OBJECTIVETo study the application value of normal sperm morphology on the outcomes of classic in vitro fertilization and embryo transfer (IVF-ET).

METHODSThis study included 659 infertile couples admitted to our center for IVF-ET. Based on the percentage of morphologically normal sperm (MNS), we divided the patients into groups A (n = 112, MNS < 2%), B (n = 180, MNS > or = 2 - < 4%), C (n = 74, MNS > or = 4 - < 5%), and D (n = 293, MNS > or = 5%), and compared the rates of fertilization, normal fertilization, embryos obtained, biochemical pregnancy, clinical pregnancy, implantation, and live birth among different groups.

RESULTSThe mean fertilization rate was significantly higher in groups C (71.90%) and D (72.89%) than in A (57.97%) and B (63.29%) (P < 0.05), with no remarkable differences either between A and B (P > 0.05) or between C and D (P > 0.05). The normal fertilization rate was also significantly higher in group D (57.16%) than in A (46.52%) and B (50.89%) (both P < 0.05) as well as in C (54.67%) than in A (P < 0.05). The rate of embryos obtained, too, was markedly higher in group D (55.62%) than in B (45.75%) (P < 0.05), but none with remarkable difference from other groups (all P > 0.05). There were no statistically significant differences among the four groups in the rates of biochemical pregnancy, clinical pregnancy, implantation, abortion, and live birth (all P > 0.05).

CONCLUSIONThe rate of MNS had some influence on IVF-ET, and 5% MNS exhibited a higher value than 4% MNS in predicting the outcomes of IVF.

Adult ; Embryo Implantation ; Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Spermatozoa ; cytology

Objective To analyze the distribution of pathogens in the genital tract of infertile female,and comparing traditional methods with simultaneous amplification and testing (SAT) in the detection of UU,CT,NG and MG.Methods 467 female infertility patients were selected from the reproductive center of Suzhou Hospital Affiliated to Nanjing Medical University between June and September 2016 to analyze the distribution of UU,CT,MG and NG.The age was between 20 to 48 years old (mean 31.52±6.83 years old).352 cases of female patients with assisted reproductive technology were selected,aged from 21 to 46 years old (mean 30.67±6.67 years old).The swabs were tested by traditional methods or SAT.The sensitivity and specificity of the methods in detecting the pathogens were evaluated according to the experimental results.Results Among the 467 infertile women,the number of UU positive cases was the highest,the positive rate was 62.53％ (292/467),the positive rate of CT was 1.93％ (9/467) and the positive rate of NG was 0.21％ (1/467),and the positive rate of MG was 1.71％ (8/467).UU infection rate was higher in infertile women than normal control group 23.81％ (25/105) (x2 =52.01,P＜0.01).352 cases of female patients with assisted reproductive technology were selected for further analysis.For UU detection,the positive rate of swab samples detected by liquid culture was 48.9％,while the positive rate detected by SAT was 63.9％.Obviously the positive rate of SAT was higher than that of liquid culture.Swab culture and SAT results were analyzed by paired x2 test (x2 =41.93,P＜0.01).The positive rate of CT SAT was 1.71％,and the positive rate of CT-latex method was 0.28 ％.There was significant difference between CT latex method and SAT (Fisher exact probabilistic method statistical analysis,P＜0.005),which indicated that SAT method had a higher sensitivity.The positive rate (1.7 ％) and sensitivity (100％) of SAT were also higher than that of traditional method.Conclusion UU was the most common pathogen in female reproductive tract pathogens,followed by CT and MG.The SAT method has higher sensitivity than the conventional method in detecting of UU and CT.

Aim To investigate the potential molecular basis of Epimedium and to clarify the molecular mechanism of Epimedium in the treatment of osteoporosis.Methods The method of network pharmacology was used to determine and screen the known compounds corresponding to Epimedium,and predict the drug-related gene/protein targets,and combined with bioinformatics,the specific target of osteoporosis prevention and treatment of Epimedium was determined,and then the signal pathway enrichment was analysed,the molecular mechanism of Epimedium in treatment of osteoporosis was further explored.Results A total of 130 corresponding compounds were retrieved in the TCMID database from Epimedium.According to the values of OB and DL,23 blood transfusion components were screened,in which 101 targets were obtained using the related target prediction technique.Through the secondary mining of the gene chip of GEO chip database,we obtained a total of 124 significantly different genes.And a total of 355 known target genes closely related to the development of osteoporosis were retrieved in the disease-related database.Using the cytoscape to construct and synthesize the protein-protein interaction network of active ingredients and diseases,221 key genes were screened out by network topological analysis.Using ClueGO analysis,it was shown that Epimedium was directly engaged in the signal pathways involved in the key nodes of osteoporosis,which were mainly related to the direct regulation of bone metabolism,such as Wnt signaling pathway,TGF-β signaling pathway and Notch signaling pathway,and that Epimedium regulated other signal pathways,such as PI3K-Akt signaling pathway,VEGF signaling pathway and thyroid hormone signaling pathway.Conclusions Epimedium has the characteristics of multiple targets and multicomponent in the treatment of osteoporosis,which could not only be directly involved in bone cell differentiation and regulation of the balance of osteogenesis and osteoclasts,but also affect and interfere with the bone microenvironment through other systemic systems,such as circulatory system,nervous system,which is consistent with the current mechanism of treatment of osteoporosis.

