Our aim in this study was to determine differentially expressed genes of CD34 + cell from bone marrow and G CSF mobilized peripheral blood. CD34 + cells isolated from bone marrow and G CSF mobilized peripheral blood of the same donor were identified for differentially expressed genes in CD34 + cells using the technique of suppression subtractive hybridization. Eleven genes were expressed with higher levels in CD34 + cells from mobilized peripheral blood, as compared with data of GenBank. These genes included nuclear proteins, transcriptional regulatory molecules, zinc finger proteins and interferon induced molecules. These results demonstrate that there are some differences between the two groups of CD34 + cells. Further study would give us more extensive understanding about mobilization and homing of hematopoietic stem cells.

Objective: To investigate the expression of MDR1 and GFP in the human hematopoietic cells mediated by adeno-associated virus. Methods: The GFP gene was transferred into the human hematopoietic cells by AAV vectors and created strong visible fluorescence by purely molecular biological means. Using adeno-associated virus vectors, we have transferred human mdr-1 gene into human hematopoietic cells and investigated the drug resistence of human hematopoietic cells modified with mdr-1 gene. PCR analysis confirmed that mdrl cDNA had been successfully transferred into the human hematopoietic cells. An assay of MTT proved that the human hematopoietic cells modified by mdrl gene had resistance to colchicine. Results: It was about 30% of the hematopoietic cells that expressed the green fluorescent proteins. The resistance of hematopoietic cells was increased parently when the cells were infected by the crude virus stocks. Conclusion: It is conducted that the AAV vector could successfully transfer the foreign gene into the human hematopoietic cells. The cells modified with mdrl gene have increased the resistance to drugs.

Breast cancer is the leading cause of malignancy-related mortality in women worldwide. The more accurate prediction of lymph node metastasis and evaluation of personalized prognosis of breast cancer patients could provide evidence and reference for individualized comprehensive treatment and clinical decision-making. Nomogram is statistical calculation model developed to generate individualized prediction of a certain clinical event through the factors associated with it. Currently breast cancer related nomogram models is most commonly used in the prediction of non-sentinel lymph node status in patients with sentinel lymph node-positive breast cancer, sentinel lymph node metastasis in clinical node-negative breast cancer and prognosis evaluation of breast cancer. This article reviewed the recent advances in breast cancer related nomograms according to the above mentioned three aspects, and evaluated respectively the predictive factors, accuracy, characteristics and clinical application potential.

Objective:To investigate association of the new polymorphism G352A in the dopaminetransporter gene(DAT1)exon 15 with attention deficit hyperactivity disorder(ADHD)in Han Chinesechildren.Methods:The new mutant polymorphism G352A in the dopamine transporter gene(DAT1)ex-on 15 was found by the fluorescently-labeled dye-terminators assay.The study samples were comprised of337 ADHD children,207 unrelated controls and 201 integrated ADHD trios(included proband and bio-logical parents).Associations of polymorphisms with ADHD and its subtypes were examined by:(i)comparing cases and controls;and(ii)using family-based association study in transmission-disequilibri-um test(TDT).Results:The allele frequencies at the DAT1 G352A locus in the control samples were79.5% for 352G and 20.5% for 352A respectively.Association studies revealed no association betweenG352A in exon 15 of DAT1 and ADHD.But after a stratification by gender,there was possible associationbetween G352A and ADHD girls:the 352G allele had a tendency to be preferentially transmitted toADHD girls.Conclusion:There is no association between G352A,the new polymorphism,in exon 15 ofDAT1 and ADHD.The 352G allele has a tendency to be preferentially transmitted to ADHD girls,but thefindings require replication before drawing a definitive conclusion.

AIM To establish a model of vascular intima hyperplasia for study the restenosis after angioplasty. METHODS Carotid artery of rat was exposed under general anesthesia. Two pieces of steel slides(13 mm?5 mm?1 mm) were set on and under the carotid artery separately. Then the slides were squeezed by two surgical forceps paralleling to the artery for 25 min. Histomorphological study was performed at the second hour or on the fourteenth day after operation. RESULTS At the second hour after operation, the loss of endothelial integrity and platelet deposition were seen under the scanning electromicroscopy. Under the light microscopy, there were infiltration of inflammatory cells in vessel wall andthrombus formation in the vessel cavity. At the fourteenth day after operation, PCNA positive cells in vessel wall of squeezed artery were significantly higher, the ratios of intima, medium and intima areas of squeezed artery were significantly increased and the ratio of cavity area of squeezed artery was significantly decreased compared with those of uninjured artery.The vascular intima hyperplasia and PCNA positive cells in squeezed arterial wall were inhibited by oral administration of heparinoid and aspirin. CONCLUSION With squeezing carotid artery in rats the vascular intima hyperplasia as restenosis after angioplasty was established.

