The viral RNA was extracted from purified cymbidium mosa ic virus (CyMV) isolated from Dendrobium orchid cultivated in Hainan island. The gene of the movement protein (MP) was amplified by means of reverse transcripti on-polymerase chain reaction (RT-PCR), and cloned into pGEM-T easy vector. Se quence analysis showed that the gene fragment contained 3 open reading frames (O RFs) which may be encoding 14 kD、12 kD and 10 kD peptides. The nucleotide seque nce of the cloned gene fragment shared 97.8% homology with the MP genes of CyMV isolated from orchids cultivated in Hawaii and Singapore.

In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum
Acetonitriles ; Chromatography, High Pressure Liquid ; Flavonoids ; chemistry ; Kaempferols ; Quercetin ; analogs & derivatives ; Rutin ; Tandem Mass Spectrometry ; Vitaceae ; chemistry

The aim of the present study is to investigate the role of β(2)-adrenoreceptor (β(2)-AR) in ischemic preconditioning (IP) in isolated rat heart model of ischemia/reperfusion (I/R). Sprague-Dawley rat hearts were quickly removed, mounted on Langendorff apparatus, and perfused with Krebs-Henseleit (KH) solution. After the initial stabilization period, the rats were randomly divided into 6 groups including control group (perfused for an additional 20 min), IP group (4 cycles of 5 min of ischemia followed by 5 min of reflow), isoproterenol (ISO) group (10 nmol/L ISO perfusion for 5 min followed by 5 min washout), IP + ICI118551 group (55 nmol/L ICI118551 perfusion for 5 min before and throughout IP), ISO + ICI118551 group (55 nmol/L ICI118551 perfusion for 5 min before and throughout ISO treatment), ICI118551 group (55 nmol/L ICI118551 perfusion for 20 min). After these treatments, all hearts were followed by 30 min of no-flow ischemia and 30 min of reperfusion. A computer-based electrophysiological recorder system was used to measure changes of the maximal rate of pressure increase in systole phase (+dp/dt(max)), maximal rate of pressure decrease in diastole phase (-dp/dt(max)), and difference of left ventricular pressure (ΔLVP). Then cardiomyocytes from these hearts were isolated by 5 min of Ca(2+)-free buffer perfusion and 25 min of collagenase perfusion. The ventricles were chopped and filtered. The myocytes were resuspended in KB buffer. The contraction and the viability of cardiomyocytes were measured. Lactate dehydrogenase (LDH) concentration in coronary effluent was assayed with assay kit. The results showed that both IP and ISO significantly increased the values of ±dp/dt(max), ΔLVP, the contraction and viability of cardiomyocytes, shortened the time-to-peak contraction (TTP), and decreased the release of LDH in coronary effluent. ICI118551, a selective β(2)-AR antagonist, blocked these effects. Either the time-to-50% relaxation (R(50)) or the time-to-100% relaxation (R(100)) had no significant differences between groups. Our results indicate that the cardioprotection of IP was mediated by β(2)-AR in isolated rat hearts subjected to I/R injury.
Animals ; Heart ; physiology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Myocardial Contraction ; Myocardial Reperfusion Injury ; metabolism ; Myocytes, Cardiac ; physiology ; Perfusion ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-2 ; metabolism ; Reperfusion Injury

Chylothorax is an uncommon disease where fatty fluid accumulates within the chest cavity. Conservative management, including repeated thoracentesis or pleurodesis, seems to be suitable to most cases. Herein, we present a case of efficacious pleurodesis by intrapleural injection of Sapylin, a streptococcus preparation, for the treatment of chylothorax. A 52-year-old non-smoking female farmer was diagnosed as idiopathic chylothorax after we ruled out possible causes including chest trauma, lymphoma, lung cancer, filariasis, tuberculosis, and etc. Two-time intra-thoracic injection of 3 Klinische Einheit (KE) Sapylin achieved rapid and effective control of chylothorax with no severe side effects. Sapylin may facilitate pleurodesis by producing a strong inflammatory response.
Bacterial Vaccines ; therapeutic use ; Biological Products ; therapeutic use ; Chylothorax ; drug therapy ; pathology ; Female ; Humans ; Middle Aged ; Streptococcus ; chemistry ; Tomography, X-Ray Computed

OBJECTIVETo further study the role of human cytomegalovirus (HCMV) infection in the pathogenesis of the type 2 diabetes mellitus.

METHODSHCMV DNA levels in sera from 29 type 2 diabetic patients and 23 controls were measured by quantitative polymerase chain reaction (PCR), and comparative analyses was made between HCMV DNA and T cell subsets, blood glucose (BG), insulin (Ins) and C peptide (C-P).

RESULTSThe levels of HCMV DNA were (1.81+/-1.67) x 10(8) copies/ml for type 2 diabetic patients, a level significantly higher than that (5.50+/-4.30) x 10(7) copies/ml of controls. The percentage of CD8 for type 2 diabetic patients with positive HCMV DNA was 29.53%+/-2.00%, being much higher than that for controls (27.13%+/-4.12%), while the ratio of CD4/CD8 1.24+/-0.05 was significantly lower than that 1.41+/-0.10 of controls. Fasting C-P of type 2 diabetic patients with positive HCMV DNA was far lower than that of those with negative HCMV DNA, but the differences of BG and Ins between the two groups were not significant.

CONCLUSIONActive HCMV infection of type 2 diabetic patients may suppress cellular immunity and its influence on the pathogenesis of the type 2 diabetes mellitus should be further studied.

