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1.
In this study, we identified a sex-specific DNA marker using randomly amplified polymorphic DNA (RAPD) fingerprinting in Yellow River carps (Cyprinus carpio from the Yellow River). Two hundred and twenty random primers were used in pooled DNA samples and individual DNA samples from male and female fish for RAPD fingerprinting. When using the primer S2107, a novel sex-specific PCR product was identified in all male individuals. DNA sequencing revealed that this 909 bp long DNA fragment has a low similarity to a repetitive sequence in zebrafish Danio rerio. In order to confirm the amplification results, two primers were designed within the male-specific sequence in order to amplify the sex-specific fragment from genomic DNA of male and female carps for sexing by PCR. The results indicated bands specific for males but not females. Respective results were obtained in dot blot and Southern blot hybridization experiments when using this sex-specific fragment as the probe. The sex-specific pattern was observed in 30 individuals from three separate common carp stocks, suggesting that the sequence is conserved in common carp species. However, no hybridization signals were found in grass carps Ctenopharyngodon idella, which are related to common carps. We submit to use this sex-specific fragment as a marker to rapidly and accurately identify the gender of Yellow River C. carpio. Furthermore, the sex-specific chromosome region may be characterized and used to study mechanisms of chromosome evolution in this fish species.  相似文献   

2.
Due to over increasing demand, huge collection and habitat destruction wild gingers having future medicinal potential are on way to extinction. Conservation of germplasm in clonal repositories by conventional methods is very difficult due to greater susceptibility to rhizome-rot disease. In vitro conservation strategy has so far been made for a few species. Thus the present study aiming at in vitro conservation of nine species and subsequent stability assessment of plantlets using efficient molecular markers assumes significance. Murashige and Skoog media with different growth regulators were standardized for conservation. Conservation period without sub culturing for all species was in a range of 8–12 months. Survival was more than 80 % up to three years. On the basis of 25 random amplified polymorphic DNA and ten Inter simple sequence repeat primers no significant variation was observed among the in vitro conserved and control plants. The micropropagated plants are morphologically and genetically uniform when compared to their field grown mother plants. Thus, the report has immense potential towards the conservation of genetic resources for future breeding and improvement programme by continuous supply of genetically uniform plantlets.  相似文献   

3.
An efficient, reliable and reproducible plant regeneration protocol was developed for Lawsonia inermis L. using mature nodal explants. Shoot proliferation (81.6 %) with 7.8 shoots/explant was achieved on Murashige and Skoog’s (MS) medium supplemented with 1.0 mg l?1 6-benzyladenine. Shoot numbers were up-scaled by inducing multiple shoots from axenic nodal segments derived from the primary shoots on the shoot regeneration medium. Thus the authors could achieve ca. 129–134 shoots from single nodal explant. Ninety-three percent rooting of in vitro regenerated shoots was achieved on growth regulator free half-strength MS medium. Regenerated plantlets were successfully transferred to soil with 85 % survival rate. Genetic stability analyses of the in vitro regenerated plants using random amplified polymorphic DNA and inter simple sequence repeat markers revealed a homogeneous amplification profile for all micropropagated plants. This is the first report that evaluates the use of molecular markers to establish genetic fidelity of micropropagated L. inermis for the rapid clonal multiplication and true-to-type production of plant for attaining the ever increasing demand in pharmaceutical industries.  相似文献   

4.
Left ventricular (LV) twist is emerging as a marker of global LV contractility after acute myocardial infarction (AMI). This study aimed to describe stress-induced changes in LV twist during dobutamine stress echocardiography (DSE) after AMI and investigate their association with LV reverse remodeling at 6 months follow-up. In 82 consecutive first AMI patients (61 ± 12 years, 85 % male) treated with primary percutaneous coronary intervention, DSE was performed at 3 months follow-up. Two-dimensional speckle-tracking-derived apical and basal rotation and LV twist were calculated at rest, low- and peak-dose stages. LV reverse remodeling was defined as ≥10 % decrease in LV end-systolic volume between baseline and 6 months follow-up. Patterns of LV twist response on DSE consisted of either a progressive increase throughout each stage (n = 18), an increase at either low- or peak-dose (n = 53) or no significant increase (n = 11). LV reverse remodeling occurred in 28 (34 %) patients, who showed significantly higher peak-dose LV twist (8.51° vs. 6.69°, p = 0.03) and more frequently progressive LV twist increase from rest to peak-dose (39 vs. 13 %, p < 0.01) compared to patients without reverse remodeling. Furthermore, increase in LV twist from rest to peak-dose was the only independent predictor of LV reverse remodeling at 6 months follow-up (OR 1.3, 95 % CI 1.1?1.5, p = 0.005). Both the pattern of progressive increase in LV twist and the stress-induced increment in LV twist on DSE are significantly associated with LV reverse remodeling at 6 month follow-up after AMI, suggesting its potential use as a novel marker of contractile reserve.  相似文献   

