This study is to evaluate the effects of Shenmai injection on the temporal alterations of action potential (AP), early afterdepolarization (EAD) and delayed afterdepolarization (DAD) in papillary muscles. The action potentials were recorded by a glass electrode. APD at 90% repolarization (APD9 ) was measured, and spontaneous EAD and DAD were observed. The results show APD90 was significantly prolonged in model group compared with sham-operated group, whereas it was remained unchanged in Shenmai injec- tion treatment group and amiodarone group. The spontaneous EADs and DADs were frequently visible in model group. In conclusion, EAD, DAD and trigger activities increase gradually during pathological progression of rat cardiac hypertrophy, and Shenmai injection could improve the action potential change in rat cardiac hypertrophy.
Action Potentials ; drug effects ; Animals ; Blood Pressure ; drug effects ; Cardiomegaly ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Heart Ventricles ; drug effects ; physiopathology ; Injections ; Male ; Papillary Muscles ; drug effects ; physiopathology ; Rats ; Rats, Sprague-Dawley

Proteins encoded by regions of difference (RD) of Mycobacterium tuberculosis (MTB) constitute a potential source of specific antigens for vaccine development and immunodiagnosis .In the present study ,four genes named nrdF1 , pe_pgrs35 ,rv1985c ,and rv1986 from RD2 of MTB were cloned and overexpressed in E .coli with the induction of IPTG .Western blotting assay showed that these recombinant fusion proteins could well react with anti-His tag monoclonal antibody ,which in-dicated their good immunoreactivity .The serodiagnosis potential applications of these four proteins in bovine tuberculosis were further evaluated .An indirect ELISA assay was established by using fusion proteins as coating antigens for detection of their specific antibodies in bovine sera .The positive rates were 7 .35% in NrdF1 ,22 .06% in PE_PGRS35 ,16 .18% in Rv1985c and 16 .18% in Rv1986 respectively .All the results suggest that these fusion proteins have the potential application in serodiagnosis of bovine tuberculosis .

BACKGROUND:In recent years, increasing research emphasizes the puncture position of the foramen ovale in skul , but most of the positin methods require a higher personal experience of surgeons and lack of individualized quantitative parameters. OBJECTIVE:To establish a visualized digital model of the foramen ovale in skul , explore the reasonable puncture path and puncture depth of percutaneous treatment of foramen ovale puncture for trigeminal neuralgia, and develop the individualized treatment of trigeminal neuralgia. METHODS:Head CT images from healthy adult male volunteers were obtained and were input into three-dimensional reconstruction software MIMICS 10.01, the three-dimensional visualized models of the skul and skin were established. Using the models, the puncture path of the foramen ovale was designed and the preliminary model of the puncture locator was plotted. RESULTS AND CONCLUSION:The three-dimensional visualized digital model of the foramen ovale puncture path was established with CT scan images by using MIMICS software, which provides reliable anatomical data for clinical teaching and lays the groundwork for the simulation of puncture surgery. On the three-dimensional models, the“needle points”,“target points”, and“midpoint”were determined, and the triangle consisted of the three points was regarded as“positioning plane”. Using these parameters, the positioning instrument is characterized by simple structure, convenient operation, high positioning precision and short period of exposure to radiation, it simulates the foramen ovale puncture needle depth and needle direction in a precise and individualized manner.

In order to establish the stable andreliable ISSR-PCR System of Lysimachia christinae, L16 (4(5)) orthogonal design, which based on 7 levels of single factor experiment, were used in this study. The variance analysis was carried out by SPSS 19.0, and 5 main factors affecting the reaction system were optimized in 4 levels. The best annealing temperature was selected by the optimized reaction system. And the stability and reliability of this system was tested by 23 samples from different origins. The results showed that the five factors (DNA template, primer, dNTP, Mg2+ and Taq enzyme) were the most impacts on the amplified results of ISSR-PCR of L. christinae. The order of the influence was: primer > Taq enzyme > DNA template > Mg2+ > dNTP. The optimal system, which was determined by multiple comparison on different levels of each factor, was total volume of 25 microL, including DNA template 60 ng, primer 0.3 micromol x L(-1), dNTP 0.2 mmol x L(-1), Mg2+ 1.8 mmol x L(-1), Taq enzyme 1.25 U. The optimal system was stable and reliable tested by 23 samples from different origins. This study lays the foundation for genetic diversity analysis, fine varieties selection and molecular identification of L. christinae, and provides reference for optimization on ISSR-PCR system of other speciesin future.
DNA Primers ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Primulaceae ; classification ; genetics ; Quality Control

Adult ; Fatty Liver ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Pregnancy ; Pregnancy Complications ; diagnosis ; diagnostic imaging ; Ultrasonography ; Young Adult

