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

· AIM:To investigate the correlation between retinal thickness (CSRT) in the macular region and glycosylated hemoglobin (HbA1c) of patients with type 2 diabetes mellitus.· METHODS:Totally 39 cases of patients with diabetes (77 eyes) who screened from May 2016 to March 2017 were selected,and were divided into two groups according to the levels of HbA1c,which the 24 cases (47 eyes) in the low HbA1c group (HbA1c＜8％) and 15 cases (30 eyes) in high HbA1c group (HbA1c≥8％).Other 22 cases of normal people (normal control group) and who for healthy physical examination were selected in the same period.Then,the correlation between HbA1c level and CSRT were analyzed by the Spearman correlation analysis.· RESULTS:In the high HbA1c group,HbA1c was (10.45±1.30)％,FBG was 10.67±1.64mmol/L and 2hPG was 15.98± 1.38mmol/L,which was higher than that in the low HbA1c group,and there was significant difference between the two groups (P＜ 0.05).The CSRT in the normal group was lower than the low HbA1c group and the high HbA1c group,and there was significant difference between the groups (P＜ 0.05).According to the analysis of the Spearman method,there was positive correlation between HbA1c and CSRT,macular volume,average macular thickness,FBG and those parameters,2hPG and them (P＜0.01).· CONCLUSION:The HbA1c level is associated with retinal thickening in the macular region of patients with diabetes,which could predict the severity of diabetic retinopathy and provide important guidance for prevention and treatment.

Food-borne pathogenic bacteria seriously threaten public health. Based on the mechanism of fluorescence resonance energy transfer (FRET), a ratiometric fluorescence biosensor was constructed by integration of Exo III-based signal amplification strategy. The Cy3 labeled R1-DNA firstly hybridized with Cy5 labeled R2-DNA to form duplex of R1/R2. Cy3 showed a low fluorescence response while Cy5 showed a high fluorescence response. The addition of target pathogenic bacterial gene (Lac Z gene) could de-hybridize the R1/R2,resulting in the fluorescence decreasing of Cy5 and the fluorescence recovering of Cy3. Under the assistance of Exo III, the signal change was further amplified. The detection of limit reached as low as 5.29 pmol/L. The linear detection range was from 10 pmol/L to 2000 pmol/L. The developed ratiomtric detection strategy significantly reduced the possibility of false-positive and false-negative detection results.

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

OBJECTIVETo investigate the effects of Shenkangwan on the expressions of angiotensin II (AngII) and its type I receptor (AT(1)R) and the renalprotection mechanism of Shenkangwan in rats with early diabetic nephropathy (DN).

METHODSThe rat models of DN established by a single injection of streptozotocin were randomly divided into 4 groups, namely the model group, Shenkangwan treatment group, irbesartan treatment group, and Shenkangwan and irbesartan treatment group, with normal rats as the control. All the rats received daily gavage for 8 weeks. The urinary protein quality in 24 h and plasma and renal contents of AngII were measured. The expressions of AT1R at the protein and mRNA levels in the kidney tissues were measured by immunohistochemistry and reverse transcription-polymerase chain reaction, respectively. The pathological changes of the kidney were observed microscopically.

RESULTSIn DN rats, Shenkangwan reduced the urinary protein quantity in 24 h and the contents of AngII in the plasma and kidney tissues, decreased the renal expressions of AT(1)R protein and mRNA, and alleviated the morphological damage of the kidney.

CONCLUSIONSShenkangwan offers renalprotection against DN probably by reducing the contents of AngII in the plasma and kidney tissues and inhibiting renal AT(1)R expressions.

Angiotensin II ; genetics ; metabolism ; Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Animals ; Diabetic Nephropathies ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Kidney ; metabolism ; Male ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism

Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.
Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; Cisplatin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Esophageal Neoplasms ; metabolism ; pathology ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Intramolecular Oxidoreductases ; metabolism ; Isotope Labeling ; Macrophage Migration-Inhibitory Factors ; metabolism ; Proteome ; metabolism ; Proteomics ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; Thioredoxins ; metabolism