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Objective: In this study, InDel markers were developed based on the high frequency Insertion/Deletion region of chloroplast genome of Ligusticum. Germplasm identification and phylogenetic development of Ligusticum chuanxiong and its common adulterants were studied with universal barcode. Methods: The 26 samples of L. chuanxiong and its common adulterants were amplified and sequenced by eight DNA universal barcodes: ycf1, matK, ITS2, rpoC1, rbcL, rpoB, trnK, and psbA-trnH. Genetic distance statistics, barcoding gap and phylogenetic tree analysis methods were used to study the phylogenetic relationship and phylogeny of L. chuanxiong. At the same time, the evolutionary tree was constructed to study molecular identification of L. chuanxiong and its common adulterants. Results: The results showed that rbcL conserved site was the highest (97.32%) with the highest GC content (44.9%). The rbcL+rpoB fragment had the smallest average intraspecific genetic distance (0.002 5). The psbA-trnH sequence fragment had the largest average interspecific genetic distance (0.429 2). The trnK and rbcL+rpoB sequence had the highest interspecific genetic distance. The overlap of the "barcoding gap" region of psbA-trnH was the least. The species of L. chuanxiong and other adulterated species were not accurately identified by the eight pairs of DNA barcodes. The cluster analysis of 24 InDel markers could accurately identify genuine L. chuanxiong and classify the species of L. chuanxiong and its adulterants into four categories, one of which was genuine L. chuanxiong collected from Sichuan. Conclusion: The ability of InDel markers to identify authentic L. chuanxiong and its common adulterants was higher than that of common barcode. According to the above studies, it is found that it is impossible to distinguish L. chuanxiong and its common adulterants by the traditional DNA barcodes because of the large difference in genetic components. The newly developed InDel molecular markers can effectively identify L. chuanxiong and its commonly used adulterants, and provide an effective method for the genuineness of L. chuanxiong at molecular level.
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Objective · To investigate the effects and mechanisms of pregnancy immune tolerance induced by a novel immunosuppressive agent FTY720 and to provide experimental basis for the clinical treatment of unexplained recurrent spontaneous abortion. Methods · The mice of spontaneous abortion model were used as subjects. The effects of intraperitoneal injection of FTY720 on the embryo loss rate in mice of spontaneous abortion model and on the expression of sphingosine-1-phosphate (S1P) in the decidual tissue were observed. S1P-siRNA lentiviral vectors and S1P-overexpression gene lentiviral vectors were constructed and transfected into dendritic cells (DCs) from mouse bone marrow. The effects of FTY720 on the embryo loss rate in mice of spontaneous abortion model after adoptive transferring of these two types of lentiviral vectors were observed. Results · FTY720 had no significant effect on the embryo loss rate in normal pregnant mice. Intraperitoneal injection of FTY720 significantly reduced the embryo loss rate in mice of spontaneous abortion model. The expression of S1P in the decidual tissue in mice of spontaneous abortion model was low. After adoptive transferring of S1P-siRNA lentiviral vector transfected DCs, FTY720 could slightly reduce the embryo loss rate in mice of abortion mouse model, but the effect was far less than that of before adoptive transferring of S1P-siRNA lentivirus transfected DCs. After adoptive transferring of the S1P-overexpression gene lentiviral vector transfected DCs, FTY720 could significantly reduce the rate of embryo loss in mice of spontaneous abortion model and the effect was more significant than that of before adoptive transfecting of S1P-overexpression lentiviral vector transfected DCs. Conclusion · FTY720 is safe. The induction of pregnancy immune tolerance may be related to the blockage of S1P signaling pathway.
