乳腺照相辅以导丝定位活检对诊断乳腺原位癌的意义

目的 探讨乳腺X线钼靶照相辅以细针穿刺导丝定位活检技术对提高原位癌诊断率的意义。方法 收集 1997-2 0 0 0年病理确诊的乳腺癌 679例 ;以及近半年来手术或活检病理证实的乳腺癌 74例 ,术前 5 1例行乳腺钼靶照相 ,其中 4例行定位活检。比较两个时期乳腺原位癌的发现率。结果 1997-2 0 0 0年 679例乳癌患者原位癌 18例 ,占乳癌 2 .7%。近半年来 74例乳癌患者中 ,原位癌 4例 ,发现率为 5 .4%。比 1997-2 0 0 0年明显增高。结论 乳腺钼靶X线照相及穿刺定位活检技术可以提高原位癌的诊断率。

Effects of Fungal Elicitors on Cell Growth and Artemisinin Accumulation in Hairy Root Cultures of Artemisia annua

The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr.) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae , the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.

Signal Transduction from Water Stress Perception to ABA Accumulation

To cope with unpredictably environmental perturbations and sometimes stresses, plants have evolved with some mechanisms so that these developing stresses can be sensitively perceived and the physiology can be rapidly regulated. Such perception and regulation can be a kind of feed_forward mechanism and may involve many biochemical and physiological processes and/or the expression of many genes. Although many dehydration_responsive genes have been identified, much fewer of their functions have been known. Such stress_ induced responses should include the initial perception of the dehydration signal, then the complicated signal transduction and cellular transmission until to the final gene activation or expression. As an important plant stress hormone abscisic acid (ABA) mediates many such responses. We believe that starting from the initial perception of dehydration to the gene expression leading to the stress_induced ABA biosynthesis is the most important stress signal transduction pathway among all the plant responses to stresses. Identification of the genes involved and understanding their roles during stress perception and physiological regulation shall be the most important and interesting research field in the coming years.

The Mediation of Defense Responses of Ginseng Cells to an Elicitor from Cell Walls of Colletotrichum lagerarium by Plasma Membrane NAD(P)H Oxidases

NAD(P)H oxidases were detected in suspension cultured cells of ginseng (Panax ginseng C. A. Meyer). The activities of these enzymes were induced by an elicitor (Cle) extracted from cell walls of Col-letotrichum lagerarium. In addition, Cle induced an oxidative burst and enhanced the synthesis of saponin, activity of phenylalanine ammonialyase (PAL) , accumulation of chalcone synthase (CHS) and the transcription of a hydroxyproline-rich glycoprotein gene ( hrgp ) . Pre-treatments with DPI and quinacrine (two inhibitors of mammalian neutrophil plasma membrane NADPH oxidase) for 30 min prior to Cle addition blocked the NAD(P)H oxidase activity induced by Cle. These inhibitors also inhibited the release of H2C2, the synthesis of saponin, PAL activity and CHS accumulation. Our data revealed homology between plasma membrane NAD(P)H oxidases of mammalian neutrophil cells and ginseng suspension cells. They also indicated that deactivated NAD(P)H oxidases catalysed the release of H2O2 and that H2O2 was functioning as a second messenger stimulating PAL activity, saponin synthesis and hrgp transcription. Elevations of Ca2 + and protein phos-phorylation/dephosphorylation were required for this defense process. We propose that NAD(P)H oxidases mediate the processes of Cle-induced defense responses in ginseng suspensions, and postulate the existence of a signalling cascade including extracellular Cle stimulation, activation of plasma membrane NAD(P)H oxidases, release of H2O2, and the intracellular responses of metabolism and gene transcription in ginseng suspension cells.

浅教之中有真意——肖培东课堂教学艺术例谈

探讨肖培东老师的浅教艺术。以具体课例说明浅教就是以培养学生正确理解和运用祖国语言文字的能力为核心,老老实实地把学生引领到读、写、听、说的语文实践中。

Culture of Porcine Peripheral Blood Monocyte-derived Dendritic Cells in vitro

[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.

