Identification of ezrin gene transcriptional regulatory region and its gRNA target site in pancreatic cancer cells / 医学研究生学报

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.

Minimal invasive surgery combined with ventriculoscope for the treatment of basal ganglia hemorrhage / 局解手术学杂志

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.

Prevalence, awareness, treatment, and control of hypertension in the non-dialysis chronic kidney disease patients / 中华医学杂志(英文版)

<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>

Synthesis of amino acid-modified ursolic acid derivatives and their antitumor activity against gastric carcinoma / 中国药学杂志

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.

Novel GATA4 mutations identified in patients with congenital atrial septal defects / 中华心血管病杂志

<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>

Two new phthalides from Ligusticum chuanxiong.

Two new phthalide dimers, namely chuanxiongnolide A (1) and chuanxiongnolide B (2), along with eleven known phthalides and derivatives ( 3 - 13), were isolated from the roots of Ligusticum chuanxiong Hort. (Umbelliferae). The chemical structures of the new compounds were established by HR-ESI-MS, 1D- and 2D-NMR spectroscopic analysis, whilst the relative stereochemistry of 1 was confirmed by X-ray crystallographic data.

Three new diterpene alkaloids from the roots of Aconitum nagarum var. lasiandrum.

Three new diterpene alkaloids, 16,17-dihydro-12 beta,16 beta-epoxynapelline (1), N-deethyl- N-methyl-12-epi-napelline (2) and 11- epi-16 alpha,17-dihydroxylepenine (3), along with twenty-six known alkaloids (4 - 29) were isolated from the roots of Aconitum nagarum var. lasiandrum. The chemical structures of the new compounds were established by HR-MS as well as 1D- and 2D-NMR spectroscopic analysis. The absolute stereochemistry of 1 was confirmed by X-ray crystallography.

Hydrogen bonding and pi-pi stacking in dimethylgenistein.

The title compound, 5-hydroxy-4',7-dimethoxyisoflavone, C17H14O5, is composed of a benzopyranone moiety, a phenyl moiety and two methoxy groups. The benzopyranone ring is not coplanar with the phenyl ring, the dihedral angle between them being 56.28 (3) degrees. The two methoxy groups are nearly coplanar with their corresponding rings, having C-C-O-C torsion angles of 2.9 (2) and 5.9 (2) degrees . The molecules are linked by C-H...O hydrogen bonds into sheets containing classical centrosymmetric R2(2) (8) rings. The sheets are further linked by aromatic pi-pi stacking interactions and C-H...O hydrogen bonds into a supramolecular structure.

In vivo osteogenic properties of the composite of periosteal-derived osteoblast and collagen- coated true bone ceramics.

OBJECTIVE: To contrive new ways for repairing bone defects on the basis of the observation of the osteogenic properties of implanted composite made from periosteal-derived osteoblasts (POB) and true bone ceramics (TBC) coated with collagen (TBCc). METHODS: Bovine cancellous bone was calcined and coated with collagen before being mixed with cultured rabbit osteoblast suspension to prepare the composite, which was respectively implanted subcutaneously on the right of the back of BALB/c nude mice and in the muscle pouches in similar sites of rabbits. The control was established by implantation of TBCc in the contralateral sites corresponding to the former implants in both of animals. Samples were obtained from the implant sites 4 weeks later in mice and 8 weeks later in rabbits respectively, and light and electron microscopic observation was performed routinely. RESULTS: A large number of osteoblasts were seen to grow and proliferate within the TBCc-POB composite where cartilage and bone tissues also generated, which was no observed in the control group. CONCLUSION: The osteoblasts in the implanted TBCc-POB composite can survive and develop into cartilage and bone tissues.