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Objective · To investigate the effects and mechanisms of pregnancy immune tolerance induced by a novel immunosuppressive agent FTY720 and to provide experimental basis for the clinical treatment of unexplained recurrent spontaneous abortion. Methods · The mice of spontaneous abortion model were used as subjects. The effects of intraperitoneal injection of FTY720 on the embryo loss rate in mice of spontaneous abortion model and on the expression of sphingosine-1-phosphate (S1P) in the decidual tissue were observed. S1P-siRNA lentiviral vectors and S1P-overexpression gene lentiviral vectors were constructed and transfected into dendritic cells (DCs) from mouse bone marrow. The effects of FTY720 on the embryo loss rate in mice of spontaneous abortion model after adoptive transferring of these two types of lentiviral vectors were observed. Results · FTY720 had no significant effect on the embryo loss rate in normal pregnant mice. Intraperitoneal injection of FTY720 significantly reduced the embryo loss rate in mice of spontaneous abortion model. The expression of S1P in the decidual tissue in mice of spontaneous abortion model was low. After adoptive transferring of S1P-siRNA lentiviral vector transfected DCs, FTY720 could slightly reduce the embryo loss rate in mice of abortion mouse model, but the effect was far less than that of before adoptive transferring of S1P-siRNA lentivirus transfected DCs. After adoptive transferring of the S1P-overexpression gene lentiviral vector transfected DCs, FTY720 could significantly reduce the rate of embryo loss in mice of spontaneous abortion model and the effect was more significant than that of before adoptive transfecting of S1P-overexpression lentiviral vector transfected DCs. Conclusion · FTY720 is safe. The induction of pregnancy immune tolerance may be related to the blockage of S1P signaling pathway.
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Objective To investigate natural spontaneous menopausal age , menstruation span and their relationship with menarche age and parity in Pudong district of Shanghai. Methods From Jan 2007 to Jul 2008, 15 083 spontaneous menopause women undergoing cervical cancer screening were enrolled in this study. The questionnaire included menarche age, parity, spontaneous menopausal age and menstruation span. Those women were divided into four groups based on age, which were group of 56 -60, 61 -65, 66 -70 and more than 70. Analysis of variance (ANOVA) was used for comparing difference between menopausal age and menstruation span. Multiple factor regressions was used to analyze the relationship between menarche age, parity and menopausal age and menstruation span. Results (1) Spontaneous menopausal age: the minimum was 29 years old, the maximum was 61 years old, and the mean age was (50.6 ±3.7)years old. The mean spontaneous menopause age were (50.9 ± 3.4), ( 50.7 ± 3.7 ), (50.0 ± 4.1 ), (49.6 ±4.0) years in groups of 56 -60, 61 -65, 66 -70 and more than 70 years. With the increasing age range in four groups, the increasing trends of menopausal age were observed, which the difference of 1.36 year was shown between groups of 56 - 60 and more than 70 years. (2) Menstruation span: the mean of menstruation span was (34.3 ± 4.1 ) years, which the minimal age of 12 years and maximal age of 48 years were recorded. (34.6 ± 3.8), (34.3 ± 4.1 ), (33.9 ± 4.6), (33.2 ± 4. 5) were observed in groups of 56 - 60,61 -65, 66 -70 and more than 70 years. With the increasing age range in four groups, the increasing trends of menstruation span were observed, which the difference of 1.41 year was shown between groups of 56 –60 and more than 70 years. (3)The impact of menarche age on menopausal age and menstruation span: there was no correlation between menarche age and menopausal age ( r = 0.02); however, menstruation span was found to be negatively correlated with the menarche age ( r = - 0.43 ). (4) The impact of parity on menopausal age and menstruation span: the mean menopausal age of women who had 1 -2 deliveries was significantly higher than those had no delivery or more than 3 deliveries ( P < 0.05 ). However, there was no difference in menopausal age between women with 1 and 2 deliveries or between women without delivery and more than 3 deliveries (P > 0.05). Menstruation span of women with 1 delivery was significantly longer that those with more than 1 delivery( P < 0.05 ), similarly, women with 2 deliveries had longer menstruation span than women without delivery or more than 3 deliveries(P < 0.05 ). There were no difference in menstruation span between women with more than 3 deliveries and without delivery ( P >0.05 ). (5) Multifactor regression analysis for menstruation span: menarche age was correlated with menstruation span negatively ( r = - 0.97,P <0.001 ). There was significantly different menstruation span between group of 61 -65, 66 -70 or more than 70 years and group of 56-60 (r= -0. 18, P=0.020; r= -0.78,P <0.001 and r= - 1.23,P<0.001). Menstruation span in women with 1 -2 deliveries was significantly longer than that of women without delivery or more than 3 deliveries. (6)Multifactor logistic analysis of menopausal age: there was no association between menarche age and menopausal age, however, significant differences were found in mean menopausal age between different groups, which show that menopausal age of group 56 - 60 years was significant higher than the other groups, including age-group 61 -65 years ,66 -70 years and over 70 years ( r = - 0.18, P = 0.020; r = - 0.78,P < 0.001; r = - 1.23, P < 0.001 ). Menopausal age in women with 1 - 2 deliveries was significantly higher than those of women without delivery or with more than 3 deliveries,however, no difference between women with 1 and 2 deliveries or between women without deliveries and more than 3 deliveries was observed. Conclusion (1) Menopausal age and menstruation span exhibited increasing trends in Pudong district of Shanghai. (2) Menarche age and parity were the important factors influencing menopausal age and menstruation span. (3) With younger age of menarche, the menstruation span become longer. (4) Deliveries of 1 -2 times can significantly delay the menopause and prolong menstruation span, however, the multiple deliveries ( ≥ 3 times) had no significant impact on menopausal age and menstruation span.
