乙型肝炎病毒大蛋白检测的临床应用

目的通过检测乙型肝炎患者血清中乙型肝炎病毒外膜大蛋白(HBV-LP)、HBV DNA、乙型肝炎病毒标志物(HBV M)以及乙型肝炎病毒前S1蛋白(PreS1),探讨乙型肝炎病毒外膜大蛋白(HBV-LP)检测对于判断乙型肝炎病毒(HBV)复制的意义。方法采用实时荧光定量PCR法对156例HBV感染者血清中的HBV DNA进行检测,并用ELISA法对HBV-LP、HBV M及PreS1进行检测。结果HBV-LP的检出率与HBV DNA的检出率无明显差异,HBV-LP的含量与HBV DNA拷贝数的对数值具有相关性(γ=0.926,P<0.001),在HBV DNA阳性血清中HBV-LP阳性率(92.4%)明显高于HBeAg(53.8%)和PreS1(56.8%),差异有统计学意义(P﹤0.05)。结论HBV-LP能反映乙型肝炎患者体内HBV复制情况,比PreS1和HBeAg更具有优势,可以作为判断HBV复制新的血清学指标。

提高种蛋分级后大蛋孵化率的研究

为了满足客户需求,保证雏鸡均匀度,我们将种蛋人工分成三个级别,即大蛋(65~70g)、中蛋(57~64g)和小蛋(53~56g),然后根据种蛋的分级进行入孵,以确保雏鸡均匀度在85%以上。通过此种方法保证了雏鸡的均匀度,但在操作过程中发现大蛋(占总种蛋数量的11.2%)的孵化率比其他分级种蛋的孵化率低1%~3%。为此我们设计了试验方案,试验分为两个组,试验组为大蛋专机孵化,对照组为大蛋与小蛋隔批入孵,通过试验组和对照组试验数据对比发现大蛋专机孵化效果比大蛋与小蛋隔批入孵较好,孵化率提高0.6%,健母率提高0.4%。

乙肝病毒表面大蛋白检测在慢性乙型肝炎诊断中的应用价值

目的分析乙肝病毒表面大蛋白（LHBs）和慢性乙型肝炎（CHB）血清标志物之间的关系，探讨LHBs对CHB诊断的临床价值。方法：研究对象124例接受抗病毒治疗的慢性HBV患者，为124例表面抗原（HBsAg）阳性至少6个月的CHB患者，采用酶联免疫吸附试验（ELISA）和聚合酶链反应（PCR）检测CHB血清标志物和LHBs，同时检测HBV—DNA。结果：在慢性非活动性乙型肝炎（1H）诊断中，LHBs和RBV—DNA具有良好的一致性（P〉0．05），而在CAH中，LHBs比HBV—DNA具有更高诊断性，组间比较有统计学差异（P〈O．05）。不同慢性活动性乙型肝炎（CHB）血清标志物中，LHBs的阳性率均比HBV—DNA高，说明LHBs对诊断CHB均有较高的敏感性。HBsAg（＋）HBeAg（＋）HBcAh（＋）、HBsAg（＋）HbeAh（＋）HBcAb（＋）和HBsAg（＋）HBeAg（＋）组间阳性率比较有显著性统计学意义（P〈0．01）。HBV—DNA与LHBs对诊断HBsAg（＋）HBcAh（＋）也具有良好的一致性（P〉0．05）。结论：LHBs可以作为辅助诊断CHB的一项血清学指标。

有效提问 点亮精彩课堂——以《小老鼠运大蛋》语言活动教学为例

我国著名教育家陶行知先生说：＂发明千千万,起点是一问。禽兽不如人,过在不会问。智者问得巧,愚者问得笨。＂诚然,学起于思,思源于疑。提问是一种教育智慧、一种教学艺术,也是教学过程中的一个重要环节,更是提高教学质量的有效手段。教师在设计问题时要做到由易到难,由浅入深,层层推进,从幼儿的认知水平出发,将提问的＂坡度＂转向提问的＂难度＂,这样的课堂提问才能够促进幼儿的发展,使课堂教学精彩纷呈。

奇怪的大蛋

随后的一个礼拜，日子慢得可怕。我估计我差不多每半小时都要出去看一次。在杰默博士说过那蛋可能孵化出东西之后，我迫切地盼望它能孵出个东西来。但是我每一回去看鸡窝，那蛋总是躺在那里，还是一个半月来的老样子。就连那鸡也露出了不耐烦的神气，好像真对那东西能不能孵出什么来并不在乎了。这种迹象太糟糕了，因为结局已在眼前，不该是散伙的时候。如果它现在罢了工，我说不定会自己去孵的。

A Proteinase from Mung Bean Sprouts That Inactivaties Soybean Trypsin Inhibitor

By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h.

