非小细胞肺癌中Reprimo以及14-3-3 Σ基因启动子异常甲基化检测

目的探讨Reprimo以及14-3-3 Σ基因启动子在非小细胞肺癌(NSCLC)中的异常甲基化状态及其与临床病理资料的联系。方法采用甲基化特异性聚合酶链反应(Methylation Specific-PCR,MSP)技术检测60例NSCLC以及癌旁正常组织中Reprimo以及14-3-3 Σ基因启动子异常甲基化状态。结果Reprimo以及14-3-3 Σ基因启动子异常甲基化在NSCLC组织中发生频率分别为36.67%(22/60)和28.33%(17/60),与癌旁正常组织具有显著性差异(P值分别为0.000和0.002)。Reprimo基因启动子异常甲基化频率与吸烟习惯以及年龄相关;14-3-3 Σ启动子异常甲基化与年龄、吸烟习惯、性别、病理类型和临床分期以及淋巴结转移等无相关。结论NSCLC中存在Reprimo和14-3-3 Σ等基因启动子较高频率的异常甲基化。

Molecular Characterization of Cotton 14-3-3L Gene Preferentially Expressed During Fiber Elongation

The 14-3-3 protein, highly conserved in all eukaryotic cells, is an important regulatory protein. It plays an important role in the growth, amplification, apoptosis, signal transduction, and other crucial life activities of cells. A eDNA encoding a putative 14-3-3 protein was isolated from cotton fiber eDNA library. The eDNA, designated as Gh14-3-3L （Gossypium hirsutum 14-3-3-like）, is 1,029 bp in length （including a 762 bp long open reading frame and 5＇-/3＇-untranslated regions） and deduced a protein with 253 amino acids. The GhI4-3-3L shares higher homology with the known plant 14-3-3 proteins, and possesses the basic structure of 14-3-3 proteins： one dimeric domain, one phosphoralated-serine rich motif, four CC domains, and one EF Hand motif. Northern blotting analysis showed that Gh14-3-3L was predominantly expressed during early fiber development, and reached to the peak of expression in 10 days post anthers （DPA） fiber cells, suggesting that the gene may be involved in regulating fiber elongation. The gene is also expressed at higher level in both ovule and petal, but displays lower or undeteetable level of activity in other tissues of cotton.

Expression of the Glypican-3 Gene in α-fetoprotein-negative Human Hepatocellular Carcinoma

Objective： To investigate the expression of Glypican-3 gene in （α-fetoprotein （AFP）-negative hepatocellular carcinoma （HCC） and clarify whether Glypican-3 expression is a useful parameter for the diagnosis of hepatocelhllar carcinoma （HCC）, especially for AFP-negative ones. Methods： Forty-one specimens of AFP-negative hepatocellular carcinoma and para-carcimoma tissue were studied for the expression of Glypican-3 by Northern blot. The expression of Glypican-3 protein was detected immunohistochemically with specific polyclonal antibody. Results： Northern blot analysis indicated that the expression of Glypican-3 mRNA was intensively detected in 30 of 41 AFP-negative HCC （73.17%）. In contrast, Glypican-3 mRNA was only weakly detected in 4 of the surrounding non-tumor liver tissues. There was significant difference in the Glypican-3 mRNA expression in large tumors （〉5 cm） （79.31%） and in small tumors （〈5 cm） （41.67%） （P〈0.01）. Gypican-3 mRNA was more frequently overexpressed in poorly differentiated HCC than in well differentiated ones （76.47% vs. 42.86%, P〈0.05）. The Glypican-3 expression was not correlated with age. sex. ttbsAg seropositivity, fibroeapsule, portal venous embolus and intrahepatic metastasis. The overexpression of Glypican-3 protein in HCC was targeted in tumor cells, not in bile duct cells and other interstitial cells. Conclusion： Glypican-3 was specially overexpressed in AFP-negative HCC, and its expression was closely correlated with the tumor size and tumor grade. Glypican-3 gene may play important roles in hepatocareinogenesis, and can be used as a new biochemical marker of HCC, especially for AFP-negative HCC.

