Poisonous snake bite is a common and critical disease in clinical practice.Untimely or improper treatment may lead to severe swelling, necrosis and even death of the patient’s limbs.This paper summarized the pathogenesis and treatment methods of local limb swelling and necrosis caused by venomous snake bite, analyzed the advantages and problems of traditional Chinese and western medicine in treating venomous snake bite, and selected the combination of traditional Chinese and western medicine for treatment of venomous snake bite.The standardization of the combination of traditional Chinese and western medicine in treating venomous snake bite needs to be further improved.

目的 采用小RNA(sRNA)测序技术检测清瘟败毒饮干预的脓毒症小鼠24?h脾组织微小RNA(miRNA)的表达变化,从而从非编码基因层面分析清瘟败毒饮抗脓毒性脾功能损伤的可能机制.方法 将45只SPF级健康BALB/C小鼠随机等分为对照组、脓毒症组和清瘟败毒饮组.清瘟败毒饮成分每剂含生石膏15?g、生地黄7.5?g、水牛角15?g、黄连2.5?g、栀子5?g、桔梗5?g、黄芩5?g、知母5?g、赤芍5?g、玄参5?g、连翘5?g、甘草5?g、丹皮5?g、竹叶5?g.脓毒症组和清瘟败毒饮组小鼠腹腔注射脂多糖(LPS)10?mg/kg构建脓毒症模型,对照组腹腔注射等量生理盐水.清瘟败毒饮组于制模前2?d开始给予清瘟败毒饮药液0.3?mL,每1?mL相当于生药1.5?g灌胃,术后继续胃饲,每日2次;对照组及脓毒症组给予等量生理盐水胃饲.术后24?h各组随机取5只小鼠脾组织,每组各取1只小鼠的脾组织于光镜下观察病理学改变,提取其余小鼠脾组织总RNA,采用sRNA测序技术进行miRNA检测,应用生物信息学软件分析脓毒症小鼠脾组织miRNA表达差异及靶基因的预测.?结果 光镜下显示,对照组小鼠脾组织结构正常;脓毒症组小鼠脾组织白髓总体结构稍紊乱,红白髓交界偏模糊,红髓充血明显;清瘟败毒饮组小鼠脾组织紊乱结构较前有所改善.与对照组比较,脓毒症组小鼠脾组织已知miRNA表达上调数为39个,下调数为16个;清瘟败毒饮组小鼠脾组织已知miRNA表达上调数为6个,下调数为24个.经LPS干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达明显上调;经清瘟败毒饮干预后,miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p表达出现一定程度下调.结论 清瘟败毒饮可改善脓毒症小鼠脾组织结构改变,可能通过下调miR-217-5p、miR-200b-5p、miR-429、miR-148A-3p基因表达起到保护脓毒症小鼠脾损伤的作用,可为将来进一步研究提供思路.

毒蛇咬伤是常见的急性中毒事件.毒蛇种类不同分泌的蛇毒液成分不同,可导致机体出现不同程度的局部组织坏死.血液毒和混合毒类毒蛇咬伤并发组织坏死事件的发生率较高,创面后期可能演变为感染、化脓、溃疡、坏死甚至肢体残障,这给患者带来许多后续的健康问题,为此学术界进行了大量的研究.本文就毒蛇咬伤致局部组织坏死的发病机制进行总结归纳,为临床诊疗提供参考及启发.

竹叶青蛇咬伤在我国南方地区的发病率较高,目前认为其咬伤机制大致为凝血功能障碍和炎症反应.近年来,我国传统医学治疗竹叶青蛇咬伤的方法与技术不断丰富,且具有确切的临床疗效,优势明显.本文就近年来中医治疗竹叶青蛇咬伤的研究进展作一概述,为竹叶青蛇咬伤的救治提供借鉴.

