Qing-Dong Ling

A minimal stress model for the assessment of electroacupuncture analgesia in rats under halothane.

The use of anesthetics in acupuncture analgesia is controversial. We evaluate a steady‐state light anesthesia model to test whether minimal stress manipulation and reliable measurement of analgesia could be simultaneously achieved during electroacupuncture (EA) in animals.

A tetrameric glycoprotein Ib-binding protein, agglucetin, from Formosan pit viper: structure and interaction with human platelets

Agglucetin, a tetrameric agglutination inducer from the Formosan pit viper, has been identified as a platelet membrane glycoprotein (GP) Ib agonist and directly agglutinated fixed-platelets in the absence of von Willebrand factor (vWf). Here, we resolved the complete cDNA sequences of agglucetin subunits (alpha(1), alpha(2), beta(1) and beta(2)) by molecular cloning. Each cloned cDNA encoding the leader peptide (23 residues) and the mature subunit (131/135/123/126 residues) shares a high degree of homology to each other and the C-type lectin-like GP Ib-binding proteins (BPs). Furthermore, agglucetin specifically caused platelet agglutination and surface exposure of integrin alpha(IIb)beta(3) with a GPIb-dependent manner in washed platelets, based on the observation that the enhanced expression of functional alpha(IIb)beta(3) was suppressed by a GPIb-cleaving metalloproteinase, crotalin. Pretreating platelets with staurosporine or BAPTA-AM also completely blocked the exposure of functional alpha(IIb)beta(3), suggesting that the activation of protein kinase C and intracellular calcium mobilization are involved in the GPIb-dependent signaling. In human platelet-rich plasma (PRP), agglucetin elicited sequential biphasic responses of platelet agglutination and aggregation in a GPIb- and alpha(IIb)beta(3)-dependent manner, respectively, implying that other cofactors may amplify platelet activation to trigger aggregation.

Inhibition of neuropathic pain by a potent disintegrin—triflavin

Injury to peripheral nerves may result in severe and intractable neuropathic pain. Many efforts have been focused on the elucidation of the mechanisms of neuropathic pain. It was found here that integrin plays an important role in the induction of neuropathic pain and treatment of disintegrin is able to attenuate neuropathic pain. The rats were induced hyperalgesia by tightly ligating the L5 spinal nerve and cut just distal to the ligature on one side. Mechanical and thermal stimuli were applied in the middle dermatome of the hind paw. Epidural administration of triflavin (TFV), an arginine-glycine-aspartic acid (RGD) containing disintegrin, inhibited hyperalgesia induced by either mechanical or thermal stimulation. Immunohistochemistry showed that the sprouting of sympathetic nerves into DRG by neuropathic surgery was markedly inhibited by TFV. Beta 1 integrin mRNA of L5 DRG increased immediately 1 day after tight ligation and cut of L5 spinal nerve. However, beta 1 integrin mRNA in uninjured L4 DRG increased later on Day 3 after surgery. On the other hand, alpha-CGRP precursor mRNA decreased in ipsilateral L5 DRG but increased in L4 DRG after neuropathic surgery. Immunohistochemistry shows that beta 3 integrins of L5 as well as L4 increased in response to neuropathic surgery and administration of triflavin antagonized the increasing action. These results suggest that there is interaction between injured and uninjured neurons and the induction of neuropathic pain is related to neuronal sprouting. Disintegrin is able to inhibit neuronal sprouting and the induction of hyperalgesia induced by peripheral nerve injury and may thus be a new category of drugs to be developed for the treatment of neuropathic pain.

CSF Protein Profiling by Surface-Enhanced Laser Desorption/Ionization Time-of Flight Mass Spectrometry after Depletion of Albumin and IgGs

Proteins in cerebrospinal fluid (CSF) could be used as indicators of some central nerve system diseases such as Alzheimers disease. However, the high abundant proteins in CSF make the analysis very difficult because those proteins tend to shade those of lower abundance. Usually, some of the low abundant proteins are biomarkers. Therefore, it could be a better way to deprive high abundant albumin and IgGs from CSF first and analyze the remaining in order to find some imperceptible changes of low abundant proteins. We applied anti-human albumin antibody combine Protein A/G-Sepharose beads to specifically deprive albumin from the CSF sample. With high affinity to IgGs, Protein A/G could also deprive IgGs in the same approach. We perform single immunoprecipitation experiment and deprive albumin and IgGs from the CSF. SELDI-TOF-MS, combine high through-put separation and analysis of proteins, could profile the sub-proteome of a subject within hours. This system is extremely sensitive and rapid method to analyze complex mixtures of proteins and peptides. The objective of this study was to utilize SELDI-TOF-MS to determine the specific protein markers of CSF. We setup the standard process procedure to reach the reliability and reproducibility of instrument output. Our results provide a rational basis for identifying biomarkers from CSF using SELDITOF-MS.

Identification of Genes Expression in Spinal Cord after Peripheral Inflammation Using DNA Microarray Techniques

Peripheral inflammation may result in the transcriptional activation of many genes in spinal cord dorsal horn neurons. However, reports of many studies suggested that more than genes should be involved in spinal nociceptive processing after peripheral inflammatory stimulation. In this study, we used microarray techniques to identify genes that were altered in spinal cord after peripheral inflammation. Male rats received an injection of complete Freunds adiuvant (CFA) (1:1, CFA: saline, 200 μ1) into the plantar surface of the left hindpaw. 24hrs after injection, L4-L5 of spinal cord segments were removed and tissues were cut along the midline into ipsilateral and contralateral sides. Total RNA was isolated and synthesis of eDNA and cRNA were carried out. 10 μ g of ipsilateral side (n=4) or contralateral side (n=2) cRNA were hybrid with each chamber of a CodeLink Rat whole genome microarray chip, which contains approximately 33,000 oligonucleotide gene probes. The changes in gene identified through the microarrays analysis were confirmed with quantitative RT-PCR. We screened out total 110 genes that were altered over 2-fold in ipsilateral side comparing contralateral side. Among them, 62 genes including 32 of EST/unknown genes were shown increase and 48 genes including 16 of EST/unknown genes were shown decrease in level of expression. Our results suggest that peripheral inflammation induced the dynamic activation of neuron markers in nociceptive processing of spinal neurons.