الملخص
Antisperm antibodies (ASAs) are assumed to be a possible causative factor for male infertility, with ASAs detected in 5%-15% of infertile men but in only 1%-2% of fertile ones. It remains unclear whether ASAs have an adverse effect on the outcome of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). This study investigated differences in the rates of fertilization, pregnancy, and live births associated with serum ASA-positive and ASA-negative men following IVF or ICSI. Five hundred and fifty-four consecutive infertile couples undergoing IVF (n = 399) or ICSI (n = 155) were included. The two-sample two-sided t-test and Chi-square or Fisher's exact test was used for statistical analysis. Lower rates of fertilization (41.7% vs 54.8%, P = 0.03), good embryos (18.9% vs 35.2%, P = 0.00), pregnancy (38.5% vs 59.4%, P = 0.00), and live births (25.8% vs 42.5%, P = 0.00) were observed in men of the IVF group with a positive serum ASA than in those with a negative ASA. ASA positivity/negativity correlated with pregnancy rates (P = 0.021, odds ratio [OR]: 0.630, 95% confidence interval [CI]: 0.425-0.932) and live birth rates (P = 0.010, OR: 1.409, 95% CI: 1.084-1.831) after controlling for the female serum follicle-stimulating hormone level and the couple's ages at IVF. Women coupled with ASA-positive men had lower live birth rates with IVF than with ICSI (25.8% and 47.4%, respectively; P = 0.07). Women coupled with ASA-positive men had lower rates of pregnancy and live births following IVF than those coupled with ASA-negative men but had a similar outcome with ICSI.
الموضوعات
Adult , Female , Humans , Male , Pregnancy , Young Adult , Antibodies/pharmacology , Cohort Studies , Fertilization , Fertilization in Vitro/methods , Infertility, Male/therapy , Live Birth , Pregnancy Outcome , Pregnancy Rate , Sperm Injections, Intracytoplasmic/methods , Spermatozoa/immunology , Treatment Outcomeالملخص
Aim To explore the influence of mechani-cal environment on differentiation of mesenchymal stem cells ( MSCs) at fracture site during early stage of frac-ture healing. Methods The rat femur fracture models were established and fixed with external fixation frame of different stiffness coefficient and were divided into three groups, namely, the sham group with super-high stiffness external fixation frame, the high stiffness fixa-tion frame group and the low stiffness fixation frame group. The bone callus and the degree of fracture heal-ing after two weeks compared with the very day after surgery were observed by X-ray. And the effects were evaluated respectively by histopathology 6 weeks after operation. Callus tissues of 2 days, 6 days, 10 days, 14 days after operation were acquired. Runx2, Osterix and Sox9 expression were measured by RT-qPCR. Runx2 and Sox9 were tested by Western blot. Results X-ray showed that the amount of callus of high stiff-ness group was significantly less than that of low stiff-ness group. Histological study showed, sham group and high stiffness group achieved less callus and had lots of active bone cells while low stiffness group got cartilage ossification well. The results of RT-qPCR showed that the level of Runx2 , Osterix increased with-in 14 days. And Sox9 reached the highest level in sham group and high stiffness group 10 days after sur-gery and declined in 14 days while low stiffness group had been rising within 14 days after surgery. The ex-pression levels of Runx2 , OSX and Sox9 in 14 days af-ter operation presented statistically significant differ-ence among the three groups at the same time point, i. e. low stiffness group > high stiffness group > the sham group ( P <0.05 ) . The results of Western blot were consistent with those of RT-qPCR. Conclusion The differentiation of MSCs at fracture site is affected by the mechanical environment, and subtle movement could promote the healing of fracture sites.