5.
DNA methylation has been implicated in the regulation of gene expression, genome imprinting, and chromatin remodeling in eukaryotes. In this study, we analyzed possible alterations in levels and patterns of cytosine methylation in male and female spinach plants after treatment with demethylation agent 5-azacytidine (5-azaC) using two methods: (1) direct determination of 5-methylcytidine (5 mC) amounts in genomic DNA by high-performance liquid chromatography (HPLC) separation and quantification of nucleosides and (2) methylation-sensitive inter-simple sequence repeat (MS-ISSR) technique. HPLC analysis revealed that the DNA methylation events in male and female spinach leaves markedly decreased upon 30 μM 5-azaC treatment, and the methylation level gradually decreased with the increase in 5-azaC concentration. To study the altered DNA methylation patterns in spinach after 5-azaC treatment, untreated and 500 μM 5-azaC-treated samples were analyzed by MS-ISSR assay. A total of 385 informative profiles were resolved using 35 ISSR primer sets. MS-ISSR analysis showed various altered methylation patterns between untreated and 5-azaC-treated spinach plants. These alterations were mainly demethylation events, which were largely consistent with the HPLC results. Both HPLC and MS-ISSR analyses showed that the changes in DNA methylation levels and patterns were similar in male and female spinach leaves, which implies that sex was not the main factor influencing DNA methylation levels and patterns in the vegetative organs of spinach. This study could provide a molecular basis of the altered DNA methylation induced by 5-azaC, and lay a foundation for further investigation of the relationship between methylation and sex determination and development in this dioecious plant spinach.  相似文献   

6.
In search for safe and eco-friendly management of rodent pests a number of phyto-chemicals have been evaluated as repellent, antifertility agent, antifeedant and toxicant etc. Protein rich residue of seed of Jojoba (Simmondisa chinensis) known as defatted jojoba meal is reported to possess marked suppressive effect on food intake in a variety of animal species. In present study the defatted jojoba meal at a concentration of 5, 10, 15, 20, 25 and 30 % in pearl millet bait (w/w basis) was evaluated for its anti-rodent properties against Tatera indica, a predominant rodent pest of Indian subcontinent. Under no-choice condition the bait consumption during treatment period of 7 days was significantly reduced (3.01 ± 0.40–4.76 ± 0.14 g/100 g bwt/day) in comparison to pre-treatment plain bait consumption (9.33 ± 0.73–10.37 ± 0.29 g/100 g bwt/day). At higher dosages 50 % gerbils died. Besides, weight reduction in the range of 10.63–16.67 % was also observed. In choice test mean consumption of treated food was also significantly reduced (0.64 ± 0.28–1.44 ± 0.20 g/100 g bwt/day) in comparison to plain bait (5.80 ± 0.18–8.80 ± 0.29 g/100 g bwt/day) during 7 days of treatment period. Jojoba treated pearl millet bait when offered with plain sorghum bait, after an exposure period of 5 days left an imprinting effect on Indian gerbil which led to aversion towards plain pearl millet bait for 10–11 days. The findings showed that jojoba defatted meal exert strong dose dependant deterrence in Indian gerbils and also induces condition aversion learning behavior in the gerbils.  相似文献   

7.
广东汉族人群St14(DXS52)VNTR与血友病A携带者的检测   总被引:2,自引:0,他引:2  
目的 研究广东正常人群St14(DXS52)位点的遗传多态性,为血友病A的基因诊断提供依据。方法 采用扩增片段长度多态性(Amp-FLP)分析技术,检测了广东地区无血缘关系的正常汉族个体125名,男21名,女104名,总计229条X染色体。利用此数目可变串联重复序列(VNTR)多态作为遗传标记,对4个血友病A家系进行连锁分析。结果 共栓出11种等位片段,其片段大小为700 ̄1810bp,基因频率分  相似文献   

