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OBJECTIVE@#To construct a magnetic and catalytic hairpin assembly-based platform for detection of dual membrane proteins on exosomes.@*METHODS@#Exosomes in supernatant of breast cancer MDA-MB-231 cell culture were separated, purified and characterized. Super-resolution imaging and Western blotting were performed to confirm the expression of the membrane protein CD63 on the exosomes. Polyacrylamide gel electrophoresis was used to verify the combination of Apt-T and exosomes. Fluorescence experiments were carried out to test the feasibility of CHA nucleic acid sequence, optimize the reaction conditions, and determine the specificity of the detection platform.@*RESULTS@#Super-resolution imaging and Western blotting showed that breast cancer MDA-MB-231 cell-derived exosomes expressed abundant membrane protein CD63. Polyacrylamide gel electrophoresis indicated that Apt-T could recognize and bind to exosomes. The results of specificity test showed that the signal-to-noise ratio of the detection platform was 1.10±0.01 for detecting normal human breast epithelial cell-derived exosomes, and was 2.09±0.08 for breast cancer cell-derived exosomes.@*CONCLUSIONS@#Magnetic and catalytic hairpin assembly-based detection platform allows simultaneous detection of two membrane proteins expressed on exosomes and identification of the expressions of membrane proteins on exosomes from different sources.
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Humans , Blotting, Western , Exosomes , Magnetic Phenomena , Membrane Proteins , Sensitivity and SpecificityABSTRACT
Objective To investigate the changes of coagulation function in patients with sepsis after anti-coagulant treatment and its correlation with prognosis.Methods The clinical data of 132 patients with sepsis in our hospital were analyzed retrospectively,and all the patients were given routine anti-infection,fluid re-placement,acid suppression,organ support and anticoagulant treatment.The prognosis was recorded within 1 month after admission,the patients enrolled in the study were divided into the death group(46 cases)and the survival group(86 cases).T he sex,age and coagulation function[prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fibrinogen(FIB),two D-dimer(D-D),platelet count (PLT)]of the patients were compared between the two groups.Logistic multiple factor regression analysis was performed.At the same time,ROC curves for predicting the risk of death in patients by using coagulation function were drew.Results After the anticoagulant treatment,PT,APTT,TT,FIB,D-D values of the sur-vival group were lower than the death group,and PLT was higher than the death group(P < 0.05).Logistic multivariate regression analysis showed that PT,APTT and PLT were independent factors in the mortality of sepsis patients(P<0.05).The area under the curve(AUC)of PT for predicting death risk was 0.815,when the best cut-off value was 13.84 s,the sensitivity was 0.870 and specificity was 0.698.The AUC of APTT for predicting death risk was 0.756.,when the best cut-off value was 38.95 s,the sensitivity was 0.848 and speci-ficity was 0.698.The AUC of PLT for predicting death risk was 0.826,when the best cutoff value was 216.62×109,the sensitivity was 0.686 and the specificity was 0.913.Conclusion After anticoagulant treat-ment ,PT ,APTT and PLT are independent prognostic factors of sepsis ,and they have some predictive valuefor the risk of death .
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Objective: To explore risk factors for complicated cardiovascular and cerebrovascular diseases in patients with refractory hypertension (RH).Methods: Clinical data of 536 RH patients, who were treated in our hospital from Jun 2010 to Jul 2015, were retrospectively analyzed.According to presence of cardiovascular and cerebrovascular diseases or not, patients were divided into complication group (n=217) and pure RH group (n=319).Results: Compared with pure RH group, there were significant rise in percentages of drinking and smoking, hypertension course, mean arterial pressure (MAP), mean systolic blood pressure, pulse pressure, serum uric acid (SUA), creatinine (Cr), total cholesterol and low density lipoprotein cholesterol (LDL-C), and significant reductions in high density lipoprotein cholesterol (HDL-C) mean diastolic blood pressure and mean heart rate in complication group, P<0.01 all.Multi-factor unconditional Logistic regression analysis indicated that hypertension course, SUA,MAP, Cr and LDL-C levels were independent risk factors for complicated cardiovascular and cerebrovascular diseases in RH patients (OR=1.014~3.217, P<0.05 or <0.01).Conclusion: Hypertension course,MAP,Cr,SUA and LDL-C are independent risk factors for refractory hypertension.Enhancing their monitor, prevention and treatment are important to reduce incidence rates of cardiovascular and cerebrovascular complications in patients with refractory hypertension.
