Transforming growth factor-?_1 (TGF-?_) mRNA or protein expression in renal cortex of diabetic rats was assessed by real-time quantitative RT-PCR with SYBR Green,immunohistochemistry or Western blot.After melatonin treatment,the expressions of TGF-?_1 mRNA and protein were decreased,suggesting that beneficial effect of melatonin may result from its antioxidative property and inhibiting TGF-?_1 expressions.

Glucose-6-phosphate dehydrogenase is main regulatory enzyme for pentose phosphate pathway. To amplify the core sequence of G6PDH gene from Chimonanthus praecox, the primers were synthesized, based on the conserved nucleotide sequence of other reported plant G6PDH genes. The specific primers were designed according to the major fragment. The full length cDNA of the G6PDH1 gene was isolated by the 3' and 5' rapid amplification of cDNA ends approach. Transcript levels of G6PDH1 isoform was measured by real-time quantitative RT-PCR in different tissues and in responds to cold treatment. The G6PDH1 subcellular localization, transmembrane domain, three-dimensional structure, and phylogenetic analysis were predicted by different software to analysis the bioinformatics of G6PDH1 protein. The G6PDH1 cDNA sequence was 2 011 bp in length and consisted of 1 551 bp Open Reading Frame (ORF) , encoding a protein of 516 amino acids. Expression analysis results in different tissues showed that G6PDH1 was primarily observed in flowers and roots, as opposed to the leaves and stems. Cold treatment experiments indicated that cold treatment caused a rapid increase in G6PDH1 expression in flowers within 12 h. The full-length cDNA of G6PDH1 and its expression analysis will play an important role for further study on cold stress responses in Ch. praecox.
Calycanthaceae ; chemistry ; classification ; enzymology ; genetics ; Cloning, Molecular ; Enzyme Stability ; Glucosephosphate Dehydrogenase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism

This study is to establish an HPLC-DAD-ELSD method for simultaneous determination of 5 flavones and saponins in Rhizoma Anemarrhenae including neo-mangiferin, mangiferin, timosaponin B II, timosaponin B III and timosaponin A III. Samples were analyzed on a Merck Purospher STAR column(4.6 mm x 250 mm, 5 μm). The mobile phase consisted of acetonitrile( A) and 0. 1% formic acid (B) with gradient elution at a flow rate of 1.0 mL · min(-1). The column temperature was set at 40 °C. The DAD detector wavelength was set at 254 nm. The ELSD conditions were as follows: the nebulizing gas flow rate was 2.0 L · min(-1) and temperature of drift tube was 105 °C. The volume was 10 μL. The five compounds were well separated with good linear correlations. The mean recoveries were between 102.0%-104.0%. This method was quick and reliable which provides a foundation for quality control of R. Anemarrhenae.
Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Rhizome ; chemistry ; Saponins ; analysis

Objective To evaluate the effect of the Parasep? feces centrifuge tube method on detecting schistosome eggs. Methods A total of 803 residents aged from 6-65 years were selected in 2 schistosomiasis endemic villages Jiangling Coun-ty Hubei Province and their stool samples were collected and detected parallelly by the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method at the same time. Results Among the 803 people 15 cases were found of schistosome egg positive and the positive rate was 1.87%. The positive rates of the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method were 0.75% 1.49% and 1.12% respectively. The schistosome eggs got with the Parasep? feces centrifuge tube method were clear and easy to identify. Conclusion In low endemic areas of schistosomiasis the Parasep? feces centrifuge tube method can be used as schistosomiasis japonica etiology diagnosis method.

OBJECTIVESTo investigate targeted blockage of BCR/ABL oncoprotein mediated cell transformation by STAT5 decoy oligodeoxynucleotide (ODN), its effect on the growth and proliferation inhibition of K562 cells and the related molecular mechanisms.

METHODSSTAT5 decoy ODN, designed and synthesized in vitro, was transfected into K562 cells by cationic lipid. The cell growth curve and colony formation assay were used to reflect the growth and proliferation capacity of K562 cells, RT-PCR to detect the expression of three genes downstream STAT5.