8.
A chitinolytic Streptomyces strain isolated from an alkaline habitat produced six different isozymes of chitinases. PCR amplification of DNA with chitinase domain specific primers yielded six amplicons of which, a 0.8 Kb fragment was cloned in pQE-30UA direct cloning vector and transformed into E. coli M15 cells (pREP4). The recombinant homodimer protein had a molecular mass of 44 kDa, and the 22 kDa monomers displayed 45 and 60 % activity in the presence of reducing agents. The size of the monomers is close to the predicted putative ORF of 17.8 kDa. The enzyme exhibits extreme pH and temperature optima of 10.0 and 70 °C respectively making it favorable for industrial applications. Its gene sequence revealed no homology with the reported N-acetylglucosaminidases suggesting that it could have novel attributes. This enzyme could be useful in the large scale production of N-acetylglucosamine which is currently having numerous therapeutic and commercial applications.  相似文献   

9.
Increased pressure pain sensitivity and impaired descending pain control have been associated with chronic pain, but knowledge on the variability in the adult general population is lacking. Pressure pain thresholds (PPTs) and descending pain control assessed using conditioned pain modulation (CPM) were recorded in a randomly selected sample (n = 2,199, 53% female) of the Danish adult general population aged 18 to 70 years. PPTs were recorded over the tibialis anterior muscle and the upper trapezius muscle. CPM was defined as the difference between PPT assessments before and during conditioning with cold pressor pain (hand) for 2 minutes. Conditioning pain intensity was assessed using a visual analog scale and questionnaire data were collected. Female sex (P < .001) and younger age (P ≤ .02) was associated with lower PPTs at both body sites. For the trapezius muscle, high perceived stress was associated with lower PPTs (P < .02), whereas an interaction was found between body mass index and sex. CPM potency was lower in female compared with male participants (P ≤ .003), whereas no association with age was found. Higher level of education (P ≤ .05), premature withdrawal from the cold pressor test (P ≤ .02), and high visual analog scale score (P ≤ .02) were associated with a larger CPM response.

Perspective

Data from this large population-based study provide new insight into the gender and age variation in pain sensitivity and CPM response. Decreased CPM potency and increased pain sensitivity in female participants were found, emphasizing the need to improve the understanding of its clinical consequences.  相似文献   

10.
An unusual lineage of alpha/beta T cells that contains autoreactive cells.   总被引:5,自引:13,他引:5  
In male mice that express a transgenic alpha/beta T cell receptor (TCR) specific for a male-specific peptide presented by class I Db major histocompatibility complex (MHC) molecules, we describe an unusual lineage of alpha/beta T cells that are thymus dependent but do not require selection by Db MHC molecules on thymic epithelium in the absence of the specific peptide (positive selection). These cells express the transgenic alpha/beta TCR and have the CD4-8- or CD4-8low phenotype. Cells with the latter phenotype are only detected when hemopoietic cells express both the male-specific peptide as well as Db MHC molecules. In fact, these cells are autoreactive, as they expand relatively slowly after transfer into male nude mice. Also in male but not female alpha/beta TCR transgenic mice, the CD8+ cells with the transgenic TCR bear the Pgp1 marker characteristic of mature T cells activated by antigen. CD4-8- as well as CD4-8low cells do not respond significantly when cultured with male stimulator cells but proliferate vigorously when stimulated by TCR antibodies. By this latter criterion, cells in the periphery of male alpha/beta TCR transgenic mice differ from mature male-specific T cells from female alpha/beta TCR transgenic, which become intrinsically anergic when transferred into male nude mice and cannot be stimulated significantly by TCR antibodies. Thus, intrathymic deletion does not eliminate all autoreactive T cells and it is possible that cells with an apparently "benign" autoreactivity may be involved in certain forms of autoimmunity.  相似文献   

