r32-S-ELISA, while 28.89% (13/45) samples were positive and 55.56% (25/45) were suspicious by D.A.I-ELISA. The specificity of r32-I-ELISA and r32-S-ELISA were 97.10% (67/69) for 69 specimens. 43 healthy specimens were negative by both r32-I-ELISA and r32-S-ELISA, 14 healthy specimens were negative by D.A.I-ELISA. Among 16 non-leptospirosis patients, two specimens were positive by r32-I-ELISA and r32-S-ELISA, D.A.I-ELISA and identified one positive specimen, while 12 specimens were suspicious by D.A.I-ELISA. For 10 syphilis specimens, data obtained through three ELISAs were in consistent with that by MAT. A sandwiched ELISA, using rLipL32 protein as the antigen was developed to detect rat sera. Considering MAT as standard test, the sensitivity and specificity were 86.75% (131/151), 99.19% (122/123) respectively with coincidence rate as 92.34% (253/274).The recombinant protein LipL32 had high immunoreactivity and could be used as an antigen for the detection of pathogenic leptospirosis. In summary, the novel sandwiched ELISA with rLipL32 showed similar sensitivity and specificity to that of MAT in the antibody detection of rat leptospirosis. It was suitable for large scales field sero-epidemiological studies.

The toxicity of released glutamate contributes substantially to secondary cell death following spinal cord injury (SCI). In this work, the extent and time courses of glutamate-induced losses of neurons and oligodendrocytes are established. Glutamate was administered into the spinal cords of anesthetized rats at approximately the concentration and duration of its release following SCI. Cells in normal tissue, in tissue exposed to artificial cerebrospinal fluid and in tissue exposed to glutamate were counted on a confocal system in control animals and from 6 h to 28 days after treatment to assess cell losses. Oligodendrocytes were identified by staining with antibody CC-1 and neurons by immunostaining for Neuronal Nuclei (NeuN) or Neurofilament H. The density of oligodendrocytes declined precipitously in the first 6 h after exposure to glutamate, and then relatively little from 24 h to 28 days post-exposure. Similarly, neuron densities first declined rapidly, but at a decreasing rate, from 0 h to 72 h post-glutamate exposure and did not change significantly from 72 h to 28 days thereafter. The nuclei of many cells strongly and specifically stained for activated caspase-3, an indicator of apoptosis, in response to exposure to glutamate. Caspase-3 was localized to the nucleus and may participate in apoptotic cell death. However, persistence of caspase-3 staining for at least a week after exposure to glutamate during little to no loss of oligodendrocytes and neurons demonstrates that elevation of caspase-3 does not necessarily lead to rapid cell death. Beyond about 48 h after exposure to glutamate, locomotor function began to recover while cell numbers stabilized or declined slowly, demonstrating that functional recovery in the experiments presented involves processes other than replacement of oligodendrocytes and/or neurons.

To investigate the efficacy of 10 mg or 20 mg atorvastatin + long acting antihypertensive in carotid intima-medial thickness (IMT).151 patients of Han nationality in South China with mild hypertensive were randomly divided into 3 groups: atorvastatin 10 mg group (n = 50) receive 10 mg atorvastatin and amlodipine + benazepril; atorvastatin 20 mg group (n = 61) receive 20 mg atorvastatin and amlodipine + benazepril; the control group (n = 40) receive amlodipine + benazepril. The patients were detected IMT, vascular function, lipids and inflammatory factor in pretherapy and every 3 months.atorvastatin 10 mg or 20 mg groups have significantly change contrast to control group: (1) IMT was decreased (P0.01). (2) Deltadia-P% and Deltadia-N% were increased (P0.01). (3) LDL-C level was decreased by 30% in a atorvastatin 10 mg group and 40.48% in 20 mg group respectively (P0.01).Atorvastatin delays the development of atherosclerosis in hypertensive patients, improves endothelial function, and strengthens the effect of lipid-lowering.

Central neuropathic pain (CNP) is an important problem following spinal cord injury (SCI), because it severely affects the quality of life of SCI patients. As in the patient population, the majority of rats develop significant allodynia (CNP rats) after moderate SCI. However, about 10% of SCI rats do not develop allodynia, or develop significantly less allodynia than CNP rats (non-CNP rats). To identify transcriptional changes underlying CNP development after SCI, we used Affymetrix DNA microarrays and RNAs extracted from the spinal cords of CNP and non-CNP rats. DNA microarry analysis showed significantly increased expression of a number of genes associated with inflammation and astrocytic activation in the spinal cords of rats that developed CNP. For example, mRNA levels of glial fibrilary acidic protein (GFAP) and Aquaporin 4 (AQP4) significantly increased in CNP rats. We also found that GFAP, S100beta and AQP4 protein elevation persisted for at least 9 months throughout contused spinal cords, consistent with the chronic nature of CNP. Thus, we hypothesize that CNP development results, in part, from dysfunctional, chronically "over-activated" astrocytes. Although, it has been shown that activated astrocytes are associated with peripheral neuropathic pain, this has not previously been demonstrated in CNP after SCI.

