肉毒毒素A对大鼠肛门括约肌作用的实验研究

目的了解肉毒毒素A对肛门括约肌的作用效果,并初步探讨其作用机制。为肉毒毒素A治疗肛直肠痉挛性疾病奠定理论基础。方法取Wistar大鼠75只,随机分为3组,每组25只。分别注射肉毒毒素A、生理盐水和阿托品溶液。在注药前、后分别检测3组动物的肛周括约肌肌电图,同时行肛直肠测压。此外,作肛门括约肌的病理组织切片和苏木素-伊红(HE)染色,并在光镜下观察其组织学形态。结果治疗前3组动物间肌电图表现差异无统计学意义(P>0.05),平均电位振幅(97±36)μV,平均时限(3.8±0.9)ms。治疗后72h,肉毒素A治疗组的肌电图运动电位的平均振幅为(68±29)μV,低于阿托品组和生理盐水组;平均时限(4.6±1.2)ms,长于阿托品组和生理盐水组,差异有统计学意义(P<0.05)。肉毒毒素治疗组的肛直肠静息压、肛直肠最大自主收缩压、直肠肛门抑制反射值均低于生理盐水组和阿托品组,差异有统计学意义(P<0.05)。结论肉毒毒素A能有效降低肛门括约肌的压力,为临床治疗慢性出口梗阻型便秘疾病提供了新的方法和思路,值得进一步研究。     

关节腔局部注射不同剂量氨甲环酸对单侧全膝关节置换术失血的影响

目的 探究关节腔局部注射不同剂量氨甲环酸(TXA)对减少单侧全膝关节置换术(TKA)失血的影响。方法 收集2013年1月至2016年12月在宿州市立医院行单侧TKA的病人120例,利用抛硬币法随机分为A、B、C、D四组,每组各30例,均于止血带释放前关节囊缝合后局部关节腔注射药剂,其中A组注射50 mL生理盐水,B组注射含有10 mg/kg的TXA生理盐水50 mL,C组注射20 mg/kg的TXA生理盐水50 mL,D组注射含有30 mg/kg的TXA生理盐水50 mL,统计四组病人术后总引流量,术前、术后24 h和72 h的血红蛋白(hemoglobin,Hb),输血率以及术后并发症的发生率,分析四组间差异。结果在术后引流量方面,D组[(343.53±68.60) mL]＜C组[(404.57±62.59) mL]＜B组[(452.57±71.72) mL]＜A组[(532.67±87.05) mL],组间差异有统计学意义(F=35.847,P＜0.05);在Hb水平方面,四组术前差异无统计学意义(F=0.137,P=0.938),而在术后24 h和72 h方面,D组(120.70±9.60、110.83±11.04) mL＞C组(112.57±9.75、102.33±11.16) mL＞B组(105.50±9.84、93.30±11.95) mL＞A组(98.13±9.83、83.10±13.03) mL,组间差异有统计学意义(均P＜0.05);输血率方面,D组(3.33%)最低,A组(36.67%)最高,差异有统计学意义(χ²=11.779,P=0.008);并发症发生率方面,四组差异无统计学意义(χ²=3.025,P=0.388)。结论 关节腔注射TXA可以减少单侧TKA后病人的失血量和输血需求,不增加并发症的发生率,30 mg/kg为最佳剂量。     