11.
Navigation of intracardiac catheters by echocardiography is challenging because of the fundamental limitations of B-mode ultrasonography. We describe a catheter fitted with a piezoelectric crystal, which vibrates and produces an instantaneous marker in color flow Doppler scans. The navigation learning curve was explored first in six pigs. Accuracy and precision of targeting with the navigation marker “off” (i.e., B-mode imaging) and “on” were assessed in another six pigs. Paired comparisons confirmed significantly (p = 0.04) shorter mean distances achieved in each pig with the color Doppler marker. Pooled (mean ± standard deviation) distance of the catheter tip from the target crystal was 5.27 ± 1.62 mm by B-mode guidance and 3.66 ± 1.45 mm by color Doppler marker navigation. Dye injection targeted into the ischemic border zone was successful in 8 of 10 pigs. Intracardiac catheter navigation with color Doppler ultrasonography is more accurate compared with conventional guidance by B-mode imaging.  相似文献   

12.

Objective

To compare characteristics and outcome of patients with re-stenoses after prior carotid artery stenting (CAS) treated with repeat carotid interventions (Re-CI) with CAS for de novo lesions.

Background

The treatment of re-stenosis is a major problem in vascular interventions. Patients with re-stenoses after prior CAS treated with Re-CI are not well defined.

Methods

We analyzed data from the prospective ALKK CAS Registry.

Results

Out of 3,817 CAS procedures 95 were intended in 93 patients (2.5%) for a restenosis after prior CAS and 3,722 CAS in 3,655 patients (97.5%) for a de novo stenosis. There was no difference in age (p = 0.302) or distribution of gender (p = 0.545) between the two groups. Patients treated for a restenosis after CAS were less likely to be treated for a symptomatic lesion (22.7 vs. 40.1%, p = 0.001). Coronary heart disease (p = 0.017), peripheral arterial disease (p < 0.001) as well as diabetes mellitus (p = 0.004) were more prevalent in the restenosis group. Lesions were less complicated in restenosis patients, with less ulcers (7.4 vs. 19.9%, p = 0.003) and less severe calcifications (7.4 vs. 23.6%, p < 0.001). The intended interventions were more often not performed in the Re-CI group (9.5 vs. 3.3%; p = 0.001). In-hospital, the stroke or death rate was 0% in the Re-CI group as compared to 3.1% in the de novo group (p = 0.115).

Conclusions

Patients treated with Re-CI for repeat stenoses after prior CAS represent 2.5% of current CAS patients. Although representing a subgroup with more concomitant diseases, Re-CI seems to be associated with lower event rates as compared to CAS for de novo lesions.  相似文献   

13.
Bordetella pertussis was detected by spectrofluorometry following PCR incorporating a molecular beacon probe in the reaction. A DNA fragment from the tandem repeat sequence region (IS 481) of the genome of B. pertussis was amplified in presence of the probe complementary to an internal segment of the amplified DNA fragment. Fluorescein (FAM) and DABCYL were used as the fluorophore and quencher in the probe. The probe was characterized for its signal to noise ratio by homogeneous solution hybridization with a complementary oligonucleotide. Measurement of fluorescent signal at the emission maxima of FAM, immediately after a PCR was used to detect the B. pertussis target, with no additional steps. Presence of B. pertussis in a sample was also examined by agarose gel electrophoresis of the PCR product. A serial diluted stock of B. pertussis (ATCC strain #9797) and fourteen clinical isolates of B. pertussis were examined. The sensitivity of detection by fluorescent measurement was found to be at least in the range of 0.01-0.1 CFU per 10 microl of the sample and was equal to or better than that detected by agarose gel analysis.  相似文献   

14.
Antheraea frithi Moore, the source of wild tasar silk, is an indigenous wild silk moth of north-east India. The detailed life history, bionomics and rearing performance of A. frithi in relation to a newly reported food plant, Terminalia arjuna (Roxb.) has been described in the present study. Life cycle of A. frithi has five larval instars. The first instar larva measures 1.98 ± 0.13 cm × 0.198 ± 0.010 cm in length and breadth and 0.0142 ± 0.002 g in weight while the fifth instar larva measures 6.98 ± 0.18 cm × 1.12 ± 0.13 cm in length and breadth and 4.76 ± 0.45 g in weight. Wing span of male and female is recorded as 11.4–13.5 and 13.0–14.7 cm respectively. The incubation period (6–7 days), larval duration (35–39 days), fecundity (162–176 eggs), hatching (78–81 %), effective rate of rearing (57–59 %) and cocoon yield (67–79 nos) has been recorded. The reelable cocoons with a filament length (mts) and filament denier (d) is 493–575 m and 4.6–5.3 respectively. The shell weight (g) of male and female cocoons measures as 0.19–0.39 and 0.2–0.6, respectively. The shell ratio (%) for male and female cocoons stands as 8.1–8.4 and 9.4–9.8 % respectively.  相似文献   

