Effect of simulated ischemia-reperfusion on If in sinoatrial node cells and theintervention of KATP channel opener

Objective: To study the effect of simulated ischemia-reperfusion (I/R) on If of sinoatrial node (SAN) cells and the intervention of KATP channel opener Pinacidil. Methods: The SAN cells of the neonatal rats were detached and purified 2 d before the experiment. The experimental animals were randomly divided into the control group, group of simulated I/R, group intervened with KATP channel opener Pinacidil (P+ I/R) and group intervened with KATP channel blocking agent 5-HD (5-HD + P + I/R & 5-HD + I/R). The If density of each group was measured by technique of routine whole cell patch clamp and multiple-catheter perfusion system and the If activated curve in each group was drawn. Results: ①Under different directive potentials, the If density of the SAN cells in I/R group increased significantly, compared with that in the control group ( P < 0.01); that in P + I/R group decreased significantly, compared with that in I/R group ( P < 0.01); the If density values in 5-HD + P + I/R group and 5-HD + I/R group i     

Effects of Chinese Herbal Medicine Serum on the Apoptosis of Sinoatrial Node Cells Induced by Simulated Ischemia-reperfusion

Objective:To study the effect of Chinese herbal medicine Kangxin Fumai Granule(康心复脉颗粒 Granule for heart diseases) serum on the primary cultured sinoatrial node(SAN) cell apoptosis induced by simulated ischemia-reperfusion(IR).Methods:The SAN cells removed from SAN tissue of neonatal Wistar rats were cultured and purified with differential attachment and 5’-bromodeoxyuridine(BrdU) treatment.Simulated IR model was adopted.The obtained cells were morphologically observed with inverted microscopy.By using the method of serum pharmacology,the cell apoptosis was measured with TUNEL staining qualitatively and with flow cytometry quantitatively.Results:Three kinds of cells were observed in the cultured SAN cells:spindle,triangle and irregular.The spindle cells comprised the greatest proportion.The SAN cells in the model group showed moderate positive brown staining in the nucleus,and the apoptosis rate increased significantly compared to that in the control group(P<0.01).While the SAN cells in the Kangxin Fumai Granule high-dose group did not demonstrated positive staining in the nucleus,and the apoptosis rate decreased significantly compared to that in the model group(P<0.05).Conclusion:Of the cells cultured from SAN,the spindle cells were pacemaker cells of SAN in rats.Blockade and/or inhibition of the SAN cell apoptosis might be one of the important mechanisms of Kangxin Fumai Granule in preventing and treating sinoatrial injury induced by simulated IR.     

Primary culture and identification of sinoatrial node cells from newborn rat

Objective To establish a reliable approach to primary culture and identification of sinoatrial node (SAN) cells.Methods The SAN cells were cultured from SAN tissue removed from neonatal Wistar rats and purified with differential attachment and 5’-bromodeoxyuridine (BrdU) treatment. The obtained cells were morphologically observed with inverted microscopy and transmission electron microscopy. Its action potential was recorded using electrophysiological methods.Results Three distinctly different cells were observed in the cultured SAN cells: spindle, triangle and irregular. Of these, the spindle cells comprised the greatest proportion, with their shape, structure and electrophysiological characteristics consistent with those of the pacemaker cells of SAN. The triangle cells were similar in features to the similarly shaped myocytes located in the atrial myocardium.Conclusions The culture method of differential attachment combined with BrdU treatment is a reliable approach to growing SAN cells. Of the cells cultured from SAN, the spindle cells appear to function as pacemaker cells.     

Effects of Propofol on The mRNA Expression of Toll-like Receptor 4 in BV-2 Cells During Mimic Ischemia-Reperfusion Injury In Vitro

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF25) and high-dose propofol (100 μmol/L) intervention group (group PF100). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF100 than in group PF25 (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Effect of Qiangxin Fumai Granule（强心复脉颗粒） on Electrophysiological Functions of the Sinoatrial Node during Ischemia-reperfusion of the Right Coronary Artery in Rabbits

