Objective: To analyze the influence of semen processing methods on the proportion of the aneuploid sperm by detecting the sperm's chromosome X, Y, 18 using fluorescence in situ hybridization. Methods: Ten patients with mild ohgoasthenosperia, who were received ICSI treatment, were included in this study. Five semen samples of the patients were randomly selected to detect using Swim-up method (A group) and 5 using sperm-grad double-density centrifugation method (B group).Another 5 patients with mild oligo-asthenosperia were as control (C group). The CEPX / Y and CEP18 probe was used to detect the sperm of these 15 patients by fluorescence in situ hybridization. The proportion of aneuploid sperm was compared in three groups. Results: The sex chromosome aneuploid rates were (4.21±2.49)%, (3.24～1.49) % and (2.62±0.89) % in control, A and B groups. The rates of aneuploid chromosome 18 were (3.00±1.22)%, (2.00～1.22)% and (2.00±1.22)% in control, A and B groups. There were no significant differences in three groups (P＞0.05). Conclusion: The results showed that the methods of Swim-up and Sperm-Grad double-density gradient centrifugation could select sperms in motility potential and teratospermia,but not in normal chromosome sperms.

Objective To assess the variations in different thyroid stimulating hormone(TSH) and free thyroxine (FT4) detection kits for evaluating thyroid function during pregnancy and to establish the corresponding normal reference ranges.Methods This study was based at the International Peace Maternity and Child Health Hospital affiliated to Shanghai Jiaotong University School of Medicine.A total of 200 pregnant women who visited the hospital between June,2011 and September,2012 were recruited in this study according to the National Academy of Clinical Biochemistry (NACB) criteria.Blood samples were sequentially collected from the women at the first (T1,9-12 weeks),second (T2,16-24 weeks) and third (T3,32-36 weeks) trimesters to determine the serum TSH and FT4 levels using four different detection kits (Siemens-C,Siemens-Ⅰ,Abott and Roche).A linear trend test was used to analyze serum TSH and FT4 levels with four different kits.A percentile range of P2.5 to P97.5 was used to establish the normal trimester-dependent reference ranges of TSH and FT4 levels for different detection kits.The Bootstrap method was used to compare the differences in the four reference ranges.Results Similar dynamic changes in TSH and FT4 levels during pregnancy were detected among the different kits (F=0.950,P=0.595; F=11.640,P=0.081,respectively).Among the four reference ranges of TSH,the Roche kit showed the most remarkable fluctuation during pregnancy,while Roche kit in the first trimester and Siemens C kit in the second and third trimesters showed larger fluctuations in reference ranges of FT4.More importantly,the reference ranges of TSH and FT4 showed significant variations among the four different kits in each trimester (TSH:T1:F=2 945.390,P ＜ 0.01; T2:F=2 826.260,P ＜ 0.01; T3:F=1 698.360,P ＜ 0.01.FT4:Tl:F=1 145.440,P ＜ 0.01; T2:F=2 260.240,P ＜ 0.01; T3:F=1 439.920,P ＜ 0.01).Conclusions TSH and FT4 measurement using four different commercial kits showed similar trimester-dependent dynamic changes.However,it is necessary to establish trimester-dependent and detection kit dependent normal reference ranges of TSH and FT4 for thyroid function evaluation for pregnant women.

Objective To investigate the appropriate time of replacing drainage bags after trans-cervical resection of adhesion(TCRA) surgery.Methods Totally 156 patients underwent TCRA were randomly divided into three groups:Group A,the drainage bags were replaced daily;Group B,the drainage bags were replaced every 3 days;Group C,the drainage bags were never replaced.The drainage bags were removed on the 6th day after TCRA for all groups.Bacterial culture results from the balloon surface of drainage bag,outer cervical orifice,and the connector of drainage bag for 3 groups were observed.The positive rates of bacterial cultures were analyzed statistically.Results The bacterial culture results from the drainage bag connector of 3 groups indicated that the positive rate of Group C was significantly lower than that of other two groups(P＜0.05).The bacterial culture from the drainage bag balloon surface also indicated the same result,but the result did not showed statistically significant difference(P＞0.05).There was no statistically significant difference in the culture result firom outer cervical orifice and the volume of drainage fluid within three groups(P＞0.05).Conclusion It is indicated that the patient may enjoy a lower rate of bacterial infection without replacing any drainage bags connected to balloon drainage tubes placed after TCRA within 6 days.We suggest that the intrauterine drainage hags may be kept without replacement until direct removing the drainage tube on day 6 after TCRA.

