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1.
为研制用于预防金黄色葡萄球菌性奶牛乳房炎的免疫疫苗,使用4株血清型为外膜多糖336型(336PS)、荚膜多糖5型(CP5)和荚膜多糖8型(CP8)强毒力金黄色葡萄球菌试制出金黄色葡萄球菌多价疫苗,并通过抗体水平监测、攻毒保护试验以及两个牧场的田间试验对免疫效果进行了评价。抗体水平监测显示,首免后30 d即可产生较高的抗体水平,二次免疫后有效抗体水平可持续到产后210 d;人工攻毒保护试验显示,疫苗对金黄色葡萄球菌引起的临床型乳房炎的保护率为89%;田间试验结果表明,该疫苗对奶牛临床型乳房炎的保护率在75.2%~83.3%之间,对隐性乳房炎的保护率在49.9%~52.2%之间,免疫组日产奶量较对照组平均提高1.26 kg以上。上述结果表明,所试制的疫苗可以显著降低泌乳牛金黄色葡萄球菌引起的临床型和隐性乳房炎发病率并提高产奶量。  相似文献   

2.
奶牛乳房炎金黄色葡萄球菌荚膜多糖研究概况   总被引:1,自引:0,他引:1  
金黄色葡萄球菌是引起奶牛乳房炎的一种主要致病菌,其中94%~100%的金黄色葡萄球菌都含有荚膜多糖.荚膜多糖共分11个血清型(CP1~CP11),而5型和8型占临床分离株的70%~80%,为优势血清型;由于荚膜多糖的抗吞噬作用和黏附作用,决定了金黄色葡萄球菌的致病性和免疫原性.因此,基于荚膜多糖的疫苗可能是更有效的抗奶...  相似文献   

3.
为评价猪链球菌病-副猪嗜血杆菌病表面展示二联亚单位疫苗对小鼠的保护效果,本研究在应用融合PCR构建副猪嗜血杆菌(HPS)保护性抗原AfuA-OppA2和CdtB-OppA基因的串联序列基础上,将其分别插入pMD-28a-INP-His表面展示质粒,并将猪链球菌(SS)保护性抗原MRP、SLY基因也分别克隆至该表面展示质粒,构建pMD-INP-CdtB-OppA、pMD-INP-AfuA-OppA2、pMD-INP-MRP、pMD-INP-SLY 4种重组质粒,分别转入E.coli BL21(DE3)中诱导表达,使重组蛋白AfuA-OppA2、CdtB-OppA、MRP、SLY分别展示于E.coli BL21(DE3)的菌体表面。将灭活后的4种重组大肠杆菌按等质量比混合并添加ISA 201佐剂乳化制成表面展示二联亚单位疫苗,间隔14 d两次免疫KM小鼠,另设含rAfuA、rOppA2、rCdtB、rOppA、rMRP、rSLY的纯化蛋白亚单位疫苗免疫组、副猪嗜血杆菌-猪链球菌(HPS-SS)灭活疫苗免疫组和PBS对照组,二次免疫后的小鼠血清通过ELISA检测相应抗体和细胞因子水平。随后用HPS血清5型HN10株、血清13型ZD12株和SS血清2型M126株和血清9型GZ2株进行攻毒。结果显示:表面展示二联亚单位疫苗刺激小鼠产生的抗体和细胞因子水平与纯化蛋白亚单位疫苗组相当。用副猪嗜血杆菌5型和13型菌株分别攻毒后,表面展示二联亚单位疫苗对小鼠的保护率均为80%,其对HPS 5型菌株攻毒的小鼠保护率低于纯化蛋白亚单位疫苗(90%),但对HPS 13型菌株攻毒小鼠的保护率高于纯化蛋白亚单位疫苗(60%),且均优于HPS-SS灭活苗(80%和60%)。用SS 2型和9型菌株分别攻毒后,表面展示二联亚单位疫苗对小鼠的保护率为50%和80%,低于纯化蛋白亚单位疫苗的保护效果(100%和80%),优于HPS-SS灭活苗(50%和60%)。以上结果表明,基于表面展示技术的猪链球菌病-副猪嗜血杆菌病二联亚单位疫苗能刺激机体产生相应的抗体,其对HPS血清5型、13型和SS血清2型、9型菌株的攻击均能提供良好的交叉保护,保护效果优于HPS-SS灭活苗。本研究首次为基于表面展示技术的亚单位疫苗的研制提供实验依据,为细菌疫苗的研发提供新思路。  相似文献   

