The cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signaling acts a pivotal part in hyperpolarization-activated cyclic nucleotide-gated (HCN) channels-mediated neuropathic and inflammatory pain. However, there has been no evidence of cAMP-PKA signaling is involved in regulation of spinal HCN channels function in the occurrence of diabetic neuropathic pain (DNP). The study aimed to elucidate the impact of HCN channels on neuropathic pain in a rat model of diabetes induced by streptozotocin, and whether cAMP-PKA signaling is involved in regulation of HCN channels function. In this report, we evaluated the effect of intrathecal administration of HCN channel blockers ZD7288, cAMP inhibitor SQ22536 and PKA inhibitor H-89 on nociceptive behavior in DNP rats. The mechanical withdrawal threshold (MWT) was measured to evaluate pain behavior in rats. Protein expression levels of HCN2, HCN4 channels and PKA in the spinal dorsal horn of rats were assessed. Furthermore, the levels of cAMP in rat spinal dorsal horn was analyzed. We discovered that DNP rats showed significant mechanical allodynia and are related to the increased HCN2 and HCN4 channels expression, enhanced cAMP production and elevated the expression of PKA protein in the spinal dorsal horn, which were attenuated by intrathecal ZD7288. Furthermore, intrathecal injection of SQ22536 and H-89 significantly reduced the HCN2 and HCN4 channels expression in the spinal dorsal horn of DNP rats. Our findings indicate that HCN channels of the spinal dorsal horn participate in the pathogenesis of allodynia in rats with DNP, which could be regulated by cAMP-PKA signaling. Therefore, HCN channels and cAMP-PKA signaling are potential targets for hyperalgesia treatment in DNP patients.

大黄为临床常用中药,药用历史悠久,功效独特.在历代中药本草典籍中均有记载,始载于《神农本草经》.其来源为蓼科植物掌叶大黄Rheum palmatum L .?唐古特大黄Rheum tanguticum Maxim .ex Balf .或药用大黄Rheum officinale Baill .的干燥根及根茎[1].

目的 观察鞘内注射超极化激活环核苷酸门控通道(HCN通道)拮抗剂ZD7288对STZ诱导的I型糖尿病大鼠的热痛阈及血糖的影响,探讨背根节HCN通道在糖尿病周围神经痛(DPN)中的作用.方法 4～5周龄雄性SD大鼠随机分为3组:对照组、STZ组(鞘内给予生理盐水)和ZD7288+STZ组(鞘内给予ZD7288).单次腹腔注射链脲佐菌素(STZ,60 mg/kg)制备I型糖尿病大鼠模型,分别于注射STZ前1d及注射后7、14、21、28d测定空腹血糖、热缩足反射潜伏期(TWL),免疫印迹技术检测背根节HCN1,2通道的表达.结果 与对照组相比,大鼠腹腔注射STZ 2周后空腹血糖显著升高,TWL降低,背根节HCN1,2通道表达显著增高(P＜0.01);鞘内给予ZD7288可显著延长糖尿病大鼠TWL,降低背根节HCN1,2表达(P＜0.01),但对空腹血糖无显著影响.结论 背根节HCN通道表达增加可能促进了糖尿病周围神经痛的发生,鞘内给予HCN通道拮抗剂具有良好的镇痛效应,但对血糖无明显改善作用.

目的:观察脊髓背角超极化激活环核苷酸门控通道4(HCN4)在糖尿病神经痛(DNP)发生与发展过程中的作用.方法:选择4～5周龄SD大鼠,随机分为3组:对照组、链脲佐菌素STZ组和ZD7288+ STZ组.单次腹腔注射60 mg/kg STZ制备大鼠糖尿病模型,注射STZ 2周后鞘内给予HCN通道拮抗剂ZD7288.采用yonFrey丝测定机械性缩足反射阈值(MWT),Western Blot法检测大鼠脊髓背角HCN4通道表达,ELISA法检测背角cAMP含量.结果:大鼠腹腔注射STZ 2周后MWT明显缩短,产生糖尿病神经痛,鞘内给予HCN通道拮抗剂ZD7288可显著延长糖尿病大鼠的MWT;DNP大鼠脊髓背角HCN4通道表达及cAMP含量显著增高,鞘内注射ZD7288可显著降低DNP大鼠脊髓背角HCN4通道表达及cAMP含量的升高.结论:脊髓背角HCN4通道表达增加促进了糖尿病神经痛的发生和发展,cAMP含量增加可能参与了HCN4通道的作用.