目的 研究蛇伤胶囊对竹叶青蛇伤血管内皮细胞Cyt-C/Bcl-2信号通路的影响.方法 20只新西兰大白兔被随机分为空白组和中药组,每组10只,分别制备常规兔血清和含药兔血清.人脐静脉血管内皮细胞(HUVEC)传代后随机分为正常组、模型组、低、中、高剂量含药血清组5组.正常组以10％正常兔血清培养,模型组在正常组的基础上以5μg/mL竹叶青蛇毒液培养,低、中、高剂量组分别在模型组基础上培养6 h后加入5％、10％、15％的含药兔血清培养.各组培养72 h后,收集细胞,流式细胞术检测HUVEC细胞凋亡率,Western blot法检测HUVEC细胞浆及线粒体内Cyt-C、AIF蛋白表达,RT-qPCR法检测HUVEC中Bax、Bak、Bad、Bid、Bcl-xl、Bcl-w、Bcl-2、Caspase-3、Caspase-9 mRNA表达.结果 与正常组比较,模型组细胞凋亡率明显提高(P<0.01);细胞浆Cyt-C、AIF的蛋白表达明显提高(P<0.01),线粒体内Cyt-C、AIF的蛋白表达明显下降(P<0.01);Bax、Bak、Bad、Bid、Caspase-3、Caspase-9 mRNA表达明显提高(P<0.01),Bcl-xl、Bcl-w、Bcl-2 mRNA表达明显下降(P<0.01).与模型组比较,低、中、高剂量组细胞凋亡率明显降低(P<0.01);细胞浆Cyt-C、AIF蛋白表达明显降低(P<0.01),线粒体内Cyt-C、AIF蛋白表达明显提高(P<0.01);Bax、Bak、Bad、Bid、Caspase-3、Caspase-9 mRNA表达明显降低(P<0.01),Bcl-xl、Bcl-w、Bcl-2 mRNA表达明显提高(P<0.01).与中剂量组比较,低、高剂量组细胞凋亡率明显增加(P<0.01);细胞浆Cyt-C、AIF蛋白表达明显提高(P<0.01),线粒体内Cyt-C、AIF蛋白表达明显下降(P<0.01);Bax、Bak、Bad、Bid、Caspase-3、Caspase-9 mRNA表达明显增加(P<0.01),Bcl-xl、Bcl-w、Bcl-2 mRNA表达明显下降(P<0.01).结论 蛇伤胶囊能够通过调控Cyt-C/Bcl-2信号通路抑制竹叶青蛇伤血管内皮细胞凋亡,从而治疗竹叶青蛇伤及其引起的血管损伤.

目的 采用Small RNA测序技术检测脓毒症小鼠24 h脾组织microRNA(miRNA)的表达变化,并从非编码基因层面分析脓毒症脾功能损伤的可能机制.方法 将30只SPF级健康BALB/C小鼠随机分为对照组和脓毒症组,每组15只.脓毒症组采用LPS 10 mg/kg腹腔注射进行建模,对照组仅予等量生理盐水腹腔注射.两组术毕均行肌肉注射平衡液5 ml/kg,并于术后24 h各组随机取5只小鼠的脾组织提取RNA,采用Small RNA测序技术进行miRNA检测,然后应用生物信息学软件分析脓毒症小鼠脾组织的miRNA表达差异及靶基因预测.结果 小鼠建模后24 h,与对照组相比,脓毒症小鼠脾脏组织已知及未知miRNA表达上调数分别为39个、3个,下调数分别为16个、3个.其中已知miRNA表达上、下调倍数最大的前5个基因分别为Rno-miR-217-5p,rno-mir-216a-5p,rnomiR-216b-5p,rno-miR-375-3p,rno-mir-216b-3p.下调倍数前5的miRNA:Rno-miR-138-5p,rno-miR-451a-3p,rno-miR-138-1-3p,rno-miR-202-5p,rno-miR-592.结论 LPS诱导脓毒症小鼠可出现miRNA表达量的改变,其上、下调倍数最显著的miRNA可能对预测LPS诱导的脓毒症脾功能损伤有一定指导意义.

