Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease ( TALEN) mRNAs.Methods TALEN knockout site recognizing Txnip was designed by tools on line, then constructed the vectors and assayed its cleavage activity at cellular level.TALEN mRNA was transcribed in vitro and microinjected into C57BL/6J mouse zygotes.F0 mice were verified at DNA level with BamHI and TXNIP-knockout mice were obtained.Results We designed and constructed TALENs which recognized and cut the first exon of Txnip, and got four TXNIP knockout mice, among which two were frameshift mutation, demonstrating that the TXNIP-knockout mice were generated by TALEN technique.Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

Parkinson's disease (PD) is a neurodegenerative movement disorder mainly due to degeneration of dopaminergic neurons in the substantia nigra. Most PD cases are sporadic and only 5%-10% of patients carry mutations with inheritance. Among them, the mutation of DJ-1 is related to the autosomal recessive early-onset parkinsonism. DJ-1, the Parkinson's disease-related protein, plays important roles in different physiopathological processes, including oxidative stress, cell translocation and regulation of transcription and translation. DJ-1 is known to be widely expressed in different areas of brain, including hippocampus, amygdala, substantia nigra and cortical areas. Several researchers have tried to demonstrate the clinical and neuroimaging features of DJ-1 related parkinsonism. The DJ-1 knockout mouse model was established to further explore the mechanisms of different functions. Moreover, the search for different forms of DJ-1 as potential biomarkers of PD also provides guidance for its accurate diagnosis and treatment in the future.
Animals ; Dopaminergic Neurons ; Mice ; Oncogene Proteins ; Oxidative Stress ; Parkinson Disease ; Protein Deglycase DJ-1 ; Substantia Nigra

Background: Although the association of single nucleotide polymorphisms (SNPs) of cyclooxygenase (COX) genes and the risk of aspirin resistance (AR) has been extensively studied, the results remain conflicting. The majority of studies have focused on the role of rs20417 (COX-2 -G765C) in AR. To derive a more comprehensive and accurate evaluation of this association, we performed a meta-analysis including the most recent studies. Methods: Relevant studies published up to October 2018 were identified by searching the PubMed, EMBASE, Web of Science, Cochrane, China Nation Knowledge Infrastructure Platform, Wanfang, and VIP databases, and by manual searching reference lists of the retrieved articles. Odds ratios (ORs) and 95% confidence intervals (CIs) were applied to assess the strength of associations. Sensitivity and subgroup analyses were performed to explore the stability of results and between-study heterogeneity, respectively. Results: A total of 18 studies on rs20417 were pooled into the meta-analysis. Rs20417 was found to be associated with an increased risk of AR (C vs. G: OR = 1.43, 95% CI = 1.10–1.86, p < 0.05; GC+CC vs. GG: OR = 1.54, 95% CI =1.15–2.05, p < 0.05). These associations were stronger in Chinese participants and in patients with ischemic stroke in subgroup analyses. Conclusion: The presence of rs20417 indicates an increased risk of AR, especially in Chinese participants and patients with ischemic stroke. This association could help to improve personalized medicine and initiate appropriate treatment as necessary. Further large-scale studies are warranted to confirm our findings.

Objective:To construct 68Ga-1, 4, 7-trizacyclononane-1, 4, 7-triacetic acid (NOTA)-CD44 as a novel atherosclerosis tracer targeting hyaluronic acid (HA), and evaluate its biological property and molecular imaging features. Methods:Low molecular weight (LMW) recombinant human CD44 protein was selected, and the C-terminal of the protein was modified by sulfonation and coupled to the bifunctional ligand NOTA to synthesize a novel molecular probe 68Ga-NOTA-CD44 targeting HA. The biological properties of the probe, such as labeling rate and in vitro stability, were studied. Three atherosclerotic plaque model mice and three normal C57BL/6 mice were studied by 68Ga-NOTA-CD44 microPET/CT imaging and pathological examination. Results:68Ga-NOTA-CD44 tracer was synthesized and purified with the radiochemical purity above 99%, and the specific activity was up to 62.22 MBq/nmol. lts stability was good in PBS, and the radiochemical purity was over 90% after incubation for 3 h. After intravenous injection, the probe was metabolized mainly by the kidneys, and its metabolic level decreased successively in the liver, lungs and blood. MicroPET/CT imaging results of atherosclerotic model mice suggested that the uptake in the plaque of abdominal aorta was higher at 60 min after injection, with SUV max and target/background ratio (TBR) max of 1.14±0.02 and 4.95±0.93, and the probe had certain atherosclerotic plaque eroded targeting, which was consistent with the pathological result. Conclusions:As a novel probe, 68Ga-NOTA-CD44 is simple to prepare and has a high labeling rate. It has good physicochemical properties and in vivo biological properties, and can display atherosclerotic eroded plaques sensitively. 68Ga-NOTA-CD44 has a promising prospect to be a new molecular probe for early noninvasive recognition of atherosclerotic eroded plaques.