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Objective To establish a rabbit model of facial skin lifting and to evaluate the skin lifting effects of absorbable soft tissue plastic belt, and to provide experimental basis for clinical application of this plastic belt.Methods 36 Japanese white rabbits were randomly divided into model control group (n=6) and plastic belt lifting group (n=30). The model control group received only facial skin resection surgery, while the plastic belt lifting group had facial skin resection and plastic belt implantation.The changes of rabbit general status, skin lifting effects, performance of plastic belt and pathological changes of subcutaneous tissue after implantation were determined during the 4 weeks after surgery. Results The rabbit skin wound was healed within 3-7 days after surgery without infection.Meanwhile, the plastic belts did not show shifting or rupture, and only whitening around the perforations was observed in the two groups.Compared with the model control group, the MA and MB distances in the plastic belt lifting group were significantly lower ( P0.05 ) .Moreover, the molecular weight ( Mw ) , peak molecular weight ( Mp ) , Z molecular weight (Mz) and viscosity were gradually decreased along with the time passing after implantation (P<0.01), and its dispersion Mz/Mw ratio was also gradually decreased from 2 weeks after implantation (P<0.01), and no obvious pathological changes were found after subcutaneous implantation of the plastic belts.Conclusions We have successfully established a facial skin lifting rabbit model, and the plastic belt can obviously lift the facial skin fascia system and keep intact more than 4 weeks in the body.Therefore, this plastic belt can be applied for anti-wrinkle facial soft tissue lifting against the skin damage caused by aging.
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Objective To investigate the effect of the replication-deficient adenovirus carrying miR-138(Ad-miR138) on the pro-liferation of human gastric cancer cell line BGC-823 and the possible mechanism .Methods The human gastric cancer cell line BGC-823 was infected with Ad-miR138 .Then the expression level of miR-138 was measured by RT-PCR .The growth curve of BGC-823 was measured using CCK-8 method .The ability of cell invasion was measured using Transwell chambers .Results After infected with Ad-miR138 ,the expression of miR-138 in BGC-823 cells was up -regulated significantly (1 .07 ± 0 .07 vs .4 .89 ± 0 .45 ,P<0 .05) .Absorbance of the 6th day decreased significantly (0 .52 ± 0 .06 vs .0 .77 ± 0 .06 ,P<0 .05) ,and the invasion ability was de-creased obviously (32 .00 ± 11 .00 vs .56 .00 ± 12 .00 ,P<0 .05) .Conclusion miR-138 can effectively suppress the proliferation and invasion of human gastric cancer cell line BGC-823 in vitro .
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Objective To provide an experimental reference for clinic application of Jiazhu vascular ligating clips on ligation effects and the impact on the body in rabbits after laparoscopoic cholecystectomy.Methods 36 rabbits were randomly divided into 3 groups, normal control group, Jiazhu group and similar product control ( Hem-o-lok) group, 12 rabbits in each group.Except for normal control group, the rabbits were used gallbladder duct and gallbladder artery closed by vascular ligating clips and Hem-o-lok clips in Jiazhu group and similar product control group, respectively.The changes of blood biochemical, electrolytes, hematological parameters, coagulation, ligation effects and organ coefficients were observed at post-operation during 12 months.Results Compared with normal control group, the rabbit weight was reduced in the Jiazhu group at 3 days postoperatively ( P 0.05 ) , and there were all no significant differences with electrolyte, coagulation and organ coefficients between each group (P >0.05).Conclusion Jiazhu vascular ligating clips are safe and reliable to close gallbladder duct and blood vessels, It shows that there are no differences in the validity and security between Jiazhu vascular ligating clips and Hem-o-lok clips.