15.
Identifying Lactobacillus species using only phenotypic and genotypic (16S rDNA sequence analysis) techniques yields inaccurate results. The objective of this study was to develop species-specific primers based on randomly amplified polymorphic DNA (RAPD) fingerprinting to distinguish species within the closely related Lactobacillus plantarum group. One of these primers, OPD-3, produced a species-specific band that was found only in the tested Lactobacillus pentosus. This specific fragment was isolated from agarose gel and ligated into a vector for DNA sequencing. A pair of primers, SpOPD3Lpen-F1/R1, that were highly specific sequence-characterized-amplified-regions (SCARs) were designed according to the nucleotide sequences of the specific RAPD marker. These primers were used for PCR analysis of the template DNA of the Lactobacillus strains, and a single 542 bp species-specific band was found only in L. pentosus. Using PCR, a novel species-specific primer pair is shown to rapidly, accurately and effectively distinguish L. pentosus from other species in the L. plantarum group of probiotic bacteria.  相似文献   

16.
To evaluate left and right ventricular myocardial performance using pulsed-tissue Doppler imaging (TDI) and its relation to BNP levels in patients with beta-thalassaemia major (ß-TM). We enrolled 36 thalassaemic patients (21 male, 15 female; mean age: 14.2 ± 4.1 years) with normal left ventricular systolic and diastolic functions with conventional echocardiography and 30 healthy control subjects (18 male, 12 female, and 12.5 ± 4.2 years). Myocardial performance indexes (MPI) of left ventricular (LV) lateral wall, interventricular septum (IVS) and right ventricular (RV) lateral wall were calculated with TDI. Plasma BNP levels were measured in all patients. MPIs and other echocardiographic parameters of patients with ß-TM were compared with control group. All the patients’ plasma BNP levels were within normal limits. There were no differences between conventional echocardiographic parameters of patients and control group. MPI of LV, IVS, and RV of patients were significantly higher than control group (= 0.01, and < 0.01, and < 0.001, respectively). Our study confirms that MPI obtained by TDI seems to be an early sensitive parameter of cardiac dysfunction in ß-TM. We concluded that MPI obtained by TDI may be an adjunctive parameter to conventional echocardiography for detecting early myocardial damage.  相似文献   

17.
Microvolt T-wave alternans (MTWA) is an electrocardiographic marker for predicting sudden cardiac death. In this study, we aimed to study the relation between MTWA and scar assessed with cardiac magnetic resonance imaging (CMR) in patients with ischemic cardiomyopathy (ICM) or dilated cardiomyopathy (DCM). Sixty-eight patients with positive or negative MTWA and analysable CMR examination were included. Using CMR and the delayed enhancement technique, left ventricular ejection fraction (LVEF), volumes, wall motion and scar characteristics were assessed. Overall, positive MTWA (n = 40) was related to male gender (p = 0.04), lower LVEF (p = 0.04) and increased left ventricular end-diastolic volume (LVEDV) (p < 0.01). After multivariate analysis, male gender (p = 0.01) and lower LVEF remained significant (p = 0.02). Scar characteristics (presence, transmurality, and scar score) were not related to MTWA (all p > 0.5). In the patients with ICM (n = 40) scar was detected in 38. Positive MTWA (n = 18) was related to higher LVEDV (p = 0.05). In patients with DCM (n = 28), scar was detected in 11. Trends were found between positive MTWA (n = 15) and male gender (p = 0.10), lower LVEF (p = 0.10), and higher LVEDV (p = 0.09). In both subgroups, the presence, transmurality or extent of scar was not related to MTWA (all p > 0.45). In this small study, neither in patients with ICM or DCM a relation was found between the occurrence of MTWA and the presence, transmurality or extent of myocardial scar. Overall there was a significant relation between heart failure remodeling parameters and positive MTWA.  相似文献   