Objective： To study the effect of the Chinese medicine Qiangxin Fumai Granule （强心复脉颗粒, QFG） on electrophysiological functions of the sinoatrial node during ischemia-reperfusion （IR） of the right coronary artery in rabbits. Methods： The right coronary artery IR model in rabbits was adopted. The modeled rabbits were randomly divided into 4 groups： the model group, the atropine group, the highdose QFG group, and the low-dose QFG group, with 8 animals in each group. In addition, twelve rabbits were selected for the sham-operative group. The drugs were administered once via duodenal perfusion after modeling had been stabilized for 10 min. The changes in AA interval, the sinoatrial conduction time （SACT）, the sinus node recovery time （SNRT）, the corrected sinus node recovery time （CSNRT） and the index of sinus node recovery time （ISNRT） at different time points during ischemia and reperfusion were measured. Results： The AA interval was prolonged for more than 40 ms in the model group during ischemia. Compared with the model group, the four electrophysiological parameters abovementioned in the high-dose QFG group and the low-dose QFG group were decreased to different extents at each time point （P〈0.01 or P〈0.05）, and no statistically significant differences were found between the QFG groups and the atropine group （P〉0.05）. Conclusion： QFG is beneficial for accelerating the recovery of sinus node autorhythmicity and conduction function, so as to protect electrophysiological functions of the sinoatrial node. Accelerating the recovery of autorhythmicity and conduction function in the sinus node is considered its electrophysiological mechanism in the treatment of sinoatrial node injury induced by ischemia.     

Effect of mitochondrial KATP channel on voltage-gated K+ channel in 24 hour-hypoxic human pulmonary artery smooth muscle cells

Background Hypoxic pulmonary hypertension (HPH) is initiated by inhibition of O2-sensitive, voltage-gated (Kv) channels in pulmonary arterial smooth muscle cells (PASMCs). The mechanism of hypoxic pulmonary hypertension has not yet been fully elucidated. The mitochondrial ATP-sensitive K+ channel (MitoKATP) is extremely sensitive to hypoxia, and is a decisive factor in the control of mitochondrial membrane potential (ΔΨm). This study investigated the changes of cell membrane potential and Kv channel in cultured human pulmonary artery smooth muscle cell (hPASMC) exposed to 24 hour-hypoxia, and explored the role of MitoKATP and ΔΨm in this condition. Methods Fresh human lung tissues were obtained from the patients undergoing a chest operation. hPASMCs were isolated, cultured, and divided into 6 groups: ① control group, cultured under normoxia; ② diazoxide group, cultured in normoxia with diazoxide, an opener of MitoKATP; ③ 5-HD group, cultured in normoxia with sodium 5-hydroxydecanoate (5-HD), an antagonist of MitoKATP; ④ 24 hour-hypoxia group; ⑤ 24 hour-hypoxia + diazoxide group; and ⑥ 24 hour-hypoxia + 5HD group. Whole-cell patch-clamp technique was used to trace the cell membrane K+ currents. The expressions of cell membrane Kv1.5 mRNA and protein were determined by RT-PCR and Western blot technique, respectively. The relative changes in mitochondrial potential were tested with rhodamine fluorescence (R-123) technique. Results After exposure to diazoxide for 24 hours, the intensity of R-123 fluorescence in normoxic hPASMCs was significantly increased compared with control group (P&lt;0.05), but there were no significant changes in these tests after the hPASMCs had been exposed to 5-HD for 24 hours. Twenty-four hour-hypoxia or 24 hour-hypoxia + diazoxide could markedly increase the intensity of R-123 fluorescence in hPASMC and the changes were more significant in 24 hour-hypoxia +diazoxide group than in 24 hour-hypoxia group (P&lt;0.05) although 5-HD could partly weaken the effect of 24 hour-hypoxia on the intensity of R-123 fluorescence. After exposure to diazoxide for 24 hours, the cell membrane K+ currents and the expression of cell membrane Kv1.5 mRNA and protein in normoxic hPASMCs were significantly decreased compared with control group (P&lt;0.05), but there were no significant changes in these tests after the hPASMCs had been exposed to 5-HD for 24 hours. Also, 24 hour-hypoxia or 24 hour-hypoxia + diazoxide decreased the cell membrane K+ currents and the expression of Kv1.5 mRNA and protein (P&lt;0.05) but the changes were more significant in 24 hour-hypoxia + diazoxide group than in 24 hour-hypoxia group (P&lt;0.05). Again, 5-HD could partly weaken the inhibitory effect of 24 hour-hypoxia on the cell membrane K+ currents and the expression of Kv1.5 mRNA or protein (P&lt;0.05). Conclusions The opening of MitoKATP followed by a depolarization of ΔΨm in hypoxia might contribute to the alterations in the expression of cell membrane Kv1.5 mRNA and protein leading to change in the cell membrane potential of hypoxic hPASMCs. This might be a mechanism of the development of hypoxic pulmonary hypertension.     