Identification of a Novel VEGFR-3 Missense Mutation in a Chinese Family with Hereditary Lymphedema Type I

A novel mutation of vascular endothelial growth factor receptor gene （VEGFR-3）, was identified in a four-generation Chinese family with hereditary lymphedema type I （HL-I）. Genetic linkage analysis was performed on the known genetic locus for HL-I with a panel of polymorphic markers, and then mutations were screened out by direct sequencing. By genotyping, the family showed the linkage to HL-I locus on 5q35.3. Mutation screening analysis of the exons encoding the intracellular kinase domains of VEGFR-3, revealed a novel missense mutation D1055V. This mutation cosegregated with the disease phenotype in the family and was not found in 100 normal controls. This finding has expanded the spectrum of the VEGFR-3 gene mutations causing HL-I, and will be useful for further genetic consultation and genetic diagnosis.

Ex vivo non-viral vector-mediated neurotrophin-3 gene transfer to olfactory ensheathing glia: effects on axonal regeneration and functional recovery after implantation in rats with spinal cord injury

Objective Combine olfactory ensheathing glia （OEG） implantation with ex vivo non-viral vector-based neurotrophin- 3 （NT-3） gene therapy in attempting to enhance regeneration after thoracic spinal cord injury （SCI）. Methods Primary OEG were transfected with cationic liposome-mediated recombinant plasmid pcDNA3.1 （＋）-NT3 and subsequently implanted into adult Wistar rats directly after the thoracic spinal cord （T9） contusion by the New York University impactor. The animals in 3 different groups received 4x 1050EG transfected with pcDNA3.1 （＋）-NT3 or pcDNA3.1 （＋） plasmids, or the OEGs without any plasmid transfection, respectively; the fourth group was untreated group, in which no OEG was implanted. Results NT-3 production was seen increased both ex vivo and in vivo in pcDNA3.1 （＋）-NT3 transfected OEGs. Three months after implantation of NT-3-transfected OEGs, behavioral analysis revealed that the hindlimb function of SCI rats was improved. All spinal cords were filled with regenerated neurofilament-positive axons. Retrograde tracing revealed enhanced regenerative axonal sprouting. Conclusion Non-viral vector-mediated genetic engineering of OEG was safe and more effective in producing NT- 3 and promoting axonal outgrowth followed by enhancing SCI recovery in rats.

Over-expression of fibroblast growth factor receptor 3 in human hepatocellular carcinoma

AIM: To describe the significant over-expression of fibroblast growth factor receptor 3 (FGFR3), which is a signal transduction and cell proliferation related gene in hepatocellular carcinoma (HCC).METHODS: Following DNA microarray, Northern blot and quantitative real-time PCR were employed to confirm FGFR3 expression difference in HCC tissues and surrounding non-neoplastic liver tissue. FGFR3 expression levels were further determined by immunohistochemical study in 43 cases of HCC.RESULTS: Northern blot results showed the significant over-expression of FGFR3 in HCC tissues, which was consistent with that from DNA microarray. Quantitative real-time PCR demonstrated that the mean ratio of FGFR3 mRNA to glyceraldehyde-3-phosphate dehydrogenase (GADPH) mRNA in HCC tissue was 0.250, whereas the ratio in non-neoplastic liver tissue was 0.014. Statistical analyses of 43 cases of HCC revealed that HCC scored higher than the matched non-neoplastic liver tissues.Examination of clinicopathological features revealed a strong correlation of over-expression of FGFR3 with poor tumor differentiation and high nuclear grade.CONCLUSION: Over-expression of FGFR3 may play an important role in liver carcinogenesis. FGFR3 may be an ideal candidate as a molecular marker in the diagnosis of HCC and a potential therapeutic target.

Expression Analysis of 14-3-3 Gene in Tall Fescue under Several Abiotic Stresses

To study the functions of 14-3-3 gene family in tall fescue, the potential functions of 13 14-3-3 proteins in Arabidopsis were investigated by bioinformatic analysis. Based on the sequences of 14-3-3 genes in tall fescue by transcriptome and proteomic sequencing, the full-length cDNA sequences of 4 14-3-3 genes in tall fescue were obtained. Their sequences were aligned by Clustal W2. The results showed that the genetic relationships between 14-3-3A and 14-3-3D, 14-3-3B and 14-3-3C are closer, and their main structures are very conservative. The changes in expression levels of 14-3-3 genes under low nitrogen, drought, high temperature and high salt stresses were investigated by fluorescence quantitative PCR. The expres- sion level of 14-3-3A makes responses to low nitrogen, drought, high temperature and high salt stresses; the expression levels of other genes also make responses to abiotic stresses in varying degrees, but the relevant response mechanisms are not exactly the same. Therefore, it is speculated that the 14-3-3 gene family regu- lates stress resistance of plants through different pathways, and functional differenti- ation occurs during its evolution.