TSO18 gene was subcloned into the Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-TSO18 was transformed into P. pastoris GS115 by electroporation so that the plasmid will be integrated with chromosome of P. pastoris. The P. pastoris strains containing multi-copy recombinant were screened by G418 and induced by methanol. The expression product was analyzed by SDS-PAGE, Western blot, deglycosylation, and purified by Sephadex column, and was used to immunize mice. The results indicated that the target protein was efficiently expressed in P. pastoris, and glycosylated moderately, and had immunological activity. In a 5 liter fermentor, the expression level of the target protein was up to 2.54 mg/mL. These results will benefit for the development of genetically engineering vaccine.
Animals ; Antigens, Helminth ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Electroporation ; Gene Expression ; Genetic Vectors ; genetics ; Mice ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Taenia solium ; genetics ; immunology

Objective To study the impacts of the Three Gorges dam and change of water level on the survival of the local rodents, and to provide scientific basis to control the outbreak of rodent-borne diseases.Methods Four villages located around the Three Gorges dam were selected in the study. The mouse populations by using Elton night trapping method was monitored. Metallic spring traps were set for two consecutive nights. The mouse density and identified the mouse species was calculated. The mouse species indoor and outdoor, as well as the mouse density indoor and outdoor were compared. The impacts of water level in the dam and cleaning work on local mouse density were also analyzed. Results A total of 678 mice were caught in this study, 517 were caught indoor and 161 outdoor. Indoor dominant species was flavipectus; accounting for 36.49%(189/517), while outdoor was apodemus, reaching 56.88% (91/161). For mouse species, there was a significant difference between indoor and outdoor(x2 = 678.00, P ＜ 0.01 ). The average mouse density was 8.44%(678/8036) in trap nights. Indoor mouse density reached 14.44%(517/3581 ), which was significantly higher than that of outdoor(3.61%, 161/4455 ).For mouse density, there was a significant difference between indoor and outdoor(x2 = 301.04, P ＜ 0.01 ). When the water level was up to 156 m, mouse density reached 10%(513/5132), which was higher than that of before (5.68%, 165/2904). There was a significant difference in mouse density before and after reserving water (x2 = 44.68, P ＜ 0.01 ). With the change of water level, upstream mouse density formed a high platform from May 2007 to May 2008, followed by 12.25%(80/653), 13.16%(90/684), 12.95%(90/695), and decreased to 8.38%(28/334) after cleaning of the dam. Conclusions The Three Gorges dam and change of water level actually alter the survival environment of the local mouse, and affect local mouse density and mouse species. These may lead to local outbreak or epidemic of rodent-borne diseases.

OBJECTIVETo investigate the expression of acidic fibroblast growth factor (aFGF) in transgenic safflower and lay the foundation for the use of the plant bioreactor large-scale production aFGF.

METHODThe haFGF gene was transformed into plant preference of the aFGF sequence as a basis for design of primers, plant preferences aFGF gene sequences was amplified by PCR. The vegetable body expression vector was constructed by using digested connection method and then transferred to Agrobacterium tumefaciens EHA105 by the freeze-thaw method. It transferred to safflowers by agrobacterium-mediated transformation method, and identified by PCR, southern blot and RT-PCR.

RESULTThe full-length aFGF gene sequences were amplified through PCR and constructed into plant expression vector with soybean oleosin and promoter, and transformed into safflower. Three independently transformed safflower plant units with point insertion were successfully obtained, which showed the same size of aFGF expression at the transcriptional level.

CONCLUSIONThe plant oil body expression vectors were successfully constructed, and the optimal condition for genetic transformation was selected. The transgenic safflower plants were obtained.

Agrobacterium tumefaciens ; genetics ; Carthamus tinctorius ; genetics ; Fibroblast Growth Factor 1 ; genetics ; Gene Expression Regulation, Plant ; Genetic Vectors ; Humans ; Membrane Proteins ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; Polymerase Chain Reaction ; Transformation, Genetic

OBJECTIVETo assess the feasibility and accuracy of CT coronary angiography (CTCA) combined with adenosine stress myocardial perfusion scintigraphy (MPS) for diagnosis of flow-limiting coronary stenosis.

METHODSA total of 105 patients with suspected or established coronary artery disease (CAD) underwent CTCA and MPS within 4 weeks before invasive coronary angiography. The accuracy of CTCA/MPS in the diagnosis of flow-limiting coronary stenosis was evaluated in comparison with the results of quantitative coronary angiography and MPS.

RESULTSThe sensitivity, specificity, positive predictive value and negative predictive value of CTCA/MPS as a combined approach for detection of flow-limiting coronary stenosis were all 100%. In 16% (9/55) of the patients, revascularization procedures were performed and no flow-limiting stenosis was found.