4.
A vaccine composed of three field isolates of Staphylococcus aureus (S. aureus) derived from cases of mastitis in cows was developed. The vaccine was administered to nine uninfected cows while 10 other cows were used as controls. All cows were challenged with a highly virulent S. aureus strain administered into two quarters of each cow. Quarters were tested for clinical signs, secretion of S. aureus, and somatic cell count (SCC). No systemic effects were observed in any of the cows, vaccinated or control. Vaccinated cows had 70% protection from infection compared with fewer than 10% in the controls. Moreover, all quarters challenged in the vaccinated cows, regardless of whether they were successfully infected or not with S. aureus, exhibited very mild inflammatory reactions, identified by their low SCCs (<100,000).  相似文献   

5.
ABSTRACT: Staphylococcus aureus is an important cause of nosocomial and community-acquired infections in humans and animals, as well as the cause of mastitis in dairy cattle. Vaccines aimed at preventing S. aureus infection in bovine mastitis have been studied for many years, but have so far been unsuccessful due to the complexity of the bacteria, and the lack of suitable vaccine delivery vehicles. The current study developed an Escherichia coli protein expression system that produced a recombinant staphylococcal enterotoxin A (rSEA) encapsulated into biodegradable microparticles generated by polylactic-co-glycolic acid (PLGA) dissolved in methylene chloride and stabilized with polyvinyl acetate. Antigen loading and surface properties of the microparticles were investigated to optimize particle preparation protocols. The prepared PLGA-rSEA microspheres had a diameter of approximately 5 μm with a smooth and regular surface. The immunogenicity of the PLGA-rSEA vaccine was assessed using mice as an animal model and showed that the vaccine induced a strong humoral immune response and increased the percent survival of challenged mice and bacterial clearance. Histological analysis showed moderate impairment caused by the pathogen upon challenge afforded by immunization with PLGA-rSEA microspheres. Antibody titer in the sera of mice immunized with PLGA-rSEA microparticles was higher than in vaccinated mice with rSEA. In conclusion, the PLGA-rSEA microparticle vaccine developed here could potentially be used as a vaccine against enterotoxigenic S. aureus.  相似文献   

6.
A gangrenous dermatitis model was developed in broiler chickens, in which birds previously vaccinated at 14 days of age with a bursal disease virus vaccine were challenged at 4 wk of age with various bacterial combinations with the combination of subcutaneous and intramuscular injection. Gangrenous dermatitis lesions were not produced in birds injected with one of the Staphylococcus aureus isolates, either alone or in combination with various Clostridium septicum isolates. Other S. aureus isolates produced significant levels of gangrenous dermatitis either alone or in combination with the same C. septicum isolates. These same C. septicum isolates when given alone did not produce gangrenous lesions. Data from this experiment show the highest level of mortality occurred in birds challenged with a mixture of C. septicum and S. aureus isolates, whereas lower or no mortality was associated with the same isolates given separately. The data clearly demonstrate that the pathogenicity of isolates responsible for gangrenous dermatitis varies widely, indicating that the frequency and severity of lesion production, as well as the occurrence of mortality, are largely dependent upon the specific isolate or isolates with which the birds are challenged.  相似文献   

