钙结合蛋白S100A13在肝癌组织中的表达及其临床意义

目的:探索钙结合蛋白S100A13(S100 calcium binding protein A13,S100A13)在肝癌(hepatocellular carcinoma,HCC)组织中的表达及其临床意义。方法:收集广西医科大学第一附属医院34例HCC及相应癌旁组织、18例正常肝组织标本,商业购置197例肝癌及相应癌旁高密度组织芯片。应用原位RT-PCR技术检测HCC组织、癌旁组织、正常肝组织中S100A13 mRNA的表达,使用免疫组织化学技术检测HCC组织、癌旁组织、正常肝组织中S100A13蛋白的表达,并用Image-ProPlus软件分析免疫组织化学光密度值。结果:S100A13 mRNA在HCC、相应癌旁和正常肝组织中表达的阳性率分别为70.21%、51.06%和33.33%;S100A13 mRNA定位主要在胞核,多表达于核膜。S100A13蛋白在HCC、相应癌旁和正常肝组织中表达的阳性率分别为55.84%、38.96%及26.32%,HCC组织中的平均D值最高(0.038±0.051),其次是癌旁组织(0.022±0.034),正常肝组织(0.01±0.009)最低(P<0.05);S100A13蛋白主要表达于HCC细胞的胞质中,部分细胞核偶见表达,此外在癌旁次生胆小管中及部分炎细胞中也存在高表达。S100A13蛋白在HCC中的表达与患者的性别、年龄及病理分级无关。结论:HCC组织高表达S100A13蛋白,S100A13可能具有作为肝癌治疗靶点的潜力。

Cytomegalovirus frequency in neonatal intrahepatic cholestasis determined by serology, histology, immunohistochemistry and PCR

AIM: To determine cytomegalovirus (CMV) frequency in neonatal intrahepatic cholestasis by serology, histological revision (searching for cytomegalic cells), immunohistochemistry, and polymerase chain reaction (PCR), and to verify the relationships among these methods.METHODS: The study comprised 101 non-consecutive infants submitted for hepatic biopsy between March 1982 and December 2005. Serological results were obtained from the patient's f iles and the other methods were performed on paraffin-embedded liver samples from hepatic biopsies. The following statistical measures were calculated: frequency, sensibility, specif ic positive predictive value, negative predictive value, and accuracy.RESULTS: The frequencies of positive results were as follows: serology, 7/64 (11%); histological revision, 0/84; immunohistochemistry, 1/44 (2%), and PCR, 6/77 (8%). Only one patient had positive immunohistochemical fi ndings and a positive PCR. The following statistical measures were calculated between PCR and serology: sensitivity, 33.3%; specificity, 88.89%; positive predictive value, 28.57%; negative predictive value, 90.91%; and accuracy, 82.35%. CONCLUSION: The frequency of positive CMV varied among the tests. Serology presented the highest positive frequency. When compared to PCR, the sensitivity and positive predictive value of serology were low.

SPLUNC1基因在2种绵羊不同组织器官中差异表达分析

SPLUNC1是抗病原微生物感染的关键蛋白。为研究SPLUNC1基因在2种绵羊不同组织器官中的差异表达情况,试验选取健康新疆阿勒泰羊5只,多胎萨福克羊5只,放血处死后分别采集气管、支气管、肺脏、胃、食管、结肠、心脏、脾脏、皮肤,提取RNA,反转录合成cDNA。以cDNA为模板,扩增SPLUNC1基因748 bp特征片段,克隆至pMD19-T载体中,提取质粒。以梯度稀释的质粒为模板,建立SPLUNC1基因转录本拷贝数的绝对定量标准曲线,对2种绵羊各组织器官中SPLUNC1的mRNA初始拷贝数进行定量分析。结果表明:阿勒泰羊标准曲线扩增效率(E)为97%,回归系数(R^(2))为0.9990,SPLUNC1 mRNA初始拷贝数从高至低依次为气管、支气管、肺脏、脾脏、食管、心脏、胃、结肠、皮肤。萨福克羊标准曲线扩增效率(E)为99%,回归系数(R^(2))为0.9989,SPLUNC1 mRNA初始拷贝数从高至低依次为气管、支气管、肺脏、脾脏、心脏、食管、胃、结肠、皮肤。2种绵羊的气管SPLUNC1 mRNA表达量均显著高于其他各组织(P<0.05),皮肤表达量最低。新疆阿勒泰羊的9种组织器官该基因表达量均显著高于多胎萨福克羊(P<0.05)。上述结果说明绵羊的SPLUNC1基因的分布具有组织器官特异性,为揭示绵羊抗感染的分子调控机制提供一定参考。

Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata

Accurate quantification of transcripts using quantitative real-time polymerase chain reaction （qPCR） depends on the identification of reliable reference genes for normalization. This study aimed to identify and validate seven reference genes, including actin-2 （ACT-2）, elongation factor 1 alpha （EF-1α）, elongation factor 1 beta （EF-1β）, glyceraldehyde-3-phosphate dehydrogenase （GAPDH）, ubiquitin （UBQ）, β-tubulin （β-TUB）, and 18 S ribosomal RNA, from Crassostrea angulata, a valuable marine bivalve cultured worldwide. Transcript levels of the candidate reference genes were examined using qPCR analysis and showed differential expression patterns in the mantle, gill, adductor muscle, labial palp, visceral mass, hemolymph and gonad tissues. Quantitative data were analyzed using the geNorm software to assess the expression stability of the candidate reference genes, revealing that β-TUB and UBQ were the most stable genes. The commonly used GAPDH and 18S rRNA showed low stability, making them unsuitable candidates in this system. The expression pattern of the G protein β-subunit gene （Gβ） across tissue types was also examined and normalized to the expression of each or both of UBQ andβ-TUB as internal controls. This revealed consistent trends with all three normalization approaches, thus validating the reliability of UBQ and β-TUB as optimal internal controls. The study provides the first validated reference genes for accurate data normalization in transcript profiling in Crassostrea angulata, which will be indispensable for further fimetional genomics studies in this economically valuable marine bivalve.

肺曲霉球1例及实验研究

目的对1例未进行真菌培养的肺真菌球患者术后的组织病理进行实验研究以确定病原菌属。方法 1例咯血并左肺上叶占位的患者接受了左肺上叶切除术,术后组织病理诊断肺真菌球,曲霉感染可能性大,但病变组织未进行真菌培养。本研究对其组织病理标本分别进行了真菌荧光染色、免疫组化染色和组织PCR。结果真菌荧光染色清楚地显示亮蓝色有隔菌丝,呈锐角分支;免疫组织化学染色提示病原菌为曲霉属;组织PCR检测结果阴性。结论当病变组织未进行真菌培养时,免疫组化技术有助于确定病原菌属。

小叶杨GA20氧化酶基因的克隆、表达及单核苷酸多态性分析

利用RT-PCR方法,首次从小叶杨未成熟木质部cDNA中分离出PsGA20OxcDNA全长及其基因组DNA,并进行测序和序列分析。结果表明:克隆的小叶杨PsGA20OxcDNA片段总长为1403bp,基因内部含有完整的开放阅读框架,大小为1155bp,编码区可编码长度为385个AA残基,所推导的蛋白质氨基酸序列与拟南芥和水稻GA20Ox蛋白的同源性分别为66.0%和57.0%。组织特异性RT-PCR结果显示,PsGA20Ox基因在杨树茎、叶片和顶端分生组织中均有表达,但其表达模式却不同:PsGA20Ox在成熟木质部中表达丰度最高,在未成熟木质部和嫩叶中表达丰度较高,在顶端分生组织中有少量表达,在形成层组织中表达丰度极低。在此基础上,组合利用MEGA4.0和DnaSP4.50.4软件对小叶杨36株基因型个体的PsGA20Ox基因组DNA序列进行比对和分析,检测到49个单核苷酸多态性(SNP)位点,频率为1/35bp。其中,15个是常见SNPs,34个为罕见SNPs。在这些SNPs中,37个属于转换,12个属于颠换。在外显子区域,共检测到26个SNP位点,其中23个为同义突变,3个为错义突变。对PsGA20Ox基因内SNPs进行的连锁不平衡分析表明,随着核苷酸序列长度的增加,SNPs的连锁不平衡在基因内部就迅速衰退,因此,在小叶杨中,基于候选基因的连锁不平衡作图是可行的,而基于整个杨树基因组的连锁不平衡作图是不可行的,也是不必要的。

