Total RNA was isolated from Siraitia grosvenorii fruit by the method of modified Trizol, according to S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD260/280, OD260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 mirog.g-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from S. grosvenorii. The GenBank accession number for this gene is KC907731. The SgDHAR gene contains a full-length cDNA of 1,252 bp including ORF of 819 bp and encodes a predicted protein of 272 amino acids. The molecular mass is 30.217 7 kD and the isoelectric point is 8.76. Homology comparison showed that it shared 87% nucleotide sequence homology with Cucumis sativus. Expression patterns using qRT-PCR analysis showed that SgDHAR was mainly expressed in fruit and stem, followed by flower, and was lowest in root, while the expression level was 6.83 times in triploid. T than that in diploid. Therefore, SgDHAR gene may be involved in abortion of triploid seedless S. grosvenorii.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Cucurbitaceae ; chemistry ; genetics ; DNA, Complementary ; genetics ; Flowers ; chemistry ; genetics ; Fruit ; chemistry ; genetics ; Molecular Conformation ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Roots ; chemistry ; genetics ; Plant Stems ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Protein Structure, Secondary ; RNA, Plant

This study was aimed to investigate the sensitizing effect of recombinant human PDCD5 (rhPDCD5) protein on chemotherapy of U937 cell line and its mechanism. The flow cytometry was performed to assess the changes of cell apoptosis and cell cycle influenced by rhPDCD5. Hochst 33258 staining was used to observe morphology of the apoptotic cells. The activity change of caspase-3 was detected to analyse the possible mechanisms of rhPDCD5-induced apoptosis. RT-PCR was performed to observe the expression level of drug-resistant genes. The results showed that the percentage of apoptotic cells and the activity of caspase-3 remarkably increased in U937 cells treated with rhPDCD5 combined with chemotherapeutic drug; the cell cycle arrest induced by anti-tumor drug was also enhanced when combined with rhPDCD5; meanwhile, the expression levels of drug-resistant genes were down-regulated in jointly treated U937 cells. It is concluded that the chemosensitizing mechanisms of rhPDCD5 are complex. rhPDCD5 may increase the cytotoxicity of anti-tumor drugs by promoting the caspase-3-related apoptosis, influencing cell cycle, decreasing the expression of drug-resistant genes and reversing drug-resistance.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Neoplasm Proteins ; pharmacology ; Recombinant Proteins ; pharmacology ; U937 Cells

Objective To observe the management and outcome of the cardiac tamponade patients during the ablation procedure using two different anticoagulation strategies. Methods All the patients developed tamponade during the ablation procedure were enrolled from January 2007 to December 2013 in our center. In group 1, warfarin was discontinued 3 to 5 days before the procedure and low molecular weight heparin (LMWH) was administered subcutaneously until ablation procedure day. In group 2, warfarin was not discontinued and the international normalized ratio INR was to maintained between 2 and 3. Results There were 27 patients (0.6%) developed cardiac tamponade out of a total 4487 patients received ablation in our center. The baseline clinical characteristics including age, left atrium, the heparin dose and ACT during the procedure had no signiifcant difference between the groups, except that the INR was higher in the group 2 (0.9±0.1 vs. 2.3±0.5, P<0.001). There was no signiifcant difference in the amount of pericardiac drainage between the two groups (365±222 ml vs. 506±300 ml, P=0.137). Two patients in group 1 patient (11.1%) and 1 in group 2 (11.1%) needed emergency surgical repair (P>0.999). The median hospital day was similar in the 2 groups [(9.6±3.3) d vs. (12.1±4.5) d, P=0.167]. There were no other serious complications and no hospital death. Conclusions Non-discontinuation of warfarin during peri-procedural catheter ablation of AF is not signiifcantly different to bridging with LMWH in the management and outcome of acute cardiac tamponade.