目的 对比脓毒症组小鼠与正常对照组小鼠的肠道菌群,以明确肠道菌群组成特点.方法 将60只BALB/C小鼠随机等分为对照组(CF组)和脓毒症组(SF组)(每组各30只).SF组小鼠按脂多糖(LPS)10 mg/kg腹腔注射构建脓毒症模型;CF组则予等量生理盐水腹腔注射.术后24、72 h每组小鼠留取粪便标本行16sRNA测序.在72 h同时取盲肠组织做组织学检查.结果 LPS干预后72 h小鼠结肠局部黏膜上皮缺失,肠腺结构消失,炎性细胞浸润,上皮细胞肿胀.肠道菌群方面:SF组较CF组小鼠的肠道菌群多样性明显减少.CF组小鼠肠道菌群主要以拟杆菌门和厚壁菌门为主,但SF组小鼠厚壁菌属比例总体呈现减少趋势,变形菌门则表现为增长的趋势.LPS干预后24 h SF组小鼠肠道菌群出现紊乱:粪肠球菌属、大肠杆菌属、弗格森埃希菌属、缓慢葡萄球菌属及不可培养的大肠杆菌属明显增多(P<0.05),梭状芽孢菌属、毛螺科菌属、未培养的梭菌属则明显减少(P<0.05).LPS干预后72 h SF组小鼠肠道菌群的变化情况:铅黄肠球菌、粪肠球菌、大肠杆菌、费格森埃希菌、金氏拟杆菌、奇异变形杆菌、未培养的埃希菌属和肠球菌明显增多(P<0.05),鼠乳杆菌、未培养的拟杆菌属、约氏乳杆菌则明显减少(P<0.05).结论 脓毒症小鼠可致肠道微生态的紊乱,微生物的多样性减少,潜在致病菌的过度生长,同时炎症作用下肠道屏障的破坏可能为微生物移位创造条件,也可能成为病情进一步发展的重要推动力.

目的 采用16sRNA测序技术检测大黄对脓毒症小鼠早期肠道菌群的变化情况,分析大黄对脓毒症小鼠的治疗作用机制.方法 将45只BALB/C小鼠随机等分为3组,即对照组、脓毒症组、脓毒症大黄干预组.实验前正常饮食、饮水.术前2 d脓毒症大黄干预组按生药量50 mg/kg灌胃给药,每天2次;对照组、脓毒症组给予等量生理盐水灌胃.3组小鼠灌胃给药2 d后,脓毒症组及脓毒症大黄干预组小鼠按LPS 10 mg/kg腹腔注射构建脓毒症模型,对照组予等量生理盐水腹腔注射.建模后24 h取盲肠组织行病理检查以及粪便组织16sRNA检测.结果 经大黄干预后,脓毒症小鼠盲肠组织结构的改变得到一定程度的缓解.脓毒症组小鼠厚壁菌属比例明显减少,拟杆菌属相对增多.脓毒症大黄干预组小鼠拟杆菌属、厚壁菌属、弯曲菌属均有减少,而变形菌属明显增多.结论 大黄干预后,脓毒症小鼠肠道菌群进一步变化,变形杆菌属相对增多为优势菌群;结合病理结果,考虑大黄对脓毒症小鼠的治疗作用主要是通过保护肠道黏膜屏障而起到保护作用.

目的:采用RNA-seq技术检测脓毒症大鼠24h、48h脾组织mRNA的表达变化，从而分析脓毒症脾功能障碍可能的机制。方法:运用随机数字表法将30只清洁级健康雄性Wistar大鼠等分为对照组、脓毒症组。脓毒症组采用盲肠结扎穿孔术(CLP)来建模。对照组仅行开腹、关腹。2组术毕均肌肉注射平衡液5 mL/kg。术后24 h、48 h各组随机取5只大鼠取脾组织提取RNA后采用RNA-seq进行mRNA检测，应用生物信息学软件分析脓毒症大鼠脾组织mRNA随时间变化的表达差异及涉及的相关通路。结果:大鼠建模后24、48小时，相对于对照组，脓毒症大鼠脾脏组织mRNA表达上调数分别为1 030个，1 354个，下调数分别为935个，1 763个。其中，脓毒症大鼠脾组织随时间变化，细胞因子-细胞因子受体相互作用通路及凋亡相关通路相关mRNA表达差异明显。脓毒症大鼠脾组织细胞因子及其受体相互作用相关信号通路的mRNA表达情况为脓毒症24 h大鼠脾组织mRNA表达上调，而48 h转为下调的基因有2个，上调转为正常表达有22个，正常表达转为下调有30个，表达下调转为上调有1个，下调转为正常表达有2个，正常表达转为上调有10个。细胞凋亡相关通路中，脓毒症大鼠24 h脾组织mRNA表达上调，而48 h转为表达下调有1个，从表达上调到正常表达有5个，从正常表达到表达下调有9个，从正常表达到表达上调有10个，从表达下调到表达上调有1个，从表达下调转为正常表达有1个。结论:早期过度炎症反应，晚期免疫抑制是脓毒症重要机制之一。其机制可能跟细胞-细胞因子受体、凋亡通路相关基因表达有关。