7.
Daily injections of an anti-inflammatory milk-derived factor (MDF) into mice increased resistance to Staphylococcus aureus challenge, and reduced leukocyte infiltration. Intraperitoneal injection of MDF into lactating mice prior to S. aureus intramammary challenge resulted in greater milk secretory activity and less inflammation compared with untreated controls, but had little effect on the number of S. aureus recovered from mammary tissue. Infusion of MDF directly into mouse mammary glands prior to challenge reduced S. aureus recovered after challenge. Incubation of bovine mammary macrophages in medium supplemented with MDF enhanced phagocytosis of opsonized S. aureus. In addition, infusion of 5 mg MDF into uninfected bovine mammary glands 24 h prior to S. aureus challenge resulted in fewer infections (five of ten) than in control quarters (seven of nine). Repeated daily injections of 5 mg MDF into S. aureus-infected quarters increased the percent of mammary neutrophils and decreased the recovery of S. aureus, but did not eliminate infections. Intravenous injection of 8 g MDF into cows resulted in pronounced leukopenia while the accompanying effect on mammary leukocytes was less marked but followed a similar course. Results suggest that the use of MDF in mice enhanced resistance to experimental infection and was beneficial in maintaining mammary secretory activity and reducing inflammation after bacterial challenge. In the cow, MDF promoted phagocytosis in vitro and was effective against challenge when infused intramammarily.  相似文献   

8.
为调查新疆奶牛乳房炎金黄色葡萄球菌主要流行血清型,本研究采用玻板凝集和双重PCR方法对不同地区奶牛乳房炎乳样中分离的117株金黄色葡萄球菌进行血清分型.结果表明荚膜多糖5型占10.26%(12/117),8型占13.68%(16/117),336型占64.96%(76/117),未分型菌株占11.11 %(13/117).该研究为新疆奶牛乳房炎金黄色葡萄球菌疫苗菌株的筛选提供了依据.  相似文献   

9.
The surface glycoprotein G is considered as the major neutralizing and protective antigen of bovine ephemeral fever virus (BEFV). Comparison of the deduced amino acid sequence of G protein of BEFV isolates during the period 1984-2004 outbreaks in Taiwan showed amino acid substitutions in the neutralizing epitopes. All the isolates differ markedly in the neutralizing epitope at the same amino acid positions compared to the currently available killed vaccine strain (Tn73). Tn88128 strain isolated in 1999 showed the maximum variability of 12 amino acids, 5 amino acid in the neutralization epitope and 7 apart from, respectively. Combinations of both Tn88128 (1999) and commercially available vaccine strain (Tn73) were developed and its safety was evaluated in mice, guinea pigs, calves, and pregnant cows. None of the animals showed any adverse effect or clinical signs. Calves were immunized with commercial vaccine (Tn73) and, combined vaccine (Tn73 and Tn88128), respectively, with adjuvants such as Al-gel and water-in-oil-in-water (w/o/w) oil and PBS alone and challenged with Tn88128 strains. Except PBS administered animals, all the vaccinated animals showed protective immune response. However, animals immunized with combined vaccine plus w/o/w adjuvant elicited stronger neutralization antibodies and long lasting immunity compared to other vaccines.  相似文献   

10.
Staphylococcus aureus is a common pathogen which can colonise and infect not only man, but also domestic animals. Especially, infection of cattle is of high economic relevance as S. aureus is an important causal agent of bovine mastitis. In the present contribution, a DNA microarray was applied for the study of 144 different gene targets, including resistance genes and genes encoding exotoxins, in S. aureus isolated from cows. One hundred and twenty-eight isolates from Germany and Switzerland were tested. These isolates were assigned to 20 different strains and nine clonal complexes. The majority of isolates belonged either to apparently closely related clonal complexes 8, 25, and 97 (together 34.4%) or were related to the sequenced bovine strain RF122 (48.4%). Notable characteristics of S. aureus of bovine origin are the carriage of intact haemolysin beta (in 82% of isolates tested), the absence of staphylokinase (in 89.1%), the presence of allelic variants of several exotoxins such as toxic shock syndrome toxin and enterotoxin N, and the occurrence of the leukocidin lukF-P83/lukM (in 53.1%). Two isolates were methicillin-resistant S. aureus (MRSA). One of them was a clonal complex 8 MRSA related to the epidemic MRSA strain Irish 01. The other one belonged to ST398/spa-type 34 resembling a newly emerging MRSA strain which has been described to occur in humans as well as in domestic animals. The presence of these two strains highlights the possibility of transfers of S. aureus strains between different host species.  相似文献   

