Cloning and expression of SmDXS2 gene in Swertia mussotii / 中国中药杂志

1-deoxy-D-xylulose-5-phosphate synthase2(DXS2) is the first key enzyme of the MEP pathway,which plays an important role in terpene biosynthesis of plants. According to the data of Swertia mussotii transcriptome, DXS2 gene(Gen Bank number MH535905) was cloned and named as Sm DXS2. The bioinformatics results showed that Sm DXS2 has no intron,with a 2 145 bp open reading frame encoding a polypeptide of 714 amino acids. They are belonging to 20 kinds of amino acids,and the most abundant amino acids include Ala,Gly and Trp. The predicted protein molecular weight was 76. 91 k Da and its theoretical isoelectric point(p I) was6. 5,which belonging to a hydrophilic protein. α-Helix and loop were the major motifs of predicted secondary structure of DXS2. The three function domains are TPP＿superfamily,Transket＿pyr＿ superfamily and Transketolase＿C superfamily,respectively. The Sm DXS2 protein shared high identity with other DXS2 proteins of plants. Phylogenetic analysis showed that Sm DXS2 protein is grouped with the gentian DXS2 protein. The recombinant protein of Sm DXS2 gene in Escherichia coli was approximately 92. 00 k Da(containing sumo-His tag protein 13 k Da),which was consistent with the anticipated size.This work will provide a foundation for further functional research of Sm DXS2 protein and increasing the product of iridoid compound by genetic engineering in S. mussotii.

Increased autoantibody production against AT(1)-receptors and alpha(1)-adrenergic receptors in hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe autoantibodies production against AT(1)-receptors and alpha(1)-adrenergic receptors and association to risk factors, such as sex, age, family history, course of hypertension and other cardiovascular diseases in hypertensive patients.</p><p><b>METHODS</b>A total of 690 patients with essential hypertension admitted to our hospital were selected and autoantibodies against AT(1)-receptors and alpha(1)-adrenergic receptors were detected by ELISA. Multiple logistic regression analysis was performed based on obtained data.</p><p><b>RESULTS</b>Positive rates for antibody against AT(1)-receptors and alpha(1)-adrenergic receptors were 47.1% (325/690) and 36.4% (251/690) respectively in this group of patients. Duration of hypertension history was significantly longer in the antibody against AT(1)-receptors and alpha(1)-adrenergic receptors positive groups [(9.3 +/- 11.0) year, (9.9 +/- 11.1) year] compared to the negative groups [(7.3 +/- 9.3) year, (7.2 +/- 9.5) year, all P < 0.01]. The ratio of family history with hypertension was also significantly higher in antibody positive groups than negative ones (47.69% vs 39.18%, P < 0.01). Regression analysis demonstrated that 5 risk factors were related to positive production of autoantibody against AT(1)-receptors including female gender, age, family history, duration of hypertension history and diabetes. However, just age, family history, duration of hypertension history were main factors responsible to the production of autoantibody against alpha(1)-adrenergic receptors (all P < 0.05).</p><p><b>CONCLUSION</b>The environmental and genetic factors contributed to the autoantibody production in patients with essential hypertension.</p>

Association between ADRA1A gene polymorphism and autoantibodies against the alpha1-adrenergic receptor in hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the association between ADRA1A gene polymorphism and autoantibodies against the alpha1-adrenergic receptor in hypertensive patients.</p><p><b>METHODS</b>A total of 396 patients with essential hypertension admitted to our hospital were selected and autoantibodies in sera were detected by ELISA, and patients were divided into the autoantibody positive and negative group. Genomic DNA was extracted from erythrocytes obtained from EDTA-treated blood by the Blood DNA extraction kit. Gene polymorphisms were detected by ligase detection reaction (LDR), including rs574584, rs1048101, rs3739216 and rs3802241. The frequency of genotypes and haplotype were analyzed.</p><p><b>RESULTS</b>The frequencies of detected genotypes between the autoantibody against the alpha1-adrenergic receptor positive group and negative group were similar (P > 0.05) while significant difference was in the frequencies of haplotypes (all P < 0.05). The frequencies of genotypes with rs1048101 (genotype C/C, C/T, P = 0.017) and rs3802241 (genotype A/A, A/G, P = 0.004) were significant different in autoantibody positive group compared to negative group in patients with stage 2.</p><p><b>CONCLUSION</b>ADRA1A gene polymorphism might correlate with the alpha1-adrenergic receptor autoantibody production in hypertensive patients.</p>