目的 观察犀角地黄汤对脓毒症大鼠肝组织mRNA表达的影响.方法 将45只Wistar大鼠按随机数字表法分为假手术组、脓毒症模型组和犀角地黄汤干预组,每组15只.采用盲肠结扎穿孔术(CLP)复制脓毒症大鼠模型;假手术组按同法手术,但不行CLP.各组术后肌肉注射平衡液5 mL/kg.犀角地黄汤组于术前2 d开始灌胃犀角地黄汤(组成:水牛角30 g、生地黄24 g、赤芍12 g、牡丹皮9 g),相当于生药3 g,每日2次,术后再连续灌胃2 d.2 d后各组随机选择5只大鼠取肝组织提取总RNA,采用RNA-seq检测肝组织mRNA;光镜下观察各组肝组织病理学改变;应用生物信息学软件分析犀角地黄汤干预后大鼠肝组织mRNA差异表达及所涉及的相关通路.结果 光镜下可见:犀角地黄汤干预组大鼠肝汇管区炎性细胞浸润较脓毒症模型组明显减轻.犀角地黄汤干预组大鼠肝组织mRNA上下调数较脓毒症模型组明显增多,脓毒症模型组大鼠肝组织mRNA上调914个、下调822个,犀角地黄汤组肝组织mRNA上调2263个、下调1253个(均P<0.05).犀角地黄汤干预组肝组织辅助性T细胞1(Th1)和Th2细胞分化通路相关mRNA表达下调,而犀角地黄汤干预组恢复正常表达的有9个,表达上调转正常表达4个,正常表达转为表达上调5个,表达下调转表达上调有1个,正常表达转为表达下调有1个,同时上调的有4个.B细胞受体通路相关差异mRNA表达上调恢复正常表达的有14个,正常表达转表达上调4个,正常表达转表达下调有1个,同时上调的3个.Toll样受体(TLR)信号通路相关差异mRNA表达上调恢复正常表达的8个,正常表达转表达上调的7个,表达下调转正常表达的5个,正常表达转表达下调的2个,同时上调的10个,同时下调的3个.结论 犀角地黄汤可改善脓毒症大鼠肝组织病理学改变,其机制可能与犀角地黄汤调节Th1和Th2分化、B细胞受体通路及TLR信号通路相关mRNA的表达有关.

我国地域辽阔,毒蛇种类丰富.依托蛋白组学分析,对毒素成分有了相对完整的认识,其中眼镜蛇毒素的临床应用是研究热点.在不同给药途径时,其毒素均能产生镇痛作用,可能与胆碱能系统、内源性阿片系统或补体旁路系统有关;在抑制炎症方面可以提高抗炎因子、降低致炎因子达到平衡状态,起到抑炎作用;在抗肿瘤方面认为可以促进癌细胞的自噬、凋亡;在免疫抑制方面认为可以抑制T细胞周期.本文总结了眼镜蛇毒素各方面临床应用作用及其可能机制,为进一步去挖掘提供方向.