Effects of propofol on the mRNA expression of toll-like receptor 4 in BV-2 cells during mimic ischemia-reperfusion injury <Emphasis Type="Italic">in vitro</Emphasis>

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 （TLR4） in BV-2 cells during mimic ischemia-reperfusion （I/R） injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random： control group （group C）, ischemia/reperfusion group （group I/R）, low-dose propofol （25 μmol/L） intervention group （group PF25） and high-dose propofol （100 μmol/L） intervention group （group PF100）. The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C （P〈0.01）. And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C （P〈0.01）. After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R （P〈0.01）. And the decrease in those indicators was more significant in group PF100 than in group PF25 （P〈0.01）. It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

The influence of insulin on secretion of IGF-Ⅰ and IGFBP-Ⅰ in cultures of human endometrial stromal cells

Objectives To study the influence of insulin on IGF-Ⅰ and IGFBP-Ⅰ secretion of the human endometrial stromal cells. Methods Late proliferative phase endometrial stromal cells were isolated from endometrium tissues and then cultured for 24 h in Hams F-12 only as a control and in Hams F-12 with different concentrations of estradiol (E2) and insulin (INS) as treated groups. Simultaneously, the endometrial stromal cells from late secretory phase endometrium were cultured for 24 h in Hams F-12 only as a control and in Hams F-12 supplemented with different concentrations of progesterone (P) and insulin as treated groups. After 24 h of culturing, the mediums were collected for either IGF-Ⅰ or IGFBP-Ⅰ assays. Result The concentrations of IGF-Ⅰ in medium from cultured endometrial stromal cells in the proliferative phase were 0.78±0.47 ng/ml in the hormone-free control group; 1.44±0.59 ng/ml and 1.39± 0.33 ng/ml in 100 pg/ml E2 group and 20 μU/ml INS group, which was higher than that of the control group (P&lt;0.05 and P&lt;0.01, respectively). The IGF-Ⅰ concentration in the 100 μU/ml INS group was 2.03±0.53 ng/ml, which was higher than that of the 20 μU/ml INS group (P&lt;0.01). Levels of IGF-Ⅰ in the 100 pg/ml E2 plus 20 μU/ml INS group was 2.18±0.36 ng/ml, which was significantly higher than that of the 20 μU/ml INS and 100 pg/ml E2 group (P&lt;0.01), but lower than that of the 100 pg/ml E2 plus 100 μU/ml INS group (3.42±0.75 ng/ml), P&lt;0.01. The concentration of IGFBP-Ⅰ in medium from cultured endometrial stromal cells in the secretory phase was 2.50±1.39 ng/ml in the hormone-free control group and 5.44±2.09 ng/ml in the 10 pg/ml P group, which was significantly higher than that of the control (P&lt;0.01). IGFBP-Ⅰ concentration in 20 μU/ml INS group was 0.16±0.58 ng/ml, which was lower compared with control, but higher compared with the 100 μU/ml INS group (P&lt;0.01). The level of IGFBP-Ⅰ in the 10 ng/ml P plus 20 μU/ml INS group was 2.10±1.17 ng/ml, lower compared with the 10 ng/ml P group, but higher compared with the 10 pg/ml P plus 100 μU/ml INS group, P&lt;0.01. Conclusions Insulin can stimulate basal (without hormone) and E2-stimulated IGF-Ⅰ secretion in cultured stromal cells from human late proliferative endometrium in a dose-dependent manner. Insulin can suppress basal (without hormone) and P-stimulated IGFBP-Ⅰ secretions in cultured stromal cells from human secretory endometrium in a dose-dependent manner.     

