Objective: To observe the changes of pain thresholds and P2X3 receptor expression in spinal cord of chronic constriction injury of the sciatic nerve model rats by unilateral and bilateral acupoints electroacupuncture treatment. Methods: The difference value of pain threshold was compared before and after the electroacupuncture by determining the 15th and fifth day pain threshold of rats after the model was successfully made. And the expressing of P2X3 receptor in spinal dorsal horn was detected by using immunohistochemistry. Results: ①In electroacupuncture groups,difference values of the pain threshold of rats were significantly increased compared with those of the model control group( P ﹤ 0. 01). In bilateral acupoints electroacupuncture group,rats' difference values of pain threshold all had significant differences compared separately with that of contralateral and ipsilateral acupoints electroacupuncture groups( P ﹤ 0. 01). Rats' difference values of pain threshold between contralateral and ipsilateral acupoints electroacupuncture groups had no significant difference( P 0. 05). ②In electroacupuncture groups,P2X3 receptor positive material expression in spinal dorsal horn of rats was decreased in different levels compared with that of model control group( P ﹤ 0. 01). Compared among electroacupuncture groups,P2X3 receptor expression in spinal dorsal horn of rats had no significant difference( P 0. 05). Conclusion: Electroacupuncture therapy may produce analgesia by suppressing P2X3 receptor expression in the spinal dorsal horn of CCI rats. When using electroacupuncture to treat chronic constriction injury pain of the sciatic nerve,bilateral acupoints have a stronger analgesic effect than unilateral acupoints.

Objective:To compare the expression changes of Substance P gene in spinal cord of chronic constriction injury of the sciatic nerve model rats by sick side,healthy side and bilateral sides electro-acupuncture treatment.Methods:After CCI model was successfully made,35 male Sprague-Dawley rats were randomized into false group(Group J),control group(Group A),sick side electro-acupuncture group(Group B,),healthy side electro-acupuncture group(Group C),bilateral sides electro-acupuncture group(Group D).Each electro-acupuncture group was treated for 9 days,then the changes of pain threshold before and after the electro-acupuncture were observed;and the expression of SP gene in spinal dorsal horn was detected by using reverse transcriptase-polymerase chain reaction.Result:Compared among Group B,C and D:the value of Z/R Reinheit Zahl of SP gene and β-actin was:Group D<Group B<Group C,there was difference between each other(P<0.05).Analyzing correlation between SP gene and difference value of the pain threshold:sp gene expression in rats spinal dorsal horn and difference value of the pain threshold had obvious statistic correlationship(P<0.01).Conclusion:Electro-acupuncture applied separately in ST36 and GB 34 on sick side,healthy side and bilateral sides,generated the analgesia function by suppressing SP gene expression in rats spinal dorsal horn.

Objective: To study the variation of expression of CD4+ CD25+ regulatory T cells and Foxp3 mRNA in patients infected with Helicobacter pylori and its clinical significance.Methods: Changes of expression in 35 H.pylori-negative adults and 45 H.pylori-positive patients and Foxp3 mRNA were analyzed using immunohistochemistry and PCR.Results: The proportion of CD4+ CD25+ regulatory T cells in the gastric mucosa in H.pylori-positive patients(5.012%)was obviously much higher than that of CD4+ CD25+ regulatory T cells in H.pylori-negative patients(0.739%).The level of expression of Foxp3 mRNA was significantly higher than those of H.pylori-negative patients(P <0.01)in gastric mucosa of patients infected with Helicobacter pylori.Conclusion: The patient infected by H.pylori may inhibit specific T-cell response caused by H.pylori through the expression of the up-regulation of CD4+ CD25+ regulatory T cells,resulting in the long-term colonization of H.pylori.

目的 观察幽门螺杆菌(helicobacter pylori, Hp)感染及根除后CD4+CD25+调节性T细胞的表达变化,以探讨Hp长期定植在胃黏膜引起慢性持续性感染的原因.方法 采用免疫组化法分析45例Hp阳性患者(研究组)Hp清除前后及35例Hp阴性患者(对照组)胃黏膜中的CD4+CD25+调节性T细胞的表达变化. 结果 Hp阳性患者局部胃黏膜中的CD4+CD25+调节性T细胞明显增多,占5.012%,明显高于Hp阴性患者胃黏膜中的CD4+CD25+调节性T细胞,占0.739%,差异有统计学意义 (P＜0.01).根除Hp治疗后CD4+CD25+调节性T细胞明显减少,占0.551%,明显低于清除前,差异有统计学意义(P＜0.01). 结论 Hp感染后可能通过上调CD4+CD25+调节性T细胞的表达,抑制Hp引起的特异性T细胞反应,造成Hp的长期定植.根除Hp后人胃黏膜内CD4+CD25+调节性T细胞明显减少可能是Hp被清除后,该类细胞凋亡或者移出胃黏膜所致.