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Objective To evaluate the clinical value of ultrasonography detecting the poster urethrovesical angle(PUVA) in diagnosis of stress urinary incontinence(SUI).Methods From Jan.2006 to Dec.2008,the PUVA in resting phase (PUVA-r) and stress phase( PUVA-s) between with SUI and 100 healthy women were measured by color Doppler ultrasound,including 57 cases in SUI degree Ⅰ,22 cases in SUI degree Ⅱ,5 cases in SUI degree Ⅲ.Results (1) The PUVA-r and PUVA-s were (130±29) and (158±36)in SUI patients,which were significantly higher (113±19)and (115±23)in control group (P<0.01).(2) Correlation analysis showed that PUVA-s was positively correlated with the severity of SUI ( P < 0.01).(3) The area under the receiver operating characteristic curves (ROC) of PUVA-s was 0.82 (95% CI:0.76-0.89) between SUI patients and healthy women.When PUVA-s ≥ 140 ° was chosed as the cut-off value,the specificity,accuracy rate and positive predictive value in diagnosis of SUI were 84%,79%,81%,which were significantly higher than those when cut-off value of PUVA-s ≥ 120(54%,68%,62%; P < 0.05 ).Conclusion The PUVA-s in SUI patients are significantly increased and is positively correlated with the severity of SUI,which are indicated that PUVA-s ≥ 140should be used as cut-off value in diagnosis of SUI by ultrasonography.
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Objective To survey age of menarehe in Pudong district in Shanghai. Methods Data in this study were derived from 56 924 women at age of 20 -81 years in screening for cervical cancer between January 2007 and July 2008 in Pudong district. The age of menarche were recorded in a questionnaire. To investigate the trends in age at menarehe in different socioeconomic status, the subjects were divided into 12 groups in 5-year birth cohorts. The mean menarche age in each group was analyzed by analysis of variance(ANOVA). The percentage of menarche age at 10- 12 years and more than 18 years was analyzed by χ2 method. Results (1 ) The minimum age of menarcbe recorded is 10 years old, and the maximum is 28 years old, with average age of menarche at 15.7 years. In all groups, the smallest average age of menarcbe is 14. 6 years in 26 - 30 years old age group, while the biggest average age of menarche age is 16. 5 years in ＞ 75 years old group; The difference showed statistical significance (P ＜ 0. 01 ). (2) The percentages for participants with early menarehe age (10 - 12 years old) or late menarehe age (＞ 18-year-old menarche) were 1.82% (1034/56 924 ) and 5.20 % (2959/56 924 ) respectively. However, the maximum percentage for early menarche was recorded in 31 -35 years old group (4. 45% ,197/4431 ), only 0. 84% (10/1191 ) of participants in ＞75 years old group was classified as early menarebe. Meanwhile, the lowest percentage for late menarehe was 0. 38% (17/4431 ) in 31 - 35 years old group, and the highest percentage was 14. 70% (91/619) in ＞ 75 years old group. The changes in the percentages for early menarche or late menarche are significantly associated with age differences (P ＜ 0. 01 ). Conclusion The study suggested that the average of onset age of menarche in Pudong district has declined over the past decades in an age-based way, accompanied with the increase of the percentage for early menarche and the decrease of percentage for late menarehe.