Objective:To investigate the influence of purging fire and removing toxinon therapy on the related factors of vascular endothelial cells nuclear transcription factor kappa B (NF-kB) signaling pathway in Trimeresurus stejnegeri Bites.Methods:The animal and cell models were established according to the former study.The blank group (KB),the model group (MX) and the low,middle and high dose Sheshang capsule liquid groups (DY,ZY,GY) were set up.There were 10 samples in each group.In vivo,KB was injected 0.75 mL/kg normal saline into rabbits' right hind legs,and received 10 mL/kg normal saline by gavage after 6 hours.MX,DY,ZY and GY were injected 0.75 mL/kg Trimeresurus Stejnegeri venom into rabbits' right hind legs.After 6 hours,MX received 10 mL/kg normal saline by gavage,and Sheshang Capsule groups with different doses received 10 mL/kg low,middle and high dose Sheshang Capsule liquid by gavage respectively.The gavage was given once a day for a week.24 hours after the last gavage,the blood of the rabbits was collected through auricular vein and the serum was separated.In vitro,KB was cultured with 10％ blank rabbit serum.MX was cultured with 5 μg/mL of snake venom in addition.DY,ZY and GY were cultured with the same composition as MX in the first 6 hours and then they were cultured with 5％,10％ and 15％ drug serum respectively in addition.After 72 hours culture,the human vascular endothelial cells(HUVEC) and medium were collected to measure.The level of IkBs kinase (IKK),nuclear transcription factor kappa B (NF-kB) and NF-kB induced kinase (NIK) in rabbit serum were measured by enzyme linked immunosorbent assay (ELISA).Protein expressions of IKK,NIK and NF-kB p65 in vascular endothelial cells of each group wre detected by Western blot.T he expression of NF-kB p65 in vascular endothelial cells was detected by confocal laser scanning microscope (LSCM).Results:In animal experiment,the levels of IKK,NIK and NF-kB p65 in MX group increased significantly compared with those of KB group (P ＜ 0.01).In ZY group and GY group,the levels of IKK,NIK and NF-kB p65 all decreased compared with those of MX group (P ＜ 0.05 or P ＜ 0.01).In vitro cell experiment,the protein expressions of IKK,NIK and NF-kB p65 in vascular endothelial ceils in MX group increased significantly compared with those of KB group (P ＜ 0.05 or P ＜ 0.01).In DY group and ZY group,the protein expressions of IKK,NIK and NF-kB p65 all decreased compared with those of MX group (P ＜ 0.05or P ＜0.01).The expression of NF-kB p65 in HUVEC in MX group was significantly higher than that in KB group under LSCM (P ＜ 0.01).In DY group and ZY group,the expressions of NF-kB p65 in HUVEC were decreased compared with those of the MX group(P ＜ 0.01).Conclusion:NF-kB signaling pathway is the key to inflammatory injury induced by Trimeresurus stejnegeri bites in vascular endothelial cell.The therapy of purging fire and removing toxin could treat inflammatory injury of the vascular endothelial cell induced by Trimeresums stejnegeri bites through the regulation of NF-kB signaling pathway.

