微滴式数字PCR和实时荧光定量PCR对新型冠状病毒肺炎患者不同时间血便尿中核酸检测结果初步探讨

目的 分析微滴式数字PCR(droplet digital PCR, ddPCR)和实时荧光定量PCR(quantitative real-time PCR,qPCR)的核酸检测结果,比较两种方法检测各类样本的差异性,为改进新型冠状病毒核酸检测方案提供数据支持。 方法 利用ddPCR和qPCR技术对已经确诊的3例新型冠状病毒肺炎患者发病不同时间的全血、尿液、粪便共22份标本进行新型冠状病毒核酸检测。 结果 两种方法对人保守区域基因扩增结果一致:全血标本信号最强,尿液次之,粪便最少;ddPCR在1份全血,1份尿液,5份粪便中检出ORF-1ab和N基因的阳性微滴,qPCR仅在3份粪便中检出上述基因,漏检的3个标本基因拷贝数平均浓度为128 copies/ml;ddPCR在发病<5、5~15、>15 d的各类标本中都有检出,qPCR检出以中晚期为主;重症病例用ddPCR均可测到阳性微滴,qPCR检测的各类标本均为阴性;轻症病例的各类标本中qPCR只有粪便核酸检测阳性,ddPCR检出率高于qPCR。 结论 ddPCR可以有效克服qPCR 灵敏度不足的难题,是对qPCR 的有益补充,尤其是针对病毒载量比较低的血液、尿液和可疑的粪便或肛拭子标本,适用于早期感染的判断及患者治愈后出院诊断。     

猪粪饲养家蝇幼虫的营养成分研究

目的 :分析猪粪饲养家蝇 (Muscadomestica)幼虫的营养成分。方法 :引种野生家蝇 ,用猪粪饲养幼虫 ,收获 3期老熟幼虫 ,测定粗蛋白、粗脂肪、氨基酸和脂肪酸构成及微量元素含量 ,用粮食与农业组织 世界卫生组织 (FAO WHO)提出的参考蛋白模式评价 ,并与麦麸饲养的幼虫及鱼粉比较。结果 :每公斤猪粪平均可产(0 13± 0 0 4)～ (0 2 0± 0 12 )kg鲜蛆 ,鲜蛆烘干比为 1∶0 31± 0 12 ,粗蛋白 42 36 %～ 5 2 33 % ,必须氨基酸含量占总氨基酸的 45 10 % (E % ) ,必须氨基酸与非必需氨基酸总量的比值 (E/N)为 0 82 ,E % /T为 3 0 1,脂肪酸含量为 13 73 %～ 16 83 % ,油酸、亚油酸及亚麻酸等必需脂肪酸分别为 36 99% ,7 6 1%和 0 35 % ,Fe 2 91 5mg/kg ,Cu 87 5 5mg/kg ,Zn 2 46 8mg/kg ,Mn 2 49 45mg/kg ,比麦麸饲养幼虫更接近参考蛋白模式 ,营养价值优于鱼粉。结论 :猪粪饲养的家蝇幼虫比麦麸饲养的幼虫更接近参考蛋白模式 ,营养价值优于鱼粉     

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??Objective??To detect the level of fecal primary and secondary bile acids in infants with infantile cholestatic hepatopathy??ICH??and analyze its clinical value. Methods??Thirty infants with ICH were enrolled in this study??who were diagnosed with infantile cholestatic hepatopathy. Thirty infants with good health condition were enrolled as the healthy control group. The fecal samples were collected respectively in the preparatory treatment phase and treatment phase from infants with ICH and from the healthy infants. Bile acids were extracted from infants’ feces and were quantitatively analyzed by liquid chromatography-mass spectroscopy. Results??Among the fecal primary bile acids??the level of cholic acid??chenodeoxycholic and glycochenodeoxycholic acid both in the ICH preparatory treatment group and ICH treatment group was significantly lower than that in the healthy control group??P??0.016??.The level of fecal cholic acid and chenodeoxycholic acid of ICH treatment group was higher than in the ICH preparatory treatment group??P??0.016??. Among the fecal secondary bile acids??the level of lithocholic acid both in the ICH preparatory treatment group and ICH treatment group was significantly lower than that in the healthy control group??P??0.016????and the level of ursodeoxycholic acid in the ICH preparatory treatment group was lower than that in the ICH treatment group and healthy control group??P??0.016??. Conclusion??In infants with ICH, the changes of fecal primary bile acids and fecal secondary bile acids have their own characteristics at the early stage of treatment, which may be caused by the short-term treatment, the prognosis of the disease itself and the changes of intestinal function, including intestinal bacteria. Clinical attention should be paid to these changes.     

