Objective To compare the sensitivity and specificity of different fluorescent quantitation polymerase chain reaction(PCR) for genes detection of Streptococcus pneumonia.Methods By designing five sets of fluorescent quantitation PCR primers targeting different genes of Streptococcus pneumonia (lytA,ply,spn9802,psaA,cspA),the sensitivity and specificity of fluorescent quantitation PCR was determined through detecting 37 strains of Streptococcus pneumonia and 28 strains of non-Streptococcus pneumonia,as well as 80 sputum specimens from suspected pneumonia cases.Results These five primers had excellent sensitivity with 90％ amplification efficiency through analysis of the standard curve.The change of Ct value was less than 0.5,indicating that these five primers have good stability.By detecting 37 strains of Streptococcus pneumonia and 28 strains of non-Streptococcus pneumonia,it showed that the specificity of lytA,ply and spn9802 was better than the other two primers (positive rate and negative rate was all 100％).But the detection limit of lytA,ply was one order of magnitude than spn9802 (101 CFU/ml vs.102 CFU/ml).The specificity of psaA and cspA was worse [negative rate of psaA was 89％ (25/28),positive rate of cspA was 97％ (36/37)].By detecting 80 sputum specimens from suspected pneumonia cases,it showed that the specificity of fluorescent quantitation PCR were better than the bacterial culture.The specificity of lytA and ply was the best two [positive rate:92.50％(74/80),90.00％(72/80)],and next was spn9802 [86.25％(69/80)],the specificity ofpsaA and cspA was the worst two for detection [76.25％(61/80) and 78.75％(63/80)].Conclusions The sensitivity and specificity of fluorescent quantitation PCR genes used in clinic for detecting Streptococcus pneumonia is different.LytA and ply are the best primers in specificity,next is spn9802,and psaA and cspA are the worst primers for in specificity detection.

Objective:To summarize the risk factors, clinical manifestations, diagnosis and treatment of limb thrombosis in neonates.Methods:The clinical data of 14 neonates with limb thrombosis were hospitalized in neonatology department at Shengjing Hospital of China Medical University from February 2012 to February 2022 were retrospectively analyzed.Results:All the 14 cases of limb thrombosis were premature infants, with an average gestational age of 29 weeks and 5 days(27 weeks and 3 days to 33 weeks and 1 day), including eight cases of arterial embolism and six cases of venous embolism.Among them, 13(92.9%) cases were diagnosed with infectious diseases such as septicemia or neonatal necrotizing enterocolitis within 48 hours before embolization, and all had a history of peripheral arterial and venous catheterization.During the early stage of embolization, limb artery embolism was characterized by weakened distal artery pulsation, pale skin, gradual cyanosis and even gangrene.Limb venous embolism was manifested as limb swelling, skin congestion and cyanosis, but the arterial pulsation was normal.Fourteen cases were confirmed by vascular ultrasound.All the eight cases with arterial embolization were treated with heparin anticoagulation, five of which were cured, with an effective rate of 62.5%.The average time of heparin use in five cases was 2.5 days(2-3 days). One patient was not effective after 2 days of heparin treatment, and recovered after thrombectomy.Another two cases had distal limb gangrene, and them were treated with heparin for 5 days and 7 days.All of the six cases with venous embolism were cured, of which four cases were treated with heparin for an average of 8.5 days(4-15 days), and the other two cases were cured after general treatment.There were no bleeding events in the 12 infants treated with heparin.Conclusion:Peripheral arterial and venous catheterization during infection of preterm infants is the most common cause of limb thrombosis.The smaller body weight and gestational age, the thinner blood vessels, the higher risk of occurrence.Vascular ultrasound is the most commonly used examination method for neonatal thrombosis, and heparin anticoagulant therapy is the most commonly used treatment measure.When the treatment effect of heparin is not good, other treatment methods should be sought.

Clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9),a cluster of regularly spaced short palindromic repeats,is a natural immune defense system for bacteria and archaea to identify themselves and exogenous invading DNA fragments,protecting them from viruses.In recent years,CRISPR/Cas9 has become a revolutionary gene editing tool.Its specific targeted spot-cutting ability also plays an important role in nucleic acid detection,bacterial typing,etc.,and has shown great application potential in the field of medical testing.Based on the latest researches,this paper reviews the progress of CRISPR/Cas9 application in the new techniques of nucleic acid detection,pathogen typing and bacterial evolution in laboratory medicine,and also summarizes the application prospect of CRISPR technology in the field of laboratory medicine.

Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin (CDN), the major active ingredient of the traditional Chinese medicinal herb , has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-B-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner. Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1 production induced by LPS , which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.

[This corrects the article DOI: 10.1016/j.apsb.2019.02.003.].

Psoraleae Fructus (PF) is a well-known traditional herbal medicine in China, and it is widely used for osteoporosis, vitiligo, and other diseases in clinical settings. However, liver injury caused by PF and its preparations has been frequently reported in recent years. Our previous studies have demonstrated that PF could cause idiosyncratic drug-induced liver injury (IDILI), but the mechanism underlying its hepatotoxicity remains unclear. This paper reports that bavachin isolated from PF enhances the specific stimuli-induced activation of the NLRP3 inflammasome and leads to hepatotoxicity. Bavachin boosts the secretion of IL-1β and caspase-1 caused by ATP or nigericin but not those induced by poly(I:C), monosodium urate crystal, or intracellular lipopolysaccharide. Bavachin does not affect AIM2 or NLRC4 inflammasome activation. Mechanistically, bavachin specifically increases the production of nigericin-induced mitochondrial reactive oxygen species among the most important upstream events in the activation of the NLRP3 inflammasome. Bavachin increases the levels of aspartate transaminase and alanine aminotransferase in serum and hepatocyte injury accompanied by the secretion of IL-1β via a mouse model of lipopolysaccharide-mediated susceptibility to IDILI. These results suggest that bavachin specifically enhances the ATP- or nigericin-induced activation of the NLRP3 inflammasome. Bavachin also potentially contributes to PF-induced idiosyncratic hepatotoxicity. Moreover, bavachin and PF should be evaded among patients with diseases linked to the ATP- or nigericin-mediated activation of the NLRP3 inflammasome, which may be a dangerous factor for liver injury.
Adenosine Triphosphate ; Animals ; Chemical and Drug Induced Liver Injury/etiology* ; Flavonoids ; Humans ; Inflammasomes ; Mice ; NLR Family, Pyrin Domain-Containing 3 Protein ; Nigericin