The mechanism of signal transduction during vascular smooth muscle cell proliferation induced by autoantibodies against angiotensin AT1 receptor from hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Autoantibodies against angiotensin AT1 receptor have been discovered in patients with preeclampsia or malignant hypertension. Some studies have demonstrated that the autoantibodies are involved in the immunopathogenesis of hypertension and have an agonist effect similar to angiotensin II.</p><p><b>METHODS</b>Autoantibodies against AT1 receptor were purified from sera of patients with primary hypertension by affinity chromatography. Proliferation of cultured rat vascular smooth muscle cells was detected by bromodeoxyuridine incorporation and activation of signalling molecules detected by Western blotting and electrophoretic mobility shift assay.</p><p><b>RESULTS</b>The AT1-RAb caused a significant proliferation similar to the Ang II during first 24 hours. The levels of nuclear factor-kappaB (NF-kappaB), phosphorylated JAK2, phosphorylated STAT1 (pSTAT1) and phosphorylated STAT3 (pSTAT3) molecules were increased in response to the autoantibodies. In contrast, the activations of NF-kappaB and JAK-STAT were blocked by losartan, pyrrolidinedithiocarbamate (a specific inhibitor of NF-kappaB) and AG490 (a specific inhibitor of the JAK2 tyrosine kinase). The expressions of NF-kappaB, pSTAT1 and pSTAT3 reached peak levels at different times. Moreover, the relative densities of electrophoretic bands showed that activation of pSTAT3 was more significant than STAT1 induced by AT1-RAb.</p><p><b>CONCLUSIONS</b>These results suggest that the autoantibodies against AT1 receptor have an agonist effect similar to Ang II in proliferation of VSMCs and the NF-kappaB and JAK-STAT proteins play essential roles. The effect is different from Ang II in that STAT3 is the main downstream activating molecule in JAK-STAT signalling pathway.</p>

Autoantibodies against alpha1 adrenergic receptor related with cardiac remodeling in hypertensive patients by clinical observation / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of autoantibodies against alpha(-) adrenergic receptor on cardiac remodeling in patients with hypertension.</p><p><b>METHODS</b>Five hundred and fifty three patients with hypertension in our hospital were selected. The autoantibodies against alpha(1) adrenergic receptor in sera of donor were detected by ELISA, and the results of echocardiography were recorded. By multiple logistic regressions, the risk factors were analyzed on left ventricular enlargement of hypertension.</p><p><b>RESULTS</b>The percentage of autoantibodies against alpha(1) adrenergic receptor positive was 32.3% (179/553). There were significant difference between the positive group and negative group on the ratio of left atrial enlargement (53.6%, 44.3%, respectively; P < 0.05) and left ventricular enlargement (12.8%, 6.1%, respectively; P < 0.01). The result of regression analysis demonstrated that 4 risk factors were related to left ventricular enlargement, including male, course of disease, heart rate (HR) and autoantibodies against alpha(1) adrenergic receptor in the serum (all P < 0.05).</p><p><b>CONCLUSIONS</b>The autoantibodies against alpha(1) adrenergic receptor have a relationship with left ventricular enlargement of hypertension. Patients with the activity of autoantibodies against alpha(1) adrenergic might contribute to predict cardiac remodeling.</p>

VSMCs Proliferation and Mechanism of Signal Transduction by Angiotensin Ⅱ Type 1 Receptor Autoantibodies Mediated from Hypertensive Patients / 中华高血压杂志