医用耳脑胶局部注射治疗实质脏器外伤出血引起疼痛机制的动物实验研究

目的通过动物实验研究初步探讨超声引导下医用耳脑胶局部注射治疗腹部实质脏器外伤出血产生疼痛的可能机制。方法以腹腔注射医用耳脑胶及乙醇引起的小鼠扭体反应作为内脏痛的实验模型。NIH小鼠56只随机分为7组,每组8只动物,A组腹腔注射0.3mL生理盐水;B组分为B1、B2、B3亚组,分别腹腔注射医用耳脑胶0.1、0.2、0.3mL;C组分为C1、C2、C3亚组,分别腹腔注射75%乙醇0.1、0.2、0.3mL。观察各实验组30min的扭体反应,对疼痛程度进行计分。结果A、B组所有动物在30min内仅可见腹部的舔食行为,均未见明确的程度更重的扭体反应;疼痛程度计分A组为4.2±1.3,B1组为3.4±0.9,B2组为3.5±1.3,B3组为3.4±1.2;A组与B组各亚组间疼痛程度计分差异无统计学意义(P>0.05)。C组所有动物均可见不同程度的扭体反应,疼痛程度计分C1组为10.9±2.0,C2组为16.8±2.1,C3组为23.0±4.1,C组各亚组间及与A组、B组各亚组间疼痛计分差异有统计学意义(P<0.01)。结论医用耳脑胶局部注射治疗外伤出血引起疼痛主要机制可能为局部黏合的牵拉痛及局部的缺血性疼痛;医用耳脑胶可安全用于实质脏器超声引导下的微创止血治疗。     

谷氨酰胺和5—氨尿嘧啶对荷瘤大鼠肿瘤生长的影响

目的:探讨联合应用谷氨酰胺(Gln)和5-氟尿嘧啶(5-Fu)对肿瘤生长的影响。方法:制备荷Walker-256癌肉瘤大鼠模型,第6天随机分为A、B、C、D四组,分别腹腔注射生理盐水、丙氨酰谷氨酰胺二肽(Ala-Gln)、5-Fu、Ala-Gln和5-Fu。第14天检测瘤重、PCNA指数、凋亡指数、淋巴细胞转化率(LTR)及NK细胞活性(NKCA)。结果:治疗后B组瘤重(13.1±2.5)g与C组(12.0±1.2)g,无明显差异(P>0.05),B组PCNA、凋亡较C组无明显差异(P>0.05),B组LTR、NKCA较C组增强(P<0.05);治疗后D组(10.7±0.8)瘤重较B、C组明显降低(P<0.05),凋亡表达无明显差异(P>0.05),D组LTR较B、C组明显增高(P<0.05),NKCA较C组明显增高(P<0.05)。结论:联合应用Gln和5-Fu治疗肿瘤与单独应用Gln或5-Fu相比能够显著抑制肿瘤生长,且提高化疗后机体的免疫功能。     

辛伐他汀对骨质疏松大鼠血清骨钙素及骨密度的影响

目的:研究辛伐他汀对骨质疏松大鼠骨密度及血清骨钙素的影响,探讨其促进骨形成的作用。方法:36只雌性Wistar大鼠,随机分为4组。A组为假手术组,其余各组均行双侧卵巢切除术,术后1wk开始,A,B两组每只大鼠灌服生理盐水1.5mL·d-1,C组予辛伐他汀5mg·kg-1·d-1,D组予尼尔雌醇0.01mg·kg-1·d-1。10wk后处死大鼠,测量大鼠体重、股骨骨密度和血清骨钙素。结果:10wk后,B组大鼠体重高于A组,C,D组大鼠体重低于B组,均P<0.01。C,D组股骨骨密度(0.245±s0.007),(0.2430±0.0010)g·cm-2,高于B组(0.245±0.009)g·cm-2,血清骨钙素水平亦高于B组,均P<0.05,2组之间差异无显著意义(P>0.05)。结论:辛伐他汀能增加骨质疏松大鼠骨密度,提高骨钙素水平,促进新骨形成。     