Objective To investigate the influence of the purging fire and removing toxin method on chemokines and adhesion factors related to vascular endothelialitis injury induced by toxin of trimeresurus stejnegeri bite.Methods ① Animal experiment:50 healthy New Zealand white rabbits were chosen.According to random numbers generated by statistical software,they were divided into normal control group,model group,low,middle and high dose Sheshang capsule groups,10 in each group.Trimeresurus stejnegeri bite model was replicated by injecting 0.75 mL/kg snake venom into subcutaneous tissues of rabbits' right hind legs.And the same volume of normal saline was injected into the rabbit in the normal control group.After the model was established for 6 hours,the rabbits in low,middle and high dose Sheshang capsule groups received 174,348 and 522 mg· kg-1 · d-1 of Sheshang capsule solution respectively (the content of capsules was dissolved in normal saline to make liquid with 17.4,34.8 and 52.2 g/L Sheshang solution respectively,so the volume of gavage of each group was 10 mL· kg-1 · d-1);in the model and normal control groups,the same amount of normal saline was given by gavage,once daily for consecutive one week.24 hours after the last gavage,the blood of the rabbits was collected through an auricular border vein and the serum was separated by centrifuge ready for use.Meanwhile,the whole abdominal aorta segment of the rabbit was harvested and kept them in liquid nitrogen ready for use.② Cell experiment:human umbilical vascular endothelial cell (HUVEC) was cultured with MEM for 24 hours.The solution was replaced and according to the random number generated by statistical software,the cells were divided into blank control group,model group and low,middle,high dose Sheshang capsule medicinal serum groups,10 wells in each group.Trimeresurus stejnegeri toxin cell model was reproduced by addition of 5 mg/L snake venom into the cell culture medium.After 6-hour culture,the cells of model group and blank control group received 10％ normal rabbit serum,and the cells of low,middle and high dose Sheshang medicinal serum capsule groups received serum containing 5％,10％ and 15％ drug,respectively.After culture for 72 hours,the cells were collected and the total RNA was extracted.The real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to detect the levels of mRNA of interleukin-8 (IL-8),monocyte chemoattractant protein-1 (MCP-1),intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial cell adhesion molecule-1 (VCAM-1) in the vascular endothelial cells of rabbit aorta abdominalis and human umbilical vein,and the content of serum E-select element (CD62E) was measured by enzyme linked immunosorbent assay (ELISA).Results In model group,the expression levels of mRNA in IL-8,MCP-1,ICAM-1,VCAM-1 and the content of CD62E were all increased significantly in the endothelial cells of rabbit aorta abdominalis and HUVEC compared with those in control group [when the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 in normal and blank control group were all being 1,the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in animal model group were 3.96 ± 0.39,3.07 ± 0.27,3.71 ± 0.26,3.94 ± 0.26,and the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in HUVEC model group were 3.53±0.70,2.24±0.48,3.13±0.44,2.80±0.13,respectively,all P ＜ 0.01].The content of CD62E in serum was increased significantly in model group compared with that in normal control group (μg/L:1.31 ± 0.22 vs.0.82 ± 0.13,P ＜ 0.01),the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 were decreased significantly in low,middle,high dose Sheshang capsule groups compared with those in model group in endothelial cells of aorta abdominalis of rabbits and HUVEC [abdominal aorta:IL-8 mRNA (2-△ △Ct) were 1.13 ± 0.19,1.26 ± 0.16,1.27 ± 0.17 vs.3.96 ± 0.39,MCP-1 mRNA (2-△ △ Ct) were 1.79 ± 0.24,2.22 ± 0.38,1.76±0.19 vs.3.07±0.27,ICAM-1 mRNA (2 △△Ct) were 2.05±0.11,1.68±0.09,2.37±0.48 vs.3.71±0.26,VCAM-1 mRNA (2-△△Ct) were 1.59±0.08,1.40±0.11,1.84±0.11 vs.3.94±0.26;HUVEC:IL-8 mRNA (2-△△Ct) were 2.33±0.59,2.82±0.82,2.51±0.77 vs.3.53±0.70,MCP-1 mRNA (2-△△Ct) were 1.59±0.35,1.48±0.36,1.54±0.29 vs.2.24±0.48,ICAM-1 mRNA (2-△△Ct) were 1.46±0.38,1.77±0.65,1.73±0.50 vs.3.13±0.44,VCAM-1 mRNA (2-△△Ct) were 2.49±0.24,2.18±0.19,2.45±0.24 vs.2.80±0.13,all P ＜ 0.05].The contents of CD62E were decreased significantly in middle,high dose Sheshang capsule groups compared with the content in model group (μg/L:1.01 ±0.14,1.04±0.13 vs.1.31 ±0.22,all P ＜ 0.01),but there were no statistical significant differences among the three drug group (all P ＞ 0.05).Conclusion The therapy of purging fire and removing toxin can treat vascular endothelial injury by inhibiting the inflammatory response induced by Trimeresurus stejnegeri bites.

目的 探讨加减荆防败毒散对风寒型外感热病的疗效及其对患者血清肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)的影响. 方法 将100例风寒型外感热病患者按随机数字表法分为治疗组和对照组,对照组予西药治疗,治疗组应用加减荆防败毒散治疗,观察2组患者退热时间、中医证候积分改变及疗效;选取健康志愿者30例作为健康组,测量治疗组和对照组治疗前后及健康组血清TNF-α、IL-1β含量. 结果 治疗组退热时间短于对照组,中医证候改善及疗效优于对照组(P＜0.05),2组患者治疗前血清TNF-α、IL-1β含量均较健康组明显升高(P＜0.05),治疗后血清TNF-α、IL-1β含量均明显下降(P＜0.05),治疗组优于对照组(P＜0.05). 结论 加减荆防败毒散可能影响外感热病(风寒证)患者血清TNF-α、IL-1β水平而发挥抗炎作用,提高临床疗效.

目的:观察竹叶青蛇伤患者中医辨证饮食护理的临床效果.方法:收集竹叶青蛇咬伤患者120例,随机分为观察组和对照组各60例,对照组给予一般饮食指导,观察组给予中医辨证膳食指导.观察2组疗效,平均住院天数、住院金额及患者满意度情况.结果:痊愈率观察组为71.7％,对照组为50.0％,组间比较,差异有统计学意义(P＜0.05).2组住院天数、住院金额、满意度比较,差异均有统计学意义(P＜0.05).结论:应用中医辨证饮食护理竹叶青蛇伤患者临床效果满意.