11.
Mice immunized with an aromatic-dependent (aro-) S. dublin strain CS101 by either the intraperitoneal (i.p.) or oral route, were protected against oral challenge with a virulent S. dublin strain CS90, the degree of protection being the greatest when mice had received 3 immunizing doses at weekly intervals. Mice immunized with an aromatic-dependent (aro-) S. typhimurium strain CS332 by the i.p. or oral routes were protected against challenge with virulent S. dublin strain CS90 at 1 or 2 weeks but not at 3 or 4 weeks post-immunization. Mice immunized with 1 dose of aro- S. dublin strain CS101 by the i.p. route developed low levels of lipopolysaccharide (LPS) and flagellin-specific antibody but no delayed-type hypersensitivity (DTH) whereas those immunized with 2 or 3 doses developed significantly higher antibody titres and DTH. In contrast, mice immunized by the oral route developed neither significant antibody response nor DTH. The aro- S. dublin strain CS101 could not be detected beyond day 28 post-inoculation in visceral organs including liver, spleen, mesentery, small intestine, caecum or large intestine of mice inoculated by the i.p. route or in mice inoculated by the oral route with the exception of day 42 post-inoculation. Challenge of mice previously immunized with 3 doses of the aro- S. dublin strain CS101 by the i.p. or oral route with virulent S. dublin strain CS90 resulted in their rapid clearance from the above visceral organs. Calves immunized with the aro- S. dublin strain CS101 by either the intramuscular (i.m.) or oral routes were significantly protected against oral challenge with virulent S. dublin strain CS90. In contrast to the observations in mice, somatic (O) and flagellar (H) antibody titres of calves immunized by either route were negligible as were anti-LPS antibody titres. However, flagellin-specific antibody titres were higher in calves immunized by the i.m. than the oral route. These results indicate that the protection observed in immunized mice or calves against oral challenge with virulent S. dublin was unlikely to have been mediated by humoral salmonella-specific immune mechanism(s).  相似文献   

12.
Staphylococcus (S.) aureus is a major udder pathogen causing bovine mastitis. Some pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), enhance extracellular and intracellular growth of S. aureus, indicating that the inflammatory process favors S. aureus infection. Helenalin is a sesquiterpene lactone with potent anti-inflammatory properties. This study was designed to evaluate the effects of helenalin on S. aureus infection. First, in vitro experiments were conducted. These studies revealed that proliferation of S. aureus in bovine mammary epithelial MAC-T cells treated in the presence or absence of TNF-alpha was markedly reduced in the presence of helenalin. Secondly, in vivo effects of helenalin were investigated. Lactating mice treated in the presence or absence of helenalin were challenged by the intramammary route with S. aureus and the bacteria in the mammary glands were counted 12 h after infection. Significantly less numbers of bacteria were recovered from the infected glands of helenalin-treated mice compared with untreated mice. Moreover, histological examination of mammary tissue from helenalin-treated mice that were challenged with S. aureus indicated that helenalin is able to significantly reduce leukocyte infiltration in the mammary gland following S. aureus inoculation. Our results show that helenalin reduces S. aureus intracellular growth and experimental S. aureus infection. We conclude that helenalin may be of potential interest in the treatment of S. aureus-induced mastitis in the bovine species.  相似文献   

13.
The expression of inflammatory cytokines, including interleukin (IL)-6, IL-8, IL-12, granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma, by milk somatic cells was characterized by real-time polymerase chain reaction (PCR) in dairy cows experimentally challenged with either E. coli (n = 8) or S. aureus (n = 8). The mRNA abundance of a target gene was calibrated with that of a reference gene (beta-actin) and expressed as fold of induction over the control quarter at each time point. At no single time point did all eight quarters challenged with the same type of bacteria demonstrated increased expression of a target gene and there was large variation among animals at each given time. As a consequence, most tested comparisons were not statistically significant except the peak time points of IL-8 expression (75- and 29- fold in glands challenged with E. coli and S. aureus, respectively). However, the average fold induction of all targeted cytokines was increased in response to both bacterial challenges with the exception of IFN-gamma. The expression of IFN-gamma was only increased in milk somatic cells isolated from E. coli, but not S. aureus, challenged mammary glands. Moreover, upregulated expression of cytokine genes had higher magnitudes and/or faster responses in glands challenged with E. coli in comparison with those challenged with S. aureus. We propose that the compromised upregulation of inflammatory cytokines in S. aureus infected glands may, at least partially, contribute to the chronic course of infection caused by this pathogen. Further research on identifying factors responsible for the differentially expressed cytokine profiles may be fundamental to developing strategies that mitigate the outcome of bovine mastitis.  相似文献   