BACKGROUND/AIMS:H. pylori persists for the virtual life of its host. Recent studies suggested that CD4+ CD25+ regulatory T cells may be involved in this process. However, the alteration of CD4+ CD25+ regulatory T cells after eradication of H. pylori remains a question.METHODOLOGY:By using biopsies from 45 H. pylori-positive patients and the ones after eradication of H. pylori and 35 H. pylori-negative adults, real-time PCR and general PCR were used to quantify the expression of Foxp3 mRNA. IHC was used to semi-quantify the number of CD4+ CD25+ T cells in gastric mucosa.RESULTS:We found that proportion of CD25+ T cell in CD4+ T cells accounted for 0.739% in H. pylori-negative individuals, while it was accounted for 5.012% in H. pylori-positive patients. After eradication of H. pylori, proportion of CD25+ T cell in CD4+ T cells declined (p < 0.01) and it accounted for 0.551%. The level of Foxp3 mRNA significantly decreased (p < 0.01) in gastric mucosa of patients after eradication of H. pylori.CONCLUSIONS:CD4+ CD25+ regulatory T cells decreased in gastric mucosa when patients received eradication of H. pylori. Eradication of H. pylori results in the significant descrease of Foxp3 mRNA in gastric mucosal, or using the drugs of anti-H. pylori induce the reduction of gastric mucosal Foxp3 mRNA expression, which is the a key regulatory gene for the development and function of CD4+ CD25+ regulatory T cells, thus contributing to the eradication of H. pylori. All the data offer new possibilities that Foxp3 gene may be the new target of immunization intervention strategies for eradication of H. pylori.

Objective To investigate the effect and the potential mechanism of splenic artery coarctation on the expression of iNOS and Th1/Th2 cytokines in spleen of cirrhotic rats with portal hypertension (PHT). Methods Cirrhotic rats were randomized into 3 groups (n= 10):sham operation group (SOG), splenic artery coarctation group (SAC) and splenic artery ligation group (SAL). Ten normal rats treated with sham operation were employed to serve as normal control group (NCG). Immunohistochemial staining was used to observe iNOS. RT-PCR was used to detect IFN-γ and IL-4mRNA. The Pearson's correlation analysis was used to investigate the relationship between iNOS and IFN-γ or IL-4. Results The expression of iNOS was increased significantly in spleen of cirrhotic rats as compared with NCG(P＜0. 01). It was decreased after SAC and SAL compared with SOG (P＜0. 01). The expression of IFN-γmRNA and IFN-γ/IL-4 of SOG were decreased but IL-4mRNA increased significantly than that of NCG(P＜0.01). IFN-γmRNA was increased after SAC compared with SOG (P＜0.05). IL-4mRNA was decreased and IFN-γ/IL-4 increased after SAC and SAL compared with SOG (P＜0. 05). The expression of iNOS was negatively correlated with the expression of IFN-γmRNA(r=-0.672, P＜ 0.01 ) and positively correlated with the expression of IL-4 mRNA (r=0.634,P＜0. 01). Conclusion The expression of iNOS is decreased and IFN-γ/IL-4 increased after SAC in spleen of cirrhotic rats with PHT and it may improve Th1/Th2 polarization by reducing the expression of iNOS.