目的 观察蛇伤胶囊对竹叶青蛇伤血管内皮细胞IL-8、VCAM-1、MIP-1α表达的影响.方法 根据课题组前期实验方法制备蛇伤胶囊兔含药血清及复制竹叶青蛇伤细胞模型,设空白组(K B组)、模型组(M X组)和低、中、高浓度蛇伤胶囊药液治疗组(DY组、ZY组、GY组)5组,每组10个样本.在细胞实验中,KB组加入10%正常兔血清培养,MX组在KB组的基础上加入5μg/ml竹叶青蛇毒液培养,DY组、ZY组和GY组在MX组基础上培养6 h后分别加入5%、10%和15%的含中药兔血清继续培养.各组分别培养72 h后,收集细胞培养液,以酶联免疫吸附法检测细胞培养液中的IL-8、VCAM-1、MIP-1α含量.结果 MX组的IL-8、VCAM-1及MIP-1α水平相对KB组均显著升高(均P<0.01),蛇伤胶囊药液治疗组IL-8、VCAM-1及MIP-1α水平相对MX组均不同程度降低,其中ZY组变化有统计学意义(P<0.01或P<0.05).结论 蛇伤胶囊可通过降低 IL-8、VCAM-1及 MIP-1α表达治疗竹叶青蛇伤所致血管内皮炎症反应.

目的 研究黄葛香草汤治疗湿热型急性细菌感染性腹泻的临床疗效. 方法 120例湿热型急性细菌感染性腹泻患者按电脑随机数分成对照组、观察组各60例.对照组口服左氧氟沙星片、补液盐;观察组在对照组治疗的基础上,加服黄葛香草汤;2组均治疗3天.观察两组的临床疗效,检测2组患者治疗前后白细胞(WBC)、中性粒细胞(NE)、超敏C-反应蛋白(hs-CRP)和降钙素原(PCT)水平. 结果 观察组疗效优于对照组(P＜0.05).治疗后2组WBC、NE、hs-CRP、PCT水平均较治疗前降低(P＜0.01),且观察组优于对照组(P＜0.05或P＜0.01). 结论 黄葛香草汤治疗湿热型急性细菌感染性腹泻的临床疗效优于单纯西药抗菌治疗,具有抗肠道细菌感染的作用.

世界范围内眼镜蛇属约28种［1］。我国有舟山眼镜蛇（naj a atra）和孟加拉眼镜蛇（naj a kaouthia）两种，前者过去又称为中华眼镜蛇［2］。眼镜蛇是腺管牙类毒蛇，其毒腺分泌的毒液是以神经毒为主的混合毒，即神经毒和血循毒。眼镜蛇毒神经毒素（cobra neurotoxin ，CNT）是其毒液中毒性最大的成分，它能使动物产生呼吸抑制和骨骼肌麻痹，甚至死亡［3］。随着分子生物学和蛋白质组学的发展，眼镜蛇毒神经毒素的一些组分已得到分离、纯化、测定，并广泛应用于科研和临床。本文就眼镜蛇毒组分中神经毒素的研究与应用进展综述如下。

