抗病型中草药添加剂对肉仔鸡生产性能及血液指标的影响

本次试验选用1日龄的AA肉仔鸡1000只作为试验动物,将其随机分为3组,每组4个重复,每个重复50只鸡。试验1组和试验2组分别添加0.5%和1.0%的抗病型中草药(鱼腥草、板蓝根、黄柏、白芍等12种中草药)。中草药以粉碎散剂的形式添加到肉仔鸡的日粮中。试验期为6周,每周称重1次。对添加不同类型和不同水平的中草药对肉仔鸡的生产性能、屠宰性能、肉质中药物残留及血液指标的影响进行研究。

籼型抗病恢复系绵恢2009的选育与应用

用IR15 44 明恢 63∥多系 1号杂交选育而成的杂交水稻籼型恢复系绵恢 2 0 0 9,具有恢复力强 ,配合力高 ,米质较优 ,稻瘟病抗性较强 ,制种产量高 ,杂种优势强等特点 ,与野败型、冈型、印尼水田谷型等各类不育系组配 ,均表现出较强的杂种优势 ,所配组合冈优 2 0 0 9、红优 2 0 0 9、C优 2 0 0

免疫型作物抗病剂

该项目是在国家“863”项目、山东省重点攻关项目、山东大学青年基金项目的资助下,依据病原、宿主、环境三要素间相互作用特点,综合运用植物细胞修复、生物膜稳定、抗病诱导、生长发育调控和均衡营养等多种生物技术。

玉米抗腐霉茎腐病种质标记基因型鉴定与遗传多样性分析

腐霉茎腐病(Pythium stalk rot)是玉米生产上的重要病害。本研究利用14个与8个抗玉米茎腐病基因连锁的分子标记对196份抗腐霉茎腐病玉米种质进行抗病标记基因型鉴定,并采用42对多态性SSR标记对54份抗病自交系进行遗传多样性分析,以期阐明玉米抗腐霉茎腐病种质的标记基因型和遗传背景,并为资源的有效利用、新基因的挖掘和杂种优势模式确定提供参考信息。14个与抗病基因连锁的分子标记将196份抗性种质鉴定为128种标记基因型,表明存在多样的抗性基因组合方式。191份种质获得与齐319、X178或1145中一个或多个的相同的扩增,表明97.45%种质可能含有与3个抗玉米茎腐病材料相同的抗病基因;粤61、郑653、赤L136、白53和18--14共5份种质均未扩增出与齐319、X178和1145相同的标记基因型,可能携带其他抗茎腐病基因;遗传背景相近的抗性种质分属不同的标记基因型,表明抗病种质携带的抗病基因可能在育种选择中发生了分离。42对多态性SSR引物在54份抗病材料中共检测出119个等位基因(Na),多态位点百分率(PPB)为99.17%,平均有效等位基因数(Ne)为1.7070,平均Nei's基因多样性(H)为0.3999,平均Shannon's信息指数(I)为0.5844,平均多态信息含量(PIC)为0.5527,变幅为0.2061~0.7844;通过UPGMA聚类分析,54份抗病材料被划分为2个类群,共6个亚群中,分别是旅大红骨亚群、BSSS亚群、塘四平头亚群、PA亚群、PB亚群、Lan亚群,表现出较高的遗传多样性。结果表明,我国6个杂种优势群中均含有较为丰富的抗腐霉茎腐病种质资源,其中PA亚群包含的抗病种质最多。

江苏省水稻品种抗稻瘟病基因型的鉴定与分析

通过离体接种的方法,用已知抗瘟基因型反应的22个稻瘟菌株,对江苏省67个水稻品种进行了抗瘟基因型鉴定和分析。结果表明:67个水稻品种可能携带1个或多个抗瘟基因,其中携带抗瘟基因Piz5、Pizt、Pi1、Pi9、Piz、Pi5、Pi3和Pii的水稻品种对22个稻瘟病菌菌株的抗性频率均大于50%。携带Pita2和Pi9这2个基因的水稻品种特占A和携带Pi9基因的水稻品种辐优136对所有稻瘟病菌菌株表现免疫,Pita2和Pi9这2个基因,尤其是Pi9基因对于水稻抗瘟性极为重要,推测特占A和辐优136是较好的抗瘟育种材料;含有Pia基因的水稻品种,如徐稻41926、徐稻40993等品种抗病性差,极易丧失抗病性。建议生产中推广种植携带Pi9基因的水稻品种,减小携带Pia基因的水稻品种的种植面积。