Objective To explore the effect of iodine excess on bone metabolism in experimental autoimmune thyroiditis (EAT) rats. Methods We selected 36 female Lewis rats with body weight of (131 ± 15)g,and divided them into 3 groups randomly: control group, EAT group and EAT + high iodine group, assuring 12 rats in every group. These rats were fed fodder with different concentration of iodine(0.9,0.9, 18.0 mg/kg), and rats in EAT group and EAT + high iodine group were immunized with pig thyroglobulin(pTG) and complete Freund's adjuvant(CFA) to create EAT model. After two weeks, the pathological changes of the thyroid tissues were observed,and the serum thyroid autoantibody[thyroglobulin antibody(TGAb) and thyroid microsomal antibody(TMAb)], the thyroid hormone levels[triiodo thyronine(T3) and thyrine(T4)] and some relevant data of bone metabolism[bone gla protein (BGP), tartrate-resistant acid phosphatase (TRAP), C-terminal propeptide of type Ⅰ procollagen (PICP),C-terminal telopeptide of type Ⅰ collagen (ICTP), insulin-like growth factor- 1 ( IGF- 1 ), osteoprotegerin (OPG) and receptor activator of NF-κB ligand (RANKL)] were measured. Results Inflammatory cell infiltration and local follicular structural damage were observed in the thyroid tissues of EAT rats in EAT group and EAT + high iodine group, and the pathological changes of EAT + high iodine group were mainly thyroid follicular expansion and integration. The level of serum TGAb, TMAb, T3 and T4 of EAT rats in EAT group and EAT + high iodine group[ (63.01 ± 12.36)%, (60.62 ± 11.24)%, (3.78 ± 1.43), (125.12 ± 16.00)pmol/L and (75.00 ± 15.44)%,(72.15 ± 15.00)%, (3.69 ± 0.91 ), (149.40 ± 20.67)pmol/L] were higher than those of the control group[ (4.47 ±1.04)%, (5.73 ± 1.01 )%, (0.75 ± 0.12), (76.91 ± 9.30)pmol/L, all P ＜ 0.05], and the level of serum TGAb,TMAb and T4 of EAT rats in EAT + high iodine group were higher than those of the EAT group(all P ＜ 0.05).The level of serum BGP, PICP and IGF- 1 in EAT group[ ( 1.70 ± 0.31 ), ( 11.31 ± 1.52) μg/L, (0.31 ± 0.06 ) mg/L]were lower than those of the control group[ (8.60 ± 0.33), (14.28 ± 3.10)μg/L, (1.16 ± 0.02)mg/L, all P ＜0.05], and the level of serum TRAP, ICTP, OPG and RANKL[ ( 19.88 ± 3.60)ng/L, (2.43 ± 0.82), (22.36 ± 2.80),( 1.35 ± 0.23 )μg/L] were higher than those of the control group[ ( 14.57 ± 3.56)ng/L, (0.50 ± 0.20), (1.61 ± 0.34),(0.10 ± 0.02)μg/L, all P ＜ 0.05]; compared with EAT group, the level of PCIP and OPG in EAT + high iodine group [ (8.03 ± 1.84), ( 16.80 ± 3.79)μg/L] were obviously decreased(all P ＜ 0.05). Conclusions The reinforcement of differentiation and maturation of osteoblast in the EAT rats results in the increasing of bone resorption. The activity of osteoblast and osteoclast of the EAT rats are inhibited by excessive iodine, showing a low conversion-type osteoporosis.

An automatic inspection system for biochip's print quality is presented in this paper. It consists of an automatic mechanical control, a CCD sensor for getting the image of PET boart, and the special computer software for image processing and recognition. Experimental results indicate that this system is capable of providing a precise and effective realtime inspection for biochips' print quality.
Biosensing Techniques ; instrumentation ; methods ; Equipment Design ; Image Processing, Computer-Assisted ; methods ; Microchip Analytical Procedures ; methods ; Pattern Recognition, Automated ; methods ; Quality Control ; Software

OBJECTIVETo study the protective effect of Ligustrazine on IR after local cerebral ischemia of rat.