Occurrence of a mosquito vector in bird houses: Developmental consequences and potential epidemiological implications

Even with continuous vector control, dengue is still a growing threat to public health in Southeast Asia. Main causes comprise difficulties in identifying productive breeding sites and inappropriate targeted chemical interventions. In this region, rural families keep live birds in backyards and dengue mosquitoes have been reported in containers in the cages. To focus on this particular breeding site, we examined the capacity of bird fecal matter (BFM) from the spotted dove, to support Aedes albopictus larval growth. The impact of BFM larval uptake on some adult fitness traits influencing vectorial capacity was also investigated. In serial bioassays involving a high and low larval density (HD and LD), BFM and larval standard food (LSF) affected differently larval development. At HD, development was longer in the BFM environment. There were no appreciable mortality differences between the two treatments, which resulted in similar pupation and adult emergence successes. BFM treatment produced a better gender balance. There were comparable levels of blood uptake and egg production in BFM and LSF females at LD; that was not the case for the HD one, which resulted in bigger adults. BFM and LSF females displayed equivalent lifespans; in males, this parameter was shorter in those derived from the BFM/LD treatment. Taken together these results suggest that bird defecations successfully support the development of Ae. albopictus. Due to their cryptic aspects, containers used to supply water to encaged birds may not have been targeted by chemical interventions.     

粪便内中短链脂肪酸柱前衍生化UHPLC-QTOF-MS/MS分析方法研究

建立柱前衍生化超高效液相色谱-串联四级杆飞行时间质谱(UHPLC-QTOF-MS/MS)定性、定量表征小鼠粪便内中短链脂肪酸的方法,并用于评价其在抗生素处置前后小鼠粪便样品中的变化。动物实验方案获得江苏省中医药研究院动物实验伦理委员会的批准。以3-硝基苯肼为衍生化试剂,对衍生化反应条件、色谱及质谱条件进行优化,建立了粪便中16个中短链脂肪酸的定性、定量方法。所有分析物在各自浓度范围内线性关系良好(R2>0.99),日内、日间精密度RSD均小于10%,重复性RSD小于6%,加样回收率介于80%~120%之间,样品36 h内稳定性RSD小于7%;联合抗生素处置后小鼠粪便内的中短链脂肪酸种类和含量均发生了明显改变。其中,甲酸、乙酸、丙酸、丁酸、异丁酸、戊酸、乳酸等含量明显降低,而庚酸和丁二酸的含量明显增加。该方法准确、可靠,可作为粪便内中短链脂肪酸的表征方法。     

Pathogens transmitted in animal feces in low- and middle-income countries

Animals found in close proximity to humans in low-and middle-income countries (LMICs) harbor many pathogens capable of infecting humans, transmissible via their feces. Contact with animal feces poses a currently unquantified—though likely substantial—risk to human health. In LMIC settings, human exposure to animal feces may explain some of the limited success of recent water, sanitation, and hygiene interventions that have focused on limiting exposure to human excreta, with less attention to containing animal feces.We conducted a review to identify pathogens that may substantially contribute to the global burden of disease in humans through their spread in animal feces in the domestic environment in LMICs. Of the 65 potentially pathogenic organisms considered, 15 were deemed relevant, based on burden of disease and potential for zoonotic transmission. Of these, five were considered of highest concern based on a substantial burden of disease for which transmission in animal feces is potentially important: Campylobacter, non-typhoidal Salmonella (NTS), Lassa virus, Cryptosporidium, and Toxoplasma gondii. Most of these have a wide range of animal hosts, except Lassa virus, which is spread through the feces of rats indigenous to sub-Saharan Africa. Combined, these five pathogens cause close to one million deaths annually. More than half of these deaths are attributed to invasive NTS. We do not estimate an overall burden of disease from improperly managed animal feces in LMICs, because it is unknown what proportion of illnesses caused by these pathogens can be attributed to contact with animal feces.Typical water quantity, water quality, and handwashing interventions promoted in public health and development address transmission routes for both human and animal feces; however, sanitation interventions typically focus on containing human waste, often neglecting the residual burden of disease from pathogens transmitted via animal feces. This review compiles evidence on which pathogens may contribute to the burden of disease through transmission in animal feces; these data will help prioritize intervention types and regions that could most benefit from interventions aimed at reducing human contact with animal feces.     