Effects of Two Kinds of Heavy Metals Including Cr^(6+) and Pb^(2+) on Garlic Germination

[Objective] This study aimed to revealing the effects of two kinds of heavy metals including Cr6＋ and Pb2＋ on garlic germination.[Method] Root length,enzyme（SOD and CAT）activity and soluble protein content during garlic germination were determined under the stress of two kinds of heavy metals including Cr6＋ and Pb2＋.[Result] During garlic germination,there was an obvious inhibition on buds and seedlings when the concentration of heavy metals including Cr6＋ and Pb2＋ was more than 20 mg/L,while SOD and CAT had certain buffering ability to the heavy metal toxicity with low concentration.With the stress of Cr6＋ in low concentration,SOD activity increased with the increase of metal ion concentration,and then reached maximum when the treatment concentration was 50 mg/L,while CAT activity gradually increased when Cr6＋ concentration was lower than 20 mg/L.With the high concentration of Cr6＋（more than 50 mg/L）or the Pb2＋ stress,SOD and CAT activity continued to decrease with the increase of concentration of heavy metal ions.With various concentrations of heavy metal（Cr6＋ and Pb2＋）solution,soluble protein content showed the decreasing trend with time prolong,and the decreasing trend was obvious with the increase of concentration.Moreover,the tolerance of garlic on Cr6＋ toxicity was much higher than that on Pb2＋.[Conclusion] Two kinds of heavy metals including Cr6＋ and Pb2＋ have impacts on four factors mentioned above during garlic germination,and this study provides a theoretical basis for further discussion of the toxic mechanism of heavy metals on plants.

Characterization of the Full-length cDNA, Chromosomal Localization, and Polymorphism of the Porcine RPLP0 Gene

RPLPO gene encodes the acidic ribosomal phosphoprotein large P0 subunit, which is a component of the 60S subunit. The full-length cDNA sequence of porcine RPLPO was obtained from skeletal muscle of fetal pig cDNA library and deposited in GenBank. The nucleotide sequence and the predicted protein sequence shared high sequence identity with other mammalian homologues. A C/A single nucleotide substitution in exon 5 was detected as Csp6 Ⅰ polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） shows allele frequency diversity among Tongcheng, Xiaomeishan, Yushan, Large White, Landrace, and Duroc breeds. Analyses of somatic cell hybrid panel （SCI-IP） and radiation hybrid （IMpRH） panel showed that the RPLPO gene was mapped to SSC 14q22-q24 and was closely linked to locus SW1321 （25 cR, LOD = 14.54）.

Determination of Isoflavone from Soybean Lines Cultivated in Jilin Province and Correlation Analysis between Isoflavone Content and Soybean Quality

[Objective]The aim of this study was to set up a high performance liquid chromatography for rapid determination of isoflavones from soybean and analyze the correlation between isofalvone content and protein or fat content. [Method]The isoflavones were firstly extracted by 80% methanol and then hydrolyzed at 100 ℃. The chromatographic separation adopted a reversed-phase C18 analytical column with binary high-pressure gradient elution,while its analysis time was 25 min and column temperature was 40 ℃. The diode array detector was used for monitoring with wavelength of 260 nm. The correlation between isofalvone content and protein or fat content was analyzed by data processing system Origin 6.0. [Result]The high performance liquid chromatograph for determination of isoflavones from soybean was verified to be accurate and reliable by methodology. The isoflavones of 85 soybean lines cultivated in Jilin Province were determined,and the results primarily showed the characters and ranges of isoflavones from soybean lines cultivated in Jilin Province,while the isoflavone content of soybeans ranged from 2.29 to 4.89 mg/g,and the average content was 3.36 mg/g. The isoflavone content of 5 soybean lines exceeded 4 mg/g,while there was a remarkably negative correlation between isoflavone content and protein content,and there was no significant positive correlation between isoflavone content and fat content. [Conclusion]The isoflavone content of soybean lines cultivated in Jilin Province is higher,so it is feasible for breeding the soybean lines with high isoflavone content and fat contetnt.

