G—CSF和IL—6联合应用协同促进受照小鼠造血功能恢？…

目的 探讨Ｇ－ＣＳＦ和ＩＬ－６联合应用协同促进受辐射小鼠血血功能恢复的作用及机理。方法 采用Ｆ８００血细胞自动计数仪计数红细胞、白细胞、血小板和血红蛋白等指标。常规法进行骨髓集落培养,内生脾结节计数,琼脂糖电泳方法检测骨髓细胞程序性死亡。结果 Ｇ－ＣＳＦ和ＩＬ－６联合应用促进受辐射小鼠外周血细胞的恢复,促进脾脏内源性脾结节的形成,增加骨髓细胞含量和促进骨细胞ＧＭ－ＣＳＦ的形成,提高受辐射小鼠的生存     

艰难梭菌毒素AC端结构域在大肠杆菌中的可溶性表达与卵黄抗体的制备

目的重组表达艰难梭菌毒素A的C端结构域,制备针对艰难梭菌毒素A C端结构域的卵黄抗体。方法将优化合成的艰难梭菌毒素A的C端结构域DNA片段克隆到表达载体pET32b（＋）上,重组质粒转入大肠杆菌中诱导表达,经高亲和镍离子树脂纯化得到融合蛋白,凝血酶酶切后用高亲和镍离子树脂吸附多余多肽获得目的蛋白。检测目的蛋白的生物学活性,用目的蛋白免疫产蛋鸡制备针对艰难梭菌毒素A受体结合区的鸡卵黄抗体,分离纯化鸡卵黄抗体并用ELISA测定卵黄抗体的效价。结果获得了优化的艰难梭菌毒素A受体结合区的基因片段,继而制备了具有生物学活性的毒素A C端结构域重组蛋白。免疫产蛋鸡后获得了效价1∶50 000的抗艰难梭菌毒素A的卵黄抗体。结论获得了具有生物学活性的艰难梭菌毒素A C端结构域重组蛋白,为建立艰难梭菌基因工程疫苗打下了基础,并制备了针对艰难梭菌毒素蛋白A的卵黄抗体,可用于艰难梭菌相关性腹泻的诊断和治疗。     

中国大陆庚型肝炎病毒结构区与非结构区部分cDNA基因的序列分析

作者在建立检测庚型肝炎病毒酶免疫和ＰＣＲ技术的基础上，将抗－ＨＣＶ上阳性病人血清经逆转录－巢式ＰＣＲ分离部分ＨＧＶｃＤＮＡ片段，用与ＨＧＶ基因互补的特异寡核苷酸引物进行双脱氧核苷酸链末端终止法－ＰＣＲ直接测序。结果表明，中国大陆ＨＧ－３０２Ⅰ株部分ｃＤＮＡ序列与Ｌｉｎｎｅｎ等报道的ＨＧＢ美国株ｃＤＮＡ序列有极高的同源性。     

免疫透射和免疫散射比浊法测定血清免疫球蛋白及补体的比较

目的：比较免疫透射和免疫散射比浊法测定免疫球蛋白和补体时的准确性、精密度、偏离度及测定范围。方法：采用贝克曼LX-20与DELTA特定蛋白分析仪，分别以免疫透射和免疫散射比浊法，对同一临床标本测定血清中免疫球蛋白（IgG、IgA、IgM）和补体（C3、C4）的含量。结果：两种仪器测定IgG、IgA、IgM、C3、CA准确性、精密度均良好。两法各浓度值测定结果的预期偏差均在可接受范围。结论：贝克曼LX-20应用免疫透射比浊法与DELTA特定蛋白分析仪应用免疫散射比浊法测定免疫球蛋白和补体准确性、精密度均比较好，且散射比浊法优于透射比浊法。     

Four nonfatal and six fatal cases of opiate use: utility of morphine, its metabolites, and their ratios in blood specimens

Four nonfatal and six fatal cases of opiate use are presented with careful toxicological analysis. Levels of morphine (M), 6-monoacetylmorphine (6-MAM), morphine-6-glucuronide (M6G), and morphine-3-glucuronide (M3G) in blood specimens were measured by the sophisticated method of liquid chromatography (LC)-electrospray ionization (ESI)-tandem mass spectrometry (MS-MS). Fatal cases were characterized by much higher levels of free M than the nonfatal cases; this caused lower ratios of M6G/M and M3G/M in the fatal cases when compared with the nonfatal cases. Among the six fatal cases, the M6G/M ratios were especially low in four cases, in which rapid deaths were estimated. The present data are compared with data previously reported by other groups, and we discuss the utility of the levels of M, M6G, and M3G in blood and their ratios for estimating the antemortem status of each individual.     

