医学课程体系的整合是培养21世纪复合型医学人才的必由之路.2013年以来,安徽医科大学开始了五年制临床医学专业整合课程的教学改革试点,其中生物化学与分子生物学和细胞生物学被整合为生命的化学.整合课程在实施过程中组织了跨学科教学团队,在原有生物化学和细胞生物学整合的基础上添加了原生理学中细胞的生物电专业知识,创新了教学内容,贯彻了以学生为中心和自主学习的教学理念;逐渐摸索建立了形成性评价体系进行课程考核和成绩评定,取得了较好的教学效果.

AIM:The present study examined the hyperpolarizing effects of endothelium derived hyperpolarizing factor (EDHF) / hydrogen sulphide (H2S) on the membrane potential of the nerve cell in mice.METHODS:In presence of 3 × 10-5 mol/L N-nitro-L-arginine-methyl-ester (L-NAME,an inhibitor of nitric oxide synthase) plus 10-5mol/L indomethacin (Indo,an inhibitor of PGI2 synthsis),to evaluate the hyperpolarizing effect of acetylcholine (ACh,10-5.5-10-8 mol/L) on nerve cell in CSE+/+ and CSE knockout (CSE-/-) mice and NarHS (10-5-10-2.5 mol/L) on the nerve cell in CSE+/+ mice.RESULTS:ACh (10-8-10-5.5 mol/L) induced concentration-dependent hyperpolarization nerve cell in CSE+/+ mice while Significantly reduced by remove MCA/Endo or PSS replace.The non NO/PGI2 responses induced by ACh were abolished by the co-application of 10-7 mol/L charybdotoxin (ChTX) and 5 × 10-6 mol/L apamin (Apa)which blocked intermediate-and small-conductane calcium-activated potassium (IKca and SKca) channels.The endogenous H2S synthase inhibitor 10-4 mol/L DL-propargylglycine (PPG) or CSE-/-were also abolished the effect.The H2 S donor NarHS (10-5-10-2.5 mol/L) elicited concentration-dependent hyperpolarization nerve cell in CSE +/+ mice and was inhibited by 10-7 mol/L Iberiotoxin (IBTX) which blocked large-conductane Kca (BKca) channels.CONCLUSION:H2S act as EDHF causes nerve cell hyperpolarization.

Objective To record the myocardial cell bioelectric activities and explore electrophysiological profiles of Gastrodin on rat myocardial cells. Methods Sinoatrial node cells and atrial myocytes isolated from 50 SD rats were used in this study to characterize the effect of Gastrodin on their electrophysiological profiles with intracellular recording approaches. Results In the present experimental conditions, the bioelectric activities of sinoatrial node and atrial myocytes were truly recorded and we found that Gastrodin significantly accelerated AP frequency and Vmax 4 of sinoatrial node cells and shortened APD90 of atrial myocytes. Conclusion The experimental conditions of recording the bioelectric activities of myocardial cells in vitro are relatively mature. Gastrodin can affect the elec-trical activity of myocardial cells in vitro and the effects of Gastrodin may be related to inhance the permeability of T-Ca2+ channels and promote the opening of KATP.

Objective To observe the possible mechanism of Gastrodin precondition on attenuating reaction of myo-cardial ischemia reperfusion injury ( MIRI) in rats. Methods Sixty SD rats were randomized into five groups such as Sham group, MIRI group and Gastrodin low-dose(0. 1 g/kg),middle-dose (0. 2 g/kg) and high-dose (0. 4 g/kg) groups. Then the rats mode of myocardial I/R injury were prepared by closing the left anterior descending cor-onary artery for 30 mins and then releasing it for 2 hrs under anesthesia. The changes of tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6) in blood serum and myocardial tissue were determined, and sareoplasmic retic-ulum Ca2+-ATP ( SERCA) was determined in myocardial tissue. Results Compared with MIRI group, the TNF-αand IL-6 of blood serum and myocardial tissue were lower ( P<0. 05 ) and the concentration of SERCA of myocar-dial tissue was higher ( P<0. 01 ) in Gastrodin group of middle and high concentrations. Conclusion Gastrodin preconditioning can attenuate myocardium ischemia reperfusion injury in rats which maybe related to its anti-inflam-mation function and its increase of SERCA.