曲马多复合加巴喷丁对糖尿病神经病理性疼痛大鼠脊髓胶质细胞活化的影响

目的观察曲马多复合加巴喷丁对糖尿病神经病理性疼痛大鼠脊髓胶质细胞活化及炎症因子表达的影响。方法健康雄性SD大鼠48只,6～7周龄,体重200～220 g,通过腹腔注射链脲佐菌素60 mg/kg建立糖尿病神经病理性疼痛模型。将造模成功大鼠随机分为A组(生理盐水组)、B组(曲马多20 mg/kg组)、C组(加巴喷丁60 mg/kg组)及D组(曲马多10 mg/kg+加巴喷丁30 mg/kg组),每组10只。A组:生理盐水10 ml/kg腹腔内注射,qd;B组:2 mg/ml曲马多溶液10 ml/kg腹腔注射,qd;C组:6 mg/ml加巴喷丁溶液10 ml/kg腹腔注射,qd;D组:1 mg/ml曲马多溶液+3 mg/ml加巴喷丁溶液10 ml/kg腹腔注射,qd。测定给药前1 d (T_0)及给药后3 d (T_1)、7 d (T_2)、11 d (T_3)时机械缩足阈值(PWT)和热缩足潜伏期(PWL),于第11天测定PWT和PWL后处死大鼠。采用免疫荧光双标法测定各组大鼠脊髓胶质细胞活化的情况,采用酶联免疫吸附法(ELISA)检测各组大鼠脊髓内肿瘤坏死因子α(TNF-α)和白细胞介素-1 (IL-1)的含量。结果 C组、D组机械缩足阈值高于A组、B组(P<0.05),T_3时D组机械缩足阈值高于C组,差异均有统计学意义(P<0.05)。与A组比较,B组T_2时热缩足潜伏期延长,C组T_1、T_2时热缩足潜伏期延长,D组T_1～T_3时热缩足潜伏期延长,差异均有统计学意义(P<0.05)。与B组、C组比较,D组T_3时热缩足潜伏期均延长(P<0.05)。与A组比较,B、C、D组大鼠脊髓内星状胶质细胞和小胶质细胞活化水平降低,TNF-α、IL-1表达降低,差异均有统计学意义(P<0.05)。与B组比较,C组和D组大鼠脊髓内星状胶质细胞和小胶质细胞活化水平降低(P<0.05)。与C组比较,D组大鼠脊髓内星状胶质细胞和小胶质细胞活化水平降低(P<0.05)。与B组、C组比较,D组大鼠脊髓TNF-α、IL-1表达降低,差异均有统计学意义(P<0.05)。结论曲马多复合加巴喷丁的镇痛效果优于单独应用曲马多或加巴喷丁,其机制可能与抑制胶质细胞的活化及减少TNF-α、IL-1表达有关。     

MSCs修饰受体胸腺对异种心脏移植免疫排斥反应的作用研究

目的大鼠胸腺以豚鼠骨髓间充质干细胞进行修饰,然后行豚鼠到大鼠的异种心脏移植。研究以间充质干细胞修饰胸腺对异种移植心脏存活时间的影响。方法供体豚鼠和受体SD大鼠各20只随机分成四组,A组(对照组);B组(胸腺修饰IT组);C组(CVF组);D组(IT加CVF组)。观测指标:移植心脏存活时间,病理变化,供受体异种淋巴细胞毒性试验。结果供心平均存活时间:A组(23.20±6.14)min,B组(24.20±9.28)min,C组(335.40±49.23)min,D组(451.00±50.10)min。A与B组两组比较,差异无统计学意义(P>0.05)。C、D组平均存活时间较A、B明显延长,差异有统计学意义(P<0.05)。D组与C组组间比较差异有统计学意义(P<0.05)。供受体淋巴细胞反应,测得光密度值分别为:D组为(381.20±12.01),A组为(590.20±15.38),胸腺修饰组明显低于对照组,差异有统计学意义(P<0.05)。结论豚鼠到大鼠心脏异种移植中,豚鼠MSCs修饰大鼠胸腺,当克服HAR后可以延长供心存活时间,但并不能完全克服AVR的发生。     