The antiapoptotic effect of insulin against anoxia/reoxygenation injury in cultured cardiomyocyte of neonatal rat

Objective： To study protective effect of insulin against cardiomyocyte apoptosis in anoxia/reoxygenation （A/R） injury of neonatal rat. Methods： The model of A/R injury was finished through receiving anoxia for 2h and reoxygenation for 4h in cultured cardiomyocytes of neonatal rat. The cardiomyocytes were divided randomly into 3 groups： control group （CON）, anoxia/reoxygenation group （A/R） and insulin-treated group （INS）. At the end of reoxygenation of 4 hours, activities of lactate dehydrogenase （LDH）, contents of malondiaidehyde （MDA）, were assessed through spectrophotometric procedures, myocyte apoptosis were detected through TUNEL and DNA Ladder. Results： MDA, LDH, and Apoptosis Index were significantly decreased in INS group compared with A/R group （P〈0.01）. Conclusion： Insulin has a protective effect against A/R injury in cultured cardiomyocyte of neonatal rat; the protective mechanism may contribute to antiapoptosis of insulin.     

麻疹的双相移位研究进展

近年新生儿、婴儿、成人麻疹患者逐年增加,临床表现一般仍较典型,成年人麻疹患者全身中毒症状较重。麻疹抗体检测结果阳性是主要的诊断依据。麻疹发病的双相移位的机理可能是,免疫保护力不足,婴儿出生时麻疹抗体力低。孕期母传胎的麻疹抗体减弱,母经乳汁传给婴儿的抗体减弱,成人麻疹抗体水平逐年下降。预防措施是怀孕前给予育龄妇女麻疹疫苗接种,鼓励母乳喂养,麻疹疫苗计划免疫适当提前,在成人追加麻疹疫苗的免疫,加强病毒变异的研究等。     

87株幽门螺旋杆菌VacA等位基因的分析

目的以研究方法LiPA（Line Probe Assay）分析VacA等位基因的表达,了解幽门螺旋杆菌致病机理的理解。方法从三个不同城市87位进行胃镜检查患者的胃粘膜中培养出幽门螺旋杆菌,提取DNA,用LiPA方法分析VacA等位基因。结果（1）87位患者以sic（88．5％）和m2a（63．2％）分布为主,未发现s1b和s2;（2）混合菌株感染率为41．4％,远远高于西方国家,其中上海的混合感染率最高（62．5％）,与北京（41．0％）和南宁（20．8％）相比具有显著性差异,P〈0．01;（3）北京、海、南京三个不同城市s1、m等位基因亚型分布率存在差异;（4）溃疡病和非溃疡病患者m1和m2的分布率没有显著性差异。结论我国幽门螺旋杆菌多重菌株感染率较高,不同的m基因型与消化性溃疡的发生无显著性相关。     

尿液pH值对红细胞检验影响的探讨

[目的 ]通过尿液 pH值对红细胞检验影响的观察 ,更加科学、准确地诊断血尿和血红蛋白尿。[方法 ]采用干化学分析仪检测和尿液显微镜红细胞计数 ,观察 180例正常人尿标本加入正常人血标本后 ,不同 pH值 ,不同时间内 ,观察红细胞溶解情况。 [结果 ]pH <5 .5以下时 ,随着时间的延长 ,红细胞溶解现象明显。 1h后观察有显著性差异 (P <0 .0 5 ) ;2h后有非常显著性差异 (P <0 .0 1)。[结论 ]pH <5 .5时对红细胞计数影响较大 ,易致红细胞发生溶解现象 ,出现假性血红蛋白尿 ,对血尿和血红蛋白尿很难区分 ,给临床诊断造成不便 ,更易引起漏诊和误诊。     

阴道炎1236例病原检查分析

对１２３６例阴道炎患者的阴道分泌物作直接镜检和病原菌分离培养检查；结果，细菌感染９００例，念珠菌２３４例，滴虫１０２例。９００例细菌经鉴定；葡萄球菌３００例，阴道加特纳菌２７６例，淋病奈瑟菌１７０例，其它细菌１２４例。结果表明，葡萄球菌，阴道加特纳菌，淋病奈瑟菌是细菌性阴道炎最常见的致病菌。     

喉癌主癌灶手术切缘的临床特点分析

目的探讨研究喉癌主癌灶手术的安全切缘在临床上的特点。方法回顾性分析2007年6月—2011年3月已经确诊喉癌的患者100例,随机分成A、B两组,A组为观察组50例,主要采取手术切缘后,然后采取镜下观察分析;B组为观察组50例,主要采取手术切缘后,然后采取肉眼观察分析,将结果进行临床特点分析比较。结果早期的喉癌患者和晚期的喉癌患者的阳性切缘例数,差异无统计学意义（P〉0.05）;晚期的阳性切缘发生次数高于早期的阳性切缘发生次数,差异有统计学意义（P〈0.05）。声门上区[SG]2、3和5、10mm;跨声门型[TG]2、3mm和5、10mm的切缘对比,差异有统计学意义（P〈0.05）。SG、G、TG、IG的2mm和3mm,5mm和10mm对,差异无统计学意义（P〉0.05）。肉眼阳性切缘观察39个,镜下阳性切缘观察43,两者差异无统计学意义（P〉0.05）。结论依据原发不同部位、不同分期和不同范围选取适合的切线,就可以有效地减少阳性切缘的发生。     