白车轴草病原菌链格孢菌及其症状研究

通过对不同白车轴草无性系病叶样本的病原菌分离、接种、鉴定及致病力测定,并结合野外观察,研究了安徽铜陵铜尾矿区白车轴草叶斑病病原菌链格孢菌(Alternaria tenuisNees)的致病、发病状况和寄主植物的抗病性、研究表明,不同白车轴草无性系抗病性有明显差异,根据寄主植物发病情况,以及病斑大小、形状、颜色等,将白车轴草无性系分为抗病型(R)和感病型(S)。

美洲黑杨无性系对杨叶锈病的抗性研究

研究了美洲黑杨(Populus deltoides)无性系对杨叶锈病(Melampsora larici-populina)的抗性问题。结果表明,供试的美洲黑杨无性系对杨叶锈病的抗性可分为4种类型,即高抗型、抗病型、感病型和高感型。

Development and Characterization of Monoclonal Antibody Specific to Nuclear Protein of Avian Influenza Virus Type A

Five monoclonal antibodies（Mabs） to nuclear protein of avain influenza virus（AIV） were developed by syncretizing SP 2/0 and the spleen cells from BALB of mice immuized with H9 subtype AIV. Specificity of these Mabs were identified by immunofluorescent assay（IFA） and enzyme linked immunosorbent assay （ELISA）. These five Mabs which were named as AIV-NP-2C3, AIV-NP-6A5, AIV-NP-3 H9, AIV-NP-7B4, AIV-NP-2H4 could react with all viruses of AIV-H9 strains in tests. The result of Western blotting showed that only the 60 ku protein antigen of AIV-H9 could be recognized by the Mabs but never recognized by New castle disease virus, REV and infectious bursa disease virus. The result of preliminary application showed that avian influenza viruses could be deetected bv Mabs in IFA and ELISA. All these Mabs will probably play important roles in preventing and monitoring avian influenza viruses.

Polymorphism Detection of the Twelfth Exon of Equine MxA Gene

[Objective] The study aimed to establish a fast and accurate method to detect the polymorphism of the 12^th exon of equine MxA gene. [Method] The 12^th exon of MxA gene was amplified by mismatch PCR and the products were analyzed by restriction fragment length polymorphism （RFLP） to determine the point mutation at the 1 790 nt of MxA cDNA. The sequence of the PCR products was also analyzed. [Result] There were three genotypes （AA, AB and BB） in the 12^th exon of equine MxA gene; the 2 081 nt of MxA cDNA mutated from G to C, correspondingly changing the 562^th amino acid of the coding region of MxA protein from tryptophan to cysteine; the specific sequence of the PCR products amplified by mismatch PCR-RFLP was consistent with the analysis results of RFLP. [ Conclusion] The mismatch PCR-RFLP was an easy method with accurate results to detect the polymorphism of the 12^th exon of equine MxA gene.

Preparation and Identification of Specific Monoclonal Antibody against Porcine Circovirus Type 2

BALB/c mice were immunized using synthetic tandem polypeptide of Cap protein epitope of porcine circovirus type 2 （PCV2） as the antigen. By using lym-phocyte hybridoma technique, a hybridoma cellline stably secreting monoclonal an-tibody against PCV2-rCap protein was successful y obtained and named as 670#. The ascites titer of the obtained monoclonal antibody was 1∶100 000. Western blot results showed that the monoclonal antibody could react with prokaryotical y ex-pressed PET32a-ORF2 recombinant protein, eukaryotical y expressed ORF1-ORF2 tandem protein and PCV2 whole virus celllysate. Indirect EILSA demonstrated that the monoclonal antibody could bind with ORF1-ORF2 tandem protein. Indirect im-munofluorescence assay （IFA） indicated that the monoclonal antibody could identify native PCV2 virus. The preparation of this monoclonal antibody provided technical tools for epitope analysis and molecular diagnosis of PCV2 virus.