The authors previously deduced a formula for calculating the steal index (SI) for evaluation of the steal phenomena during the development of the collateral circulation resulting from unilateral carotid artery occlusion or stenosis. In this study the authors examine the application of SI in evaluating the steal phenomena of cerebral blood flow in cases of intracranial arteriovenous malformation (AVM).The clinical data of 16 cases of AVM confirmed by both digital subtraction angiography (DSA) and transcranial Doppler (TCD) ultrasound were analyzed retrospectively. The ratio of the mean blood velocity in the direct or indirect feeding artery (Vmfv) for the area containing AVM to the mean blood velocity in the neighboring feeding artery (Vmna), Vmfv/Vmna, was compared with SI of the corresponding arteries (SI=1-PI(1)/PI(2), where PI(1) is the pulsatility index of the feeding artery and PI the pulsatility index of the neighboring feeding artery), and the correlation between SI and the size of AVM was analyzed statistically.The Vmfv/Vmna of (2)direct and indirect feeding arteries for AVM both exhibited positive correlation with SI (r=0.62, P0.05, n=16; r=0.53, P0.01, n=27), and SI appeared to be in positive correlation with the size of AVM, but which failed to be supported by statistical analysis (r=0.48, P0.05, n=12).The sizes, types and development of the vascular bed of AVM can be evaluatea by analysis of SI derived from TCD in combination with DSA.

To explore multi-drug resistance (MDR) of bladder cancer for the intravesical instillation.Using immunohistochemical staining, in 44-case human bladder cancer cells, the expressions of P-glycoprotein (P-gp), glutathione S-transferase (GST-pi) and topoisomerase (TOPO-II), were detected to find out the resistance to drugs.P-gp had a higher expression in 54.5% cases. GST-pi had no or a lower expression in 65.9% cases. TOPO-II had a higher expression in 29.5% but a lower expression in 65.9% cases.Detecting the factors of MDR in bladder cancer cells could help to choose drugs for intravesical chemotherapy.

The release of reactive oxygen species (ROS) by neutrophils, which infiltrate the region of damage following spinal cord injury (SCI), was investigated to determine if such release is significant following spinal cord injury. The relationship of extracellular levels of hydroxyl radicals and hydrogen peroxide obtained by microdialysis sampling and oxidized protein levels in tissue to neutrophil infiltration following spinal cord injury was examined. Neither of the reactive oxygen species were elevated in the site of spinal cord injury relative to their concentrations in normal tissue at a time (24 h) when the numbers of neutrophils were maximum in the site of injury. Surprisingly, ablation with a neutrophil antiserum actually increased the level of oxidized proteins in Western blots. Thus, our findings are (1) that neutrophils, which infiltrate the site of damage following a spinal cord injury, do not release detectable quantities of reactive oxygen species; and (2) that the presence of neutrophils reduces the concentrations of oxidized proteins in the site of spinal cord injury. Therefore, release of reactive oxygen species by neutrophils does not contribute significantly to secondary damage following spinal cord injury. Reduced levels of oxidized proteins in the presence of neutrophils may reflect removal of damaged tissue by neutrophils.

We investigated in vivo in rats whether sufficient glutamate is released following spinal cord injury (SCI) to kill oligodendrocytes. Microdialysis sampling was used to establish the level of glutamate released (550 +/- 80 microM) in the white matter during SCI. This glutamate concentration was administered into the spinal cords of other rats and the densities of oligodendrocytes remaining 24 and 72 h later determined by counting cells immunostained with the oligodendrocyte marker CC-1. Administration of ACSF, 4.0 mM glutamate (estimated resulting tissue exposure 500 microM) and 10.0 mM glutamate by microdialysis reduced oligodendrocyte density 22%, 57%, and 74%, respectively, relative to normal at 24 h post-exposure. Therefore, sufficient glutamate is released following SCI to damage white matter. Oligodendrocyte densities near the fiber track were not significantly different at 72 h from 24 h post-exposure, so most glutamate-induced oligodendrocyte death occurs within 24 h after exposure. Injecting the AMPA/kainate receptor blocker NBQX into the spinal cord during glutamate administration reduced the glutamate-induced decrease in oligodendrocyte density, evidence for AMPA/kainate receptor involvement in glutamate-induced oligodendrocyte death. This work directly demonstrates in vivo that following SCI glutamate reaches concentrations toxic to white matter and that AMPA/kainate receptors mediate this glutamate toxicity to oligodendrocytes.

