Table of Content

25 September 2004, Volume 24 Issue 3

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Effect of Tetramethylpyrazine on P2X₃ receptor Mediated Excitation of Nociceptive Afferents

LIANG Shang-dong, GAO Yun,XU Bao-hua, MU Song-niu, XU Chang-shui

2004, 24(3):  131-133. 

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Using a behavioral study to characterize the effects of TMP administered by intraplantarlly and intrathecally on the acute nociception（flinching） mediated by P2X receptor activation of rat hindpaw. The P2X₃ receptor antagonist TNP-ATP（0.3μmol/L） significantly depressed the acute nociception mediated by ATP （1 μ mol/L） and α,β -meATP （0.6μmol/L） in rat hindpaw. Intraplantar administration of TMP （0.1-10 mmol/L） inhibited the acute nociception in a dose-dependent manner mediated by ATP （1 μ mol/L） or α，β -meATP （0.6μmol/L） in rat hindpaw. Intrathecal ly applied TMP （50 mmol/L） inhibited the nociception induced by ATP （1 μmol/L） or α , β -meATP （0.6 μ mol/L） in rat hindpaw. TMP inhibited the nociceptive behaviors induced by P2X₃ receptor mediated excitation of nociceptive afferents.

Modification on Enucleation Method of Porcine Oocytes

LIU Dong1,2, TANG Lin-lin1,2, CHEN Xiao-yu3, ZHANG De-fu1,2

2004, 24(3):  137-137. 

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The study on porcine somatic nuclear transfer is of the great meaning in the field of biomedicine. This experiment was conducted to study enucleated method, which is one of the key techniques for animal cloning. The results show that the modified Willadsen’s method for enucleation, called two-step-squeezing method, is significantly better than McGrath-Solter’s method in the enucleated rates （73.9%, 42.5%,P＜0.05） and the rates （81.2%, 67.4%, P＜0.05） of absolutely removing nuclei matter. It was also found that the rates （89.1%） of absolutely removing nuclei matter for 36h-matured-in-vintro oocytes is significantly higher than that for 44h group （55.8%, P＜0.05）.

Expressions of Bcl-2 and Bcl-xl Proteins after Cerebral Ischmia/Reperfusion Injury Induced by Precondition in Rat Hippocampus

XU Fu-yu,YIN Zhi-sheng, LIU Wen-jun, CUI Min, CHEN Lian-bi,ZHAO Hong-ling

2004, 24(3):  138-141. 

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To study the function of apoptosis-blocking proteins Bcl-2 and Bcl-xl in the neuroprotective role of ischemic preconditioning （IPC）, cerebral ischemia/Reperfusion rat model was established by using 4-vessel occlusion and repassing, the histopathological changes, the percentage of apoptosis and the expression of Bcl-2 and Bcl-xl proteins of the neurons in CA1 region of rat hippocampus with IPC were examined and detected with Nissls staining, Flow cytometry （FCM） and immunohistochemistry technique. The results showed that forebrain ischemia and reperfusion can cause apoptosis of neurons in CA1 region of hippocampus. IPC protects the neurons against the lethal ischemia/reperfusion injury by decreasing the number of apoptotic neurons, and the expressions of Bcl-2 and Bcl-xl are induced by IPC in early period of reperfusin. The results suggest that blocking the process of neuronal apoptosis after ischemic reperfusion may play an important role in the mechanism of neuroprotection induced by IPC.

Study On the Method of Acutely Isolating Hippocampal Neuron in Rat and Patch Clamp

WANG Xi-jie,CHENG Hong, GAO Rong,CHENG Jie, XIAO Hang

2004, 24(3):  142-145. 

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The hippocampal neurons were isolated by enzymatic and mechanical methods from 10-15 day postnatal rat, and its activity were verified by morphology and electrophysiology. Results showed that single hippocampal neuron with good shape and physiological characteristic can be obtained by using this method. After blocked Na⁺channels by 0.5μmol/L TTX, standard whole cell voltage dependent potassium current and delayed rectifier potassium current were recorded. This proved that the method is suitable for study on hippocampal neurons by patch clamp,and it have important value for deeply research the effects of drugs and toxins on hippocampal ion channels and on its signal transduc-tion mechanism.

Establishment of Renal Transplantation Model in Beagle Dog

YING Hua-zhong1,PAN Yong-ining1,ZOU Yu2,XU Xiao-ping1,HUANG Hong-feng2,LI Xiong-feng3, WANG Hui1，XU Jian-qin1, WANG De-jun1, ZHOU Wei-min1, CHEN Min-li1

2004, 24(3):  146-148. 