GATA-3 promotes Th2 responses through three different mechanisms： induction of Th2 cytokine production, selective growth of Th2 cells and inhibition of Thl cell-specific factors

Naive CD4 T cells can differentiate into at least two different types ofT helpers, Thl and Th2 cells. Th2 cells, capable of producing IL-4, IL-5 and IL-13, are involved in humoral immunity against extracellular pathogens and in the induction of asthma and other allergic diseases. In this review, we summarize recent reports regarding the transcription factors involved in Th2 differentiation and cell expansion, including StatS, Gfi- 1 and GATA-3. Stats activation is necessary and sufficient for IL-2-mediated function in Th2 differentiation. Enhanced Stats signaling induces Th2 differentiation independent of IL-4 signaling; although it does not up-regulate GATA-3 expression, it does require the presence of GATA-3 for its action. Gfi-1, induced by IL-4, promotes the expansion of GATA-3-expressing cells. Analysis of conditional Gata3 knockout mice confirmed the critical role of GATA-3 in Th2 cell differentiation （both IL-4 dependent and IL-4 independent） and in Th2 cell proliferation and also showed the importance of basal GATA-3 expression in inhibiting Thl differentiation.

Regulated expression of TATA-binding protein-related factor 3(TRF3)during early embryogenesis

RNA polymerase （Pol） Ⅱ transcription persists in TATA-box-binding protein （TBP）^-/- mutant mouse embryos, indicating TBP-independent mechanisms for Pol Ⅱ transcription in early development. TBP-related factor 3 （TRF3） has been proposed to substitute for TBP in TBP^-/- mouse embryos. We examined the expression of TRF3 in maturing oocytes and early embryos and found that TRF3 was co-expressed with TBP in the meiotic oocytes and early embryos from the late one-cell stage onward. The amounts of TBP and TRF3 changed dynamically and correlated well with transcriptional activity. Chromatin immunoprecipitation （CHIP） assay revealed that different gene promoters in mouse embryonic stem （ES） cells recruited TRF3 and TBP selectively. Comparative analyses of TRF3 and TBP during cell cycle showed that both factors proceeded through cell cycle in a similar pace, except that TRF3 was slightly delayed than TBP in entering the nucleus when cells were exiting the M-phase. Data from expression and biochemical analyses therefore support the hypothesis that TRF3 plays a role in early mouse development. In addition, results from co-localization study suggest that TRF3 may be also involved in Pol Ⅰ transcription.

Increased DNA binding activity of NF-κB,STAT-3,SMAD3 and AP-1 in acutely damaged liver

AIM: To investigate the role of genes and kinetics of specific transcription factors in liver regeneration, and to analyze the gene expression and the activity of some molecules crucially involved in hepatic regeneration. METHODS: USING gel-shift assay and RT-PCR, transcription factors, such as NF-κB, STAT-3, SMAD3 and AP-1, and gene expression of inducible nitric oxide synthase (iNOS), hepatocyte growth factor (HGF) and c-met were analyzed in an animal model of chemically induced hepatectomy. RESULTS: Gene expression of HGF and its receptor c-met peaked at 3 h and 24 h after acute CCl4 intoxi- cation. iNOS expression was only observed from 6 to 48 h. Transcriptional factor NF-κB had an early activation at 30 min after acute liver damage. STAT-3 peaked 3 h post- intoxication, while AP-1 displayed a peak of activation at 48 h. SMAD3 showed a high activity at all analyzed times. CONCLUSION: TNF-α and IL-6 play a central role in hepatic regeneration. These two molecules are responsible for triggering the cascade of events and switch-on of genes involved in cell proliferation, such as growth factors, kinases and cyclins which are direct participants of cell proliferation.