To study the calculation of cerebral blood flow loss ratio after cerebral artery stenosis using the data obtained from transcranial Doppler sonography (TCD), by deducing regional cerebral blood flow loss index (rCBFLI).The data of TCD performed in 31 cases of unilateral stenosis in the middle cerebral artery (MCA) were collected and analyzed for deducing the formula for calculating the MCA stenosis index (STI): STI=1 (Vm(0)/Vm(1)) x (PI(1)/PI(0)), where Vm(0) is the mean normal velocity of MCA blood flow (derived from the measurements in 908 normal subjects at varied ages receiving TCD) and Vm(1) the mean velocity at the stenosed MCA, and PI(0) and PI(1) stand for the pulsatility index before and after stenosis. The ratio of PI(1) to PI(0) is likely to be equivalent to the ratio of Q(1) to Q(0) (Q(0) and Q(1) represent the volume of blood flow in the MCA in normal condition and after stenosis, respectively), thus the equivalence can be extended as rCBFLI=(1 Q(1)/Q(0)) x 100%=(1 PI(1)/PI(0)) x 100%. rCBFLI after MCA stenosis was calculated in 31 cases accordingly, and by means of correlation analysis, we found positive correlation between rCBFLI and Vm(1) (r=0.76, P0.001) and between Vm(1) and STI (r=0.85, P0.001). In the same way, rCBFLI of 55 sides with MCA stenosis in 43 cases were analyzed, and positive correlation between rCBFLI and Vm (r=0.76, P0.001) and between rCBFLI and STI (r=0.83, P0.001) was found.rCBFLI can be used to evaluate the degree of regional cerebral blood flow loss due to artery stenosis, and the combination of rCBFLI and STI may provide an insight into the changes of cerebral hemodynamics in this pathologic condition.

After contusion-derived spinal cord injury, (SCI) there is localized tissue disruption and energy failure that results in early necrosis and delayed apoptosis, events that contribute to chronic central pain in a majority of patients. We assessed the extent of contusion-induced apoptosis of neurons in a known central pain-signaling pathway, the spinothalamic tract (STT), which may be a contributor to SCI-induced pain. We observed the loss of STT cells and localized increase of DNA fragmentation and cytoplasmic histone-DNA complexes, which suggested potential apoptotic changes among STT neurons after SCI. We also showed SCI-associated changes in the expression of the antiapoptotic protein Bcl-xL, especially among STT cells, consistent with the hypothesis that Bcl-xL regulates the extent of apoptosis after SCI. Apoptosis in the injured spinal cord correlated well with prompt decreases in Bcl-xL protein levels and Bcl-xL/Bax protein ratios at the contusion site. We interpret these results as evidence that regulation of Bcl-xL may play a role in neural sparing after spinal injury and pain-signaling function.

Spinal cord injury (SCI) often results in abnormal pain syndromes in patients. We present a recently developed SCI mammalian model of chronic central pain in which the spinal cord is contused at T8 using the NYU impactor device (10-g rod, 2.0-mm diameter, 12.5-mm drop height), an injury which is characterized behaviorally as moderate. Recovery of locomotor function was assessed with an open field test and scored using the open field test scale (BBB scale). Somatosensory tests of paw withdrawal responses accompanied by supraspinal responses to both mechanical punctate (von Frey hairs) and nonpunctate (4 mm diameter blunt probe) as well as thermal (radiant heat) peripheral stimuli were performed. Comparisons at the level of the individual animal between precontusion and postcontusion responses indicated significant increases in reactions to low threshold punctate mechanical stimuli, non-punctate stimuli and thermal stimuli (p < 0.05). To demonstrate the validity of this model as a central pain model, gabapentin, an agent used clinically for central pain, was given i.p. at 10 or 30 mg/kg. Gabapentin treatment significantly and reversibly changed the responses, consistent with the attenuation of the abnormal sensory behavior, and the attenuated responses lasted for the duration of the drug effect (up to 6 h). These results support the use of the spinal contusion model in the study of chronic central pain after SCI.

The hypothesis that release of adenosine following spinal cord injury (SCI) may provide neuroprotective feedback is explored. Consistent with this hypothesis, substantial release of adenosine, estimated to reach 100 μM in the extracellular space, was detected by microdialysis sampling immediately following contusion SCI. There is also considerable release of excitatory amino acids following SCI. The latter was not affected by administration of the general adenosine receptor antagonist theophylline and the A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine, implying that the adenosine released following SCI does not significantly influence the release of neurotoxic amino acids. Administration of the concentration of glutamate released upon SCI into the spinal cord caused only about 1% as much release of adenosine as did injury, evidence that elevated excitatory amino acids do not elicit an appreciable fraction of the release of adenosine that follows SCI. Results obtained suggest that release of endogenous adenosine is not neuroprotective by blocking release of excitatory amino acids following SCI.