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A simple and reliable renal transplantation model was established in Beagle dogs performed with allograft, and postoperative serum creatinine, blood stream and general examination of transplanted kidney were observed.The results showed that the serum creatinine and BUN of the model dogs were risen rapidly 3 days after operation. The serum creatinine was varied from 61.1±13.5 μ mol/L to 1153.9 ± 124.4μ mol/L, and BUN was varied from 3.68±0.62 mmol/L to 24.23 ± 7.17 mmol/L, but the serum creatinine and BUN of the shamed group were remained steadily. So typical changes of acute renal rejection in Beagle dog renal transplantation could be observed in allograft by serum creatinine, blood stream and general examination of transplanted kidney. It suggested that Beagle dog renal transplantation model was reliable, simple and stable, it can be used as a model to study transplantation immunity and evaluate immunosuppressant.

Establishment and Evaluation of Collagen -induced Arthritis Model in Mice

YAN Shang-xue ,CHEN Feng-qin, HUANG De-wu, GUO Yan-song

2004, 24(3):  149-152. 

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To establish the model of collagen-induced arthritis （CIA） and method of evaluation, the mice were injected subcutaneously with CFA （complete Freund’s adjuvant） and the change of secondary paw-swelling were observed and scored. The level of PGE was assayed by spectrophotometer； and the ³H-TdR incorporation was used to detect ConA- and LPS- induced splenocytes proliferation and the production of interleukin （IL）-1 and IL-2. Hepatic pathological changes was examined by Hematoxylin-eosin （HE） stain method. The results showed that the inflammation and paw swelling of treated mice increased significantly compared with those of control. Meanwhile, the ConA-induced splenocytes proliferation,PGE, IL-1 and IL-2 production were increased.The pathological examination showed that synovial cell increased ,pannus formed and inflammatory cells infiltrated into synovium. The results indicated that CIA mice model can be established by subcutaneous injection of CFA.

Investigation on Final Host and Geographic Distribution of Paragonimus in Fujian Province

LI You-Song, ZHANG Shi-Yang, XU Long-Shan, ZHOU Xian-Min, LIN Jin-Xiang

2004, 24(3):  153-156. 

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To investigate and summarize the species and distribution in their counties of the final host of Paragonimus in Fujian Province, from captive wild animals,or collected animal soil and encysted metacercarias of Paragonimus were separated to identify species firstly and infect cats,dogs and rats.When eggs were examined,they were anatomized to check adult worms for making tinct samples and judging species. The final host animals of 7 species of Paragonimus found in the whole province were 18,with the most of civet cat family, cat family,dog family and rat family.

Acute Toxicity and Dermal Sensitzation of Bacillus thuringiensis Original Drug

LIU Zhi-Yong, LI Qi-Fu, ZHOU Yin-Ping, DAI Li-Guang, ZHOU Ming, LIU Lin, TIAN Ze-Min, BAO Jin-Ying, YU Rong-Zhen, YANG Xin-Yue

2004, 24(3):  157-159. 

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According to toxicological test methods of pesticides for registration（GB 15670-1995）,The test of acute toxicity of biopesticide - bacillus thuringiensis, include acute oral toxicity, acute dermal toxicity, acute inhalation toxicity, acute eye irritation test and dermal sensitization were carried out. The acute oral toxicity LD₅₀ of this biopesticide was 3830 mg/kg in male and 3160 mg/kg in female, acute dermal toxicity LD₅₀（4h）>2 150 mg/kg, LC₅₀（2h）>5 000 mg/m³, there is no irritation to rabbit eye, dermal sensitization：rate of sensitization is zero.So the acute oral toxicity, acute dermal toxicity and acute inhalation toxicity of bacillus thuringiensis are lower. There is no irritation to rabbit eye, and it belongs to grade I weak sensitizing matter.

Experimental Investigation of P2 Receptors and Pain

XU Chang-Shui, LIANG Shang-Dong

2004, 24(3):  167-171. 

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Receptors for purines and pyrimidines （P receptors ） could be subdivided into P1 （adenosine） receptors and P2 receptors. P2 receptors were activated preferentially by ATP、UTP and analogs, and P2 receptors subtypes were placed in two major families： P2X （ligand-gated ion channels, LGICs） and P2Y （G protein-coupled receptors, GPCRs）.Endogenous nucleotide mediate a lot of functions via cell P2 receptors In many organs, and P2 receptors have crucial mediated roles in the pathologic status of pain （such as neuropathic pain、actuepain、inflanmm-atory pain、visceral pain, and so on）.The P2X₃-like immunoreactivity and functional expression in sensory neurons and using P2X₃ receptor gene knockout and antisense oligonucleotide technologies, A-317491（a novel, selective P2X₃ antagonist）, demonstrate a crucial role of ATP and P2X₃ receptors in pain； the heteromeric P2X_2/3 receptor is also involved in pain. In addition, several subtypes of P2Y receptor are also expressed in sensory neurons. P2Y receptors might also be relative with pain.P2Y₁ and P2Y₂ receptors could generate excitability as nociception, while P2Y₆ being antinociceptive. Agonists and antagonists for P2 receptors may provide novel drugs in patho-logic status such as pain.