Expression of Livin in tissues of lung cancer and its correlation with the expression of caspase-3

Objective： To study the expressions of two isoforms of Livin in tissues of lung cancer and their relations to histological types and chemotherapy, and to study their correlations to the expression of caspase-3 as well. Methods： Expressions of Livin isoforms a, 13 and caspase-3 were detected by reverse transcription polymerase chain reaction （RT-PCR） assay in lung cancer tissues as well as in controls. Results： Livin isoforms a and ~ were expressed in 12 of 27, and 19 of 27 lung cancer tissues respectively, much more than those in lung para-cancereus [both were （0/6）] or benign disease lung tissues （0/12, 1/12; P 〈 0.01 and P 〈 0.01 ）. Moreover, they were detected in 7/14, 9/14 lung adenocarcinomas and 4/12, 9/12 squamocallular and large call carcinomas, respectively, and both showed expressions in one small cell carcinoma. The levels of these two isoforms in lung cancer were significantly higher than those in controls by Gel imaging system （P 〈 0.05 and P 〈 0.05）, the former was higher in adenocarcinoma than that in squamocellular carcinoma （P 〈 0.05）, while the latter was the same in both （P 〉 0.05）. Meanwhile, the levels of caspase-3 in lung cancer were significantly lower than those in controls, and it was suggested to be negatively associated with either each of two isoforms or their sum （P 〈 0.05, P 〈 0.01 and P 〈 0.01）. Two isoforms of Livin expression seemed to increase＇after chemotherapy but not related to clinical stages （P 〉 0.05）. Conclusion： Two isoforms of Livin are differently expressed in different histological types of lung cancer and may contribute to corresponding cancerous development; the levels of Livin are negatively associated with those of caspase-3, this may be due to the fact that Livin could resist against apoptosis; high expression of Livin seems to be related to chemotherapy but not clinical stages.

Influence of neurotrophin-3 on Bcl-2 and Bax expressions in spinal cord injury of rats

Objective： To study the protective mechanisms of neurotrophin-3 （NT-3） on the spinal cord injury. Methods：Totally 105 SD rats were randomly divided into 3 groups： control group, experimental group and sham operation group. Rats from the former 2 groups were inflicted to animal model of acute spinal cord injury according to Allen＇s （WD） by situating a thin plastic tube in the subarachnoid space below the injury level for perfusion. Rats in experimental group received 20 ul NT-3 （200 ng） from the tube at 0, 4, 8, 12, 24 h and 3, 7 d after injury, and those in control group got an equal volume of normal saline at the same time. The animals in sham operation group only received opening vertebral plate and tube was put in subarachnoid space. The rats were sacrificed at 4, 8, 12, 24 h and 3, 7, 14 d post injury （n=5）. The expression levels of Bcl-2 and Bax proteins in spinal cord of rats were detected by immunohistochemistry assay. Results： The level of Bax protein in control group significantly increased as compared with those in sham operation group, and the peak reached at 8 h after spinal cord injury. The Bcl-2 proteins were always weakly positive. The Bax proteins in NT-3 group significantly decreased but the Bcl-2 proteins obviously increased as compared with those in control group. Conclusion： NT-3 can protect spinal cord from injury in vivo. One of the mechanisms is that NT-3 can inhibit abnormal expression of Pax protein, and increase the expression of Bcl-2 protein, then inhibit apoptosis after spinal cord injury.

EXPRESSION OF HUMAN BETA-DEFENSIN 3 IN COS-7 CELL

To establish a cell line for stable expression of human beta-defensin 3 (hBD3). Methods Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofe-ctamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated. Results COS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity. Conclusion We successfully established a hBD3-expressing cell line.