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<p><b>BACKGROUND</b>Chemokines and their receptors have been a research focus in transplantation immunology. Chemokines and their receptors play a role in lymphocyte recruitment and differentiation process. This study aimed to observe whether IL-4 and IL-10 may regulate the expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4(+) T cells in CBA/JxDBA/2 mouse model and to explore the role of CCR3, CCR5, CXCR3 in immune tolerance in pregnancy.</p><p><b>METHODS</b>The mouse model of spontaneous abortion (CBA/JxDBA/2) and the normal pregnant mouse model (CBA/JxBALB/c) were used. CBA/JxDBA/2 mice were injected with IL-4 (CBA/JxDBA/2-IL-4), IL-4 and IL-10 (CBA/JxDBA/2-IL-4+IL-10), or normal saline (CBA/JxDBA/2-NS) as a control. The expression of CCR3, CCR5 and CXCR3 on CD4(+) T cells from mouse peripheral blood was measured by the double-labelled FCM method, and the embryo resorption rate was also examined.</p><p><b>RESULTS</b>The embryo resorption rate in the CBA/JxDBA/2 group without any treatment was significantly higher than that in the CBA/JxBALB/c group (17.9% vs 3.7%, P < 0.01). The embryo resorption rate in the CBA/JxDBA/2 group immunized with IL-4 or IL-4 together with IL-10 was significantly decreased, compared with that in the control and NS groups respectively. CCR3 expression on CD4(+) T cells in the CBA/JxDBA/2 group without any treatment was significantly lower than that in the CBA/JxBALB/c group (0.3738 +/- 0.3575 vs 1.2190 +/- 0.2772, P < 0.01); both CCR5 (3.0900 +/- 1.5603 vs 1.2390 +/- 0.6361, P < 0.01) and CXCR3 (2.4715 +/- 0.9074 vs 0.9200 +/- 0.5585, P < 0.01) expressions on CD4(+) T cells of the CBA/JxDBA/2 group without any treatment were significantly higher than those of the CBA/JxBALB/c group. Significant up-regulation of CCR3 and down-regulation of CXCR3 were found in the CBA/JxDBA/2 group treated with IL-4 (CCR3: 2.0360 +/- 0.6944, CXCR3: 1.3510 +/- 0.5263, P < 0.01) or IL-4 and IL-10 (CCR3: 1.8160 +/- 1.0947, CXCR3:1.0940 +/- 0.7168, P < 0.01). Because of the CCR5, IL-4 and IL-10 (1.9400 +/- 0.8504 vs 3.0900 +/- 1.5603, P < 0.05), but IL-4 alone (2.5310 +/- 1.3595 vs 3.0900 +/- 1.5603, P > 0.05) treatment significantly decreased the expression of CCR5 in CBA/JxDBA/2.</p><p><b>CONCLUSIONS</b>The abnormal expression of CCR3, CCR5 and CXCR3 on CD4(+) T cells may play an important role in the pathogenesis of spontaneous abortion. The pregnancy immune tolerance may be induced through selective induction of CCR3, CCR5 and CXCR3 expressions by IL-4 together with IL-10.</p>
Subject(s)
Animals , Female , Mice , Pregnancy , CD4-Positive T-Lymphocytes , Metabolism , Cells, Cultured , Embryo Loss , Metabolism , Embryo, Mammalian , Interleukin-10 , Pharmacology , Interleukin-4 , Pharmacology , Mice, Inbred BALB C , Receptors, CCR3 , Metabolism , Receptors, CCR5 , Metabolism , Receptors, CXCR3 , MetabolismABSTRACT
Objective To investigate the role of vascular endothelial growth factor (VEGF)in the pathogenesis of bronchial asthma and chronic obstructive pulmonary disease (COPD). Methods Thirty patients with stable bronchial asthma (asthma group), 28 patients with stable COPD (COPD group), and 24 healthy subjects (control group) were studied. Lung function, inflammatory cell differentials in sputum and the level of VEGF in induced sputum were determined by induced sputum method and ELISA method respectively. Results The levels of eosinophils in induced sputum in asthma group were 0.9 (0.4-1.4) × 109/L, significantly higher than those in COPD greup[0.1 (0-0.2)×109/L], and control group[0.0(0-0.1) × 109/L] (P < 0.05). The levels of neutrophils in COPD group were 2.3 (1.8-2.8) × 109/L, significantly higher than those in asthma group [1.1 (0.2-1.9) × 109/L], and control group [1.0(0.8-1.2) × 109/L] (P < 0.05). The