栀子苷对大鼠肝损伤及血清、肝脏、粪便中胆汁酸的影响

目的 探讨不同剂量栀子苷对大鼠肝脏的毒性,以及对其血清、肝组织及粪便中胆汁酸的影响。方法 60只SD大鼠,雌雄各半,按体质量随机分成空白组及栀子苷（50,100,200,400 mg·kg^-1）组,共5组,每组12只,每天灌胃给药1次,连续7 d,于第8天收集大鼠血清、肝组织和盲肠内容物。检测各组大鼠血清丙氨酸氨基转移酶（ALT）,门冬氨酸氨基转移酶（AST）,碱性磷酸酶（ALP）活性,白蛋白（ALB）,总胆红素（TBIL）,总胆汁酸（TBA）,肌酐（Crea）,尿素（Urea）含量,肝脏组织切片分别作苏木素-伊红（HE）染色,免疫组化检测细胞角蛋白7（CK7）,细胞角蛋白19（CK19）蛋白表达,蛋白免疫印迹法（Western blot）测定肝组织CK7,CK19蛋白表达,实时荧光定量聚合酶链式反应（Real-time PCR）检测胆固醇7α-羟化酶（CYP7A1）,胆固醇27α-羟化酶（CYP27A1）,胆固醇12α-羟化酶（CYP8B1） mRNA表达情况,采用超高效液相色谱-串联质谱（UPLC-MS）测定血清、肝脏及盲肠内容物中18种胆汁酸含量。结果 与空白组比较,各剂量栀子苷组TBIL水平显著升高（P<0.01）;400 mg·kg^-1栀子苷组ALT,AST活性和TBA含量明显升高（P<0.05,P<0.01）;HE染色表明,与空白组比较,栀子苷200,400 mg·kg^-1组肝组织的汇管区有胆管反应,胆管周围有炎症细胞的浸润,尤以栀子苷400 mg·kg^-1毒性明显。栀子苷400 mg·kg^-1栀子苷组肝组织中CK7,CK19表达较空白组明显增加（P<0.05,P<0.01）。与空白组比较,400 mg·kg^-1栀子苷组肝组织中甘氨熊脱氧胆酸（GUDCA）,甘氨猪去氧胆酸（GHDCA）含量明显下降（P<0.05,P<0.01）,牛磺鹅去氧胆酸（TCDCA）,猪去氧胆酸（HDCA）,胆酸（CA）,鹅去氧胆酸（CDCA）含量显著升高（P<0.01）,甘氨胆酸（GCA）,甘氨鹅去氧胆酸（GCDCA）,甘氨脱氧胆酸（GDCA）,甘氨石胆酸（GLCA）,牛磺熊去氧胆酸（TUDCA）,GUDCA,GHDCA,熊去氧胆酸（UDCA）,牛磺石胆酸（TLCA）的含量占比降低,TCDCA,HDCA,CA,CDCA,脱氧胆酸（DCA）的占比增加;血清中GHDCA,石胆酸（LCA）含量显著下降（P<0.01）,牛磺猪去氧胆酸（THDCA）,牛磺胆酸（TCA）,GCA,TCDCA,UDCA,CA,CDCA,DCA含量明显增加（P<0.05,P<0.01）,CA/DCA显著升高（P<0.01）,且CA与CDCA含量占比分别上升19.60%和4.63%;与空白组比较,400 mg·kg^-1栀子苷组盲肠内容物中所有的胆汁酸含量均下降,其中GCA,UDCA,HDCA,GCDCA,GDCA,TLCA,GLCA,CDCA,DCA,LCA含量明显下降（P<0.05,P<0.01）。Real-time PCR结果显示,与空白组比较,400 mg·kg^-1栀子苷组肝组织中CYP7A1,CYP27A1 mRNA表达明显增加（P<0.05,P<0.01）。结论 400 mg·kg^-1栀子苷灌胃,可导致大鼠明显的肝损伤,其肝脏、血清与粪便中胆汁酸发生显著改变,且同一种胆汁酸在肝脏、血清与粪便中的变化不尽相同。然而,由栀子苷所导致的肝损伤与体内胆汁酸变化间的因果关系尚不清楚,仍有待后续进一步研究给予阐明。     

茵栀黄颗粒联合抚触护理治疗新生儿黄疸30例疗效观察

目的观察茵栀黄颗粒联合抚触护理治疗新生儿黄疸的疗效。方法选取2012年6月至2013年12月来我院就诊的新生儿黄疸60例,依据分层随机分组方法将患儿分为治疗组与对照组,每组30例。对照组给予茵栀黄颗粒冲服,治疗组在对照组的基础上联合抚触护理,2组同时接受常规治疗,均治疗7d。比较2组患儿治疗前及治疗7d后的血清胆红素水平、胆红素下降幅度、黄疸消退时间、胎粪第1次变黄时间及2组疗效。结果治疗前2组患儿指标均未见明显差异（P〉0．05）,治疗7d后,治疗组患儿的血清胆红素水平及黄疸消退时间均明显低于对照组,而治疗组的胎粪第1次变黄时间明显短于对照组,治疗组胆红素下降幅度则明显大于对照组,同时观察组患儿的疗效明显优于对照组（P〈0．05）。结论茵栀黄颗粒联合抚触护理对于新生儿黄疸患儿具有更好地降低血清胆红素及促进黄疸症状消退的功效,值得临床推广使用。     

麻仁滋脾丸和益寿通通便作用研究

麻仁滋脾丸对小鼠有一定通便作用，可增加鼠粪水分含量，并有一定促进大鼠大肠推进运动作用。济川煎化裁方益寿通则有强的通便效果，其通便机理可能与增加鼠粪水分含量，促进小肠和大肠推进运动有关。