滴度及注射方式对2型腺相关病毒载体表达效果的影响

由于腺相关病毒载体(adeno-associated viral vector,AAV)本身的侵染不具有神经元特异性,其在神经系统相关研究中会存在侵染范围不符合试验要求的情况,因此本试验拟研究不同滴度和注射方式对病毒侵染范围的影响。结果显示,在注射较高滴度的AAV2-CMV-EGFP(1.3×10¹¹mg/L)3周后,其侵染范围可由延髓中缝苍白核(raphe pallidus nucleus,RPa)区域延伸至下丘脑室旁核(paraventricular nucleus,PVN)区域;较低滴度(1.3×10⁸mg/L)结合压力注射的方式可以在一定程度上限制AAV的侵染范围,并且载体携带的EGFP荧光基团在637 nm波段不具有明显的假阳性信号。     

Control of insect pests using trunk injection in a newly established apple orchard

We evaluated the potential for using preplant trunk injections of emamectin benzoate in nonbearing apple trees. Trees were evaluated for pest injury and emamectin residues throughout the planting season and into the following year. Injections into the trunk best delivered emamectin benzoate to the canopy compared with injections into the taproot, and the higher rate reduced insect pests more than the lower rate. In the following year, differences in insect control between trunk and root injections were less pronounced, but the higher rate of emamectin benzoate persisted longer and better reduced pests relative to the other treatments.     

Embryo-mediated genome editing for accelerated genetic improvement of livestock

Selecting beneficial DNA variants is the main goal of animal breeding. However, this process is inherently inefficient because each animal only carries a fraction of all desirable variants. Genome editing technology with its ability to directly introduce beneficial sequence variants offers new opportunities to modernize animal breeding by overcoming this biological limitation and accelerating genetic gains. To realize rapid genetic gain, precise edits need to be introduced into genomically-selected embryos, which minimizes the genetic lag. However, embryo-mediated precision editing by homology-directed repair (HDR) mechanisms is currently an inefficient process that often produces mosaic embryos and greatly limits the numbers of available edited embryos. This review provides a summary of genome editing in bovine embryos and proposes an embryo-mediated accelerated breeding scheme that overcomes the present efficiency limitations of HDR editing in bovine embryos. It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with elite edited blastomeres or embryonic pluripotent stem cells to resolve mosaicism, enable multiplex editing and multiply rare elite genotypes. Such a breeding strategy would enable a more targeted, accelerated approach for livestock improvement that allows stacking of beneficial variants, even including novel traits from outside the breeding population, in the most recent elite genetic background, essentially within a single generation.     

硫酸卡那霉素注射液含量测定的不确定度分析

为研究微生物检定法(管碟法)测定硫酸卡那霉素注射液生物效价试验中各因素对结果的影响,采用不确定度分析,将影响因素归纳为称量、稀释、加样、测定、平行试验,量化各不确定度分量,评估各因素的影响因子。结果表明,牛津杯加样体积对测定结果影响最大,提示试验中应尽可能精确控制加样体积。     

Protective effect of Huangqi injection combined with puerarin injection on myocardium of type 2 diabetes mice

AIM:To investigate the effect of Huangqi injection combined with puerarin injection on the myocardium of the mice with type 2 diabetes. METHODS:Diabetic KKAy mice were randomly divided into model group and treatment group (Huangqi injection combined with puerarin injection). The male KKAy mice of the same age were used as control group. All mice were sacrificed at 21, 24 and 28 weeks. Morphological changes of the myocardium were observed by HE staining. Apoptosis of the cardiomyocytes was measured by TUNEL staining. The mRNA levels of glucose-regulated protein 78(GRP78), C/EBP hoinologous protein (CHOP) and p53 up-regulated modulator of apoptosis (PUMA) were detected by real-time PCR, and the protein levels of GRP78, CHOP, PUMA, caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, poly(ADP-ribose) polymerase (PARP) and cleaved PARP were determined by Western blot. RESULTS:Cardiomyocyte hypertrophy, partly dissolved sarcoplasm and necrosis were observed in model group, and these lesion were alleviated in treatment group. Obvious increased apoptosis in model group and significantly decreased apoptosis of cardiomyocytes in treatment group was observed (P<0.05). At 21, 24 and 28 weeks, the mRNA and protein levels of GRP78, CHOP and PUMA and the protein levels of cleaved caspase-3, cleaved caspase-9 and cleaved PARP in model group were increased significantly as compared with control group (P<0.01), and these in treated group were decreased compared with model group. CONCLUSION:Huangqi injection combined with puerarin injection has cardioprotective effects on type 2 diabetes mice and its mechanism of the action was implemented via inhibiting the activation of endoplasmic reticulum stress and caspase pathway, thus resulting in suppressed apoptosis.     