Objective:To investigate the role of cannabinoid receptor 1 (CB1 R) in neuropathic pain induced by sciatic chronic constriction injury (CCI) and its effect on the expression of hyperpolarization-activated cyclic nucleotide-gated (HCN) channels 2 in dorsal root ganglion.Methods:SD rats (7 ～ 8 weeks old) were divided into 4 groups.(1) sham group;(2) CCI group;(3) CP55940 + CCI group;(4) AM251 + CP55940 + CCI group.Mechanical withdrawal threshold (MWT) was measured.The expression of HCN2 channels was detected by Western Blot.Results:MWT of CCI rats was significantly lower than that in the sham group (P ＜ 0.05).Intrathecal administration of CP55940 (0.05 mg/kg),cannabinoid receptors agonist,markedly inhibited the decrease of MWT after nerve injury (P ＜ 0.05).Intrathecal administration of AM251 (0.05 mg/kg),CB1 R antagonist,markedly blocked the analgesic effect of CP55940.The expression of CB1 R and HCN2 channels in the injured L4,L5 and L6 dorsal root ganglion markedly enhanced on 7 d (P ＜ 0.05) and 14 d (P ＜ 0.05) after CCI.Intrathecal administration of CP55940 dramaticlly inhibited the increase of HCN2 channels expression after CCI.The inhibitory effect of CP55940 was blocked by AM251 (P ＜ 0.05).Conclusion:Activation of CB1 R has analgesic effect on neuropathic pain induced by CCI.The analgesic effect of CB1 R may be related to its inhibition of HCN2 channels expression in dorsal root ganglion.

阳离子氯离子联合转运蛋白(cation-Cl-cotransporter,CCC)是一组转运Na+、K+、Cl-进出细胞的膜蛋白.其中,钾氯联合转运蛋白2(K+-Cl-cotransporter 2,KCC2)和钠钾氯联合转运蛋白1(Na+-K+-C1-cotransporter 1,NKCC1)是调节神经系统氯离子稳态的主要CCC,NKCC1将CI-转运入胞内,KCC2将C1-转运至胞外,两者共同调节氯离子平衡及神经元的兴奋性[1].近年来越来越多的研究表明,KCC2/NKCC1在伤害性信息的传递过程中发挥重要的调节作用,其表达及功能异常促进了病理性疼痛的发生与发展,本文就KCC2/NKCC1与病理性疼痛关系的最新研究进展作一综述,以期深入理解KCC2/NKCC1在病理性疼痛发生与发展中的作用.

Objective:To investigate the role of cannabinoid receptorl (CB1 R) in sciatic chronic constriction injury (CCI) induced neuropathic pain and its effect on hyperpolarization-activated cyclic nucleotide-gated (HCN) channels 4 expression in spinal dorsal horn.Methods:7 ～ 8 weeks old SD rats were divided into 4 groups:(1) sham group;(2) CCI group;(3) CP55940 + CCI group;(4) AM251 + CP55940 + CCI group.Mechanical withdrawal threshold (MWT) were measured by electric von Frey pain detector.The expression of HCN4 channels was detected by Western Blot.Results:MWT of CCI rats was significantly lower than that in the sham group on 1 ～ 14 d after nerve injury (P ＜ 0.05).Intrathecal administration of CP55940 (0.05 mg/kg),cannabinoid receptors agonist,markedly inhibited the decrease of MWT after CCI (P ＜ 0.05).Intrathecal administration of AM251 (0.05 mg/kg),CB1 R antagonist,markedly blocked the analgetic effect of CP55940.The expression of HCN4 channels in the injured L4,L5 and L6 dorsal spinal cord was markedly enhanced after CCI (P ＜ 0.05).Intrathecal administration of CP55940 dramaticlly inhibited the increase of HCN4 channels expression after CCI.The inhibitory effect of CP55940 was blocked by CB1 R antagonist AM251 (P ＜ 0.05).Conclusion:Activation of CB1 R has analgesic effect to neuropathic pain induced by peripheral nerve injury.CB1 R's analgesia may be related to its inhibition of HCN4 channels expression in dorsal spinal cord.