趋化因子CXC受体3在肝细胞癌中的表达及意义

背景与目的:最近研究揭示趋化因子及其受体网络在肿瘤侵袭转移中起重要作用。本研究旨在探讨趋化因子CXC受体3(CXCR3)在肝细胞癌(hepatocellularcarcinoma,HCC)中的表达,及其与HCC临床病理特征的关系。方法:选取7株人HCC细胞系、18例正常肝组织、64例HCC患者的癌组织和癌旁肝组织作为研究对象。采用RT-PCR和实时定量PCR方法,研究这些组织中CXCR3mRNA的表达情况。采用免疫组织化学方法,研究CXCR3蛋白的表达情况和HCC复发转移的关系。结果:在高转移细胞系MHCC97-H中,CXCR3与!2微球蛋白的mRNA拷贝均数之比为33.0×10-4,在低转移细胞系MHCC97-L中,该数值为8.7×10-4,在无转移能力的5株细胞系则未检出CXCR3的mRNA表达。在MHCC97-H和MHCC97-L中均见CXCR3蛋白强阳性染色,而在无转移能力的细胞系中只有HepG2见到微弱染色,其余4株均未见染色。CXCR3蛋白在HCC组织中的强阳性染色,与HCC的侵袭性正相关(P=0.003)。HCC组织中CXCR3蛋白强阳性的患者和CXCR3阴性或弱阳性的患者的1、2年无瘤生存率分别为66.7%、31.3%和75.0%、59.5%,差异有显著性(P=0.044)。结论:CXCR3的表达与HCC的侵袭和转移相关。

食管鳞癌及癌旁粘膜中核转录因子κBp65及IκBα基因和蛋白表达及意义

目的:探讨核转录因子κB(NF-κB)p65基因及其抑制基因IκBα的表达与食管鳞癌发生的关系。方法:用RT-PCR及免疫组化SP法检测了61例食管鳞癌及其正常粘膜上皮、癌旁粘膜上皮中NF-κBp65基因、IκBα基因mRNA及蛋白的表达。结果:食管鳞癌中p65mRNA、IκBαmRNA的表达量明显高于正常上皮(P<0.01),p65蛋白、IκBα蛋白胞浆表达或胞核表达率均明显高于正常上皮(P<0.01)。癌旁粘膜中,NF-κBp65蛋白、IκBα蛋白在癌旁正常上皮中的胞浆或胞核阳性率均明显低于不典型增生上皮、原位癌和浸润癌(P均<0.05)。结论:NF-κBp65、IκBαmRNA表达和蛋白表达均与食管鳞癌的发生有关。

中国麻鸡中发现禽J亚群白血病

首次报道了中国特有鸡种——麻鸡J亚群白血病的发病情况。山东某种鸡场饲养的中国麻鸡,于开产前出现消瘦、贫血、瘫痪等症状,死亡率达10%。经大体剖检发现,病鸡的内脏器官均弥漫性肿大,色彩斑驳,质度较硬;在胸骨内侧、小肠浆膜面和气管粘膜面出现大小不等的肿瘤结节,呈灰白色。组织学检查发现,增生的肿瘤细胞为均一的髓细胞。用禽白血病病毒J亚群(ALV-J)的特异性引物进行PCR检测,阳性率为89%(1517);PCR产物测序,其基因序列、预期氨基酸序列与ALV-J原型株HPRS-103的同源性分别为98.05%和97.4%。用ALV-J单克隆抗体,经免疫组织化学检测发现,在肿瘤组织、肝、脾、肾、骨髓、腺胃中呈现强特异性染色。上述检测表明此髓细胞肿瘤是由ALV-J感染引起的。ALV-J麻鸡病例的发现警示应注意中国地方种鸡的白血病净化工作。

