1.Evaluation of ITS region sequencing strategy for the identification of the pathogenic fungi from fungal sinusitis tissues
Xinxin LU ; Jiajing GENG ; Yunchuan LI ; Bing ZHOU ; Liang WAN ; Xiangdong WANG ; Zi ZHANG ; Demin HAN
Chinese Journal of Laboratory Medicine 2010;33(2):126-131
Objective To establish a molecular technique of internal transcribed spacer (ITS) sequencing to identify pathogenic fungi species from the fungal sinusitis tissues. Methods Total 270 sinusitis tissues samples were collected by endoscopic surgery from 2006 to 2008. The histopathology, organize spring clip culturation and ITS region (ITS region region of fungal rRNA, including ITS1-5. 8S rRNA-ITS2) sequencing were employed simultaneously. And then to evaluate the ITS sequencing as the tool for identification of pathogenic fungi directly from clinical samples. Results Of the 270 samples, histopathology positive rate was 80.0% (216/270) , organize spring clip positive rate was 80.0% (216/ 270), fungal culturation positive rate was 53.0% (143/270) , ITS region sequencing positive rate was 63. 0% [ (134 +28 +8)/270], There were 22 species and 6 genera identified by fungal culturation, and 32 species identified by ITS region sequencing. Conclusion ITS region sequencing will become a applicable tool in clinical laboratory in future.
2.Autism-like Behaviors in Male Juvenile Offspring after Maternal Glyphosate Exposure
Yaoyu PU ; Li MA ; Jiajing SHAN ; Xiayun WAN ; Bruce D. HAMMOCK ; Kenji HASHIMOTO
Clinical Psychopharmacology and Neuroscience 2021;19(3):554-558
Objective:
Exposure to the herbicide glyphosate during pregnancy and lactation may increase the risk for autism spectrum disorder (ASD) in offspring. Recently, we reported that maternal exposure of formulated glyphosate caused ASD-like behaviors in juvenile offspring. Here, we investigated whether maternal exposure of pure glyphosate could cause ASD-like behaviors in juvenile offspring.
Methods:
Water or 0.098% glyphosate was administered as drinking water from E5 to P21 (weaning). Behavioral tests such as grooming test and three-chamber social interaction test in male offspring were performed from P28 to P35.
Results:
Male offspring showed ASD-like behavioral abnormalities (i.e., increasing grooming behavior and social interaction deficit) after maternal exposure of glyphosate.
Conclusion
The findings suggest that the exposure of glyphosate during pregnancy and lactation may cause ASD-like behavioral abnormalities in male juvenile offspring. It is likely that glyphosate itself, but not the other ingredients, may contribute to ASD-like behavioral abnormalities in juvenile offspring.
3.Autism-like Behaviors in Male Juvenile Offspring after Maternal Glyphosate Exposure
Yaoyu PU ; Li MA ; Jiajing SHAN ; Xiayun WAN ; Bruce D. HAMMOCK ; Kenji HASHIMOTO
Clinical Psychopharmacology and Neuroscience 2021;19(3):554-558
Objective:
Exposure to the herbicide glyphosate during pregnancy and lactation may increase the risk for autism spectrum disorder (ASD) in offspring. Recently, we reported that maternal exposure of formulated glyphosate caused ASD-like behaviors in juvenile offspring. Here, we investigated whether maternal exposure of pure glyphosate could cause ASD-like behaviors in juvenile offspring.
Methods:
Water or 0.098% glyphosate was administered as drinking water from E5 to P21 (weaning). Behavioral tests such as grooming test and three-chamber social interaction test in male offspring were performed from P28 to P35.
Results:
Male offspring showed ASD-like behavioral abnormalities (i.e., increasing grooming behavior and social interaction deficit) after maternal exposure of glyphosate.
Conclusion
The findings suggest that the exposure of glyphosate during pregnancy and lactation may cause ASD-like behavioral abnormalities in male juvenile offspring. It is likely that glyphosate itself, but not the other ingredients, may contribute to ASD-like behavioral abnormalities in juvenile offspring.