Involvement of Calcium dependent Protein Kinases in ABA regulation of Stomatal Movement

Patch clamp techniques were employed to investigate if calcium dependent protein kinases (CDPKs) be involved in the signal transduction pathways of stomatal movement regulation by the phytohormone abscisic acid (ABA) in Vicia faba. Stomatal opening was completely inhibited by external application of 1 μmol/L ABA, and such ABA inhibition was significantly reversed by the addition of CDPK inhibitor trifluoperazine (TFP). The inward whole cell K + currents were inhibited by 60% in the presence of 1 μmol/L intracellular ABA, and this inhibition was completely abolished by the addition of CDPK competitive substrate histone Ⅲ S. The results suggest that CDPKs may be involved in the signal transduction cascades of ABA regulated stomatal movements.

Key factors for causing poplar Ice Nucleation Active bacterial canker and its control techniques

The isolation, culture and the active determination of poplar ice nucleation active (INA) bacteria and the inoculation tests in laboratory and field were conducted, and the varieties, distribution and number of poplar INA bacteria and its pathoge-nicity and freezing injury property were determined. The study results showed that the INA bacteria widely spread on poplar in Northeast China and caused the frozen injury for poplar under the frost condition in Spring or Autumn, which was the key factor to induce INA bacterial canker. Through evaluation and investigation of different poplar varieties and inoculation tests, fine dis-ease-resistant varieties and strains of poplar suitable for Northeast China were selected. Further tests for strong seedling showed that burying cuttings in sand and covering with plastic film could effectively avoid the frostbite, frozen and drought damage, reduce INA bacteria infection, and promote poplar growth. INA bacterial canker was detected early by highly special-ized antiserums of INA bacteria and the agglutinated test of ring-shaped boundary surface. The inducers such as streptomycin, phenylmercuric acetae, salicylic acid and heat-killed bacteria to immerse cuttings, have obvious induced disease-resistant effect. Before poplar sprouted in early spring, through spraying the solution of frostbite agent, the control effect also was obvious.

Induced Expression of Cytochrome P450 CYP305 B1V1 Gene in Different Tissues of Wild Mulberry Silkworm(Bombyx mandarina)

[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA sequence of CYP305 B1 V1 Gene published in GenBank for wild mulberry silkworm, one pair of primers was designed, and the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm treated by NaF, rutin, cypermethrin and ecdysone was also analyzed by the semi - quantitative RT - PCR. Furthermore, homology comparison and phylogenetic analysis for amino acid sequences of this gene were studied. [ Result] Rutin, cypermethrin and NaF had effects on the expression of P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm, while ecdysone had no significant effect. Homology comparison for amino acids indicated that the amino acid sequence of this gene was the most similar to that of CYP305 B1 gene in Bombyx mori with 100% amino acid identity, and highly similar to those of Tribolium casmneum CYP305A1, Apis mellifera CYP305A1, Drosophi- la melanogaster CYP305A1, Anopheles gambiae CYP305A2and Culex pipiens quinquefasciatus CYP2LI. [ Conclusion] CYP305 B1 V1 Gene of wild mulberry silkworm is likely to mainly take part in the metabolism of exogenous compounds, which is of great significance for revealing the function of cytochrome P450 and the metabolic mechanism of different drugs.

Anti-CD132 Monoclonal Antibodies Inducing T Cells Apoptosis after Alloantigen Stimulation and Its Possible Clinical Applications

