Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

Abstract: This study is to improve the affinity of scFv-AK404R against VEGFR2. The secondary mutational library was constructed by hydrophilic shuffling in CDR3 region of the heavy chain. VEGFR2-specific screening was performed by phage display technology and the protein of mutants was expressed in periplasm of E.coli HB2151 and purified by affinity chromatography. The affinity constant of scFvs was measured by competitive ELISA, and the structure of scFvs was analyzed by bioinformatics. The result showed that a library with 6.4x10(5) scFv members was established by electro-transformation. Two mutated clones with high absorbance value were isolated after screening. After purification by affinity chromatography, electrophoretically pure scFv proteins were obtained. The competitive ELISA showed that the affinities of WZ01 and WZ02 were three times higher than that of the parental AK404R, and bioinformatics analysis showed that the enlarged contact surface and fitted closely with KDR3 surface may be the reasons for improved affinity. These results suggest that introducing hydrophilic amino acids to the heavy chain CDR3 region is an effective approach to improve the affinity of scFv.
Amino Acid Sequence ; Antibody Affinity ; Chromatography, Affinity ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Hydrophobic and Hydrophilic Interactions ; Peptide Library ; Single-Chain Antibodies ; genetics ; immunology ; isolation & purification ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; metabolism

To study clinical outcome of G-CSF-mobilized allo-peripheral blood stem cell (PBSC) and allo-bone marrow (BM) transplantation for patients with leukemia, donors were injected G-CSF 8-10 microg/(kg.d) for 5 days, PBSC were collected on day 4-5 and G-CSF mobilized BM was extracted on day 7. Conditioning regimen consisting of fludarabine, busulfan and cyclophosphamide. The results showed that transplanted cells in all patients were engrafted, the median days of neutrophil exceeding 0.5 x 10(9)/L and platelet exceeding 20 x 10(9)/L were 10.2 days (range 9 - 12 days) and 12.5 days (range 12 - 14 days), respectively. Patients were monitored up to 100 days, 4 of 12 patients (33.3%) developed II aGVHD, 10 of 12 patients (83.3%) developed limited cGVHD. The median follow-up duration was 5 months. Two patients died, the others were alive in disease-free situation. In conclusion, allogeneic transplantation of G-CSF mobilized PBSC plus BM was safe and effective treatment for patients with leukemia, the therapy provides rapid and sustained engraftment without increase in incidence of aGVHt and cGVHD.
Adolescent ; Adult ; Bone Marrow Transplantation ; methods ; China ; Female ; Follow-Up Studies ; Graft vs Host Disease ; diagnosis ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Leukemia ; therapy ; Male ; Peripheral Blood Stem Cell Transplantation ; methods ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo investigate the material foundation of the fusion of bcr and abl genes, and to explore the pathogenesis of chronic myeloid leukemia.

METHODSBy FISH combined with laser confocal scanning microscopy, the three-dimension (3D) distribution of bcr and abl genes in the interphase nuclei of normal and irradiated IM-9 cells was studied in each cell cycle phases.

RESULTSabl and bcr genes distributed non-randomly in the interphase nuclei of IM-9 cells. abl gene preferably located at the outer layer and bcr near the core of the nucleus. The two genes were drawn near each other most in G(0) phase. The relative distance between the homologous genes was greater at proliferation phase than at quiescence phase. After irradiation, the relative distances from the two genes to the core and between the two genes were shortened, with the shortest distance between the two genes in S phase.

CONCLUSIONIrradiation could change the 3D-distribution of abl and bcr genes in the interphase nuclei of IM-9 cell and accelerate them to draw near each other.

Cell Nucleus ; genetics ; radiation effects ; ultrastructure ; Cells, Cultured ; Female ; Fusion Proteins, bcr-abl ; genetics ; radiation effects ; Gene Fusion ; radiation effects ; Genes, abl ; genetics ; radiation effects ; Humans ; In Situ Hybridization, Fluorescence ; Interphase ; genetics ; radiation effects ; Lymphocytes ; ultrastructure ; Microscopy, Confocal ; Proto-Oncogene Proteins c-bcr ; genetics ; radiation effects

