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Objective:To investigate whether coxsackievirus A 16 (CVA16) infection would affect the expression of N6-methyladenosine (m 6A) methylation-related proteins in human bronchial epithelial cells (16HBE), ICR suckling mice and SCRBA2 humanized mice and influence their subcellular localization. Methods:CVA16 was used to infect 16HBE cells at a multiplicity of infection (MOI) of 0.1 and mice at 10 7 CCID 50/ml. Changes in the expression of methyltransferases, demethylases and methylated reading proteins were analyzed by Western blot. Cellular localization of these proteins was observed using immunofluorescence. Results:The expression of m 6A methylation-related proteins was gradually reduced in CVA16-infected cells with time, but showed no obvious change in ICR suckling mice or SCRBA2 humanized mice. After infection, m 6A methylation-related proteins were redistributed in both the nucleus and cytoplasm and even degraded. Conclusions:CVA16 replication in host cells altered the expression and cellular localization of m 6A methylation-related proteins, which indicated that m 6A modification might be a new potential target for enterovirus therapy.
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This paper highlights the current research and knowledge, and reviews the incidence, pathogenesis, laboratory diagnosis andantibiotic therapy.M.genitalium is one of the major causes of nongonococcal urethritis worldwide.Its pathogenesis is not fully clear,which may be associated with toxin, immune response and persistent infection.Diagnosis is achievable only through nucleic acid amplification testing(NAAT).Azithromycin, moxifloxacin and doxycycline have been the treatments of choice for M. genitalium infections.Rapid spread of macrolide resistance as well as emergence of quinolone resistance has occurred.Increasing rates of treatment failure have resulted in an urgent need for new therapies.(Chin J Lab Med,2018,41:328-332)
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Indoleamine 2,3-dioxygenase (IDO) is a kind of immune regulation enzyme.IDO is the only rate-limiting enzyme that can catalyze the oxidative clearage of indole ring in tryptophan and its catabolism along the kynurenine pathway.IDO1 is one of the important mechanisms in maternal immune tolerance,which participates in the regulation of maternal-fetal immune relationship.The newly discovered IDO2 also plays an important role in this relationship.The abnormal expression of IDO2 is associated with the male infertility.
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We aimed to express and purify three rabies virus glycoproteins with different tags and sizes. After analyzing their binding function, we wish to obtain a rabies virus glycoprotein with higher affinity and ability to specifically bind memory B cells. Experiments were carried out to express full length, as well as the ectodomain RVG by gene engineering method. Combined with the antibody of CD19 and CD27, the candidate protein labeling with fluorescence was used to analyze its binding function. Flow cytometry was used to detect the anti-rabies virus specific memory B cells in PBMCs, and confirm the binding ability between the candidate proteins and anti-rabies virus-specific memory B cells. We successfully constructed three expression vectors pGEX-5X-1-RVG, pET28a-RVG and pET30a-G. Three glycoproteins GST-RVG, His-RVG and His-G were obtained by optimized expression and purification conditions. The antigen specificity of purified GST-RVG, His-RVG and His-G were identified by Western blotting and ELISA. The affinity of these three purified glycoproteins to anti-rabies virus antibody were detected by competitive ELISA. Anti-rabies virus specific memory B cells in positive PBMCs gained from people who had ever been injected with the vaccine can be detected by flow cytometry. Thus, we got a recombinant rabies virus glycoprotein that had high-affinity and could sort antigen specific memory B cells.
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Objective To investigate the relationship of Vaneomycin serum concentration and drug adverse reactions (ADRs) in neonates.Methods Two hundred and thirteen neonates who were treated by Vancomycin and had their serum concentration monitored were recruited.The number of cases which had liver,kidney and hearing damage after treatment was calculated.The correlation between 3 different serum concentrations (＜5 mg/L,5-10 mg/L,＞ 10 mg/L) and ADRs were analyzed.Results The mean level of Vancomycin in serum was (4.36 ± 4.99) mg/L The total incidence of ADRs was 17.8％ (38/213 cases).The main ADRs were liver damage and hearing impairment,whose incidences were 9.9％ (21/213 cases) and 7.5％ (16/213 cases),respectively,and they were were significantly higher than kidney danage (0.5％,1/213 cases) (x2 =19.172,P =0.000;x2 =13.785,P =0.000).There was no significant difference be-tween the inidence ratio of liver damage and that of the hearing impairment (x2 =0.330,P =0.566).The ADRs ratio among 3 different serum concentrations was 17.0％ (24/141 cases),17.6％ (9/51 cases) and 23.8％ (5/21 cases).There was no significant difference between the ADRs ratios and distribution of different serum concentrations (x2 =0.576,P =0.750).Moreover,the ADRs rates didn't increase along with the serum concentrations (Z =0.648,P =0.517).Except for aspartate transaminase,the indicators of liver and kidney function varied significantly after vancomycin treatment (P ＜0.05).But there was no clinical significance because the mean value was in normal range.By Bivariate Correlation analysis,Vancomyein serum concentration had no significant influence on liver and kidney function (P ＞ 0.05).Conclusions Neonates treated by Vancomycin had reatively high ratio of the liver damage and the hearing impairment.There is no significant correlation between Vancomyein serum concentration and ADRs in neonates.