Role of Antivirus Therapy in Treatment of Hepatocellular Carcinoma with Chronic Hepatitis B Infection

Objective： To observe the recurrence and prognosis of hepatocellular carcinoma （HCC） patients coexisting with chronic hepatitis B infection with active virus replication after receiving antivirus therapy using lamivudine and thymosin α1 （Tα1） postoperatively. Methods： From Jan. 2000 to Dec. 2003, 70 patients with HCC coexisting chronic hepatitis B infection with active virus replication were prospectively divided into two groups： control group （n=35） received hepatectomy only; treatment group （n=35） received hepatectomy and lamivudine plus Tα1 therapy postoperatively. The suppression of HBV-DNA, HBeAg seroconverted rate, tumor recurrent rate and the median survival for the two groups were observed and calculated. Results： In treatment group and control group, the 2-year HBV-DNA suppression rate was 100% vs. 4% （P=0.0000）; HBeAg seroconverted rate was 73.0% vs. 7.5% （P〈0.05）; the recurrent rate was 10.0 vs 6.5 months （P=0.0032）; the median survival time was 12.5 vs. 6.0 months （P=0.0023）, respectively. Conclusion： Antivirus therapy using lamivudine and Tα1 postoperatively may suppress the HBV reaction, delay the recurrent time and prolong the survival for HCC patients coexisting chronic HBV infection with active virus replication.

Adiponectin Gene Variation -4522C/T Is Associated with Type 2 Diabetic Obesity and Insulin Resistance in Chinese

The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease （CHD） and type 2 diabetes mellitus （T2DM）. Genotyping of SNP --4522C/T in 304 patients with CHD, 389 patients with T2DM, and 405 age and sex-matched healthy control subjects was carried out by means of PCR-RFLP approach. No significant difference in the genotype or allele frequencies was found, either between patients with CHD and control subjects, or between patients with T2DM and control subjects. However, in the subgroup analysis, an association of the TAr genotype and T allele with type 2 diabetes combined with obesity （BMI ≥ 25 kg/m2） was found （P = 0.014 and P = 0.034, respectively）. Also the homeostasis model assessment of insulin resistance （HOMA-IR） in T2DM patients with T/T genotype was significantly higher than that in T2DM patients carrying C allele （P = 0.0069）. The authors＇ findings for the first time demonstrated that SNP --4522 in the adiponectin gene was associated with T2DM that combined with obesity and higher insulin resistance index in patients with T2DM. This indicated that the variation might associate with an increased susceptibility to type 2 diabetic obesity and insulin resistance. But -4522C/T polymorphism did not contribute to the susceptibility of CHD.

Effect of interferon in combination with ribavirin on the plus and minus strands of HCV RNA in patients with chronic hepatitis C

AIMS To probe the effect of interferon in combination with rib- avirin on the plus and minus strands of hepatitis C virus RNA (HCV RNA). METHODS Twenty-three cases diagnosed as chronic hepatitis C (CHC) according to positive HCV RNA/anti-HCV,fluctuating levels of aminotransferase activities (>1 year) and absence of other hepatitis virus marker,were studied. Among them,13 pa- tients received combined antiviral therapy (subcutaneous injection of 3MU of interferon-α three times per week for 3 months and intra- venous drip of 1 g of ribavirin per day during the first month of treatment with interferon) and 10 patients received single interfer- on therapy (the same as above-mentioned) as control. The plus and minus strands of HCV RNA in sera and peripheral blood mononuclear cells (PBMCs) of these patients were tested by means of nested reverse transcription-polymerase chain reaction (nested RT-PCR). RESULTS At the end of therapy,the abnormal ALT levels de- creased to normal range in 9 (69.23%) cases in the combined antiviral group. Of them,5 (55.56%)experienced post-therapy relapse and 4 (44.44%) were complete responders. In the inter- feron group,the ALT decreased to normal in 6 (60%) cases,of which,4 (66.67%) had post-therapy relapse and 2 (33.33%) were complete responders. The differences between the two groups were nonsignificant (P>0.05). At the end of therapy,the positive rate of the plus strand in sera decreased from 92.3% to 38.46% (P<0.05) and that of the minus strand in PBMCs,from 76.92% to 38.46% (P<0.05) in the combined antiviral group; and in the interferon group,the former decreased from 100% to 50% (P<0.05) and the latter,from 90% to 40% (P<0.05). Again,no significant differences were found between groups (P >0.05). The relapse occurred in patients whose plus strand HCV RNA in PBMCs remained positive before and after treatment. CONCLUSIONS Ribavirin could not enhance the antiviral effect of interferon. The absence of HCV RNA in serum does not mean complete clearance of HCV,and its value for evaluating the an- tiviral effect and prognosis is limited. Therefore,it is essential to measure the plus and minus strands of HCV RNA in sera and PBM- Cs simultaneously.