METHODModels of rat IR after local cerebral ischemia were prepared by electrocagulation of the middle cerebral artery, and changes of serum insulin, tomer necrosis factor-alpha (TNF-alpha), plasma endothelin-1 (ET-1), nitric oxide (NO) and nitric oxide synthase (NOS) were observed 2 weeks after the ischemia.

RESULTLigustrazine could significantly reduce serum insulin (P < 0.01), the content of plasma ET-1 (P < 0.01) and serum TNF-alpha (P < 0.01), the activity of brain tissue NO and NOS (P < 0.01). The drug also increased insulin sensitivity indexes (ISI).

CONCLUSIONThe protective effects of Ligustrazion on IR cerebral ischemia may be related to decreasing ET-1 content in plasma, TNF-alpha content in serum, NO content and NOS activities in tissue.

Animals ; Brain Ischemia ; blood ; etiology ; Endothelin-1 ; blood ; Female ; Infarction, Middle Cerebral Artery ; complications ; Insulin ; blood ; Insulin Resistance ; Male ; Neuroprotective Agents ; pharmacology ; Pyrazines ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; Tumor Necrosis Factor-alpha ; metabolism

Objectives To investigate changes in serum levels of adiponectin,leptin and erythrocyte membrane insulin receptor among patients with gestational diabetes (GDM),and to study their relation to insulin resistance.Methods Fasting plasma glucose (FPG),fasting serum insulin (FINS), serum levels of adiponeetin and leptin,indices of lipid metabolism,2 h plasma glucose during oral glucose tolerance test (2 h PG),2 h serum insulin during oral glucose tolerance test (2 h INS),as well as number of erythrocyte membrane insulin receptors with high and low appetency and its constants,were determined in 40 patients with GDM and 34 controls with normal glucose tolerance.Insulin resistance index (IRI) was calculated.Results ① Serum levels of leptin and adiponectin were (11.7?2.8) ?g/L and (7.8?1.6) ?g/L,respectively,and number of high appeteney erythrocyte membrane insulin receptor (R_1) and low appetency erythrocytemembrane insulin receptor (R_2) was (43?9) / red cell and (2297?525) / red cell,respectively.Serum level of leptin was significantly higher in those with GDM than those of normal controls (P

OBJECTIVETo evaluate the safety and feasibility of video-assisted thoracic surgery (VATS) anatomic segmentectomy for pulmonary diseases.

METHODSBetween November 2009 and July 2011, 20 patients received consecutive VATS anatomic segmentectomy by single surgical group. There were 3 male and 17 female, aging from 32 to 81 years with a mean of 53 years. The patients included 12 cases of non-small cell lung cancer (NSCLC) (5 cases of bronchioloalveolar carcinoma, 6 cases of adenocarcinoma, and 1 case of squamous cell carcinoma), 7 cases of benign diseases (3 cases of bronchiectasis, 2 cases of inflammatory pseudotumor, 1 case of tuberculosis, and 1 case of sclerosing hemangioma) and 1 case of metastasis tumor. The locations of resected segments included 1 anterior segment, 3 posterior segments, 4 apical segments, and 2 superior segments in the right side; and 5 lingular segments, 3 trisegments, and 2 superior segments in the left side. Simultaneously, 3 patients with bronchiectasis underwent segmentectomy and lobectomy, 1 of 6 patients with adenocarcinoma underwent lingulectomy and thymectomy. The pathological TNM stages of 12 NSCLC patients were 9 cases of T1aN0M0, 1 case of T1bN0M0 and 2 cases of T2aN0M0.

RESULTSOf these 20 patients, the median operative time was 155 minutes (range, 120 to 235 minutes), the median blood loss was 50 ml (range, 10 to 600 ml), the median drainage duration was 3 d (range, 1 to 6 d), and the median hospital stay was 6 d (range, 3 to 9 d). One patient who had undergone lingulectomy had a 600 ml intraoperative bleeding from lingular artery, and the bleeding was controlled by suturing the rupture under VATS. Bloody sputum occurred in 2 patients, prolonged air leak occurred in one patient for 5 days, and one patient developed subcutaneous emphysema that spontaneously resolved. No mortality was observed for 30 days after the surgery.