EFFECTS OF PHYCOCYANIN ON EXPRESSION OF CytC mRNA AND CASPASE-3 mRNA AFTER FOCAL CEREBRAL ISCHEMIA/REPERFUSION IN RATS

Objective To study the effects of phycocyanin on the expression of Cytochrome C (CytC)genes and Caspase-3 genes after focal cerebral ischemia/reperfusion in rats. Methods A rat middle cerebral ar-tery occlusion (MCAO)/reperfusion model was produced using the intraluminal filament method. The rats were di-vided into three groups: sham operation group, model control group and phycocyanin group. After MCAO, the neu-robehavioral testing of all rats was made. The infarction area was evaluated with the method of 2,3,7-triphenyltet-razolium chloride (TTC) staining. The expression of CytC mRNA and Caspase-3 mRNA were determined by in situhybridization. Results In the sham operation group and the model control group, there was only a few CytC-positive cells were seen in the normal cerebral tissue. In the model control group, the upregulation of CytC mRNAbegan 6h after ischemia, reached a maximum at 12h (cortex) -24h (striatum) , then subsided gradually, but stillin high level. In the phycocyanin group, CytC-positive cells were also mainly in cortex and striatum, but the numberof the cells was significantly lower than the number of the model control group. The time-phase pattern of CytCmRNA in the phycocyanin group was similar to the pattern of the model control group. In the sham operation groupand the model control group, there was only a few Caspase-3-positive cells were seen in the normal cerebral tissue.In the model control group, the upregulation of Caspase-3 mRNA began 6h after ischemia, reached a maximum at24h and subsided at 48h, but still in high level. In the phycocyanin group, Caspase-3-positive cells were also mainlyin the penumbral area, but the number of the cells were significantly lower than the number of the model controlgroup. The time-phase pattern of Caspase-3 mRNA in the phycocyanin group was similar to the pattern of the modelcontrol group. Conclusion The over-expression of CytC mRNA and Caspase-3 mRNA might play a key role inischemic cerebral injury after MCAO. Phycocyanin could inhibit the over-expression of CytC mRNA and Caspase-3mRNA in the cerebral cortex, and might play an important role in the protection of ischemic neurons.

Treatment of genotype 2 and 3 chronic hepatitis C virus-infected patients

AIM： Before pegylated interferon alpha （IFN） was introduced for the therapy of chronic hepatitis C virus （HCV）-induced hepatitis, conventional thrice weekly IFN therapy was supplemented by ribavirin. Also, at that time, higher and more frequent doses of IFN were expected to be more effective than the standard regimen of 3 MU thrice weekly. As ribavirin significantly increases side effects and negatively influences the quality of life particularly in young patients, we started a prospective non-randomized study with a daily IFN-2a monotherapy as an initial treatment for chronic hepatitis C. METHODS： Forty-six consecutive chronic HCV-infected patients received 3 MU IFN-2a per day as an initial treatment. Patients with genotype 2 or 3 （n = 12） were treated for 24 wk, and patients with genotypes other than 2 or 3 （n = 34） for 48 wk. Treatment outcome was followed up for 48 wk after the end of treatment （EOT）. Virological response was defined as the absence of detectable serum HCV-RNA. Patients without virological response at 12 wk after the start of treatment received low-dose ribavirin （10 mg（kg·d）） additionally. RESULTS： During treatment, three genotype 3 patients were excluded from the study due to incompliance. The remaining patients （n = 9） infected with genotype 2 or 3 showed an initial virological response rate of 100%. Six patients （66.7%） were still found to be virus-free at the end of follow-up period. In these patients, initial virological response was evident already after 2 wk of treatment. In contrast, initial virological response occurred first after 4 wk of treatment in the three patients who relapsed （33.3%）. In comparison, patients infected with genotypes other than 2 or 3 （n = 34） showed an initial virological response rate of only 23.5% （n = 8）, and even in combination with ribavirin a sustained virological response （SVR） rate of only 11.8% （n = 4） could be achieved. CONCLUSION： In chronic HCV-infected patients with genotype 2 or 3, a SVR can be expected after 24 wk of daily dose IFN-2a treatment without ribavirin, if initial virological response develops early. This finding is worth to be confirmed in a prospective randomized study with pegylated IFN.

Cloning and Expression of a Profilin Gene from Rapeseed

Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA clone (designated Repro) encoding profilin gene was isolated from rapeseed ( Brassica napus L. cv. canadian Tween) using RT-PCR technique. Sequence analysis showed 82% similarity to Zea mays L. ZmPro3, 85% to Arabidopsis AthPRF1, 82% to Nicotiana tabacum L. NTPRO, 81% to Oryza sativa L. profilin A. A new full-length cDNA was obtained by 5'-RACE and 3'-RACE techniques. Sequence analysis showed that the size of full-length cDNA is 672 bp which contains a major open reading frame of 134 amino, acids, 5' and 3' untranslated regions and a long Poly (A) tail. Northern blot analysis showed that the profilin gene is a pollen and anther specific gene.