14.
Mice and swine immunized subcutaneously with live vaccine prepared from acriflavine-fast attenuated Erysipelothrix rhusiopathiae, strain Koganei (serotype 2), were challenge exposed to virulent strains of E rhusiopathiae of various serotypes. Vaccinated mice did not die after challenge exposure to serotypes 1a, 1b, 2, 3, 5, 6, 7, 8, 9, 11, 12, 15, 16, 18, 19, 21, or N, but 20% to 30% mortality occurred in vaccinated mice challenge exposed to serotypes 10, 14, 20, or 22. Nonvaccinated control mice died after challenge exposure to all serotypes tested. Vaccinated swine challenge exposed to strain 14B (serotype 9) or strain 2179 (serotype 10) developed localized urticarial lesions at the site of intradermal exposure. Vaccinated swine challenge exposed to serotypes 1a, 1b, 2, 5, 8, 11, 12, 18, 19, or 21 did not have clinical signs of acute erysipelas. Nonvaccinated control swine developed acute generalized erysipelas or localized lesions at the site of intradermal exposure.  相似文献   

15.
Wang LH  Li XY  Jin LJ  You JS  Zhou Y  Li SY  Xu YP 《Veterinary microbiology》2011,149(3-4):415-421
The objective of this in vitro study was to evaluate the potential of egg yolk immunoglobulins (IgYs) for treating mastitis caused by Staphylococcus aureus. Specific IgY against type 5 (IgY-T5), type 8 (IgY-T8) and type 336 (IgY-T336) S. aureus strains were obtained by immunizing hens with whole cell vaccines and the IgY produced were then purified to around 80% purity using a water dilution method coupled with salting out and ultra-filtration. Enzyme-linked immunosorbent assay indicated that the IgY specifically targeted the three homologous strains. A growth inhibition assay was performed in Columbia broth (non-encapsulated form) and phosphate-buffered saline (encapsulated form) for an 8h incubation. The results showed that IgY-T336 significantly inhibited (but only 1.5 log units; P<0.01) the growth of all three strains at 15 mg/ml in the Columbia broth. In contrast, the same concentrations of IgY-T5 and IgY-T8 did not show obvious bacteriostatic activity against the two homologous strains. In phosphate buffered saline, no inhibition of the two encapsulated strains was observed with IgY-T5, IgY-T8 and IgY-T336. However, IgY-T336 reduced live bacteria by 1.0 log unit against strain 336 compared with the control. An internalization test indicated that all of the specific IgY (at 5mg/ml) significantly (about 3.0 log units of the control; P<0.01) blocked the internalization of their homologous strains by bovine mammary epithelial cells (MAC-T cells) within 6h. These results suggested that research on the application of IgY as a treatment for mastitis caused by S. aureus should be focused on the internalization inhibition activity rather than on the growth inhibition activity of the IgY.  相似文献   

16.
The generation of aromatic-dependent (aro-) Salmonella havana (Group G2, 01, 13, 23) from a smooth wild-type parent strain by transduction with phage P1 is reported. Mice immunized with this live aro- S. havana strain (CS234) by the intraperitoneal (i.p.) route were protected against challenge with wild-type S. havana, whereas those immunized by the oral route were not. Mice immunized with two doses of formalin-killed aro- S. havana by the i.p. route were also unprotected, in spite of high antibody titers. However, only those mice immunized with live aro- S. havana by the i.p. route developed significant delayed-type hypersensitivity. Following i.p. inoculation in mice, the aro- S. havana strain CS234 was detected in the liver, spleen and mesenteric lymph nodes on day 9 but not on day 15 post-inoculation (p.i.). On the other hand, when mice were inoculated with the parent wild-type strain (CS4) or the aro- derivative strain CS234 by the oral route, the organisms were recovered from the mesenteric lymph nodes and intestine only on day 3 but not on day 6 post-inoculation. In sheep inoculated with the aro- strain CS234 in the gastroc muscle, organisms were recovered from the muscle, and popliteal and medial iliac lymph nodes for up to 21 but not 28 days p.i. However, no mutant organisms were recovered from liver, spleen, mesenteric lymph nodes or faeces. In orally-inoculated sheep, the mutant organisms were recovered from the mesenteric lymph nodes, rumen, intestinal contents, and faeces up to 14-21 days post-inoculation but not at 28 days. When sheep immunised with the aro- S. havana strain CS234 by the intramuscular or oral route were challenged with the parent wild-type S. havana strain CS4 by the oral route, the latter strain was detectable in the mesenteric lymph nodes and faeces of immune sheep up to 14 days post-challenge in contrast with the non-immune sheep, where the challenge strain was detectable even at 28 days post-challenge. Only sheep immunized by the intramuscular route developed high antibody levels and delayed-type hypersensitivity.  相似文献   