Detection of the SYT-SSX Fusion Gene in Synovial Sarcoma

Objective: To investigate the feasibility and significance of detecting SYT-SSX fusion gene in paraffin-embedded tissues of synovial sarcoma (SS) by reverse-transcriptase polymerase chain reaction(RT-PCR) methods. Methods: Twenty cases of SS tumors from archival materials were collected and all samples were formalin-fixed and paraffin-embedded (FFPE). SYT-SSX fusion transcript was detected by RT-PCR. Home-keeping gene Porphobilinogen Deaminase (PBGD) was regarded as internal control.Results: PBGD mRNA was detected in all 20 tumor cases (100%). SYT-SSX fusion transcript was detected in 18 tumor cases (90%). In 18 SYT-SSX positive SS cases, there are 12 present SYT-SSX1 fusion transcript and 6 present SYT-SSX2 fusion transcript. SYT-SSX1 fusion transcript can be seen in 9 monophasic SS and 3 biphasic SS. In 6 SYT-SSX2 positive SS cases, 4 were monophasic SS and 2 were biphasic. Conclusion: Detection of SYT-SSX fusion transcripts in FFPE tissues for diagnosis of SS is feasible and sensitive. Subtypes of SYT-SSX fusion gene may provide prognosis information.

TACC3 mRNA及蛋白在乳腺癌中的表达及其临床意义

目的:探讨转录相关酸性卷曲蛋白3(transforming acidic coiled-coil proteins,TACC3)mRNA和蛋白在乳腺癌中的表达及其临床意义。方法:收集手术切除乳腺癌及癌旁正常组织标本各85例,进行免疫组织化学染色、RT-PCR、Western blot的检测。结果:乳腺癌组织中的TACC3 mRNA和蛋白的表达水平高于癌旁组织(P<0.05),且TACC3的表达与乳腺癌的分化程度、肿瘤直径、淋巴结的转移、P53表达相关,而与患者的年龄、是否绝经、ER、PR、Ki-67、HER-2等无明显相关。结论:TACC3在乳腺癌中的表达程度与临床病理特征有关,在乳腺癌发生发展过程中可能具有一定的作用。

5-脂氧合酶在小鼠恶唑酮溃疡性结肠炎中的表达研究

目的研究5-脂氧合酶（5-LOX）在恶唑酮诱导溃疡性结肠炎（UC）小鼠结肠黏膜中的表达情况，探讨其在UC发病机制中的作用及其与炎症程度的关系。方法50只Balb／c小鼠分为正常对照组10只和模型组40只。模型组用3％恶唑酮致敏2d，第5天用0．5％恶唑酮灌肠。第8天用颈椎脱臼法处死所有小鼠。实验期间观察小鼠疾病活动指数（DAI），处死小鼠后行结肠组织学病理评分（HPS）。RT—PCR法检测结肠黏膜5-LOXmRNA的表达，免疫组织化学方法检测结肠黏膜5-LOX的蛋白表达情况。结果模型组DAI和HPS均高于对照组，5-LOXmRNA和蛋白表达也明显高于对照组（均P〈001），并随炎症程度加重。表达水平明显增高。结论5-LOX在UC中表达水平明显升高，并参与了UC的发生和发展，在一定程度上可反应疾病的活动度，并且可能作为药物治疗的潜在靶点。

Integration of human papillomavirus 18 DNA in esophageal carcinoma 109 cells

AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.

Study on the Correlation between the Expression of Angiogenic Factor VEGF and Microsatellite Instability in Gastric Adenocarcinoma

Objective: To investigate the correlation between the microsatellite instability (MSI) and the expression of vascular endothelial growth factor (VEGF) in gastric adenocarcinoma. Methods: PCR SSCP method was used to detect MSI of thirty cases with gastric adenocarcinoma at five loci in each patient. Expression of VEGF was examined by the method of immunohistochemistry. Results: The positive of MSI was in 13 patients out of 30 patients (43.4%) in our study. Positive VEGF Immunostaining was detected in 18 patients (60.0%). VEGF was decreased in microsatellite instability high (MSI H) gastric adenocarcinoma. Conclusion: MSI H and microsatellite stable (MSS) gastric adenocarcinoma may follow a different pathway of angiogenesis. The low frequency of VEGF expression among MSI H cancer may partially explain why these cancers are less aggressive.

Identification and characterization of microbial agents in a bloom of floating microorganisms in the Hailang River,China in 2006

In February 2006,a large amount of unknown floating microorganisms appeared in the Hailang River(HR) in the City of Hailin,Heilongjiang Province,China.The microorganisms caused the river water fouling and threatened the clean water supply of the city.To identify the unknown floating organisms,morphological and histological inspection,PCR,cloning and sequence analysis were conducted.The results revealed that the unknown floating organisms in the river were a novel species that likely belonged to oomycetes in Saprolegniales.The organisms were named Saprolegnia sp.HL0602.