18.
To evaluate long-term changes in diffuse myocardial fibrosis using cardiac magnetic resonance (CMR) with late gadolinium enhancement (LGE) and T1 mapping. Patients with chronic stable cardiomyopathy and stable clinical status (n = 52) underwent repeat CMR at a 6 month or greater follow up interval and had LGE and left ventricular (LV) T1 mapping CMR. Diffuse myocardial fibrosis (excluding areas of focal myocardial scar) was assessed by post gadolinium myocardial T1 times. Mean baseline age of 52 patients (66 % male) was 35 ± 19 years with a mean interval between CMR examinations of 2.0 ± 0.8 years. CMR parameters, including LV mass and ejection fraction, showed no change at follow-up CMR (p > 0.05). LVT1 times (excluding focal scar) decreased over the study interval (from 468 ± 106 to 434 ± 82 ms, p = 0.049). 38 Patients had no visual LGE?, while 14 were LGE+. For LGE? patients, greater change in LV mass and end systolic volume index were associated with change in T1 time (β = ?2.03 ms/g/m2, p = 0.035 and β = 2.1 ms/mL/m2, p = 0.029, respectively). For LGE+ patients, scar size was stable between CMR1 and CMR2 (10.7 ± 13.8 and 11.5 ± 13.9 g, respectively, p = 0.32). These results suggest that diffuse myocardial fibrosis, as assessed by T1 mapping, progresses over time in patients with chronic stable cardiomyopathy.  相似文献   

19.
Gender- and age-related changes of left ventricular (LV) function and dimensions have not been elucidated in a large population by gated SPECT. Thus, the aim of this study was to derive male and female reference limits for left ventricular functional parameters, and determine the effect of age on LV dimensions and systolic function for this imaging modality. 1,639 (53 % males) subjects without cardiovascular disease who underwent cardiac SPECT between January 2002 and June 2012 were included in this study. Mean age at presentation was 61 ± 12 years (range 18–92 years). A significant effect of age (p = 0.011) and gender (p < 0.0001) on resting LV ejection fraction (LVEF) was observed, with an increase in LVEF with age being more pronounced in women (ΔB-coefficient: ?0.088, p = 0.011). Overall, mean LVEF was higher in women compared to men (70.3 ± 8.6 % vs. 64.4 ± 7.5 %, p < 0.0001). LVEF after pharmacological stress with adenosine was significantly lower than at rest in both women and men (?LVEF = 1.1 % in males and ?LVEF = 1.6 % in females, p = 0.01), which was the result of a significant increase in end-systolic volume after stress (p = 0.0001). With advancing age an increase in LVEF was observed that was more pronounced in women than in men. These findings indicate that the evaluation of cardiac function and volumes of patients by gated SPECT should consider age- and gender-matched normative values.  相似文献   

20.

Purpose

To establish whether in critically ill patients without sepsis at intensive care unit (ICU) admission the percentage immature platelet fraction (IPF%) is a cellular marker predicting sepsis to verify a possible correlation between IPF% changes and manifest sepsis and describe the IPF% time course after ICU admission.

Methods

Prospective, observational 7-day study of 64 adult patients admitted to a general ICU at a University Hospital with no sepsis criteria. We measured daily IPF%, procalcitonin (PCT), C-reactive protein, platelets, white blood cell count and coagulation variables. Thirty-one patients with sepsis at ICU admission were studied as controls.

Results

The only variable we tested at ICU admission that predicted sepsis was plasma IPF% (p < 0.001; >4.7 %: sensitivity 56.2 % IC 37.7–73.6; specificity 90.0 % IC 73.4–97.8). IPF% and PCT values were higher for the patients who had sepsis at admission and during the study than in patients in whom sepsis never developed (IPF%: p = 0.017; PCT: p = 0.030). Among the outcome variables, logistic regression was identified as the only variable related to the development of sepsis, IPF% (r = 0.51; p = 0.004). In patients who developed sepsis IPF% was inversely correlated with platelet count (r = ?0.60; p < 0.001) and had high values before sepsis became manifest, decreasing significantly on the 2nd day thereafter.

Conclusions

In patients without sepsis at ICU admission IPF% increases before sepsis becomes manifest. Measuring IPF% through an easily available technology can therefore provide an early cellular marker predicting the development of sepsis.  相似文献   

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