大鼠骨骼肌挫伤后ASCT2 mRNA的表达变化

目的应用实时定量PCR技术检测大鼠骨骼肌挫伤后细胞中性氨基酸载体ASCT2 mRNA表达,分析其与挫伤时间的关系。方法建立大鼠骨骼肌挫伤模型,分别取伤后4、8、12、16、20、24、28、32 h及对照组检材。提取总RNA,逆转录合成第一链的cDNA,以RPL13为内参基因进行荧光定量检测,采用2-△△Ct法比较其与对照组肌肉组织中ASCT2 mRNA的相对表达量。结果损伤组肌肉组织中ASCT2 mRNA在挫伤后12、16h表达量分别为对照组的196.40%和189.15%,损伤后4、8、20、24、28、32h ASCT2 mRNA表达量与对照组相比无统计学差异(P>0.05),说明ASCT2 mRNA在损伤12～16h其表达量较正常组及其他损伤时间组要高,而在损伤20h以后ASCT2 mRNA表达量又降至正常水平。结论大鼠骨骼肌挫伤后32h之内ASCT2 mRNA的表达有一定的时间规律性,可望作为推断骨骼肌损伤时间的指标之一。     

猪瘟病毒NASBA-ELISA检测方法的建立与应用

为建立猪瘟病毒NASBA-ELISA检测方法,以猪瘟病毒(CSFV)E2基因为研究对象,利用Prim-er 5.0和Blastn生物软件,设计并合成了特异的核酸序列依赖扩增(NASBA)引物和探针。经条件优化,确定的最佳扩增反应条件为42℃,反应1.5h。临床检测结果显示,用该方法可特异地扩增CSFV E2基因约155bp的目的片段,而对TGEV、PRRSV、PCV2、PPV、PEDV、APP和PK-15正常细胞等核酸的检测均为阴性,可最低检出1×100～1×101 copies/μL,其灵敏度比RT-PCR略高。应用该方法对采自重庆部分地区猪场的125份样品进行了检测;结果,猪瘟病毒阳性检出率为5.6%,NASBA-ELISA法与RT-PCR法的检出符合率为94.4%。结果表明,NASBA-ELISA方法可应用于CSFV的快速鉴别检测。     

The relationship of methanol and formate concentrations in fatalities where methanol is detected

An automated headspace gas chromatography method was developed for the determination of formate (formic acid) in postmortem specimens, based on the in situ sulfuric acid-methanol methylation of formic acid to methyl formate. Diisopropyl ether was used as an internal standard. The method was applied to over 150 postmortem cases where methanol was detected. Of the 153 cases presented, 107 deaths were attributed to acute methanol toxicity. In the vast majority of the remaining 46 deaths, the methanol was determined to be present as a postmortem or perimortem artifact, or was otherwise incidental to the cause of death. Of the 76 victims who were found dead and blood was collected by the medical examiner, all but one had a postmortem blood formate concentration greater than 0.50 g/L (mean 0.85 g/L; n = 74). The sole exception involved suicidal ingestion of methanol where the blood methanol concentration was 7.9 g/L (790 mg/100 mL) and blood formate 0.12 g/L. In 97% (72/74) of the cases where blood was available, the blood formate was between 0.60 and 1.40 g/L. In 31 of the 153 cases, the victim was hospitalized and blood obtained on admission or soon after was analyzed for methanol and formate during the subsequent death investigation; the vast majority (27/30) had antemortem blood formate concentrations greater than 0.50 g/L. Cases with samples taken prior to death with blood formate concentrations less than 0.5 g/L can readily be explained by active treatment such as dialysis. The blood formate method has also been useful in confirming probable perimortem or postmortem contamination of one of more fluids or tissues with methanol (e.g., windshield washer fluid or embalming fluid), where methanol ingestion was unlikely.     

A Study of PCR Inhibition Mechanisms Using Real Time PCR*,†

Abstract: In this project, real time polymerase chain reaction (PCR) was utilized to study the mechanism of PCR inhibition through examination of the effect of amplicon length, melting temperature, and sequence. Specifically designed primers with three different amplicon lengths and three different melting temperatures were used to target a single homozygous allele in the HUMTH01 locus. The effect on amplification efficiency for each primer pair was determined by adding different concentrations of various PCR inhibitors to the reaction mixture. The results show that a variety of inhibition mechanisms can occur during the PCR process depending on the type of co‐extracted inhibitor. These include Taq inhibition, DNA template binding, and effects on reaction efficiency. In addition, some inhibitors appear to affect the reaction in more than one manner. Overall we find that amplicon size and melting temperature are important in some inhibition mechanisms and not in others and the key issue in understanding PCR inhibition is determining the identity of the interfering substance.     