Release of amino acids, particularly the neurotoxin glutamate, in and around the site of an experimental spinal cord injury was characterized over time by microdialysis. Increases in amino acid concentrations caused by injury decline steeply and then slowly over distance from the impact area, becoming undetectable beyond about 5 mm from the injury epicenter. Diffusion profiles determined in the cord by administering amino acids through one microdialysis fiber and sampling them in a parallel fiber declined steeply with distance. Distant increases coincided temporally with those in the injury epicenter. We conclude that elevated amino acids more than about 1 mm into the periimpact zone are predominantly released in that region rather than diffusing into it from the trauma epicenter. In the outer areas of lesion development, glutamate does not appear to reach concentrations ordinarily toxic, and elevated concentrations do not persist nearly as long as the therapeutic window of NBQX in any part of the lesion. Therefore, the mechanisms whereby excitatory amino acid antagonists reduce the dimensions of injury lesions are unclear. However, sensitization of neurons following impact injury could be important in amino acid neurotoxicity.

Assorted microdialysis studies of the roles endogenous chemical agents may play in secondary damage upon spinal cord injury (SCI) are described. Issues addressed include the concentrations reached upon injury, mechanisms of release upon injury, and effects of drugs on injury-elicited increases in glutamate concentrations. An important question in identifying an agent of secondary damage upon central nervous system (CNS) trauma is not simply whether the substance is released upon injury, but whether it reaches harmful levels. To resolve this requires establishing the concentration attained and then determining whether administering that level damages neurons. To make microdialysis measurements of amino acids in the CNS more quantitative, we characterized the effects of insertion of a microdialysis fiber on leakage of glutamate from the circulation and explored the effects of depletion by microdialysis on release caused by SCI. Very high glutamate concentrations were found around the fiber for several hours after fiber insertion and 2 days later, and there was substantial leakage of alpha-aminoisobutyric acid from the circulation into the dialysis zone for several hours after fiber insertion. Glutamate concentrations reached upon SCI under nondepleting conditions were similar to those estimated earlier under depleting conditions. Mg2+ release was detectable when microdialysis probes were perfused with Mg2+-free fluid, but not when the concentrations in the perfusing fluid approximated those in the interstitial space. It is concluded (1) that insertion of microdialysis probes into CNS tissue can cause long-lasting leakage of amino acids from the circulation into the space around the fiber, (2) that this leakage can obscure concentration changes that otherwise occur, and (3) that depletion of substances in the fluid around the fiber may cause increases in concentration to be observed that do not normally happen. We also describe demonstrations that administration of methylprednisolone and dihydrokainic acid diminish increases in glutamate concentrations caused by SCI, showing that microdialysis can be used to explore effects of drugs on actions of damaging substances.

Changes of plasma cAMP and cGMP levels in chronic atrophic gastritis patients with spleen-qi deficiency

Analytic solution of the master equation using the exponential transition probability has been obtained previously in part III [J. Chem. Phys. 80, 2504 (1984)] in the form of an infinite series eigenfunction expansion for c(x,t), the population distribution. While the number of terms that effectively contribute to the sum is only one at equilibrium, it increases to infinity at time zero. Thus such eigenfunction expansion is not useful for describing the bulk properties [i.e., averages over c(x,t)] of the relaxing system at early times. It is nevertheless possible to solve the relaxation problem at early times by noting that the final (postcollision) energy distribution resulting from the nth collision is in fact the initial energy distribution for the next [(n+1)th] collision. It is shown that in this way simple analytical expressions can be obtained for various bulk properties of the relaxing system from the first collision onward—but not all the way to equilibrium—if the initial (at time zero) energy di...

If the internal energy of target molecules highly dispersed in a heat bath is monitored as a function of time, their one-photon laser excitation and subsequent bulk collisional relaxation yields information only about energy transfer in the bulk system (a macroscopic time-dependent property). This paper discusses three special cases when [Formula: see text], the average energy transferred in a collision (a microscopic property) can be deduced from bulk relaxation data without knowledge of the collisional transition probability: (i) initial excitation is a δ function; (ii) relaxation of bulk average energy follows simple exponential law; or (iii) it is linear in time. The implications of exponential relaxation is that [Formula: see text], which is the first moment of the collisional transition probability, is a linear function of internal energy. These conclusions are illustrated using available laser relaxation data on azulene, benzene, and hexafluorobenzene, and are compared with similar data on toluene and two cycloheptatrienes. Some inconsistencies are noted, the probable origin of which is discussed.