Setting up the Model of Xeno-graft Verse Host Disease

Objective: To observe human to mouse one way mixed lymphocyte(MLC); And to set up the xeno-grats verse host disease Xeno-graft host disease(XGVHD) model,probing its immunologic mechamism.Methods: Mouse splenic lymphocyte were collected in asepsis and treated by mitomycin as activating cell. Human Peripheral blood lymphocytes (hPBL)were separated and gathered as reacting cell; Mouse splenic lymphocyte and hPBL were mixed to incubate for a week. Destroying recipient (mouse) immune system by total body irradiation (TBI) and intraperitoneal injecting CTX、MTX; Separating and collecting hPBL; Injecting hPBL to mouse by caudal vein. Results; ①HPBL in the experiment groups(mixed mouse lymphocyte) proliferated obviously, the amount of 3H-TdR in corporation increased evidently(P<0.05); The mean percentage of CD 4、CD 8、IgG 、IgM positive cells rose markedly. ②Experiment groups,the hPBL were found in the spleen and kidney tissue, fas protein expressing, we occasionally discovered and apoptosis cells.Conclusion: The human to mouse one-way MLC has obvious lymphocyte proliferation. By these means,we succeed in inducing XGVHD and setting up a XGVHD model.

Survival and prognostic factors in hepatitis B virus-related acute-on-chronic liver failure

AIM:To investigate the survival rates and prognostic factors in patients with hepatitis B virus-related acuteon-chronic liver failure(HBV-ACLF).METHODS:Clinical data in hospitalized patients with HBV-ACLF admitted from 2006 to 2009 were retrospectively analyzed.Their general conditions and survival were analyzed by survival analysis and Cox regression analysis.RESULTS:A total of 190 patients were included in this study.The overall 1-year survival rate was 57.6%.Patients not treated with antiviral drugs had a significantly higher mortality[relative risk(RR)=0.609,P=0.014].The highest risk of death in patients with ACLF was associated with hepatorenal syndrome(HRS)(RR=2.084,P=0.026),while other significant factors were electrolyte disturbances(RR=2.062,P=0.010),and hepatic encephalopathy(HE)(RR=1.879,P<0.001).CONCLUSION:Antiviral therapy has a strong effect on the prognosis of the patients with HBV-ACLF by improving their 1-year survival rate.HRS,electrolyte disturbances,and HE also affect patient survival.

High serum leptin is an independent risk factor for nonresponse patients with low viremia to antiviral treatment in chronic hepatitis C

AIM： To determine whether body weight and/or serum leptin were independent predictors of response to antiviral treatment in patients with chronic hepatitis C. METHODS： A retrospective evaluation was performed in 139 patients with chronic hepatitis C treated with interferon （IFN） from 1996 to 2000. Sustained response was defined as negative by hepatitis C virus （HCV） RNA analysis using PCR and normal transaminase at 24 wk after cessation of IFN therapy. Patients who remained positive for HCV RNA at the end of IFN treatment were defined as resistant to IFN therapy. Sex, age, body mass index （BMI） （≥ 25 vs 〈 25）, complication of diabetes mellitus, serum leptin level （≥ 8.0μg/L vs 〈8.0μg/L）, and the stage of liver fibrosis by needle biopsy （F1/F2 vs F3/F4） were examined. RESULTS： Sustained response was achieved in 33 patients （23.7%）, while others failed to show a response to IFN therapy. Overall, the factors associated with sustained antiviral effects were HCV-RNA load, HCV genotype, serum leptin level, and stage of liver fibrosis evaluated by univariate analysis. BMI was not associated with any therapeutic effect of IFN. Multivariate analysis indicated that HCV-RNA load was a significant risk factor, but among the patients with low viremia （HCV-RNA 〈 100 MU/L）, leptin level was an independent risk factor for IFN resistance. Namely, a high level of serum leptin attenuated the effect of IFN on both male and female patients with low viremia. CONCLUSION： High serum leptin level is a negative predictor of response to antiviral treatment in chronic hepatitis C with low viremia.