OBJECTIVE: To observe the effect of gastrodin preconditioning on the myocardial ischemia reperfusion injury (MIRI) in rats and to study the mechanism. METHODS: SD rats were randomized into six groups including sham group, NS group, gastrodin groups of low middle and high concentrations and verapamil group. The rats were treated with gastrodin via intragastric administration for one week. Then the rats were treated by closing the left anterior descending coronary artery for 30 min and then releasing it for 2 h under anesthesia. The changes of the arrhythmia, myocardial histological structure and serological tests including SOD, LDH, CK-MB and MDA were examined. Furthermore, the effect of gastrodin on store-operated Ca2+ influx was also investigated. RESULTS: It was found that gastrodin significantly enhanced SOD production and suppressed the incidences of arrhythmia and LDH, CK-MB and MDA concentrations. Additionally, gastrodin inhibited store-operated Ca2+ influx. CONCLUSION: Gastrodin has protective effect on myocardium against ischemia reperfusion injury in rats through several pathways including cleaning oxygen radicals and reducing the overload of Ca2+.

Objective To study the protective effects of Ginkgolide B(GB) on celiac ganglion(CG) neurons induced by intestinal ischemia/reperfusion in guinea pig in vitro.Methods 30 guinea pigs were randomly divided into three groups: control group,ischemia/reperfusion(I/R) group,I/R+GB group(treated with GB 2.5×10-4 mol/L).The intestinal injury was induced by clamping the superiormesenteric artery for 30 min and reperfusion for 60 min.The intracellular bioelectric recording technique and HE staining were applied to monitor the protective effects of GB on celiac ganglion neurons.Results In this study,CG neurons of the ischemia/reperfusion group were injuried. The intracellular bioelectric recordings showed GB could improve and recovery some physiologic function of CG neurons,stabilized the potential of membrane.With the stimulation of depolarized square-wave,the action potential was observed in 70.1% (61/87) cells of normal group compared with 12%(3/25) cells of I/R group.Under the same condition,in I/R+GB group,42.4% (20/48) cells depolarized and the action potential was found in 31.2%(15/48) cells(P<0.05).In HE staining observation,after the perfusion with GB,the amount of CG neurons of the group was much more than that of I/R group (P<0.05).Conclusion GB is responsible for the protective effect on CG neurons which are induced by intestinal ischemia/reperfusion in guinea pigs in vitro.

Objective To investage the effect of extract of ginkgo biloba(EGb) on gastric motility in different states and discuss the mechanism.Methods Using blue dextran 2000(DB 2000) as marker,we observed the effect of EGb on gastric emptying rate.And the content of NO in bllod plasma and stomach muscle layer were measured by chemical method. Results EGb had no effect on gastric motility in normal rat.cold-restraint stress inhibited significantly gastric emptying(P0.05),and EGb attenated significantly stress-induced gastric emptying rate decrease(P0.05).In the stress model group,NO content in blood plasma decrease markedly by EGb(P0.01).Conclusion EGb can mediate gastric discord by cold restraint stress,NO might take part in the effect.

The present study was carried out to determine the regulation of ginkgolide B(GB)on fast-excitatory postsynaptic potential(fast-EPSP)in the neurons of celiac ganglion(CG)and the mechanisms involved in the GB-induced responses.Intracellular recording was employed to measure the fast-EPSP in the neurons of CG freshly isolated from Guinea Pigs(Cavia porcellus).We found that perfusion of GB(4×10-6 mol/L)significantly decreased the amplitude of fast-EPSP in the CG neurons(n=12,P<0.05).The fast-EPSP was completely inhibited by a low Ca 2+/high Mg 2+ Krebs solution(n=3),but significantly enhanced by a high Ca 2+ Krebs solution(n=12,P<0.05),suggesting a contribution of extracellular Ca 2+ to the generation of the fast-EPSP.The high Ca 2+ Krebs solution-induced augmentation of the fast-EPSP was significantly attenuated by GB(n=12,P<0.05).These results indicate that GB negatively regulates fast-EPSP in CG neurons in Guinea pigs through an inhibition on external Ca 2+ entry.

The in vitro spontaneous fast-excitatory postsynaptic potential(f-EPSP)and spontaneous action potential(AP)were recorded in the celiac ganglion(CG)neurons of Guinea-pig by the intracellular recording technique and the related possible mechanism was analyzed.The spontaneous f-EPSP and AP had different frequencies in the CG neurons.Amplitudes of f-EPSP(13.26±6.74)mV(n=34)and spontaneous f-EPSP(5.67±2.66)mV(n=26)were significantly different(P<0.01).Amplitudes of AHP(8.99±2.79)mV(n=54)and spontaneous AHP(13.86±4.24)mV(n=30)were also significantly different(P<0.01).Spontaneous AP could be blocked by hexamethonium or low Ca 2+/high Mg 2+ Krebs solution,whereas spontaneous f-EPSP could be partly blocked by the same treatment.The results indicate that spontaneous bioelectricity exists in isolated CG neurons,and this may be induced by ACh and other calcium-independent neurotransmitters.