The present study investigated the material basis of Urtica fissa for the inhibition of benign prostatic hyperplasia(BPH). The active fractions were screened, and the extracts of dichloromethane and ethyl acetate exhibited significantly inhibitory activities against 5α-reductase in vitro and BPH in model rats. The chemical constituents in the active fractions were systematically investigated, and 28 compounds were obtained, which were identified as lobechine methyl ester(1), dibutyl-O-phthalate(2), 1-monolinolein(3), epipinoresinol(4), 5-hydroxy-3,4-dimethyl-5-pentanyl-2(5H)-furanone(5), E-7,9-diene-11-methenyl palmitic acid(6), evofolin B(7), ficusal(8), threo-2,3-bis-(4-hydroxy-3-methoxyphenyl)-3-ethoxypropan-1-ol(9), α-viniferin(10),(9R,7E)-9-hydroxy-5,7-mengatigmadien-4-one-9-O-β-D-glucopyranoside(11), indole-3-carboxaldehyde(12), p-hydroxy ethyl cinnamate(13), benzyl alcohol-O-β-D-glucoside(14), L-methionine(15), 4-methoxyaniline(16), 6-aminopurine(17), 8'-acetyl oilvil(18), 4-methoxyl-8'-acetyl oilvil(19), vanillic acid(20), β-hydroxypropiovanillone(21), 7-hydroxy-6-methoxycoumarin(22), p-hydroxybenzaldehyde(23), pinoresinol(24), erythro-1,2-bis-(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(25), urticol(26), urticol-7-O-β-D-glucopyranoside(27), and lobechine(28). Compounds 1-17 were isolated from U. fissa for the first time. Meanwhile, compound 1 was a new natural product. Compounds 10, 11, 19, 21, and 27 exhibited significant inhibitory effects on 5α-reductase.
Animals ; Plant Extracts/pharmacology* ; Prostatic Hyperplasia/drug therapy* ; Rats ; Urticaceae/chemistry*

The aim of the present study is to investigate the change of thioredoxin (Trx) system in myocardial tissue of type 2 diabetic rats after myocardial injury and the underlying mechanism. Adult Sprague Dawley rats were randomly divided into two groups: normal control (NC) group and diabetes (DM) group. Rats in DM group were subjected to high-sugar, high-fat diet and streptozotocin (STZ) injection. Rats in NC group were only given normal diet and equal amount of citric acid buffer injection. At week 1, 2, 4, 12, 21 after STZ injection, plasma glucose concentration and the concentrations of insulin, creatine kinase MB (CK-MB), cardiac troponin I (cTnI) in serum were measured. Myocardial Trx and thioredoxin reductase (TR) activities, as well as caspase-3 activity, were determined by respective assay methods. Protein and mRNA levels of Trx, TR, Trx interacting protein (TXNIP) were determined by Western blot and real time PCR, respectively. The results showed that type 2 diabetic rat model was successfully established at week 1 after STZ injection, and myocardial injury was induced from week 2. Moreover, caspase-3 activity was significantly increased at week 4, 12 in diabetic rats. The activities of myocardial Trx and TR in diabetic rats was decreased from week 2, and continually aggravated as the disease developed. Compared with those in NC group, the mRNA levels of Trx1, Trx2, TR1, TR2 in DM group decreased at week 4, and then increased in week 12. In DM group, the protein levels of Trx1, Trx2, TR1 and TR2 increased significantly at week 12. The mRNA expressions of myocardial TXNIP in diabetic rats were significantly increased at week 4, 12, 24 and protein expression was increased at week 12. These results suggest diabetes can decrease myocardial Trx, TR activity, inducing myocardial cell apoptosis and heart injury. The inhibitory effect of diabetes is mainly associated with TXNIP up-regulation and Trx nitration.
Animals ; Apoptosis ; Carrier Proteins ; metabolism ; Caspase 3 ; metabolism ; Creatine Kinase, MB Form ; blood ; Diabetes Mellitus, Experimental ; physiopathology ; Diet, High-Fat ; Insulin ; blood ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Thioredoxin-Disulfide Reductase ; metabolism ; Thioredoxins ; metabolism ; Troponin I ; blood ; Up-Regulation

OBJECTIVETo evaluate the CD4+CD25+ regulatory T cells (Treg) and other lymphocyte subsets in the peripheral blood of patients with advanced lung adenocarcinoma.

METHODSPeripheral blood samples were obtained from 64 patients with advanced lung adenocarcinoma (case group) and analyzed by flow cytometry. The ratios of CD4+CD25+Treg T cells and other T lymphocyte subsets in peripheral blood were compared with those from 33 healthy controls (control group).