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OBJECTIVE@#To evaluate the fertility of infertile males of large samples by the routine semen examination, and to provide a scientific basis for clinical treatment of male infertility and improve efficacy.@*METHODS@#We collected 16 835 semen specimens of infertile man in our hospital (the infertility group), and 1 567 semen specimens of sperm donating candidates from the sperm bank of our hospital (the control group). Semen routine examination was performed by computer-assisted semen analysis (CASA) technology, and the data of semen routine examination were analyzed statistically.@*RESULTS@#The infertile patients with semen abnormalities in our hospital mainly suffered asthenospermia. The proportion of azoospermia, asthenospermia and oligoasthenospermia in the infertility group was higher than that in the control group, while the proportion of oligozoospermia was lower than that in the control group (P<0.001).@*CONCLUSION@#Azoospermia, asthenospermia and oligoasthenospermia are related to infertility, but simply oligozoospermia is not necessarily related to infertility.
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Humans , Male , Asthenozoospermia , Azoospermia , China , Infertility, Male , Epidemiology , Oligospermia , Semen Analysis , Sperm Count , Spermatozoa , PathologyABSTRACT
OBJECTIVE@#To investigate the relationship between sperm acrosome reaction (AR) and the clinical pregnancy rate of intrauterine insemination (IUI).@*METHODS@#We detected the sperm spontaneous AR rate and Ca2+ ionophore A23187-induced AR rate in 128 patients who accepted IUI treatment, collected their clinical data and analysed the relationship between sperm AR rate and clinical pregnancy rate of IUI.@*RESULTS@#There was no statistical difference between the spontaneous AR rates in the pregnant group and the non-pregnant group (7.7% vs. 7.0%, P>0.05), but there was statistical difference between the induced AR rates(51.9 % vs. 43.5%, P<0.05). There was statistical difference in the clinical pregnancy rate among the 3 IUI groups divided by induced AR rate (≤20.0%, 20.1%-49.9%, and ≥50.0%; 4.8% vs. 12.5% vs. 18.6%, P<0.05).@*CONCLUSION@#The spontaneous AR rate has nothing to do with the clinical pregnancy rate of IUI, but the induced AR rate is associated with the clinical pregnancy rate of IUI.
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Adult , Female , Humans , Male , Pregnancy , Acrosome Reaction , Physiology , Calcimycin , Pharmacology , Infertility , Therapeutics , Insemination, Artificial, Homologous , Methods , Pregnancy Rate , Spermatozoa , PhysiologyABSTRACT
OBJECTIVE@#To determine the effect of sperm morphology on fertilization rate, cleavage rate,and clinical pregnancy rate, which may help to choose a best protocol before assisted reproductive technology (ART).@*METHODS@#We a retrospectively surveyed semen data of 1369 ART patients from January to December 2008 in our hospital. We divided sperms from 751 patients into 4 groups based on the sperm morphology rate: ≤5.0% (normal), 5.1%-10.0%, 10.1%-15.0%, and >15.0%. In vitro fertilization (IVF) was used for 490 patients or intracytoplasmic sperm injection (ICSI) for 261. We also divided sperms from 618 donors into 4 groups based on the sperm morphology rate: 14.0%-17.0%(normal), 17.1%-19.0%, 19.1%-23.0%, and >23.0%. Only IVF was used for these donors. The fertilization rate, cleavage rate, and clinical pregnancy rate among each group were analyzed and discussed.@*RESULTS@#During the IVF(patients or donors) cycle, there was a significant difference among the 4 groups in the fertilization rate and clinical pregnancy rate (P0.05). In the ICSI cycle, there was no significant difference among the 4 groups in the fertilization rate, clinical pregnancy rate, and cleavage rate (P>0.05).@*CONCLUSION@#Sperm morphology affects the outcome of IVF. There is no significant influence of sperm morphology on the outcome of ICSI. ICSI is an optimal alternative for patients with abnormal morphological sperms in ART.