الملخص
<p><b>OBJECTIVE</b>To investigate the relationship between the promoter polymorphism of IL-4 and IL-6 and chronic rhinosinusitis (CRS).</p><p><b>METHODS</b>One hundred and twenty-three patients with CRS and 239 healthy controls in Shanghai region were chosen in this study. The genotype of IL-4 gene -33T>C and -590C>T were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and the genotype of IL-10 gene -1082A>G was determined using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method. Statistical calculations were performed using SAS 8.2 software.</p><p><b>RESULTS</b>Significant differences were found in genotype distribution of -33T>C and -590C>T between the CRS group and the control group (χ2=6.6013, P=0.0102, χ2=6.6013, P=0.0304), and -33T>C remained significant following application of the Bonferroni correction (P<0.025). The relative risks of CRS with -33T>C and -590C>T were 1.818(P=0.0236, 95%CI 1.084-3.050) and 1.838 (P=0.0147, 95%CI 1.127-2.997). There was linkage disequilibrium (LD) between the -33T>C and -590C>T. The coefficient of linkage disequilibrium (D') was 0.77 and the related coefficient (r2) was 0.54. The -33T/-590T haplotype was associated with CRS and the relative risk was 1.653 (P=0.0130, 95%CI 1.107-2.469). There were only two genotypes of IL-10 gene-1082A>G and the frequencies of the AA and AG genotypes were not different between the CRS and control groups.</p><p><b>CONCLUSION</b>The promoter polymorphism of IL-4 -33T>C and -590C>T were associated with the susceptibility of CRS and the -33T/-590T haplotype was a risk factor for CRS, but there were no association between the -1082A>G and CRS.</p>
الموضوعات
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Alleles , Case-Control Studies , Chronic Disease , Genetic Predisposition to Disease , Genotype , Interleukin-10 , Genetics , Interleukin-4 , Genetics , Nasal Polyps , Genetics , Polymorphism, Single Nucleotide , Sinusitis , Geneticsالملخص
<p><b>OBJECTIVE</b>To investigate the effects of knockdown of Aurora-A by RNA interference on laryngeal cancer Hep-2 cell growth in vitro and in vivo.</p><p><b>METHODS</b>A plasmid containing siRNA against Aurora-A was constructed and transfected into human laryngeal cancer cell line Hep-2. Measurements included the CCK-8 assay for viability and proliferation, Transwell assay for invasion, colony formation assay for cell anchorage-independent growth. Western blot and immunohistochemistry assay for protein expression. Tumorigenicity was observed in vivo.</p><p><b>RESULTS</b>In Hep-2 cells transfected by Aurora-A siRNA (designated as siRNA-3), protein expression of Aurora-A was suppressed by 52%. In CCK-8 assay, absorbance value of siRNA-3 cells (3.268 ± 0.106, (x(-) ± s)) was lower than that of Hep-2 cells (3.722 ± 0.152, F = 17.634, P < 0.001). In Transwell assay, the average invasive cells per field in siRNA-3 cells (110.0 ± 18.0) was less than that in Hep-2 cells (236.0 ± 26.0, F = 26.462, P < 0.01). In colony formation assay, the average colony number of siRNA-3 cells (31.0 ± 6.6) was lower than that of Hep-2 cells (104.0 ± 14.0). The average tumor size in siRNA-3 group was (127.77 ± 174.83) mm(3), which was less than Hep-2 cell group (837.26 ± 101.80) mm(3), (F = 28.187, P < 0.001). Silencing of Aurora-A decreased the expression of focal adhesion kinase (FAK) and matrix metalloproteinase-2 (MMP-2), key regulators in cell adhesion and invasion.</p><p><b>CONCLUSIONS</b>The knockdown of Aurora-A inhibits the growth and invasiveness of Hep-2 cells in vitro and in vivo, which may be a promising therapeutic strategy for LSCC.</p>
الموضوعات
Animals , Humans , Mice , Aurora Kinase A , Aurora Kinases , Carcinoma, Squamous Cell , Genetics , Metabolism , Pathology , Cell Line, Tumor , Cell Transformation, Neoplastic , Focal Adhesion Protein-Tyrosine Kinases , Metabolism , Gene Silencing , Laryngeal Neoplasms , Genetics , Metabolism , Pathology , Matrix Metalloproteinase 2 , Metabolism , Mice, Nude , Protein Serine-Threonine Kinases , Genetics , RNA Interference , RNA, Small Interfering , Genetics , Transfectionالملخص
Objective To construct a novel enhanced green fluorescent protein (EGFP) and glial cell line-derived neurotrophic factor (GDNF) recombinant baculovirus. Methods The target gene(EGFP and GDNF) was cloned into baculovirus transfer vector pFastBacDual, pFB-EGFP-GDNF was constructed and restriction enzyme analysis was conducted. pFB-EGFP-GDNF was transposited with baculovirus shuttle vector (Bacmid) into DH10Bac competent cells, and recombination baculovirus vector Bacmid-EGFP-GDNF was constructed. The plasmid was extracted and PCR was performed for identification. Bacmid-EGFP-GDNF was transfected with Sf9 insect cell package virus by liposomal transfection method. Immunofluorescent staining was employed to detect the expression of EGFP and GDNF protein in St9 cells. Results The target gene fragment was correctly cloned into pFastBaeDual vector, and recombinant Bacmid was constructed. Bacmid-EGFP-GDNF was successfully transfected, and higher virus titer was obtained. The coexpression of GDNF and EGFP protein in Sf9 cells was identified by immunofluorescent staining. Conclusion The recombinant baculovirus Bacmid-EGFP-GDNF can be successfully constructed, and the protein of EGFP and GDNF is coexpressed in St9 cells, which paves a way for the research of GDNF gene therapy.