RESULTSThe percentages of CD3+ and CD3+CD4+ were (66.5±11.0)% and (37.7±10.6)% respectively in the peripheral blood of the case group, which were significantly lower than those [(72.0±6.0)% and (42.0±6.4)%] in the control group (t=-3.2,-2.4; P=0.020, 0.015, respectively). The ratio of CD4+ CD25+ Treg cells in case group (10.5±4.0)% was significantly higher than that [(8.4±3.5)%] in the control group (t=-2.2, P=0.013). CD4+/CD8+ value of case group (1.4±0.8) was significantly lower than that (1.8±0.7) in control group(t=-2.2, P=0.029). CD3+CD8+, CD8+CD28-, and CD8+CD28+ showed no significant differences (all P>0.05). Smoking, differentiation grade, and size of the tumor showed no association with the function damage of T lymphocyte subsets, while the carcino-embryonic antigen level did.

CONCLUSIONSIn patients with advanced lung adenocarcinoma, Treg increases and CD4+/CD8+ decreases, suggesting remarkably suppressed immune functions. However, more research is warranted to validate the association of T cells subset dysfunction with smoking, differentiation grade, and size of tumor.

Adenocarcinoma ; immunology ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Lung Neoplasms ; immunology ; Male ; Middle Aged ; Risk Factors ; T-Lymphocyte Subsets ; immunology

iASPP can prompt the cell proliferation and inhibit the apoptosis of many cells. There are putative binding sites of transcription factor GATA-2 upstream of iASPP transcription start site. GATA-2 plays an important role in the proliferation and differentiation of hematopoietic stem cells (HSC) and progenitors. This study was aimed to explore the role of GATA-2 protein in iASPP gene transcription. Firstly, the expression of iASPP and GATA-2 protein in some leukemia cell lines was detected by Western blot. Second, The expressive vector of pCMV5-GATA2 and the luciferase reporter vectors containing possible binding sites of GATA-2 were constructed and co-transfected into HEK293 and CV-1 cells. Then the luciferase activity was assayed by luminometer. Also, ChIP assays were performed to further confirm the specific binding of GATA-2 to iASPP promoter. The results showed that GATA-2 was overexpressed in most cell lines with high level of iASPP. GATA-2 exhibited a significant effect on luciferase activity of reporter gene iASPP and in a dose-dependant manner. The relative luciferase activity was up-regulated to about two-fold of the empty vector control when the transfection dose of pCMV5-GATA2 plasmid was increased to 100 ng. While the effect was more significant in CV-1 cells and showed a 6.7-fold increase. The ChIP assay demonstrated the in vivo specific binding of GATA-2 to iASPP. The binding sites of GATA2 were located between nt -361 ∼ -334 in upstream of iASPP gene transcription start site. It is concluded that transcription factor GATA-2 can bind with the cis-regulatory region of the iASPP promoter and up-regulate iASPP expression.
Animals ; Cell Line ; Cercopithecus aethiops ; GATA2 Transcription Factor ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; K562 Cells ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transcriptional Activation ; Transfection

Adenovirus transfection technique was used in the current study to show if thioredoxin-interacting protein (TXNIP) overexpression can induce cell apoptosis and injury in H9C2 cardiomyocytes cultured in normal glucose condition. And the mechanisms were then investigated. Briefly, H9C2 cardiomyocytes in logarithmic growth phase were randomly divided into three groups: normal cultured group, empty adenovirus vector group (Ad-eGFP) and TXNIP overexpression group (Ad-TXNIP-eGFP). All cells were cultured in DMEM containing normal concentration of glucose (5 mmol/L) and lipid. 72 h after adenovirus transfection, cells and culture mediums were collected for further assay. The results showed that Ad-eGFP and Ad-TXNIP-eGFP adenovirus transfected H9C2 cells successfully, and the transfection efficiency reached the peak at 72 h. Compared with Ad-eGFP group, Ad-TXNIP-eGFP transfection significantly increased TXNIP mRNA (P < 0.05) and protein expression level (P < 0.01). TXNIP overexpression induced remarkable cell apoptosis and injury as evidenced by increased caspase-3 activity (P < 0.05), apoptotic rate (P < 0.01) and LDH activity (P < 0.01). To further analysis the mechanisms of TXNIP-induced cell apoptosis, we also determined Trx activity, Trx related free radical injury and p38 kinase activation, which are involved in free radical induced apoptosis. The results showed that, compared with those in Ad-eGFP group, Trx activity was significantly decreased (P < 0.01), while malondialdehyde (MDA), 3-nitrotyrosine contents and p38 kinase activity were significantly increased (P < 0.01) in TXNIP overexpression group. These results suggest that TXNIP overexpression alone can induce severe apoptosis and injury in H9C2 cardiomyocytes even they are cultured in normal glucose and lipid concentration conditions. The mechanism involved is that overexpressed TXNIP can bind and inhibit Trx, impairs its antioxidative and antiapoptotic function, and then increases free radical induced injury and p38 kinase dependent apoptosis.
Adenoviridae ; genetics ; Animals ; Apoptosis ; Carrier Proteins ; genetics ; metabolism ; Caspase 3 ; metabolism ; Cell Line ; Genetic Vectors ; Myocytes, Cardiac ; cytology ; Rats ; Thioredoxins ; metabolism