Transient Expression of BYDV-MP in Nicotiana benthamiana

[Objective]The aim of this study was to identify transient expression of movement protein （MP） gene in Nicotinana benthaminana rapidly and further investigate the function of this exogenous gene. [Method]The movement protein gene of barley yellow dwarf virus （BYDV） was cloned into potato virus X （PVX） viral vector of pGR107,and PVX-recombinant vector was obtained. After electroporation of Agrobacterium tumefaciens,PVX was inoculated into the lower leaves of tobacco by Agrobacterium infiltration assay to observe the infection of virus on tobacco. [Result]After infection for 7 days,upper non-inoculated leaves of tobacco infected by the PVX-recombinant vector showed the virus infection symptoms,while the control group had no viral infection phenomenon. Daily follow-up observations for two groups revealed that tobacco infected by PVX-recombinant vector had severe symptoms of virus infection and curling leaves,or even led to necrosis both in infiltrated and systemic leaves in late period. However,tobacco infected by PVX vector had only slight symptoms of virus infection and could recover from infection. RT-PCR of the infected tobacco indicated that exogenous gene BYDV-MP had a normal transcription and expression in tobacco. [Conclusion]As a determinant factor for viral disease,BYDV-MP promotes the systemic infection rate of PVX and its symptom. In addition,it is feasible to express exogenous MP gene in Nicotiana benthaminan via PVX expression vector.

Composition Analysis of HMW-GS in Australian Wheat Cultivars

[Objective] The aim was to provide information for the breeding of wheat in China.[Method] 64 wheat cultivars from Australia were analyzed by SDS-PAGE to determine their high molecular weight（HMW） glutenin subunit combinations.Then,the cultivars had good HMW-GS were screened.[Result] A total of nine alleles were identified at Glu-1.Glu-A1 encoded two subunits,1 and null subunit,and 1 was the major type,the frequency was 59.37%;Glu-B1 encoded five subunits,7,7＋8,7＋9,14＋15,17＋18,and 7＋8 was the major type,the frequency was 56.2%;Glu-D1 encoded two subunits,2＋12,5＋10,and 2＋12 was the major type,the frequency was 70.3%.Furthermore,12 subunit combinations were detected,and the composition of ＂1,2＋12,7＋8＂ was the major type.The quality scores of these cultivars ranged from 4 to 10,with an average of 7.4.[Conclusion] The quality of these varieties was good.

Study on Outer Membrane Protein Patterns of Escherichia coli O38,O53 and O75 Isolated from Chickens

[Objective] This study aimed to investigate the outer membrane protein （OMP） patterns of Escherichia coli 038, 053 and 075 isolates from chickens. [Method] Eight pathogenic E. coil isolates with various serotypes were used as experimental materials to extract OMP by using supersonic schizolysis method and Sarcosyl. After SDS-PAGE electrophoresis, OMP patterns of the extracted products were determined based on the OMP model diagram. [Result] OMP of eight E. coil isolates with three serotypes were divided into three patterns, to be specific, 2 075 isolates respectively belonged to OMP-I and OMP-II pattern, 1 053 isolate belonged to OMP-II pattern, and 5 038 isolates belonged to OMP-I and OMP-III pattern. [Conclusion] Experimental results showed that E. coli isolates with the same serotype may belong to completely different OMP patterns, while serologically unrelated isolates may belong to the same OMP pattern. OMP of E. coil isolates with the same serotype may generate genetic differentiation; in addition, OMP of E. coli isolates with different serotypes may have different genetic correlation.