目的 研究泻火解毒法对竹叶青蛇伤血管内皮细胞炎性因子的影响.方法 ①动物实验:选择50只健康新西兰大白兔,根据计算机统计软件产生的随机数分为5组,每组10只.采用经兔右后腿皮下注射0.75 mL/kg竹叶青蛇毒液的方法复制竹叶青蛇伤动物模型;对照组经兔右后腿皮下注射等量生理盐水.制模后6h,低、中、高剂量蛇伤胶囊组给予蛇伤胶囊,剂量分别为174、348和522 mg· kg-1·d-1,使用时以生理盐水稀释为17.4、34.8和52.2 g/L的药液,灌胃量则均为10 mL· kg-1·d-1;对照组和模型组灌胃等量生理盐水;各组均每日灌胃1次,连续1周.于末次灌胃后24 h经耳缘静脉取血,分离血清,用五分类血细胞计数仪检测血中白细胞计数(WBC)和单核细胞(MO)数;用酶联免疫吸附试验(ELISA)测定血清白细胞介素(IL-1、IL-2、IL-6)和肿瘤坏死因子-α(TNF-α)水平.②细胞实验:用MEM培养液培养人脐静脉内皮细胞(HUVEC) 24 h后更换培养液并按随机数字表法分为5组,每组10个样本.采用细胞中加入5 mg/L竹叶青蛇毒液的方法复制竹叶青蛇毒细胞模型.培养6h后,空白对照组和模型组给予10％正常兔血清培养液,低、中、高剂量蛇伤胶囊组分别加5％、10％和15％含中药兔血清继续培养.培养72 h后,收集细胞培养液,用ELISA试验检测HUVEC培养液中IL-1、IL-2、IL-6、TNF-α水平.结果 模型组血WBC、MO、IL-1、IL-2、IL-6、TNF-α和HUVEC培养液中IL-1、IL-2、IL-6、TNF-α水平均较对照组明显升高,随着药物剂量增加,低、中、高剂量蛇伤胶囊组上述指标均较模型组下降,以中剂量蛇伤胶囊组降低更显著[血清:WBC(×109/L)为9.03±2.16比12.11±4.38,MO(×109/L)为0.44±0.18比0.63±0.16,IL-1 (ng/L)为92.46±9.46比110.80±29.78,IL-2(ng/L)为92.11±17.55比112.0±18.83,IL-6 (ng/L)为481.73±147.12比667.26±226.41,TNF-α(ng/L)为7.12±2.96比9.41±1.76;HUVEC培养液:IL-1 (ng/L)为56.76±10.37比75.80±22.49,IL-2 (ng/L)为76.77±13.73比92.80±17.82,IL-6(ng/L)为231.70±107.91比413.25±178.65,TNF-α(ng/L)为223.98±30.31比252.86±30.75,均P＜ 0.05].结论 竹叶青蛇毒可造成血管内皮细胞炎性损伤;泻火解毒法可治疗竹叶青蛇伤血管内皮炎性损伤.

目的 研究蛇伤胶囊治疗竹叶青蛇伤凝血障碍的机制.方法 根据前期实验复制竹叶青蛇伤的动物及细胞模型,均设置空白组(KB),模型组(MX),低、中、高剂量蛇伤胶囊药液组(DY、ZY、GY),共5组,每组10个样本.在动物实验中,KB组经兔右后腿皮下注射0.75 ml/kg生理盐水,6 h后灌胃10 ml/kg生理盐水.MX、DY、ZY和GY组经兔右后腿皮下注射0.75 ml/kg竹叶青蛇毒液,6 h后分别灌胃10 ml/kg生理盐水和10 ml/kg低、中、高剂量蛇伤胶囊药液.各组均每日灌胃1次,连续灌胃1周.于末次灌胃后24 h经耳缘静脉采血,分离血清.在细胞实验中,KB组加入10%正常兔血清培养,MX组在KB组的基础上加入5μg/ml竹叶青蛇毒液培养,DY、ZY、GY组在MX组基础上培养6 h后分别加入5%、10%、15%的含中药兔血清继续培养,各组分别培养72 h后,收集细胞培养液.以酶联免疫吸附法检测兔血清及细胞培养液中组织型纤溶酶原激活剂(tissue plas-minogen activator,t-PA)、纤溶酶原激活物抑制物-1(plasminogen activator inhibitor-1,PAI-1)和抗凝血酶-Ⅲ(Antithrombin-Ⅲ,AT-Ⅲ)水平.结果 MX组的AT-Ⅲ和t-PA水平相对KB组均显著升高(P<0.01),PAI-1水平较KB组显著降低(细胞实验中,P<0.05;动物实验中,P<0.01);DY、ZY和GY组的AT-Ⅲ 及t-PA水平相对MX组均明显降低(动物实验:ZY组t-PA和DY组AT-Ⅲ,均P<0.05;细胞实验:ZY组AT-Ⅲ,P<0.05;其余均P<0.01),DY、ZY和GY组的PAI-1水平较MX组明显升高(P<0.01).结论 蛇伤胶囊具有促凝和抗纤溶、改善机体凝血功能的作用,是蛇伤胶囊治疗竹叶青蛇伤凝血障碍的部分机制.