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OBJECTIVE@#To evaluate the impact of sperm source and sperm parameters on the outcome of intracytoplasmic sperm injection (ICSI).@*METHODS@#This retrospective study included 433 ICSI cycles from June 2005 to December 2008 in Reproductive Medical Center of Xiangya Hospital. The patients were divided into 2 major groups according to the source of spermatozoa used for ICSI: ejaculated (group A, n=336) and epididymal (group B, n=97). Group A was divided into 3 subgroups according to the sperm parameters: normal (Group A1, n=95), single parameter defect (Group A2, n=119), and multiple parameter defect (Group A3, n=122).@*RESULTS@#The basic characteristics among the 4 groups had no statistic difference (P>0.05), and the difference in the fertilization rate, normal fertilization rate, cleaving embryo rate,good quality embryo rate, implanted rate, clinical pregnancy rate, and early abortion rate among the 4 groups were not significant (P>0.05).@*CONCLUSION@#The outcome is similar no matter whether the spermatozoa is from ejaculated sperm or epididymis. ICSI can treat male infertility of various factors, and the outcome is the same with one or multiple sperm parameter abnormality. ICSI with epididymal spermatozoa through percutaneous epididymal sperm aspiration is effective for infertility due to obstructive azoospermia.
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Adult , Female , Humans , Male , Pregnancy , Epididymis , Cell Biology , Fertilization , Infertility, Male , Therapeutics , Pregnancy Outcome , Retrospective Studies , Sperm Injections, Intracytoplasmic , Methods , Sperm Motility , Physiology , Sperm Retrieval , Spermatozoa , PhysiologyABSTRACT
OBJECTIVE@#To determine the sperm-zona pellucida (ZP) binding and ZP-induced acrosome reaction in patients with unexplained infertility, and to discuss the relationship between ZP-induced acrosome reaction and fertilization rate.@*METHODS@#We compared the fertilization rate and good embryo rate in patients with unexplained infertility after fertilization in 2 ways. Based on the causes of infertility, patients were divided into an unexplained infertility group (Group A) and a pure female tubal factor group (Group B). Oocytes which were obtained by super ovulation from 25 patients with unexplained infertility were randomly divided into 2 groups with conventional in vitro fertilization (IVF) (Group A1) and intracytoplasmic sperm injection (ICSI) fertilization (Group A2). The pure female tubal factor group (Group B) had conventional IVF. We conducted sperm-ZP binding and ZP-induced acrosome reaction experiments with 2 groups of men's sperms separately. We compared the number of sperm-egg binding and ZP-induced acrosome reaction rate and discussed the relationship between the ZP-induced acrosome reaction and fertilization rate, and also the fertilization rate, good embryo rate and pregnancy rate in patients with unexplained infertility after fertilization in 2 ways.@*RESULTS@#The average number of sperm-egg binding (78.29 ± 16.31) and the ZP-induced acrosome reaction rate (55.87 ± 27.69) % in Group A were lower than those of Group B [94.63 ± 6.72, (82.53 ± 17.99)%]. The difference between the average number of sperm-egg binding and the ZP-induced acrosome reaction was significant (P <0.01). The fertilization rate of Group A1 was significantly lower than that of Group B and Group A2 (P <0.01). But there was no significant difference in the good embryo rate among the 3 groups. There was no significant difference between Group A2 and B in fertilization rate and good embryo rate (P <0.05). There was no significant difference in pregnancy rate between Group A and B (P <0.05). Fertilization rate and the rate of acrosome reaction had marked positive correlation with statistical significance (r =0.932, P <0.01).@*CONCLUSION@#ZP binding and ZP-induced acrosome reaction are very important experiments in sperm function test for patients with unexplained infertility. It can not only effectively avoid no embryo transferring due to complete failure of fertilization but also get a desirable outcome of pregnancy using half-ICSI fertilization in patients with unexplained infertility.
