朱顶红无菌苗叶片高效再生体系

以朱顶红（Hippeastrum vittatum）叶片为外植体进行离体培养，具有取材方便、试材充足、成本低等优势，但叶片诱导再生率极低，是朱顶红离体培养的一大难题。本试验中分别以‘花孔雀’和‘黑天鹅’朱顶红无菌苗叶片为外植体，探究了不同植物生长调节剂和不同取材部位对不定芽诱导和继代增殖的影响。结果表明：最佳外植体为 MS 培养基中培养 10 d 形成的幼嫩叶片基部（0.5 cm），在光照 16 h · d-1（光照强度 36 μmol · m-2 · s-1）下，不定芽诱导的最适培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 2 mg · L-1 TDZ，两个品种的不定芽均以间接途径发生，其中‘花孔雀’在培养 40 d 后形成愈伤组织，55 d形成不定芽，诱导率可达 69.44%；‘黑天鹅’在培养 45 d 后形成愈伤组织，65 d 形成不定芽，诱导率达到 66.67%；最适体细胞胚诱导培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 PIC，‘花孔雀’和‘黑天鹅’的诱导率分别达到 66.67%和 63.89%；最佳不定芽增殖培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 1 mg · L-1 TDZ，‘花孔雀’和‘黑天鹅’的增殖系数分别达到 4.67 和 3.46；在不添加植物生长调节剂的 MS培养基中进行生根培养，30 d 后两个品种的生根率均达到 100%；将生根培养 30 d 的小植株转移至室温条件下放置 3 d，摘去封口膜再驯化 3 d 后，移栽至经高温消毒的草炭︰蛭石（体积比）为 1︰1 的基质中，成活率达到 100%     

Characterizing the Serum Proteome of Donkeys (Equus asinus)

Serum and plasma are commonly used in clinical practice considering the widely accepted fact that the “normal” protein expression pattern of a healthy animal changes under disease conditions. We herein used a label-free mass spectrometry–based quantitative proteomics approach to characterize the serum proteome of donkeys. A total of 277 unique proteins were identified from 2,388 unique peptides. Gene ontology analyses showed that the most frequent processes were related to metabolic activities and biological regulation, response to stimulus, and immune system processes. The main annotated areas of origin were the extracellular region, extracellular region part, and organelle, and their molecular functions included binding, catalytic activity, and molecular function regulator. Analyses using the Clusters of Orthologous Groups for Eukaryotic Complete Genomes database indicated that the identified proteins could be categorized into three main groups: signal transduction mechanisms, amino acid transport and metabolism, and defense mechanisms. Most of the unique proteins were associated with the complement and coagulation cascades, and they participated in several disease-related metabolic pathways. Our results should be crucial for further analyses of changes in different physiological and pathophysiological conditions in donkeys.     

Acute phase response in mice experimentally infected with Trypanosoma congolense: a molecular gauge of parasite-host interaction

Mice infected with Trypanosoma congolense developed a severe anaemia 1 week after infection, which persisted till treatment with diminazine aceturate when the packed cell volume (PCV) recovered to pre-infection levels. This was accompanied by a marked increase in the plasma levels of the acute phase proteins (APP), serum amyloid P-component (SAP) and haptoglobin (Hp). The initial peak levels of Hp and SAP were attained 7 and 12 days post-infection (DPI), respectively. Thereafter SAP levels decreased significantly to near pre-infection levels, but later increased even after treatment to give a second peak 34 DPI after which there was a decline till the study was terminated. The Hp levels on the other hand decreased to an intermediate level after the initial peak increasing to a second peak 22 DPI. Thereafter Hp decreased significantly following diminazine aceturate treatment to reach pre-infection levels within 5 days post-treatment. This indicates that T. congolense-infected mice develop severe anaemia accompanied by an acute phase response leading to an increase in SAP and Hp but that following treatment divergent responses occurred indicating differences in the pathways for stimulation of the APP. Haptoglobin was shown to be an earlier indicator of infection and a better marker in monitoring the response to treatment.     