Aim To investigate the electrophysiological character of normal,ischemic and reperfusion of ventricular myocardium in vitro in guinea pig by means of computer signal acquisition and processing technique.Methods Superfusion of ventricular myocardium in vitro and intracellular recording technique were adopted.Results The mean value of resting potential(RP) of ventricular myocardium of guinea pig was(-72±6.58)mV.The mean value of action potential amplitude(APA) was 112.22±8.65 mV.The maximum of depolarization velocity(Vmax) was 107.37±10.55 mV·s-1.The action potential duration at 10%,50% and 90%(APD10,APD50 and APD90) was 39.25±8.55 ms,123.45±15.25 ms and 169.35±18.12 ms respectively.After superfusion by ischemia and hypoxia Tyroad solution,the APA and Vmax decreased and APD90 shortened significantly.After reperfusion 2 minutes,The Vmax continuing to decrease significantly showed that the effects of ischemic aggravated.After reperfusion 10 minutes there was RP and AP reablement on the whole.Conclusion As for research of electrophysiological character of ventricular myocardium,the computer signal acquisition and processing technique could be used as a powerful tool to obtain objective and real data for analyzing the mechanism of changing and formation of cell bioelectricity or investigating more about effects of pathology and drug on electrophysiological properties of myocardium.

AIM To study whether store-operated Ca2+ channel (SOC) is present in rat colonic smooth muscle cells. METHODS Intracellular Ca2+ ([Ca2+]i) changes induced by thapsigargin- or caffeine-activated SOC channel were measured in enzymatically dissociated rat colonic smooth muscle cells with the fluorescent indicator Fura-2/AM. RESULTS In the absence of external Ca2+ , the sarco-endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 micromol/L) and ryanodine receptor (RyR) activator caffeine both transiently elevated [Ca2+]i from (68.32 +/- 3.43) nmol/L to (240.85 +/- 12.65 ) nmol/L, (481.25 +/- 34.77) nmol/L. A subsequent reintroduction of Ca2+ into the extracellular solution resulted in [Ca2+]i further elevated to (457.55 +/- 19.80) nmol/L, (1005.93 +/- 54.62) nmol/L; (643.88 +/- 34.65) nmol/L, (920.16 +/- 43.25) nmol/L, respectively. And the elevated response was blocked by lanthanum (1 mmol/L), but was insensitive to L-type voltage calcium channels blocker verapamil and membrane depolarization. CONCLUSION SOC activated by store depletion are present in rat colonic smooth muscle cells. And we further prove the existence of such Ca2+ channels in excitable cells.

Objective To investigate and analyze the effect of the total flavone ginkgo (TFG) on membrane potential of the neurons in celiac ganglia (CG) of guinea-pig in vitro. Methods Intracellular microelectrode recording and administration of superfusion were used. Results Superfusion of celiac ganglia with TFG evoked a concentration-dependent membrane depolarization in 45.8% of the neurons, amplitude and duration are 7.9 mV±0.7 mV and 245.9 s±29.2 s respectively and 10% demonstrated hyperpolarization and 10% biphasic response which consisted of an initial hyperpolarization phase and a followed depolarization, also 34% no response to TFG. The depolarization of most neurons could be blocked partly by cholinoceptor (atropine and hexamethonium) and adrenoceptor antagonist (phentolamin and propranolol); Among the depolarization neurons, the membrane resistance (Rm) decreased in 47.2% increased in 21.8% and unchanged in 30.9%; The depolarization can be blocked partly by Na~+ and Ca~ 2+ free Krebs solution and depolarization increased by traethylammonium (TEA). Conclusion TFG can evoke the depolarization of most CG neurons, and the mechanism probably is associated with the increasing influx of Na~+ and Ca~ 2+ and the decreasing outflow of K~+.