Cloning of Syntaxin Gene in Limonium sinense Kuntzet

[Objective] The aim was to clone Syntaxin genes in Limonium sinense Kuntze. [Method] Limonium sinense Kuntze leaves were used as materials and total RNA was extracted and transcribed reversely. Nested primers were designed based on EST sequences at 5’ region of Syntaxin, and cDNA obtained through reverse reaction was taken as the template. Sequences of Syntaxin gene at 3’ region were obtained through two rounds of PCR amplifications. [Result] DNA fragments （1 096 bp） were obtained. For LsSyntaxin, open reading frame （ORF） was 816 bp and the encoded amino acids were 271. The relative molecular weight of Syntaxin was 30 254.3 Da and isoelectric point in theory was 5.55. [Conclusion] Syntaxin genes from Limonium sinense Kuntze were cloned. The research laid foundation for the study on Syntaxin gene function in Limonium sinense Kuntze and salt-secreted process.

Hepatitis C virus nonstructural protein NS_3 and telomerase activity

OBJECTIVE: To study the effect of hepatitis C virus nonstructural protein NS(3) (HCV NS3) on telomerase activity and carcinogenesis. METHODS: Streptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS(3) protein in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' and pRcHCNS(3)-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay (ELISA) technology in the transfected and non-transfected NIH3T3 cells. RESULTS: HCV NS(3) protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' expressing HCV NS(3) C-terminal deleted protein or with plasmid pRcHCNS(3)-3' expressing HCV NS(3) N-terminal deleted protein. The positive signal of HCV NS(3) protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P

Altered gene expression reveals molecular mechanisms underlying oridonin-induced apoptosis of multiple myeloma LP-1 cells

Objective： To investigate the effect of oridonin on proliferation and invasion of human multiple myeloma LP-1 ceils and the underlying mechanism. Methods： LP-1 cells in culture medium in vitro were treated with oridonin at the different concentration Cell proliferation was measured by Microwave Theory and Techniques （MTT） assay and cell apoptotic rate was detected by flow cytometry. Morphology of cell apoptosis was observed by transmission electron microscope. Expressions of Bax, Bcl-2, Caspase-3, NFqcB as well as I-~B mRNA were detected by real-time PCR. Results： The MTT assays and flow cytometry revealed that oridonin could inhibit the growth of LP-1 cells and cause apoptosis significantly; the suppression was both in time- and dose-dependent manner. Marked morphological changes of cell apoptosis were found under a transmission electron microscope after the cells were treated with oridonin at 25 ~rnol/L for 24 h. Along with the apoptotic process, Bcl-2, Caspase-3,NF-r,.B gene expressions were down-regulated （P〈0.05）. On the contrast, the Bax and I-~zB gene expressions were up-regulated （P〈0.05）. Conclusion： Oridonin could inhibit the proliferation of LP-1 cells via inducing apoptosis. We concluded that oridonin induces apoptosis in LP-1 cells via activation of caspase-3 as well as down-regulation of Bcl-2 and up-regulation of Bax expression. The results suggested that oridonin could induce apoptosis of LP-1 cells through mitochondria- and caspase3-dependent pathways. Meanwhile, the inhibition of NF-r,_B and the activation of I-~B indicate pro-apoptotic stimuli. In one word, oridonin might be an important potential anti-myeloma reagent.

Expression of the apoptosis-related proteins caspase-3 and NF-κB in the hippocampus of Tg2576 mice

Objective To investigate the relations between neuroapoptosis and the onset and development of Alzheimer’s disease （AD）, especially the role of NF-κB in the regulation of neuroapoptosis. Methods Caspase-3 and NF-κB （p50） expressions in the CA3 region of the hippocampus in APPswe Tg2576 transgenic mice were studied from postnatal day 0-180, using Nissl staining, immunohistochemistry and RT-PCR methods. Results Both neuronal apoptosis and NF-κB activity decreased gradually with the increase of age in wild type and Tg2576 mice. However, the number of caspase-3-positive or NF- κB-positive pyramidal cells in Tg2576 mice was greater than that in age-matched wild type mice, with significant differences after postnatal day 14 （P 0.01 or P 0.05）. Linear regression analyses of caspase-3 and NF-κB expression demonstrated a correlation between neuroapoptosis and activity of NF-κB. Conclusion The process of neuroapoptosis is consistent with the onset and development of AD. Furthermore, the observed correlation between neuroapoptosis and NF-κB activity suggests a role of NF-κB in hippocampal neuroapoptosis.