1.RNA-Seq for pathogenesis of Candida albicans
Jianchai LIU ; Huanzhang LIU ; Yanwei LIU ; Jinkun YAN ; Heping ZHANG ; Jingliang SU
Chinese Journal of Zoonoses 2017;33(1):72-80
In the paper,we introduced the peculiarity of Candida albicans and the disease caused by it,expounded the complexity of the pathogenesis,enumerated the advantages of the RNA-Seq and reviewed its application to study on the pathogenesis of Candida albicans,found out some shortages in previous studies,and anticipated the possible trends of such study in future.In conclusion,some remarkable achievements will bring about by use of improved RNA-Seq for intensive researches on the pathogenesis of Candida albicans.
2.Alterations in nasal microbiota of patients with amyotrophic lateral sclerosis.
Kaixiong LIU ; Qifu GUO ; Ying DING ; Li LUO ; Jianchai HUANG ; Qijie ZHANG
Chinese Medical Journal 2024;137(2):162-171
BACKGROUND:
Links between alterations in gut microbiota composition and amyotrophic lateral sclerosis (ALS) have previously been reported. This study aimed to examine the microbiota in the nasal cavity of ALS.
METHODS:
Sixty-six ALS patients and 40 healthy caregivers who live in close proximity with patients were enrolled. High throughput metagenomic sequencing of the 16S ribosomal deoxyribonucleic acid (rDNA) gene V3-V4 region of nasal microbiota was used to characterize the alpha and beta diversity and relative abundance of bacterial taxa, predict function, and conduct correlation analysis between specific taxa and clinical features.
RESULTS:
The nasal microbiome of ALS patients showed lower alpha diversity than that of corresponding healthy family members. Genera Gaiella , Sphingomonas , Polaribacter _1, Lachnospiraceae _NK4A136_group, Klebsiella , and Alistipes were differentially enriched in ALS patients compared to controls. Nasal microbiota composition in ALS patients significantly differed from that in healthy subjects (unweighted UniFrac P = 0.001), while Linear discriminant analysis Effect Size (LEfSe) analysis indicated that Bacteroidetes and Firmicutes dominated healthy nasal communities at the phylum level, whereas Actinobacteria was the predominant phylum and Thermoleophilia was the predominant class in ALS patients. Genus Faecalibacterium and Alistipes were positively correlated with ALS functional rating scale revised (ALSFRS-R; rs = 0.349, P = 0.020 and rs = 0.393, P = 0.008), while Prevotella -9 and Bacteroides operational taxonomic units (OTUs) were positively associated with lung function (FVC) in ALS patients ( rs = 0.304, P = 0.045, and rs = 0.300, P = 0.048, respectively). Prevotella -1 was positively correlated with white blood cell counts (WBC, rs = 0.347, P = 0.021), neutrophil percentage (Neu%, rs = 0.428, P = 0.004), and neutrophil-to-lymphocyte ratio (NLR, rs = 0.411, P = 0.006), but negatively correlated with lymphocyte percentage (Lym%, rs = -0.408, P = 0.006). In contrast, Streptococcus was negatively associated with Neu% ( rs = -0.445, P = 0.003) and NLR ( rs = -0.436, P = 0.003), while positively associated with Lym% ( rs = 0.437, P = 0.003). No significant differences in nasal microbiota richness and evenness were detected among the severe and mild ALS patients.
CONCLUSIONS
ALS is accompanied by altered nasal microbial community composition and diversity. The findings presented here highlight the need to understand how dysbiosis of nasal microbiota may contribute to the development of ALS.
Humans
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Amyotrophic Lateral Sclerosis/microbiology*
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Feces/microbiology*
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Microbiota/genetics*
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Gastrointestinal Microbiome/genetics*
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Bacteria/genetics*
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RNA, Ribosomal, 16S/genetics*
3.Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein.
Shuai DONG ; Weiqin MENG ; Ling MO ; Jinlong CHEN ; Jingnan SHI ; Zhe YANG ; Tong LI ; Qianqian XU ; Zhiqiang SHEN ; Jianchai LIU ; Jinliang WANG
Chinese Journal of Biotechnology 2023;39(12):4915-4926
A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.
Animals
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Mice
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Peste-des-Petits-Ruminants/prevention & control*
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Antibodies, Monoclonal
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Reproducibility of Results
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Peste-des-petits-ruminants virus
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Antibodies, Viral
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Enzyme-Linked Immunosorbent Assay
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Goats