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Objectiveezrin gene is overexpressed in pancreatic cancer, and its upstream sequence plays an important role in gene expression. This study intends to knock out ezrin transcriptional regulatory region and identify its gRNA target sites for gene editing in pancreatic cancer cells.MethodsThe reporter gene expression vectors carrying the upstream segment of ezrin gene were transiently transfected into Panc-1 cells. The ezrin transcriptional regulatory regions were identified by double luciferase reporter gene detection system. Then, the online software was utilized to predict the gRNA target sites located at the upstream and downstream of ezrin transcriptional regulatory region. Two recombinant plasmids pX459-sgRNA-L and pX458-sgRNA-R contained these two sequences were constructed for gene editing. Moreover, in order to identify the targeted knockout of ezrin transcriptional regulatory region, the recombinant plasmids were co-transfected into Panc-1 cells, and the genome DNA contained gRNA target sites were amplified, subcloned and sequenced. Finally, Panc-1 cells transfected with recombinant plasmids were preliminary sorted using puromycin treatment. The cell proliferation was detected by water-soluble tetrazolium salt method.ResultsLuciferase data showed that ezrin gene fragment -1297/-1186 enhanced the transcriptional activity of SV40 promoter and ezrin promoter in Panc-1 cells. Subclonal sequencing data revealed that the recombinant plasmids carrying the gRNA target sequence of ezrin transcriptional regulatory region were co-transfected into Panc-1 cells could trigger the genomic DNA fragments, which located between gRNA-L and gRNA-R target sites. Cell proliferation assay showed that the proliferation was significantly inhibited after transfection.ConclusionThe targeted knockout of ezrin transcriptional regulatory region was achieved and the inhibition of Panc-1 cell proliferation may be related to this knockout.
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Objective To investigate the effectiveness and safety of minimal invasive surgery combined with ventriculoscope for the treatment of basal ganglia hemorrhage.Methods From June 2014 to June 2015 there were 57 patients with basal ganglia hemorrhage were selected into this study.These patients were signed into the minimal invasive surgery combined with ventriculoscope group and the craniotomy group according to the methods of surgery they received.And the clinical outcomes of the two groups were compared.Results Compared with the craniotomy group,patients in the minimal invasive surgery combined with ventriculoscope group got a significantly lower Glasgow coma scale at 7 days,14 days and 28 days after the operation (P =0.02,0.04,0.04);the hospital stays were significantly reduced in the minimal invasive surgery combined with ventriculoscope group [(21.45 ±5.67)d vs.(25.67 ±7.45)d,P =0.02];and the operation time were significantly reduced as well [(134.45 ±21.11)min vs.(178.65 ±45.32)min,P =0.000)].There was no significant difference in intra-cranial pressure,rate of hematoma clearance,rate of organ functional failure,rate of re-bleeding and mortality 28 days after operation (P >0.05).Conclusion The minimal invasive surgery combined with ventriculoscope is effective and safe for the treatment of basal ganglia hemorrhage,which is worthy of popularization.
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<p><b>BACKGROUND</b>Data on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.</p><p><b>METHODS</b>The survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.</p><p><b>RESULTS</b>The analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).</p><p><b>CONCLUSIONS</b>The prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Awareness , Hypertension , Epidemiology , Therapeutics , Prevalence , Renal Insufficiency, ChronicABSTRACT
OBJECTIVE: To design and synthesize a series of novel amino acid-modified ursolic acid derivatives containing primary amino-group and to investigate their antitumor activities against gastric carcinama in vitro. METHODS: 1, 2-Ethylenediamine was added to the C28 site of ursolic acid(UA) and the resulting intermediates were conjugated to amino acid to get the amino acid-modified ursolic acid derivatives. The effects of these UA derivatives on the growth of BGC823 cells and AGS cells were assessed by MTT assay. Meanwhile, Annexin V/PI dual staining and cell cycle analysis were performed to investigate the anti-tumor mechanism. RESULTS: Most of the derivatives exhibited more powerful cytotoxicity than UA against these two gastric cancer cell lines. The apoptosis of BGC823 cells incubated with compound 3 or compound 4 were observed. Cell cycle analysis showed that compound 3-6 triggered 44.69% -92.64% of the treated AGS cells into apoptotic status. CONCLUSION: Potent anticancer abilities of these UA derivatives bearing primary amido are achieved by inducing apoptosis. Introducing primary amino group to UA is an effective way to develop new UA derivatives with enhanced anti-proliferative activities. Copyright 2012 by the Chinese Pharmaceutical Association.
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<p><b>OBJECTIVE</b>To identify the genetic defects in patients with congenital atrial septal defects (ASD).</p><p><b>METHODS</b>The clinical data and blood samples from 180 unrelated subjects with congenital ASD were collected and evaluated. Two hundred healthy individuals served as controls. The coding exons and the flanking introns of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with the sequences publicized in GenBank by the aid of programme BLAST to identify the sequence variations. Clustal W software was applied for analysis of the conservation of altered amino acids.</p><p><b>RESULTS</b>Two novel heterozygous missense GATA4 mutations were identified in 2 out of 180 ASD patients. Namely, the triplet substitutions of GTC for GGC at codon 21 and TCG for CCG at codon 87 were detected, predicting the conversions of glycine into valine at amino acid residue 21 (G21V) and proline into serine at amino acid residue 87 (P87S). None of the two mutations were detected in 200 healthy controls. Across-species alignment of GATA4 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, a single nucleotide polymorphism c.99G>T was observed. However, the polymorphic frequency distribution in ASD cases was similar with that in healthy controls (for genotype GT, χ(2) = 0.7556, P = 0.3847; for allele T, χ(2) = 0.7235, P = 0.3950).</p><p><b>CONCLUSIONS</b>Two novel mutations of GATA4 gene are identified in two unrelated ASD patients. This finding provides new insight into the molecular etiology responsible for ASD.</p>