Objective: To observe the protective effects of the alcohol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceuson on H2O2-induced oxidative damage of vascular endothelial cells ECV-304.Methods: ECV-304 cells were cultured in vitro and H2O2-induced oxidative damage in ECV-304 cells was used as the model.After the treatment with ethanol extract, ethyl acetate extract or 1-butanol extract of Euonymus hederaceus (the final concentration of 400, 200, 100, 50 and 25 μg·ml-1) for 24h, the cell viability was detected by MTT assay.Hoechst staining was used to observe the cell apoptosis, and then the supernatant was collected, and the contents of SOD, NOS and NO were used to determine the effects of the extracts on the oxidative stress of vascular endothelial cells.Results: The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus within the concentration range of 25-400 μg·ml-1 showed no cytotoxicity on ECV-304 cells, and the ethyl acetate extract at the concentration of 400μg·ml-1 had certain proliferation effect on the cells (P<0.05).The ethyl acetate extract at the concentration of 50-400 μg·ml-1 and the 1-butanol extract at the concentration of 100-400 μg·ml-1of Euonymus hederaceus could increase the survival rate of ECV-304 cells with oxidative injury significantly, and the OD value showed significant difference when compared with that of the model group (P<0.05 or P<0.01).The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could significantly improve the SOD and NO levels in ECV-304 cells with oxidative damage (P<0.05 or P<0.01), and had no notable influence on NOS level.The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could inhibit the cell apoptosis with oxidative damage and improve the nuclear morphology, and the apoptosis exhibited significant difference when compared with that of the model group (P<0.01).Conclusion: The extracts of Euonymus hederaceus have protective effects on vascular endothelial cells with oxidative damage induced by H2O2.

BACKGROUND:A few of studies have showed similar efficacy and safety between domestic clopidogrel (Talcom?) and imported clopidogrel (Plavix?) in patients after percutaneous coronary intervention, but there is lack of large-scale, large sample, and prospective clinical comparative study in China.
OBJECTIVE:To compare the efficacy and safety of Talcom?and Plavix?on percutaneous coronary intervention.
METHODS:Total y 1 798 patients with Coronary atherosclerotic heart disease who received percutaneous coronary interventions were divided to two groups:Talcom?group (n=1 104) and Plavix?group (n=694). 300 mg clopidogrel was administrated oral y before percutaneous coronary intervention fol owed by 75 mg/d clopidogrel sequential y for 1 year. Al the patients were fol owed for 3-28 months to observe the incidence rate of stent thrombosis at acute, subacute, late, and very late stage, major adverse cardiac events of combination end point (including myocardial infarction, cardiac death, and stroke), and correlated adverse reactions, such as bleeding.
RESULTS AND CONCLUSION:There were no significant differences in the incidence of stent thrombosis, target vessel revascularization, cardiac death, bleeding, major bleeding and major adverse cardiac events of combination end point between Talcom?group and Plavix?group. In addition, event-free survival also showed no difference between the two groups. After treatment, white blood cellcount, erythrocyte count, hemoglobin, hematocrit, platelet count were significantly decreased in both the two groups (P<0.05). Hemoglobin level in the Talcom?group was fewer than that in the Plavix?group (P<0.05). The results suggest that effects and safety of Talcom?are similar to those of Plavix?for percutaneous coronary interventions.