Renal amyloidosis in cattle with inflammatory diseases

BACKGROUND: The association of inflammatory diseases such as traumatic reticuloperitonitis (TRP), mastitis, metritis, and pododermatitis with renal amyloidosis in cattle is poorly described. HYPOTHESIS: Serum amyloid A (SAA) levels are elevated during inflammatory diseases, and renal amyloidosis is formed as a complication. ANIMALS: This study was conducted with 82 crossbred cattle with mastitis (n = 18 cows), metritis (n = 11 cows), TRP (n = 30 cows), and pododermatitis (n = 23 : 15 cows and 8 beef cattle). Ten clinically healthy cows served as controls. Methods: Hematological, urinary, and blood parameters, including SAA, were measured by an automated procedure provided with trade kits. Determination of amyloidal structures was made by histopathological examination of renal biopsy specimens. RESULTS: At the end of this trial, amyloidosis was detected in 5 cows displaying typical nephrotic syndrome, with hypoproteinemia and proteinuria in combination with polyuria and weight loss. Furthermore, it was observed that cows with renal amyloidosis had significantly higher (P < .01) total leukocyte counts, serum and urine enzyme activities, and urea and creatinine concentrations, with lower serum total protein concentrations, when compared with animals without renal amyloidosis. CONCLUSIONS AND CLINICAL IMPORTANCE: The incidence of AA amyloidosis in cattle in this study suggests that cattle with mastitis, metritis, and pododermatitis have a high prevalence of systemic amyloidosis in response to inflammation.     

In vivo and in vitro efficacy of zoledronate for treating oral squamous cell carcinoma in cats

BACKGROUND: Feline oral squamous cell carcinoma (OSCC) may cause painful bone destruction. Given the local invasiveness and rapid clinical progression of OSCC, conventional therapies are often palliative. In human cancer patients, zoledronate exerts anticancer effects by inhibiting tumor-induced angiogenesis and malignant osteolysis. HYPOTHESIS: Zoledronate will exert in vitro and in vivo anti-angiogenic and antiresorptive effects in feline OSCC. ANIMALS: Eight cats with OSCC were prospectively treated with zoledronate and conventional treatment modalities. METHODS: In vitro, zoledronate's effects in modulating soluble vascular endothelial growth factor (VEGF) secretion and receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) expression were investigated in a feline OSCC cell line (SCCF1). In vivo, basal serum C-telopeptide (CTx) concentrations were compared among normal and OSCC-bearing cats, and the biologic effects of zoledronate administration in cats with naturally occurring OSCC were quantified by serially assessing circulating serum VEGF and CTx concentrations. RESULTS: In vitro, zoledronate concentrations greater than 3 microM reduce soluble VEGF secretion in the SCCF1 cell line. The expression of RANKL in the SCCF1 cell line was also modulated by zoledronate, with low concentrations (3 microM) decreasing but higher concentrations (30 microM) increasing RANKL expression in comparison with untreated cells. In vivo, cats with bone-invasive OSCC had greater serum CTx concentrations in comparison with geriatric, healthy controls. Treatment with zoledronate rapidly decreased circulating serum VEGF and CTx concentrations in cats with spontaneously occurring OSCC. CONCLUSIONS AND CLINICAL IMPORTANCE: Zoledronate exerts in vitro and in vivo effects that may favor the slowing of tumor growth and pathologic bone turnover associated with OSCC.     

肉鸡大肠杆菌的分离鉴定

从豫、鲁、冀接壤地区部分肉鸡场送检的病料中分离出49株大肠杆菌,确定42株菌分属11个血清型,其中02、078、01、074、011为主要血清型,分别占分离株的26.2%,19.0%,11.9%,9.5%和9.5%,表明该地区血清型复杂.药敏试验表明,所分离的49株大肠杆菌对丁胺卡那霉素、氟本尼考、头孢噻呋最敏感,高敏菌株达94%～100%,可作为防治肉鸡大肠杆菌病的首选药物.     

不同营养添加剂对热应激异育银鲫血液生化指标的影响

为探讨VC、VE和小肽对鱼类抗热应激能力的影响及营养调控作用。本研究选择体重(100±3)g的异育银鲫150尾,随机分成5组,即Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组。Ⅰ为对照组,饲喂基础日粮,Ⅱ、Ⅲ、Ⅳ和Ⅴ组为试验组,分别饲喂在基础日粮中添加VC制剂100mg/kg、VE制剂100mg/kg、VC制剂100mg/kg VE制剂100mg/kg和1%小肽(替代基础日粮中等量鱼粉)的日粮。常温(25℃)下饲养42d后,将鱼在31℃水中应激24h,随机抽取应激前后3尾鱼血样,测定应激前后血清超氧化物歧化酶(SOD)、乙酰胆碱脂酶(TchE)、谷草转氨酶(GOT)活性。结果表明:在常温养殖条件下,VE组血清TchE活性显著降低(P<0.05),GOT活性显著升高(P<0.05),其他各组鱼血清3种酶活性与对照组相比差异不显著(P>0.05);热应激后,与常温状态相比,添加VC、VE使得SOD活性降低,GOT、TchE活性升高;VC与VE协同作用引起TchE、GOT活性显著上升(P<0.05),SOD活性没有变化;添加小肽能使TchE活性显著升高(P<0.05),SOD、GOT活性不变。与高温对照组相比,VC与VE协同作用组显著提高了SOD、TchE活性(P<0.05);添加小肽可提高SOD活性,降低TchE和GOT活性。由此表明,通过在日粮中补充抗热应激营养添加剂,有助于增强鱼类机体的抗热应激能力;添加小肽以及VC与VE协同作用对缓解异育银鲫热应激损伤具有良好的调控作用。     