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Female , Humans , Male , Acrosome Reaction , Embryo Transfer , Fertilization in Vitro , Infertility , Therapeutics , Oocytes , Physiology , Ovulation Induction , Sperm Injections, Intracytoplasmic , Sperm-Ovum Interactions , Physiology , Treatment Outcome , Zona Pellucida , PhysiologyABSTRACT
OBJECTIVE@#To explore the different effect of short 7-day gonadotropin releasing hormone agonist (GnRHa) protocol and GnRHa long protocol on the insulin-like growth factor II(IGF-II) and insulin-like growth factor binding protein-4 (IGFBP-4) levels in follicular fluid.@*METHODS@#Eighty-eight infertile patients due to tubal factors were included in this study. They were randomly divided into a short 7-day GnRHa protocol group and a GnRHa long protocol group (n = 44). Follicular fluid was obtained from dominant follicles during oocyte retrieval. Levels of IGF-II and IGFBP-4 in the follicular fluid were detected by radioimmunoassay and enzyme-linked immunosorbent assay respectively.@*RESULTS@#Duration of controlled ovarian stimulation was significantly shorter and the injected dosages of gonadotropin were significantly lower in the short 7-day protocol group. The differences in serum levels of estradiol and estradiol per mature follicle on the day of human chorionic gonadotropin injection between the two groups were not significant. The concentrations of IGF-II and IGFBP-4 in the follicular fluid of the short 7-day protocol group were significantly lower,while the difference of the ratio of IGF-II/IGFBP-4 between the two groups was not significant. Linear correlation analysis showed that IGF-II level in the follicular fluid was positively correlated to the total dose of gonadotropin.@*CONCLUSION@#The short 7-day and long GnRHa protocols may affect the concentrations of IGF-II and IGFBP-4 in the follicular fluid. However, changes of IGF-II and IGFBP-4 concentrations do not contribute to different clinical outcomes.
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Female , Humans , Embryo Transfer , Fertilization in Vitro , Methods , Follicular Fluid , Chemistry , Gonadotropin-Releasing Hormone , Insulin-Like Growth Factor Binding Protein 4 , Insulin-Like Growth Factor II , Ovulation Induction , MethodsABSTRACT
SR proteins play important roles in regulating alternative pre-mRNA splicing. As a newly discovered neural and reproductive tissue specific SR protein, SRp38 regulates the alternative splicing of several genes important for neural function, such as GluR-B, Trk-C and NCAML1. It also acts as a splicing inhibitor during mitosis or stress response in order to prevent wrong splicing. The expression of SRp38 in mouse retina was investigated by Western blot and immunohistochemistry (IHC) analyses. The result shows that the expression of SRp38 proteins in mouse retina is region-specific, with extensive distribution in the outer and inner plexiform layers, inner nuclear layer and ganglion cell layer, but no expression in outer nuclear layer. Double staining of isolated retina cells with anti-SRp38 and anti-Trk-C antibodies showed that SRp38 is localized in the dendrites, somata and axon terminals of rod-bipolar cells. By transient co-transmission of over-expressed SRp38 plasmid and RT-PCR analyses, the further results showed that overexpressed SRp38 could promote the splicing of the Flip isoform of GluR-B minegene in R28 cells. The result suggests that SRp38 may play important roles in the retinal function, possibly via regulating the neural-specific alternative splicing of genes as GluR-B.
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Gliomas constitute the most common type of primary brain tumor. Alternative pre-mRNA splicing may play roles in the carcinogenesis, cell growth and invasion in various types of gliomas. Factors regulating the alternative spicing in gliomas include cis-regulatory element (for example, ESE, ISS and ESS) and trans-regulatory factors (such as SRp55, SC35, SF2/ASF and PTB). Recent progresses demonstrate that many genes associated with gliomas, including those encoding tumor suppressors or promoters, enzymes, receptors and ion channels are subject to regulation by alternative pre-mRNA splicing. Therefore, studying the alternative pre-mRNA splicing in gliomas will be of benefit to understanding the molecular basis of gliomas and identifying new targets potentially useful for early diagnose as well as therapy of gliomas.
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Objective To discuss the causes of syncope and its influencing factors for medical students during the comprehensive military training in military medical university and explore the measures to prevent the incidence.Methods Combined with practical exercises,the clinical data of 42 cases of syncope during the comprehensive military training were retrospectively investigated and analyzed.Results During the military training,the percentage of vascular dysfunction syncope was 78.87%,and girls had higher rate than boys(Chi square=28.27,P
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Objective:To prepare specific antibody against the pre mRNA splicing protein Tra2? in human fetal tissues.Methods:A specific antigen fragment of human Tra2? was cloned into the pGEX 3x expression vector to express a recombinant GST Tra2? fusion protein.The fusion protein was expressed,purified and used as an antigen to immunize the New Zealand rabbit,and the polyclonal antibody to Tra2? was obtained.Results:The antibody was successfully used to detect Tra2? protein in human fetal cerebrum with Western blot and immunohistochemistry analysis.Conclusion:This prepard antibody has good specificity and can be used to detect the Tra2? protein in the human fetal tissues.