Aim To investigate the mechanism of anti-CD132 monoclonal antibodies (mAbs)inhibiting T cells proliferation in vitro, and their potential values for clinical use. MethodsBALB/c and C57BL/6 mice splenocytes were harvested for two-ways mixed lymphocyte culture (MLC).Anti-CD132 mAbs (final concentration 100 mg·L^(-1)) were added in MLC on day 0 (group 1) or day 3(group 2). Fluorescence activated cell sorting (FACS) was used to measure the proliferation(carboxy-fluorescein dia cetate, succinimidyl ester, CFSE), apoptosis of T cells (PE-CD3,FTTC-annexin-v), and cell cycle (pro-pidium iodide stain) . The expression of survivin in T cellswas detected by immunochemical stai-ning. Re-sults Multi-generation CFSE-labeled splenocytes werefound dividing and their fluorescent strength decreased in MLC. There was no noticeable change influorescent intensity in group 1 and group 2. On day 3, apoptosis induced by anti-CD132 mAbs wasdetected in part of T cells, but was not detected in the former two days in group 1. In group 2, thenumber of cells in M phase (activated T cells) decreased and apoptot-ic cells increased on day 4.The phenomena were not observed in control group (P < 0.01). Expression of survivin in T cells wasdetected in control group but not in groups 1 and 2. Conclusion Blockade of CD132 signaling pathwayinhibits T cell proliferation in vitro by means of inducing activated alloreactive T cell apoptosisbut not the resting T cells. Anti-CD132 mAbs may be candidates for clinical applications.

Involvement of cAMP in ABA Signal Transduction in Tobacco Suspension Cells

Abscisic acid (ABA) plays an important role in plant growth and developmental processes. Although some ABA signal molecules, such as cADPR, Ca2+, etc., have been reported, there. was no evidence proving the involvement of cAMP in A-B-A, signal transduction. In this present study, the constructed gene ( rd29A-GUS) was transformed into Nicotiana tabacum, and calli was induced from the transgenic plant. The suspension cells obtained from the callus grew well and uniformly. Treatment of the suspension cells with ABA led to an increase in GUS activity, indicating that these transgenic suspension cells are useful for the study of ABA signaling. Addition of nicotinamide (cADPR inhibitor) or U-73122 (phospholiphase C inhibitor) could only partially inhibit the increase of GUS activity elicited by ABA. The inhibitory effect of nicotinamide was enhanced by application of K252a (inhibitor of protein kinase). Treatment of the suspension cells with 8-Br-cAMP, a membrane-permeable analogue of cAMP, could partially replace the effect of ABA. Furthermore, intracellular addition of IBMX (phosphodiesterase inhibitor) mimicked die effect of exogenous cAMP on the deduction of expression of rd29A promoter. These results suggested that cAMP was an important messenger in ABA signal transduction in tobacco suspension cell.

Effect of Activation-induced Cell Death of Peripheral Blood Mononuclear cells in Patients with Condyloma Acuminatum

Objective: To investigate the effect of activation-induced cell death (AICD) on cellular immune function in the condyloma acuminatum(CA). Methods: Peripheral blood mononuclear cells (PBMC) were isolated from normal healthy individuals (control group) and patients with CA. PBMC were cultured with PHA-P for 48h in vitro. Apoptosis of the PBMC was detected by flow cytometry. Supernatant cytokines (IL-2 and IL-10) were assayed by ELISA. Results: The rate of PBMC apoptosis in both CA group and control group in fresh PBMC was very low and similar in both groups(P>0.05). The rate of PBMC apoptosis within the CA group was noticeably increased compared to that of the control (P<0.001)af-ter PBMC were cultured for 48h. The level of IL-2 was significantly lower in the CA group than in the control group (P<0.001), The level of IL-10 was significantly higher in the CA group compared to thecontrol group(P<0.001). Conclusion: Study results indicate that AICD may affect cellular mediated immune function and play an important role in the pathogenesis of CA.