目的 观察脾动脉血流减少对肝硬化门静脉高压大鼠脾脏Toll样受体4(Toll-like receptor 4,TLR4)表达和巨噬细胞(Macrophage,Mφ)吞噬活性的影响.方法 肝硬化门静脉高压大鼠模型诱导成功后随机分为肝硬化组(MCG组)、脾动脉缩窄组(SAC组)和脾动脉结扎组(SAL组),每组10只,另10只正常大鼠为对照组(NCG组).术后4周处死大鼠,用免疫组化法测定脾脏TLR4表达量,鸡红细胞吞噬实验检测脾脏Mφ吞噬活性,就脾脏TLR4表达量和Mφ吞噬活性进行相关分析.结果 脾脏TLR4表达量:MCG组明显高于NCG组[(210.83±42.57)vs(63.08 ±18.53)],差异有统计学意义(P<0.01);SAC组(180.03±29.67)、SAL组(170.83±32.65),均明显低于MCG组.差异有统计学意义(均P<0.05).脾脏Mφ吞噬率和吞噬指数:MCG组明显高于NCG组[(35.83%±4.95%)vs(17.32%±3.51%)]、[(0.63±0.12)vs(0.22±0.08)],差异有统计学意义(均P<0.05);SAC组(23.46%±3.31%)、(0.29±0.05)和SAL组(22.34%±2.95%)、(0.23±0.07),均明显低于MCG组,差异有统计学意义(均P<0.05).脾脏Mφ吞噬率、吞噬指数与TLR4表达量呈显著正相关(均P<0.01).结论 肝硬化门静脉高压大鼠脾脏TLR4表达量增高,Mφ吞噬活性增强,脾动脉缩窄和结扎能减弱脾脏Mφ吞噬活性,这可能与术后脾脏TLR4表达量减少有关.

Objective:To observe the expression changes of Substance P in spinal cord of chronic constriction injury of the sciatic nerve model rats by single side and bilateral side electro-acupuncture.Methods:After CCI model was successfully made,seventy male Sprague-Dawley rats were randomized into false group(Group J),control group(Group A),sick side electro-acupuncture group(Group B),healthy side electro-acupuncture group(Group C),bilateral side electro-acupuncture group(Group D).After every electro-acupuncture group was treated 9 days,observe the changes of pain threshold before and after the electro-acupuncture;And detected the expressing of SP in Spinal dorsal horn by using immunohistochemistry.Results:Compared within group B.C.D,The total IOD value of SP positive material in Spinal dorsal horn of rats in Group D decreased to various degrees compared with Group B and C(P<0.01);The total IOD value of SP positive material in Spinal dorsal horn in rats of Group B was less than Group C(P<0.01).Conclusion:Electro-acupuncture therapy may be through suppressing substance P content in rats spinal dorsal horn;and generated the analgesia function,within all groups,bilateral side electro-acupuncture has the most obvious analgesic effect.

As a crucial bioactive substance,Nitric Oxide(NO)is considered to play a key role in the pathogenesis of portal hypertension of liver cirrhosis.Nitricoxide synthase(NOS)is a key enzyme for NO.Recent researches show that interventional measures on NOS to regulate the local NO concentration,such as the liver and splanchnic vessels,have achieved curative effects.This review was concerned on the differential expression of NOS and its significance among hyperdynamic circulation,cirrhotic liver,splenomegaly and immunologic derangement,which may shed light on the mechanisms of liver cirrhosis with portal hypertension and be of help to develop new therapies.

肝硬化动物模型的制作是肝硬化疾病研究的重要基础,目前制作方法较多,其中以CCl4诱导法最为普遍,但由于大鼠对CCl4敏感性的个体差异导致模型制作时死亡概率高、成型率低.本研究通过监测大鼠体重变化,了解大鼠对四氯化碳反应的个体差异,并根据体重变化调整CCl4剂量,以减少大鼠死亡概率,提高模型成功率。

Objective:By investigating the key point of sensory plane examination and the corresponding acupoints of spinal cord, to evaluate the applicability range of the segmental innervation law, one of the basic laws in clinical acupuncture, in the examination and treatment of spinal cord injury. Methods:Observing the relationship between the key point of sensory plane of spinal cord and the corresponding acupoints.Clarifying the muscle distribution under the related acupoints. And identifing the segmental innervation according to the corresponding acupoints. Results:The acupoints of the trunk consistent with the segmental innervation law, but the acupoints of the distal end of four limbs, which have specific therapeutical effect, have other laws of acupunture. Conclusion: Key points of dermatome and acupoints have several coincidences and their relationship deserves further research.

Objective:To investigate the nerve segment dominate regularity of muscular layer in acupoints of twelve meridian in the distal limbs, study the application scope of Spinal Segment Dominated Principleof clinic acupuncture principle in further. Methods: Observe the regularity of muscles of each layer dominated by corresponding myelonic nerve segment, according to the anatomical distribution of the muscles of acupoints of twelve meridian in distal limbs. Conclusion: Comparing to the body acupoints, points in the distal limbs, which have relatively specific therapeutic effect, have nothing to do with the Spinal Segment Dominated Principle, may have other regularity.