Objective:To explore the expression of N-cadherin and its correlations with the clinicopathological features of human ovarian carcinoma.Methods:The expression of N-cadherin in 281 cases of ovarian carcinoma tissues were determined by immunohistochemical method.The correlations of N-cadherin expression with the clinicopathological features of human ovarian carcinoma were analyzed.Results:There was higher expression of N-cadherin in the metastatic lesions than its paired primary lesions (P =0.018).The expression level of N-cadherin in ovarian carcinoma was correlated with the FIGO stage (P =0.034),histological type (P ＜0.001) and tumor grade (P =0.004).Conclusions:High expression of N-cadherin might positively correlate with the invasion and migration ability of ovarian carcinoma cells,which was more common in the the advanced (FIGO Ⅱ-Ⅳ) ovarian carcinoma,high-grade serous carcinoma,and high grade ovarian carcinoma.N-cadherin might be useful in estimating the biological behavior of human ovarian carcinoma.

To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.
Adipose Tissue, Brown ; drug effects ; physiology ; Animals ; Capsaicin ; analogs & derivatives ; pharmacology ; Energy Metabolism ; Ganglia, Spinal ; cytology ; HEK293 Cells ; Humans ; Mice ; Neurons ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; TRPV Cation Channels ; physiology ; Temperature ; Thermogenesis

The aim of this study was to construct recombinant mDHFR-GFP/AAV vector containing mutated dihydrofolate reductase (mDHFR) and green fluorescent protein (GFP) fusion genes and its expression in NIH3T3 cells, to investigate the resistance of the cells to methotrexate. Amplified cDNA of mDHFR and GFP segmented from their plasmid separately were linked by PCR with the aminoacetic acid linker. The fusion gene was inserted into T vector, and after enzyme cutting the fusion gene fragment was inserted into AAV vector, then packaging the vector into recombined AAV and infected NIH3T3 cells. Expression of gene fusion was observed by PCR, fluorescent microscopy and flow cytometry. mDHFR and GFP cDNA were found in NIH3T3 genomic DNA, the GFP expression rate was about 25%, and resistance of the transferred cells to MTX was increased markedly. The results showed that AAV vector can transfer mDHFR and GFP fusion gene into NIH3T3 cells and increase resistance to MTX in gene modified cells. This data provided a basis for application of mDHFR and AAV vector in gene therapy.
3T3 Cells ; Adenoviridae ; genetics ; Alcohol Oxidoreductases ; genetics ; Animals ; Cell Survival ; DNA Restriction Enzymes ; metabolism ; DNA, Complementary ; genetics ; metabolism ; Flow Cytometry ; Gene Expression ; Genetic Vectors ; genetics ; Mice ; Microscopy, Fluorescence ; Mutation ; Recombinant Fusion Proteins ; genetics ; metabolism

OBJECTIVEThis study was designed to examine the in vitro effects of fenvalerate on steroid production and steroidogenic enzymes mRNA expression level in rat granulosa cells.

METHODSUsing primary cultured rat granulosa cells (rGCs) as model, fenvalerate of various concentrations (0, 1, 5, 25, 125, 625 micromol/L) was added to the medium for 24 h. In some cases, optimal concentrations of 22(R)-hydroxycholesterol (25 micromol/L), Follicle stimulating hormone (FSH, 2 mg/L), or 8-Bromo-cAMP (1 mmol/L) were provided. Concentrations of 17 beta-estradiol(E2) and progesterone (P4) in the medium from the same culture wells were measured by RIA and the steroidogenic enzyme mRNA level was quantified by semi-quantitative RT-PCR.

RESULTSFenvalerate decreased both P4 and E2 production in a dose-dependent manner while it could significantly stimulate rGCs proliferation. This inhibition was stronger in the presence of FSH. Furthermore, it could not be reversed by 22(R)-hydroxycholesterol or 8-Bromo-cAMP. RT-PCR revealed that fenvalerate had no significant effect on 3 beta-HSD, but could increase the P450scc mRNA level. In addition, 17 beta-HSD mRNA level was dramatically reduced with the increase of fenvalerate dose after 24 h treatment.

CONCLUSIONFenvalerate inhibits both P4 and E2 production in rGCs. These results support the view that fenvalerate is considered as a kind of endocrine-disrupting chemicals. The mechanism of its disruption may involve the effects on steroidogenesis signaling cascades and/or steroidogenic enzyme's activity.

3-Hydroxysteroid Dehydrogenases ; analysis ; metabolism ; 8-Bromo Cyclic Adenosine Monophosphate ; pharmacology ; Animals ; Base Sequence ; Cells, Cultured ; Dose-Response Relationship, Drug ; Estradiol ; analysis ; metabolism ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Hydroxycholesterols ; pharmacology ; Nitriles ; pharmacology ; Progesterone ; analysis ; metabolism ; Pyrethrins ; pharmacology ; RNA, Messenger ; analysis ; metabolism ; Rats ; Steroids ; metabolism