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Adult , Female , Humans , Male , Pregnancy , Fertilization in Vitro , Infertility, Male , Therapeutics , Pregnancy Rate , Reproductive Techniques, Assisted , Retrospective Studies , Sperm Injections, Intracytoplasmic , Spermatozoa , Cell BiologyABSTRACT
Objective To analyze the effect of sperm factor on recurrent pregnancy lass.Methods Accordingto WHO criteria,we routinely analyzed the sperm of 21 unexplained recurrent pregnancy loss patients as study group.and 20 proven fertility men as control group.Meanwhile,we evaluated sperm chromosome aneuploidy for chromosomes 18,X and Y using standard FISH procedure,and sperm DNA ingegrity using sperm chromatin structure assay(SCSA)procedure.Results The percentage of normal morphology sperm is(10.8±2.8)%,and DFI value is(21.7±6.7)%,and the percentage of sperm aneuploidy for sex chromosome and chromosome 18 is(0.80±0.14)%and(0.23 ±0.09)%respectivelyin study group.The differences between the two groups were statistically significant(P＜0.01).Conclusion Sperm morphology,DNA integrity and chromosome aneuploidy are the correlative factors to recurrent pregnancy loss.
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<p><b>OBJECTIVE</b>To construct the two-dimensional electrophoretic (2-DE) protein map of the human sperm head.</p><p><b>METHODS</b>Protein extracts of the normal human sperm and sperm head were loaded on an 18 cm immobilized pH gradients (IPG) strip holder and separated with isoelectric focusing electrophoresis as the first dimension, and then with upright SDS-PAGE as the second. Different protein spots between them were compared with Image Master 3.0, and so were the maps of method I with Urea/Thiourea/Guanidine HCl and method II with Kit/Guanidine HCl.</p><p><b>RESULTS</b>Of the whole-sperm proteome, 802 protein spots were obtained by method I, and 797 by method II. Among them, distribution patterns of 492 spots were the same. A sperm protein map was constructed with 1107 protein spots after the interaction of the spots obtained by both the methods. Of the sperm-head proteome, 428 protein spots were obtained, all found in the whole-sperm protein map after comparison.</p><p><b>CONCLUSION</b>Both the methods could be complementarily used to construct a sperm protein map, and the map could serve as a model for the establishment of the sperm protein profile.</p>
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Humans , Male , Electrophoresis, Gel, Two-Dimensional , Methods , Hydrogen-Ion Concentration , Proteins , Proteomics , Methods , Sperm Head , ChemistryABSTRACT
OBJECTIVE:To investigate the use of antibacterials in our hospital before and after the issuing of Guiding Principles for Clinical Application of Antibacterials.METHODS:A total of 2 880 cases histories(60/month) of patients who had been discharged from hospital were randomly sampled from Oct.2002 to Sep.2006 for a review of the use of antibacterials in our hospital.RESULTS:The application rate of antibacterials before and after the issuing of the Guiding Principles for Clinical Application of Antibacterials were 94.3% and 83.8%,respectively;and the rates of combined drug use were 39.5% and 27.9%,respectively,of which,the rates of bigeminy were 30.8% and 25.3%,respectively and the rates of trigeminy were 8.7% and 2.5%,respectively.CONCLUSION:The application rate of antibacterials in our hospital was lowered significantly after the issuing of the guiding principle and the rate of antibacterials used in single kind increased while the rate of combined drug use decreased.
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OBJECTIVE: To establish an SPE(solid phase extraction)-HPLC method for the determination of diclofenac sodium in human plasma.METHODS: The separation of diclofenac sodium was performed on ODS-2 column with a detection wavelength of 280nm.The mobile phase was composed of phosphate buffer(pH6.5)-methanol(40∶ 60) at a flow rate of 1.0mL? min-1.RESULTS: The linear range of diclofenac sodium was 16.2～ 2 024.0ng? mL-1(r=0.998 9).The methodological recovery ranged between 90% to 110% and the extraction recovery was above 90%.Both intraday RSD and inter-day RSD were less than 5%.CONCLUSION: The method is simple,accurate and reliable,and suitable for the pharmacokinetic study of diclofenac sodium.
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@#Comparison of rCBF SPECT imaging and clinical visual function of 12 cases with complete homonymous hemianopia (CHH) were taken before and after a course of oriented dynamic color photic stimulation (ODCPS ). It was suggested that ODCPS in patients with CHH was an effective met hod for increasing visual field and improving visual function. Cerebral metabolic patterns of increasing rCBF reflected the mechanism of ODCPS effecting the patients with CHH. The rgtinal- midbrain-occipital visual path way may play an important role in mediating the increase of visual field and restoration of visual function.
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This article explains the purpose of setting up the sperm bank, the role of the sperm bank in auxiliary reproduction technology, and exhaustively clarifies the ethical problems in the various aspects of the social life that are caused by the sperm bank, and finally it also proposes some relevant suggestions to these problems.