BACKGROUND: Primary pulmonary malignancies (PPMs) and non-pulmonary malignancies (PNPMs) may result in airway stenosis requiring stenting. This study aimed to compare and evaluate the clinical features and stent placement outcomes of airway stenosis caused by PPMs and PNPMs. METHODS: A total of 141 patients with malignant airway stenosis who underwent Micro-Tech stent placements between January 2004 and October 2017 at Department of Respiratory Medicine, Beijing Tian Tan Hospital, Capital Medical University were divided into PPM (n = 100) and PNPM groups (n = 41). Patients' clinical features and stent placement outcomes were collected and analyzed. Chi-square test was used to compare the categorical variables, while independent- or paired-sample t test was used to compare the continuous variables. RESULTS: There were no significant differences in age, sex, treatment history, respiratory symptoms, and incidence of obstructive pneumonia between groups. Multiple airway involvement (63.0% vs. 31.7%; χ = 11.459, P = 0.001) and atelectasis (17.0% vs. 2.4%; χ = 5.536, P = 0.019) were more common in the PPM group, while extraluminal obstruction (24.4% vs. 6.0%; χ = 8.033, P = 0.005) was more common in the PNPM group. Before stenting, the American Thoracic Society Dyspnea Index (ADI) and Karnofsky Performance Scale (KPS) scores showed no significant differences between groups (all P > 0.05). After stenting, a satisfactory rate of symptom improvement was achieved in both groups (98.0% and 100.0% in the PPM and PNPM groups, respectively; χ = 0.016, P = 0.898); ADI and KPS scores, which showed no significant differences between groups (all P > 0.05), were significantly improved in each group (all P < 0.001). Complications after stenting could be effectively managed using bronchoscopic procedures. CONCLUSIONS Among cases of malignant airway stenosis requiring stenting, those caused by PPM are more likely to involve multiple airways and are associated with atelectasis, while those caused by PNPM are more likely to cause extraluminal obstruction. Micro-Tech stent placement has the same immediate effect in terms of improvement in respiratory symptoms and performance status for both malignant airway stenosis caused by PPM and that caused by PNPM.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lung Neoplasms ; complications ; Male ; Middle Aged ; Stents ; adverse effects ; Tracheal Stenosis ; etiology ; therapy

OBJECTIVETo investigate the effect of irbesartan on the renal expressions of advanced glycation end products (AGEs) and their receptor (RAGEs) in rats with early diabetic nephropathy (DN) and the renoprotection mechanism of irbesartan.

METHODSRat DN models established by a single injection of streptozotocin were randomly divided into the model group and irbesartan treatment group. With normal rats as the control, all the rats received daily gavage for 8 weeks. The 24-h urinary protein excretion and contents of AGEs in the serum and kidney tissues were measured. The expressions of RAGEs and RAGEs protein and mRNA in the kidney tissues were detected by immunohistochemistry and reverse transcription-polymerase chain reaction, respectively. The pathological changes of the kidney were also assessed microscopically.

RESULTSIrbesartan significantly reduced the 24-h urinary protein excretion and the contents of AGEs in the serum and kidney tissues of DN rats, resulting also in decreased expressions of RAGEs and RAGEs protein and mRNA levels in the kidney. The treatment obviously alleviated the pathological changes in the kidney of the DN rats.

CONCLUSIONIrbesartan offers renoprotection against DN possibly by reducing the serum and renal contents of AGEs and inhibiting the renal mRNA expressions of RAGEs and RAGEs.

Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Animals ; Biphenyl Compounds ; therapeutic use ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; drug therapy ; metabolism ; Glycation End Products, Advanced ; genetics ; metabolism ; Kidney ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; genetics ; metabolism ; Tetrazoles ; therapeutic use

In order to explore the activity of a peptide containing rat sodium pump α2 subunit M1-M2 extramembrane fragment (RES2 derivative) in vitro, the peptide (Leu-Ala-Ala-Met-Glu-Asp-Glu-Pro-Ser-Asn-Asp-Asn-Gly-Gly-Gly-Ser) was synthesized by peptide synthesizer with Fmoc method and purified by high performance liquid chromatography (HPLC). Its binding activity was identified by radioligand-receptor binding assay (RRA) and its bioactivity was measured by erythrocyte (86)Rb uptake. The results of saturation binding experiment and competitive binding experiment showed that the synthesized RES2 derivative had the capability to bind to (3)H-ouabain. The dissociation constant (K(d)) was 38.46 nmol/L and IC(50) was 6.353 nmol/L. Erythrocyte (86)Rb uptake experiment showed that the RES2 derivative blocked the inhibitory effect of ouabain on the sodium pump on erythrocyte membrane in a dose-dependent manner. The results showed that the RES2 derivative is capable of binding to ouabain and improving the activity of sodium pump on erythrocyte membrane, suggesting that the RES2 derivative might become an effective antihypertensive drug in the future.
Amino Acid Sequence ; Animals ; Chromatography, High Pressure Liquid ; Erythrocyte Membrane ; drug effects ; Ouabain ; pharmacology ; Peptide Fragments ; metabolism ; Rats ; Sodium-Potassium-Exchanging ATPase ; metabolism