RESULTSConfocal microscopy demonstrated that STAT5 decoy ODN was successfully transfected into K562 cells (95.2% positive cells). STAT5 decoy ODN inhibited the growth of K562 cells (inhibition rate 77.7%) and their colony formation capacity (Decoy ODN treated group 8.3% vs control group 35.7%, P < 0.05) after the treatment with STAT5 decoy ODN, the expressions of c-myc, bcl-X(L), cyclin D1 mRNA were down-regulated by 15.4%, 30.8%, 29.1%, respectively in the K562 cells.

CONCLUSIONSSTAT5 decoy ODN inhibits the growth and proliferation of K562 cells. The mechanisms may be that decoy ODN blocks the transcriptional activation potent of STAT5 and down-regulates the expression of these tumor related genes downstream STAT5.

Cell Proliferation ; Cyclin D1 ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; Microscopy, Confocal ; Oligodeoxyribonucleotides, Antisense ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; STAT5 Transcription Factor ; genetics ; physiology ; Transfection ; bcl-X Protein ; genetics

To clarify the necessity of applying appropriate statistical methods to calculate sampling error from data of cluster sample survey, we take a two-stage cluster sample survey developed from a population aged 15 and over as example. We use statistical methods based on the assumption of simple random samples and methods considering complex sample design to analyze our data, respectively. Through comparison, we hope to show the potential effects of using improper statistical methods to estimate sampling error on parameter estimation and hypothesis testing. Using standard error algorithms based on the assumption of simple random samples,the standard errors calculated often underestimate sampling error and the hypothesis testing even gets wrong conclusion. When the statistical methods and statistics package for complex survey data are already available, it is necessary for us to use appropriate methods to analyze and report the sampling errors of data from cluster sample survey.
Cluster Analysis ; Data Interpretation, Statistical ; Epidemiologic Studies ; Sampling Studies

OBJECTIVETo study the quantitative and functional changes of peripheral blood dendritic cells (DCs) and their subsets in the leukocyte population in patients with coronary artery disease (CHD) with different coronary artery plaques and explore the relation between DCs and coronary plaque development.

METHODSThirty CHD patients were divided into SAP (10 cases), UAP (10 cases) and ACS (10 cases) groups, with another 10 patients having negative result in coronary angiography as the control group. Intravascular ultrasound (IVUS) was performed to identify the nature of the plaques. The percentage and absolute number of peripheral blood DCs and DC subsets were measured by flow cytometry. The functional status of the DCs was analyzed by enzyme-linked immunosorbent assay (ELISA) and flow cytometry.

RESULTSIn the SAP group, IVUS found stable plaques in 8 cases and unstable plaques in 2 cases; in UAP group, 7 patients had unstable plaques, 2 had stable plaques, and 1 had plaque rupture. Plaque rupture, unstable plaques and stable plaques were found in 6, 3 and 1 patients in ACS group, respectively. In comparison with patients with stable plaques, those with unstable plaques had significantly increased percentages and number of DCs, mDCs and mDC1 (P<0.05), while the mDC2s and pDCs showed no obvious difference between them (P>0.05). The percentages and number of DCs, mDCs, mDC1s and pDCs were significantly decreased in patients with ruptured plaques (P<0.05). In peripheral blood monouclear cells cultured for 7 days, the CD83 expression was significantly higher in unstable and rupture plaque groups than in stable plaque group, and no significant difference was found between stable plaque group and the control group (P>0.05). In unstable and rupture plaque groups, co-culture with 2x10(5)/ml DCs evoked strong proliferation of the T cells in comparison with the stable plaque group, but no difference was found between the stable plaque and the control groups (P>0.05). Significantly higher levels of interleukin-2 and interferon-alpha were detected in the supernatant of the mixed lymphocyte reaction in unstable and ruptured plaque groups than in stable plaque and control groups, without obvious difference between the latter two groups.

CONCLUSIONThe percentage and absolute number of peripheral blood DCs and their functional status suggest the alterations of the coronary artery plaques in CHD patients.