Evaluation of Tamm‐Horsfall Protein and Uroplakin III for Forensic Identification of Urine

Abstract: In this study, Tamm‐Horsfall protein (THP), a major component of urinary protein, and uroplakin III (UPIII), a transmembrane protein widely regarded as a urothelium‐specific marker, were evaluated for forensic identification of urine by ELISA and/or immunohistochemistry. THP was detected in urine, but not in plasma, saliva, semen, vaginal fluid, or sweat by the simple ELISA method developed in this study. In addition, most aged urine stains showed positive results. The urine specificity of THP was confirmed by gene expression analysis. Therefore, as reported previously, ELISA detection of THP can be used as a presumptive test for urine identification. UPIII was specific for immunohistochemical staining of cells in centrifuged precipitate of urine. However, ELISA and RT‐PCR for UPIII were not specific for urine. UPIII may be applicable for forensic urine identification by immunohistochemistry.     

Characterization of the Products Formed by the Reaction of Trichlorocyanuric Acid with 2-Propanol

Abstract:  We report a recent investigation into the death of a cat that was initially thought to involve intentionally burning the animal via the use of an ignitable liquid. The exposure of the animal to flame was ruled out. Instead, forensic investigation revealed the intentional mixing together of a common outdoor swimming pool chlorinator, trichlorocyanuric acid (TCCA), and 2-propanol (aka, isopropyl alcohol or rubbing alcohol). The reaction of these two chemicals resulted in the formation of cyanuric acid residue, hydrochloric acid, and the evolution of a significant volume of chlorine gas. Further α-chlorination side reactions also occurred between 2-propanol and TCCA to produce a variety of chlorinated 2-propanone species that were detected on the submitted evidence. The identification of the products of both the main reaction and the side reactions allowed the authors to determine what chemicals were originally mixed together by the culprit.     

采用高效液相色谱-串联质谱法检测鲫鱼组织3-甲基喹(噁)啉-2-羧酸残留量

将待测鱼肉和鱼皮匀浆样品经碱水解、液-液萃取后,采用HPLC-TMS方法对其中的3-甲基喹(噁)啉-2-羧酸(MQCA)残留量进行了检测.结果,此方法的最低检测限为1.16μg/kg,市售鲫鱼样品中MQCA的含量低于检测限.结果表明,HPLC-TMC灵敏,易操作,能满足鱼组织内MQCA残留量检测的要求.     

一种改良酸性磷酸酶试验鉴定精斑的方法

本文报道了一种改良的酸性磷酸酶试验方法。用该法鉴定精斑，可克服非特异性反应，排除其它体液和植物汁液的影响。     

宣白承气汤治疗肺炎致脓毒症42例临床观察

目的 观察宣白承气汤对肺炎所致脓毒症的临床疗效,并探究其作用机制。方法 将84例肺炎致脓毒症患者随机分为对照组和治疗组,每组42例,两组患者均给予西医常规治疗,治疗组在常规治疗的基础上加用宣白承气汤,比较两组患者治疗前后血乳酸（lactate acid,Lac）浓度、上腔静脉血氧饱和度（superior vena cava oxygen saturation,ScvO2）、降钙素原（procalcitonin,PCT）水平及社区获得性肺炎评分量表（confusion, urea, respiratory rate, blood pressure, age≥65,CURB-65）、序贯器官衰竭评估量表（sequential organ failure assessment,SOFA）评分。结果 与治疗前比较,两组患者治疗后Lac、PCT均显著降低（P＜0.05）,ScvO2水平均显著升高（P＜0.05）;治疗组患者Lac降低程度和ScvO2升高程度显著大于对照组（P＜0.05）。与治疗前比较,两组患者治疗后CURB-65评分、SOFA评分均显著降低（P＜0.05）,且治疗组患者CURB-65、SOFA评分降低程度显著大于对照组（P＜0.05）。结论 宣白承气汤能改善肺炎所致脓毒症患者的灌注指标,减轻炎症反应,效果显著。     

茵栀黄口服液联合西药治疗妊娠期肝内胆汁淤积症的疗效及母婴结局分析

目的 观察中西医结合疗法对妊娠期肝内胆汁淤积症的疗效及母婴结局的影响。方法 将60例妊娠期肝内胆汁淤积症患者随机分为观察组和对照组,每组30例。对照组给予熊去氧胆酸、腺苷蛋氨酸及还原型谷胱甘肽等西药常规治疗,观察组在对照组基础上加用茵栀黄口服液。分别检测两组患者血清总胆汁酸（total bile acid,TBA）、丙氨酸氨基转移酶（alanine aminotransferase,ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase,AST）、总胆红素（total bilirubin,TBIL）水平,对两组患者瘙痒症状进行评分,观察其临床疗效,并对母婴结局进行分析。结果 治疗后两组患者血清TBA、ALT、AST及TBIL水平均明显下降,差异有统计学意义（P<0.05）;且两组患者治疗前后TBIL差值比较,差异有统计学意义（P<0.05）。治疗后两组患者瘙痒症状评分均明显减少（P<0.05）,且观察组瘙痒症状疗效优于对照组（P<0.05）。观察组可明显延长胎儿孕周,且产后24 h出血量、发生早产率及转入新生儿病房率明显低于对照组（P<0.05）。结论 中西医结合方法治疗妊娠期肝内胆汁淤积症具有较好的临床疗效,可明显降低围产期母婴风险。