Desferoxamine preconditioning protects against cerebral ischemia in rats by inducing expressions of hypoxia inducible factor 1α and erythropoietin

Objective To investigate whether desferoxamine （DFO） preconditioning can induce tolerance against cerebral ischemia and its effect on the expression of hypoxia inducible factor 1 α （HIF- 1α） and erythropoietin （EPO） in vivo and in vitro. Methods Rat model of cerebral ischemia was established by middle cerebral artery occlusion with or without DFO administration. Infarct size was examined by TTC staining, and the neurological severity score was evaluated according to published method. Cortical neurons were cultured under ischemia stress which was mimicked by oxygen-glucose deprivation （OGD）, and the neuron damage was assessed by MTT assay. Immunofluorescent staining was employed to detect the expressions of HIF-1 and EPO. Results The protective effect induced by DFO （decreasing the infarction volume and ameliorating the neurological function） appeared at 2 d after administration ofDFO （post-DFO）, lasted until 7 d and disappeared at 14 d （P 〈 0.05）; the most effective action was observed at 3 d post-DFO. DFO induced tolerance of cultured neurons against OGD： neuronal viability was increased 23%, 34%, 40%, 48% and 56% at 8 h, 12 h, 24 h, 36 h, and 48 h, respectively, post-DFO （P 〈 0.05）. Immunofluorescent staining found that HIF-1 α and EPO were upregulated in the neurons of rat brain at 3 d and 7 d post-DFO; increase of HIF-1 α and EPO appeared in cultured cortex neurons at 36 h and 48 h post-DFO. Conclusion DFO induced tolerance against focal cerebral ischemia in rats, and exerted protective effect on OGD cultured cortical neurons. DFO significant induced the expression of HIF- 1 α and EPO both in vivo and in vitro. DFO preconditioning can protect against cerebral ischemia, which may be associated with the synthesis of HIF- 1 α and EPO.

Expression of Hypoxia Inducible Factor lα and Its Significance in Non-small Cell Lung Cancer

Objective： To explore the expression level and its clinical significance of hypoxia inducible factor 1α （HIF-1α ） in non-small lung cancer. Methods： The expression of HIF-1α was detected in 68 human non-small lung cancer samples by immunohistochemistry. Results： （1） Thirty-nine （57.35%） out of the 68 human non-small lung cancer samples was positive for HIF-1α ; （2） The positive rate of HIF-1α in adenocarcinoma and squamous carcinoma was 54.76% （23/42） and 61.54% （16/26） respectively. No significant difference was found between adenocarcinoma and squamous carcinoma of non-small lung cancer in the expression of HIF-1α （P〉0.05）. The positive rate of HIF-1α in middle-high differentiation was 74.28% （26/35）, significantly higher than in low differentiation （39.39%, 13/33） （P〈0.05）; （3） The positive expression of HIF-1α was not correlated to the sexes, ages, tumor stage and lymph node status. Conclusion： The expression of HIF-1α is higher in non-small lung cancer and is correlated to differentiation.

Polymethylenebis [acetamides] Analogues.Synthesis and Differentiation-Inducing Activity on HL-60 Cells

报导了一系列多亚甲基双［酰胺］类化合物的合成，由－摩尔多亚甲基二甲酰氯分别和二摩尔２－氨基噻唑啉及５－氨基－１－甲基吡啶酮反应制得。测定了其体外对ＨＬ－６０人早幼粒白血病细胞的分化诱导活性，初步结果表明：Ｎ，Ｎ｀－双吡啶酮基六二甲酰胺和Ｎ，Ｎ｀－双噻唑啉基八亚甲基二甲酰胺分别在０．１ｍｍｏｌ／Ｌ和０．５ｍｍｏｌ／Ｌ浓度时，诱导分化百分率可达６０％。此浓度下细胞存活率分别为２６％及２２％，其有效诱导浓度比ＨＭＢＡ低十倍。

Observation on Apoptosis of Erythroid Cells Induced by Dexamethasone

Using splenic cells of BALB/c mice infected with Friend Leukemia Virus (FVA) as a model of erythroid cells, we investigated the action of dexamethasone (Dex) to induce apoptosis of the cells in the presence of erythropoietin (EPO)——an apoptotic inhibitor in developing erythroid cells. The result indicated that after treatment with a certain range of Dex concentrations and prolonged incubation, the cells were characterized by the occurrence of DNA ladders which appeared on agarose electrophoresis. Transmission electron microscopy showed that karyopyknosis, chromatin condensation, dilatation of the perinuclear space, karyorrhexis, cytoplasmic vacuolization and cell fragmentation appeared in the cells depending on the dose of Dex and the treatment time. These results suggest that Dex could induce apoptosis in the developing erythroid cells as in other cells so far reported.