Diagnosis of intestinal tuberculosis using a monoclonal antibody to Mycobacterium tuberculosis

AIM:To investigate the utility of immunohistochemical(IHC) staining with an antibody to Mycobacterium tuberculosis(M.tuberculosis) for the diagnosis of intestinal tuberculosis(TB).METHODS:We retrospectively identified 10 patients(4 males and 6 females;mean age = 65.1 ± 13.6 years) with intestinal TB.Clinical characteristics,including age,gender,underlying disease,and symptoms were obtained.Chest radiograph and laboratory tests,including sputum Ziehl-Neelsen(ZN) staining,M.tuberculosis culture,and sputum polymerase chain reaction(PCR) for tubercle bacilli DNA,as well as Tuberculin skin test(TST) and QuantiFERON-TB gold test(QFT),were examined.Colonoscopic records recorded on the basis of Sato's classification were also reviewed,in addition to data from intestinal biopsies examined for histopathological findings,including hematoxylin and eosin staining,and ZN staining,as well as M.tuberculosis culture,and PCR for tubercle bacilli DNA.For the present study,archived formalin-fixed paraffin-embedded(FFPE) intestinal tissue samples were immunohistochemically stained using a commercially available species-specific monoclonal antibody to the 38-kDa antigen of the M.tuberculosis complex.These sections were also stained with the pan-macrophage marker CD68 antibody.RESULTS:From the clinical data,we found that no patients were immunocompromised,and that the main symptoms were diarrhea and weight loss.Three patients displayed active pulmonary TB,six patients(60%) had a positive TST,and 4 patients(40%) had a positive QFT.Colonoscopic findings revealed that all patients had type 1 findings(linear ulcers in a circumferential arrangement or linear ulcers arranged circumferentially with mucosa showing multiple nodules),all of which were located in the right hemicolon and/or terminal ileum.Seven patients(70%) had concomitant healed lesions in the ileocecal area.No acid-fast bacilli were detected with ZN staining of the intestinal tissue samples,and both M.tuberculosis culture and PCR for tubercle bacilli DNA were negative in all samples.The histopathological data revealed that tuberculous granulomas were present in 4 cases(40%).IHC staining in archived FFPE samples with anti-M.tuberculosis monoclonal antibody revealed positive findings in 4 patients(40%);the same patients in which granulomas were detected by hematoxylin and eosin staining.M.tuberculosis antigens were found to be mostly intracellular,granular in pattern,and primarily located in the CD68 + macrophages of the granulomas.CONCLUSION:IHC staining with a monoclonal antibody to M.tuberculosis may be an efficient and simple diagnostic tool in addition to classic examination methods for the diagnosis of intestinal TB.

Expression of ABCG2 in human gastric carcinoma

Objective:The aim of this study was to investigate the expression of ABCG2 in human gastric carcinoma and its clinical significance.Methods:Expression of ABCG2 was examined with immunohistochemical technique in the specimens from 45 gastric carcinoma tissues and 30 surrounding normal tissues.The mRNA expression of ABCG2 was measured by RT-PCR and real-time quantitative PCR in 30 cases of gastric carcinoma and normal gastric mucosa, respectively.Results:ABCG2 expression was observed in 28 of 45(62.2%) cases by immunohistochemical analysis.In ABCG2-positive tumors, adjacent non-neoplastic tissue was similarly analyzed, revealed that ABCG2 was up-regulated in gastric carcinoma.ABCG2 expression in poorly differentiated/undifferentiated carcinoma was significantly higher than that in well/moderately-differentiated carcinoma(P < 0.05).The mRNA expression of ABCG2 was significantly higher than that in normal gastric mucosa(P < 0.05).Conclusion:ABCG2 plays an important role in the multi-drug resistance of gastric carcinoma.ABCG2 might be an important factor in the research of gastric cancer stem cell.