Clinical Therapeutic Effect of Praziquantel Injection on Buffalo Schistosomiasis

To observe the clinical therapeutic effect of 30% praziquantel injection on buffalo schistosomiasis, the sick buffalos naturally infected with Schistosoma japonicum were selected by miracidium hatching method. In experimental clinical trials, sick buffalos were randomly divided into five groups, high-dose (20 mg/kg), middle-dose (10 mg/kg) and low-dose (5 mg/kg) praziquantel injection groups, praziquantel tablet group (30 mg/kg) and positive control group. After the treatment of 30 d, the negative conversion rate of miracidium were 100.0%, 100.0%, 77.8% and 85.7%, respectively, in praziquantel injection groups with the high, middle and low dose and oral medication group. In expanded clinical trials, the sick buffalos were randomly divided into two groups, praziquantel injection group (10 mg/kg, i.m.) and praziquantel tablet group (30 mg/kg). The results showed that, the negative conversion rate of miracidium were all 100% in the former 52 patients and the latter 6 patients after 30 d. The research confirmed that praziquantel injection was significantly effective in the treatment of buffalo schistosomiasis and convenient for administration. It was concluded that 30% praziquantel injection could replace the traditional oral praziquantel tablet for the treatment of buffalo schistosomiasis, and the recommended dose was 10 mg/kg.     

Content Determination of Ivermectin in the Ivermectin Microemulsion Injection

In order to establish the determination method of ivermectin (IVM) in the ivermectin microemulsion injection by high performance liquid chromatography (HPLC), Hypersil ODS2 colunm (5 μm,4.6 mm×250 mm) was used in this study. The mobile phase composed of methanol, acetonitrile and water (35:60:5,V/V/V) at a flow rate of 1 mL/min. The detection wave length was set at 244 nm and the column temperature was 30 ℃. The results showed that the HPLC system suitability of IVM was good. A good linear correlation of IVM was observed within the concentration range 80 to 320 μg/mL, and the average recovery rate was 101.90%±2.94% with RSD was 2.88%, the regression equation was Y=22 700X+2 510 (R²=0.9998). The RSD of IVM content in ivermectin microemulsion injection was 1.86%. The method was accurate and reliable,reproducible,easy to operate, which could be used in new type of ivermectin microemulsion injection. The method could be used as the basis of quality control and establishing a quality standard, and to provide basis for quality evaluation, also can provide reliable reference for safe veterinary clinical application in the future.     

Establishment of Detection Methods for Dexamethasone Sodium Phosphate in Injections by UPLC-PDA

A detection method of dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection,and asragulus polysacharin injection was established by UPLC-PDA.The Waters ACQUITY UPLC^® BEH C18 column(2.1 mm×50 mm,1.7 μm) was used,the mobile phase consisted of 0.75% triethylamine phosphatic liquor/solution (pH 3.0)-methyl alcohol-acetonitrile(50:45:5),the flow rate was 0.3 mL/min,the column temperature had remained 25℃,photodiode array detector wavelength range was 190 to 400 nm,the recorded wavelength was 242 nm,the injection volume was 10 μL.The linear range of dexamethasone sodium phosphatel was 10.0 to 200.0 mg/L (R²=0.9999),the mean recovery and RSD for dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection and asragulus polysacharin injection were 97.54% and 1.14%,101.59% and 0.19%,100.92% and 0.92%,99.82% and 0.73%,100.08% and 0.62%,respectively.This method was accurate,easy and fast,it was suitable for detection of dexamethasone sodium phosphatein houttuynia injection and other four kinds of injections.     

参麦与黄芪注射液联用治疗卡氏肺孢子虫肺炎

为研究参麦与黄芪注射液联用对大鼠卡氏肺孢子虫肺炎(PCP)的治疗效果,以地塞米松连续肌肉注射Wistar大鼠6周,建立PCP大鼠模型,用参麦与黄芪注射液皮下注射治疗患病大鼠,同时设复方磺胺甲恶唑(SMZ-TMP)治疗对照组和PCP模型阳性对照组。通过各组大鼠基本情况变化、肺组织印片检查包囊、肺组织病理切片染色观察组织变化,探讨其治疗效果。结果发现,参麦与黄芪注射液联用治疗后,大鼠饮食、饮水增加,活跃程度恢复,体重不同程度增加,肺印片Pc包囊数量比治疗前明显减少,肺组织炎症反应减轻,高剂量组治疗效果接近复方磺胺甲恶唑组。研究结果表明,参麦与黄芪注射液对肺孢子虫有明显的治疗作用。