CONCLUSIONSVATS anatomic segmentectomy is a feasible and safe technique with acceptable operative time, less blood loss, fewer complications, and shorter hospital stay.

Adult ; Aged ; Female ; Humans ; Lung Diseases ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods ; Thoracic Surgery, Video-Assisted ; Treatment Outcome

The aim of this study is to develop the recombinant adenovirus vaccine (rAdV) candidates containing neuraminidase (NA) gene of H5N1 influenza virus and test in BALB/c mice the effect of cell-mediated immunity. In this study, two kind of NA gene (WtNA gene, the wild type; Mod. NA gene, the codon-modified type) derived from H5N1 influenza virus (A/Anhui/1/2005) were cloned and inserted respectively into plasmid of adenovirus vector, then the rAdV vaccines candidates (rAdV-WtNA and rAdV-Mod. NA) were developed and purified, followed by immunization intramuscularly (10(9) TCID50 per dose, double injection at 0 and 4th week) in BALB/c mice, the effect of cell-mediated immunity were analysed at 5th week. Results indicated that: (i) NA protein expression was detected in two rAdV vaccines candidates by Western blotting; (ii) the rAdV-Mod. NA vaccine could elicit more robust NA specific cell-mediated immunity in mice than that of rAdV-WtNA vaccine (P = 0. 016) by IFN-gamma ELIspot assay. These findings suggested rAdV-Mod. NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.
Animals ; Blotting, Western ; Female ; Humans ; Immunity, Cellular ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; immunology ; Orthomyxoviridae Infections ; immunology ; virology ; Polymerase Chain Reaction ; Random Allocation ; Viral Proteins ; genetics ; immunology

BACKGROUNDNon-cement femoral stems are recognized in clinical use, but there are still some problems. The aim of this research was to make non-cement femoral stems to be press-fit with the medullary cavity.

METHODSTwenty-four healthy adult mongrel dogs were randomly divided into experimental and control groups. In the right hip joint, an artificial femoral bone replacement surgery was conducted. For the experimental group, the replacement surgery of hydroxyapatite (HA)-coated femoral stems was done, while autogeneous morselized bone was implanted into the medullary cavity. For the control group, morselized bone was not implanted. At postoperative 1, 3, 6 months, a test for interfacial shear characteristics was conducted in the MTS810 Tester. The comparison between the two groups' bone-prostheses in shear strength for their interface from shearing destruction was made. A histological observation to check prosthesis-bone interface contact ratios and bone growth was carried out.

RESULTSFor the experimental group, shear strength was 0.317 MPa in 1 month, 1.447 MPa in 3 months, and 1.621 MPa in 6 months. For the control group, shear strength was 0.195 MPa in 1 month, 1.023 MPa in 3 months, and 1.483 MPa in 6 months. The difference was statistically significant. Stereomicroscope-based observation showed that the number of trabecular bones in the experimental group was larger than that of the control group, and bone growth of the former group was better than that of the latter group. Inverted microscopic observation showed that the binding degree between the prosthesis and trabecular bone of the experimental group was higher than that of the control group. Comparatively, the experimental group's trabecular bone had more stromal cells.

CONCLUSIONSThe morselized bones can effectively improve the biological bonding strength and bone-contact ratios in the short term for the HA-coated femoral stem and accelerate the bonding process. The use of morselized autogenous bones played a good role in bone in-growth of the femoral bone stem surface.

Animals ; Biomechanical Phenomena ; Coated Materials, Biocompatible ; chemistry ; Dogs ; Durapatite ; chemistry ; Female ; Femur ; pathology ; surgery ; Male ; Osseointegration ; Random Allocation ; Shear Strength