The near infrared (NIR) spectroscopy technique has been applied in many fields because of its advantages of simple preparation, fast response, and non-destructiveness. We investigated the potential of NIR spectroscopy in diffuse reflectance mode for determining the soluble solid content (SSC) and acidity (pH) of intact loquats. Two cultivars of loquats (Dahongpao and Jiajiaozhong) harvested from two orchards (Tangxi and Chun'an, Zhejiang, China) were used for the measurement of NIR spectra between 800 and 2500 nm. A total of 400 loquats (100 samples of each cultivar from each orchard) were used in this study. Relationships between NIR spectra and SSC and acidity of loquats were evaluated using partial least square (PLS) method. Spectra preprocessing options included the first and second derivatives, multiple scatter correction (MSC), and the standard normal variate (SNV). Three separate spectral windows identified as full NIR (800approximately2500 nm), short NIR (800approximately1100 nm), and long NIR (1100approximately2500 nm) were studied in factorial combination with the preprocessing options. The models gave relatively good predictions of the SSC of loquats, with root mean square error of prediction (RMSEP) values of 1.21, 1.00, 0.965, and 1.16 degrees Brix for Tangxi-Dahongpao, Tangxi-Jiajiaozhong, Chun'an-Dahongpao, and Chun'an-Jiajiaozhong, respectively. The acidity prediction was not satisfactory, with the RMSEP of 0.382, 0.194, 0.388, and 0.361 for the above four loquats, respectively. The results indicate that NIR diffuse reflectance spectroscopy can be used to predict the SSC and acidity of loquat fruit.
Eriobotrya ; chemistry ; Hydrogen-Ion Concentration ; Least-Squares Analysis ; Spectroscopy, Fourier Transform Infrared ; methods ; Spectroscopy, Near-Infrared ; methods

OBJECTIVETo investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligand on angiotensin II (AngII)-induced endothelin-1 (ET-1) and NO secretion by endothelial cells in comparison with AngII type I receptor (AT1R) antagonist losartan, so as to reveal the relationship between PPAR gamma and essential hypertension.

METHODSCultured human umbilical vein endothelial cells (HUVECs) were treated with AngII, PPAR gamma ligand troglitazone, AngII plus troglitazone, and AngII plus AT1R antagonist losartan, respectively, and the concentrations of NO and ET-1 in the cell culture supernatant were measured to evaluate the effects of troglitazone and losartan on AngII-induced NO and ET-1 production by human endothelial cells.

RESULTSTreatment of the HUVECs with troglitazone at 10 micromol/L and 50 micromol/L did not produce significant changes in ET-1 concentration in the cell culture supernatants, but significantly increased NO concentration as compared with the control group (P<0.05). Triglitazone at the concentration of 50 micromol/L significantly inhibited AngII (1x10(-6) mol/L)-induced ET-1 production (P<0.05), and at both 10 and 50 micromol/L, troglitazone inhibited the NO release-lowering effect of AngII in the endothelial cells (P<0.05). Both troglitazone and losartan inhibited AngII-induced ET-1 production by the endothelial cells, but losartan showed more potent effect (P<0.05). Similarly, both troglitazone and losartan inhibited decreased NO production in response to AngII treatment, and again losartan showed stronger effect (P<0.05).

CONCLUSIONPPAR gamma ligand troglitazone can inhibit AngII-induced ET-1 production enhancement and decreased NO release by the endothelial cells, but its effect is not so strong as losartan, suggesting that troglitazone modulates blood pressure not solely through AT1R pathway.

Angiotensin II ; metabolism ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Cell Line ; Chromans ; pharmacology ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; metabolism ; secretion ; Endothelin-1 ; secretion ; Gene Expression Regulation ; drug effects ; Humans ; Hypertension ; metabolism ; Immunohistochemistry ; Losartan ; pharmacology ; Nitric Oxide ; secretion ; PPAR gamma ; metabolism ; Receptor, Angiotensin, Type 1 ; metabolism ; Thiazolidinediones ; pharmacology