Adult ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; complications ; immunology ; DNA, Viral ; blood ; Diabetes Mellitus, Type 2 ; immunology ; virology ; Female ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; T-Lymphocyte Subsets

In order to explored the change of PML/PML-RARalpha protein during tetraarsenic tetrasulfide (As4S4) treatment, acute promyelocytic leukemia (APL) cells from a group of newly diagnosed APL patients were examined by indirect immunofluorescence staining with anit-PML monoclonal antibody. The results showed that all samples typically presented many microspeckle signals throughout the nucleus before treatment. The redistribution occurred as early as on the second day after As4S4 treatment, which revealed loss of microspeckles with the presentation of a few large speckles. Anti-PML staining also emerged in the perinuclear cytoplasm. At last, microspeckles and large speckles all disappeared. When the therapy was combining all-trans-retinoic acid (ATRA) with As4S4, similar results were obtained. However, APL cells from patients treated with ATRA alone performed totally different appearance, presenting microspeckles and large speckles at the same time, followed with entirely large speckles. The conclusion is that As4S4 makes redistribution of PML/PML-RARalpha protein in leukemic cells from APL patients during the treatment, which is quite different from that during the treatment of ATRA.
Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Arsenicals ; therapeutic use ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; In Situ Nick-End Labeling ; Leukemia, Promyelocytic, Acute ; drug therapy ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Oncogene Proteins, Fusion ; analysis ; Promyelocytic Leukemia Protein ; Transcription Factors ; analysis ; Tretinoin ; therapeutic use ; Tumor Suppressor Proteins

AIMTo investigated the effect of estrogen on global myocardial ischemia/reperfusion (I/R) injury in ovariectomized (Ovx) rats.

METHODSSprague-Dawley rats were randomly repartitioned into three groups including sham-operated(Sham), ovariectomized (Ovx), or ovariectomized and then given 17beta-estradiol (Ovx + E2). Hearts were excised, mounted on the Langendorff. After the initial stabilization period, all of the three group hearts were randomly divided into normal perfusion subgroup (Control) and I/R perfusion subgroups. Control, perfused for 60 min after stabilization. I/R perfusion subgroups divided into 10 min I + 30 min R, 20 min I + 30 min R, 30 min I + 0 min R, 30 min I + 5 min R, 30 min I + 15 min R and 30 min I + 30 min R. And then, every group hearts were isolated into the single cardiomyocyte. The cardiomyocytes basal contraction and isoproterenol(ISO) stimulation contraction were measured. The viability and yield of cardiomyocytes were counted. LDH and CK concentrations in coronary effluent were assayed with assay kit.

RESULTSThe viability and yield of cardiomyocytes were significantly decreased in the conditions of 30 min ischemia followed by different times of reperfusion. The releases of LDH and CK in coronary effluent were significantly increased in the conditions of 30 min ischemia followed by different times of reperfusion. Except the 10 min and 20 min ischemia, the releases of LDH and CK were significantly increased in Ovx during I/R. Ovx + E2 could abate the heart injury through decreasing the releases of LDH and CK. Besides the control and the 10 min I + 30 min R groups, the myocardial basal and ISO stimulation contraction were higher from Ovx than Sham, and the effect was reversed by Ovx + Ez.

CONCLUSIONThe results indicate estrogen plays a cardioprotective role in global myocardial ischemia/reperfusion injury in ovariectomized (Ovx) rats.

Animals ; Estradiol ; pharmacology ; Estrogens ; pharmacology ; Female ; Myocardial Contraction ; physiology ; Myocardial Ischemia ; physiopathology ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; metabolism ; physiology ; Ovariectomy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.
Acetonitriles ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Formates ; chemistry ; Tandem Mass Spectrometry ; methods ; Vitaceae ; chemistry ; Water ; chemistry

AIMTo study the antidiabetic effects of cortex Moutan polysaccharide-2b (CMP-2b) in type 2 diabetes mellitus (T2DM) rats.

METHODSThe T2DM model rats were induced by a single intravenous injection of low dose streptozotocin (STZ) and intake of high sucrose-fat diet. CMP-2b was given to T2DM rats daily through gavage for 4-5 weeks. The body weight, water and food intake, fasting blood glucose (FBG), glucose tolerance, plasma lipids, serum insulin, and insulin receptor (Ins R) were determined.

RESULTSOral administration of CMP-2b significantly decreased water and food intake, FBG, total cholesterol (Tch), and triglyceride (TG), improved the impaired glucose tolerance (IGT), and remarkably raised the number of low affinity InsR and insulin sensitivity index (ISI) in T2DM rats.

CONCLUSIONCMP-2b may be useful for treating T2DM and its complications.

Animals ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Diabetes Mellitus, Experimental ; drug therapy ; Drugs, Chinese Herbal ; chemistry ; Glucose Intolerance ; Hypoglycemic Agents ; isolation & purification ; therapeutic use ; Insulin ; blood ; Liver ; metabolism ; Male ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; therapeutic use ; Rats ; Rats, Wistar ; Receptor, Insulin ; metabolism

OBJECTIVETo investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.

METHODThe effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.

RESULTAfter 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.

CONCLUSIONCur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.

Cell Cycle ; drug effects ; radiation effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Curcumin ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; radiation effects ; Humans ; RNA, Long Noncoding ; genetics ; Radiation Tolerance ; drug effects