2.Color Doppler ultrasonography in evaluation on renal hemodynamiacs in perioperation of percutaneous nephrolithotomy
Li LIU ; Minhui QIN ; Liang MU ; Shen LI ; Aihua WEI
Chinese Journal of Interventional Imaging and Therapy 2011;08(5):371-375
ObjectiveTo evaluate the impacts of pereutaneous nephrolithotomy (PCNL) on perioperative renal hemodynamics.MethodsThe hemodynamics of operated renal arteries of 30 patients who underwent unilateral PCNL with single pole access were observed 1 day before and 5-7 days after operation with CDFI.Parameters were analyzed statistically.ResultsAfter operation,resistance index (RI) of renal arteries decreased (P<0.05).The diastolic flow statistically increased at main renal artery (MRA) of renal hilus,interlobar renal artery and interlobular renal artery (all P<0.05).After PCNL,in serious hydronephrosis patients,RI decreased (P<0.05) at segmental renal artery (SRA) and interlobar artery,end-diastolic flow velocity (Vmin) increased at interlobar renal artery (P<0.05).In moderate hydronephrosis patients,RI decreased at all renal arteries (P<0.05) after PCNL,Vmin increased at MRA and interlobular renal artery (P<0.05).In minor hydronephrosis patients,RI decreased at MRA and SRA,Vmin increased at SRA.In patients without hydronephrosis,RI changeed like serious hydronephrosis patients.ConclusionAfter PCNL,ipsilateral renal perfusion improves,renal diastolic flow increases and RI decreases.CDFI can be used to observe the blood perfusion of kidney,and provide quantitative information of renal hemodynamics.
3.Development of a CRISPR/Cpf1 gene editing system in silkworm Bombyx mori.
Zhanqi DONG ; Qi QIN ; Xinling ZHANG ; Kejie LI ; Peng CHEN ; Minhui PAN
Chinese Journal of Biotechnology 2021;37(12):4342-4350
The CRISPR/Cas9 gene editing system has been widely used in basic research, gene therapy and genetic engineering due to its high efficiency, fast speed and convenience. Meanwhile, the discovery of novel CRISPR/Cas systems in the microbial community also accelerated the emergence of novel gene editing tools. CRISPR/Cpf1 is the second type (V type) CRISPR system that can edit mammalian genome. Compared with the CRISPR/Cas9, CRISPR/Cpf1 can use 5'T-PAM rich region to increase the genome coverage, and has many advantages, such as sticky end of cleavage site and less homologous recombination repair. Here we constructed three CRISPR/Cpf1 (AsCpf1, FnCpf1 and LbCpf1) expression vectors in silkworm cells. We selected a highly conserved BmHSP60 gene and an ATPase family BmATAD3A gene to design the target gRNA, and constructed gHSP60-266 and gATAD3A-346 knockout vectors. The efficiency for editing the target genes BmATAD3A and BmHSP60 by AsCpf1, FnCpf1 and LbCpf1 were analyzed by T7E1 analysis and T-clone sequencing. Moreover, the effects of target gene knockout by different gene editing systems on the protein translation of BmHSP60 and BmATAD3A were analyzed by Western blotting. We demonstrate the CRISPR/Cpf1 gene editing system developed in this study could effectively edit the silkworm genome, thus providing a novel method for silkworm gene function research, genetic engineering and genetic breeding.
Animals
;
Bombyx/metabolism*
;
CRISPR-Cas Systems/genetics*
;
Endonucleases/genetics*
;
Gene Editing
;
RNA, Guide/genetics*
4.Mechanism analysis of Anti-BmNPV resistant strain NC99R.
Zhanqi DONG ; Xuejiao LEI ; Qi QIN ; Xinling ZHANG ; Liang TANG ; Meining SHI ; Minhui PAN
Chinese Journal of Biotechnology 2020;36(1):100-108
Bombyx mori is a lepidopteran insect with important economic value. Bombyx mori nucleopolyhedrovirus (BmNPV) causes huge economic loss to silkworm industry in China every year. The objective of this study is to determine the anti-BmNPV mechanism of Bombyx mori strain NC99R, and to provide a basis for understanding the molecular mechanism of the silkworm resistance strain. The normal control Dazao (DZ) strain and the NC99R resistant strain were fed with occlusion bodies (OB). The median lethal dose (LD50) analysis of the DZ and NC99R showed that the LD50 of DZ was 1.2×10⁵ OBs/larva, while NC99R was 1.8×10⁶ OBs/larva. The LD50 of the NC99R was about 15 times higher than the DZ. The mortality of DZ and NC99R were analyzed, which were fed with 1×10⁶ OBs/larva and injection with 1×10⁶ BVs/larva. The results showed that the death peak of DZ was concentrated in the 4th to 6th day. And the death peak of NC99R was concentrated in the 6th to 8th day, with a delay of 1-2 days compared with the control. The BmNPV DNA copy number showed that the BmNPV genome in DZ proliferated rapidly. The copy number of BmNPV DNA in NC99R were increased slowly after oral infection and body injection. HE staining showed that midgut tissue has no significant difference between DZ and NC99R in the early stage of oral infection. At 96 h p.i., the nucleus of DZ midgut became larger and shedding. The NC99R had enlarged nuclei, but the cells were still arranged neatly. Finally, the expression of virus genes in different periods were analyzed by RT-PCR. The results indicated that the immediate early gene ie-1 expression levels began to down-regulate after 24 h p.i.. The early, late, and extremely late genes were also down-regulated, and finally maintained at a lower expression level.
5.Leaky Gut Plays a Critical Role in the Pathophysiology of Autism in Mice by Activating the Lipopolysaccharide-Mediated Toll-Like Receptor 4-Myeloid Differentiation Factor 88-Nuclear Factor Kappa B Signaling Pathway.
Fang LI ; Haoran KE ; Siqi WANG ; Wei MAO ; Cexiong FU ; Xi CHEN ; Qingqing FU ; Xiaori QIN ; Yonghua HUANG ; Bidan LI ; Shibing LI ; Jingying XING ; Minhui WANG ; Wenlin DENG
Neuroscience Bulletin 2023;39(6):911-928
Increased intestinal barrier permeability, leaky gut, has been reported in patients with autism. However, its contribution to the development of autism has not been determined. We selected dextran sulfate sodium (DSS) to disrupt and metformin to repair the intestinal barrier in BTBR T+tf/J autistic mice to test this hypothesis. DSS treatment resulted in a decreased affinity for social proximity; however, autistic behaviors in mice were improved after the administration of metformin. We found an increased affinity for social proximity/social memory and decreased repetitive and anxiety-related behaviors. The concentration of lipopolysaccharides in blood decreased after the administration of metformin. The expression levels of the key molecules in the toll-like receptor 4 (TLR4)-myeloid differentiation factor 88 (MyD88)-nuclear factor kappa B (NF-κB) pathway and their downstream inflammatory cytokines in the cerebral cortex were both repressed. Thus, "leaky gut" could be a trigger for the development of autism via activation of the lipopolysaccharide-mediated TLR4-MyD88-NF-κB pathway.
Mice
;
Animals
;
NF-kappa B
;
Myeloid Differentiation Factor 88/metabolism*
;
Lipopolysaccharides/pharmacology*
;
Toll-Like Receptor 4/metabolism*
;
Autistic Disorder/metabolism*
;
Signal Transduction/physiology*