Characterization of ST13 Protein Expression in Human Colorectal Cancer Tissues

Objective: To characterize the expression of ST13 protein in human tissuesfor investigation of the function of colorectal cancer related gene ST13. Methods: ST13 ORF wascloned and over-expressed in E.coli. The recombinant ST13 protein was purified by affinitychromatography. ST13 monoclonal antibodies were generated and affinity purified with the recombinantprotein. Immunoblot and immunohistochemical staining were employed to analyze ST13 proteinexpression in human tissues. Results: The expression and purification of the recombinant ST13protein were confirmed by SDS-PAGE. The protein yield reached about 2.5 mg/L of induced bacterialculture with a purity of 91.3%. Three strains of hybridoma were obtained with antibody titers from10~4 to 10~5 in ascites fluids and with high specificity for ST13 protein. Immunoblot showed thatthe apparent Mr of ST13 protein in SW480 cells and human tissues estimated by SDS-PAGE mobility wasapproximately 50 000, which was about 10 000 larger than the 41 324 calculated, but theglycosylation of the protein was excluded. Computer modeling revealed the protein to be ahydrophilic molecule. Immunohistochemical staining showed that ST13 protein was evenly distributedin cytoplasm and expressed in colon, stomach, liver, and other epithelial cells. Differences in thestaining intensity of the protein were observed between normal and cancer tissues as well as amongdifferent normal or carcinoma tissues. Conclusion: ST13 protein is a cytoplasmic molecule with anapparent Mr of 50 000. The protein is expressed in colorectal and other epithelial tissues. Theexpression level of the protein is down-regulated in colorectal cancer and varies among differentnormal and/or carcinoma tissues. Comparison of cDNA sequences and protein characteristics indicatesthat ST13 protein and hsp70-interacting protein (Hip) are same proteins, raising the possibilitythat ST13 protein is involved in the development of colorectal cancer through Hsp70 molecularchaperone machinery.

Analysis of Wettability and X-ray Photoelectron Spectroscopy of Wheat Straw/SPI

[Objective] The aim was to improve the adhesive bonding property of wheat straw surface to prepare wheat straw particleboard of soy protein isolate （SPI） adhesive through chemical and enzyme treatments. [Method] Evaluation and analysis were made on wettability of wheat straws in the control group and treated groups （chemical and enzyme treatments） by means of measurement of contact angle and calculation of spreading-penetration parameters （K）. In addition, we made analysis on surface elements through X-ray photoelectron spectroscopy （XPS）. [Result] The re- sults showed that K value of straw treated with sodium hydroxide, hydrogen peroxide and lipase increased by 58.0%, 48.7% and 83.2% compared to that of control group, respectively. The XPS analysis indicated that rapid decrease of silicon content and destruction of wax layer greatly contributed to wettability improvement of wheat straw surface. [Conclusion] The chemical and lipase treatments of wheat straw provided technical support for manufacture of wheat straw particle boand.

Localization of Two GFP_tagged Tobacco Plastid Division Protein NtFtsZs in Escherichia coli

Two plastid division genes, NtFtsZ1 and NtFtsZ2 isolated from Nicotiana tabacum L. were fused with gfp and expressed in Escherichia coli . The regular localizations of full length NtFtsZs∶GFP along the filamentous bacteria indicated that the NtFtsZs could recognize the potential division sites in E. coli and be polymerized with heterogeneous FtsZ from bacteria. The overexpression of NtFtsZs ∶ gfp inhibited the division of host strain cells and resulted in the long filamentous bacterial morphology. These results suggested that eukaryotic ftsZs have similar function to their prokaryotic homologs. Meanwhile, the different deletions of motifs of NtFtsZs are also employed to investigate the functions of these proteins in E. coli . The results showed that the C_terminal domains of NtFtsZs were related to the correct localization of NtFtsZs in E. coli and the N_terminal domains of NtFtsZs were responsible for the polymerization of homogeneous and heterogeneous FtsZ proteins. The significance of these results in understanding the functions of NtFtsZs in plastid division were discussed.

Characterization of a Cyclophilin cDNA from Soybean Cells

A cDNA clone encoded for cyclophilin (GmCyp1) was isolated by RT-PCR method from suspension-cultured soybean (Glycine max L.) cells. The deduced amino acid sequence was 91% identical to a kidney bean cyclophilin in the open reading frame of the gene. Results from Southern blotting analysis suggests that the GmCyp1 belong to a small gene family in soybean cells. The time course of GmCyp1 mRNA accumulation upon treatment of elicitor from yeast extract did not show significant change in the time period examined. The data suggest that the GmCyp1 was not regulated much by biotic factors. A possible role of the cyclophilin in the plant-pathogen interaction was discussed.