Study on the Equivalent Conformation of Retinoids

N-(4-Carboxy-phenyl)-3,5-di-t-butyl-4-hydroxy-benzamide (2) possesses structural prerequisite for cell differentiation inducing activity, which constitutes the therapeutic basis of all trans retinoic acid (ATRA) and analogues for the treatment of cancer and dermatosis. In addition to the similarity of the disposition of functional groups with ATRA, 2 shows a conformational equivalence to ATRA in terms of molecular shape, size, as well as the spatial arrangement of functional groups. However, the N methylated compound (3) is devoid of the activity. It owes the biological behavior to the conformational difference, because of the steric interference between N methyl group and the hydrogen atom of a phenyl ring. X ray crystallography, UV, and NMR were performed to investigate the difference.

PGC-la induces apoptosis in human epithelial ovarian cancer cells through a PPARy-dependent pathway

Peroxisome proliferator-activated receptor gamma （PPAR）,） coactivator-1 alpha （PGC-1α） coactivates multiple transcription factors and regulates several metabolic processes. The current study investigated the role of PGC-1α in the induction of apoptosis in human epithelial ovarian cancer cells. The PGC-1α mRNA level between human ovaries and human ovarian epithelial tumors was examined by quantitative RT-PCR. Less PGC- 1α expression was found in the surface epithelium of malignant tumors compared with normal ovaries. Overexpression of PGC-1α in human epithelial ovarian cancer cell line Ho-8910 induced cell apoptosis through the coordinated regulation of Bcl-2 and Bax expression. Microarray analyses confirmed that PGC-1α dramatically affected the apoptosis-related genes in Ho-8910 cells. Mitochondrial functional assay showed that the induction of apoptosis was through the terminal stage by the release of cytochrome c. Furthermore, PG-C- 1 α-induced apoptosis was partially, but not completely, blocked by PPAR）, antagonist （GW9662）, and suppression of PPAR）, expression by siRNA also inhibited PGC-1α-induced apoptosis in Ho-8910 cells. These data suggested that PGC-1α exerted its effect through a PPARγ-dependent pathway. Our findings indicated that PGC-1α was involved in the apoptotic signal transduction pathways and downregulation of PGC-1α may be a key point in promoting epithelial ovarian cancer growth and progression.

Genetic Correction and Hepatic Differentiation of Hemophilia B-specific Human Induced Pluripotent Stem Cells

Objective To genetically correct a disease-causing point mutation in human induced pluripotent stem cells （iPSCs） derived from a hemophilia B patient. Methods First, the disease-causing mutation was detected by sequencing the encoding area of human coagulation factor IX （F IX） gene. Genomic DNA was extracted from the iPSCs, and the primers were designed to amplify the eight exons of F IX. Next, the point mutation in those iPSCs was genetically corrected using CRISPR/Cas9 technology in the presence of a 129-nucleotide homologous repair template that contained two synonymous mutations. Then, top 8 potential off-target sites were subsequently analyzed using Sanger sequencing. Finally, the corrected clones were differentiated into hepatocyte-like cells, and the secretion of F IX was validated by immunocytochemistry and ELISA assay.Results The cell line bore a missense mutation in the 6th coding exon （c.676 C〉T） of F IX gene. Correction of the point mutation was achieved via CRISPR/Cas9 technology in situ with a high efficacy at about 22% （10/45） and no off-target effects detected in the corrected iPSC clones. F IX secretion, which was further visualized by immunocytochemistry and quantified by ELISA in vitro, reached about 6 ng/ml on day 21 of differentiation procedure. Conclusions Mutations in human disease-specific iPSCs could be precisely corrected by CRISPR/Cas9 technology, and corrected cells still maintained hepatic differentiation capability. Our findings might throw a light on iPSC-based personalized therapies in the clinical application, especially for hemophilia B.