Case-Control Studies ; Cells, Cultured ; Coronary Angiography ; Coronary Artery Disease ; immunology ; pathology ; Coronary Vessels ; pathology ; Dendritic Cells ; classification ; cytology ; immunology ; Female ; Flow Cytometry ; Humans ; Male

OBJECTIVETo investigate the effect of telomerase reverse transcriptase (TERT) to the proliferation of 5-HT induced pulmonary artery smooth muscle cells (PASMCs).

METHODSThe PASMCs proliferation experiment was performed to detect the effort on PASMCs of 5-HT or ASODN TERT (antisense oligoribonucleotides TERT designed according to the rat TERT mRNA sequence of gene bank). The immunohistochemistry staining experiment and the in situ hybridization experiment were to detect the TERT protein and mRNA expression with 5-HT or ASODN TERT. FITC marked ASODN TERT experiment was done to research the distribution of ASODN TERT in PASMCs.

RESULTS5-HT promoted PASMCs proliferation in a dose-dependent manner (10(-9) - 10(-5) mol/L). 5-HT also significantly increased TERT expression at protein and mRNA levels as shown by immunohistochemistry staining and the in situ hybridization studies. This effect could be blocked by ASODN TERT in a time and dose-dependent manner.

CONCLUSIONSOur experiments show TERT is one of the key factors in the procession of 5-HT induced PASMCs proliferation. ASODN TERT might be a potential therapy agent for pulmonary hypertension.

Animals ; Cell Proliferation ; Cells, Cultured ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Pulmonary Artery ; cytology ; RNA Replicase ; RNA, Messenger ; genetics ; Rats ; Serotonin ; pharmacology ; Telomerase ; pharmacology

In order to evaluate the efficiency of ITS2 and psbA-trnH sequences used as DNA barcodes to distinguish Plantaginis Semen from its adulterants, we collected 71 samples of Plantaginis Semen and its adulterants. The ITS2 and psbA-trnH sequences were aligned through Clustal W, and the genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining (NJ) phylogenetic trees were constructed using MEGA 5.1. The results indicated that the ITS2 sequence lengths of Plantago asiatica and P. depressa were 199 bp and 200 bp, respectively; the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance; the NJ tree based on ITS2 sequence indicated that Plantaginis Semen and its adulterants could be distinguished clearly. The sequence lengths of psbA-trnH of both P. asiatica and P. depressa were 340 bp; the maximum intra-specific K2P distances were lower than the minimum inter-specific K2P distance; the NJ tree based on psbA-trnH sequence showed that Plantaginis Semen can be distinguished clearly from its adulterants except for P. major. Therefore, ITS2 sequences can be used as an ideal DNA barcode to distinguish Plantaginis Semen from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plantago ; classification ; genetics ; Quality Control ; Seeds ; classification ; genetics

Objective Familial left ventricular noncompaction( LVNC ) is quite rare.We screened for the presence of LVNC and related clinical characteristics in a 5-generation Chinese family.Methods Comprehensive medical history was obtained from 40 members in a 5-generation Chinese family.Systemic clinical investigations including echocardiography (UCG),routine and ambulatory electrocardiogram (ECG),X-rays were performed in 33 family members.Cardiovascular magnetic resonance image (MRI) was carried out in 2 family members.Results Sudden cardiac death (including 1 occurred while following-up)was reported in 7 family mcmbers( 17.5％,7/40 ).LVNC was diagnosed in 10 out of the 33 family members (30.3％ )and heart enlargement was evidenced in 3,heart failure in 2,complete left branch conductive block in 3,serious sick sinus syndrome (SSS) treated with permanent pacemaker implantation in 1 and paroxysmal supraventricular tachycardia treated with radiofrequency ablation procedure in 1 out of these 10 LVNC patients.Primary pedigree analysis revealed that offspring from female patients were at the highest risk to be affected by LVNC ( 15/18,83.3％ ) while LVNC was absent in offspring of male LVNC patients (0/8).Moreover,clinical heart failure symptoms and arrhythmias were more severe in female LVNC patients than in male LVNC patients.Conclusion Primary familial investigation reveals the matrilineal inheritauce of familial LVNC in this 5-generation Chinese family,further investigations are warranted to explore the potential mutations in the mitochondrial genome responsible for LVNC in this family.