Antiviral treatment of hepatitis B virus-transgenic mice by a marine organism, Styela plicata

AIM： To evaluate the antiviral effect of the effective ingredient of Styela plicata in a murine model of hepatitis B virus carrier. METHODS： HBV-transgenic mice were divided into 3 groups （control group, lamivudine treatment group and the effective ingredient of Styela plicata treatment group） and assigned to receive normal diet, lamivudine or the effective ingredient of Styela plicata for consecutive weeks. Serum hepatitis B surface antigen was detected by enzyme-linked immunosorbent assay （ELISA） method. Serum HBV DNA was detected by real-time polymerase chain reaction （RT-PCR）. Serum T helper （h） 1 cytokine interleukin （IL）-2 and Th2 cytokine IL-6 were detected by the quantitative sandwich enzyme immunoassay technique. Another group of HBV-transgenic mice was assigned to receive the effective ingredient of Styela plicata for consecutive weeks. The histology of liver tissue was evaluated before and after treatment. RESULTS： Twelve weeks after starting the therapy, serum hepatitis B surface antigen was significantly lowered in Styela plicata -treated mice and lamivudine-treated mice compared with the mice receiving normal diet （F12wk = 88.81, P12wk = 0.000 〈 0.01）. Serum HBV DNA was significantly lowered in Styela plicata -treated mice and lamivudine-treated mice compared with the mice receiving normal diet （F12wk = 20.71, P12wk = 0.000 〈 0.01）. However, like lamivudine, the effective ingredient of Styela plicata could not inhibit the replication of HBV completely. A rebound phenomenon of hepatitis B surface antigen and HBV DNA in sera could be found 4 wk after withdrawal of medication. Eight weeks after starting the therapy, serum levels before and after Styela plicata treatment of IL-2 were 2.41 ± 0.38 and 10.56 ± 0.78 ng/L, respectively （t8wk = -16.51, P8wk = 0.000 〈 0.01）. Compared with the serum levels of IL-2 in the normal diet-treated mice （2.48 ± 0.17 ng/L; t8wk = 13.23, P8wk = 0.000 〈 0.01）. Serum levels before and after Styela plicata treatment of IL-6 were 63.62 ± 6.31 and 54.52 ± 6.22 ng/L, respectively, compared with the serum levels of IL-6 in the normal diet-treated mice （60.84 ± 4.21 ng/L）. Histological analysis of liver from Styela plicata-treated HBV-transgenic mice also showed catabatic status in inflammation and hepatitis B surface antigen. CONCLUSION： Styela plicata may be an effective anviral medicine in treating chronic hepatitis B.

Steatosis and insulin resistance in hepatitis C: A way out for the virus?

The hepatitis C virus (HCV) induces lipid accumulation in vitro and in vivo. The pathogenesis of steatosis is due to both viral and host factors. Viral steatosis is mostly reported in patients with genotype 3a, whereas meta-bolic steatosis is often associated with genotype 1 and metabolic syndrome. Several molecular mechanisms responsible for steatosis have been associated with the HCV core protein, which is able to induce gene expres-sion and activity of sterol regulatory element binding protein 1 (SREBP1) and peroxisome proliferator-activat-ed receptor γ (PPARγ), increasing the transcription of genes involved in hepatic fatty acid synthesis. Steatosis has been also implicated in viral replication. In infected cells, HCV core protein is targeted to lipid droplets which serve as intracellular storage organelles. These studies have shown that lipid droplets are essential for virus assembly. Thus, HCV promotes steatosis as an eff icient mechanism for stable viral replication. Chronic HCV in-fection can also induce insulin resistance. In patients with HCV, insulin resistance is more strongly associated with viral load than visceral obesity. HCV seems to lead to insulin resistance through interference of intracellular insulin signalling by HCV proteins, mainly, the serine phosphorylation of insulin receptor-1 (IRS-1) and im-pairment of the downstream Akt signalling pathway. The HCV core protein interferes with in vitro insulin signal-ling by genotype-specif ic mechanisms, where the role of suppressor of cytokine signal 7 (SOCS-7) in genotype 3aand mammalian target of rapamycin (mTOR) in geno-type 1 in IRS-1 downregulation play key roles. Steatosis and insulin resistance have been associated with f ibrosis progression and a reduced rate of sustained response to peginterferon plus ribavirin.