levels of VEGF in asthma group, COPD group and control group were (2.3 ± 0.5), (0.3 ± 0.1), (0.9 ± 0.2) μg/L, respectively, and significant difference was showed between each group (P < 0.05). The VEGF level in induced sputum was positively correlated with the number of eosinophils in induced sputum in asthma group (γ = 0.62,P < 0.05), and was negatively correlated with FEV<,1>% (γ =-0.56, P < 0.05). The VEGF level in induced sputum was positively correlated with FEV1% in COPD group (γ = 0.43, P < 0.05), and was not correlated with the number of neutrophils in induced sputum (γ = 0.21, P > 0.05). Conclusions The concentration of VEGF in induced sputum increases in patients with bronchial asthma. VEGF may take part in the airway inflammatory development of asthma. The concentration of VEGF in induced sputum decreases in patients with COPD. VEGF may take part in the incidence of COPD.
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Objective To study the mechanism of pregnancy immune tolerance induced by adoptive transferring of FasL gene-modified dendritic cells(DC). Methods The mouse models of spontaneous abortion(CBA/J ? DBA/2) and normal pregnancy(CBA/J ? BALB/c) were established.Five different experimental groups were included: mice of normal pregnancy(CBA/J?BALB/c)(n=17),served as control group;mice of spontaneous abortion without treatment(CBA/J?DBA/2)(n=37),mice injected with DC culture medium(DCCM)(n=25);mice immunized with empty plasmid pcDNA3.1-DC(n=6);and mice immunized with pcDNA3.1-FasL-DC(n=5).Embryo resorption rates of pregnant mice in each groups were observed.Annexin V-FITC was used to detect the apoptosis of T cells.Immunohistochemical staining(SABC) was used to detect the expression of FasL in decidual membranes of pregnant mice. ResultsThe embryo resorption rate of mice immunized with FasL-DC was decreased significantly as compared with that of mice of spontaneous abortion without treatment,DCCM group and immunized with pcDNA3.1-DC(P0.05). Conclusion The decreased apoptosis rate of peripheral T cells and the reduced expression of FasL in decidual membranes may be an important mechanism for the pathogenesis of abortion.Adoptive transfer of FasL gene-modified DC may induce pregnancy immune tolerance by increasing FasL expression of maternal-fetal interface and decreasing embryo resorption rate.
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<p><b>OBJECTIVES</b>To confirm previous whole-genome scan results of mapping type 2 diabetes susceptibility genes in chromosome 1 in Northern Chinese Han population by conducting a new genome scan with both an enlarged number of type 2 diabetes families and a new set of microsatellite markers.</p><p><b>METHODS</b>A genome scan method was applied. After multiplexed PCR, electrophoreses, genescan and genotyping analysis, size informations for all loci were obtained, and a further study was done using both parametric and non-parametric linkage analysis to calculate the P-values and Z-values of these loci.</p><p><b>RESULTS</b>A total of 34 microsatellite markers distributed within 5 regions along chromosome 1 were surveyed, and 12,000 genotypes were screened. Evidence of linkage with diabetes was identified for 8 of the 34 loci (all the P-values of the 8 loci distributed in 3 regions were lower than 0.05, and the highest Z-value was 2.17). Interestingly, all the 5 markers at the P terminal 1p36.3-1p36.23 region, spanning a long range of 16.9 cM, suggested to be linked with the disease. The results of the other two regions were not consistent with the previous ones.</p><p><b>CONCLUSIONS</b>The study results have confirmed those gained in the previous genome-wide scan. The fact that all 5 loci at the P terminal region displayed linkage with diabetes suggests that more than 1 susceptibility gene may reside in this region.</p>