无细胞耻垢分枝杆菌疫苗对结核杆菌感染者的预防作用研究

目的初步探讨无细胞耻垢分枝杆菌疫苗对结核杆菌感染者的预防作用。方法1.动物实验:将结核杆菌感染的豚鼠随机分为不同剂量的无细胞耻垢分枝杆菌疫苗组(A组:7.0μg/次,B组:35.0μg/次)和阴性对照组(C组),观察疫苗对豚鼠PPD皮肤反应、病变指数和活菌负荷的影响。2.人体试验:将31名PPD皮试强阳性者随机分为不同剂量的无细胞耻垢分枝杆菌疫苗组(D组:8.7μg/次、E组:17.5μg/次、F组:35.0μg/次),观察受试者注射疫苗前后PPD皮肤反应情况。结果1.动物实验:A、B、C组豚鼠PPD皮肤反应强度依次为(9.87±3.36、7.04±3.09、12.67±1.47)mm,A组、B组和C组比较,均有极显著性差异(P<0.01);A、B、C组豚鼠病变指数依次为34.6±13.7、29.6±12.3、49.6±16.7,A组、B组和C组比较,均有显著性差异(P<0.05);A、B、C组豚鼠脾脏分离的结核杆菌对数值依次为4.32±1.58、4.00±1.39、4.94±0.41,B组和C组相比有显著性差异(P<0.05)。2.人体实验:D、E、F组受试者注射疫苗前PPD皮试结果依次为(36.1±4.5、34.0±3.6、33.5±3.1)mm,注射疫苗后PPD皮试结果依次为(20.6±0.6、16.0±1.6、17.82±1.2)mm,各组注射疫苗前后PPD皮试结果比较均有显著性差异(P<0.05)。结论无细胞耻垢分枝杆菌疫苗对结核杆菌感染者有较好的预防效果。     

七氟烷吸入麻醉对新生大鼠海马神经发生和认知功能发育的影响

目的探讨七氟烷吸入麻醉对新生大鼠海马神经发生和认知功能发育的影响。方法 7日龄SD大鼠64只,随机分为4组(n=16)。A组吸入40%氧气1 h,B、C、D组分别吸入1.0%、2.0%、4.0%七氟烷1 h。各组取8只大鼠,隔日予5-溴脱氧尿核苷(BrdU)50 mg.kg-1,ip,共3次。于14 d采用免疫组化评分(IHS)评估BrdU标记增殖细胞,观察4组海马神经发生情况。各组余8只大鼠于7 wk末采用Morris水迷宫测试其认知功能。结果 B、C、D组大鼠海马区BrdU阳性细胞IHS分别为2.74±0.40、2.30±0.20、2.30±0.31,A组为4.00±0.36,B、C、D组均低于A组(均P<0.05)。B、C、D组Morris水迷宫测试逃避潜伏期和探索时间均长于A组(P<0.05),且呈剂量依赖性。结论七氟烷吸入麻醉能抑制新生大鼠海马神经发生,并阻抑新生大鼠认知功能发育。     

A型肉毒毒素减轻面部整形切口瘢痕的临床观察

目的 观察A型肉毒毒素在减轻面部整形美容切口瘢痕中应用的临床疗效。方法 选取中国人民解放军空军军医大学第一附属医院2017年5—8月的面部整形手术病人32例,在手术结束关闭切口时将A型肉毒毒素随机注射到任意一半切口下为观察侧,另一半切口注射等量生理盐水为对照侧,比较病人两半切口愈合的瘢痕宽度、视觉模拟量表评分、温哥华瘢痕评分及病人满意度。结果 在瘢痕宽度［(0.31±0.12)mm比(0.45±0.17) mm,t＝2.252,P＝0.028］、视觉模拟量表评分［(6.10±3.82)分比(4.75±2.55)分,t＝2.834,P＝0.006］、温哥华瘢痕评分［(3.46±3.12)分比(6.62±3.47)分,t＝2.933,P＝0.005］及病人满意度［(9.21±1.32)分比(7.37±2.01)分,t＝2.120,P＝0.038］方面,注射A型肉毒毒素的切口侧优于对照侧,差异有统计学意义(P＜0.05)。结论 A型肉毒毒素可减轻面部整形美容术后切口瘢痕,产生更美观的术后外观,值得临床推广。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.