超极化激活环核苷酸门控的阳离子通道(HCN通道)属于孔环阳离子通道家族成员.哺乳动物HCN通道存在4个亚型(HCN1-4),主要表达于心脏和神经系统.HCN通道激活后产生的超极化激活电流具有影响细胞膜静息电位、控制细胞兴奋性、调节心脏和神经节律等诸多生理功能,其表达及功能的异常改变与癫痫、脑缺血、病理性疼痛及心律失常等多种疾病的发生密切相关.HCN通道在糖尿病周围神经病变(包括糖尿病感觉性周围神经病变及心脏自主神经功能障碍)发生中扮演着重要角色.

Spinal cannabinoid receptor 1 (CB1R) and purinergic P2X receptors (P2XR) play a critical role in the process of pathological pain. Both CB1R and P2XR are expressed in spinal dorsal horn (DH) neurons. It is not clear whether CB1 receptor activation modulates the function of P2X receptor channels within dorsal horn. For this reason, we observed the effect of CP55940 (cannabinoid receptor agonist) on ATP-induced Ca2+ mobilization in cultured rat DH neurons. The changes of intracellular calcium concentration ([Ca2+]i) were detected with confocal laser scanning microscopy using fluo-4/AM as a calcium fluorescent indicator. 100 μM ATP caused [Ca2+]i increase in cultured DH neurons. ATP-evoked [Ca2+]i increase in DH neurons was blocked by chelating extracellular Ca2+ and P2 purinoceptor antagonist PPADS. At the same time, ATP-γ-S (a non-hydrolyzable ATP analogue) mimicked the ATP action, while P2Y receptor agonist ADP failed to evoke [Ca2+]i increase in cultured DH neurons. These data suggest that ATP-induced [Ca2+]i elevation in cultured DH neurons is mediated by P2X receptor. Subsequently, we noticed that, in cultured rat DH neurons, ATP-induced Ca2+ mobilization was inhibited after pretreated with CP55940 with a concentration-dependent manner, which implies that the opening of P2X receptor channels are down-regulated by activation of cannabinoid receptor. The inhibitory effect of CP55940 on ATP-induced Ca2+ response was mimicked by ACEA (CB1R agonist), but was not influenced by AM1241 (CB2R agonist). Moreover, the inhibitory effect of CP55940 on ATP-induced Ca2+ mobilization was blocked by AM251 (CB1 receptor antagonist), but was not influenced by AM630 (CB2 receptor antagonist). In addition, we also observed that forskolin (an activator of adenylate cyclase) and 8-Br-cAMP (a cell-permeable cAMP analog) reversed the inhibitory effect of CP55940, respectively. In a summary, our observations raise a possibility that CB1R rather than CB2R can downregulate the opening of P2X receptor channels in DH neurons. The reduction of cAMP/PKA signaling is a key element in the inhibitory effect of CB1R on P2X-channel-induced Ca2+ mobilization.