CONCLUSIONCombination of CTCA and MPS has an excellent accuracy for detecting flow-limiting coronary stenosis as compared with quantitative coronary angiography/MPI, and can be a useful gatekeeper for revascularization procedures.

Adenosine ; Aged ; Coronary Angiography ; methods ; Coronary Stenosis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Myocardial Perfusion Imaging ; methods ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed

Morphine is a frequently used analgesic that activates the mu-opioid receptor(MOR),which has prominent side effects of tolerance.Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance,currently,there is no effective therapy to treat morphine tolerance.In the current study,we aimed to develop a monoclonal antibody(mAb)precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms.We successfully prepared a mAb targeting MOR,named 3A5C7,by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization,and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation.Treatment of two cell lines,HEK293T and SH-SY5Y,with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2(GRK2)/β-arrestin2-dependent mechanism,as demonstrated by immunofluorescence staining,flow cytometry,Western blotting,coimmunoprecipitation,and small interfering ribonucleic acid(siRNA)-based knock-down.This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR.We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid.Western blot,enzyme-linked immunosorbent assays,and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase,the in vitro biomarker of morphine tolerance,via the GRK2/β-arrestin2 pathway.Furthermore,in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alle-viated morphine tolerance in mice,and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence.Finally,intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/β-arrestin2 pathway.Collectively,our study provided a therapeutic mAb,3A5C7,targeting MOR to treat morphine tolerance,mediated by enhancing morphine-induced MOR endocytosis.The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

OBJECTIVETo investigate the clinical effects of electroacupuncture (EA) at Fenglong (ST 40) on blood lipids.

METHODSTwo hundred and four patients of hyperlipidemia were randomly divided into a Fenglong group and a Xuezhikang group, 102 cases in each group. The patients in the Fenglong group were treated with electroacupuncture at Fenglong (ST 40). After arrival of qi, the needles were connected with acupoint nerve stimulator (LH 202 H type, HANS). The primary parameters of EA: for high triglycerides (TG) type, AM 50 Hz, intensity 1 mA, needle-retained time 20 min, twice per week; for high cholesterol (CHO) type, AM 100 Hz, intensity 1 mA, needle-retained time 30 min, thrice per week; for high low-density-lipoprotein (LDL-C) type, the same parameters as the high CHO type except the tolerable and comfortable intensity; for the mixing type, corresponding methods were alternatively used. The patients in the Xuezhikang group received Xuezhikang capsule orally, 2 capsules each time and twice daily, for total 11 weeks.

RESULTSThe total effective rates of the Fenglong group and the Xuezhi-kang group were 83.0% and 85.9%, respectively, with no significant difference between the two groups (P > 0.05), and there was no significant differences in the function of regulating blood lipids between the two groups (all P > 0.05). After one month follow-up survey, the total CHO, TG and LDL-C decreased and high-density-lipoprotein (HDL-C) increased, of which there was a significant difference in TG reduction (P < 0.05). There were no relapses in both groups.

CONCLUSIONEA at Fenglong (ST 40) can effectively regulate blood lipids with a better after-effect, which can be applied as a safe and effective method to replace medication for regulating blood lipids.

Acupuncture Points ; Adult ; Aged ; Cholesterol ; blood ; Electroacupuncture ; Female ; Follow-Up Studies ; Humans ; Hyperlipidemias ; blood ; therapy ; Lipids ; blood ; Male ; Middle Aged ; Triglycerides ; blood

BACKGROUNDMonocytes and macrophages in atherosclerotic plaque lead to plaque instability. The aim of the study was to determine if plaque neovascularization led to inflammation.

METHODSPatients were consecutively enrolled if their carotid intimal media thickness was > 2 mm, as revealed by duplex ultrasound. The patients then underwent dynamic contrast enhanced magnetic resonance imaging (DCE MRI) and fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography combined with computed tomography (PET CT). A target to background ratio (TBR) of ≥ 1.25 or < 1.25 served as the cutoff point for the presence and absence of inflammation, respectively.

RESULTSTwenty-six patients underwent bilateral carotid DCE MRI and 24 patients also underwent PET CT. One hundred and fifty-five plaques were evaluated by both DCE MRI and PET CT. There was no significant difference in plaque morphology between the TBR ≥ 1.25 (n = 61) and TBR < 1.25 (n = 94) groups. No significant differences were found in plasma volume and transfer constant between the TBR ≥ 1.25 and TBR < 1.25 groups.

CONCLUSIONOur study did not find a significant correlation between plaque neovascularization and the aggregation of inflammatory cells.

Aged ; Aged, 80 and over ; Carotid Artery Diseases ; pathology ; Cell Aggregation ; Female ; Fluorodeoxyglucose F18 ; Humans ; Inflammation ; pathology ; Macrophages ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neovascularization, Pathologic ; Plaque, Atherosclerotic ; pathology ; Positron-Emission Tomography ; Tomography, X-Ray Computed