17.
Detection of capsular polysaccharide (CP) in milk of cows with natural intramammary infection caused by Staphylococcus aureus was attempted. Five quarters of 5 cows harboring S aureus strains that produce type-8 CP were selected. Using an ELISA with a monoclonal antibody, type-8 CP was not detected in extracts prepared from fresh milk collected aseptically. By contrast, CP was easily detectable after incubation of infected milk at 38 C for 20 hours. Quantitation of CP in extracts from incubated milk samples by use of ELISA indicated a great variation of CP expression by strains. Although an incubation step was necessary to detect CP, results of the study indicate that CP may be expressed in vivo during intramammary infection caused by S aureus.  相似文献   

18.
BALB/c mice were immunized with live rough Brucella abortus RB51 or smooth Brucella melitensis Rev. 1 vaccines and challenged with a B. melitensis field strain. Protection was assessed by a variety of serological tests and recovery of vaccinal and challenge strains by culture. Mice vaccinated with RB51 gave negative results in the conventional serological tests prior to challenge, namely; standard tube agglutination test (SAT), Rose Bengal plate test (RBPT), buffered acidified plate antigen test (BAPAT), and mercaptoethanol test (MET). Sero-conversion took place to a whole-cell bacterial buffered RB51 antigen after vaccination and persisted for 7 weeks post-vaccination. Mice challenged with B. melitensis were assessed for bacterial load and immune response for 12 weeks after challenge. Protection units were showed that Rev. 1 vaccine was superior to RB51 vaccine in protection of mice against B. melitensis. However, RB51 vaccine has the advantage that it would not elicit antibodies to standard serological tests based on the LPS O antigen. RB51 vaccine could therefore be used for control of B. melitensis infection and avoid confusion in the use of standard sero-diagnostic tests.  相似文献   

19.
为鉴定一株从雁鸭脏器内分离到的革兰氏阴性细菌,本研究对该菌进行分离培养、细菌16S rRNA序列比对、动物试验和药物敏感性试验。结果表明该分离菌与多杀性巴氏杆菌(P.mutocida)(AF224297)同源性达99.88%,毒力强,能够致死家兔、小鼠和鸡,对氧氟沙星等药物敏感。分离菌的荚膜抗原血清型特异性基因PCR产物与荚膜血清A型P.mutocida hyaD-hyaC基因同源性达99.9%,确定该菌株为荚膜血清A型P.mutocida。本研究首次从雁鸭体内分离到A型P.mutocida。  相似文献   

20.
In an attempt to protect pigs against swine pleuropneumonia induced by Actinobacillus pleuropneumoniae (SPAP) by neutralizing the effects of three virulence factors of A. pleuropneumoniae--the capsular polysaccharide (CP), the lipopolysaccharide (LPS), and the hemolysin protein (HP)--two subunit conjugate vaccines were prepared by covalently coupling the CP to the HP and the LPS to the HP. The CP, LPS, and HP were isolated from A. pleuropneumoniae, strain 4074, serotype 1, and the protective efficacy of the conjugate vaccines in swine experimentally infected with A. pleuropneumoniae was evaluated. Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP, LPS, and HP was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs. Thus, on the basis of mortality, weight gains, and pulmonary lesion formation, the two conjugate vaccines used in conjunction with one another provide noticeable protective efficacy against SPAP.  相似文献   

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