目的:观察小隐静脉及大隐静脉曲张泡沫硬化剂注射治疗的疗效.方法:回顾性分析88例小隐静脉及大隐静脉曲张泡沫硬化剂注射治疗患者的临床资料.结果:患者治疗满意,无明显复发.5例患者2次注射治疗,1例患者深静脉血栓形成,13例患者患肢肿胀,无肺栓塞.结论:对小隐静脉及大隐静脉曲张泡沫硬化剂注射治疗,具有方便、经济、恢复快、影响小、微创等优点.

目的:探讨彩色多普勒引导下乳腺肿块微创旋切术的应用价值.方法:收治乳腺肿块女性患者126例,采用彩色多普勒引导下乳腺肿块微创旋切术治疗,观察治疗效果.结果:126例患者手术均顺利完成.术后恢复好,术后6个月复查无肿物残留.结论:彩色多普勒引导下乳腺肿块微创旋切术的应用价值显著.

目的:观察大麻素对背根节神经元ATP诱发的[Ca2+]i升高的影响及机制.方法:培养SD大鼠背根节神经元,采用激光共聚焦技术检测培养神经元[Ca2+]i的变化.结果:ATP(100 μmol/L)经P2X受体介导可导致培养的背根节神经元[Ca2+]i增高(P＜0.05);大麻素受体激动剂CP55940预孵育10 min可剂量依赖性地抑制背根节神经元ATP所致的[Ca2+]i升高(P＜0.05);CB1受体(cannabinoid receptor 1,B1R)的拮抗剂AM251(10μmol/L)、CB2受体(Cannabinoid receptor 2,CB2R)的拮抗剂AM630(10 μmol/L)均可显著降低CP55940(1 μmol/L)的抑制效应(P＜0.05);腺苷酸环化酶激动剂Forskolin(10 μmol/L)可逆转CP55940对ATP的抑制作用(P＜0.05).结论:CP55940可显著抑制背根节神经元ATP诱发的[Ca2+]i升高,CP55940的抑制效应可能是由CB1、CB2受体介导抑制背根节神经元PKA活性所致.

To foster college students’ innovative capability has been a central topic of teaching reform . The experimental teaching reform ,however ,involves many aspects at teaching work ,such as heavy workload ,cumbersomeness and complexity .The project carried out the reform of functional experi-ment from several aspects ,including experimental projects ,experimental teaching means and meth-ods ,experimental teaching materials ,experimental teaching quality control and evaluation system , etc .The study will have positive significance to train high-quality medical application-oriented talents with practical ability and innovative spirit .

目的 分析腹腔镜阑尾切除术的方法、经验及注意事项.方法 总结我院2009年11月至2013年11月期间收治的458例阑尾炎患者的临床资料,慢性阑尾炎20例,急性单纯性阑尾炎108例,急性化脓性阑尾炎210例,急性坏疽性、穿孔性阑尾炎120例.结果 成功完成腹腔镜阑尾切除术452例,中转开腹阑尾切除术6例,无严重并发症发生.结论 腹腔镜阑尾切除术具有操作不复杂、术野清晰、探查范围广、切口小、美观、术后恢复快、术后并发症少,感染率低等优点,对诊断不明确的患者、异位阑尾患者、肥胖患者,腹腔镜阑尾切除术有不可替代的优势,适合在基层医院推广应用.

目的 探讨颈淋巴结结核的外科治疗.方法 106例均经手术及病理确诊的颈淋巴结结核患者,采用病灶清除+区域淋巴结清扫术106例(25例结节型,28例结节向脓肿转化型,51例寒性脓肿,2例窦道型).结果 采用病灶清除+区域淋巴结清扫术患者中104例治愈,切口均Ⅰ期愈合,未发生手术并发症,2例复发,住院5 ～21d.结论 颈淋巴结结核的外科治疗效果满意,可明显减少复发.

1病历摘要患者女,60岁,因右大腿肿痛1周于2010年12月3日入院.患者曾在当地医院行抗炎,局部外敷药物治疗,病情未见好转转入我院.否认既往有肝炎、伤寒、结核病史,但有精神病病史.体格检查:一般情况可,体形消瘦.双肺呼吸音清,未闻及干湿性啰音.腹胀,未见肠型、蠕动波.