碱量法测定壳多糖脱乙酰度的方法探讨

目的：评价壳多糖脱乙酰度测定的减量法的影响因素。方法：通过实验评价碱量法的精准度及样本含潮量等因素对测定结果的影响,并对汪氏推荐计算公式和本文作者提出的改良公式作出评价。结果：碱量法测定壳多糖脱乙酰度受样本性状、含潮率、反应体系中酸量的影响。改良公式更能反映样本实际脱乙酰度。结论：碱量法简便易行,评价指标在可接受范围,可用于壳多糖研制中的质量评价。     

上颌骨牙源性囊肿刮除手术的疗效观察

目的探讨对上颌骨牙源性囊肿患者进行囊肿彻底刮除手术的临床疗效。方法对我科在2010年1月～2017年9月收治的73例上颌骨牙源性囊肿患者行囊肿彻底刮除手术治疗,对患者术区肿胀消退、术后伤口感染、伤口愈合、牙龈再附着、术后复发、骨质改建、骨质修复等情况随访观察。结果 73例患者术后肿胀消退时间为1～4 d。73例患者术后均未发生伤口感染,伤口均一期愈合。所有患者牙龈再附着情况好,术后均未见复发。术后未见并发症。骨质改建效果好,骨质修复的效果因影像学资料过少,缺乏客观依据,暂不下有效结论。结论对上颌骨牙源性囊肿患者进行囊肿彻底刮除手术,术后患者的恢复情况良好,值得在临床治疗上进行推广。     

儿童腹痛328例病因诊断及分析

目的：根据儿童急性腹痛的特点，诊断儿童腹痛应综合临床及相关检查提供的依据，做出准确诊断，给予适当治疗。方法：对临床328例腹痛患儿进行选择性辅助检查和分析。结果：小儿腹痛病因复杂，腹腔内疾病占64．63％，共17种病因；腹腔外疾病占35．37％，共6种病因。结论：仔细询问病史、认真体格检查、密切观察病情、针对性地选择辅助检查，在儿童腹痛的诊断上具有重要的意义。     

手术治疗髋臼骨折45例效果观察

目的:探讨髋臼骨折患者的手术治疗效果。方法:系统性回顾收治的45例髋臼骨折患者手术治疗效果。结果:本组患者随访2～8年,治疗的总优、良患者38例,总优良率达到84%,手术切口全部愈合,未发生切口感染及深部感染,在患者中5例出现并发症:1例坐骨神经损伤,1例股骨头坏死,1例异位骨化,2例创伤性关节炎。结论:充分的术前计划,恰当的手术时机,正确的入路选择以及良好的术后锻炼,是治疗髋臼骨折的有效方法。     

癫痫常见临床误诊原因分析

目的通过对癫痫和非癫痫误诊病例及24小时脑电图确诊癫痫患者的临床特点分析,提高癫痫的诊断率,减少癫痫误诊。方法纳入四川大学华西医院2009年10月～2011年4月收治的误诊病例及经脑电图确诊的癫痫患者共172例,分析其误诊原因和临床特点。结果在纳入的172例患者中99例非痫性发作误诊为癫痫发作,43例癫痫发作误诊为非痫性发作,30例经24小时脑电图确诊为癫痫。癫痫最易被误诊为短暂性脑缺血发作(TIA)、晕厥、假性发作。反之,非癫痫发作误诊癫痫发作多见于假性发作、TIA。对24小时脑电图确诊的癫痫发作患者的临床特点分析发现不自主咂嘴合并吞咽动作、上腹部不适是最常见待诊患者的临床表现。结论熟练掌握癫痫与非癫痫发作性疾病的临床表现,仔细询问病史,认真做好体格检查,结合脑电图尤其是24小时视频脑电图等检查进行综合分析,可以减少癫痫的误诊,提高癫痫的诊断率。