Objective ATRA can restrain proliferation and promote differentiation in various tumor cells. The aim of the study is to investigate the differentiation characteristics of K562 induced by ATRA. Methods Morphology (Benzidine staining, Wright's staining, NSE staining and NBT recovery test) and flow cytometry were used to observe the differentiation characteristics of K562 after co-incubation with 1 μmol/L and 2.5 μmol/L ATRA for 1 d, 4 ds, and 5 ds. Results Co-incubated for 4 ds, 61.5% K562 cells in the 1 μmol/L ATRA group and 39% K562 cells in the 2.5 μmol/L ATRA group showed some evidence of myeloid maturation, but no evidence of erythroid or monocytoid maturation. Co-incubated for 5 ds, the percentage of differentiated K562 cells was much higher than that in the control group. One day after induction by 1 μmol/L ATRA or 2.5 μmol/L ATRA, the expression of CD13 was 8.0% and 6.7%, respectively, which was higher than that in the control group (2.1%). Five days after induction by 1 μmol/L ATRA or 2.5 μmol/L ATRA, the expression of CD13 increased to 28.1% and 37.8%, respectively, while the expression of CD71 decreased to 1.2% and 0.9% respectively. The differences between the ATRA groups and the control group were significant (P<0.05). CD71 decreased from 9.7% and 10.8% in the 1 μmol/L and 2.5 μmol/L ATRA groups on day 1 to 1.2% and 0.9% on day 5, while the CD13 expression level increased from 8.0% and 6.7% to 28.1% and 37.8%, respectively. Conclusions ATRA can induce K562 to differentiate into myeloid linage.

OBJECTIVETo explore the effect of T(3) on the expression of transferrin receptor (TfR) and ferritin (Fn) in K562 cells and its possible mechanism.

METHODSFlow cytometry was used for the detection of TfR expression, radioimmunoassay for Fn expression, RNA/protein band shift assay for the binding activity of iron regulatory protein (IRP) and iron responsive elements (IRE), and RT-PCR for TfR and Fn mRNA levels.

RESULTSDifferent concentration of T(3) significantly increased Fn expression of K562 cells, especially at 100 nmol/L and 200 nmol/L (p < 0.05). However, T(3) had no effect on TfR expression. T(3) decreased the binding activity between IRP and IRE, particularly at concentration of 50 nmol/L. Different concentration of T(3) increased Fn-H mRNA level at different time point while it had no effect on TfR mRNA level.

CONCLUSIONT(3) increased Fn expression of K562 cells through the possible mechanisms of either the post-transcriptional regulation or transcriptional modulation.

Ferritins ; biosynthesis ; drug effects ; genetics ; Flow Cytometry ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; Radioimmunoassay ; Receptors, Transferrin ; biosynthesis ; drug effects ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Triiodothyronine ; pharmacology

AIMTo investigate whether AcSDKP can inhibit proliferation and collagen synthesis in cultured rat cardiac fibroblasts mediated by PDGF.

METHODSNeonatal rat cardiac fibroblasts were isolated. The cell proliferation was observed by 3H-proline incorporation assay.

RESULTSOn the culture of 0.4% FBS, PDGF stimulated cardiac fibroblasts proliferation and collagen synthesis with a dose-dependent manner at the concentrations from 1 ng/ml to 20 ng/ml, in which 10 ng/ml PDGF reached its peak. AcSDKP at the concentration from 10(-10) mol/L to 10(-8) mol/L could inhibit cardiac fibroblasts proliferation and collagen synthesis mediated by PDGF. 10(-9) mol/L AcSDKP attained its peak on inhibiting cardiac fibroblasts proliferation and collagen synthesis.

CONCLUSIONAcSDKP can inhibit proliferation and collagen synthesis in cultured rat cardiac fibroblasts mediated by PDGF.