Objective To investigate the effect and mechanism of total ginkgo flavonoids(FG) on myocardial injury in diabetic rats. Methods Rats were given streptozotocin(STZ) to induce the diabetic rats model. The maleic dialdehyde (MDA), creatine phosphokinase(CK), nitric oxide(NO) contents and the activity of superoxide dismutase (SOD), lactate dehydrogenase (LDH) and nitric oxide synthase (NOS) in myocardium were measured by spectrophotometric method with different dose of FG. The ultrastucture of myocardial cells was detected with eletronmicroscope. Results FG 100, 200 mg/(kg·d) markedly inhibited the increases of the content of MDA and the decreases of the content of CK in mice myocardial tissue. FG 50, 100, 200 mg/(kg·d) increased the activity of SOD and markedly inhibited the reduction of NO content and the activity of NOS in rats myocardium, FG 200 mg/(kg·d) increased the activity of LDH. Electron microscopic observation of myocardial muscle of the DM rats revaled typical alterations consisting of cutaneous or interruption of the myocardial muscle fiber, absence of the normal structure of the sarcomere, swell of the mitochondria and disappearance in some transverse striation. FG 200 mg/(kg·d) treatment group could greatly improve the above changes. Conclusion FG treatment has a protective effect on myocardial injury in diabetic rats, and the mechanism may be via attenuating oxygen free radicals and increasing NO production and decreasing the ultrastucture of myocardial cells.

Objective To investigate and analyze the effect of the total flavonoids of ginkgo (TFG) on the late slow excitatory postsynaptic potential (ls-EPSP) of the neurons of inferior mesenteric ganglion (IMG) of guinea pig in vitro. Methods intracellular microelectrode recording and administration of superfusion.Results The mean value of resting potential (RP) of neurons of IMG of guinea pig was (-53.1±6.5)mV(n=54). Repetitive stimulation(10 V,1 ms,20 Hz,4 s)of hypogastric nerves (HN) elicited a burst of action potentials which was followed by ls-EPSP in 61.1% (33/54) of the IMG neurons. The amplitude and duration of ls-EPSP were (7.7±1.8)mV and (97.4±17.4)s(n=33),respectively. Superfusion of IMG with TFG evoked a concentration-dependent depression [FQ(11。46,X-WZ][CD44]on the amplitude and duration of ls-EPSP in 69.7% of the neurons and 21.2% had no response to TFG. The depression induced by TFG was not blocked by cholinoceptor and adrenoceptor antagonists, but was eliminated in a low Ca~ 2+ /high Mg~ 2+ Krebs solution. The depression induced by TFG could be enhanced by low Na~+(58 mmol/L) or high K~+(20 mmol/L) Krebs solution. It was also enhanced by TTX (10 μmmol/L)and weaken by tetraethylammonium(TEA,1 mmol/L). Conclusion The inhibition of TFG on ls-EPSP could be observed on most neurons of IMG of guinea pig and the mechanism probably is associated with the decreasing influx of Na~+ and increasing outflow of K~+.

Contraction of smooth muscle cells is triggered by an increase in cytosolic Ca(2+) upon agonist stimulation. Ca(2+) influx across the plasma membrane constitutes a major component of the agonist-induced response in smooth muscle cells. Traditionally, voltage-operated Ca(2+) channel (VOCC) is considered as the channel mediating the Ca(2+) entry. However, this view has been challenged by recent discoveries, which demonstrated that other types of ion channels, such as store-operated and/or receptor-operated Ca(2+) channels (SOCC and/or ROCC), also participate in Ca(2+) response induced by agonists in smooth muscle cells. SOCC is defined as the channel activated in response to the depletion of the internal Ca(2+) stores, an event secondary to G protein coupled receptor or receptor tyrosine kinase stimulation. The Ca(2+) flow mediated by SOCC is termed as capacitative Ca(2+) entry (CCE). Previous study from other group has demonstrated that VOCC played a predominant role in ACh-induced contraction of distal colon smooth muscle in guinea pig. However, whether SOCC participates in the agonist-induced contractile response in this particular tissue is unknown. The present study was performed to investigate the role of CCE in ACh-induced mechanical activity of distal colon smooth muscle in rats. The contractile function of the smooth muscle was assessed by measuring isometric force of isolated rat distal colon rings. We showed that both high extracellular K(+) (40 mmol/L) and ACh (5 mumol/L) evoked striking contraction of the smooth muscle. The contractile responses were almost abolished by removal of extracellular Ca(2+) with ethylene glycol-bis(2-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA), suggesting a critical contribution of extracellular source of Ca(2+) to the contraction. Verapamil (5 mumol/L), an L-type VOCC blocker, significantly attenuated, but didn't completely eliminate the high K(+)- and ACh-induced contraction (74% and 41% for high K(+) and ACh, respectively), indicating that additional channels might be involved in the contractile mechanism. Furthermore, ACh only induced transient contractions in the absence of extracellular Ca(2+). Readmission of Ca(2+) into the extracellular compartment resulted in a significant and sustained increase in the tension of the smooth muscle. This response was not affected by verapamil (5 mumol/L) and Cd(2+) (5 mumol/L), both of which efficiently block VOCC at the doses. However, La(3+), a known inhibitor of SOCC, significantly suppressed the Ca(2+) readdition-induced contraction in a dose-dependent manner. On the basis of these results, we conclude that contraction of smooth muscle in the distal colon is regulated by multiple Ca(2+) channels. In addition to VOCC-mediated Ca(2+) influx, SOCC-mediated CCE participates in agonist-induced contractile response of distal colon smooth muscle in rats.