4.A Study on the Characteristics of Articulation Movement in Patients with Dysarthria under Different Vowel Tasks
Jiamei CHEN ; Qin WAN ; Jiajing LIANG ; Fenfen HUI ; Hongfen QIAO ; Wenzhuo SHU
Journal of Audiology and Speech Pathology 2024;32(5):413-417
Objective To investigate the acoustic characteristics of monophthong vowels and compound vow-els articulation movement in patients with dysarthria.Methods A total of 24 patients aged 40~81 years old with dysarthria from hospitals,and 24 healthy adults aged 40~80 years old from communities in Shanghai were recrui-ted.The first and second formants(F1,F2)of monophthong vowels/a/,/i/,/u/were extracted from the speech samples,and mandibular distance,tongue distance,vowel space area(VSA)and vowel articulation index(VAI)were calculated.The slope of the second formant of compound vowels/ai/,/ua/,/uai/was extracted and calculat-ed.The acoustic parameters of two groups of subjects were compared under different tasks.Results Under the monophthong vowels task,F1 of/a/,F2 of/i/,mandibular distance,tongue distance,VSA and VAI in the pa-tients were significantly lower than those in the control group(P<0.01).F2 of/u/was significantly higher than that in the control group(P<0.01).F2 slope of/ai/,/ua/,/uai/was significantly lower than that of the control group(P<0.05).Conclusion The space of monophthong vowels articulation movement was reduced in the pa-tients with dysarthria,presenting a state of aggregation.The speed of compound vowels articulation movement was decreased.
5.Construction and preliminary phenotype analysis of mice with Retnlb knockout in colon cells
Feiying WANG ; Lingling ZHOU ; Beibei CHENG ; Jiajing WAN ; Chao ZHANG ; Jian YI ; Lan SONG ; Aiguo DAI
Chinese Journal of Pathophysiology 2024;40(10):1788-1796
AIM:This study utilized CRISPR/Cas9 technology to create Retnlb floxp knock-in mice,followed by the application of the Cre-LoxP recombination system to generate intestinal epithelial-specific Retnlb gene knockout mice(Retnlb-CKO).This model was developed to investigate the pathogenic mechanisms of Retnlb in inflammatory bowel disease.METHODS:Female and male C57BL/6N mice,aged 8 weeks with the Retnlbflox/+genotype,were housed togeth-er for breeding.Offsprings were screened to identify those with the Retnlbflox/flox genotype.These mice were then crossed with Vil1-Cre transgenic mice,which express Cre recombinase specifically in intestinal epithelial cells,resulting in Retnlb-flox/+,Cre+mice.Subsequent crosses between Retnlbflox/+,Cre+mice and Retnlbflox/flox mice produced Retnlbflox/flox,Cre+mice(Retnlb-CKO).Six 8-week-old Retnlbflox/flox,Cre+mice and their littermate Retnlbflox/flox mice were selected for experiments.RT-qPCR and immunohistochemistry were used to assess Retnlb mRNA and protein levels in colonic epithelium.Phenotypic observa-tions included body length,weight,diet,and reproductive capability.Tissue-to-body weight ratios were calculated to ana-lyze growth and development.Intestinal barrier integrity and colonic expression of inflammatory factors were evaluated.RESULTS:The conditional gene knockout mouse model with specific deletion of Retnlb in intestinal epithelial cells was successfully established and validated through genetic identification,mRNA and protein analysis.Compared to Retnlbflox/flox mice,Retnlb-CKO mice exhibited no significant differences in body length,weight,diet,or reproductive capability.There were no differences in the ratios of heart,liver,spleen,lung,kidney,and colon weight to body weight,nor were there morphological differences in various tissues.However,the mRNA expression of tight junction proteins ZO-1,Occlu-din,and Claudin3 in colon tissues of Retnlb-CKO mice was significantly reduced(P<0.01).PAS staining and immunohis-tochemistry revealed a significant decrease in the number of goblet cells and lysozyme-positive cells in the colon tissues of Retnlb-CKO mice(P<0.01).HE staining showed no obvious pathological change in colon tissues of Retnlb-CKO mice.RT-qPCR further demonstrated a significant downregulation of pro-inflammatory factors NLRP3,interleukin-6(IL-6),IL-1β,and tumor necrosis factor-α(TNF-α)in colon tissues(P<0.01),along with significant downregulation of inflamma-tion signaling pathway proteins TLR4,MyD88,and NF-κB(P<0.01).CONCLUSION:A conditional colon epithelial cell Retnlb gene knockout mouse model was successfully constructed and validated.The absence of Retnlb in colon cells led to impaired intestinal barrier function,decreased mRNA expression of pro-inflammatory factors in colon tissue,and downregulation of mRNA expression of inflammatory pathway proteins TLR4,MyD88,and NF-κB.