Cloning,tissue distribution and effects of fasting on pituitary adenylate cyclase-activating polypeptide in largemouth bass

Pituitary adenylate cyclase activating polypeptide （PACAP） has a wide range of biological functions. We cloned the full-length cDNAs encoding PACAP and PACAP-related peptide （PRP） from the brain of largemouth bass （Micropterus salmoides） and used real-time quantitative PCR to detect PRP- PACAP mRNA expression. The PRP-PACAP cDNA has two variants expressed via alternative splicing： a long form, which encodes both PRP and PACAP, and a short form, which encodes only PACAR Sequence analysis results are consistent with a higher conservation of PACAP than PRP peptide sequences. The expression of PACAP-Iong and PACAP-short transcripts was highest in the forebrain, followed by the medulla, midbrain, pituitary, stomach, cerebellum, intestine, and kidney; however, these transcripts were either absent or were weakly expressed in the muscle, spleen, gill, heart, fatty tissue, and liver. The level of PACAP-short transcript expression was significantly higher than expression of the long transcript in the forebrain, cerebella, pituitary and intestine, but lower than that of the long transcript in the stomach. PA CAP- long and PACAP-short transcripts were first detected at the blastula stage of embryogenesis, and the level of expression increased markedly between the muscular contraction stage and 3 d post hatch （dph）. The expression of PACAP-long and PACAP-short transcripts decreased significantly in the brain following 4 d fasting compared with the control diet group. The down-regulation effect was enhanced as fasting continued. Conversely, expression levels increased significantly after 3 d of re-feeding. Our results suggest that PRP- PA CAP acts as an important factor in appetite regulation in largemouth bass.

Omentum facilitates liver regeneration

AIM:To investigate the mechanism of liver regeneration induced by fusing the omentum to a small traumatic injury created in the liver. We studied three groups of rats. In one group the rats were omentectomized; in another group the omentum was left in situ and was not activated,and in the third group the omentum was activated by polydextran particles. METHODS:We pre-activated the omentum by injecting polydextran particles and then made a small wedge wound in the rat liver to allow the omentum to fuse to the wound. We monitored the regeneration of the liver by determining the ratio of liver weight/body weight,by histological evaluation (including immune staining for cytokeratin-19,an oval cell marker),and by testing for developmental gene activation using reverse transcription polymerase chain reaction (RT-PCR). RESULTS:There was no liver regeneration in the omentectomized rats,nor was there significant regeneration when the omentum was not activated,even though in this instance the omentum had fusedwith the liver. In contrast,the liver in the rats with the activated omentum expanded to a size 50% greater than the original,and there was histologically an interlying tissue between the wounded liver and the activated omentum in which bile ducts,containing cytokeratin-19 positive oval cells,extended from the wound edge. In this interlying tissue,oval cells were abundant and appeared to proliferate to form new liver tissue. In rats pre-treated with drugs that inhibited hepatocyte growth,liver proliferation was ongoing,indicating that regeneration of the liver was the result of oval cell expansion. CONCLUSION:Activated omentum facilitates liver regeneration following injury by a mechanism that depends largely on oval cell proliferation.

Transgenic Date Palm Containing Endotoxin Cry3Aa Gene

Date palm, like all other crops, is very sensitive to the injury by many insect pests, which may lead to the death of the affected plant and causes decrease in yield. In the present study, an efficient Agrobacterium for genetic transformation was successfully achieved for well known date palm （Phoenix dactylifera L.） cv. Medjool and Khalas using callus as explant. Embryogenic callus were recorded 100% mortality when cultured on MS medium containing 100 mg/L kanamycin with different cultivars, thus it was chosen for the selection of transformed explants. Embryogenic callus of Medjool and Khalas were incubated with Agrobacterium tumefaciens strain LBA 4404 for 0.5, 1, 2, 4 and 24 h on LB medium. After the incubation periods, embryogenic callus was transferred to MS medium with 0.1 mg/L NAA, 0.05 mg/L BA, 250 mg/L carbenicillin and 100 mg/L kanamycin for detection of transgenic embryogenic callus. Polymerase chain reaction （PCR） was used for the rapid screening of Cry3Aa gene. For screening, total genomic DNA was isolated from transformants. Using primer specific to Cry3Aa gene （forward and reverse）, a PCR product with a size of about 2,000 bp was amplified when all nucleic acid from the transformants were utilized as templates. PCR analysis confirmed the appearance of the transgene of 2,000 bp in one individual plantlet. Presence and integration of foreign Cry3Aa gene in regenerated kanamycin resistant embryogenic callus was also confirmed by Southern blot hybridization. It was found that one transgenic embryogenic callus for both Medjool and Khalas showed a single copy of gene integration. These results signify the successful transfer of Cry3Aa gene into date palm plant.