Animals ; Cell Proliferation ; Cells, Cultured ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; drug effects ; metabolism ; Myoblasts, Cardiac ; cytology ; drug effects ; metabolism ; Oligopeptides ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Rats, Wistar

BACKGROUNDThe importance of diagnosis of thyroid dysfunction during pregnancy has been widely recognized. Our study was designed to compare two different detection reagents between Abbott and Roche and to establish the gestational related reference intervals for thyroid function tests (TFT) in Chinese women and to assay the reference ranges with the American Thyroid Association recommended standard.

METHODSSerum samples were collected from 693 normal pregnant Chinese women and divided into five groups according to their gestational age: 9-13, 16-20, 24-28, 32-34 and 37-40 weeks. Thyroid stimulating hormone (TSH) and free thyroxine (FT4) levels were determined by two different detection reagents: Abbott Architect I 2000 and Roche Cobas Elecsys 600. The reference ranges of the TFT indexes were calculated according to the National Academy of Clinical Biochemistry (NACB). The 2.5th and 97.5th percentiles of each stage were calculated, and the results were analyzed by one-way analysis of variances, t-test, and Spearman correlation analysis.

RESULTSThyroid hormone levels varied greatly among different gestational stages. TSH levels, as assessed via two different TSH ELISA kits showed consistent changing pattern during pregnancy and displayed linear correlation (P < 0.001). In 9-13 gestational weeks, TSH levels were significantly lower than that of other groups; and in 37-40 gestational weeks, it was higher than that of other groups (all P < 0.001). TSH reference ranges determined by Roche detection reagent in each group were higher than those by Abbott detection reagent (P < 0.01 respectively). FT4 levels were higher in 9-13 gestational weeks than that of other groups (P < 0.001). FT4 levels determined by Roche reagent were higher than Abbott reagent in 9-13 weeks, (P < 0.001), and lower in 24-28 and 37-40 weeks (P < 0.001 and P = 0.016, respectively). The TSH level was correlated with FT4 levels in 9-13 gestational weeks by detection reagents (for Abbott reagent, r=-0.319 for FT4 P < 0.001; for Roche reagent, r=-0.352 for FT4, P <0.001).

CONCLUSIONAccurate evaluation of TFT in pregnant women should be based on the gestational-related reference intervals in Chinese population, and different detection reagents should also establish their own reference intervals.

Adult ; Female ; Gestational Age ; Humans ; Luminescent Measurements ; Pregnancy ; physiology ; Reference Values ; Thyroid Function Tests ; Thyroid Gland ; physiology ; Thyrotropin ; blood ; Thyroxine ; blood

Background/Aims: Renal ischemia followed by reperfusion (I/R) is a leading cause of acute kidney injury (AKI), which is closely associated with high morbidity and mortality. Studies have shown that induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) exert powerful therapeutic effects in renal ischemia. However, the efficacy of iMSC-derived exosomes (iExo) on I/R injuries remains largely unknown. Methods: Human iPSCs were differentiated into iMSCs using a modified one-step method. Ultrafiltration, combined with purification, was used to isolate iExo from iMSCs. iExo was administered following I/R injury in a mouse model. The effect of iExo on I/R injury was assessed through changes in renal function, histology, and expression of oxidative stress, inflammation, and apoptosis markers. Further, we evaluated its association with the extracellular signal-regulated kinase (ERK) 1/2 signaling pathway. Results: Mice subjected to I/R injury exhibited typical AKI patterns; serum creatinine level, tubular necrosis, apoptosis, inflammatory cytokine production, and oxidative stress were markedly increased compared to sham mice. However, treatment with iExo attenuated these changes, significantly improving renal function and tissue damage, similar to the renoprotective effects of iMSCs on I/R injury. Significant induction of activated ERK 1/2 signaling molecules was observed in mice treated with iExo compared to those in the I/R injury group. Conclusions The present study demonstrates that iExo administration ameliorated renal damage following I/R, suggesting that iMSC-derived exosomes may provide a novel therapeutic approach for AKI treatment.