Objective To investigate the effect of rhein on the hypertrophy of renal proximal tubular epithelial cells induced by high glucose in rats.Methods Studies were performed on anesthetized SD rats.Renal proximal tubular epithelial cells were gained by microdissection and were cultured in RPMI 1640 medium.The cell types were idenfied with immunocytochemistry.The cells were incubated with high glucose(30 mM) to induce cell hypertrophy.To observe the effcet of rhein on the hypertrophy induced by high guucose,the cells were cultured with different concentrations of rhein(30,15 and 5 mg/L) for 72 hr.Then the cell size,3H-lecuine incorporation,and cellular protein content were detected to observe the changes.Results High glucose(30 mM) resulted in enlargment of renal proximal tubular epithelial cells,increases of 3H-leucine incorpiration and cellular protein content significantly.On the contrary,rhein inhibited the hypertrophy of renal proximal tubular epithelial cells induced by high glucose.Rhein(30 mg/L) decreased cell size,3H-leucine incorporation and cellular protein content significantly.Rhein(15 mg/L) decreased 3H-leucine incorporation and cellular protein content.Rhein(5 mg/L) decreased cellular protein content.Conclusion Rhein can inhibit the hypertrophy of renal proximal tubular epithelial cells induced by high glucose in rats.

<正>血管紧张素Ⅱ是一种重要的调节心血管和体液电解质平衡的体液因子。它作用于脉管系统可以引起强有力的收缩效应,刺激肾上腺皮质释放醛固酮,引起渴感和嗜钠,释放血管升压素,调节肾内的管球平衡,反馈抑制肾素的释放。但是血管紧张素Ⅱ在调节体液平衡方面可能有另外的作用,它不仅是一种升高血压的激素,而且是一种在中枢神经系统所产生的神经肽,在中枢神经系统它可能作为一种神经递质或神经功能的调质对渴感,升压素的分泌,肾素的释放,钠的排泄等产生非常重要的作用。

Objective To explore the effect of Rhein on the hypertrophy of renal proximal tubular epithelial cells induced by high glucose and angiotensin II in rats. Methods Studies were performed on anesthetized SD rats. Renal proximal tubular were gained by microdissection and cultured in RPMI-1640 medium. The cell types were identified by immunocytochemistry. The renal proximal tubular epithelial cells were incubated with high glucose (30 mmol/L) and angiotensin II (10(-7) mol/L) to induce the hypertrophy of cells. To observe the effect of Rhein on hypertrophy induced by high glucose and angiotensin II, renal proximal tubular epithelial cells were cultured with different concentrations of Rhein (30, 15, 5 mg/L) for 72 h, then cell size, 3H-leucine incorporation, and cellular protein content were detected to observe the changes. Results High glucose (30 mmol/L) and Ang II (10(-7) mol/L) induced hypertrophy of renal proximal tubular epithelial cells result in, cell size, 3H-leucine incorporation and cellular protein content increased significantly. On the contrary, Rhein inhibited the hypertrophy of renal proximal tubular epithelial cells induced by high glucose and Angiotensin II. Rhein 30 mg/L significantly decreased cell size, 3H-leucine incorporation and cellular protein content. Rhein 15 mg/L decreased 3H-leucine incorporation and cellular protein content. Rhein 5 mg/L decreased cellular protein content. Conclusions Rhein can inhibit the hypertrophy of renal proximal tubular epithelial cells induced by high glucose and Angiotensin II in rats.

目的了解遵义市中学生身体形态发展状况。方法采用分层抽样,于2007年对遵义市1 600名中学生身体形态方面进行调查研究。结果遵义市中学生的身高、体质量、胸围均随年龄增长而增加,但市区各年龄组学生3项指标均值普遍大于郊区。结论遵义市市区中学生身体形态指标(身高、体质量、胸围)高于郊区中学生。