Effect of serum amyloid A on invasion,migration and MAPK signaling pathway in osteosarcoma cells

AIM: To investigate the effect of silencing of serum amyloid A (SAA) on the viability, apoptosis, migration and mitogen-activated protein kinase (MAPK) signaling pathway in osteosarcoma U2OS cells. METHODS: Small interfering RNA (siRNA) targeting SAA was transfected into U2OS cells to silence the expression of SAA gene. The U2OS cells were divided into blank control group, negative control group, and experimental group. The cells in negative control group and experimental group were transfected into negative control siRNA and SAA-siRNA, respectively. The cells in blank control group were without any treatment. The viability of the cells was measured by MTT assay and the apoptotic rate was analyzed by flow cytometry with Annexin V-FITC/PI double staining. The migration and invasion abilities of the cells were detected by Transwell chamber assay. The protein levels of SAA, phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated c-Jun N-terminal kinase (p-JNK) in the cells were determined by Western blot. RESULTS: The protein expression of SAA in SAA-siRNA group was significantly lower than that in blank control group (P<0.05). Compared with blank control group, the cell viability in SAA-siRNA group was significantly decreased (P<0.05), the apoptotic rate was significantly increased (P<0.05), and the invasion and migration abilities were significantly decreased (P<0.05). The protein levels of p-p38 MAPK and p-JNK in SAA-siRNA group were significantly lower than those in blank control group (P<0.05), and no significant difference of total JNK and p38 protein levels was observed. CONCLUSION: Silencing of SAA expression inhibits the viability of osteosarcoma cells, induces apoptosis and decreases the migration of osteosarcoma cells, which may be related to the activation of MAPK signaling pathway.     

蛋氨酸锌对40日龄吉戎兔血清生化指标的影响

随机选取40日龄健康吉绒Ⅱ系吉戎兔72只,分成4组,每组3个重复,每个重复6只。试验I为对照组,饲喂基础饲料;试验Ⅱ、Ⅲ、Ⅳ分别添加40,80,120mg/kg蛋氨酸锌。结果表明,日增重在试验期0～15d期间及16～30d期间,试验Ⅲ与其它组差异显著（P〈0．05）,分别比对照组、试验Ⅱ、Ⅳ组提高13．94％和11．31％、6．60％和5．93％及7．47％和8．44％。料重比在试验期0～15d期间,试验Ⅲ与其它组差异显著（P〈0．05）,分别比对照组、试验Ⅱ、Ⅳ组改善12．23％、6．19％及6．95％;在试验期16～30d期间,试验Ⅲ和试验Ⅱ差异不显著（P〉0．05）,但与对照组和试验Ⅳ差异显著（P〈0．05）,试验Ⅲ比对照组、试验Ⅱ、Ⅳ组改善10．16％、5．60％、7．78％。试验Ⅳ血清锌和白蛋白含量与对照组差异显著（P〈0．05）。试验组血清碱性磷酸酶、总蛋白含量与对照组差异显著（P〈0．05）,各处理组间尿素氮差异不显著（P〉0．05）。结论为在40日龄吉戎兔饲粮中添加80mg/kg蛋氨酸锌适宜。     

Evaluation of a serum-based PCR assay for the diagnosis of canine monocytic ehrlichiosis

The aim of this study was to estimate the relative diagnostic sensitivity and specificity of a polymerase chain reaction (PCR) assay in the serum of dogs with naturally occurring non-myelosuppressive canine monocytic ehrlichiosis (CME), and to investigate the association between PCR positivity and immunofluorescence antibody (IFA) titres for Ehrlichia canis. Serum samples obtained from 38 dogs with non-myelosuppressive CME and 12 healthy dogs were analyzed retrospectively. Each serum sample was analyzed in triplicate using an E. canis-specific nested PCR assay targeting a 389 bp sequence of the 16S rRNA gene. E. canis DNA was amplified in 24 of 38 (63.1%) affected dogs; all samples from healthy dogs were negative. A high level of agreement was found among the PCR replicates (P < 0.0001). Median IFA titre of the 24 PCR-positive dogs was significantly lower than that of the PCR-negative infected dogs (P = 0.0029), indicating that E. canis DNA may circulate prior to the development of a high antibody titre. Serum-based PCR analysis is suggested for the early diagnosis of CME when whole blood samples are not available.