Role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation and the effect of bifidobacterial supplement on gut barrier function following burns

Early multiple organ dysfunction syndrome appears to be facilitated with bacterial translocation in severely burn injury, yet the mechanisms of bacterial translocation remains in dispute. The aim of this study was to investigate the potential role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation following burns and the effects of bifidobacterial supplement on gut barrier. Methods: Wistar rats were randomly divided into burn group (Burn, n=60),sham burn group (SB, n=10) in experiment Ⅰ , and burn + saline group (BS, n=30), burn + bifidobacteria group (BB, n=30), and sham-burn + saline group (SS, n= 10) in experiment Ⅱ. Animals in BB group were fed bifidobacterial preparation (5 × 109 CFU/ml) after burns, 1.5ml,twice daily. Animals in BS and SS were fed saline. Samples were taken on days 1, 3, and 5 in burn groups, and on day 3 in sham-burn groups. The incidence of bacteria/endotoxin translocation and counts of Bifidobacterium, Fungi and Escherichia coli in gut mucosa were determined with standard methods. The levels of sIgA in mucus of small intestine were measured by RIA. The positive sIgA expression in lamina propria and ileum mucosal injury was evaluated light microscopically by blinded examiners. Results: Our results showed that the incidence of bacterial translocation was increased after burns, which was accompanied by significant decrease in number of bifidobacteria but significant increase in E. coli and fungi in gut mucosa, and elevation of levels of plasma endotoxin and IL-6 (P<0. 001).The incidence of bacterial translocation was markedly reduced after 3- and 5-day supplementation of bifidobacteria compared with control group (P<0.05). The counts of mucosal bifidobacteria were increased by 4- to 40-fold,while E. coli and fungi were decreased by 2- to 30-fold and 10- to 150-fold, respectively, after bifidobacterial supplementation in contrast to control group. The damage of mucosa tended to be less pronounced after 3-day bifidobacteria-supplemented formula compared with control group [grade 2(0-6) vs. grade 4(3-6), P<0.05]. Moreover, the expression and release of sIgA was markedly augmented after 3-day bifidobacteria-supplementation formula and it returned to normal range on day 5. Conclusion: The decrease in counts and proportion of bifidobacteria in mucous membrane flora may play an important role in the development of bacteria/endotoxin translocation following thermal injury. The supplement of exogenous bifidobacteria could per se improve gut barriers, and attenuate bacteria/endotoxin translocation secondary to major burns.     

环氧合酶-2-1195G/A和锰超氧化物歧化酶9Ala/Val基因多态性与高脂饮食的交互作用及其与溃疡性结肠炎的关系

目的 探讨环氧合酶-2-1195G/A(COX-2-1195G/A)和锰超氧化物歧化酶9Ala/Val(MnSOD9Ala/Val)基因多态性与高脂饮食的交互作用及其与溃疡性结肠炎(UC)的关系。方法 采用病例-对照研究的方法,以750例UC患者及750例健康对照者的外周血白细胞为样本,采用聚合酶链反应(PCR)技术分析COX-2-1195G/A和MnSOD9Ala/Val基因多态性。结果 UC组和对照组COX-2-1195G/A(A/A)基因型的分布频率分别为49.07%和21.20%,MnSOD9Ala/Val(V/V)基因型的分布频率分别为50.13%和22.40%,差异均有统计学意义(P均＜0.01)。COX-2-1195G/A(A/A)基因型(OR=3.5808,95%CI=1.8062～5.3478)和MnSOD9Ala/Val(V/V)基因型(OR=3.4828,95%CI=1.9137～5.5496)者患UC的风险均显著增加。基因突变的协同分析结果显示,COX-2-1195G/A(A/A)/MnSOD9Ala/Val(V/V)基因型者在UC组和对照组中的分布频率分别为40.67%和8.40%,差异有统计学意义(P＜0.01),COX-2-1195G/A(A/A)/MnSOD9Ala/Val(V/V)基因型者患UC的风险显著增加(OR=7.5655,95%CI=4.1849～11.2037)。UC组高脂饮食率显著高于对照组(49.73%比20.13%,P＜0.01),高脂饮食与COX-2-1195G/A(A/A)(γ=11.81821)和 MnSOD9Ala/Val(V/V)(γ=9.0107)基因型均有交互作用。结论 COX-2-1195G/A(A/A)和MnSOD9Ala/Val(V/V)基因型及高脂饮食是UC的易患因素,基因多态性与高脂饮食的交互作用增加了UC的发病风险。     

儿童龋病与唾液CCL28 和sIgA 抗体的相关性

目的：探讨儿童唾液中趋化性细胞因子CCL28含量和sIgA 抗体分泌与儿童患龋状况之间的联系。方法： 抽取长沙市两所幼儿园3~5岁108名幼儿进行龋齿检查,根据检查结果分为无龋组[龋失补指数(dynamical mean-field theory,DMFT)=0];低龋组(DMFT=1~4);高龋组(DMFT≥5),收集非刺激性全唾液样本,用酶联免疫吸附试验 (ELISA) 定量检测唾液中 CCL28及sIgA的含量,并对结果进行统计学分析。结果：108名儿童唾液中CCL28的浓度为 (121.22±32.63) pg/mL,sIgA 的浓度为(16.49±8.02) μg/mL,两者之间存在线性正相关关系(r=0.734),且儿童唾液样本中 CCL28和sIgA含量随儿童患龋程度而增加(P<0.05)。结论：儿童龋患能够促进CCL28和sIgA分泌,且CCL28和sIgA的分 泌量与龋患程度呈正相关;CCL28对唾液中sIgA 的分泌起促进作用。     

《针灸逢源》耳病辨治规律探析

对《针灸逢源》中耳病相关内容进行整理分析,归纳该书耳病辨治规律。《针灸逢源》记载的耳病主要有耳聋、耳鸣、耵耳、聤耳、耳疮、耳衄,书中认为耳病由脏腑或经络异常导致,且有虚实之分。在治疗上以针灸为主,对耵耳、聤耳、耳衄还可运用中药治疗。取穴上阳经腧穴多于阴经腧穴,使用频次最多的穴位为听会穴,特定穴中五输穴使用频次较高,选穴分布以上肢部和头面颈项部为主。