Objective The autoantibodies against angiotensin Ⅱ type 1 receptor(AT_1 RAb)have been dis- covered in the patients with malignant hypertensive and preeclampsia,this autoantiboies(AT_1-AA)have an ago- nist-like activity effect similar to angiotensin Ⅱ(Ang Ⅱ).This study aimed at investigation the effect of Ang Ⅱ agonist-like activity by AT_1-AA on VSMCs proliferation was obtained from essential hypertensive patients. Methods VSMCs were cultured from aorta of WKY rats.The hypertensive patients" serum was purified by am- monium sulfate precipitation and affinity chromatography.The effect on VSMC proliferation this autoantilody was determined by BrdU incorporation.Total protein and the expression of phosphorylation JAK-STAT were assessed by Western blotting.Results AT_1RAb caused a significant increase in BrdU incorporation similar to Ang Ⅱ during 0-24 h reaching peak value at 12 h.The A value of in 450 nm was higher in AT_1RAb group (0.236?0.012)than AG490+AT_1RAb group(0.176?0.009),Losartan+AT_1RAb groups(0.119?0.006) and Serum Free group(0.127?0.006)(P

A preliminary investigation on the serological and epidemiological characteristics of severe acute respiratory syndrome in children / 中华儿科杂志

<p><b>OBJECTIVE</b>The severe acute respiratory syndrome (SARS) is a highly contagious infection caused by a newly discovered strain of coronavirus (SARS-CoV). During the outbreak of SARS in the first half of 2003, children appeared to be less susceptible to the SARS coronavirus and pediatric patients presented with a less aggressive clinical course than adult patients did, demonstrating the traits which were rarely observed in other viral contagious disease. The present study aimed to preliminarily examine the presence of serum specific antibodies against severe acute respiratory syndrome (SARS)-associated coronavirus virus (SARS-CoV) in pediatric SARS patients and explore the possibility of subclinical infection in children/adults through close association with SARS cases.</p><p><b>METHODS</b>(1) Clinicians and nurses visited families and collected general and epidemiological information about the subjects using a standard questionnaire and took serum specimens. (2) Specific antibodies against SARS-CoV were assayed with two methods, indirect immunofluorescence assay (IFA) for detecting IgG antibodies and enzyme linked immunosorbent assay (ELISA) for mixed antibodies. Serum specimens tested included those from 21 clinically confirmed pediatric SARS cases (aged from 8 months to 14 years, 11 male and 10 female) and their 23 parents who had close contact with the children, 36 adult patients in convalescence stage of SARS, 24 children (aged 1.5 to 14 years) and other 34 adults who had close contact with infected adults.</p><p><b>RESULTS</b>(1) The positive rates of specific IgG and mixed antibodies against SARS-CoV were 38% (8/21) and 33% (7/21) in pediatric cases; whereas the rates were 75% (27/36) and 69% (25/36) in adult patients. (2) The proportion of the patients who had close contact to SARS patients was 7/8 among the antibody-positive group vs. 1/13 for the antibody-negative group (P < 0.05). (3) The IgG antibody emerged in one of 24 children, whose mother, a nurse, had suffered from SARS (4%). (4) Among 23 parents of children with SARS, one was positive for IgG and the mixed antibodies, whose grandson and husband suffered from SARS; The IgG antibody and the mixed antibodies were also positive in another adult who had close contact with adult SARS cases (3%).</p><p><b>CONCLUSIONS</b>(1) SARS-CoV infection was confirmed by serological methods in 38.1% of clinically diagnosed pediatric SARS cases, which leads to the assumption that correct diagnosis of pediatric SARS requires more accurate and efficient ways, for example, screening for antigen or gene of SARS-CoV. (2) The proportion of the patients who had close contact to SARS patients among antibody-positive cases was higher than that in antibody-negative cases. (3) It is possible that subclinical SARS CoV infection exists in children and adults, although the rate of occurrence is low. The data of the present study did not confirm that SARS had subclinical infection among adults who had close contact to pediatric SARS cases.</p>