目的应用计算机信号采集和处理技术系统地研究豚鼠离体腹腔神经节细胞电生理特性.方法离体神经节灌流,细胞内微电极记录技术.结果豚鼠CG细胞静息电位(RP)为(-62.26±11.27) mV (n=110),膜电阻(Rm)为(79.17±27.16) MΩ,膜电导(G) 为(14.40±5.60) nS (n=63),膜电容(Cm)为(231.53±105.24) pF,膜时间常数(τ) 为(2.01±1.07) ms (n=36);直接胞内刺激引起的锋电位幅度(APA)和时程(APD)分别为(63.41±12.18) mV和(2.12±0.46) ms,锋电位后超极化电位幅度(AHPA) 和时程(AHPD)分别为(16.37±3.72) mV和(16.1±5.24) ms (n=75);刺激内脏大神经引起的快兴奋性突触后电位(f-EPSP)幅度和时程分别为(13.26±6.74) mV和(45.20±10.6) ms (n=34),刺激内脏大神经引起的APA 和APD分别为(69.71±15.38) mV和(2.13±0.55) ms (n=54),AHPA和 AHPD分别为(8.58±2.77) mV和(12.8±0.85) ms (n=48).结论计算机信号采集和处理技术在细胞电生理研究方面的应用,为全面地分析细胞生物电的变化及其形成机制,亦为深入研究递质、调质以及某些药物对细胞生物电的影响提供真实客观的数据.

钙库操纵性通道(SOC)是目前研究较热门的一种离子通道,其开放与关闭受内质网中Ca2+贮量调控.SOC参与机体许多重要生理功能的调节,尤其在平滑肌紧张性变化的调节中起重要作用.果蝇瞬时受体电位(transient receptor potential,TRP)蛋白在光信号传递中发挥重要作用,在哺乳动物中,发现TRP蛋白的同系物TRPC1蛋白是SOC的组成成分.研究并深入了解SOC的特性对于开发一类新的钙通道拮抗剂具有重要的理论意义.

Aim To investigate effects of ginkgo biloba extract (EGb) on the change of gastrointestinal motility and NO content induced by cold restraint stress. Methods Using blue dextran 2000 (DB-2000) as marker, we observed the effects of EGb and stress on gastric emptying and motor activity of small intestine. And the content of NO in plasma and gastrointestinal muscle layer were measured by chemical method. Results Cold-restraint stress can inhibit significantly gastric emptying and motor activity of small intestine in rats,and by contrast EGb can reverse these effects evoked by cold-restraint stress. In the stress groups in the presence of EGb, NO concentrations in plasma makedly decreased compared with the control groups in the absence of EGb. Conclusion EGb can mediate directly gastrointestinal discord by cold restraint stress, NO might take part in the effect.

目的探讨银杏叶提取物(EGb)对豚鼠离体结肠运动的影响及其机制.方法制备豚鼠离体结肠肠段标本,通过灌流给药的方法观察不同浓度EGb、银杏黄酮等对离体结肠收缩活动的影响.结果 EGb (0.5～50.0 mg/L)可对结肠自主活动产生抑制和兴奋两种不同的影响,其中以抑制作用为主.EGb也可部分拮抗由ACh、5-HT引起的结肠运动的增强效应;并协同血管活性肠肽(VIP)、无钙Kreb液灌流引起的抑制作用;对γ-氨基丁酸、肾上腺素引起的结肠运动抑制效应无显著影响;用M受体阻断剂阿托品预处理可部分阻断EGb对结肠自主活动的影响.银杏黄酮(0.01～0.1 mg/L)可使豚鼠离体结肠运动抑制,且表现一定的剂量依赖性.结论 EGb对豚鼠结肠运动的影响可能是通过影响5-HT、ACh、VIP等肠神经系统递质的释放,经M受体介导以及对平滑肌的直接作用等多种途径实现的.银杏黄酮可能是EGb发挥抑制作用的有效部位.

钾离子通道是人体神经元、心肌细胞上分布最为密集、功能最为重要的一种通道,对于细胞膜兴奋性的维持、动作电位的延缓、成簇放电的频率、神经网络的学习记忆储存等具有重要作用.