الملخص
<p><b>OBJECTIVE</b>To study the feasibility and the characteristics of recombinant baculovirus as spiral ganglion cells (SGC) gene transfer vector.</p><p><b>METHODS</b>After the generation of baculovirus- green fluorescent protein( Bac-GFP) according to Bac-to-Bac baculovirus expression system, SGC were infected by Bac-GFP with different multiplicities of infection (MOI) and different concentrations of sodium butyrate. The transfection cell rate and mean fluorescence strength (MFS) were detected by fluorescence microscopy and flow cytometry. Toxicity effects of recombinant baculovirus vectors and sodium butyrate on SGC were determined by spectroscopic measurement of 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-diphenytetrazoliumromide (MTF).</p><p><b>RESULTS</b>Baculovirus was able to infect primary SGC cultures. The dose-response characteristics of Bac-GFP were determined on SGC, and the expression level could be up-regulated by sodium butyrate. Infection with Bac-GFP in the absence or presence of sodium butyrate (< or =10 mmol/L) was considered to be non-cytotoxic to primary SGC. GFP had been expressed in SGC at 6 h post-infection and the highest numbers of cells expressing GFP were observed at approximately 48 h post-infection.</p><p><b>CONCLUSIONS</b>Baculovirus is a novel and promising tool for gene transferring into the cochlear nervous system both for studies of the function of foreign genes and the development of gene therapy strategies.</p>
الموضوعات
Animals , Rats , Baculoviridae , Genetics , Cells, Cultured , Gene Transfer Techniques , Genetic Vectors , Green Fluorescent Proteins , Genetics , Rats, Sprague-Dawley , Spiral Ganglion , Cell Biology , Transfectionالملخص
Objective To explore the clinical features,diagnosis and treatment of serious pyogenic infection in deep neck. MethodsFrom January 2002 to April 2005,the clinical data of 22 patients hospitalized with serious pyogenic infection in deep neck were collected and retrospectively analysed. Results All the patients suffered unilaterally.Besides the symptoms of infection,dyspnea,pain in swallowing,limitation of mouth opening and progressive limitation of cervical activity were presented.Bacterial cultivation were performed in 14 patients and positive results were detected in six.Fourteen of all the 22 patients were cured with antibiotics,and abscess incision drainage was carried out in the other eight.Preoperative cervical PA-LAT X-ray photography and contrast-enhanced CT scanning were performed on one and seven patients,respectively,all of which indicated the formation of abscess.All the patients were cured except one who died of cerebral infarction and multiorgan dysfunction. Conclusion Imaging evaluation plays a significant role in the preoperative diagnosis and systemic treatment of serious pyogenic infection in deep neck.Bacterial cultivation provides reliable evidence for the etiopathogenisis and helps to make the effective treatment.
الملخص
Allergic rhinitis and rhinosinusitis often complicate with asthma,and their relationship has long been investigated.From the view of epidemiology,all of these three diseases have higher prevalence,complicate with each other,are risk factors and prognostic factors for each other.Besides,they share common in anatomy and pathophysiology.In this paper,the interrelationship among allergic rhinitis,rhinosinusitis and asthma in pathogenesis is discussed.
الملخص
Muscle recently has been identified as a good source of adult stem cells that can differentiate into cells of different lineages.Researchers have identified two types of stem cells in skeletal muscle.Further research is necessary to delineate the relationship between different populations of musclederived stem cells(MDSCs)and between MDSCs and other adult stem cells.The methods used to isolate these cells appear to influence the stem cell characteristics.As these efforts continue,the potential for MDSCsbased therapy for other musculoskeletal injuries,as well as for cardiac and smooth muscle injuries,is currently being explored.The behavior,biocharacteristic,isolation,differentiation and the probability of application to regenerate lost or diseased tissue of MDSCs were summarized.
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Objective To observe the changes of the concentration of Ca2+, contents of cAMP, CaM and activity of Ca2+/CaM-PK II in pheochromocytoma PC12 cells after combined soman and hypoxia injury. Methods The changes of [Ca2+], and activity of CaM, cAMP and Ca2+/CaM-PK II in PC12 cells were studied after combined soman and hypoxia injury with radioimmunoassay. Results The changes of [Ca2+], the contents of CaM, cAMP were significantly higher in hypoxic and soman intoxicated group than in soman intoxicated group and control group under hypoxia; but the activity of Ca2+/CaM-PK Ⅱ were significantly decreased. Conclusion [Ca2+], CaM, cAMP and Ca2+/CaM-PK Ⅱ exert important role in the damage of PC12 after combined soman and hypoxia injury.
الملخص
Objective To observe the changes of the concentration of Ca2+, contents of cAMP, CaM and activity of Ca2+/CaM-PK II in pheochromocytoma PC12 cells after combined soman and hypoxia injury. Methods The changes of [Ca2+], and activity of CaM, cAMP and Ca2+/CaM-PK II in PC12 cells were studied after combined soman and hypoxia injury with radioimmunoassay. Results The changes of [Ca2+], the contents of CaM, cAMP were significantly higher in hypoxic and soman intoxicated group than in soman intoxicated group and control group under hypoxia; but the activity of Ca2+/CaM-PK Ⅱ were significantly decreased. Conclusion [Ca2+], CaM, cAMP and Ca2+/CaM-PK Ⅱ exert important role in the damage of PC12 after combined soman and hypoxia injury.